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Impact of Silicone Oil on Free Fatty Acid Particle Formation due to Polysorbate 20 Degradation

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Abstract

Purpose

Polysorbate 20 (PS20), a commonly used surfactant in biopharmaceutical formulations, can undergo hydrolytic degradation resulting in free fatty acids (FFAs) that precipitate to form particles. This work investigates the ability for silicone oil (si-oil) coated on the interior walls of prefilled syringes (PFSs) to act as a sink for FFAs and potentially delay FFA particle formation.

Methods

Myristic acid distribution coefficient was measured in a two-phase system containing si-oil and formulation buffer at a range of aqueous conditions. An empirical model was built from these data to predict distribution coefficient based on aqueous conditions. To verify the model, PS20 was degraded using model lipases side-by-side in glass vials and PFSs while monitoring sub-visible particles.

Results

The empirical model demonstrates that the partitioning of myristic acid into si-oil is maximized at low pH and low PS20 concentration. The model predicts that the presence of si-oil at levels typical in PFSs provides at most an 8.5% increase in the total carrying capacity for myristic acid compared to a non-coated glass vial. The time to onset of FFA particles was equivalent between degradations performed in two PFS models coated with differing levels of silicone oil and in non-coated glass vials.

Conclusion

Herein, we demonstrate that FFAs partition from aqueous solution into si-oil. However, the extent of the partitioning effect is not large enough to delay PS20-related FFA particle formation at typical formulation conditions (pH 5.0–7.5, 0.01% - 0.1% w/v PS20) filled in typical PFSs (<1.0 mg si-oil/mL aqueous fill).

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Abbreviations

FFA:

Free fatty acid

PFS:

Pre-filled syringe

PS20:

Polysorbate-20

SI-OIL:

Silicone oil

FB:

Formulation buffer

D:

Distribution coefficient

P:

Partition coefficient

PCL:

Pseudomonas cepacea lipase

CALB:

Candida antarctica lipase B

RP-UHPLC:

Reverse-phase ultra-high performance liquid chromatography

MM-HPLC:

Mixed-mode high performance liquid chromatography

ELSD:

Evaporative light scattering detection

RP-HPLC:

Reverse-phase high performance liquid chromatography

HA:

Protonated (non-ionized) fatty acid

A− :

Deprotonated (ionized) fatty acid

References

  1. Manning MC, Patel K, Borchardt RT. Stability of protein pharmaceuticals. Pharm Res. 1989;6(11):903–18.

    Article  CAS  PubMed  Google Scholar 

  2. Duncan MR, Lee JM, Warchol MP. Influence of surfactants upon protein/peptide adsorption to glass and polypropylene. Int J Pharm. 1995;120(2):179–88.

    Article  CAS  Google Scholar 

  3. Wang W. Instability, stabilization, and formulation of liquid protein pharmaceuticals. Int J Pharm. 1999;185(2):129–88.

    Article  CAS  PubMed  Google Scholar 

  4. Jorgensen L, Hostrup S, Moeller EH, Grohganz H. Recent trends in stabilising peptides and proteins in pharmaceutical formulation – considerations in the choice of excipients. Expert Opinion on Drug Delivery. 2009;6(11):1219–30.

    Article  CAS  PubMed  Google Scholar 

  5. Kerwin BA. Polysorbates 20 and 80 used in the formulation of protein biotherapeutics: structure and degradation pathways. J Pharm Sci. 2008;97(8):2924–35.

    Article  CAS  PubMed  Google Scholar 

  6. Ha E, Wang W, Wang YJ. Peroxide formation in polysorbate 80 and protein stability. J Pharm Sci. 2002;91(10):2252–64.

    Article  CAS  PubMed  Google Scholar 

  7. Kishore RSK, Kiese S, Fischer S, Pappenberger A, Grauschopf U, Mahler H-C. The degradation of Polysorbates 20 and 80 and its potential impact on the stability of biotherapeutics. Pharm Res. 2011;28(5):1194–210.

