Cell-permeable (icp)-socs3 recombinant protein and uses thereof

Abstract

The present invention relates to providing improved cell-permeable (iCP)-SOCS3 recombinant protein and uses thereof. Preferably, the iCP-SOCS3 recombinant protein may be used as protein-based anti-solid tumor agent by utilizing the platform technology for macromolecule intracellular transduction.

1 . An improved Cell-Permeable (iCP)-SOCS3 recombinant protein, which comprises a SOCS3 protein and an advanced macromolecule transduction domain (aMTD) being composed of 9-13 amino acid sequences and having improved cell and/or tissue permeability,
 wherein the aMTD is fused to one end or both ends of the SOCS3 protein and has the following features of:   (a) being composed of 3 or more amino acids sequences selected from the group consisting of Ala, Val, Ile, Leu, and Pro;   (b) having proline as amino acid sequences corresponding to any one or more of positions 5 to 8, and 12 of its amino acid sequence; and   (c) having an instability index of 40-60; an aliphatic index of 180-220; and a grand average of hydropathy (GRAVY) of 2.1-2.6, as measured by Protparam.   
     
     
         2 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein one or more solubilization domain (SD)(s) are further fused to the end(s) of one or more of the SOCS3 protein and the aMTD. 
     
     
         3 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the aMTD is composed of 12 amino acid sequences and represented by the following general formula:    Here, X(s) independently refer to Alanine (A), Valine (V), Leucine (L) or Isoleucine (I); and Proline (P) can be positioned in one of U(s) (either 5′, 6′, 7′ or 8′); and the remaining U(s) are independently composed of A, V, L or I, P at the 12′ is Proline.   
     
     
         4 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the SOCS3 protein has an amino acid sequence of SEQ ID NO: 814. 
     
     
         5 . The iCP-SOCS3 recombinant protein according to  claim 4 , wherein the SOCS3 protein is encoded by a polynucleotide sequence of SEQ ID NO: 815. 
     
     
         6 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the SOCS3 protein further comprises a ligand selectively binding to a receptor of a cell, a tissue, or an organ. 
     
     
         7 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the aMTD has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-240 and 822. 
     
     
         8 . The iCP-SOCS3 recombinant protein according to  claim 7 , wherein the aMTD is encoded by a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 241-480 and 823. 
     
     
         9 . The iCP-SOCS3 recombinant protein according to  claim 2 , wherein the SD(s) have an amino acid sequence independently selected from the group consisting of SEQ ID NOs: 798, 799, 800, 801, 802, 803, and 804. 
     
     
         10 . The iCP-SOCS3 recombinant protein of  claim 9 , wherein the SD(s) are encoded by a polynucleotide sequence independently selected from the group consisting of SEQ ID NOs: 805, 806, 807, 808, 809, 810, and 811. 
     
     
         11 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the iCP-SOCS3 recombinant protein has a histidine-tag affinity domain additionally fused to one end thereof. 
     
     
         12 . The iCP-SOCS3 recombinant protein according to  claim 11 , wherein the histidine-tag affinity domain has an amino acid sequence of SEQ ID NO: 812. 
     
     
         13 . The iCP-SOCS3 recombinant protein of  claim 12 , wherein the histidine-tag affinity domain is encoded by a polynucleotide sequence of SEQ ID NO: 813. 
     
     
         14 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the fusion is formed via a peptide bond or a chemical bond. 
     
     
         15 . The iCP-SOCS3 recombinant protein according to  claim 1 , wherein the iCP-SOCS3 recombinant protein is used for the treating, preventing, or delaying the onset of, solid tumor. 
     
     
         16 . A polynucleotide sequence encoding the iCP-SOCS3 recombinant protein of  claim 1 . 
     
     
         17 . A recombinant expression vector comprising the polynucleotide sequence of  claim 16 . 
     
     
         18 . A transformant transformed with the recombinant expression vector of  claim 17 . 
     
     
         19 . A preparing method of the iCP-SOCS3 recombinant protein of  claim 1  comprising:
 preparing the recombinant expression vector of  claim 17 ; 
 preparing a transformant using the recombinant expression vector; 
 culturing the transformant; and 
 recovering the recombinant protein expressed by the culturing. 
 
     
     
         20 . A method of treating, preventing, or delaying the onset of, solid tumor in a subject comprising:
 identifying a subject in need of treating, preventing, or delaying the onset of, solid tumor; and   administering to the subject a therapeutically effective amount of the iCP-SOCS3 recombinant protein of  claim 1 .