Peptide conjugates of microtubule targeting agents as therapeutic agents

Abstract

The present invention relates to peptide conjugates of microtubule targeting agents, such as maytansine derivatives, which are useful in treating diseases such as cancer.

Description

Technical Field

[0001] The present invention relates to peptide conjugates of microtubule targeting agents, such as maytansinoid derivatives, which are useful in treating diseases such as cancer. Background of the Invention

[0003] Cancer is a group of diseases characterized by abnormal control of cell growth. In the United States alone, the annual incidence of cancer is estimated to exceed 1.6 million. Despite the use of surgery, radiation, chemotherapy, and hormone therapy to treat cancer, it remains the second leading cause of death in the United States. An estimated 600,000 Americans will die from cancer each year.

[0004] Cancer treatment in humans by systemic administration of pharmaceutical agents typically works by slowing or stopping the uncontrolled replication that is a characteristic of cancer cells. One such agent is a microtubule-targeting agent. Cell division requires the formation of a complete mitotic spindle, which is composed of microtubules that undergo random length variations. The random length variations of microtubules are known as dynamic instability. Disrupting the dynamic instability of microtubules can lead to inhibition of further cell division. Drugs that target microtubules to inhibit dynamic instability are currently used in the clinic as effective anticancer agents for a wide variety of cancers. See Lopus, M, Cancer Lett., 2011, 307(2): 113-118.

[0005] Maytansines (e.g., mertansine, DM1 or DM4) are a class of microtubule targeting agents that have emerged as potential clinical chemotherapeutic agents. See Lopus, M, Cancer Lett., 2011, 307(2): 113-118; and Widdison, W., J. Med. Chem. 2006, 49: 4392-4408. Although DM1 has been shown to be effective in treating several types of cancer (including lymphoma and breast cancer), the toxic side effects of, for example, peripheral neuropathy hinder the clinical development of microtubule targeting agents such as maytansines. Preferential delivery of maytansine compounds (e.g., DM1) to diseased tissues can avoid these serious side effects. Therefore, it is necessary to more selectively deliver maytansine compounds to diseased tissues. Summary of the Invention

[0006] The present disclosure provides, inter alia, a compound of formula (I):

[0007] R 2 -LR 1 (I)

[0008] or a pharmaceutically acceptable salt thereof, wherein the constituent variables are defined herein.

[0009] The present disclosure also provides a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0010] The present disclosure also provides methods of treating a disease or condition (e.g., cancer) by administering a therapeutically effective amount of a compound of the present disclosure to a human or other mammal in need of such treatment. In some embodiments, the disease or condition is characterized by acidic or hypoxic diseased tissue.

[0011] The present disclosure also provides uses of the compounds described herein for the manufacture of a medicament for use in therapy. The present disclosure also provides compounds described herein for use in therapy.

[0012] The present disclosure also provides methods for synthesizing the disclosed compounds and intermediates useful in these methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Graph showing the effect of free DM4 and compound 5 on in vitro β-tubulin polymerization (in terms of relative fluorescence units) at different concentrations.

[0014] Figure 2 Depicted are kinetic analysis of compound 5 binding to β-tubulin in vitro as determined by Biacore surface plasmon resonance.

[0015] Figure 3A Graph showing mean tumor volume of nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5.

[0016] Figure 3B Shown are the percentage changes in body weight relative to day 0 in nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5.

[0017] Figure 4 Kaplan-Meier plots of nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5 are depicted.

[0018] Figure 5A Depicted are abdominal views and extracted lungs of nude mice inoculated with 4T1-RFP fluorescent cells via tail vein injection and imaged 11 days post-inoculation and after 3 doses of vehicle or compound 6.

[0019] Figure 5B Graph depicting fluorescent signals from extracted lungs of mice vaccinated with 4T1-RFP after 3 doses of vehicle or compound 6. DETAILED DESCRIPTION

[0020] Provided herein is a compound of formula (I):

[0021] R 2 -LR 1 (I)

[0022] or a pharmaceutically acceptable salt thereof, wherein:

[0023] R 1 It is a peptide;

[0024] R 2 is a small molecule microtubule targeting moiety; and

[0025] L is a linker that is covalently attached to the moiety R 1 and R 2 .

[0026] Provided herein is a compound of formula (I):

[0027] R 2 -LR 1 (I)

[0028] or a pharmaceutically acceptable salt thereof, wherein:

[0029] R 1 are peptides with 5 to 50 amino acids;

[0030] R 2 is a small molecule microtubule targeting moiety; and

[0031] L is a linker that is covalently attached to the moiety R 1 and R 2 .

[0032] Also provided herein is a compound of formula (I):

[0033] R 2 -LR 1 (I)

[0034] or a pharmaceutically acceptable salt thereof, wherein:

[0035] R 1 is able to selectively deliver R 2 L-peptides that cross the membranes of cells with acidic or anoxic mantles;

[0036] R 2 is a small molecule microtubule targeting moiety; and

[0037] L is a linker that is covalently attached to the moiety R 1 and R 2 .

[0038] In some embodiments, R 2 It is a maytansine-derived microtubule-targeting moiety.

[0039] Provided herein is a compound of formula (I):

[0040] R 2 -LR 1 (I)

[0041] or a pharmaceutically acceptable salt thereof, wherein:

[0042] R 1 is able to selectively deliver R 2 L-peptides that cross the membranes of cells with acidic or anoxic mantles;

[0043] R 2 Selected from the group consisting of:

[0044]

[0045]

[0046] and

[0047] L is a linker that is covalently attached to the moiety R 1 and R 2 .

[0048] Provided herein is a compound of formula (I):

[0049] R 2 -LR 1 (I)

[0050] or a pharmaceutically acceptable salt thereof, wherein:

[0051] R 1 is able to selectively deliver R 2 L-peptides that cross the membranes of cells with acidic or anoxic mantles;

[0052] R 2 Selected from the group consisting of:

[0053]

[0054] and

[0055] L is a linker that is covalently attached to the moiety R 1 and R 2 .

[0056] Provided herein is a compound of formula (I):

[0057] R2 -LR 1 (I)

[0058] or a pharmaceutically acceptable salt thereof, wherein:

[0059] R 1 is able to selectively deliver R 2 L-peptides that cross the membranes of cells with acidic or anoxic mantles;

[0060] R 2 Selected from the group consisting of:

[0061]

[0062]

[0063]

[0064] L is a group selected from the following:

[0065]

[0066]

[0067] in

[0068] R 3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 and R 10 Each independently selected from H, C 1-4 Alkyl, C 1-4 Alkenyl, C 6-10 Aryl, C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 4-10 membered heterocycloalkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NRc1 R d1 , wherein the C 1-4 Alkyl, C 1-4 Alkenyl, C 6-10 Aryl and 5-10 membered heteroaryl are each optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0069] or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0070] or R 3 and R 5 Together with the carbon atom to which it is attached, it forms C 3-14cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0071] or R 4 and R 6 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0072] or R 5 and R 6 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, ORa1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0073] or R 7 and R 8 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0074] or R 7 and R 9 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 Rd1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0075] or R 8 and R 10 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0076] or R 9 and R 10 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 Rd1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0077] Z is C 6-10 aryl or 5-10 membered heteroaryl; wherein the 5-10 membered heteroaryl has at least one ring-forming carbon atom and 1, 2, 3 or 4 ring-forming heteroatoms independently selected from N, O and S, wherein the C 6-10 Aryl and 5-10 membered heteroaryl are each optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0078] A is H or C 1-4 alkyl;

[0079] R a1 、R b1 、R c1 and R d1 Each independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, OH, CN, NO2 and CO2CH3; wherein the C 1-6 Alkyl and C 2-6 each alkenyl group is optionally substituted with OH, CN, NO2 or CO2CH; and

[0080] n is 0, 1, or 2.

[0081] Provided herein is a compound of formula (I):

[0082] R 2 -LR 1 (I)

[0083] or a pharmaceutically acceptable salt thereof, wherein:

[0084] R 1 is able to selectively deliver R 2 L-peptides that cross the membranes of cells with acidic or anoxic mantles;

[0085] R 2 Selected from the group consisting of:

[0086]

[0087]

[0088] L is a group selected from the following:

[0089]

[0090] in

[0091] R 3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 and R 10 Each independently selected from H, C 1-4 Alkyl, C 1-4 Alkenyl, C 6-10 Aryl, 5-10 membered heteroaryl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 , wherein the C 1-4 Alkyl, C1-4 Alkenyl, C 6-10 Aryl and 5-10 membered heteroaryl are each optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0092] or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0093] or R 3 and R 5 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NRc1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0094] or R 4 and R 6 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0095] or R 5 and R 6 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NRc1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0096] or R 7 and R 8 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0097] or R 7 and R 9 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0098] or R 8 and R 10 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0099] or R 9 and R 10 Together with the carbon atom to which it is attached, it forms C 3-7 Cycloalkyl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0100] A is H or C 1-4 alkyl; and

[0101] R a1 、R b1 、R c1 and R d1 Each independently selected from H, C 1-6 Alkyl, C2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, OH, CN, NO2 and CO2CH3; wherein the C 1-6 Alkyl and C 2-6 The alkenyl groups are each optionally substituted with OH, CN, NO2 or CO2CH.

[0102] In some embodiments, the left hand side of L is connected to R 2 and the right hand side of L is connected to R 1 .

[0103] In some embodiments, the sulfur atom of the disulfide portion of L is R 1 The portion of cysteine ​​residues.

[0104] As used herein, "peptide" refers to a targeting moiety comprising a 10-50 amino acid sequence consisting of naturally occurring amino acid residues and optionally one or more non-naturally occurring amino acids. 1 The peptides are peptides of 20 to 40, 20 to 30 amino acids, or 30 to 40 residues. Peptides suitable for use in the compounds of the present invention are those that can be inserted through the cell membrane via conformational changes or secondary structural changes in response to changes in environmental pH. In this way, the peptides can target acidic tissues and selectively translocate polar, cell-impermeable molecules across the cell membrane in response to low extracellular pH. In some embodiments, the peptides are capable of selectively translocating a conjugated moiety (e.g., R 2 L-) through cell membranes having an acidic or anoxic mantle with a pH less than about 6.0. In some embodiments, the peptide is capable of selectively transferring a conjugated moiety (e.g., R 2 L-) through cell membranes having an acidic or anoxic mantle with a pH less than about 6.5. In some embodiments, the peptide is capable of selectively transferring a conjugated moiety (e.g., R 2 L-) through cell membranes having an acidic or anoxic mantle with a pH less than about 5.5. In some embodiments, the peptide is capable of selectively transferring a conjugated moiety (e.g., R 2 L-) delivery across cell membranes having an acidic or anoxic mantle with a pH between about 5.0 and about 6.0.

[0105] In certain embodiments, R 1 The peptide includes a cysteine ​​residue that can form a bond with the payload moiety to be delivered across the cell membrane (e.g., R 2 In some embodiments, R 1 Via R 1The cysteine ​​residue of is linked to L. In some embodiments, the sulfur atom of the cysteine ​​residue can form part of a disulfide bond of the disulfide-containing linker L.

[0106] Suitable peptides that can conformationally change based on pH and insert across the cell membrane are described, for example, in U.S. Patents 8,076,451 and 9,289,508 (each of which is incorporated herein by reference in its entirety). Other suitable peptides are described, for example, in Weerakkody et al., PNAS 110(15), 5834-5839 (April 9, 2013), which is also incorporated herein by reference in its entirety.

[0107] In some embodiments, R 1 is a peptide comprising at least one of the following sequences:

[0108] ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO.1; Pv1),

[0109] AEQNPIYWARYADWLFFTTPLLLLDLALLVDADECG (SEQ ID NO. 2; Pv2), and

[0110] ADDQNPWRAYLDLLFPTDTLLLLDLLWDADECG(SEQ ID NO.3;Pv3);

[0111] Ac-AAEQNPIYWARYADWLFTTTPLLLLDLALLVDADEGTKCG (SEQ ID NO.4; Pv4);

[0112] AAEQNPIYWARYADWLFFTTPLLLLDLALLVDADEGTC (SEQ ID No. 5; Pv5); and

[0113] AAEQNPIYWWARYADWLFTTTPLLLLDLALLVDADEGTCG (SEQ ID No. 6; Pv6);

[0114] where R 1 Via R 1 The cysteine ​​residue of is connected to L.

[0115] In some embodiments, R 1 is a peptide comprising at least one of the following sequences:

[0116] ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO.1; Pv1),

[0117] AEQNPIYWARYADWLFFTTPLLLLDLALLVDADECG (SEQ ID NO. 2; Pv2), and

[0118] ADDQNPWRAYLDLLFPTDTLLLLDLLWDADECG (SEQ ID NO. 3; Pv3); and

[0119] AAEQNPIYWWARYADWLFTTTPLLLLDLALLVDADEGTCG (SEQ ID No. 6; Pv6);

[0120] where R 1 Via R 1 The cysteine ​​residue of is connected to L.

[0121] In some embodiments, R 1 It is a peptide comprising the sequence ADDQNPWRAYLDLLFPTD TLLLDLLWCG (SEQ ID NO. 1; Pv1).

[0122] In some embodiments, R 1 It is a peptide comprising the sequence AEQNPIYWARYADWLFTTP LLLLDLALLVDADECG (SEQ ID NO. 2; Pv2).

[0123] In some embodiments, R 1 It is a peptide comprising the sequence ADDQNPWRAYLDLLFPTD TLLLDLLWDADECG (SEQ ID NO. 3; Pv3).

[0124] In some embodiments, R 1 It is a peptide comprising the sequence Ac-AAEQNPIYWARYADWLFTTPLLLLDLALLVDADEGTKCG (SEQ ID NO. 4; Pv4).

[0125] In some embodiments, R 1 It is a peptide comprising the sequence AAEQNPIYWARYADWLFT TPLLLLDLALLVDADEGTC (SEQ ID NO. 5; Pv5).

[0126] In some embodiments, R 1 It is a peptide comprising the sequence AAEQNPIYWWARYADWLF TTPLLLLDLALLVDADEGTCG (SEQ ID NO. 6; Pv6).

[0127] In some embodiments, R 1It is a peptide consisting of the sequence ADDQNPWRAYLDLLFPTDTLLLDLLWCG (SEQ ID NO. 1; Pv1).

[0128] In some embodiments, R 1 It is a peptide consisting of the sequence AEQNPIYWARYADWLFTTPL LLLDLALLVDADECG (SEQ ID NO. 2; Pv2).

[0129] In some embodiments, R 1 It is a peptide consisting of the sequence ADDQNPWRAYLDLLFPTDTLLLDLLWDADECG (SEQ ID NO. 3; Pv3).

[0130] In some embodiments, R 1 It is a peptide consisting of the sequence Ac-AAEQNPIYWARYADWLFTTPLLLLDLALLVDADEGTKCG (SEQ ID NO. 4; Pv4).

[0131] In some embodiments, R 1 It is a peptide consisting of the sequence AAEQNPIYWARYADWLFTTP LLLLDLALLVDADEGTC (SEQ ID NO. 5; Pv5).

[0132] In some embodiments, R 1 It is a peptide consisting of the sequence AAEQNPIYWWARYADWLFT TPLLLLDLALLVDADEGTCG (SEQ ID NO. 6; Pv6).

[0133] In some embodiments, R 1 is a peptide comprising at least one sequence selected from SEQ ID NO: 7 to SEQ ID NO: 311 as shown in Table 1.

[0134] In some embodiments, R 1 is a peptide consisting of a sequence selected from SEQ ID NO: 7 to SEQ ID NO: 311 as shown in Table 1.

[0135] Table 1. Additional R 1 sequence

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142]

[0143]

[0144] Any of the described peptides useful in the present invention can be modified to include a cysteine ​​residue by replacing a non-cysteine ​​residue with a cysteine ​​residue or appending a cysteine ​​residue to the N-terminus or C-terminus.

[0145] In some embodiments, R 1 The peptides are conformationally constrained peptides. Conformationally constrained peptides may include, for example, macrocyclic peptides and stapled peptides. Stapled peptides are peptides constrained by covalent bonds between two amino acid side chains to form a peptide macrocycle. Conformationaly constrained peptides are described in, for example, Guerlavais et al., Annual Reports in Medicinal Chemistry 2014, 49, 331-345; Chang et al., Proceedings of the National Academy of Sciences of the United States of America (2013), 110(36), E3445-E3454; Tesauro et al., Molecules 2019, 24, 351-377; Dougherty et al., Journal of Medicinal Chemistry (2019), 62(22), 10098-10107; and Dougherty et al., Chemical Reviews (2019), 119(17), 10241-10287, each of which is incorporated herein by reference in its entirety.

[0146] In some embodiments, R 1 is a peptide having 10 to 50 amino acids. In some embodiments, R 1 is a peptide having 20 to 40 amino acids. In some embodiments, R 1 is a peptide having 20 to 40 amino acids. In some embodiments, R 1 is a peptide having 10 to 20 amino acids. 1 is a peptide having 20 to 30 amino acids. In some embodiments, R 1It is a peptide with 30 to 40 amino acids.

[0147] Suitable small molecule microtubule targeting moieties (e.g., R 2 ) may be a cytotoxic compound (such as a maytansine), which may have undesirable side effects when delivered systemically due to its possible adverse effects on normal tissues. Small molecule microtubule targeting agents include, but are not limited to, maytansines, aclitaxel, docetaxel, epothilones, discodermolide, vinca alkaloids, colchicine, combretastatins, and derivatives and analogs thereof. Microtubule targeting agents are described in Tangutur, AD, Current Topics in Medicinal Chemistry, 201717(22):2523-2537. Microtubule targeting agents also include maytansinoids (eg, maytansine (DM1) and its derivatives and analogs, which are described in Lopus, M, Cancer Lett., 2011, 307(2): 113-118; and Widdison, W., J. Med. Chem. 2006, 49: 4392-4408.

