BCL9 peptides and variants thereof

Abstract

To provide BCL9 peptides, in particular variants of the HD2 domain within BCL9 peptides.SOLUTION: Disclosed here are polypeptides derived from the HD2 domain of human B-cell CLL / lymphoma 9 (BCL9) protein and variants thereof, as well as their use in the diagnosis, prevention, and / or treatment of a disease or disorder. Also disclosed are methods of generating such polypeptides and variants thereof.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Priority claims This application claims priority to U.S. Patent Application Serial No. 62 / 583,820, filed November 9, 2017, the entire contents of which are incorporated herein by reference.

[0002] The present invention relates to BCL9 peptides, and in particular to variants of the HD2 domain within BCL9 peptides. [Background technology]

[0003] β-catenin is a multifunctional protein crucial for cellular homeostasis and processes such as embryogenesis, epithelial cell proliferation, and organ regeneration. However, aberrant β-catenin signaling can lead to altered transcriptional activation, which can enable tumor growth and development. β-catenin is normally phosphorylated and targeted for degradation by the axin complex; however, under stimulation of the Wnt signaling pathway, unphosphorylated β-catenin can accumulate. Under conditions in which the Wnt signaling pathway is activated, β-catenin binds to lymphoid enhancer factor / T cell factor (LEF / TCF), translocates to the nucleus, and stimulates the transcription of Wnt target genes, such as c-myc and CD44, which are involved in tumorigenesis (see Clevers and Nusse, Cell 149:1192-1205 (2012)). Summary of the Invention

[0004] In one general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: [Table 1] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; The polypeptide has a length of 6 to 30 amino acids.

[0005] In some embodiments, Xaa7 is CBA.

[0006] In some embodiments, Xaa7 is selected from Cha and Cpa.

[0007] In some embodiments, Xaa7 is selected from α-methyl L, DCha, N-methyl Cha, and allyl Gly.

[0008] In some embodiments, Xaa7 is selected from AC4C, A6C, Aze, Phe(4-Cl), (β-tBu-Ala), and Tle.

[0009] In some embodiments, Xaa7 is selected from Phe(4-Cl), 4-FPh, 3,4-diClPh, and Cha.

[0010] In some embodiments, Xaa4Xaa8Xaa5 comprises IQR.

[0011] In some embodiments, Xaa4Xaa8Xaa5 comprises I(N-methylQ)R.

[0012] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(N-methylR).

[0013] In some embodiments, Xaa4Xaa8Xaa5 comprises (CBA)QR.

[0014] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(homo R).

[0015] In some embodiments, Xaa4Xaa8Xaa5 comprises (N-methylI)QR.

[0016] In some embodiments, Xaa4Xaa8Xaa5 comprises IQQ, IQE, IQ(NMeArg), (Nle)QR, IQ(Nar), or IQC.

[0017] In some embodiments, the polypeptide comprises: [Table 2] During the ceremony, Xaa1 and Xaa2 are each independently selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa9 is selected from Q, E, N-methyl Q, N-MeGln, and peptoid Q; Xaa 10 is selected from T, N-methyl T, and DThr; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit.

[0018] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains LQTLR.

[0019] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains the L(N-methylQ)TLR.

[0020] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa11 contains L(N-methylQ)TL(homoR).

[0021] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes L(NMeGln)T(NMeLeu)R.

[0022] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes L(N-methyl Q)TL(N-methyl R).

[0023] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes LN-methylQTLR, LETLR, (CBA)QTLR, (CBA)(N-methylQ)TLR, LQT(CBA)R, L(N-methylQ)(N-methylT)LR, L(N-methylQ)T(Cha)R, L(N-methylQ)T(α-methylL)R, or L(N-methylQ)(DThr)LR.

[0024] In some embodiments, the polypeptide comprises: [Table 3] During the ceremony, Xaa 12 is selected from H, N-MeHis, Cys, N-MeCys, homoHis, and NHis; Xaa 13 is selected from R, N-methylR, homoArg, Cit, Nar, and Phe(4-guanidino); Xaa 14 is selected from E, Q, N-methyl E, N-methyl Q, N-methyl D, and NMeGln; Xaa 15 is selected from R, homoR, and N-methylR.

[0025] In some embodiments, Xaa 12 Xaa 13 Xaa14 Xaa 15 includes HRER.

[0026] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 includes HRQR.

[0027] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 include HR(N-methylE)R, HR(N-methylE)R, HR(N-methylQ)R, HR(N-methylD)R, H(N-methylR)QR, HRQ(homoR), HRQ(N-methylR), H(homoArg)QR, HRQ(NMeArg), HR(NMeGln)R, (N-MeHis)RQR, (Cys)RQR, (NMeCys)RQR, (homoHis)RQR, (NHis)RQR, H(Cit)(N-methylQ)R, H(Nar)(N-methylQ)R, or H(4-guanidino-Phe)(N-methylQ)R.

[0028] In some embodiments, the N-terminus of the polypeptide is modified with a moiety selected from acetyl, propionyl, hexanoyl, 3-phenylpropanoyl, 2-cyclohexylacetyl, diphenylacetyl, 3,5-dihydroxybenzoic acid, 4-(trifluoromethyl)benzoic acid, 5-phenylvaleric acid, 4-biphenylacetic acid, dimethyl, HOCH2CH2CO-, and palmitoyl-PEG4.

[0029] In some embodiments, the C-terminus of the polypeptide is modified with a moiety selected from NH, (β-Ala)(β-Ala), (β-Ala)(β-Ala)NH, GRKKRRQRRRPQK(PEG-palmitoyl)NH, K(PEG-palmitoyl)NH, GRKKRRQRRRPQNH, and 1-(2-aminoethyl)-4-methylpiperazine.

[0030] In some embodiments, the C-terminus of the polypeptide is modified at NH2.

[0031] In some embodiments, the C-terminus of the polypeptide is modified with (β-Ala)(β-Ala).

[0032] In some embodiments, the N-terminus of the polypeptide is modified with acetyl and the C-terminus of the polypeptide is modified with NH2.

[0033] In some embodiments, the polypeptide is selected from: [Table 4] TIFF2025122018000006.tif231161TIFF2025122018000007.tif234160TIFF2025122018000008.tif232159TIFF2025122018000009.tif71161

[0034] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0035] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0036] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0037] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0038] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0039] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0040] In yet another general aspect, the disclosure provides a method for producing a compound having the amino acid sequence: [Table 5] and an amino acid sequence selected from: [Table 6] During the ceremony, Xaa3, Xaa6, Xaa 16 , and Xaa 10 are each independently an α,α-disubstituted amino acid; Xaa1 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa9 is selected from Q, N-methyl Q, E, N-MeGln, and peptoid Q; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a length of 9 to 30 amino acids.

[0041] In some embodiments, Xaa1 is selected from L and Cpa.

[0042] In some embodiments, Xaa9 is selected from Q and N-methylQ.

[0043] In some embodiments, Xaa4 is I.

[0044] In some embodiments, Xaa8 is Q.

[0045] In some embodiments, Xaa5 is R.

[0046] In some embodiments, Xaa1Xaa9 comprises LQ, L(N-methylQ), or (Cpa)(N-methylQ).

[0047] In some embodiments, Xaa4Xaa8Xaa5 comprises IQR.

[0048] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 as well as at least one R.

[0049] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus at least one L.

[0050] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10and Xaa3Xaa4Xaa8Xaa5Xaa6, plus at least one amino acid selected from CBA, Cpa, and Cha.

[0051] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus at least one (2-Nal).

[0052] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus at least one (β-Ala).

[0053] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus HRQR or HRER.

[0054] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus LR, (Cpa)R, or (Cha)R.

[0055] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, as well as (CBA)(2-Nal), (Cpa)(2-Nal), or (Cha)(2-Nal).

[0056] In some embodiments, the polypeptide is selected from: RXaa 16 L(N-methylQ)Xaa 10LRXaa3IQRXaa6(CBA)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 88), RXaa 16 (Cpa)(N-methylQ)Xaa 10 (Cpa)RXaa3IQRXaa6(Cpa)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 89), HRQRXaa 16 LQXaa 10 LRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 90), HRQRXaa 16 LQXaa 10 (Cpa)RXaa3IQRXaa6(Cpa)(2-Nal) (SEQ ID NO: 91), HRQRXaa 16 LQXaa 10 (Cha)RXaa3IQRXaa6(Cha)(2-Nal) (SEQ ID NO: 92), and LEHRERXaa 16 LQXaa 10 LRXaa3IQRXaa6L (sequence number 93).

[0057] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0058] In some embodiments, Xaa3, Xaa6, Xaa 10 , and Xaa 16 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0059] In some embodiments, Xaa3, Xaa6, Xaa 10 , and Xaa 16 are each (S)-2-(4'-pentenyl)alanine.

[0060] In some embodiments, one α-substituent in the α,α-disubstituted amino Xaa3 acid is methyl and the other α-substituent is a first hydrocarbon linker; one α-substituent in the α,α-disubstituted amino Xaa6 acid is methyl and the other α-substituent is a first hydrocarbon linker; α,α-disubstituted amino Xaa 10 One α-substituent in the acid is methyl and the other α-substituent is a second hydrocarbon linker, and an α,α-disubstituted amino Xaa 16 One alpha substituent therein is methyl and the other alpha substituent is a second hydrocarbon linker.

[0061] In some embodiments, the first hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, The second hydrocarbon linker has the formula: [ka] In the formula, one [ka] is Xaa of the second hydrocarbon linker 10 indicates the point of attachment to the α carbon atom of [ka] is Xaa of the second hydrocarbon linker 16 indicates the point of attachment to the α carbon atom of

[0062] In some embodiments, the first hydrocarbon crosslinker has the formula: [ka] The second hydrocarbon crosslinker has the formula: [ka]

[0063] In yet another general aspect, the disclosure provides a method for producing a compound having the amino acid sequence: [Table 7] and an amino acid sequence selected from: [Table 8] During the ceremony, Xaa3, Xaa6, Xaa9, and Xaa 14 are each independently an α,α-disubstituted amino acid; Xaa 15 is selected from R, homoR, and N-methylR; Xaa 16 is selected from S and T, Xaa1 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a length of 9 to 30 amino acids.

[0064] In some embodiments, Xaa 15 is R.

[0065] In some embodiments, Xaa 16 is S.

[0066] In some embodiments, Xaa1 is L.

[0067] In some embodiments, Xaa 15 Xaa 16 Xaa1 contains the RSL.

[0068] In some embodiments, Xaa4 is I.

[0069] In some embodiments, Xaa8 is Q.

[0070] In some embodiments, Xaa5 is R.

[0071] In some embodiments, Xaa4Xaa8Xaa5 comprises IQR.

[0072] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one R.

[0073] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (2-Nal).

[0074] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains HR.

[0075] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains a TLR.

[0076] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains (CBA)(2-Nal) or (4-ClPh)(2-Nal).

[0077] In some embodiments, the polypeptide is selected from: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 94), HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 95), HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-ClPh)(2-Nal) (SEQ ID NO: 96), and HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-Cl-Ph)(2-Nal) (SEQ ID NO: 198), wherein the N-terminus of SEQ ID NO: 95 and SEQ ID NO: 96 is modified with palmitoyl-PEG4.

[0078] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0079] In some embodiments, Xaa3, Xaa6, Xaa9, and Xaa 14are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0080] In some embodiments, Xaa3, Xaa6, Xaa9, and Xaa 14 are each (S)-2-(4'-pentenyl)alanine.

[0081] In some embodiments, one α-substituent in the α,α-disubstituted amino Xaa3 acid is methyl and the other α-substituent is a first hydrocarbon linker; one α-substituent in the α,α-disubstituted amino acid Xaa6 is methyl and the other α-substituent is a first hydrocarbon linker; In the α,α-disubstituted amino Xaa acid, one α-substituent is methyl and the other α-substituent is a second hydrocarbon linker, and the α,α-disubstituted amino acid Xaa 14 One alpha substituent therein is methyl and the other alpha substituent is a second hydrocarbon linker.

[0082] In some embodiments, the first hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, The second hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the second hydrocarbon linker to the α carbon atom of Xaa9, and the other [ka] is Xaa of the second hydrocarbon linker 14 indicates the point of attachment to the α carbon atom of

[0083] In some embodiments, the first hydrocarbon crosslinker has the formula: [ka] The second hydrocarbon crosslinker has the formula: [ka]

[0084] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: [Table 9] Xaa 10 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa2 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit; Xaa3 is selected from D and Nle; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; The polypeptide has a length of 8 to 30 amino acids.

[0085] In some embodiments, Xaa2 is L.

[0086] In some embodiments, Xaa 11 is R.

[0087] In some embodiments, Xaa3 is D.

[0088] In some embodiments, Xaa4 is I.

[0089] In some embodiments, Xaa8 is Q.

[0090] In some embodiments, Xaa5 is R.

[0091] In some embodiments, Xaa2Xaa 11 Xaa3Xaa4Xaa8Xaa5 contains LRDIQR.

[0092] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one L.

[0093] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (2-Nal).

[0094] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (β-Ala).

[0095] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains LQ.

[0096] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains L(2-Nal).

[0097] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (β-Ala).

[0098] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 Xaa3Xaa4Xaa8Xaa5Xaa6 plus HRERS or HRQRS.

[0099] In some embodiments, the polypeptide is: LQXaa 10 LRDIQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 97)

[0100] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0101] In some embodiments, Xaa6 and Xaa 10 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0102] In some embodiments, Xaa6 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine.

[0103] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0104] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, and the other [ka] is Xaa of the hydrocarbon linker 10 indicates the point of attachment to the α carbon atom of

[0105] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0106] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: SXaa1QTXaa2RXaa3Xaa4QXaa5Xaa6Xaa7(2-Nal) (SEQ ID NO: 1), During the ceremony, Xaa1 and Xaa2 are each independently selected from L, A, Cha, Cpa, CBA, (DL), MeL, NMeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; The polypeptide has a length of 13 to 30 amino acids.

[0107] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0108] In some embodiments, Xaa1 and Xaa2 are each L.

[0109] In some embodiments, Xaa1 and Xaa2 are each A.

[0110] In some embodiments, Xaa1 is L and Xaa2 is A.

[0111] In some embodiments, Xaa1 and Xaa2 are each Cha.

[0112] In some embodiments, Xaa1 and Xaa2 are each Cpa.

[0113] In some embodiments, Xaa4 is I and Xaa5 is R.

[0114] In some embodiments, Xaa4 is I and Xaa5 is A.

[0115] In some embodiments, Xaa4 is A and Xaa5 is R.

[0116] In some embodiments, Xaa7 is selected from Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0117] In some embodiments, Xaa7 is L.

[0118] In some embodiments, Xaa7 is A.

[0119] In some embodiments, Xaa7 is CBA.

[0120] In some embodiments, the amino acid sequence SEQ ID NO:1 is selected from the following: SLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 2), and SLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 3).

[0121] In some embodiments, the polypeptide comprises the amino acid sequence RER.

[0122] In some embodiments, the polypeptide comprises the amino acid sequence QER.

[0123] In some embodiments, the polypeptide has a length of 13 to 20 amino acids.

[0124] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:1 plus at least one amino acid selected from Q, L, E, H, and R.

[0125] In some embodiments, the polypeptide is selected from any one of the following polypeptides: RSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 4), RERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 5), HRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 6), EHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 7), QLEHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 8), EHRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 9), QERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 10), HQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 11), EHQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 12), RERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 13), and HRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 14).

[0126] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0127] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0128] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0129] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0130] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0131] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0132] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: Xaa1QTXaa2RXaa3Xaa4QXaa5Xaa6Xaa7(2-Nal) (SEQ ID NO: 15), During the ceremony, Xaa1 and Xaa2 are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; The polypeptide comprises at least one A, The polypeptide has a length of 12 to 30 amino acids.

[0133] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0134] In some embodiments, at least one of Xaa1, Xaa2, Xaa4, Xaa5, and Xaa7 is A.

[0135] In some embodiments, Xaa1 and Xaa2 are each A.

[0136] In some embodiments, Xaa1 is L and Xaa2 is A.

[0137] In some embodiments, Xaa1 and Xaa2 are each L.

[0138] In some embodiments, Xaa2 and Xaa4 are each A.

[0139] In some embodiments, Xaa2 is L and Xaa4 is A.

[0140] In some embodiments, Xaa2 is L and Xaa4 is I.

[0141] In some embodiments, Xaa2 is A and Xaa4 is I.

[0142] In some embodiments, Xaa4 is I and Xaa7 is A.

[0143] In some embodiments, Xaa5 is R and Xaa7 is L.

[0144] In some embodiments, Xaa5 is R and Xaa7 is A.

[0145] In some embodiments, Xaa5 is A and Xaa7 is L.

[0146] In some embodiments, Xaa5 is R and Xaa7 is CBA.

[0147] In some embodiments, Xaa5 is A and Xaa7 is CBA.

[0148] In some embodiments, the amino acid sequence SEQ ID NO:15 is selected from the following: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 16), LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 17), LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 18), LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 19), and LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 20).

[0149] In some embodiments, the polypeptide comprises the amino acid sequence ERS.

[0150] In some embodiments, the polypeptide comprises the amino acid sequence (β-Ala)(β-Ala).

[0151] In some embodiments, the polypeptide comprises the amino acid sequence AA.

[0152] In some embodiments, the polypeptide has a length of 12 to 20 amino acids.

[0153] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:15 plus at least one amino acid selected from Q, L, E, H, R, and S.

[0154] In some embodiments, the polypeptide is selected from any one of the following polypeptides: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 21), LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 22), LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 23), LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 24), LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 25), LQTLRXaa3IQAXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 26), LQTLRXaa3IQAXaa6L(2-Nal)AA (SEQ ID NO: 27), HRERSLQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 28), HRERSLQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 29), LQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 75), LQTLRXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 76), and LQTLRXaa3IQRXaa6A(2-Nal) (sequence number 77).

[0155] In some embodiments, the polypeptide is selected from any one of the following polypeptides: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 21), LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 22), LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 23), LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 24), LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 25), LQTLRXaa3IQAXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 26), LQTLRXaa3IQAXaa6L(2-Nal)AA (SEQ ID NO: 27), HRERSLQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 28), and HRERSLQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 29).

[0156] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0157] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0158] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0159] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0160] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0161] In some embodiments, the hydrocarbon linker has the formula: [ka]

[0162] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: Xaa1QTXaa2RXaa3Xaa4QXaa5Xaa6Xaa7(2-Nal) (SEQ ID NO: 30), During the ceremony, Xaa1 and Xaa2 are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; The polypeptide has a length of 13 to 22 amino acids.

[0163] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0164] In some embodiments, Xaa1 and Xaa2 are each L.

[0165] In some embodiments, Xaa4 is I and Xaa5 is R.

[0166] In some embodiments, Xaa7 is L.

[0167] In some embodiments, the amino acid sequence SEQ ID NO:30 is LQTLRXaa3IQRXaa6L(2-Nal) (sequence number 31).

[0168] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:30, plus at least one amino acid selected from P, D, and β-Ala.

[0169] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:30 plus at least one amino acid selected from Q, L, E, H, R, and S.

[0170] In some embodiments, the polypeptide is selected from any one of the following polypeptides: LQTLRXaa3IQRXaa6L(2-Nal)PD (SEQ ID NO: 32), LQTLRXaa3IQRXaa6L(2-Nal)P (SEQ ID NO: 33), LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34), and LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34a), wherein the C-terminus of SEQ ID NO: 34a is modified with GRKKRRQRRRPQK(PEG4-palmitoyl)NH2.

[0171] In some embodiments, the polypeptide is selected from any one of the following polypeptides: LQTLRXaa3IQRXaa6L(2-Nal)PD (SEQ ID NO: 32), LQTLRXaa3IQRXaa6L(2-Nal)P (SEQ ID NO: 33), LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34).

[0172] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0173] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0174] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0175] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0176] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0177] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0178] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: Xaa1TXaa2RXaa3 (SEQ ID NO: 35), During the ceremony, Xaa1 and Xaa3 are each independently an α,α-disubstituted amino acid; Xaa2 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; the polypeptide contains at least one 2-Nal; The polypeptide has a length of 6 to 30 amino acids.

[0179] In some embodiments, Xaa2 is L or A.

[0180] In some embodiments, Xaa2 is L.

[0181] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:35 plus at least one amino acid selected from Q, L, E, H, I, S, M, and R.

[0182] In some embodiments, the polypeptide comprises at least two R's in addition to the amino acid sequence SEQ ID NO:35.

[0183] In some embodiments, the polypeptide comprises three Rs in addition to the amino acid sequence SEQ ID NO:35.

[0184] In some embodiments, the polypeptide comprises at least two E's in addition to the amino acid sequence SEQ ID NO:35.

[0185] In some embodiments, the polypeptide comprises at least two Ls in addition to the amino acid sequence SEQ ID NO:35.

[0186] In some embodiments, the polypeptide comprises three Ls in addition to the amino acid sequence SEQ ID NO:35.

[0187] In some embodiments, the polypeptide comprises at least two Q's in addition to the amino acid sequence SEQ ID NO:35.

[0188] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:35 plus at least one amino acid selected from H, S, I, and M.

[0189] In some embodiments, the polypeptide comprises the amino acid sequence IQR.

[0190] In some embodiments, the polypeptide comprises the amino acid sequence ML(2-Nal).

[0191] In some embodiments, the polypeptide comprises the amino acid sequence (2-Abu)L(2-Nal)(β-Ala)(β-Ala).

[0192] In some embodiments, the polypeptide comprises the amino acid sequence RERSL.

[0193] In some embodiments, the polypeptide comprises the amino acid sequence QLEH.

[0194] In some embodiments, the polypeptide is selected from: QLEHRERSLXaa1TLRXaa3IQRML(2-Nal) (SEQ ID NO: 36), and QLEHRERSLXaa1TLRXaa3IQR(2-Abu)L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 78).

[0195] In some embodiments, the polypeptide is QLEHRERSLXaa1TLRXaa3IQRML(2-Nal) (sequence number 36).

[0196] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0197] In some embodiments, Xaa1 and Xaa3 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0198] In some embodiments, Xaa1 and Xaa3 are each (S)-2-(4'-pentenyl)alanine.

[0199] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0200] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3.

[0201] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0202] In yet another general aspect, the disclosure provides a method for detecting a nucleotide sequence comprising administering to a subject a subject an amino acid sequence selected from the following: Xaa1SLQXaa2 (SEQ ID NO: 37a) and Xaa1S(Cha)(N-methylQ)Xaa2 (SEQ ID NO: 37b), and an amino acid sequence selected from: Xaa3IQRXaa4 (SEQ ID NO: 38a) and Xaa3IQQXaa4 (SEQ ID NO: 38b), or a pharmaceutically acceptable salt thereof, During the ceremony, Xaa1, Xaa2, Xaa3, and Xaa4 are each independently an α,α-disubstituted amino acid; The polypeptide has a length of 10 to 30 amino acids.

[0203] In some embodiments, the polypeptide has the amino acid sequence: Xaa1SLQXaa2 (SEQ ID NO: 37), and Amino acid sequence: Contains Xaa3IQRXaa4 (SEQ ID NO: 38).

[0204] In some embodiments, the polypeptide comprises at least one 2-Nal.

[0205] In some embodiments, the polypeptide comprises at least one CBA.

[0206] In some embodiments, the polypeptide comprises, in addition to the amino acid sequence SEQ ID NO:37a or SEQ ID NO:37b, and the amino acid sequence SEQ ID NO:38a or SEQ ID NO:38b, at least one amino acid selected from L, E, R, H, Q, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, homoHis, NHis, homoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylic Gly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

[0207] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:37a or SEQ ID NO:37b, and the amino acid sequence SEQ ID NO:38a or SEQ ID NO:38b, as well as at least one amino acid selected from L, E, R, H, Q, N-methyl E, CBA, N-methyl Q, Cha, and N-methyl R.

[0208] In some embodiments, the polypeptide comprises at least one amino acid selected from L, E, and R in addition to the amino acid sequence SEQ ID NO:37 and the amino acid sequence SEQ ID NO:38.

[0209] In some embodiments, the polypeptide comprises the amino acid sequence RE.

[0210] In some embodiments, the polypeptide comprises the amino acid sequence LR.

[0211] In some embodiments, the polypeptide comprises the amino acid sequence L(2-Nal).

[0212] In some embodiments, the polypeptide comprises the amino acid sequence (CBA)(2-Nal).

[0213] In some embodiments, the polypeptide comprises an amino acid sequence selected from HRE, HR(N-methyl E), HR(N-methyl Q), HRQ, LR, L(N-methyl R), (Cha)R, L(2-Nal), and (CBA)(2-Nal).

[0214] In some embodiments, the polypeptide comprises at least one β-Ala.

[0215] In some embodiments, the polypeptide has a length of 10 to 20 amino acids.

[0216] In some embodiments, the polypeptide is selected from any one of the following polypeptides: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal) (SEQ ID NO: 39), REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 40), EXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 79), HREXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 82), HREXaa1SLQXaa2LRXaa3IQQXaa4(CBA)(2-Nal) (SEQ ID NO: 83), HR(N-methyl E)Xaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 84), HREXaa1SLQXaa2L(N-methylR)Xaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 85), HR(N-methylQ)Xaa1S(Cha)(N-methylQ)Xaa2(Cha)RXaa3IQRXaa4(Cha)(2-Nal) (SEQ ID NO: 86), and HRQXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 87).

[0217] In some embodiments, the polypeptide is selected from any one of the following polypeptides: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal) (SEQ ID NO: 39), REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 40).

[0218] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0219] In some embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0220] In some embodiments, Xaa1 and Xaa2 are each (S)-2-(4'-pentenyl)alanine.

[0221] In some embodiments, Xaa3 and Xaa4 are each (S)-2-(4'-pentenyl)alanine.

[0222] In some embodiments, one α-substituent in the α,α-disubstituted amino acid Xaa1 is methyl and the other α-substituent is a first hydrocarbon linker, and one α-substituent in the α,α-disubstituted amino acid Xaa2 is methyl and the other α-substituent is a first hydrocarbon linker.

[0223] In some embodiments, the first hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the first hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the first hydrocarbon linker to the α carbon atom of Xaa2.

[0224] In some embodiments, the first hydrocarbon crosslinker has the formula: [ka]

[0225] In some embodiments, one α-substituent in the α,α-disubstituted amino acid Xaa3 is methyl and the other α-substituent is a second hydrocarbon linker, and one α-substituent in the α,α-disubstituted amino acid Xaa4 is methyl and the other α-substituent is a second hydrocarbon linker.

[0226] In some embodiments, the second hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the second hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the second hydrocarbon linker to the α carbon atom of Xaa4.

[0227] In some embodiments, the second hydrocarbon crosslinker has the formula: [ka]

[0228] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence SEQ ID NO:37, or a pharmaceutically acceptable salt thereof. [Table 10] During the ceremony, Xaa 15 and Xaa 10 are each independently an α,α-disubstituted amino acid; the polypeptide comprises at least two Nle; The polypeptide has a length of 7 to 30 amino acids.

[0229] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:37, as well as at least one amino acid selected from L, E, R, H, S, Q, I, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, HomoHis, NHis, HomoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylic Gly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

[0230] In some embodiments, the polypeptide comprises, in addition to the amino acid sequence SEQ ID NO: 37, at least one amino acid selected from the following: H, R, E, S, L, Q, I, CBA, and (2-Nal).

[0231] In some embodiments, the polypeptide comprises an IQR.

[0232] In some embodiments, the polypeptide comprises (Nle)IQR(Nle).

[0233] In some embodiments, the polypeptide comprises an HRE and a LR.

[0234] In some embodiments, the polypeptide comprises L(2-Nal).

[0235] In some embodiments, the polypeptide comprises (CBA)(2-Nal).

[0236] In some embodiments, the polypeptide is selected from: HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)L(2-Nal) (SEQ ID NO: 80), and HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)(CBA)(2-Nal) (SEQ ID NO: 81).

[0237] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0238] In some embodiments, Xaa 15 and Xaa 10 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0239] In some embodiments, Xaa 15 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine.

[0240] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0241] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] is Xaa of the hydrocarbon linker 15 indicates the point of attachment to the α carbon atom of [ka] is Xaa of the hydrocarbon linker 10 indicates the point of attachment to the α carbon atom of

[0242] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0243] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: [Table 11] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit, Xaa 11and at least one of Xaa5 contains E or K; The polypeptide has a length of 8 to 30 amino acids.

[0244] In some embodiments, Xaa 11 is E and Xaa5 is R.

[0245] In some embodiments, Xaa 11 is R and Xaa5 is E.

[0246] In some embodiments, Xaa 11 is K and Xaa5 is R.

[0247] In some embodiments, Xaa 11 is R and Xaa5 is K.

[0248] In some embodiments, the polypeptide comprises L(2-Nal).

[0249] In some embodiments, the polypeptide comprises IQR, IQE, or IQK.

[0250] In some embodiments, the polypeptide comprises LQTLE, LQTLR, or LQRLK.

[0251] In some embodiments, selected from: LQTLEXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 98), LQTLRXaa3IQEXaa6L(2-Nal) (SEQ ID NO: 99), LQTLKXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 100), and LQTLRXaa3IQKXaa6L(2-Nal) (sequence number 101).

[0252] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0253] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0254] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0255] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0256] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0257] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0258] In yet another general aspect, the disclosure provides a polypeptide comprising the amino acid sequence: [Table 12] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh, wherein The polypeptide comprises at least one of (DI) or (DL), The polypeptide has a length of 7 to 30 amino acids.

[0259] In some embodiments, Xaa4 is (DI).

[0260] In some embodiments, Xaa7 is (DL).

[0261] In some embodiments, the polypeptide comprises a (DL)QTIR.

[0262] In some embodiments, the polypeptide comprises LQT(DL)R.

[0263] In some embodiments, the polypeptide is selected from: (DL)QTIRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 102), LQT(DL)RXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 103), LQTLRXaa3(DI)QRXaa6L(2-Nal) (SEQ ID NO: 104), and LQTLRXaa3IQRXaa6(DL)(2-Nal) (SEQ ID NO: 105).

[0264] In some embodiments, the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

[0265] In some embodiments, Xaa3 and Xaa6 are each independently selected from the following: (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

[0266] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0267] In some embodiments, one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

[0268] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0269] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0270] In yet another general aspect, the disclosure provides a polypeptide having a length of 6 to 30 amino acids, or a pharmaceutically acceptable salt thereof, wherein the polypeptide has at least 60% homology to a corresponding fragment of the wild-type HD2 domain of human B-cell CLL / lymphoma 9 (BCL9), and wherein the polypeptide comprises at least one α-monosubstituted unnatural amino acid.

[0271] In some embodiments, the fragment of the wild-type HD2 domain of human B-cell CLL / lymphoma 9 (BCL9) is any fragment between positions 355 and 377 within BCL9.

[0272] In some embodiments, the α-monosubstituted unnatural amino acid is selected from Nle, β-Ala, 2-Nal, β-L, and CBA.

[0273] In some embodiments, the polypeptide comprises at least one 2-Nal.

[0274] In some embodiments, the polypeptide comprises at least one CBA.

[0275] In some embodiments, the polypeptide comprises at least one Nle.

[0276] In some embodiments, the polypeptide comprises two Nle.

[0277] In some embodiments, the polypeptide comprises at least one β-L.

[0278] In some embodiments, the polypeptide comprises at least one β-Ala.

[0279] In some embodiments, the polypeptide comprises two β-Ala.

