(M1430-05-27) Self-Emulsifying NanoPODS (Nano-PND Oral Delivery Systems) for Oral Delivery of PND1186: In Silico Modeling for Bioavailability Estimation

Purpose: The treatment regimens for pancreatic ductal adenocarcinoma (PDAC) have witnessed notable advancements, yet the overall efficacy of the treatments remains low, with significant side effects. Focal adhesion kinase (FAK) inhibitors have proven to aid the therapeutic potential of anti-cancer agents. PND1186 (PND) is a FAK inhibitor that disrupts cancer processes such as cell survival and proliferation, cell adhesion and migration, angiogenesis, chemoresistance and tumor microenvironment that reduces metastasis. However, the pharmacological potential of PND is limited by its poor solubility and bioavailability. Poor solubility and rapid precipitation of weakly basic drug in intestine result in poor oral absorption. As a solution, we have developed self-emulsifying Nano-PND oral delivery systems with Soluplus (NanoPODS-S) and without Soluplus (NanoPODS) to deliver PND preconcentrate that emulsifies in an aqueous buffer for enhanced absorption in the intestine. This study evaluates the efficacy of NanoPODS and NanoPODS-S for improved oral bioavailability with better cytotoxicity efficacy on PDAC cell lines. NanoPODS-S is the first of its kind, self-nanoemulsifying system containing a polymeric precipitation inhibitor. It will be a novel platform technology for rapid and enhanced dissolution of weakly basic anticancer molecules from intestine

Methods: The saturation solubility of PND was determined in aqueous buffers at different pH values (1.2,2,3.5,4.5,6.8,7.4) and water, oils, co-solvents, and surfactants. To formulate NanoPODS, a cocktail of oils was prepared by mixing melted Capmul MCM and Captex MCT, with Tween 80 in a ratio of (1:2:2) v/v at 37°C using a vortex mixer. For physiochemical characterization, the particle size and polydispersity index (PDI) were determined using a dynamic light scattering (DLS) particle size analyzer, and the zeta potential was analyzed using laser doppler Microelectrophoresis (NanoZS^®, Malvern Instruments, UK). Several polymers such as HPMC, Soluplus, Parteck MXP, Kollidon VA64, PEG 4000, and Cavasol were screened as a precipitation inhibitor (PPI) by evaluating saturation solubility of NanoPODS preconcentrate in 0.1% w/v polymeric solutions at pH 4.5 and 6.8. Silica-based Co-Processed Excipients (CPEs) were prepared by co-processing F100 and Soluplus or Parteck MXP in a 1:1 weight ratio to optimize a solid carrier for NanoPODS. The in vitro dissolution analysis was conducted using USP (US Pharmacopeia) apparatus II (Symphony 7100 Bathless Dissolution Apparatus, Distek, North Brunswick, NJ) at 50 rpm at 37±0.5°C. The changes in the solid state of PND in the formulation were evaluated by Differential Scanning Calorimetry (DSC) using a Q200 modulated DSC equipment (TA Instruments, New Castle, DE, USA). The optimized formulations were evaluated in vitro for cytotoxicity, cell invasion, migration, and permeability assay. An in-silico simulation was performed using GastroPlus™ simulation software to predict the Physiologically based Pharmacokinetics (PBPK) of free PND and NanoPODS-S.

Results: Optimized NanoPODS depicted a particle size of 133.6±1, PDI of 0.122±0.114, and a surface potential of -1.3±0.700, and >70% PND released at pH 6.8 from NanoPODS-S. Figure1(a) illustrates the steps involved in formulation development of NanoPODS and NanoPODS-S. Figure1(b-e) shows the saturation solubility results for PND in different buffers, surfactants, oils, and co-solvents. As shown in Figure2(a-d), the highest saturation solubility of PND was observed in 0.1% w/v solution of Soluplus, followed by Parteck MXP. Figure2(e)-(f) shows that NanoPODS resuspended in 0.1% w/v polymer solutions demonstrated a droplet size of < 180±3.1 nm and PDI < 0.3±0.04. Figure2(g) shows clear microscopic images of NanoPODS suspended in 0.1% w/v Soluplus solutions rendered free of precipitation. Figure2(h)-(i) summarizes the %PND release evaluated by dissolution studies for different formulation samples. Figure3(a)-(b) shows the cell invasion assay results on MIA PaCa-2 cells and BxPC-3 cells respectively. PND concentrations of 10 µM and 5 µM reduced cell invasion potential by ~13.5 and ~7-fold respectively, as depicted in Figure3(c)-(d). The migration assay results are shown in Figure3(e)-(f). Figure3(g) shows the cellular uptake of NanoPODS and Figure3(h) shows the permeability of NanoPODS-S. Figure3(i) shows that the total fraction of PND absorbed (% Fa) translated as the oral bioavailability of free PND is only 20.18%, while that from NanoPODS-S is 99.99%. Figure 3(j) shows the regional absorption of PND from NanoPODS in different compartments of the small intestine as obtained by in-silico pharmacokinetic modeling.

