Pharmaceutical Product Development (United States)

Developing a stabilized spray-dried formulation for proteins is one key approach to extending the stability of a dehydrated drug product and providing a broad range of different drug delivery applications. Trehalose has been extensively studied as an excipient to preserve the protein stability in spray-dried formulations. However, the hygroscopic nature of amorphous trehalose makes it prone to recrystallization upon exposure to high temperature and humidity conditions, which can be detrimental to protein stability in the solid state. Herein, we report a formulation approach with the use of 2-hydroxypropyl-β-cyclodextrin (HPβCD) in combination with trehalose to enhance the physical stability of spray-dried solids by inhibiting or delaying the recrystallization of trehalose. Specifically, the effect of HPβCD and trehalose as an excipient alone or the combination of both excipients in stabilizing a model therapeutic monoclonal antibody (mAb1) in a spray-dried formulation has been evaluated at 25 °C/60% relative humidity (RH) and 40 °C/75% RH over 4 weeks. In the solid state, HPβCD can inhibit trehalose recrystallization of spray-dried solids exposed to stress conditions of higher temperatures and humidity. The recrystallization tendency of trehalose was found to be dependent on the protein-to-excipient mass ratios. At a 1:1 protein-to-excipient mass ratio, trehalose is insufficient to ensure adequate protein stability after stress under high humidity, which leads to a significant change in conformational stability and the highest degree of subvisible particle formation for mAb1, primarily due to trehalose recrystallization and high-temperature stresses, while the combination of HPβCD and trehalose has resulted in improved protein stability with a reduction in aggregation propensity. These results show that a combination of HPβCD and trehalose is a viable approach in mitigating trehalose recrystallization and maintaining protein stability of mAbs in spray-dried formulations.

BackgroundRespiratory syncytial virus (RSV) is a leading cause of lower respiratory tract disease among infants and young children worldwide, especially in low- and middle-income countries (LMICs). RSM01 is a novel, highly potent, half-life-extended anti-RSV monoclonal antibody (mAb) candidate primarily being developed for LMICs. Here we present the preclinical characterisation and results of a phase 1 trial of RSM01.MethodsPreclinical characterisation of RSM01 was conducted using in-vitro neutralization assays and cotton rat models.In the first-in-human, double-blind, phase 1 trial, 56 healthy adults were randomised 6:1 within dose cohorts to receive a single dose of RSM01 (n = 48) or placebo (n = 8): 300 mg intravenously (IV), 300 mg intramuscularly (IM) or 1000 mg IV (parallel cohorts), 3000 mg IV, and an expansion cohort of 600 mg IM. Systemic solicited adverse events (AEs) were assessed through day 7; unsolicited AEs were collected through day 151. Pharmacokinetics and anti-drug antibodies (ADA) to RSM01 were assessed using immunoassays. A population pharmacokinetics model predicted paediatric pharmacokinetics parameters using allometric scaling and age-specific population weight statistics of North American and African infants.ResultsRSM01 exhibited highly potent neutralizing activity in the single ng/mL range (0.7–6.4) against diverse RSV-A and RSV-B isolates in vitro. RSM01 also demonstrated prophylactic efficacy in cotton rat models with both RSV subtypes. In the phase 1 clinical trial, the most common unsolicited AEs were COVID-19 (2/48), headache (2/48), and nausea (2/48), all in RSM01-treated participants. The only systemic solicited AEs reported were headache (5/48) and tiredness (2/48) in participants receiving RSM01. No serious AEs or deaths were reported. The half-life of RSM01 was 78 days with dose-proportional increases in Tmax and AUClast after IV administration. Among RSM01-treated participants, 2/48 were ADA positive at baseline, and 1/48 seroconverted to ADA-positive post-baseline.ConclusionsRSM01 is a highly potent, half-life-extended, RSV-neutralising mAb candidate that was shown to be well tolerated in healthy adults. The rate of ADA to RSM01 was low. The long half-life of RSM01 and pharmacokinetics profile support further development of RSM01 as a potential single dose per season prophylaxis to prevent RSV disease in infants.Trial registrationClinicaltrials.gov NCT05118386, Nov 12, 2021.