[0148] In some embodiments, R 2 The following groups:

[0149]

[0150] In some embodiments, R 2 The following groups:

[0151]

[0152] In some embodiments, R 2 The following groups:

[0153]

[0154] In some embodiments, R 2 The following groups:

[0155]

[0156] In some embodiments, R 2 The following groups:

[0157]

[0158] In some embodiments, R 2 In some embodiments, R 2 is DM1 or DM4. In some embodiments, R 2 is DM1. In some embodiments, R 2 It's DM4.

[0159] In some embodiments, L is covalently linked to R 1 and R 2 and is used to release R-containing proteins near acidic or hypoxic tissues (e.g., inside cells of diseased tissues). 2 part.

[0160] In some embodiments, L is a linking chain of 1 to 40, 1 to 30, 1 to 25, 1 to 20, 1 to 15, 1 to 10, or 1 to 5 chain atoms (including carbon and heteroatoms), optionally separated by 1-10 R q substituted with a substituent, and wherein one or more chain carbon atoms of L can be oxidized to form a carbonyl group (C=O), and wherein one or more chain atoms of N and S can each be optionally oxidized to form an amine oxide, a sulfoxide, or a sulfonyl group; wherein

[0161] Each R q Independently selected from OH, CN, -COOH, NH2, halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, C 1-6 Alkylthio, phenyl, 5-6 membered heteroaryl, 4-6 membered heterocycloalkyl, C 3-6 Cycloalkyl, NH(C 1-6 alkyl) and N(C 1-6 alkyl)2, wherein R q C 1-6 Alkyl, phenyl, C 3-6 Cycloalkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl are each optionally substituted with halo, OH, CN, -COOH, NH2, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy, phenyl, C 3-10 cycloalkyl, 5-membered or 6-membered heteroaryl, or 4-6-membered heterocycloalkyl; and

[0162] Two R's q The group together with the chain atoms to which it is attached can form a phenyl group, a 5-6 membered heteroaryl group, a 4-6 membered heterocycloalkyl group or a C 3-6 Cycloalkyl ring.

[0163] In some embodiments, R q Independently selected from OH, CN, -COOH, NH2, halogen, C 1-6 Haloalkyl, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, NH(C 1-6 alkyl) and N(C 1-6 Alkyl)2.

[0164] In some embodiments, L is the following group:

[0165]

[0166] In some embodiments, L is the following group:

[0167]

[0168] In some embodiments, L is the following group:

[0169]

[0170] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0171] In some embodiments, L is the following group:

[0172]

[0173] In some embodiments, L is the following group:

[0174]

[0175] In some embodiments, L is the following group:

[0176]

[0177] In some embodiments, L is the following group:

[0178]

[0179] In some embodiments, R 3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 and R 10 Each independently selected from H and C 1-4 In some embodiments, R3 、R 4 、R 5 、R 6 、R 7 、R 8 、R 9 and R 10 Each is H.

[0180] In some embodiments, R 3 and R 4 Each independently selected from H and C 1-4 In some embodiments, R 3 and R 4 Each is H.

[0181] In some embodiments, R 5 and R 6 Each independently selected from H and C 1-4 In some embodiments, R 5 and R 6 Each is H.

[0182] In some embodiments, R 7 and R 8 Each independently selected from H and C 1-4 In some embodiments, R 7 and R 8 Each is H.

[0183] In some embodiments, R 9 and R 10 Each independently selected from H and C 1-4 In some embodiments, R 9 and R 10 Each is H.

[0184] In some embodiments, A is H. In some embodiments, A is C 1-4 In some embodiments, A is CH3.

[0185] In some embodiments, Z is C 6-10 Aryl, optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 .

[0186] In some embodiments, Z is phenyl, optionally substituted with 1, 2, or 3 substituents independently selected from: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 .

[0187] In some embodiments, Z is phenyl.

[0188] In some embodiments, the compound of the present invention is a compound of formula (II):

[0189]

[0190] or a pharmaceutically acceptable salt thereof, wherein:

[0191] R 1 It is a peptide;

[0192] R 2 is the small molecule microtubule targeting moiety;

[0193] A is H or C 1-4 alkyl;

[0194] Ring Y is a monocyclic C 5-7 a cycloalkyl ring or a monocyclic 5-7 membered heterocycloalkyl ring;

[0195] Each R Y Independently selected from C 1-4 Alkyl, halogen, CN, NO2, ORa1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0196] or two adjacent R Y Together with the atoms to which it is attached, it forms a fused monocyclic ring C 5-7 Cycloalkyl ring, fused monocyclic 5-7 membered heterocycloalkyl ring, fused C 6-10 an aryl ring or a fused 6-10 membered heteroaryl ring, each of which is optionally substituted with 1, 2 or 3 substituents independently selected from: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ;

[0197] R a1 、R b1 、R c1 and R d1 Each independently selected from H, C 1-4 Alkyl, C 2-4 Alkenyl, C 2-4 alkynyl, each of which is optionally substituted with 1, 2 or 3 substituents independently selected from halo, OH, CN and NO2; and

[0198] m is 0, 1, 2 or 3.

[0199] In some embodiments of the compound of Formula (II), R 1 It is a peptide comprising the sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or SEQ ID NO: 5.

[0200] In some embodiments of the compound of Formula (II), R 1 It is Pv1, Pv2, Pv3, Pv4 or Pv5.

[0201] In some embodiments of the compound of Formula (II), R 1 Via R 1 A cysteine ​​residue of is linked to the core, wherein one of the sulfur atoms of the disulfide moiety in Formula II is derived from the cysteine ​​residue.

[0202] In some embodiments of the compound of Formula (II), R 2 In some embodiments of Formula (II), R 2 is DM1 or DM4. In some embodiments of formula (II), R 2 is DM1. In some embodiments of Formula (II), R 2 It's DM4.

[0203] In some embodiments of the compound of Formula (II), R 2 The following groups:

[0204]

[0205] In some embodiments of the compound of Formula (II), R 2 The following groups:

[0206]

[0207] In some embodiments of the compound of Formula (II), R 2 The following groups:

[0208] In some embodiments of the compound of Formula (II), R 2 The following groups:

[0209]

[0210] In some embodiments of the compound of formula (II), A is H. In some embodiments of the compound of formula (II), A is C 1-4In some embodiments of the compound of formula (II), A is CH3.

[0211] In some embodiments of the compound of Formula (II), Ring Y is a monocyclic C 5-7 Cycloalkyl ring.

[0212] In some embodiments of the compound of Formula (II), Ring Y is a cyclopentyl ring.

[0213] In some embodiments of the compound of Formula (II), Ring Y is a cyclohexyl ring.

[0214] In some embodiments of the compound of Formula (II), Ring Y is a cycloheptyl ring.

[0215] In some embodiments of the compound of Formula (II), Ring Y is a monocyclic 5-7 membered heterocycloalkyl ring.

[0216] In some embodiments of the compound of Formula (II), Ring Y is a 5-membered heterocycloalkyl ring.

[0217] In some embodiments of the compound of Formula (II), Ring Y is a 6-membered heterocycloalkyl ring.

[0218] In some embodiments of the compound of Formula (II), Ring Y is a 7-membered heterocycloalkyl ring.

[0219] In some embodiments of the compound of formula (II), two adjacent R Y Together with the atoms to which it is attached, it forms a fused monocyclic ring C 5-7 Cycloalkyl ring, fused monocyclic 5-7 membered heterocycloalkyl ring, fused C 6-10 an aryl ring or a fused 6-10 membered heteroaryl ring, each of which is optionally substituted with 1, 2 or 3 substituents independently selected from: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 .

[0220] In some embodiments of the compound of Formula (II), m is 0.

[0221] In some embodiments of the compound of Formula (II), m is 1.

[0222] In some embodiments of the compound of Formula (II), m is 2.

[0223] In some embodiments of the compound of Formula (II), m is 3.

[0224] In some embodiments, the compound of the present invention is a compound of Formula (III), Formula (IV), or Formula (V):

[0225]

[0226] or a pharmaceutically acceptable salt thereof, wherein R 1 、R 2 、R Y , A and m are as defined above in any of the embodiments of formula (II).

[0227] In some embodiments, the compound of formula (I) is selected from:

[0228]

[0229]

[0230]

[0231]

[0232]

[0233] or a pharmaceutically acceptable salt of any one of the foregoing.

[0234] In some embodiments, the compound of formula (I) is selected from:

[0235]

[0236]

[0237]

[0238] or a pharmaceutically acceptable salt of any one of the foregoing.

[0239] In some embodiments, provided herein is a compound having Formula (IA):

[0240]

[0241] or a salt thereof, wherein Cy 1 It is C 6-10 In some embodiments, Cy 1 It is pyridyl.

[0242] In some embodiments, provided herein is a compound having Formula (IB):

[0243]

[0244] or a salt thereof, wherein Cy 1 It is C 6-10 In some embodiments, Cy 1 It is pyridyl.

[0245] The molecules of the invention can be labeled, for example, with probes such as fluorophores, radioisotopes, etc. In some embodiments, the probe is a fluorescent probe, such as LICOR. A fluorescent probe can include any moiety (e.g., a fluorophore) that can re-emit light upon excitation.

[0246] The amino acids are represented by the following IUPAC abbreviations: alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine ​​(Cys; C), glutamine (Gln; Q), glutamic acid (Glu; E), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Val; V).

[0247] The term "Pv1" means ADDQNPWRAYLDLLFPTDTLLLDLLWCG, which is the peptide of SEQ ID No. 1.

[0248] The term "Pv2" means AEQNPIYWARYADWLFTTPLLLLDLALLVDAD ECG, which is the peptide of SEQ ID No. 2.

[0249] The term "Pv3" means ADDQNPWRAYLDLLFPTDTLLLDLLWDADEC G, which is the peptide of SEQ ID No. 3.

[0250] The term "Pv4" refers to Ac-AAEQNPIYWARYADWLFTTPLLLLDLALLV DADEGTKCG, which is the peptide of SEQ ID NO. 4.

[0251] The term "Pv5" means AAEQNPIYWARYADWLFTTPLLLLDLALLVDA DEGTC, which is a peptide of SEQ ID NO. 5. The term "Pv6" means AAEQNPIYWW ARYADWLFTTPLLLLDLALLVDADEGTCG, which is a peptide of SEQ ID NO. 6. In the compound of the present invention, the peptide R 1 It is linked to a disulfide linker via a cysteine ​​moiety.

[0252] The term "acidic and / or anoxic mantle" refers to the environment of the cells in the diseased tissue in question, which has a pH below 7.0 and preferably below 6.5. The acidic or anoxic mantle more preferably has a pH of about 5.5 and most preferably has a pH of about 5.0. The compounds of formula (I) insert in a pH-dependent manner through the cell membrane having an acidic and / or anoxic mantle to transfer R 2 L is inserted into the cell, whereupon the disulfide bond of the linker is cleaved to deliver the free R 2 L(or R 2 L*, wherein L* is a degradation product). Due to the pH dependence of the compounds of formula (I), they preferentially insert across cell membranes only in the presence of an acidic or anoxic mantle surrounding the cell and do not penetrate the cell membranes of "normal" cells that do not have an acidic or anoxic mantle.

[0253] As used herein, the term "pH sensitivity" or "pH dependence" refers to the 1 Or refers to peptide R 1 The mode of insertion of the compounds of the present invention through the cell membrane means that the peptide has a higher affinity for the cell membrane lipid bilayer with an acidic or anoxic mantle than for the membrane lipid bilayer at neutral pH. Therefore, when the cell membrane lipid bilayer has an acidic or anoxic mantle ("diseased" cells), the compounds of the present invention preferentially insert through the cell membrane to allow R 2 L is inserted into the cell interior (and thus delivers R as described above). 2 H), but does not penetrate the cell membrane when the mantle (the environment of the cell membrane lipid bilayer) is not acidic or hypoxic ("normal" cells). This preferential insertion is believed to be due to the fact that peptide R 1 This is achieved by forming a helical configuration, which facilitates membrane insertion.

[0254] The term "small molecule microtubule targeting moiety" refers to a chemical group that binds to microtubules. The small molecule microtubule targeting moiety can be a group derived from a compound that inhibits microtubule activity. For example, the small molecule microtubule targeting moiety can inhibit the dynamic stability of microtubules. In some embodiments, the small molecule microtubule targeting moiety has a molecular weight (Da) of about 100-1500, about 100-800, about 500-1,000, about 600-1,000, about 100-500, about 700-900, or about 250-500.

[0255] It will be further understood that certain features of the invention described in the context of separate embodiments for clarity may also be provided in combination in a single embodiment (with the embodiments intended to be combined as if written in multiple dependent forms). Conversely, various features of the invention described in the context of a single embodiment for simplicity may also be provided individually or in any suitable subcombination. Thus, it is contemplated that the features of the embodiments described as compounds of formula (I) may be combined in any suitable combination.

[0256] At various places in this specification, certain features of compounds are disclosed in groups or ranges. In particular, it is intended that this disclosure include each and every individual subcombination of the members of such groups and ranges. For example, in particular, the term "C 1-6 "Alkyl" is intended to individually disclose, but not be limited to, methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0257] The term "n-membered" (where n is an integer) generally describes the number of ring-forming atoms in a moiety, where the number of ring-forming atoms is n. For example, piperidinyl is an example of a 6-membered heterocycloalkyl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, pyridyl is an example of a 6-membered heteroaryl ring, and 1,2,3,4-tetrahydro-naphthalene is an example of a 10-membered cycloalkyl ring.

[0258] At various locations in this specification, variables defining a divalent linking group may be described. Specifically, it is intended that each linking substituent include both the forward and backward forms of the linking substituent. For example, -NR(CR'R") n -Include-NR(CR'R") n -and-(CR'R") n NR-, and it is intended that each of the forms be disclosed individually. Where a structure requires a linking group, the Markush variable listed for that group is to be understood as the linking group. For example, if a structure requires a linking group and the Markush group definition for that variable lists "alkyl" or "aryl," it is to be understood that "alkyl" or "aryl" refers to an alkylene linking group or an arylene linking group, respectively.

[0259] The term "substituted" means that an atom or a group of atoms formally replaces hydrogen as a "substituent" connected to another group. Unless otherwise indicated, the term "substituted" refers to any level of substitution, such as monosubstituted, disubstituted, trisubstituted, tetrasubstituted or pentasubstituted, where such substitutions are permitted. Substituents are independently selected, and substitutions can be located at any chemically accessible position. It should be understood that substitution at a given atom is limited by valence. It should be understood that substitution at a given atom produces a chemically stable molecule. The phrase "optionally substituted" means unsubstituted or substituted. The term "substituted" means that a hydrogen atom is removed and replaced by a substituent. A single divalent substituent (e.g., oxo) can replace two hydrogen atoms.

[0260] The term "C n-m " indicates a range including the endpoints, where n and m are integers and indicate the number of carbons. Examples include C 1-4 、C 1-6 wait.

[0261] The term "alkyl" used alone or in combination with other terms refers to a saturated hydrocarbon group which may be straight or branched. n-m "Alkyl" refers to an alkyl group having n to m carbon atoms. Alkyl formally corresponds to an alkane with one C-H bond replaced by the point of attachment of the alkyl group to the rest of the compound. In some embodiments, the alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl; higher carbon number homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like.

[0262] The term "alkenyl" used alone or in combination with other terms refers to a straight or branched hydrocarbon radical corresponding to an alkyl radical having one or more carbon-carbon double bonds. Alkenyl formally corresponds to an alkene in which one C—H bond is replaced by the point of attachment of the alkenyl group to the rest of the compound. n-m "Alkenyl" refers to an alkenyl group having n to m carbons. In some embodiments, the alkenyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, and the like.

[0263] The term "alkynyl", used alone or in combination with other terms, refers to a straight or branched hydrocarbon radical corresponding to an alkyl radical having one or more carbon-carbon triple bonds. Alkynyl formally corresponds to an alkyne in which one C—H bond is replaced by an alkyl radical at the point of attachment to the rest of the compound. The term "C n-m"Alkynyl" refers to an alkynyl group having n to m carbons. Example alkynyl groups include, but are not limited to, ethynyl, propyn-1-yl, propyn-2-yl, and the like. In some embodiments, the alkynyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms.

[0264] The term "alkylene," used alone or in combination with other terms, refers to a divalent alkyl linking group. Alkylene formally corresponds to an alkane with two C—H bonds replaced by alkylene groups at the point of attachment to the rest of the compound. n-m “Alkylene” refers to an alkylene radical having n to m carbon atoms. Examples of alkylene radicals include, but are not limited to, ethane-1,2-diyl, ethane-1,1-diyl, propylene-1,3-diyl, propylene-1,2-diyl, propylene-1,1-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, 2-methyl-propylene-1,3-diyl, and the like.

[0265] The term "amino" refers to a group of formula -NH2.

[0266] The term "carbonyl," employed alone or in combination with other terms, refers to a -C(=O)- group, which may also be written C(O).

[0267] The term "cyano" or "nitrile" refers to a group having the formula -C≡N, which may also be written as -CN.

[0268] The term "halo" or "halogen," used alone or in combination with other terms, refers to fluorine, chlorine, bromine, and iodine. In some embodiments, "halo" refers to a halogen atom selected from F, Cl, or Br. In some embodiments, halo is F.

[0269] As used herein, the term "haloalkyl" refers to an alkyl group in which one or more of the hydrogen atoms have been replaced by a halogen atom. n-m "Haloalkyl" refers to a C group having n to m carbon atoms and at least one and at most {2(nm)+1} halogen atoms. n-m In some embodiments, the halogen atom is a fluorine atom. In some embodiments, the haloalkyl group has 1 to 6 or 1 to 4 carbon atoms. Examples of haloalkyl groups include CF 3 , C 2 F 5 , CHF 2 , CH 2 F, CCl 3 , CHCl 2 , C 2 Cl 5 and the like. In some embodiments, the haloalkyl group is a fluoroalkyl group.

