Skip to main content

Advertisement

SpringerLink
Log in

Quality-by-Design Approach for Biological API Encapsulation into Polymersomes Using “Off-the-Shelf” Materials: a Study on L-Asparaginase

  • Research Article
  • Published:
AAPS PharmSciTech Aims and scope Submit manuscript

Abstract

Polymersomes are versatile nanostructures for protein delivery with hydrophilic core suitable for large biomolecule encapsulation and protective stable corona. Nonetheless, pharmaceutical products based on polymersomes are not available in the market, yet. Here, using commercially available copolymers, we investigated the encapsulation of the active pharmaceutical ingredient (API) L-asparaginase, an enzyme used to treat acute lymphoblastic leukemia, in polymersomes through a quality-by-design (QbD) approach. This allows for streamlining of processes required for improved bioavailability and pharmaceutical activity. Polymersomes were prepared by bottom-up (temperature switch) and top-down (film hydration) methods employing the diblock copolymers poly(ethylene oxide)–poly(lactic acid) (PEG45-PLA69, PEG114-PLA153, and PEG114-PLA180) and the triblock Pluronic® L-121 (poly(ethylene oxide)–poly(propylene oxide)–poly(ethylene oxide), PEG5-PPO68-PEG5). Quality Target Product Profile (QTPP), Critical Quality Attributes (CQAs), Critical Process Parameters (CPPs), and the risk assessment were discussed for the early phase of polymersome development. An Ishikawa diagram was elaborated focusing on analytical methods, raw materials, and processes for polymersome preparation and L-asparaginase encapsulation. PEG-PLA resulted in diluted polymersomes systems. Nonetheless, a much higher yield of Pluronic® L-121 polymersomes of 200 nm were produced by temperature switch, reaching 5% encapsulation efficiency. Based on these results, a risk estimation matrix was created for an initial risk assessment, which can help in the future development of other polymersome systems with biological APIs nanoencapsulated.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Ali U, Naveed M, Ullah A, Ali K, Shah SA, Fahad S, et al. L-Asparaginase as a critical component to combat acute lymphoblastic leukaemia (ALL): a novel approach to target ALL. Eur J Pharmacol. 2016;771:199–210.

    Article  CAS  PubMed  Google Scholar 

  2. Apolinário AC, Almeida Pachioni-Vasconcelos J, Pessoa A, Rangel-Yagui CO. Polymersomes versus liposomes: the “magic bullet” evolution. Quim Nova. 2017;4(7):810–7.

    Google Scholar 

  3. Apolinário AC, Magoń MS, Pessoa A, Rangel-Yagui CO. Challenges for the self-assembly of poly(ethylene glycol)-poly(lactic acid) (PEG-PLA) into polymersomes: beyond the theoretical paradigms. Nanomaterials. 2018;8(6):1–16.

    Article  CAS  Google Scholar 

  4. Bartenstein JE, Robertson J, Battaglia G, Briscoe WH. Stability of polymersomes prepared by size exclusion chromatography and extrusion. Colloids Surfaces A Physicochem Eng Asp. 2016;506:739–46.

    Article  CAS  Google Scholar 

  5. Blackman LD, Varlas S, Arno MC, Houston ZH, Fletcher NL, Thurecht KJ, et al. Confinement of therapeutic enzymes in selectively permeable polymer vesicles by polymerization-induced self-assembly (PISA) reduces antibody binding and proteolytic susceptibility. ACS Cent Sci. 2018;4(6):718–23.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Blanco E, Shen H, Ferrari M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat Biotechnol. 2015;33(9):941–51.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Bleul R, Thiermann R, Maskos M. Techniques to control polymersome size. Macromolecules. 2015;48(20):7396–409.

    Article  CAS  Google Scholar 

  8. Cheng X, Kim JK, Kim Y, Bowie JU, Im W. Molecular dynamics simulation strategies for protein-micelle complexes. Biochim Biophys Acta Biomembr. 2016;1858(7):1566–72.