[0280] In some embodiments, the polypeptide comprises at least one amino acid sequence selected from the following: DIQRML(2-Nal) (SEQ ID NO: 41), (Nle)IQR(Nle)L(2-Nal) (SEQ ID NO: 42), (Nle)IQR(Nle)(CBA)(2-Nal) (SEQ ID NO: 43), (Nle)IQA(Nle)L(2-Nal) (SEQ ID NO: 44), (Nle)IQA(Nle)(CBA)(2-Nal) (SEQ ID NO: 45), (Nle)TLR(Nle) (SEQ ID NO: 46), QTLR (Nle) (SEQ ID NO: 47), and QT(β-L)R(Nle) (SEQ ID NO: 48).

[0281] In some embodiments, the polypeptide comprises the amino acid sequence RSL.

[0282] In some embodiments, the polypeptide comprises an amino acid sequence selected from an HRE and an HQE.

[0283] In some embodiments, the polypeptide comprises the amino acid sequence QLE.

[0284] In some embodiments, the polypeptide is selected from any one of the following polypeptides: [Table 13]

[0285] In yet another general aspect, the disclosure provides a pharmaceutically acceptable composition comprising any one of the polypeptides disclosed herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0286] In yet another general aspect, the disclosure provides a method comprising: a method for inhibiting the binding of BCL9 to β-catenin in a subject; and / or a method of inhibiting canonical Wnt / β-catenin signaling in a subject; and / or a method for decreasing the survival of regulatory T cells in a subject; and / or a method for reducing the expression of VEGF in a tumor in a subject; and / or a method for increasing the infiltration of CD4+ T cells and CD8+ T cells into a tumor in a subject; and / or a method for increasing T helper 17 (Th17) cells to tumors in a subject; and / or a method for reducing intratumoral dendritic cells in a subject; and / or A half-life (T) of at least 2 hours when administered to a subject 1 / 2 ) and / or a method for inducing a tumor microenvironment favorable for an immune response in a subject; and / or a method for inhibiting tumor growth in a subject, and / or a method for inhibiting cancer stem cell proliferation in a subject; and / or a method for inhibiting tumor metastasis in a subject, and / or ● A method of treating cancer in a subject; Methods are provided that include administering to a subject in need thereof a therapeutically effective amount of any one of the polypeptides disclosed herein, or a pharmaceutical composition comprising any one of the polypeptides.

[0287] In some embodiments, the cancer is familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, kidney cancer, brain / CNS cancer, throat cancer, multiple myeloma, cutaneous melanoma, acute lymphocytic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangioma, gastric cancer, ovarian cancer, hepatocellular carcinoma, or lymphangiogenesis.

[0288] In some embodiments, the cancer is colorectal cancer.

[0289] In some embodiments, the cancer is gastric cancer.

[0290] In some embodiments, the cancer is ovarian cancer.

[0291] In some embodiments, the cancer is hepatocellular carcinoma.

[0292] In some embodiments, the cancer is breast cancer.

[0293] In some embodiments, the cancer is prostate cancer.

[0294] In some embodiments, the cancer is cutaneous melanoma.

[0295] In some embodiments, the cancer is lung cancer.

[0296] In some embodiments, the method further comprises administering at least one additional therapeutic agent to the subject.

[0297] In some embodiments, the at least one additional agent is selected from the group consisting of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a chemotherapeutic agent, and a VEGFR inhibitor.

[0298] In some embodiments, the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody.

[0299] In some embodiments, the checkpoint inhibitor targets a stimulatory checkpoint molecule selected from the group consisting of CD27, CD40, OX40, GITR, and CD137.

[0300] In some embodiments, the checkpoint inhibitor targets an inhibitory checkpoint molecule selected from the group consisting of A2AR, B7-H3, B7-H4, attenuator of B and T lymphocytes (BTLA), indoleamine 2,3-dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG3), T-cell immunoglobulin and mucin domain 3 (TIM-3), VISTA (C10orf54), and V-domain Ig suppressor of T-cell activation.

[0301] In some embodiments, the EGFR inhibitor is erlotinib, gefitinib, lapatinib, panitumumab, vandetanib, or cetuximab.

[0302] In some embodiments, the VEGF inhibitor or VEGFR inhibitor is pazopanib, bevacizumab, sorafenib, sunitinib, axitinib, ponatinib, regorafenib, vandetanib, cabozantinib, ramucirumab, lenvatinib, or ziv-aflibercept.

[0303] In some embodiments, the chemotherapeutic agent is cyclophosphamide, methotrexate, 5-fluorouracil (5-FU), doxorubicin, mustine, vincristine, procarbazine, prednisolone, dacarbazine, bleomycin, etoposide, cisplatin, epirubicin, capecitabine, folinic acid, actinomycin, all-trans retinoic acid, azacitidine, azathioprine, bortezomib, carboplatin, The drugs are acetaminophen, chlorambucil, cytarabine, daunorubicin, docetaxel, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, paclitaxel, pemetrexed, teniposide, thioguanine, topotecan, valrubicin, vinblastine, vindesine, vinorelbine, or oxaliplatin.

[0304] In some embodiments, the method further comprises exposing the subject to radiation therapy and / or chemotherapy.

[0305] In some embodiments, the methods further comprise measuring at least one biomarker to monitor treatment / inhibition effectiveness and / or to select subjects for treatment.

[0306] In some embodiments, the biomarker is one or more of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin.

[0307] In some embodiments, a decrease in gene expression and / or protein levels of CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin indicates therapeutic / inhibitory efficacy, and / or the subject is selected for treatment if the gene expression and / or protein levels of CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin are elevated.

[0308] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this application pertains. Methods and materials are described herein for use in this application; other suitable methods and materials known in the art can also be used. Materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0309] Other features and advantages of the present application will be apparent from the following detailed description and drawings, and from the claims. [Brief explanation of the drawings]

[0310] [Figure 1] 1 shows a biacore assay to assess β-catenin binding of stapled peptides or SEQ ID NO: 147. [Figure 2] 1 shows the pharmacokinetic profile of stapled peptide SEQ ID NO:79. DETAILED DESCRIPTION OF THE INVENTION

[0311] BCL-9, β-catenin, and Wnt signaling Aberrant activation of Wnt signaling has been implicated in various cancers, as tumors can become dependent on Wnt signaling for growth and survival (see, e.g., Grossmann et al. PNAS. 109(44):17942-17947 (2012)). Up to 90% of all sporadic colorectal cancer cases are associated with constitutive activation of Wnt signaling.

[0312] β-catenin is a protein that may be involved in protein-protein interactions that stimulate Wnt signaling, leading to changes in transcriptional activation that can enable tumor growth and development. β-catenin is normally phosphorylated and targeted for degradation by the Axin complex. When the Wnt signaling pathway is stimulated, unphosphorylated β-catenin accumulates, binds to lymphoid enhancer factor / T cell factor (LEF / TCF), translocates to the nucleus, and stimulates the transcription of Wnt target genes (see, e.g., Thakur and Mishra, J Cell Mol Med 17(4):449-456(2013)). Wnt target genes include c-myc and CD44, which are upregulated in tumor models. BCL9 is a protein required for efficient β-catenin-mediated transcription in mammalian cells (see, e.g., de la Roche et al., BMC Cancer 8:199(2008)).

[0313] "Canonical" Wnt / β-catenin signaling is a pathway activated by Wnt ligands binding to the Frizzled family of cell surface receptors, which then regulate the expression and subcellular localization of β-catenin. In the absence of Wnt ligands, β-catenin is phosphorylated and ubiquitinated within a destruction complex composed of apoptosis coli (APC), glycogen synthase kinase 3 (GSK-3), casein kinase 1 (CK1), and Axin, and targeted for degradation in a proteasome-dependent manner. In the presence of Wnt ligands, β-catenin ubiquitination in this complex is inhibited, saturating phosphorylated β-catenin, which then stabilizes and translocates to the nucleus. Within the nucleus, phosphorylated β-catenin recruits nuclear T-cell factor (TCF) transcription factors, such as lymphoid enhancer factor / 3 (LEF / TCF), to induce the expression of genes, including c-Myc28 and cyclin D, that promote cell proliferation, migration, and survival.

[0314] Several molecules, including BCL9 and its homologue B-cell lymphoma 9-like (B9L), have been shown to be coactivators for Wnt / β-catenin transcription. The formation of a complex consisting of TCF, β-catenin, and BCL9 (or B9L) enhances β-catenin-dependent Wnt transcriptional activity. In normal cells, this transcriptional pathway is blocked when Wnt ligands dissociate from their receptors. However, various loss-of-function mutations in APC and Axin, as well as activating mutations in β-catenin itself, allow β-catenin to escape the destruction complex and accumulate in the nucleus. Such inappropriate persistence of β-catenin promotes the oncogenesis of a wide range of common human epithelial cancers, including hepatocellular, breast, colorectal, and hematologic malignancies, such as multiple myeloma. In addition, active β-catenin signaling is essential for the development of T cells, particularly CD8 +This leads to the elimination of T cells, which leads to treatment resistance and shortened patient survival. Therefore, blocking Wnt signaling by targeting β-catenin (β-cat) may represent a powerful CRC treatment that may prevent both tumor initiation and metastasis. See, for example, Spranger et al., Nature 523:231-235 (2015).

[0315] As with other transcription factors, the development of selective, nontoxic β-catenin inhibitors and their translation into clinical practice has proven a considerable challenge, as β-catenin interacts with most of its protein partners via the same binding surface. Consequently, Wnt pathway inhibitors targeting this common binding surface have exhibited significant adverse effects in animal studies and clinical trials. Only a few drugs targeting β-catenin are currently in clinical trials, including PRI-724 (Eisai Pharmaceuticals; Phase II), LGK974 (Novartis; Phase I), and OMP-54F28 and OMP-18R5 (OncoMed / Bayer; Phase I). Additionally, disruption of the LEF / TCF interaction by small molecule and peptide inhibitors of β-catenin can cause severe side effects in treated mice, including severe bone marrow hypoplasia, anemia, and generalized wasting, likely resulting from disruption of homeostatic Wnt signaling in normal hematopoietic and intestinal stem cells. Such therapeutic limitations may stem from disruption of the interactions between β-catenin and TCF and β-catenin and E-cadherin, which may affect epithelial tissue integrity. Furthermore, biologic agents targeting Frizzled receptors (OMP-54F28 and OMP-18R5) have shown significant bone marrow toxicity during clinical trials. Although Wnt ligands are essential for Wnt / β-catenin activation, APC and β-catenin mutations in cancer cells can induce downstream transcription without Wnt ligand activation. Therefore, blocking Wnt secretion fails to inhibit endogenous oncogenic Wnt activity due to downstream gene transcription induced by APC and β-catenin mutations. LGK974 targets only a small patient population, as identified by certain biomarkers. The small molecule inhibitor PRI-724 is currently in phase II trials using daily infusions, but intravenous (IV) dosing more than once a week presents undesirable and unsupported characteristics for clinical development.

[0316] Traditionally, the Wnt signaling pathway includes three distinct types of signaling: the canonical Wnt signaling pathway, in which Wnt regulates various transcriptional target genes in a β-catenin-dependent manner; the non-canonical Wnt signaling pathway, in which Wnt is primarily involved in planar cell polarity and can function independently of β-catenin; and the non-canonical Wnt / calcium pathway, in which Wnt regulates intracellular calcium levels. In this application, "canonical Wnt signaling" is interchangeably referred to as "canonical Wnt / β-catenin signaling" or "Wnt signaling." As described herein, canonical Wnt / β-catenin signaling may refer to pathway components that regulate the amount of β-catenin in a patient or sample, for example, by regulating β-catenin stability. In some embodiments, canonical Wnt / β-catenin signaling includes pathway components that transcriptionally regulate one or more genes, such as c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2, and LEF1. In some embodiments, canonical Wnt / β-catenin signaling includes pathway components regulated by the interaction of β-catenin with BCL9. In some embodiments, canonical Wnt / β-catenin signaling includes one or more genes that are transcriptionally regulated by the interaction of β-catenin with BCL9. The one or more genes regulated by the interaction of β-catenin with BCL9 may include c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2, and LEF1. In some embodiments, canonical Wnt / β-catenin signaling includes one or more proteins whose transcriptional expression is regulated by the interaction of β-catenin with BCL9. These components may include, for example, c-myc, cyclin D1, CD44, LGR5, VEGFA, AXIN2, and LEF1.

[0317] Polypeptides derived from the BCL9 HD2 domain The HD2 domain of the BCL9 protein mediates BCL9 binding to β-catenin, and to date, the HD2 domain is the only domain of BCL9 shown to bind to β-catenin intracellularly (see, e.g., de la Roche 2008). The human BCL9 protein has an amino acid sequence of approximately 1426 amino acids (GeneID 607). In some embodiments, the present application provides polypeptides comprising at least a fragment of the wild-type HD2 domain of BCL9. The full-length HD2 domain of BCL9 comprises a sequence of 30 amino acids corresponding to positions 348-377 in the BCL9 protein. The sequence of the full-length HD2 domain of the human BCL9 protein (SEQ ID NO: X) is set forth in Table 1, with numerical references to the corresponding amino acid positions in the BCL9 protein. [Table 14]

[0318] With reference to Table 1, P at position 348 is the N-terminal amino acid, and E at position 377 is the C-terminal amino acid. In some embodiments, the polypeptides described herein comprise the full-length HD2 domain of a human BCL9 protein. In some embodiments, the polypeptides described herein comprise a fragment of the HD2 domain of a human BCL9 protein, or a variant thereof. In some embodiments, the fragment of the HD2 domain of a human BCL9 protein, or a variant thereof, is 6-30 amino acids in length. In some embodiments, the polypeptides are 7-14 amino acids, 9-14 amino acids, 7-12 amino acids, 10-14 amino acids, 6-20 amino acids, 7-20 amino acids, 9-20 amino acids, 10-20 amino acids, 11-20 amino acids, 12-20 amino acids, 12-30 amino acids, 13-20 amino acids, 13-22 amino acids, or 14-19 amino acids in length. For example, the polypeptide has 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids. In some embodiments, the polypeptide comprises (or consists of) any fragment of the wild-type HD2 domain of the BCL9 protein between positions 355 and 377, inclusive, within the BCL9 protein. For example, the polypeptide may comprise the sequence of 14 amino acids between positions 363 and 376, or the sequence of 14 amino acids between positions 361 and 374, or the sequence of 16 amino acids between positions 359 and 374, or the sequence of 17 amino acids between positions 358 and 374, or the sequence of 18 amino acids between positions 357 and 374, or the sequence of 20 amino acids between positions 355 and 374, inclusive, within the BCL9 protein, or a variant thereof. In some embodiments, the polypeptide comprises (or consists of) a fragment of the HD2 domain corresponding to the sequence of amino acids 355 to 376 in the BCL9 protein, or a variant thereof.In some embodiments, the polypeptide comprises (or consists of) a fragment of the HD2 domain corresponding to the amino acid sequence of positions 366-376, 366-374, 363-374, 363-376, 363-375, 361-374, 359-374, 358-374, 357-374, or 358-376 within the BCL9 protein, or a variant thereof.

[0319] In certain embodiments, the variants described herein are polypeptides derived from the HD2 domain of the human BCL9 protein, including fragments of the HD2 domain of the human BCL9 protein, further modified by substituting one or more amino acids with other naturally occurring or non-naturally occurring amino acids. In some embodiments, the variants comprise conservative substitutions of one or more amino acids in the HD2 domain of the BCL9 protein, or a fragment thereof. In some embodiments, at least one, at least two, at least three, at least four, at least five, at least six, or at least seven amino acids in the HD2 domain or a fragment thereof are substituted with different amino acids. Conservative amino acid substitutions, i.e., replacing an amino acid with a different amino acid with similar properties (e.g., hydrophilicity and degree and distribution of charged regions), usually do not significantly alter the biological activity of the polypeptide because the changes involved are minor. Such minor changes can be identified by considering the hydropathic index of the amino acid, based on consideration of the amino acid's hydrophobicity and charge. Amino acids with similar hydropathic indexes and hydrophilicity values can be substituted to retain protein function. Both the hydrophobicity index and hydrophilicity value of an amino acid are influenced by the particular side chain of that amino acid. In keeping with this observation, amino acid substitutions that are compatible with biological function depend on the relative similarity of amino acids, as revealed by their hydrophobicity, hydrophilicity, charge, size, and other properties, and particularly on the side chains of those amino acids.

[0320] In some embodiments, the variant polypeptides described herein are at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% identical to the corresponding fragment of the wild-type HD2 domain of the BCL9 protein. In some embodiments, the polypeptides are variants that are at least 60%, at least 70%, or at least 80% identical to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 363 to 374, inclusive, in the BCL9 protein. In some embodiments, the polypeptides are variants that are at least 60%, at least 70%, or at least 80% identical to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 363 to 376, inclusive, in the BCL9 protein. In some embodiments, the polypeptides are variants that are at least 60%, at least 70%, or at least 80% identical to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 363 to 375, inclusive, in the BCL9 protein. In some embodiments, the polypeptide is a variant having at least 60%, at least 70%, or at least 80% homology to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 361 to 374, inclusive, in the BCL9 protein. In some embodiments, the polypeptide is a variant having at least 60%, at least 70%, or at least 80% homology to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 359 to 374, inclusive, in the BCL9 protein. In some embodiments, the polypeptide is a variant having at least 60%, at least 70%, or at least 80% homology to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 358 to 374, inclusive, in the BCL9 protein. In some embodiments, the polypeptide is a variant having at least 60%, at least 70%, or at least 80% homology to a fragment of the HD2 domain corresponding to the amino acid sequence of positions 357 to 374, inclusive, in the BCL9 protein.In some embodiments, the polypeptide is a variant having at least 60%, at least 70%, or at least 80% homology to a fragment of the HD2 domain corresponding to amino acid sequence from positions 355 to 374, inclusive, in the BCL9 protein.

[0321] In some embodiments, the variant comprises a substitution of L with A (e.g., at a position corresponding to position 373 of the BCL9 protein). In some embodiments, the variant comprises a substitution of L with A (e.g., at a position corresponding to position 363 or 366 of the BCL9 protein). In some embodiments, the variant comprises a substitution of I with A (e.g., at a position corresponding to position 369 of the BCL9 protein). In some embodiments, the variant comprises a substitution of P with A (e.g., at a position corresponding to position 375 of the BCL9 protein). In some embodiments, the variant comprises a substitution of D with A (e.g., at a position corresponding to position 376 of the BCL9 protein). Such subtle changes in amino acid hydrocarbon side chains modify (e.g., reduce) the overall lipophilicity of the polypeptide.

[0322] In some embodiments, the variant comprises a substitution of R with A (e.g., at a position corresponding to position 371 of the BCL9 protein). In some embodiments, the variant comprises a substitution of R with Q (e.g., at a position corresponding to position 359 of the BCL9 protein). Removal of at least one R bearing a charged guanidinium moiety reduces the net positive charge of the polypeptide (e.g., the net positive charge can be reduced by -1, -2, or -3, such that the total charge of the polypeptide including R, H, and / or L is +2 or +3).

[0323] Polypeptides containing α-monosubstituted unnatural amino acids In some embodiments, the variant comprises a substitution of one or more amino acids (e.g., 1, 2, 3, or 4 amino acids) of the HD2 domain of a BCL9 protein, or a corresponding fragment thereof, with a non-naturally occurring amino acid. In some embodiments, the non-naturally occurring amino acid is an α-monosubstituted unnatural amino acid.

[0324] In some embodiments, the α-mono substituted unnatural amino acid is Nle, β-Ala, 2-Nal, β-L, or CBA. In some embodiments, the variant includes at least one Nle. In some embodiments, the variant includes at least one 2-Nal. In some embodiments, the variant includes at least one β-Ala. In some embodiments, the variant includes at least one β-L. In some embodiments, the variant includes at least one CBA. In some embodiments, the variant includes two Nles. In some embodiments, the variant includes two β-Alas. In some embodiments, the variant includes two Nles and one 2-Nal. In some embodiments, the variant includes two Nles, two β-Alas, and one 2-Nal. In some embodiments, the variant includes two Nles, one CBA, and one 2-Nal. In some embodiments, the variant includes two Nles, one β-L, and one 2-Nal. In some embodiments, the variant comprises one CBA and one 2-Nal.

[0325] In some embodiments, the variant comprises a substitution of F with 2-Nal (e.g., at a position corresponding to position 374 of the BCL9 protein). In some embodiments, the variant comprises a substitution of L with CBA (e.g., at a position corresponding to position 373 of the BCL9 protein). In some embodiments, the variant comprises a substitution of L with β-L (e.g., at a position corresponding to position 366 of the BCL9 protein). In some embodiments, the variant comprises a substitution of Q with Nle (e.g., at a position corresponding to position 364 of the BCL9 protein). In some embodiments, the variant comprises a substitution of D with Nle (e.g., at a position corresponding to position 368 of the BCL9 protein). In some embodiments, the variant comprises a substitution of M with Nle (e.g., at a position corresponding to position 372 of the BCL9 protein).

[0326] In some embodiments, the variants include one D substituted with Nle, one M substituted with Nle, one L substituted with CBA, and one F substituted with 2-Nal. In some aspects of these embodiments, the variants also include one R substituted with A. In some embodiments, the variants include one R substituted with Q, one D substituted with Nle, one M substituted with Nle, and one F substituted with 2-Nal. In some embodiments, the variants include one R substituted with A, one D substituted with Nle, one M substituted with Nle, and one F substituted with 2-Nal. In some embodiments, the variants include one F substituted with 2-Nal, one P substituted with β-Ala, and one D substituted with β-Ala. In some aspects of these embodiments, the variants also include one R substituted with A, one D substituted with Nle, and one M substituted with Nle. In some embodiments, the variants include one L substituted with β-L, one D substituted with Nle, one M substituted with Nle, and one F substituted with 2-Nal. In some aspects of these embodiments, the variants also include one R substituted with Q. In some embodiments, the variants include one L substituted with A and one F substituted with 2-Nal. In some aspects of these embodiments, the variants include one I substituted with A. In some embodiments, the variants include one I substituted with A and one F substituted with 2-Nal. In some aspects of these embodiments, the variants include one L substituted with A. In some embodiments, the variants include two Ls, each substituted with A, and one F substituted with 2-Nal. In some embodiments, the variants include one R substituted with A and one F substituted with 2-Nal. In some aspects of these embodiments, the variant also includes one P replaced by A and one D replaced by A. In other aspects of these embodiments, the variant includes one L replaced by CBA.

[0327] In some embodiments, the polypeptides described herein comprise at least one amino acid sequence listed in Table 2. [Table 15]

[0328] In some embodiments, any of the polypeptides disclosed in the present application comprises at least one amino acid sequence listed in Table 3. [Table 16]

[0329] In some embodiments, the polypeptide of the present disclosure is selected from any one of the polypeptides listed in Table 4. [Table 17]

[0330] Non-stapled polypeptides containing α,α-disubstituted amino acids In some embodiments, a polypeptide derived from the HD2 domain of a human BCL9 protein can undergo a reaction to form a hydrocarbon linker between two amino acids in the backbone of the polypeptide. As used herein, a polypeptide capable of undergoing a reaction to form one or more hydrocarbon linkers can be referred to as a "non-stapled polypeptide." In these embodiments, the polypeptide comprises at least two α,α-disubstituted amino acids. In some embodiments, at least one α-substituent in each α,α-disubstituted amino acid in the peptide backbone comprises a double bond. Thus, a polypeptide comprising at least two α,α-disubstituted amino acids can undergo a metathesis reaction to form a hydrocarbon linker between the two α,α-disubstituted amino acids in the peptide backbone.

[0331] In some embodiments, the non-naturally occurring α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid. In certain embodiments, the non-naturally occurring amino acid is a chiral molecule containing a chiral center having either the S or R configuration. In some embodiments, the non-naturally occurring amino acid is selected from the following: [ka] [ka] [ka] [ka] .

[0332] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence shown below: [Table 18]

[0333] In some embodiments, Xaa1-Xaa 16any one of which is independently an α,α-disubstituted amino acid (e.g., an α-methyl, α-alkenyl amino acid described herein), Q, L, E, H, R, E, R, S, L, Q, T, L, R, D, I, Q, R, M, L, F, P, D, E, 2-Nal, Nle, β-Ala, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, N-methyl L, Cpa, Cha, N-MeHis, N-MeCys, homoHis, NHis, homoR , Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylGly, Alg, AC4C, A6C, Aze, (β-t-Bu-Ala), Tle, peptoid Q, DThr, NMeLeu, beta homoTrp, homoCha, Lys(Me)2, Narg, Abg, Nar, Hyp, and Ngln.

[0334] In some embodiments, Xaa1-Xaa 16 any one of which is independently an α,α-disubstituted amino acid (e.g., an α-methyl, α-alkenyl amino acid described herein), Q, L, E, H, R, E, R, S, L, Q, T, L, R, D, I, Q, R, M, L, F, P, D, E, 2-Nal, Nle, β-Ala, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, N-methyl L, Cpa, Cha, N-MeHi s, N-MeCys, homoHis, NHis, homoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methylL, allylGly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

[0335] Xaa1 In some embodiments, Xaa1 is selected from L, MeL, AC4C, A6C, Aze, Cpa, Cha, NMeCha, Dcha, Phe(4-Cl), (β-tBu-Ala), Tle, NMeLeu, beta homoTrp, homoCha, 4-ClPh, 4-FPh, 3,4-diClPh, and NptGly.

[0336] In some embodiments, Xaa1 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly.

[0337] In some embodiments, Xaa1 is L. In some embodiments, Xaa1 is A. In some embodiments, Xaa1 is Cha. In some embodiments, Xaa1 is Cpa. In some embodiments, Xaa1 is (DL). In some embodiments, Xaa1 is CBA. In some embodiments, Xaa1 is MeL. In some embodiments, Xaa1 is NMeCha. In some embodiments, Xaa1 is Dcha. In some embodiments, Xaa1 is NptGly.

[0338] Xaa2 In some embodiments, Xaa2 is selected from L, MeL, AC4C, A6C, Aze, Cpa, Cha, NMeCha, Dcha, Phe(4-Cl), (β-tBu-Ala), Tle, NMeLeu, beta homoTrp, homoCha, 4-ClPh, 4-FPh, 3,4-diClPh, and NptGly.

[0339] In some embodiments, Xaa2 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly.

[0340] In some embodiments, Xaa2 is L. In some embodiments, Xaa2 is A. In some embodiments, Xaa2 is Cha. In some embodiments, Xaa2 is Cpa. In some embodiments, Xaa2 is (DL). In some embodiments, Xaa2 is CBA. In some embodiments, Xaa2 is MeL. In some embodiments, Xaa2 is NMeCha. In some embodiments, Xaa2 is Dcha. In some embodiments, Xaa2 is NptGly.

[0341] Xaa3 In some embodiments, Xaa3 is an α,α-disubstituted amino acid.

[0342] Xaa4 In some embodiments, Xaa4 is selected from I, Nle, MeL, AC4C, A6C, Aze, Cpa, Cha, NMeCha, Dcha, Phe(4-Cl), (β-tBu-Ala), Tle, NMeLeu, beta homoTrp, homoCha, 4-ClPh, 4-FPh, 3,4-diClPh, and NptGly.

[0343] In some embodiments, Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI). In some embodiments, Xaa4 is I. In some embodiments, Xaa4 is A. In some embodiments, Xaa4 is Nle. In some embodiments, Xaa4 is N-methyl I. In some embodiments, Xaa4 is CBA. In some embodiments, Xaa4 is (DI).

[0344] Xaa5 In some embodiments, Xaa5 is selected from R, R(Me), homoR, N-methylR, NMeArg, Lys(Me)2, Narg, Abg, Cit, Nar, Phe(4-guanidino).

[0345] In some embodiments, Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit.

[0346] In some embodiments, Xaa5 is R. In some embodiments, Xaa5 is A. In some embodiments, Xaa5 is Q. In some embodiments, Xaa5 is E. In some embodiments, Xaa5 is K. In some embodiments, Xaa5 is H. In some embodiments, Xaa5 is N-methylR. In some embodiments, Xaa5 is homoR. In some embodiments, Xaa5 is NMeArg. In some embodiments, Xaa5 is Nar. In some embodiments, Xaa5 is Cit.

[0347] Xaa6 In some embodiments, Xaa6 is an α,α-disubstituted amino acid.

[0348] Xaa7 In some embodiments, Xaa7 is selected from L, A, CBA, MeL, AC4C, A6C, Aze, Cpa, Cha, NMeCha, Dcha, Phe(4-Cl), (β-tBu-Ala), Tle, NMeLeu, beta homoTrp, homoCha, 4-ClPh, 4-FPh, 3,4-diClPh, and NptGly.

[0349] In some embodiments, Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0350] In some embodiments, Xaa7 is L. In some embodiments, Xaa7 is A. In some embodiments, Xaa7 is CBA. In some embodiments, Xaa7 is Cha. In some embodiments, Xaa7 is Cpa. In some embodiments, Xaa7 is Phe(4-Cl). In some embodiments, Xaa7 is (DL), in some embodiments, Xaa7 is α-methyl L, in some embodiments, Xaa7 is DCha, in some embodiments, Xaa7 is N-methyl Cha, in some embodiments, Xaa7 is allyl Gly, in some embodiments, Xaa7 is AC4C, in some embodiments, Xaa7 is A6C, in some embodiments, Xaa7 is Aze, in some embodiments, Xaa7 is N-MeCha, in some embodiments, Xaa7 is (β-tBu-Ala), in some embodiments, Xaa7 is Tle, in some embodiments, Xaa7 is 4-FPh, and in some embodiments, Xaa7 is 3,4-diClPh.

[0351] Xaa8 In some embodiments, Xaa8 is selected from Q, N-methyl Q, NMeGln, peptoid Q, and Ngln.

[0352] In some embodiments, Xaa8 is selected from Q and N-methylQ.

[0353] In some embodiments, Xaa8 is Q. In some embodiments, Xaa8 is N-methyl Q.

[0354] Xaa9 In some embodiments, Xaa9 is selected from an α,α-disubstituted amino acid, Q, N-methyl Q, NMeGln, peptoid Q, and Ngln.

[0355] In some embodiments, Xaa9 is selected from Q, E, N-methyl Q, N-MeGln, and peptoid Q.

[0356] In some embodiments, Xaa9 is Q. In some embodiments, Xaa9 is E. In some embodiments, Xaa9 is N-methyl Q. In some embodiments, Xaa9 is N-MeGln. In some embodiments, Xaa9 is peptoid Q.

[0357] Xaa 10 In some embodiments, Xaa 10 is selected from α,α-disubstituted amino acids, T, N-methyl T, and DThr.

[0358] In some embodiments, Xaa 10 is selected from T, N-methyl T, and DThr. 10 is T. In some embodiments, Xaa 10 is N-methyl T. In some embodiments, Xaa 10 is DThr.

[0359] Xaa 11 In some embodiments, Xaa 11 is selected from R, R(Me), homoR, N-methylR, NMeArg, Lys(Me)2, Narg, Abg, Cit, Nar, and Phe(4-guanidino).

[0360] In some embodiments, Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit.

[0361] In some embodiments, Xaa 11 is R. In some embodiments, Xaa 11 is N-methyl R. In some embodiments, Xaa 11 is E. In some embodiments, Xaa11 is K. In some embodiments, Xaa 11 is homo R. In some embodiments, Xaa 11 In some embodiments, Xaa is Nar. 11 is Cit.

[0362] Xaa 12 In some embodiments, Xaa 12 is selected from H, N-MeHis, Cys, NMeCys, homoHis, and NHis.

[0363] In some embodiments, Xaa 12 is selected from H, N-MeHis, Cys, N-MeCys, homoHis, and NHis.

