Target-specific non-antibody protein and method for preparing the same

Patent metadata

Field	Value
Publication number	US-9296802-B2
Application number	US-201013264365-A
Country	US
Kind code	B2
Filing date	Apr 14, 2010
Priority date	Apr 15, 2009
Publication date	Mar 29, 2016
Grant date	Mar 29, 2016

Abstract

The present invention relates to a method for preparing a target-specific non-antibody protein, and more particularly, to a method for preparing a target-specific non-antibody protein comprising the steps of: selecting non-antibody proteins having a structural complementarity with the target site of a target protein in a non-antibody protein library; calculating a binding energy of the selected non-antibody protein and the target protein; selecting a non-antibody protein having a favorable binding energy among the selected non-antibody proteins; selecting amino acid residues having a high binding energy among the interfacial amino acid residues of the selected non-antibody protein and the target protein; and substituting the selected amino acid residues with the amino acid residues having a low binding energy. In addition, the present invention relates to a target-specific non-antibody specifically binding with EGFR (Epidermal Growth Factor Receptor) domain 2, which is prepared by the method, and a cancer therapeutic composition comprising the same.

First claim

What is claimed is:

1. A method for preparing an EGFR (Epidermal Growth Factor Receptor) domain 2 specific modified non-antibody protein comprising the steps of:
(a) selecting non-antibody proteins having a structural complementarity with the target site of EGFR domain 2 in a non-antibody protein library;
(b) calculating a binding energy of the selected non-antibody protein and EGFR domain 2;
(c) selecting a non-antibody protein having a favorable binding energy among the selected non-antibody proteins;
(d) selecting amino acid residue having high binding energy among the interfacial amino acid residues of the selected non-antibody protein and EGFR domain 2, wherein the selected amino acid residue having high binding energy is selected from 44Lys, 107Glu, 110Glu, 114Asn, and 118Asp; and
(e) substituting the amino acid residue selected in step (d) with the amino acid residue having low binding energy, wherein step (e) is performed by phage display and bio-panning.

2. The method according to claim 1 , wherein step (a) and step (b) are performed sequentially or at the same time.

3. The method according to claim 1 , wherein the non-antibody protein library of step (a) includes non-antibody proteins having a molecular weight of 10 to 40 kDa, selected from human proteins, and capable of forming any one or more of monomers, homodimers, and homotrimers.

4. The method according to claim 1 , wherein the library of step (a) is constructed by a method comprising the steps of:
(i) selecting proteins having a molecular weight of 10 to 40 kDa from the human proteins;
(ii) excluding membrane proteins and antibody proteins from the selected proteins in step (i);
(iii) selecting proteins having 10 or fewer known interactions from the rest of the proteins in step (ii); and
(iv) selecting proteins capable of forming any one or more of monomers, homodimers, and homotrimers from the selected proteins in step (iii).

5. The method according to claim 1 , wherein the selection in step (a) is performed by docking simulation.

6. The method according to claim 5 , wherein the docking simulation of step (a) is a step of selecting non-antibody proteins that bind with a predetermined number or more of target amino acid residues of EGFR domain 2.

7. The method according to claim 1 , wherein in step (a), the number of the target residues is 28, and the predetermined number is 10.

Assignees

• Choi Yoon Sup
• Chung Jun Ho
• Yoo Ji Ho
• Cho Hyun Soo
• Yoon Su Min
• Kim Kyung Lock
• Ryu Sung Ho
• Kim Sang Uk
• Postech Acad Ind Found

Inventors

• Choi Yoon Sup
• Chung Jun Ho
• Yoo Ji Ho
• Cho Hyun Soo
• Yoon Su Min
• Kim Kyung Lock
• Ryu Sung Ho
• Kim Sang Uk

Classifications

• G16B35/00

  ICT specially adapted for in silico combinatorial libraries of nucleic acids, proteins or peptides · CPC title

• C07K14/485 Primary

  Epidermal growth factor [EGF], i.e. urogastrone · CPC title

• G16B15/00

  ICT specially adapted for analysing two-dimensional [2D] or three-dimensional [3D] molecular structures, e.g. structural or functional relations or structure alignment · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title

• G16C20/60

  In silico combinatorial chemistry · CPC title