Stapled peptides and methods thereof

Abstract

In particular, the present disclosure provides various useful agents. In some embodiments, the agents provided can bind to beta-catenin. In some embodiments, the present disclosure provides techniques for modulating beta-catenin function. In some embodiments, the present disclosure provides techniques for preventing and / or treating conditions, disorders, or diseases associated with beta-catenin. In some embodiments, the present disclosure provides designed amino acids and agents that can provide improved properties and / or activities.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application Nos. 63 / 208,487, filed June 8, 2021, 63 / 224,834, filed July 22, 2021, and 63 / 303,952, filed January 27, 2022, each of which is incorporated by reference herein in its entirety. [Background technology]

[0002] background Stapled peptides are useful in a variety of applications, for example, as biologically active agents, they can be utilized to modulate various biological functions. Summary of the Invention [Means for solving the problem]

[0003] overview Among other things, the present disclosure provides powerful techniques (e.g., agents (e.g., peptides, which in many embodiments are or include stapled peptides), compositions, methods, etc.) for modulating various biological functions.

[0004] In some embodiments, the present disclosure provides agents, e.g., stapled peptides, comprising multiple staples. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staples. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staples within 10-20 amino acid residues, e.g., 10-15, 11-15, 11-14, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive amino acid residues. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staples within 11 consecutive amino acid residues. In some embodiments, the present disclosure provides agents, e.g., stapled peptides comprising three or more staples within 14 consecutive amino acid residues. In some embodiments, there are three staples within such a number of amino acid residues. In some embodiments, there are four staples within such number of consecutive amino acid residues. Without intending to be limited by theory, in some embodiments, provided agents, e.g., stapled peptides, have increased rigidity relative to a reference peptide (e.g., a non-stapled peptide, or a stapled peptide having fewer staples (in some embodiments, fewer staples within a certain number of amino acid residues as described herein), etc.). In some embodiments, provided agents, e.g., stapled peptides, exhibit various desired properties and / or activities. In some embodiments, provided agents, e.g., stapled peptides, provide improved desired properties and / or activities relative to a reference peptide (e.g., a non-stapled peptide, or a stapled peptide having fewer staples (in some embodiments, fewer staples within a certain number of amino acid residues as described herein), etc.).

[0005] In some embodiments, the provided technology includes designed structural features, e.g., novel amino acid residues, which can provide significantly improved properties and / or activity compared to comparable reference technology that does not contain such designed structural features. In some embodiments, the present disclosure provides designed amino acids as described herein, which, when incorporated into peptide agents, e.g., stapled peptides, can provide significantly improved properties and / or activity, e.g., improved lipophilicity and / or cellular delivery, compared to reference amino acids (e.g., Asp). In some embodiments, the present disclosure provides technology including peptides containing such designed amino acid residues. In some embodiments, the present disclosure provides stapled peptides containing such designed amino acid residues.

[0006] In some embodiments, the present disclosure provides techniques for modulating one or more functions of beta-catenin. Specifically, in some embodiments, the present disclosure provides various agents, such as peptides, in many cases stapled peptides, that can bind to beta-catenin and modulate its function. As shown herein, in some embodiments, the present disclosure binds to agents that can interact with beta-catenin at a unique set of residues. In some embodiments, the binding site includes one or more or all of the set of residues. In some embodiments, provided agents interact with one or more of a set of residues that are or correspond to the following residues of SEQ ID NO:1: A305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419. In some embodiments, provided agents interact with one or more of the amino acid residues that are or correspond to A305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419 of SEQ ID NO:1. In some embodiments, provided agents interact with one or more of the amino acid residues that are or correspond to A305, Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419 of SEQ ID NO: 1. In some embodiments, provided agents interact with one or more of the amino acid residues that are or correspond to G307, K312, K345, W383, N387, D413, and N415 of SEQ ID NO: 1. In some embodiments, provided agents interact with one or more of the amino acid residues that are or correspond to K312, K345, R386, and W383 of SEQ ID NO: 1.In some embodiments, provided agents interact with one or more of the set of residues that are or correspond to the following residues in SEQ ID NO: 1: G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, provided agents interact with all of the set of residues that are or correspond to the following residues in SEQ ID NO: 1: Y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, provided agents interact with all of the set of residues that are or correspond to the following residues in SEQ ID NO: 1: G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, provided agents interact with all of a set of residues that are or correspond to the following residues of SEQ ID NO: 1: Y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416. In some embodiments, provided agents interact with one or more of the amino acid residues that are or correspond to K312, K345, and W383 of SEQ ID NO: 1. In some embodiments, provided agents interact with amino acid residues that are or correspond to K312, K345, and W383 of SEQ ID NO: 1.

[0007] As demonstrated herein, the provided technology can modulate one or more biological processes associated with beta-catenin. In some embodiments, provided agents, e.g., stapled peptides, compete with a ligand (e.g., a member of the T-cell factor / lymphoid enhancer factor (TCF / LEF) family of transcription factors) for binding to beta-catenin. In some embodiments, provided agents compete with a ligand (e.g., a member of the T-cell factor / lymphoid enhancer factor (TCF / LEF) family of transcription factors at the TCF site on beta-catenin) for binding to a specific binding site on beta-catenin. In some embodiments, provided technology competes with TCF for interaction with beta-catenin. In some embodiments, binding of a provided agent to the beta-catenin site reduces, inhibits, and / or blocks binding of another binding partner (e.g., a kinase) to beta-catenin. In some embodiments, binding of a provided agent blocks binding of beta-catenin by a TCF / LEF family member. In some embodiments, the present disclosure provides agents that can selectively bind to a site on beta-catenin by other ligands (e.g., peptides, proteins, etc.; in some embodiments, the ligand is axin, and in some embodiments, the ligand is Bcl9) over one of many other binding sites. In some embodiments, the provided technology modulates one or more beta-catenin functions associated with beta-catenin's interaction with TCFs. In some embodiments, the provided technology selectively modulates beta-catenin functions, e.g., functions associated with TCF interaction. In some embodiments, the provided technology selectively modulates beta-catenin function and does not significantly affect functions not associated with beta-catenin (e.g., various functions and / or processes in the Wnt pathway not associated with beta-catenin). In some embodiments, the provided technology is useful for inhibiting beta-catenin function. In some embodiments, the provided technology is useful for promoting and / or enhancing immune activity, e.g., anti-tumor adaptive immunity.

[0008] In some embodiments, the provided technology is useful for preventing or treating various conditions, disorders, or diseases, including cancer. In some embodiments, the present disclosure provides a method for treating or preventing a condition, disorder, or disease associated with beta-catenin, comprising administering to a subject suffering from or susceptible thereto an effective amount of a provided agent, or a pharmaceutically acceptable salt thereof. In some embodiments, the condition, disorder, or disease is associated with the interaction of beta-catenin with TCF. In some embodiments, the agent, e.g., a stapled peptide, is administered as a pharmaceutical composition. In some embodiments, the present disclosure provides a pharmaceutical composition comprising or delivering a provided agent, or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition further comprises a lipid. As provided herein, in some embodiments, a suitable lipid can facilitate delivery / activity. In some embodiments, the agent is or comprises a peptide. In some embodiments, the agent is or comprises a stapled peptide. In some embodiments, the provided agent capable of binding to beta-catenin comprises one or more designed amino acid residues.

[0009] In some embodiments, the disclosure provides an agent that binds to a polypeptide comprising or consisting of SEQ ID NO:1 (Uniprot identifier P35222), or to residues 250-450 of SEQ ID NO:1, or to residues 305-419 of SEQ ID NO:1. Uniprot number P35222 MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPSLSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLDEGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDEDQVVVNKAAVMVHQLSKKEASRHAIMRSPQMV SAIVRTMQNTNDVETARCTAGTLHNLSHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKMVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTSRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSD AATKQEGMEGLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDREDITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNLALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGALHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEGATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETADLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGLPDLGHAQDLMDGLPPGDSNQLAWFDTDL (SEQ ID NO: 1)

[0010] In some embodiments, provided agents specifically interact with one or more residues that are or correspond to residues 305-419 of SEQ ID NO: 1. In some embodiments, provided agents specifically bind to a motif (e.g., a portion of a polypeptide, a domain of a polypeptide, etc.) that includes one or more residues that correspond to Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Arg342, Lys345, Val346, Val349, Gln375, Arg376, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419 of SEQ ID NO: 1. In some embodiments, the provided agents specifically bind to a motif (e.g., a portion of a polypeptide, a domain of a polypeptide, etc.) that includes one or more residues corresponding to Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Lys345, Val346, Val349, Gln375, Arg376, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419 of SEQ ID NO:1. In some embodiments, the agent binds to a motif that includes one or more of the following residues in SEQ ID NO:1: Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Arg342, Lys345, Val346, Val349, Gln375, Arg376, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419. In some embodiments, the agent specifically binds to a motif that includes one or more of the following residues in SEQ ID NO:1: Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Lys345, Val346, Val349, Gln375, Arg376, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419.In some embodiments, the agent binds to a motif that includes one or more of the following residues in SEQ ID NO:1: Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Arg342, Lys345, Val346, Val349, Gln375, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419. In some embodiments, the agent binds to a motif that includes one or more of the following residues in SEQ ID NO: 1: Ala305, Tyr306, Gly307, Asn308, Gln309, Lys312, Lys345, Val346, Val349, Gln379, Asn380, Leu382, Trp383, Arg386, Asn387, Asp413, Asn415, Val416, Thr418, and Cys419. In some embodiments, the provided technology binds to a motif that includes at least two, three, four, five, or six of G307, K312, K345, W383, N387, and N415. In some embodiments, provided techniques bind to motifs including at least two, three, four, five, six, or seven of G307, K312, K345, W383, N387, D413, and N415. In some embodiments, provided agents specifically bind to such motifs. In some embodiments, motifs may be referred to as binding sites. In some embodiments, provided techniques selectively bind to such binding sites over axin binding sites. In some embodiments, provided techniques selectively bind to such binding sites over Bcl9 binding sites. In some embodiments, provided techniques selectively bind to such binding sites over TCF binding sites. In some embodiments, provided techniques bind to such binding sites in a reverse N to C orientation compared to TCF. In some embodiments, provided techniques do not bind to the axin binding site of beta-catenin. In some embodiments, provided techniques do not bind to the Bcl9 binding site of beta-catenin. In some embodiments, provided techniques do not bind to the ICAT binding site of beta-catenin.A variety of techniques, such as crystallography, NMR, biochemical assays, etc., may be used to assess interactions with beta-catenin in accordance with the present disclosure.

[0011] In some embodiments, the provided technology provides agents, e.g., stapled peptides, comprising three staples of no more than 10-20, 10-15, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 10 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 11 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 12 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 13 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 14 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 15 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 16 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 17 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 18 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 19 consecutive amino acid residues. In some embodiments, there are three or more staples of no more than 20 consecutive amino acid residues. In some embodiments, two staples are attached to the same amino acid residue. In some embodiments, two staples are attached to the same backbone atom. In some embodiments, two staples are attached to the same backbone carbon atom. In some embodiments, two staples are attached to the alpha-carbon atom of an amino acid residue and each independently attaches to a different amino acid residue.

[0012] In some embodiments, the first staple in the agent, e.g., stapled peptide, is attached to amino acid residues at positions i and i+3. In some embodiments, there is a second staple attached to amino acid residues at positions i+3 and i+10. In some embodiments, there is a third staple attached to amino acid residues at positions i+9 and i+13. As used in the art, those skilled in the art will understand that i, i+3, i+9, i+10, i+13, etc. are routinely used to denote relative positions of amino acid residues. In some embodiments, they may also denote absolute positions in the agent, e.g., peptide. In some embodiments, i is an integer from 1 to 50 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). In some embodiments, i is 1. In some embodiments, there is a fourth staple in the agent, e.g., stapled peptide.

[0013] In some embodiments, there are two amino acid residues between two amino acid residues attached to the same staple. Such a staple may be referred to as an (i, i+3) staple. Similarly, in some embodiments, there are three, four, five, six, seven, eight, nine, or ten amino acid residues between two amino acid residues attached to the same staple. Such staples may be referred to as (i, i+4), (i, i+5), (i, i+6), (i, i+7), (i, i+8), (i, i+9), (i, i+10), or (i, i+11) staples, respectively.

[0014] In some embodiments, the agent, e.g., the stapled peptide, comprises an (i, i+2) staple and an (i, i+7) staple. In some embodiments, the agent, e.g., the stapled peptide, comprises an (i, i+3) staple and an (i, i+7) staple. In some embodiments, the (i, i+3) staple and the (i, i+7) staple are attached to the same amino acid residue. In some embodiments, the (i, i+3) staple and the (i, i+7) staple are attached to the same atom. In some embodiments, the (i, i+3) staple and the (i, i+7) staple are attached to the same alpha carbon atom. For example, in compound I-1, the (i, i+3) staple is attached to the amino acid residues at positions 1 and 4, and the (i, i+7) staple is attached to the amino acid residues at positions 4 and 11, and both staples are attached to the alpha carbon of the amino acid residue at position 4. In some embodiments, the agent further comprises a third staple. In some embodiments, the third staple is (i, i+4). In some embodiments, the third staple is (i, i+7). In some embodiments, the third staple is not bound to any of the amino acid residues bound to the first two staples. In some embodiments, the agent further comprises a fourth staple. In some embodiments, the fourth staple is (i, i+4). In some embodiments, the fourth staple is (i, i+7). In some embodiments, the fourth staple is not bound to any of the amino acid residues bound to the first two staples. In some embodiments, the fourth staple is not bound to any of the amino acid residues bound to the first or third staples.

[0015] In some embodiments, a provided agent, e.g., a peptide agent such as a stapled peptide agent, comprises one or more (e.g., 1, 2, 3, 4, 5, 6, or 7) of the following group (in some embodiments, in the N to C direction): a first acidic group (e.g., of a first acidic amino acid residue), a second acidic group (e.g., of a second acidic amino acid residue), optionally a third acidic group (e.g., of a third acidic amino acid residue); Optionally, hydrophobic groups (e.g., those of hydrophobic amino acid residues) a first aromatic group (e.g., of a first aromatic amino acid residue); a second aromatic group (e.g., of the first aromatic amino acid residue), and a third aromatic group (eg, that of a third aromatic amino acid residue). In some embodiments, the agent comprises a first and a second acidic group and a first, a second, and a third aromatic group. In some embodiments, such an agent additionally comprises a third acidic group (e.g., of a third acidic amino acid residue) and / or a hydrophobic group (e.g., of a hydrophobic amino acid residue). In some embodiments, such an agent additionally comprises a third acidic group (e.g., of a third acidic amino acid residue) and a hydrophobic group (e.g., of a hydrophobic amino acid residue).In some embodiments, the distance between the first acidic group and the second acidic group is approximately the distance between the acidic groups of two acidic amino acid residues of the peptide motif, where there are two amino acid residues between the two acidic amino acid residues (e.g., if the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position N+3); the distance between the first acidic group and the third acidic group (if present) is approximately the distance between the acidic groups of two acidic amino acid residues of the peptide motif, where there are three amino acid residues between the two acidic amino acid residues (e.g., if the first acidic amino acid residue is at position N, the third acidic amino acid residue is at position N+4); the distance between the first acidic group and the hydrophobic group (if present) is approximately the distance between the acidic group of the acidic amino acid residue and the hydrophobic group of the hydrophobic amino acid residue of the peptide motif, where there are five amino acid residues between the first acidic amino acid residue and the hydrophobic amino acid residue (e.g., if the first acidic amino acid residue is at position N, the hydrophobic amino acid residue is at position N+6); and the first aromatic group is the approximate distance between the acidic group of the first acidic amino acid residue and the aromatic group of the aromatic amino acid residue of the peptide motif, where there are six amino acid residues between the first acidic amino acid residue and the first aromatic amino acid residue (e.g., if the first acidic amino acid residue is at position N, the first aromatic amino acid residue is at position N+7); the distance between the first aromatic group and the second aromatic group is the approximate distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, where there are two amino acid residues between the two aromatic amino acid residues (e.g., if the first aromatic amino acid residue is at position M, the second aromatic amino acid residue is at position M+3); and / or the distance between the first aromatic group and the third aromatic group is the approximate distance between the aromatic groups of two aromatic amino acid residues of the peptide motif, where there are three amino acid residues between the two aromatic amino acid residues (e.g., if the first aromatic amino acid residue is at position M, the third aromatic amino acid residue is at position M+4). In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position N+3, and the first, second, and third aromatic amino acid residues are at positions N+7, N+10, and N+11, respectively.In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position N+3, the third acidic amino acid residue is at position N+4, and the first, second, and third aromatic amino acid residues are at positions N+7, N+10, and N+11, respectively. In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position N+3, the hydrophobic amino acid residue is at position N+6, and the first, second, and third aromatic amino acid residues are at positions N+7, N+10, and N+11, respectively. In some embodiments, the first acidic amino acid residue is at position N, the second acidic amino acid residue is at position N+3, the third acidic amino acid residue is at position N+4, the hydrophobic amino acid residue is at position N+6, and the first, second, and third aromatic amino acid residues are at positions N+7, N+10, and N+11, respectively. In some embodiments, M is N+7. In some embodiments, N is 1-7. In some embodiments, N is 1, 2, 3, 4, or 5. In some embodiments, N is 1. In some embodiments, N is 2. In some embodiments, N is 3. In some embodiments, N is 4. In some embodiments, N is 5. In some embodiments, M is 8-16. In some embodiments, M is 8. In some embodiments, M is 9. In some embodiments, M is 10. In some embodiments, M is 11. In some embodiments, M is 12. In some embodiments, M is 13. In some embodiments, the peptide motif is an alpha-helical motif, and each amino acid residue is independently an alpha amino acid residue. In some embodiments, the peptide motif is stapled. In some embodiments, there are two or more staples in the peptide motif; in some embodiments, there are three staples; in some embodiments, there are four staples; in some embodiments, there are four or more staples.In some embodiments, the peptide motif is or includes an agent described in the Tables herein (e.g., I-xxxx, where xxxx is a number (e.g., I-1, I-10, I-100, I-1000, etc.)). In some embodiments, the first acidic group is X as described herein. 2 and the second acidic group is X as described herein. 5 and the third acidic group (if present) is selected from the group consisting of X 6 and the hydrophobic group (if present) is X as defined herein. 8 wherein the first aromatic group is X as described herein. 9 and the second aromatic group is X as described herein. 12 and / or the third aromatic group is X as described herein. 13In some embodiments, as described herein, provided agents are staple peptides that contain one or more staples. In some embodiments, as described herein, provided agents are staple peptides that contain two or more staples. In some embodiments, as described herein, provided agents are staple peptides that contain three or more staples. In some embodiments, when contacting a beta-catenin polypeptide, the first acidic group interacts with Lys312 and / or Gly307, or the amino acid residues corresponding thereto, the second acidic group interacts with Asn387, Trp383, and / or Arg386, or the amino acid residues corresponding thereto, the first aromatic group interacts with Lys345 and / or Trp383, or the amino acid residues corresponding thereto, the second aromatic group interacts with Trp383 and / or Asn415, or the amino acid residues corresponding thereto, and the third aromatic group interacts with Gln379, Leu383, Val416, Asn415, and / or Trp383, or the amino acid residues corresponding thereto. In some embodiments, the third acidic group interacts with Asn387, Trp383, and / or Arg386, or the amino acid residues corresponding thereto. In some embodiments, the hydrophobic group interacts with Trp383 or a corresponding amino acid residue.

[0016] In some embodiments, the present disclosure provides a compound of formula I R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C I or a salt thereof, wherein each variable is independently as described herein.

[0017] In some embodiments, the present disclosure provides: [X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 or a drug comprising the same, During the ceremony, each of p0, p15, p16, and p17 is independently 0 or 1; X 0 , X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , and X 17 are independently an amino acid residue, Provide medication.

[0018] In some embodiments, the present disclosure provides: [X] p X 1 X 2 X 3 X4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X] p’ or a drug comprising the same, During the ceremony, each of p15, p16, and p17 is independently 0 or 1; each of p and p' is independently 0 to 10; X, X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , and X 17 are independently an amino acid residue, Provide medication.

[0019] In some embodiments, the drug is R N -[X]pX 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 [X 14 ] p14 [X15 ] p15 [X 16 ] p16 [X 17 ] p17 [X]p'-R C wherein each variable is independently as described herein.

[0020] In some embodiments, the agent is X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 [X 14 ] p14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 [X 18 ] p18 [X 19 ] p19 [X 20 ] p20 [X 21 ] p21 [X 22 ] p22 [X 23 ] p23 wherein each of p14, p15, p16, p17, p18, p19, p20, p21, p22, and p23 is independently 0 or 1; and X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17, X 18 , X 19 , X 20 , X 21 , X 22 , and X 23 are each independently an amino acid residue described herein.

[0021] In some embodiments, such peptides contain three or more staples, hi some embodiments, such peptides contain five or more residues suitable for stapling.

[0022] In some embodiments, the present disclosure provides a method for producing a compound of formula [X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 or a drug comprising the same, During the ceremony, each of p0, p15, p16, and p17 is independently 0 or 1; X 0 , X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , and X17 are independently an amino acid residue; X 2 contains a side chain containing an acidic or polar group; X 5 contains a side chain containing an acidic or polar group; X 13 contains a side chain containing an optionally substituted aromatic group; X 1 , X 3 , X 4 , X 7 , X 10 , X 11 , and X 14 are each independently an amino acid residue suitable for stapling or are each independently stapled, Provide medication.

[0023] In some embodiments, the present disclosure provides: [X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X 14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 or a drug comprising the same, During the ceremony, each of p0, p15, p16, and p17 is independently 0 or 1; X 0 , X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , and X 17 are independently an amino acid residue; X 2 contains a side chain containing an acidic or polar group; X 5 contains a side chain containing an acidic or polar group; X 6 contains a side chain containing an acidic or polar group; X 13 contains a side chain containing an optionally substituted aromatic group; X 1 , X 3 , X 4 , X 7 , X 10 , X 11 , and X 14 are each independently an amino acid residue suitable for stapling or are each independently stapled, Provide medication.

[0024] In some embodiments, the agent is or comprises a peptide. In some embodiments, the agent is or comprises a stapled peptide. In some embodiments, the agent is a peptide. In some embodiments, the agent is a stapled peptide. In some embodiments, the agent, peptide, or stapled peptide is a peptide having a structure of [X 0 ] p0 X 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 X 10 X 11 X 12 X 13 X14 [X 15 ] p15 [X 16 ] p16 [X 17 ] p17 In some embodiments, X has the structure 1 and X 4 , and / or X 4 and X 11 are independently amino acid residues suitable for stapling or are stapled, or X 3 and X 10 are independently amino acid residues suitable for stapling or are stapled. 1 and X 4 are independently amino acid residues suitable for stapling. 1 and X 4 In some embodiments, X 4 and X 11 are independently amino acid residues suitable for stapling. 4 and X 11 In some embodiments, X 1 and X 4 , and X 4 and X 11 are independently amino acid residues suitable for stapling. In some embodiments, the staple peptide is a stitch peptide comprising two or more staples, some of which may be attached to the same backbone atom. In some embodiments, X 1 and X 4 The X is stapled. 4 and X 11 In some embodiments, X 1 and X 4 Staples to connect the X 4 and X 11 The staples connecting the X 4In some embodiments, the common backbone atom is X 4 In some embodiments, X is the alpha carbon of 3 and X 10 are independently amino acid residues suitable for stapling. 3 and X 10 In some embodiments, X 1 and X 3 are independently amino acid residues suitable for stapling. 1 and X 3 In some embodiments, X 10 and X 14 are independently amino acid residues suitable for stapling. 10 and X 14 In some embodiments, X 7 and X 10 are independently amino acid residues suitable for stapling. 7 and X 10 In some embodiments, X 7 and X 14 are independently amino acid residues suitable for stapling. 7 and X 14 In some embodiments, X 3 and X 7 are independently amino acid residues suitable for stapling. 3 and X 7 is stapled.

[0025] In some embodiments, the present disclosure provides an agent that binds to a polypeptide comprising or consisting of residues 305-419 of SEQ ID NO: 1 as described herein. In some embodiments, the agent, e.g., peptide, has a molecular mass of no more than about 5000 daltons. In some embodiments, it is no more than about 2500 daltons, no more than 3000 daltons, no more than 3500 daltons, no more than 4000 daltons, no more than 4500 daltons, or no more than 5000 daltons. In some embodiments, it is no more than about 2500 daltons. In some embodiments, it is no more than about 3000 daltons. In some embodiments, it is no more than about 3500 daltons. In some embodiments, it is no more than about 4000 daltons. In some embodiments, it is no more than about 500 daltons.

[0026] In some embodiments, the present disclosure provides various techniques, e.g., reagents, methods, etc., for preparing, characterizing, evaluating, and using the provided agents and compositions thereof. In some embodiments, the present disclosure provides methods, reagents, and / or systems, e.g., for identifying, characterizing, and / or evaluating the provided agents and their uses (e.g., as therapeutic or diagnostic agents).

[0027] In some embodiments, the present disclosure provides a pharmaceutical composition comprising or delivering a provided agent and a pharmaceutically acceptable carrier. In some embodiments, the provided agent is in a pharmaceutically acceptable salt form. In some embodiments, the provided composition comprises a pharmaceutically acceptable salt form of the agent. In some embodiments, in various compositions and methods, the agent is provided as a pharmaceutically acceptable salt form.

[0028] In some embodiments, the present disclosure provides a method of modulating the properties, activity, and / or function of beta-catenin, comprising contacting beta-catenin with a provided agent. In some embodiments, the present disclosure provides a method of modulating the properties, activity, and / or function of beta-catenin in a system containing beta-catenin, comprising administering to the system an effective amount of a provided agent. In some embodiments, the present disclosure provides a method of modulating the properties, activity, and / or function of beta-catenin in a system expressing beta-catenin, comprising administering or delivering to the system an effective amount of a provided agent. In some embodiments, the activity of beta-catenin is inhibited or reduced. In some embodiments, the function of beta-catenin is inhibited or reduced. In some embodiments, the property, activity, and / or function is associated with beta-catenin / TCF interaction.

[0029] In some embodiments, the present disclosure provides methods of modulating beta-catenin / TCF interaction. In some embodiments, the present disclosure provides methods of modulating beta-catenin / TCF interaction, comprising contacting beta-catenin with a provided agent. In some embodiments, the present disclosure provides methods of modulating beta-catenin / TCF interaction in a system comprising beta-catenin and TCF, comprising administering or delivering to the system an effective amount of a provided agent. In some embodiments, the present disclosure provides methods of modulating beta-catenin / TCF interaction in a system expressing beta-catenin and TCF, comprising administering or delivering to the system an effective amount of a provided agent. In some embodiments, the interaction between beta-catenin and TCF is reduced. In some embodiments, the interaction between beta-catenin and TCF is inhibited.

[0030] In some embodiments, the present disclosure provides a method of inhibiting cell proliferation, comprising administering or delivering an effective amount of a provided agent to a population of cells. In some embodiments, the present disclosure provides a method of inhibiting cell proliferation in a system, comprising administering or delivering an effective amount of a provided agent to the system. In some embodiments, the present disclosure provides a method of inhibiting cell growth, comprising administering or delivering an effective amount of a provided agent to a population of cells. In some embodiments, the present disclosure provides a method of inhibiting cell growth in a system, comprising administering or delivering an effective amount of a provided agent to the system. In some embodiments, such cell proliferation is beta-catenin dependent. In some embodiments, such cell growth is beta-catenin dependent. In some embodiments, such proliferation or growth is dependent on the interaction of beta-catenin with TCF.

[0031] In some embodiments, the present disclosure provides methods for reducing or preventing activation of the WNT pathway. In some embodiments, the present disclosure provides methods for reducing or preventing activation of the WNT pathway in a system, the method comprising administering or delivering to the system an effective amount of a provided agent.

[0032] In some embodiments, the system is in vitro. In some embodiments, the system is ex vivo. In some embodiments, the system is in vivo. In some embodiments, the system is or comprises a cell. In some embodiments, the system is or comprises a tissue. In some embodiments, the system is or comprises an organ. In some embodiments, the system is or comprises an organism. In some embodiments, the system is an animal. In some embodiments, the system is a human. In some embodiments, the system is or comprises a cell, tissue, or organ associated with a condition, disorder, or disease. In some embodiments, the system is or comprises a cancer cell.

[0033] In some embodiments, the present disclosure provides methods for preventing a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for preventing a condition, disorder, or disease, comprising administering or delivering to a subject susceptible thereto an effective amount of an agent of the present disclosure. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease, comprising administering or delivering to a subject susceptible thereto an effective amount of an agent of the present disclosure. In some embodiments, the present disclosure provides methods for reducing the risk of a condition, disorder, or disease in a population, comprising administering or delivering to a population of subjects susceptible thereto an effective amount of an agent of the present disclosure. In some embodiments, the present disclosure provides methods for treating a condition, disorder, or disease. In some embodiments, the present disclosure provides methods for treating a condition, disorder, or disease, comprising administering or delivering to a subject suffering therefrom an effective amount of an agent of the present disclosure. In some embodiments, symptoms are reduced, eliminated, or prevented. In some embodiments, one or more parameters for assessing the condition, disorder, or disease are improved. In some embodiments, the subject's survival time is extended. As will be appreciated by one of skill in the art, in some embodiments, the effects of prevention, risk reduction, and / or treatment can be evaluated through clinical trials and observed in a subject population. In some embodiments, the condition, disorder, or disease is cancer. In some embodiments, the condition, disorder, or disease is associated with beta-catenin. In some embodiments, the condition, disorder, or disease is associated with the interaction of beta-catenin with TCF. In some embodiments, the condition, disorder, or disease is bladder cancer. In some embodiments, the condition, disorder, or disease is endometrial cancer. In some embodiments, the condition, disorder, or disease is adrenocortical carcinoma. In some embodiments, the condition, disorder, or disease is gastric cancer. In some embodiments, the condition, disorder, or disease is lung cancer. In some embodiments, the condition, disorder, or disease is melanoma. In some embodiments, the condition, disorder, or disease is esophageal cancer.In some embodiments, the condition, disorder, or disease is colorectal cancer. In some embodiments, the cancer is liver cancer. In some embodiments, the cancer is prostate cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is endometrial cancer. Mutations that result in constitutive activation of Wnt / beta-catenin-mediated signaling have been reported to be present in approximately 20% of all human cancers. In some embodiments, the condition, disorder, or disease is associated with WNT signaling. In some embodiments, the condition, disorder, or disease is associated with beta-catenin-dependent WNT signaling. In some embodiments, the condition, disorder, or disease is associated with beta-catenin / TCF interaction. In some embodiments, it has been reported that beta-catenin / TCF interaction can promote cell proliferation, epithelial-mesenchymal transition (EMT), cancer stem cell phenotype, and the like.

