Nanoformulation of the BDNF for Treatment of Stroke

2015/09/15-2017/07/31        1R21NS088152-01A1   National Institute of Neurological Disorders and Stroke (NINDS)

KABANOV, ALEXANDER (PI)

 

nanobdnf
Nano-BDNF spontaneously forms in water upon simple mixing of the native BDNF and PEG-PGA block copolymer. BDNF electrostatically couples with the oppositely charged PGA chain and entraps into a particle core surrounded by a shell of uncharged water-soluble PEG. Active BDNF is released upon interaction with cell receptor or BBB transporter

This project develops nanoformulation for the delivery of the brain-derived neurotrophic factor (BDNF) to the central nervous system and evaluates the drug candidate using this formulation for the treatment of ischemic stroke. BDNF has shown potential to exert neuroprotective and neuroregenerative effects when administered after stroke. However, the use of native BDNF as therapeutic agent for systemic administration is hindered by its poor brain accumulation. To address this problem we propose a novel yet simple and scalable polymeric nano-formulation of BDNF, nano-BDNF, in which BDNF is incorporated into polyion complexes with safe and biocompatible poly(ethylene glycol)-poly(L-glutamate) (PEG-PGA) block copolymer. The complexes are produced spontaneously in mild aqueous conditions upon mixing of native BDNF with the PEG-PGA block copolymer, which entraps the BDNF molecule in nanoscale size (<100 nm) core-shell particles. Preliminary studies suggest that nano-BDNF will strongly improve the brain uptake of BDNF and increase efficacy of BDNF treatment to elicit neuroprotection and neuro-restoration after stroke. We propose to advance nano-BDNF as a potential therapeutic agent for the treatment of brain injury in an animal model of stroke. This will be achieved by (1) producing in house the abundant amount of human recombinant BDNF and developing simple, scalable and robust procedure for nano-BDNF with optimized composition; (2) determining the brain pharmacokinetics of nano-BDNF after intravenous administration and identifying the best composition for efficacy studies; and (3) determining the therapeutic effect of nano-BDNF treatment on chronic recovery in an ischemia/reperfusion-induced mouse middle cerebral artery occlusion (MCAO) model. We use step-wise process; each step has its own go/no go criteria to identify a lead nano-BDNF candidate that will be taken forward into preclinical development. The plan for subsequent therapy development will implement the product development steps and result in the filing of IND.

 

Carolina Center for Cancer Nanotechnology excellence: Nano Approaches to Modulate Host Cell Response for Cancer Therapy. Project 4: High Capacity Polymeric Micelle Therapeutics for Lung Cancer

2015/09/15-2020/07/31        1U54CA198999-01       National Cancer Institute (NCI)

HUANG, LEAF (PI), TEPPER, JOEL (CPI), KABANOV, ALEXANDER V (PI, Project 4), WANG, ANDREW (CPI), PECOT, CHAD (CPI)

 

CCNE pr4The central goal of this project is to improve systemic therapies for non-small cell lung cancer (NSCLC) using combinations of potent anticancer agents, chemosensitizers, and agents that target the tumor microenvironment (TME). Many highly promising small-molecule cancer-targeting therapeutics fail due to poor solubility, stability and other delivery related problems. Recent advances in nanoparticle (NP) drug delivery vehicles provide a unique opportunity to “rescue” these agents for clinical application. This is the focal point of our research, which is to use NPs that can incorporate such agents and preferentially deliver them to tumors. Our group has successfully developed a novel drug delivery platform based on poly(2-oxazoline) (POx) polymeric micelles, that is well suited for the delivery of poorly soluble active drugs, such as paxclitaxel (PTX). Compared to conventional formulations and other NP platforms, our POx platform is unique in its high drug loading capacity. Our preliminary data showed that such high drug loading translated into lower toxicity and high therapeutic efficacy when we compared POx/PTX to both Taxol® and Abraxane®. The National Characterization Laboratory has evaluated the PTX formulation using our lead POx block copolymer of poly(2-butyl-2-oxozaline) (BuOx) and poly(2-methyl-2-oxazoline) (MeOx), and concluded that both the copolymer and formulation lack immunological and hematological toxicities. Significantly, we have shown that multiple agents can be co-formulated within the same POx micelles, enabling co-delivery of anticancer agents, chemosensitizers, and TME-modifying compounds. Such combinations can increase cancer cell cytotoxicity and modify the TME to enhance tumor control. We hypothesize that POx micelles can serve as a powerful and versatile platform for delivery of such agents. We assembled a cross-disciplinary team of physician-scientists and experts in nanotechnology, pharmacology, and chemoinformatics, and will focus our research efforts on using POx micelle therapeutics in NSCLC, a disease that despite treatment advances, still has a very poor outcome. The Specific Aims are: 1. Develop a predictive computational model for rapid selection and incorporation of new and existing anticancer agents, chemosensitizers, and TME modifiers into POx micelles. 2. Evaluate chemosensitizers and anticancer agents incorporated in POx micelles as therapeutic modalities to improve treatment of NSCLC. 3. Evaluate polymeric micelle formulations of agents that target the TME as a treatment strategy. The computational model for rational design of formulations will drastically increase the throughput and allow us to further develop new anti-cancer therapeutics through trans-Alliance collaborations and other NCI mechanisms. Successful therapeutics will be advanced for further development.

