Development of Site-Specific PEGylated Granulocyte Colony Stimulating Factor With Prolonged Biological Activity
| Autor: | Sonal Datta, Girish Sahni, Monika Kumari |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Histology lcsh:Biotechnology Biomedical Engineering Bioengineering 02 engineering and technology Pharmacology Neutropenia G-CSF cancer chemotherapy 03 medical and health sciences lcsh:TP248.13-248.65 PEG ratio medicine Leukocyte proliferation neutropenia PEGylated G-CSF Original Research chemistry.chemical_classification Chemistry site-specific PEGylation Bioengineering and Biotechnology Biological activity 021001 nanoscience & nanotechnology medicine.disease prolonged biological activity of G-CSF Granulocyte colony-stimulating factor Amino acid 030104 developmental biology PEGylation 0210 nano-technology Biotechnology Cysteine |
| Zdroj: | Frontiers in Bioengineering and Biotechnology Frontiers in Bioengineering and Biotechnology, Vol 8 (2020) |
| ISSN: | 2296-4185 |
| Popis: | Currently, amino-terminal PEGylated human granulocyte colony stimulating factor (huG-CSF) is used to prevent and treat neutropenia. Although huG-CSF has been used as a drug for more than 20 years, it has three significant drawbacks: (i) it relies on PEG aldehyde for PEGylation of the alpha-amino group of the first amino acid, and this leads to non-specific PEGylation of the epsilon amino group of lysine residues within the G-CSF; (ii) longer-acting G-CSF variants are desirable to reduce the risk of chemotherapy-associated neutropenia; and (iii) G-CSF cannot be administered on the day of chemotherapy. In an attempt to overcome the above drawbacks, we engineered cysteine variants of G-CSF to facilitate the maleimide PEG-based site-specific PEGylation that leads to a highly homogenous PEGylated product. Importantly, we have demonstrated that 20 kDa thiol-reactive PEG conjugated by maleimide chemistry to the Cys2 G-CSF variant exhibits leukocyte proliferative activity similar to that of the commercially available G-CSF conjugated with aldehyde PEG in a neutropenia mice model. Moreover, we have demonstrated that PEGylation of the cysteine variant of huG-CSF with higher molecular weight PEGs, such as 30 kDa PEG and 40 kDa PEG, leads to significantly prolonged leukocyte proliferation activity compared to the variant conjugated with 20 kDa PEG. Importantly, even a half-dose of the engineered variant conjugated with 40 kDa PEG exhibited significantly longer biological activity than the commercially available 20 kDa PEGylated huG-CSF. Finally, we have demonstrated that administration of the engineered variant conjugated with 40 kDa PEG on the day of administration of cyclophosphamide for inducing neutropenia in mice can alleviate neutropenia through leukocyte proliferation. In summary, this study provides the design of site-specific PEGylated huG-CSF variants with improved therapeutic potential. It opens the possibility of long-acting and same-day prophylactic administration of G-CSF after chemotherapy drug regimens. These results may pave the way for the development of potential G-CSF derivatives possessing longer half-lives and favorable clinical attributes. |
| Databáze: | OpenAIRE |
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