[0364] In some embodiments, Xaa 12 is H. In some embodiments, Xaa 12 In some embodiments, Xaa is N-MeHis. 12 is Cys. 12 In some embodiments, Xaa is N-MeCys. 12 is homoHis. In some embodiments, Xaa 12 is NHis.

[0365] Xaa 13 In some embodiments, Xaa 13 is selected from R, R(Me), homoR, N-methylR, NMeArg, Lys(Me)2, Narg, Abg, Cit, Nar, and Phe(4-guanidino).

[0366] In some embodiments, Xaa 13 is selected from R, N-methylR, homoArg, Cit, Nar, and Phe(4-guanidino).

[0367] In some embodiments, Xaa 13is R. In some embodiments, Xaa is N-methyl R. In some embodiments, Xaa 13 is homoArg. In some embodiments, Xaa 13 In some embodiments, Xaa is Cit. 13 In some embodiments, Xaa is Nar. 13 is Phe(4-guanidino).

[0368] Xaa 14 In some embodiments, Xaa 14 is selected from α,α-disubstituted amino acids, Q, E, N-methyl E, N-methyl Q, and Ngln.

[0369] In some embodiments, Xaa 14 is selected from E, Q, N-methyl E, N-methyl Q, N-methyl D, and NMeGln.

[0370] In some embodiments, Xaa 14 is Q. In some embodiments, Xaa 14 is E. In some embodiments, Xaa 14 is N-methyl E. In some embodiments, Xaa 14 is N-methyl Q. In some embodiments, Xaa 14 is N-methyl D. In some embodiments, Xaa 14 is NMeGln.

[0371] Xaa 15 In some embodiments, Xaa 15 is selected from α,α-disubstituted amino acids, R, R(Me), homoR, N-methylR, NMeArg, Lys(Me)2, Narg, Abg, Cit, Nar, and Phe(4-guanidino).

[0372] Xaa 16 In some embodiments, Xaa 16is selected from α,α-disubstituted amino acids, S, T, and Hyp.

[0373] In some embodiments, Xaa 16 is selected from S and T. In some embodiments, Xaa 16 is S. In some embodiments, Xaa 16 is T.

[0374] In some embodiments, a polypeptide of the present disclosure comprises the following amino acid sequence: [Table 19] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0375] In some embodiments, the polypeptide has a length of 6 to 30 amino acids (eg, 6 to 12, 8 to 24, 10 to 20, or 10 to 20 amino acids).

[0376] In some embodiments, Xaa7 is CBA.

[0377] In some embodiments, Xaa7 is selected from Cha and Cpa. In some embodiments, Xaa7 is selected from α-methylL, DCha, N-methylCha, and allylGly. In some embodiments, Xaa7 is selected from AC4C, A6C, Aze, Phe(4-Cl), (β-tBu-Ala), and Tle. In some embodiments, Xaa7 is selected from Phe(4-Cl), 4-FPh, 3,4-diClPh, and Cha.

[0378] In some embodiments, Xaa7 is Cha. In some embodiments, Xaa7 is Cpa.

[0379] In some embodiments, Xaa4Xaa8Xaa5 comprises IQR.

[0380] In some embodiments, Xaa4Xaa8Xaa5 comprises I(N-methylQ)R.

[0381] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(N-methylR).

[0382] In some embodiments, Xaa4Xaa8Xaa5 comprises (CBA)QR.

[0383] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(homo R).

[0384] In some embodiments, Xaa4Xaa8Xaa5 comprises (N-methylI)QR.

[0385] In some embodiments, Xaa4Xaa8Xaa5 comprises IQQ.

[0386] In some embodiments, Xaa4Xaa8Xaa5 comprises IQE.

[0387] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(NMeArg).

[0388] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(Nar).

[0389] In some embodiments, Xaa4Xaa8Xaa5 comprises IQ(Cit).

[0390] In some embodiments, Xaa4Xaa8Xaa5 comprises IQQ, IQE, IQ(NMeArg), (Nle)QR, IQ(Nar), or IQ(Cit).

[0391] In some embodiments, the polypeptide comprises: [Table 20] During the ceremony, Xaa1 and Xaa2 are each independently selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa9 is selected from Q, E, N-methyl Q, N-MeGln, and peptoid Q; Xaa 10 is selected from T, N-methyl T, and DThr; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit.

[0392] In some embodiments, Xaa1 is L.

[0393] In some embodiments, Xaa1 is CBA.

[0394] In some embodiments, Xaa2 is L.

[0395] In some embodiments, Xaa2 is CBA.

[0396] In some embodiments, Xaa1 and Xaa2 are each L.

[0397] In some embodiments, Xaa9 is Q.

[0398] In some embodiments, Xaa9 is N-methylQ.

[0399] In some embodiments, Xaa9 is N-MeGln.

[0400] In some embodiments, Xaa1 and Xaa2 are each L and Xaa9 is Q.

[0401] In some embodiments, Xaa1 and Xaa2 are each L and Xaa9 is N-methyl Q.

[0402] In some embodiments, Xaa1 and Xaa2 are each L and Xaa9 is N-MeGln.

[0403] In some embodiments, Xaa 10 is T.

[0404] In some embodiments, Xaa 11 is selected from R, N-methylR, and homoR.

[0405] In some embodiments, Xaa 11 is R. In some embodiments, Xaa 11 is N-methyl R. In some embodiments, Xaa 11 is homo R.

[0406] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains LQTLR.

[0407] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains the L(N-methylQ)TLR.

[0408] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains L(N-methylQ)TL(homoR).

[0409] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes L(NMeGln)T(NMeLeu)R.

[0410] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes L(N-methyl Q)TL(N-methyl R).

[0411] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains the LN-methyl QTLR.

[0412] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains LETLR.

[0413] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains a (CBA) QTLR.

[0414] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains the (CBA)(N-methylQ)TLR.

[0415] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes LQT(CBA)R.

[0416] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 includes L(N-methylQ)(N-methylT)LR.

[0417] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains L(N-methylQ)T(Cha)R.

[0418] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains L(N-methylQ)T(α-methylL)R.

[0419] In some embodiments, Xaa1Xaa9Xaa 10 Xaa2Xaa 11 contains L(N-methylQ)(DThr)LR.

[0420] In some embodiments, the polypeptide comprises: [Table 21] During the ceremony, Xaa 12 is selected from H, N-MeHis, Cys, N-MeCys, homoHis, and NHis; Xaa 13 is selected from R, N-methylR, homoArg, Cit, Nar, and Phe(4-guanidino); Xaa 14 is selected from E, Q, N-methyl E, N-methyl Q, N-methyl D, and NMeGln; Xaa 15 is selected from R, homoR, and N-methylR.

[0421] In some embodiments, Xaa 12 is selected from H.

[0422] In some embodiments, Xaa 13 is R.

[0423] In some embodiments, Xaa 13 is N-methyl R.

[0424] In some embodiments, Xaa 14 is E.

[0425] In some embodiments, Xaa 14 is Q.

[0426] In some embodiments, Xaa 14 is N-methyl E.

[0427] In some embodiments, Xaa 14 is N-methyl Q.

[0428] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 includes HRER.

[0429] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 includes HRQR.

[0430] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HR(N-methylE)R.

[0431] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HR(N-methylE)R.

[0432] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HR(N-methylQ)R.

[0433] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HR(N-methylD)R.

[0434] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains H(N-methylR)QR.

[0435] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HRQ (Homo R).

[0436] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HRQ(N-methyl R).

[0437] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains H(homoArg)QR.

[0438] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 includes HRQ(NMeArg).

[0439] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains HR(NMeGln)R.

[0440] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15contains (N-MeHis)RQR.

[0441] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains (Cys)RQR.

[0442] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains (NMeCys)RQR.

[0443] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains (homoHis)RQR.

[0444] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains (NHis)RQR.

[0445] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains H(Cit)(N-methylQ)R.

[0446] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains H(Nar)(N-methylQ)R.

[0447] In some embodiments, Xaa 12 Xaa 13 Xaa 14 Xaa 15 contains H(4-guanidino-Phe)(N-methylQ)R.

[0448] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0449] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0450] In some embodiments, the polypeptide is selected from: [Table 22] TIFF2025122018000092.tif231159TIFF2025122018000093.tif233157TIFF2025122018000094.tif231161TIFF2025122018000095.tif63159In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0451] In some embodiments, the present application provides a polypeptide comprising the following amino acid sequence: [Table 23] and an amino acid sequence selected from: [Table 24] During the ceremony, Xaa3, Xaa6, Xaa 16 , and Xaa 10 are each independently an α,α-disubstituted amino acid; Xaa1 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa9 is selected from Q, N-methyl Q, E, N-MeGln, and peptoid Q; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit.

[0452] In some embodiments, the polypeptide has a length of 9 to 30 amino acids (eg, 10 to 30, 10 to 20, 12 to 30, or 12 to 20 amino acids).

[0453] In some embodiments, Xaa1 is selected from L and Cpa.

[0454] In some embodiments, Xaa1 is L. In some embodiments, Xaa1 is Cpa.

[0455] In some embodiments, Xaa9 is selected from Q and N-methylQ.

[0456] In some embodiments, Xaa9 is Q.

[0457] In some embodiments, Xaa4 is I.

[0458] In some embodiments, Xaa8 is Q.

[0459] In some embodiments, Xaa5 is R.

[0460] In some embodiments, Xaa1Xaa9 is LQ.

[0461] In some embodiments, Xaa1Xaa9 is L(N-methylQ).

[0462] In some embodiments, Xaa1Xaa9 is (Cpa)(N-methylQ).

[0463] In some embodiments, Xaa4Xaa8Xaa5 is IQR.

[0464] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10and Xaa3Xaa4Xaa8Xaa5Xaa6, as well as at least one R (e.g., one R, two R, or three R).

[0465] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus at least one L (e.g., 1 L, 2 L, or 3 L).

[0466] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, and at least one amino acid selected from CBA, Cpa, and Cha. In some embodiments, the polypeptide comprises CBA. In some embodiments, the polypeptide comprises Cpa. In some embodiments, the polypeptide comprises Cha.

[0467] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus at least one (2-Nal) (e.g., one, two, or three 2-Nal).

[0468] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, as well as at least one (β-Ala). In some embodiments, the polypeptide comprises (β-Ala)(β-Ala).

[0469] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, as well as HRQR. In some embodiments, the polypeptide comprises the sequence Xaa16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus HRER.

[0470] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus LR.

[0471] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus (Cpa)R.

[0472] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6 plus (Cha)R.

[0473] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus (CBA)(2-Nal).

[0474] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus (Cpa)(2-Nal).

[0475] In some embodiments, the polypeptide has the sequence Xaa 16 Xaa1Xaa9Xaa 10 and Xaa3Xaa4Xaa8Xaa5Xaa6, plus (Cha)(2-Nal).

[0476] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0477] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0478] In some embodiments, Xaa 16 and Xaa 10 In some aspects of these embodiments, Xaa 16 and Xaa 10 and Xaa are each (S)-2-(4'-pentenyl)alanine. 16 and Xaa 10 and Xaa are each (R)-2-(4'-pentenyl)alanine. 16 and Xaa 10 and Xaa are each (S)-2-(7'-octenyl)alanine. 16 and Xaa 10 are each (R)-2-(7'-octenyl)alanine.

[0479] In some embodiments, Xaa 16 and Xaa 10 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa 16 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa 16 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (R)-2-(7'-octenyl)alanine, while Xaa 10In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 16 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 16 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 16 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (R)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 16 is (R)-2-(7'-octenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine.

[0480] In some embodiments, Xaa3, Xaa6, and Xaa 16 and Xaa 10In some aspects of these embodiments, Xaa3, Xaa6, Xaa 16 , and Xaa 10 In other aspects of these embodiments, Xaa3, Xaa6, Xaa are each (S)-2-(4'-pentenyl)alanine. 16 , and Xaa 10 In still other aspects of these embodiments, Xaa3, Xaa6, Xaa are each (R)-2-(4'-pentenyl)alanine. 16 , and Xaa 10 In still other aspects of these embodiments, Xaa3, Xaa6, Xaa are each (S)-2-(7'-octenyl)alanine. 16 , and Xaa 10 are each (R)-2-(7'-octenyl)alanine.

[0481] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine, and Xaa 16 and Xaa 10 are each (R)-2-(4'-pentenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine and Xaa 16 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine, while Xaa 16 and Xaa 10 are each (R)-2-(7'-octenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine, and Xaa 16 and Xaa 10 In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine, and Xaa 16 and Xaa 10are each (S)-2-(7'-octenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine and Xaa 16 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine, while Xaa 16 and Xaa 10 are each (R)-2-(7'-octenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine, while Xaa 16 and Xaa 10 are each (R)-2-(4'-pentenyl)alanine.

[0482] In some embodiments, the polypeptide has the following amino acid sequence: RXaa 16 L(N-methylQ)Xaa 10 LRXaa3IQRXaa6(CBA)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 88), In the formula, Xaa3, Xaa6, Xaa 16 , and Xaa 10 are each (R)-2-(4'-pentenyl)alanine.

[0483] In some embodiments, the polypeptide has the following amino acid sequence: RXaa 16 (Cpa)(N-methylQ)Xaa 10 (Cpa)RXaa3IQRXaa6(Cpa)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 89), In the formula, Xaa3, Xaa6, Xaa 16 , and Xaa 10 are each (R)-2-(4'-pentenyl)alanine.

[0484] In some embodiments, the present application provides the following amino acid sequence: [Table 25] and an amino acid sequence selected from: [Table 26] During the ceremony, Xaa3, Xaa6, Xaa9, and Xaa 14 are each independently an α,α-disubstituted amino acid; Xaa 15 is selected from R, homoR, and N-methylR; Xaa 16 is selected from S and T, Xaa1 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit The present invention provides a polypeptide having the formula:

[0485] In some embodiments, the polypeptide has a length of 9 to 30 amino acids (eg, 10 to 30, 10 to 20, 12 to 20, or 12 to 30 amino acids).

[0486] In some embodiments, Xaa 15 is R.

[0487] In some embodiments, Xaa 16 is S.

[0488] In some embodiments, Xaa1 is L.

[0489] In some embodiments, Xaa 15 Xaa 16 Xaa1 contains the RSL.

[0490] In some embodiments, Xaa4 is I.

[0491] In some embodiments, Xaa8 is Q.

[0492] In some embodiments, Xaa5 is R.

[0493] In some embodiments, Xaa4Xaa8Xaa5 comprises IQR.

[0494] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one R.

[0495] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (2-Nal).

[0496] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains HR.

[0497] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa 15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains a TLR.

[0498] In some embodiments, the polypeptide has the amino acid sequence Xaa 14 Xaa15 Xaa 16 In addition to Xaa1Xaa9 and the amino acid sequence Xaa3Xaa4Xaa8Xaa5Xaa6, it contains (CBA)(2-Nal) or (4-ClPh)(2-Nal).

[0499] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0500] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0501] In some embodiments, Xaa9 and Xaa 14 In some aspects of these embodiments, Xaa9 and Xaa 14 In other aspects of these embodiments, Xaa9 and Xaa 14 In still other aspects of these embodiments, Xaa9 and Xaa 14 In still other aspects of these embodiments, Xaa9 and Xaa 14 are each (R)-2-(7'-octenyl)alanine.

[0502] In some embodiments, Xaa9 and Xaa 14 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa 16 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 In other aspects of these embodiments, Xaa9 is (R)-2-(4'-pentenyl)alanine, while Xaa 14 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa9 is (S)-2-(7'-octenyl)alanine, while Xaa 14 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 14 is (R)-2-(7'-octenyl)alanine while Xaa9 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa9 is (S)-2-(4'-pentenyl)alanine while Xaa 14is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa9 is (R)-2-(4'-pentenyl)alanine, while Xaa 14 In still other aspects of these embodiments, Xaa9 is (S)-2-(4'-pentenyl)alanine, while Xaa 14 In still other aspects of these embodiments, Xaa9 is (R)-2-(4'-pentenyl)alanine, while Xaa 14 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa9 is (S)-2-(7'-octenyl)alanine, while Xaa 14 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa9 is (R)-2-(7'-octenyl)alanine, while Xaa 14 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa9 is (S)-2-(7'-octenyl)alanine, while Xaa 14 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa9 is (R)-2-(7'-octenyl)alanine, while Xaa 14 is (S)-2-(4'-pentenyl)alanine.

[0503] In some embodiments, Xaa3, Xaa6, Xaa9, and Xaa 14 In some aspects of these embodiments, Xaa3, Xaa6, Xaa9, and Xaa 14 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3, Xaa6, Xaa9, and Xaa 14 In still other aspects of these embodiments, Xaa3, Xaa6, Xaa9, and Xaa are each (R)-2-(4'-pentenyl)alanine. 14In still other aspects of these embodiments, Xaa3, Xaa6, Xaa9, and Xaa are each (S)-2-(7'-octenyl)alanine. 14 are each (R)-2-(7'-octenyl)alanine.

[0504] In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine, and Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine, and Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In some embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine, while Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine, and Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine, and Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine, and Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine, while Xaa9 and Xaa 14 In some embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine, while Xaa9 and Xaa 14 are each (R)-2-(4'-pentenyl)alanine.

[0505] In some embodiments, the polypeptide has the following amino acid sequence: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 94), In the formula, Xaa3, Xaa6, Xaa9, and Xaa 14 are each (S)-2-(4'-pentenyl)alanine.

[0506] In some embodiments, the polypeptide has the following amino acid sequence: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 95), In the formula, Xaa3, Xaa6, Xaa9, and Xaa 14 are (S)-2-(4'-pentenyl)alanine, In this formula, the N-terminus is modified with palmitoyl-PEG4.

[0507] In some embodiments, the polypeptide has the following amino acid sequence: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-ClPh)(2-Nal) (SEQ ID NO: 96), In the formula, Xaa3, Xaa6, Xaa9, and Xaa 14 are (S)-2-(4'-pentenyl)alanine, In this formula, the N-terminus is modified with palmitoyl-PEG4.

[0508] In some embodiments, the polypeptide has the following amino acid sequence: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-Cl-Ph)(2-Nal) (SEQ ID NO: 198), In the formula, Xaa3, Xaa6, Xaa9, and Xaa 14 are each (S)-2-(4'-pentenyl)alanine.

[0509] In some embodiments, the present application provides a polypeptide having the following amino acid sequence: [Table 27] During the ceremony, Xaa 10 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa2 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit; Xaa3 is selected from D and Nle; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit.

[0510] In some embodiments, the polypeptide has a length of 8 to 30 amino acids (eg, 10 to 30, 10 to 20, 12 to 30, and 12 to 20 amino acids).

[0511] In some embodiments, Xaa2 is L.

[0512] In some embodiments, Xaa 11 is R.

[0513] In some embodiments, Xaa3 is D.

[0514] In some embodiments, Xaa4 is I.

[0515] In some embodiments, Xaa8 is Q.

[0516] In some embodiments, Xaa5 is R.

[0517] In some embodiments, Xaa2Xaa 11 Xaa3Xaa4Xaa8Xaa5 contains LRDIQR.

[0518] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one L.

[0519] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (2-Nal).

[0520] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (β-Ala).

[0521] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains LQ.

[0522] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains L(2-Nal).

[0523] In some embodiments, the polypeptide has the amino acid sequence Xaa 10 Xaa2Xaa 11 In addition to Xaa3Xaa4Xaa8Xaa5Xaa6, it contains at least one (β-Ala).

[0524] In some embodiments, the polypeptide has the amino acid sequence Xaa10 Xaa2Xaa 11 Xaa3Xaa4Xaa8Xaa5Xaa6 plus HRERS or HRQRS.

[0525] In some embodiments, Xaa 10 and Xaa6 are the same. In some aspects of these embodiments, Xaa 10 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. 10 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. 10 and Xaa6 are each (S)-2-(7'-octenyl)alanine. 10 and Xaa6 is each (R)-2-(7'-octenyl)alanine.

[0526] In some embodiments, Xaa 10 and Xaa6 are different α,α-disubstituted amino acids. 10 is (S)-2-(4'-pentenyl)alanine, while Xaa6 is (R)-2-(4'-pentenyl)alanine. 10 is (R)-2-(4'-pentenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine. 10 is (S)-2-(7'-octenyl)alanine, while Xaa6 is (R)-2-(7'-octenyl)alanine. 10 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(7'-octenyl)alanine. 10is (S)-2-(4'-pentenyl)alanine, while Xaa6 is (S)-2-(7'-octenyl)alanine. 10 is (R)-2-(4'-pentenyl)alanine, while Xaa6 is (R)-2-(7'-octenyl)alanine. 10 is (S)-2-(4'-pentenyl)alanine, while Xaa6 is (R)-2-(7'-octenyl)alanine. 10 is (R)-2-(4'-pentenyl)alanine, while Xaa6 is (S)-2-(7'-octenyl)alanine. 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine, while Xaa is (S)-2-(4'-pentenyl)alanine. 10 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (R)-2-(4'-pentenyl)alanine. 10 is (S)-2-(7'-octenyl)alanine, while Xaa6 is (R)-2-(4'-pentenyl)alanine. 10 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0527] In some embodiments, the polypeptide has the following amino acid sequence: LQXaa 10 LRDIQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 97) In the formula, Xaa6 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine.

[0528] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence SEQ ID NO:1 shown below: [Table 28]

[0529] With reference to a polypeptide comprising the amino acid sequence SEQ ID NO: 1, Xaa1 and Xaa2 are each independently selected from L, A, Cha, Cpa, CBA, (DL), MeL, NMeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0530] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0531] In some embodiments, the polypeptide comprising the amino acid sequence SEQ ID NO:1 has a length of 13 to 30 amino acids (eg, 13 to 22, 14 to 30, 16 to 30, 18 to 30, 14 to 22, or 14 to 20 amino acids).

[0532] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence SEQ ID NO: 15, as shown below: [Table 29]

[0533] With reference to a polypeptide comprising the amino acid sequence SEQ ID NO: 15, Xaa1 and Xaa2 are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0534] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0535] In some embodiments, a polypeptide comprising the amino acid sequence SEQ ID NO: 15 also comprises at least one A (e.g., two or more A). In some embodiments, a polypeptide comprising the amino acid sequence SEQ ID NO: 15 has a length of 12 to 30 amino acids (e.g., 12 to 22, 12 to 22, 14 to 22, 12 to 20, or 12 to 18 amino acids).

[0536] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence SEQ ID NO:30, as shown below: [Table 30]

[0537] With reference to a polypeptide comprising the amino acid sequence SEQ ID NO: 30, Xaa1 and Xaa2 are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0538] In some embodiments, Xaa1 and Xaa2 are each independently L or A; Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is I or A, Xaa5 is R or A, Xaa7 is L, A, or CBA.

[0539] In some embodiments, the polypeptide comprising the amino acid sequence SEQ ID NO:30 also comprises at least one A. In some embodiments, the polypeptide comprising the amino acid sequence SEQ ID NO:30 has a length of 12 to 30 amino acids.

[0540] Specific embodiments of polypeptides comprising the amino acid sequences SEQ ID NO:1, SEQ ID NO:15, and SEQ ID NO:30 are described below.

[0541] In some embodiments, Xaa1 is L. In some embodiments, Xaa1 is A. In some embodiments, Xaa2 is L. In some embodiments, Xaa2 is A. In some embodiments, Xaa1 and Xaa2 are each L. In some embodiments, Xaa1 and Xaa2 are each A. In some embodiments, Xaa1 is L and Xaa2 is A. In some embodiments, Xaa1 is A and Xaa2 is L. In some embodiments, Xaa1 and Xaa2 are each Cha. In some embodiments, Xaa1 and Xaa2 are each Cpa.

[0542] In some embodiments, Xaa4 is A. In some embodiments, Xaa4 is I. In some embodiments, Xaa2 and Xaa4 are each A. In some embodiments, Xaa2 is L and Xaa4 is A. In some embodiments, Xaa2 is L and Xaa4 is I. In some embodiments, Xaa2 is A and Xaa4 is I.

[0543] In some embodiments, Xaa5 is R. In some embodiments, Xaa5 is A. In some embodiments, Xaa4 is I and Xaa5 is R. In some embodiments, Xaa4 is I and Xaa5 is A. In some embodiments, Xaa4 is A and Xaa5 is R. In some embodiments, Xaa4 and Xaa5 are both A.

[0544] In some embodiments, Xaa7 is L. In some embodiments, Xaa7 is A. In some embodiments, Xaa7 is CBA. In some embodiments, Xaa7 is selected from Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0545] In some embodiments, Xaa5 is R and Xaa7 is L. In some embodiments, Xaa5 is R and Xaa7 is A. In some embodiments, Xaa5 is A and Xaa7 is L. In some embodiments, Xaa5 is R and Xaa7 is CBA. In some embodiments, Xaa5 is A and Xaa7 is CBA. In some embodiments, Xaa4 is I and Xaa7 is A.

[0546] In some embodiments, at least one of Xaa1, Xaa2, Xaa4, Xaa5, and Xaa7 is A. In some aspects of these embodiments, at least two of Xaa1, Xaa2, Xaa4, Xaa5, and Xaa7 are A. In other aspects of these embodiments, at least Xaa1 is A. In still other aspects of these embodiments, at least Xaa2 is A. In still other aspects of these embodiments, at least Xaa4 is A. In still other aspects of these embodiments, at least Xaa5 is A. In still other aspects of these embodiments, at least Xaa7 is A.

[0547] In some embodiments, Xaa1 and Xaa2 are each A, X4 is I, X5 is R, and X7 is L. In some embodiments, Xaa1 is L, Xaa2 is A, X4 is A, X5 is R, and X7 is L. In some embodiments, Xaa1 and Xaa2 are each L, X4 is I, X5 is R, and X7 is L. In some embodiments, Xaa1 and Xaa2 are each L, X4 is I, X5 is A, and X7 is L. In some embodiments, Xaa1 and Xaa2 are each L, X4 is I, X5 is R, and X7 is CBA. In some embodiments, Xaa1 and Xaa2 are each L, X4 is I, X5 is A, and X7 is CBA.

[0548] In some embodiments, the polypeptide of the present application comprises the amino acid sequence SEQ ID NO:1, SEQ ID NO:15, or SEQ ID NO:30, and additionally at least one amino acid selected from Q, L, E, H, and R. In some embodiments, the polypeptide of the present application comprises the amino acid sequence SEQ ID NO:1, SEQ ID NO:15, or SEQ ID NO:30, and additionally at least one amino acid selected from Q, L, E, H, R, and S. In some embodiments, the polypeptide of the present application comprises the amino acid sequence SEQ ID NO:1, SEQ ID NO:15, or SEQ ID NO:30, and additionally at least one amino acid selected from P, D, and β-Ala. In some embodiments, the polypeptide comprises at least two Qs. In some embodiments, the polypeptide comprises at least three Qs. In some embodiments, the polypeptide comprises at least two Ls. In some embodiments, the polypeptide comprises at least three Ls. In some embodiments, the polypeptide comprises at least four Ls. In some embodiments, the polypeptide comprises at least two Rs. In some embodiments, the polypeptide comprises at least three Rs. In some embodiments, the polypeptide comprises at least four Rs. In some embodiments, the polypeptide comprises at least two Es. In some embodiments, the polypeptide comprises at least one H and at least one I. In some embodiments, the polypeptide of the present application comprises any of the amino acid sequences listed in Table 3, or a combination thereof, in addition to the amino acid sequence SEQ ID NO:1, SEQ ID NO:15, or SEQ ID NO:30.

[0549] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0550] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0551] In some embodiments, the amino acid sequence SEQ ID NO:1 is selected from the following: SLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 2), and SLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 3).

[0552] In some embodiments, the amino acid sequence SEQ ID NO:15 is selected from the following: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 16), LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 17), LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 18), LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 19), and LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 20).

[0553] In some embodiments, the amino acid sequence SEQ ID NO:30 is: LQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 31).

[0554] In some embodiments, the polypeptide consists of the following amino acid sequence: RSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 4), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0555] In some embodiments, the polypeptide consists of the following amino acid sequence: RERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 5), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0556] In some embodiments, the polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 6), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0557] In some embodiments, the polypeptide consists of the following amino acid sequence: EHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 7), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0558] In some embodiments, the polypeptide consists of the following amino acid sequence: QLEHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 8), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0559] In some embodiments, the polypeptide consists of the following amino acid sequence: EHRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 9), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0560] In some embodiments, the polypeptide consists of the following amino acid sequence: QERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 10), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0561] In some embodiments, the polypeptide consists of the following amino acid sequence: HQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 11), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0562] In some embodiments, the polypeptide consists of the following amino acid sequence: EHQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 12), wherein Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0563] In some embodiments, the polypeptide consists of the following amino acid sequence: RERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 13), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0564] In some embodiments, the polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 14), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0565] In some embodiments, the polypeptide consists of the following amino acid sequence: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 21), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0566] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 22), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0567] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 23), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0568] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 24), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0569] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 25), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0570] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 26), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0571] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal)AA (SEQ ID NO: 27), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0572] In some embodiments, the polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 28), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0573] In some embodiments, the polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 29), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0574] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 75), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0575] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 76), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0576] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6A(2-Nal) (SEQ ID NO: 77), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0577] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)PD (SEQ ID NO: 32), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0578] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)P (SEQ ID NO: 33), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0579] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0580] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34a), wherein Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine; wherein the C-terminus of SEQ ID NO: 34a is modified with GRKKRRQRRRPQK(PEG4-palmitoyl)NH2.

[0581] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence SEQ ID NO:35, as shown below: [Table 31]

[0582] With reference to a polypeptide comprising the amino acid sequence SEQ ID NO: 35, Xaa1 and Xaa3 are each independently an α,α-disubstituted amino acid; Xaa2 is selected from L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly.

[0583] In some embodiments, Xaa1 and Xaa3 are each independently an α,α-disubstituted amino acid; Xaa2 is L or A.

[0584] In some embodiments, a polypeptide comprising the amino acid sequence SEQ ID NO: 35 also comprises at least one 2-Nal (e.g., two or more 2-Nal). In some embodiments, the polypeptide has a length of 6 to 30 amino acids (e.g., 6 to 22, 7 to 22, 9 to 22, 12 to 22, 6 to 20, 7 to 20, 9 to 20, or 12 to 20 amino acids).

[0585] Specific embodiments of polypeptides comprising the amino acid sequence SEQ ID NO:35 are described below.

[0586] In some embodiments, Xaa2 is L. In some embodiments, Xaa2 is A.

[0587] In some embodiments, Xaa1 and Xaa3 are the same. In some aspects of these embodiments, Xaa1 and Xaa3 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa1 and Xaa3 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 and Xaa3 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 and Xaa3 are each (R)-2-(7'-octenyl)alanine.

[0588] In some embodiments, Xaa1 and Xaa3 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa3 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa3 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa3 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine while Xaa3 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa3 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa3 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa3 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa3 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa3 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine while Xaa3 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa3 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine, while Xaa3 is (S)-2-(4'-pentenyl)alanine.

