Process for preparing stable antibody maytansinoid conjugates
AssigneeImmunoGen, Inc.
InventorsXinfang Li, Jared M. Worful
Life of this patent
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Abstract
The invention provides processes for manufacturing cell-binding agent-cytotoxic agent conjugates of improved stability comprising performing the modification reaction at a high pH. The inventive processes comprise contacting a cell-binding agent with a bifunctional crosslinking reagent in a solution having a pH of 7.1 to 9 to covalently attach a linker to the cell-binding agent and thereby prepare a mixture comprising cell-binding agents having linkers bound thereto.
Claims76
A process for preparing a cell-binding agent having a linker bound thereto, which process comprises contacting a cell-binding agent with a bifunctional crosslinking reagent in a solution having a pH of 7.5 to 9 to covalently attach a linker to the cell-binding agent and thereby prepare a mixture comprising cell-binding agents having linkers bound thereto.
A process for preparing a cell-binding agent-cytotoxic agent conjugate comprising a cell-binding agent chemically coupled to a cytotoxic agent through a linker, which process comprises:
- (a) contacting a cell-binding agent with a bifunctional crosslinking reagent in a solution having a pH of 7.5 to 9 to covalently attach a linker to the cell-binding agent and thereby prepare a first mixture comprising cell-binding agents having linkers bound thereto,
- (b) subjecting the first mixture to tangential flow filtration, selective precipitation, non-adsorptive chromatography, adsorptive filtration, adsorptive chromatography, or a combination thereof and thereby prepare a purified first mixture of cell-binding agents having linkers bound thereto,
- (c) conjugating a cytotoxic agent to the cell-binding agents having linkers bound thereto in the purified first mixture by reacting the cell-binding agents having linkers bound thereto with a cytotoxic agent in a solution having a pH of about 4 to about 9 to prepare a second mixture comprising (i) cell-binding agent chemically coupled through the linker to the cytotoxic agent, (ii) free cytotoxic agent, and (iii) reaction by-products, and
- (d) subjecting the second mixture to tangential flow filtration, selective precipitation, non-adsorptive chromatography, adsorptive filtration, adsorptive chromatography, or a combination thereof to purify the cell-binding agent chemically coupled through the linker to the cytotoxic agent from the other components of the second mixture and thereby prepare a purified second mixture of cell-binding agent chemically coupled through the linker to the cytotoxic agent.
A process for preparing a cell-binding agent-cytotoxic agent conjugate comprising a cell-binding agent chemically coupled to a cytotoxic agent through a linker, which process comprises:
- (a) contacting a cell-binding agent with a bifunctional crosslinking reagent in a solution having a pH of 7.5 to 9 to covalently attach a linker to the cell-binding agent and thereby prepare a first mixture comprising cell-binding agents having linkers bound thereto,
- (b) conjugating a cytotoxic agent to the cell-binding agents having linkers bound thereto in the first mixture by reacting the cell-binding agents having linkers bound thereto with a cytotoxic agent in a solution having a pH of about 4 to about 9 to prepare a second mixture comprising (i) cell-binding agent chemically coupled through the linker to the cytotoxic agent, (ii) free cytotoxic agent, and (iii) reaction by-products, and
- (c) subjecting the second mixture to tangential flow filtration, selective precipitation, non-adsorptive chromatography, adsorptive filtration, adsorptive chromatography, or a combination thereof, to purify the cell binding agent chemically coupled through the linker to the cytotoxic agent from the other components of the second mixture and thereby prepare a purified second mixture of cell binding agent chemically coupled through the linker to the cytotoxic agent.
A process for preparing a cell-binding agent-cytotoxic agent conjugate comprising a cell-binding agent chemically coupled to a cytotoxic agent through a linker, which process comprises:
- (a) contacting a cell-binding agent with a cytotoxic agent-linker compound comprising a cytotoxic agent chemically coupled to a linker in a solution having a pH of 7.5 to 9 to covalently attach the cytotoxic agent-linker compound to the cell-binding agent and thereby prepare a mixture comprising the cell-binding agent-cytotoxic agent conjugate
- (b) subjecting the mixture comprising the cell-binding agent-cytotoxic agent conjugate to tangential flow filtration, selective precipitation, non-adsorptive chromatography, adsorptive filtration, adsorptive chromatography or a combination thereof to purify the conjugate.
Dependencies are inferred from the claim text.