[0270] The term "haloalkoxy," used alone or in combination with other terms, refers to a radical of the formula -O-haloalkyl, wherein haloalkyl is as defined above. n-m"Haloalkoxy" refers to a haloalkoxy group wherein the haloalkyl group has n to m carbon atoms. Example haloalkoxy groups include trifluoromethoxy and the like. In some embodiments, the haloalkoxy group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0271] The term "oxo" refers to an oxygen atom as a divalent substituent which, when attached to carbon, forms a carbonyl group, or is attached to a heteroatom to form a sulfoxide or sulfone group, or an N-oxide group. In some embodiments, the heterocyclyl group may be optionally substituted with 1 or 2 oxo (=O) substituents.

[0272] The term "oxidized" with respect to a ring-forming N atom refers to a ring-forming N-oxide.

[0273] The term "oxidized" with respect to a ring-forming S atom refers to a ring-forming sulfonyl group or a ring-forming sulfinyl group.

[0274] The term "aromatic" refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings having aromatic character (ie, having (4n+2) delocalized π (pi) electrons, where n is an integer).

[0275] The term "aryl" used alone or in combination with other terms refers to an aromatic hydrocarbon group which may be monocyclic or polycyclic (eg, having two fused rings). n-m "Aryl" refers to an aromatic radical having n to m ring carbon atoms. Aryl groups include, for example, phenyl, naphthyl, and the like. In some embodiments, an aryl group has 6 to about 10 carbon atoms. In some embodiments, an aryl group has 6 carbon atoms. In some embodiments, an aryl group has 10 carbon atoms. In some embodiments, an aryl group is phenyl.

[0276] The term "heteroaryl" or "heteroaromatic" used alone or in combination with other terms refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from sulfur, oxygen, and nitrogen. In some embodiments, the heteroaryl ring has 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, any ring-forming N in the heteroaryl moiety may be an N-oxide. In some embodiments, the heteroaryl group has 5 to 14 ring atoms comprising carbon atoms and 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl group has 5 to 10 ring atoms comprising carbon atoms and 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl group has 5 to 6 ring atoms and 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl group is a five-membered or six-membered heteroaryl ring. In other embodiments, the heteroaryl group is an eight-, nine-, or ten-membered fused bicyclic heteroaryl ring.

[0277] A five-membered heteroaryl ring is a heteroaryl group having five ring atoms, wherein one or more (eg, 1, 2, or 3) of the ring atoms are independently selected from N, O, and S.

[0278] A six-membered heteroaryl ring is a heteroaryl group having six ring atoms, wherein one or more (eg, 1, 2, or 3) of the ring atoms are independently selected from N, O, and S.

[0279] The term "cycloalkyl," used alone or in combination with other terms, refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic) including cyclized alkyl and alkenyl groups. n-m "Cycloalkyl" refers to a cycloalkyl group having n to m ring member carbon atoms. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) groups and spirocyclic groups. Cycloalkyl groups may have 3, 4, 5, 6, or 7 ring carbon atoms (C 3-7 In some embodiments, the cycloalkyl group has 3 to 6 ring members, 3 to 5 ring members, or 3 to 4 ring members. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is monocyclic or bicyclic. In some embodiments, the cycloalkyl group is C 3-6 Monocyclic cycloalkyl. The ring-forming carbon atoms of cycloalkyl can be optionally oxidized to form oxo or sulfide groups. Cycloalkyl also includes cycloalkylene. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. The definition of cycloalkyl also includes parts with one or more aromatic rings fused to the cycloalkyl ring (i.e., having a common bond), such as benzo or thienyl derivatives of cyclopentane, cyclohexane, etc. The cycloalkyl containing the fused aromatic ring can be connected via any ring-forming atom of the ring-forming atoms including the fused aromatic ring. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, etc. In some embodiments, cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0280] The term "heterocycloalkyl" used alone or in combination with other terms refers to a non-aromatic ring or ring system, which may optionally contain one or more alkenylene groups as part of the ring structure, which has at least one heteroatom ring member independently selected from nitrogen, sulfur, oxygen and phosphorus, and which has 4-10 ring members, 4-7 ring members or 4-6 ring members. The term "heterocycloalkyl" includes monocyclic 4-, 5-, 6- and 7-membered heterocycloalkyls. Heterocycloalkyls may include monocyclic or bicyclic (e.g., having two fused or bridged rings) or spirocyclic ring systems. In some embodiments, heterocycloalkyls are monocyclic groups with 1, 2 or 3 heteroatoms independently selected from nitrogen, sulfur and oxygen. The ring-forming carbon atoms and heteroatoms of heterocycloalkyls may be optionally oxidized to form oxo or sulfide ion groups or other oxidized bonds (e.g., C(O), S(O), C(S) or S(O)2, N-oxides, etc.), or the nitrogen atom may be quaternized. Heterocycloalkyl can be connected via ring-forming carbon atoms or ring-forming heteroatoms. In some embodiments, heterocycloalkyl contains 0 to 3 double bonds. In some embodiments, heterocycloalkyl contains 0 to 2 double bonds. The definition of heterocycloalkyl also includes the part of the aromatic ring having one or more fused to the heterocycloalkyl ring (i.e., having a common bond), such as the benzo or thienyl derivatives of piperidine, morpholine, azacycloheptatriene (azepine) etc. The heterocycloalkyl containing fused aromatic ring can be connected via any ring-forming atom including the ring-forming atom of the fused aromatic ring. The example of heterocycloalkyl includes 2-pyrrolidinyl, morpholinyl, azetidinyl, tetrahydrofuranyl, tetrahydropyranyl and piperazinyl.

[0281] In certain positions, the definition or embodiment refers to a specific ring (e.g., an azetidine ring, a pyridine ring, etc.). Unless otherwise indicated, these rings may be connected to any ring member, provided that the valence of the atom is not exceeded. For example, the azetidine ring may be connected at any position of the ring, while the azetidine-3-yl ring is connected at the 3-position.

[0282] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are contemplated. Compounds of the present invention containing asymmetrically substituted carbon atoms can be separated in optically active or racemic form. Methods for preparing optically active forms from optically inactive starting materials are known in the art, for example, by resolving racemic mixtures or by stereoselective synthesis. Many geometric isomers of alkenes, C=N double bonds, etc. may also be present in the compounds described herein, and all such stable isomers are encompassed by the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and can be separated in the form of isomeric mixtures or by separating isomeric forms.

[0283] The splitting of the racemic mixture of compound can be carried out by any one of many methods known in the art.A kind of method includes using chiral splitting acid to carry out fractional recrystallization, and the chiral splitting acid is an optically active salified organic acid.The suitable splitting agent for fractional recrystallization method is such as optically active acid, such as tartaric acid D and L form, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid or various optically active camphorsulfonic acids (such as α-camphorsulfonic acid).Other splitting agents suitable for fractional crystallization method include the stereoisomerically pure form (for example, S and R form, or diastereoisomerically pure form) of α-methylbenzylamine, 2-phenylglycinol, norephedrine (norephedrine), ephedrine (ephedrine), N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane etc.

[0284] The resolution of the racemic mixture can also be carried out by eluting with a column packed with an optically active resolving agent (eg, dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by those skilled in the art.

[0285] In some embodiments, the compounds of the present invention have the (R)-configuration. In other embodiments, the compounds have the (S)-configuration. In compounds with more than one chiral center, unless otherwise indicated, each of the chiral centers in the compound can independently be (R) or (S).

[0286] The compounds of the present invention also include tautomeric forms. Tautomeric forms are produced by the transposition of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include proton transfer tautomers in isomeric protonation states with the same empirical formula and total charge. Example proton transfer tautomers include keto-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in a heterocyclic ring system, such as 1H-imidazole and 3H-imidazole, 1H-1,2,4-triazole, 2H-1,2,4-triazole and 4H-1,2,4-triazole, 1H-isoindole and 2H-isoindole, and 1H-pyrazole and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0287] The compounds of the present invention may also include all isotopes of atoms that appear in the intermediates or final compounds. Isotopes include those atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted by atomic isotopes of natural or non-natural abundance. In some embodiments, the compound includes at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced or substituted by deuterium. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 deuterium atoms. Synthetic methods for including isotopes in organic compounds are known in the art (Alan F. Thomas, Deuterium Labeling in Organic Chemistry (New York, NY, Appleton-Century-Crofts, 1971); Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, The Renaissance of H / D Exchange, Angew. Chem. Int. Ed. 2007, 7744-7765; James R. Hanson, The Organic Chemistry of Isotopic Labelling, Royal Society of Chemistry, 2011). Isotopically labeled compounds are useful in various studies, such as NMR spectroscopy, metabolic experiments and / or assays.

[0288] Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances (A. Kerekes et al., J. Med. Chem. 2011, 54, 201-210; R. Xu et al., J. Label Compd. Radiopharm. 2015, 58, 308-312).

[0289] As used herein, the term "compound" is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. The term also refers to a compound of the invention, regardless of how it is prepared, for example, synthetically, via a biological process (e.g., metabolic or enzymatic conversion), or a combination thereof.

[0290] All compounds and pharmaceutically acceptable salts thereof may exist together with other substances such as water and solvents (e.g., hydrates and solvates) or may be separated. When in the solid state, the compounds described herein and their salts may exist in various forms and may, for example, be in the form of solvates (including hydrates). The compounds may be in any solid state form, such as polymorphs or solvates, and therefore, unless otherwise expressly indicated, references to compounds and their salts in this specification should be understood to encompass any solid state form of the compounds.

[0291] In some embodiments, the compound of the invention or its salt is substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which it is formed or detected. Partial isolation can include, for example, a composition enriched in the compound of the invention. Substantially isolated can include a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of the compound of the invention or its salt.

[0292] The phrase "pharmaceutically acceptable" is used herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, 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.

[0293] As used herein, the expressions "ambient temperature" and "room temperature" are understood in the art and generally refer to a temperature about the temperature of the room in which the reaction is carried out, such as the reaction temperature, for example, a temperature of about 20°C to about 30°C.

[0294] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. The term "pharmaceutically acceptable salt" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues (e.g., amines); alkali metal or organic salts of acidic residues (e.g., carboxylic acids); and the like. Pharmaceutically acceptable salts of the present invention include, for example, non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., (Mack Publishing Company, Easton, 1985), p. 1418; Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19, and in Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In some embodiments, the compounds described herein include N-oxide forms.

[0295] synthesis

[0296] The compounds of the present invention, including salts thereof, can be prepared using known organic synthesis techniques and can be synthesized according to any of a number of possible synthetic routes, such as those in the schemes below.

[0297] The reaction for preparing the compounds of the present invention can be carried out in a suitable solvent that can be easily selected by a technician in the field of organic synthesis. At the temperature of the reaction (for example, the range can be the temperature of the solvent freezing temperature to the solvent boiling temperature), the suitable solvent can be substantially unreactive with the starting material (reactant), intermediate or product. A given reaction can be carried out in a solvent or a mixture of more than one solvent. Depending on the specific reaction step, the suitable solvent for the specific reaction step can be selected by a skilled person.

[0298] The preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups. One skilled in the art can readily determine the need for protection and deprotection and the selection of appropriate protecting groups. The chemistry of protecting groups is described, for example, in Kocienski, Protecting Groups, (Thieme, 2007); Robertson, Protecting Group Chemistry, (Oxford University Press, 2000); Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6th edition (Wiley, 2007); Peturssion et al., "Protecting Groups in Carbohydrate Chemistry", J. Chem. Educ., 1997, 74(11), 1297; and Wuts et al., Protective Groups in Organic Synthesis, 4th edition, (Wiley, 2006).

[0299] The reaction can be monitored by any suitable method known in the art. For example, product formation can be monitored by spectroscopic means (e.g., nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C), infrared spectroscopy, spectrophotometry (eg, UV-visible), mass spectrometry), or by chromatography (eg, high performance liquid chromatography (HPLC) or thin layer chromatography (TLC)).

[0300] The following schemes provide general guidance related to the preparation of compounds of the present invention. Those skilled in the art will understand that the preparations shown in the schemes can be modified or optimized using common knowledge of organic chemistry to prepare a variety of compounds of the present invention.

[0301] Compounds of formula (I) may be prepared, for example, using the methods as illustrated in the following schemes.

[0302] Scheme 1: Synthesis of direct conjugates

[0303]

[0304] Contains inherent thiols (R 2 Small molecule microtubule targeting moieties (e.g., DM1 or DM4) containing thiol groups (e.g., DM1-SH) can be activated by forming a pyridyl disulfide (wherein X is, e.g., H, a halide, etc.) that can be reacted with a thiol-containing R in a disulfide exchange reaction. 1 The peptide is replaced to obtain the desired conjugate, wherein -SS- is the linking moiety L.

[0305] Scheme 2: Synthesis of direct conjugates

[0306]

[0307] Alternatively, R with an inherent thiol 1 The peptide can be activated by forming a pyridyl disulfide (II) which can be reacted with a thiol-containing R in a disulfide exchange reaction. 2 The substitutions are made to give the desired direct conjugates, wherein L is -SS-.

[0308] Scheme 3: Synthesis of Thioethylamine Amide Conjugates

[0309]

[0310] A protected ethylamine containing a thiol group that has been activated as a pyridyl disulfide V can react with a thiol-containing R in a disulfide exchange reaction. 2 The disulfide VII can be deprotected to give VIII and further reacted with maleimidopropionic acid IX in an acidic coupling reaction to give amide X. Amide X can be reacted with a thiol-containing peptide in a Michael addition to give the desired conjugate.

[0311] Scheme 4: Synthesis of Thiobutyramide Conjugates

[0312]

[0313] Thiol-containing butyric acid that has been activated as pyridyl disulfide XI can react with thiol-containing R in a disulfide exchange reaction. 2 The disulfide acid XII can be reacted with ethylaminomaleimide XIII in an acidic coupling reaction to give amide XIV. Amide XIV can be reacted with a thiol-containing peptide in a Michael addition to give the desired conjugate.

[0314] Peptide R 1Can use first by Merrifield in JACS, the 85th volume, the solid phase synthesis method described in 2149-2154 pages (1963) to prepare, although also can adopt other methods known in the art.Merrifield technology is well-known and is the common method for preparing peptide.The useful technology for solid phase peptide synthesis is described in several books (for example Bodanszky's text " Principles of Peptide Synthesis ", Springer Verlag 1984).This synthetic method relates to the progressive addition of protected amino acid to the growing peptide chain that is covalently bound to solid resin particles.Through this program, reagent and by-product are removed by filtering, thus eliminating the necessity of purifying intermediate.The general concept of this method depends on making the first amino acid of chain be connected to solid polymer by covalent bond, then adds subsequent protected amino acid one at a time in a stepwise manner until desired sequence is assembled.Finally, protected peptide is removed from solid resin support and protecting group is cracked off.

[0315] The amino acid can be linked to any suitable polymer. The polymer must be insoluble in the solvent used, must have a stable physical form that permits ready filtration, and must contain a functional group to which the first amino acid can be tightly attached via a covalent bond. A variety of polymers are suitable for this purpose, such as cellulose, polyvinyl alcohol, polymethyl methacrylate, and polystyrene.

[0316] How to use

[0317] Provided herein is the use of a compound of formula (I) in treating a disease (e.g., cancer or neurodegenerative disease). Another aspect of the present invention is the use of a compound of formula (I) in treating a disease (e.g., cancer) involving acidic or hypoxic diseased tissue. Hypoxia and acidosis are physiological markers of many disease processes (including cancer). In cancer, hypoxia is a mechanism that causes the development of an acidic environment within a solid tumor. Therefore, hydrogen ions must be removed from the cell (e.g., by a proton pump) to maintain normal pH within the cell. Due to this output of hydrogen ions, when compared to normal cells, the pH gradient of cancer cells on the cell membrane lipid bilayer increases and the pH in the extracellular environment is lower. A method of improving the efficacy and therapeutic index of cytotoxic agents is to make full use of this physiological characteristic to provide selective delivery of compounds to hypoxic cells within healthy tissues.

[0318] In these treatment methods, a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof can be administered as a single agent or in combination with other forms of therapy (e.g., ionizing radiation or cytotoxic agents in the case of cancer). In combination therapy, as will be appreciated by those skilled in the art, a compound of formula (I) can be administered before, simultaneously with, or after other treatment modalities. Any treatment method (single agent or in combination with other forms of therapy) can be administered as a course of treatment involving multiple doses or over a period of time.

[0319] Examples of cancers that can be treated using the compounds of the present disclosure include, but are not limited to, colorectal cancer, gastric cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, malignant melanoma of the skin or inside the eye, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, carcinoma of the endometrium, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, Disease), non-Hodgkin's lymphoma, esophageal cancer, small intestinal cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemia (including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia), childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or urethra cancer, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers (including those induced by asbestos), and combinations of said cancers.

[0320] In some embodiments, cancers that can be treated with the compounds of the present disclosure include bladder cancer, bone cancer, glioma, breast cancer (e.g., triple-negative breast cancer), cervical cancer, colon cancer, colorectal cancer, endometrial cancer, epithelial cancer, esophageal cancer, Ewing's sarcoma, pancreatic cancer, gallbladder cancer, stomach cancer, gastrointestinal tumors, head and neck cancer (upper aerodigestive cancer), intestinal cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer, adenocarcinoma), melanoma, prostate cancer, rectal cancer, clear cell renal carcinoma, skin cancer, stomach cancer, testicular cancer, thyroid cancer, and uterine cancer.

[0321] In some embodiments, cancers that can be treated with the compounds of the present disclosure include melanoma (e.g., metastatic malignant melanoma), renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory prostate adenocarcinoma), breast cancer, triple-negative breast cancer, colon cancer, and lung cancer (e.g., non-small cell lung cancer and small cell lung cancer). Additionally, the present disclosure includes refractory or recurrent malignancies whose growth can be inhibited using the compounds of the present disclosure.

[0322] In some embodiments, cancers that can be treated using the compounds of the present disclosure include, but are not limited to, solid tumors (e.g., prostate cancer, colon cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), hematological cancers (e.g., lymphoma, leukemia (e.g., acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML)), DLBCL, mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, or multiple myeloma), and combinations of such cancers.