[0034] In some embodiments, the agent is administered as a pharmaceutical composition that contains or delivers such an agent.In some embodiments, the agent is provided and / or delivered in a pharmaceutically acceptable salt form.In some embodiments, in the composition (for example, a liquid composition at a certain pH), the agent can be present in various forms, including various pharmaceutically acceptable salt forms.

[0035] In some embodiments, the provided agent is utilized in combination with a second therapy. In some embodiments, the provided agent is utilized in combination with a second therapeutic agent. In some embodiments, the second therapy or therapeutic agent is administered prior to administration or delivery of the provided agent. In some embodiments, the second therapy or therapeutic agent is administered approximately simultaneously with administration or delivery of the provided agent. In some embodiments, the second therapy or therapeutic agent is administered after administration or delivery of the provided agent. In some embodiments, the subject is exposed to both the provided agent and the second therapeutic agent. In some embodiments, the subject is exposed to the therapeutic effects of the provided agent and the second therapeutic agent. In some embodiments, the second therapy is or includes surgery. In some embodiments, the second therapy is or includes radiation therapy. In some embodiments, the second therapy is or includes immunotherapy. In some embodiments, the second therapeutic agent is or includes a drug. In some embodiments, the second therapeutic agent is or includes a cancer drug. In some embodiments, the second therapeutic agent is or includes a chemotherapeutic agent. In some embodiments, the second therapeutic agent is or includes a hormone therapy agent. In some embodiments, the second therapeutic agent is or includes a kinase inhibitor. In some embodiments, the second therapeutic agent is or includes a checkpoint inhibitor (e.g., an antibody against PD-1, PD-L1, CTLA-4, etc.). In some embodiments, provided agents can be administered at a lower unit dose and / or total dose compared to when used alone. In some embodiments, the second agent can be administered at a lower unit dose and / or total dose compared to when used alone. In some embodiments, one or more side effects associated with administration of the provided agent and / or second therapy or therapeutic agent are reduced. In some embodiments, the combination therapy provides improved results, e.g., compared to when each agent is utilized individually. In some embodiments, the combination therapy achieves one or more better results, e.g., compared to when each agent is utilized individually.

[0036] Further description of certain embodiments of the provided technology is provided below. [Brief explanation of the drawings]

[0037] [Figure 1-1] The provided technology can selectively inhibit beta-catenin-driven gene transcription in beta-catenin-expressing cells. Stapled peptides inhibited endogenous gene expression in wild-type HAP1 isogenic cells but not in CTNNB1 knockout (KO) cells. (A): Beta-catenin levels. CHIR:CHIR99021, which can activate the beta-catenin pathway and increase AXIN2 and SP5 expression. (B): SP5 expression (24 hours). (C): AXIN2 expression (24 hours). From left to right, the groups are DMSO ("0" and "0"), peptide A (1 and 5 μM), I-66 (1 and 5 μM), and I-470 (1 and 5 μM). Expression was assessed after 24 hours of treatment. [Figure 1-2] Same as above.

[0038] [Figure 2] The provided technique is capable of reducing nuclear beta-catenin levels. Results for total beta-catenin in the nuclear fraction (24 hours) are shown as an example.

[0039] [Figure 3-1] The provided techniques can inhibit cell proliferation, modulate transcription, and / or induce cell cycle arrest. (A): The provided techniques can reduce cell proliferation. (B) and (C): The provided techniques can modulate gene expression. (B): AXIN 24 hours. (C): CXCL12 24 hours. (D): The provided techniques can induce cell cycle arrest. From left to right: Peptide A (1, 5, and 10 μM), I-66 (1, 5, and 10 μM), I-470 (1, 5, and 10 μM), and DMSO. [Figure 3-2] Same as above.

[0040] [Figure 4] The provided technology can provide robust dose-dependent antitumor effects in vivo. Both dose levels evaluated provided robust reductions in tumor size, with higher dose levels providing greater reductions. COLO320DM cells (colon cancer, mutations: APC, TP53) were utilized for the data presented. The top row is for vehicle treatment, the middle row is for I-66, 30 mg / kg, Q4D, and the bottom row is for I-66, 75 mg / kg, Q4D.

[0041] [Figure 5-1] The provided technology can provide sustained tumor exposure, a favorable pharmacokinetic profile, and broad tissue distribution. (A): Sustained COLO320DM xenograft tumor exposure after a single IP injection of 50 mg / kg I-66 is shown as an example. The dotted line indicates the in vitro growth IC50 (0.7 μM). (B): Mouse plasma pharmacokinetics. Data shown are the plasma concentration (ng / mL) of I-66 over time as an example. (C): Tissue distribution observed for I-66 in one evaluation. Mouse single IP dose, 50 mg / kg. For each sample, the left column is the 24-hour data, and the right column is the 96-hour data. [Figure 5-2] Same as above.

[0042] [Figure 6] 1H NMR (DMSO-d6, 373K) of a preparation of I-66 prepared as described in Example 9.

[0043] [Figure 7] Integration of peaks in the H NMR spectrum (DMSO-d, 373 K) of a preparation of I-66 prepared as described in Example 9. It will be understood by those skilled in the art that the integration may be further adjusted and / or optimized.

[0044] [Figure 8] The provided technology can provide robust anti-tumor efficacy in vivo in multiple tumor models. (A): Selected data from a PDX colon cancer model. (B): Selected data from a PDX CRC model. DETAILED DESCRIPTION OF THE INVENTION

[0045] Detailed Description of Certain Embodiments definition As used herein, the following definitions shall apply unless otherwise specified: For the purposes of this disclosure, chemical elements are defined as defined in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Further, the general principles of organic chemistry are identified in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5 th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001.

[0046] Administration: As used herein, the term "administration" typically refers to the administration of a composition to a subject or system. Those of skill in the art will recognize various routes that can be utilized for administration to a subject, e.g., a human, in appropriate circumstances. For example, in some embodiments, administration can be intraocular, oral, parenteral, topical, etc. In certain embodiments, administration can be bronchial (e.g., by intrabronchial instillation), buccal, cutaneous (e.g., can be or include one or more of topical application to the dermis, intradermal, transdermal, transdermal, etc.), enteral, intraarterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, intraspecific organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), intravaginal, intravitreal, etc. In some embodiments, administration may involve intermittent dosing (e.g., multiple doses separated in time) and / or periodic dosing (e.g., individual doses separated by a common period of time). In some embodiments, administration may involve continuous dosing (e.g., perfusion) over at least a selected period of time.

[0047] Affinity: As known in the art, "affinity" is a measure of the tightness of binding of a particular ligand (e.g., a drug) to its partner (e.g., beta-catenin or a portion thereof). Affinity can be measured in different ways. In some embodiments, affinity is measured by a quantitative assay. In some such embodiments, the concentration of the binding partner can be fixed in excess of the concentration of the ligand to mimic physiological conditions. Alternatively or additionally, in some embodiments, the concentration of the binding partner and / or the concentration of the ligand can be varied. In some such embodiments, affinity can be compared to a reference under equivalent conditions (e.g., concentrations).

[0048] Agent: In general, the term "agent," as used herein, may be used to refer to any chemical class of compound or entity, including, for example, polypeptides, nucleic acids, saccharides, lipids, small molecules, metals, or combinations or complexes thereof. In appropriate circumstances, as will be clear to one of skill in the art from the context, the term may be used to refer to an entity that is or includes a cell or organism, or a fraction, extract, or component thereof. Alternatively, or in addition, as will be clear from the context, the term may be used to refer to a natural product found in nature and / or obtained from nature. In some cases, again, as will be clear from the context, the term may be used to refer to one or more entities that are man-made in that they have been designed, engineered, and / or created through the action of the human hand, and / or are not found in nature. In some embodiments, agents may be used in isolated or pure form; in some embodiments, agents may be used in crude form. In some embodiments, potential agents may be provided as collections or libraries that, for example, can be screened to identify or characterize active agents therein. In some cases, the term "drug" can refer to a compound or entity that is or comprises a polymer; in some cases, the term can refer to a compound or entity that comprises one or more polymer moieties. In some embodiments, the term "drug" can refer to a compound or entity that is not a polymer and / or that is substantially free of any polymer and / or one or more specific polymer moieties. In some embodiments, the term can refer to a compound or entity that lacks or is substantially free of any polymer moieties. In some embodiments, the drug is a compound. In some embodiments, the drug is a stapled peptide.

[0049] Aliphatic: As used herein, "aliphatic" refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring (but is not aromatic) that is fully saturated or contains one or more units of unsaturation, or a combination thereof. In some embodiments, an aliphatic group contains 1-50 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-20 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-10 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-9 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-8 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-7 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms. In still other embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms, and in yet other embodiments, aliphatic groups contain 1, 2, 3, or 4 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0050] Alkenyl: As used herein, the term "alkenyl" refers to an aliphatic group, as defined herein, having one or more double bonds.

[0051] Alkyl: As used herein, the term "alkyl" is given its ordinary meaning in the art and can include saturated aliphatic groups, including straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and cycloalkyl-substituted alkyl groups. In some embodiments, an alkyl has 1 to 100 carbon atoms. In certain embodiments, a straight-chain or branched-chain alkyl has about 1 to 20 carbon atoms in its backbone (e.g., C1 to C6 for straight chain). 20 , and for branched chains, C2 to C 20 ), or about 1-10 carbon atoms. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure, be it monocyclic, bicyclic or polycyclic, or about 5, 6 or 7 carbons in the ring structure. In some embodiments, alkyl groups can be lower alkyl groups, which lower alkyl groups have from 1-4 carbon atoms (e.g., C1-C4 for a straight chain lower alkyl).

[0052] Amino Acid: As used herein in its broadest sense, refers to any compound and / or substance that can be incorporated into a polypeptide chain, for example, by the formation of one or more peptide bonds. In some embodiments, an amino acid comprises an amino group and a carboxylic acid group. In some embodiments, an amino acid comprises an NH(R a1 )-L a1 -C(R a2 )(R a3 )-L a2In some embodiments, an amino acid has the general structure NH(R')-C(R')-COOH, where each R' is independently as described in this disclosure. In some embodiments, an amino acid has the general structure HN-C(R')-COOH, where R' is as described in this disclosure. In some embodiments, an amino acid has the general structure HN-C(H)(R')-COOH, where R' is as described in this disclosure. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid. A "standard amino acid" refers to any of the 20 standard L-amino acids commonly found in naturally occurring peptides. A "non-standard amino acid" refers to any amino acid other than the standard amino acids, whether prepared synthetically or obtained from a natural source. In some embodiments, amino acids, including the carboxy- and / or amino-terminal amino acids of a polypeptide, may contain structural modifications compared to the general structure described above. For example, in some embodiments, amino acids may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of an amino group, a carboxylic acid group, one or more protons, one or more hydrogens, and / or a hydroxyl group) compared to the general structure. In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide containing a modified amino acid compared to a polypeptide containing an otherwise identical, unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide containing a modified amino acid compared to a polypeptide containing an otherwise identical, unmodified amino acid.As will be clear from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments, the term may be used to refer to an amino acid residue of a polypeptide.

[0053] Analog: As used herein, the term "analog" refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an "analog" exhibits significant structural similarity to the reference substance, e.g., shares a core or consensus structure, but differs in certain distinct respects. In some embodiments, an analog is a substance that can be produced from a reference substance, e.g., by chemically manipulating the reference substance. In some embodiments, an analog is a substance that can be produced by performing a synthetic process that is substantially similar to (e.g., shares multiple steps with) one that produces the reference substance. In some embodiments, an analog is produced or can be produced by performing a synthetic process that is different from that used to produce the reference substance.

[0054] Animal: As used herein, refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans of either sex and at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, and / or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and / or a clone.

[0055] Approximately: As used herein, the term "approximately" or "about," when applied to one or more values ​​of interest, refers to a value similar to a stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less percent of a stated reference value in either direction (greater than or less than the reference value), unless otherwise specified or clear from the context (except when such number exceeds 100% of the possible values).

[0056] Aryl: The term "aryl," used alone or as part of a larger moiety, such as in "aralkyl," "aralkoxy," "aryloxyalkyl," etc., refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, where at least one ring in the system is aromatic. In some embodiments, an aryl group is a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 14 ring members, where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. In some embodiments, an aryl group is a biaryl group. The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments of the present disclosure, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracyl, etc., which may bear one or more substituents. In some embodiments, groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, are also included within the scope of the term "aryl" as used herein, where the radical or point of attachment is on the aryl ring.

[0057] Associated with: As used herein, two events or entities are "associated" with each other when the existence, level and / or form of one correlates with the existence, level and / or form of the other.For example, a particular entity (e.g., nucleic acid (e.g., genomic DNA, transcript, mRNA, etc.), polypeptide, gene signature, metabolite, microorganism, etc.) is considered to be associated with a particular disease, disorder, or condition when its existence, level and / or form correlates with the incidence and / or susceptibility of the disease, disorder, or condition (e.g., across a relevant population).

[0058] Binding: As used herein, the term "binding" will be understood to typically refer to a non-covalent association between agents (two or more). In many embodiments herein, binding is addressed with respect to a particular agent and beta-catenin. Those skilled in the art will understand that such binding can be assessed in any of a variety of contexts. In some embodiments, binding is assessed with respect to beta-catenin. In some embodiments, binding is assessed with respect to one or more amino acid residues of beta-catenin. In some embodiments, binding is assessed with respect to one or more amino acid residues that correspond to amino acid residues of beta-catenin (e.g., are similarly positioned in three-dimensional space and / or have certain similar properties and / or functions).

[0059] Binding site: The term "binding site," as used herein, refers to a region of a target polypeptide that is formed in three-dimensional space and that includes one or more or all of the interacting residues of the target polypeptide. In some embodiments, a "binding site" may refer to one or more amino acid residues that include or are one or more or all of the interacting amino acid residues of the target polypeptide. As will be understood by those skilled in the art, binding sites may include residues that are adjacent to each other in the linear chain and / or residues that are distal to each other in the linear chain but approach each other in three-dimensional space when the target polypeptide is folded. Binding sites may include amino acid residues and / or saccharide residues.

[0060] Carrier: As used herein, refers to a diluent, adjuvant, excipient, or vehicle with which a composition is administered. In some exemplary embodiments, a carrier may comprise a sterile liquid, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. In some embodiments, a carrier is or comprises one or more solid components.

[0061] Equivalent: As used herein, the term "equivalent" refers to two or more agents, entities, circumstances, sets of conditions, etc. that are not necessarily identical to each other, but are sufficiently similar that a person skilled in the art would understand that a comparison between them is possible and that conclusions can therefore be reasonably drawn based on the observed differences or similarities. In some embodiments, equivalent sets of conditions, circumstances, individuals, or populations are characterized by multiple substantially identical traits and one or a few varying traits. The degree of identity required for two or more such agents, entities, circumstances, sets of conditions, etc. to be considered equivalent in any given situation will be understood by one skilled in the art depending on the situation. For example, a person skilled in the art will understand that sets of circumstances, individuals, or populations are equivalent to each other if they are characterized by a sufficient number and type of substantially identical traits to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicate variations in those varying traits.

[0062] Composition: Those skilled in the art will understand that the term "composition" can be used to refer to a separate physical entity that includes one or more specified components. Generally, unless otherwise specified, a composition can be in any form, e.g., gas, gel, liquid, solid, etc.

[0063] Alicyclic: As used herein, the term "alicyclic" refers to a saturated or partially unsaturated aliphatic monocyclic, bicyclic, or polycyclic ring system, e.g., having 3 to 30 members, where the aliphatic ring system is optionally substituted. Alicyclic groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, cycloalkyl has 3 to 6 carbons. The term "alicyclic" can also include an aliphatic ring fused to one or more aromatic or non-aromatic rings, such as decahedronaphthyl or tetrahydronaphthyl, where the radical or point of attachment is on the aliphatic ring. In some embodiments, a carbocyclic group is bicyclic. In some embodiments, a carbocyclic group is tricyclic. In some embodiments, a carbocyclic group is polycyclic. In some embodiments, an "alicyclic" (or "carbocyclic" or "cycloalkyl") is a monocyclic C-C 10 or C3-C6 hydrocarbons, or C4-C6 hydrocarbons that are fully saturated or contain one or more unsaturated units but are not aromatic 10 Or C8~C 10 Bicyclic hydrocarbons or C9-C that are fully saturated or contain one or more unsaturated units but are not aromatic 16 Refers to tricyclic hydrocarbons.

[0064] Derivative: As used herein, the term "derivative" refers to a structural analog of a reference substance. That is, a "derivative" is a substance that exhibits significant structural similarity to the reference substance, e.g., shares a core or consensus structure, but differs in certain distinct respects. In some embodiments, a derivative is a substance that can be produced from the reference substance by chemical manipulation. In some embodiments, a derivative is a substance that can be produced by performing a synthetic process substantially similar to (e.g., sharing multiple steps with) that produces the reference substance.

[0065] Dosage form or unit dosage form: Those of skill in the art will understand that the term "dosage form" can be used to refer to a physically discrete unit of active agent (e.g., a therapeutic or diagnostic agent) for administration to a subject. Typically, each such unit contains a predetermined quantity of active agent. In some embodiments, such quantity is a unit dosage (or an entire fraction thereof) suitable for administration according to a dosing regimen determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., using a therapeutic dosing regimen). Those of skill in the art will understand that the total amount of a therapeutic composition or agent to be administered to a particular subject is determined by one or more attending physicians and may include administration of multiple dosage forms.

[0066] Dosing regimen: Those skilled in the art will understand that the term "dosing regimen" can be used to refer to a set of unit doses (typically more than one) that are administered separately, typically spaced apart. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which can include one or more dosings. In some embodiments, a dosing regimen includes multiple dosings, each separated in time from the other dosings. In some embodiments, the individual dosings are separated from each other by periods of equal length; in some embodiments, a dosing regimen includes multiple dosings, each separated by at least two different periods. In some embodiments, all dosings within a dosing regimen are administered at the same unit dose. In some embodiments, different dosings within a dosing regimen are administered in different amounts. In some embodiments, a dosing regimen includes a first dosing at a first dosage, followed by one or more additional dosings at a second dosage that is different from the first dosage. In some embodiments, the dosing regimen includes a first dosing at a first dosage amount, followed by one or more additional dosings at a second dosage amount that is the same as the first dosage amount. In some embodiments, the dosing regimen is one that correlates with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).

[0067] Engineered: In general, the term "engineered" refers to aspects that have been manipulated by the hand of man. For example, in some embodiments, a peptide can be considered engineered if its amino acid sequence was selected by a human. For example, an engineered agent has an amino acid sequence selected based on preferences for corresponding amino acids at specific sites in protein-protein interactions. In some embodiments, the engineered sequence has an amino acid sequence that differs from the amino acid sequence of a polypeptide contained in the NCBI database that binds to the TCF site of beta-catenin. In many embodiments, the agent provided is an engineered agent. In some embodiments, the engineered agent is a peptide agent comprising non-natural amino acid residues, non-natural amino acid sequences, and / or peptide staples. In some embodiments, the agent provided comprises or is an engineered peptide agent comprising an engineered sequence.

[0068] Halogen: The term "halogen" means F, Cl, Br, or I.

[0069] Heteroaliphatic: The term "heteroaliphatic" is given its ordinary meaning in the art and refers to an aliphatic group, as described herein, in which one or more carbon atoms are replaced by one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.).

[0070] Heteroalkyl: The term "heteroalkyl" is given its ordinary meaning in the art and refers to an alkyl group, as described herein, in which one or more carbon atoms have been replaced with a heteroatom (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). Examples of heteroalkyl groups include, but are not limited to, alkoxy, poly(ethylene glycol)-, alkyl-substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, and the like.

[0071] Heteroaryl: The terms "heteroaryl" and "heteroar-," used alone or as part of a larger moiety, e.g., "heteroaralkyl" or "heteroaralkoxy," refer to a monocyclic, bicyclic, or polycyclic ring system having, for example, a total of 5 to 30 ring members, e.g., 5, 6, 9, 10, 14, etc., wherein at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, heteroaryl groups are groups having 5 to 10 ring atoms (i.e., monocyclic, bicyclic, or polycyclic), and in some embodiments, groups having 5, 6, 9, or 10 ring atoms. In some embodiments, heteroaryl groups share 6, 10, or 14 pi electrons in a cyclic arrangement and have 1 to 5 heteroatoms in addition to the carbon atoms. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. In some embodiments, heteroaryl is a heterobiaryl group such as bipyridyl. The terms "heteroaryl" and "heteroar-," as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or polycyclic.The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," all of which terms encompass rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl group, where the alkyl and heteroaryl portions independently are optionally substituted.

[0072] Heteroatom: The term "heteroatom" means an atom that is neither carbon nor hydrogen. In some embodiments, a heteroatom is oxygen, sulfur, nitrogen, phosphorus, boron, or silicon (any oxidized form of nitrogen, sulfur, phosphorus, or silicon; any basic or substitutable nitrogen of a heterocycle (e.g., N, such as in 3,4-dihydro-2H-pyrrolyl), NH (such as in pyrrolidinyl), or NR + (including quaternized forms of N-substituted pyrrolidinyl); etc.). In some embodiments, the heteroatom is boron, nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur.

[0073] Heterocyclyl: As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocycle" are used interchangeably and refer to a monocyclic, bicyclic, or polycyclic ring moiety (e.g., 3-30 members) that is saturated or partially unsaturated and has one or more heteroatom ring atoms. In some embodiments, the heteroatom is boron, nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, silicon, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, sulfur, or phosphorus. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, the heterocyclyl group is a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety that is saturated or partially unsaturated and has, in addition to carbon atoms, one or more, preferably 1-4, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen may be N (such as in 3,4-dihydro-2H-pyrrolyl), NH (such as in pyrrolidinyl), or +It can be NR (such as in N-substituted pyrrolidinyl). The heterocycle can be attached to the pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heteroaliphatic ring. Heterocyclyl groups can be monocyclic, bicyclic, or polycyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl portions independently are optionally substituted.

[0074] Homology: As used herein, the term "homology" refers to the overall relatedness between polymer molecules, e.g., between nucleic acid molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polymer molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, polymer molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., contain residues with related chemical properties at corresponding positions). For example, as is well known to those skilled in the art, certain amino acids are typically classified as "hydrophobic" or "hydrophilic" amino acids, and / or as having "polar" or "non-polar" side chains, similar to one another. Substitution of one amino acid for another of the same type can often be considered a "homologous" substitution. Typical amino acid categorizations are summarized below (hydrophobicity scale of Kyte and Doolittle, 1982: A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 157: 105-132): [Table 1]

[0075] As those skilled in the art will understand, various algorithms are available that allow sequences to be compared to determine the degree of sequence homology, including allowing a gap of a specified length in one sequence compared to another sequence when considering which residues in different sequences "correspond" to each other.The percent homology between two nucleic acid sequences can be calculated, for example, by aligning the two sequences for optimal comparison (e.g., for comparison, gaps can be introduced into one or both of the first nucleic acid sequence and the second nucleic acid sequence for optimal alignment, and non-corresponding sequences can be ignored).In certain embodiments, the length of the sequence aligned for comparison is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of the reference sequence.Then, the nucleotides at corresponding nucleotide positions are compared. If a position in a first sequence is occupied by the same nucleotide as the corresponding position in a second sequence, the molecules are identical at that position; if a position in a first sequence is occupied by a nucleotide similar to the corresponding position in a second sequence, the molecules are similar at that position. The percent homology between two sequences is a function of the number of identical and similar positions shared by the sequences, taking into account the number of gaps and the length of each gap that need to be introduced to optimally align the two sequences. Representative algorithms and computer programs useful in determining the percent homology between two nucleotide sequences include, for example, the Meyers and Miller algorithm (CABIOS, 1989, 4: 11-17), which is incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.Alternatively, the percent homology between two nucleotide sequences can be determined using, for example, the GAP program from the GCG software package, using the NWSgapdna.CMP matrix.

[0076] Interacting residues: The terms "interacting residues" and "interacting motifs," as used herein, refer to residues or motifs of a drug that are designed to interact with a specific target residue in a target polypeptide, or to residues of a target polypeptide that interact with a specific motif (e.g., aromatic group, amino acid residue, etc.) of a drug. Specifically, various interacting residues and motifs of a drug are selected and positioned within the drug (e.g., during binding, docking, or other interaction assays) so as to be displayed in three-dimensional space within a predetermined distance (or volume) relative to the identified target residue. In many embodiments, the interacting residues are direct-binding residues.

[0077] "Improved," "increased," or "reduced": As used herein, these terms, or grammatically equivalent comparative terms, refer to a value compared to a comparable reference measurement. For example, in some embodiments, an assessment achieved with an agent of interest may be "improved" compared to an assessment obtained with an equivalent reference agent. Alternatively, or in addition, in some embodiments, an assessment achieved in a subject or system of interest may be "improved" compared to an assessment obtained in the same subject or system under different conditions (e.g., before or after an event such as administration of the agent of interest) or in a different comparable subject (e.g., in a comparable subject or system different from the subject or system of interest in which one or more indicators of a particular disease, disorder, or condition of interest are present or have previously been exposed to a condition, agent, etc.). In some embodiments, comparative terms refer to a statistically relevant difference (e.g., one of sufficient prevalence and / or magnitude to achieve statistical relevance). Those skilled in the art will recognize or can readily determine the degree and / or prevalence of difference necessary or sufficient to achieve such statistical significance in a given situation.

[0078] Partially unsaturated: As used herein, the term "partially unsaturated" refers to a moiety that contains at least one double or triple bond. The term "partially unsaturated" is intended to encompass groups with multiple sites of unsaturation, but is not intended to encompass aryl or heteroaryl moieties.

[0079] Peptide: The term "peptide," as used herein, refers to a polypeptide. In some embodiments, a peptide is a polypeptide having a length of, for example, less than about 100 amino acids, less than about 50 amino acids, less than about 40 amino acids, less than about 30 amino acids, less than about 25 amino acids, less than about 20 amino acids, less than about 15 amino acids, or less than 10 amino acids. In some embodiments, the length is about 5-20, 5-19, 5-18, 5-17, 5-16, 5-15, 10-20, 10-19, 10-18, 10-17, 10-16, 10-15, 11-20, 11-19, 11-18, 11-17, 11-16, 11-15, 12-20, 12-19, 12-18, 12-17, 12-16, 12 up to 15, 13-20, 13-19, 13-18, 13-17, 13-16, 13-15, 14-20, 14-19, 14-18, 14-17, 14-16, 14-15, or about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids.

[0080] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose suitable for administration in a therapeutic regimen that, when administered to a relevant population, exhibits a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, the pharmaceutical compositions can be specially formulated for administration in solid or liquid form, including those adapted for: oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as a sterile solution or suspension, or sustained-release formulation; topical application, e.g., as a cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; vaginal or rectal, e.g., as a suppository, cream, or foam; sublingual; intraocular; transdermal; or intranasal, pulmonary, and other mucosal surfaces.

[0081] Pharmaceutically acceptable: As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms which, within the scope of good medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0082] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that carries or transports a compound of interest from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository wax; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and dairy. glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic, compatible substances used in pharmaceutical formulations.

[0083] Pharmaceutically acceptable salts: The term "pharmaceutically acceptable salts," as used herein, refers to salts of such compounds that are suitable for use in a pharmaceutical context, i.e., salts that are within the scope of good medical judgment, suitable for use in contact with the tissues of humans and lower animals, without undue toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known. For example, SM Berge, et al., J. Pharmaceutical Sciences, 66: 1-19 (1977), describes pharmaceutically acceptable salts in detail. In some embodiments, pharmaceutically acceptable salts include non-toxic acid addition salts, which are salts of amino groups formed with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other known methods, such as ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxybenzoate, benzoic acid, benzoyl benzoate ... Examples of the salts include dimethylsulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate.In some embodiments, pharmaceutically acceptable salts include non-toxic base addition salts, such as, but not limited to, those formed with bases by acidic groups of the provided compounds. Representative alkali metal or alkaline earth metal salts include sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, and the like. In some embodiments, pharmaceutically acceptable salts include ammonium salts (e.g., -N(R)3). + ) In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed with counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and arylsulfonates.

