L-Asparaginase encapsulated intact erythrocytes for treatment of acute lymphoblastic leukemia (ALL)

Kwon, Young Min; Chung, Hee Soon; Moon, Cheol; Yockman, James W.; Park, Yoon Jeong; Gitlin, Scott D.; David, Allan E.; Yang, Victor C.

doi:10.1016/j.jconrel.2009.06.027

Cited by 110 publications

(75 citation statements)

References 33 publications

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“…Normally, human RBC have a life-span of 120 days 48 , which is significantly longer than the half-life of dmCocE (~4.5 hours) and the life-span of any known synthetic drug carrier. In our previous publication, we already confirmed the feasibility of this RBC encapsulation strategy by using the clinical protein drug asparaginase as an example 11 . Another potential application of CPP-CocE products is through nasal delivery.…”

Section: Discussionsupporting

confidence: 53%

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Cell Permeable Cocaine Esterases Constructed by Chemical Conjugation and Genetic Recombination

et al. 2012

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Cocaine esterase (CocE) is the most efficient cocaine-metabolizing enzyme tested in vivo to date, displaying a rapid clearance of cocaine and a robust protection against cocaine’s toxicity. Two potential obstacles to the clinical application of CocE, however, lie in its proteolytic degradation and induced immune response. To minimize these potential obstacles, we attempted non-disruptive cell encapsulation by creating a cell permeable form of CocE, which was achieved by covalently linking a thermally stable CocE mutant (dmCocE) with cell penetrating peptides (CPPs). Two types of CPPs, Tat and the low molecular weight protamine (LMWP), were used in this study. Two types of disulfide-bridged chemical conjugates, Tat-S-S-dmCocE and LMWP-S-S-dmCocE, were synthesized and then purified by heparin affinity chromatography. In addition, four recombinant CPP-dmCocE fusion proteins, Tat-N-dmCocE, LMWP-N-dmCocE, dmCocE-C-Tat, and dmCocE-C-LMWP, were constructed, expressed in E. coli and purified as soluble proteins. Among these six CPP-dmCocE variants, LMWP-S-S-dmCocE showed the highest cocaine-hydrolyzing activity, and dmCocE-C-Tat had the highest production yield. To evaluate their cellular uptake behavior, a covalently-linked fluorophore (FITC) was utilized to visualize the cellular uptake of all six CPP-dmCocE variants in living HeLa cells. All the six variants exhibited cellular uptake, but their intracellular distribution phenotypes differed. While the chemical conjugates showed primarily cytoplasmic distribution, which was likely due to the reduction of the disulfide linkage between CPP and dmCocE, all the other four recombinant fusion proteins displayed both nuclear and cytoplasmic localization, with dmCocE-C-CPP exhibiting higher cytoplasmic distribution during cellular uptake. Based on a balanced consideration of essentials for clinical application, including parameters such as high cocaine-hydrolyzing efficiency, large production yield, major cytoplasmic distribution, etc., the dmCocE-C-Tat fusion protein seems to be the best candidate from this investigation. Further in vivo studies of the cell-encapsulated dmCocE-C-Tat in hydrolyzing cocaine and alleviating immunogenicity and proteolytic degradation in established, clinically relevant mouse models are currently underway in our laboratories. Findings from this research are not only useful for developing other new CPP-CocE constructs, but also valuable for establishing a non-disruptive cell-encapsulation technology for other protein therapeutics that are known to be immunogenic for direct clinical application.

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Section: Discussionsupporting

confidence: 53%

“…Based on the results presented in this research, further studies that involve the applications of cell-permeable CPP-CocE products in established, clinically relevant mouse models 8, 11 are currently underway in our laboratories. One potential clinical application of CPP-CocE is CocE-encapsulated RBC.…”

Section: Discussionmentioning

confidence: 99%

Cell Permeable Cocaine Esterases Constructed by Chemical Conjugation and Genetic Recombination

et al. 2012

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“…Currently, phase II/III clinical trials, for the treatment of ALL patients with L-Asparaginase entrapped into human RBC, are ongoing in Europe. Recently, L-Asparaginase has been encapsulated into RBC by means of a novel method using the membrane-traslocating low molecular weight protamine (88). This method was demonstrated to give intact and fully functional RBC and prolong enzyme half-life, resulting very effective in the treatment of LLA in mouse models.…”

Section: Red Blood Cells As Enzyme Carriersmentioning

confidence: 99%

Drug delivery by red blood cells

et al. 2011

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SummaryDrug delivery is a growing field of interdisciplinary activities that combine the use of new materials with the biochemical properties of selected drugs, with the aim of improving their therapeutic action and reducing their toxicity. In few cases, proper medical devices have been also realized to implement new drug delivery modalities. In this article, we have summarized available information and our experience on the use of autologous Red Blood Cells as carriers for drugs to be released within the vascular system. This is not a comprehensive review, but it focusses on the mechanisms that are available to distribute drugs in circulation by carrier red blood cells and provide illustrative examples on how this is currently obtained. We have not included a summary of clinical data collected in recent years using this technology but simply provided proper references for the interested readers. Finally, a special attention is devoted to the possibility of entrapping, into autologous red blood cells, recombinant drug-binding proteins. This new strategy is opening the way at a new modality to influence the vascular distribution of drugs by realizing a dynamic circulating container (the engineered red cell) capable of reversible binding and transportation of one or more drugs of interest selected on the bases of the red cell entrapped target proteins. This new modality is not yet fully developed and explored but will certainly provide a technical solution to the problem of stabilizing drug concentration in circulation improving drug efficacy and reducing drug toxicity.

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“…Encapsulation of L-asparaginase (ASNase) into red blood cells (RBC) has been popular criteria to circumvent immunogenicity from the exogenous protein. Kwon et al (2009) developed an innovative method for encapsulation of therapeutically active ASNase into functionally intact erythrocytes and found that erythrocytes treated by this encapsulation method not only exhibited a long plasma half-life similar to that of untreated RBCs, but also displayed enhanced therapeutic effects of the entrapped protein drug owing to protection of the drug by erythrocyte from proteolytic degradation and phagocytic clearance. Advantages of erythrocytes as the drug carrier to overcome toxicity issues related to use of free ASNase was also reviewed by Hamidi et al (2007).…”

Section: Resealed Erythrocytes As Protein Stabilizermentioning

confidence: 99%