[0323] In certain embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof can be used in combination with a chemotherapeutic agent, targeted cancer therapy, immunotherapy, or radiotherapy. The agent can be combined with the compound of the present invention in the form of a single dosage form, or the agent can be administered simultaneously or sequentially in the form of a separate dosage form. In some embodiments, when administered with a compound of formula (I) or a pharmaceutically acceptable salt thereof, the combination of the compound of formula (I) and the corresponding microtubule targeting agent (e.g., R 2 -H) The chemotherapeutic agent, targeted cancer therapy, immunotherapy or radiation therapy is less toxic to the patient than when administered in combination, for example by exhibiting reduced myelotoxicity.

[0324] Suitable chemotherapeutic or other anticancer agents include, for example, alkylating agents (including but not limited to nitrogen mustards, ethyleneimine derivatives, alkyl sulfonates, nitrosoureas, and triazenes), such as uracil mustard, chlormethine, cyclophosphamide, TM ), ifosfamide, melphalan, chlorambucil, pipobroman, triethylene-melamine, triethylenethiophosphamide, busulfan, carmustine, lomustine, streptozocin, dacarbazine, and temozolomide.

[0325] Other suitable agents for use in combination with the compounds of the present invention include: dacarbazine (DTIC), optionally with other chemotherapy drugs (e.g., carmustine (BCNU) and cisplatin); the "Dartmouth regimen," which consists of DTIC, BCNU, cisplatin, and tamoxifen; a combination of cisplatin, vinblastine, and DTIC; or temozolomide. The compounds according to the present invention may also be combined with immunotherapy drugs, including cytokines such as interferon alpha, interleukin 2, and tumor necrosis factor (TNF).

[0326] Suitable chemotherapeutic or other anti-cancer agents include, for example, antimetabolites (including but not limited to, folate antagonists, pyrimidine analogs, purine analogs, and adenosine deaminase inhibitors), such as methotrexate, 5-fluorouracil, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatine, and gemcitabine.

[0327] Suitable chemotherapeutic or other anticancer agents also include, for example, certain natural products and their derivatives (e.g., vinca alkaloids, antitumor antibiotics, enzymes, lymphokines, and epipodophyllotoxins), such as vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, ara-C, paclitaxel, and dapoxetine. TM ), mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, interferon (especially IFN-a), etoposide, and teniposide.

[0328] Other cytotoxic agents that can be administered in combination with the compounds of the invention include, for example, navelbene, CPT-11, anastrazole, letrazole, capecitabine, reloxafine, cyclophosphamide, ifosfamide, and droloxafine.

[0329] Also suitable are cytotoxic agents, such as epidophyllotoxin; antineoplastic enzymes; topoisomerase inhibitors; procarbazine; mitoxantrone; platinum coordination complexes, such as cisplatin and carboplatin; biological response modifiers; growth inhibitory agents; antihormonal therapeutics; leucovorin; tegafur; and hematopoietic growth factors.

[0330] Other anticancer agents include antibody therapeutics, such as trastuzumab (Herceptin); antibodies to costimulatory molecules (such as CTLA-4, 4-1BB, and PD-1); or antibodies to cytokines (IL-10, TGF-α, etc.).

[0331] Other anticancer agents also include those that block the migration of immune cells, such as antagonists to chemokine receptors, including CCR2 and CCR4.

[0332] Other anticancer agents also include those that enhance the immune system, such as adjuvants or adoptive T cell transfer.

[0333] Anti-cancer vaccines that can be administered in combination with the compounds of the present invention include, for example, dendritic cells, synthetic peptides, DNA vaccines, and recombinant viruses.

[0334] Other suitable agents for use in combination with the compounds of the invention include chemotherapy combinations such as platinum-based doublets (cisplatin or carboplatin plus gemcitabine; cisplatin or carboplatin plus docetaxel; cisplatin or carboplatin plus paclitaxel; cisplatin or carboplatin plus pemetrexed) or gemcitabine plus paclitaxel in combination with particles for use in lung cancer and other solid tumors.

[0335] The compounds of the present invention can be effectively combined with antihormonal agents for the treatment of breast cancer and other tumors. Suitable examples are antiestrogens, including but not limited to tamoxifen and toremifene; aromatase inhibitors, including but not limited to letrozole, anastrozole, and exemestane; adrenocortical steroids (e.g., prednisone); progestogens (e.g., megastrol acetate); and estrogen receptor antagonists (e.g., fulvestrant). Suitable antihormonal agents for the treatment of prostate cancer and other cancers can also be combined with the compounds of the present invention. These include antiandrogens, including but not limited to flutamide, bicalutamide, and nilutamide; luteinizing hormone-releasing hormone (LHRH) analogs, including leuprolide, goserelin, triptorelin, and histrelin; LHRH antagonists (e.g., degarelix); androgen receptor blockers (e.g., enzalutamide); and agents that inhibit androgen production (e.g., abiraterone).

[0336] The compounds of the present invention can be combined with or administered sequentially with other agents targeting membrane receptor kinases, particularly for patients who have developed primary or acquired resistance to targeted therapies. These therapeutic agents include inhibitors or antibodies against EGFR, Her2, VEGFR, c-Met, Ret, IGFR1, or Flt-3, and against cancer-associated fusion protein kinases such as Bcr-Abl and EML4-Alk. EGFR inhibitors include gefitinib and erlotinib, and EGFR / Her2 inhibitors include, but are not limited to, dacomitinib, afatinib, lapitinib, and neratinib. EGFR antibodies include, but are not limited to, cetuximab, panitumumab, and necitumumab. c-Met inhibitors can be used in combination with the compounds of the present invention. These include onartumzumab, tivantnib, and INC-280. Agents targeting Abl (or Bcr-Abl) include imatinib, dasatinib, nilotinib, and ponatinib, and those targeting Alk (or EML4-ALK) include crizotinib.

[0337] Angiogenesis inhibitors may be effective in combination with the compounds of the present invention in some tumors. These inhibitors include antibodies to VEGF or VEGFR or kinase inhibitors of VEGFR. Antibodies or other therapeutic proteins directed against VEGF include bevacizumab and aflibercept. Inhibitors of VEGFR kinases and other anti-angiogenesis inhibitors include (but are not limited to) sunitinib, sorafenib, axitinib, cediranib, pazopanib, regorafenib, brivanib, and vandetanib.

[0338] Activation of intracellular signaling pathways is common in cancer, and agents targeting components of these pathways have been combined with receptor-targeted agents to enhance efficacy and reduce drug resistance. Examples of agents that can be combined with the compounds of the present invention include inhibitors of the PI3K-AKT-mTOR pathway, inhibitors of the Raf-MAPK pathway, inhibitors of the JAK-STAT pathway, and inhibitors of protein chaperones and cell cycle progression.

[0339] Agents targeting PI3 kinase include, but are not limited to, pilaralisib, idelalisib, and buparlisib. mTOR inhibitors (e.g., rapamycin, sirolimus, temsirolimus, and everolimus) can be combined with the compounds of the present invention. Other suitable examples include, but are not limited to, vemurafenib and dabrafenib (Raf inhibitors) and trametinib, selumetinib, and GDC-0973 (MEK inhibitors). Inhibitors of one or more JAKs (e.g., ruxolitinib, baricitinib, tofacitinib), Hsp90 (e.g., tanespiramycin), cyclin-dependent kinases (e.g., palbociclib), HDACs (e.g., panobinostat), PARP (e.g., olaparib), and proteasome (e.g., bortezomib, carfilzomib) may also be combined with the compounds of the invention. Another example of a PARP inhibitor that may be combined with the compounds of the invention is talazoparib.

[0340] Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in standard literature. For example, the administration of many chemotherapeutic agents is described in "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference as if fully set forth.

[0341] The phrase "therapeutically effective amount" of a compound (therapeutic agent, active ingredient, drug, etc.) refers to the amount of the compound that is administered to a subject in need of therapy or treatment that alleviates symptoms, ameliorates symptoms, or slows the onset of a disease condition, according to clinically accepted standards for the condition or disorder being treated. For example, a therapeutically effective amount can be an amount that has been demonstrated to have the desired therapeutic effect in in vitro assays, in vivo animal assays, or clinical trials. A therapeutically effective amount can vary based on the specific dosage form, method of administration, treatment regimen, the specific disease or disorder being treated, the benefit / risk ratio, and the like, as well as numerous other factors.

[0342] The therapeutically effective amount can be obtained from clinical trials, animal models, or in vitro cell culture assays. It is known in the art that an effective amount suitable for human use can be calculated from an effective amount determined from an animal model or in vitro cell culture assay. For example, as reported by Reagan-Shaw et al., FASEB J. 2008: 22(3)659-61, "μg / ml" (effective amount based on in vitro cell culture assay) = "mg / kg body weight / day" (effective amount for mice). In addition, based on the fact that the metabolic rate of mice is 6 times faster than that of humans, the effective amount for humans can be calculated from the effective amount for mice.

[0343] As an example of treatment using a combination of a compound of formula (I) and a cytotoxic agent, a therapeutically effective amount of a compound of formula (I) can be administered to a patient suffering from cancer as part of a treatment regimen that also involves a therapeutically effective amount of ionizing radiation or a cytotoxic agent. In the context of such a treatment regimen, the term "therapeutically effective" amount should be understood to mean an amount that is effective in combination therapy. Those skilled in the art of cancer treatment will understand how to adjust the dosage to achieve optimal therapeutic results.

[0344] Similarly, one skilled in the medical arts can readily determine appropriate dosages of the compounds of the present invention for the treatment of non-cancerous diseases or disorders, such as cardiovascular disease.

[0345] As used herein, the term "treatment" includes administering a compound or composition that reduces the frequency of symptoms of a disease (e.g., cancer, stroke, myocardial infarction, or long-term neurodegenerative disease) involving acidic or hypoxic diseased tissue in a subject relative to a subject who has not received the compound or composition, postpones its onset, or reduces its progression. This may include reversing, reducing, or inhibiting the underlying pathology of symptoms, clinical signs, or illness in a manner that improves or stabilizes the subject's illness (e.g., tumor growth, regression of cancer, or reduction or improvement of myocardial ischemia-reperfusion injury in myocardial infarction, stroke, or similar cardiovascular disease). The term "inhibit" or "reduce" is used for cancer and refers to a method of inhibiting or reducing tumor growth (e.g., reducing tumor size) in a colony compared to an untreated control colony.

[0346] All publications (including patents) mentioned herein are incorporated herein by reference for the purpose of describing and disclosing, for example, the constructs and methods described in the publications, which may be used in conjunction with the disclosure described herein. The publications discussed throughout are provided solely for their disclosure prior to the filing date of the present application.

[0347] Several types of ranges are disclosed herein. When any type of range is disclosed or claimed, it is intended to disclose or claim each possible number that this range can reasonably encompass, including the endpoints of the range and any subranges and combinations of subranges encompassed therein. For example, when the range of a therapeutically effective amount of an active ingredient is disclosed or claimed, it is intended to disclose or claim each possible number that the range can encompass, consistent with the disclosure herein. For example, the therapeutically effective amount of a disclosed compound can be in the range of about 1 mg / kg to about 50 mg / kg (subject body weight).

[0348] Formulation, dosage form and administration

[0349] To prepare the pharmaceutical composition of the present invention, the compound of formula (I) or a pharmaceutically acceptable salt thereof is used as the active ingredient in a uniform mixture according to conventional pharmaceutical compounding techniques, and is combined with a pharmaceutical carrier, which can be in a wide variety of forms depending on the form of preparation required for administration (e.g., oral or parenteral). When preparing a composition in an oral dosage form, any of the commonly used pharmaceutical media can be used, such as water, glycols, oils, alcohols, flavoring agents, preservatives, colorants, etc. in the case of oral liquid preparations such as suspensions, elixirs, and solutions; or carriers such as starch, sugar, diluents, granulating agents, lubricants, binders, disintegrants, etc. in the case of oral solid preparations such as powders, capsules, and tablets. Since tablets and capsules are easy to administer, they represent the most advantageous oral unit dosage form, in which case solid pharmaceutical carriers are obviously used. If necessary, tablets can be coated with sugar or enteric coating by standard techniques. For parenteral administration, the carrier will generally contain sterile water, but may include other ingredients, such as for promoting dissolution or preservative purposes. Injectable suspensions may also be prepared, in which case appropriate liquid carriers, suspending agents, etc. may be employed. Those skilled in the pharmaceutical and medical arts will readily be able to determine appropriate dosages of the pharmaceutical compositions of the present invention for the particular disease or condition to be treated.

[0350] Example

[0351] As used herein, all abbreviations, symbols, and conventions are consistent with those used in contemporary scientific literature. See, for example, Janet S. Dodd, ed., The ACS Style Guide: A Manual for Authors and Editors, 2nd ed., Washington, DC: American Chemical Society, 1997. The following definitions describe the terms and abbreviations used herein:

[0352] ●Brine: water saturated with NaCl solution

[0353] DCM: dichloromethane

[0354] TFA: trifluoroacetic acid

[0355] ●DIPEA: diisopropylethylamine

[0356] DMA: dimethylacetamide

[0357] ●DME: dimethoxyethane

[0358] DMF: dimethylformamide

[0359] DMSO: methyl sulfoxide

[0360] DTT: dithiothreitol

[0361] MSD: Mass spectrometry detector

[0362] Et2O: ethyl ether

[0363] EtOAc: ethyl acetate

[0364] EtOH: ethanol

[0365] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate

[0366] HOBt: 1-hydroxybenzotriazole

[0367] RP: Reverse Phase

[0368] HPLC: High-performance liquid chromatography

[0369] IPA: Isopropyl alcohol

[0370] LAH: lithium aluminum hydride

[0371] ●N-BuLi: n-butyllithium

[0372] LC-MS: Liquid chromatography-mass spectrometry

[0373] LDA: lithium diisopropylacetamide

[0374] Me: methyl

[0375] MeOH: methanol

[0376] MTBE: methyl tert-butyl ether

[0377] NMP: N-methylpyrrolidine

[0378] Ph: phenyl

[0379] ●PNPC: p-Nitrophenyl chloroformate

[0380] RT or rt: room temperature

[0381] SFC: Supercritical fluid chromatography

[0382] TBAI: Tetrabutylammonium iodide

[0383] TBME: tert-butyl methyl ether

[0384] tBu: tert-butyl

[0385] THF: tetrahydrofuran

[0386] TEA: triethylamine

[0387] TMEDA: Tetramethylethylenediamine

[0388] GSH: glutathione

[0389] GS: Sulfur-bound glutathione

[0390] LiOH: lithium hydroxide

[0391] DPPA: diphenylphosphoryl azide

[0392] Sn(Bu)2(laurate)2: dibutyltin dilaurate

[0393] PBS: Phosphate-buffered saline

[0394] ACN: acetonitrile

[0395] AcOH: acetic acid

[0396] ●EEDQ: N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline

[0397] DMAP: 4-dimethylaminopyridine

[0398] ●EDC: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0399] The HPLC method employed is described below.

[0400] HPLC method

[0401] A: Sunfire C18 150×4.6 mm; H 2 O / acetonitrile containing TFA modifier (0.05%); flow rate: 1 ml / min; wavelength = 217 nM.

[0402] B: Ace Equivalence 250×4.6 mm; H 2 O / acetonitrile containing TFA modifier (0.05%); flow rate: 1 ml / min; wavelength = 217 nM.

[0403] C: Sunfire C18 150×30 mm; H 2 O / acetonitrile containing TFA modifier (0.05%); flow rate: 30 ml / min; wavelength = 217 nM.

[0404] D: Sunfire C18 150×4.6 mm; H2O / acetonitrile containing AcOH modifier (0.5%); flow rate: 1 ml / min; wavelength = 217 nM

[0405] E: Sunfire C18 150×30 mm; H 2 O / acetonitrile containing AcOH modifier (0.5%); flow rate: 30 ml / min; wavelength = 217 nM.

[0406] F: Agilent 1100 / 1200 / 1260 or 1290 system (with or without MS).

[0407]

[0408]

[0409] G: Agilent 1100 / 1200 / 1260 or 1290 system (with or without MS).

[0410]

[0411] Mass spectrometry methods

[0412] Maldi-TOF (matrix-assisted laser desorption / ionization-time of flight) mass spectra were measured on an Applied Biosystems Voyager System 6268. Samples were prepared as a matrix of α-cyanohydroxycinnamic acid on AB Science plates (part number V700666).

[0413] Electrospray ionization (ESI) mass spectra were measured on an Agilent 1100 series LC-MS with a 1946 MSD or a Waters Xevo Qtof high resolution MS, both providing mass / charge species (m / z = 3).

[0414] The sources of the starting materials used in the examples are described in the table below.

[0415] Table 2. R 2 Starting materials

[0416]

[0417]

[0418] Intermediate I(R 2 Synthesis of SS-Pyr

[0419]

[0420] To (2S)-2-[methyl(3-thiopropionyl)amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (46.7 mg, 0.06 mmol) in 1 mL of CH3CN was added 2-(2-pyridyldisulfanyl)pyridine (20.0 mg, 0.09 mmol). The mixture was concentrated and purified (SiO2, 0-10% MeOH / CH2Cl2) to afford (2S)-2-[methyl-[3-(2-pyridyldisulfanyl)propanoyl]amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (53.6 mg, 100% yield). MS m / z 847.1 [M+H] + .

[0421] Synthesis of Pv3-S-Pyr (Intermediate II-3)

[0422]

[0423] Pv3 (250 mg, 0.06 mmol; in the form of a free-flowing solid) and 2-(2-pyridyldisulfanyl)pyridine (0.110 g, 0.5 mol) were dissolved in MeOH (10 mL) and the reaction was stirred at room temperature overnight. LC-MS indicated the formation of the desired product. The reaction mixture was concentrated and the residue was dissolved in DMSO and purified by reverse phase column chromatography (40-65% CH 3 CN / H 2 O (0.5% AcOH), 13 min) to give 212 mg of the desired product (187 mg, yield: 74.9%). MS m / z = 3 1273.4.