 

Targeted Core Shell Nanogels for Triple Negative Breast Cancer

2015/08/14-2020/07/31        1U01CA198910-01       National Cancer Institute (NCI)

KABANOV, ALEXANDER V (PI), BRONICH, TATIANA (CPI), LIU, RIHE (CPI)

nanogels u01
(a) The onion-type and (b) the raspberry type CSNGs.

The central goal of this project is to improve systemic therapies of cancer using soft nanomaterials that can deeply penetrate into tumors and deliver potent anticancer agents to targeted cancer cells. Many small-molecule therapeutics that were highly promising for cancer therapy, eventually failed clinical translation due to toxicity, poor solubility, stability and other delivery related problems. Recent advances in nanoparticle drug carriers provide a unique opportunity to “rescue” these agents and enable their clinical application. This is the focal point of our research using nanocarriers that can incorporate such agents, and preferentially deliver them to tumors. Our group has successfully developed a novel platform for drug delivery that uses aqueous polymeric gel nanoparticles, core-shell nanogels (CSNGs). CSNGs are manufactured through a proprietary self-assembly process and can be readily filled with various drug payloads. They are water-swollen and are practically non-adhesive, which may diminish their off-target side effects. We hypothesize that (a) the systemic and tumor flow dynamics of CSNGs will be a function of their molecular architecture and mechanical properties (b) and that these properties can be rationally controlled to modify the PK, distribution and tumor penetration of the CSNGs and the drugs they deliver. We will focus our research efforts on using CSNGs in the triple negative breast cancer (TNBC), a disease that despite treatment advances, still has a very poor outcome. As targeting strategies we will use the novel single-domain polypeptide antagonists of the EGFR and HER3 that are frequently overexpressed in TNBC and are associated with higher risk of mortality in TNBC. We assembled a cross-disciplinary team of physician-scientists and experts in nanotechnology and pharmacology, and will focus our research efforts on using CSNGs-based therapeutics in TNBC, a disease that despite treatment advances has a very poor outcome. The specific aims are: 1. Determine how the molecular architecture and mechanical properties of polypeptide-based CSNGs affects their ability to load, deliver and release therapeutic cargos. 2. Determine how the structure and mechanical properties of the drug-loaded CSNGs affect the in vivo PK and tumor distribution of the drugs and nanogels in murine models of TNBC. 3. Develop EGFR and HER3 targeted drug-loaded CSNGs with maximal tumor penetration, maximal delivery of drug payload to tumors and potent anti-tumor activity in TNBC. These integrative efforts will address major barriers in developing novel nanotechnology platforms for the treatment of TNBC, and facilitate our understanding of the cancer biology and the mechanisms of in vivo delivery. If successful, we will determine a new targeted formulation of chemotherapeutic drugs in CSNGs that may be highly effective in treatment of TNBC. The results will then be reviewed with our clinical advisors for further consideration of these formulations for translational and clinical development.

 

Carolina Cancer Nanotechnology Training Program (C-CNTP)

2015/07/01-2020/06/30        1T32CA196589-01       National Cancer Institute (NCI)

KABANOV, ALEXANDER V (PI), WANG, ANDREW (CPI)