[0084] Polypeptide: As used herein, refers to any polymeric chain of amino acids. In some embodiments, a polypeptide has an amino acid sequence that occurs in nature. In some embodiments, a polypeptide has an amino acid sequence that does not occur in nature. In some embodiments, a polypeptide has an amino acid sequence that is engineered, in that it has been designed and / or created through the action of the hand of man. In some embodiments, a polypeptide may comprise or consist of natural amino acids, unnatural amino acids, or both. In some embodiments, a polypeptide may comprise or consist of only natural amino acids or only unnatural amino acids. In some embodiments, a polypeptide may comprise D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino acids. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may comprise one or more pendant groups or other modifications modifying or attached to one or more amino acid side chains, for example, at the N-terminus of the polypeptide, at the C-terminus of the polypeptide, or any combination thereof. In some embodiments, such pendant groups or modifications may be selected from the group consisting of acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations. In some embodiments, a polypeptide may be cyclic and / or may include a cyclic portion. In some embodiments, a polypeptide is not cyclic and / or does not include a cyclic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or include a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to the name of a reference polypeptide, activity, or structure; in such cases, the term is used herein to refer to polypeptides that share a related activity or structure and therefore can be considered members of the same class or family of polypeptides.For each such class, exemplary polypeptides falling within the class for which amino acid sequence and / or function are known are presented herein and / or will be recognized by those of skill in the art; in some embodiments, such exemplary polypeptides are the reference polypeptides of the class or family of polypeptides. In some embodiments, members of a class or family of polypeptides exhibit significant sequence homology or identity with the reference polypeptides of that class, and in some embodiments, with all polypeptides within that class, share common sequence motifs (e.g., characteristic sequence elements), and / or share a common activity (in some embodiments, at comparable levels or within a specified range)). For example, in some embodiments, member polypeptides exhibit a degree of overall sequence homology or identity with a reference polypeptide of at least about 30-40%, often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater, and / or contain at least one region (e.g., a conserved region which, in some embodiments, may be or may contain a distinctive sequence element) that exhibits very high sequence identity, often greater than 90%, or even 95%, 96%, 97%, 98%, or 99%. Such conserved regions typically encompass at least 3-4 amino acids, and often up to 20 or more amino acids; in some embodiments, the conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more contiguous amino acids. In some embodiments, the related polypeptide may comprise or consist of a fragment of a parent polypeptide.In some embodiments, a useful polypeptide may comprise or consist of multiple fragments, each of which is found in the same parent polypeptide in a different spatial arrangement relative to one another than is found in the polypeptide of interest (e.g., fragments that are directly linked in the parent may be spatially separated in the polypeptide of interest, or vice versa, and / or fragments may be present in a different order in the polypeptide of interest than in the parent), and thus the polypeptide of interest is a derivative of its parent polypeptide.

[0085] Prevent or prophylaxis: As used herein, when used in reference to the occurrence of a disease, disorder, and / or condition, refers to reducing the risk of the disease, disorder, and / or condition occurring and / or delaying the onset of one or more characteristics or symptoms of the disease, disorder, or condition. Prevention may be considered complete when the onset of the disease, disorder, or condition has been delayed for a predefined period of time.

[0086] Protecting Group: The term "protecting group" as used herein is a group that is well known in the art and is described in Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, 3 rdedition, John Wiley & Sons, 1999. Also included are protecting groups specifically adapted for nucleoside and nucleotide chemistry, as described in Current Protocols in Nucleic Acid Chemistry, edited by Serge L. Beaucage et al. 06 / 2012, the entirety of Chapter 2 of which is incorporated herein by reference. Suitable amino protecting groups include methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2- Phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-Dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p- Nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthio Phenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl Carbamates, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivatives, N'-p-toluenesulfonylaminocarbonyl derivatives, N'-phenylaminothiocarbonyl derivatives, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-Dimethoxycarbonylvinylcarbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isobornyl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclohexyl p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate acetamide, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinamide, N-acetylmethionine derivatives, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-Dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrroline (py (roolin)-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzyly N-phenyl-, Np-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-di Phenylborinic acid derivatives, N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidate, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-Dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridine sulfenamide (Npys), p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte) , 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0087] In some embodiments, suitable mono-protected amines include, but are not limited to, aralkylamines, carbamates, allylamines, amides, etc. Examples of suitable mono-protected amino moieties include t-butyloxycarbonylamino (-NHBOC), ethyloxycarbonylamino, methyloxycarbonylamino, trichloroethyloxycarbonylamino, allyloxycarbonylamino (-NHAlloc), benzyloxocarbonylamino (-NHCBZ), allylamino, benzylamino (-NHBn), fluorenylmethylcarbonyl (-NHFmoc), formamide, acetamide, chloroacetamide, dichloroacetamide, trichloroacetamide, phenylacetamide, trifluoroacetamide, benzamide, t-butyldiphenylsilyl, etc. In some embodiments, suitable di-protected amines include amines substituted with two substituents independently selected from those described above as mono-protected amines, and further include cyclic imides such as phthalimide, maleimide, succinimide, etc. In some embodiments, suitable di-protected amines include pyrroles, such as 2,2,5,5-tetramethyl-[1,2,5]azadisilolidine, and the like, and azides.

[0088] Suitable protected carboxylic acids further include, but are not limited to, silyl-protected carboxylic acids, alkyl-protected carboxylic acids, alkenyl-protected carboxylic acids, aryl-protected carboxylic acids, and arylalkyl-protected carboxylic acids. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, etc. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, and tetrahydropyran-2-yl. Examples of suitable alkenyl groups include allyl. Examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Examples of suitable arylalkyl groups include optionally substituted benzyl (e.g., p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl), and 2-picolyl and 4-picolyl. In some embodiments, suitable protected carboxylic acids include, but are not limited to, optionally substituted C 1~6 These include aliphatic esters, optionally substituted aryl esters, silyl esters, activated esters, amides, hydrazides, and the like. Examples of such ester groups include methyl esters, ethyl esters, propyl esters, isopropyl esters, butyl esters, isobutyl esters, benzyl esters, and phenyl esters, each of which is optionally substituted. Additional suitable protected carboxylic acids include oxazolines and orthoesters.

[0089] Suitable hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl ( MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]phenyl ]-4-Methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trimethyl- methylethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4''-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4''-tris(levulinoyloxyphenyl)methyl, 4,4',4''-tris(benzoyloxyphenyl)methyl methyl, 3-(imidazol-1-yl)bis(4',4''-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropyl Silyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxy Acetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyl dithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate Alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6 -dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). For the protection of 1,2-diols or 1,3-diols, the protecting groups include methylene acetal, ethylidene acetal, 1-t-butyl ethylidene ketal, 1-phenyl ethylidene ketal, (4-methoxyphenyl) ethylidene acetal, 2,2,2-Trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, 1-methoxyethylidene orthoester, 1-ethoxyethylidene orthoester, 1,2-dimethoxyethylidene Examples of suitable benzylidene orthoesters include ethylidene orthoesters, α-methoxybenzylidene orthoesters, 1-(N,N-dimethylamino)ethylidene derivatives, α-(N,N'-dimethylamino)benzylidene derivatives, 2-oxacyclopentylidene orthoesters, di-t-butylsilylene groups (DTBS), 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivatives (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivatives (TBDS), cyclic carbonates, cyclic boronates, ethyl boronate, and phenyl boronate.

[0090] In some embodiments, the hydroxyl protecting group is acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl(trityl), 4,4′-dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl. aryl, triphenylsilyl, triisopropylsilyl, benzoylformate, chloroacetyl, trichloroacetyl, trifluoroacetyl, pivaloyl, 9-fluorenylmethyl carbonate, mesylate, tosylate, triflate, trityl, monomethoxytrityl (MMTr), 4,4'-dimethoxytrityl (DMTr) and 4,4',4''-trimethoxytrityl (TMTr), 2-cyanoethyl (CE or Cne), 2-(trimethylsilyl)ethyl (TSE), 2-(2-nitrophenyl)ethyl, 2-(4-cyanophenyl)ethyl 2-(4-nitrophenyl)ethyl (NPE), 2-(4-nitrophenylsulfonyl)ethyl, 3,5-dichlorophenyl, 2,4-dimethylphenyl, 2-nitrophenyl, 4-nitrophenyl, 2,4,6-trimethylphenyl, 2-(2-nitrophenyl)ethyl, butylthiocarbonyl, 4,4',4"-tris(benzoyloxy)trityl, diphenylcarbamoyl, levulinyl, 2-(dibromomethyl)benzoyl (Dbmb), 2-(isopropylthiomethoxymethyl)benzoyl (Ptmt), 9-phenylxanthen-9-yl (pixyl), or 9-(p-methoxyphenyl)xanthine-9-yl (MOX). In some embodiments, each of the hydroxyl protecting groups is independently selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and 4,4'-dimethoxytrityl. In some embodiments, the hydroxyl protecting group is selected from the group consisting of trityl, monomethoxytrityl, and 4,4'-dimethoxytrityl groups.In some embodiments, the phosphite-linked protecting group is a group that is attached to the phosphite linkage (e.g., internucleotide linkage) throughout oligonucleotide synthesis. In some embodiments, the protecting group is attached to the sulfur atom of a phosphorothioate group. In some embodiments, the protecting group is attached to the oxygen atom of an internucleotide phosphorothioate linkage. In some embodiments, the protecting group is attached to the oxygen atom of an internucleotide phosphate linkage. In some embodiments, the protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2-sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2-(p-nitrophenyl)ethyl (NPE or Npe), 2-phenylethyl, 3-(N-tert-butylcarboxamido)-1-propyl, 4-oxopentyl, 4-methylthio-1-butyl, 2-cyano-1,1-dimethylethyl, 4-N-methylaminobutyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, 2-(N-formyl,N-methyl)aminoethyl, or 4-[N-methyl-N-(2,2,2-trifluoroacetyl)amino]butyl.

[0091] Protected thiols are well known in the art and include those described in detail in Greene (1999). Suitable protected thiols further include, but are not limited to, disulfides, thioethers, silyl thioethers, thioesters, thiocarbonates, and thiocarbamates. Examples of such groups include, but are not limited to, alkyl thioethers, benzyl and substituted benzyl thioethers, triphenylmethyl thioethers, and trichloroethoxycarbonyl thioesters, to name a few.

[0092] Reference: As used herein, describes a standard or control against which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially simultaneously with the test or determination of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as will be understood by those skilled in the art, a reference or control is determined or characterized under conditions or circumstances equivalent to those under evaluation. In each case, those skilled in the art will understand whether sufficient similarity exists to justify reliance on and / or comparison to a particular potential reference or control.

[0093] Specificity: As known in the art, "specificity" is a measure of the ability of a particular ligand (e.g., an agent) to distinguish its binding partner (e.g., beta-catenin) from other potential binding partners (e.g., another protein, another portion (e.g., domain) of beta-catenin).

[0094] Substituted: As described herein, compounds of the present disclosure may be optionally substituted and / or contain substituted moieties. In general, the term "substituted," whether preceded by the term "optionally" or not, means that one or more hydrogens of the specified moiety have been replaced with a suitable substituent. Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and more than one position in any given structure may be substituted with more than one substituent selected from the specified group, and the substituents may be the same or different at all positions. Combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that remains substantially unchanged when subjected to conditions that permit its preparation, detection, and, in certain embodiments, recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, exemplary substituents are as follows:

[0095] Preferred monovalent substituents are halogen; -(CH2) 0~4 R°;-(CH2) 0~4 OR°;-O(CH2) 0~4 R o , -O-(CH2) 0~4 C(O)OR°;-(CH2) 0~4 CH(OR°)2; optionally substituted with R° -(CH2) 0~4 Ph; optionally substituted with R° -(CH2) 0~4 O(CH2) 0~1 Ph; optionally substituted with R° -CH=CHPh; optionally substituted with R° -(CH2) 0~4 O(CH2) 0~1 -pyridyl; -NO2; -CN; -N3; ​​-(CH2) 0~4 N(R°)2;-(CH2) 0~4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0~4 N(R°)C(O)N(R°)2;-N(R°)C(S)N(R°)2;-(CH2) 0~4N(R°)C(O)OR°;-N(R°)N(R°)C(O)R°;-N(R°)N(R°)C(O)N(R°)2;-N(R°)N(R°)C(O)OR°;-(CH2) 0~4 C(O)R°;-C(S)R°;-(CH2) 0~4 C(O)OR°;-(CH2) 0~4 C(O)SR°;-(CH2) 0~4 C(O)OSi(R°)3;-(CH2) 0~4 OC(O)R°;-OC(O)(CH2) 0~4 SR°, -SC(S)SR°;-(CH2) 0~4 SC(O)R°;-(CH2) 0~4 C(O)N(R°)2;-C(S)N(R°)2;-C(S)SR°;-SC(S)SR°, -(CH2) 0~4 OC(O)N(R°)2;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH2C(O)R°;-C(NOR°)R°;-(CH2) 0~4 SSR°;-(CH2) 0~4 S(O)2R°;-(CH2) 0~4 S(O)2OR°;-(CH2) 0~4 OS(O)2R°;-S(O)2N(R°)2;-(CH2) 0~4 S(O)R°;-N(R°)S(O)2N(R°)2;-N(R°)S(O)2R°;-N(OR°)R°;-C(NH)N(R°)2;-Si(R°)3;-OSi(R°)3;-P(R°)2;-P(OR °)2;-OP(R°)2;-OP(OR°)2;-N(R°)P(R°)2;-B(R°)2;-OB(R°)2;-P(O)(R°)2;-OP(O)(R°)2;-N(R°)P(O)(R°)2;-(C 1~4 Linear or branched alkylene)ON(R°)2; or -(C 1~4 straight or branched chain alkylene)C(O)ON(R°); where each R° is optionally substituted as defined below and independently represents hydrogen, C 1~20 Aliphatic, C having 1 to 5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus 1~20 Heteroaliphatic, -CH2-(C 6~14 aryl), -O(CH2)0~1 (C 6~14 aryl), -CH2- (5-14 membered heteroaryl ring), a 5-20 membered, mono-, bi-, or polycyclic, saturated, partially unsaturated, or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, or, notwithstanding the above definitions, two independently occurring R° together with their intervening atom(s) form a 5-20 membered, mono-, bi-, or polycyclic, saturated, partially unsaturated, or aryl ring having 0-5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, which may be substituted as defined below.

[0096] Suitable monovalent substituents on R° (or the ring formed by two independently occurring R° together with their intervening atoms) are independently halogen, —(CH) 0~2 R ● ,-(Halo R ● ), -(CH2) 0~2 OH, -(CH2) 0~2 OR ● , -(CH2) 0~2 CH(OR ● )2;-O(HaloR ● ), -CN, -N3, -(CH2) 0~2 C(O)R ● , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR ● , -(CH2) 0~2 SR ● , -(CH2) 0~2 SH, -(CH2) 0~2 NH2, -(CH2) 0~2 NHR ● , -(CH2) 0~2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● -(C 1~4 Straight or branched chain alkylene)C(O)OR ● , or -SSR ● where each R● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents on a saturated carbon atom of R° include =0 and =S.

[0097] Preferred divalent substituents are: =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2~3 O-, or -S(C(R * 2)) 2~3 S-, where each independently occurring R * each of which is hydrogen, optionally substituted as defined below, C 1~6 aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. Suitable divalent substituents attached to adjacent substitutable carbons of an "optionally substituted" group include -O(CR * 2) 2~3 O—, wherein each independently occurring R * each of which is hydrogen, optionally substituted as defined below, C 1~6 aliphatic, or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0098] R * Suitable substituents on the aliphatic groups are halogen, -R ● ,-(Halo R ● ), -OH, -OR● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0099] In some embodiments, a suitable substituent on a substitutable nitrogen is —R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † where each R † are independently hydrogen, optionally substituted C as defined below 1~6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or two independently occurring R † together with their intervening atom(s) form an unsubstituted 3-12 membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0100] R †Suitable substituents on the aliphatic group are independently halogen, —R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0101] Subject: As used herein, the term "subject" or "test subject" refers to any organism to which a provided compound or composition is administered in accordance with the present disclosure, e.g., for experimental, diagnostic, preventative, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and humans; insects; maggots; etc.) and plants. In some embodiments, the subject may be a subject suffering from and / or susceptible to a disease, disorder, and / or condition. In some embodiments, the subject is a human.

[0102] Susceptible: An individual who is "susceptible" to a disease, disorder, and / or condition is an individual who is at a higher risk than members of the general public of developing the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may be an individual who has not been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may be an individual who exhibits symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may be an individual who does not exhibit symptoms of the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may be an individual who develops the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition may be an individual who does not develop the disease, disorder, and / or condition.

[0103] Target polypeptide: A "target polypeptide," as that term is used herein, is a polypeptide with which an agent interacts. In some embodiments, the target polypeptide is a beta-catenin polypeptide. In some embodiments, the target polypeptide comprises, consists essentially of, or is a binding site for a beta-catenin polypeptide.

[0104] Target residue: A "target residue," as the term is used herein, is a residue in a target polypeptide with which an agent is designed to interact. For example, an agent may feature a particular interaction motif (e.g., an aromatic group as described herein) and / or residue (e.g., an amino acid residue containing an aromatic group as described herein), selected and positioned (by being displayed on a selected scaffold) to be within a certain predetermined distance (or volume) of the target residue. In some embodiments, the target residue is or includes an amino acid residue.

[0105] Therapeutic Agent: As used herein, the phrase "therapeutic agent" refers to an agent that, when administered to a subject, has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition.

[0106] Therapeutic regimen: "Therapeutic regimen," as that term is used herein, refers to a dosing regimen whose administration across a relevant population can be correlated with a desired or beneficial therapeutic outcome.

[0107] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response when administered as part of a treatment regimen. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, disorder, and / or condition when administered to a subject suffering from or susceptible to the disease, disorder, and / or condition. As will be understood by one of skill in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance being delivered, and the target cell or tissue. For example, an effective amount of a compound in a formulation to treat a disease, disorder, and / or condition is an amount that alleviates, ameliorate, relieves, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of, one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount.

[0108] Treat: As used herein, the terms "treat," "treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition. Treatment can be administered to subjects who do not show signs of the disease, disorder, and / or condition. In some embodiments, treatment can be administered to subjects who show only early signs of the disease, disorder, and / or condition, for example, to reduce the risk of developing pathologies associated with the disease, disorder, and / or condition.

[0109] Unit dose: As used herein, the phrase "unit dose" refers to an amount administered as a single dose and / or in a physically discrete unit of pharmaceutical composition. In many embodiments, a unit dose contains a predetermined quantity of an active agent. In some embodiments, a unit dose contains an entire single dose of an agent. In some embodiments, more than one unit dose is administered to achieve a total single dose. In some embodiments, administration of multiple unit doses is necessary or anticipated to be necessary to achieve the intended effect. A unit dose can be, for example, a volume of liquid (e.g., an acceptable carrier) containing a predetermined quantity of one or more therapeutic agents, a solid form of a predetermined amount of one or more therapeutic agents, a sustained-release formulation or drug delivery device containing a predetermined amount of one or more therapeutic agents, etc. It will be understood that a unit dose can exist as a formulation containing any of a variety of components in addition to the therapeutic agent(s). For example, acceptable carriers (e.g., pharmaceutically acceptable carriers), diluents, stabilizers, buffers, preservatives, etc., as described below, can be included. In many embodiments, the appropriate total daily dosage of a particular therapeutic agent may comprise a portion or a plurality of unit doses, and can be determined, for example, by an attending physician within the scope of good medical judgment, as will be understood by those skilled in the art. In some embodiments, the specific effective dose level for any particular subject or organism may depend on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific active compound used; the specific composition used; the age, weight, general health, sex, and diet of the subject; the administration time and excretion rate of the specific active compound used; the duration of treatment; drugs and / or additional therapies used in combination with or simultaneously with the specific compound(s) used, and similar factors well known in the medical field.

[0110] Unsaturated: The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation.

[0111] Unless otherwise specified, salts, such as pharmaceutically acceptable acid or base addition salts, stereoisomeric forms, and tautomeric forms of the compounds provided are included.

[0112] As used in this disclosure, unless otherwise clear from the context, (i) the term "a" or "an" may be understood to mean "at least one"; (ii) the term "or" may be understood to mean "and / or"; (iii) the terms "comprising," "comprise," "including" (whether or not used in conjunction with "limited to"), and "include" (whether or not used in conjunction with "limited to") may be understood to encompass the itemized components or steps, whether presented by themselves or with one or more additional components or steps; (iv) the term "another" may be understood to mean at least a certain additional / second one or more; (v) the terms "about" and "approximately" may be understood to allow for standard variations understood by those of ordinary skill in the art; and (vi) when ranges are presented, the endpoints are included. Staple Peptides

[0113] In some embodiments, the agent provided is or comprises a peptide. In some embodiments, the agent provided is a peptide. In some embodiments, the peptide is a stapled peptide. In some embodiments, the agent provided is a stapled peptide. In some embodiments, the peptide is a stitched peptide. In some embodiments, the agent provided is a stitched peptide. In some embodiments, the agent is a stitched peptide. In some embodiments, the stitched peptide comprises two or more staples, wherein the two staples are attached to the same peptide backbone atom. The stapled peptides described herein are typically peptides in which two or more amino acids in the peptide chain are linked by connection of two peptide backbone atoms of the amino acid residues, where the connection is not through the peptide backbone between the linked amino acid residues, as would be understood by one of skill in the art. In some embodiments, the staples described herein are linkers that link one amino acid residue to another amino acid residue, for example, by bonding to each of the peptide backbone atoms of the amino acid residues, where the connection by the staple is not through the peptide backbone between the linked amino acid residues, as would be understood by one of skill in the art. In some embodiments, the staples are attached to the peptide backbone by replacement of one or more hydrogens and / or substituents (e.g., side chains, O, S, etc.) on a peptide backbone atom (e.g., C, N, etc.). In some embodiments, the side chains form part of the staple. In some embodiments, the staples are attached to two carbon backbone atoms, e.g., two alpha carbon atoms. In some embodiments, the staples comprise C(R')2 or N(R'), either individually or as part of a larger moiety, where R' is R and can be R, with another group (e.g., R a3 ) together with their intervening atoms to form a ring as described herein (eg, when PyrS2 is stapled in various peptides).

[0114] In some embodiments, the staple peptide comprises one or more staples. In some embodiments, the staple peptide comprises two or more staples. In some embodiments, the staple peptide comprises three or more staples. In some embodiments, the staple peptide comprises four or more staples. In some embodiments, there are three staples in the staple peptide. In some embodiments, there are four staples in the staple peptide.

[0115] As will be appreciated by those skilled in the art, a variety of peptide stapling techniques, including both hydrocarbon- and non-hydrocarbon-stapling techniques, are available and can be utilized in accordance with the present disclosure. A variety of techniques for stapled and stitched peptides, including various staples and / or methods of making them, are available and can be utilized in accordance with the present disclosure, for example, those described in WO2019 / 051327 and WO2020 / 041270, the respective staples of which are incorporated herein by reference.

[0116] In some embodiments, the peptide, e.g., a stapled peptide, is or comprises a helical structure. In some embodiments, the peptide is a stapled peptide.

[0117] In some embodiments, the staple is a hydrocarbon staple. In some embodiments, the staple described herein is a non-hydrocarbon staple. In some embodiments, the non-hydrocarbon staple comprises one or more chain heteroatoms, wherein the staple chain is the shortest covalent connection within the staple from one end of the staple to the other end of the staple. In some embodiments, the non-hydrocarbon staple is or comprises at least one sulfur atom derived from an amino acid residue of a polypeptide. In some embodiments, the non-hydrocarbon staple comprises two sulfur atoms derived from two different amino acid residues of a polypeptide. In some embodiments, the non-hydrocarbon staple comprises two sulfur atoms derived from two different cysteine ​​residues of a polypeptide. In some embodiments, the staple is a cysteine ​​staple. In some embodiments, the staple is a non-cysteine ​​staple. In some embodiments, the non-hydrocarbon staple is a carbamate staple, comprising a carbamate moiety (e.g., -N(R')-C(O)-O-) within the chain. In some embodiments, the non-hydrocarbon staple is an amino staple and comprises an amino group (e.g., -N(R')-) in the chain. In some embodiments, the amino group of the amino staple, e.g., (-N(R')-), is not bonded to a carbon atom that further forms a double bond with a heteroatom (e.g., -C(=O), -C(=S), -C(=N-R'), etc.), and therefore is not part of another nitrogen-containing group such as an amide, carbamate, etc. In some embodiments, the non-hydrocarbon staple is an ester staple and comprises an ester moiety (-C(O)-O-) in the chain. In some embodiments, the non-hydrocarbon staple is an amide staple and comprises an amide moiety (-C(O)-N(R')-) in the chain. In some embodiments, the non-hydrocarbon staple is a sulfonamide staple and comprises a sulfonamide moiety (-S(O)-N(R')-) in the chain. In some embodiments, the non-hydrocarbon staple is an ether staple and comprises an ether moiety (-O-) in the chain.In some embodiments, R' of the carbamate moiety, amino group, amide moiety, sulfonamide moiety, or ether moiety is R, and R groups attached to the backbone (e.g., when R, R a3 ) and their intervening atoms together to form a ring as described herein. In some embodiments, R' of the carbamate moiety or amino group is R, and R groups attached to the backbone (e.g., when R, R a3 ) together with their intervening atoms to form a ring as described herein.

[0118] In some embodiments, the staple comprises one or more amino groups, e.g., —N(R′)—, where each R′ is independently as described herein. In some embodiments, —N(R′)— is bonded to two carbon atoms. In some embodiments, —N(R′)— is bonded to two carbon atoms, neither of which is bonded to any heteroatom by a double bond. In some embodiments, —N(R′)— is bonded to two sp3 carbon atoms. In some embodiments, the staple comprises one or more —C(O)—N(R′)— groups, where each R′ is independently as described herein. In some embodiments, the staple comprises one or more carbamate groups, e.g., one or more —(O)—C(O)—N(R′)—, where each R′ is independently as described herein. In some embodiments, R′ is —H. In some embodiments, R′ is an optionally substituted C 1~6 In some embodiments, R' is an optionally substituted C 1~6 In some embodiments, R' is C 1~6 In some embodiments, R' is C 1~6 In some embodiments, R' is methyl.

[0119] In some embodiments, the stapled peptide comprises one or more staples. In some embodiments, the stapled peptide comprises one and no more than one staple. In some embodiments, the stapled peptide comprises two and no more than two staples. In some embodiments, the two staples of the stapled peptide are attached to a common backbone atom. In some embodiments, the two staples of the stapled peptide are attached to a common backbone atom that is the alpha carbon atom of an amino acid residue. In some embodiments, the stapled peptide comprises three or more staples. In some embodiments, the stapled peptide comprises four or more staples. In some embodiments, the stapled peptide comprises three and no more than three staples. In some embodiments, the stapled peptide comprises four and no more than four staples. In some embodiments, each staple is independently selected from the group consisting of -L, -L- ... s1 -L s2 -L s3 In some embodiments, each staple is independently attached to two amino acid residues. In some embodiments, each staple is independently attached to two alpha carbon atoms.

[0120] In some embodiments, two, three, four, or all of the staples of the staple peptides are within a region having a length of several amino acid residues. In some embodiments, two staples are within such a region. In some embodiments, three staples are within such a region. In some embodiments, four staples are within such a region. In some embodiments, all staples are within such a region. In some embodiments, the region has a length of 5 to 20, 5 to 15, 5 to 14, 5 to 113, 5 to 12, 5 to 11, 5 to 10, 6 to 20, 6 to 15, 6 to 14, 6 to 113, 6 to 12, 6 to 11, 6 to 10, 7 to 20, 7 to 15, 7 to 14, 7 to 113, 7 to 12, 7 to 11, 7 to 10, 10 to 16, 10 to 15, 10 to 14, 11 to 16, 11 to 15, 11 to 14, 12 to 16, 12 to 15, 12 to 14, 13 to 15, or 13 to 14 amino acid residues. In some embodiments, the region has a length of 5 amino acid residues. In some embodiments, the region has a length of 6 amino acid residues. In some embodiments, the region is 7 amino acid residues in length. In some embodiments, the region is 8 amino acid residues in length. In some embodiments, the region is 9 amino acid residues in length. In some embodiments, the region is 10 amino acid residues in length. In some embodiments, the region is 11 amino acid residues in length. In some embodiments, the region is 12 amino acid residues in length. In some embodiments, the region is 13 amino acid residues in length. In some embodiments, the region is 14 amino acid residues in length. In some embodiments, the region is 15 amino acid residues in length. In some embodiments, the region is 16 amino acid residues in length. In some embodiments, the region is 17 amino acid residues in length. In some embodiments, the region is 18 amino acid residues in length. In some embodiments, the region is 19 amino acid residues in length. In some embodiments, the region is 20 amino acid residues in length.For example, in various embodiments, the staple peptide comprises three staples within a region of 14 amino acids (e.g., a staple attached to aa1 and aa4, a staple attached to aa4 and aa11, and a staple attached to aa10 and aa14).

[0121] In some embodiments, peptides, such as staple peptides of the present disclosure, are or include a helix structure. As will be appreciated by those skilled in the art, helices can have a variety of lengths. In some embodiments, the length of a helix ranges from 5 amino acid residues to 30 amino acid residues. In some embodiments, the length of a helix is ​​5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more amino acid residues. In some embodiments, the length of a helix is ​​6 amino acid residues. In some embodiments, the length of a helix is ​​8 amino acid residues. In some embodiments, the length of a helix is ​​10 amino acid residues. In some embodiments, the length of a helix is ​​12 amino acid residues. In some embodiments, the length of a helix is ​​14 amino acid residues. In some embodiments, the length of a helix is ​​16 amino acid residues. In some embodiments, the length of a helix is ​​17 amino acid residues. In some embodiments, the length of a helix is ​​18 amino acid residues. In some embodiments, the helix is ​​19 amino acid residues in length. In some embodiments, the helix is ​​20 amino acid residues in length.

[0122] The amino acids stapled together can have various numbers of amino acid residues between them, such as 1-20, 1-15, 1-10, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, the staple is (i, i+4), meaning that there are three amino acid residues (positions i+1, i+2, and i+3, respectively) between the two amino acids attached to the staple (positions i and i+4, respectively). In some embodiments, the staple is (i, i+2). In some embodiments, the staple is (i, i+3). In some embodiments, the staple is (i, i+5). In some embodiments, the staple is (i, i+6). In some embodiments, the staple is (i, i+7). In some embodiments, the staple is (i, i+8). In some embodiments, the staple peptide comprises two staples, one at (i, i+2) and the other at (i, i+7). In some embodiments, the staple peptide comprises two staples, one at (i, i+3) and the other at (i, i+7). In some embodiments, the staple peptide comprises two staples, one at (i, i+3) and the other at (i, i+4). In some embodiments, the staple peptide comprises two staples, one at (i, i+4) and the other at (i, i+7). In some embodiments, the staple peptide comprises two staples, one at (i, i+3) and the other at (i, i+3). In some embodiments, the staple peptide comprises two staples, one at (i, i+4) and the other at (i, i+4). In some embodiments, the stapled peptide comprises two staples, one at (i, i+7) and the other at (i, i+7). In some embodiments, the two staples are attached to a common backbone atom, e.g., the alpha carbon atom of an amino acid residue. In some embodiments, the stapled peptide further comprises a third staple. In some embodiments, the third staple is (i, i+3). In some embodiments, the third staple is (i, i+4). In some embodiments, the third staple is (i, i+7).In some embodiments, the staple peptide further comprises a fourth staple. In some embodiments, the fourth staple is (i, i+3). In some embodiments, the fourth staple is (i, i+4). In some embodiments, the fourth staple is (i, i+7).

