Real-world data (RWD) and real-world evidence (RWE) are valuable resources for drug development strategies. Historically, it has been used for safety evaluation during post-marketing surveillance. RWD and RWE have been utilized in the regulatory decision-making process for drug effectiveness, especially for rare diseases and cancers, where conducting randomized controlled trials is challenging. The Food and Drug Administration (FDA) is actively working on providing trustworthy information derived from RWD and RWE to supplement the data from clinical trials. This review discusses the potential use of RWE to make regulatory decisions on drug effectiveness for certain therapeutic areas as well as the challenges in drawing inferences on drug effectiveness from RWE. A review of FDA-approved new drug applications and biologics license applications suggests that several methodological considerations should be deliberated when designing a study using RWE to demonstrate product effectiveness. The acceptance of RWE, while promising, is dependent on the relevance and reliability of the data. The insight and engagement of all stakeholders contribute to the successful use of RWE for clinical evaluations.

Nitrosamines drug substance related impurities (NDSRI) are organic impurities, highly potent mutagenic substances that are classified as human carcinogens. Regulatory guidance's in this area provide procedures for control and risk mitigation of NDSRI's in drug products. While efforts are made to control the NDSRI's, cases where NDSRI's are observed post- pivotal bioequivalence studies may require additional bioequivalence studies, between pre- and post- change formulations. The recent USFDA (United State Food and Drug Administration) guidance on "Control of Nitrosamine Impurities in Human Drugs" provides alternative BE methodologies for biopharmaceutics classification system (BCS) I, II and III drugs based on in vitro testing. For BCS IV molecule containingimmediate release (IR) formulations, physiologically based pharmacokinetic (PBPK) modeling approach is recommended. In this context, this present article discusses use of PBPK modeling approaches as alternative BE methodologies for BCS IV molecule containingIR products. We have summarized use of in-house in silico tool for early risk prediction of NDSRI's solely based on molecule structure. This tool was used to predict the carcinogenic potential of 37 BCS IV molecules and further clinical exposure risk assessment was conducted on identified 5 high risk category molecules namely edoxaban, selumetinib, bosutinib, furosemide, hydrochlorothiazide where transporters are involved in absorption. The PBPK models were used to evaluate the impact of altered permeability and transporter kinetics on exposures, that may happen in presence of antioxidants. Further, the impact of permeability within ± 10-20% was evaluated on clinical exposures to determine permeability safe space (region within which bioequivalence is guaranteed as compared to the target formulation).

The selective delivery of small molecule compounds such as Gemcitabine to tumor cells is a promising methodology for enhancing therapeutic efficacy and attenuating the side effects of anticancer drugs. Aptamers are useful as target-directed ligands for tumor-selective drug delivery due to their ability to bind specific proteins. However, the drug must be released from the aptamer after the conjugate is taken up by the cell to exert its pharmacological effect. In this study, we designed and synthesized a conjugate in which a linker cleaved by glutathione, which is highly expressed in tumor cells, was inserted between the aptamer (AS1411) and Gemcitabine. Almost all Gemcitabine was released from the conjugate after 30 min in the presence of 6 mM glutathione. AS1411 is known to bind to nucleolin, which is highly expressed on tumor cells. The cytotoxicity of the AS1411 and Gemcitabine conjugate with a disulfide bond on A549 cells was higher than that of the conjugate without a disulfide bond. Furthermore, the cytotoxicity of the disulfide-linked conjugate of AS1411 and Gemcitabine was higher in A549 cells than in MCF10A cells, which were used as the model of normal cells. These results indicate that disulfide conjugation enhanced the tumor cell-selective cytotoxicity of Gemcitabine with AS1411.Graphical

Quality by Design (QbD) is a transformative and systematic approach to developing top-tier pharmaceutical products, ushering in a departure from traditional trial-and-error methods toward a more science-based, risk-oriented, and holistic strategy. Central to QbD implementation is the meticulous development of formulations and manufacturing processes, consistently fulfilling predefined quality objectives. The core objective of QbD remains unwavering — to guarantee the steadfast alignment of the final pharmaceutical product with predetermined quality attributes, thereby mitigating batch-to-batch variations and potential recalls. This article succinctly explores the multifaceted application of QbD methodology within the pharmaceutical industry. Emphasizing its pivotal role in research and development, manufacturing, quality control, and quality assurance, the discussion navigates through the strategic deployment of QbD elements and tools. Amidst the evident advantages of QbD, challenges persist in its widespread adoption within the pharmaceutical sector and regulatory frameworks. This article sheds light on the regulatory landscape that currently governs the implementation of QbD in these crucial stages of pharmaceutical processes. For that reason, this review article aims to provide researchers, scientists, and industry professionals with a thorough introduction to QbD so they may adopt this methodical approach to developing and producing high-quality pharmaceutical products, always in compliance with the underlying regulations.Graphical

