Modified Liposomes with Biomaterials Delivery Applications
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1 Modified Liposomes with Biomaterials Delivery Applications Dr. Rahau Shirazi ACS Meeting Newcastle, Australia November 8th, 2013
2 utline Motivation and Background Focus Future Direction
3 Liposomes Delivery Vehicles Efficient Vector: Encapsulate, Deliver, Release Nonviral vs. viral vectors: Ease of production Unlimited Encapsulation Capacity Safety Efficiency Modified Liposomes Tailored to enhance efficiency Encapsulate (Cationic ph Sensitivity) Protect (e.g. xidative Damage, Enzymatic Degradation, Aggregation in Serum) Deliver (Enhance Circulation, Controlled Release/Environmentally responsive, Increase Bioavailability)
4 Dry Lipid Film Hydration Incubation Liposomes Development Nonviral Gene Delivery Vectors Agitation Sonication Extrusion Membrane Contain Mixtures of Lipids (Charged, Neutral, Bioactive,..) Cationic LiposomeNucleic acid selfassembly Electrostatic interactions Positively Charged Liposomes 5 µm nm Cationic Lipid DNA Counterion Release is Driving Force of Formation CL DNA Complexes
5 Lipid Bilayer Poly (Ethylene Glycol)Cationic Liposomes Encapsulation PEGylation: Delivery in vivo Stability of Liposomes Circulation lifetime Reduce membrane interactions Modified Liposomes Control Release / disassembly N N DNA Complexes with CL DNA nn CL nn N CL nt A phsensitive linker, (acylhydrazonebased PEG2000lipid): Stable at neutral ph (blood) triggered at ph 5 to 4, (late endosomes) Chan, C.L.; Majzoub, R. N.; Shirazi, R. S; Ewert, K. K.; Chen, Y.J.; Liang, K. S.; Safinya, C. R.: Biomaterials 2012
6 Stimuli Responsive Cationic Lipids N H N H H TBTU, DIEA N 2 H 4 hydrate ( 20 x excess) NH N 2 H Cl H 3 N Mature endosome acidifies its contents cleaves the PEG chains closer contact between the membranes of complex H n and endosomes PhI(Ac) 2, cat. n N DB Alahydrazide ("Tail") mpeg2000ch ("PEG") ph 5 CH 2 Cl 2 /MeH mol. sieves N H N N H n acidlabile HPEG2Klipid ("PL") Ref. Chan, C.L.; Majzoub, R. N.; Shirazi, R. S; Ewert, K. K.; Chen, Y.J.; Liang, K. S.; Safinya, C. R.: Biomaterials 2012
7 Stimuli Responsive Cationic Lipids Ref.: Chan, C.L.; Majzoub, R.; Shirazi, R. S; Ewert, K.; Chen, Y.; Liang S.; Safinya, C. Biomaterials 2012 Livecell imaging of transfection by PEGylated CL DNA complexes: DIC and fluorescence micrographs of mouse Lcells incubated with doubly labeled PEGylated CL DNA complexes. (A, B) Images for complexes prepared with the acidstable PEG2Klipid. (C, D) Images for complexes prepared with the acidlabile HPEG2Klipid.
8 utline Motivation and Background Focus Future Direction
9 Biomaterials for NanoDelivery: Hydrophobic: Protected in lipid bilayers Hydrophilic: Sandwiched between lipid bilayers Amphiphilic: rient accordingly LiposomeEncapsulated Biomaterials Characterization: Microscopy Structural Characterisation (ptical, ESEM, STEM) Fluorescent Assays, Nanodrop, UVVis Encapsulation Ability Bioassays Efficiency and toxicity Gene Delivery in vivo
10 Development of Modified Liposomes Liposomes : Synthetic and Natural Sources Synthetic: Natural : Cationic DTAP, DCCholesterol ph Sensitive DDAP, CHEMs Neutral DPE, DPC PEGylation PEG2000PE (18:0, 14:0) CAEP (ADM12) lympic Krill oil (SD238) PGC 80 ICF8 18:1 TAP (DTAP) DCCholesterol HCl 18:1 DAP (DDAP) 18:1 (Δ9Cis) PC (DPC) Cholesteryl hemisuccinate CHEMS 18:1 (Δ9Cis) PE (DPE)
11 ESEM Development Matters Agitation (top) vs Extrusion (below) CAEP (ADM12)
12 ESEM Natural State 746 nm lympic Krill oil (SD238) PGC 80 ICF8
13 ESEM Natural State DCChol:DPE:PEG 2000 PE 18:0
14 Liposomes STEM Negative Staining Pegylated Cationic Liposomes DTAPDPE:PEG2000PE14:0
15 Liposomes STEM Negative Staining DCChol:DPE:PEG 2000 PE 18:0
16 Liposomes as NanoProtectors STEM Negative Staining Unprotected Astaxanthin Liposome Encapsulated Astaxanthin
17 Liposomes Target Gene Delivery Unique Cationic Stealth Liposomes Specific CpGDN sequences with Immunostimulatory adjuvant activity mediated through TollLike Receptor 9 Encapsulation/protect CpG ligodeoxynucleotides (CpGDN) Nuclease degradation Enhance Bioactivity Unique Phosphatidylglyceroylalkylamine (PGAA) based lipids isolated from thermophilic bacteria Meiothermus silvanus Synergistic Effects of Strong Encapsulation Ability Combined with Specific Bioactivity of PGAA
18 utline Motivation and Background Focus and Achievements Conclusion
19 Liposomes encapsulate hydrophobic nanomaterial within their hydrophobic bilayer, and hydrophilic guests in the larger interior. 'stealth /PEGylated liposomes developed for drugdelivery applications can carry specific peptides, polymers, bioactive materials. Cationic liposome NA complexes: onionlike structure with NA sandwiched between cationic membranes. Stimuli responsive liposomes aid control release of biomaterials in stimulus media (e.g. HPEG2000)
20 Questions? Cyrus Safinya et al. Nature 489, , 2012
21 Thank you ACS Meeting IBT Group Bradley Williams Mallaghan Institute of Medical Research Lipid Team Gavin Painter Carbohydrate Chemistry Karen Johnston Group
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