Conclusion: This is the first study to report enhanced oral solubility of PND with supporting evidence from solubility studies, including dissolution assessment and in-silico pharmacokinetics. The addition of Soluplus in DMA to formulate a self-emulsifying NanoPODS preconcentrate has been studied and reported for the first time as a novel strategy to formulate drug-loaded nanoemulsions. The self-emulsifying NanoPODS-S showed significant enhancement in aqueous solubility without any visible drug precipitation. In-vitro evaluations included the assessment of minimum inhibitory concentration on pancreatic cancer cell lines and permeability assays demonstrating lower IC50 values for NanoPODS-S that confirm increased PND potency in NanoPODS-S. Overall, this report gathers relevant evidence to justify the role of a self-nanoemulsifying strategy for enhanced solubility of poorly soluble PND for maximal therapeutic action.

References: Adamska, A., Domenichini, A., & Falasca, M. (2017). Pancreatic ductal adenocarcinoma: current and evolving therapies. International journal of molecular sciences, 18(7), 1338.
Asong, G. M., Voshavar, C., Amissah, F., Bricker, B., Lamango, N. S., & Ablordeppey, S. Y. (2022). An Evaluation of the Anticancer Properties of SYA014, a Homopiperazine-Oxime Analog of Haloperidol in Triple Negative Breast Cancer Cells. Cancers, 14(24), 6047.
Baloch, J., Sohail, M. F., Sarwar, H. S., Kiani, M. H., Khan, G. M., Jahan, S., Rafay, M., Chaudhry, M. T., Yasinzai, M., & Shahnaz, G. (2019). Self-nanoemulsifying drug delivery system (SNEDDS) for improved oral bioavailability of chlorpromazine: in vitro and in vivo evaluation. Medicina, 55(5), 210.
Bekkali, N. L., & Oppong, K. W. (2017). Pancreatic ductal adenocarcinoma epidemiology and risk assessment: Could we prevent? Possibility for an early diagnosis. Endoscopic ultrasound, 6(Suppl 3), S58.
Berthelsen, R., Sjögren, E., Jacobsen, J., Kristensen, J., Holm, R., Abrahamsson, B., & Müllertz, A. (2014). Combining in vitro and in silico methods for better prediction of surfactant effects on the absorption of poorly water soluble drugs—A fenofibrate case example. International journal of pharmaceutics, 473(1-2), 356-365.
Daware, S., Patki, M., Saraswat, A., Palekar, S., & Patel, K. (2022). Development of a safe pediatric liquisolid self-nanoemulsifying system of triclabendazole for the treatment of fascioliasis. International journal of pharmaceutics, 626, 122163.
Fu, Y., Rathod, D., & Patel, K. (2020). Protein kinase C inhibitor anchored BRD4 PROTAC PEGylated nanoliposomes for the treatment of vemurafenib-resistant melanoma. Experimental Cell Research, 396(1), 112275.
Gupta, A., Eral, H. B., Hatton, T. A., & Doyle, P. S. (2016). Nanoemulsions: formation, properties and applications. Soft matter, 12(11), 2826-2841.
Han, H., Li, Y., Peng, Z., Long, K., Zheng, C., Wang, W., Webster, T. J., & Ge, L. (2020). A Soluplus/Poloxamer 407-based self-nanoemulsifying drug delivery system for the weakly basic drug carvedilol to improve its bioavailability. Nanomedicine: Nanotechnology, Biology and Medicine, 27, 102199.
Harwood, M. D., Zettl, K., Weinheimer, M., Pilla-Reddy, V., Shen, H., Jacobs, F., Chu, X., Huth, F., Nakakariya, M., & Chothe, P. P. (2023). Interlaboratory Variability in the Madin–Darby Canine Kidney Cell Proteome. Molecular Pharmaceutics.

Acknowledgements: NIH/NIGMS/R16 (1R16GM145556-01A1)