PROJECT SUMMARY We have developed a continuous processing system for nanoparticles with a primary emphasis on liposomes (FDA Grant Award 1U01FD005773), that is designed to allow rapid and high-throughput manufacturing of these critical drug products on-demand in the US. Such a system is particularly crucial given the real and anticipated shortages of drug products with the current pandemic. Our system is currently being fabricated as a cGMP-ready manufacturing skid and qualification with doxorubicin-loaded liposomes will be performed summer 2020 in a cGMP facility. We are seeking to further enhance our cGMP-ready system in two critical areas: (1) develop real- time-release testing methods to incorporate as new process analytical technology (PAT) tools; and (2) evaluate morphological changes of drug-loaded liposomes (e.g. doxorubicin, daunorubicin and vincristine). With respect to the first critical area, we have already identified three PAT tools and plan to further evaluate these to allow incorporation of models with very low error that will be suitable for real-time-release testing. In addition, we propose to evaluate additional PAT at critical areas (e.g. inline Raman or NIR to assess internal structure of drug-loaded liposomes in real-time, as well as spatially-resolved dynamic light scattering for high resolution particle size and size distribution measurement). With respect to the second critical area, morphological differences in liposomal structure that can occur after drug loading may increase the likelihood of adverse clinical events. E.g. doxorubicin loaded liposomes can form elongated nanorods, causing shape change from spheres to ellipsoids – potentially resulting in accumulation away from the tumor site, (i.e. at body extremities, which can result in hand-foot-syndrome). An in-depth understanding of processing conditions to control morphology will be of great benefit to the agency. Accordingly, we will evaluate material attributes and processing parameters for antineoplastic drugs with physicochemical characterization and drug release testing. The proposed project is aligned with the FDA Advancing Regulatory Science Plan within the scope of supporting new approaches to improve product manufacturing and quality. The proposed research advances the development of a continuous manufacturing approach that will improve the manufacturing and quality of complex drug products. More specifically, continuous manufacturing of liposomes will be advanced by establishing real- time-release testing methods and thus reducing production time. Regulatory science will be advanced as detailed analysis will be provided on: incorporation of single and multiple antineoplastic drugs into liposomal formulations; and the relationship between processing parameters and liposomal morphology (e.g. drug crystal shape and size). Such information will be critical for new and generic drug products. Note that previously approved generic liposomal doxorubicin products ...