Kinase mutants and uses thereof

Patent metadata

Field	Value
Publication number	US-12478626-B2
Application number	US-202318348175-A
Country	US
Kind code	B2
Filing date	Jul 6, 2023
Priority date	May 24, 2017
Publication date	Nov 25, 2025
Grant date	Nov 25, 2025

Abstract

The present invention relates to methods of designing kinase mutants for reprogramming the sensitivity of a target kinase to some specific inhibitors, methods of reprogramming the sensitivity of a target kinase to some specific inhibitors, wherein those kinase inhibitors have little or no affinity for the wild-type target kinase, vectors or cells expressing said mutated kinases, composition and uses thereof for the prevention and/or treatment of a disease or disorder, in particular cancer.

First claim

The invention claimed is:

1 . An isolated nucleic acid molecule encoding a Zap-70 kinase mutant that retains functional activity of wild-type Zap-70 kinase and is more sensitive than wild-type Zap-70 kinase to inhibition by erlotinib and vandetanib, wherein the Zap-70 kinase mutant comprises an amino acid sequence of SEQ ID NO: 1 wherein is introduced at least one mutation selected from: V399L, M414A, M414T, M414V, M416Y, M416V, M416L, M416L, M416F, M416W, M416H, M416T, and M416S.

2 . The isolated nucleic acid molecule of claim 1 , wherein the Zap-70 kinase mutant comprises an amino acid sequence selected from SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

3 . The isolated nucleic acid molecule of claim 1 , wherein the Zap-70 kinase mutant comprises an amino acid sequence of SEQ ID NO: 1, wherein is introduced the mutation M414A, and at least one conservative substitution of at least one amino acid on at least one position selected from 342, 344-346, 350-354, 367-369, 386, 390, 399, 412, 413, 415-421, 424, 466-468, and 478-480.

4 . The isolated nucleic acid molecule of claim 1 , wherein the Zap-70 kinase mutant further comprises at least one conservative substitution of at least one amino acid of the mutated SEQ ID NO: 1 on at least one position selected from 342, 344-346, 350-354, 367-369, 386, 390, 399, 412-421, 424, 466-468, and 478-480, which was not mutated.

5 . A recombinant vector comprising the nucleic acid molecule of claim 1 .

6 . A pharmaceutical composition comprising the recombinant vector of claim 5 and at least one pharmaceutically acceptable carrier, diluent, or excipient.

7 . An isolated cell comprising the nucleic acid molecule of claim 1 .

8 . The isolated cell of claim 7 , wherein the cell is a T cell.

9 . A pharmaceutical composition comprising the cell of claim 7 and at least one pharmaceutically acceptable carrier, diluent, or excipient.

10 . An isolated nucleic acid molecule encoding a Zap-70 kinase mutant that retains functional activity of wild-type Zap-70 kinase and is more sensitive than wild-type Zap-70 kinase to inhibition by erlotinib and vandetanib, wherein the Zap-70 kinase mutant comprises an amino acid sequence selected from SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8.

11 . A recombinant vector comprising the nucleic acid molecule of claim 10 .

12 . A pharmaceutical composition comprising the recombinant vector of claim 11 and at least one pharmaceutically acceptable carrier, diluent, or excipient.

13 . An isolated cell comprising the nucleic acid molecule of claim 10 .

14 . The isolated cell of claim 13 , wherein the cell is a T cell.

15 . A pharmaceutical composition comprising the cell of claim 13 and at least one pharmaceutically acceptable carrier, diluent, or excipient.

Assignees

• Sib Swiss Inst Of Bioinformatics
• Chuv—Centre Hospitalier Univ Vaudois
• Univ Lausanne
• Ludwig Inst For Cancer Res Ltd
• Chuv Centre Hospitalier Univ Vaudois

Inventors

• Michielin Olivier
• Zoete Vincent
• Coukos George
• Irving Melita
• Dilek Nahzli
• Reichenbach Patrick
• Ascencao Kelly

Classifications

• G01N33/573

  for enzymes or isoenzymes · CPC title

• C12Y207/10

  Protein-tyrosine kinases (2.7.10) · CPC title

• C12Q1/485 Primary

  involving kinase · CPC title

• C12N15/1024

  In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair · CPC title

• C12N9/1205

  Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases · CPC title