Development of a new PEG conjugated asparaginase for treatment of neoplasias

• Grant number: 19/12401-6
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: September 01, 2020
• End date: August 31, 2022
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Carlos Alexandre Breyer
• Grantee: Carlos Alexandre Breyer
• Company: Biobreyer Pesquisa e Desenvolvimento Científico Ltda
• CNAE: Pesquisa e desenvolvimento experimental em ciências físicas e naturais ; Atividades profissionais, científicas e técnicas não especificadas anteriormente
• Principal investigators: Renata Bannitz Fernandes
• Associated research grant: 17/08591-9 - Development of a new PEG conjugated asparaginase for the neoplasias treatment, AP.PIPE
• Associated scholarship(s): 21/13411-5 - Development of a new PEG conjugated Asparaginase for treatment of neoplasias, BP.PIPE; 20/14051-0 - Development of a new Asparaginase conjugated with PEG for the treatment of neoplasms: technical training in management and innovation, BP.TT; 20/12097-2 - Development of a new PEG conjugated Asparaginase for treatment of neoplasias, BP.PIPE

Abstract

Asparaginase (ASNase) is an enzyme used to treat lymphatic system cancers, especially Acute Lymphoblastic Leukemia (ALL). The ASNase administration can promote adverse effects in the patients, including immunological responses, allergic reactions, and anaphylactic shock. In these cases, treatment with other ASNase formulations is possible, however there are still adverse responses in many patients, and most countries do not offer all formulations for treatment because of the high cost and the lack of registration in regulatory agencies. Therefore, the search for new ASNase sources and new variants of the available ASNase is focus of several studies around the world. Additionally, supply crises have been frequent in many countries, and Brazil is facing it since 2011. In this PIPE Phase 2 project, we are proposing the production process development of the engineered ASNase from Escherichia coli (EcAMUT) conjugated with polyethylene glycol (PEG), giving continuity to the results obtained in the PIPE Phase 1. This proposal have five complementary objectives: 1) E. coli strain development for heterologous EcAMUT expression (IP-free); 2) Determination of the culture conditions on a bench scale and in bioreactor (fedd-batch) in high cell density (> 50g dry mass/L); 3) Enzyme clarification and purification before and after PEG conjugation; 4) Assays using leukemic cell lines (citotocicity) and with in vivo model (antibody formation and enzyme half-life); 5) Technical-economic analysis of the production process. Therefore, this project aims to develop a viable process for PEG-EcAMUT production, providing conditions for the scale up and preclinical studies. Thus, the project will result in an asparaginase biobetter to supply the national demand and will represent a global innovation (global market), since PEG-EcAMUT has improved characteristics compared to competitors. Additionally, we believe that the PEG-EcAMUT development has great importance for the biopharmaceutical processes development in Brazil, especially because it is an injectable drug. (AU)