An efficient and low immunostimulatory nanoparticle formulation for systemic siRNA delivery to the tumor.
We have developed a nanoparticle formulation [liposomes-protamine-hyaluronic acid nanoparticle (LPH-NP)] for systemically delivering siRNA into the tumor. The LPH-NP was prepared in a self-assembling process. Briefly, protamine and a mixture of siRNA and hyaluronic acid were mixed to prepare a negatively charged complex. Then, cationic liposomes were added to coat the complex with lipids via charge-charge interaction to prepare the LPH-NP. The LPH-NP was further modified by DSPE-PEG or DSPE-PEG-anisamide by the post-insertion method. Anisamide is a targeting ligand for the sigma receptor over-expressed in the B16F10 melanoma cells. The particle size, zeta potential and siRNA encapsulation efficiency of the formulation were approximately 115 nm, +25 mV and 90%, respectively. Luciferase siRNA was used to evaluate the gene silencing activity in the B16F10 cells, which were stably transduced with a luciferase gene. The targeted LPH-NP (PEGylated with ligand) silenced 80% of luciferase activity in the metastatic B16F10 tumor in the lung after a single i.v. injection (0.15 mg siRNA/kg). The targeted LPH-NP also showed very little immunotoxicity in a wide dose range (0.15-1.2 mg siRNA/kg), while the previously published formulation, LPD-NP (liposome-protamine-DNA nanoparticle), had a much narrow therapeutic window (0.15-0.45 mg/kg).
Chono
Sumio
S
Division of Molecular Pharmaceutics, School of Pharmacy, University of North Carolina, 2316 Kerr Hall, 311 Pharmacy Lane, Chapel Hill, NC 27599, USA.
Li
Shyh-Dar
SD
Conwell
Christine C
CC
Huang
Leaf
L
D000818:Animals; D045744:Cell Line, Tumor; D016207:Cytokines / Q000096:biosynthesis; D020868:Gene Silencing; D006820:Hyaluronic Acid / Q000138:chemical synthesis / Q000737:chemistry; D007114:Immunization; D008081:Liposomes / Q000138:chemical synthesis; D008156:Luciferases / Q000235:genetics; D051379:Mice; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry / Q000633:toxicity*; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics; D010316:Particle Size; D011092:Polyethylene Glycols / Q000138:chemical synthesis; D011479:Protamines / Q000138:chemical synthesis / Q000737:chemistry; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000235:genetics; D014162:Transfection
D016207:Cytokines; D008081:Liposomes; D011479:Protamines; D034741:RNA, Small Interfering; D011092:Polyethylene Glycols; D006820:Hyaluronic Acid; D008156:Luciferases