Nanoscale Drug Delivery Systems PDF

Nanoscale Drug Delivery Systems Lifeng Kang 1 Outline Introduction NP examples Gold NPs Quantum dots (QD)s Liposomes and lipid NPs Fabrication Synthesis Surface modification Applications Nanomedicine ba...

Nanoscale Drug Delivery Systems Lifeng Kang 1 Outline Introduction NP examples Gold NPs Quantum dots (QD)s Liposomes and lipid NPs Fabrication Synthesis Surface modification Applications Nanomedicine basics Gene delivery Protein delivery 2 Introduction 3 “There’s plenty of room at the bottom” by Richard Feynman in 1959 "There's Plenty of Room at the Bottom: An Invitation to Enter a New Field of Physics" was a lecture given by physicist Richard Feynman at the annual American Physical Society meeting at Caltech on December 29, 1959. Feynman considered the possibility of direct manipulation of individual atoms as a more robust form of synthetic chemistry than those used at the time. Source: https://upload.wikimedia.org/wikipedia/en/4/42/Richard_Feynman_Nobel.jpg https://www.nature.com/articles/nnano.2009.356 4 The scale of things Things Natural Things Manmade 5 10.18260/1-2--4555 Historical timeline of clinical-stage nanoparticle technologies Genexol-PM (PLAPEG micelle, paclitaxel) Liposome Doxil (stealth liposome, marketed in Korea in described in doxorubicin) approved by 2007 1960s FDA in 1995 Targeted polymeric First published Abraxane (Nabpaclitaxel) NP (BIND-014) dendrimer in approved by FDA in 2005 entered clinical trials 1978 in 2011 Ferumoxide (iron oxide Targeted cyclodextrin- NP) approved as MR containing polymer NP First targeted imaging agent in 1996 (CALAA-01) entered liposome in 1980 clinical trials in 2008 First controlled release polymer Long circulating system for PLGA-PEG NP in 1994 macromolecules in 1976 Nano Lett. 2010, 10(9): 3223–3230. 6 Lipid nanoparticle–mRNA formulations as COVID-19 vaccines. Nature Reviews Materials volume 6, pages1078–1094 (2021) 7 Recombinant hepatitis B vaccine Recombinant hepatitis B vaccine was licensed in the United States in 1986, and was the first licensed vaccine in the United States produced by recombinant DNA technology. Recombinant hepatitis B vaccine is produced by inserting a plasmid containing the gene for hepatitis B surface antigen (HBsAg) into a recombinant strain of common baker’s yeast (Saccharomyces cerevisiae). The antigen is harvested and purified from fermentation cultures of the recombinant yeast containing the gene. https://www.sciencedirect.com/topics/pharmacology-toxicology-and-pharmaceutical-science/recombinant-hepatitis-b-vaccine 8 What is special about nanoscale? Atoms and molecules are generally less than one nanometer, and we study them in chemistry. Condensed matter physics deals with solids with infinite array of bound atoms. Nanoscience deals with the in-between meso-world. Surface to volume ratio. Size-dependent properties. 9 Percentage of surface atoms Full-shell Clusters Total Number Surface of Atoms Atoms (%) Spherical iron nanocrystals 1 Shell 13 92 2 Shell 55 76 3 Shell 147 63 4 Shell 309 52 5 Shell 561 45 7 Shell 1415 35 Surface to volume ratio A 3 nm iron particle has 50% atoms on surface; A 10 nm particle has 20% on surface; A 30 nm particle has only 5% on surface. 10 Nanoscale = high ratio of surface area to volume Click the play button to view the animation. 11 NP examples Gold NPs Quantum dots (QD)s Liposomes and Lipid NPs 12 Metal nanoparticles Pioneer Michael Faraday 1857 his gold colloids 35 Å Gold Nanoparticles and their Convex Hulls AFM of Gold Nanoparticles 13 Source: Wikimedia Commons Melting Point Start from an energy balance; assume the change in internal energy (∆U) and change in entropy per unit mass during melting are independent of temperature ∆ = Deviation of melting point from the bulk value  = 2To / Lr To = Bulk melting point  = Surface tension coefficient for a liquid-solid interface  = Particle density r = Particle radius L = Latent heat of fusion Melting Point depends on particle size!  gold particles For example: The melting point of gold particles decreases dramatically as the particle size gets below 5 nm. Physical Fundamentals of Nanomaterials. DOI: