(T1030-03-18) Optimization of Antibody Based Products Stabilized within a Film Matrix for Nasal Administration

Purpose: Recent studies have shown that many patients prescribed an antibody-based product for long-term therapy discontinue treatment within one year [1, 2]. One of the primary reasons for this is the development of immune reactions, which can be caused by product degradation due to poor handling, and storage of these medications. In one study, only 6.7% of patients prescribed an antibody based therapeutic stored the medication within the temperature range recommended by the manufacturer.  In 25% of these cases, products were stored above 25oC or below 0oC for extended periods of time [3, 4]. To address this issue, formulations that support storage of antibody-based products at 25 oC were identified to alleviate patient mishandling & improve compliance. While many therapeutic antibodies are administered by injection, inhalation offers a non-invasive method for delivery of high concentrations of drug to the respiratory tract.  Film-based formulations that support stability under ambient conditions were then characterized and evaluated for their suitability for intranasal delivery.

Methods: Antibodies and enzymes were mixed with excipients (Table 1), dispensed in 1 ml molds and dried under aseptic conditions.  Stability was determined by use of solutions made from rehydrated films in Western blots, ELISA assays or colorimetric enzyme activity kits (Invitrogen, Sigma Aldrich).  Residual moisture content (RMC) of films was determined by Karl Fisher Titration (V10S Volumetric Titrator, Mettler Toledo).  Dissolution studies were conducted in simulated nasal fluid [5] at 37oC. Viscosity at 25oC was assessed using a RV Brookfield Viscometer and SC4-21 spindle. The VP7 Spray Pump obtained from Aptar Pharmaceuticals was used to visually determine each formulation’s ability to be generated as an aerosol or jet.

Results: A formulation (F8) that retained binding affinity of a monoclonal mouse anti-human alpha-1 antitrypsin antibody during the film forming process dissolved at a rate of 5.1 ± 0.9 mg/min and had a RMC of 8.4 ± 1.1%.  Solutions prepared from rehydrated films made with this formulation were viscous (142 ± 1.4 cp) and produced a jet plume during aerosolization.  Formulation F1, capable of stabilizing a antibody and enzymatic activity of horseradish peroxidase conjugated goat anti-mouse IgG secondary antibody at 25oC for 48 hours, had a similar dissolution rate (6.9 ± 0.9 mg/min) and RMC (11.8 ± 2.4%) with a notably lower viscosity (8.5 ± cp).  This preparation formed an aerosol when dispensed from VP7.  F12 stabilized recombinant beta-galactosidase 10 days at 25oC and had a slower dissolution rate (1.3 ± 0.03 mg/min) and higher RMC and viscosities (12.8 ± 2.5% and 171.6 ± 6.5 cp) than other formulations resulting in a jet plume during aerosolization. F7, which maintained 100% binding affinity of a humanized anti-pertussis toxin antibody (hu11E6) and an antibody-enzyme conjugate at 20 oC  for 1 and 3 months respectively had a similar dissolution rate (1.3 ± 0.1 mg/min) and higher RMC (11.8 ± 0.7%) than F12.  Viscosity of this formulation was 121.7 ± 3.6 cp. Other formulations tested had characteristics that fell within the ranges summarized here.

Conclusion: Formulations containing 1.5-3% HPMC were most effective at maintaining stability of antibodies, enzymes, and antibody-enzyme conjugates at 20oC. Studies have demonstrated that there is a balance between the viscosity of the formulation and the stability of the biologics in solid dosage forms, however, the viscosity after reconstitution is an important consideration for nasal and pulmonary administration.  High viscosity formulations that produce jet plumes can strategically target nasal turbinates for intranasal administration of therapeutic agents to protect against infectious disease. Methods to reduce viscosity during rehydration should also be considered for aerosol delivery to lower portions of the respiratory tract.

References: 1. Krämer, J., & Wiendl, H. (2022). What Have Failed, Interrupted, and Withdrawn Antibody Therapies in Multiple Sclerosis Taught Us?. Neurotherapeutics: the journal of the American Society for Experimental NeuroTherapeutics, 19(3), 785–807.
2. Lundahl, M. L. E., Fogli, S., Colavita, P. E., & Scanlan, E. M. (2021). Aggregation of protein therapeutics enhances their immunogenicity: causes and mitigation strategies. RSC chemical biology, 2(4), 1004–1020.
3. Vlieland, N. D., Gardarsdottir, H., Bouvy, M. L., Egberts, T. C., & van den Bemt, B. J. (2016). The majority of patients do not store their biologic disease-modifying antirheumatic drugs within the recommended temperature range. Rheumatology, 55(4), 704-709.
4. Vlieland, N.D., Nejadnik, M.R., Gardarsdottir, H. et al. The Impact of Inadequate Temperature Storage Conditions on Aggregate and Particle Formation in Drugs Containing Tumor Necrosis Factor-Alpha Inhibitors. Pharm Res 35, 42 (2018).
5. Galgatte, U. C., Kumbhar, A. B., & Chaudhari, P. D. (2014). Development of in situ gel for nasal delivery: design, optimization, in vitro and in vivo evaluation. Drug Delivery, 21(1), 62–73.

Acknowledgements: This work was supported by the National Institute of Allergy and Infectious Disease (U01 AI 078045/MAC), a Texas Health Catalyst award from the University of Texas at Austin Dell Medical School (MAC), a Johnson and Johnson Quickfire Challenge Award (MAC) and a Building Our Own Talent (BOOT) Graduate Fellowship from the University of Texas at Austin College of Pharmacy (J.D.).


Table 1. Composition of Formulations


Table 2. Properties of Formulations That Stabilize Antibody Based Products at Ambient Temperature