Application of SPRA Technology for Delivery of Erythropoietin: Stability Evaluation of Conjugated Erythropoietin with Adamantane and in SPRA Inclusion Complex.
Erythropoietin
Host-guest Interaction
Poly (Ethylene Glycol)
SPRA Chemistry
Stability
Supramolecular
β-Cyclodextrin
Journal
Iranian journal of pharmaceutical research : IJPR
ISSN: 1726-6890
Titre abrégé: Iran J Pharm Res
Pays: Netherlands
ID NLM: 101208407
Informations de publication
Date de publication:
Dec 2022
Dec 2022
Historique:
received:
18
12
2022
revised:
07
01
2023
accepted:
08
01
2023
entrez:
10
3
2023
pubmed:
11
3
2023
medline:
11
3
2023
Statut:
epublish
Résumé
As a widely used therapeutic protein, recombinant human erythropoietin (rhEPO) is currently one of the most effective biopharmaceuticals on the market for the treatment of anemia in patients with chronic renal disease. Increasing in vivo rhEPO half-life and its bioactivity is a significant challenge. It was hypothesized that the application of self-assembly PEGylation retaining activity, named supramolecular (SPRA) technology, could prolong the protein half-life without a significant loss of bioactivity. This study aimed to assess the stability of rhEPO during synthetic reactions, including the conjugation with adamantane and the formation of the SPRA complex. To do this, the secondary structure of the protein was also evaluated. FTIR, ATR-FTIR, Far-UV-CD, and SDS-PAGE methods were employed. Thermal stability studies of SPRA-rhEPO complex and rhEPO were investigated at 37°C for ten days using a nanodrop spectrophotometer. The secondary structure of lyophilized rhEPO, AD-rhEPO, and rhEPO (pH 8) was compared to rhEPO. Results showed that the secondary structure of the protein was unaffected by lyophilization, pH change, and the formation of covalent bonds in conjugation reaction. SPRA-rhEPO complex was also stable for seven days in phosphate buffer (pH 7.4) at 37°C. It was concluded that the stability of rhEPO could increase by complexation using SPRA technology.
Sections du résumé
Background
UNASSIGNED
As a widely used therapeutic protein, recombinant human erythropoietin (rhEPO) is currently one of the most effective biopharmaceuticals on the market for the treatment of anemia in patients with chronic renal disease. Increasing in vivo rhEPO half-life and its bioactivity is a significant challenge. It was hypothesized that the application of self-assembly PEGylation retaining activity, named supramolecular (SPRA) technology, could prolong the protein half-life without a significant loss of bioactivity.
Objectives
UNASSIGNED
This study aimed to assess the stability of rhEPO during synthetic reactions, including the conjugation with adamantane and the formation of the SPRA complex. To do this, the secondary structure of the protein was also evaluated.
Methods
UNASSIGNED
FTIR, ATR-FTIR, Far-UV-CD, and SDS-PAGE methods were employed. Thermal stability studies of SPRA-rhEPO complex and rhEPO were investigated at 37°C for ten days using a nanodrop spectrophotometer.
Results
UNASSIGNED
The secondary structure of lyophilized rhEPO, AD-rhEPO, and rhEPO (pH 8) was compared to rhEPO. Results showed that the secondary structure of the protein was unaffected by lyophilization, pH change, and the formation of covalent bonds in conjugation reaction. SPRA-rhEPO complex was also stable for seven days in phosphate buffer (pH 7.4) at 37°C.
Conclusions
UNASSIGNED
It was concluded that the stability of rhEPO could increase by complexation using SPRA technology.
Identifiants
pubmed: 36896318
doi: 10.5812/ijpr-134282
pmc: PMC9990512
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e134282Informations de copyright
Copyright © 2023, Author(s).
Déclaration de conflit d'intérêts
Conflict of Interests: The authors declared that there is no conflict of interest.
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