[0123] In some embodiments, the stapled peptide comprises a staple, wherein the staple is L s and L s -L s1 -L s2 -L s3 - and L s1 , L s2 , and L s3 is independently L, and each L is independently as described herein. In some embodiments, a provided staple comprises L s is.

[0124] In some embodiments, L s1 contains at least one -N(R')-, where R' is as described in this disclosure. In some embodiments, -N(R')- is bonded to two carbon atoms, and neither of the two carbon atoms forms a double bond with a heteroatom. In some embodiments, -N(R')- is not bonded to -C(O)-. In some embodiments, -N(R')- is not bonded to -C(S)-. In some embodiments, -N(R')- is not bonded to -C(=NR')-. In some embodiments, L s1 is -L'-N(R')-, where L' is an optionally substituted divalent C1-C 19 In some embodiments, L s1 is -L'-N(CH3)-, where L' is an optionally substituted divalent C1-C 19 It is aliphatic.

[0125] In some embodiments, R' is optionally substituted C 1~6 In some embodiments, R' is C 1~6In some embodiments, R' is alkyl. In some embodiments, R' is methyl. s1 The peptide backbone atom to which is attached is R 1 are also bonded to R' and R 1 are both R and, together with their intervening atoms, form an optionally substituted ring as described herein. In some embodiments, the ring formed has no additional ring heteroatoms beyond the nitrogen atom to which R' is attached. In some embodiments, the ring formed is three-membered. In some embodiments, the ring formed is four-membered. In some embodiments, the ring formed is five-membered. In some embodiments, the ring formed is six-membered.

[0126] In some embodiments, L' is an optionally substituted divalent C1-C 20 In some embodiments, L' is an optionally substituted divalent C1-C 19 In some embodiments, L' is an optionally substituted divalent C1-C 15 In some embodiments, L' is an optionally substituted divalent C1-C 10In some embodiments, L' is an optionally substituted divalent C1-C9 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C8 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C7 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C6 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C5 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C4 aliphatic. In some embodiments, L' is an optionally substituted alkylene. In some embodiments, L' is an optionally substituted alkenylene. In some embodiments, L' is an unsubstituted alkylene. In some embodiments, L' is -CH2-. In some embodiments, L' is -(CH2)2-. In some embodiments, L' is -(CH2)3-. In some embodiments, L' is -(CH2)4-. In some embodiments, L' is -(CH2)5-. In some embodiments, L' is -(CH2)6-. In some embodiments, L' is -(CH2)7-. In some embodiments, L' is -(CH2)8-. In some embodiments, L' is attached to a peptide backbone atom. In some embodiments, L' is optionally substituted alkenylene. In some embodiments, L' is unsubstituted alkenylene. In some embodiments, L' is -CH2-CH=CH-CH2-.

[0127] In some embodiments, L' is optionally substituted phenylene.

[0128] In some embodiments, L s1 contains at least one —N(R′)C(O)—, where R′ is as described in this disclosure. In some embodiments, L s1 is -L'-N(R')C(O)-, where each of L' and R' is independently as described in this disclosure. s1is -L'-N(CH3)C(O)-, where L' is independently as described in this disclosure.

[0129] In some embodiments, L s1 contains at least one -C(O)O-. In some embodiments, L s1 contains at least one -C(O)O-. In some embodiments, L s1 is -L'-C(O)O- or -L'-OC(O)-, where each L' is independently as described in this disclosure. s1 is -L'-C(O)O-, where each L' is independently as described in this disclosure. s1 is -L'-OC(O)-, where each L' is independently as described in this disclosure.

[0130] In some embodiments, L s1 contains at least one -S(O)2-N(R')-, where R' is as described in this disclosure. s1 contains at least one -S(O)2-N(R')-, where R' is as described in this disclosure. s1 is -L'-N(R')-S(O)- or -L'-S(O)-N(R')-, where each of L' and R' is independently as described in this disclosure. s1 is -L'-N(R')-S(O)-, where each of L' and R' is independently as described in this disclosure. s1 is -L'-S(O)2-N(R')-, where each of L' and R' is independently as described in this disclosure. s1 is -L'-N(CH3)-S(O)2- or -L'-S(O)2-N(CH3)-, where each L' is independently as described in this disclosure. s1is -L'-N(CH3)-S(O)2-, where L' is as described herein. In some embodiments, L s1 is -L'-S(O)2-N(CH3)-, where L' is as described in this disclosure.

[0131] In some embodiments, L s1 contains at least one —O—. In some embodiments, L s1 is -L'-O-, where L' is independently as described in this disclosure.

[0132] In some embodiments, L s1 is a covalent bond.

[0133] In some embodiments, L s1 is L', where L' is as described in this disclosure.

[0134] In some embodiments, L s2 is L, where L is as described in this disclosure. In some embodiments, L s2 is L', where L' is as described herein. In some embodiments, L s2 In some embodiments, L s2 is -CH-CH=CH-CH-. In some embodiments, L s2 In some embodiments, L s2 is -(CH2)4-.

[0135] In some embodiments, L s3contains at least one -N(R')-, where R' is as described in this disclosure. In some embodiments, -N(R')- is bonded to two carbon atoms, and neither of the two carbon atoms forms a double bond with a heteroatom. In some embodiments, -N(R')- is not bonded to -C(O)-. In some embodiments, -N(R')- is not bonded to -C(S)-. In some embodiments, -N(R')- is not bonded to -C(=NR')-. In some embodiments, L s3 is -L'-N(R')-, where L' is an optionally substituted divalent C1-C 19 In some embodiments, L s3 is -L'-N(CH3)-, where L' is an optionally substituted divalent C1-C 19 It is aliphatic.

[0136] In some embodiments, L s3 contains at least one —N(R′)C(O)—, where R′ is as described in this disclosure. In some embodiments, L s3 is -L'-N(R')C(O)-, where each of L' and R' is independently as described in this disclosure. s3 is -L'-N(CH3)C(O)-, where L' is independently as described in this disclosure.

[0137] In some embodiments, L s3 contains at least one -C(O)O-. In some embodiments, L s3 contains at least one -C(O)O-. In some embodiments, L s3 is -L'-C(O)O- or -L'-OC(O)-, where each L' is independently as described in this disclosure. s3 is -L'-C(O)O-, where each L' is independently as described in this disclosure. s3is -L'-OC(O)-, where each L' is independently as described in this disclosure.

[0138] In some embodiments, L s3 contains at least one -S(O)2-N(R')-, where R' is as described in this disclosure. s3 contains at least one -S(O)2-N(R')-, where R' is as described in this disclosure. s3 is -L'-N(R')-S(O)- or -L'-S(O)-N(R')-, where each of L' and R' is independently as described in this disclosure. s3 is -L'-N(R')-S(O)-, where each of L' and R' is independently as described in this disclosure. s3 is -L'-S(O)2-N(R')-, where each of L' and R' is independently as described in this disclosure. s3 is -L'-N(CH3)-S(O)2- or -L'-S(O)2-N(CH3)-, where each L' is independently as described in this disclosure. s3 is -L'-N(CH3)-S(O)2-, where L' is as described herein. In some embodiments, L s3 is -L'-S(O)2-N(CH3)-, where L' is as described in this disclosure.

[0139] In some embodiments, L s3 contains at least one —O—. In some embodiments, L s3 is -L'-O-, where L' is independently as described in this disclosure.

[0140] In some embodiments, L s3is L', where L' is as described herein. In some embodiments, L s3 is optionally substituted alkylene. In some embodiments, L s3 is an unsubstituted alkylene.

[0141] In some embodiments, L s contains at least one -N(R')-, where R' is as described in this disclosure. In some embodiments, -N(R')- is bonded to two carbon atoms, and neither of the two carbon atoms forms a double bond with a heteroatom. In some embodiments, -N(R')- is not bonded to -C(O)-. In some embodiments, -N(R')- is not bonded to -C(S)-. In some embodiments, -N(R')- is not bonded to -C(=NR')-. In some embodiments, L s contains at least one -N(R')C(O)-, where R' is as described in this disclosure.

[0142] In some embodiments, L s , L s1 , L s2 , and L s3 each optionally independently includes an R' group, e.g., an R' group in -C(R')2-, -N(R')-, etc., and the R' group may be a group (e.g., a group that can be R) attached to a backbone atom, as can two R groups (e.g., R a1 , R a2 , R a3 , L a1 or L a2together with the R' groups in (e.g., R' groups in -C(R')2-, -N(R')-, etc.) form a double bond or an optionally substituted ring. In some embodiments, the ring formed is an optionally substituted 3- to 10-membered ring. In some embodiments, the ring formed is an optionally substituted 3-membered ring. In some embodiments, the ring formed is an optionally substituted 4-membered ring. In some embodiments, the ring formed is an optionally substituted 5-membered ring. In some embodiments, the ring formed is an optionally substituted 6-membered ring. In some embodiments, the ring formed is monocyclic. In some embodiments, the ring formed is saturated. In some embodiments, the ring formed is partially unsaturated. In some embodiments, the ring formed is aromatic. In some embodiments, the ring formed contains one or more ring heteroatoms (e.g., nitrogen). In some embodiments, the staple, or L s , L s1 , L s2 , and / or L s3 comprises -N(R')-, where R' together with the groups attached to the backbone atoms form an optionally substituted ring as described herein. s , L s1 , L s2 , and / or L s3 comprises -C(R')2-, where R' together with the groups attached to the skeletal atoms form an optionally substituted ring as described herein.

[0143] In some embodiments, staples, or L s , L s1 , L s2 , and / or L s3 includes a portion of one or more amino acid side chains (eg, a side chain other than the terminal =CH2).

[0144] As will be apparent to those of skill in the art upon reading this disclosure, the letter "L" is used to refer to a linker moiety as described herein; thus, each L 上付き文字 (For example, L a , Ls1 , L s2 , L s3 , L s ) is understood in some embodiments to be L unless otherwise specified.

[0145] In some embodiments, L comprises at least one —N(R′)—, where R′ is as described herein. In some embodiments, —N(R′)— is bonded to two carbon atoms, neither of which forms a double bond with a heteroatom. In some embodiments, —N(R′)— is not bonded to —C(O)—. In some embodiments, —N(R′)— is not bonded to —C(S)—. In some embodiments, —N(R′)— is not bonded to —C(═NR′)—. In some embodiments, L is -L′-N(R′)—, where L′ is an optionally substituted divalent C1-C 19 In some embodiments, L is -L'-N(CH)-, where L' is an optionally substituted divalent C-C 19 It is aliphatic.

[0146] In some embodiments, L comprises at least one -N(R')C(O)-, where R' is as described herein. In some embodiments, L is -L'-N(R')C(O)-, where each of L' and R' is independently as described herein. In some embodiments, L is -L'-N(CH3)C(O)-, where L' is independently as described herein.

[0147] In some embodiments, L comprises at least one -C(O)O-. In some embodiments, L comprises at least one -C(O)O-. In some embodiments, L is -L'-C(O)O- or -L'-OC(O)-, where each L' is independently as described in this disclosure. In some embodiments, L is -L'-C(O)O-, where each L' is independently as described in this disclosure. In some embodiments, L is -L'-OC(O)-, where each L' is independently as described in this disclosure.

[0148] In some embodiments, L comprises at least one -S(O)2-N(R')-, where R' is as described herein. In some embodiments, L comprises at least one -S(O)2-N(R')-, where R' is as described herein. In some embodiments, L is -L'-N(R')-S(O)2- or -L'-S(O)2-N(R')-, where each of L' and R' is independently as described herein. In some embodiments, L is -L'-N(R')-S(O)2-, where each of L' and R' is independently as described herein. In some embodiments, L is -L'-S(O)2-N(R')-, where each of L' and R' is independently as described herein. In some embodiments, L is -L'-N(CH3)-S(O)2- or -L'-S(O)2-N(CH3)-, where each L' is independently as described herein. In some embodiments, L is -L'-N(CH3)-S(O)2-, where L' is as described herein. In some embodiments, L is -L'-S(O)2-N(CH3)-, where L' is as described herein.

[0149] In some embodiments, L comprises at least one -O-. In some embodiments, L is -L'-O-, where L' is independently as described in this disclosure.

[0150] In some embodiments, L is L', where L' is as described in this disclosure. In some embodiments, L is an optionally substituted alkylene. In this embodiment, L is unsubstituted alkylene.

[0151] In some embodiments, L is an optionally substituted divalent C-C 25 In some embodiments, L is an optionally substituted divalent C1-C 20 In some embodiments, L is an optionally substituted divalent C1-C 15 In some embodiments, L is an optionally substituted divalent C1-C 10In some embodiments, L is an optionally substituted divalent C1-C9 aliphatic. In some embodiments, L is an optionally substituted divalent C1-C8 aliphatic. In some embodiments, L is an optionally substituted divalent C1-C7 aliphatic. In some embodiments, L is an optionally substituted divalent C1-C6 aliphatic. In some embodiments, L is an optionally substituted divalent C1-C5 aliphatic. In some embodiments, L is an optionally substituted divalent C1-C4 aliphatic. In some embodiments, L is an optionally substituted alkylene. In some embodiments, L is an optionally substituted alkenylene. In some embodiments, L is an unsubstituted alkylene. In some embodiments, L is -CH2-. In some embodiments, L is -(CH2)2-. In some embodiments, L is -(CH2)3-. In some embodiments, L is -(CH2)4-. In some embodiments, L is -(CH2)5-. In some embodiments, L is -(CH2)6-. In some embodiments, L is -(CH2)7-. In some embodiments, L is -(CH2)8-. In some embodiments, L is attached to a peptide backbone atom. In some embodiments, L is optionally substituted alkenylene. In some embodiments, L is unsubstituted alkenylene. In some embodiments, L is -CH2-CH=CH-CH2-.

[0152] In some embodiments, one end of the staple is at atom A of the peptide backbone. n1 is connected to A n1 is R 1 and is optionally substituted with , and is located at amino acid position n from the N-terminus of the peptide 1 The other end is the atom A of the peptide backbone. n2 is connected to A n2 is R 2 and optionally substituted with (in some embodiments, R 1 and / or R 2 is R, which may be hydrogen), and2 are atoms of amino acid residues in n 1 and n 2 are independently integers, and n 2 =n 1 +m, where m is 3 to 12.

[0153] In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5. In some embodiments, m is 6. In some embodiments, m is 7. In some embodiments, m is 8. In some embodiments, m is 9. In some embodiments, m is 10. In some embodiments, m is 11. In some embodiments, the staples are referred to as (i, i+m) staples.

[0154] In some embodiments, A n1 is a carbon atom. n1 is achiral. In some embodiments, A n1 is chiral. In some embodiments, A n1 is R. In some embodiments, A n1 is S.

[0155] In some embodiments, A n2 is a carbon atom. n2 is achiral. In some embodiments, A n2 is chiral. In some embodiments, A n2 is R. In some embodiments, A n2 is S.

[0156] In some embodiments, A n1 is achiral, and A n2 is achiral. In some embodiments, A n1 is achiral, and A n2 is R. In some embodiments, A n1 is achiral, and A n2 is S. In some embodiments, A n1is R and A n2 is achiral. In some embodiments, A n1 is R and A n2 is R. In some embodiments, A n1 is R and A n2 is S. In some embodiments, A n1 is S and A n2 is achiral. In some embodiments, A n1 is S and A n2 is R. In some embodiments, A n1 is S and A n2 is S.

[0157] In some embodiments, the provided stereochemistry at the staple backbone attachment points and / or combinations thereof, optionally in conjunction with one or more structural elements of the provided peptides, e.g., staple chemistry (hydrocarbon, non-hydrocarbon), staple length, etc., can provide various benefits, e.g., improved preparation yield, purity, and / or selectivity, improved properties (e.g., improved solubility, improved stability, reduced toxicity, improved selectivity, etc.), improved activity, etc. In some embodiments, the provided stereochemistry and / or stereochemical combinations are different from those typically used, e.g., US9617309, US2015-0225471, US2016-0024153, US2016-0215036, US2016-0244494, WO2017 / 062518, and provide one or more of the benefits described in this disclosure.

[0158] In some embodiments, staples can be of various lengths, which in some embodiments is represented by the number of staple chain atoms. In some embodiments, the staple chain is the shortest covalent connection in the staple from the first end of the staple (the point of attachment to the peptide backbone) to the second end of the staple, with the first and second ends being attached to two different peptide backbone atoms. In some embodiments, staples contain 5 to 30 chain atoms, e.g., 5 to 20, 5 to 15, 5, 6, 7, 8, 9, or 10 to 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 chain atoms. In some embodiments, staples contain 5 chain atoms. In some embodiments, staples contain 6 chain atoms. In some embodiments, staples contain 7 chain atoms. In some embodiments, staples contain 8 chain atoms. In some embodiments, staples contain 9 chain atoms. In some embodiments, the staples comprise 10 chain atoms. In some embodiments, the staples comprise 11 chain atoms. In some embodiments, the staples comprise 12 chain atoms. In some embodiments, the staples comprise 13 chain atoms. In some embodiments, the staples comprise 14 chain atoms. In some embodiments, the staples comprise 15 chain atoms. In some embodiments, the staples comprise 16 chain atoms. In some embodiments, the staples comprise 17 chain atoms. In some embodiments, the staples comprise 18 chain atoms. In some embodiments, the staples comprise 19 chain atoms. In some embodiments, the staples comprise 20 chain atoms. In some embodiments, the staples have a length of 5 chain atoms. In some embodiments, the staples have a length of 6 chain atoms. In some embodiments, the staples have a length of 7 chain atoms. In some embodiments, the staples have a length of 8 chain atoms. In some embodiments, the staples have a length of 9 chain atoms. In some embodiments, the staples have a length of 10 chain atoms. In some embodiments, the staple has a length of 11 chain atoms. In some embodiments, the staple has a length of 12 chain atoms.In some embodiments, the staples have a length of 13 chain atoms. In some embodiments, the staples have a length of 14 chain atoms. In some embodiments, the staples have a length of 15 chain atoms. In some embodiments, the staples have a length of 16 chain atoms. In some embodiments, the staples have a length of 17 chain atoms. In some embodiments, the staples have a length of 18 chain atoms. In some embodiments, the staples have a length of 19 chain atoms. In some embodiments, the staples have a length of 20 chain atoms. In some embodiments, the staples have a length of 8-15 chain atoms. In some embodiments, the staples have 8-12 chain atoms. In some embodiments, the staples have 9-12 chain atoms. In some embodiments, the staples have 9-10 chain atoms. In some embodiments, the staples have 8-10 chain atoms. In some embodiments, the length of a staple can be adjusted depending on the distance of the amino acid residues it connects, e.g., a (i, i+7) staple can be utilized that is longer than a (i, i+4) or (i, i+3) staple. In some embodiments, the (i, i+2) staple has about 5-10, 5-8, e.g., about 5, 6, 7, 8, 9, or 10 chain atoms. In some embodiments, the (i, i+2) staple has 5 chain atoms. In some embodiments, the (i, i+2) staple has 6 chain atoms. In some embodiments, the (i, i+2) staple has 7 chain atoms. In some embodiments, the (i, i+2) staple has 8 chain atoms. In some embodiments, the (i, i+2) staple has 9 chain atoms. In some embodiments, the (i, i+2) staple has 10 chain atoms. In some embodiments, the (i, i+3) staple has about 5-10, 5-8, e.g., about 5, 6, 7, 8, 9, or 10 chain atoms. In some embodiments, the (i, i+3) staple has 5 chain atoms. In some embodiments, the (i, i+3) staple has 6 chain atoms. In some embodiments, the (i, i+3) staple has 7 chain atoms. In some embodiments, the (i, i+3) staple has 8 chain atoms.In some embodiments, the (i, i+3) staple has 9 chain atoms. In some embodiments, the (i, i+3) staple has 10 chain atoms. In some embodiments, the (i, i+4) staple has about 5-12, 5-10, 7-12, 5-8, e.g., about 5, 6, 7, 8, 9, 10, 11, or 12 chain atoms. In some embodiments, the (i, i+4) staple has 5 chain atoms. In some embodiments, the (i, i+4) staple has 6 chain atoms. In some embodiments, the (i, i+4) staple has 7 chain atoms. In some embodiments, the (i, i+4) staple has 8 chain atoms. In some embodiments, the (i, i+4) staple has 9 chain atoms. In some embodiments, the (i, i+4) staple has 10 chain atoms. In some embodiments, the (i, i+4) staple has 11 chain atoms. In some embodiments, the (i, i+4) staple has 12 chain atoms. In some embodiments, the (i, i+7) staple has about 8 to 25, 10 to 25, 10 to 16, 12 to 15, e.g., about 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 chain atoms. In some embodiments, the (i, i+7) staple has 8 chain atoms. In some embodiments, the (i, i+7) staple has 9 chain atoms. In some embodiments, the (i, i+7) staple has 10 chain atoms. In some embodiments, the (i, i+7) staple has 11 chain atoms. In some embodiments, the (i, i+7) staple has 12 chain atoms. In some embodiments, the (i, i+7) staple has 13 chain atoms. In some embodiments, the (i, i+7) staple has 14 chain atoms. In some embodiments, the (i, i+7) staple has 15 chain atoms. In some embodiments, the (i, i+7) staple has 16 chain atoms. In some embodiments, the (i, i+7) staple has 17 chain atoms. In some embodiments, the (i, i+7) staple has 18 chain atoms. In some embodiments, the (i, i+7) staple has 19 chain atoms.In some embodiments, the (i, i+7) staple has 20 chain atoms. In some embodiments, the (i, i+7) staple has 21 chain atoms. In some embodiments, the (i, i+7) staple has 22 chain atoms. In some embodiments, the staple peptide comprises three or more staples, each of which is independently such a (i, i+2), (i, i+3), (i, i+4), or (i, i+7) staple. In some embodiments, the staple peptide comprises such a (i, i+2) staple, such a (i, i+4) staple, and such a (i, i+7) staple. In some embodiments, the staple peptide comprises such a (i, i+3) staple, such a (i, i+4) staple, and such a (i, i+7) staple. In some embodiments, the staple peptides include such an (i, i+3) staple, such an (i, i+7) staple, and such an (i, i+7) staple.

[0159] Staple length can be described in other ways. For example, in some embodiments, staple length can be described as the total number of chain atoms and non-chain ring atoms, where a non-chain ring atom is an atom of the staple that forms a ring with one or more chain atoms, but is not a chain atom in that it is not within the shortest covalent connection from the first end of the staple to the second end of the staple. In some embodiments, staples formed using monomer A (containing an azetidine moiety), monomer B (containing a pyrrolidine moiety), and / or monomer C (containing a pyrrolidine moiety), etc., contain one or two non-chain ring atoms. obtain.

[0160] In some embodiments, the staple has no heteroatoms in the chain. In some embodiments, the staple contains at least one heteroatom in the chain. In some embodiments, the staple contains at least one nitrogen atom in the chain.

[0161] In some embodiments, the staple is L swhere L s is an optionally substituted divalent C 8~14 is an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, the staple is s where L s is an optionally substituted divalent C 9~13 is an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, the staple is s where L s is an optionally substituted divalent C 10~15 is an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, the staple is s where L s is an optionally substituted divalent C 11~14an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, the staple is an (i, i+2) staple, in that it does not include the two amino acid residues directly attached to the staple, but rather there is one amino acid residue between the two amino acid residues directly attached to the staple. In some embodiments, the staple is an (i,i+3) staple in that it does not include two amino acid residues directly attached to the staple, and there are two amino acid residues between the two amino acid residues directly attached to the staple. In some embodiments, the staple is an (i,i+4) staple in that it does not include two amino acid residues directly attached to the staple, and there are three amino acid residues between the two amino acid residues directly attached to the staple. In some embodiments, the staple is an (i,i+7) staple in that it does not include two amino acid residues directly attached to the staple, and there are six amino acid residues between the two amino acid residues directly attached to the staple.

[0162] In some embodiments, L s , L s1 , L s2 , and L s3 For each of L, any replacement of a methylene unit, if present, is with -N(R')-, -C(O)-N(R')-, -N(R')C(O)O-, -C(O)O-, -S(O)N(R')-, or -O-. In some embodiments, L s , L s1 , L s2 , and L s3 For each of L, any replacement of a methylene unit, if present, is with -N(R')-, -N(R')-C(O)-, or -N(R')C(O)O-. s , Ls1 , L s2 , and L s3 For each of L, any replacement of a methylene unit, if present, is with -N(R')- or -N(R')C(O)O-. s , L s1 , L s2 , and L s3 For each of the methylene units, if any replacement is present, it is with -N(R')-. s , L s1 , L s2 , and L s3 For each of the groups, if any replacement of a methylene unit is present, it is by -N(R')C(O)O-.

[0163] In some embodiments, the staple comprises a double bond. In some embodiments, the staple comprises a double bond that can be formed by olefin metathesis of two olefins. In some embodiments, the staple is formed by a metathesis reaction involving one or more double bonds in an amino acid residue, for example, as described herein. In some embodiments, a first amino acid residue comprising an olefin (e.g., AA1-CH=CH2) and a second amino acid residue comprising an olefin (e.g., AA2-CH=CH2) are stapled (e.g., AA1-CH=CH-AA2 is formed, where AA1 and AA2 are typically linked through one or more amino acid residues). In some embodiments, for example, an olefin in a staple is converted to -CHR'-CHR'-, where each R' is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R' is -H. In some embodiments, each R' is -H. In some embodiments, R' is -OR, where R is as described herein. In some embodiments, R' is -OH. In some embodiments, R' is -N(R)2, where each R is independently as described herein. In some embodiments, R' is -SR, where R is as described herein. In some embodiments, R' is R, where R is an optionally substituted aliphatic, e.g., C 1~10 In some embodiments, R' is R, where R is an optionally substituted aliphatic, such as C 1~10 In some embodiments, R' is R, where R is an optionally substituted aliphatic, such as C 1~10 In some embodiments, -CHR'-CHR'- is -CH2-CH2-. In some embodiments, each of the two olefins is independently a side chain of an amino acid residue. In some embodiments, each olefin is independently a terminal olefin. In some embodiments, each olefin is independently a monosubstituted olefin.

[0164] In some embodiments, the amino acid of formula AI or salt thereof has the structure of formula A-II. NH(R a1 )-L a1 -C(-L a -CH=CH2)(R a3 )-L a2 -COOH A-II or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of formula A-II or a salt thereof, wherein each variable is independently as described in this disclosure.

[0165] In some embodiments, the amino acid of formula A-II or salt thereof has the structure of formula A-II-b NH(R a1 )-C(-L a -CH=CH2)(R a3 )-COOH A-II-b or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of formula A-II-b or a salt thereof, wherein each variable is independently as described in this disclosure.

[0166] In some embodiments, the amino acid of formula AI or salt thereof has the structure of formula A-III. N(-L a -CH=CH2)(R a1 )-L a1 -C(-L a -CH=CH2)(R a3 )-L a2 -COOH A-III or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of formula A-II or a salt thereof, wherein each variable is independently as described in this disclosure.

[0167] In some embodiments, the amino acid of formula AI or salt thereof is represented by formula A-IV: NH(R a1 )-L a1 -C(-L a -COOH)(R a3 )-L a2 -COOH, A-IV or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of Formula A-IV, or a salt thereof, wherein each variable is independently as described in this disclosure.

[0168] In some embodiments, the amino acid has the formula AV: NH(R a1 )-L a1 -C(-L a -R SP1 )(R a3 )-L a2 -COOH, AV or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of formula AV or a salt thereof, wherein each variable is independently as described in this disclosure.

[0169] In some embodiments, the amino acids for stapling are represented by Formula A-VI: NH(R a1 )-L a1 -C(-L a -R SP1 )(-L a -R SP2 )-L a2 -COOH, A-VI or a salt thereof, wherein each variable is independently as described in this disclosure. In some embodiments, an amino acid suitable for stapling has the structure of Formula A-VI or a salt thereof, wherein each variable is independently as described in this disclosure.

[0170] As used herein, R SP1 and R SP2 Each of R independently comprises a reactive group. SP1 and R SP2 are independently a reactive group. In some embodiments, the reactive group is an optionally substituted -CH=CH2. In some embodiments, the reactive group is -CH=CH2. In some embodiments, the reactive group is an amino group, e.g., -NHR, where R is as described herein. In some embodiments, the reactive group is an acid group. In some embodiments, the reactive group is -COOH or an activated form thereof. In some embodiments, the reactive group is for cycloaddition reactions (e.g., [3+2], [4+2], etc.), e.g., an alkene, alkyne, diene, 1,3-dipole (e.g., -N3), etc. In some embodiments, the reactive group is an optionally substituted -C≡CH. In some embodiments, the reactive group is -C≡CH. In some embodiments, the reactive group is -N3.

[0171] In some embodiments, the R of the first amino acid residue SP1 or R SP2 and R of the second amino acid residue SP1 or R SP2can react with each other, thus connecting two amino acid residues with a staple. In some embodiments, the reaction is olefin metathesis between two olefins, e.g., two -CH=CH2. In some embodiments, the reaction is amidation, where one reactive group, is an amino group, e.g., -NHR, where R is as described herein (e.g., in some embodiments, R is -H; in some embodiments, R is an optionally substituted C 1~6 In some embodiments, one reactive group is or comprises -N3, and the other reactive group is or comprises an alkyne, e.g., a terminal alkyne or an activated / strained alkyne. In some embodiments, the other reactive group is or comprises -C≡CH.

