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An Integrated Approach for a Structural and Functional Evaluation of Biosimilars: Implications for Erythropoietin.

Gianoncelli A, Bonini SA, Bertuzzi M, Guarienti M, Vezzoli S, Kumar R, Delbarba A, Mastinu A, Sigala S, Spano P, Pani L, Pecorelli S, Memo M - BioDrugs (2015)

Bottom Line: The analogy between the originator and its biosimilar(s) is assessed through safety, purity, and potency analyses.Chemical differences were found only at the level of isoforms containing N-glycosylation; however, functional in vitro and in vivo studies did not show any significant differences from a biosimilar point of view.These rapid and economic structural and functional analyses were effective in the evaluation of the biosimilarity between the originator rhEPO alfa and the biosimilars analyzed.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Translational Medicine, University of Brescia, Viale Europa, 11, 25123, Brescia, Italy, alessandra.gianoncelli@unibs.it.

ABSTRACT

Background: Authorization to market a biosimilar product by the appropriate institutions is expected based on biosimilarity with its originator product. The analogy between the originator and its biosimilar(s) is assessed through safety, purity, and potency analyses.

Objective: In this study, we proposed a useful quality control system for rapid and economic primary screening of potential biosimilar drugs. For this purpose, chemical and functional characterization of the originator rhEPO alfa and two of its biosimilars was discussed.

Methods: Qualitative and quantitative analyses of the originator rhEPO alfa and its biosimilars were performed using reversed-phase high-performance liquid chromatography (RP-HPLC). The identification of proteins and the separation of isoforms were studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and two-dimensional gel electrophoresis (2D-PAGE), respectively. Furthermore, the biological activity of these drugs was measured both in vitro, evaluating the TF-1 cell proliferation rate, and in vivo, using the innovative experimental animal model of the zebrafish embryos.

Results: Chemical analyses showed that the quantitative concentrations of rhEPO alfa were in agreement with the labeled claims by the corresponding manufacturers. The qualitative analyses performed demonstrated that the three drugs were pure and that they had the same amino acid sequence. Chemical differences were found only at the level of isoforms containing N-glycosylation; however, functional in vitro and in vivo studies did not show any significant differences from a biosimilar point of view.

Conclusion: These rapid and economic structural and functional analyses were effective in the evaluation of the biosimilarity between the originator rhEPO alfa and the biosimilars analyzed.

No MeSH data available.


Related in: MedlinePlus

Sigmoidal dose response curve of three different rhEPOs; EPREX® (black), Binocrit® (blue), and Retacrit® (green). BIN Binocrit®, EPO rhEPO, EPR Eprex®, RET Retacrit®
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Fig7: Sigmoidal dose response curve of three different rhEPOs; EPREX® (black), Binocrit® (blue), and Retacrit® (green). BIN Binocrit®, EPO rhEPO, EPR Eprex®, RET Retacrit®

Mentions: As reported in Fig. 7, exposure of TF-1 cells to increasing concentrations of EPR, BIN, or RET induced a stimulation of the cell proliferation rate. Comparison of the concentration–response curves between drugs did not show any significant differences (p = 0.6748; F = 0.3941); indeed, the EC50 values were 0.22 (95 % CI 0.09462–0.5304), 0.19 (95 % CI 0.1069–0.3355), and 0.30 IU/mL (95 % CI 0.1592–0.5781) for EPR, BIN, and RET, respectively. Time-course experiments up to 120 h, at the respective EC50 values, demonstrated that the reduction of the viability reached its maximum at 72 h and did not change afterwards (data not shown).Fig. 7


An Integrated Approach for a Structural and Functional Evaluation of Biosimilars: Implications for Erythropoietin.

Gianoncelli A, Bonini SA, Bertuzzi M, Guarienti M, Vezzoli S, Kumar R, Delbarba A, Mastinu A, Sigala S, Spano P, Pani L, Pecorelli S, Memo M - BioDrugs (2015)

Sigmoidal dose response curve of three different rhEPOs; EPREX® (black), Binocrit® (blue), and Retacrit® (green). BIN Binocrit®, EPO rhEPO, EPR Eprex®, RET Retacrit®
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4562010&req=5

Fig7: Sigmoidal dose response curve of three different rhEPOs; EPREX® (black), Binocrit® (blue), and Retacrit® (green). BIN Binocrit®, EPO rhEPO, EPR Eprex®, RET Retacrit®
Mentions: As reported in Fig. 7, exposure of TF-1 cells to increasing concentrations of EPR, BIN, or RET induced a stimulation of the cell proliferation rate. Comparison of the concentration–response curves between drugs did not show any significant differences (p = 0.6748; F = 0.3941); indeed, the EC50 values were 0.22 (95 % CI 0.09462–0.5304), 0.19 (95 % CI 0.1069–0.3355), and 0.30 IU/mL (95 % CI 0.1592–0.5781) for EPR, BIN, and RET, respectively. Time-course experiments up to 120 h, at the respective EC50 values, demonstrated that the reduction of the viability reached its maximum at 72 h and did not change afterwards (data not shown).Fig. 7

Bottom Line: The analogy between the originator and its biosimilar(s) is assessed through safety, purity, and potency analyses.Chemical differences were found only at the level of isoforms containing N-glycosylation; however, functional in vitro and in vivo studies did not show any significant differences from a biosimilar point of view.These rapid and economic structural and functional analyses were effective in the evaluation of the biosimilarity between the originator rhEPO alfa and the biosimilars analyzed.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Translational Medicine, University of Brescia, Viale Europa, 11, 25123, Brescia, Italy, alessandra.gianoncelli@unibs.it.

ABSTRACT

Background: Authorization to market a biosimilar product by the appropriate institutions is expected based on biosimilarity with its originator product. The analogy between the originator and its biosimilar(s) is assessed through safety, purity, and potency analyses.

Objective: In this study, we proposed a useful quality control system for rapid and economic primary screening of potential biosimilar drugs. For this purpose, chemical and functional characterization of the originator rhEPO alfa and two of its biosimilars was discussed.

Methods: Qualitative and quantitative analyses of the originator rhEPO alfa and its biosimilars were performed using reversed-phase high-performance liquid chromatography (RP-HPLC). The identification of proteins and the separation of isoforms were studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and two-dimensional gel electrophoresis (2D-PAGE), respectively. Furthermore, the biological activity of these drugs was measured both in vitro, evaluating the TF-1 cell proliferation rate, and in vivo, using the innovative experimental animal model of the zebrafish embryos.

Results: Chemical analyses showed that the quantitative concentrations of rhEPO alfa were in agreement with the labeled claims by the corresponding manufacturers. The qualitative analyses performed demonstrated that the three drugs were pure and that they had the same amino acid sequence. Chemical differences were found only at the level of isoforms containing N-glycosylation; however, functional in vitro and in vivo studies did not show any significant differences from a biosimilar point of view.

Conclusion: These rapid and economic structural and functional analyses were effective in the evaluation of the biosimilarity between the originator rhEPO alfa and the biosimilars analyzed.

No MeSH data available.


Related in: MedlinePlus