This proposal seeks to establish the Carolina Cancer Nanotechnology Training Program (CCNTP). The goal of the program is to make a major contribution to the growth of the cancer nanotechnology workforce by providing training and research experiences to a highly select cohort of postdoctoral fellows. We have assembled a team of 22 outstanding Program faculty from 11 departments and 3 schools at the University of North Carolina Chapel Hill with specific expertise in physical and material sciences, biomedical engineering, drug delivery, computational modeling as well as basic biomedical research and clinical science, all of whom have demonstrated strong interests, capabilities and collaborations at the interface between nanoscience and cancer. The objectives of the C-CNTP are to: 1) recruit an elite group of talented postdoctoral fellows from diverse backgrounds with PhD or MD and provide them with outstanding postdoctoral experience including focused didactic training and co-mentored research experience with faculty mentors from complementary fields; 2) provide each trainee with Intensive Integrated Learning Accelerating Module training in conjunction with flipped classrooms followed by workshops and didactic courses to remediate differences in their backgrounds and to deepen the knowledge and understanding in the key areas of cancer nanotechnology; and 3) facilitate transition of trainees to independence by providing them with opportunities to a) conduct original cancer nanotechnology research projects; b) apply for the individual cancer nanotechnology Pilot Grants within C-CNTP, and c) acquire written and oral communications skills needed to publish manuscripts, report results, and write successful individual extramural support applications focused on problems of cancer nanotechnology. With the support of the NIH Ruth L. Kirschstein National Research Service Award our institution will establish a world-class postdoctoral training program that will capitalize on the existing strengths in cancer nanotechnology research, consolidate diverse research and education resources across several academic units, and become a significant contributor to addressing the Nation’s research needs in cancer nanotechnology.

 

Preclinical Development of Atr inhibitor VE-822, Delivered Systemically in Nanoparticles, for Medulloblastoma Therapy

7/1/2015 – 6/30/2017               Alex’s Lemonade Stand Foundation

GERSHON, TIMOTHY R (PI), SOKOLSKY, MARINA (CPI)

We will test the hypothesis that targeting the DNA repair protein Atr can enhance therapy for medulloblastoma, the most common malignant pediatric brain tumor. Radiation and chemotherapy, which act by damaging DNA, confer long-term survival for most medulloblastoma patients. Improved treatment is needed, however, because many patients die of recurrence and survivors have life-long neuro-cognitive injury. We hypothesize that medulloblastoma is particularly sensitive to Atr inhibition because prior investigations demonstrated that neural progenitors that give rise to medulloblastoma cannot survive Atr deletion. Our preliminary studies show that conditional deletion of Atr in medulloblastoma-prone SmoM2 mice prevents tumorigenesis. We now propose to investigate the therapeutic potential of Atr inhibitor VE-822, using SmoM2 mice as a preclinical model. We developed a nanoparticle micelle formulation of VE-822 that crosses the blood-brain barrier when administered IP. This novel formulation causes DNA damage and cell death in medulloblastomas in SmoM2 mice without harming the normal brain. Our Specific Aims will determine optimal use of nanoparticle VE-822 and its mechanism of effect.

 

Liposomal Doxorubicin and Pluronic Combination for Cancer Therapy

2015/01/01-2019/12/31        R01CA184088-02         National Cancer Institute (NCI)

KABANOV, ALEXANDER V (PI), ZAMBONI, WILLIAM (CPI)

PEGylated liposomal doxorubicin (Doxil, PLD) is used clinically to treat ovarian cancer (OC) and breast cancer (BC); however, the response rates of PLD treatment in both diseases need to be improved. Based on strong preliminary data, we propose a novel simple strategy to increase the efficacy of PLD treatment by promoting the release of the active ingredient (doxorubicin, (Dox)) from the liposomal particles directly within the tumor matrix, while concurrently sensitizing this tumor to the drug. In this strategy, amphiphilic Pluronic block copolymers (poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide), PEO-PPO-PEO) are intravenously (IV) administered after the PLD treatment, when the concentration of the liposomal drug within the tumor reaches its maximum. We posit, the copolymer then depots into tumors, incorporates into the PLD particles, and promotes the encapsulated drug release, thus increasing drug bioavailability and improving the tumor response. Moreover, a combination of Dox and Pluronic generated within tumors is highly potent in eliminating multidrug resistant (MDR) and tumor-initiating cells (TIC) that can further improve the therapeutic outcomes in cancer. Our objectives are: to determine mechanism by which administration of Pluronic after the PLD increases the anti-tumor activity; provide proof of principle using genetically engineered mouse models (GEMMs) of OC and BC that closely represent biology and microenvironment of solid tumors in patients; and select Pluronic compositions and treatment regimen to maximize the translational and clinical outcomes. The aims will: 1) determine in vitro release kinetics of Dox from PLD and in vivo pharmacokinetics (PK) of Pluronic to select the best Pluroinic composition, doses and schedule; 2) evaluate PK of PLD alone and in combination with the selected Pluronic(s) to determine the amount of Dox released from liposomes in plasma and drug exposure in tumor; 3) evaluate the anti-tumor activity and safety of the proposed treatments; and 4) determine whether administration of Pluronic after PLD results in depletion of TIC, and decreases tumorigenicity and aggressiveness of cancer cells. The proposed combination therapy if successful has high potential for translation to clinical studies, since it is simple, can improve efficacy of clinically available PLD (such as Doxil), and is likely to be safe, since Pluronics were shown to be safe in clinical trials of non-liposomal Dox/Pluronic formulation, SP1049C.

