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Non-pathogenic and/or attenuated bacteria capable of inducing apoptosis in macrophages, process of manufacturing and uses thereof

a technology of attenuating bacteria and macrophages, which is applied in the field of attenuating bacteria and non-pathogenic bacteria, can solve the problems of slow blood flow, ineffective delivery of nutrients and oxygen to tumors or metastases, and low anti-tumor effect of i>salmonella /i> in humans, and achieves modest

Inactive Publication Date: 2009-09-03
AETERNA ZENTARIS GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the newly formed vessels are highly disorganised with incomplete endothelial linings and blind ends, resulting in sluggish blood flow and inefficient delivery of nutrients and oxygen to the tumor or metastases.
However, the efficacy of Salmonella as an anti-tumor agent in humans was only modest.
However, at this point no quantitative information is available about the fraction of tumor cells infected by intracellular bacteria and also the nature of the infected cells is not known.
Furthermore, most TAMs also appear to have defective production of reactive oxygen and nitrogen intermediates when compared with macrophages cultured in vitro (Siegert A et al., Immunology 1999, 98: 551-556; Murdoch C et al., Int J Cancer 2005, 117: 701-708) and are impaired in phagocytosis.
However, the treatment failed to show any therapeutic effect in the 4T1 breast cancer model.

Method used

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  • Non-pathogenic and/or attenuated bacteria capable of inducing apoptosis in macrophages, process of manufacturing and uses thereof
  • Non-pathogenic and/or attenuated bacteria capable of inducing apoptosis in macrophages, process of manufacturing and uses thereof
  • Non-pathogenic and/or attenuated bacteria capable of inducing apoptosis in macrophages, process of manufacturing and uses thereof

Examples

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example 1

Methods

[0122]Plasmids. Escherichia coli strains carrying plasmids pKD3, pKD4 (Datsenko, K. A. & Wanner, B. L. Proc Natl Acad Sci USA 2000, 97: 6640-6645), and pCP20 (Cherepanov, P. P. & Wackernagel, W. Gene 1995, 158: 9-14) were obtained from the Department of Biotechnology, University of Wuerzburg. The plasmids pKD3 and pKD4 are π dependent and carry chloramphenicol and kanamycin resistance genes, respectively, flanked by FLP recombinase recognition sites (FRT sites). The pCP20 plasmid contains a temperature sensitive replicon and the yeast FLP recombinase transcribed from the IpR promoter under the control of the I cl857 repressor (Cherepanov, P. P. & Wackernagel, W. Gene 1995, 158: 9-14).

[0123]Media, Chemicals and Other Reagents. Ampicillin-, chloramphenicol- (CmR), and kanamycin-resistant (KmR) transformants were selected on trypticase soy agar (1.2% agar) (TSA) (Difco Laboratories) containing the respective antibiotic at 100, 25, and 30 μg / ml. A total of 1 mM L-arabinose (Sigma...

example 2

Expression and Secretion of the ipaB-Gene (NC—004851) of Shigella flexneri in Gram Negative Bacteria (Escherichia coli K12)

[0158]2a) Cloning of ipaB-Gene (NC—004851) of Shigelle flexneri in Secretion Plasmid

[0159]Salmonella can like Shigella induce inflammation and apoptosis of infected macrophages through activation of caspase-1 mediated by the SipB protein, which is secreted via type III secretion systems (TTSS) (Suzuki, T. et al. J Biol Chem 2005, 280: 14042-14050; Zychlinsky, A. et al. Mol Microbiol 1994, 11: 619-627; Chen, L. M. et al., Mol Microbiol 1996, 21: 1101-1115; Hilbi, H. et al. J. Biol. Chem. 1998, 273: 32895-32900). Salmonella activate caspase-1 by SipB and induce apoptosis in TAMs at early, but not late timepoints and failed to reduce the relative amounts of TAMs. In contrast, metabolically attenuated, virulent Shigella strains, but not avirulent Shigella strains, are able to activate caspase-1 and induce apoptosis in TAMs by IpaB at all timepoints in the 4T1 and th...

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Abstract

The invention relates to an non-pathogenic and / or attenuated bacterium which is capable of inducing apoptosis in macrophages.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of EP 08101045.6 filed Jan. 29, 2008 and U.S. 61 / 024,225 filed Jan. 29, 2008, each of which is incorporated herein by reference.DESCRIPTION OF THE INVENTION[0002]1. Field of the Invention[0003]The invention relates to non-pathogenic and / or attenuated bacteria which are capable of inducing apoptosis in macrophages and a process of manufacturing thereof. These non-pathogenic and / or attenuated bacteria can be used as medicaments, in particular for the treatment of various tumors.[0004]2. Background of the Invention[0005]In 1893, William B. Coley described tumor regression in patients upon acute streptococcal infections (Coley W B, Clin Orthop Relat Res, 1991: 3-11).[0006]Since then, other bacteria have been shown to infiltrate, replicate and then preferentially accumulate in tumors (Yu Y A. et al., Nat Biotechnol 2004, 22: 313-320; Jain R K & Forbes N S, Proceedings of the National Academy of Sciences 2001...

Claims

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Application Information

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IPC IPC(8): A61K45/00C12N15/74C12N1/20A61P35/00A61K35/74
CPCA61K35/74C12R1/42C12N1/20A61P29/00A61P3/10A61P31/00A61P31/04A61P31/12A61P31/16A61P35/00A61P35/02A61P37/00A61P37/02A61P37/06A61P43/00Y02A50/30C12R2001/42C12N1/205A61K35/12C07K1/22C07K14/52C12N1/00C12N1/02C12N5/0645C12Q1/00C12N2710/00061
Inventor FENSTERLE, JOACHIMGALMBACHER, KATHARINARAPP, ULFGOEBEL, WERNERHOTZ, CHRISTIAN
Owner AETERNA ZENTARIS GMBH
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