https://doi.org/10.1016/B978-0-12-410417-4.00007-1 14 Surface plasmon resonance (SPR) and localized surface plasmon resonance (LSPR) Localized surface plasmon resonance (LSPR) is SPS Surface plasmon resonance (SPR) is the collective on nanosized structures. oscillation of conduction band electrons that are in https://youtu.be/eVpxn5Cw6YM resonance with the oscillating electric field of incident light, which will produce energetic plasmonic electrons through non-radiative excitation. J. Phys. Chem. B 2003, 107, 668 15 Localized surface plasmon resonance When the resonance is coherent with that of the external optical field, nanoparticles exhibit strong absorption at this wavelength of the external field. Absorption is transferred into heat. 520 nm 14 nm Au nanoparticles UV-vis spectrum 16 Why are gold nanoparticle dispersion red in color? Au nanoparticles 14 nm Au White nanoparticles Light Red light pass 520 nm through! 17 Gold nanoparticles as a quencher Fluorescein isothiocyanate (FITC) is a small organic fluorochrome with an excitation maximum (Ex-Max) at 494 nm and an emission maximum (Em-Max) at 518 nm. 14 nm Au nanoparticles FITC + Au 18 Localized surface plasmon resonance: size dependence Au nanoparticles Surface plasmon resonance of metal nanoparticles red-shifts with increasing nanoparticle sizes. uneven excitation of free electrons for bigger nanoparticles leads to the retardation effect and correspondingly the red-shift Extinction (a.u.) (move to longer wavelength) of the SPR. “retardation effect” Wavelength (nm) Int. Rev. Phys. Chem. 2000, 19, 409 19 Localized surface plasmon resonance: shape effect Extinction Au Nanorods (a.u.) Wavelength (nm) Au nanorods exhibit both transverse and longitudinal surface plasmon resonances (along their two axes). Their longitudinal peaks are very sensitive to their aspect ratio (L/W ratio) (larger ratio leads to longer wavelength) and can cover a wide spectral range. Acc. Chem. Res. 2008, 12, 1578 20 Localized surface plasmon resonance: aggregation state Surface plasmon band red-shifts upon aggregation of nanoparticles due to interparticle coupling. For 14 nm Au nanoparticles, the dispersion color experiences red-to-blue transition during aggregation, which is widely used in Au nanoparticle-based colorimetric biosensors. 21 10.1021/acssensors.3c00287 Quantum dots (QD)s 22 Colors: visible light spectrum Longer wavelength Shorter wavelength Lower energy Higher energy Wavelength of Wi-Fi? 23 Colors: eye color sensitivity & white light 24 Semiconductor quantum dots A quantum dot (QD) is a semiconductor nanostructure that confines the motion of conduction band electrons, valence band holes in all three spatial directions (wiki). Larger QDs emit light with longer wavelength than that of the smaller ones. Semiconductor e- Conduction Band Energy Gap h+ Valence Band 25 Semiconductor quantum dots Photograph of 10 QDs of different sizes Fluorescence Spectra (irradiated by 365nm UV light) (1-10 nm) size increase Emission wavelength of QDs increases with their sizes 26 Quantum size effect When size of the QD is smaller than the exciton bohr radius (electron & hole), energy band gap of the QD exhibits strong quantum size confinement and becomes size-dependent. Electronic Energy Level conduction Energy Level of QDs electron band The band gap of QDs is size-dependent and decreases energy gap with increasing sizes. valence Smaller band gap corresponds to longer wavelength. hole band Small Big Bulk QDs Semiconductor 27 Quantum size effect Quantum Dots Energy Space level increase increase Electron 28 Quantum size effect Electronic energy level of QDs conduction band Ex Em energy gap valence band Small Big Quantum dot emits light when electrons that were excited into the conduction return to valence band and combine with holes. Light that can excite smaller QDs also can excite larger QDs. QD emission color evolves from blue, green to red when its size increases. 29 QD: optical properties Narrow & symmetrical emission Single wavelength excitation of multi-colored QDs Broad absorption profile Superior photo-stability 30 Liposomes and lipid NPs 31 What is a liposome ? Spherical vesicles with a phospholipid bilayer Hydrophilic Hydrophobic Phosphatidylethanolamine What’s the driving force to form this structure? 