    Article  CAS  PubMed  Google Scholar 

  8. Labrenz SR. Ester hydrolysis of Polysorbate 80 in mAb drug product: evidence in support of the hypothesized risk after the observation of visible particulate in mAb formulations. J Pharm Sci. 2014;103(8):2268–77.

    Article  CAS  PubMed  Google Scholar 

  9. Tomlinson A, Demeule B, Lin B, Yadav S. Polysorbate 20 degradation in biopharmaceutical formulations: quantification of free fatty acids, characterization of particulates, and insights into the degradation mechanism. Mol Pharm. 2015;12(11):3805–15.

    Article  CAS  PubMed  Google Scholar 

  10. Dixit N, Salamat-Miller N, Salinas PA, Taylor KD, Basu SK. Residual host cell protein promotes Polysorbate 20 degradation in a Sulfatase drug product leading to free fatty acid particles. J Pharm Sci. 2016;105(5):1657–66.

    Article  CAS  PubMed  Google Scholar 

  11. Kishore RSK, Pappenberger A, Dauphin IB, Ross A, Buergi B, Staempfli A, et al. Degradation of polysorbates 20 and 80: studies on thermal autoxidation and hydrolysis. J Pharm Sci. 2011;100(2):721–31.

    Article  CAS  PubMed  Google Scholar 

  12. Dwivedi M, Blech M, Presser I, Garidel P. Polysorbate degradation in biotherapeutic formulations: identification and discussion of current root causes. Int J Pharm. 2018;552(1):422–36.

    Article  CAS  PubMed  Google Scholar 

  13. Doshi N, Demeule B, Yadav S. Understanding particle formation: solubility of free fatty acids as Polysorbate 20 degradation byproducts in therapeutic monoclonal antibody formulations. Mol Pharm. 2015;12(11):3792–804.

    Article  CAS  PubMed  Google Scholar 

  14. Saggu M, Liu J, Patel A. Identification of subvisible particles in biopharmaceutical formulations using Raman spectroscopy provides insight into Polysorbate 20 degradation pathway. Pharm Res. 2015;32(9):2877–88.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Chiu J, Valente KN, Levy NE, Min L, Lenhoff AM, Lee KH. Knockout of a difficult-to-remove CHO host cell protein, lipoprotein lipase, for improved polysorbate stability in monoclonal antibody formulations. Biotechnol Bioeng. 2017;114(5):1006–15.

    Article  CAS  PubMed  Google Scholar 

  16. Katz JS, Nolin A, Yezer BA, Jordan S. Dynamic properties of novel excipient suggest mechanism for improved performance in liquid stabilization of protein biologics. Mol Pharm. 2019;16(1):282–91.

    Article  CAS  PubMed  Google Scholar 

  17. Makwana S, Basu B, Makasana Y, Dharamsi A. Prefilled syringes: an innovation in parenteral packaging. Int J Pharm Investig. 2011;1(4):200–6.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Sacha G, Rogers JA, Miller RL. Pre-filled syringes: a review of the history, manufacturing and challenges. Pharm Dev Technol. 2015;20(1):1–11.

    Article  CAS  PubMed  Google Scholar 

  19. Sigma-Aldrich. Silicone Oil Safety Data Sheet (Aldrich - 378364). 2020.

  20. Doshi N, Fish R, Padilla K, Yadav S. Evaluation of super refined™ Polysorbate 20 with respect to Polysorbate degradation. Journal of Pharmaceutical Sciences: Particle Formation and Protein Stability; 2020.

    Google Scholar 

  21. Hewitt D, Zhang T, Kao YH. Quantitation of polysorbate 20 in protein solutions using mixed-mode chromatography and evaporative light scattering detection. J Chromatogr A. 2008;1215(1–2):156–60.

    Article  CAS  PubMed  Google Scholar 

  22. Hewitt D, Alvarez M, Robinson K, Ji J, Wang YJ, Kao YH, et al. Mixed-mode and reversed-phase liquid chromatography-tandem mass spectrometry methodologies to study composition and base hydrolysis of polysorbate 20 and 80. J Chromatogr A. 2011;1218(15):2138–45.