[0589] In some embodiments, a polypeptide comprises the amino acid sequence SEQ ID NO:35 plus at least one amino acid selected from Q, L, E, H, I, S, M, and R. In some embodiments, a polypeptide comprises the amino acid sequence SEQ ID NO:35 plus at least one amino acid selected from H, S, I, and M. In some embodiments, a polypeptide comprises at least two Qs. In some embodiments, a polypeptide comprises at least two Ls. In some embodiments, a polypeptide comprises at least three Ls. In some embodiments, a polypeptide comprises at least four Ls. In some embodiments, a polypeptide comprises at least two Rs. In some embodiments, a polypeptide comprises at least three Rs. In some embodiments, a polypeptide comprises at least four Rs. In some embodiments, a polypeptide comprises at least two Es. In some embodiments, a polypeptide comprises at least one H and at least one I. In some embodiments, a polypeptide of the present application comprises, in addition to the amino acids of SEQ ID NO:35, any of the amino acid sequences listed in Table 3, or a combination thereof. In some embodiments, a polypeptide comprises, in addition to the amino acid sequence SEQ ID NO:35, the amino acid sequence ML(2-Nal). In some embodiments, a polypeptide comprising the amino acid sequence SEQ ID NO: 35 also comprises the following amino acid sequences: IQR, RERSL, QLEH, and ML(2-Nal). In some embodiments, a polypeptide comprising the amino acid sequence SEQ ID NO: 35 also comprises the following amino acid sequence: (2-Abu)L(2-Nal)(β-Ala)(β-Ala).

[0590] In some embodiments, the polypeptide consists of the following amino acid sequence: QLEHRERSLXaa1TLRXaa3IQRML(2-Nal) (SEQ ID NO: 36), In the formula, Xaa1 and Xaa3 are each (S)-2-(4'-pentenyl)alanine.

[0591] In some embodiments, the polypeptide consists of the following amino acid sequence: QLEHRERSLXaa1TLRXaa3IQR(2-Abu)L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 78), wherein Xaa1 and Xaa3 are each (S)-2-(4'-pentenyl)alanine.

[0592] In some embodiments, the polypeptide of the present disclosure has an amino acid sequence selected from the following: Xaa1SLQXaa2 (SEQ ID NO: 37a) and Xaa1S(Cha)(N-methylQ)Xaa2 (SEQ ID NO: 37b), and an amino acid sequence selected from the following: Xaa3IQRXaa4 (SEQ ID NO: 38a) and Xaa3IQQXaa4 (SEQ ID NO: 38b), wherein Xaa1, Xaa2, Xaa3, and Xaa4 are each independently an α,α-disubstituted amino acid.

[0593] In some embodiments, a polypeptide of the present disclosure comprises the amino acid sequence shown below, SEQ ID NO:37 and SEQ ID NO:38: [Table 32]

[0594] With reference to the polypeptide comprising the amino acid sequences SEQ ID NO:37 and SEQ ID NO:38, Xaa1, Xaa2, Xaa3, and Xaa4 are each independently an α,α-disubstituted amino acid. In some embodiments, the polypeptide has a length of 10 to 30 amino acids (e.g., 10 to 20, 10 to 22, 12 to 22, 12 to 20, 14 to 22, or 14 to 20 amino acids).

[0595] Specific embodiments of polypeptides comprising the amino acid sequences SEQ ID NO:37 and SEQ ID NO:38 are described below.

[0596] In some embodiments, the polypeptide comprises at least one 2-Nal.

[0597] In some embodiments, the polypeptide comprises at least one CBA.

[0598] In some embodiments, the polypeptides of the present application comprise any of the amino acid sequences listed in Table 3, or a combination thereof, in addition to the amino acid sequences SEQ ID NOs:37 and 38. In some embodiments, the polypeptides comprise at least one amino acid selected from L, E, and R in addition to the amino acid sequences SEQ ID NOs:37 and 38. In some embodiments, the polypeptides comprise at least two Ls. In some embodiments, the polypeptides comprise at least three Ls. In some embodiments, the polypeptides comprise at least two Rs. In some embodiments, the polypeptides of the present application comprise any of the amino acid sequences listed in Table 3, or a combination thereof, in addition to the amino acid sequences SEQ ID NOs:37 and 38. In some embodiments, the polypeptides comprising the amino acid sequences SEQ ID NOs:37 and 38 also comprise at least one amino acid selected from RE, LR, and L(2-Nal). In some embodiments, the polypeptides comprise the following amino acid sequence: Xaa2LRXaa3. In some embodiments, the polypeptides comprise at least one β-Ala. In some embodiments, the polypeptides comprise the amino acid sequence (β-Ala)(β-Ala). In some embodiments, the polypeptide comprises at least one amino acid selected from L, E, R, H, Q, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, homoHis, NHis, homoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylic Gly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu. In some embodiments, the polypeptide comprises at least one amino acid selected from L, E, R, H, Q, N-methyl E, CBA, N-methyl Q, Cha, and N-methyl R.

[0599] In some embodiments, the polypeptide comprises the amino acid sequence (CBA)(2-Nal), hi some embodiments, the polypeptide comprises an amino acid sequence selected from HRE, HR(N-methyl E), HR(N-methyl Q), HRQ, LR, L(N-methyl R), (Cha)R, L(2-Nal), and (CBA)(2-Nal).

[0600] In some embodiments, Xaa1 and Xaa2 are the same. In some aspects of these embodiments, Xaa1 and Xaa2 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa1 and Xaa2 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 and Xaa2 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 and Xaa2 are each (R)-2-(7'-octenyl)alanine.

[0601] In some embodiments, Xaa1 and Xaa2 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa2 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa2 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa2 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine while Xaa2 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa2 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa2 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(4'-pentenyl)alanine while Xaa2 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(4'-pentenyl)alanine while Xaa2 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa2 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine while Xaa2 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1 is (S)-2-(7'-octenyl)alanine while Xaa2 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa1 is (R)-2-(7'-octenyl)alanine, while Xaa2 is (S)-2-(4'-pentenyl)alanine.

[0602] In some embodiments, Xaa3 and Xaa4 are the same. In some aspects of these embodiments, Xaa3 and Xaa4 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa4 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa4 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa4 are each (R)-2-(7'-octenyl)alanine.

[0603] In some embodiments, Xaa3 and Xaa4 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa4 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa4 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa4 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa4 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa4 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa4 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa4 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa4 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa4 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa4 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa4 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa4 is (S)-2-(4'-pentenyl)alanine.

[0604] In some embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are all identical. In some aspects of these embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa1, Xaa2, Xaa3, and Xaa4 are each (R)-2-(7'-octenyl)alanine.

[0605] In some embodiments, Xaa1 and Xaa2 are each (S)-2-(4'-pentenyl)alanine, and Xaa3 and Xaa4 are each (R)-2-(4'-pentenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (R)-2-(4'-pentenyl)alanine, and Xaa3 and Xaa4 are each (S)-2-(4'-pentenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (S)-2-(7'-octenyl)alanine, while Xaa3 and Xaa4 are each (R)-2-(7'-octenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (R)-2-(7'-octenyl)alanine, while Xaa3 and Xaa4 are each (S)-2-(7'-octenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (S)-2-(4'-pentenyl)alanine, and Xaa3 and Xaa4 are each (S)-2-(7'-octenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (S)-2-(7'-octenyl)alanine, and Xaa3 and Xaa4 are each (S)-2-(4'-pentenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (R)-2-(4'-pentenyl)alanine, while Xaa3 and Xaa4 are each (R)-2-(7'-octenyl)alanine. In some embodiments, Xaa1 and Xaa2 are each (R)-2-(7'-octenyl)alanine, while Xaa3 and Xaa4 are each (R)-2-(4'-pentenyl)alanine.

[0606] In some embodiments, the polypeptide consists of the following amino acid sequence: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal) (SEQ ID NO: 39), In the formula, Xaa1, Xaa2, Xaa3, and Xaa4 are each (S)-2-(4'-pentenyl)alanine.

[0607] In some embodiments, the polypeptide consists of the following amino acid sequence: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 40), In the formula, Xaa1, Xaa2, Xaa3, and Xaa4 are each (S)-2-(4'-pentenyl)alanine.

[0608] In some embodiments, the polypeptide is selected from any one of the following polypeptides: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal) (SEQ ID NO: 39), REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 40), EXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 79), HREXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 82), HREXaa1SLQXaa2LRXaa3IQQXaa4(CBA)(2-Nal) (SEQ ID NO: 83), HR(N-methyl E)Xaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 84), HREXaa1SLQXaa2L(N-methylR)Xaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 85), HR(N-methylQ)Xaa1S(Cha)(N-methylQ)Xaa2(Cha)RXaa3IQRXaa4(Cha)(2-Nal) (SEQ ID NO: 86), and HRQXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 87), In the formula, Xaa1, Xaa2, Xaa3, and Xaa4 are each (S)-2-(4'-pentenyl)alanine.

[0609] In some embodiments, the present application provides the following amino acid sequence SEQ ID NO:37: [Table 33] During the ceremony, Xaa 15 and Xaa 10 are each independently an α,α-disubstituted amino acid.

[0610] In some embodiments, the polypeptide comprises at least two Nle.

[0611] In some embodiments, the polypeptide has a length of 7 to 30 amino acids (eg, 7 to 20, 8 to 25, 10 to 30, 12 to 30, or 9 to 24 amino acids).

[0612] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO:37, as well as at least one amino acid selected from L, E, R, H, S, Q, I, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, HomoHis, NHis, HomoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylic Gly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

[0613] In some embodiments, the polypeptide comprises the amino acid sequence SEQ ID NO: 37 plus at least one amino acid selected from H, R, E, S, L, Q, I, CBA, and (2-Nal).

[0614] In some embodiments, the polypeptide comprises an IQR.

[0615] In some embodiments, the polypeptide comprises (Nle)IQR(Nle).

[0616] In some embodiments, the polypeptide comprises an HRE and a LR.

[0617] In some embodiments, the polypeptide comprises L(2-Nal).

[0618] In some embodiments, the polypeptide comprises (CBA)(2-Nal).

[0619] In some embodiments, Xaa 15 and Xaa 10 In some aspects of these embodiments, Xaa 15 and Xaa 10 and Xaa are each (S)-2-(4'-pentenyl)alanine. 15 and Xaa 10 and Xaa are each (R)-2-(4'-pentenyl)alanine. 15 and Xaa 10 and Xaa are each (S)-2-(7'-octenyl)alanine. 15 and Xaa 10 are each (R)-2-(7'-octenyl)alanine.

[0620] In some embodiments, Xaa 15 and Xaa 10 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa 15 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa 15 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 15is (R)-2-(7'-octenyl)alanine, while Xaa 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 15 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 15 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (S)-2-(4'-pentenyl)alanine, while Xaa 10 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (R)-2-(4'-pentenyl)alanine, while Xaa 10 In still other aspects of these embodiments, Xaa is (S)-2-(7'-octenyl)alanine. 15 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (R)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (S)-2-(7'-octenyl)alanine, while Xaa 10 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa 15 is (R)-2-(7'-octenyl)alanine, while Xaa 10 is (S)-2-(4'-pentenyl)alanine.

[0621] In some embodiments, the polypeptide consists of the following amino acid sequence: HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)L(2-Nal) (SEQ ID NO: 80), In the formula, Xaa 15 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine.

[0622] In some embodiments, the polypeptide consists of the following amino acid sequence: HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)(CBA)(2-Nal) (SEQ ID NO: 81), In the formula, Xaa 15 and Xaa 10 are each (S)-2-(4'-pentenyl)alanine.

[0623] In some embodiments, the disclosure provides a polypeptide having the following amino acid sequence: [Table 34] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; Xaa 11 is selected from R, N-methylR, E, K, homoR, Nar, and Cit.

[0624] In some embodiments, Xaa 11 and at least one of Xaa5 contains E or K.

[0625] In some embodiments, the polypeptide has a length of 8 to 30 amino acids (eg, 10 to 25, 12 to 24, 9 to 30, 10 to 30, or 12 to 30 amino acids).

[0626] In some embodiments, Xaa 11 is E. In some embodiments, Xaa 11 is R. In some embodiments, Xaa 11 In some embodiments, Xaa5 is K. In some embodiments, Xaa5 is R. In some embodiments, Xaa5 is E. In some embodiments, Xaa5 is K. In some embodiments, Xaa 11 is E and Xaa5 is R. In some embodiments, Xaa 11 is R and Xaa5 is E. In some embodiments, Xaa 11 is K and Xaa5 is R. In some embodiments, Xaa 11 is R and Xaa5 is K.

[0627] In some embodiments, the polypeptide comprises L(2-Nal). In some embodiments, the polypeptide comprises IQR, IQE, or IQK. In some embodiments, the polypeptide comprises LQTLE, LQTLR, or LQRLK.

[0628] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0629] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0630] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLEXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 98), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0631] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQEXaa6L(2-Nal) (SEQ ID NO: 99), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0632] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLKXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 100), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0633] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQKXaa6L(2-Nal) (SEQ ID NO: 101), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0634] In some embodiments, the application provides a polypeptide having the following sequence: [Table 35] During the ceremony, Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid; Xaa4 is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa8 is selected from Q and N-methyl Q; Xaa5 is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa7 is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

[0635] In some embodiments, the polypeptide comprises at least one of (DI) or (DL).

[0636] In some embodiments, the polypeptide has a length of 7 to 30 amino acids (eg, 7 to 12, 8 to 30, 8 to 25, 10 to 30, 12 to 24, or 12 to 20 amino acids).

[0637] In some embodiments, Xaa4 is (DI). In some embodiments, Xaa4 is (DL).

[0638] In some embodiments, Xaa7 is (DL). In some embodiments, Xaa7 is (DI).

[0639] In some embodiments, the polypeptide comprises a (DL)QTIR.

[0640] In some embodiments, the polypeptide comprises LQT(DL)R.

[0641] In some embodiments, Xaa3 and Xaa6 are the same. In some aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 and Xaa6 are each (R)-2-(7'-octenyl)alanine.

[0642] In some embodiments, Xaa3 and Xaa6 are different α,α-disubstituted amino acids. In some aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(4'-pentenyl)alanine while Xaa6 is (R)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(4'-pentenyl)alanine while Xaa6 is (S)-2-(7'-octenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (S)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine. In still other aspects of these embodiments, Xaa3 is (S)-2-(7'-octenyl)alanine while Xaa6 is (R)-2-(4'-pentenyl)alanine.In still other aspects of these embodiments, Xaa3 is (R)-2-(7'-octenyl)alanine, while Xaa6 is (S)-2-(4'-pentenyl)alanine.

[0643] In some embodiments, the polypeptide consists of the following amino acid sequence: (DL)QTIRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 102), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0644] In some embodiments, the polypeptide consists of the following amino acid sequence: LQT(DL)RXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 103), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0645] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3(DI)QRXaa6L(2-Nal) (SEQ ID NO: 104), In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0646] In some embodiments, the polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6(DL)(2-Nal) (SEQ ID NO: 105). In the formula, Xaa3 and Xaa6 are each (S)-2-(4'-pentenyl)alanine.

[0647] Stapled polypeptide derived from the HD2 domain of BCL9 Stabilizing peptides have been shown to offer benefits such as increased helical content, proteolytic stability, and increased binding affinity to target receptors (see Kim 2011). In particular, α-helical domains are known to be amenable to stabilization.

[0648] In some embodiments, the polypeptides disclosed herein include polypeptides that undergo a reaction (e.g., a metathesis reaction) to form one or more hydrocarbon linkers, and thus include polypeptides that contain one or more hydrocarbon linkers. The hydrocarbon linkers can impose structural constraints on the α-helix of the polypeptide (e.g., any variant of the HD2 domain of BCL9 described herein). In one embodiment, the α-helix of the polypeptide is stabilized by having one or more hydrocarbon linkers between the amino acids of the polypeptide.

[0649] The hydrocarbon crosslinker can extend the length of one or more α-helical turns, since it is generally understood that an α-helical turn contains approximately 3-4 amino acids. Thus, any two amino acids chemically linked by the hydrocarbon linker are at positions i and i+4 relative to each other.

[0650] In some embodiments, the hydrocarbon crosslinkers disclosed herein are produced by connecting two α,α-disubstituted amino acids incorporated into a single polypeptide. In some embodiments, the hydrocarbon crosslinkers are produced by a ring-closing metathesis reaction connecting two α,α-disubstituted amino acids. Ring-closing metathesis (also known as ring-closing olefin metathesis) is known in the art (Kim et al., Nature Protocols 6:761-771 (2011)).

[0651] The length of the hydrocarbon crosslinkers described herein can vary depending on the length of the substituents on the α,α-disubstituted amino acid. For example, by using a suitable α,α-disubstituted amino acid, the hydrocarbon linker produced by the ring-closing metathesis reaction can have a length of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 carbons. In some embodiments, the hydrocarbon linker has a length of 8 to 12 carbons. In some embodiments, the hydrocarbon linker has a length of 8 or 11 carbons. In certain embodiments, the hydrocarbon linker is 8 carbons long. In certain embodiments, the hydrocarbon linker is 11 carbons long. In some embodiments, the hydrocarbon crosslinker has a length of 2 to 15 carbons. In some embodiments, the hydrocarbon crosslinker has a length of 5 to 11 carbons. In some embodiments, the hydrocarbon crosslinker has a length of 7 to 11 carbons. In some embodiments, the hydrocarbon crosslinker has a length of 7 to 15 carbons. In some embodiments, the hydrocarbon crosslinker has a length of 8 to 11 carbons. In some embodiments, the hydrocarbon crosslinker has a length of 7, 8, 9, 10, 11 or more carbon atoms. In some embodiments, the hydrocarbon linker contains at least one double bond. In some embodiments, the hydrocarbon linker is an alkenyl crosslinker.

[0652] In some embodiments, stabilized polypeptides can be formed from any of the non-stapled polypeptides containing α,α-disubstituted amino acids described herein. In some embodiments, the hydrocarbon linker in the stabilized polypeptide is generated by reacting the α-alkenyl groups of at least two α-alkyl,α-alkenyl amino acids within the polypeptide structure. That is, the hydrocarbon linker is formed by reacting the α-alkenyl group of one amino acid with the α-alkenyl group of another amino acid to form an alkenyl hydrocarbon linker. In some embodiments, the reaction between the two α-alkenyl groups is a metathesis reaction. In some embodiments, the α-alkenyl group is a 4-pentenyl group or a 7-octenyl group. In some embodiments, in the stabilized polypeptide, one α-substituent in the α,α-disubstituted amino acid is methyl, and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker. In such stabilized polypeptides, there are at least two amino acids within the polypeptide backbone, each having a hydrocarbon linker as its α-substituent.

[0653] In some embodiments, the hydrocarbon linker has the formula: [ka] In the formula, each [ka] indicates the point of attachment of the hydrocarbon linker to the α-carbon atom of the α,α-disubstituted amino acid. In some aspects of these embodiments, the α-carbon atom is also substituted with a methyl group (e.g., the α,α-disubstituted amino acid is an α-substituted derivative of alanine). In some embodiments, the α-carbon of each α,α-disubstituted amino acid connected by the hydrocarbon linker has the S-configuration. In some embodiments, the α-carbon of each α,α-disubstituted amino acid connected by the hydrocarbon linker has the R-configuration. In some embodiments, when two α,α-disubstituted amino acids in a polypeptide are joined by a hydrocarbon linker, the α-carbon of one amino acid has the S-configuration and the α-carbon of the other amino acid has the R-configuration.

[0654] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0655] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0656] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0657] In some embodiments, the staple polypeptide comprises an amino acid sequence selected from the following: [Table 36] wherein Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid, one α-substituent in the α,α-disubstituted amino acid is methyl, and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the alpha carbon atom of Xaa6. (For example, Xaa3 and Xaa6 are each an alpha-substituted derivative of alanine.) In some embodiments, the alpha carbon of Xaa3 and the alpha carbon of Xaa6 are in the S-configuration.

[0658] In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0659] In some embodiments, the staple polypeptide comprises an amino acid sequence selected from the following: RXaa 16 L(N-methylQ)Xaa 10 LRXaa3IQRXaa6(CBA)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 88), RXaa 16 (Cpa)(N-methylQ)Xaa 10(Cpa)RXaa3IQRXaa6(Cpa)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 89), HRQRXaa 16 LQXaa 10 LRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 90), HRQRXaa 16 LQXaa 10 (Cpa)RXaa3IQRXaa6(Cpa)(2-Nal) (SEQ ID NO: 91), HRQRXaa 16 LQXaa 10 (Cha)RXaa3IQRXaa6(Cha)(2-Nal) (SEQ ID NO: 92), and LEHRERXaa 16 LQXaa 10 LRXaa3IQRXaa6L (sequence number 93). In the formula, Xaa 16 , Xaa 10 , Xaa3, and Xaa6 are each independently an α,α-disubstituted amino acid, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker having the formula: [ka] In the formula, each [ka] represents the hydrocarbon linker, Xaa 16 , Xaa 10 , Xaa3, or Xaa6 (one hydrocarbon linker is attached to the α carbon atom of Xaa 16 ~Xaa 10 and another hydrocarbon linker is between Xaa3 and Xaa6). In some embodiments, each hydrocarbon crosslinker has the formula: [ka]

[0660] In some embodiments, the staple polypeptide comprises an amino acid sequence selected from the following: HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 94), HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 95), HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-ClPh)(2-Nal) (SEQ ID NO: 96), and HRXaa 14 RSLXaa9TLRXaa3IQRXaa6(4-Cl-Ph)(2-Nal) (SEQ ID NO: 198), wherein the N-terminus of SEQ ID NO: 95 and SEQ ID NO: 96 is modified with palmitoyl-PEG4; In the formula, Xaa 14 , Xaa9, Xaa3, and Xaa6 are each independently an α,α-disubstituted amino acid, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker having the formula: [ka] In the formula, each [ka] represents the hydrocarbon linker, Xaa 14 , Xaa9, Xaa3, or Xaa6 (one hydrocarbon linker is attached to the α carbon atom of Xaa 14 to Xaa9, and another hydrocarbon linker is between Xaa3 and Xaa6). In some embodiments, each hydrocarbon crosslinker has the formula: [ka]

[0661] In some embodiments, the staple polypeptide comprises the following amino acid sequence: LQXaa 10 LRDIQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 97) In the formula, Xaa 10 and Xaa6 are each independently an α,α-disubstituted amino acid, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker having the formula: [ka] In the formula, each [ka] represents the hydrocarbon linker, Xaa 10 or indicates the point of attachment to the α carbon atom of Xaa6. In some embodiments, the hydrocarbon crosslinker has the formula: [ka]

[0662] In some embodiments, the staple polypeptide comprises an amino acid sequence selected from the following: [Table 37] wherein Xaa1, Xaa2, Xaa4, Xaa5, and Xaa7 are as described herein; and Xaa3 and Xaa6 are each independently an α,α-disubstituted amino acid, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the alpha carbon atom of Xaa6. (E.g., Xaa3 and Xaa6 are each an alpha-substituted derivative of alanine.) In some embodiments, the alpha carbon of Xaa3 and the alpha carbon of Xaa6 are in the S-configuration. In some embodiments, a stapled polypeptide comprising SEQ ID NO:1, SEQ ID NO:15, SEQ ID NO:30, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, or SEQ ID NO:31 may further comprise any additional amino acid or amino acid sequence, or combination thereof, as described herein for a non-stapled polypeptide comprising the same SEQ ID NO. For example, a stapled peptide may comprise any of the sequences disclosed in Table 3, or a combination thereof.

[0663] In some embodiments, the staple polypeptide consists of the following amino acid sequence: RSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 4), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0664] In some embodiments, the staple polypeptide consists of the following amino acid sequence: RERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 5), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0665] In some embodiments, the staple polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 6), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0666] In some embodiments, the staple polypeptide consists of the following amino acid sequence: EHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 7), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0667] In some embodiments, the staple polypeptide consists of the following amino acid sequence: QLEHRERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 8), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0668] In some embodiments, the staple polypeptide consists of the following amino acid sequence: EHRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 9), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0669] In some embodiments, the staple polypeptide consists of the following amino acid sequence: QERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 10), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0670] In some embodiments, the staple polypeptide consists of the following amino acid sequence: HQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 11), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0671] In some embodiments, the staple polypeptide consists of the following amino acid sequence: EHQERSLQTLRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 12), the staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0672] In some embodiments, the staple polypeptide consists of the following amino acid sequence: RERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 13), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0673] In some embodiments, the staple polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQRXaa6(CBA)(2-Nal) (SEQ ID NO: 14), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0674] In some embodiments, the staple polypeptide consists of the following amino acid sequence: AQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 21), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0675] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTARXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 22), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0676] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3AQRXaa6A(2-Nal) (SEQ ID NO: 23), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0677] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 24), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0678] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 25), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0679] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 26), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0680] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQAXaa6L(2-Nal)AA (SEQ ID NO: 27), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0681] In some embodiments, the staple polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQAXaa6L(2-Nal) (SEQ ID NO: 28), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0682] In some embodiments, the staple polypeptide consists of the following amino acid sequence: HRERSLQTLRXaa3IQAXaa6(CBA)(2-Nal) (SEQ ID NO: 29), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0683] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTARXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 75), LQTLRXaa3AQRXaa6L(2-Nal) (SEQ ID NO: 76), or LQTLRXaa3IQRXaa6A(2-Nal) (SEQ ID NO: 77), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0684] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)PD (SEQ ID NO: 32), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0685] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)P (SEQ ID NO: 33), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0686] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6, one α substituent of Xaa3 and Xaa6 is methyl (i.e., Xaa3 and Xaa6 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa6 are in the S-configuration.

[0687] In some embodiments, the staple polypeptide consists of the following amino acid sequence: LQTLRXaa3IQRXaa6L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34a), wherein the C-terminus of SEQ ID NO: 34a is modified with GRKKRRQRRRPQK(PEG4-palmitoyl)NH2; The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0688] In some embodiments, the staple polypeptide consists of the following amino acid sequence: QLEHRERSLXaa1TLRXaa3IQRML(2-Nal) (SEQ ID NO: 36), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, one α substituent of Xaa1 and Xaa3 is methyl (i.e., Xaa1 and Xaa3 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa1 and the α carbon of Xaa3 are in the S-configuration.

[0689] In some embodiments, the staple polypeptide consists of the following amino acid sequence: QLEHRERSLXaa1TLRXaa3IQR(2-Abu)L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 78), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3.

[0690] In some embodiments, the staple polypeptide consists of the following amino acid sequence: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal) (SEQ ID NO: 39), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa2, wherein one α substituent of Xaa1 and Xaa2 is methyl (i.e., Xaa1 and Xaa2 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa1 and the α carbon of Xaa2 are hydrocarbon linkers in the S-configuration, and a hydrocarbon linker having the formula: formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa4, one α substituent of Xaa3 and Xaa4 is methyl (i.e., Xaa3 and Xaa3 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa4 comprise a hydrocarbon linker in the S-configuration.

[0691] In some embodiments, the staple polypeptide consists of the following amino acid sequence: REXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 40), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa2, wherein one α substituent of Xaa1 and Xaa2 is methyl (i.e., Xaa1 and Xaa2 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa1 and the α carbon of Xaa2 are hydrocarbon linkers in the S-configuration, and a hydrocarbon linker having the formula: formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa4, one α substituent of Xaa3 and Xaa4 is methyl (i.e., Xaa3 and Xaa3 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa4 comprise a hydrocarbon linker in the S-configuration.

[0692] In some embodiments, the staple polypeptide consists of an amino acid sequence selected from the following: EXaa1SLQXaa2LRXaa3IQRXaa4L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 79), HREXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 82), HREXaa1SLQXaa2LRXaa3IQQXaa4(CBA)(2-Nal) (SEQ ID NO: 83), HR(N-methyl E)Xaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 84), HREXaa1SLQXaa2L(N-methylR)Xaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 85), HR(N-methylQ)Xaa1S(Cha)(N-methylQ)Xaa2(Cha)RXaa3IQRXaa4(Cha)(2-Nal) (SEQ ID NO: 86), and HRQXaa1SLQXaa2LRXaa3IQRXaa4(CBA)(2-Nal) (SEQ ID NO: 87), In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa1, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa2, one α substituent of Xaa1 and Xaa2 is methyl (i.e., Xaa1 and Xaa2 are each alanine substituted at the α position with a hydrocarbon linker), the α carbon of Xaa1 and the α carbon of Xaa2 are in the S-configuration, and the hydrocarbon linker has the formula: formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa4, one α substituent of Xaa3 and Xaa4 is methyl (i.e., Xaa3 and Xaa3 are each alanine substituted at the α position with a hydrocarbon linker), and the α carbon of Xaa3 and the α carbon of Xaa4 are in the S-configuration.

[0693] In some embodiments, the staple polypeptide consists of an amino acid sequence selected from the following: HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)L(2-Nal) (SEQ ID NO: 80), and HREXaa 15 SLQXaa 10 LR(Nle)IQR(Nle)(CBA)(2-Nal) (SEQ ID NO: 81), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] is Xaa of the hydrocarbon linker 15 indicates the point of attachment to the α carbon atom of [ka] is Xaa of the hydrocarbon linker 10 indicates the point of attachment to the α carbon atom of

[0694] In some embodiments, the staple polypeptide consists of an amino acid sequence selected from the following: LQTLEXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 98), LQTLRXaa3IQEXaa6L(2-Nal) (SEQ ID NO: 99), LQTLKXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 100), and LQTLRXaa3IQKXaa6L(2-Nal) (SEQ ID NO: 101), The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0695] In some embodiments, the staple polypeptide consists of an amino acid sequence selected from the following: (DL)QTIRXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 102), LQT(DL)RXaa3IQRXaa6L(2-Nal) (SEQ ID NO: 103), LQTLRXaa3(DI)QRXaa6L(2-Nal) (SEQ ID NO: 104), and LQTLRXaa3IQRXaa6(DL)(2-Nal) (SEQ ID NO: 105). The staple polypeptide comprises a hydrocarbon linker having the formula: [ka] In the formula, one [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa3, and the other [ka] indicates the point of attachment of the hydrocarbon linker to the α carbon atom of Xaa6.

[0696] In some embodiments, any one of the polypeptides described herein may have an N- or C-terminal chemical modification. For example, the N-terminus of the polypeptide may be modified with a moiety selected from acetyl, propionyl, hexanoyl, 3-phenylpropanoyl, 2-cyclohexylacetyl, diphenylacetyl, 3,5-dihydroxybenzoic acid, 4-(trifluoromethyl)benzoic acid, 5-phenylvaleric acid, 4-biphenylacetic acid, dimethyl, HOCH2CH2CO-, and palmitoyl-PEG4.

[0697] In some embodiments, the N-terminus is modified with Ac.

[0698] In some embodiments, the N-terminus is modified with palmitoyl-PEG4.

[0699] In some embodiments, the C-terminus of the polypeptide may be modified with a moiety selected from NH, (β-Ala)(β-Ala), (β-Ala)(β-Ala)NH, GRKKRRQRRRPQK(PEG4-palmitoyl)NH, GRKKRRQRRRPQNH, and 1-(2-aminoethyl)-4-methylpiperazine.

[0700] In some embodiments, the C-terminus of the polypeptide is modified at NH2.

[0701] In some embodiments, the C-terminus of the polypeptide is modified with (β-Ala)(β-Ala).

[0702] In some embodiments, the N-terminus of the polypeptide is modified with acetyl and the C-terminus of the polypeptide is modified with NH2.

[0703] In some embodiments, a salt of any of the polypeptides disclosed herein is formed between an acid and a base of the polypeptide, e.g., an amino functional group, or a base and an acidic group of the polypeptide, e.g., a carboxyl functional group. According to another embodiment, the polypeptide is a pharmaceutically acceptable acid addition salt.

[0704] In some embodiments, acids commonly used to form pharmaceutically acceptable salts of any of the polypeptides disclosed herein include inorganic acids such as hydrogen bisulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid, and organic acids such as paratoluenesulfonic acid, salicylic acid, tartaric acid, bitaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, parabromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, and acetic acid, and related inorganic and organic acids. Such pharmaceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, capenoate, heptanoate, propionate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, and the like. benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylate, citrate, β-hydroxybutynate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate, and other salts. In one embodiment, pharmaceutically acceptable acid addition salts include those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and especially those formed with organic acids such as maleic acid.

