Module 2 Nanomedicine : A Paradigm Shift in Healthcare

1. Module 2 Nanomedicine: A Paradigm Shift in Healthcare

2. Nanomedicine Changing the Healthcare From symptomatic to pre-emptivemedicine Current symptomatic medicine leads to chronic disease Clinical Symptoms Symptoms Early Diagnosis Preventive Treatment Screening Time Functional and quantitative imaging localizing the disease Minimally invasive treatment Follow the disease response to therapy management tailored to the patient

3. How Conventional Medicine Works for a Disease • Identification of a “diseased state” by a patient who doesn’t “feel right” and then goes or is taken to a clinic • Simple measurements • Follow-up clinical tests • Functional imaging (e.g. x-rays, CAT scans, MRI scans; PET) • Occasional “molecular” tests (gene relocations, amplified gene copies, etc.) to establish genetically determined diseases • Comparison of individual results with “normal ranges” of “normal” individuals. • Surgical repair of injuries (“reconstructive surgery” or removal or diseased tissues or organs • Chemical drugs delivered locally (e.g. ointments to skins, injection of drugs into tissues or organs, etc.) • Chemical drugs delivered systemically (e.g. chemotherapy, etc.) • Stabilization of the patient so that the patient can repair himself/herself https://nanohub.org/resources/3095/about

4. Issues with conventional medicine • Waiting for a patient to feel symptoms means • Disease detection is not early • By the time symptoms are felt, tissue and/or organ destruction has already begin and may be irreversible • Many diseases have similar symptoms making diagnoses based on symptoms at best a guessing game • Cost: Trained people and modern drugs • Diagnostic technologies, if available, are still relatively primitive; or if expensive, are not readily available • Drugs and other treatments are either completely or only crudely targeted to the diseased cells leading to extensive damage to normal bystander cells http://www.bniembarcadero.com/wp-content/uploads/wallet_hospital_400w.jpg

5. Nanomedicine: A Paradigm Shift in Healthcare From symptomatic to pre-emptivemedicine Current symptomatic medicine leads to chronic disease Clinical Symptoms Symptoms Early Diagnosis Preventive Treatment Screening • Challenge? • Understanding root causes of disease • Linking symptoms to molecular origins • Bring bench side science to clinics Time Pan et. al. Eur J Radiol. 2009, 70(2):274-85. Pan et. al. Wiley Interdiscip Rev NanomedNanobiotechnol. 2010 Sep 21. Image source: wiki

6. The ‘Holy Grail’ in Nanomedicine Research What are we really after? • A suitable marker and a high affinity ligand • Preferably peptides, peptidomimmetics) • Affinity: nano molar Can we really package everything in a sensitive, efficacious yet “safer” way? 1. Finding the target 2. Detect early 3. Quantify the disease site 4. Release the drug 5. Follow the drug response 6. Perform all of the above with a “safer” cargo • Sensitive imaging modality • An imaging technique that can provide quantitative information • Deliver therapeutics affecting the normal cells little or none • A carrier that is extremely robust • “Soft”- preferably self-assemblies of polymer or lipids

7. The World of “Nano” A nanometre (Greek: νάνος, nanos, "dwarf"; μέτρον, metrοn) is a unit of length in the metric system, equal to one billionth of a meter (i.e., 10-9 m or one millionth of a millimeter). • Real world scenario: • Humans are 10,000,000 times smaller than the earth. • A 100 nm sized particle, is 10,000,000 times smaller than a human. Scaling down • High surface area • High payload • Multi-functionality • Low overall therapeutics/NP At scales on the order of 100’s of nm, novel materials properties emerge, enabling the development of new class of materials. It can create opportunities for paradigm shifting results, creating new preventive, diagnostic and therapeutic approaches to cancer 100 nm http://www.azonano.com/details.asp?ArticleID=1780

8. Nanomedicine: A Blast from Past? How old is nanotechnology in human history? Lead sulfide crystals (5 nm) Silver nano-colloids were used by Persians, Babylonian and Greek civilizations as antiobiotics Blonde hair The astounding qualities of “nano”-gold were understood by the ancients, who devoted massive amounts of time and energy to alchemy and labeled a primitive form of Nanogold the “Elixir of Life.” It's over 4000 years old. Goes back in ancient Egyptian and Persian times

