Cryoprotective agents influence viral dosage and thermal stability of inhalable dry powder vaccines

Manser, Myla; Feng, Xueya; Xing, Zhou; Cranston, Emily D.; Thompson, Michael R.

doi:10.1016/j.ijpharm.2022.121602

Cited by 4 publications

(6 citation statements)

References 39 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A recombinant human serotype 5 adenovirus expressing Green Fluorescent Protein (AdHu5) was prepared in the McMaster University Immunology Research Centre vector facility and was modified to be replication deficient. Upon purification, the adenoviral vector stock was stored in a 5% trehalose storage buffer at -80°C and had a titre of 4.4 × 10 8 pfu∕mL [15].…”

Section: Chemical and Biologicsmentioning

confidence: 99%

“…Details and corresponding notations for all five excipient formulations are summarized in Table 1. A volume of 60 µL AdHu5 stock vector with 5% trehalose was added per 100 mg of excipient based on our previous work that improved viral activity retention at high loading via appropriate selection of the cryoprotective storage solution [15]. In all physical characterization and flowability tests described in Section "Thermal Aging", a placebo solution of 5% trehalose was added at 60 µL/100 mg of excipient instead of the viral stock, as particle size and morphology of mannitol-dextran spray dried powder is the same with and without AdHu5 present [16].…”

Section: Sample Preparation and Spray Dryingmentioning

confidence: 99%

“…Viral activity testing of each spray dried powder formulation was tested in vitro by following the same protocol described in our previous work [15]. Briefly, α-MEM was used to reconstitute each spray dried sample to a solids concentration of 1% immediately before infecting 40,000 confluent A-549 cells on a 96-well plate and incubating overnight at 37°C.…”

Section: Viral Activity Testing Via Flowcytometrymentioning

confidence: 99%

“…Each spray dried formulation was stored under accelerated aging conditions to determine the impact of excipient formulation on thermal stability of the encapsulated adenovirus. Using a previously described protocol [15], each powder formulation was stored in a water bath at 45°C for 72 h prior to GFP quantification via flowcytometry. Moisture uptake was prevented during the aging process by storing the spray dried powder within microcentrifuge tubes that were sealed with Parafilm® wax, placed within a plastic bag containing desiccant and added to a sealed glass jar with additional desiccant.…”

Section: Thermal Agingmentioning

confidence: 99%

“…A blend of mannitol and dextran has been previously shown to successfully retain viral activity of AdHu5 through spray drying, in addition to providing thermal stabilization [9,14,15]. Of these previous studies, the most successful formulation blended mannitol and dextran in a 3:1 mass ratio using a moderately low molecular weight dextran of 40 kDa [16].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Dextran Mass Ratio Controls Particle Drying Dynamics in a Thermally Stable Dry Powder Vaccine for Pulmonary Delivery

et al. 2022

Self Cite

View full text Add to dashboard Cite

Purpose Thermally stable, spray dried vaccines targeting respiratory diseases are promising candidates for pulmonary delivery, requiring careful excipient formulation to effectively encapsulate and protect labile biologics. This study investigates the impact of dextran mass ratio and molecular weight on activity retention, thermal stability and aerosol behaviour of a labile adenoviral vector (AdHu5) encapsulated within a spray dried mannitol-dextran blend. Methods Comparing formulations using 40 kDa or 500 kDa dextran at mass ratios of 1:3 and 3:1 mannitol to dextran, in vitro quantification of activity losses and powder flowability was used to assess suitability for inhalation. Results Incorporating mannitol in a 1:3 ratio with 500 kDa dextran reduced viral titre processing losses below 0.5 log and displayed strong thermal stability under accelerated aging conditions. Moisture absorption and agglomeration was higher in dextran-rich formulations, but under low humidity the 1:3 ratio with 500 kDa dextran powder had the lowest mass median aerodynamic diameter (4.4 µm) and 84% emitted dose from an intratracheal dosator, indicating strong aerosol performance. Conclusions Overall, dextran-rich formulations increased viscosity during drying which slowed self-diffusion and favorably hindered viral partitioning at the particle surface. Reducing mannitol content also minimized AdHu5 exclusion from crystalline regions that can force the vector to air-solid interfaces where deactivation occurs. Although increased dextran molecular weight improved activity retention at the 1:3 ratio, it was less influential than the ratio parameter. Improving encapsulation ultimately allows inhalable vaccines to be prepared at higher potency, requiring less powder mass per inhaled dose and higher delivery efficiency.

show abstract

Section: Chemical and Biologicsmentioning

confidence: 99%

Section: Sample Preparation and Spray Dryingmentioning

confidence: 99%

Section: Viral Activity Testing Via Flowcytometrymentioning

confidence: 99%

Section: Thermal Agingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Dextran Mass Ratio Controls Particle Drying Dynamics in a Thermally Stable Dry Powder Vaccine for Pulmonary Delivery

et al. 2022

Self Cite

View full text Add to dashboard Cite

show abstract

Design Considerations for Intratracheal Delivery Devices to Achieve Proof-of-Concept Dry Powder Biopharmaceutical Delivery in Mice

Manser

Jeyanathan

et al. 2023

Pharm Res

Self Cite

View full text Add to dashboard Cite

Purpose Intratracheal delivery and consistent dosing of dry powder vaccines is especially challenging in mice. To address this issue, device design of positive pressure dosators and actuation parameters were assessed for their impacts on powder flowability and in vivo dry powder delivery. Methods A chamber-loading dosator assembled with stainless-steel, polypropylene or polytetrafluoroethylene needle-tips was used to determine optimal actuation parameters. Powder loading methods including tamp-loading, chamber-loading and pipette tip-loading were compared to assess performance of the dosator delivery device in mice. Results Available dose was highest (45%) with a stainless-steel tip loaded with an optimal mass and syringe air volume, primarily due to the ability of this configuration to dissipate static charge. However, this tip encouraged more agglomeration along its flow path in the presence of humidity and was too rigid for intubation of mice compared to a more flexible polypropylene tip. Using optimized actuation parameters, the polypropylene pipette tip-loading dosator achieved an acceptable in vivo emitted dose of 50% in mice. After administering two doses of a spray dried adenovirus encapsulated in mannitol-dextran, high bioactivity was observed in excised mouse lung tissue three days post-infection. Conclusions This proof-of-concept study demonstrates for the first time that intratracheal delivery of a thermally stable, viral-vectored dry powder can achieve equivalent bioactivity to the same powder, reconstituted and delivered intratracheally. This work may guide the design and device selection process for murine intratracheal delivery of dry powder vaccines to help progress this promising area of inhalable therapeutics.

show abstract

Screening amino acid additives as aerosolization modifiers for spray dried inhalable viral-vectored vaccines

Singh

Son

Dolovich

et al. 2022

Journal of Drug Delivery Science and Technology

Self Cite

View full text Add to dashboard Cite

Cryoprotective agents influence viral dosage and thermal stability of inhalable dry powder vaccines

Cited by 4 publications

References 39 publications

Dextran Mass Ratio Controls Particle Drying Dynamics in a Thermally Stable Dry Powder Vaccine for Pulmonary Delivery

Dextran Mass Ratio Controls Particle Drying Dynamics in a Thermally Stable Dry Powder Vaccine for Pulmonary Delivery

Design Considerations for Intratracheal Delivery Devices to Achieve Proof-of-Concept Dry Powder Biopharmaceutical Delivery in Mice

Screening amino acid additives as aerosolization modifiers for spray dried inhalable viral-vectored vaccines

Contact Info

Product

Resources

About