[0424] Intermediates II-1, II-2 and II-6 were prepared similarly to II-3 using Pv1, Pv2 and Pv6 as shown below:

[0425]

[0426] Table 3. Starting materials for L groups

[0427]

[0428]

[0429] Synthesis of intermediate VI-2

[0430]

[0431] 1-Amino-2-methyl-propane-2-thiol hydrochloride (100mg, 0.706mmol) is dissolved in CH2Cl2 (7mL) and 9H-fluorene-9-ylmethyl chloroformate (274mg, 1.06mmol) and N, N-diisopropylethylamine (182mg, 1.41mmol) are added thereto. The reaction mixture is stirred at room temperature overnight. The reaction mixture is washed with water and concentrated. The residue is purified by column chromatography (0-50% EtOAc / hexane) to obtain 9H-fluorene-9-ylmethyl N-(2-methyl-2-sulfanyl-propyl)carbamate (213mg, yield: 92.2%). MS m / z 350.1[M+Na] + .

[0432] Synthesis of intermediate V-1

[0433]

[0434] 2-(2-pyridyldisulfanyl)pyridine (746 mg, 3.38 mmol) was dissolved in MeOH (15 mL) and tert-butyl N-(2-thioethyl)carbamate (200 mg, 1.13 mmol) was added thereto. The reactant was stirred at room temperature for 3 h. The mixture was concentrated and the residue was purified by column chromatography (0-50% EtOAc / hexane) to give tert-butyl N-[2-(2-pyridyldisulfanyl)ethyl]carbamate (200 mg, yield: 61.9%). MS m / z 287.1[M+H] + .

[0435] Synthesis of intermediate V-2

[0436]

[0437] 2-(2-pyridyldisulfanyl)ethylamine hydrochloride (200 mg, 0.898 mmol) is dissolved in CH2Cl2 and 9H-fluoren-9-ylmethyl chloroformate (348 mg, 1.35 mmol) and N, N-diisopropylethylamine (232 mg, 1.80 mmol) are added thereto. The reaction mixture is stirred for 2 h at RT, washed with water and concentrated. The residue is purified by column chromatography (0-50% EtOAc / hexane) to obtain 9H-fluoren-9-ylmethyl N-[2-(2-pyridyldisulfanyl)ethyl]carbamate (288 mg, yield: 78.5%). MS m / z 409.1[M+H] + .

[0438] Synthesis of intermediate VII-1

[0439]

[0440] To a 1 mL flask containing (2S)-2-[methyl(3-thiopropionyl)amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (25.0 mg, 0.03 mmol) was added. To a vial of CH3CN was added tert-butyl N-[2-(2-pyridyldisulfanyl)ethyl]carbamate (Intermediate V-1, 14.5 mg, 0.051 mmol) and 4-methylmorpholine (0.138 mL, 1.25 mmol). The mixture was stirred for 16 h. LC-MS analysis indicated the desired material. The mixture was concentrated, dissolved in 50 mL of EtOAc and washed with 1 × 25 mL of saturated NH4Cl and 1 × 25 mL of saturated brine. The organic phase was dried over MgSO4, filtered and concentrated. The crude residue was purified (SiO2, 0-100% EtOAc / hexanes) to afford (2S)-2-[3-[2-(tert-butoxycarbonylamino)ethyldisulfanyl]propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (30.9 mg, 100% yield). MS m / z 913.2 [M+H] + .

[0441] Synthesis of intermediate VII-2

[0442]

[0443] Intermediate VII-2 was prepared similarly to VII-1 using Intermediate V-2 instead of Intermediate V-1.

[0444] Synthesis of intermediate VII-3

[0445]

[0446] To a 1 mL flask containing (2S)-2-[3-[2-(9H-fluoren-9-ylmethoxycarbonylamino)ethyldisulfanyl]propanoyl-methyl-amino]propionic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (25.0 mg, 0.03 mmol) was added. To a vial of CH3CN was added 9H-fluoren-9-ylmethyl N-(2-methyl-2-sulfanyl-propyl)carbamate (14.5 mg, 0.044 mmol) and 4-methylmorpholine (0.120 mL, 1.09 mmol). The mixture was stirred for 16 h. LC-MS analysis indicated the formation of the desired material. The mixture was concentrated, dissolved in 50 mL of EtOAc and washed with 1 × 25 mL of saturated NH4Cl and 1 × 25 mL of saturated brine. The organic phase was dried over MgSO4, filtered and concentrated. The crude residue was purified (SiO2, 0-100% EtOAc / hexanes) to (2S)-2-[3-[[2-(9H-fluoren-9-ylmethoxycarbonylamino)-1,1-dimethyl-ethyl]disulfanyl]propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (0.0313 g, 100% yield). MS m / z 1085.0[M+Na] + .

[0447] Synthesis of intermediate VIII-1 (BOC deprotection)

[0448]

[0449] (2S)-2-[3-[2-(tert-Butoxycarbonylamino)ethyldisulfanyl]propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (31.9 mg, 0.05 mmol) was dissolved in 0.3 / 0.1 / 0.1 mL CH3CN / H2O / TFA. The mixture was stirred for 36 h. LC-MS indicated complete deprotection. The mixture was purified by preparative HPLC (20-95% CH3CN / H2O containing 0.05% TFA) to give (2S)-2-[3-(2-aminoethyldisulfanyl)propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester]; 2,2,2-trifluoroacetic acid (22.9 mg, yield: 70.7%). MS m / z 813.2 [M+H] + .

[0450] Alternative synthesis of intermediate VIII-1 (FMOC deprotection)

[0451]

[0452] To a vial containing (2S)-2-[3-[2-(9H-fluoren-9-ylmethoxycarbonylamino)ethyldisulfanyl]propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (Intermediate VII-2; 29.6 mg, 0.03 mmol) was added 0.5 mL of DMF and 4-methylmorpholine (0.120 mL, 1.09 mmol). The mixture was heated at 40°C for 16 h. LC-MS confirmed complete deprotection. The mixture was purified by (20-95% CH3CN / H2O containing 0.05% TFA) to give (2S)-2-[3-(2-aminoethyldisulfanyl)propanoyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] trifluoroacetate (22.9 mg, yield: 86.4%). MS m / z 813.2[M+H] + .

[0453] Synthesis of intermediate VIII-2

[0454]

[0455] Intermediate VIII-2 was prepared similarly to Intermediate VIII-1. MS m / z 841.2 [M+H] + .

[0456] Synthesis of intermediate X-1

[0457]

[0458] To a 1 mL flask containing (2S)-2-[3-(2-aminoethyldisulfanyl)propionyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] trifluoroacetate (Intermediate VIII-1; 45.8 mg, 0.05 mmol) was added 1 mL flask containing (2S)-2-[3-(2-aminoethyldisulfanyl)propionyl-methyl-amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] trifluoroacetate (Intermediate VIII-1; 45.8 mg, 0.05 mmol) 3-(2,5-Dioxopyrrol-1-yl)propanoic acid (12.5 mg, 0.074 mmol), TBTU (23.8 g, 0.074 mmol), and DIPEA (0.0169 mL, 0.1 mmol) were added to DMF. LC-MS indicated complete conversion to product. The mixture was diluted with 50 mL of EtOAc. The mixture was washed with 1 x 25 mL of saturated NH4Cl, 4 x 25 mL of H2O, and 1 x 25 mL of H2O. The organic phase was dried over MgSO4, filtered, and concentrated. The crude product was purified (SiO2, 0-10% MeOH / Cl2Cl2) to give (2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propionylamino]ethyldisulfanyl]propionyl-methylamino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (17.3 mg, 36.5% yield) MS m / z 986.1 [M+Na] + .

[0459] Example 2: Synthesis of Compound 2

[0460]

[0461] To a vial containing Pv2 (25.0 mg, 0.006 mmol; as a free-flowing solid) and (2S)-2-[methyl-[3-(2-pyridyldisulfanyl)propanoyl]amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (7.70 mg, 0.009 mmol) was added 1 mL of degassed DMF and 0.5 mL of degassed H2O. To this was added CH3CO2H (0.0103 mL, 0.180 mmol). The mixture was stirred for 72 h. LC-MS indicated the formation of the desired product. The mixture was purified by preparative HPLC (Sunfire C18 150×30 mm; 20-77% H2O / acetonitrile with 0.5% AcOH modifier; run 15 min; flow rate: 30 ml / min; wavelength = 217 nM) to give the desired conjugate (17.0 mg, yield: 59.1%).

[0462] Example 6: Synthesis of Compound 6

[0463]

[0464] To a vial containing Pv2-SPyr (Intermediate II-2; 27.0 mg, 6.55e-6 mol) and (2S)-2-[methyl-(4-methyl-4-thio-pentanoyl)amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10,12,14(26),16,18-pentaen-6-yl ester] (7.67 mg, 0.01 mmol) was added 1 mL of degassed DMF and 0.5 mL of degassed H2O. To this was added CH3CO2H (0.015 mL, 0.262 mmol). The mixture was stirred for 72 h. LC-MS indicated the formation of the desired product. The mixture was purified by preparative HPLC (Sunfire C18 150×30 mm; 20-80% H2O / acetonitrile with 0.5% AcOH modifier; run 16 min; flow rate: 30 ml / min; wavelength = 217 nM) to give the desired conjugate (11.4 g, yield: 36.6%).

[0465] Example 9. Synthesis of Compound 9

[0466]

[0467] To a vial containing Pv2 (25.0 mg, 0.006 mol; as a free-flowing solid) and (2S)-2-[3-[2-[3-(2,5-dioxopyrrol-1-yl)propionylamino]ethyldisulfanyl]propionyl-methylamino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10(26),11,13,16,18-pentaen-6-yl ester] (Intermediate X-1; 0.00877 g, 0.01 mmol) was added 1 mL of The mixture was heterogeneous. 0.5 mL of CH 3 CN, 0.5 mL of H 2 O, and 0.5 mL of MeOH were added. Homogeneity was not achieved. The mixture was stirred rigorously for 72 h. LC-MS indicated the formation of the desired product. The mixture was purified by preparative HPLC (Sunfire C18 150×30 mm; 45-61% H 2 O / acetonitrile containing 0.05% TFA modifier; run 13 min; flow rate: 30 ml / min; wavelength = 217 nM) to give the desired conjugate (21.1 mg, yield: 70.0%).

[0468] Compounds 1, 3, and 4 were synthesized similarly to compound 2 using Pv1, Pv3, and Pv4, respectively. Compounds 5, 7, and 8 were synthesized similarly to compound 6 using intermediates II-1, II-3, and II-6, respectively.

[0469] Table 4. Example compounds

[0470]

[0471]

[0472]

[0473]

[0474] Example 5: Detailed Synthesis of Compound 5

[0475] To a 3 mL flask containing Pv1 (50.0 mg, 1.48e-5 mol) and (2S)-2-[methyl-(4-methyl-4-thio-pentanoyl)amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa-10,12,14(26),16,18-pentaen-6-yl ester] (0.0150 g, 1.92e-5 mol) was added. N-Methylmorpholine (0.0600 mL, 0.000546 mol) was added to 2:1 CH3CN / H2O. The mixture was stirred for 36 h. LC-MS analysis indicated the formation of the desired material. The mixture was purified by Gilson preparative HPLC (Sunfire C18 30×150 mm; 20-80% CH3CN / H2O with 0.05% TFA; run time 16 min; 13.5 min) to give the desired conjugate. The mixture was purified by Gilson preparative HPLC (Sunfire C18 30×150 mm; 20-72% CH3CN / H2O with 0.05% TFA; run time 15 min; 12.5 min; retention time: 6.847 min) to give compound 5 (0.0322 g, 7.94e-6 mol, yield: 53.8%). ESI (m / z=3): 1352.8.

[0476] Example 5a: Alternative Synthesis of Compound 5

[0477] Step 1. Preparation of Pv1-S-pyridyl

[0478] Peptide Pv1 and 2,2'-dipyridyl disulfide were dissolved in MeOH and stirred overnight. LC-MS indicated the formation of the desired product. The reaction mixture was concentrated and the residue was dissolved in DMSO and purified by reverse phase column chromatography (40-75% ACN / H2O (0.5% AcOH), 15 min) to give 212 mg of the desired product.

[0479] Step 2. Preparation of compound 5

[0480] To a vial containing Pv1-SPyr (25.0 mg, 736e-6 mol) and (2S)-2-[methyl-(4-methyl-4-thio-pentanoyl)amino]propanoic acid [(1S,2R,3S,6S,16E,18E,20R,21S)-11-chloro-21-hydroxy-12,20-dimethoxy-2,5,9,16-tetramethyl-8,23-dioxo-4,24-dioxa-9,22-diazatetracyclo[19.3.1.110,14.03,5]hexacosa10,12,14(26),16,18-pentaen-6-yl ester] (0.00864 g, 1.11e-5 mol) was added 1 mL of degassed DMF and 0.5 mL of degassed H2O. CH3CO2H (0.017 mL, 0.000295 mol) was added. The mixture was stirred for 72 h. LC-MS indicated the formation of the desired product. The mixture was purified by Gilson preparative HPLC (Sunfire C18 30x150 mm; 20-80 CH3CN / H2O containing 0.5% AcOH; run 16 min; 12.9 min) to give compound 5 (0.00750 g, 1.85e-6 mol, yield: 25.1%).

[0481] Example 10: Synthesis of Compound 10

[0482]

[0483] Step 1. N-(4-((2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0484]

[0485] 180mg DM4 (0.23mmol) and 57mg bromoacetic acid N-hydroxysuccinimide ester (0.24mmol) are dissolved in DMF (4.6mL) and cooled in an ice-water bath. 36.2 μL DBU (0.24mmol) are added once and the mixture is warmed to RT. At this time, LC / MS indicates a conversion rate close to 95% and the reactant is quenched by adding 0.1mL AcOH. The crude reaction mixture is directly loaded onto a 50g C18Aq post and purified via a standard 10-100%B gradient (A: water containing 0.05% AcOH; B: water containing 0.05% AcOH). The fraction containing the product is freeze-dried to obtain 160mg of the product (77% yield). HPLC purity at 254nm: 96%. Retention time: 2.83min (method F). LCMS: 935.4MH + .

[0486] Step 2. N-(4-((2-((2-aminoethyl)amino)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine 1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1+-hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0487]

[0488] 25 mg of N-(4-((2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine 1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzyl-10,12-dien-4-yl ester (0.027 mmol) and 36 mg of N1-((4-methoxyphenyl)diphenylmethyl)ethane-1,2-diamine (0.11 mmol, 4 eq) were dissolved in dioxane (1 mL). After 3 h, the reaction appeared to be complete according to LC / MS. The mixture was concentrated to dryness and dissolved in 80% AcOH / water (2 mL). LC / MS showed that the deprotection of the intermediate was complete and the mixture was directly lyophilized. The residue was dissolved in DMSO (1 mL) and loaded onto a 15.5 g C18Aq column and purified via a standard 5-100% B gradient (A: water containing 0.05% AcOH; B: water containing 0.05% AcOH). The product-containing fractions were lyophilized to give 18 mg of product. HPLC purity at 254 nm: 95%. Retention time: 2.17 min (Method F). LCMS: 880.4 MH + .

[0489] Step 3. (2S,18S)-2,3,7,7-Tetramethyl-4,10,15-trioxo-18-(pyridin-2-yldisulfanyl)-16-oxa-8-thia-3,11,14-triazanonanodecanoic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0490]

[0491] N-(4-((2-((2-aminoethyl)amino)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3,2,7,10-tetramethyl-1 2 A solution of 6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarbamide-10,12-dien-4-yl ester (14 mg, 0.016 mmol) in DMF (0.2 mL) was added to solid (S)-4-nitrophenyl (2-(pyridin-2-yldisulfanyl)propyl) carbonate (6.6 mg, 0.018 mmol). Catalytic HOAt and DIEA (10 mL, 0.057 mmol) were added to the resulting solution and stirred at room temperature for 3 hours. The solution was neutralized with acetic acid (10 mL) and applied to a reverse phase column (RediSEP C18 (15.5 g)) and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 18 mg (85% yield) of the title product. HPLC purity at 254 nm: 99%. Retention time: 2.85 min (Method F). LCMS: 1129.4 M Na + .

[0492] Step 4. Synthesis of compound 10

[0493] (2S,18S)-2,3,7,7-tetramethyl-4,10,15-trioxo-18-(pyridin-2-yldisulfanyl)-16-oxa-8-thia-3,11,14-triazanonadecanoic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2A solution of 1,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-diene-4-yl ester (17.7 mg, 0.00857 mmol) in DMF (1 mL) was treated with sodium bicarbonate (1.8 mg, 0.0214 mmol) and water (50 mL). The resulting solution was treated with the peptide, Pv1 (31.5 mg, 0.0899 mmol) and stirred at room temperature for 3 hours, then applied to a reverse phase column (RediSep C18 (15.5 g)) and eluted with a gradient of acetonitrile (30% to 70%) / water containing ammonium acetate (10 mM). The fractions were combined, frozen and lyophilized to give the product as a white solid, 18.7 mg (50%). HPLC purity at 254 nm: 99%. Retention time: 6.49 min (Method G) LCMS: 2138.0 (M+2H) / 2 + ,1425.3(M+3H) / 3+.

[0494] Example 11: Synthesis of Compound 11

[0495]

[0496] Step 1. N-(4-((4-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-4-oxobutyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0497]

[0498] DM4 (10 mg, 0.013 mmol) and 4-(2-pyridyldisulfanyl)butanoic acid succinimidyl ester (6 mg, 0.02 mmol) were mixed in DMF (0.26 mL). Triethylamine (0.015 mL) was added and the mixture was stirred for 2 h. 1-(2-aminoethyl)-1H-pyrrole-2,5-dione hydrochloride (5 mg, 0.026 mmol) was added and after 3 h, the mixture was directly loaded onto a RediSEPC18Aq (15.5 g) column and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 6 mg (40% yield) of the title product. HPLC purity at 254 nm: 92%. Retention time: 2.83 min (Method F). LCMS: 1020.4 MH + .