    Article  CAS  Google Scholar 

  9. Contini C, Pearson R, Wang L, Rizzello L, Ruiz-perez L, Contini C, et al. Bottom-up evolution of vesicles from disks to bottom-up evolution of vesicles from disks to high-genus polymersomes. iScience. 2018;7:132–44.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cruz MEM, Gaspar MM. Liposomal L-asparaginase: in vitro evaluation. Int J Pharm. 1993;96(1–3):67–77.

    Article  CAS  Google Scholar 

  11. Danaei M, Dehghankhold M, Ataei S, Davarani FH, Javanmard R, Dokhani A, et al. Impact of particle size and polydispersity index on the clinical applications of lipidic nanocarrier systems. Pharmaceutics. 2018;10(2):1–17.

    Article  CAS  Google Scholar 

  12. David P, Morbidelli M. Process for protein PEGylation. J Control Release. 2014;180:134–49.

    Article  CAS  Google Scholar 

  13. Dionzou M, Morère A, Roux C, Lonetti B, Marty J-D, Mingotaud C, et al. Comparison of methods for the fabrication and the characterization of polymer self-assemblies: what are the important parameters? Soft Matter. 2016;12(7):2166–76.

    Article  CAS  PubMed  Google Scholar 

  14. Discher DE, Ahmed F. Polymersomes. Annu Rev Biomed Eng. 2006;8:323–41.

    Article  CAS  PubMed  Google Scholar 

  15. Discher BM, Won YY, Ege DS, Lee JC, Bates FS, Discher DE, et al. Polymersomes: tough vesicles made from diblock copolymers. Science. 1999;284(5417):1143–6.

    Article  CAS  PubMed  Google Scholar 

  16. European Medicines Agency (EMA), 2012. ICH guideline Q11 on development and manufacture of drug substances (chemical entities and biotechnological/biological entities) pp. 29. November 2012. https://www.ema.europa.eu/documents/scientific-guideline/ich-guideline-q11-development-manufacture-drug-substances-chemical-entities-biotechnological/biological-entities_en.pdf (Accessed 10 Sept 2018).

  17. European Medicines Agency (EMA), 2017. ICH guideline Q11 on development and manufacture of drug substances (chemical entities and biotechnological/biological entities)—questions and answers, pp. 20. 1 September 2017. https://www.ema.europa.eu/documents/other/ich-guideline-q11-development-manufacture-drug-substances-chemical-entities-biotechnological/biological-entities-questions-answers-adopted_en.pdf (Accessed 10 Sept 2018).

  18. FDA, 2017. FDA guidance for industry (draft): drug products, including biological products, that contain nanomaterials guidance for industry drug products, including biological products, that contain nanomaterials (December 2017). https://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM588857.pdf (Accessed 1 Oct 2018).

  19. Flühmann B, Ntai I, Brorchard G, Simoens S, Mühlebach S. Nanomedicines: the magic bullets reaching their target? Eur J Pharm Sci. 2018; In press article.

  20. Gaumet M, Vargas A, Gurny R, Delie F. Nanoparticles for drug delivery: the need for precision in reporting particle size parameters. Eur J Pharm Biopharm. 2008;69(1):1–9.

    Article  CAS  PubMed  Google Scholar 

  21. Han H. The effect of nanoparticle size on in vivo pharmacokinetics and cellular interaction. Nanomedicine. 2016;11(6):673–92.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. He C, Hu Y, Yin L, Tang C, Yin C. Effects of particle size and surface charge on cellular uptake and biodistribution of polymeric nanoparticles. Biomaterials. 2010;31(13):3657–66.

    Article  CAS  PubMed  Google Scholar 

  23. Hirst, N.C., 2008. Solvency effects on polymer surfactant interactions. Thesis. Cardiff University Wales, UK.

    Google Scholar 

  24. ICH, 2005. International Conference on Harmonisation of technical requirements for registration of pharmaceuticals for human use. ICH harmonised tripartite guideline: quality risk management Q9 https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q9/Step4/Q9_Guideline.pdf (Accessed 15 Oct 2018).

  25. ICH, 2008. International Conference on Harmonisation of technical requirements for registration of pharmaceuticals for human use. ICH harmonised tripartite guideline: pharmaceutical quality systems (Q10) https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q10/Step4/Q10_Guideline.pdf (Accessed 15 Oct 2018).

