Transepithelial transport of nanoparticles targeted to the neonatal Fc receptor for oral delivery applications

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering, 2013.Cataloged from PDF version of thesis.Includes bibliographical references.Nanoparticles (NPs) are poised to have a tremendous impact on the treatment of many diseases, but their broad application is limited because currently they can only be administered by parenteral methods. Oral administration of NPs is highly preferred because of the convenience and compliance by patients, but remains a significant challenge because of the barriers presented by the gastrointestinal tract. In particular, transport across the intestinal epithelium limits efficient oral delivery of NPs. The neonatal Fc receptor (FcRn) mediates IgG antibody transport across epithelial barriers. It was discovered as the receptor in the neonatal intestine that transports IgG in breast milk from mother to offspring. However, FcRn is expressed into adulthood at levels similar to fetal expression. FcRn interacts with the Fc portion of IgG in a pH-dependent manner, binding with high affinity at acidic (<6.5) but not neutral pH (7.4). Targeting NPs to FcRn using IgG Fc fragments was hypothesized to enable orally administered NPs to be transported across the intestinal epithelium. FcRn-targeted NPs were formulated using poly(lactic acid)-b-polyethylene glycol (PLA-PEG) block copolymers and engineered to have particle sizes less than 100 nm with IgG Fc conjugated to the surface. Transepithelial transport of the NPs was first evaluated in an in vitro cell monolayer transport model using Caco-2 cells. FcRn-targeted NPs were transported across the monolayer at a rate twice that of non-targeted NPs. The transport rate was reduced significantly when excess IgG was added along with the FcRn-targeted NPs. Next, FcRn-targeted NPs were then evaluated using in vivo mouse models. Fluorescent FcRn-targeted NPs were observed with fluorescent microscopy crossing the intestinal epithelium and entering the lamina propria after oral administration. Using radiolabeled NPs, orally administered FcRn-targeted NPs were detected in the liver, lungs, and spleen with a mean absorption efficiency of 13.7% for FcRn-targeted NPs compared with only 1.2% for non-targeted NPs. Finally, insulin was encapsulated in the NPs to evaluate the FcRn-targeted NPs as a NPbased therapeutic. In wild-type mice, orally administered FcRn-targeted NPs containing insulin were able to generate a prolonged hypoglycemic response using a clinically relevant insulin dose of 1.1 U/kg. The response was specifically due to FcRn, as studies in FcRn knockout mice mitigated the enhanced response of the FcRn-targeted NPs. This technology has the potential to have an impact on the treatment of many diseases by enabling NP-based therapies to be administered orally. In addition, the encapsulation of drugs or biologics that are currently limited by low bioavailability into FcRn-targeted NPs may enable markedly more efficient oral delivery of the therapies.by Eric M. Pridgen.Ph.D

Synergistic cytotoxicity of irinotecan and cisplatin in dual-drug targeted polymeric nanoparticles

Aim: Two unexplored aspects for irinotecan and cisplatin (I&C) combination chemotherapy are: actively targeting both drugs to a specific diseased cell type, and delivering both drugs on the same vehicle to ensure their synchronized entry into the cell at a well-defined ratio. In this work, the authors report the use of targeted polymeric nanoparticles (NPs) to coencapsulate and deliver I&C to cancer cells expressing the prostate-specific membrane antigen. Materials & methods: Targeted NPs were prepared in a single step by mixing four different precursors inside microfluidic devices. Results: I&C were encapsulated in 55-nm NPs and showed an eightfold increase in internalization by prostate-specific membrane antigen-expressing LNCaP cells compared with nontargeted NPs. NPs coencapsulating both drugs exhibited strong synergism in LNCaP cells with a combination index of 0.2. Conclusion: The strategy of coencapsulating both I&C in a single NP targeted to a specific cell type could potentially be used to treat different types of cancer.Prostate Cancer Foundation (Nanotherapeutics Award)MIT-Harvard Center of Cancer Nanotechnology Excellence (U54-CA151884)National Science Foundation (U.S.). Graduate Research Fellowship ProgramAmerican Society for Engineering Education. National Defense Science and Engineering Graduate Fellowshi

DNA Self-Assembly of Targeted Near-Infrared-Responsive Gold Nanoparticles for Cancer Thermo-Chemotherapy

Targeted Cancer Therapy: Inspired by the ability of DNA hybridization, a targeted near-infrared (NIR) light-responsive delivery system has been developed through simple DNA self-assembly (PEG=polyethylene glycol). This DNA-based platform shows the ability of releasing therapeutics upon near-infrared irradiation, and remarkable targeted thermo- and chemotherapeutic efficacy in vitro and in vivo