[0705] In some embodiments, bases commonly used to form pharmaceutically acceptable salts of any of the polypeptides disclosed herein include hydroxides of alkali metals, including sodium, potassium, and lithium; hydroxides of alkaline earth metals, such as calcium and magnesium; hydroxides of other metals, such as aluminum and zinc; ammonia, unsubstituted or hydroxyl-substituted mono-, di-, or tri-alkylamines, dicyclohexylamine; tributylamine; pyridine; N-methyl, N-ethylamine; N-ethylamine; diethylamine; triethylamine; mono-, bis-, or tris(2-OH-(C1-C6)-alkylamines), such as N-dimethyl-N-(2-hydroxyethyl)amine or tri-(2-hydroxyethyl)amine; n-methyl-d-glucamine; morpholine; thiomorpholine; organic amines, such as piperidine; and amino acids, such as arginine.

[0706] In some embodiments, the salt of the polypeptide is a trifluoroacetate salt, an acetate salt, or a hydrochloride salt. In some embodiments, the polypeptide or its pharmaceutically acceptable salt is substantially isolated. In some embodiments, the polypeptide or its pharmaceutically acceptable salt described herein is stable at 2-8°C for at least 1 month.

[0707] Peptide preparation and purification The present disclosure also provides methods for producing the polypeptides described herein. In some embodiments, methods for producing the polypeptides described herein include performing one or more chemical synthesis methods known to those skilled in the art and described herein. See, for example, Chapter 3 of Firds et al., Synthetic Peptides: A User's Guide, ed. Grant, W.H. Freeman & Co., New York, NY, 1992, p. 77; and Bird, G.H., et al., Methods Enzymol 446, 369-86 (2008). In some embodiments, methods for producing the polypeptides described herein include generating the polypeptide using solid-phase synthesis. For example, the polypeptides described herein can be produced by automated Merrifield techniques of solid-phase synthesis using side-chain protected amino acids and alpha-NH2 protected with either t-Boc or Fmoc chemistry, for example, on an Applied Biosystems peptide synthesizer model 430A or 431 or an AAPPTEC multichannel synthesizer APEX396.

[0708] In some embodiments, the polypeptides described herein can be produced in a high-throughput combinatorial manner, for example, using a high-throughput multi-channel combinatorial synthesizer. Other methods of synthesizing peptides are known in the art.

[0709] The method for producing a polypeptide described herein may further include forming one or more hydrocarbon linkers. One or more hydrocarbon linkers may be formed by subjecting a polypeptide containing at least two α,α-disubstituted amino acids described herein (e.g., Xaa1, Xaa2, Xaa3, Xaa4, or Xaa5) to metal-mediated ring-closing olefin metathesis. For example, synthetic strategies for generating hydrocarbon linkers based on modified Ala residues (α-methyl, α-alkenyl amino acids) will be known to those skilled in the art. The hydrocarbon linker connects adjacent turns of an α-helix with an α-methyl group at each end. The chemistry for generating this hydrocarbon linker is based on the incorporation of two α-methyl, α-alkenyl amino acids during peptide synthesis (see Kim 2011). Ruthenium-mediated ring-closing olefin metathesis is then used to generate a hydrocarbon linker between these modified amino acids. After ring closure, the polypeptide may be deprotected and released from such reactions, resulting in a polypeptide containing one or more hydrocarbon linkers.

[0710] The present disclosure also encompasses methods for purifying polypeptides produced according to the methods described herein. In some embodiments, the polypeptides are purified by high performance liquid chromatography (HPLC). In some embodiments, the purified polypeptides are substantially free of metals. As used herein, the term "substantially free of metals" refers to a composition comprising a polypeptide described herein and a metal at a concentration of less than about 0.5, 1, 2.5, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 ppm. In some embodiments, the purified polypeptide is substantially free of metals and contains less than about 0.5 ppm of metal. In some embodiments, the purified polypeptide contains less than about 5 ppm of metal. In some embodiments, the purified polypeptide contains less than about 20 ppm of metal.

[0711] How to use In some embodiments, administering a polypeptide (e.g., a stapled polypeptide) to a subject inhibits Wnt signaling in the subject. In some embodiments, administering a stabilized BCL9 peptide inhibits BCL9 binding to β-catenin. In some embodiments, administering a stabilized BCL9 peptide inhibits canonical Wnt / β-catenin signaling. In some embodiments, administering a stabilized BCL9 peptide treats a disease in a subject.

[0712] In some embodiments, the polypeptides described herein (e.g., stapled polypeptides) are capable of inhibiting BCL9 binding to β-catenin in vitro and / or in vivo. In some embodiments, polypeptides derived from the HD2 domain of a human BCL9 protein have one or more improved biological functions when compared to the non-stapled wild-type HD2 domain of a human BCL9 protein or when compared to a fragment of the non-stapled wild-type HD2 domain. The one or more biological functions can be selected from one or more of the following: (1) inhibiting BCL9 binding to β-catenin, (2) inhibiting canonical Wnt / β-catenin signaling, (3) decreasing regulatory T cell survival, (4) decreasing VEGF expression in tumors, (5) increasing CD4+ T cell and CD8+ T cell infiltration into tumors, (6) increasing T helper 17 (Th17) cells in tumors, (7) decreasing dendritic cells in tumors, (8) having a half-life (T1 / 2) of at least greater than two hours when administered to a subject, (9) inducing a tumor microenvironment favorable for an immune response, and (10) inhibiting tumor growth, cancer stem cell proliferation, and / or tumor metastasis.

[0713] In some embodiments, the polypeptides described herein exhibit favorable biological functions in some or each of the categories listed above, e.g., efficacy in various biochemical and cellular bioassays, including cell-based Wnt and / or β-catenin transcription assays.

[0714] For example, without being bound by any theory, the polypeptides described herein may have improved biological function in inhibiting BCL9 binding to β-catenin when assessed in various in vitro assays, e.g., in an alpha assay or a Wnt reporter assay, compared to a control polypeptide, e.g., the HD2 domain of non-stapled wild-type human BCL9. In this context, the polypeptides described herein may have improved K D or the polypeptides described herein may bind to β-catenin in the presence of a control polypeptide, indicating that the polypeptides described herein have an improved ability to inhibit BCL9 binding to β-catenin as compared to the control polypeptide. In some embodiments, the assays used to assess the biological function of the polypeptides described herein provide a quantitative measurement value(s), and the measurement observed with the disclosed polypeptides is at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, or more changed / improved as compared to that observed with a vehicle control polypeptide (e.g., the HD2 domain of unstapled wild-type human BCL9).

[0715] BCL9 binding to β-catenin In some embodiments, the polypeptides or variants described herein inhibit the binding of BCL9 to β-catenin in vitro and / or in vivo. In some embodiments, the polypeptides or variants disclosed herein inhibit the interaction of Pygo with BCL9 or the formation of a Pygo / BCL9 / β-catenin complex. Pygopus (Pygo) and Legless (Lgs) were discovered in Drosophila as novel Wnt signaling components essential for Armadillo-mediated transcription during normal development (see, e.g., Belenkaya et al., Development (2002) 129(17):4089-4101). Pygo and BCL9 / Legless transmit Wnt signals by promoting the transcriptional activity of β-catenin / Armadillo in normal and malignant cells. The ability of a polypeptide to inhibit the binding of BCL9 to β-catenin can be assessed by various assays known in the art. In some embodiments, the polypeptides described herein inhibit BCL9 binding to β-catenin as assessed by a homogeneous time-resolved fluorescence (HTRF) binding assay. In this assay, the polypeptide is conjugated to a tag capable of recognizing another tag bound to its target protein (i.e., β-catenin). When the polypeptide binds to the target protein, thereby bringing the two tags into close proximity, a signal is generated, which can be quantitatively read and used to calculate the binding affinity of the polypeptide. In some embodiments, the binding affinity of the polypeptide in this assay is compared to that of a control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9), and improved binding affinity compared to that of the control polypeptide is detected, indicating that the polypeptide is likely to inhibit BCL9 binding to β-catenin more efficiently than the control polypeptide. This assay may be performed in the presence or absence of an untagged control polypeptide.The assay may be performed by tagging a control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9) in the presence or absence of an untagged polypeptide described herein.

[0716] In some embodiments, the polypeptides described herein inhibit BCL9 binding to β-catenin as assessed by an amplified luminescence proximity homogeneous assay (ALPHA). In this assay, the polypeptide is conjugated to donor beads, and its target protein (i.e., β-catenin) is bound to acceptor beads. When the polypeptide binds to the target protein, bringing the two beads into close proximity, a signal is generated, allowing the binding affinity of the polypeptide to be quantitatively calculated. In some embodiments, the binding affinity of the polypeptide in this assay is compared to that of a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9), and improved binding affinity compared to the vehicle or control polypeptide is detected, indicating that the polypeptide is likely to inhibit BCL9 binding to β-catenin more efficiently than the control polypeptide. This assay may be performed in the presence or absence of an unconjugated control polypeptide. The assay may be performed by conjugating a control polypeptide in the presence or absence of an unconjugated polypeptide described herein.

[0717] In various embodiments, the polypeptides described herein inhibit BCL9 binding to β-catenin as assessed by a Wnt transcription assay. In some embodiments, the Wnt transcription assay is a cell-based assay. In some embodiments, the cell-based Wnt transcription assay is the GeneBLAzer® beta-lactamase (bla) reporter assay. Various cell lines, transformed cell lines, or primary cells derived from healthy or diseased subjects can be used in this assay. Cell lines known to be dependent on canonical Wnt / β-catenin signaling for survival may also be used. In some embodiments, CellSensor™ LEF / TCF-bla HCT-116 cells are used in this reporter assay. These cells contain a beta-lactamase (BLA) reporter gene under the control of a β-catenin / LEF / TCF response element stably integrated into the HCT-116 cells. Because these cells constitutively express beta-lactamase, adding a polypeptide that inhibits BCL9 binding to β-catenin in this assay leads to reduced beta-lactamase production. Thus, the efficiency of a polypeptide to inhibit Wnt transcription can be quantitatively calculated in this assay. In some embodiments, the polypeptides described herein inhibit Wnt transcription as measured in a GeneBLAzer® beta-lactamase (bla) reporter assay, which indicates the ability of the polypeptide to inhibit BCL9 binding to β-catenin. In some embodiments, the polypeptides tested in this assay exhibit an improved IC in inhibiting Wnt transcription compared to that of a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). 50 , indicating that the disclosed polypeptides are likely to inhibit BCL9 binding to β-catenin more efficiently than vehicle or control polypeptides.

[0718] In some embodiments, the polypeptides described herein inhibit BCL9 binding to β-catenin as assessed by a cell viability assay. In some embodiments, the cell viability assay is a CellTiterGlo luminescent assay, in which cell viability is quantitatively measured. Various cell lines, transformed cell lines, or primary cells derived from healthy or diseased subjects can be used in this assay. In some embodiments, the polypeptides described herein inhibit cell proliferation more efficiently in this assay than a vehicle or control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9), indicating that the disclosed polypeptides likely inhibit BCL9 binding to β-catenin more efficiently than a vehicle or control polypeptide.

[0719] Canonical Wnt / β-catenin signaling In certain embodiments, the polypeptides described herein can inhibit canonical Wnt / β-catenin signaling. Canonical Wnt / β-catenin signaling can be evaluated in in vitro and / or in vivo assays. In some embodiments, the effect of the polypeptides described herein on canonical Wnt / β-catenin signaling is evaluated in a cell-based Wnt transcription assay, such as the GeneBLAzer® beta-lactamase (bla) reporter assay. The GeneBLAzer® beta-lactamase (bla) reporter assay measures the strength of canonical Wnt / β-catenin signaling by its ability to regulate the β-catenin / LEF / TCF response element, and can therefore be used to evaluate whether a test agent can attenuate or increase the strength of canonical Wnt / β-catenin signaling regulation of its transcriptional target. In some embodiments, the polypeptides described herein suppress Wnt transcription as measured in a GeneBLAzer® beta-lactamase (bla) reporter assay, indicating that the polypeptides can inhibit canonical Wnt / β-catenin signaling. In some embodiments, the polypeptides in this assay exhibit an improved IC in suppressing Wnt transcription compared to that of a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). 50 , indicating that the polypeptides described herein have an improved ability to inhibit canonical Wnt / β-catenin signaling compared to vehicle or control polypeptides.

[0720] The ability of the polypeptides described herein to inhibit canonical Wnt / β-catenin signaling can also be assessed by measuring gene expression and / or protein expression of target genes transcriptionally regulated by canonical Wnt / β-catenin signaling. Target gene expression can be assessed in transformed cells contacted with a polypeptide described herein or in subjects administered such a polypeptide. Target genes include, for example, c-myc, ccnd1, cd44, LGR5, VEGFA, AXIN2, and LEF1. The expression levels of one or more target genes associated with canonical Wnt / β-catenin signaling can be analyzed using methods known in the art, such as cell staining, flow cytometry, Western blotting, and / or real-time quantitative PCR (rt-qPCR) analysis. In some embodiments, the polypeptides described herein reduce the expression of one or more target genes in cells. In some embodiments, the polypeptides described herein reduce the expression of one or more target genes more efficiently than a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0721] Regulatory T cell survival In some embodiments, the polypeptides described herein reduce the survival time of regulatory T cells. In some embodiments, when administered to a subject, the polypeptides reduce the survival time of regulatory T cells locally (e.g., in a tumor) and / or systemically (e.g., in the blood). In some embodiments, when administered to a subject, the polypeptides reduce the survival time of regulatory T cells compared to a control polypeptide. Various markers, such as CD4, FOXP3, and CD25, are known to be expressed on regulatory T cells. The ability of the polypeptides disclosed herein to reduce the survival time of regulatory T cells can be assessed by counting the total number of regulatory T cells present in a particular tissue, such as blood and / or tumor. For example, a sample obtained from a subject contacted with a polypeptide described herein may be stained with an antibody that detects a marker associated with regulatory T cells. The sample may be treated and labeled with an antibody that detects such a marker and analyzed by flow cytometry. The gene and / or protein expression of such markers can be determined in the sample and analyzed by Western blotting and / or rt-qPCR.

[0722] In some embodiments, the polypeptides described herein, when administered to a subject, reduce the number of regulatory T cells in the blood and / or tumor. In some embodiments, the polypeptides reduce the expression of one or more markers associated with regulatory T cells in one or more samples obtained from the subject to which the polypeptides are administered. In some embodiments, the polypeptides further reduce the expression of one or more markers compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9) when assessed in vivo.

[0723] VEGF expression in tumors In certain embodiments, the polypeptides described herein, when administered to a subject with a tumor, reduce VEGF expression in the tumor. Various assays can be used to measure VEGF gene expression and / or protein expression in tumor samples. For example, after contacting a subject with the polypeptide, tumor cells can be collected and stained with an anti-VEGF antibody to detect VEGF protein. Cells can also be analyzed, for example, by rt-qPCR, to determine VEGF gene expression. Other assays that indicate changes in VEGF expression can also be used. For example, tumor samples from subjects contacted with the polypeptides described herein can be analyzed to detect various angiogenesis markers regulated by VEGF. In some embodiments, the polypeptides described herein reduce VEGF expression more effectively than vehicle or a control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0724] Tumor infiltration of CD4+ and / or CD8+ T cells In some embodiments, the polypeptides described herein, when administered to a subject with a tumor, increase the infiltration of CD4+ T cells and / or CD8+ T cells into the tumor. Infiltration of CD4+ T cells and / or CD8+ T cells into the tumor can be assessed by counting the total number of CD4+ T cells and / or CD8+ T cells present in the tumor or a sample (e.g., a biopsy) from the tumor. In some embodiments, the polypeptides described herein, when administered to a subject with a tumor, increase the infiltration of CD4+ T cells and / or CD8+ T cells into the tumor more effectively than a vehicle or control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9). Various markers, such as CD4 and CD45, are known to be expressed on CD4+ T cells (also known as helper T cells). Various markers, such as CD8 and CD45, are known to be expressed on CD8+ T cells (also known as cytotoxic T cells). The ability of a polypeptide to increase the infiltration of CD4+ T cells and / or CD8+ T cells into a tumor can be evaluated in vivo by administering the polypeptide to a tumor-bearing subject. Tumor samples can be collected from the subject and stained with antibodies that detect markers associated with CD4+ / CD8+ T cells. For example, the sample can be treated and labeled with antibodies that detect such markers and analyzed, for example, by flow cytometry. Gene and / or protein expression of such markers can also be determined in the sample and analyzed by Western blotting and / or rt-qPCR. In some embodiments, the polypeptides described herein increase the total amount of CD4+ T cells and / or CD8+ T cells in the tumor compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0725] In some embodiments, the polypeptides described herein, when administered to a subject, increase the total number of CD4+ T cells and / or CD8+ T cells in the blood. A systemic increase in CD4+ T cells and / or CD8+ T cells may also indicate increased infiltration of CD4+ T cells and / or CD8+ T cells into a particular tissue, such as a tumor. In some embodiments, the polypeptides described herein increase the amount of circulating CD4+ T cells and / or CD8+ T cells in vivo compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0726] T helper 17 cell infiltration into tumors In some embodiments, the polypeptides described herein, when administered to a tumor-bearing subject, increase the infiltration of T helper 17 cells into the tumor. The infiltration of T helper 17 cells into the tumor can be assessed by counting the total number of T helper 17 cells present in the tumor. In some embodiments, the polypeptides described herein, when administered to a tumor-bearing subject, increase the infiltration of T helper 17 cells into the tumor compared to a vehicle or control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9). Various markers, such as IL-17, are known to be expressed on T helper 17 cells. The ability of a polypeptide to increase the infiltration of T helper 17 cells into a tumor can be assessed in vivo by administering the polypeptide to a tumor-bearing subject. A tumor sample can be collected from the subject and stained with, for example, an antibody that detects a marker associated with T helper 17 cells. The sample can be treated and labeled with an antibody that detects such a marker and analyzed by flow cytometry. The gene and / or protein expression of such markers can also be determined in the sample and analyzed by Western blotting and / or rt-qPCR. The sample may be analysed to detect the amount of IL-17 present in the sample.

[0727] In some embodiments, the polypeptides described herein, when administered to a subject, increase the total amount of T helper 17 cells in the blood. A systemic increase in T helper 17 cells may indicate increased infiltration of T helper 17 cells into a particular tissue, such as a tumor. A systemic increase in T helper 17 cells can be assessed by measuring the amount of IL-17 present in a blood sample collected from the subject. In some embodiments, the polypeptides increase the amount of circulating T helper 17 cells in a subject compared to a vehicle or a control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). In some embodiments, the polypeptides described herein increase the amount of circulating IL-17 in a subject compared to a control polypeptide.

[0728] Dendritic cells in tumors In some embodiments, the polypeptides described herein, when administered to a subject with a tumor, modulate dendritic cells present in the tumor. The number of dendritic cells present in the tumor can be assessed, for example, by staining the tumor with an antibody that recognizes one or more markers associated with dendritic cells. In some embodiments, the polypeptides described herein, when administered to a subject with a tumor, reduce the number of dendritic cells present in the tumor more effectively than a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). In some embodiments, the polypeptides described herein, when administered to a subject with a tumor, increase the number of dendritic cells present in the tumor more effectively than a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). Various markers, such as CD11c, are known to be expressed on dendritic cells. The ability of a polypeptide to reduce dendritic cells in a tumor can be assessed in vivo by administering the polypeptide to a subject. A tumor sample can be collected from the subject and stained with an antibody that detects a marker associated with dendritic cells. For example, the sample can be treated and labeled with an antibody that detects such a marker and analyzed, for example, by flow cytometry. The gene and / or protein expression of such markers is analyzed, for example, by Western blotting and rt-qPCR.

[0729] In some embodiments, the polypeptides described herein, when administered to a subject, reduce the total amount of dendritic cells in the blood. In some embodiments, the polypeptides described herein, when administered to a subject, increase the total amount of dendritic cells in the blood. A systemic reduction in dendritic cells may indicate that the amount of dendritic cells in a particular tissue, such as a tumor, is also reduced. In some embodiments, the polypeptides described herein reduce the amount of circulating dendritic cells in a subject compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). In some embodiments, the polypeptides described herein increase the amount of circulating dendritic cells in a subject compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0730] Biomarkers The present disclosure also encompasses the method of measuring at least one biomarker to monitor the therapeutic effectiveness of the polypeptide or pharmaceutical composition described herein, or to select subjects for treatment with such polypeptide or pharmaceutical composition.In some embodiments, the biomarker is one or more of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin.As used herein, active β-catenin refers to the non-phosphorylated form of β-catenin.

[0731] Various known methods can be used to measure the gene expression level and / or protein level of such biomarkers. For example, samples such as tumor biopsy, blood, plasma, serum, urine, amniotic fluid, synovial fluid, endothelial cells, leukocytes, monocytes, other cells, organs, tissues, bone marrow, lymph nodes, or spleen can be obtained from a subject treated with the present polypeptide or pharmaceutical composition. In some embodiments, the sample is a tumor biopsy in a subject. The sample obtained from a subject can be stained with one or more antibodies or other detection agents that detect such biomarkers. The sample can also, or alternatively, be processed to detect the presence of nucleic acids, such as mRNAs, encoding the biomarkers, for example, by rt-qPCR.

[0732] In some embodiments, a reduction in the gene expression and / or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin indicates the therapeutic efficacy of a polypeptide or pharmaceutical composition described herein. The expression levels of such biomarkers can be measured, for example, after administration of the polypeptide or pharmaceutical composition for 1 day, 2 days, 3 days, 4 days, 5 days, 1 week, or 2 weeks, or any period in between. In some embodiments, methods are disclosed that include measuring the level of one or more biomarkers after one or more administrations of a polypeptide or pharmaceutical composition described herein. In some embodiments, the method further includes continuing administration of the polypeptide or pharmaceutical composition if the biomarker level decreases. In some embodiments, the method further includes administering an increased dosage or increasing the frequency of subsequent administrations of a polypeptide or pharmaceutical composition described herein if the biomarker level does not decrease. In some embodiments, treatment is discontinued if the biomarker level does not decrease after the initial administration. In various embodiments, biomarker levels are also measured before the first administration of a polypeptide or pharmaceutical composition described herein and compared to levels after one or more administrations, and treatment efficacy and continued treatment steps are determined based on the change in biomarker level(s) from the pre-administration level(s).

[0733] In some embodiments, elevated gene expression and / or protein levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin indicate that the subject would benefit more from treatment with a polypeptide or pharmaceutical composition described herein than subjects without elevated gene expression and / or protein levels. In some embodiments, methods of treatment are disclosed that include selecting a patient with elevated biomarker levels and administering a polypeptide or pharmaceutical composition described herein.

[0734] In certain embodiments, subjects with elevated gene and / or protein expression levels of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin are selected for treatment with the polypeptides or pharmaceutical compositions described herein. In some embodiments, subjects with tumors are selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and / or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin. In some embodiments, subjects with tumors are selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and / or protein levels of BCL9. In some embodiments, subjects with tumors are selected for treatment after obtaining a tumor sample from the subject and identifying elevated gene and / or protein levels of CD44. In some embodiments, a subject suffering from a tumor is selected for treatment after obtaining a tumor sample from the subject and identifying elevated levels of active β-catenin gene and / or protein.

[0735] Half-life in the target In various embodiments, the polypeptides described herein have one or more improved pharmacokinetic parameters compared to a vehicle or control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9). Such pharmacokinetic parameters include, for example, maximum observed concentration (C max ), time to reach maximum concentration (T max ), terminal half-life (T 1 / 2 ), total body clearance (CL), volume of distribution (V z ), the area under the curve from the time of dosing to the last observable concentration (AUC 0-t ), area under the curve extrapolated from the time of administration to infinity (AUC 0-inf ), and bioavailability.

[0736] Methods for evaluating pharmacokinetics are known in the art.For example, blood samples from subjects administered with the polypeptides described herein can be obtained at 5 minutes, 1, 2, 4, 6, 8, 12, and 24 hours after administration.The concentration of the polypeptide in the blood samples can be analyzed by various analytical tools, for example, LC / MS.Based on the concentration of the polypeptide at each time point, pharmacokinetic parameters are calculated.As used herein, "maximum observed concentration (C max The term "maximum serum concentration" refers to the maximum serum concentration that a polypeptide achieves after administration. max Related to the concept of time to maximum concentration (T max The terminal half-life (T ) is the time it takes for a polypeptide to reach its maximum serum concentration. 1 / 2 ) and "Half-life (T 1 / 2 The terms "distribution volume" and "bioavailability" are used interchangeably and refer to the time it takes for a polypeptide to lose half of its serum concentration. Total body clearance (CL) refers to the volume of blood per unit time that is completely cleared of a polypeptide. The term "volume of distribution" refers to the theoretically calculated volume required to contain the total amount of a polypeptide administered to a subject at the same concentration as that observed in the blood. The term "bioavailability" refers to the extent and rate at which a drug is absorbed into a biological system or becomes available at the site of physiological activity. Bioavailability may be a function of some of the above-mentioned properties, including stability, solubility, immunogenicity, and pharmacokinetics, and can be assessed using methods known to those skilled in the art.

[0737] In some embodiments, the polypeptides described herein have an improved half-life in a subject compared to a control polypeptide. In some embodiments, the polypeptides have a half-life of at least 0.5, 1, 2, 3, 5, or more than 8 hours, or any period therebetween, when administered to a subject. In some embodiments, the polypeptides described herein have a half-life of at least 2 hours when administered to a subject. The pharmacokinetic parameters of the polypeptides can be evaluated in mammals, including, for example, mice, rats, or humans. The parameters can also be evaluated using various routes of administration, such as intravenous, intraperitoneal, subcutaneous, and intramuscular administration. In some embodiments, the pharmacokinetic parameters of the polypeptides described herein are evaluated in mice. In some embodiments, the pharmacokinetic parameters of the polypeptides described herein are evaluated in mice administered the polypeptide subcutaneously. In some embodiments, the pharmacokinetic parameters of the polypeptides described herein are evaluated in humans. In some embodiments, the pharmacokinetic parameters of the polypeptides described herein are evaluated in humans after subcutaneous administration.

[0738] A favorable tumor microenvironment for immune response In various embodiments, the polypeptides described herein induce a tumor microenvironment that is more favorable to an immune response than a vehicle or control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9).

[0739] Various parameters can be used to evaluate tumor microenvironment.For example, an increase in the ratio of cytotoxic T cells and regulatory T cells in and / or around tumor tissue can indicate that the tumor microenvironment is favorable for immune response.A decrease in the amount of dendritic cells and / or regulatory T cells in and / or around tumor tissue can also indicate that the tumor microenvironment is favorable for immune response.Other parameters include an increase in circulating T cells in peripheral blood, and an increase in the ratio of T helper 17 cells and regulatory T cells in and / or around tumor tissue.These parameters can indicate that the tumor microenvironment is favorable for immune response.

[0740] In some embodiments, the polypeptides described herein can increase the ratio of the amount of cytotoxic T cells to the amount of regulatory T cells in the tumor microenvironment. In some embodiments, the change in ratio caused by the polypeptide exceeds that caused by a vehicle or a control polypeptide (e.g., non-stapled wild-type HD2 domain human BCL9).

[0741] In some embodiments, the polypeptides described herein can increase the ratio of T helper 17 cells to regulatory T cells in the tumor microenvironment. In some embodiments, the change in ratio caused by the polypeptide exceeds that caused by a vehicle or control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9).

[0742] Tumor growth, cancer stem cell proliferation, and / or tumor metastasis Because Wnt signaling is a regulator of tumor growth, the efficacy of treatments that affect BCL9 binding to β-catenin, such as stabilized peptides of the HD2 domain of the BCL9 peptides described herein, can be evaluated in animal models.

[0743] The in vivo efficacy of stabilized BCL9 peptides can be evaluated in human cancer models, for example, using BALB / c nude mice, since xenografts of human cancer cells grow and develop into tumors in these mice. For example, subcutaneous inoculation of Colo320DM tumor cells, a commercially available cell line derived from human colon cancer tissue, can be used to form tumors in BALB / c nude mice. Additional in vivo models are also available for evaluating the in vivo efficacy of the polypeptides disclosed herein. For example, human DLD-1 colon cancer cells can be implanted into nude mice to evaluate tumor growth. The CT26 syngeneic mouse model of colon cancer can also be used because it allows for the evaluation of tumor growth in the context of an intact immune system. Other types of cancer cells, such as B16 melanoma, 4T1 breast cancer, Renca renal carcinoma, and Lewis lung cell carcinoma cells, can also be used in these known animal models to evaluate the in vivo efficacy of the polypeptides disclosed herein.

[0744] The polypeptides described herein can be administered to one or more animal models to assess the efficacy of the polypeptides in reducing tumor growth in vivo. In some embodiments, the polypeptides inhibit tumor growth in vivo more effectively than vehicle or a control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9). In some embodiments, the tumor burden / volume of subjects administered a polypeptide described herein is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% smaller than that of subjects administered a control polypeptide. From animal data on treatment with stabilized BCL9 peptides, the ability of the peptide to inhibit Wnt signaling can be assessed, for example, by staining tissue samples with markers of Wnt signaling. Such downstream markers of Wnt signaling include, for example, Axin2 and CD44.

[0745] An orthotopic mouse model may be used to evaluate the effect of the polypeptides described herein on tumor metastasis. For example, cells carrying a luciferase construct may be injected into the orthotopic animal model, followed by the assigned treatment. The presence of the injected cells may be detected by administering a luciferin substrate to each treated animal. The intensity of the bioluminescent signal may be quantitatively measured and used as an indicator of cell proliferation. In some embodiments, the polypeptides described herein suppress tumor metastasis more effectively than control polypeptides when evaluated in an orthotopic mouse model. In some embodiments, the polypeptides reduce tumor growth in an orthotopic mouse model by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% compared to vehicle or a control polypeptide (e.g., unstapled wild-type HD2 domain human BCL9).

[0746] In some embodiments, the effect of the polypeptides described herein on cancer stem cell proliferation can be assessed by measuring various cancer stem cell biomarkers. For example, the expression levels of CD44 and / or LGR5 can indicate the amount of cancer stem cells present in a sample. Tumor samples can be collected from a subject and stained with antibodies that detect markers associated with cancer stem cells. For example, the sample can be treated and labeled with antibodies that detect such markers and analyzed, for example, by flow cytometry. Gene and / or protein expression of such markers can be detected and analyzed, for example, by Western blotting and rt-qPCR. In some embodiments, the polypeptides described herein, when administered to a tumor-bearing subject, reduce the expression levels of CD44 and / or LGR5 in the tumor. In some embodiments, the polypeptides described herein reduce the expression levels of CD44 and / or LGR5 more effectively than a vehicle or control polypeptide (e.g., the unstapled wild-type HD2 domain of human BCL9 protein).

[0747] Diseases associated with abnormal Wnt / β-catenin signaling Aberrant Wnt / β-catenin signaling has been implicated in the malignant transformation of normal cells into cancerous cells (see Thakur 2013). Activation of Wnt signaling and nuclear localization of β-catenin have been linked to tumor phenotypes in multiple models.