[0499] Step 2. Synthesis of compound 11

[0500] N-(4-((4-((2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethyl)amino)-4-oxobutyl)disulfanyl)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (6 mg, 0.006 mmol) and Pv1 peptide (22.3 mg, 0.006 mmol) were dissolved in DMF (0.12 mL) and treated with triethylamine (0.001 mL). After 30 minutes, the reaction mixture was directly loaded onto a RediSEP C8 (15.5 g) column and eluted with a gradient of acetonitrile (35% to 75%) in water containing TFA (0.05%) to give 16 mg (64% yield) of the title compound. HPLC purity at 254 nm: 98%. Retention time: 6.19 min (Method G). LCMS: 2150.2 (M+2H) / 2 + ,1433.3(M+3H) / 3 + .

[0501] Example 12. Synthesis of Compound 12

[0502]

[0503] Step 1. (S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-16,16,20,21-tetramethyl-10,19-dioxo-3,6-dioxa-14,15-dithia-9,20-diazadocosan-22-oic acid ((1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0504]

[0505] DM4 (20 mg, 0.026 mmol) and 4-(2-pyridyldisulfanyl)butanoic acid succinimidyl ester (12 mg, 0.04 mmol) were mixed in DMF (0.75 mL). Triethylamine (0.045 mL) was added and the mixture was stirred for 2 h. 1-(21-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrrole-2,5-dione hydrochloride (9 mg, 0.036 mmol) was added and after 3 h, the mixture was directly loaded onto a RediSEP C18Aq (15.5 g) column and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 17 mg (61% yield). HPLC purity at 254 nm: 99%. Retention time: 2.84 min (Method F). LCMS: 1108.4 MH + .

[0506] Step 2. Synthesis of compound 12.

[0507] (S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-16,16,20,21-tetramethyl-10,19-dioxo-3,6-dioxa-14,15-dithia-9,20-diazadocosan-22-oic acid (1 4 S,1 6 S,3 2 S,3 3S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (15 mg, 0.014 mmol) and Pv1 peptide (52 mg, 0.015 mmol) were dissolved in DMF (0.28 mL) and treated with triethylamine (0.006 mL). After 30 minutes, the reaction mixture was directly loaded onto a RediSEP C8 (15.5 g) column and eluted with a gradient of acetonitrile (35% to 60%) in water containing TFA (0.05%) to give 25 mg (34% yield). HPLC purity at 254 nm: 98%. Retention time: 6.26 min (Method G). LCMS: 2194.0 (M+2H) / 2 + ,1463.0(M+3H) / 3 + .

[0508] Example 13. Synthesis of Compound 13

[0509]

[0510] Step 1. (S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-13,13,17,18-tetramethyl-7,16-dioxo-3-oxa-11,12-dithia-6,17-diazodecanoic acid 1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0511]

[0512] DM4 (20 mg, 0.026 mmol) and 4-(2-pyridyldisulfanyl)butanoic acid succinimidyl ester (12 mg, 0.04 mmol) were mixed in DMF (0.75 mL). Triethylamine (0.045 mL) was added and the mixture was stirred for 2 h. 1-(2-(2-aminoethoxy)ethyl)-1H-pyrrole-2,5-dione hydrochloride (5 mg, 0.024 mmol) was added and after 3 h the mixture was directly loaded onto a RediSEP C18Aq (15.5 g) column and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 13 mg (41% yield) of the title product. HPLC purity at 254 nm: 94%. Retention time: 2.85 min (Method F). LCMS: 1064.4 MH + .

[0513] Step 2. Synthesis of compound 13

[0514] (S)-1-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)-16,16,20,21-tetramethyl-10,19-dioxo-3,6-dioxa-14,15-dithia-9,20-diazadocosan-22-oic acid (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy- A solution of 3,3,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxiran-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (18 mg, 0.017 mmol) and Pv1 peptide (63 mg, 0.019 mmol) were dissolved in DMF (0.34 mL) and treated with triethylamine (0.007 mL). After 30 minutes, the reaction mixture was directly loaded onto a RediSEP C8 (15.5 g) column and eluted with a gradient of acetonitrile (35% to 60%) in water containing TFA (0.05%) to give 25 mg (34% yield) of the title compound. HPLC purity at 254 nm: 99%. Retention time: 6.24 min (Method G). LCMS: 2172.0 (M+2H) / 2 + ,1448.7(M+3H) / 3 + .

[0515] Example 14. Synthesis of Compound 14

[0516]

[0517] Step 1. (S)-9,9,13,14-tetramethyl-1,6,12-trioxo-1-(((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl)oxy)-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0518]

[0519] N-(4-((2-((2-aminoethyl)amino)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 1,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarbimide-10,12-dien-4-yl ester (14 mg, 0.016 mmol; Example 10, Step 2) in DMF (0.2 mL) was added to solid 4-nitrophenyl carbonate ((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl ester) (7.2 mg, 0.018 mmol). Catalytic HOAt and DIEA (10 mL, 0.057 mmol) were added to the resulting solution and stirred at room temperature for 3 hours. The solution was neutralized with acetic acid (10 mL) and applied to a reverse phase column RediSEP C18 (15.5 g) and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 15 mg (80% yield) of the title compound. HPLC purity at 254 nm: 98%. Retention time: 3.05 min (Method F). LCMS: 1147.4 MH +.

[0520] Step 2. Synthesis of compound 14

[0521] (S)-9,9,13,14-tetramethyl-1,6,12-trioxo-1-(((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl)oxy)-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (17 mg, 0.015 mmol) and Pv1 peptide (47 mg, 0.013 mmol) were dissolved in DMF (0.34 mL) and treated with triethylamine (0.007 mL). After 30 minutes, the reaction mixture was directly loaded onto a RediSEP C8 (15.5 g) column and eluted with a gradient of acetonitrile (35% to 60%) in water containing TFA (0.05%) to give 28 mg (37% yield). HPLC purity at 254 nm: 99%. Retention time: 7.36 min (Method G). LCMS: 2158.0 (M+2H) / 2 + ,1439.0(M+3H) / 3 + .

[0522] Example 15. Synthesis of Compound 15

[0523]

[0524] Step 1. (S)-5,9,9,13,14-pentamethyl-6,12-dioxo-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0525]

[0526] 25 mg of N-(4-((2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzyltetracarbamide-10,12-dien-4-yl ester (0.027 mmol) and 40 mg of N1-((3-methoxyphenyl)benzhydryl)-N1,N2-dimethylethane-1,2-diamine (0.11 mmol, 4 eq) were dissolved in dioxane (1 mL). After 3 h, the reaction appeared to be complete according to LC / MS. 0.05 mL of TFA was added and the mixture was loaded onto a 15.5 g C18Aq column and purified via a standard 5-100% B gradient (A: water with 0.05% TFA; B: ACN with 0.05% TFA). The fractions containing the product were lyophilized to give 22 mg of product (75% yield). HPLC purity at 254 nm: 98%. Retention time: 2.25 min (Method F). LCMS: 908.4 MH + .

[0527] Step 2. (S)-2,5,9,9,13,14-Hexamethyl-1,6,12-trioxo-1-(((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl)oxy)-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3,2,7,10-tetramethyl-1 2 ,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-benzacyclotetracarbamate-10,12-dien-4-yl ester

[0528]

[0529] (S)-5,9,9,13,14-pentamethyl-6,12-dioxo-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6 -Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 1,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarbamide-10,12-dien-4-yl ester (15 mg, 0.016 mmol) in DMF (0.2 mL) was added to solid 4-nitrophenyl carbonate ((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl ester) (6.6 mg, 0.018 mmol). Catalytic HOAt and DIEA (10 mL, 0.057 mmol) were added to the resulting solution and stirred at room temperature for 3 hours. The solution was neutralized with acetic acid (10 mL) and applied to a reverse phase column (RediSEP C18 (15.5 g)) and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%) to give 16 mg (82% yield) of the title product. HPLC purity at 254 nm: 97%. Retention time: 3.36 min (Method F). LCMS: 1175.5 MH + .

[0530] Step 3. Synthesis of compound 15

[0531] (S)-2,5,9,9,13,14-hexamethyl-1,6,12-trioxo-1-(((1S,2S)-2-(pyridin-2-yldisulfanyl)cyclohexyl)oxy)-8-thia-2,5,13-triazapentadecan-15-oic acid (1 4 S,1 6 S,3 2 S,3 3 S,2R,4S,10E,12E,14R)-8 6-Chloro-1 4 -Hydroxy-8 5 ,14-dimethoxy-3 3 ,2,7,10-tetramethyl-1 2 A solution of 6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (16 mg, 0.014 mmol) and Pv1 peptide (52 mg, 0.015 mmol) were dissolved in DMF (0.28 mL) and treated with triethylamine (0.008 mL). After 30 minutes, the reaction mixture was directly loaded onto a RediSEP C8 (15.5 g) column and eluted with a gradient of acetonitrile (35% to 60%) in water containing TFA (0.05%) to give 32 mg (44% yield) of the title compound. HPLC purity at 254 nm: 99%. Retention time: 6.86 min (Method G). LCMS: 2172.0 (M+2H) / 2 + ,1448.4(M+3H) / 3 + .

[0532] Example 16. Synthesis of Compound 16

[0533]

[0534] Step 1. (4-((5-Nitropyridin-2-yl)disulfanyl)phenyl)methanol

[0535]

[0536] A solution of (4-mercaptophenyl)methanol (0.74 g, 4.83 mmol) in THF (10 mL) was treated with 5-nitro-2-((4-nitrophenyl)disulfanyl)pyridine (1.0 g, 3.23 mmol). The resulting suspension was stirred at room temperature for 2 hours, and the solvent was evaporated in vacuo. The residue was dissolved in DCM and applied to a RediSep silica gel column and eluted with a gradient of ethyl acetate (10% to 60%) / hexane to give the product (0.499 g, 52% yield). HPLC purity at 254 nm: 90%. Retention time: 2.72 min (Method F). MS data, 295.1 (M+H) + . 1 H NMR (DMSO-d6) d 9.18 (s, 1H), 8.58 (doublet of doublet, 1H), 8.02 (d, 1H), 7.56 (d, 2H), 7.34 (d, 2H), 5.24 (t, 1H) and 4.47 (d, 2H).

[0537] Step 2. 4-Nitrophenyl Carbonate (4-((5-nitropyridin-2-yl)disulfanyl)benzyl ester)

[0538]

[0539] A solution of 4-nitrophenyl chloroformate (255 mg, 1.26 mmol) in THF (5 mL) was cooled in an ice bath and treated over approximately 15 minutes with (4-((5-nitropyridin-2-yl)disulfanyl)phenyl)methanol (220 mg, 0.748 mmol), triethylamine (0.7 mL, 5.03 mmol), and 4-dimethylaminopyridine (45 mg, 0.368 mmol) in THF (5 mL). The ice bath was removed, and the solution was stirred at room temperature for one hour and stored in the freezer overnight. The solvent was evaporated in vacuo, and the residue was dissolved in DCM, applied to a RediSep silica gel column (12 g), and eluted with a gradient of ethyl acetate (2% to 100%) in hexanes. The product was further purified by reverse phase chromatography on a RediSep C18 cartridge (50 g) eluting with a gradient of acetonitrile (30% to 95%) in water containing 0.05% acetic acid to afford 55 mg (16%) of the product. HPLC purity at 254 nm: >99%. Retention time: 3.77 min (Method F). MS data, 460.7 (M+H). + . 1 HNMR (CDCl3) d 9.29 (d, 1H), 8.39 (doublet of doublet, 1H), 8.28 (doublet of doublet, 2H), 7.83 (doublet of doublet, 1H), 7.55 (doublet of doublet, 2H), 7.44 (doublet of doublet), 7.36 (doublet of doublet, 2H) and 5.26 (d, 2H).

[0540] Step 3. (S)-11,11,15,16-tetramethyl-1-(4-((5-nitropyridin-2-yl)disulfanyl)phenyl)-3,8,14-trioxo-2-oxa-10-thia-4,7,15-triazaheptadecan-17-oic acid (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxiran-8(1,3)-benzyl-10,12-dien-4-yl ester

[0541]

[0542] N-(4-((2-((2-aminoethyl)amino)-2-oxoethyl)thio)-4-methylpentanoyl)-N-methyl-L-alanine (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-33,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4 A solution of 5-nitrophenyl-3-nitroaniline-8-(1,3)-oxirane-10,12-dien-4-yl ester (15 mg, 0.017 mmol; Example 10, Step 2) in DMF (1 mL) was added to solid 4-nitrophenyl (4-((5-nitropyridin-2-yl)disulfanyl)benzyl carbonate) (26 mg, 0.0566 mmol). Catalytic HOAt and DIEA (10 mL, 0.057 mmol) were added to the resulting solution and stirred at room temperature for 3 hours. The solution was neutralized with acetic acid (7 mL, 0.122 mmol) and applied to a reverse phase column (RediSEP C18 (15.5 g)) and eluted with a gradient of acetonitrile (30% to 95%) / water containing acetic acid (0.05%). Further purification was performed on a silica gel column (RediSep (4 g)) using a gradient of methanol (0.2% to 6%) / DCM as eluent to give the title product (10.3 mg, 50% yield). HPLC purity at 254 nm: >99%. Retention time: 3.16 min (Method F). MS data, 1182.3 (M+H-H2O) + ,1201.3(M+H) + ,1222.3(M+Na) + .

[0543] Step 4. Synthesis of compound 16

[0544] (S)-11,11,15,16-tetramethyl-1-(4-((5-nitropyridin-2-yl)disulfanyl)phenyl)-3,8,14-trioxo-2-oxa-10-thia-4,7,15-triazaheptadecan-17-oic acid (14S,16S,32S,33S,2R,4S,10E,12E,14R)-86-chloro-14-hydroxy-85,14-dimethoxy-3 A solution of 3,2,7,10-tetramethyl-12,6-dioxo-7-aza-1(6,4)-azacyclohexane-3(2,3)-oxirane-8(1,3)-phencyclotetracarpine-10,12-dien-4-yl ester (10.3 mg, 0.00857 mmol) in DMF (1 mL) was treated with sodium bicarbonate (1.8 mg, 0.0214 mmol) and water (50 mL). The resulting solution was treated with the peptide, Pv1 (31.5 mg, 0.0899 mmol) and stirred at room temperature for 3 hours before being applied to a reverse phase column (RediSep C18 (15.5 g)) and eluted with a gradient of acetonitrile (30% to 70%) / water containing ammonium acetate (10 mM). The fractions were combined, frozen, and lyophilized to give the product as a white solid, 18.7 mg (50%). HPLC purity at 254 nm: 99%. Retention time: 6.63 min (Method G). MS data, 2162.4 (M+2H) / 2 + ,1441.8(M+3H) / 3 + ,1082.6(M+4H) / 4 + ,1435.7(M+3H-H2O) / 3 + .

[0545] Example A. Growth Delay Assay

[0546] In the growth medium containing 10% FBS, cells are seeded in 96-well black wall transparent bottom plates (Griener), DLD-1WT cells are seeded at 2500 cells / well, FaDu and HeLa cells are seeded at 5000 cells / well, and HCT116 is seeded at 3000 cells / well. Before returning to 37°C, 5% CO2 incubator, cells are allowed to adhere at room temperature for 60 minutes. After 24 hours, the culture medium is removed and replaced with fresh growth medium containing different drug concentrations. Each drug concentration is added in triplicate. The control without drug treatment contains only growth medium. The cells are returned to the incubator. Ninety-six hours after adding the drug, the cells are fixed with 4% paraformaldehyde for 20 minutes and stained with 1 μg / mL of Hoechst. The plate is imaged on a Cytation 5 automatic imager (BioTek) and the cells are counted using CellProfiler (http: / / cellprofiler.org). The cell growth delay percentage is calculated and the data are plotted using GraphPad Prism

[0547] Table 5: Growth Delay Assay Data

[0548]

[0549] NC* = Not Calculated

[0550] Example B: Effects on in vitro tubulin polymerization

[0551] Fluorescence-based tubulin polymerization assay (Cytoskeleton catalog number BK011P) was performed to quantify the effects of unconjugated DM4 and compound 5 on in vitro tubulin polymerization. DM4 and compound 5 were prepared as DMSO containing 10mM stock solutions and then diluted to 200, 50, and 5 μM at 10X in ultrapure distilled water, with a final DMSO concentration of 0.2%. The kit reagents were quickly thawed and then kept cold on ice to prevent premature polymerization. The tubulin reaction mixture was prepared on ice by mixing purified porcine brain tubulin, GTP, and glycerol buffer in 1X kit buffer to a final concentration of 2 mg / mL tubulin, 1 mM GTP, and 15% glycerol. At 37°C, 5 μL of DM4, compound 5, or DMSO control was added to a pre-warmed black half-well reaction plate for no more than 1 minute to warm but not allow evaporation. 50 μL of tubulin reaction mixture was quickly added to each well and immediately placed in a pre-warmed Cytation 5 imaging reader (BioTek). Kinetic readings were performed at 360 excitation / 450 emission for 2 hours at 37°C, with readings taken every 2.5 minutes to track the increase in fluorescence due to incorporation of the fluorescent reporter into microtubules as polymerization occurred.

[0552] Figure 1 Graph showing the effect of free DM4 and compound 5 on in vitro β-tubulin polymerization (in terms of relative fluorescence units) at 0.5 μM, 5 μM and 20 μM.

[0553] Example C: Kinetic Analysis of Conjugate Binding

[0554] Binding experiments were performed using a Biacore S200 instrument. A series S sensor chip with pre-fixed streptavidin was adjusted with 50mM NaOH containing 1M NaCl. Biotin-labeled human tubulin derived from HeLa cells was fixed to the sensor chip at a concentration of 125μg / mL at a flow rate of 10μl / min in HBS-P+ buffer. Final 3000RU (reaction units) of protein were directly fixed to the chip. After tubulin was fixed, the sensor chip was washed with 50% isopropanol, 50mM NaOH and 1M NaCl, and then balanced in assay buffer for 4 hours. A streptavidin-biotin capture blank (reference FC) was used to monitor nonspecific binding.