 

Nanoformulated Brain Derived Neurotrophic Factor (BDNF) for the Treatment of Rett Syndrome

2015/01/01-2016/12/01        HeART Award #3112   Rettsyndrome.org

KABANOV, ALEXANDER V (PI)

Our goal is to develop an effective, simple and safe systemic therapeutic modality for therapy of Rett syndrome (RTT) by delivering the Brain derived neurotrophic factor (BDNF) to the central nervous system (CNS). To address a critical challenge in the delivery of BDNF to the brain, we propose a novel delivery system for BDNF composed of safe and biocompatible polymers that 1) entrap the BDNF molecule in a nanoscale size (under 100 nm) complex (nano-BDNF), 2) increase stability of BDNF against degradation, 3) increase BDNF circulation time and 4) release active BDNF allowing its interaction with cellular receptor. We posit that the proposed nano-BDNF formulation will improve the brain uptake of BDNF and increase efficacy of BDNF treatment to ameliorate RTT. The following aims will be addressed: 1) Express and purify human BDNF at the milligram scale and develop simple, scalable and robust procedure to produce nano-BDNF with optimized composition to ensure BDNF stability, activity and overall safety to neuronal cells. 2) Determine the brain pharmacokinetics and in vivo activity of nano-BDNF to demonstrate improved bioavailability and delivery of active BDNF to the brain after intravenous administration. 3). Validate therapeutic effect of the peripherally administered nano-BDNF using hemizygous male Mecp2tm1.1Bird/Y mice to obtain initial pre-clinical proof of concept for further preclinical investigation in female Mecp2tm1.1Bird/+ mice. Altogether this work will provide evidence that nano-formulation of BDNF can be used to deliver active BDNF to the brain and improve treatment of RTT. If successful the nano-BDNF will be further advanced for clinical development of BDNF therapy for RTT patients.

 

delAQUA – Computer-assisted Drug Solubilization Platform

2014/12/01-2015/11/30           4DR11404                   NC TraCS Institute

KABANOV, ALEXANDER V (PI), TROPSHA, ALEX (CPI)

delAQUA is a patent pending platform technology that uses novel biocompatible poly(2-oxazoline) (POx) polymeric micelles to greatly enhance the solubility of many hydrophobic pharmaceutical active ingredients (APIs). Poor solubility of drug candidates is a common problem in the pharmaceutical industry severely restricting the range of these molecules that can be developed into marketed drugs. The solubilization capacity of the new platform is unprecedented, as has been demonstrated across a broad range of APIs. Our goal is to develop a versatile, cheminformatics-aided high throughput and high capacity nano-formulation platform employing POx-based polymeric micelles for rapid screening, selection and successful solubilization of water insoluble or poorly water-soluble APIs. Towards this goal we will determine structure-solubility correlations that allow predictions for novel formulations of partner compounds through utilization of our lead POx block copolymer, MeOx-b-BuOx-b-MeOx. We will achieve our goal by addressing the following specific aims: 1), To rationally select a library of water-insoluble and poorly soluble APIs in POx polymeric micelle based delivery systems and test these compounds experimentally to form a unique dataset for computational model development; 2) Specific Aim 2. To develop a computational model for reliable selection of hydrophobic APIs predicted to have dramatically (no less that 100-fold) improved solubility in POx polymeric micelles. The successful completion of this pilot grant will lay the groundwork for establishing partnerships with companies pursuing improved solubility of their drugs, as well as leveraging future funding for development and commercialization of promising drug candidates previously limited by solubility issues.