[0748] The present disclosure encompasses compositions for use in inhibiting the binding of BCL9 to β-catenin in a subject by administering to the subject a staple polypeptide or a pharmaceutical composition comprising the polypeptide disclosed herein, and methods of using the same.The present disclosure also encompasses inhibiting canonical Wnt / β-catenin signaling in a subject by administering a polypeptide or pharmaceutical composition disclosed herein.The present disclosure further encompasses a method for treating a disease in a subject by administering a polypeptide or pharmaceutical composition described herein to a subject.The disease may be cancer or other neoplastic diseases associated with abnormal canonical Wnt / β-catenin signaling.

[0749] In some embodiments, the disease, disorder, or condition may be a disease that may benefit from the inhibition of canonical Wnt / β-catenin signaling. In some embodiments, such a disease, disorder, or condition is cancer. In some embodiments, the cancer is a cancer in which BCL9 and / or β-catenin are highly expressed. In some embodiments, the cancer is a cancer in which BCL9 and β-catenin are co-localized in the nucleus of cancer cells. In some embodiments, the cancer is selected from: familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, kidney cancer, brain / CNS cancer, throat cancer, multiple myeloma, cutaneous melanoma, acute lymphocytic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangioma, stomach cancer, ovarian cancer, hepatocellular carcinoma, and lymphangiogenesis. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is gastric cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is hepatocellular carcinoma. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is cutaneous melanoma. In some embodiments, the cancer is lung cancer.

[0750] In some embodiments, any of the polypeptides or variants disclosed herein, or pharmaceutical compositions comprising such polypeptides, may be used for the treatment of diseases, such as the cancers listed above.

[0751] In some embodiments, the tumor volume in a subject is reduced by more than 10%, 20%, 30%, 40%, or 50% (or any percentage therebetween) after administration of one or more doses of a polypeptide or pharmaceutical composition comprising a polypeptide described herein, compared to that of a subject treated with a vehicle or non-stapled peptide. In certain embodiments, this reduction is achieved after one week, two weeks, three weeks, or more (or any period therebetween). In some embodiments, the tumor volume in a subject is reduced by more than 50% after two weeks of administration, compared to that of a subject treated with a vehicle or non-stapled peptide. Since the materials and techniques required for various administration methods are available and known in the art, the appropriate dosage and / or formulation of a pharmaceutical composition for administration to a subject can be determined by one of skill in the art. For example, see Formulation and Delivery of Peptides and Proteins, 1 st edition, Washington, ACS, pp. 22-45 and Peptide and protein drug delivery, 1 st See, e.g., IEEE Transactions on Cancer, Vol. 1, No. 1, pp. 247-301, 2003. In some embodiments, the tumor volume of a subject administered a polypeptide or a pharmaceutical composition comprising the polypeptide is reduced by more than 50% after two weeks of administration compared to that of a subject treated with a vehicle or a wild-type polypeptide. In some embodiments, the tumor volume of a subject administered a polypeptide or a pharmaceutical composition comprising the polypeptide is reduced by 10% to more than 50% after two weeks of administration compared to that of a subject treated with a vehicle or a wild-type polypeptide.

[0752] Treatment and measurement parameters of treatment can be evaluated after the administration of the polypeptide or pharmaceutical composition alone or in combination with one or more additional therapeutic agents, for example, a single bolus or separate continuous administration.The additional agent can be any of the additional therapeutic agents mentioned herein or known to those skilled in the art.The polypeptide and / or pharmaceutical composition comprising the polypeptide, and / or the additional agent can be administered once or multiple times according to the selected regimen.

[0753] The present disclosure also encompasses a polypeptide or pharmaceutical composition disclosed herein for use in treating a subject's disease. In some embodiments, the disease may benefit from the inhibition of canonical Wnt / β-catenin signaling. In some embodiments, the disease is cancer.

[0754] The present disclosure further encompasses the use of the polypeptide or pharmaceutical composition disclosed herein in the manufacture of a medicament for treating a subject's disease.In some embodiments, the disease may benefit from the suppression of canonical Wnt / β-catenin signaling.In some embodiments, the disease is cancer.

[0755] In another embodiment, the disease to be treated is a disease other than cancer. In certain embodiments, the disease is abnormal bone density, ocular vascular disease, familial exudative vitreoretinopathy, premature coronary disease, Alzheimer's disease, autosomal dominant partial anodontia, retinal neovascularization, osteogenesis imperfecta, Tetra-Amelia syndrome, Müllerian duct regression and virilization, SERKAL syndrome, diabetes mellitus type II, Fuhrmann syndrome, odonto-onycho-dermal dysplasia, obesity, cleft hand / foot dysplasia, caudal duplication, odontogenesis, skeletal dysplasia, partial hypoplasia of the skin, autosomal recessive anonychia, neural tube defects, or sclerosteosis and Van Buchem disease.

[0756] Pharmaceutical Compositions, Formulations, Dosages, and Routes of Administration In various embodiments, pharmaceutical compositions are provided that comprise one or more of the polypeptides disclosed herein, alone or in combination with other prophylactic agents, therapeutic agents, and / or pharmaceutically acceptable carriers. In some embodiments, the pharmaceutical compositions may comprise one, two, three, or more of the polypeptides described herein. Pharmaceutical compositions comprising the polypeptides provided herein are intended for use in, but not limited to, the diagnosis, detection, or monitoring of disorders, the prevention, treatment, or amelioration of disorders or one or more symptoms thereof, and / or research.

[0757] "Pharmaceutically acceptable carrier" refers to any and all solvents, solid, semi-solid, liquid fillers, diluents, encapsulating materials, formulation aids, vehicles, isotonicity agents, and absorption delaying agents that are used in combination with the polypeptides described herein to form, for example, a "pharmaceutical composition" suitable for administration to a subject. The use of such media and agents for pharmaceutically active substances is well known in the art. Supplementary active compounds can also be incorporated into the composition. The pharmaceutically acceptable carrier can be selected based on the use and / or route of administration of the composition.

[0758] The pharmaceutical composition may be formulated into any of many possible dosage forms, such as, for example, tablets, capsules, gel capsules, powders, or granules. The pharmaceutical composition may be formulated as a solution, suspension, emulsion, or mixed medium. In some embodiments, the pharmaceutical composition may be formulated as a lyophilized preparation or aqueous solution suitable for administration, for example, by injection or infusion.

[0759] In some embodiments, the pharmaceutical composition may be formulated as a solution. For example, the polypeptides described herein may be administered in an unbuffered solution, such as saline, water, or dimethyl sulfoxide (DMSO). In some embodiments, the polypeptides may be administered in a suitable buffer solution. For example, the buffer solution may contain acetate, citrate, prolamin, carbonate, or any combination thereof. In some embodiments, the buffer solution may be phosphate-buffered saline (PBS). The pH and osmolality of the buffer solution containing the polypeptide may be adjusted to be suitable for administration to a subject.

[0760] In some embodiments, the pharmaceutical composition may be formulated as a suspension in an aqueous medium, a non-aqueous medium, or a mixed medium. In some embodiments, the pharmaceutical composition is formulated in a mixed medium containing water and DMSO. Aqueous suspensions may further contain substances that increase the viscosity of the suspension, such as carboxymethylcellulose, sorbitol, and / or dextran. Suspensions may also contain stabilizers.

[0761] In some embodiments, the pharmaceutical compositions are used for in vivo administration and may be sterile, which can be readily accomplished, for example, by filtration through sterile filtration membranes.

[0762] In various embodiments, the pharmaceutical compositions comprising the polypeptides described herein may further comprise at least one additional agent. In some embodiments, the at least one additional agent is selected from one or more of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, and an anti-cancer agent.

[0763] In some embodiments, the pharmaceutical compositions described herein comprise a checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody. In one embodiment, the checkpoint inhibitor targets a stimulatory checkpoint molecule, such as CD27, CD40, OX40, GITR, or CD138. In yet another embodiment, the checkpoint inhibitor targets an inhibitory checkpoint molecule, such as A2AR, B7-H3, B7-H4, attenuator of B and T lymphocytes (BTLA), indoleamine 2,3-dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG3), T cell immunoglobulin domain and mucin domain 3 (TIM-3), VISTA (C10orf54), or V-domain Ig suppressor of T cell activation.

[0764] In some embodiments, the pharmaceutical compositions described herein comprise an EGFR inhibitor. In one embodiment, the EGFR inhibitor is erlotinib, gefitinib, lapatinib, panitumumab, vandetanib, or cetuximab.

[0765] In some embodiments, the pharmaceutical compositions described herein comprise a VEGF or VEGFR inhibitor.In one embodiment, the VEGF or VEGFR inhibitor is pazopanib, bevacizumab, sorafenib, sunitinib, axitinib, ponatinib, regorafenib, vandetanib, cabozantinib, ramucirumab, lenvatinib or ziv-aflibercept.

[0766] In some embodiments, the pharmaceutical compositions described herein comprise an anti-cancer drug, such as cyclophosphamide, methotrexate, 5-fluorouracil (5-FU), doxorubicin, mustine, vincristine, procarbazine, prednisolone, dacarbazine, bleomycin, etoposide, cisplatin, epirubicin, capecitabine, folinic acid, actinomycin, all-trans retinoic acid, azacitidine, azathioprine, bortezomib, carboplatin, chlorambucil, or the like. The agent may be selected from fluticasone, cytarabine, daunorubicin, docetaxel, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, paclitaxel, pemetrexed, teniposide, thioguanine, topotecan, valrubicin, vinblastine, vindesine, vinorelbine, and oxaliplatin.

[0767] Polypeptides of the present disclosure may be administered to a patient by topical (including ocular and mucosal membranes, including vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including nebulizers, intratracheal, intranasal), epithelial, transdermal, oral, or parenteral routes. Parenteral administration includes intravenous, subcutaneous, intraperitoneal, or intramuscular injection or infusion, or intracranial, e.g., intrathecal or intraventricular, administration. In some embodiments, a polypeptide or pharmaceutical composition described herein is administered intravenously. In some embodiments, a polypeptide or pharmaceutical composition described herein is administered intraperitoneally. In some embodiments, a polypeptide or pharmaceutical composition described herein is administered daily, weekly, monthly, or at any suitable interval that can be used to treat a disease of interest.

[0768] In the pharmaceutical compositions of the present application, the polypeptide is present in an effective amount (e.g., a therapeutically effective amount). The effective amount may vary depending on the disease being treated, the severity of the disease, the route of administration, the sex, age, and general health of the subject, the use of excipients, the possibility of coadministration with other therapeutic therapies such as the use of other drugs, and the judgment of the treating physician. In some embodiments, an effective amount of the polypeptide may range, for example, from about 0.001 mg / kg to about 500 mg / kg. Dosage regimens may be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, or several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in unit dosage form for ease of administration and uniformity of dosage. An exemplary, non-limiting range for a therapeutically or prophylactically effective amount of the binding proteins provided herein is 0.1 to 20 mg / kg, e.g., 1 to 10 mg / kg.

[0769] Combination therapy In certain embodiments, the polypeptide or pharmaceutical composition disclosed herein is administered together with at least one additional agent. That is, the polypeptide of the present disclosure and the additional agent can be administered to a patient sequentially or simultaneously in separate dosage forms as described herein. In some embodiments, the at least one additional agent is selected from a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a VEGFR inhibitor, an anti-cancer agent (for example, any of the additional therapeutic agents described herein). The staple peptide and the additional agent can be administered in a therapeutically effective amount.

[0770] In certain embodiments, a subject administered a polypeptide or pharmaceutical composition disclosed herein is also treated with radiation therapy and / or chemotherapy before, after, or simultaneously with administration of the polypeptide or pharmaceutical composition.

[0771] kit The present invention also includes pharmaceutical kits comprising one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a polypeptide of the present disclosure, useful, for example, for treating the disorders, diseases, and conditions mentioned herein. Such kits can optionally further include one or more of a variety of conventional pharmaceutical kit components, such as, for example, a container with one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions, either as an insert or label, indicating the amounts of components to be administered, administration guidelines, and / or guidelines for mixing the components, can also be included in the kit.

[0772] Also disclosed herein are kits for carrying out the methods described herein. In various embodiments, kits are provided for producing the polypeptides described herein. In some embodiments, the kits include polypeptides capable of undergoing a reaction to form one or more hydrocarbon linkers. In some embodiments, the kits include a metal catalyst for performing metal-mediated ring-closing olefin metathesis.

[0773] In some embodiments, the kit comprises agents for detecting gene and / or protein expression of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active β-catenin.

[0774] definition When the terms "contain," "include," "have," or grammatical variations of such terms are used in either this disclosure or the claims, such terms are intended to be inclusive in the same manner as the interpretation of "comprising" when the term "comprising" is used as a transitional term in the claims.

[0775] The term "about" means an amount, level, value, number, frequency, proportion, dimension, size, amount, weight, or length that differs by 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1% from the reference amount, level, value, number, frequency, proportion, dimension, size, amount, weight, or length. When the term "about" is used in conjunction with a numerical range, the term modifies that range by extending both the boundaries above and below the stated numerical values. In general, the term "about" is intended to modify numerical values that differ by ≦10% above and below the stated value.

[0776] As used herein, the term "amino acid" generally refers to an organic compound containing an amine (-NH2) and a carboxyl (-COOH) functional group, along with a side chain (R group) specific to each amino acid. The side chain can be hydrophobic or hydrophilic, charged or neutral, and aliphatic or aromatic. In naturally occurring amino acids, the amine and carboxyl functional groups are attached to the same carbon atom, i.e., the amino group is attached to the carbon at the alpha position relative to the carboxyl group. Any of the amino acids described herein may be in the L-configuration or the D-configuration. In some embodiments, the amino acid is in the L-configuration. In some embodiments, the amino acid is in the D-configuration. The 20 naturally occurring amino acids are abbreviated herein as shown in Table A. [Table 38]

[0777] Unnatural amino acids are referred to herein as follows:

[0778] Norleucine (abbreviated herein as Nle) is (2S)-2-aminohexanoic acid, which has the following chemical structure: [ka] .

[0779] β-alanine (or beta-alanine, abbreviated herein as β-Ala) is a 3-aminopropanoic acid having the following chemical structure: [ka] .

[0780] 2-Naphthylalanine (abbreviated herein as 2-Nal) is (S)-2-amino-3-(naphthalen-1-yl)propanoic acid, which has the following chemical structure: [ka] .

[0781] D-2-naphthylalanine (abbreviated herein as 2-Dnal) is (R)-2-amino-3-(naphthalen-1-yl)propanoic acid, which has the following chemical structure: [ka] .

[0782] β-leucine (abbreviated herein as β-L) is (R)-3-amino-4-methylpentanoic acid, which has the following chemical structure: [ka] .

[0783] β-Cyclobutylalanine (abbreviated herein as CBA) is (S)-2-amino-3-cyclobutylpropanoic acid, having the following chemical structure: [ka] .

[0784] N-methyl-D-glutamine (abbreviated herein as N-methylQ or NMeGln) is methyl-L-glutamine having the following chemical structure: [ka] .

[0785] N-methylarginine (abbreviated herein as N-methylR, NMeArg, or R(Me)) is a compound of the following formula: [ka] .

[0786] Cyclohexyl L-alanine (abbreviated herein as Cha) is (S)-2-amino-3-cyclohexylpropanoic acid of the following formula: [ka] .

[0787] Cyclohexyl D-alanine (abbreviated herein as Dcha) is (R)-2-amino-3-cyclohexylpropanoic acid of the following formula: [ka] .

[0788] N-methylcyclohexylalanine (abbreviated herein as NMeCha) is (S)-2-amino-3-cyclohexylpropanoic acid of the following formula: [ka] .

[0789] N-methylleucine (abbreviated herein as MeL, NMeLeu, or N-methyl L) is the methyl-L-leucine compound of the following formula: [ka] .

[0790] N-methylhistidine (abbreviated herein as N-methylH or N-MeHis) is the methyl-L-histidine compound of the following formula: [ka] .

[0791] Cyclopropyl-L-alanine (abbreviated herein as Cpa) is the (S)-2-amino-3-cyclopropylpropanoic acid compound of the following formula: [ka] .

[0792] N-methyl l-glutamic acid (abbreviated herein as N-methyl E) is the methyl-L-glutamic acid compound of the following formula: [ka] .

[0793] N-methylaspartic acid (abbreviated herein as N-methyl D) is a compound of the formula: [ka] .

[0794] N-methyl l-threonine (abbreviated herein as N-methyl T) is a compound of the following formula: [ka] .

[0795] D-threonine (abbreviated herein as DThr) is a compound of the formula: [ka] .

[0796] N-methyl-L-isoleucine (abbreviated herein as N-methyl-I) is a compound of the formula: [ka] .

[0797] N-methyl-L-cysteine (abbreviated herein as N-MeCys) is a compound of the following formula: [ka] .

[0798] Guanidino-L-phenylalanine (abbreviated herein as Phe(4-guanidino)) is the (S)-2-amino-3-(4-((diaminomethylene)amino)phenyl)propanoic acid compound of the following formula: [ka] .

[0799] Homoarginine (abbreviated herein as homoR or homoArg) is the compound of the formula: [ka] .

[0800] Homohistidine (abbreviated herein as homoH or homoHis) is the compound of the formula: [ka] .

[0801] Citrulline (abbreviated herein as Cit) is an unnatural amino acid of the following formula: [ka] .

[0802] 2-Aminobutyric acid (abbreviated herein as 2Abu or 2-Abu) is the (S)-2-aminobutanoic acid compound of the following formula: [ka] .

[0803] Tertorucine (abbreviated herein as Tle) is the (S)-2-amino-3,3-dimethylbutanoic acid compound of the following formula: [ka] .

[0804] 4-Chlorophenylalanine (abbreviated herein as 4-ClPh or Phe(4-Cl)) is the (S)-2-amino-3-(4-chlorophenyl)propanoic acid compound of the following formula: [ka] .

[0805] 3,4-Chlorophenylalanine (abbreviated herein as 3,4-diClPh) is the (S)-2-amino-3-(3,4-dichlorophenyl)propanoic acid compound of the following formula: [ka] .

[0806] 4-Fluorophenylalanine (abbreviated herein as 4-FPh or Phe(4-F)) is the (S)-2-amino-3-(4-fluorophenyl)propanoic acid compound of the following formula: [ka] .

[0807] L-α-neopentylglycine (abbreviated herein as NptGly), or t-butylalanine (abbreviated herein as β-tBu-Ala or tBua), is a compound of the following formula: [ka] .

[0808] Alpha-methylleucine (abbreviated herein as alpha-methyl L) is the (S)-2-amino-2,4-dimethylpentanoic acid compound of the following formula: [ka] .

[0809] t-Butylglycine (abbreviated herein as t-Bug) is a compound of the following formula: [ka] .

[0810] Azetidine-3-carboxylic acid (abbreviated herein as Aze) is a compound of the formula: [ka] .

[0811] Allylglycine (abbreviated herein as Alg or allylGly) is a compound of the formula: [ka] .

[0812] 5,5,5-trifluoroleucine (abbreviated herein as Tfl) is a compound of the formula: [ka] .

[0813] 1-Aminocyclobutane-1-carboxylic acid (abbreviated herein as AC4C) is a compound of the formula: [ka] .

[0814] 1-Aminocyclohexane-1-carboxylic acid (abbreviated herein as A6C) is a compound of the formula: [ka] .

[0815] Norarginine (abbreviated herein as Nar) is the compound (S)-2-amino-4-((diaminomethylene)amino)butanoic acid of the following formula: [ka] .

[0816] L-β-homotryptophan (abbreviated herein as β-homoTrp) is a compound of the formula: [ka] .

[0817] Cyclohexylmethylalanine (abbreviated herein as HomoCha) is the compound (S)-2-amino-4-cyclohexylbutanoic acid of the following formula: [ka] .

[0818] Hydroxyproline (abbreviated herein as Hyp) is a compound of the formula: [ka] .

[0819] (5-amino-5-oxopentyl)glycine (abbreviated herein as peptoid Q) is a compound having the formula: [ka] .

[0820] His peptoid (abbreviated herein as NHis) is the compound of the following formula ((1H-imidazol-4-yl)methyl)glycine: [ka] .

[0821] As used in connection with the polypeptides described herein, a "variant" refers to a polypeptide that differs from a given polypeptide in amino acid sequence and / or chemical structure, but retains one or more biological functions of the given polypeptide (i.e., a polypeptide described herein). For example, a variant can retain one or more biological functions of a polypeptide derived from the HD2 domain of the human BCL9 protein, such as the ability to bind to β-catenin, inhibit canonical Wnt / β-catenin signaling, and / or inhibit BCL9 binding to β-catenin. The variant polypeptides described herein can have one or more amino acid additions (e.g., insertions), deletions, and / or substitutions compared to the given polypeptide, so long as they retain the functional properties mentioned above. In some embodiments, the variant polypeptides described herein may have between 1 and 30, 1 and 20, 1 and 10, 1 and 8, 1 and 5, 1 and 4, 1 and 3, or 1 and 2, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or 30 (including all integers between these ranges) amino acid additions (e.g., insertions), deletions, and / or substitutions) relative to the wild-type polypeptide.

[0822] The term "variant" includes polypeptides having a certain percentage of homology to a wild-type polypeptide or fragment, such as at least about 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% (or any percentage therebetween). As used herein, the term "percent homology" defines the percentage of residues in the amino acid sequences of a variant and a given polypeptide that are identical after aligning the sequences and other spatial arrangements, for example, using BLAST alignment software. In some embodiments, variants include polypeptides that are chemically and / or post-translationally modified differently from the wild-type polypeptide or fragment but retain one or more of the biological functions described above. For example, variants may contain one or more amino acids that are post-transitionally modified, for example, by phosphorylation, acetylation, methylation, ubiquitination, sumoylation, or other post-translational modifications known in the art. Variants may also contain one or more chemical modifications, such as one or more amino acid side chains that are modified or replaced with different chemical moieties.

[0823] As used herein, the terms "hydrocarbon crosslinker" and "crosslinker" (also known as hydrocarbon staple, hydrocarbon linker, or metathesized crosslinker) are used interchangeably and refer to a chemical linker between two amino acids that significantly enhances and / or strengthens the secondary structure of a given polypeptide. The hydrocarbon crosslinkers described herein may be based on the incorporation of natural or unnatural amino acids that restrict the structural flexibility of the polypeptide compared to the wild-type (i.e., uncrosslinked) peptide.

[0824] As used herein, the term "polypeptide" refers to a sequence of amino acids chemically joined by covalent peptide (amide) bonds. That is, the amino group of one amino acid reacts with the carboxyl group of another amino acid to form an amide bond (peptide bond) between the amino acids. Typically, a polypeptide consists of 5 to 50 amino acid monomers, including an N-terminal amino acid and a C-terminal amino acid. In some embodiments, the terminal amino acid residues are unmodified, i.e., the polypeptide contains an amino group at one terminus and a carboxyl group at the other terminus. In some embodiments, the N-terminus and / or C-terminus of the polypeptide or variant are further modified. In some embodiments, the N-terminus is modified with an acetyl group. In some embodiments, the C-terminus is modified with an NH group. In some embodiments, the N-terminus and / or C-terminus modification further comprises a fluorenylmethyloxycarbonyl (Fmoc) group.

[0825] As used herein, the terms "alkyl" and "alkyl-" used alone or in combination with other terms are intended to mean the same or similar groups. Cn-m The term "alkyl" refers to a saturated hydrocarbon group that can be straight-chain (straight-chain) or branched, having n to m carbons. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like. In some embodiments, an alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

[0826] As used herein, "alkylene" and "C n-mThe term "alkylene" refers to a divalent saturated branched or straight-chain (straight-chain) chemical group containing only carbon and hydrogen atoms, such as methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, sec-butylene, tert-butylene, n-pentylene, isopentylene, sec-pentylene, and neopentylene. An alkylene group can be unsubstituted or substituted with one or more substituents. In some embodiments, an alkylene group contains 1 to 9 carbon atoms (e.g., 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 2 carbon atoms).

[0827] The terms "pharmaceutical" and "pharmaceutically acceptable" are used herein to refer to compounds, substances, compositions, and / or dosage forms that are suitable for use in contact with the tissues of human beings and animals, within the scope of sound medical judgment, and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0828] As used herein, the term "cell" is meant to refer to a cell that is in vitro, ex vivo, or in vivo. In some embodiments, an ex vivo cell may be part of a tissue sample excised from an organism, such as a mammal. In some embodiments, an in vitro cell may be a cell in cell culture. In some embodiments, an in vivo cell is a cell that lives within an organism, such as a mammal.

[0829] As used herein, the terms "individual," "patient," or "subject" refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and most preferably humans.

[0830] As used herein, the phrase "effective amount" or "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent that elicits the biological or drug response in a tissue, system, animal, individual, or human that is desired by a researcher, veterinarian, physician, or other clinician.

[0831] As used herein, the term "treating" or "treatment" refers to 1) inhibiting a disease, e.g., inhibiting a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., preventing further progression of the pathology or symptoms), or 2) ameliorating a disease, e.g., ameliorating a disease, condition, or disorder in an individual experiencing or exhibiting the pathology or symptoms of the disease, condition, or disorder (i.e., reversing the pathology or symptoms).

[0832] As used herein, the term "preventing" or "prevention" of a disease, condition, or disorder refers to reducing the risk of developing a disease, condition, or disorder in a subject or group of subjects (e.g., a subject or group of subjects prone to or susceptible to a disease, condition, or disorder). In some embodiments, preventing a disease, condition, or disorder refers to reducing the likelihood of acquiring the disease, condition, or disorder and / or its associated symptoms. In some embodiments, preventing a disease, condition, or disorder refers to completely or nearly completely halting the development of a disease, condition, or disorder.

[0833] As used herein, the terms "improve," "increase," "enhance," "elevate," "upregulate," and "promote" one or more biological functions are all used interchangeably and mean that the level or activity of one or more biological functions, or the measurement of those functions in in vitro and / or in vivo assays, is increased above the level or activity observed in the absence of a polypeptide described herein and / or is higher than a vehicle or control polypeptide (e.g., the HD2 domain of unstapled wild-type human BCL9, a polypeptide that does not contain the core functional domain that mediates the interaction of BCL9 with β-catenin, or a control polypeptide that includes a sequence not derived from the HD2 domain of human BCL9).

[0834] As used herein, the term "tumor microenvironment" refers to the cellular microenvironment within and / or surrounding a tumor, including tumors, blood vessels, immune cells, signaling molecules, and various cells recruited to the extracellular matrix. See, for example, Balkwill et al., J Cell Sci (2012) 125:5591-5596. The polypeptides described herein can alter the composition of immune cells and / or signaling molecules within and / or surrounding a tumor, thereby inducing an immune response in the peritumoral microenvironment. [Example]

[0835] material and method Each polypeptide used in the following examples (including the stabilized polypeptide) was produced by a single on-resin synthesis method. Each polypeptide was produced by on-resin peptide elongation. In the case of stapled polypeptides, peptide backbone synthesis was followed by a ring-closing metathesis reaction to produce the hydrocarbon linker.

[0836] Methods for synthesizing hydrocarbon linkers using modified Ala residues (α,α-disubstituted amino acids such as α-methyl, α-alkenyl amino acids) are known in the art. See, e.g., US2014 / 0113857 and Kim 2011.

[0837] Hydrocarbon linkers of different lengths, such as 8-carbon and 11-carbon crosslinkers, can be generated using α-methyl, α-alkenyl amino acids with alkenyl chains of the appropriate length. For example, (S)2-(4'-pentenyl)Ala was incorporated into the polypeptide to construct stabilized polypeptides with 8-carbon crosslinkers having an S configuration at both termini. (R)2-(4'-pentenyl)Ala was incorporated into the polypeptide to construct stabilized polypeptides with 8-carbon crosslinkers having an R configuration at both termini. (S)2-(4'-pentenyl)Ala and (R)2-(4'-pentenyl)Ala were used for stabilized polypeptides with 8-carbon crosslinkers having an S configuration at one terminus and an R configuration at the other terminus, respectively. To construct stabilized polypeptides with 11-carbon crosslinkers with an S configuration at one end and an R configuration at the other, (R)2-(7'-octenyl)Ala and (S)2-(4'-pentenyl)Ala were used, respectively.

[0838] Each polypeptide was purified using a standard high-performance liquid chromatography (HPLC) protocol. A Zorbax C18 reverse-phase column, 9.4 × 250 mm (Agilent, 80 Å pore size, 3.5 μm particle size) was used. The solvents used were A: water, 0.1% (vol / vol) TFA; B: acetonitrile, 0.1% (vol / vol) TFA. The flow rate was 4 ml / min. The gradient was 10–100% (vol / vol) B over 30 min, 100% B over 5 min, 100–10% (vol / vol) B over 4 min, and 10% (vol / vol) B over 1 min. The injection volume was 100–400 μl. The wavelength (nm) was 280 (for Fmoc-, Trp-, or Tyr-containing peptides) or 220 (for other peptides).

[0839] Example 1 - Non-stapled polypeptides containing α-monosubstituted unnatural amino acids The following peptides were prepared according to methods and procedures similar to those described in the Materials and Methods section. [Table 39]

[0840] Example 2 - Non-stapled polypeptides containing α,α-disubstituted amino acids The following peptides were prepared using methods and procedures similar to those described in the Materials and Methods section, and (S)-2-(4'-pentenyl)alanine was used as the base for each peptide. [ka] and [ka] This is the position. [Table 40]

[0841] Example 3 - Stapled Polypeptides The following peptides were prepared according to methods and procedures similar to those described in the Materials and Methods section by metathesis reactions forming hydrocarbon linkers of the formula: [ka] [ka] Between each pair of [ka] is a hydrocarbon linker, [ka] to the α carbon atom of [ka] The α carbon of is in the S configuration. [Table 41] TIFF2025122018000297.tif233158TIFF2025122018000298.tif233161TIFF2025122018 000299.tif233159TIFF2025122018000300.tif234158TIFF2025122018000301.tif70158 1 Unless otherwise specified, the N-terminus of the peptides is modified with an Ac group and the C-terminus is modified with an NH2 group. 2 The C-terminus is modified with GRKKRRQRRRPQK(PEG4-palmitoyl)NH2 。 3 The N-terminus is modified with HOCH2CH2CO-. 4 The N-terminus is modified with propionyl. 5 The N-terminus is modified with hexanoyl. 6 The N-terminus is modified with 3-phenylpropanoyl. 7 The N-terminus is modified with 2-cyclohexylacetyl. 8 The N-terminus is modified with diphenylacetyl. 9 The N-terminus is modified with 3,5-dihydroxybenzoic acid. 10 The N-terminus is modified with 4-(trifluoromethyl)benzoic acid. 11 The N-terminus is modified with 5-phenylvaleric acid. 12 The N-terminus is modified with 4-biphenylacetic acid. 13 The N-terminus is modified with dimethyl. 14 The N-terminus and C-terminus are unmodified. 15 The C-terminus is unmodified. 16 The N-terminus is modified with palmitoyl-PEG4. 17 The C-terminus is modified with GRKKRRQRRRPQ-NH2. 18 The C-terminus is modified with 1-(2-aminoethyl)-4-methylpiperazine. 19 The C-terminus is modified with K(PEG4-palmitoyl)NH2.