  26. ICH, 2009. International Conference on Harmonisation of technical requirements for registration of pharmaceuticals for human use. ICH harmonised tripartite guideline: pharmaceutical development Q8(R2) https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q8_R1/Step4/Q8_R2_Guideline.pdf (Accessed 15 Oct 2018).

  27. Joseph A, Contini C, Cecchin D, Nyberg S, Ruiz-perez L, Gaitzsch J, et al. Chemotactic synthetic vesicles: design and applications in blood-brain barrier crossing. Sci Adv. 2017;3(8):1–3.

    Article  CAS  Google Scholar 

  28. Li F, Haan LHJ, Marcelis ATM, Leermakers FAM, Cohen Stuart MA, Sudhölter EJR, et al. Pluronic polymersomes stabilized by core cross-linked polymer micelles. Soft Matter. 2009;5(20):4042–6.

    Article  CAS  Google Scholar 

  29. Maruyama T, Izaki S, Kurinomaru T, Handa K, Kimoto T, Shiraki K. Protein-poly (amino acid) precipitation stabilizes a therapeutic protein L-asparaginase against physicochemical stress. J Biosci Bioeng. 2015;120(6):720–4.

    Article  CAS  PubMed  Google Scholar 

  30. Messager L, Gaitzsch J, Chierico L, Battaglia G. Novel aspects of encapsulation and delivery using polymersomes. Curr Opin Pharmacol. 2014;18:104–11.

    Article  CAS  PubMed  Google Scholar 

  31. Paliwal R, Babu RJ, Palakurthi S. Nanomedicine scale-up technologies: feasibilities and challenges. AAPS PharmSciTech. 2014;15(6):1527–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Pallagi E, Ambrus R, Szabó-révész P, Csóka I. Adaptation of the quality by design concept in early pharmaceutical development of an intranasal nanosized formulation. Int J Pharm. 2015;491(1–2):384–92.

    Article  CAS  PubMed  Google Scholar 

  33. Pallagi E, Ismail R, Paál TL, Csóka I. Initial risk assessment as part of the quality by design in peptide drug containing formulation development. Eur J Pharm Sci. 2018;122:160–9.

    Article  CAS  PubMed  Google Scholar 

  34. Pelaz B, Del Pino P, Maffre P, Hartmann R, Gallego M, Rivera-Fernández S, et al. Surface functionalization of nanoparticles with polyethylene glycol: effects on protein adsorption and cellular uptake. ACS Nano. 2015;9(7):6996–7008.

    Article  CAS  PubMed  Google Scholar 

  35. Place AE, Stevenson KE, Vrooman LM, Harris MH, Hunt SK, Brien JEO, et al. Intravenous pegylated asparaginase versus intramuscular native Escherichia coli L-asparaginase in newly diagnosed childhood acute lymphoblastic leukaemia (DFCI 05-001): a randomised , open-label phase 3 trial. Lancet. 2015;16(16):1677–90.

    Article  CAS  PubMed  Google Scholar 

  36. Qi H, Zhou H, Tang Q, Lee JY, Fan Z, Kim S, et al. Block copolymer crystalsomes with an ultrathin shell to extend blood circulation time. Nat Commun. 2018;9:1–10.

    Article  CAS  Google Scholar 

  37. Rodríguez-García R, Mell M, López-Montero I, Netzel J, Hellweg T, Monroy F. Polymersomes: smart vesicles of tunable rigidity and permeability. Soft Matter. 2011;7(4):1532.

    Article  CAS  Google Scholar 

  38. Shrivastava A, Arif A, Khurshid M, Abul M, Jain SK, Singhal PK. Recent developments in l-asparaginase discovery and its potential as anticancer agent. Crit Rev Oncol/Hematol. 2016;100(1–10):1–10.

    Article  Google Scholar 

  39. Simões A, Veiga F, Figueiras A, Vitorino C. A practical framework for implementing quality by design to the development of topical drug products: nanosystem-based dosage forms. Int J Pharm. 2018;548(1):385–99.

    Article  PubMed  CAS  Google Scholar 

  40. Sueyoshi D, Anraku Y, Komatsu T, Urano Y, Kataoka K. Enzyme-loaded polyion complex vesicles as in vivo nanoreactors working sustainably under the blood circulation: characterization and functional evaluation. Biomacromolecules. 2017;18(4):1189–96.