[0172] In some embodiments, the R of the first amino acid residue SP1 or R SP2 and R of the second amino acid residue SP1 or R SP2 can react with the reagent, thus connecting the two amino acid residues to form a staple. In some embodiments, the reagent contains two reactive groups, one at R of the first amino acid residue. SP1 or R SP2 and the other reacts with R of the first amino acid residue. SP1 or R SP2 In some embodiments, the R SP1 or R SP2 are the same or of the same type, e.g., both are amino groups, and the two reactive groups of the linking reagent are also the same, e.g., both are acid groups such as —COOH or an activated form thereof. In some embodiments, the RSP1 or R SP2 are both acid groups, e.g., —COOH or activated forms thereof, and both reactive groups of the linker are amino groups. In some embodiments, the R SP1 or R SP2 are both nucleophilic groups, e.g., -SH, and both reactive groups of the linking reagent are electrophilic (e.g., carbons attached to leaving groups such as -Br, -I, etc.).

[0173] In some embodiments, R SP1 and R SP2 In some embodiments, R SP1 and R SP2 In some embodiments, R SP1 is or includes -CH=CH2. In some embodiments, R SP1 is or includes -COOH. In some embodiments, R SP1 is or includes an amino group. In some embodiments, R SP1 In some embodiments, R is or includes -NHR. In some embodiments, R is hydrogen or optionally substituted C 1~6 In some embodiments, R SP1 is or includes -NH. In some embodiments, R SP1 is or includes -N3. In some embodiments, R SP2 is or includes -CH=CH2. In some embodiments, R SP2 is or includes -COOH. In some embodiments, R SP2 is or includes an amino group. In some embodiments, R SP2 In some embodiments, R is or includes -NHR. In some embodiments, R is hydrogen or optionally substituted C 1~6 In some embodiments, R SP2 is or includes -NH. In some embodiments, R SP2 is or includes -N3.

[0174] In some embodiments, each amino acid residue of a pair of amino acid residues is independently a residue of an amino acid of formula A-II or A-III, or a salt thereof. In some embodiments, such pairs of amino acid residues are stapled together, for example, by olefin metathesis. In some embodiments, the staple is -L a -CH=CH-L a wherein each variable is independently as described herein. In some embodiments, the olefin in the staple is reduced. In some embodiments, the staple has the structure -L a -CH2-CH2-L a -, wherein each variable is independently as described herein. In some embodiments, one L a is the L described herein s1 and one L a is the L described herein s3 is.

[0175] In some embodiments, two amino acid residues of the amino acid of formula AI or a salt thereof connected by a staple, e.g., independently, are joined by a staple, and each amino acid residue is selected from the group consisting of -N(R a1 )-L a1 -C(-L s -R AA )(R a3 )-L a2 -CO-, where each variable is independently as described herein; AA is an amino acid residue. In some embodiments, two amino acid residues of the amino acid of formula AI or a salt thereof connected by a staple, e.g., independently, are joined by a staple. s -R AA )-L a1 -C(R a2 )(R a3 )-L a2 -CO-, where each variable is independently as described herein; AAis an amino acid residue. In some embodiments, two amino acid residues connected by a staple, e.g., independently, an amino acid of formula AI or a salt thereof, are joined by R a1 -N(-L s -R AA )-L a1 -C(R a2 )(R a3 )-L a2 -CO-, where each variable is independently as described herein; AA is an amino acid residue. In some embodiments, two stapled, e.g., three amino acid residues of an amino acid of formula AI or a salt thereof are joined by R a1 -N(-L s -R AA )-L a1 -C(-L s -R AA )(R a3 )-L a2 -CO-, where each variable is independently as described herein; AA is an amino acid residue. In some embodiments, two stapled, e.g., three amino acid residues of an amino acid of formula AI or a salt thereof are independently -N(-L s -R AA )-L a1 -C(-L s -R AA )(R a3 )-L a2 -CO-, where each variable is independently as described herein; AA is an amino acid residue. In some embodiments, two stapled, e.g., three amino acid residues of an amino acid of formula AI or a salt thereof (e.g., X 1 and X 14 Both the and stapled X 4 ) is -N(R a1 )-L a1 -C(-L s -R AA )(-L s -R AA )-L a2-CO-, where each variable is independently as described herein; AA is an amino acid residue. In some embodiments, each R AA are independently a residue of an amino acid or salt thereof of formula AI, A-II, A-III, A-IV, AV, A-VI, etc. In some embodiments, R AA is -C(R a3 )[-L a1 -N(R a1 )-](-L a2 -CO-), where each variable is independently as described herein. In some embodiments, R AA is -C(R a3 )[-N(R a1 )-](-CO-), wherein each variable is independently as described herein. In some embodiments, each R AA are independently -N(-)[-L a1 -C(R a2 )(R a3 )-L a2 -CO-], wherein each variable is independently as described herein, and wherein -C(-)(R a3 )- is attached to the staple. In some embodiments, each R AA are independently -N(-)[-C(R a2 )(R a3 )—CO—], wherein each variable is independently as described herein, and wherein —C(—)(R a3 )- is attached to the staple. In some embodiments, each R AA are independently a1 -N(-)[-L a1 -C(R a2 )(R a3 )-L a2 -CO-], wherein each variable is independently as described herein, and wherein -C(-)(R a3 )- is attached to the staple. In some embodiments, each R AA are independently a1 -N(-)[-C(Ra2 )(R a3 )—CO—], wherein each variable is independently as described herein, and wherein —C(—)(R a3 )- is attached to the staple.

[0176] Various staples, e.g., L s is as described herein. In some embodiments, L s is -L as described herein s1 -L s2 -L s3 In some embodiments, L s1 is the L described herein a In some embodiments, L s3 is the L described herein a In some embodiments, L s1 is the first of the two stapled amino acid residues a In some embodiments, L s2 is the second of the two stapled amino acid residues a In some embodiments, L s2 is or contains a double bond. s2 is or includes -CH=CH-. In some embodiments, L s2 is or includes optionally substituted -CH-CH-. In some embodiments, L s2 is or includes -CH-CH-. In some embodiments, L s2 is -C(O)N(R')- (e.g., R SP1 R having a group, and the other is or contains -COOH SP2 In some embodiments, L is or comprises a staple formed by two amino acid residues having a hydroxyl group. s2 is or includes —C(O)NH—. In some embodiments, L s1 and L s3 each independently being an optionally substituted straight or branched chain C1~10 In some embodiments, L s1 and L s3 Each of is independently —(CH)—, where n is 1 to 10. In some embodiments, L s1 is -CH-. In some embodiments, L s3 is -(CH2)3-.

[0177] In some embodiments, L s is -CH-CH=CH-(CH)-. In some embodiments, L s is -(CH2)6-.

[0178] In some embodiments, L s is —(CH2)2—C(O)NH—(CH2)4—.

[0179] In some embodiments, L s is attached to two backbone carbon atoms. s are attached to the two alpha carbon atoms of the two stapled amino acid residues. s is attached to a backbone nitrogen atom and a backbone carbon atom (eg, the alpha carbon).

[0180] In some embodiments, L a contains at least one -N(R')-, where R' is independently as described in this disclosure. a -L am1 -N(R')-, where R' is independently as described in this disclosure; am1 is as described herein. In some embodiments, L a -L am1 -N(R')-L am2 - or containing -, wherein L am1 , R', and L am2 and R' is independently as described herein. In some embodiments, R' is optionally substituted C 1~6In some embodiments, R' is methyl. In some embodiments, R' is R a3 and together form an optionally substituted ring as described herein. In some embodiments, the ring formed is a 3- to 10-membered monocyclic saturated ring as described herein. In some embodiments, the ring formed has no additional heteroatom ring atoms beyond the nitrogen of -N(R')-. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6-membered.

[0181] In some embodiments, L a contains at least one -C(R')2-, where each R' is independently as described in this disclosure. a -L am1 -C(R')2-, where R' is independently as described in this disclosure; am1 is as described herein. In some embodiments, L a -L am1 -C(R')2-L am2 - or containing L am1 , R', and L am2 are independently as described herein. In some embodiments, R' is -H. In some embodiments, -C(R')2- is optionally substituted -CH2-. In some embodiments, -C(R')2- is -CH2-. In some embodiments, one R' is R a3 and together form an optionally substituted ring as described herein. In some embodiments, the ring formed is a 3- to 10-membered monocyclic saturated ring as described herein. In some embodiments, the ring formed has no additional heteroatom ring atoms beyond the nitrogen of -N(R')-. In some embodiments, the ring formed is 3-membered. In some embodiments, the ring formed is 4-membered. In some embodiments, the ring formed is 5-membered. In some embodiments, the ring formed is 6-membered.

[0182] As described herein, L am1 and L am2 Each of the groups independently represents a group selected from the group consisting of L, am As described herein, L am is a covalent bond or an optionally substituted divalent C1-C 10 is an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L am is a covalent bond. In some embodiments, L am is an optionally substituted divalent C1-C 10 In some embodiments, L is an aliphatic group. am is an optionally substituted divalent, linear C1-C 10 In some embodiments, L is an aliphatic group. am is replaced by C 1~10 In some embodiments, L is alkylene. am is C 1~10 In some embodiments, L is alkylene. am is an optionally substituted linear C 1~10 In some embodiments, L is alkylene. am is optionally substituted —CH—. In some embodiments, L am is -CH2-.

[0183] In some embodiments, L am1 is a covalent bond. In some embodiments, L am1 is an optionally substituted divalent C1-C 10 In some embodiments, L is an aliphatic group. am1 is an optionally substituted divalent, linear C1-C 10In some embodiments, L is an aliphatic group. am1 is replaced by C 1~10 In some embodiments, L is alkylene. am1 is C 1~10 In some embodiments, L is alkylene. am1 is an optionally substituted linear C 1~10 In some embodiments, L is alkylene. am1 is optionally substituted —CH—. In some embodiments, L am1 is -CH-. In some embodiments, L am1 is attached to a backbone atom. am1 is attached to the alpha-carbon of an amino acid.

[0184] In some embodiments, L am2 is a covalent bond. In some embodiments, L am2 is an optionally substituted divalent C1-C 10 In some embodiments, L is an aliphatic group. am2 is an optionally substituted divalent, linear C1-C 10 In some embodiments, L is an aliphatic group. am2 is replaced by C 1~10 In some embodiments, L is alkylene. am2 is C 1~10 In some embodiments, L is alkylene. am2 is an optionally substituted linear C 1~10 In some embodiments, L is alkylene. am2 is optionally substituted —CH—. In some embodiments, L am2 is -CH-. In some embodiments, L am2 is or includes -C(O)-. In some embodiments, -C(O)- is attached to a nitrogen atom. In some embodiments, L am2 is or includes -S(O)-. In some embodiments, -S(O)- is attached to a nitrogen atom. In some embodiments, L am2In some embodiments, L am2 is or includes -C(O)-O-. In some embodiments, -C(O)-O- is attached to a nitrogen atom. In some embodiments, L am2 is attached to a nitrogen atom and includes a —C(O)— group attached to the nitrogen atom. am2 is attached to a nitrogen atom and includes a —C(O)—O— group attached to the nitrogen atom. am2 is or includes -C(O)-O-CH-, where -CH- is optionally substituted. In some embodiments, L am2 is -C(O)-O-CH2-.

[0185] In some embodiments, L a is the L described herein s1 In some embodiments, L a is the L described herein s2 is.

[0186] In some embodiments, R a3 -L a -CH=CH2, where L a are independently as described herein. In some embodiments, R a2 and R a3 Each of R independently contains a double bond, e.g., a terminal olefin, which may optionally be independently stapled to another residue containing an olefin. a2 and R a3 Each of the -L a In some embodiments, the amino acid is a group consisting of two amino acid residues and, independently, R a2 and R a3 In some embodiments, such an amino acid is B5. In some embodiments, it is B3. In some embodiments, it is B4. In some embodiments, it is B6.

[0187] In some embodiments, the amino acid is selected from Tables AI, A-II, A-III, and A-IV (which may be present as Fmoc-protected). As will be appreciated by those skilled in the art, Fmoc-protected amino and carboxyl groups, among others, when incorporated into a peptide, may independently form amide bonds with other amino acid residues (or may exist as N- or C-terminal capping groups, or N- or C-terminal amino or carboxyl groups). Olefins, including those within Alloc groups, can be utilized to form staples by olefin metathesis. Olefin-containing staples can be further modified, for example, by hydrogenation to convert the olefin double bond to a single bond and / or by CO extrusion to convert a carbamate moiety (e.g., -O-(CO)-N(R')-) to an amine moiety (e.g., -N(R')-). In some embodiments, the agent is or comprises a stapled peptide (e.g., a stapled peptide described according to Table E2 or Table E3), or a salt thereof, in which each double bond is converted to a single bond. In some embodiments, the conversion is accomplished by hydrogenation, whereby -H is added to each olefinic carbon atom. In some embodiments, the olefinic double bond is replaced with -CHR'-CHR'-, where each R' is independently as described herein. In some embodiments, R' is R, as described herein. In some embodiments, R' is -H. In some embodiments, each R' is -H. In some embodiments, R' is -OR, where R is as described herein. In some embodiments, R' is -OH. In some embodiments, R' is -N(R)2, where each R is independently as described herein. In some embodiments, R' is -SR, where R is as described herein. In some embodiments, R' is R, where R is an optionally substituted aliphatic group, e.g., C 1~10 In some embodiments, R' is R, where R is an optionally substituted aliphatic, such as C 1~10In some embodiments, R' is R, where R is an optionally substituted aliphatic, such as C 1~10 In some embodiments, -CHR'-CHR'- is -CH2-CH2-.

[0188] Table AI. Exemplary amino acids (Fmoc-protected). [Table 2]

[0189] Table A-II. Exemplary amino acids (Fmoc-protected). [Table 3]

[0190] Table A-III. Exemplary amino acids (Fmoc-protected). [Table 4-1] [Table 4-2]

[0191] In some embodiments, the amino acid is an alpha-amino acid. In some embodiments, the amino acid is an L-amino acid. In some embodiments, the amino acid is a D-amino acid. In some embodiments, the alpha-carbon of the amino acid is achiral. In some embodiments, the amino acid is a beta-amino acid. In some embodiments, the amino acid is a gamma-amino acid.

[0192] In some embodiments, the provided amino acid sequences contain two or more amino acid residues whose side chains link to form one or more staples. In some embodiments, the provided amino acid sequences contain two or more amino acid residues, each of which independently has a side chain comprising an olefin. In some embodiments, the provided amino acid sequences contain two or more amino acid residues, each of which independently has a side chain comprising a terminal olefin. In some embodiments, the provided amino acid sequences contain no more than two amino acid residues, each of which independently has a side chain comprising an olefin. In some embodiments, the provided amino acid sequences contain no more than two amino acid residues, each of which independently has a side chain comprising a terminal olefin. In some embodiments, the provided amino acid sequences contain at least one amino acid residue comprising an olefin and a nitrogen atom other than the nitrogen atom of the amino group. In some embodiments, the provided amino acid sequences contain at least one amino acid residue comprising a terminal olefin and a nitrogen atom other than the nitrogen atom of the amino group. In some embodiments, the provided amino acid sequences contain at least one amino acid residue having a side chain comprising a terminal olefin and a nitrogen atom. In some embodiments, the provided amino acid sequence comprises a residue of at least one amino acid of formula AI, where R a2 comprises an olefin and an —N(R′)— moiety, where R′ is as described herein (in some embodiments, optionally R a3 and together with their intervening atoms form an optionally substituted ring as described herein). In some embodiments, R a2 comprises a terminal olefin and an -N(R')- moiety, where R' is as described in this disclosure. In some embodiments, the provided amino acid sequences comprise a residue of at least one amino acid selected from Table AI. In some embodiments, the provided amino acid sequences comprise a residue of at least one amino acid selected from Table A-II. In some embodiments, the provided amino acid sequences comprise a residue of at least one amino acid selected from Table A-III. In some embodiments, the staple is formed by linking two olefins from two side chains via olefin metathesis to form a staple. In some embodiments, the staple is preferably formed by the side chains of amino acid residues that are not present at the corresponding positions of the target of interest. In some embodiments, the formed staple does not interfere with the interaction of the peptide with the target of interest.

[0193] In some embodiments, the provided staples are hydrocarbon staples. In some embodiments, the hydrocarbon staples do not contain chain heteroatoms, and the staple chain is the shortest covalent connection within the staple, from one end of the staple to the other end of the staple.

[0194] In some embodiments, the olefin in the staple is a Z-olefin. In some embodiments, the olefin in the staple is an E-olefin. In some embodiments, provided compositions include staple peptides comprising staples containing Z-olefins and staple peptides comprising staples containing E-olefins. In some embodiments, provided compositions include staple peptides comprising staples containing Z-olefins. In some embodiments, provided compositions include staple peptides comprising staples containing E-olefins. In some embodiments, otherwise identical staple peptides that differ only in the E / Z configuration of the staple olefin exhibit different properties and / or activities, as demonstrated herein. In some embodiments, given the circumstances, a staple peptide having an E-olefin in the staple may provide certain desirable properties and / or activities. In some embodiments, given the circumstances, a staple peptide having a Z-olefin in the staple may provide certain desirable properties and / or activities.

[0195] In some embodiments, the present disclosure provides a composition comprising a staple peptide. In some embodiments, the composition comprises only one stereoisomer of the staple peptide (e.g., an E or Z isomer, and / or a single diastereomer / enantiomer about a chiral center, etc.). In some embodiments, the composition comprises two or more stereoisomers (e.g., both E and Z isomers of one or more double bonds, and / or one or more diastereomers / enantiomers about a chiral center, etc.). In some embodiments, the composition corresponds to a single peak in a separation by chromatography, for example, HPLC. In some embodiments, the peak comprises only one stereoisomer. In some embodiments, the peak comprises two or more stereoisomers.

[0196] In some embodiments, two staples may be attached to the same atom of the peptide backbone, thereby forming a stitched peptide.

[0197] In some embodiments, the staple is a pro-lock, where one end of the staple is attached to the alpha-carbon of a proline residue.

[0198] In some embodiments, the staple is a staple exemplified in Tables S-1, S-2, S-3, S-4, and S-5 below (exemplary peptide backbones are illustrated for clarity (can be applied to other peptide backbones), and each X is independently an amino acid residue). In some embodiments, the staple is a staple in Table S-6 (amino acid residues attached to the staple are illustrated). In some embodiments, the olefin is Z. In some embodiments, the olefin is E. In some embodiments, the (i, i+3) staple is selected from Table S-1. In some embodiments, the (i, i+3) staple is selected from Table S-2. When a staple from Tables S-1 and S-2 is utilized for (i, i+3), "X3" in these tables is replaced with "X2" (i.e., , two amino acid residues instead of three amino acid residues). In some embodiments, the (i, i+4) staples are selected from Table S-1. In some embodiments, the (i, i+4) staples are selected from Table S-2. In some embodiments, the (i, i+7) staples are selected from Table S-3. In some embodiments, the (i, i+7) staples are selected from Table S-4.

[0199] Table S-1. Exemplary staples. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4]

[0200] Table S-2. Exemplary staples. [Table 6-1] [Table 6-2] [Table 6-3]

[0201] Table S-3. Exemplary staples. [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6] [Table 7-7]

[0202] Table S-4. Exemplary staples. [Table 8-1] [Table 8-2]

[0203] Certain useful staples are described, for example, in WO2019 / 051327, WO2022 / 020652, etc., which are incorporated herein by reference.

[0204] In some embodiments, the staple can be one of the following, connected to the amino acid at the position shown: Table S-5. Certain amino acids and staples. [Table 9-1] [Table 9-2] [Table 9-3] [Table 9-4] [Table 9-5] [Table 9-6] Table 9-7 Table 9-8 Table 9-9 Table 9-10 Table 9-11 Table 9-12 Table 9-13 Table 9-14

[0205] In some embodiments, the peptide comprises a staple or stitch (two staples) from Table S-6. In Table 6, the amino acid residues can be either N to C or C to N. In some embodiments, the amino acid residues are N to C. In some embodiments, the amino acid residues are C to N. In some embodiments, the double bond is E. In some embodiments, the double bond is Z. In some embodiments, the staple is an (i, i+2) staple. In some embodiments, the staple is an (i, i+3) staple. In some embodiments, the staple is an (i, i+4) staple. In some embodiments, the staple is an (i, i+7) staple. In some embodiments, when the structure comprises more than one double bond, each double bond is independently E or Z. In some embodiments, each staple is independently an (i, i+2) or (i, i+3) or (i, i+4) staple or (i, i+7) staple. In some embodiments, each staple is independently an (i, i+2) or (i, i+4) staple or (i, i+7) staple. In some embodiments, each staple is independently an (i, i+3) or (i, i+4) staple or (i, i+7) staple. In some embodiments, in a structure comprising two staples, each staple is independently an (i, i+4) staple or an (i, i+7) staple. In some embodiments, one staple is an (i, i+4) staple and the other staple is an (i, i+7) staple. In some embodiments, one staple is an (i, i+3) staple, one staple is an (i, i+4) staple, and one staple is an (i, i+7) staple. In some embodiments, one staple is the (i, i+2) staple, one staple is the (i, i+4) staple, and one staple is the (i, i+7) staple. In some embodiments, the PL3 residue is attached to the (i, i+3) staple. In some embodiments, the PL3 residue is attached to the (i, i+4) staple. In some embodiments, the staple (e.g.,(those in Table 6) are metathesis of double bonds in the side chains of amino acid residues, e.g., RdN and S7, R8 and PyrS, R5 and SeN, R6 and SeN, ReN and S5, ReN and S6, R7 and PyrS, Az and S7, R8 and SgN, Az and S8, R4 and SeN, R5 and SdN, R7 and Az, R8 and Az, RdN and S4, RgN and S8, RgN and S7, R8 and S5, PL3 and B5 and the same B5 and S8, PL3 and B5 and the same B5 and SeN, PL3 and B5 and the same B5 and SdN, PL3 and B5 and the same B5 and S7, PL3 and B5 and the same B5 and Pyr S2, PL3 and B5 and the same B5 and PyrS3, R5 and PyrS2, PL3 and B5 and the same B5 and PyrS1, PL3 and B5 and the same B5 and S10, PL3 and B5 and the same B5 and PyrR2, ​​PL3 and B5 and the same B5 and PyrS, PL3 and B5 and the same B5 and Az, PL3 and B5 and the same B5 and SeNc5, HypEs5 and B5 and the same B5 and PyrS2, HypEs4 and B5 and the same B5 and PyrS2, ProSAm3 and B5 and the same B5 and PyrS2, ProAm5 and B5 and the same B5 and PyrS2, ProAm6 and B5 and the same B5 and PyrS2, BzAm3O allyl and B5 and the same B5 and PyrS2, HypBzEs3O allyl and B5 and the same B5 and PyrS2, ProBzAm3O allyl and B5 and the same B5 and PyrS2, PAc3O allyl and B5 and the same B5 and PyrS2, ProPAc3O allyl and B5 and the same B5 and PyrS2, HypPAc3O allyl and B5 and the same B5 and PyrS2, Bn3O allyl and B5 and the same B5 and PyrS2, R3 and B5 and the same B5 and PyrS2, R5 and B5 and the same B5 and PyrS2, [Bz Am2 allele] MePro and B5 and the same B5 and PyrS2, PL3 and B5 and the same B5 and SPip1, PL3 and B5 and the same B5 and SPip2, PL3 and B5 and the same B5 and SPip3, PL3 and B5 and the same B5 and Az2, PL3 and B5 and the same B5 and Az3, PL3 and S5, R5 and S5, PL3 and B4 and the same B4 and PyrS1, PL3 and B4 and the same B4 and PyrS2, PL3 and B4 and the same B4 and PyrS3, PL3 and S6, PL3 and S4, PL3 and S3, R6 and PyrS2, R4 and PyrS2, R3 and PyrS2,Formed by metathesis of PL3 and B3 and the same B3 and PyrS2, PL3 and B3 and the same B3 and PyrS3, PL3 and B3 and the same B3 and PyrS4, PL3 and B6 and the same B6 and PyrS, PL3 and B6 and the same B6 and PyrS1, PL3 and B6 and the same B6 and PyrS2. Table S-6. Certain staples (including the amino acid residues that bind to the staples). [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5] [Table 10-6] [Table 10-7] [Table 10-8] [Table 10-9] [Table 10-10] [Table 10-11] [Table 10-12] [Table 10-13]

[0206] In some embodiments, the double bond of the (i, i+3) staple is Z. In some embodiments, the double bond of the (i, i+4) staple is Z. In some embodiments, the double bond of the (i, i+7) staple is Z. In some embodiments, the double bond of the (i, i+3) staple is E. In some embodiments, the double bond of the (i, i+4) staple is E. In some embodiments, the double bond of the (i, i+7) staple is E.

[0207] In some embodiments, the staple comprises -S-. In some embodiments, the stapling technique comprises the use of one or more, for example, two or more, sulfur-containing moieties. In some embodiments, the stapled peptide comprises cysteine ​​stapling. In some embodiments, two cysteine ​​residues are stapled, where the -S- moieties of the two cysteine ​​residues are optionally connected by a linker. In some embodiments, the stapled peptide comprises one from cysteine ​​stapling and one or less staples. In some embodiments, the stapled peptide comprises [ka] In some embodiments, the staple peptide comprises one or less staples having the structure: [ka] In some embodiments, the staple peptide comprises one or less staples having the structure: [ka] In some embodiments, the staple peptide comprises one or less staples having the structure: [ka] In some embodiments, the staple peptide comprises one or less staples having the structure: [ka] In some embodiments, the staple peptide does not include a staple having the structure: [ka] In some embodiments, the staple peptide does not include a staple having the structure: [ka] In some embodiments, the staple peptide does not include a staple having the structure: [ka] This does not include staples having the following structure.

[0208] In some embodiments, the present disclosure provides useful techniques related to cysteine ​​stapling. Among other things, the present disclosure recognizes that peptides suitable for cysteine ​​stapling and / or peptides containing one or more cysteine ​​staples can be produced and / or evaluated in biological systems. The present disclosure further recognizes that certain such systems enable the development, production, and / or evaluation of cysteine-stapled peptides having a range of different structures (e.g., different amino acid sequences), and may, in fact, provide the user with complete control over the selection and implementation of the amino acid sequence to be incorporated into the stapled peptide.

[0209] Cysteine ​​stapling, as described herein, involves linking one cysteine ​​residue to another, where the resulting bond is not through the peptide backbone between the cysteine ​​residues involved.

[0210] In some embodiments, the stapled peptides described herein include L s a staple comprising: L s -L s1 -SL s2 -SL s3 - and L s1 and L s3 are each independently L; L s2 is L and contains at least one -C(O)-; Each L is independently a covalent bond or an optionally substituted divalent C1-C6 25 an aliphatic group, wherein one or more methylene units of the aliphatic group are optionally independently replaced by —C(R′)2—, —Cy-, —O—, —S—, —SS-, —N(R′)—, —C(O)—, —C(S)—, —C(NR′)—, —C(O)N(R′)—, —N(R′)C(O)N(R′)—, —N(R′)C(O)O—, —S(O)—, —S(O)2N(R′)—, —C(O)S—, or —C(O)O—; Each -Cy- is independently C 3~20 Alicyclic ring, C 6~20 an optionally substituted divalent group selected from an aryl ring, a 5-20 membered heteroaryl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and a 3-20 membered heterocyclyl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon; each R' is independently -R, -C(O)R, -COR, or -SOR; Each R is independently -H or C 1~30 C having 1 to 10 heteroatoms independently selected from aliphatic, oxygen, nitrogen, sulfur, phosphorus, and silicon 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 C having 1 to 10 heteroatoms independently selected from arylaliphatic, oxygen, nitrogen, sulfur, phosphorus, and silicon 6~30an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon; or two R groups, optionally independently, taken together to form a covalent bond, or two or more R groups on the same atom optionally independently combine with the atom to form an optionally substituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having, in addition to the atom, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon; or Two or more R groups on two or more atoms optionally independently combine with their intervening atoms to form an optionally substituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon.

[0211] In some embodiments, L is independently a divalent C1-C 25 In some embodiments, L is independently a divalent C1-C6 20 In some embodiments, L is independently a divalent C1-C6 10 In some embodiments, L is independently a divalent C1-C5 aliphatic group. In some embodiments, L is independently a divalent C1 aliphatic group. In some embodiments, L is -CH2.

[0212] In some embodiments, L s1 is -CH-. In some embodiments, L s3 is -CH-. In some embodiments, L s1 and L s3 In some embodiments, L s is -CH2-SL s2-S-CH2-.

[0213] In some embodiments, L s2 is -C(R')2-L'-C(R')2-, where L' is described in this disclosure. s2 -L x1 -C(O)Q-L'-QC(O)-L x1 -, and each variable is independently as described in this disclosure. s2 is —CHC(O)Q-L′-QC(O)CH—, where each —CH— is independently optionally substituted. s2 is -CH2C(O)Q-L'-QC(O)CH2-.

[0214] In some embodiments, L s2 is L and contains at least one -C(O)-. In some embodiments, L s2 is L and contains at least two -C(O)-. In some embodiments, L s2 is L and comprises at least one -C(O)Q-, where Q is selected from the group consisting of a covalent bond, -N(R')-, -O-, and -S-. In some embodiments, L s2 is L and contains at least one -C(O)Q-, where Q is selected from -N(R')- and -O-. s2 is L and comprises at least two -C(O)Q-, where Q is selected from the group consisting of -N(R')-, -O-, and -S-. s2 is L and comprises at least two -C(O)Q-, where Q is selected from -N(R')- and -O-. s2 is L and contains at least one —C(O)N(R′)—. In some embodiments, L s2 is L and contains at least two -C(O)N(R')-. In some embodiments, L s2 is L and contains at least one -C(O)O-. In some embodiments, L s2is L and contains at least two -C(O)O-.