[0842] Example 4 - Alpha Assay Inhibition of BCL9 binding to β-catenin was assessed using an amplified luminescence proximity homogeneous assay (ALPHA). In this assay, a polypeptide is conjugated to donor beads, and its target protein (i.e., β-catenin) is bound to acceptor beads. When the polypeptide binds to the target protein, bringing the two beads into close proximity, a signal is generated, allowing the binding affinity of the polypeptide to be quantitatively calculated. This assay can also be performed in the presence or absence of an unconjugated control polypeptide. The results of the ALPHA assay are shown in Table 4a. [Table 42] TIFF2025122018000303.tif208163TIFF2025122018000304.tif208157TIFF2025122018000305.tif120161 1 Colo320DM cell viability 2 Alpha Screen

[0843] Example 5 - Wnt reporter assay Wnt / beta-catenin-LEF-TCF-bla HCT116-inhibitor screen, constitutively activated. LEF-TCF-bla HCT116 cells were thawed and prepared as described above for the activator screen. 32 μL of cell suspension was added to each well of a 384-well poly-D-lysine assay plate. Cells in assay media were incubated in the plate at 37°C / 5% CO2 in a humidified incubator for 16–24 hours. 4 μL of 10-fold serially diluted ICG-001 (control inhibitor, starting concentration 25,000 nM) or compound was added to the appropriate wells of the plate. 4 μL of assay media was added to all wells for a final assay volume of 40 μL. The plate was incubated for 5 hours in a humidified incubator at 37°C / 5% CO2. 8 μL of 1 μM substrate loading solution was added to each well, and the plate was incubated at room temperature for 2 hours. The plate was read on a fluorescent plate reader. [Table 43]

[0844] Other embodiments Although the present application has been described with its detailed description, it should be understood that the above description is intended to illustrate, but not to limit, the scope of the application, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

[0845] Sequence Listing <110> WNTRX PHARMACEUTICALS INC. <120> BCL9 PEPTIDES AND VARIANTS THEREOF <130> PA25-192 <150> US 62 / 583,820 <151> 2017-11-09 <160> 279 <170> PatentIn version 3.5 <210> 1 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (2)..(2) <223> Leu, Ala, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, Beta-cyclobutyl alanine, D-Leucine, N-methyl leucine, N-methyl cyclohexyl alanine, Cyclohexyl D-alanine or L-alpha-neopentylglycine <220> <221> MOD_RES <222> (5)..(5) <223> Leu, Ala, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, Beta-cyclobutyl alanine, D-Leucine, N-methyl leucine, N -methyl cyclohexyl alanine, Cyclohexyl D-alanine or L- alpha-neopentylglycine <220> <221> MOD_RES <222> (7)..(7) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (8)..(8) <223> Ile, Ala, Norleucine, N-methyl-L-isoleucine, Beta- cyclobutylalanine or D-Isoleucine <220> <221> MOD_RES <222> (10)..(10) <223> Arg, Ala, Gln, Glu, Lys, His, N-methylarginine, Homoarginine, N-methylarginine, Norarginine or Citrulline <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> Leu, Ala, Beta-cyclobutyl alanine, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, 4-Chlorophenyl alanine, D-Leucine, Alpha methylleucine, Cyclohexyl D-alanine, N-methyl cyclohexyl alanine, Allyl glycine, 1-aminocyclobutane-1 <220> <221> MOD_RES <222> (12)..(12) <223> CONT. FROM ABOVE: -carboxylic acid, 1-aminocyclohexane-1- carboxylic acid, Azetidine-3-carboxylic acid, N-methyl cyclohexyl alanine, Beta-tBu-Ala, Tertleucine, 4-fluorophenyl alanine, or 3,4-Chlorophenyl alanine <220> <221> MOD_RES <222> (13)..(13) <223> 2-Naphthylalanine <400> 1 Ser Xaa Gln Thr Xaa Arg Xaa Xaa Gln Xaa Xaa Xaa Ala 1 5 10 <210> 2 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (7)..(7) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (7)..(11) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (13)..(13) <223> 2-Naphthylalanine <400> 2 Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu Ala 1 5 10 <210> 3 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (7)..(7) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (7)..(11) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> Beta-cyclobutyl alanine <220> <221> MOD_RES <222> (13)..(13) <223> 2-Naphthylalanine <400> 3 Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Ala Ala 1 5 10 <210> 4 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (8)..(8) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (8)..(12) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (12)..(12) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (14)..(14) <223> 2-Naphthylalanine <400> 4 Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu Ala 1 5 10 <210> 5 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (10)..(14) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (14)..(14) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (16)..(16) <223> 2-Naphthylalanine <400> 5 Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu Ala 1 5 10 15 <210> 6 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (11)..(15) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (15)..(15) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (17)..(17) <223> 2-Naphthylalanine <400> 6 His Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu 1 5 10 15 Ala <210> 7 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (12)..(12) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (12)..(16) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (16)..(16) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (18)..(18) <223> 2-Naphthylalanine <400> 7 Glu His Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa 1 5 10 15 Leu Ala <210> 8 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (14)..(14) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (14)..(18) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (18)..(18) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (20)..(20) <223> 2-Naphthylalanine <400> 8 Gln Leu Glu His Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln 1 5 10 15 Arg Xaa Leu Ala 20 <210> 9 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (12)..(12) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (12)..(16) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (16)..(16) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (17)..(17) <223> Beta-cyclobutyl alanine <220> <221> MOD_RES <222> (18)..(18) <223> 2-Naphthylalanine <400> 9 Glu His Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa 1 5 10 15 Ala Ala <210> 10 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (10)..(14) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (14)..(14) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (16)..(16) <223> 2-Naphthylalanine <400> 10 Gln Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu Ala 1 5 10 15 <210> 11 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (11)..(15) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (15)..(15) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (17)..(17) <223> 2-Naphthylalanine <400> 11 His Gln Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Leu 1 5 10 15 Ala <210> 12 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (12)..(12) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (12)..(16) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (16)..(16) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (18)..(18) <223> 2-Naphthylalanine <400> 12 Glu His Gln Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa 1 5 10 15 Leu Ala <210> 13 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (10)..(14) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (14)..(14) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (15)..(15) <223> Beta-cyclobutyl alanine <220> <221> MOD_RES <222> (16)..(16) <223> 2-Naphthylalanine <400> 13 Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Ala Ala 1 5 10 15 <210> 14 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (11)..(11) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (11)..(15) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (15)..(15) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (16)..(16) <223> Beta-cyclobutyl alanine <220> <221> MOD_RES <222> (17)..(17) <223> 2-Naphthylalanine <400> 14 His Arg Glu Arg Ser Leu Gln Thr Leu Arg Xaa Ile Gln Arg Xaa Ala 1 5 10 15 Ala <210> 15 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (1)..(1) <223> Leu, Ala, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, Beta-cyclobutyl alanine, D-Leucine, N-methyl leucine, N-methyl cyclohexyl alanine, Cyclohexyl D-alanine or L-alpha-neopentylglycine <220> <221> MOD_RES <222> (4)..(4) <223> Leu, Ala, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, Beta-cyclobutyl alanine, D-Leucine, N-methyl leucine, N -methyl cyclohexyl alanine, Cyclohexyl D-alanine or L- alpha-neopentylglycine <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (7)..(7) <223> Ile, Ala, Norleucine, N-methyl-L-isoleucine, Beta- cyclobutylalanine or D-Isoleucine <220> <221> MOD_RES <222> (9)..(9) <223> Arg, Ala, Gln, Glu, Lys, His, N-methylarginine, Homoarginine, N-methylarginine, Norarginine or Citrulline <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (11)..(11) <223> Leu, Ala, Beta-cyclobutyl alanine, Cyclohexyl L-alanine, Cyclopropyl-L-alanine, 4-Chlorophenyl alanine, D-Leucine, Alpha methylleucine, Cyclohexyl D-alanine, N-methyl cyclohexyl alanine, Allyl glycine, 1-aminocyclobutane-1 <220> <221> MOD_RES <222> (11)..(11) <223> CONT. FROM ABOVE: -carboxylic acid, 1-aminocyclohexane-1- carboxylic acid, Azetidine-3-carboxylic acid, N-methyl cyclohexyl alanine, Beta-tBu-Ala, Tertleucine, 4-fluorophenyl alanine, or 3,4-Chlorophenyl alanine <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 15 Xaa Gln Thr Xaa Arg Xaa Xaa Gln Xaa Xaa Xaa Ala 1 5 10 <210> 16 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (6)..(10) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 16 Ala Gln Thr Ala Arg Xaa Ile Gln Arg Xaa Leu Ala 1 5 10 <210> 17 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (6)..(10) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 17 Leu Gln Thr Ala Arg Xaa Ala Gln Arg Xaa Leu Ala 1 5 10 <210> 18 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (6)..(10) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 18 Leu Gln Thr Leu Arg Xaa Ala Gln Arg Xaa Ala Ala 1 5 10 <210> 19 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (6)..(10) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 19 Leu Gln Thr Leu Arg Xaa Ile Gln Ala Xaa Leu Ala 1 5 10 <210> 20 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <220> <221> MOD_RES <222> (6)..(6) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MISC_FEATURE <222> (6)..(10) <223> May or may not be a hydrocarbon crosslinker between residues <220> <221> MOD_RES <222> (10)..(10) <223> Any alpha, alpha-disubstituted amino acid <220> <221> MOD_RES <222> (11)..(11) <223> Beta-cyclobutyl alanine <220> <221> MOD_RES <222> (12)..(12) <223> 2-Naphthylalanine <400> 20 Leu Gln Thr Leu Arg Xaa Ile Gln Ala Xaa Ala Ala 1 5 10 <210> 21 <211> 12 <212> PRT ...

Claims

1. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: 【Table 1】 During the ceremony, Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; The polypeptide has a length of 6 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

2. Xaa ７ The polypeptide of claim 1, wherein is CBA.

3. Xaa ７ The polypeptide of claim 1 , wherein is selected from Cha and Cpa.

4. Xaa ７ 2. The polypeptide of claim 1, wherein is selected from α-methyl L, DCha, N-methyl Cha, and allyl Gly.

5. Xaa ７ 2. The polypeptide of claim 1, wherein is selected from AC4C, A6C, Aze, Phe(4-Cl), (β-tBu-Ala), and Tle.

6. Xaa ７ The polypeptide of claim 1, wherein is selected from Phe(4-Cl), 4-FPh, 3,4-diClPh, and Cha.

7. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1, wherein said amino acid sequence comprises an IQR.

8. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1, wherein said polypeptide comprises I(N-methylQ)R.

9. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1, wherein said amino acid sequence is IQ(N-methyl R).

10. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1 , wherein the amino acid sequence comprises (CBA)QR.

11. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1, wherein said amino acid sequence is IQ (homo R).

12. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 1, wherein said amino acid sequence comprises (N-methyl I)QR.

13. Xaa ４ Xaa ８ Xaa ５ 2. The polypeptide of claim 1, wherein said amino acid sequence comprises IQQ, IQE, IQ(NMeArg), (Nle)QR, IQ(Nar), or IQCit.

14. the polypeptide comprising: 【Table 2】 During the ceremony, Xaa １ and Xaa ２ are each independently selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa ９ is selected from Q, E, N-methyl Q, N-MeGln, and peptoid Q; Xaa １０ is selected from T, N-methyl T, and DThr; Xaa １１ The polypeptide of any one of claims 1 to 13, wherein is selected from R, N-methylR, E, K, homoR, Nar, and Cit.

15. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ The polypeptide of claim 14 , wherein said polypeptide comprises LQTLR.

16. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ The polypeptide of claim 14, wherein said polypeptide comprises an L(N-methyl Q) TLR.

17. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ The polypeptide of claim 14, wherein said amino acid sequence comprises L(N-methylQ)TL(homoR).

18. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ The polypeptide of claim 14, wherein said amino acid sequence comprises L(NMeGln)T(NMeLeu)R.

19. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ The polypeptide of claim 14, wherein said amino acid sequence comprises L(N-methyl Q)TL(N-methyl R).

20. Xaa １ Xaa ９ Xaa １０ Xaa ２ Xaa １１ 15. The polypeptide of claim 14, wherein said polypeptide comprises LN-methyl QTLR, LETLR, (CBA)QTLR, (CBA)(N-methyl Q)TLR, LQT(CBA)R, L(N-methyl Q)(N-methyl T)LR, L(N-methyl Q)T(Cha)R, L(N-methyl Q)T(α-methyl L)R, or L(N-methyl Q)(DThr)LR.

21. the polypeptide comprising: 【Table 3】 During the ceremony, Xaa １２ is selected from H, N-MeHis, Cys, N-MeCys, homoHis, and NHis; Xaa １３ is selected from R, N-methylR, HomoArg, Cit, Nar, and Phe(4-guanidino); Xaa １４ is selected from E, Q, N-methyl E, N-methyl Q, N-methyl D, and NMeGln; Xaa １５ is selected from R, homoR, and N-methylR.

22. Xaa １２ Xaa １３ Xaa １４ Xaa １５ The polypeptide of claim 21 , wherein said polypeptide comprises an HRER.

23. Xaa １２ Xaa １３ Xaa １４ Xaa １５ The polypeptide of claim 21 , wherein the sequence comprises HRQR.

24. Xaa １２ Xaa １３ Xaa １４ Xaa １５ comprises HR(N-methyl E)R, HR(N-methyl E)R, HR(N-methyl Q)R, HR(N-methyl D)R, H(N-methyl R)QR, HRQ(Homo R), HRQ(N-methyl R), H(Homo Arg)QR, HRQ(NMeArg), HR(NMeGln)R, (N-MeHis)RQR, (Cys)RQR, (NMeCys)RQR, (Homo His)RQR, (NHis)RQR, H(Cit)(N-methyl Q)R, H(Nar)(N-methyl Q)R, or H(4-guanidino-Phe)(N-methyl Q)R.

25. The N-terminus of the polypeptide is acetyl, propionyl, hexanoyl, 3-phenylpropanoyl, 2-cyclohexylacetyl, diphenylacetyl, 3,5-dihydroxybenzoic acid, 4-(trifluoromethyl)benzoic acid, 5-phenylvaleric acid, 4-biphenylacetic acid, dimethyl, HOCH ２ CH ２ The polypeptide of any one of claims 1 to 24, which is modified with a moiety selected from CO- and palmitoyl-PEG4.

26. The C-terminus of the polypeptide is NH ２ , (β-Ala) (β-Ala), (β-Ala) (β-Ala)NH ２ , GRKKRRQRRRPQK(PEG4-palmitoyl)NH ２ ,GRKKRRQRRRPQNH ２ , K(PEG4-palmitoyl)NH ２ 26. The polypeptide of any one of claims 1 to 25, modified with a moiety selected from the group consisting of:

27. The C-terminus of the polypeptide is NH ２ 27. The polypeptide of claim 26, modified with

28. 27. The polypeptide of claim 26, wherein the C-terminus of the polypeptide is modified with (β-Ala)(β-Ala).

29. The N-terminus of the polypeptide is modified with acetyl and the C-terminus of the polypeptide is modified with NH ２ The polypeptide according to any one of claims 1 to 24, which is modified with

30. The polypeptide of claim 1 , wherein the polypeptide is selected from the following: 【Table 4】

31. The polypeptide of any one of claims 1 to 30, wherein the α,α-disubstituted amino acid is an α-methyl,α-alkenyl amino acid.

32. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

33. Xaa ３ and Xaa ６ and each is (S)-2-(4'-pentenyl)alanine.

34. 31. The polypeptide of any one of claims 1 to 30, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

35. The hydrocarbon linker has the formula: 【Chemical 1】 In the formula, one 【Chemistry 2】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 3】 is Xaa of the hydrocarbon linker ６ 35. The polypeptide of claim 34, wherein the point of attachment to the alpha carbon atom is:

36. 36. The polypeptide of claim 35, wherein the hydrocarbon crosslinker has the formula: 【Chemistry 4】

37. A polypeptide, or a pharmaceutically acceptable salt thereof, having the amino acid sequence: 【Table 5】 an amino acid sequence selected from: 【Table 6】 During the ceremony, Xaa ３ , Xaa ６ , Xaa １６ , and Xaa １０ are each independently an α,α-disubstituted amino acid; Xaa １ is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa ９ is selected from Q, N-methyl Q, E, N-MeGln, and peptoid Q; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; The polypeptide has a length of 9 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

38. Xaa １ 38. The polypeptide of claim 37, wherein is selected from L and Cpa.

39. Xaa ９ is selected from Q and N-methyl Q.

40. Xaa ４ The polypeptide of claim 37, wherein is I.

41. Xaa ８ is Q.

42. Xaa ５ is R.

43. Xaa １ Xaa ９ The polypeptide of claim 37, wherein comprises LQ, L(N-methylQ), or (Cpa)(N-methylQ).

44. Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 37, wherein comprises an IQR.

45. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 37 to 44, comprising at least one R in addition to

46. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 37 to 44, comprising at least one L in addition to

47. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ 45. The polypeptide of claim 37, further comprising at least one amino acid selected from CBA, Cpa, and Cha.

48. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 37 to 44, further comprising at least one (2-Nal).

49. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 37 to 44, further comprising at least one (β-Ala).

50. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 37 to 49, further comprising HRQR or HRER.

51. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 37 to 50, further comprising LR, (Cpa)R, or (Cha)R.

52. The polypeptide has the sequence Xaa １６ Xaa １ Xaa ９ Xaa １０ and Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 37 to 51, further comprising (CBA)(2-Nal), (Cpa)(2-Nal), or (Cha)(2-Nal).

53. 38. The polypeptide of claim 37, wherein the polypeptide is selected from the following: RXaa １６ L(N-methylQ)Xaa １０ LRXaa ３ IQRXaa ６ (CBA)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 88), RXaa １６ (Cpa)(N-methylQ)Xaa １０ (Cpa)RXaa ３ IQRXaa ６ (Cpa)(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 89), HRQRXaa １６ LQXaa １０ LRXaa ３ IQRXaa ６ (CBA)(2-Nal) (SEQ ID NO: 90), HRQRXaa １６ LQXaa １０ (Cpa)RXaa ３ IQRXaa ６ (Cpa)(2-Nal) (SEQ ID NO: 91), HRQRXaa １６ LQXaa １０ (Cha)RXaa ３ IQRXaa ６ (Cha)(2-Nal) (SEQ ID NO: 92), and LEHRERXaa １６ LQXaa １０ LRXaa ３ IQRXaa ６ L (sequence number 93).

54. 54. The polypeptide of any one of claims 37 to 53, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

55. Xaa ３ , Xaa ６ , Xaa １０ , and Xaa １６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

56. Xaa ３ , Xaa ６ , Xaa １０ , and Xaa １６ and each is (S)-2-(4'-pentenyl)alanine.

57. The α,α-disubstituted amino acid Xaa ３ one α-substituent in the acid is methyl and the other α-substituent is a first hydrocarbon linker, and said α,α-disubstituted amino acid Xaa ６ wherein one alpha substituent is methyl and the other alpha substituent is said first hydrocarbon linker; The α,α-disubstituted amino acid Xaa １０ one α-substituent in the acid is methyl and the other α-substituent is a second hydrocarbon linker, and said α,α-disubstituted amino acid Xaa １６ 54. The polypeptide of any one of claims 37 to 53, wherein one alpha substituent is methyl and the other alpha substituent is the second hydrocarbon linker.

58. The first hydrocarbon linker has the formula: 【Chemistry 5】 In the formula, one 【Chemistry 6】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 7】 is Xaa of the hydrocarbon linker ６ indicates the point of attachment to the α carbon atom of the second hydrocarbon linker has the formula: 【Chemistry 8】 In the formula, one 【Chemistry 9】 is Xaa of the second hydrocarbon linker １０ indicates the point of attachment to the α carbon atom of 【Chemistry 10】 is Xaa of the second hydrocarbon linker １６ 58. The polypeptide of claim 57, wherein the point of attachment to the alpha carbon atom is:

59. the first hydrocarbon crosslinker having the formula: 【Chemistry 11】 59. The polypeptide of claim 58, wherein the second hydrocarbon crosslinker has the formula: 【Chemistry 12】

60. A polypeptide, or a pharmaceutically acceptable salt thereof, having the amino acid sequence: 【Table 7】 an amino acid sequence selected from: 【Table 8】 During the ceremony, Xaa ３ , Xaa ６ , Xaa ９ , and Xaa １４ are each independently an α,α-disubstituted amino acid; Xaa １５ is selected from R, homoR, and N-methylR; Xaa １６ is selected from S and T; Xaa １ is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; The polypeptide has a length of 9 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

61. Xaa １５ is R.

62. Xaa １６ is S.

63. Xaa １ is L.

64. Xaa １５ Xaa １６ Xaa １ The polypeptide of claim 60, wherein said polypeptide comprises an RSL.

65. Xaa ４ The polypeptide of any one of claims 60 to 64, wherein is I.

66. Xaa ８ The polypeptide of any one of claims 60 to 64, wherein is Q.

67. Xaa ５ The polypeptide of any one of claims 60 to 64, wherein is R.

68. Xaa ４ Xaa ８ Xaa ５ The polypeptide of any one of claims 60 to 64, wherein comprises IQR.

69. The polypeptide has the amino acid sequence Xaa １４ Xaa １５ Xaa １６ Xaa １ Xaa ９ and the amino acid sequence Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ 69. The polypeptide of any one of claims 60 to 68, comprising at least one R in addition to

70. The polypeptide has the amino acid sequence Xaa １４ Xaa １５ Xaa １６ Xaa １ Xaa ９ and the amino acid sequence Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 60 to 68, further comprising at least one (2-Nal).

71. The polypeptide has the amino acid sequence Xaa １４ Xaa １５ Xaa １６ Xaa １ Xaa ９ and the amino acid sequence Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 60 to 70, comprising HR in addition to

72. The polypeptide has the amino acid sequence Xaa １４ Xaa １５ Xaa １６ Xaa １ Xaa ９ and the amino acid sequence Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 60 to 70, comprising a TLR in addition to the above.

73. The polypeptide has the amino acid sequence Xaa １４ Xaa １５ Xaa １６ Xaa １ Xaa ９ and the amino acid sequence Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 60 to 70, further comprising (CBA)(2-Nal) or (4-ClPh)(2-Nal).

74. The polypeptide is selected from: HRXaa １４ RSLXaa ９ TLRXaa ３ IQRXaa ６ (CBA)(2-Nal) (SEQ ID NO: 94), HRXaa １４ RSLXaa ９ TLRXaa ３ IQRXaa ６ (CBA)(2-Nal) (SEQ ID NO: 95), HRXaa １４ RSLXaa ９ TLRXaa ３ IQRXaa ６ (4-ClPh)(2-Nal) (SEQ ID NO: 96), and HRXaa １４ RSLXaa ９ TLRXaa ３ IQRXaa ６ (4-Cl-Ph)(2-Nal) (SEQ ID NO: 198), The polypeptide of claim 60, wherein the N-terminus of SEQ ID NO: 95 and SEQ ID NO: 96 is modified with palmitoyl-PEG4.

75. 75. The polypeptide of any one of claims 60 to 74, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

76. Xaa ３ , Xaa ６ , Xaa ９ , and Xaa １４ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

77. Xaa ３ , Xaa ６ , Xaa ９ , and Xaa １４ and each is (S)-2-(4'-pentenyl)alanine.

78. The α,α-disubstituted amino acid Xaa ３ one α-substituent in the acid is methyl and the other α-substituent is a first hydrocarbon linker, and said α,α-disubstituted amino acid Xaa ６ wherein one alpha substituent is methyl and the other alpha substituent is said first hydrocarbon linker; The α,α-disubstituted amino acid Xaa ９ one α-substituent in the acid is methyl and the other α-substituent is a second hydrocarbon linker, and said α,α-disubstituted amino acid Xaa １４ 75. The polypeptide of any one of claims 60 to 74, wherein one alpha substituent is methyl and the other alpha substituent is the second hydrocarbon linker.

79. The first hydrocarbon linker has the formula: 【Chemistry 13】 In the formula, one 【Chemistry 14】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 15】 is Xaa of the hydrocarbon linker ６ indicates the point of attachment to the α carbon atom of the second hydrocarbon linker has the formula: 【Chemistry 16】 In the formula, one 【Chemistry 17】 is Xaa of the second hydrocarbon linker ９ indicates the point of attachment to the α carbon atom of 【Chemistry 18】 is Xaa of the second hydrocarbon linker １４ indicates the point of attachment to the α carbon atom of 79. The polypeptide of claim 78.

80. the first hydrocarbon crosslinker having the formula: 【Chemistry 19】 80. The polypeptide of claim 79, wherein the second hydrocarbon crosslinker has the formula: 【Chemistry 20】

81. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: 【Table 9】 Xaa １０ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ２ is selected from L, A, Cha, Cpa, (DL), CBA, MeL, NMeCha, Dcha, and NptGly; Xaa １１ is selected from R, N-methylR, E, K, homoR, Nar, and Cit; Xaa ３ is selected from D and Nle; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; The polypeptide has a length of 8 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

82. Xaa ２ is L.

83. Xaa １１ is R.

84. Xaa ３ is D.

85. Xaa ４ The polypeptide of claim 81, wherein is I.

86. Xaa ８ is Q.

87. Xaa ５ is R.

88. Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ The polypeptide of claim 81, wherein said polypeptide comprises LRDIQR.

89. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 81 to 88, comprising at least one L in addition to

90. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 81 to 89, further comprising at least one (2-Nal).

91. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 81 to 90, further comprising at least one (β-Ala).

92. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 81 to 91, comprising, in addition to LQ.

93. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide according to any one of claims 81 to 92, further comprising L(2-Nal).

94. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ The polypeptide of any one of claims 81 to 93, further comprising at least one (β-Ala).

95. The polypeptide has the amino acid sequence Xaa １０ Xaa ２ Xaa １１ Xaa ３ Xaa ４ Xaa ８ Xaa ５ Xaa ６ 95. The polypeptide of any one of claims 81 to 94, comprising HRERS or HRQRS in addition to:

96. the polypeptide LQXaa １０ LRDIQRXaa ６ 82. The polypeptide of claim 81, wherein the polypeptide is L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 97).

97. 97. The polypeptide of any one of claims 81 to 96, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

98. Xaa ６ and Xaa １０ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

99. Xaa ６ and Xaa １０ and each is (S)-2-(4'-pentenyl)alanine.

100. 97. The polypeptide of any one of claims 81 to 96, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

101. The hydrocarbon linker has the formula: 【Chemical 21】 In the formula, one 【Chemical formula 22】 is Xaa of the hydrocarbon linker ６ indicates the point of attachment to the α carbon atom of 【Chemical 23】 is Xaa of the hydrocarbon linker １０ 101. The polypeptide of claim 100, wherein the point of attachment to the alpha carbon atom is:

102. 102. The polypeptide of claim 101, wherein the hydrocarbon crosslinker has the formula: 【Chemistry 24】

103. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: １ QTXaa ２ RXaa ３ Xaa ４ QXaa ５ Xaa ６ Xaa ７ (2-Nal) (SEQ ID NO: 1), During the ceremony, Xaa １ and Xaa ２ are each independently selected from L, A, Cha, Cpa, CBA, (DL), MeL, NMeCha, Dcha, and NptGly; Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; A polypeptide, or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a length of 13 to 30 amino acids.

104. Xaa １ and Xaa ２ are each independently L or A; Xaa ４ is I or A, Xaa ５ is R or A, Xaa ７ is L, A, or CBA.

105. Xaa １ and Xaa ２ and each is L.

106. Xaa １ and Xaa ２ The polypeptide of claim 103, wherein each of

107. Xaa １ is L, and Xaa ２ The polypeptide of claim 103, wherein is A.

108. Xaa １ and Xaa ２ and each is Cha.

109. Xaa １ and Xaa ２ The polypeptide of claim 103, wherein each of

110. Xaa ４ is I, and Xaa ５ The polypeptide of any one of claims 103 to 109, wherein is R.

111. Xaa ４ is I, and Xaa ５ The polypeptide of any one of claims 103 to 109, wherein is A.

112. Xaa ４ is A, and Xaa ５ The polypeptide of any one of claims 103 to 109, wherein is R.

113. Xaa ７ is selected from Cha, Cpa, Phe(4-Cl), (DL), α-methyl L, DCcha, N-methylcha, allylGly, AC4C, A6C, Aze, N-Mecha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh.

114. Xaa ７ The polypeptide according to any one of claims 103 to 112, wherein is L.

115. Xaa ７ The polypeptide according to any one of claims 103 to 112, wherein is A.

116. Xaa ７ The polypeptide according to any one of claims 103 to 112, wherein is CBA.

117. The amino acid sequence SEQ ID NO:1 is selected from: SLQTLRXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 2), and SLQTLRXaa ３ IQRXaa ６ (CBA)(2-Nal) (SEQ ID NO: 3), the polypeptide of claim 103.

118. 118. The polypeptide of any one of claims 103 to 117, wherein the polypeptide comprises the amino acid sequence RER.

119. The polypeptide of any one of claims 103 to 117, wherein the polypeptide comprises the amino acid sequence QER.

120. 120. The polypeptide of any one of claims 103 to 119, wherein the polypeptide has a length of 13 to 20 amino acids.

121. 121. The polypeptide of any one of claims 103 to 120, wherein the polypeptide comprises the amino acid sequence SEQ ID NO: 1 plus at least one amino acid selected from Q, L, E, H, and R.

122. The polypeptide is selected from any one of the following polypeptides: 【Table 10】 The polypeptide of claim 103.

123. 123. The polypeptide of any one of claims 103 to 122, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

124. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

125. Xaa ３ and Xaa ６ and each is (S)-2-(4'-pentenyl)alanine.

126. 124. The polypeptide of any one of claims 103 to 123, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

127. The hydrocarbon linker has the formula: 【Chemistry 25】 In the formula, one 【Chemical 26】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemical 27】 is Xaa of the hydrocarbon linker ６ 127. The polypeptide of claim 126, wherein the point of attachment to the alpha carbon atom is:

128. 128. The polypeptide of claim 127, wherein the hydrocarbon crosslinker has the formula: 【Chemical Formula 28】

129. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: １ QTXaa ２ RXaa ３ Xaa ４ QXaa ５ Xaa ６ Xaa ７ (2-Nal) (SEQ ID NO: 15), During the ceremony, Xaa １ and Xaa ２ are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; the polypeptide comprises at least one A; The polypeptide has a length of 12 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

130. Xaa １ and Xaa ２ are each independently L or A; Xaa ４ is I or A, Xaa ５ is R or A, Xaa ７ is L, A, or CBA.

131. Xaa １ , Xaa ２ , Xaa ４ , Xaa ５ , and Xaa ７ 130. The polypeptide of claim 129, wherein at least one of is A.

132. Xaa １ and Xaa ２ and each is A.

133. Xaa １ is L, and Xaa ２ The polypeptide of claim 129, wherein is A.

134. Xaa １ and Xaa ２ and each is L.

135. Xaa ２ and Xaa ４ and each is A.

136. Xaa ２ is L, and Xaa ４ The polypeptide of any one of claims 129 to 131, wherein is A.

137. Xaa ２ is L, and Xaa ４ The polypeptide of any one of claims 129 to 131, wherein

138. Xaa ２ is A, and Xaa ４ The polypeptide of any one of claims 129 to 131, wherein

139. Xaa ４ is I, and Xaa ７ The polypeptide of any one of claims 129 to 131, wherein is A.

140. Xaa ５ is R, and Xaa ７ is L.

141. Xaa ５ is R, and Xaa ７ The polypeptide of any one of claims 129 to 139, wherein is A.

142. Xaa ５ is A, and Xaa ７ is L.

143. Xaa ５ is R, and Xaa ７ The polypeptide of any one of claims 129 to 139, wherein is CBA.

144. Xaa ５ is A, and Xaa ７ The polypeptide of any one of claims 129 to 139, wherein is CBA.

145. The amino acid sequence SEQ ID NO:15 is selected from: AQTARXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 16), LQTARXaa ３ AQRXaa ６ L(2-Nal) (SEQ ID NO: 17), LQTLRXaa ３ AQRXaa ６ A(2-Nal) (SEQ ID NO: 18), LQTLRXaa ３ IQAXaa ６ L(2-Nal) (SEQ ID NO: 19), and LQTLRXaa ３ IQAXaa ６ (CBA)(2-Nal) (SEQ ID NO: 20), the polypeptide of claim 129.