    Article  CAS  PubMed  Google Scholar 

  41. Tantra R, Schulze P, Quincey P. Effect of nanoparticle concentration on zeta-potential measurement results and reproducibility. Particuology. 2010;8(3):279–85.

    Article  CAS  Google Scholar 

  42. Wang L, Chierico L, Little D, Patikarnmonthon N, Yang Z, Azzouz M, et al. Encapsulation of biomacromolecules within polymersomes by electroporation. Angew Chemie Int Ed. 2012;51(44):11122–5.

    Article  CAS  Google Scholar 

  43. Wolf M, Wirth M, Pittner F, Gabor F. Stabilisation and determination of the biological activity of l-asparaginase in poly(d,l-lactide-co-glycolide) nanospheres. Int J Pharm. 2003;256(1–2):141–52.

    Article  CAS  PubMed  Google Scholar 

  44. Wong CK, Laos AJ, Soeriyadi AH, Wiedenmann J, Curmi PMG, Gooding JJ, et al. Polymersomes prepared from thermoresponsive fluorescent protein-polymer bioconjugates: capture of and report on drug and protein payloads. Angew Chemie Int Ed. 2015;54(18):5317–22.

    Article  CAS  Google Scholar 

  45. Xu X, Costa AP, Khan MA, Burgess DJ. Application of quality by design to formulation and processing of protein liposomes. Int J Pharm. 2012;434(1–2):349–59.

    Article  CAS  PubMed  Google Scholar 

  46. Yu LX, Amidon G, Khan MA, Hoag SW, Polli J, Raju GK, et al. Understanding pharmaceutical quality by design. AAPS J. 2014;16(4):771–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Zelikin AN, Ehrhardt C, Healy AM. Materials and methods for delivery of biological drugs. Nat Chem. 2016;8(11):997–1007.

    Article  CAS  PubMed  Google Scholar 

  48. Robertson JD, Yealland G, Avila-Olias M, Chierico L, Bandmann O, Renshaw S A, et al. pH-sensitive tubular polymersomes: formation and applications in cellular delivery. ACS Nano. 2014;8:4650–4661

  49. Meglič HS, Kotnik T. Electroporation-Based Applications in Biotechnology. In: Miklavčič D. (eds) Handbook of Electroporation. Springer: Cham; 2017. p. 2153–2169

Download references

Acknowledgments

We acknowledge support from the State of São Paulo Research Foundation (FAPESP-Brazil) projects 2013/08617-7 (Thematic project), 2014/10456-4 and 2017/03811-0 (Apolinário, A.C. PhD fellowships), and 2016/03887-4 (Oliveira, C.A. Post-Doctoral Fellowship), Coordination for the Improvement of Higher Education Personnel (CAPES, Project 001), and the National Council for Scientific and Technological Development (CNPq-Brazil, project 303334/2014-2). We are in debt with Dr. Monika S. Magón for the enlightening discussions and text reading. Additionally, we thank the BASF Brazil for Pluronic® L-121 donations.

Author information

Authors and Affiliations

  1. Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 580-Bl.16, São Paulo, CEP 05508-000, Brazil

    Alexsandra Conceição Apolinário, Rafael Bertelli Ferraro, Camila Areias de Oliveira, Adalberto Pessoa Jr & Carlota de Oliveira Rangel-Yagui

Authors
  1. Alexsandra Conceição Apolinário
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rafael Bertelli Ferraro
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Camila Areias de Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Adalberto Pessoa Jr
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carlota de Oliveira Rangel-Yagui
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carlota de Oliveira Rangel-Yagui.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(DOCX 290 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Apolinário, A.C., Ferraro, R.B., de Oliveira, C.A. et al. Quality-by-Design Approach for Biological API Encapsulation into Polymersomes Using “Off-the-Shelf” Materials: a Study on L-Asparaginase. AAPS PharmSciTech 20, 251 (2019). https://doi.org/10.1208/s12249-019-1465-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1208/s12249-019-1465-1

KEY WORDS

Search

Navigation