[0215] In some embodiments, L s2 includes -Q-L'-Q-, where Q is independently selected from the group consisting of -N(R')-, -O-, and -S, and L' is described in this disclosure.

[0216] In some embodiments, L s2 includes -Q-L'-Q-, where Q is independently selected from -N(R')- and -O-, and L' is described in this disclosure. s2 includes -C(O)Q-L'-QC(O)-, where Q is independently selected from the group consisting of -N(R')-, -O-, and -S, and L' is described in this disclosure. s2 includes -C(O)Q-L'-QC(O)-, where Q is independently selected from -N(R')- and -O, and L' is described in this disclosure. s2 comprises -C(R')C(O)Q-L'-QC(O)C(R')-, where Q is independently selected from the group consisting of -N(R')-, -O-, and -S, and L' is described in this disclosure. s2 includes -C(R')C(O)Q-L'-QC(O)C(R')-, where Q is independently selected from -N(R')- and -O, and L' is described in this disclosure.

[0217] In some embodiments, L s2 includes -N(R')-L'-N(R')-, where L' is described in this disclosure. s2 includes -C(O)N(R')-L'-N(R')C(O)-, where L' is described in this disclosure. s2 includes —C(R′)C(O)N(R′)-L′-N(R′)C(O)C(R′)—, where L′ is described in this disclosure.

[0218] In some embodiments, L s2includes -O(R')-L'-O(R')-, where L' is described in this disclosure. s2 includes -C(O)O-L'-OC(O)-, where L' is described in this disclosure. s2 includes —C(R′)2C(O)O-L′-OC(O)C(R′)2-, where L′ is described in this disclosure.

[0219] In some embodiments, R' is optionally substituted C 1~30 In some embodiments, R' is an optionally substituted C 1~15 In some embodiments, R' is an optionally substituted C 1~10 In some embodiments, R' is an optionally substituted C 1~5 It is aliphatic. In some embodiments, R' is hydrogen.

[0220] In some embodiments, L' is an optionally substituted divalent C1-C 19 In some embodiments, L' is an optionally substituted divalent C1-C 15 In some embodiments, L' is an optionally substituted divalent C1-C 10In some embodiments, L' is an optionally substituted divalent C1-C9 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C8 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C7 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C6 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C5 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C3 aliphatic. In some embodiments, L' is an optionally substituted divalent C1-C2 aliphatic. In some embodiments, L' is an optionally substituted divalent C1 aliphatic. In some embodiments, L' is -CH2-. In some embodiments, L' is -(CH2)2-. In some embodiments, L' is -(CH2)3-. In some embodiments, L' is -(CH2)4-. In some embodiments, L' is -(CH2)5-. In some embodiments, L' is -(CH2)6-. In some embodiments, L' is -(CH2)7-. In some embodiments, L' is -(CH2)8-.

[0221] In some embodiments, L' is an optionally substituted divalent C 6~20 In some embodiments, L' is an optionally substituted divalent C 6~14 In some embodiments, L' is an optionally substituted divalent C 6~10In some embodiments, L' is an optionally substituted divalent C6 aryl ring. In some embodiments, L' is a divalent C6 aryl substituted with at least one halogen. In some embodiments, L' is a divalent C6 aryl substituted with at least two halogens. In some embodiments, L' is a divalent C6 aryl substituted with at least three halogens. In some embodiments, L' is a divalent C6 aryl substituted with four halogens. In some embodiments, L' is a divalent C6 aryl substituted with at least one fluorine. In some embodiments, L' is a divalent C6 aryl substituted with at least two fluorines. In some embodiments, L' is a divalent C6 aryl substituted with at least three fluorines. In some embodiments, L' is a divalent C6 aryl substituted with four fluorines. In some embodiments, L' is a divalent C6 aryl substituted with at least one chlorine. In some embodiments, L' is a divalent C6 aryl substituted with at least two chlorines. In some embodiments, L' is a divalent C6 aryl substituted with at least three chlorines. In some embodiments, L' is a divalent C6 aryl substituted with four chlorines. In some embodiments, L' is at least one -O(CH2) 0~4 In some embodiments, L' is a divalent C6 aryl substituted with at least two -O(CH2) 0~4 In some embodiments, L' is a divalent C6 aryl substituted with at least three -O(CH2) 0~4 In some embodiments, L' is a divalent C6 aryl substituted with four -O(CH2) 0~4 It is a divalent C6 aryl substituted with CH3.

[0222] In some embodiments, L' is a divalent 5- to 20-membered heteroaryl ring having 1 to 10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, L' is a divalent 5- to 6-membered heteroaryl ring having 1 to 4 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon. In some embodiments, L' is a divalent 5- to 6-membered heteroaryl ring having 1 to 4 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, L' is a divalent 6-membered heteroaryl ring having 1 to 2 heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, L' is a divalent 6-membered heteroaryl ring having 2 nitrogen atoms.

[0223] In some embodiments, L' is an optionally substituted divalent C 3~20 In some embodiments, L' is an optionally substituted divalent C 3~15 In some embodiments, L' is an optionally substituted divalent C 3~10 In some embodiments, L' is an optionally substituted divalent C 3~9 In some embodiments, L' is an optionally substituted divalent C 3~8 In some embodiments, L' is an optionally substituted divalent C 3~7 In some embodiments, L' is an optionally substituted divalent C 3~6 In some embodiments, L' is an optionally substituted divalent C 3~5 In some embodiments, L' is an optionally substituted divalent C 3~4In some embodiments, L' is an optionally substituted divalent C3 cycloaliphatic ring. In some embodiments, L' is an optionally substituted divalent C4 cycloaliphatic ring. In some embodiments, L' is an optionally substituted divalent C5 cycloaliphatic ring. In some embodiments, L' is an optionally substituted divalent C5 cycloalkyl ring. In some embodiments, L' is an optionally substituted divalent C5 cycloalkenyl ring. In some embodiments, L' is an optionally substituted divalent C6 cycloaliphatic ring. In some embodiments, L' is an optionally substituted divalent C6 cycloalkyl ring.

[0224] In some embodiments, L s2 comprises -N(R')-L'-N(R')-, where L' is a covalent bond. s2 includes -N(R)-N(R)-, where Each R is independently -H or C 1~30 C having 1 to 10 heteroatoms independently selected from aliphatic, oxygen, nitrogen, sulfur, phosphorus, and silicon 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 C having 1 to 10 heteroatoms independently selected from arylaliphatic, oxygen, nitrogen, sulfur, phosphorus, and silicon 6~30 an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon, and 3-30 membered heterocyclyl having 1-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon; or Two or more R groups on two or more atoms optionally independently combine with their intervening atoms to form an optionally substituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having, in addition to the intervening atoms, 0-10 heteroatoms independently selected from oxygen, nitrogen, sulfur, phosphorus, and silicon.

[0225] In some embodiments, L s2 includes -N(R)-N(R)-, where Each R independently represents an optionally substituted C 1~30 aliphatic, or Two or more R groups on two or more atoms, optionally independently, together with their intervening atoms, form an optionally substituted 3- to 30-membered monocyclic ring.

[0226] In some embodiments, L s2 teeth, [ka] The staple is selected from the group consisting of:

[0227] In some embodiments, L s1 is an optionally substituted divalent C 1~6 In some embodiments, L s1 is a divalent C 1~6 In some embodiments, L s1 is a divalent C 1~4 In some embodiments, L s1 is saturated. In some embodiments, L s1 is linear. In some embodiments, L s1 is branched. In some embodiments, L s1 is optionally substituted —CH—. In some embodiments, L s1 is -CH-. In some embodiments, L s1 is optionally substituted -CH-CH-. In some embodiments, L s1 is -CH-CH-. In some embodiments, L s1 is optionally substituted —C(CH)—. In some embodiments, L s1 is -C(CH3)2-.

[0228] In some embodiments, L s2 is an optionally substituted divalent C1~6 (For example, C 3~6 , C3, C4, C5, C6, etc.) aliphatic, wherein one or more methylene units are optionally independently replaced with -Cy- or -C(R')2-. s2 is an optionally substituted divalent C 1~6 In some embodiments, L s2 is an optionally substituted divalent C 3~6 In some embodiments, L s2 is a divalent C 1~6 In some embodiments, L s2 is a divalent C 1~4 In some embodiments, L s2 is an optionally substituted divalent C2 aliphatic. s2 is an optionally substituted divalent C3 aliphatic. In some embodiments, L s2 is an optionally substituted divalent C4 aliphatic. In some embodiments, L s2 is an optionally substituted divalent C5 aliphatic. In some embodiments, L s2 is an optionally substituted divalent C6 aliphatic. In some embodiments, L s2 is substituted. In some embodiments, L s2 is unsubstituted. In some embodiments, L s2 is saturated. In some embodiments, L s2 is linear. In some embodiments, L s2 is branched. In some embodiments, L s2 is an optionally substituted divalent C 3~6 (For example, C 3~5 , C3, C4, C5, C6, etc.) aliphatic, wherein one or two methylene units are independently replaced with -Cy-. s2 is -CH2-Cy-CH2-. In some embodiments, L s2 is -CH-CH-Cy-CH-CH-. In some embodiments, L s2is -CH2-Cy-Cy-CH2-. Various useful embodiments of -Cy- are as described herein. For example, in some embodiments, -Cy- is an optionally substituted monocyclic 5-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy- is an optionally substituted monocyclic 6-membered aromatic ring having 0 to 4 heteroatoms. In some embodiments, -Cy- is optionally substituted phenylene. In some embodiments, -Cy- is optionally substituted 1,2-phenylene. In some embodiments, -Cy- is 1,2-phenylene. In some embodiments, -Cy- is optionally substituted 1,3-phenylene. In some embodiments, -Cy- is 1,3-phenylene. In some embodiments, -Cy- is optionally substituted 1,5-phenylene. In some embodiments, -Cy- is 1,5-phenylene. In some embodiments, -Cy- is 3-methyl-1,5-phenylene. In some embodiments, -Cy- is 3-methoxy-1,5-phenylene. In some embodiments, -Cy- is an optionally substituted bivalent pyridyl ring. In some embodiments, -Cy- is an optionally substituted [ka] In some embodiments, -Cy- is [ka] In some embodiments, -Cy- is optionally substituted [ka] In some embodiments, -Cy- is [ka] In some embodiments, -Cy- is an optionally substituted bicyclic 9-membered aromatic ring having 0-4 heteroatoms. In some embodiments, -Cy- is an optionally substituted bicyclic 10-membered aromatic ring having 0-4 heteroatoms. In some embodiments, -Cy- is an optionally substituted divalent naphthyl ring. In some embodiments, -Cy- is a divalent naphthyl ring. In some embodiments, -Cy- is an optionally substituted [ka] In some embodiments, -Cy- is [ka] In some embodiments, -Cy- is optionally substituted [ka] In some embodiments, -Cy- is [ka] In some embodiments, -Cy- is an optionally substituted 3- to 10-membered (e.g., 5-10, 5-6, 3, 4, 5, 6, 7, 8, 9, 10-membered, etc.) divalent alicyclic ring. In some embodiments, -Cy- is saturated. In some embodiments, -Cy- is an optionally substituted 6-membered cycloalkyl ring. In some embodiments, -Cy- is an optionally substituted [ka] In some embodiments, -Cy- is [ka] In some embodiments, L s2 is an optionally substituted divalent C 3~6 (For example, C 3~5, C3, C4, C5, C6, etc.) aliphatic, wherein one or two methylene units are independently replaced with -C(R')2-. s2 is -CH2-C(R')2-CH2-. In some embodiments, two R's taken together with the carbon atoms form an optionally substituted ring as described herein, for example, an optionally substituted 3-10 membered (e.g., 5-10, 5-6, 3, 4, 5, 6, 7, 8, 9, 10 membered, etc.) ring having 0-4 (e.g., 1-4, 0, 1, 2, 3, 4, etc.) heteroatoms. In some embodiments, the ring is saturated. In some embodiments, the ring has one or more heteroatoms. In some embodiments, -C(R')2- is [ka] is.

[0229] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, Ls2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is optionally substituted —(CH 2 ) 4 —. In some embodiments, L s2 is optionally substituted —(CH)—. In some embodiments, L s2 is optionally substituted -CH-CH=CH-CH-. In some embodiments, L s2 is optionally substituted (E)-CH-CH=CH-CH-. In some embodiments, L s2 is optionally substituted —CH—C(O)—CH—. In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is replaced as necessary [ka] In some embodiments, L s2 is substituted. In some embodiments, L s2 is unsubstituted. In some embodiments, [ka] , -(CH2)4-, (E)-CH2-CH=CH-CH2-, -(CH2)3-, and / or -CH2-C(O)-CH2- under equivalent conditions, [ka] provides higher binding and / or efficacy than

[0230] In some embodiments, L s3 is an optionally substituted divalent C 1~6 In some embodiments, L s3 is a divalent C 1~6 In some embodiments, L s3 is a divalent C 1~4 In some embodiments, L s3 is saturated. In some embodiments, L s3 is linear. In some embodiments, L s3 is branched. In some embodiments, L s3 is optionally substituted —CH—. In some embodiments, L s3 is -CH-. In some embodiments, L s3 is optionally substituted -CH-CH-. In some embodiments, L s3is -CH-CH-. In some embodiments, L s3 is optionally substituted —C(CH)—. In some embodiments, L s3 is -C(CH3)2-.

[0231] In some embodiments, the amino acid residues to form the staple are: [ka] In some embodiments, both amino acid residues forming the staple are independently selected from these amino acid residues. s1 and L s3 is independently -CH2-, -CH2-CH2-, or -C(CH3)2-. In some embodiments, the staple is formed by reacting a thiol group with a thiol-reactive linker compound. In some embodiments, such a linker compound is LG-L s2 -LG, or a salt thereof, wherein each LG is independently a leaving group, e.g., -Br, -I, etc. In some embodiments, each LG is independently -Br or -I. In some embodiments, each LG is -Br. In some embodiments, each LG is -I. In some embodiments, L s2 is LG-L s2 -LG, where each LG is independently -Br or -I; [ka] of such structure is a compound selected from

[0232] Various techniques are available for constructing thioether staples. For example, in some embodiments, the peptide and an excess of equivalents (e.g., about 2-10, 5-10, 2, 3, 4, 5, 6, 7, 8, 9, 10 equivalents, etc.; in some embodiments, 5 equivalents) of linker compound are added to a 1:1 DMF:100 mM Na2CO3 pH 8.0 solution and stirred at a suitable temperature, e.g., room temperature, for a suitable period of time, in some embodiments, 1-2 hours. In some embodiments, for example, for relatively weak electrophiles, an excess equivalent (e.g., about 10-30, 10-20, 10, 20, etc.; in some embodiments, 20 equivalents) of a metal salt, e.g., Zn(acac)2, and an excess equivalent (e.g., about 5-20, 10-15, 10, 15, 20, etc.; in some embodiments, 10-15 equivalents) of a linker compound are added to the peptide in DMA, and the mixture is stirred for a suitable period of time, e.g., overnight, at a suitable temperature, e.g., 37°C. In some embodiments, the equivalents of Zn(acac)2 and linker compound are doubled, and / or the temperature is increased to 50°C. In some embodiments, certain linker compounds react better than others. For example, in some embodiments, [ka] provide poor reaction yields or reaction failures. Other techniques can be used to obtain the corresponding linker moiety (L), for example, by using other leaving groups or by other reaction mechanisms / pathways. s2 It will be understood by those skilled in the art that a

[0233] In some embodiments, -L s1 -SL s2 -SL s3 A staple having the structure - is an (i, i+4) staple. In some embodiments, such a staple is closer to the C-terminus. In some embodiments, such a staple is closer to the N-terminus. For example, in some embodiments, such a staple is 10 and X 14 It is between.

[0234] In some embodiments, certain staples provide better properties and / or activity. For example, in some embodiments, based on target binding affinity, certain staples / scaffolds are ranked in the following order: [ka]

[0235] Those skilled in the art will recognize that the provided techniques can be utilized to prepare collections of peptides using non-cysteine ​​residues and suitable chemical reactions therefor. For example, in some embodiments, cysteine ​​stapling is replaced with lysine stapling, where the cysteine ​​residue for cysteine ​​stapling is replaced with a lysine residue for lysine stapling (e.g., using an agent capable of cross-linking two lysine residues, e.g., through reaction with side chain amino groups). In some embodiments, for lysine stapling, R in various formulas can be replaced with Eis or includes an activated carboxylic acid group (e.g., an NHS ester group), an imidoester group, or the like. Suitable reagents are widely known in the art, including many commercially available ones. In some embodiments, cysteine ​​stapling is replaced with methionine stapling. In some embodiments, a cysteine ​​residue for cysteine ​​stapling is replaced with a methionine residue for methionine stapling. In some embodiments, cysteine ​​stapling is replaced with a tryptophan residue for tryptophan stapling. Those skilled in the art will recognize that various techniques (e.g., reagents, reactions, etc.) have been described in the art and can be utilized in accordance with the present disclosure, e.g., for methionine stapling, tryptophan stapling, etc. In some embodiments, such stapling can be performed using reagents having various formulas described herein, where R E is or includes a group suitable for methionine and / or tryptophan stapling. In some embodiments, stapling can be performed using one residue at the first position and a different residue at the second position. Reagents useful for such stapling include those suitable for stapling at the first position (e.g., the first R E a first reactive group for stapling at the second position (e.g., via a second R E The compound may include a second reactive group for reaction with the hydroxyl group (via

[0236] In some embodiments, for various types of stapling (e.g., cysteine ​​or non-cysteine ​​stapling), stapling is between residues (e.g., cysteine ​​residues for cysteine ​​stapling) separated by two residues (i+3 stapling). In some embodiments, stapling is between residues separated by three residues (i+4 stapling). In some embodiments, stapling is between residues separated by six residues (i+7 stapling).

[0237] As will be appreciated by those skilled in the art, in some embodiments, more than two residues may be stapled simultaneously. For example, in some embodiments, three or more cysteines may be stapled with three or more reactive groups (e.g., R E The polymer is stapled using a cross-linking reagent containing a .alpha.-group.

[0238] In some embodiments, as described herein, the present disclosure provides useful techniques related to non-cysteine ​​stapling. In particular, the present disclosure recognizes that peptides suitable for cysteine ​​stapling and / or containing one or more non-cysteine ​​staples may have their cysteine ​​residues and cysteine ​​staples replaced with other amino acids and staples described herein (e.g., hydrocarbon and other non-hydrocarbon amino acids and staples). In some embodiments, the resulting non-cysteine ​​stapled peptide maintains the same or similar interaction with a target of interest compared to a reference cysteine ​​stapled peptide.

[0239] Certain useful agents (pre-stapling peptides and post-stapling staple peptides) and compositions thereof are presented as examples in Table E2 or Table E3, including various amino acid residues and N- and C-terminal capping groups at various positions, as well as various stapling patterns, e.g., X 1 -X4 -X 11 , X 1 -X 3 , X 3 -X 7 , X 3 -X 10 , X 4 -X 11 , X 7 -X 10 , X 7 -X 14 , X 10 -X 14 As demonstrated herein, the techniques provided can deliver improved useful properties and / or activities.

[0240] In some embodiments, the provided agent, peptide, or staple peptide is a compound described herein. In some embodiments, the provided agent has a structure selected from Table E2 or Table E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer of a structure selected from Table E2 or Table E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer with respect to a chiral center attached to two staples (e.g., with respect to B4, B5, etc.) of a structure selected from Table E2 or Table E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer with respect to an olefinic double bond in a staple of a structure selected from Table E2 or Table E3, or a salt thereof. In some embodiments, the provided agent is a stereoisomer with respect to an olefinic double bond and / or a chiral center attached to two staples (e.g., with respect to B4, B5, etc.) of a staple of a structure selected from Table E2 or Table E3, or a salt thereof. In some embodiments, the provided composition is a composition described in Table E2 or Table E3. In some embodiments, the compound is [ka] (SP-1) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-2) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-3) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-4) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-5) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-6) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-7) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-8) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-9) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-10) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-11) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-12) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-13) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-14) or a salt thereof. In some embodiments, the compound has the structure: [ka] (SP-15) or a salt thereof. In some embodiments, the double bond of the (i, i+2) staple is E. In some embodiments, the double bond of the (i, i+2) staple is Z. In some embodiments, the double bond of the (i, i+3) staple is E. In some embodiments, the double bond of the (i, i+3) staple is Z. In some embodiments, the double bond of the (i, i+7) staple is E. In some embodiments, the double bond of the (i, i+7) staple is Z. In some embodiments, both double bonds are E. In some embodiments, both double bonds are Z. In some embodiments, the (i, i+3) staple is E and the other is Z. In some embodiments, the (i, i+3) staple is Z and the other is E. In some embodiments, the (i, i+4) staple is E and the other is Z. In some embodiments, the (i, i+4) staple is Z and the other is E. In some embodiments, the double bond of the (i, i+7) staple is Z and the double bond of the second staple (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is E. In some embodiments, the double bond of the (i, i+7) staple is Z and the double bond of the second staple (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is Z. In some embodiments, the double bond of the (i, i+7) staple is E and the double bond of the second staple (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is E. In some embodiments, the double bond of the (i, i+7) staple is E and the double bond of the second staple (e.g., (i, i+2), (i, i+3), (i, i+4), etc.) is Z. In some embodiments, two staples are attached to a chiral center (e.g., a carbon atom in B5) and the chiral center is R. In some embodiments, two staples are attached to a chiral center (e.g., a carbon atom in B5) and the chiral center is S.

[0241] In some embodiments, the compound has a structure selected from below, or a salt thereof: [ka]

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[0242] Agents, e.g., peptides, including stapled peptides, can contain a variety of numbers of amino acid residues. In some embodiments, the length of a peptide agent is about 5-20, 5-19, 5-18, 5-17, 5-16, 5-15, 10-20, 10-19, 10-18, 10-17, 10-16, 10-15, 11-20, 11-19, 11-18, 11-17, 11-16, 11-15, 12-20, 12-19, 12-18, 12-17, 12-16, The length may be 12-15, 13-20, 13-19, 13-18, 13-17, 13-16, 13-15, 14-20, 14-19, 14-18, 14-17, 14-16, 14-15, or about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues. In some embodiments, the length is about 10 amino acid residues. In some embodiments, the length is about 11 amino acid residues. In some embodiments, the length is about 12 amino acid residues. In some embodiments, the length is about 13 amino acid residues. In some embodiments, the length is about 14 amino acid residues. In some embodiments, the length is about 15 amino acid residues. In some embodiments, the length is about 16 amino acid residues. In some embodiments, the length is about 17 amino acid residues. In some embodiments, the length is about 18 amino acid residues. In some embodiments, the length is about 19 amino acid residues. In some embodiments, the length is about 20 amino acid residues.

[0243] In some embodiments, as described herein, one or more staples independently comprise an olefinic double bond (e.g., formed by olefin metathesis). In some embodiments, one or more staples independently comprise an amide group (e.g., formed by amidation). In some embodiments, at least one staple does not contain an olefinic double bond. In some embodiments, there is at least one staple whose formation does not involve a reaction of an olefin, such as olefin metathesis and / or modification of an olefinic double bond (e.g., hydrogenation, epoxidation, etc.).

[0244] In some embodiments, the residue of the staple (e.g., B5) is selected from those residues at which the position is P (e.g., X 4 ), the first acidic amino acid residue is at the P-2 position (e.g., X 2 ), and a second acidic amino acid residue is located at P+1 (e.g., X 5 ), and a third acidic amino acid residue is located at P+2 (e.g., X 6 ), and a hydrophobic amino acid residue is located at P+4 (e.g., X 8 ), and the first aromatic amino acid residue is located at P+5 (e.g., X 9 ), and a second aromatic amino acid residue is located at P+8 (e.g., X 12 ), and / or a third aromatic amino acid residue is located at P+9 (e.g., X 13 In some embodiments, the staple is a (i, i+7) staple, and the other residue of the staple is located at P+7 (e.g., X 11 In some embodiments, the first acidic amino acid residue is located at P-2 (e.g., X 2 In some embodiments, the second acidic amino acid residue is located at P+1 (e.g., X 5 In some embodiments, the third acidic amino acid residue is located at P+2 (e.g., X 6 In some embodiments, the hydrophobic amino acid residue is located at P+4 (e.g., X 8 In some embodiments, the first aromatic amino acid residue is located at P+5 (e.g., X9 In some embodiments, the second aromatic amino acid residue is located at P+8 (e.g., X 12 In some embodiments, the third aromatic amino acid residue is located at P+9 (e.g., X 13 In some embodiments, the first acidic amino acid residue is located at P-2 (e.g., X 2 ), and the second acidic amino acid residue is located at P+1 (e.g., X 5 ), and the first aromatic amino acid residue is located at P+5 (e.g., X 9 ), and the second aromatic amino acid residue is located at P+8 (e.g., X 12 ), the third aromatic amino acid residue is located at P+9 (e.g., X 13 In some embodiments, the first acidic amino acid residue is at the P-2 position (e.g., X 2 ), and the second acidic amino acid residue is located at P+1 (e.g., X 5 ), and the third acidic amino acid residue is located at P+2 (e.g., X 6 ), and a hydrophobic amino acid residue is located at P+4 (e.g., X 8 ), and the first aromatic amino acid residue is located at P+5 (e.g., X 9 ), and the second aromatic amino acid residue is located at P+8 (e.g., X 12 ), the third aromatic amino acid residue is located at P+9 (e.g., X 13). In some embodiments, a stapled peptide agent comprises acidic amino acid residues at the P-2 and P+1 positions, and aromatic amino acid residues at the P+5, P+8, and P+9 positions. In some embodiments, a stapled peptide agent comprises acidic amino acid residues at the P-2, P+1, and P+2 positions, and aromatic amino acid residues at the P+5, P+8, and P+9 positions. In some embodiments, a stapled peptide agent comprises acidic amino acid residues at the P-2 and P+1 positions, a hydrophobic amino acid residue at the P+4 position, and aromatic amino acid residues at the P+5, P+8, and P+9 positions. In some embodiments, a stapled peptide agent comprises acidic amino acid residues at the P-2, P+1, and P+2 positions, a hydrophobic amino acid residue at the P+4 position, and aromatic amino acid residues at the P+5, P+8, and P+9 positions. In some embodiments, a stapled peptide agent comprises acidic amino acid residues at the P-2, P+1, and P+2 positions, a hydrophobic amino acid residue at the P+4 position, and aromatic amino acid residues at the P+5, P+8, and P+9 positions. In some embodiments, P is 3. In some embodiments, P is 4. In some embodiments, P is 5. In some embodiments, P is 6. In some embodiments, P is 7. In some embodiments, the amino acid residue at position P includes two groups for stapling, e.g., B4, B5, B6, etc. In some embodiments, the group is B4. In some embodiments, the group is B5. In some embodiments, the group is B6. In some embodiments, the agent includes a staple and a first additional staple, e.g., an (i, i+3) or (i, i+4) staple. In some embodiments, the staple and the first additional staple are attached to the same residue (e.g., B5, B6, etc.). In some embodiments, the other residue of the first additional residue is at position P-2 (e.g., if a moiety for stapling, such as a terminal olefin, is at the P terminal group, this is X 1 In some embodiments, the agent is at a second additional staple, e.g., at the (i, i+4) staple (e.g., at the P+6 position (e.g., X 10 ) and P+10 positions (e.g., X 14 ) stapling residues), (i, i+3) staples (e.g., P+3 positions (e.g., X 7 ) and P+6 position (e.g., X 10) stapling residues), (i, i+7) staples (e.g., P+3 positions (e.g., X 7 ) and P+10 positions (e.g., X 14 In some embodiments, the agent comprises a second additional staple, which is located at the P+6 position of the (i, i+4) staple (e.g., X 10 ) and P+10 positions (e.g., X 14 In some embodiments, the agent is a stapling residue at the P-1 position (e.g., X) of a third additional staple, e.g., the (i, i+4) staple. 3 ) and P+3 positions (e.g., X 7 ) stapling residues. In some embodiments, there are three staples in the stapled peptide agent. In some embodiments, there are four staples in the stapled peptide agent. As demonstrated herein, stapled agents comprising staples and residues so positioned can provide a variety of desirable properties and activities. In some embodiments, the positioning of one or more staples may be shifted relative to the various acidic, hydrophobic, and / or aromatic amino acid residues described herein; for example, in some embodiments, the stapled peptide agent comprises staple residues at positions P and P+7 (and optionally P-3 or P-4), with acidic amino acid residues at positions P-1 and P+2, aromatic amino acid residues at positions P+6, P+9, and P+10, and optionally an acidic amino acid residue at position P+3 and / or a hydrophobic amino acid residue at position P+5. It has been observed that various stapled peptide agents with shifted staples are capable of binding to beta-catenin as assessed by fluorescence polarization.

[0245] Certain useful staples are described below in the "Medications" section. beta-catenin

[0246] In particular, the present disclosure provides techniques for modulating one or more beta-catenin functions. In some embodiments, the present disclosure provides useful techniques for inhibiting one or more beta-catenin functions associated with cancer or hyperplasia. In some embodiments, the provided techniques are useful for preventing and treating conditions, disorders, or diseases that would benefit from the prevention and / or treatment of beta-catenin inhibition. In some embodiments, the condition, disorder, or disease is cancer.

