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Identification of Factors of Importance for Spray Drying of Small Interfering RNA-Loaded Lipidoid-Polymer Hybrid Nanoparticles for Inhalation

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Abstract

Background

With the recent approval of the first small interfering RNA (siRNA) therapeutic formulated as nanoparticles, there is increased incentive for establishing the factors of importance for the design of stable solid dosage forms of such complex nanomedicines.

Methods

The aims of this study were: (i) to identify factors of importance for the design of spray-dried siRNA-loaded lipidoid-poly(DL-lactic-co-glycolic acid) hybrid nanoparticles (LPNs), and (ii) to evaluate their influence on the resulting powders by using a quality-by-design approach. Critical formulation and process parameters were linked to critical quality attributes (CQAs) using design of experiments, and an optimal operating space (OOS) was identified.

Results

A series of CQAs were identified based on the quality target product profile. The loading (ratio of LPNs to the total solid content) and the feedstock concentration were determined as critical parameters, which were optimized systematically. Mannitol was chosen as stabilizing excipient due to the low water content of the resulting powders. The loading negatively affected the colloidal stability of the LPNs, whereas feedstock concentration correlated positively with the powder particle size. The optimal mannitol-based solid formulation, defined from the OOS, displayed a loading of 5% (w/w), mass median aerodynamic diameter of 3.3 ± 0.2 μm, yield of 60.6 ± 6.6%, and a size ratio of 1.15 ± 0.03. Dispersed micro-embedded LPNs had preserved physicochemical characteristics as well as in vitro siRNA release profile and gene silencing, as compared to non-spray-dried LPNs.

Conclusion

The optimal solid dosage forms represent robust formulations suitable for higher scale-up manufacturing.

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Abbreviations

ACT:

β-actin

CCF:

Central composite face

CFP:

Critical formulation parameter

COPD:

Chronic obstructive pulmonary disease

CP:

Crossing point

CPP:

Critical processing parameter

CQAs:

Critical quality attributes

DEPC:

Diethyl pyrocarbonate

DoE:

Design of experiments

FBS:

Fetal bovine serum

GUS:

β-glucuronidase

hATTR:

Hereditary transthyretin-mediated amyloidosis

HD:

Heparin and detergent

LPN:

Lipidoid-polymer hybrid nanoparticle

LPS:

Lipopolysaccharide

MLR:

Multiple linear regression

MMAD:

Mass median aerodynamic diameter

OFAT:

One-factor-at-a-time

OG:

Octyl β-D-glucopyranoside

OOS:

Optimal operating space

PDI:

Polydispersity index

PLGA:

Poly(DL-lactic-co-glycolic acid)

PVA:

Polyvinylalcohol

QbD:

Quality-by-design approach

QTPP:

Quality target product profile

SEM:

Scanning electron microscopy

siRNA:

Small interfering RNA

Tg :

Glass transition temperature

TNF:

Tumor necrosis factor

TTR:

Transthyretin

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Acknowledgments and Disclosures

We gratefully acknowledge financial support from the Novo Nordisk Foundation - Denmark (Grant No. NNF17OC0026526), Hørslev-Fonden – Denmark, the Lundbeck Foundation – Denmark (Grant No. R219–2016-908) and Independent Research Fund Denmark (Grant No. DFF-4184-00422). We are also grateful to the Innovative Medicines Initiative Joint Undertaking under grant agreement No.115363 resources which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007–2013) and EFPIA companies’ in kind contribution. This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement No. 600207. We acknowledge the Danish Agency for Science, Technology and Innovation for funding the Zetasizer Nano ZS. We also acknowledge the Alfred Benzon Foundation and Drug Research Academy for co-funding the FLUOstar OPTIMA plate reader and Drug Research Academy for funding the Small-Scale Powder Disperser. Novo Nordisk has kindly supplied the Aerodynamic Particle Sizer Spectrometer 3321. The funding sources had no involvement in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; nor in the decision to submit the paper for publication. We are grateful to Emily Falkenberg for synthesizing and purifying L5 and to Xianghui Zeng for technical assistance. The authors report no potential conflicts.

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Author notes

  1. Cypriane Dormenval and Abhijeet Lokras contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100, Copenhagen Ø, Denmark

    Cypriane Dormenval, Abhijeet Lokras, Guillermo Cano-Garcia, Abishek Wadhwa, Kaushik Thanki, Fabrice Rose, Aneesh Thakur & Camilla Foged

  2. Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, CH-1015, Lausanne, Switzerland

    Cypriane Dormenval

  3. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Jagtvej 162, DK-2100, Copenhagen Ø, Denmark

    Henrik Franzyk

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Correspondence to Camilla Foged.

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Dormenval, C., Lokras, A., Cano-Garcia, G. et al. Identification of Factors of Importance for Spray Drying of Small Interfering RNA-Loaded Lipidoid-Polymer Hybrid Nanoparticles for Inhalation. Pharm Res 36, 142 (2019). https://doi.org/10.1007/s11095-019-2663-y

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