Cytotoxic CD44 antibody immunoconjugates

a technology of cytotoxic cd44 and antibody, which is applied in the field of new conjugates of antibodies with cytotoxic compounds, can solve the problems of difficult to achieve, inefficient internalization of antigens, and limited success of many of these approaches

Inactive Publication Date: 2003-06-05
BOEHRINGER INGELHEIM INT GMBH
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Problems solved by technology

Many of these approaches have met limited success, and several reasons have been discussed in the literature to explain the failure.
These concentrations are thought to be difficult to achieve with many antibody-drug conjugates because of (a) insufficient potency of many common anticancer drugs, (b) low cell surface concentrat...

Method used

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  • Cytotoxic CD44 antibody immunoconjugates
  • Cytotoxic CD44 antibody immunoconjugates
  • Cytotoxic CD44 antibody immunoconjugates

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[0076] 1. Material and Methods

[0077] 1.1. In vitro Cell Proliferation Assay

[0078] For determination of viable cells the Cell Titer 96.RTM. AQ.sub.ueous non-radioactive cell proliferation assay (Promega, Wisconsin, Wis.) was used. Five thousand cells per well were seeded into 96-well plates in 90 .mu.l medium without phenol red. Cells were allowed to settle for 1 to 3 h and then serial dilutions of the immunoconjugate in 10 .mu.l PBS were added. Cells without immunoconjugate served as negative control. Cells were incubated for 4 days at 37.degree. C. in a humified 5% CO.sub.2 atmosphere and then 20 .mu.l MTS / PMS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl- )-2H-tetrazolium / phenazine formazan) were added according to the manufacturer's recommendation. After an additional 1 to 4 h incubation at 37.degree. C., the absorbance at 490 nm was recorded using an ELISA plate reader. For each cell line, triplicates were analyzed. The percentage of the surviving cel...

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Abstract

The invention relates to novel conjugates of CD44 antibodies with cytotoxic compounds, pharmaceutical compositions comprising such compounds, and their use in tumor therapy.

Description

RELATED APPLICATIONS[0001] The present application claims priority benefit of U.S. provisional application Serial No. 60 / 307,451, filed Jul. 24, 2001.BACKGROUND[0002] The invention relates to novel conjugates of antibodies with cytotoxic compounds, pharmaceutical compositions comprising such compounds, and their use in tumor therapy.[0003] There have been numerous attempts to improve the efficacy of antineoplastic drugs by conjugating such drugs to antibodies against tumor-associated antigens in order to elevate local concentration of the drug by targeted delivery to the tumor. Many of these approaches have met limited success, and several reasons have been discussed in the literature to explain the failure. For anticancer drugs acting stoichometrically, like e.g. doxorubicin or methotrexate, relatively high intracellular concentrations are necessary to exert the required cytotoxicity. These concentrations are thought to be difficult to achieve with many antibody-drug conjugates bec...

Claims

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Application Information

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IPC IPC(8): A61K47/48
CPCA61K47/48384A61K47/48561
Inventor ADOLF, GUENTHERPATZELT, ERIKSPROLL, MARLIESHEIDER, KARL-HEINZ
Owner BOEHRINGER INGELHEIM INT GMBH
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