[0247] Beta-catenin has been reported to have various functions. For example, it can regulate and coordinate the transcription of various genes. It has been reported that high beta-catenin activity and / or expression levels can contribute to the development of various conditions, disorders, or diseases, including cancer. Mutations and overexpression of beta-catenin have been reported to be associated with conditions, disorders, or diseases, including numerous cancers, including colorectal cancer, lung cancer, and breast cancer. Dysregulation of the Wnt / beta-catenin signaling pathway has been reported to be associated with several conditions, disorders, or diseases, including neurodegenerative diseases, psychiatric disorders, cancer, asthma, and even wound healing. Numerous published studies, both clinical and preclinical, have shown that overactivated Wnt / beta-catenin activity drives tumorigenesis and is required for tumor maintenance in various cancers. Many Wnt inhibitors primarily modulate this pathway at the extracellular ligand / receptor level, for example, by preventing Wnt ligand secretion or by blocking Wnt ligand interaction with its receptor at the plasma membrane. Many Wnt pathway-activating mutations have been reported to be found in APC and / or CTNNB1, downstream of membrane-proximal events. In particular, the present disclosure encompasses the recognition that many drugs at the extracellular ligand / receptor level are inadequate to treat many related patients, for example, patients with downstream mutations / abnormalities. In some embodiments, Wnt pathway-activating mutations are concentrated in the beta-catenin / TCF node. In some embodiments, the present disclosure targets beta-catenin / TCF interactions, for example, as a therapeutic approach. Agents that can modulate beta-catenin function are useful for various purposes, including preventing and / or treating various conditions, disorders, or diseases associated with beta-catenin. binding site

[0248] Beta-catenin can interact with various drugs through various binding sites, each of which independently comprises a set of amino acid residues that interact with binding agents.For example, a specific binding site is used for the interaction of beta-catenin with axin, APC, C-cadherin, E-cadherin, TCF3 and Bcl9.For the interaction with TCF3, it has been reported that two or more binding sites can be simultaneously used to interact with different parts of TCF3.For example, see Graham et al.Cell, Vol.103, 885-896, 2000.

[0249] In some embodiments, provided agents bind to beta-catenin at a unique binding site, ie, provided agents interact with beta-catenin at a set of amino acid residues that differs from previously reported binding sites, e.g., for Axin, APC, C-cadherin, E-cadherin, TCF3, or Bcl9.

[0250] For example, in some embodiments, provided agents may be selected from the amino acid residues in SEQ ID NO: 1, such as, in some embodiments, the following amino acid residues of SEQ ID NO: 1: A305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D and one or more or all of a set of amino acid residues that are or correspond to: N413, N415, V416, T418, and C419 (e.g., about 1-23, 1-20, 1-15, 1-10, 1-5, 5-23, 5-20, 5-15, 5-10, 6-23, 6-20, 6-15, 6-10, 7-23, 7-2 0 pieces, 7~15 pieces, 7~10 pieces, 8~23 pieces, 8~20 pieces, 8~15 pieces, 8~10 pieces, 9~23 pieces, 9~20 pieces, 9~15 pieces, 9~10 pieces, 10~23 pieces, 10~20 pieces, 10~ 15 pieces, 11~23 pieces, 11~20 pieces, 11~15 pieces, 12~23 pieces, 12~20 pieces, 12~15 pieces, 13~23 pieces, 13~20 pieces, 13~15 pieces, 13~23 pieces, 14~20 pieces, 15-23, 15-20, 16-23, 16-20, 17-23, 17-20, 18-23, or 18-20, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, or 23, etc. In some embodiments, the set of amino acid residues is or corresponds to amino acid residues A305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419 of SEQ ID NO:1. In some embodiments, the set of amino acid residues is or corresponds to amino acid residues A305, Y306, G307, N308, Q309, K312, K345, V346, V349, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419 of SEQ ID NO:1.In some embodiments, the set of amino acid residues are or correspond to amino acid residues G307, K312, K345, W383, N387, D413, and N415 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues are or correspond to amino acid residues G307, K312, K345, Q379, L382, W383, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues are or correspond to amino acid residues Y306, G307, K312, K345, Q379, L382, W383, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues are or correspond to amino acid residues G307, K312, K345, Q379, L382, W383, R386, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues are or correspond to amino acid residues Y306, G307, K312, K345, Q379, L382, W383, R386, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues are or correspond to amino acid residues Y306, G307, K312, K345, V349, Q379, L382, W383, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues is or corresponds to amino acid residues Y306, G307, K312, K345, V349, Q379, L382, W383, R386, N387, N415, and V416 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues is or corresponds to amino acid residues G307, K312, K345, W383, and N387 of SEQ ID NO: 1. In some embodiments, the set of amino acid residues is or corresponds to amino acid residues Y306, G307, K312, R386, and N387 of SEQ ID NO: 1. In some embodiments, provided agents interact with Y306 or the amino acid residue corresponding thereto. In some embodiments, provided agents interact with G307 or the amino acid residue corresponding thereto.In some embodiments, provided agents interact with K312 or a corresponding amino acid residue. In some embodiments, provided agents interact with K345 or a corresponding amino acid residue. In some embodiments, provided agents interact with V349 or a corresponding amino acid residue. In some embodiments, provided agents interact with Q379 or a corresponding amino acid residue. In some embodiments, provided agents interact with L382 or a corresponding amino acid residue. In some embodiments, provided agents interact with W383 or a corresponding amino acid residue. In some embodiments, provided agents interact with R386 or a corresponding amino acid residue. In some embodiments, provided agents interact with N387 or a corresponding amino acid residue. In some embodiments, provided agents interact with N415 or a corresponding amino acid residue. In some embodiments, provided agents interact with V416 or a corresponding amino acid residue.

[0251] In some embodiments, the agent interacts with a polypeptide whose sequence corresponds to aa 146 to aa 665 of human beta-catenin. In some embodiments, the agent interacts with a polypeptide whose sequence is or includes SEQ ID NO:2. SVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKMVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTSRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEGLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRT (SEQ ID NO: 2)

[0252] In some embodiments, all amino acid residues that interact with a provided agent are in SEQ ID NO: 2. In some embodiments, amino acid residues that interact with a provided agent (e.g., one or more amino acid residues in the agent) interact with the agent through hydrogen bonds, hydrophobic interactions, or salt bridges. As will be understood by those skilled in the art, when two amino acid residues interact with each other, they are typically within a certain distance range, as assessed, for example, using crystallography, NMR, etc.

[0253] In some embodiments, certain amino acid residues that have been reported to interact with one or more polypeptides are not significantly involved in the interaction between the provided agent and beta-catenin. In some embodiments, provided agents do not interact with the axin-binding site. In some embodiments, provided agents do not interact with the Bcl9-binding site. In some embodiments, provided agents do not interact with one or more or all of the amino acid residues at or corresponding to N426, C429, K435, R469, H470, S473, R474, K508, and N516 of SEQ ID NO: 1. In some embodiments, provided agents do not interact with N426 or the amino acid residue corresponding thereto. In some embodiments, provided agents do not interact with C429 or the amino acid residue corresponding thereto. In some embodiments, provided agents do not interact with K435 or the amino acid residue corresponding thereto. In some embodiments, provided agents do not interact with R469 or the amino acid residue corresponding thereto. In some embodiments, provided agents do not interact with H470 or the amino acid residue corresponding thereto. In some embodiments, provided agents do not interact with S473 or a corresponding amino acid residue. In some embodiments, provided agents do not interact with R474 or a corresponding amino acid residue. In some embodiments, provided agents do not interact with K508 or a corresponding amino acid residue. In some embodiments, provided agents do not interact with N516 or a corresponding amino acid residue.

[0254] In some embodiments, mutation of one or more amino acid residues in beta-catenin other than SEQ ID NO: 2 does not significantly (e.g., by more than 30%, more than 40%, more than 50%, more than 60%, more than 70%, more than 75%, more than 80%, more than 85%, more than 90%, or more) reduce the interaction of beta-catenin with the provided agent. In some embodiments, mutation of one or more or all of the amino acid residues at or corresponding to N426, C429, K435, R469, H470, S473, R474, K508, and N516 of SEQ ID NO: 1 does not significantly reduce the interaction of beta-catenin with the provided agent. In some embodiments, mutation of N426 or the amino acid residue corresponding thereto reduces the interaction of beta-catenin with the provided agent. In some embodiments, mutation of Q379 or a corresponding amino acid residue (e.g., Ala, Glu, Phe, Trp, etc.) does not significantly reduce the interaction of beta-catenin with the drug.

[0255] In some embodiments, the agent binds to the TCF site of beta-catenin. In some embodiments, the agent interacts with one or more, but not all, amino acid residues that interact with TCF. In some embodiments, the agent interacts with one or more, but not all, amino acid residues that interact with the extended region of XTcf3-CBD. In some embodiments, the agent does not interact with beta-catenin amino acid residues that interact with the beta-hairpin module of XTcf3-CBD. In some embodiments, the agent does not interact with beta-catenin amino acid residues that interact with the helix module of XTcf3-CBD. For certain amino acid residues that interact with various modules of XTcf3-CBD, see, for example, Graham et al. Cell, Vol. 103, 885-896, 2000.

[0256] In some embodiments, the agent competes with TCF for beta-catenin binding. In some embodiments, the agent inhibits an extended region of TCF (e.g., Ala14-Glu24 or Asp16-Glu24, Graham et al. Cell, Vol. 103, 885-896, 2000). In some embodiments, compared to the expanded region of the TCF, the agent does not compete with axin or competes to a lesser extent for beta-catenin binding. In some embodiments, compared to the expanded region of the TCF, the agent does not compete with Bcl9 or competes to a lesser extent for beta-catenin binding. In some embodiments, compared to the expanded region of the TCF, the agent does not compete with the beta-hairpin module of XTcf3-CBD for beta-catenin binding. In some embodiments, compared to the expanded region of the TCF, the agent does not compete with the helix module of XTcf3-CBD for beta-catenin binding or competes to a lesser extent. In some embodiments, the agent competes with E-cadherin for beta-catenin binding.

[0257] In some embodiments, the present disclosure provides a complex between a peptide (e.g., a polypeptide whose sequence is or includes SEQ ID NO: 1 or 2) and a provided agent. In some embodiments, in such a complex, the polypeptide and the provided agent interact with one or more or all of the amino acid residues described herein, and optionally do not interact with one or more or all of the amino acid residues described herein.

[0258] In some embodiments, the present disclosure provides a complex comprising a provided agent and a beta-catenin polypeptide or a portion thereof. In some embodiments, the portion comprises one or more or all of the interacting residues described herein. In some embodiments, the agent and the beta-catenin polypeptide or a portion thereof interact with each other at one or more or all of the interacting residues. Certain drugs

[0259] In some embodiments, the present disclosure provides a structure of Formula I R N -L P1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C , I or a salt thereof, wherein R N is a peptide, an amino protecting group, or R'-L RN - and L P1 , L P2 , L P3 , L P4 , L P5 , L P6 , and L P7 Each of these is independent. て, L, L P1 , L P2 , L P3 , L P4 , L P5 , L P6 , and L P7 but, Let's get together -L s a first R′ group and a second R′ group, which are bonded to the atom to which the first R′ group is bonded and to the atom to which the second R′ group is bonded, and Let's get together -L s -, which is bonded to the atom to which the third R' group is bonded and to the atom to which the fourth R' group is bonded. Including, Each L s But independently, -L s1 -L s2 -L s3 - and each L s1 , L s2 , and L s3 is independently L, L AA1 is an amino acid residue containing a side chain containing an acidic or polar group, L AA2 is an amino acid residue containing a side chain containing an acidic or polar group, L AA3 is an amino acid residue, L AA4 is an amino acid residue comprising a side chain containing an optionally substituted aromatic group; L AA5 is an amino acid residue comprising a side chain containing an optionally substituted aromatic group; L AA6 is an amino acid residue comprising a side chain containing an optionally substituted aromatic group; R C peptide, carboxyl protecting group, -L RC -R', -OL RC -R', or -N(R')-L RC -R', L RN and L RC each of which is independently L; Each L is independently a covalent bond or an optionally substituted divalent C1-C6 25 Aliphatic group or divalent C1-C with 1-10 heteroatoms 25a heteroaliphatic group in which one or more methylene units of the group are optionally independently replaced by -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-; each -Cy- is independently an optionally substituted divalent 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; each R' is independently -LR, -C(O)R, -COR, or -SOR; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30 an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or two R groups, optionally independently, taken together to form a covalent bond; or two or more R groups on the same atom, optionally independently, taken together with the atom to form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to the atom, 0-10 heteroatoms; or two or more R groups on two or more atoms optionally independently, taken together with their intervening atoms, form an optionally substituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms; A drug or a salt thereof is provided.

[0260] In some embodiments, the present disclosure provides a structure of Formula I R N -LP1 -L AA1 -L P2 -L AA2 -L P3 -L AA3 -L P4 -L AA4 -L P5 -L AA5 -L P6 -L AA6 -L P7 -R C , I or a salt thereof, R N is a peptide, an amino protecting group, or R'-L RN - and L P1 , L P2 , L P3 , L P4 , L P5 , L P6 , and L P7 Each of these is independent. て, L, L P1 , L P2 , L P3 , L P4 , L P5 , L P6 , and L P7 but, Let's get together -L s a first R′ group and a second R′ group, which are bonded to the atom to which the first R′ group is bonded and to the atom to which the second R′ group is bonded, and Let's get together -L s -, which is bonded to the atom to which the third R' group is bonded and to the atom to which the fourth R' group is bonded. Including, Each L s But independently, -L s1 -L s2 -L s3 - and each L s1 , L s2 , and L s3 is independently L, L AA1 But, L ARand the methylene unit is -C(R')(R AS )- and R AS But -L AS1 -R AA1 and R AA1 is -CO2R or -SO2R, L AA2 But, L AR and the methylene unit is -C(R')(R AS )- and R AS But -L AS2 -R AA2 and R AA2 is -CO2R or -SO2R, L AA3 But, L AR and the methylene unit is -C(R')(R AS )- and R AS But -L AS3 -R AA3 and R AA3 is R', L AA4 But, L AR and the methylene unit is -C(R')(R AS )- and R AS But -L AS4 -R AA4 and R AA4 is an optionally substituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms; L AA5 But, L AR and the methylene unit is -C(R')(R AS )- and R AS But -L AS5 -R AA5 and R AA5 is an optionally substituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms; L AA6 But, L AR and the methylene unit is -C(R')(R AS )- and R AS But -L AS6 -RAA6 and R AA6 is an optionally substituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms; R C peptide, carboxyl protecting group, -L RC -R', -OL RC -R', or -N(R')-L RC -R', L RN and L RC each of which is independently L; Each L AR are independently an optionally substituted divalent C1-C4 aliphatic group, and one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS )-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-, and L AS1 , L AS2 , L AS3 , L AS4 , L AS5 , and L AS6 Each of the following is independently L AS and Each R AS But independently, -L AS -R', Each L AS are independently optionally substituted divalent C1 to C 10 Aliphatic group or divalent C1-C with 1-5 heteroatoms 10a heteroaliphatic group in which one or more methylene units of the group are optionally independently replaced by -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-; Each L is independently a covalent bond or an optionally substituted divalent C1-C6 25 Aliphatic group or divalent C1-C with 1-10 heteroatoms 25 a heteroaliphatic group in which one or more methylene units of the group are optionally independently replaced by -C(R')2-, -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-; each -Cy- is independently an optionally substituted divalent 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms; each R' is independently -LR, -C(O)R, -COR, or -SOR; Each R is independently -H or C 1~30 Aliphatic, C with 1-10 heteroatoms 1~30 Heteroaliphatic, C 6~30 Aryl, C 6~30 Arylaliphatic, C with 1-10 heteroatoms 6~30an optionally substituted group selected from arylheteroaliphatic, 5-30 membered heteroaryl having 1-10 heteroatoms, and 3-30 membered heterocyclyl having 1-10 heteroatoms; or two R groups, optionally independently, taken together to form a covalent bond; or two or more R groups on the same atom, optionally independently, taken together with the atom to form an optionally substituted 3-30 membered monocyclic, bicyclic, or polycyclic ring having, in addition to the atom, 0-10 heteroatoms; or two or more R groups on two or more atoms optionally independently, taken together with their intervening atoms, form an optionally substituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0-10 heteroatoms in addition to the intervening atoms; A drug or a salt thereof is provided.

[0261] In some embodiments, the second R' group and the third R' group are bonded to the same atom. In some embodiments, none of the first, second, and fourth R' groups are bonded to the same atom. In some embodiments, none of the first, second, fourth, fifth, and sixth R' groups are bonded to the same atom. In some embodiments, none of the first, second, fourth, fifth, sixth, seventh, and eighth R' groups are bonded to the same atom. In some embodiments, each of the first, second, third, and fourth R' groups is independently bonded to a different atom. In some embodiments, each of the first, second, third, fourth, fifth, and sixth R' groups is independently bonded to a different atom. In some embodiments, each of the first, second, third, fourth, fifth, and sixth R' groups is independently bonded to a different atom. In some embodiments, each of the first, second, third, fourth, fifth, sixth, seventh, and eighth R' groups is independently bonded to a different atom.

[0262] In some embodiments, the compound of formula I is a stapled peptide described herein.

[0263] In some embodiments, each L s are independently a staple described herein. In some embodiments, Ls , for example, the L formed by the first and second R′ groups together. s has a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. Unless otherwise indicated, the distance between two connecting sites, e.g., L s , L, etc. is the shortest covalent bond connection from one moiety to the other. For example, the length of -CH-CH- is two atoms (-CC-), and the length of 1,3-phenylene is three atoms. In some embodiments, L s , for example, the L formed by the third and fourth R′ groups together. s has a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. s , for example, the L formed by the fifth and sixth R′ groups together. s has a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms. s , for example, the L formed by the seventh and eighth R′ groups together. s has a length of 5 to 20 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) atoms.

[0264] Those skilled in the art, upon reading this disclosure, will recognize that staples connecting two atoms with a longer distance, e.g., L sIt will be understood that a staple connecting two atoms with a shorter distance typically has a longer length than a staple connecting two atoms with a shorter distance, for example, an (i, i+7) staple typically has a longer length than an (i, i+3) or (i, i+4) staple. In some embodiments, the length is 5 atoms. In some embodiments, the length is 6 atoms. In some embodiments, the length is 7 atoms. In some embodiments, the length is 8 atoms. In some embodiments, the length is 9 atoms. In some embodiments, the length is 10 atoms. In some embodiments, the length is 11 atoms. In some embodiments, the length is 12 atoms. In some embodiments, the length is 13 atoms. In some embodiments, the length is 14 atoms. In some embodiments, the length is 15 atoms. In some embodiments, the length is 16 atoms. In some embodiments, the length is 17 atoms. In some embodiments, the length is 18 atoms. In some embodiments, the length is 19 atoms. In some embodiments, the length is 20 atoms. L P1

[0265] In some embodiments, L P1 is a covalent bond or an optionally substituted divalent C-C aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L P1is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P1 is or comprises an amino acid residue. P1 is or comprises a peptide.

[0266] In some embodiments, L P1 -[X] p -X 1 - is or contains p, X, and X 1 are each independently as described herein; and X 1 L AA1 In some embodiments, L P1 -X 1 - is or contains.

[0267] In some embodiments, L P1contains a -C(R')2- group, where one of the R' groups is the first of the four R' groups. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of an X 1 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. L AA1

[0268] In some embodiments, L AA1 is or comprises an amino acid residue. AA1 is or comprises an amino acid residue comprising a side chain containing an acidic or polar group. AA1 is an amino acid residue containing a side chain containing an acidic group.

[0269] In some embodiments, L AA1 L AR and the methylene unit is -C(R')(R AS )-, where each variable is independently as described herein. AA1 is an optionally substituted divalent C1-C6 (e.g., C1, C2, C3, C4, C5, or C6) aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA1 is an optionally substituted divalent C2-C4 aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R ASIn some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA1 is -N(R')-C(R')(R AS )—C(O)—, and each variable is independently as described herein. In some embodiments, L AA1 is -NH-C(R')(R AS )—C(O)—, and each variable is independently as described herein.

[0270] In some embodiments, L AS1 is, as described herein, L AS In some embodiments, R AA1 is —COR, where R is as described herein. In some embodiments, R is H. In some embodiments, L AA1 is an acidic amino acid residue, e.g., a residue such as Asp, Glu, etc. In some embodiments, L AA1 is, as described herein, X 2 is. L P2

[0271] In some embodiments, L P2 is a covalent bond or an optionally substituted divalent C-C aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L P2is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P2 is or comprises an amino acid residue. P2 is or comprises a peptide.

[0272] In some embodiments, L P2 is -[X]pX 4 [X]p'- or containing p, p', X, and X 4 and each of L is independently as described herein. P2 is -[X]pX 3 X 4 [X]p'- or containing it, and each of X, X 3 and X 4 are independently an amino acid residue, and each of p and p' is independently 0 to 10. P2 -X 3 X 4 - is or contains each X 3and X 4 are independently as described herein; and X 4 L AA2 is bonded to.

[0273] In some embodiments, L P2 contains a -C(R')2- group, where one of the R' groups is the second of the four R' groups and the other is the third. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of X 4 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 4 is the alpha carbon of

[0274] In some embodiments, L P2 In some embodiments, the methylene unit of X is replaced with -C(R')2-, and one of the R' groups is the second, fifth, or seventh R' group. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of X 3 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 3 In some embodiments, it is the alpha carbon of . In some embodiments, it is the second R' group. In some embodiments, it is the fifth R' group. In some embodiments, it is the seventh R' group.

[0275] In some embodiments, L P2 wherein the methylene unit of X is replaced with -C(R')2-, and one of the R' groups is the first or third R' group. In some embodiments, such -C(R')2- groups are those of amino acid residues. In some embodiments, such -C(R')2- groups are those of X 4 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 4In some embodiments, it is the alpha carbon of . In some embodiments, it is the first R' group. In some embodiments, it is the third R' group. L AA2

[0276] In some embodiments, L AA2 is or comprises an amino acid residue. AA2 is or comprises an amino acid residue comprising a side chain containing an acidic or polar group. AA2 is an amino acid residue containing a side chain containing an acidic group.

[0277] In some embodiments, L AA2 L AR and the methylene unit is -C(R')(R AS )-, where each variable is independently as described herein. AA2 is an optionally substituted divalent C1-C6 (e.g., C1, C2, C3, C4, C5, or C6) aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA2 is an optionally substituted divalent C2-C4 aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R ASIn some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA2 is -N(R')-C(R')(R AS )—C(O)—, and each variable is independently as described herein. In some embodiments, L AA2 is -NH-C(R')(R AS )—C(O)—, and each variable is independently as described herein.

[0278] In some embodiments, L AS2 is, as described herein, L AS In some embodiments, R AA2 is —COR, where R is as described herein. In some embodiments, R is H. In some embodiments, L AA2 is an acidic amino acid residue, e.g., a residue such as Asp, Glu, etc. In some embodiments, L AA2 is, as described herein, X 5 is. L P3

[0279] In some embodiments, L P3 is a covalent bond. In some embodiments, L P3 is an optionally substituted divalent C2-C6 aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L P3is 0 to 10 atoms in length. P3 is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P3 is or comprises an amino acid residue. P3 In some embodiments, L is or comprises a peptide. P3 is -[X]pX 6 X 7 [X]p'-, and each of X, X 6 , and X 7 are independently an amino acid residue, and each of p and p' is independently 0 to 10. P3 -X 6 X 7 - is or contains each X 6 and X 7 are independently amino acid residues. 7 L AA3 In some embodiments, L P3In some embodiments, the methylene unit of X is replaced with -C(R')2-, and one of the R' groups is the fifth, sixth, seventh, or eighth R' group. 7 comprises -C(R')2-, where one of the R' groups is the fifth, sixth, seventh, or eighth R' group. L AA3

[0280] In some embodiments, L AA3 is or comprises an amino acid residue. AA3 is or comprises an amino acid residue comprising a side chain containing an acidic or polar group. AA3 is an amino acid residue containing a side chain containing an acidic group.

[0281] In some embodiments, L AA3 L AR and the methylene unit is -C(R')(R AS )-, where each variable is independently as described herein. AA3 is an optionally substituted divalent C1-C6 (e.g., C1, C2, C3, C4, C5, or C6) aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA3 is an optionally substituted divalent C2-C4 aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R ASIn some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA3 is -N(R')-C(R')(R AS )—C(O)—, and each variable is independently as described herein. In some embodiments, L AA3 is -NH-C(R')(R AS )—C(O)—, and each variable is independently as described herein.

[0282] In some embodiments, L AS3 is, as described herein, L AS In some embodiments, R AA3 is —COR, where R is as described herein. In some embodiments, R is H. In some embodiments, L AA3 is an acidic amino acid residue, e.g., a residue such as Asp, Glu, etc. In some embodiments, L AA3 is, as described herein, X 6 is.

[0283] In some embodiments, L AA3 In some embodiments, L AA3 is or comprises a hydrophobic amino acid residue. AA3 is X as described herein. 8 is. L P4

[0284] In some embodiments, L P4 is a covalent bond or an optionally substituted divalent is a C2-C6 valent aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. P4 is 0 to 10 atoms in length. P4 is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P4 is or comprises an amino acid residue. P4 is or comprises a peptide.

[0285] In some embodiments, L P4 is -[X]pX 7 X 8 [X]p'- or containing it, and each of X, X 7 and X8 are independently an amino acid residue, and each of p and p' is independently 0 to 10. P4 -X 7 X 8 - is or contains each X 7 and X 8 are independently as described herein; and X 8 L AA4 is bonded to.

[0286] In some embodiments, L P4 In some embodiments, the methylene unit of X is replaced with -C(R')2-, and one of the R' groups is the fifth, sixth, seventh, or eighth R' group. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is 7 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 7 In some embodiments, it is the alpha carbon of R'. In some embodiments, it is the fifth R' group. In some embodiments, it is the sixth R' group. In some embodiments, it is the seventh R' group. In some embodiments, it is the eighth R' group. L AA4

[0287] In some embodiments, L AA4 is or comprises an amino acid residue. AA4 is or comprises an amino acid residue that includes a side chain that includes an aromatic group.

[0288] In some embodiments, L AA4 L AR and the methylene unit is -C(R')(R AS )-, where each variable is independently as described herein. AA4is an optionally substituted divalent C1-C6 (e.g., C1, C2, C3, C4, C5, or C6) aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA4 is an optionally substituted divalent C2-C4 aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA4 is -N(R')-C(R')(R AS )—C(O)—, and each variable is independently as described herein. In some embodiments, L AA4 is -NH-C(R')(R AS )—C(O)—, and each variable is independently as described herein.

[0289] In some embodiments, L AS4 is, as described herein, L AS In some embodiments, R AA4 is replaced by C 6~14 In some embodiments, R AA4 is optionally substituted phenyl. In some embodiments, R AA4is phenyl. In some embodiments, R AA4 is an optionally substituted 10-membered C 10 In some embodiments, R AA4 is an optionally substituted 5-membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 is an optionally substituted 6-membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 is an optionally substituted 9-membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA4 is an optionally substituted 10-membered bicyclic heteroaryl having 1-4 heteroatoms. In some embodiments, the heteroaryl has no more than one heteroatom. In some embodiments, the heteroaryl has two or more heteroatoms. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is sulfur. In some embodiments, R AA4 is replaced as necessary [ka] In some embodiments, R AA4 is replaced as necessary [ka] In some embodiments, R AA4 is replaced as necessary [ka] In some embodiments, L AA4 is an aromatic amino acid residue, as described herein. AA4 is, as described herein, X 9 is. L P5

[0290] In some embodiments, L P5 is a covalent bond or an optionally substituted divalent C-C aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L P5 is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P5 is or comprises an amino acid residue. P5 is or comprises a peptide.

[0291] In some embodiments, L P5 is -[X]pX 11[X]p'-, where each variable is independently as described herein. P5 -X 10 X 11 - is or contains each X 10 and X 11 are independently as described herein; and X 11 L AA5 is bonded to.

[0292] In some embodiments, L P5 is a -C(R')2- group, and one of the R' groups is a fourth R' group. P5 is a -C(R')2- group, and one of the R' groups is a second R' group. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of an amino acid residue. 11 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 11 is the alpha carbon of

[0293] In some embodiments, L P5 contains a -C(R')2- group, where one of the R' groups is the fifth, sixth, seventh, or eighth R' group. In some embodiments, such a -C(R')2- group is that of an amino acid residue. In some embodiments, such a -C(R')2- group is that of an X 10 In some embodiments, such a carbon atom is the alpha carbon of an amino acid residue. In some embodiments, such a carbon atom is X 10 In some embodiments, it is the alpha carbon of R'. In some embodiments, it is the fifth R' group. In some embodiments, it is the sixth R' group. In some embodiments, it is the seventh R' group. In some embodiments, it is the eighth R' group. L AA5

[0294] In some embodiments, LAA5 is or comprises an amino acid residue. AA5 is or comprises an amino acid residue that includes a side chain that includes an aromatic group.

[0295] In some embodiments, L AA5 L AR and the methylene unit is -C(R')(R AS )-, where each variable is independently as described herein. AA5 is an optionally substituted divalent C1-C6 (e.g., C1, C2, C3, C4, C5, or C6) aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA5 is an optionally substituted divalent C2-C4 aliphatic group, wherein one or more methylene units of the group are optionally independently selected from -C(R')2-, -C(R')(R AS In some embodiments, L is replaced with -Cy-, -O-, -S-, -SS-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)N(R')-, -C(O)S-, or -C(O)O-, wherein each variable is independently as described herein. AA5 is -N(R')-C(R')(R AS )—C(O)—, and each variable is independently as described herein. In some embodiments, L AA5 is -NH-C(R')(R AS)—C(O)—, and each variable is independently as described herein.

