Institute for BioNanotechnology in Medicine ROMP-Based Polymer Nanoparticles for The Targeted Delivery of Antitumor Agents Paul A. Bertin, DeeDee Smith, Julianne M. Gibbs, Sang-Min Lee, Courtney Phillips, Emily Pentzer, and SonBinh T. Nguyen Department of Chemistry and Center for Nanofabrication and Molecular Self-Assembly Northwestern University, Evanston, IL 60208 Institute for BioNanotechnology in Medicine Julianne M. Gibbs Paul A. Bertin DeeDee Smith Sang-Min Lee Institute for BioNanotechnology in Medicine Nanotechnology and Medicine Chemistry Physics Engineering Bio- Nanotechnology Biology Medicine Opportunities: • drug delivery • diagnostics • imaging Challenges: • design insight • Multifunctional, multiscale synthesis • disease detection • improve therapy Nanostructured Tools Inspiration “Nanotechnology has the potential to make significant contributions to disease prevention, detection, diagnosis, and treatment.” ** ** Srinivas, P. R. Laboratory Investigation 2002, 82, 657-662. Institute for BioNanotechnology in Medicine Hanahan, D.; Weinberg, R. A. Cell 2000, 100, 57-70. * American Cancer Society: www.cancer.org Self-sufficiency in growth signals Insensitive to anti-growth signals Evading apoptosis Sustained angiogenesis Tissue invasion & Metastasis Infinite replicative potential Selectivity of existing cytotoxic chemotherapy for malignant cells over healthy cells is low, resulting in life-threatening side effects. How can selectivity be improved?? Cancer: An Attractive Target 2005 Projection: 1.37 million new cases in US alone; 570,000 deaths * Institute for BioNanotechnology in Medicine Motivation for Targeted Drug Delivery Research • Problems associated with small molecule drugs include: – Poor water solubility – Serious side effects from interaction with non-target cells – Rapidly eliminated from the body – Drug metabolism decreases activity and increases toxicity • Research efforts focus on the design of appropriate “bullets” to address these problems  Soluble polymers  Microcapsules  Liposomes  Polymeric Micelles and Nanoparticles Why do we need a “magic bullet”? Institute for BioNanotechnology in Medicine 1. Core-shell structure 3. Spatial Recognition 2. Biocompatible 4. Temporal Structure Adenovirus Type 2 Virus—A Natural Delivery Vehicle Institute for BioNanotechnology in Medicine Polymers as Functional Materials F F F F F F F F • Capitalize upon poly(F) as ‘amplified monomers’. Inorganic & Surface Science Organic & Polymer Chemistry Biological Chemistry Institute for BioNanotechnology in Medicine Advantages of Polymer Therapeutics • Increased solubility/stability • Reduced immunogenicity • Prolonged plasma half-lie through prevention and avoidance of RES and MPS • Requires less frequent dosing “Nanotechnology applied to medicine will bring significant advancement to the treatment of cancer. These hybrid systems can be viewed as the 1st generation of nanomedicines.” “Using advanced polymer chemistry and precision engineering at the molecular level, together with an appreciation of the pathophysiology of normal and disease tissue will help to realize the full therapeutic potential of the post-genomics era.” Duncan et al. TRENDS in Biotechnology 2006, 24, 39-47. Institute for BioNanotechnology in Medicine Passive Targeting Systems PEO-b-p(Asp-Hyd-DOX) Kataoka, K. et al. Angew. Chem. Int. Ed. 2003, 42, 4640-4643. O NH OH O NH NH O HN O Ph O NH N H O O HO O HO O HO OH HO O O O 95 5 Duncan, R. et al. J. Controlled Release 1992, 19, 331-346. HPMA Peptide Spacer Doxorubicin (DOX) DOX Hydrophobic Hydrophilic 65 nm combretastatin Sengupta, S. et al. Nature 2005, 436, 568-572. DOX-PLGA PEG-liposome “Nanocell” 200 nm PK1 Institute for BioNanotechnology in Medicine Requirements for Drug Delivery • Drug delivery particle should self-assemble from drug molecules, components for targeting, and components for stability • Particle size should enable effective penetration in target tissue and cells (< 400 nm for tumor tissue) • Drug delivery particles should be stable and biologically inert before reaching their target • Drug release should result from interactions with the target cells • Components of drug delivery system should easily be removed from the organism [...]