 

PRINT Butyrylcholinesterase (BuChE) Delivery

2013/07/01-2017/06/31        HDTRA1-09-14-FRCWMD-BAA            Defense Threat Reduction Agency (DTRA)

DESIMONE, JOSEPH M (PI), KABANOV, ALEXANDER V (CPI), LUFT, JAMES CHRIS (CPI)

Butyrylcholinesterase (BuChE) is a leading candidate to provide broad spectrum protection against exposure to nerve agents, which pose an ever increasing threat to public and military health. BuChE delivery via appropriately designed particulate formulations offers potential for increased circulation times and rapid onset of action following exposure to nerve agents. The project goal is to develop shape-specific nano- and micro-particles containing active BuChE delivered via transdermal (micro-needle) and inhalation routes of administration that are ideal for accessing local and systemic environments. Particles will be fabricated using a cGMP-compliant, roll-to-roll particle molding technique referred to as PRINT (Particle Replication in Non-wetting Templates). Delivery of nano- and micro-particles will be investigated, with novel cross-linking chemistries used to control the release of BuChE over extended periods of time. There is even a potential for exposure-triggered release by pathophysiologic osmotic imbalance (hyperglycemia and acidosis occur secondary to exposure to organophosphate (OP)-derived nerve agents) that will be explored to enable a bolus release of BuChE from appropriately designed PRINT particles. Particulate BuChE formulations have been scarcely explored, and our ability to precisely control particle design characteristics will provide core knowledge regarding how physical and chemical particle parameters determine the fate of BuChE delivered by these routes. These investigations will enable down selection of PRINT particles parameters necessary to achieve the desired BuChE delivery. In tandem with the “plug and play” opportunities afforded by PRINT, investigations into BuChE delivery by these approaches can pave the way for novel prophylactic and treatment applications using additional protein-based therapies.

 

Polypeptide Modification for Enhanced Brain Delivery

2005/04/01-2016/03/31           R01 NS051334-09       National Institute of Neurological Disorders and Stroke (NINDS)

KABANOV, ALEXANDER V (PI), BANKS, WILLIAM (CPI)

This proposal is a competing continuation of the NIH grant RO1 NS051335. The long term goal is to produce polypeptides that could cross the blood-brain barrier (BBB) and so be developed as central nervous system (CNS) therapeutics. The goal is achieved by modification of polypeptides with Pluronic copolymers (poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide)). The studies have focused on leptin, a candidate for the treatment of epidemic obesity. Leptin failure is mostly due to peripheral resistance – a loss of its ability to cross the BBB necessary to reach its receptors in the arcuate nucleus of the hypothalamus. The previous work produced a conjugate of leptin with Pluronic P85 that is centrally active, has a long half-life in blood, is enzymatically resistant, crosses the BBB by a non-saturable mechanism independently of the leptin transporter, and can exert effects on feeding and body weight after peripheral administration. This proposal will: 1) Optimize modifications to produce superior leptin analogs with maximal BBB permeability, improved half-life in blood and high enzymatic resistance in brain and blood; 2) Identify mechanisms underlying enhanced permeability of leptin analogs in BBB; 3) Determine the permeability of modified leptin analogs across the BBB to select the best analogs for activity testing; and 4) determine the biological activity after intravenous injection of the best leptin analogs 3 in leptin sensitive (ob/ob) and leptin resistant (dietary obese) mouse models of obesity. Analogs completing this development will be a) active in brain, b) have favorable pharmacokinetics, c) be able to cross the BBB in obese mice, and d) reverse obesity after iv injection. Such analogs are candidates for clinical trials in human obesity.

 

High Capacity Nanocarriers for Cancer Chemotherapeutics

2010/09/02-2016/07/31           U01 CA151806-05      National Cancer Institute (NCI)

KABANOV, ALEXANDER V (PI)

Innovative drug delivery platform based on biocompatible poly(2-oxazoline)s block copolymers is proposed to improve chemotherapy of cancer. The technology will overcome major therapeutic challenges in breast cancer. This will be realized by tailoring poly(2-oxazoline)s structure to incorporate very large amounts (> 40 wt. %) of insoluble drugs in polymer micelles of narrow size distribution (20-50 nm). These high-capacity micelles can be lyophilized and re-dispersed without loss of the particle size, drug loading or drug activity. The micelles will carry paclitaxel and docetaxel using drastically less amount of excipients relative to current formulations. Hence, excipient-associated toxicities will be decreased. The micelles will be targeted using monoclonal antibodies to Her2-receptors. This strategy is expected to address Herceptin resistance in cancers overexpressing Her2. The proposed aims will 1) optimize poly(2-oxazoline)s structures for solubilization of water-insoluble drugs; 2) determine pharmacokinetics and maximum tolerated doses in vivo; 3) validate anti-tumor activity in mouse models of breast cancer, and 4) develop targeted micelles effective against Herceptin sensitive and resistant tumors. This CNPP joins together pharmaceutical scientists at UNC Eshelman School of Pharmacy, cancer researchers at Lineberger Comprehensive Cancer Center, and polymer chemists at Technische Universität Dresden who will contribute to the Alliance through challenge projects and other trans-alliance activities.