9. Engineered Nanoparticles for Theranostic Application Homing Ligands Self-assembled polymeric architecture THERAPEUTICS Phospholipids TARGETING CONTRAST 20-200 nm Fibrin: Monoclonal antibody (NIB5F3), peptide Angiogenesis (Integrin):Ab, peptide, peptidomimmetics MRI: Mn, Cu, Fe Optical/Photoacoustics Imaging (PAT): Au, Cu Computed tomography (CT) and Spectral CT: I, Bi, Au, Yb Other functionalization Drugs or Prodrugs for Controlled Delivery Anticancer agents; Radiosensitizers; Radioprotectors; Fibrinolytic agents etc. High payload (40 wt% to 70 wt%) Application areas: Cancer Cardiovascular inflammatory Pan D. 2013 Molecular Pharmaceutics, March, Editorial , Special Issue

10. Basic Concept of Building a Nano-device End product: A Multi-component, Multi-functional “smart” Nanoparticulate system to visualize, characterize and measure biological processes in living systems Anticancer drugs Anti inflammatory Radioisotopes DNA, genes Therapeuticagent • PET • SPECT • Optical • Ultrasound • MRI • Computed Tomography • Photoacoustic Tomography Core materials Gold Metal oxides Metal sulphide Silica Contrast materials PTD Polyarginine Carrier peptide • Antibodies • Proteins • Peptides • Small molecule Homing agent Linker Flexibility Hydrophilicity Charge Length Shell/ coating • Phospholipids • Polyethyleneglycol (PEG) • Carbohydrate • Amphiphilic polymers

11. The Importance of “Bionics”: Invention Inspired by Nature Bionics is the application of biological methods and systems found in nature to the study and design of engineering systems and modern technology. “Biommicry (or Biomimetics) is a new science that studies nature’s models and then imitates or takes inspiration from these designs and processes to solve human problems.” …… Janine M. Benyus “Biomimicry Innovation Inspired by Nature” UC Berkley Collage illustrates gecko adhesion, from toes to nanostructures Adhesive that is the first to master the easy attach and easy release of the reptile's padded feet http://lclark.edu/~autumn

12. Examples of Nano-platforms Gold NanoCages Younan Xia, WUSTL Dendrimer Nano Necklace Nano Wire JC Charlier, Belgium Nano Car (James Tour, Rice U) NanoGold Nanoprobes, Inc. Liposomes Virus NP M Manchester, MG Finn Scripps Research Nano Vault Sarah H. Tolbert , UCLA NanoDots (Quantum Dots) NanoTube Iron Oxide NP FeCo MWNT Nanobox David Gracias, JHU Perfluorocarbon Wickline, Lanza CTRAIN/Kereos, Inc Golden Carbon nanotube Jin-Woo Kim, U Arkansa Nano Bialys Pan (CTRAIN, Washu) “soft” Metal Nanocolloids Pan (CTRAIN, Washu)

13. Size Scales of Nanotechnology “Nano-Bialys” Liposomes Polymeric Micelles 20-60 nm 120-260 nm 100-400 nm Size Vascularly Constrained

14. Understanding the Properties Before Using In Vivo NCL assay cascade Physical Characterization: – Size – Size distribution – Molecular weight – Morphology – Surface area – Porosity – Solubility – Surface charge density – Purity – Sterility – Surface chemistry – Stability – No of ligands, CAs, drugs In Vivo: – Absorption – Pharmacokinetics – Serum half-life – Tissue distribution – Excretion – Safety Plasma PK profile/ Tissue distribution (Liver, lungs, kidney, heart, spleen) In Vitro: – Binding – Pharmacology – Blood contact properties – Cellular uptake – Cytotoxicity

15. The Importance of Size in Distributive Properties of Probes

16. Size Dictates in vivo Distribution of Nanoparticles in the Lymph Nodes! Sentinel lymph node (SLN) imaging is highly relevant in the context of breast cancer staging and may replace fine needle biopsy (FNB) http://www.surgicalprobe.com/ NON INVASIVE IMAGING Real Time IMAGING? MORE GOLD Biomaterials 2010; 31 (14): 4088-4093.

