Abstract
In Brazil, there is a growing demand for specialised pharmaceuticals, and the high cost of their importation results in increasing costs, reaching US$ 1.34 billion in 2012 and US$ 1.61 billion in 2013. Worldwide expenses related to drugs could reach US$ 1.3 trillion in 2018, especially due to new treatments for hepatitis C and cancer. Specialised or high-cost pharmaceutical drugs used for the treatment of viral hepatitis, multiple sclerosis, HIV and diabetes are distributed free of charge by the Brazilian government. The glucagon peptide was included in this group of high-cost biopharmaceuticals in 2008. Although its main application is the treatment of hypoglycaemia in diabetic patients, it can also be used with patients in an alcoholic coma, for those patients with biliary tract pain, and as a bronchodilator. Therefore, in order to reduce biopharmaceutical production costs, the Brazilian government passed laws focusing on the development and increase of a National Pharmaceutical Industrial Centre, including the demand for the national production of glucagon. For that reason and given the importance and high cost of recombinant glucagon, the purpose of this study was to develop methods to improve production, purification and performance of the biological activity of recombinant glucagon. Glucagon was recombined into a plasmid vector containing a Glutathione S-transferase tag, and the peptide was expressed in a heterologous Escherichia coli system. After purification procedures and molecular analyses, the biological activity of this recombinant glucagon was examined using in vivo assays and showed a highly significant (p < 0.00001) and prolonged effect on glucose levels when compared with the standard glucagon. The experimental procedure described here facilitates the high level production of recombinant glucagon with an extended biological activity.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-015-0099-2) contains supplementary material, which is available to authorized users.
Highlights
The development of recombinant proteins has been extremely promising
The ion corresponding to the peptide was present in a fraction that eluted at 26 minutes of separation, and it was further detected by matrix-assisted laser desorption ionisation (MALDI)-TOF (Matrix Assisted Laser Desorption Ionisation Time-ofFlight) MS (Additional file 2: Figure S2)
In 2016, the medicines market will spend US$ 1.2 trillion, most of which are expenses focused on cancer, diabetes and asthma drugs (IMS Institute 2012)
Summary
In 2009, pharmaceutical industries spent US$ 90 billion on the activity evaluation of 400 different drugs. In 2012, more than 150 recombinant drugs were approved by FDA (Food and Drug Administration) regulators and/or by the European Medicine Agency (Huang et al 2012). In Brazil, the Government provides free, high-cost drugs to the population, which are classified into three different categories: basic, strategic and specialised. The specialised components are expensive, which limits access of the population to important medicines. In 1993, the Program for Specialized Medicaments (SM) became a prospective solution to this problem. Most of these medicines are imported from other countries, resulting in a large cost to the Brazilian government. The expectation is that the amount spent on the importation of SM will continue to grow over the several years
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