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An anti-cancer cytotoxic monoclonal antibody

A monoclonal antibody, cytotoxic technology, applied in the direction of antibodies, fusion cells, radioactive carriers, etc., can solve the problem of insufficient cancer

Inactive Publication Date: 2010-06-16
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, the assumption that drug molecules that bind to a receptor with the greatest affinity usually have the greatest likelihood of initiating or inhibiting a signal may not always be the case
[0018] Despite some advances in the treatment of breast and colon cancer, the identification and development of effective antibody therapies as single agents or co-treatments is insufficient for all types of cancer

Method used

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  • An anti-cancer cytotoxic monoclonal antibody
  • An anti-cancer cytotoxic monoclonal antibody
  • An anti-cancer cytotoxic monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Hybridoma production--hybridoma cell line AR92A271.7

[0100] According to the Budapest Treaty, the hybridoma cell line AR92A271.7 was deposited in the International Depository of Canada (the International Depository Authority of Canada, IDAC) on May 29, 2007, and the Bureau of Microbiology, Health Canada (Bureau of Microbiology, HealthCanada) (Canada, Ma 1015 Arlington Street, Winnipeg, Nitoba, R3E 3R2), accession number 290507-04. Pursuant to 37 CFR 1.808, the depositor warrants that, at the time of grant of the patent, all constraints imposed on the public availability of the deposited material are irrevocable. If the depositary cannot release a viable sample, replace the deposit.

[0101] To generate hybridomas producing anticancer antibody AR92A271.7, single cell suspensions of frozen human lung adenocarcinoma tumor tissue (Genomics Collaborative, Cambridge, MA) were prepared in PBS. Prepare IMMUNEASY by mixing gently TM (Qiagen, Venlo, The Netherlands) adjuvant...

Embodiment 2

[0108] in vitro binding

[0109] AR92A271.7 monoclonal antibody was produced by growing hybridomas in CL-1000 flasks (BD Biosciences, Oakville, ON), harvested and reseeded twice / week. Standard antibody purification steps were followed using Protein G Sepharose 4 Fast Flow (Amersham Biosciences, Baie d'Urfe, QC). It is within the scope of the invention to use humanized, deimmunized, chimeric or murine monoclonal antibodies.

[0110] The relationship between AR92A271.7 and lung (A549, NCI-H23, NCI-H322M, NCI-H460 and NCI-H520), colon (Lovo), breast (MDA-MB-231), pancreas ( Combination of BxPC-3), prostate (PC-3) and ovarian (OVCAR-3) cancer cell lines, and non-cancer cell lines from skin (CCD-27sk) and lung (Hs888.Lu). All cell lines were obtained from the American Type Tissue Collection (ATCC, Manassas, VA).

[0111] By initially using DPBS (Ca-free ++ and Mg ++ ) to wash the cell monolayer and prepare the cells for FACS. Cells were then removed from their cell culture pl...

Embodiment 3

[0114] In vivo tumor experiments using A549 cells

[0115] Example 1 demonstrates that AR92A271.7 has anticancer properties against human lung cancer cell lines. To demonstrate in vivo efficacy against human lung cancer cell lines, AR92A271.7 was tested in the A549 lung cancer xenograft model. refer to Figure 4 and 5 , 6-8 week old female SCID mice were implanted with 1 million human lung cancer cells (A549) in 100 microliters of PBS solution by subcutaneous injection at the right loin. The mice were randomly divided into two treatment groups, 10 in each group. On the day after implantation, in a solution containing 2.7mM KCl, 1mM KH 2 PO 4 , 137mM NaCl and 20mM NaCl 2 HPO 4 After dilution of the diluent from the stock concentrate, each group was administered intraperitoneally with 20 mg / kg AR92A271.7 detection antibody or buffer control in a volume of 300 microliters. The antibodies and control samples were then administered weekly for the duration of the study. Tum...

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Abstract

Antibody mediated killing of cancer cells is an effective approach to treat cancer. Antibodies generated in mice, upon immunization with lung adenocarcinoma cells, are screened for cytotoxicity against a variety of cancer cell lines as the endpomt. An anti-cancer cytotoxic monoclonal antibody is isolated, produced by the hybpdoma AR92A271.7 deposited with IDAC as Accession Number 290507-04, which is cytotoxic to a lung cancer cell line and reduces tumor burden in an animal model of human lung cancer. The monoclonal antibody also binds to several cancer cell lines. There is low, but detectable, binding of the monoclonal antibody to a non-cancer cell line, however, cytotoxicity is not induced in this non-cancer cell line. This monoclonal antibody can be used to aid staging and diagnosis of cancer, and in the treatment of primary tumors and tumor metastases. This cytotoxic monoclonal antibody can also be used to deliver toxins, enzymes, radioactive compounds and hematogenous cells to cancer cells, to further aid reduction of tumor burden.

Description

[0001] Statement of Collaborative Research Agreement [0002] The present invention, as defined by the claims herein, is made within the scope of the parties involved in the joint research agreement (the "Agreement") between Arius Research Inc. and Takeda Pharmaceutical Company Limited produced as a result of activities performed. This Agreement was in effect prior to the date of this invention. field of invention [0003] The present invention relates to the isolation and production of cancerous disease-modifying antibodies (CDMABs) and to the use of these CDMABs, optionally in combination with one or more chemotherapeutic agents, in therapeutic and diagnostic procedures. The invention also relates to binding assay methods using the CDMABs of the invention. Background of the invention [0004] Monoclonal Antibodies as Cancer Therapy: Every individual with cancer is unique and different from other cancers, as is an individual's identity. Nonetheless, current treatments tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/30A61K39/395A61K47/48A61K51/10A61P35/00A61P37/04C07K16/18C07K16/28C07K16/46C07K19/00C12N5/16C12P21/08G01N33/53G01N33/574G01N33/577
CPCA61K47/48592C07K16/3023A61K2039/505A61K51/1054G01N33/57484A61K47/6857A61P35/00A61P37/04
Inventor 戴维·S·F·扬海伦·P·芬德利苏珊·E·哈恩
Owner F HOFFMANN LA ROCHE & CO AG
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