146. 146. The polypeptide of any one of claims 129 to 145, wherein the polypeptide comprises the amino acid sequence ERS.

147. 147. The polypeptide of any one of claims 129 to 146, wherein the polypeptide comprises the amino acid sequence (β-Ala)(β-Ala).

148. 148. The polypeptide of any one of claims 129 to 147, wherein the polypeptide comprises the amino acid sequence AA.

149. 149. The polypeptide of any one of claims 129 to 148, wherein the polypeptide has a length of 12 to 20 amino acids.

150. 150. The polypeptide of any one of claims 129 to 149, wherein the polypeptide comprises the amino acid sequence SEQ ID NO: 15 plus at least one amino acid selected from Q, L, E, H, R, and S.

151. The polypeptide is selected from any one of the following polypeptides: AQTARXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 21), LQTARXaa ３ AQRXaa ６ L(2-Nal) (SEQ ID NO: 22), LQTLRXaa ３ AQRXaa ６ A(2-Nal) (SEQ ID NO: 23), LQTLRXaa ３ IQAXaa ６ L(2-Nal) (SEQ ID NO: 24), LQTLRXaa ３ IQAXaa ６ (CBA)(2-Nal) (SEQ ID NO: 25), ______________________ ３ |!!| ６ (*)()(β)(|) (β)(4) LQTLRXaa ３ IQAXaa ６ L(2-Nal)AA (SEQ ID NO: 27), HRERSLQTLRXaa ３ IQAXaa ６ L(2-Nal) (SEQ ID NO: 28), HRERSLQTLRXaa ３ IQAXaa ６ (CBA)(2-Nal) (SEQ ID NO: 29), LQTARXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 75), LQTLRXaa ３ AQRXaa ６ L(2-Nal) (SEQ ID NO: 76), and LQTLRXaa ３ IQRXaa ６ A(2-Nal) (SEQ ID NO: 77), The polypeptide of claim 129.

152. The polypeptide is selected from any one of the following polypeptides: AQTARXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 21), LQTARXaa ３ AQRXaa ６ L(2-Nal) (SEQ ID NO: 22), LQTLRXaa ３ AQRXaa ６ A(2-Nal) (SEQ ID NO: 23), LQTLRXaa ３ IQAXaa ６ L(2-Nal) (SEQ ID NO: 24), LQTLRXaa ３ IQAXaa ６ (CBA)(2-Nal) (SEQ ID NO: 25), ______________________ ３ |!!| ６ (*)()(β)(|) (β)(4) LQTLRXaa ３ IQAXaa ６ L(2-Nal)AA (SEQ ID NO: 27), HRERSLQTLRXaa ３ IQAXaa ６ L(2-Nal) (SEQ ID NO: 28), and HRERSLQTLRXaa ３ IQAXaa ６ (CBA)(2-Nal) (SEQ ID NO: 29), The polypeptide of claim 129.

153. 153. The polypeptide of any one of claims 129 to 152, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

154. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

155. Xaa ３ and Xaa ６ is each (S)-2-(4'-pentenyl)alanine.

156. 153. The polypeptide of any one of claims 129 to 152, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

157. The hydrocarbon linker has the formula: 【Chemical Formula 29】 In the formula, one 【Chemistry 30】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemical 31】 is Xaa of the hydrocarbon linker ６ 157. The polypeptide of claim 156, wherein the point of attachment to the alpha carbon atom is:

158. 158. The polypeptide of claim 157, wherein the hydrocarbon linker has the following formula: 【Chemical 32】

159. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: １ QTXaa ２ RXaa ３ Xaa ４ QXaa ５ Xaa ６ Xaa ７ (2-Nal) (SEQ ID NO: 30), During the ceremony, Xaa １ and Xaa ２ are each independently L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, N-MeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; A polypeptide, or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a length of 13 to 22 amino acids.

160. Xaa １ and Xaa ２ are each independently L or A; Xaa ４ is I or A, Xaa ５ is R or A, Xaa ７ is L, A, or CBA.

161. Xaa １ and Xaa ２ and each is L.

162. Xaa ４ is I, and Xaa ５ is R.

163. Xaa ７ is L.

164. The amino acid sequence SEQ ID NO: 30 is LQTLRXaa ３ IQRXaa ６ 160. The polypeptide of claim 159, wherein the polypeptide is L(2-Nal) (SEQ ID NO: 31).

165. 165. The polypeptide of any one of claims 159 to 164, wherein the polypeptide comprises the amino acid sequence SEQ ID NO: 30 plus at least one amino acid selected from P, D, and β-Ala.

166. 166. The polypeptide of any one of claims 159 to 165, wherein the polypeptide comprises the amino acid sequence SEQ ID NO: 30 plus at least one amino acid selected from Q, L, E, H, R, and S.

167. The polypeptide is selected from any one of the following polypeptides: LQTLRXaa ３ IQRXaa ６ L(2-Nal)PD (SEQ ID NO: 32), LQTLRXaa ３ IQRXaa ６ L(2-Nal)P (SEQ ID NO: 33), LQTLRXaa ３ IQRXaa ６ L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34), and LQTLRXaa ３ IQRXaa ６ L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34a), wherein the C-terminus of SEQ ID NO: 34a is GRKKRRQRRRPQK(PEG4-palmitoyl)NH ２ 160. The polypeptide of claim 159, modified with

168. The polypeptide is selected from any one of the following polypeptides: LQTLRXaa ３ IQRXaa ６ L(2-Nal)PD (SEQ ID NO: 32), LQTLRXaa ３ IQRXaa ６ L(2-Nal)P (SEQ ID NO: 33), and LQTLRXaa ３ IQRXaa ６ L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 34), The polypeptide of claim 159.

169. 169. The polypeptide of any one of claims 159 to 168, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

170. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

171. Xaa ３ and Xaa ６ is each (S)-2-(4'-pentenyl)alanine.

172. 169. The polypeptide of any one of claims 159 to 168, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

173. The hydrocarbon linker has the formula: 【Chemical Formula 33】 In the formula, one 【Chemical Formula 34】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 35】 is Xaa of the hydrocarbon linker ６ 173. The polypeptide of claim 172, wherein the point of attachment to the alpha carbon atom is:

174. 174. The polypeptide of claim 173, wherein the hydrocarbon crosslinker has the formula: 【Chemical 36】

175. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: １ TXaa ２ RXaa ３ (SEQ ID NO: 35), During the ceremony, Xaa １ and Xaa ３ are each independently an α,α-disubstituted amino acid; Xaa ２ is selected from L, A, Cha, Cpa, (DL), CBA, MeL, N-MeCha, Dcha, and NptGly; the polypeptide comprises at least one 2-Nal; The polypeptide has a length of 6 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

176. Xaa ２ is L or A.

177. Xaa ２ is L.

178. 178. The polypeptide of any one of claims 175-177, wherein the polypeptide comprises the amino acid sequence SEQ ID NO:35 plus at least one amino acid selected from Q, L, E, H, I, S, M, and R.

179. 176. The polypeptide of claim 175, wherein the polypeptide comprises at least two R's in addition to the amino acid sequence SEQ ID NO:

35.

180. 176. The polypeptide of claim 175, wherein the polypeptide comprises three Rs in addition to the amino acid sequence of SEQ ID NO:

35.

181. 181. The polypeptide of any one of claims 175 to 180, wherein the polypeptide comprises at least two E's in addition to the amino acid sequence SEQ ID NO:

35.

182. 182. The polypeptide of any one of claims 175 to 181, wherein the polypeptide comprises at least two Ls in addition to the amino acid sequence SEQ ID NO:

35.

183. 183. The polypeptide of claim 182, wherein the polypeptide comprises three Ls in addition to the amino acid sequence of SEQ ID NO:

35.

184. 184. The polypeptide of any one of claims 175 to 183, wherein the polypeptide comprises at least two Q's in addition to the amino acid sequence SEQ ID NO:

35.

185. 185. The polypeptide of any one of claims 175 to 184, wherein the polypeptide comprises the amino acid sequence SEQ ID NO: 35 plus at least one amino acid selected from H, S, I, and M.

186. 186. The polypeptide of any one of claims 175 to 185, wherein the polypeptide comprises the amino acid sequence IQR.

187. 187. The polypeptide of any one of claims 175 to 186, wherein the polypeptide comprises the amino acid sequence ML(2-Nal).

188. 188. The polypeptide of any one of claims 175 to 187, wherein the polypeptide comprises the amino acid sequence (2-Abu)L(2-Na1)(β-Ala)(β-Ala).

189. 189. The polypeptide of any one of claims 175 to 188, wherein the polypeptide comprises the amino acid sequence RERSL.

190. 190. The polypeptide of any one of claims 175 to 189, wherein the polypeptide comprises the amino acid sequence QLEH.

191. The polypeptide is selected from: QLEHRERSLXaa １ TLRXaa ３ IQRML(2-Nal) (SEQ ID NO: 36), and QLEHRERSLXaa １ TLRXaa ３ IQR(2-Abu)L(2-Nal)(β-Ala)(β-Ala) (SEQ ID NO: 78), the polypeptide of claim 175.

192. the polypeptide QLEHRERSLXaa １ TLRXaa ３ The polypeptide of claim 175, which is IQRML(2-Nal) (SEQ ID NO: 36).

193. 193. The polypeptide of any one of claims 175 to 192, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

194. Xaa １ and Xaa ３ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

195. Xaa １ and Xaa ３ is each (S)-2-(4'-pentenyl)alanine.

196. 193. The polypeptide of any one of claims 175 to 192, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

197. The hydrocarbon linker has the formula: 【Chemical 37】 In the formula, one 【Chemical Formula 38】 is Xaa of the hydrocarbon linker １ indicates the point of attachment to the α carbon atom of 【Chemical 39】 is Xaa of the hydrocarbon linker ３ 197. The polypeptide of claim 196, wherein the point of attachment to the alpha carbon atom is:

198. 198. The polypeptide of claim 197, wherein the hydrocarbon crosslinker has the formula: 【Chemistry 40】

199. 1. A polypeptide, or a pharmaceutically acceptable salt thereof, having an amino acid sequence selected from: Xaa １ SLQXaa ２ (SEQ ID NO: 37a) and Xaa １ S(Cha)(N-methylQ)Xaa ２ (SEQ ID NO: 37b), and An amino acid sequence selected from: Xaa ３ IQRXaa ４ (SEQ ID NO: 38a) and Xaa ３ IQQXaa ４ (SEQ ID NO: 38b), During the ceremony, Xaa １ , Xaa ２ , Xaa ３ , and Xaa ４ are each independently an α,α-disubstituted amino acid; The polypeptide has a length of 10 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

200. Amino acid sequence: Xaa １ SLQXaa ２ (SEQ ID NO: 37), and Amino acid sequence: Xaa ３ IQRXaa ４ (SEQ ID NO: 38) 200. The polypeptide of claim 199, comprising:

201. 200. The polypeptide of claim 199, wherein the polypeptide comprises at least one 2-Nal.

202. 200. The polypeptide of claim 199, wherein the polypeptide comprises at least one CBA.

203. The polypeptide comprises any one of the amino acid sequences of SEQ ID NO:37a or SEQ ID NO:37b, and the amino acid sequence of SEQ ID NO:38a or SEQ ID NO:38b, as well as L, E, R, H, Q, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, HomoHis, NHis, HomoR, Cit, Nar, Ph 200. The polypeptide of claim 199, comprising at least one amino acid selected from e(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allyl Gly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

204. 200. The polypeptide of claim 199, wherein the polypeptide comprises the amino acid sequence SEQ ID NO:37a or SEQ ID NO:37b, and the amino acid sequence SEQ ID NO:38a or SEQ ID NO:38b, as well as at least one amino acid selected from L, E, R, H, Q, N-methyl E, CBA, N-methyl Q, Cha, and N-methyl R.

205. 205. The polypeptide of any one of claims 199 to 204, wherein the polypeptide comprises the amino acid sequence SEQ ID NO:37 and the amino acid sequence SEQ ID NO:38, plus at least one amino acid selected from L, E, and R.

206. The polypeptide of any one of claims 199 to 205, wherein the polypeptide comprises the amino acid sequence RE.

207. 207. The polypeptide of any one of claims 199 to 206, wherein the polypeptide comprises the amino acid sequence LR.

208. 207. The polypeptide of any one of claims 199 to 206, wherein the polypeptide comprises the amino acid sequence L(2-Nal).

209. 209. The polypeptide of any one of claims 199 to 208, wherein the polypeptide comprises the amino acid sequence (CBA)(2-Nal).

210. 210. The polypeptide of any one of claims 199-209, wherein the polypeptide comprises an amino acid sequence selected from HRE, HR(N-methyl E), HR(N-methyl Q), HRQ, LR, L(N-methyl R), (Cha)R, L(2-Nal), and (CBA)(2-Nal).

211. 211. The polypeptide of any one of claims 199 to 210, wherein the polypeptide comprises at least one β-Ala.

212. 212. The polypeptide of any one of claims 199-211, wherein the polypeptide has a length of 10-20 amino acids.

213. The polypeptide is selected from any one of the following polypeptides: REXaa １ SLQXaa ２ LRXaa ３ IQRXaa ４ L(2-Nal) (SEQ ID NO: 39), ____________ １ _________________ ２ _________________ ３ |!!| ４ (*)()(β)(|) (β)(_)( .. １ _________________ ２ ________________ ３ |!!| ４ (*)()(β)(|) (β)(()) HREXaa １ SLQXaa ２ LRXaa ３ IQRXaa ４ (CBA)(2-Nal) (SEQ ID NO: 82), HREXaa １ SLQXaa ２ LRXaa ３ IQQXaa ４ (CBA)(2-Nal)(SEQ ID NO: 83), HR(N-methyl E)Xaa １ SLQXaa ２ LRXaa ３ IQRXaa ４ (CBA)(2-Nal) (SEQ ID NO: 84), HREXaa １ SLQXaa ２ L(N-methylR)Xaa ３ IQRXaa ４ (CBA)(2-Nal) (SEQ ID NO: 85), HR(N-methylQ)Xaa １ S(Cha)(N-methylQ)Xaa ２ (Cha)RXaa ３ IQRXaa ４ (Cha)(2-Nal) (SEQ ID NO: 86), and HRQXaa １ SLQXaa ２ LRXaa ３ IQRXaa ４ (CBA)(2-Nal)(SEQ ID NO: 87), 200. The polypeptide of claim 199.

214. The polypeptide is selected from any one of the following polypeptides: REXaa １ SLQXaa ２ LRXaa ３ IQRXaa ４ L(2-Nal) (SEQ ID NO: 39), and ____________ １ _________________ ２ _________________ ３ |!!| ４ (*)()(β)(|) (β)(_)( 200. The polypeptide of claim 199.

215. 215. The polypeptide of any one of claims 199 to 214, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

216. Xaa １ , Xaa ２ , Xaa ３ , and Xaa ４ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

217. Xaa １ and Xaa ２ is each (S)-2-(4'-pentenyl)alanine.

218. Xaa ３ and Xaa ４ and each is (S)-2-(4'-pentenyl)alanine.

219. The α,α-disubstituted amino acid Xaa １ wherein one α-substituent is methyl and the other α-substituent is a first hydrocarbon linker, and ２ 219. The polypeptide of any one of claims 199 to 218, wherein one alpha substituent is methyl and the other alpha substituent is the first hydrocarbon linker.

220. The first hydrocarbon linker has the formula: 【Chemistry 41】 In the formula, one 【Chemistry 42】 is Xaa of the first hydrocarbon linker １ indicates the point of attachment to the α carbon atom of 【Chemistry 43】 is Xaa of the first hydrocarbon linker ２ 220. The polypeptide of claim 219, wherein the point of attachment to the alpha carbon atom is:

221. 221. The polypeptide of claim 220, wherein the first hydrocarbon crosslinker has the formula: 【Chemical 44】

222. The α,α-disubstituted amino acid Xaa ３ wherein one α-substituent is methyl and the other α-substituent is a second hydrocarbon linker, and ４ 222. The polypeptide of any one of claims 199-221, wherein one alpha substituent is methyl and the other alpha substituent is the second hydrocarbon linker.

223. the second hydrocarbon linker has the formula: 【Chemistry 45】 In the formula, one 【Chemistry 46】 is Xaa of the second hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 47】 is Xaa of the second hydrocarbon linker ４ 223. The polypeptide of claim 222, wherein the point of attachment to the alpha carbon atom is:

224. 224. The polypeptide of claim 223, wherein the second hydrocarbon crosslinker has the formula: 【Chemistry 48】

225. A polypeptide comprising the amino acid sequence SEQ ID NO:37, or a pharmaceutically acceptable salt thereof: 【Table 11】 During the ceremony, Xaa １５ and Xaa １０ are each independently an α,α-disubstituted amino acid; the polypeptide comprises at least two Nle; The polypeptide has a length of 7 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

226. 226. The polypeptide of claim 225, wherein the polypeptide comprises the amino acid sequence SEQ ID NO:37 plus at least one amino acid selected from L, E, R, H, S, Q, I, CBA, N-methyl Q, N-methyl E, N-methyl R, N-methyl D, N-methyl T, N-methyl I, Cpa, Cha, N-MeHis, N-MeCys, HomoHis, NHis, HomoR, Cit, Nar, Phe(4-guanidino), NMeGln, Nle, 2-Abu, Phe(4-Cl), 3,4-diClPh, 4-FPh, NptGly, NMeCha, Dcha, α-methyl L, allylGly, Alg, AC4C, A6C, Aze, (β-tBu-Ala), Tle, peptoid Q, DThr, and NMeLeu.

227. 227. The polypeptide of claim 226, wherein the polypeptide comprises the amino acid sequence SEQ ID NO:37 plus at least one amino acid selected from H, R, E, S, L, Q, I, CBA, and (2-Nal).

228. The polypeptide of any one of claims 225 to 227, wherein the polypeptide comprises an IQR.

229. 229. The polypeptide of claim 228, wherein the polypeptide comprises (Nle)IQR(Nle).

230. The polypeptide of any one of claims 225 to 229, wherein the polypeptide comprises an HRE and an LR.

231. 231. The polypeptide of any one of claims 225-230, wherein the polypeptide comprises L(2-Nal).

232. 231. The polypeptide of any one of claims 225 to 230, wherein the polypeptide comprises (CBA)(2-Nal).

233. the polypeptide HREXaa １５ SLQXaa １０ LR(Nle)IQR(Nle)L(2-Nal) (SEQ ID NO: 80), and HREXaa １５ SLQXaa １０ 226. The polypeptide of claim 225, wherein the polypeptide is selected from LR(Nle)IQR(Nle)(CBA)(2-Nal) (SEQ ID NO: 81).

234. 234. The polypeptide of any one of claims 225 to 233, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

235. Xaa １５ and Xaa １０ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

236. Xaa １５ and Xaa １０ is each (S)-2-(4'-pentenyl)alanine.

237. The polypeptide of any one of claims 225 to 233, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

238. The hydrocarbon linker has the formula: 【Chemistry 49】 In the formula, one 【Chemistry 50】 is Xaa of the hydrocarbon linker １５ indicates the point of attachment to the α carbon atom of 【Chemistry 51】 is Xaa of the hydrocarbon linker １０ 238. The polypeptide of claim 237, wherein the point of attachment to the alpha carbon atom is:

239. 198. The polypeptide of claim 197, wherein the hydrocarbon crosslinker has the formula: 【Chemistry 52】

240. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: 【Table 12】 During the ceremony, Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh; Xaa １１ is selected from R, N-methylR, E, K, homoR, Nar, and Cit, Xaa １１ and Xaa ５ at least one of contains E or K; A polypeptide, or a pharmaceutically acceptable salt thereof, wherein the polypeptide has a length of 8 to 30 amino acids.

241. Xaa １１ is E, and Xaa ５ The polypeptide of claim 240, wherein is R.

242. Xaa １１ is R, and Xaa ５ The polypeptide of claim 240, wherein is E.

243. Xaa １１ is K, and Xaa ５ The polypeptide of claim 240, wherein is R.

244. Xaa １１ is R, and Xaa ５ The polypeptide of claim 240, wherein is K.

245. 245. The polypeptide of any one of claims 240 to 244, wherein the polypeptide comprises L(2-Nal).

246. The polypeptide of any one of claims 240 to 245, wherein the polypeptide comprises IQR, IQE, or IQK.

247. The polypeptide of any one of claims 240 to 246, wherein the polypeptide comprises LQTLE, LQTLR, or LQRLK.

248. The polypeptide is selected from: LQTLEXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 98), LQTLRXaa ３ IQEXaa ６ L(2-Nal) (SEQ ID NO: 99), LQTLKXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 100), and LQTLRXaa ３ IQKXaa ６ L(2-Nal) (SEQ ID NO: 101), the polypeptide of claim 240.

249. 249. The polypeptide of any one of claims 240 to 248, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

250. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

251. Xaa ３ and Xaa ６ is each (S)-2-(4'-pentenyl)alanine.

252. The polypeptide of any one of claims 240 to 248, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

253. The hydrocarbon linker has the formula: 【Chemistry 53】 In the formula, one 【Chemical 54】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemistry 55】 is Xaa of the hydrocarbon linker ６ 253. The polypeptide of claim 252, wherein the point of attachment to the alpha carbon atom is:

254. 254. The polypeptide of claim 253, wherein the hydrocarbon crosslinker has the formula: 【Chemical Formula 56】

255. A polypeptide comprising the following amino acid sequence, or a pharmaceutically acceptable salt thereof: 【Table 13】 During the ceremony, Xaa ３ and Xaa ６ are each independently an α,α-disubstituted amino acid; Xaa ４ is selected from I, A, Nle, N-methyl I, CBA, and (DI); Xaa ８ is selected from Q and N-methyl Q; Xaa ５ is selected from R, A, Q, E, K, H, N-methylR, homoR, NMeArg, Nar, and Cit; Xaa ７ is selected from L, A, CBA, Cha, Cpa, Phe(4-Cl), (D-L), α-methyl L, DCcha, N-methyl Cha, allyl Gly, AC4C, A6C, Aze, NMeCha, (β-tBu-Ala), Tle, 4-FPh, and 3,4-diClPh, wherein The polypeptide comprises at least one of (DI) or (DL), The polypeptide has a length of 7 to 30 amino acids, or a pharmaceutically acceptable salt thereof.

256. Xaa ４ The polypeptide of claim 255, wherein is (DI).

257. Xaa ７ The polypeptide of claim 255, wherein is (DL).

258. 258. The polypeptide of any one of claims 255 to 257, wherein the polypeptide comprises a (DL)QTIR.

259. The polypeptide of any one of claims 255 to 257, wherein the polypeptide comprises LQT(DL)R.

260. The polypeptide is selected from: (DL)QTIRXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 102), LQT(DL)RXaa ３ IQRXaa ６ L(2-Nal) (SEQ ID NO: 103), LQTLRXaa ３ (DI)QRXaa ６ L(2-Nal) (SEQ ID NO: 104), and LQTLRXaa ３ IQRXaa ６ (DL)(2-Nal) (SEQ ID NO: 105), the polypeptide of claim 255.

261. 261. The polypeptide of any one of claims 255 to 260, wherein the α,α-disubstituted amino acid is an α-methyl, α-alkenyl amino acid.

262. Xaa ３ and Xaa ６ are each independently selected from (S)-2-(4'-pentenyl)alanine, (R)-2-(4'-pentenyl)alanine, (S)-2-(7'-octenyl)alanine, and (R)-2-(7'-octenyl)alanine.

263. Xaa ３ and Xaa ６ is each (S)-2-(4'-pentenyl)alanine.

264. 261. The polypeptide of any one of claims 255 to 260, wherein one α-substituent in the α,α-disubstituted amino acid is methyl and the other α-substituent in the α,α-disubstituted amino acid is a hydrocarbon linker.

265. The hydrocarbon linker has the formula: 【Chemical Formula 57】 In the formula, one 【Chemistry 58】 is Xaa of the hydrocarbon linker ３ indicates the point of attachment to the α carbon atom of 【Chemical Formula 59】 is Xaa of the hydrocarbon linker ６ 265. The polypeptide of claim 264, wherein the point of attachment to the alpha carbon atom is:

266. 266. The polypeptide of claim 265, wherein the hydrocarbon crosslinker has the formula: 【Chemistry 60】

267. 1. A polypeptide having a length of 6 to 30 amino acids, or a pharmaceutically acceptable salt thereof, wherein the polypeptide has at least 60% homology to a corresponding fragment of the wild-type HD2 domain of human B-cell CLL / lymphoma 9 (BCL9), and comprises at least one α-monosubstituted unnatural amino acid.

268. The polypeptide of claim 267, wherein the fragment of the wild-type HD2 domain of human B-cell CLL / lymphoma 9 (BCL9) is any fragment between positions 355 and 377 within BCL9.

269. 269. The polypeptide of claim 267 or claim 268, wherein the α-monosubstituted unnatural amino acid is selected from Nle, β-Ala, 2-Nal, β-L, and CBA.

270. 270. The polypeptide of claim 269, wherein the polypeptide comprises at least one 2-Nal.

271. 271. The polypeptide of claim 270, wherein the polypeptide comprises at least one CBA.

272. 272. The polypeptide of claim 270 or claim 271, wherein the polypeptide comprises at least one Nle.

273. 273. The polypeptide of claim 272, wherein the polypeptide comprises two Nles.

274. The polypeptide of any one of claims 267 to 273, wherein the polypeptide comprises at least one β-L.

275. 275. The polypeptide of any one of claims 267-274, wherein the polypeptide comprises at least one β-Ala.

276. 276. The polypeptide of claim 275, wherein said polypeptide comprises two β-Alas.

277. The polypeptide comprises at least one amino acid sequence selected from the following: 【Table 14】 A polypeptide according to any one of claims 267 to 276.

278. 278. The polypeptide of any one of claims 267 to 277, wherein the polypeptide comprises the amino acid sequence RSL.

279. 279. The polypeptide of any one of claims 267-278, wherein the polypeptide comprises an amino acid sequence selected from HRE and HQE.

280. 280. The polypeptide of any one of claims 267 to 279, wherein the polypeptide comprises the amino acid sequence QLE.

281. The polypeptide is selected from any one of the following polypeptides: 【Table 15】 The polypeptide described in claim 267.

282. 282. A pharmaceutically acceptable composition comprising the polypeptide of any one of claims 1 to 281, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

283. 1. A method comprising: a method of inhibiting the binding of BCL9 to β-catenin in a subject, and / or a method of inhibiting canonical Wnt / β-catenin signaling in a subject, and / or a method for decreasing the survival of regulatory T cells in a subject, and / or a method for reducing the expression of VEGF in a tumor in a subject, and / or a method for increasing the infiltration of CD4+ T cells and CD8+ T cells into a tumor in a subject; and / or a method for increasing T helper 17 (Th17) cells to tumors in a subject; and / or a method for reducing intratumoral dendritic cells in a subject; and / or A half-life (T) of at least 2 hours when administered to a subject １／２ ) and / or - a method for inducing a tumor microenvironment favorable to an immune response in a subject; and / or a method of inhibiting tumor growth in a subject, and / or a method for inhibiting cancer stem cell proliferation in a subject, and / or a method for inhibiting tumor metastasis in a subject, and / or • a method of treating cancer in a subject; 282. A method comprising administering to a subject in need thereof a therapeutically effective amount of the polypeptide of any one of claims 1 to 281 or the pharmaceutical composition of claim 282.

284. 284. The method of claim 283, wherein the cancer is familial adenomatous polyposis (FAP), eye cancer, rectal cancer, colon cancer, colorectal cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, benign and malignant tumors, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, kidney cancer, brain / CNS cancer, throat cancer, multiple myeloma, cutaneous melanoma, acute lymphocytic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, oral / pharyngeal cancer, esophageal cancer, laryngeal cancer, lymphoma, neurofibromatosis, tuberous sclerosis, hemangioma, gastric cancer, ovarian cancer, hepatocellular carcinoma, or lymphangiogenesis.

285. 285. The method of claim 284, wherein the cancer is colorectal cancer.

286. The method of claim 284, wherein the cancer is gastric cancer.

287. 285. The method of claim 284, wherein the cancer is ovarian cancer.

288. The method of claim 284, wherein the cancer is hepatocellular carcinoma.

289. 285. The method of claim 284, wherein the cancer is breast cancer.

290. 285. The method of claim 284, wherein the cancer is prostate cancer.

291. 285. The method of claim 284, wherein the cancer is cutaneous melanoma.

292. 285. The method of claim 284, wherein the cancer is lung cancer.

293. 285. The method of claim 284, further comprising administering at least one additional agent.

294. 294. The method of claim 293, wherein the at least one additional agent is selected from the group consisting of a checkpoint inhibitor, an EGFR inhibitor, a VEGF inhibitor, a chemotherapeutic agent, and a VEGFR inhibitor.

295. 295. The method of claim 294, wherein the checkpoint inhibitor is an anti-PD-1 antibody, an anti-PD-L1 antibody, or an anti-CTLA4 antibody.

296. 295. The method of claim 294, wherein said checkpoint inhibitor targets a stimulatory checkpoint molecule selected from the group consisting of CD27, CD40, OX40, GITR, and CD137.

297. 295. The method of claim 294, wherein said checkpoint inhibitor targets an inhibitory checkpoint molecule selected from the group consisting of A2AR, B7-H3, B7-H4, attenuator of B and T lymphocytes (BTLA), indoleamine 2,3-dioxygenase (IDO), killer cell immunoglobulin-like receptor (KIR), lymphocyte activation gene 3 (LAG3), T-cell immunoglobulin and mucin domain 3 (TIM-3), VISTA (C10orf54), and V domain Ig suppressor of T-cell activation.

298. 295. The method of claim 294, wherein the EGFR inhibitor is erlotinib, gefitinib, lapatinib, panitumumab, vandetanib, or cetuximab.

299. 295. The method of claim 294, wherein the VEGF inhibitor or VEGFR inhibitor is pazopanib, bevacizumab, sorafenib, sunitinib, axitinib, ponatinib, regorafenib, vandetanib, cabozantinib, ramucirumab, lenvatinib, or ziv-aflibercept.

300. The chemotherapeutic agent may be cyclophosphamide, methotrexate, 5-fluorouracil (5-FU), doxorubicin, mustine, vincristine, procarbazine, prednisolone, dacarbazine, bleomycin, etoposide, cisplatin, epirubicin, capecitabine, folinic acid, actinomycin, all-trans retinoic acid, azacitidine, azathioprine, bortezomib, carboplatin, chlorambucil, 295. The method of claim 294, wherein the compound is cytarabine, daunorubicin, docetaxel, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, irinotecan, mechlorethamine, mercaptopurine, mitoxantrone, paclitaxel, pemetrexed, teniposide, thioguanine, topotecan, valrubicin, vinblastine, vindesine, vinorelbine, or oxaliplatin.

301. 284. The method of claim 283, further comprising exposing the subject to radiation therapy and / or chemotherapy.

302. The method of claim 283, further comprising measuring at least one biomarker to monitor treatment / inhibition effectiveness and / or to select subjects for treatment.

303. 303. The method of claim 302, wherein the biomarker is one or more of BCL9, CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin.

304. 303. The method of claim 302, wherein a decrease in gene expression and / or protein levels of CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin indicates therapeutic / inhibitory efficacy and / or the subject is selected for treatment if gene expression and / or protein levels of CD44, Axin2, cMyc, LGR5, VEGFA, Sox2, Oct4, Nanog, and / or active beta-catenin are elevated.

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