32 Liposomes (1965) Liposomes are concentric bi-layered vesicles in which an aqueous volume is entirely enclosed by a membranous lipid bilayer mainly composed of natural or synthetic phospholipids Lipid bilayer Surface proteins to target liposome to specific location Alec Bangham in the body (1921–2010) Surface sugars to prevent destruction by the immune system Solid or water- soluble drugs in the core Oil-soluble drugs in the lipid bilayer 33 Phospholipids and cholesterol High concentration up to 2:1 molar ratios of cholesterol to PC. Hydroxyl group of cholesterol oriented towards the aqueous surface and aliphatic chain aligned parallel to the acyl chains in the center of the bilayer 〜 100nm Hydrophilic head Polar molecule Phosphate group Glycerol : Cholesterol backbone Hydrophobic tail : Phospholipid Fatty acids : Anticancer agent Phospholipid 34 The bilayer phospholipids theory Liposomes are formed when thin phospholipid films are hydrated Phospholipid bilayers spontaneously close in on themselves to form sealed compartments, which are called liposomes. https://avantilipids.com/tech-support/liposome-preparation/ 35 Methods for the preparation of liposomes 36 Small unilamellar Large unilamellar vesicles vesicles Multilamellar vesicles 37 Doxil® (FDA 1995) for AIDS-related kaposi sarcoma, multiple myeloma, ovarian cancer (IV) Doxorubicin hydrochloride encapsulated in Stealth®liposomes (100 nm) composed of N- (carbonyl-methoxypolyethylene glycol 2000)-1,2-distearoyl-sn-glycero3-phosphoethanolamine sodium, fully hydrogenated soy phosphatidylcholine, and cholesterol Doxorubicin Liposome Methoxypolyethylene glycol (MPEG) Doxil® – the first FDA-approved nanodrug: lessons learned. J Control Release. 2012;160(2):117 38 Current liposomal drug preparations Type of Agents Examples Anticancer Drugs Duanorubicin, Doxorubicin*, Epirubicin Methotrexate, Cisplatin*, Cytarabin Anti bacterial Triclosan, Clindamycin hydrochloride, Ampicillin, peperacillin, rifamicin Antiviral AZT DNA material cDNA - CFTR* Enzymes Hexosaminidase A Glucocerebrosidase, Peroxidase Radionuclide In-111*, Tc-99m Fungicides Amphotericin B* Vaccines Malaria merozoite, Malaria sporozoite Hepatitis B antigen, Rabies virus glycoprotein *Currently in Clinical Trials or Approved for Clinical Use 39 Liposomes vs lipid nanoparticles Liposomes are spherical vesicles formed mainly by phospholipids and other physiologic lipids, Lipid nanoparticles are solid particles at room and body temperature, consisting of solid lipids (SLN) or a mixture of a solid lipid and a liquid lipid (NLC). Liposomes and lipid nanoparticles are similar by design, but slightly different in composition and function. LNPs are liposome-like structures especially geared towards encapsulating nucleic acids (RNA and DNA). Traditional liposomes include one or more rings of lipid bilayer surrounding an aqueous pocket, but not all LNPs have a contiguous bilayer that would qualify them as lipid vesicles or liposomes. https://www.exeleadbiopharma.com/news/liposomes-and-lipid-nanoparticles-as-delivery-vehicles-for-personalized-medicine 40 Development of lipid nanoparticles using microfluidics https://youtu.be/hUeFnx9iNNQ DOI: 10.1021/acsabm.1c00732 41 https://www.caymanchem.com/news/intro-to-lipid-nanoparticle-formulation#preparation Fabrications 42  Synthesis Nanoparticles Surface Engineering Bio-functionalization 43 44 Bottom up: nanoprecipitation https://www.youtube.com/watch?v=zRLrz-bhAFQ 45 Bottom up: spray-drying 46 Top-down: PRINT® (Particle Replication in Non-wetting Templates) DeSimone, J. UNC 47 DeSimone, J. UNC 48 Bottom Up: self-assembling delivery systems The next advance was to construct materials that would self assemble with drugs to create controlled drug delivery vehicles Self assembly is typically approach using a molecule that has a hydrophilic head and hydrophobic tail to form a shell J. Colloid Interface Sci. 448 (2015) 367–373 49 Self-assembly Amphiphilic molecules: Surfactant Lipid Block-copolymer 50 Liposome formulation Doxorubicin 