Targeting intracellular target-binding determinants with intracellular antibodies
AssigneeThe United States Government as represented by the Department of Veterans Affairs
InventorsRichard H. Weisbart, Robert N. Nishimura
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Abstract
The invention provides a method for inhibiting an intracellular target in a cell with a bispecific antibody comprising contacting the cell with a bispecific antibody having a first Fv fragment with a cell-penetrating determinant and a second Fv fragment with an intracellular target-binding determinant under suitable conditions so that the first Fv fragment causes the bispecific antibody to enter the cell and the second Fv fragment binds the intracellular target in the cell and thereby inhibiting the intracellular target.
Claims21
A bispecific antibody comprising a first FV fragment with a cell-penetrating determinant from an anti-DNA monoclonal antibody 3E10 or an antibody which competes with monoclonal antibody 3E10 and a second Fv fragment with an intracellular target-binding determinant that increases the biological activity, biochemical activity, regulatory activity or cellular signal associated with the determinant or a macromolecule to which the determinant is attached, wherein the second Fv fragment is directed to mutant p53 protein, wherein the first Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRH1 comprising SEQ ID NO:
- 11,
- b) CDRH2 comprising SEQ ID NO:
- 13, and
- c) CDRH3 comprising SEQ ID NO:
- 15, and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRL1 comprising SEQ ID NO:
- 5,
- b) CDRL2 comprising SEQ ID NO:
- 7 and
- c) CDRL3 comprising SEQ ID NO:
- 9, and,
- wherein the second Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRH1 comprising SEQ ID NO:
- 38,
- b) CDRH2 comprising SEQ ID NO:
- 40, and
- c) CDRH3 comprising SEQ ID NO:
- 42, and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRL1 comprising SEQ ID NO:
- 32,
- b) CDRL2 comprising SEQ ID NO:
- 34 and
- c) CDRL2 comprising SEQ ID NO:
- 36.
A method for increasing p53 tumor suppressor protein levels in a tumor or cancer cell by exposing the tumor or cancer cell with a bispecific antibody having a first Fv fragment with a cell-penetrating determinant and a second Fv fragment with an intracellular target-binding determinant, wherein the second Fv fragment is directed to a mutant p53 protein, and wherein the bispecific antibody binds to the mutant p53 protein, thereby increasing the level of p53 tumor suppressor protein levels in the tumor or cancer cell,
- wherein the first Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- d) CDRH1 comprising SEQ ID NO:
- 11,
- e) CDRH2 comprising SEQ ID NO:
- 13, and
- f) CDRH3 comprising SEQ ID NO:
- 15, and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- d) CDRL1 comprising SEQ ID NO:
- 5,
- e) CDRL2 comprising SEQ ID NO:
- 7 and
- f) CDRL3 comprising SEQ ID NO:
- 9, and,
- wherein the second Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRH1 comprising SEQ ID NO:
- 38,
- b) CDRH2 comprising SEQ ID NO:
- 40, and
- c) CDRH3 comprising SEQ ID NO:
- 42, and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRL1 comprising SEQ ID NO:
- 32,
- b) CDRL2 comprising SEQ ID NO:
- 34 and
- c) CDRL3 comprising SEQ ID NO:
- 36.
A method for inhibiting the growth of tumor or cancer cells associated with p53 expression or activity in a subject by exposing the tumor or cancer cells to a bispecific antibody having a first Fv fragment with a cell-penetrating determinant of a 3E10 monoclonal antibody and a second Fv fragment with an intracellular target-binding determinant that inhibits an intracellular target under suitable conditions so that the first Fv fragment causes the bispecific antibody to enter the cells and the second Fv fragment binds the intracellular target in the cells and inhibits the intracellular target, wherein the first Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRH1 comprising SEQ ID NO:
- 11,
- b) CDRH2 comprising SEQ ID NO:
- 13 and
- c) CDRH3 comprising SEQ ID NO:
- 15;
- and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRL1 comprising SEQ ID NO:
- 5,
- b) CDRL2 comprising SEQ ID NO:
- 7, and
- c) CDRL3 comprising SEQ ID NO:
- 9;
- and
- wherein the second Fv fragment comprises a heavy chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRH1 comprising SEQ ID NO:
- 38,
- b) CDRH2 comprising SEQ ID NO:
- 40 and
- c) CDRH3 comprising SEQ ID NO:
- 42;
- and,
- a light chain variable domain comprising the following complementarity determining region (CDR) amino acid sequences:
- a) CDRL1 comprising SEQ ID NO:
- 32,
- b) CDRL2 comprising SEQ ID NO:
- 34, and
- c) CDRL3 comprising SEQ ID NO:
- 36,
- thereby inhibiting the growth of tumor or cancer cells that express mutant p53 protein in the subject.
Dependencies are inferred from the claim text.