[0555] To collect kinetic binding data, compound 5 diluted in assay buffer was injected onto a flow cell at concentrations ranging from 100 μM to 0.048 μM and from 50 μM to 0.024 μM at a flow rate of 60 μL / min and at a temperature of 25°C. The complex was allowed to dissociate for 60 seconds. The binding of the compound to tubulin was monitored in real time to obtain the association (Ka) and dissociation (Koff) rates. The affinity constant (KD) was calculated by steady-state kinetics.

[0556] Figure 2 Depicted is a kinetic analysis of compound 5 binding to β-tubulin in vitro, as determined by Biacore surface plasmon resonance. Compound 5 is able to bind to β-tubulin with a similar KD to free DM4 (3.55 μM) and slower association / dissociation rates relative to free DM4.

[0557] Example D: Efficacy of Compound 5 in a mouse colorectal cancer model

[0558] Six-week-old female athymic naked Fox nu Mice were obtained from Taconic Labs (Cat. No. NCRNU-F) and housed five per cage on Alpha-Dri bedding in a disposable cage system. Human HCT116 cells derived from colorectal cancer were diluted 1:1 in Matrigel without phenol red and plated at 2.5 × 10 6 The xenografts were implanted subcutaneously into the left flank of each mouse at a density of 100 cells / 100 μL. 3The mice were randomized into groups and treated as detailed in the table below when the mean volume of the mice was ≥ 100 μg / kg. Mice were administered intraperitoneal (IP) doses of vehicle or 0.21, 0.29, 0.35, 0.42 μmol / kg Compound 5 (equivalent to 1.1, 1.4, 1.7 or 2 mg / kg Compound 5) or 0.42 μmol / kg unconjugated DM4 (equivalent to 0.33 mg / kg unconjugated DM4). Doses were prepared by diluting 0.1 mg / μL of 5% mannitol containing DMSO stock in citrate buffer and QDX4 was administered at a volume of 12 mL / kg (300 μL / 25 g mouse) with a two-day interval between the second and third doses. Xenograft tumors were measured by calipers and the volume was calculated using the ellipsoid volume equation: Volume = π / 6×(length)×(width) 2 The body weight of the animals was measured at the same time as the tumor volume was assessed. Animals that died or had tumors larger than 2000 mm 3 Patients were removed from the study if they had a weight loss of >20%. Survival was assessed based on death or removal from the study using Kaplan-Meier analysis.

[0559] Figure 3A Graph showing mean tumor volume of nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5.

[0560] Figure 3B Shown are the percentage changes in body weight relative to day 0 in nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5.

[0561] Figure 4 Figure 5. Kaplan-Meier plots of nude mice bearing HCT116 colorectal flank tumors administered with DM4 or Compound 5. Animals died due to tumors exceeding 2000 mm in size. 3 Or removed from the study due to weight loss of more than 20%. During the post-dose period, free DM4 induced spontaneous death in half of the animals in the DM4 group. Figure 4 As shown in , compound 5 safely delivers an amount of DM4 in vivo that would otherwise cause systemic toxicity and death when administered as free DM4.

[0562] Example E: Effect of Compound 6 on Lung Metastasis in Mouse Lung Cancer Model

[0563] Mouse 4T1-iRFP cancer cells derived from mouse breast cancer and transfected with near-infrared fluorescent protein (iRFP) were cultured as a monolayer at 37°C in a humidified atmosphere of 5% CO2. The cells were passaged one to three days before implantation and the culture medium was replaced every 2-3 days as needed to maintain cell viability. Cells were not allowed to exceed 80% confluence. On the day of implantation, the cells were trypsinized, washed with complete culture medium and pelleted by centrifugation at 1200 rpm for 5 minutes. The supernatant was decanted and the cells were washed three times with sterile PBS and pelleted by centrifugation. During the final centrifugation, viability was determined using trypan blue exclusion. The cells were resuspended in a final concentration of 5 × 10 5 100 μL of sterile PBS. Use a 27-gauge needle to draw the cells into a sterile 1cc tuberculin syringe. Remove air bubbles and drain the excess cell mixture back into the conical tube, leaving 100 μL of injection volume in each syringe. Inject 100 μL of cells directly into six-week-old female athymic naked Fox n nu The 5-well plate was injected into the medial tail vein of mice (Taconic Labs catalog number NCRNU-F).

[0564] Three days after cell injection, mice were administered intraperitoneal doses of vehicle or 2.5 mg / kg Compound 6 once daily for two days, followed by two days of no treatment, followed by a single dose of Compound 6, for a total of three doses of Compound 6. Eleven days after injection, mice were euthanized and lungs were removed for imaging using a LI-COR PEARL Trilogy small animal imager to visualize and quantify lung metastases and assess the effects of compounds on tumor growth.

[0565] Figure 5A Depicted are abdominal views and extracted lungs of nude mice inoculated with 4T1-RFP fluorescent cells via tail vein injection and imaged 11 days post-inoculation and after 3 doses of vehicle or compound 6.

[0566] Figure 5B Graph depicting fluorescent signals from extracted lungs of mice vaccinated with 4T1-RFP after 3 doses of vehicle or compound 6.