51 https://www.sigmaaldrich.com/AU/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/transfection-and-gene-editing/liposome-preparation Bottom up: ruby red colloidal gold https://youtu.be/VuLJlT5UDaI 52 Graphene - electrochemical exfoliation https://youtu.be/s51l6KySFU8 53 Surface engineering Water-solubility Colloidal stability Surface-functionality Nanoparticle-cell interaction Bio-distribution and pharmacokinetics 54 Stabilizing mechanism for nanoparticles in aqueous medium Key features of successful surface coating: highly stable at high-salt medium, resistant to non-specific protein bindings, and easy bioconjugation schemes. 55 Charge-stabilized nanoparticles Electrical double layer refers to two parallel layers of charge surrounding the object. The first layer, the surface charge (either positive or negative), comprises ions adsorbed directly onto the object due to a host of chemical interactions. The second layer is composed of ions attracted to the surface charge via the coulomb force, electrically screening the first layer. The Stern Layer is the first (internal) layer of the electric double layer, which forms at a charged surface in an ionic solution. 56 Charge-stabilized nanoparticles Repulsion of surface charges on nanoparticle surface prevents the nanoparticles to aggregate. Increase of salt concentration leads to thinner electrical double layer and thus reduced electrostatic repulsion (poorer colloidal stability). 57 Polymer ligands: steric stabilization Polymer coated As the distance of separation between the particles decreases in nanoparticles the coagulation step, the polymer coatings begin to overlap, which is entropically unfavorable, and thus push the nanoparticles away from each other. Ultimately, it is the crowding of the polymer chains within this overlap volume that produces the steric stabilizing effect. Polymer ligands should have good solubility in aqueous medium. Higher graft density of polymer ligands leads to better stabilization (stronger steric stabilizing effect). 58 Protein corona on nanoparticles: non-specific binding Nanoparticles, when exposed to biological fluid, become coated with proteins and other bio-molecules to form a ‘protein corona’. The non-specific binding form a monolayer on nanoparticle surfaces and has micromolar affinity, and this non- specific binding can be detrimental to the built-in molecular recognition in the nanoparticles. Nat. Nanotechnol. 2009, 4, 577 59 PEGylation: one way to reduce non-specific binding PEGylation: the process by which poly(ethylene glycol) chains are attached to carriers. Poly(ethylene glycol) (PEG) The ability of PEG to reduce non-specific protein binding is believed to result from the preferred (polar) gauche conformation of PEG in water, which offers two hydrogen bond acceptors in ideal distance for hydrogen bonding with water. This conformation leads to extensive hydration in aqueous environments which, along with good conformational flexibility and high chain mobility, causes a steric exclusion effect that prevents the adsorption of proteins. Nat. Rev. Drug Disc. 2003, 2, 214 60 Nanomedicine Basics 61 Nanomedicine DOX was grafted onto the chitosan skeleton via an acid-cleavable hydrazone (hz) bond. DOI 10.1088/0957-4484/25/25/255101 62 Nanocarriers as an emerging platform for cancer therapy The peptide VATANST (STP) can specifically bind with vimentin, which is highly expressed on the osteosarcoma cell surface. The nanoparticle is core–shell structured to protect the loaded DOX during blood flow. The disulfide bonds within the nanoparticles are sensitive to the osteosarcoma microenvironment, which has high glutathione (GSH) levels. Under the enhanced permeability and retention and active tumor targeting effects, the STP-decorated DOX-loaded NPs accumulated in tumor tissues. High GSH levels can rupture disulfide bonds, resulting in the controlled release of DOX, which will cause necrosis of tumor cells. The nanoparticles could increase the tumor inhibition efficiency against osteosarcoma and reduce the side effects of DOX to major organs. 63 https://doi.org/10.1080/10717544.2020.1856221 ADME - Summary ibuprofen