Administration of human therapeutic proteins such as monoclonal antibodies (mAb) to animals during preclinical drug development often leads to the development of anti-drug antibodies (ADA). ADA may reduce the systemic exposure of the mAb by enhancing its immune-complex mediated clearance. Thus, ADA may hinder the preclinical pharmacokinetic and toxicology assessments of mAbs. To mitigate this effect, we explored the ability of short-term administration of immunosuppressants to induce prolonged immune tolerance towards a human mAb, erenumab, in rats. In two studies, we investigated dosing regimens using the immunosuppressants methotrexate and tacrolimus/sirolimus combination, and compared them to non-immunosuppressed control groups. Each study comprised three phases: induction (weeks 1–4), washout (weeks 5–8), and rechallenge (weeks 9–12). Animals received mAb during the induction and rechallenge phase, while immunosuppression was limited to the induction and washout phase. Blood samples were collected at predefined time-points for erenumab and ADA quantification. The tacrolimus/sirolimus regimen, but not the tested methotrexate regimens, completely prevented ADA formation in all treated animals relative to the control groups. The tacrolimus/sirolimus treated animals not only remained ADA-negative with initial immunosuppression during the induction phase but remained ADA-negative even after erenumab rechallenge suggesting the induction of immune-tolerance beyond the immunosuppressive treatment period. Correspondingly, erenumab systemic exposures were maintained throughout the study period in all animals in the tacrolimus/sirolimus group and were similar to the erenumab exposures in ADA-negative animals of the control group. In contrast, ADA-positive animals in the control group exhibited a 60–80% reduction in erenumab exposures.Graphical

Accurate dose preparation and administration are critical components in clinical studies to ensure patient safety and drug efficacy. Closed System Transfer Devices (CSTD) are widely used for the preparation and administration of hazardous drugs. Recent literature has raised concerns about potential under- or over-dosing when doses are prepared using CSTD due to their high hold-up volume. To ensure dose accuracy, it is essential to follow proper preparation methods when using CSTD. Seven commonly used CSTD were evaluated for product transfer accuracy from vials to IV bags using four methods: no bag spike flushing, flushing bag spikes with the original syringes, flushing bag spikes with new syringes, and circle priming. The data confirmed that under- and over-dosing occurred frequently at transfer volume ≤ 2mL when not flushing bag spikes or flushing bag spikes with the original syringes as suggested by some manufacturers' instructions. The study also demonstrated that flushing bag spikes with the new syringes or using circle priming is effective in enabling accurate volume transfer from vials to IV bags at transfer volumes ≤ 2mL. Both methods are feasible for implementation in pharmacies or hospitals to ensure dose preparation accuracy using CSTD. It is recommended that clinical sites flush bag spikes with the new syringes or use circle priming method for low volume transfer to ensure dose preparation accuracy. CSTD manufacturers should consider including these methods in their Instructions for Use (IFU).

The pharmacokinetics of antibodies with varied binding kinetics were simulated to assess the role of affinity and binding microconstants (kon, koff) on tumor exposure and intra-tumoral distribution. Anti-HER2 constructs (trastuzumab, pertuzumab, VK3VH6, and conjugates with DM1 and gelonin) were produced, purified, and tested for binding and cytotoxicity in vitro, and for intra-tumoral distribution and anti-tumor efficacy in mice. Simulations demonstrated that homogeneity in intra-tumoral distribution increases with increases in koff and with decreases in kon. Interestingly, simulations also predicted that homogeneity in tumor distribution may be improved by decreasing kon and koff in parallel (without changing affinity). Relative to trastuzumab, pertuzumab exhibits similar affinity but a ~ fivefold smaller kon and koff, while VK3VH6 exhibits a similar koff but a ~ 30-fold lower kon and affinity. Conjugate concentrations associated with 50% inhibition of cell proliferation (IC50s) were found to vary with affinity, where IC50 values were similar for pertuzumab and trastuzumab, and higher for VK3VH6. Consistent with model simulations, VK3VH6 and pertuzumab demonstrated more homogeneous tumor distribution than trastuzumab. Although treatment differences were not statistically significant, pertuzumab and VK3VH6 conjugates showed trends for increased survival time relative to mice treated with trastuzumab conjugates. Our simulation and experimental results demonstrate complex relationships between antibody-antigen binding kinetics, intratumoral distribution, and efficacy. The rate constant of association, kon, is an underappreciated determinant of intra-tumoral distribution; among high-affinity antibodies, those with lower values of kon may be expected to exhibit improved intra-tumoral distribution and, potentially, efficacy.Graphical