[0567] In addition to those modifications described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to be within the scope of the appended claims. Each reference cited in this application, including but not limited to all patents, patent applications, and publications, is hereby incorporated by reference in its entirety. Sequence Listing <110> Cyberxa 3, Inc. <120> Peptide conjugates of microtubule targeting agents as therapeutic agents <130> 43236-0008WO1 <150> 62 / 872,638 <151> 2019-07-10 <150> 63 / 041,324 <151> 2020-06-19 <160> 311 <170> PatentIn version 3.5 <210> 1 <211> 28 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <400> 1 Ala Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe Pro 1 5 10 15 Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp Cys Gly 20 25 <210> 2 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <400> 2 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Cys Gly 35 <210> 3 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <400> 3 Ala Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe Pro 1 5 10 15 Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp Asp Ala Asp Glu Cys Gly 20 25 30 <210> 4 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <220> <221> MISC_FEATURE <222> (1)..(1) <223> Acetylation <400> 4 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 5 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <400> 5 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys 35 <210> 6 <211> 39 <212> PRT <213> Unknown <220> <223> Description of unknown substance: pH-sensitive membrane polypeptide <400> 6 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Gly Thr Cys Gly 35 <210> 7 <211> 35 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 7 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 8 <211> 36 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 8 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 9 <211> 35 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 9 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 10 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 10 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 11 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 11 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 12 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 12 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 13 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 13 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 14 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 14 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 15 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 15 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 16 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 16 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Cys Thr 35 <210> 17 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 17 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asn Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asn Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 18 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 18 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Lys Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Lys Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 19 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 19 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asn Ala 20 25 30 Asn Gln Gly Thr 35 <210> 20 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 20 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Ala Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> twenty one <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> twenty one Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> twenty two <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> twenty two Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Ala Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> twenty three <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> twenty three Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> twenty four <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> twenty four Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 25 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 25 Ala Ala Glu Gln Asn Pro Ile Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Asp Leu Pro Leu Leu Leu Leu Asp Leu Leu Ala Leu Leu 20 25 30 Val Asp Ala Asp Glu Gly Thr 35 <210> 26 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 26 Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr Cys Gly 35 <210> 27 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 27 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Asp Leu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 28 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 28 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Leu Asp Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 29 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 29 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 30 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 30 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Trp Asp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 31 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 31 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Gly Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 32 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 32 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 33 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 33 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Gly Cys Thr 35 <210> 34 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 34 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 35 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 35 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 36 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 36 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Thr <210> 37 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 37 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 38 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 38 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Trp Asp Ala Asp 20 25 30 Glu Thr <210> 39 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 39 Ala Lys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 40 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 40 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 41 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 41 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Cys Gly 20 25 30 <210> 42 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 42 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Glu Cys Gly 20 25 30 <210> 43 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 43 Ala Lys Glu Asp Gln Asn Pro Tyr Trp Arg Ala Tyr Ala Asp Leu Phe 1 5 10 15 Thr Pro Leu Thr Leu Leu Asp Leu Leu Ala Leu Trp Asp Gly 20 25 30 <210> 44 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 44 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Arg Ala Tyr Ala Asp Leu Phe 1 5 10 15 Thr Pro Leu Thr Leu Leu Asp Leu Leu Ala Leu Trp Asp Gly 20 25 30 <210> 45 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 45 Ala Cys Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp 20 25 <210> 46 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 46 Thr Glu Asp Ala Asp Val Leu Leu Ala Leu Asp Leu Leu Leu Leu Pro 1 5 10 15 Thr Thr Phe Leu Trp Asp Ala Tyr Arg Ala Trp Tyr Pro Asn Gln Glu 20 25 30 Cys Ala <210> 47 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 47 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu 20 <210> 48 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 48 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Cys Leu 20 <210> 49 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 49 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu 20 <210> 50 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 50 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu 20 <210> 51 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 51 Lys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe Pro 1 5 10 15 Thr Thr Leu Ala Trp 20 <210> 52 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 52 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 53 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 53 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Asp 20 <210> 54 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 54 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Ala Glu Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 55 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 55 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Ala Glu Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Glu 20 <210> 56 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 56 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 57 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 57 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 58 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 58 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Thr 35 <210> 59 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 59 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 60 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 60 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Cys Thr 35 <210> 61 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 61 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 62 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 62 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 63 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 63 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 64 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 64 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Cys Thr 35 <210> 65 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 65 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Ala Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 66 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 66 Ala Cys Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Gly Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Gly Thr 35 <210> 67 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 67 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Thr <210> 68 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 68 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 69 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 69 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Trp Asp Ala Asp 20 25 30 Glu Thr <210> 70 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 70 Ala Lys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 71 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 71 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 72 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 72 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 73 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 73 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Glu Cys Gly 20 25 30 <210> 74 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 74 Ala Lys Glu Asp Gln Asn Pro Tyr Trp Arg Ala Tyr Ala Asp Leu Phe 1 5 10 15 Thr Pro Leu Thr Leu Leu Asp Leu Leu Ala Leu Trp Asp Gly 20 25 30 <210> 75 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 75 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Arg Ala Tyr Ala Asp Leu Phe 1 5 10 15 Thr Pro Leu Thr Leu Leu Asp Leu Leu Ala Leu Trp Asp Gly 20 25 30 <210> 76 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 76 Ala Lys Glu Asp Gln Asn Asp Pro Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Gly 20 25 30 <210> 77 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 77 Thr Glu Asp Ala Asp Val Leu Leu Ala Leu Asp Leu Leu Leu Leu Pro 1 5 10 15 Thr Thr Phe Leu Trp Asp Ala Tyr Arg Ala Trp Tyr Pro Asn Gln Glu 20 25 30 Cys Ala <210> 78 <211> 36 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 78 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 79 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 79 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu 20 <210> 80 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 80 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Cys Leu 20 <210> 81 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 81 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu 20 <210> 82 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 82 Ala Cys Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu 20 <210> 83 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 83 Ala Cys Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp 20 25 <210> 84 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 84 Ala Cys Glu Glu Gln Asn Pro Trp Arg Ala Tyr Leu Glu Leu Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu Leu Trp 20 25 <210> 85 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 85 Ala Cys Asp Asp Gln Asn Pro Trp Ala Arg Tyr Leu Asp Trp Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu 20 25 <210> 86 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 86 Cys Asp Asn Asn Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe Pro 1 5 10 15 Thr Asp Thr Leu Leu Leu Asp Trp 20 <210> 87 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 87 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 88 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 88 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Asp 20 <210> 89 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 89 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Ala Glu Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Glu 20 <210> 90 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 90 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 91 <211> twenty one <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 91 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Glu Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Trp 20 <210> 92 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 92 Lys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe Pro 1 5 10 15 Thr Thr Leu Trp 20 <210> 93 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 93 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 94 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 94 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 95 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 95 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 96 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 96 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 97 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 97 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 98 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 98 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 99 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 99 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Thr 35 <210> 100 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 100 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 101 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 101 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Asp Leu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 102 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 102 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Leu Asp Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 103 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 103 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Trp Asp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 104 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 104 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 105 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 105 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 106 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 106 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 107 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 107 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 108 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 108 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 109 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 109 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 110 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 110 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asn Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asn Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 111 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 111 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 112 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 112 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 113 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 113 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 114 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 114 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 115 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 115 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 116 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 116 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asn Ala 20 25 30 Asn Gln Gly Thr 35 <210> 117 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 117 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 118 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 118 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 119 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 119 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 120 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 120 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 121 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 121 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 122 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 122 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 123 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 123 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asn Ala 20 25 30 Asn Gln Gly Thr 35 <210> 124 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 124 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 125 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 125 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 126 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 126 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Lys Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Lys Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 127 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 127 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 128 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 128 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 129 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 129 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 130 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 130 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 131 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 131 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Asp Leu Pro Leu Leu Leu Leu Asp Leu Leu Ala Leu Leu Val 20 25 30 Asp Ala Asp Glu Gly Thr 35 <210> 132 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 132 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Leu Asp Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 133 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 133 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Asp Leu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 134 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 134 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Gly Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 135 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 135 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 136 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 136 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 137 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 137 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 138 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 138 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 139 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 139 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Ala Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 140 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 140 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 141 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37)..(37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 141 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 142 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Cys(Phalloidin) <400> 142 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 143 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Cys(Phalloidin) <400> 143 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 144 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 144 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 145 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 145 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Thr 35 <210> 146 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 146 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 147 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 147 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 148 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 148 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asn Ala 20 25 30 Asn Gln Gly Thr 35 <210> 149 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 149 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 150 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 150 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 151 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 151 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 152 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Cys(Phalloidin) <400> 152 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 153 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 153 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 154 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 154 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 155 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 155 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 156 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 156 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 157 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 157 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 158 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 158 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 159 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 159 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 160 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 160 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 161 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 161 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 162 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 162 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 163 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 163 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 164 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 164 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 165 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 165 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 166 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 166 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 167 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 167 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 168 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 168 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 169 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 169 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 170 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 170 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asn Ala 20 25 30 Asn Gln Gly Thr 35 <210> 171 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 171 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 172 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 172 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 173 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 173 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 174 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 174 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 175 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 175 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 176 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 176 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 177 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 177 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 178 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 178 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Cys Thr 35 <210> 179 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 179 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Cys Thr 35 <210> 180 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 180 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Cys Thr 35 <210> 181 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 181 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Gly Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 182 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 182 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Trp Asp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 183 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 183 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 184 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 184 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 185 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 185 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 186 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 186 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 187 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 187 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Leu Asp Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 188 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 188 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Asp Leu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 189 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 189 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Asp Leu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 190 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 190 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Leu Asp Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 191 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 191 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 192 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 192 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 193 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 193 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 194 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 194 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 195 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 195 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 196 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 196 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 197 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 197 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 198 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 198 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 199 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 199 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 200 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 200 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 201 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 201 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 202 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 202 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 203 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (4)..(4) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (5)..(5) <223> Cys(Phalloidin) <400> 203 Glu Gly Thr Lys Cys Gly 1 5 <210> 204 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 204 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 205 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 205 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 206 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Cys(Phalloidin) <400> 206 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 207 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 207 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 208 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 208 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 209 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 209 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Asp Leu Pro Leu Leu Leu Leu Asp Leu Leu Ala Leu Leu Val 20 25 30 Asp Ala Asp Glu Gly Thr 35 <210> 210 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 210 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 211 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 211 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 212 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 212 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Gln Asp Tyr Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 213 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 213 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Asp Ala Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 214 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 214 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 215 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 215 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 216 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 216 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Ala Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 217 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 217 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 218 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 218 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Glu Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 219 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (37) <223> Lys(Rhodamine) <220> <221> MOD_RES <222> (38)..(38) <223> Cys(Phalloidin) <400> 219 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 220 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 220 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Lys Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Lys Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 221 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 221 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asn Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asn Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 222 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 222 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Ala Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 223 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 223 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 224 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 224 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Phe 20 <210> 225 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 225 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 226 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 226 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Thr 35 <210> 227 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 227 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 228 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 228 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 229 <211> 39 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 229 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Lys Cys Gly 35 <210> 230 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 230 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Cys Thr 35 <210> 231 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 231 cctcttacct cagttaca 18 <210> 232 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 232 cctcttacct cagttaca 18 <210> 233 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 233 cctcttacct cagttaca 18 <210> 234 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 234 cctcttacct cagttaca 18 <210> 235 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 235 cctctgacct catttaca 18 <210> 236 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 236 cctcttacct cagttaca 18 <210> 237 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 237 cctctgacct catttaca 18 <210> 238 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic oligonucleotides <220> <221> misc_feature <222> (1)..(18) <223> Peptide nucleic acid <400> 238 cctcttacct cagttaca 18 <210> 239 <211> 38 <212> PRT <213> Unknown <220> <223> Description of unknown substance: pH-sensitive membrane polypeptide <400> 239 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 240 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 240 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Asp Trp Leu Phe Thr 1 5 10 15 Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Cys Gly 20 25 30 <210> 241 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 241 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Glu Cys Gly 20 25 30 <210> 242 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 242 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Cys Thr 35 <210> 243 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 243 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Thr 35 <210> 244 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 244 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 245 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 245 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 246 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 246 Ala Glu Asp Gln Asn Asp Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Gly 20 25 30 <210> 247 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 247 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Thr <210> 248 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 248 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Thr <210> 249 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 249 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Phe Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Trp Asp Ala Asp 20 25 30 Glu Thr <210> 250 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 250 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Cys Thr 35 <210> 251 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 251 Ala Glu Asp Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Cys 20 25 30 Gly Thr <210> 252 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 252 Ala Glu Asp Gln Asn Asp Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Glu 20 25 30 Cys Gly Thr 35 <210> 253 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 253 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 254 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 254 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Thr 35 <210> 255 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 255 Ala Lys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 256 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 256 Ala Lys Glu Asp Gln Asn Asp Pro Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Gly 20 25 30 <210> 257 <211> 35 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 257 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Cys 35 <210> 258 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 258 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Cys 20 25 30 <210> 259 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 259 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Glu Cys 20 25 30 <210> 260 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 260 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 261 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 261 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Gly 20 25 30 <210> 262 <211> 30 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 262 Ala Cys Glu Asp Gln Asn Pro Tyr Trp Arg Ala Tyr Ala Asp Leu Phe 1 5 10 15 Thr Pro Leu Thr Leu Leu Asp Leu Leu Ala Leu Trp Asp Gly 20 25 30 <210> 263 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 263 Ala Cys Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp 20 25 <210> 264 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 264 Ala Cys Glu Glu Gln Asn Pro Trp Arg Ala Tyr Leu Glu Leu Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu Leu Trp 20 25 <210> 265 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 265 Ala Cys Asp Asp Gln Asn Pro Trp Ala Arg Tyr Leu Asp Trp Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu 20 25 <210> 266 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 266 Cys Asp Asn Asn Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe Pro 1 5 10 15 Thr Asp Thr Leu Leu Leu Asp Trp 20 <210> 267 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 267 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 268 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 268 Cys Glu Glu Gln Gln Pro Trp Ala Gln Tyr Leu Glu Leu Leu Phe Pro 1 5 10 15 Thr Glu Thr Leu Leu Leu Glu Trp 20 <210> 269 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 269 Cys Glu Glu Gln Gln Pro Trp Arg Ala Tyr Leu Glu Leu Leu Phe Pro 1 5 10 15 Thr Glu Thr Leu Leu Leu Glu Trp 20 <210> 270 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 270 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Asp 20 <210> 271 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 271 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Ala Glu Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Glu 20 <210> 272 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 272 Ala Cys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 273 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 273 Ala Cys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Ala Glu Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 274 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 274 Thr Glu Asp Ala Asp Val Leu Leu Ala Leu Asp Leu Leu Leu Leu Pro 1 5 10 15 Thr Thr Phe Leu Trp Asp Ala Tyr Arg Ala Trp Tyr Pro Asn Gln Glu 20 25 30 Cys Ala <210> 275 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 275 Cys Asp Asp Asp Asp Asp Asn Pro Asn Tyr Trp Ala Arg Tyr Ala Asn 1 5 10 15 Trp Leu Phe Thr Thr Pro Leu Leu Leu Leu Asn Gly Ala Leu Leu Val 20 25 30 Glu Ala Glu Glu Thr 35 <210> 276 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 276 Cys Asp Asp Asp Asp Asp Asn Pro Asn Tyr Trp Ala Arg Tyr Ala Pro 1 5 10 15 Trp Leu Phe Thr Thr Pro Leu Leu Leu Leu Pro Gly Ala Leu Leu Val 20 25 30 Glu Ala Glu Glu Thr 35 <210> 277 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 277 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala Asp 20 25 30 Glu Gly Cys Thr 35 <210> 278 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 278 Ala Lys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly 35 <210> 279 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 279 Ala Cys Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asn Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asn Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr 35 <210> 280 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 280 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Thr 35 <210> 281 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 281 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Thr 35 <210> 282 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 282 Cys Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His 1 5 10 15 Trp Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val 20 25 30 Asp Ala Asp Glu Thr 35 <210> 283 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 283 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asp 20 25 30 Ala Asp Glu Gly Thr 35 <210> 284 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 284 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asp Glu Gly Thr 35 <210> 285 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 285 Asp Asp Asp Glu Asp Asn Pro Ile Tyr Trp Ala Arg Tyr Ala His Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu His Gly Ala Leu Leu Val Asn 20 25 30 Ala Asn Glu Gly Thr 35 <210> 286 <211> 32 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 286 Ala Lys Glu Asp Gln Asn Asp Pro Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Gly 20 25 30 <210> 287 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 287 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Ala Asp Trp Leu Phe 1 5 10 15 Thr Thr Pro Leu Leu Leu Leu Glu Leu Ala Leu Leu Val Cys Gly 20 25 30 <210> 288 <211> 28 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 288 Ala Lys Asp Asp Gln Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu Leu Trp Cys 20 25 <210> 289 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 289 Ala Cys Glu Glu Gln Asn Pro Trp Arg Ala Tyr Leu Glu Leu Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu Leu Trp 20 25 <210> 290 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 290 Ala Cys Asp Asp Gln Asn Pro Trp Ala Arg Tyr Leu Asp Trp Leu Phe 1 5 10 15 Pro Thr Asp Thr Leu Leu Leu Asp Leu 20 25 <210> 291 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 291 Cys Asp Asn Asn Asn Pro Trp Arg Ala Tyr Leu Asp Leu Leu Phe Pro 1 5 10 15 Thr Asp Thr Leu Leu Leu Asp Trp 20 <210> 292 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 292 Cys Glu Glu Gln Gln Pro Trp Ala Gln Tyr Leu Glu Leu Leu Phe Pro 1 5 10 15 Thr Glu Thr Leu Leu Leu Glu Trp 20 <210> 293 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 293 Cys Glu Glu Gln Gln Pro Trp Arg Ala Tyr Leu Glu Leu Leu Phe Pro 1 5 10 15 Thr Glu Thr Leu Leu Leu Glu Trp 20 <210> 294 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 294 Cys Asp Asp Asp Asp Asp Asn Pro Asn Tyr Trp Ala Arg Tyr Ala Asn 1 5 10 15 Trp Leu Phe Thr Thr Pro Leu Leu Leu Leu Asn Gly Ala Leu Leu Val 20 25 30 Glu Ala Glu Glu Thr 35 <210> 295 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 295 Cys Asp Asp Asp Asp Asp Asn Pro Asn Tyr Trp Ala Arg Tyr Ala Pro 1 5 10 15 Trp Leu Phe Thr Thr Pro Leu Leu Leu Leu Pro Gly Ala Leu Leu Val 20 25 30 Glu Ala Glu Glu 35 <210> 296 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 296 Ala Glu Gln Asn Pro Ile Tyr Phe Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 297 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 297 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Phe 20 <210> 298 <211> twenty two <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 298 Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Leu Leu Phe 1 5 10 15 Pro Thr Thr Leu Ala Trp 20 <210> 299 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 299 Lys Glu Asp Gln Asn Pro Trp Ala Arg Tyr Ala Asp Leu Leu Phe Pro 1 5 10 15 Thr Thr Leu Trp 20 <210> 300 <211> twenty four <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 300 Ala Cys Glu Glu Gln Asn Pro Gln Ala Glu Tyr Ala Glu Trp Leu Phe 1 5 10 15 Pro Thr Thr Leu Leu Leu Leu Glu 20 <210> 301 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 301 Ala Ala Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 302 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <400> 302 Ala Lys Glu Glu Gln Asn Pro Trp Ala Arg Tyr Leu Glu Trp Leu Phe 1 5 10 15 Pro Thr Glu Thr Leu Leu Leu Glu Leu 20 25 <210> 303 <211> 38 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 303 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly Gly 35 <210> 304 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (1)..(1) <223> Ala or Asp <220> <221> MOD_RES <222> (2)..(2) <223> Ala, Asp, or Cys <220> <221> MOD_RES <222> (4)..(4) <223> Gln or Asp <220> <221> MOD_RES <222> (11)..(11) <223> Arg or Gln <220> <221> MOD_RES <222> (12)..(12) <223> Tyr or Asp <220> <221> MOD_RES <222> (13)..(13) <223> Ala, Asp, or Tyr <220> <221> MOD_RES <222> (14)..(14) <223> Asp, Asn, Glu, His, Lys, Ala or Trp <220> <221> MOD_RES <222> (15)..(15) <223> Trp or Asp <220> <221> MOD_RES <222> (19)..(19) <223> Thr or Asp <220> <221> MOD_RES <222> (20)..(20) <223> Pro, Gly or Ala <220> <221> MOD_RES <222> (24)..(24) <223> Leu or Asp <220> <221> MOD_RES <222> (25)..(25) <223> Asp, Leu, Asn, Glu, His, Lys or Ala <220> <221> MOD_RES <222> (26)..(26) <223> Leu, Asp or Gly <220> <221> MOD_RES <222> (31)..(31) <223> Asp or Asn <220> <221> MOD_RES <222> (33)..(33) <223> Asp or Asn <220> <221> MOD_RES <222> (34)..(34) <223> Glu or Gln <220> <221> MOD_RES <222> (35)..(35) <223> Gly or Cys <220> <221> MOD_RES <222> (37)..(37) <223> Gly or Cys <400> 304 Xaa Xaa Glu Xaa Asn Pro Ile Tyr Trp Ala Xaa Xaa Xaa Xaa Xaa Leu 1 5 10 15 Phe Thr Xaa Xaa Leu Leu Leu Xaa Xaa Xaa Ala Leu Leu Val Xaa Ala 20 25 30 Xaa Xaa Xaa Thr Xaa Gly 35 <210> 305 <211> 42 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (1)..(1) <223> May or may not exist <220> <221> MOD_RES <222> (21)..(21) <223> May or may not exist <220> <221> MOD_RES <222> (29)..(29) <223> May or may not exist <220> <221> MOD_RES <222> (40)..(40) <223> May or may not exist <400> 305 Asp Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp 1 5 10 15 Leu Phe Thr Thr Leu Pro Leu Leu Leu Leu Asp Leu Leu Ala Leu Leu 20 25 30 Val Asp Ala Asp Glu Gly Thr Lys Gly Gly 35 40 <210> 306 <211> 38 <212> PRT <213> Unknown <220> <223> Description of unknown substance: Wild-type pH-sensitive membrane polypeptide <400> 306 Gly Gly Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Gly Gly 35 <210> 307 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (1)..(1) <223> Gly, Asp, or Ala <220> <221> MOD_RES <222> (2)..(2) <223> Gly, Asp, or Cys <220> <221> MOD_RES <222> (4)..(4) <223> Gln or Asp <220> <221> MOD_RES <222> (11)..(11) <223> Arg or Gln <220> <221> MOD_RES <222> (12)..(12) <223> Tyr or Asp <220> <221> MOD_RES <222> (13) <223> Ala, Asp, or Tyr <220> <221> MOD_RES <222> (14)..(14) <223> Asp, Asn, Glu, His, Lys, Ala or Trp <220> <221> MOD_RES <222> (15)..(15) <223> Trp or Asp <220> <221> MOD_RES <222> (19)..(19) <223> Thr or Asp <220> <221> MOD_RES <222> (20)..(20) <223> Pro, Gly or Ala <220> <221> MOD_RES <222> (24)..(24) <223> Leu or Asp <220> <221> MOD_RES <222> (25)..(25) <223> Asp, Leu, Asn, Glu, His, Lys, or Ala <220> <221> MOD_RES <222> (26) <223> Leu, Asp, or Gly <220> <221> MOD_RES <222> (31)..(31) <223> Asp or Asn <220> <221> MOD_RES <222> (33)..(33) <223> Asp or Asn <220> <221> MOD_RES <222> (34)..(34) <223> Glu or Gln <220> <221> MOD_RES <222> (35)..(35) <223> Gly or Cys <400> 307 Xaa Xaa Glu Xaa Asn Pro Ile Tyr Trp Ala Xaa Xaa Xaa Xaa Xaa Leu 1 5 10 15 Phe Thr Xaa Xaa Leu Leu Leu Xaa Xaa Xaa Ala Leu Leu Val Xaa Ala 20 25 30 Xaa Xaa Xaa Thr Gly Gly 35 <210> 308 <211> 44 <212> PRT <213> Artificial Sequence <220> <223> Description of artificial sequences: synthetic peptides <220> <221> MOD_RES <222> (1)..(1) <223> May or may not exist <220> <221> MOD_RES <222> (7)..(7) <223> May or may not exist <220> <221> MOD_RES <222> (22)..(22) <223> May or may not exist <220> <221> MOD_RES <222> (30)..(30) <223> May or may not exist <220> <221> MOD_RES <222> (40)..(40) <223> May or may not exist <220> <221> MOD_RES <222> (42)..(42) <223> Lys, Cys, or absent <400> 308 Asp Gly Gly Glu Gln Asn Asp Pro Ile Tyr Trp Ala Arg Tyr Ala Asp 1 5 10 15 Trp Leu Phe Thr Thr Leu Pro Leu Leu Leu Leu Asp Leu Leu Ala Leu 20 25 30 Leu Val Asp Ala Asp Glu Gly Cys Thr Xaa Gly Gly 35 40 <210> 309 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> Synthesize a peptide modified at residue 37 of the CYS peptide, wherein the SS-linker is attached to the nitrogen of the amino-phalloidin <400> 309 Ala Ala Glu Gln Asn Pro Ile Tyr Trp Ala Arg Tyr Ala Asp Trp Leu 1 5 10 15 Phe Thr Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Ala 20 25 30 Asp Glu Gly Thr Cys Gly 35 <210> 310 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Cys residue 30 was modified with an SS linker linked to 2-aminophalloidin, Lys residue 29 was modified with an alkyl linker linked to rhodamine, and alanine was modified with a COCH3 group. Residue 1. <400> 310 Ala Glu Asp Gln Asn Pro Tyr Trp Ala Arg Tyr Asp Trp Leu Phe Thr 1 5 10 15 Thr Pro Leu Leu Leu Leu Asp Leu Ala Leu Leu Val Asp Cys Gly 20 25 30 <210> 311 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> synthetic peptides <400> 311 Gly Leu Ala Gly Leu Ala Gly Leu Leu Gly Leu Glu Gly Leu Leu Gly 1 5 10 15 Leu Pro Leu Gly Leu Leu Glu Gly Leu Trp Leu Gly Leu Glu Leu Glu 20 25 30 Gly Asn

Claims

1. A compound of formula (I): or a pharmaceutically acceptable salt thereof, wherein: where R 1 is a peptide comprising at least one of the following sequences: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1; Pv1), AEQNPIYWARYADWLFTTTPLLLLDLALLVDADECG (SEQ ID NO. 2; Pv2), ADDQNPWRAYLDLLFPTDTLLLLDLLWDADECG (SEQ ID NO. 3; Pv3); and AAEQNPIYWWARYADWLFTTTPLLLLDLALLVDADEGTCG (SEQ ID No. 6; Pv6); where R 1 Via R 1 The cysteine ​​residue of is connected to L; R 2 Selected from the group consisting of: ； L is the following group: ； in R 3 、R 4 、R 5 and R 6 Each independently selected from H, C 1-4 Alkyl, C 1-4 Alkenyl, C 6-10 Aryl, C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 4-10 membered heterocycloalkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 , wherein the C 1-4 Alkyl, C 1-4 Alkenyl, C 6-10 Aryl and 5-10 membered heteroaryl are each optionally substituted with 1, 2 or 3 substituents independently selected from the group consisting of halo, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ; or R 3 and R 4 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ; or R 3 and R 5 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ; or R 4 and R 6 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ; or R 5 and R 6 Together with the carbon atom to which it is attached, it forms C 3-14 cycloalkyl or 4-14 membered heterocycloalkyl, each of which is optionally substituted by 1, 2 or 3 substituents independently selected from the group consisting of: C 1-4 Alkyl, halogen, CN, NO2, OR a1 SR a1 、C(O)R b1 、C(O)NR c1 R d1 、C(O)OR a1 、OC(O)R b1 、OC(O)NR c1 R d1 NR c1 R d1 NR c1 C(O)R b1 NR c1 C(O)OR a1 and NR c1 C(O)NR c1 R d1 ; A is H or C1-4 alkyl; and R a1 、R b1 、R c1 and R d1 Each independently selected from H, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Haloalkyl, OH, CN, NO2 and CO2CH3; wherein the C 1-6 Alkyl and C 2-6 The alkenyl groups are each optionally substituted with OH, CN, NO2 or CO2CH.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1; Pv1).

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is a peptide comprising at least the following sequence: AEQNPIYWARYADWLFTTTPLLLLDLALLVDADECG (SEQ ID NO. 2; Pv2).

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWDADECG (SEQ ID NO. 3; Pv3).

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 is a peptide comprising at least the following sequence: AAEQNPIYWWARYADWLFTTTPLLLLDLALLVDADEGTCG (SEQ ID NO. 6; Pv6).

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R 2 yes: 。 7. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R 2 yes: 。 8. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R 2 yes: 。 9. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R 2 yes: 。 10. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein L is: 。 11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein R 3 、R 4 、R 5 and R 6 Each independently selected from H and C 1-4 alkyl.

12. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein R 3 、R 4 、R 5 and R 6 Each is H.

13. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein A is H.

14. The compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, wherein A is CH3.

15. The compound of claim 1, wherein the compound is selected from the group consisting of: ； ;and ； or a pharmaceutically acceptable salt of any of the foregoing, wherein Pv1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1).

16. The compound according to claim 1, wherein the compound is: ， or a pharmaceutically acceptable salt thereof, wherein Pv1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1).

17. The compound according to claim 1, which is: ， or a pharmaceutically acceptable salt thereof, wherein Pv1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1).

18. The compound according to claim 1, which is: ， or a pharmaceutically acceptable salt thereof, wherein Pv1 is a peptide comprising at least the following sequence: ADDQNPWRAYLDLLFPTDTLLLLDLLWCG (SEQ ID NO. 1).

19. A pharmaceutical composition comprising the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof.

20. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating cancer in a patient, wherein the cancer is a solid tumor.

21. The method of claim 20, wherein the cancer is selected from the group consisting of bladder cancer, bone cancer, glioma, breast cancer, esophageal cancer, pancreatic cancer, gallbladder cancer, gastrointestinal tumors, head and neck cancer, kidney cancer, liver cancer, lung cancer, prostate cancer, skin cancer, testicular cancer, and uterine cancer.

22. The use according to claim 20, wherein the cancer is an epithelial cancer.

23. The use according to claim 20, wherein the cancer is selected from cervical cancer, endometrial cancer, Ewing's sarcoma, gastric cancer, intestinal cancer, Kaposi's sarcoma, laryngeal cancer, melanoma, renal clear cell carcinoma and thyroid cancer.

24. The use according to claim 23, wherein the cancer is colorectal cancer.

25. The use according to claim 24, wherein the cancer is selected from colon cancer and rectal cancer.

26. The use of claim 20, wherein the cancer is selected from lung cancer, colorectal cancer, and gastric cancer.

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