Microfluidic Platform for Combinatorial Synthesis and Optimization of Targeted Nanoparticles for Cancer Therapy

Taking a nanoparticle (NP) from discovery to clinical translation has been slow compared to small molecules, in part by the lack of systems that enable their precise engineering and rapid optimization. In this work we have developed a microfluidic platform for the rapid, combinatorial synthesis and optimization of NPs. The system takes in a number of NP precursors from which a library of NPs with varying size, surface charge, target ligand density, and drug load is produced in a reproducible manner. We rapidly synthesized 45 different formulations of poly(lactic-co-glycolic acid)-b-poly(ethylene glycol) NPs of different size and surface composition and screened and ranked the NPs for their ability to evade macrophage uptake in vitro. Comparison of the results to pharmacokinetic studies in vivo in mice revealed a correlation between in vitro screen and in vivo behavior. Next, we selected NP synthesis parameters that resulted in longer blood half-life and used the microfluidic platform to synthesize targeted NPs with varying targeting ligand density (using a model targeting ligand against cancer cells). We screened NPs in vitro against prostate cancer cells as well as macrophages, identifying one formulation that exhibited high uptake by cancer cells yet similar macrophage uptake compared to nontargeted NPs. In vivo, the selected targeted NPs showed a 3.5-fold increase in tumor accumulation in mice compared to nontargeted NPs. The developed microfluidic platform in this work represents a tool that could potentially accelerate the discovery and clinical translation of NPs.Prostate Cancer Foundation (Award in Nanotherapeutics)National Cancer Institute (U.S.) (Center of Cancer Nanotechnology Excellence at MIT-Harvard U54-CA151884National Heart, Lung, and Blood Institute (Programs of Excellence in Nanotechnology HHSN268201000045C)National Science Foundation (U.S.). Graduate Research FellowshipAmerican Society for Engineering Education. National Defense Science and Engineering Graduate FellowshipNational Cancer Institute (U.S.) (Center of Cancer Nanotechnology Excellence. Graduate Research Fellowship

Polymeric nanoparticle drug delivery technologies for oral delivery applications

Introduction: Many therapeutics are limited to parenteral administration. Oral administration is a desirable alternative because of the convenience and increased compliance by patients, especially for chronic diseases that require frequent administration. Polymeric nanoparticles are one technology being developed to enable clinically feasible oral delivery. Areas Covered: This review discusses the challenges associated with oral delivery. Strategies used to overcome gastrointestinal barriers using polymeric nanoparticles will be considered, including mucoadhesive biomaterials and targeting of nanoparticles to transcytosis pathways associated with M cells and enterocytes. Applications of oral delivery technologies will also be discussed, such as oral chemotherapies, oral insulin, treatment of inflammatory bowel disease, and mucosal vaccinations. Expert Opinion: There have been many approaches used to overcome the transport barriers presented by the gastrointestinal tract, but most have been limited by low bioavailability. Recent strategies targeting nanoparticles to transcytosis pathways present in the intestines have demonstrated that it is feasible to efficiently transport both therapeutics and nanoparticles across the intestines and into systemic circulation after oral administration. Further understanding of the physiology and pathophysiology of the intestines could lead to additional improvements in oral polymeric nanoparticle technologies and enable the translation of these technologies to clinical practice

Transepithelial Transport of Fc-Targeted Nanoparticles by the Neonatal Fc Receptor for Oral Delivery

Nanoparticles are poised to have a tremendous impact on the treatment of many diseases, but their broad application is limited because currently they can only be administered by parenteral methods. Oral administration of nanoparticles is preferred but remains a challenge because transport across the intestinal epithelium is limited. We show that nanoparticles targeted to the neonatal Fc receptor (FcRn), which mediates the transport of immunoglobulin G antibodies across epithelial barriers, are efficiently transported across the intestinal epithelium using both in vitro and in vivo models. In mice, orally administered FcRn-targeted nanoparticles crossed the intestinal epithelium and reached systemic circulation with a mean absorption efficiency of 13.7%*hour compared with only 1.2%*hour for nontargeted nanoparticles. In addition, targeted nanoparticles containing insulin as a model nanoparticle-based therapy for diabetes were orally administered at a clinically relevant insulin dose of 1.1 U/kg and elicited a prolonged hypoglycemic response in wild-type mice. This effect was abolished in FcRn knockout mice, indicating that the enhanced nanoparticle transport was specifically due to FcRn. FcRn-targeted nanoparticles may have a major impact on the treatment of many diseases by enabling drugs currently limited by low bioavailability to be efficiently delivered though oral administration.Prostate Cancer Foundation (Award in Nanotherapeutics)National Cancer Institute (U.S.) (Center for Cancer Nanotechnology Excellence U54-CA151884)National Heart, Lung, and Blood Institute (Program of Excellence in Nanotechnology Award Contract HHSN268201000045C)National Institutes of Health (U.S.) (Grant EB000244)National Institutes of Health (U.S.) (R01 Grant EB015419-01)American Society for Engineering Education. National Defense Science and Engineering Graduate FellowshipNational Cancer Institute (U.S.) (Center for Cancer Nanotechnology Excellence Graduate Research Fellowship 5 U54 CA151884-02