The urgency and importance of organizing a global effort to harmonize clinical assay validation specific to the vaccine industry was identified during the drafting of the 2020 White Paper in Bioanalysis due to the lack of clarity and regulatory guidance/guidelines in vaccine immunoassay validation. Indeed, the Workshop on Recent Issues in Bioanalysis (WRIB) issues the White Paper in Bioanalysis yearly, which is one of the high-profile articles of the Bioanalysis Journal focused on detailed discussions and recommendations on vaccine assay validation. Since 2017, participation in the WRIB working groups by vaccine assay validation experts and regulators has rapidly increased due to its unique format where industry leaders and regulators can meet and exchange ideas on topics of interest to both groups. In early 2021, Vaccine manufacturers approached WRIB for sponsoring/supporting the authorship and publication of an overarching vaccine assay validation document based on the 2017-2020 discussions and consensus starting from immunogenicity assays first and followed by future papers on molecular and cell-based assay validation. Using industry and WRIB vaccine network, a vaccine immunogenicity assay validation working group was assembled consisting of 16 companies. The work on the first white paper started officially in April 2021 focusing on Vaccine LBA Validation (Part 1), and the drafting of Vaccine LBA Development (Part 2) and Vaccine LBA Monitoring & Transfer (Part 3) are presently ongoing and expected to be published shortly after this paper. Moreover, recommendations on Vaccine Cell-Based Assays Validation (ELISpot and Flow cytometry) and Vaccine Molecular Assays Validation (PCR, NGS, NanoString) are also on the WRIB publications agenda and the drafting is planned to start in mid-2024. For too long, vaccine scientists have not had a clear validation guidance for clinical vaccine immunogenicity assays. We hope that this common effort will help close this regulatory gap.

To identify vaginal morphology and position factors associated with prolapse recurrence following vaginal surgery. Secondary analysis of magnetic resonance images (MRI) of the Defining Mechanisms of Anterior Vaginal Wall Descent cross-sectional study. Eight clinical sites in the US Pelvic Floor Disorders Network. Women who underwent vaginal mesh hysteropexy (hysteropexy) with sacrospinous fixation or vaginal hysterectomy with uterosacral ligament suspension (hysterectomy) for uterovaginal prolapse between April 2013 and February 2015. The MRI (rest, strain) obtained 30-42 months after surgery, or earlier for participants with recurrence who desired reoperation before 30 months, were analysed. MRI-based prolapse recurrence was defined as prolapse beyond the hymen at strain on MRI. Vaginal segmentations (at rest) were used to create three-dimensional models placed in a morphometry algorithm to quantify and compare vaginal morphology (angulation, dimensions) and position. Vaginal angulation (upper, lower and upper-lower vaginal angles in the sagittal and coronal plane), dimensions (length, maximum transverse width, surface area, volume) and position (apex, mid-vagina) at rest. Of the 82 women analysed, 12/41 (29%) in the hysteropexy group and 22/41 (54%) in the hysterectomy group had prolapse recurrence. After hysteropexy, women with recurrence had a more laterally deviated upper vagina (p = 0.02) at rest than women with successful surgery. After hysterectomy, women with recurrence had a more inferiorly (lower) positioned vaginal apex (p = 0.01) and mid-vagina (p = 0.01) at rest than women with successful surgery. Vaginal angulation and position were associated with prolapse recurrence and suggestive of vaginal support mechanisms related to surgical technique and potential unaddressed anatomical defects. Future prospective studies in women before and after prolapse surgery may distinguish these two factors.

Identifying a subgroup of patients who may have an enhanced treatment effect in a randomized clinical trial has received increasing attention recently. For time-to-event outcomes, it is a challenge to define the effectiveness of a treatment and to choose a cutoff time point for identifying subgroup membership, especially in trials in which the two treatment arms do not differ in overall survival. In this paper, we propose a mixture cure model to identify a subgroup for a new treatment that was compared to a classical treatment (or placebo) in a randomized clinical trial with respect to survival time. Using the concordance probability measurement (K-index), we propose a statistic to test the existence of subgroups with effective treatments in the treatment arm. Subsequently, the subgroup is defined by a limited number of covariates based on the estimated area under the curve (AUC). The performance of this method in different scenarios is assessed through simulation studies. A real data example is also provided for illustration.

<strong>ABSTRACT</strong> <strong>Background</strong>: The Basic Health Care Provision Fund (BHCPF) is expected to fund the delivery of a minimum package of healthcare services to Nigerians especially the poor and vulnerable. <strong>Objectives</strong>: This study was designed to commence the assessment of the progress of the processes of the implementation of the fund, and the challenges encountered by operators of the Primary Health Care (PHC) system in Southeast. <strong>Materials and Methods</strong>: This survey conducted between February-August, 2020 entailed recruitment, training of independent investigators, and deployment of tracking survey tools to independent monitors and managers of the BHCPF of the States including the PHC Agencies and Facility Managers. Results were collated and validated in a virtual meeting of public and independent monitors. The study focused on the PHC Facilities in the Southeast States of Nigeria. <strong>Results</strong>: The findings indicate a shaky and delayed start of implementation of the BHCPF in five surveyed States, mainly due to a collection of reasons including non-compliance to processes of the operational guideline; inadequate number, and mix crop of Human Resources for Health (HRH); Inadequate management capacity of supervising agencies to ensure adherence to the guidelines; poor management capacities of Officers in Charge (OICs); conflicts between Ward Development Committees (WDCs) and OICs; extremely low-level awareness of the availability of BHCPF in the communities of the States surveyed <strong>Conclusion</strong>: These issues arising from the take-off and implementation of BHCPF warrant continuing capacity building and supervision of the programme. Certain States are at lower readiness to access the fund due to the alluded complicated processes.