[0296] In some embodiments, L AS5 is, as described herein, L AS In some embodiments, R AA5 is replaced by C 6~14 In some embodiments, R AA5 is optionally substituted phenyl. In some embodiments, R AA5 is phenyl. In some embodiments, R AA5 is an optionally substituted 10-membered C 10 In some embodiments, R AA5 is an optionally substituted 5-membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 is an optionally substituted 6-membered monocyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 is an optionally substituted 9-membered bicyclic heteroaryl having 1 to 4 heteroatoms. In some embodiments, R AA5 is an optionally substituted 10-membered bicyclic heteroaryl having 1-4 heteroatoms. In some embodiments, the heteroaryl has no more than one heteroatom. In some embodiments, the heteroaryl has two or more heteroatoms. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is sulfur. In some embodiments, R AA5 is replaced as necessary [ka] In some embodiments, R AA5 is replaced as necessary [ka] In some embodiments, RAA5 is replaced as necessary [ka] In some embodiments, L AA5 is an aromatic amino acid residue, as described herein. AA5 is, as described herein, X 12 is. L P6

[0297] In some embodiments, L P6 is a covalent bond. In some embodiments, L P6 is an optionally substituted divalent C2-C6 aliphatic group, wherein one or more methylene units of the group are optionally independently replaced with -C(R')2-, -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -C(O)S-, or -C(O)O-. In some embodiments, L P6 is 0 to 10 atoms in length (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, etc.). P6is 2 to 10 atoms in length. In some embodiments, it is 2 atoms. In some embodiments, it is 3 atoms. In some embodiments, it is 4 atoms. In some embodiments, it is 5 atoms. In some embodiments, it is 6 atoms. In some embodiments, it is 7 atoms. In some embodiments, it is 8 atoms. In some embodiments, it is 9 atoms. In some embodiments, it is 10 atoms. In some embodiments, one or more methylene units are independently replaced with -N(R')-, -C(R')2-, -C(O)-, or -C(O)N(R')-. In some embodiments, a methylene unit is replaced with -N(R')-. In some embodiments, a methylene unit is replaced with -C(R')2-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)-. In some embodiments, a methylene unit is replaced with -C(O)N(R')-. In some embodiments, each methylene unit is independently replaced with -N(R')-, -C(R')2-, or -C(O)-. P6 is or comprises an amino acid residue. P6 is or comprises a peptide. L AA6

[0298] In some embodiments, L AA6 is or comprises an amino acid residue. AA6 is or comprises an amino acid residue that includes a side chain that includes an aromatic group.

[0299] In some embodiments, L AA6 ...

Claims

1. as follows: 【Chemical Formula 362-1】 【Chemical Formula 362-2】 【Chemical Formula 362-3】 【Chemical Formula 362-4】 【Chemical Formula 362-5】 a drug having the structure or a salt thereof.

2. A drug, wherein the drug is [X] ｐ X １ X ２ X ３ X ４ X ５ X ６ X ７ X ８ X ９ X １０ X １１ X １２ X １３ X １４ [X １５ ] ｐ１５ [X １６ ] ｐ１６ [X １７ ] ｐ１７ [X] ｐ’ or includes it, wherein each of p15, p16, and p17 is independently 0 or 1, each of p and p' is independently 0 to 10, X, X １ , X ２ , X ３ , X ４ , X ５ , X ６ , X ７ , X ８ , X ９ , X １０ , X １１ , X １２ , X １３ , X １４ , X １５ , X １６ , and X １７ each of which is independently an amino acid residue, agent. **Claim 3** The agent is [X ０ ｐ０ X １ X ２ X ３ X ４ X ５ X ６ X ７ X ８ X ９ X １０ X １１ X １２ X １３ X １４ [X １５ ｐ１５ [X １６ ｐ１６ [X １７ ｐ１７ a peptide containing it, or containing it, wherein each of p0, p15, p16, and p17 is independently 0 or 1, X ０ , X １ , X ２ , X ３ , X ４ , X ５ , X ６ , X ７ , X ８ , X ９ , X １０ , X １１ , X １２ , X １３ , X １４ , X １５ , X １６ , and X １７ each of which is independently an amino acid residue, X ２ contains a side chain containing an acidic group or a polar group, X ５ contains a side chain containing an acidic group or a polar group, X ９ , X １２ , and X １３each of which contains a side chain containing a substituted or unsubstituted aromatic group The agent according to claim 2.

4. The agent according to any one of claims 2 to 3, wherein the agent contains three or more staples within 10 to 20 amino acid residues.

5. X ０ , X １ , X ３ , X ４ , X ７ , X １０ , X １１ , and X １４ five of which are each independently an amino acid residue for stapling or are each independently stapled, and optionally, X1 and X4 are connected by a staple. X0 and X4 are connected by a staple. X4 and X11 are connected by a staple. X10 and X14 are connected by a staple. X7 and X10 are connected by a staple and / or, X7 and X14 are connected by a staple. The agent according to any one of claims 2 to 4.

6. The agent according to any one of claims 2 to 5, wherein the agent contains an N-terminal group.

7. X １ is a residue of an amino acid having a structure of formula A-I, A-II, or A-III, and R ａ１ and R ａ３ together with their intervening atoms form a 3- to 10-membered ring having 0 to 5 heteroatoms in addition to the intervening atoms, which is substituted or unsubstituted. The agent according to any one of claims 2 to 6.

8. X ４ is a residue of an amino acid containing an olefin, the agent according to any one of claims 2 to 7.

9. X １０ is a residue of an amino acid containing a substituted or unsubstituted carboxyl group, a substituted or unsubstituted amino group, an azidyl group, a substituted or unsubstituted alkynyl group, or a substituted or unsubstituted thiol group, the agent according to any one of claims 2 to 8.

10. X １１ is a residue of an amino acid containing an olefin, the agent according to any one of claims 2 to 9.

11. X １４ is a residue of an amino acid containing a carboxyl group, an amino group, an azidyl group, an alkynyl group, or a thiol group, the agent according to any one of claims 2 to 10.

12. X １０ and X １４ one of which is a residue of an amino acid containing a carboxyl group and the other is a residue of an amino acid containing an amino group, the agent according to any one of claims 2 to 11.

13. X １０ and X １４ are connected by a staple, the staple containing -C(O)N(R')-, the agent according to any one of claims 2 to 12.

14. X ２ contains a side chain containing an acidic group, the agent according to any one of claims 2 to 13.

15. X ３ contains one or two hydrophobic side chains, the agent according to any one of claims 2 to 14.

16. X ５The agent according to any one of claims 2 to 15, which comprises a side chain containing an acidic group.

17. X ６ The agent according to any one of claims 2 to 16, which comprises a side chain containing an acidic group.

18. X ７ The agent according to any one of claims 2 to 17, wherein X is a hydrophobic amino acid residue.

19. X ８ The agent according to any one of claims 2 to 18, wherein X is a hydrophobic amino acid residue.

20. X ９ The agent according to any one of claims 2 or 4 to 19, which comprises a side chain containing a substituted or unsubstituted aromatic group.

21. X １２ The agent according to any one of claims 2 or 4 to 20, which comprises a side chain containing a substituted or unsubstituted aromatic group.

22. X13 is the agent according to any one of claims 2 or 4 to 21, which comprises a side chain containing a substituted or unsubstituted aromatic group.

23. p15 is 1, and optionally, X15 is the agent according to any one of claims 2 to 22, which comprises a hydrophobic side chain.

24. The peptide has a sequence of SEQ ID NO: 2 or a fragment thereof: SVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKMVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTSRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEGLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRT (SEQ ID NO: 2) binds to a polypeptide that is or contains it, or the peptide binds to beta-catenin and at least two, or at least three, or at least four, or at least five, or at least six, or at least seven, or at least eight, or at least nine, or at least ten, or at least eleven, or at least twelve, or at least thirteen, or at least fourteen, or at least fifteen, or at least sixteen, or at least seventeen, or at least eighteen, or at least nineteen, or at least twenty of the following amino acid residues at the positions shown in SEQ ID NO: 1: A305, Y306, G307, N308, Q309, K312, R342, K345, V346, V349, Q375, R376, Q379, N380, L382, W383, R386, N387, D413, N415, V416, T418, and C419, or interacts with one or more residues corresponding thereto, the agent according to any one of claims 2 to 23.

25. the double bond of the (i, i + 7) staple is E, the agent according to any one of claims 1 to 24.

26. the double bond of the (i, i + 7) staple is Z, the agent according to any one of claims 1 to 24.

27. the double bond of the (i, i + 2), (i, i + 3), or (i, i + 4) staple is E, the agent according to any one of claims 1 to 26.

28. the double bond of the (i, i + 2), (i, i + 3), or (i, i + 4) staple is Z, the agent according to any one of claims 1 to 26.

29. the carbon atoms bonded to two staples are of the R configuration, the agent according to any one of claims 1 to 28.

30. A drug having the structure of SP-1-1, SP-1-2, SP-1-3, SP-1-4, SP-1-5, SP-1-6, SP-1-7, SP-1-8, SP-2-1, SP-2-2, SP-2-3, SP-2-4, SP-2-5, SP-2-6, SP-2-7, SP-2-8, SP-3-1, SP-3-2, SP-4-1, SP-4-2, SP-4-3, SP-4-4, SP-4-5, SP-4-6, SP-4-7, SP-4-8, SP-5-1, SP-5-2, SP-5-3, SP-5-4, SP-5-5, SP-5-6, SP-5-7, SP-5-8, SP-6, SP-7-1, SP-7-2, SP-7-3, SP-7-4, SP-7-5, SP-7-6, SP-7-7, SP-7-8, SP-8-1, SP-8-2, SP-8-3, SP-8-4, SP-8-5, SP-8-6, SP-8-7, SP-8-8, SP-9-1, SP-9-2, SP-9-3, SP-9-4, SP-9-5, SP-9-6, SP-9-7, SP-9-8, SP-10-1, SP-10-2, SP-10-3, SP-10-4, SP-10-5, SP-10-6, SP-10-7, SP-10-8, SP-11-1, SP-11-2, SP-11-3, SP-11-4, SP-11-5, SP-11-6, SP-11-7, SP-11-8, SP-12-1, SP-12-2, SP-12-3, SP-12-4, SP-12-5, SP-12-6, SP-12-7, SP-12-8, SP-13-1, SP-13-2, SP-13-3, SP-13-4, SP-13-5, SP-13-6, SP-13-7, SP-13-8, SP-14-1, SP-14-2, SP-14-3, SP-14-4, SP-14-5, SP-14-6, SP-14-7, SP-14-8, SP-15-1, SP-15-2, SP-15-3, SP-15-4, SP-15-5, SP-15-6, SP-15-7, SP-15-8, or a salt thereof.

31. as follows: 【Chemical Formula 363】 A drug having the structure of, or a salt thereof, or as follows: 【Chemical Formula 364】 A drug having the structure of, or a salt thereof. Claim 32. The agent has the same retention time as I-66 prepared as described in Example 9 under HPLC conditions, and the HPLC conditions can separate I-66 and I-67 prepared as described in Example 9, or, The agent exhibits a retention time of 15.3 minutes under the following HPLC conditions: Agilent Poroshell 120 EC-C18; 4.6×100 mm; Solvent A = 0.1% TFA in water; Solvent B = 0.075% TFA in acetonitrile; The gradient is from 10% B to 95% B over 30 minutes; Detection is UV absorption at 220 nm. The agent according to claim 31.

33. The agent elutes as a single peak having I-66 prepared as described in Example 9 under the following HPLC conditions: Agilent Poroshell 120 EC-C18; 4.6×100 mm; Solvent A = 0.1% TFA in water; Solvent B = 0.075% TFA in acetonitrile; The gradient is from 10% B to 95% B over 30 minutes; Detection is UV absorption at 220 nm, or, The agent exhibits 1H NMR peaks that overlap with the peaks at 5.1 to 5.7 in Figure 6 under the same or equivalent conditions, or, The agent exhibits the same 1H NMR peaks as those at 5.1 to 5.7 in Figure 6 under the same or equivalent conditions, or, In its 1H NMR spectrum, the peaks corresponding to 1H bonded to carbon atoms overlap with the peaks in Figure 6 under the same or equivalent conditions, or, Its 1H NMR spectrum overlaps with the peaks in Figure 6 under the same or equivalent conditions. The agent according to any one of claims 31 to 32.

34. The drug has the same retention time as I-67 prepared as described in Example 9 under HPLC conditions, and the HPLC conditions can separate I-66 and I-67 prepared as described in Example 9, or, The drug shows a retention time of 16.2 minutes under the following HPLC conditions: Agilent Poroshell 120 EC-C18; 4.6×100 mm; Solvent A = 0.1% TFA in water; Solvent B = 0.075% TFA in acetonitrile; the gradient is from 10% B to 95% B over 30 minutes; the detection is UV absorption at 220 nm, or, The drug elutes as a single peak having I-67 prepared as described in Example 9 under the following HPLC conditions: Agilent Poroshell 120 EC-C18; 4.6×100 mm; Solvent A = 0.1% TFA in water; Solvent B = 0.075% TFA in acetonitrile; the gradient is from 10% B to 95% B over 30 minutes; the detection is UV absorption at 220 nm, or, The drug shows 1H NMR peaks that do not overlap with the peaks at 5.1 - 5.7 in Figure 6 under the same or equivalent conditions, or, The drug is characterized by not showing the same 1H NMR peaks as those at 5.1 - 5.7 in Figure 6 under the same or equivalent conditions, or, In its 1H NMR spectrum, the peaks corresponding to 1H bonded to carbon atoms do not all overlap with the peaks in Figure 6 under the same or equivalent conditions, or, The 1H NMR spectrum of the drug does not overlap with the peaks in Figure 6 under the same or equivalent conditions. The drug according to claim 31.

35. The carbon atoms bonded to the two staples have an S configuration. The drug according to any one of claims 1 to 34.

36. The structure of formula I R Ｎ -L Ｐ１ -L ＡＡ１-L Ｐ２ -L ＡＡ２ -L Ｐ３ -L ＡＡ３ -L Ｐ４ -L ＡＡ４ -L Ｐ５ -L ＡＡ５ -L Ｐ６ -L ＡＡ６ -L Ｐ７ -R Ｃ I a drug having the same or a salt thereof, wherein in the formula R Ｎ is a peptide, an amino protecting group, or R'-L ＲＮ - and L Ｐ１ L Ｐ２ L Ｐ３ L Ｐ４ L Ｐ５ L Ｐ６ and L Ｐ７ each is independently L, and L Ｐ１ L Ｐ２ L Ｐ３ L Ｐ４ L Ｐ５ L Ｐ６ and L Ｐ７ are the first and second R' groups that together form -L ｓ - where -L ｓ - is bonded to the atoms to which the first R' group and the second R' group are bonded, the first R' group and the second R' group, and the third and fourth R' groups that together form -L ｓ - where -L ｓ - is bonded to the atoms to which the third R' group and the fourth R' group are bonded, the third R' group and the fourth R' group are included, each L ｓ is independently -L ｓ１ -L ｓ２ -L ｓ３ - and each L ｓ１ L ｓ２ and L ｓ３ are independently L, L ＡＡ１ is an amino acid residue containing a side chain having an acidic group or a polar group, L ＡＡ２ is an amino acid residue containing a side chain having an acidic group or a polar group, L ＡＡ３ is an amino acid residue, L ＡＡ４ is an amino acid residue containing a side chain having a substituted or unsubstituted aromatic group, L ＡＡ５ is an amino acid residue containing a side chain having a substituted or unsubstituted aromatic group, L ＡＡ６ is an amino acid residue containing a side chain having a substituted or unsubstituted aromatic group, R Ｃ is a peptide, a carboxyl protecting group, -L ＲＣ -R', -O-L ＲＣ -R', or -N(R')-L ＲＣ -R', L ＲＮ and L ＲＣ each of which is independently L, each L is independently a covalent bond or a substituted or unsubstituted divalent C １ ~C ２５ aliphatic group or a divalent C having 1 to 10 heteroatoms １ ~C ２５ heteroaliphatic group, wherein one or more methylene units of said group are independently -C(R') ２ -, -Cy-, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ N(R')-, -C(O)S-, or -C(O)O- and are either replaced or not replaced, Each -Cy- is, independently, a substituted or unsubstituted divalent 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms, Each R' is, independently, -L-R, -C(O)R, -COR, ２ R, or -SO ２ R, Each R is, independently, -H or a substituted or unsubstituted group selected from C １～３０ aliphatic, C having 1 to 10 heteroatoms １～３０ heteroaliphatic, C ６～３０ aryl, C ６～３０ arylaliphatic, C having 1 to 10 heteroatoms ６～３０ arylheteroaliphatic, 5- to 30-membered heteroaryl having 1 to 10 heteroatoms, and 3- to 30-membered heterocyclyl having 1 to 10 heteroatoms, or two R groups, independently, together form a covalent bond or do not together form said covalent bond, or two or more R groups on the same atom, independently, together with said atom form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said atom or do not together with said atom form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, or two or more R groups on two or more atoms, independently, together with their intervening atoms form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said intervening atoms or do not together with said intervening atoms form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, a drug or a salt thereof, or the structure of formula I R Ｎ -L Ｐ１ -L ＡＡ１ -L Ｐ２ -L ＡＡ２ -L Ｐ３-L ＡＡ３ -L Ｐ４ -L ＡＡ４ -L Ｐ５ -L ＡＡ５ -L Ｐ６ -L ＡＡ６ -L Ｐ７ -R Ｃ I which is a drug having I or a salt thereof, wherein, R Ｎ is a peptide, an amino protecting group, or R'-L ＲＮ - and, L Ｐ１ L, Ｐ２ L, Ｐ３ L, Ｐ４ L, Ｐ５ L, Ｐ６ and L Ｐ７ each independently is L, and L Ｐ１ L, Ｐ２ L, Ｐ３ L, Ｐ４ L, Ｐ５ L, Ｐ６ and L Ｐ７ are, together a first R' group and a second R' group forming -L ｓ -, where -L ｓ - is the first R' group and the second R' group that bind to the atoms to which the first R' group binds and the second R' group binds, and together a third R' group and a fourth R' group forming -L ｓ -, where -L ｓ - is the third R' group and the fourth R' group that bind to the atoms to which the third R' group binds and the fourth R' group binds and contain, each L ｓ independently is -L ｓ１ -L ｓ２ -L ｓ３ -, and each L ｓ１ L, ｓ２ and L ｓ３ independently are L, L ＡＡ１ is L ＡＲand the methylene unit is replaced by -C(R')(R ＡＳ ), where R ＡＳ is -L ＡＳ１ -R ＡＡ１ , and R ＡＡ１ is -COR or -SO ２ R, and ２ L ＡＡ２ is L ＡＲ and the methylene unit is replaced by -C(R')(R ＡＳ ), where R ＡＳ is -L ＡＳ２ -R ＡＡ２ , and R ＡＡ２ is -COR or -SO ２ R, and ２ L ＡＡ３ is L ＡＲ and the methylene unit is replaced by -C(R')(R ＡＳ ), where R ＡＳ is -L ＡＳ３ -R ＡＡ３ , and R ＡＡ３ is R', and L ＡＡ４ is L ＡＲ and the methylene unit is replaced by -C(R')(R ＡＳ ), where R ＡＳ is -L ＡＳ４ -R ＡＡ４ , and R ＡＡ４ is a substituted or unsubstituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms, L ＡＡ５ is L ＡＲ and the methylene unit is replaced by -C(R')(R ＡＳ ), where R ＡＳ is -L ＡＳ５ -R ＡＡ５ , and R ＡＡ５ is a substituted or unsubstituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms, L ＡＡ６ is L ＡＲ ​​and the methylene unit is replaced by -C(R')(R ＡＳ ), R ＡＳ is -L ＡＳ６ -R ＡＡ６ where R ＡＡ６ is a substituted or unsubstituted group selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl having 1 to 6 heteroatoms, R Ｃ is a peptide, a carboxyl protecting group, -L ＲＣ -R', -O-L ＲＣ -R', or -N(R')-L ＲＣ -R', L ＲＮ and each of L ＲＣ is independently L, each L ＡＲ is independently a substituted or unsubstituted divalent C １ to C ６ aliphatic group, one or more methylene units of said group being independently -C(R') ２ -, -C(R')(R ＡＳ ), -Cy-, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ (-), -S(O) ２ N(R')-, -C(O)S-, or -C(O)O- replaced or not replaced, L ＡＳ１ L ＡＳ２ L ＡＳ３ L ＡＳ４ L ＡＳ５ and each of L ＡＳ６ is independently L ＡＳ (-), each R ＡＳ is independently -L ＡＳ -R', each L ＡＳ is independently a covalent bond or a substituted or unsubstituted divalent C １ to C １０ an aliphatic group or a divalent C having 1 to 5 heteroatoms １ ~C １０ heteroaliphatic group, wherein one or more methylene units of said group are independently -C(R') ２ -, -Cy-, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ N(R')-, -C(O)S-, or -C(O)O- or not replaced, each L is independently a covalent bond or a substituted or unsubstituted divalent C １ ~C ２５ aliphatic group or a divalent C having 1 to 10 heteroatoms １ ~C ２５ heteroaliphatic group, wherein one or more methylene units of said group are independently -C(R') ２ -, -Cy-, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ N(R')-, -C(O)S-, or -C(O)O- or not replaced, each -Cy- is independently a substituted or unsubstituted divalent 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms, each R' is independently -L-R, -C(O)R, -COR ２ R, or -SO ２ R, each R is independently -H or C １～３０ aliphatic, C having 1 to 10 heteroatoms １～３０ heteroaliphatic, C ６～３０ aryl, C ６～３０Aryl aliphatic, C having 1 to 10 heteroatoms ６～３０ A substituted or unsubstituted group selected from aryl heteroaliphatic, 5- to 30-membered heteroaryl having 1 to 10 heteroatoms, and 3- to 30-membered heterocyclyl having 1 to 10 heteroatoms, or two R groups, independently, may together form a covalent bond or may not together form said covalent bond, or two or more R groups on the same atom, independently, together with said atom, form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said atom or do not together with said atom form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, or two or more R groups on two or more atoms, independently, together with their intervening atoms, form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said intervening atoms or do not together with said intervening atoms form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, a drug or a salt thereof.

37. A drug according to any one of claims 1 to 26, wherein each olefin double bond in the staple is independently converted to a single bond or not converted to said single bond.

38. A drug according to any one of claims 1 to 27, having a diastereomeric purity of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher, or having a purity of 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher.

39. A pharmaceutical composition comprising or delivering a drug according to any one of claims 1 to 38 and a pharmaceutically acceptable carrier.

40. A composition selected from Table E2 or Table E3, Optionally, the composition comprises or delivers one or more or all of the peptide agents in the composition selected from Table E2 or Table E3 and a pharmaceutically acceptable carrier, A pharmaceutical composition characterized by being a composition.

41. A drug comprising one or more staples each independently containing one or more olefin double bonds, wherein the ratio of the two stereoisomers of the olefin double bond in the staple is 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 30:1, 40:1, 50:1 or greater, The composition according to any one of claims 39 to 40.

42. a) preparing a first compound comprising two moieties each independently containing an olefin double bond; b) providing a second compound by stapling said two moieties by olefin metathesis of the olefin double bond of one moiety with the olefin double bond of the other moiety to form a first formed staple; c) adding one or more additional moieties to said second compound to provide a third compound each independently comprising two moieties containing an olefin double bond, and d) providing a fourth compound by stapling said two moieties in said third compound by olefin metathesis of the olefin double bond of one moiety with the olefin double bond of the other moiety to form a second formed staple A method comprising.

43. A method of modulating the interaction of beta-catenin with a partner in a system, the method comprising contacting beta-catenin with the composition, or A method of modulating the interaction of beta-catenin with its partners in a system, said method comprising administering or delivering said composition to said system, or A method of modulating TCF-beta-catenin interaction in a system, said method comprising contacting beta-catenin with said composition, or A method of modulating TCF-beta-catenin interaction in a system, said method comprising administering or delivering said composition to said system, or A method of inhibiting beta-catenin-dependent cell proliferation, said method comprising administering or delivering said composition to said system, or A method of modulating the WNT / beta-catenin pathway in a system, said method comprising administering or delivering said composition to said system, wherein the expression of a nucleic acid is modulated, or A method comprising administering or delivering said composition to said system, wherein the level of a nucleic acid transcript and / or its product is modulated, or A method comprising administering or delivering said composition to said system, wherein the expression of a nucleic acid is modulated A composition for use in, comprising a medicament according to any one of claims 1 to 38, or a composition according to any one of claims 39 to 41.

44. A method of treating or preventing a condition, disorder, or disease associated with the interaction of beta-catenin with its partners in a subject, said method comprising administering or delivering said composition to said subject, wherein optionally, said partner is TCF7, LEF1, TCF7L1, or TCF7L2 A composition for use in, comprising a medicament according to any one of claims 1 to 38, or a composition according to any one of claims 39 to 41.

45. A method of treating cancer in a subject, the method comprising administering or delivering the composition to the subject A composition for use in , comprising a drug according to any one of claims 1 to 38, or a composition according to any one of claims 39 to 41. **Claim 46** The method comprises administering or delivering a second therapeutic agent or therapy to the subject, optionally, the second therapeutic agent is a chemotherapeutic agent, a hormonal therapy agent, an immunotherapy agent, a checkpoint inhibitor, an antibody, a CTLA-4, a PD-1, or a PD-L1 inhibitor, or a cell, or comprises the same, or the second therapy is surgery, chemotherapy, radiotherapy, hormonal therapy, stem cell or bone marrow transplantation, immunotherapy, T cell therapy, or CAR T cell therapy, or comprises the same, optionally, the method further comprises evaluating the expression of nucleic acids. A composition for use according to any one of claims 43 to 45. **Claim 47** The structure of formula PA N(R ＰＡ )(R ａ１ )-L ａ１ -C(R ａ２ )(R ａ３ )-L ａ２ -C(O)R ＰＣ PA A compound having the formula or a salt thereof, wherein, R ＰＡ is -H or an amino protecting group, R ａ１ and each of R ａ３ is independently -L ａ -R', R ａ２ is -L ａａ -C(O)R ＰＳ wherein, L ａ , L ａ１ , and each of L ａ２ is independently L, -C(O)R ＰＳ is -COOH which is protected or activated as necessary, -C(O)R ＰＣ is -COOH which is protected or activated as necessary, Each L is independently a covalent bond or a substituted or unsubstituted divalent C １ ~C ２５ aliphatic group or a divalent C having 1 to 10 heteroatoms １ ~C ２５ heteroaliphatic group, and one or more methylene units of said group are independently -C(R') ２ -, -Cy-, -O-, -S-, -S-S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O) ２ -, -S(O) ２ N(R')-, -C(O)S-, or -C(O)O- is replaced or not replaced, Each -Cy- is independently a substituted or unsubstituted divalent 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms, Each R' is independently -R, -C(O)R, -CO ２ R, or -SO ２ R, Each R is independently -H or C １～３０ aliphatic, C having 1 to 10 heteroatoms １～３０ heteroaliphatic, C ６～３０ aryl, C ６～３０ arylaliphatic, C having 1 to 10 heteroatoms ６～３０ arylheteroaliphatic, 5- to 30-membered heteroaryl having 1 to 10 heteroatoms, and 3- to 30-membered heterocyclyl having 1 to 10 heteroatoms, which is a substituted or unsubstituted group selected from, or The two R groups are independently either joined together to form a covalent bond or not joined together to form said covalent bond, or two or more R groups on the same atom are independently joined with said atom to form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said atom, or not joined with said atom to form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, or two or more R groups on two or more atoms are independently joined with their intervening atoms to form a substituted or unsubstituted 3- to 30-membered monocyclic, bicyclic, or polycyclic ring having 0 to 10 heteroatoms in addition to said intervening atoms, or not joined with said intervening atoms to form said 3- to 30-membered monocyclic, bicyclic, or polycyclic ring, a compound or a salt thereof, or the following structure 【Chemical Formula 365】 a compound having the same or a salt thereof, wherein in the formula, R ＰＡ is -H or an amino protecting group, -C(O)R ＰＳ is -COOH which is protected or activated as necessary, -C(O)R ＰＣ is -COOH which is protected or activated as necessary, a compound or a salt thereof, or the following structure 【Chemical Formula 366】 a compound having the same or a salt thereof, wherein in the formula, R ＰＡ is -H or an amino protecting group, -C(O)R ＰＳ is -COOH which is protected or activated as necessary, -C(O)R ＰＣwhich is -COOH protected or activated as necessary, a compound or a salt thereof.

48. A method for modulating the interaction of beta-catenin with its partner in a system, the method comprising the step of contacting beta-catenin with the composition, or A method for modulating the interaction of beta-catenin with its partner in a system, the method comprising the step of administering or delivering the composition to the system, or A method for modulating the TCF-beta-catenin interaction in a system, the method comprising the step of contacting beta-catenin with the composition, or A method for modulating the TCF-beta-catenin interaction in a system, the method comprising the step of administering or delivering the composition to the system, or A method for inhibiting beta-catenin-dependent cell proliferation, the method comprising the step of administering or delivering the composition to the system, or A method for modulating the WNT / beta-catenin pathway in a system, the method comprising the step of administering or delivering the composition to the system, wherein the expression of a nucleic acid is modulated, or A method comprising the step of administering or delivering the composition to the system, wherein the level of a nucleic acid transcript and / or its product is modulated, or A method comprising the step of administering or delivering the composition to the system, wherein the expression of a nucleic acid is modulated The use of a composition comprising the agent according to any one of claims 1 to 38, or the use of the composition according to any one of claims 39 to 41, in the manufacture of a medicament for use in.

49. A method for treating or preventing a condition, disorder, or disease associated with the interaction of beta-catenin with a partner in a subject, the method comprising administering or delivering the composition to the subject, wherein, optionally, the partner is TCF7, LEF1, TCF7L1, or TCF7L2 Use in the manufacture of a medicament for use in, a composition comprising a medicament according to any one of claims 1 to 38, or use of a composition according to any one of claims 39 to 41.

50. A method for treating cancer in a subject, the method comprising administering or delivering the composition to the subject Use in the manufacture of a medicament for use in, a composition comprising a medicament according to any one of claims 1 to 38, or use of a composition according to any one of claims 39 to 41.

51. The method comprising administering or delivering a second therapeutic agent or treatment to the subject, wherein, optionally, the second therapeutic agent is or comprises a chemotherapeutic agent, a hormonal therapy agent, an immunotherapeutic agent, a checkpoint inhibitor, an antibody, a CTLA-4, PD-1, or PD-L1 inhibitor, or a cell, or the second treatment is or comprises surgery, chemotherapy, radiotherapy, hormonal therapy, stem cell or bone marrow transplantation, immunotherapy, T cell therapy, or CAR T cell therapy, wherein, optionally, the method further comprises assessing the expression of a nucleic acid Use according to any one of claims 48 to 50.

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