Synergistic cytotoxicity of irinotecan and cisplatin in dual-drug targeted polymeric nanoparticles

Aim: Two unexplored aspects for irinotecan and cisplatin (I&C) combination chemotherapy are (1) actively targeting both drugs to a specific diseased cell type and (2) delivering both drugs on the same vehicle to ensure their synchronized entry into the cell at a well-defined ratio. In this work we report the use of targeted polymeric nanoparticles (NPs) to co-encapsulate and deliver I&C to cancer cells expressing the Prostate Specific Membrane Antigen (PSMA). Method: We prepared targeted NPs in a single-step by mixing four different precursors inside microfluidic devices. Results: I&C were encapsulated in 55-nm NPs and showed an 8-fold increase in internalization by PSMA-expressing LNCaP cells compared to non-targeted NPs. NPs co-encapsulating both drugs exhibited strong synergism in LNCaP cells with a combination index of 0.2. Conclusion: The strategy of co-encapsulating both irinotecan and cisplatin in a single NP targeted to a specific cell type could potentially be used to treat different types of cancer

DNA Self-Assembly of Targeted Near-Infrared-Responsive Gold Nanoparticles for Cancer Thermo-Chemotherapy

Targeted cancer therapy: Inspired by the ability of DNA hybridization, a targeted near-infrared (NIR) light-responsive delivery system has been developed through simple DNA self-assembly (see picture; PEG=polyethylene glycol). This DNA-based platform shows the ability of releasing therapeutics upon near-infrared irradiation, and remarkable targeted thermo- and chemotherapeutic efficacy in vitro and in vivo.National Institutes of Health (U.S.) (Grant CA151884)Prostate Cancer Foundation (Program in Cancer Nanotherapeutics

Nanoparticle Encapsulation of Mitaplatin and the Effect Thereof on In Vivo properties

Nanoparticle (NP) therapeutics have the potential to significantly alter the in vivo biological properties of the pharmaceutically active agents that they carry. Here we describe the development of a polymeric NP, termed M-NP, comprising poly(d,l-lactic-co-glycolic acid)-block-poly(ethylene glycol) (PLGA-PEG), stabilized with poly(vinyl alcohol) (PVA), and loaded with a water-soluble platinum(IV) [Pt(IV)] prodrug, mitaplatin. Mitaplatin, c,c,t-[PtCl[subscript 2](NH[subscript 3])[subscript 2](OOCCHCl[subscript 2])[subscript 2]], is a compound designed to release cisplatin, an anticancer drug in widespread clinical use, and the orphan drug dichloroacetate following chemical reduction. An optimized preparation of M-NP by double emulsion and its physical characterization are reported, and the influence of encapsulation on the properties of the platinum agent is evaluated in vivo. Encapsulation increases the circulation time of Pt in the bloodstream of rats. The biodistribution of Pt in mice is also affected by nanoparticle encapsulation, resulting in reduced accumulation in the kidneys. Finally, the efficacy of both free mitaplatin and M-NP, measured by tumor growth inhibition in a mouse xenograft model of triple-negative breast cancer, reveals that controlled release of mitaplatin over time from the nanoparticle treatment produces long-term efficacy comparable to that of free mitaplatin, which might limit toxic side effects.National Institutes of Health (U.S.) (Grant 5-U54-CA119349)National Institutes of Health (U.S.) (Grant 5-U54-CA151884)National Institutes of Health (U.S.) (Grant 5-R01-CA034992)National Institutes of Health (U.S.) (MIT-Harvard Center of Cancer Nanotechnology Excellence. Grant 5-U54-CA151884-02)National Institutes of Health (U.S.) (MIT-Harvard Center of Cancer Nanotechnology Excellence. Grant 5-U54-CA151884)German Academic Exchange Service (Research Fellowship)National Institutes of Health (U.S.) (1-S10-RR13886-01