Title:PEGylation Strategy for Improving the Pharmacokinetic and Antitumoral
Activity of the IL-2 No-alpha Mutein
Volume: 29
Issue: 44
Author(s): Marianniz Díaz-Hernández*, Janoi Chang-Calderón, Miguel Angel Álvarez, Ingrid Ruiz Ramírez, Olga Lidea Fernández Saez, Armando López Medinilla, Carlos Yordan González Castillo, Claudia Diaz Borges, Sum Lai Lozada Chang, Kalet León and Tania Carmenate*
Affiliation:
- Department of Immune-regulation, Center of Molecular Immunology (Cuba), Havana, Cuba
- Department of Immune-regulation, Center of Molecular Immunology (Cuba), Havana, Cuba
Keywords:
PEGylation, Interleukin-2 (IL-2) mutant, pharmacokinetic, protein structure, antitumor activity, lower toxicity.
Abstract:
Background: In a previous work, an IL-2Rβγ biased mutant derived from human IL-2 and called
IL-2noα, was designed and developed. Greater antitumor effects and lower toxicity were observed compared
to native IL-2. Nevertheless, mutein has some disadvantages, such as a very short half-life of about 9-12 min,
propensity for aggregation, and solubility problems.
Objective: In this study, PEGylation was employed to improve the pharmacokinetic and antitumoral properties
of the novel protein.
Methods: Pegylated IL-2noα was characterized by polyacrylamide gel electrophoresis, size exclusion chromatography,
in vitro cell proliferation and in vivo cell expansion bioassays, and pharmacokinetic and antitumor
studies.
Results: IL-2noα-conjugates with polyethylene glycol (PEG) of 1.2 kDa, 20 kDa, and 40 kDa were obtained
by classical acylation. No significant changes in the secondary and tertiary structures of the modified protein
were detected. A decrease in biological activity in vitro and a significant improvement in half-life were
observed, especially for IL-2noα-PEG20K. PEGylation of IL-2noα with PEG20K did not affect the capacity
of the mutant to induce preferential expansion of T effector cells over Treg cells. This pegylated IL-2noα
exhibited a higher antimetastatic effect compared to unmodified IL-2noα in the B16F0 experimental metastases
model, even when administered at lower doses and less frequently.
Conclusion: PEG20K was selected as the best modification strategy, to improve the blood circulation time of
the IL-2noα with a superior antimetastatic effect achieved with lower doses.