17. Realizing the Importance of Particle Diameter for In Vivo Imaging lGNBs lGNBs c b D=160 nm 20 min 5 min Poor detection D=90 nm a Ctrl Markedly enhanced detection 0.8 0.6 0.4 0.2 D=290 nm NO detection GNB Control GNB GNB Deep lymph node detection may require improved detection sensitivity for robust application and 90nm GNB will markedly enhance efficacy Control GNB Biomaterials 2010; 31 (14): 4088-4093.

18. Targeting Approaches DRUG DRUG DRUG There is a search for dual-mode probes that can detect a tumor imaging) and destroy it (therapy) PROBE PROBE Redox-potential Ultrasound PROBE Temperature pH-sensitive Physical Targeting Active Targeting Passive Targeting Based on nanoparticle functionalization for specific targeting of disease cells Based on retention effect of particle of certain hydrodynamic size in cancerous tissues (e.g. Doxil) Proteins (antibodies and their fragments such as TAT), nucleic acids (aptamers), receptor ligands (peptides, vitamins, and carbohydrates

19. EPR: Taking advantage of retention A. Tumorous tissues suffer of Enhanced Permeability and Retention effect (RES) B. Nanoparticles injected in the blood stream do not permeate through healthy tissues C. Blood vessels in the surrounding of tumorous tissues are defective and porous D. Nanoparticles injected in the blood permeate through blood vessels toward tumorous tissues, wherein they accumulate Annu. Rev. Biomed. Eng. 2007. Vol. 9, pp. 257–88

20. Clinical Example of EPR Doxil is a polyethylene glycol coated liposomal formulation of doxorubicin. Marketed by Ben Venue Laboratories  of J&J. Outside the US, Doxil is known as Caelyx (Janssen). Approved by the FDA for treatment of ovarian cancer and multiple myeloma and an AIDS-related cancer.

21. How Doxil works? Doxorubicin interacts with DNA by intercalation and inhibition of macromolecular biosynthesis. This inhibits the progression of the enzyme topoisomerase II, which relaxes supercoils in DNA for transcription. Doxorubicin stabilizes the topoisomerase II complex after it has broken the DNA chain for replication, preventing the DNA double helix from being resealed and thereby stopping the process of replication.

22. Doxil Side Effects Liposome-encapsulated doxorubicin is less cardiotoxic than free doxorubicin. Polyethylene glycol results in preferential concentration of Doxil in the skin-----a side effect known as hand-foot syndrome. Small amounts of the Dox can leak from capillaries in the palms of the hands and soles of the feet. >50% of patients treated with Doxil developed hand-foot syndrome. Myocet is a non-pegylated liposomal doxorubicin (Phase III) Plenty of room for improvement!

24. Example of an Approved Anticancer Agent Protein-bound paclitaxel is an injectable formulation of paclitaxel, a mitotic inhibitor drug used in the treatment of breast cancer, lung cancer and pancreatic cancer. Paclitaxel Albumin

25. ABRAXANE Clinical Trial Results recTLRR (PRIMARY END POINTS) VS. PACLITAXEL INJECTION FOR ALL RANDOMIZED PATIENTS IN THE PHASE III TRIAL IN METASTATIC BREAST CANCER SIGNIFICANTLY SUPERIOR TUMOR RESPONSE RATE IN ALL RANDOMIZED PATIENTS EFFICACY DEMONSTRATED IN SECOND-LINE METASTATIC PATIENTS AND PATIENTS WHO RELAPSED WITHIN 6 MONTHS OF ADJUVANT CHEMOTHERAPY

26. Is Abraxane Safe? • On Jan, 25, 2014: 1,893 people reported to have side effects when taking Abraxane. Among them, 25 people (1.32%) have Renal Failure. Time on Abraxane when people have Renal failure :

27. Targeted Nanoparticle A dual Nanoparticle, the targeting ligand allow it to diagnose if a cell is healthy or sick, and bind specifically to the tumorous cell Once inside the cell, the polymeric nanoparticle degrades and the anticancer agent is set free Imaging agent An imaging agent can be added as well Annu. Rev. Biomed. Eng. 2007. Vol. 9, pp. 257–88

28. Challenges • Real-time monitoring of drug distribution, action mechanism and patient’s response • Fast detection of biomarkers at lower limits • Understanding the mechanism of cancer • Diagnosis leading to personalized treatments • Detection of deep tumors • Selective targeting in extremely heterogeneous tissues.