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Liposomal Preparations against Drug Resistant Tumors

#2288


Development and Study of Liposomal Preparations for Therapy of Drug Resistant Tumors

Tech Area / Field

  • BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
  • MED-OTH/Other/Medicine

Status
3 Approved without Funding

Registration date
20.08.2001

Leading Institute
State Research Center for Applied Microbiology, Russia, Moscow reg., Obolensk

Supporting institutes

  • Cancer Research Center / Research Institute of Experimental Diagnostics and Therapy of Tumors, Russia, Moscow\nNIIIT (Pulse Techniques), Russia, Moscow

Collaborators

  • Northeastern University / Bouve College of Health Sciences, USA, MA, Boston

Project summary

The Project goal is development of laboratory preparations for tumor chemotherapy on the basis of "stealth” liposomes, immunoliposomes, and "stealth-immunoliposomes", capable of significantly decreasing or overcoming melanoma drug resistance, and also the study of their effect in vitro.

Therefore, the Project tasks are:

  • development of anti-melanoma associated antigen immunoglobulin IgG(ICO218) derivatives, single-chain variable fragment scFv(ICO218), and its hydrophobic ‘anchor’ tagged variant, providing easy incorporation of the mini-antibody into the walls of liposomal vesicles, by using genetic engineering techniques;
  • isolation and purification of the proteins in preparative amounts;
  • development of "stealth liposomes", capable of prolonged circulation in the blood of patients by using PEGylated phospholipids;
development of ‘immunoliposomes’ carrying immunoglobulin IgG(ICO218) or its fragment on the liposome vesicle outer surface, and polymer-coated ‘stealth-immunoliposomes’;
  • development of formulations containing aranose and/or nitrullin in ‘stealth-immunoliposomes' and the study of their physico-chemical properties;
  • study of the anti-tumor effect of the liposomal preparations of aranose and nitrullin in tumor cell cultures, resistant and non-resistant to chemotherapeutic agents;
  • study of the preparation mechanisms of action, the processes of internalization, effect on Pgp-170 transport functions, on transcription and translation of Pgp170 biosynthesis in the cell;
  • study of anti-cancer activity of the preparations developed using immune deficient mice inoculated with human melanoma as an in vivo model.
The impact of the proposed project on progress in oncology, cell biology and drug development will be felt in the study of the important practical problems of the multidrug resistance (MDR) of tumors. By using a comparative study of the action of novel free and liposomal anti-tumor preparations on the molecular and cell levels, progress in complete understanding and control of complicated mechanisms of cell resistivity to stresses is expected.
Prerequisites for Project fulfillment:
1) Technology of liposomes with required mean vesicle sizes is elaborated;
2) New anti-cancer preparations, aranose and nitrullin, are developed and tested;
3) Hybridoma ICO218 producer of melanoma associated antigens is developed and melanoma cell lines with MDR are obtained;
4) Plasmid vectors, suitable for carrying gene encoding scFv(ICO218), are available;
5) Experienced personnel, facilities and equipment are available for work on the project, and vivarium, to maintain the immune deficient mice;
6) Technical resources are at hand for the development and production of non-standardized equipment needed for successful project execution.
Methods of modern molecular biology and genetic engineering will be engaged in hybridoma ICO218 immunoglobuline variable fragment cloning into E.coli and the development of modified mini-antibody genes. Biochemistry and biophysics methods will also be used, for isolation and purification of mini-antibodies, development of liposomes, immunoliposomes and ‘stealth’ liposomes. Biotechnology (works on mammalian and bacterial cell cultures), oncology (testing of the developed preparations) and cytology (study of the means of action of liposomal preparations) will also be applied.
Expected Project results:
1) microbiological strains, capable of protein scFv(ICO218) and its hydrophobized variant production;

2) laboratory preparations of «stealth-immunoliposomes»;

3) laboratory preparations of aranose and nitrullin, capable of overcoming MDR tumors in vitro;

4) protocols of trials of the anti-tumour effect of these preparations in cell cultures;

5) protocols of trials of the developed preparations in immune deficient mice in vivo.
The development of novel anti-cancer preparations is expected upon project completion. The preparations can be applied to cancer disease treatment, upon completion of the appropriate trials.
The proposed project continues works on development of anti-cancer recombinant proteins, which were started several years ago at the State Research Center for Applied Microbiology with the development of hybrid proteins TNF-thymosin, IFNg-miniAb(ICO-25), and SEA-scFv(ICO-25) within ISTC Project №118B. The project also continues works on the development and testing of novel anti-cancer preparations in the All-Russian Center of Oncology of the Russian Academy of Medicine.
Foreign collaborator, Professor V.P. Torchilin (Northeastern University, Boston, USA) is ready to participate in discussion of the experiments, exchange cell lines, genetic constructions and other biological materials. During a preliminary visit to his laboratory in August 2001, key persons of the project were introduced to his methods of investigations and to the main results in targeted drugs development. Agreement was achieved on a regular exchange of information and ideas, and the Project participants were invited to make some experiments, using Prof. Torchilin’s laboratory.


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