    Article  CAS  PubMed  Google Scholar 

  23. McShan AC, Kei P, Ji JA, Kim DC, Wang YJ. Hydrolysis of Polysorbate 20 and 80 by a range of Carboxylester hydrolases. PDA J Pharm Sci Technol. 2016;70(4):332–45.

    Article  CAS  PubMed  Google Scholar 

  24. Siska CC, Pierini CJ, Lau HR, Latypov RF, Fesinmeyer RM, Litowski JR. Free fatty acid particles in protein formulations, part 2: contribution of polysorbate raw material. J Pharm Sci. 2015;104(2):447–56.

    Article  CAS  PubMed  Google Scholar 

  25. Werk T, Volkin DB, Mahler HC. Effect of solution properties on the counting and sizing of subvisible particle standards as measured by light obscuration and digital imaging methods. Eur J Pharm Sci. 2014;53:95–108.

    Article  CAS  PubMed  Google Scholar 

  26. Barroso da Silva FL, Bogren D, Söderman O, Åkesson T, Jönsson B. Titration of fatty acids solubilized in cationic, nonionic, and anionic micelles. Theory and experiment. The Journal of Physical Chemistry B. 2002;106(13):3515–22.

  27. Popović MR, Popović GV, Agbaba DD. The effects of anionic, cationic, and nonionic surfactants on Acid–Base Equilibria of ACE inhibitors. J Chem Eng Data. 2013;58(9):2567–73.

    Article  CAS  Google Scholar 

  28. Tomlinson A, Zarraga IE, Demeule B. Characterization of Polysorbate Ester fractions and implications in protein drug product stability. Mol Pharm. 2020;17(7):2345–53.

    Article  CAS  PubMed  Google Scholar 

  29. Bravo B, Sánchez J, Cáceres A, Chávez G, Ysambertt F, Márquez N, et al. Partitioning of fatty acids in oil/water systems analyzed by HPLC. J Surfactant Deterg. 2008;11(1):13–9.

    Article  CAS  Google Scholar 

  30. Sangster J. Octanol-water partition coefficients of simple organic compounds. J Phys Chem Ref Data. 1989;18(3):1111–229.

    Article  CAS  Google Scholar 

  31. LOGKOW databank [Internet]. Sangster Res. Lab. 1994.

  32. Funke S, Matilainen J, Nalenz H, Bechtold-Peters K, Mahler HC, Friess W. Optimization of the bake-on siliconization of cartridges. Part I: optimization of the spray-on parameters. Eur J Pharm Biopharm. 2016;104:200–15.

    Article  CAS  PubMed  Google Scholar 

  33. Jones LS, Kaufmann A, Middaugh CR. Silicone oil induced aggregation of proteins. J Pharm Sci. 2005;94(4):918–27.

    Article  CAS  PubMed  Google Scholar 

  34. Lapelosa M, Patapoff TW, Zarraga IE. Molecular simulations of micellar aggregation of polysorbate 20 ester fractions and their interaction with N-phenyl-1-naphthylamine dye. Biophys Chem. 2016;213:17–24.

    Article  CAS  PubMed  Google Scholar 

  35. Nayem J, Zhang Z, Tomlinson A, Zarraga IE, Wagner NJ, Liu Y. Micellar morphology of Polysorbate 20 and 80 and their Ester fractions in solution via small-angle neutron scattering. J Pharm Sci. 2020;109(4):1498–508.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Purification Development, Genentech Inc, 1 DNA Way, South San Francisco, California, 94080, USA

    Raphael Fish

  2. Pharmaceutical Development, Genentech Inc., 1 DNA Way, South San Francisco, California, 94080, USA

    Jasper Lin & Nidhi Doshi

Authors
  1. Raphael Fish
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  3. Nidhi Doshi
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Correspondence to Nidhi Doshi.

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Fish, R., Lin, J. & Doshi, N. Impact of Silicone Oil on Free Fatty Acid Particle Formation due to Polysorbate 20 Degradation. Pharm Res 37, 216 (2020). https://doi.org/10.1007/s11095-020-02936-3

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