Potentiation of anti-CD38-immunotoxin cytotoxicity

A technique of immunotoxin, toxin, applied in the fields of immunology and tumor biology

Inactive Publication Date: 2005-03-09
RES DEVMENT FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] A deficiency of the prior art is the lack of a method to induce the expression of target molecules for immunotherapy of tumors and other disease-causing cells

Method used

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  • Potentiation of anti-CD38-immunotoxin cytotoxicity
  • Potentiation of anti-CD38-immunotoxin cytotoxicity
  • Potentiation of anti-CD38-immunotoxin cytotoxicity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] CD38 expression in normal tissues is largely restricted to the thymus

[0031] The tissue specificity of CD38 was detected by dot blot hybridization of radiolabeled CD38 nucleic acid probe to commercial (CLONTECH) tissue specific mRNA. figure 1 Hybridization results are shown. CD38 was observed to be predominantly expressed in the thymus, with significantly lower levels in the prostate.

Embodiment 2

[0033] Retinoic acid (RA) enhances the cytotoxic effect of immunotoxins by increasing the expression of CD38.

[0034] HL-60 cells were incubated with immunotoxin alone or in the presence of 5 nM retinoic acid (RA). Increasing concentrations of unconjugated IB4 monoclonal antibody were added to cells incubated with immunotoxin and retinoic acid. After 3 days, cell viability was measured by MTS assay. Briefly, a 6.5 mg / ml solution of MTS [(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-( 4-thiophenyl)-2H-tetrazolium] and 0.5mM PMS (phenazine methanesulfonate) solution.The mixed MTS / PMS solution of 20 microliters is placed in the sample containing the cells to be tested in the 96-well plate In each well, incubate the plate for 1-4 hours at 37°C in a 5% CO2 atmosphere, and then measure the amount of formazan produced by MTS cell reduction of live cells by reading the absorbance at 490 nm. figure 2 The result is shown.

[0035] Immunotoxin alone had minimal effec...

Embodiment 3

[0037] Pretreatment with all-trans retinoic acid (RA) enhanced the induced killing of HL-60 cells.

[0038] HL-60 cells were preincubated overnight in the presence or absence of 5 nM all-trans retinoic acid. Cells were washed twice and incubated in increasing concentrations of immunotoxin in the presence or absence of IB4 unconjugated anti-CD38 monoclonal antibody. After 3 days, cell viability was tested. image 3 The result is shown.

[0039] Preincubation with all-trans retinoic acid followed by immunotoxin treatment resulted in more cell death than immunotoxin treatment alone. A 100-fold excess of unconjugated anti-CD38 mAb IB4 blocked the toxicity of the immunotoxin in both cases by competing with the immunotoxin and binding of the CD38 marker on cells. These results suggest that all-trans retinoic acid (RA) causes changes in cells that make them more susceptible to immunotoxins, rather than playing a direct role in target cell death.

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Abstract

The present invention is directed to the use of agents that induce high levels of cell surface molecules to provide targets for immunotoxins directed against the same cell surface molecules. A specific example is given in which all-trans-retinoic acid (RA) is used to induce high levels of CD38 cell surface antigen expression in several myeloid and lymphoid leukemia cells. CD38 was then used as target for delivering plant toxin (gelonin) to leukemia cells. Treatment of leukemia cells with RA induced high levels of CD38 in those cells that otherwise had low CD38 expression. The RA-induced leukemia cells then became exquisitely sensitive to an immunotoxin constructed from an anti-CD38 monoclonal antibody conjugated to the plant toxin gelonin.

Description

Background of the invention [0001] field of invention [0002] The present invention relates generally to immunology and tumor biology. More specifically, the present invention relates to enhancing the cytotoxicity of anti-CD38-based immunotoxins by treating target tumor cells with drugs to increase the expression of CD38 protein therein. [0003] Description of related fields [0004] The unique molecular architecture of monoclonal antibodies to deliver drugs or toxins to the surface of undesirable tumor cells presents an attractive and potentially useful strategy. In theory, the main advantage of this targeted approach to cancer therapy lies in the selective elimination of tumor cells while reducing the toxicity of therapeutic drugs to normal non-target tissues. However, there are actually many issues that need to be addressed before immunotoxin or antibody drug treatments will actually be effective in the body. [0005] One possible limitation to the success of any t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/192A61K31/203A61K45/00A61K38/00A61K38/17A61K38/19A61K39/395A61K39/44A61K47/48A61P35/00A61P35/02A61P43/00
CPCA61K39/395A61K47/48561A61K47/48446A61K47/6819A61K47/6849A61P35/00A61P35/02A61P43/00A61K38/217A61K38/212A61K38/2013A61K38/191A61K38/19A61K31/20A61K2300/00A61K38/21
Inventor M·G·罗森布拉姆K·梅达
Owner RES DEVMENT FOUND
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