Nanoformulation for anticancer drug delivery: Enhanced pharmacokinetics and circulation.

摘要

In this study, we have explored the application of the Layer-by-Layer (LbL) assembly technique for improving injectable drug delivery systems of low soluble anticancer drugs (e.g. Camptothecin (CPT), Paclitaxel (PTX) or Doxorubicin (DOX)). For this study, a polyelectrolyte shell encapsulates different types of drug nanocores (e.g. soft core, nanomicelle or solid lipid nanocores).The low soluble drugs tend to crystallize and precipitate in an aqueous medium. This is the reason they cannot be injected and may have low concentrations and low circulation time in the blood. Even though these drugs when present in the cancer microenvironment have high anti-tumor inhibition, the delivery to the tumor site after intravenous administration is a challenge. We have used FDA-approved biopolymers for the process and elaborated formation of 60-90 nm diameter initial cores, which was stabilized by multilayer LbL shells for controlled release and longer circulation. A washless LbL assembly process was applied as an essential advancement in nano-assembly technology using low density nanocore (lipids) and preventing aggregation. This advancement reduced the number of process steps, enhanced drug loading capacity, and prevented the loss of expensive polyelectrolytes.;Finally, we elaborated a general nano-encapsulation process, which allowed these three important anticancer drug core-shell nanocapsules with diameters of ca. 100-130 nm (this small size is a record for LbL encapsulation technique) to be stable in the serum and the blood for at least one week, efficient for cancer cell culture studies, injectable to mice with circulation for 4 hrs, and effective in suppressing tumors. This work is divided into three studies. The first study (CHAPTER 4) explores the application of LbL assembly for encapsulating a soft core of albumin protein and CPT anticancer drug. In order to preserve the activity of drug in the core, a unique technique of pH reversal is employed where the first few layers of the LbL shell are assembled at acidic pH 3, and the final layers (2-3) are assembled at a slightly basic pH of 7.4. These LbL-encapsulated nanocores are not stable and immediately aggregate in water or the serum. A final layer of 5 kDa PEG was assembled to improve circulation time. It showed higher colloidal stability in PBS, high drug loading concentration of 0.5 mg/mL, and an improved drug chemical stability in Fetal Bovine Serum with high lactone fraction of 99%. It also showed 3 times improved cytotoxicity against glioblastoma cancer cells. For the first time we applied a new method of the LbL capsule assembly at different pH values, the first 4 bilayers at pH 3, and the following 3 bilayers at pH 7.4.;In the second study (CHAPTER 5), the developed LbL assembly for low solubility drug encapsulation was extended for the delivery of PTX loaded in nanomicelle cores. PTX, as a nanomicelle core, is encapsulated with fewer layers of LbL assembly, followed by an extra layer of PEG (PEGylation). A significant improvement was seen in reducing the process steps through reduction in the number of LbL layers, while smaller nano-colloids, ~100 nm, were produced with improved drug loading capacity, higher cytotoxicity, and high mice survival rate.;In the third study (CHAPTER 6), we have applied the concepts learned and the techniques developed from the previous two studies to modify the surface of the nanostructured solid lipid carriers (NLC) with LbL architecture, plus extra PEGylation. The NLC are co-loaded with DOX and docosahexaenoic acid (DHA). This study is an attempt to further increase drug circulation time in the blood. We improved the colloidal stability with a narrow distribution size, 128 nm, polydispersity of 0.098, a higher longevity in the blood, a 1.5 times lower accumulation in the liver, a 2.25 times higher accumulation in tumors, and a significant ~3.5 times greater tumor growth inhibition in 4T1 murine tumor model in mice.;In conclusion, we developed a general model of an LbL nanoassembly core-shell drug delivery system of three anticancer drugs. The capsules had diameters of ca. 100170 nm, were stable in the serum and the blood for three weeks, were injectable to small animals with a circulation time of 1-4 hrs., and effectively suppressed cancerous tumors in mice.