Yap protein inhibiting polypeptide and application thereof

Abstract

Provided in the present invention are a YAP protein inhibiting polypeptide and application thereof. In particular, the present invention obtains a key binding site of YAP protein and TEAD, screens a polypeptide with best YAP inhibitory activity and modifies the polypeptide, such as adding a disulfide linkage, replacing an amino acid, removing and/or adding (for example, adding a cell-penetrating element), and finally screens and verifies the obtaining of a series of polypeptides with YAP protein activity inhibiting effect and good stability. Experiments show that the polypeptide of the present invention can effectively inhibit the binding activity between YAP protein and TEAD, thus providing good therapeutic effect on digestive tract tumors (especially liver cancer).

1 - 10 . (canceled) 
     
     
         11 . A polypeptide of formula V:
   X 0 -Z-X A -X E      wherein X 0  or X A  is not present, or is 1, 2, 3, or 4 amino acid residues;   X E  is not present or is a cell-penetrating element;   Z is a YAP inhibiting polypeptide having an intra-chain disulfide linkage;   Z has residues Z 1 , Z 2 , Z 3 , Z 4 , Z 5 , Z 6 , Z 7 , Z 8 , Z 9 , Z 10 , Z 11 , Z 12 , Z 13 , Z 14 , Z 15 , Z 16 , B 1 , B 2 ; and   Z 1 , Z 2 , Z 3 , Z 4 , Z 5 , Z 6 , Z 7 , Z 8 , Z 9 , Z 10 , Z 11 , Z 12 , Z 13 , Z 14 , Z 15 , and Z 16  has a sequence order represented by the formula Va: Z 1 -Z 2 -Z 3 -Z 4 -Z 5 -Z 6 -Z 7 -Z 8 -Z 9 -Z 10 -Z 11 -Z 12 -Z 13 -Z 14 -Z 15 -Z 16 ;
 wherein Z 1  is V, A or not present;
 Z 2  is P, A or not present; 
 Z 3  is M, or F; 
 Z 4  is R, K, A or not present; 
 Z 5  is L or Nle; 
 Z 6  is R or K; 
 Z 7  is K, R, Orn or Dab; 
 Z 8  is L or Nle; 
 Z 9  is P; 
 Z 10  is any natural or unnatural amino acid; 
 Z 11  is S or T; 
 Z 12  is F, L or chlorophenylalanine; 
 Z 13  is F, L, chlorophenylalanine or not present; 
 Z 14  is K, R, A, H, D or not present; 
 Z 15  is P or not present; 
 Z 16  is P or not present; 
 
   wherein the intra-chain disulfide linkage is formed by B 1  and B 2 , wherein B 1  is an amino acid residue inserted at a position between Z 1  and Z 2  when at least one of Z 1  and Z 2  is present or between Z 3  and Z 8  when both of Z 1  and Z 2  are not present, B 2  is an amino acid residue inserted at a position between Z 9  and Z 16  or after Z 16 .   
     
     
         12 . The polypeptide of  claim 11 , wherein the number of residues between B 1  and B 2  is 4, 5, 6, 7, 8, or 9 amino acids, wherein the number is counted without including B 1  and B 2 . 
     
     
         13 . The polypeptide of  claim 11 , wherein the polypeptide has the sequence selected from the group consisting of SEQ ID NOs.: 2-9, 11. 
     
     
         14 . The polypeptide of  claim 11 , wherein B1 and B2 are each independently Cys or Hcy. 
     
     
         15 . The polypeptide of  claim 11 , wherein X E  is selected from the group consisting of SEQ ID Nos.: 13, 14 and 15. 
     
     
         16 . The polypeptide of  claim 11 , wherein B 1  is located between Z 2  and Z 3 , between Z 3  and Z 4 , between Z 3  and Z 5  when Z 4  is not present, between Z 4  and Z 5 , or between Z 7  and Z 8 , and/or the second amino acid B 2  is located between Z 11  and Z 12 , between Z 12  and Z 13 , between Z 13  and Z 14 , after Z 14  when Z 15  and Z 16  are not present, after Z 15  when Z 16  is not present, or after Z 16 . 
     
     
         17 . The polypeptide of  claim 11 , wherein Z 10  is D, A, V, L, I or E. 
     
     
         18 . The polypeptide of  claim 11 , wherein the formula Va has at most 5 amino acid substitutions relative to SEQ ID NO: 1, the polypeptide of formula V retains≥60% biological activity of SEQ ID NO: 1, and the biological activity refers to an activity of inhibiting a binding between YAP and TEAD. 
     
     
         19 . A pharmaceutical composition comprising the polypeptide of  claim 11  or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 
     
     
         20 . A method of inhibiting YAP activity in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of  claim 19 . 
     
     
         21 . The method of  claim 20 , wherein the subject is in need of treating a tumor. 
     
     
         22 . The method of  claim 20 , wherein the subject is in need of treating a digestive tract tumor. 
     
     
         23 . The method of  claim 22 , wherein the digestive tract tumor comprises liver cancer, gastric cancer, colorectal cancer, gallbladder cancer, and pancreatic cancer. 
     
     
         24 . The method of  claim 23 , wherein the subject is in need of treating a liver cancer. 
     
     
         25 . The method of  claim 23 , wherein the subject is human.