... Equilibrium favors polymer formation due to the alleviation of ring strain • Highly functional group tolerant, works at mild temperature • Amenable to the synthesis of monodisperse block copolymers Institute for BioNanotechnology in Medicine A Versatile ROMP Synthon: 100% exo-Norbornen-2-ol X OH Aldrich - 55% endo + H X X X Diels-Alder - >85% endo O OH KOH O O Keith J Watson OH 100% exo Institute for BioNanotechnology... BioNanotechnology in Medicine A Toolbox for Monomer Development I O O O OH Cl n O OH O Br O >120 new monomers for ROMP Watson et al Institute for BioNanotechnology in Medicine ROMP- Based Nanostructures amphiphilic block copolymers Cl drug Cl drug PCy3 Ru CHPh PCy3 hydrophobic hydrophilic biocompatible group self-assembly ≤ 400 nm Institute for BioNanotechnology in Medicine Block Copolymer Synthesis O Ru* Ph HN n 1... into a biodegradable or removable (< 45 kDa) platform • Hydrophilic components should resist protein adsorption and prevent secondary aggregation • Targeting groups on the surface should allow for site-specific drug delivery Ferrari, M Nature Rev Cancer 2005, 5, 161-171 Institute for BioNanotechnology in Medicine Ring-Opening Metathesis Polymerization (ROMP) M n + Y M X n X Y PCy3 Cl Ru Cl m CHPh PCy3... 3793-3795 in Medicine Polymer Synthesis PEO15-b-DOX15 PEO15 Polymer Mw (Daltons) PDI PEO15 Theoretical Mw of PEO15-b-DOX15 = 17901 Mn (Daltons) 6300 10236 1.19 PEO15-bDOX15 16259 17949 1.12 Bertin, P., Smith, D., Nguyen, S T Chem Comm 2005, 3793-3795 Institute for BioNanotechnology in Medicine Nanoparticle Formation Particles show narrow size distribution around 230 nm Institute for BioNanotechnology... peptide, sugar, antibody Tumor Tissue Ligand -Targeted Therapeutic • Many cancer cells overexpress receptors related to signaling pathways for cell growth (HER-2, ανβ3 integrin, folate receptor) • Ligand -targeted therapeutics allow for selective tumor uptake Institute for BioNanotechnology in Medicine Surface-Modifiable Particles 10 3 119 14 2 Bertin and Davis Institute for BioNanotechnology in Medicine ... DIC+DOX DOX With Clifton Shen Institute for BioNanotechnology in Medicine Passive Uptake 63X DAPI+DOX (fixed) With Clifton Shen Institute for BioNanotechnology in Medicine Neuroblastoma Medline Plus: www.nlm.nih.gov/medlineplus/neuroblastoma.html Cancer affecting immature nerve cells, most commonly in the adrenal glands and the sympathetic nervous system ganglia of the abdomen Common drugs used to treat... sores, depression of immune system, bone marrow suppression Most common cancer in infants with 650 new cases each year In 7 of 10 cases the disease is not diagnosed until it has metathesized Institute for BioNanotechnology in Medicine Free Doxorubicin vs Doxorubicin PNP SKN-SH Wild Type Cells SKN-SH Wild Cells With Dr Bernard Mirkin SKN-SH Wild Cells with Dox Nanoparticles Institute for BioNanotechnology... Resistant Cells with DOX Nanoparticles Institute for BioNanotechnology in Medicine Cholesterol Control Institute for BioNanotechnology in Medicine Cholesterol Control Polymers polymer Mn (Daltons) Mw (Daltons) PDI Theoretical Mn PEO15-b-Chol15 13807 16900 1.22 16690 PEO6-b-Chol30 21551 22234 1.03 21096 PEO9-b-Chol28 18980 19509 1.02 21424 PEO12-b-Chol25 21798 22862 1.04 21153 Institute for BioNanotechnology...Polymeric Nanoparticles as Model Drug Delivery Vehicles Institute for BioNanotechnology in Medicine Magic Bullet? Advantage: multicomponent payload per biorecognition event • Core-shell nanostructures with control over morphology, size, and size distribution • Incorporate multiple therapeutic agents (drug, DNA, protein), imaging agents, and penetrating peptides into... 37, 8364-8372 Institute for BioNanotechnology in Medicine Evidence of Core-Shell Structure Ph 35 IND 15 1 DMSO 2 Dialysis PEO 166 nm CDCl3 CDCl3 1 Lyophilize 2 D2O Bertin, P A et al Macromolecules 2004, 37, 8364-8372 Bertin Institute for BioNanotechnology in Medicine In Vitro Drug Release Dialysis Experiment pH = 3.0 In vitro release of indomethacin from copolymer IND35-b-PEO7 nanoparticles after 48 . Institute for BioNanotechnology in Medicine ROMP-Based Polymer Nanoparticles for The Targeted Delivery of Antitumor Agents Paul A. Bertin, DeeDee Smith, Julianne. • Components of drug delivery system should easily be removed from the organism Institute for BioNanotechnology in Medicine Polymeric Nanoparticles as Model Drug Delivery Vehicles Institute for BioNanotechnology. Medicine • Equilibrium favors polymer formation due to the alleviation of ring strain • Highly functional group tolerant, works at mild temperature • Amenable to the synthesis of monodisperse block copolymers Ring-Opening