title: IKK2 inhibition using TPCA-1-loaded PLGA microparticles attenuates laser-induced choroidal neovascularization and macrophage recruitment.
abstract: The inhibition of NF-κB by genetic deletion or pharmacological inhibition of IKK2 significantly reduces laser-induced choroid neovascularization (CNV). To achieve a sustained and controlled intraocular release of a selective and potent IKK2 inhibitor, 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1) (MW: 279.29), we developed a biodegradable poly-lactide-co-glycolide (PLGA) polymer-delivery system to further investigate the anti-neovascularization effects of IKK2 inhibition and in vivo biosafety using laser-induced CNV mouse model. The solvent-evaporation method produced spherical TPCA-1-loaded PLGA microparticles characterized with a mean diameter of 2.4 ¼m and loading efficiency of 80%. Retrobulbar administration of the TPCA-1-loaded PLGA microparticles maintained a sustained drug level in the retina during the study period. No detectable TPCA-1 level was observed in the untreated contralateral eye. The anti-CNV effect of retrobulbarly administrated TPCA-1-loaded PLGA microparticles was assessed by retinal fluorescein leakage and isolectin staining methods, showing significantly reduced CNV development on day 7 after laser injury. Macrophage infiltration into the laser lesion was attenuated as assayed by choroid/RPE flat-mount staining with anti-F4/80 antibody. Consistently, laser induced expressions of Vegfa and Ccl2 were inhibited by the TPCA-1-loaded PLGA treatment. This TPCA-1 delivery system did not cause any noticeable cellular or functional toxicity to the treated eyes as evaluated by histology and optokinetic reflex (OKR) tests; and no systemic toxicity was observed. We conclude that retrobulbar injection of the small-molecule IKK2 inhibitor TPCA-1, delivered by biodegradable PLGA microparticles, can achieve a sustained and controllable drug release into choroid/retina and attenuate laser-induced CNV development without causing apparent systemic toxicity. Our results suggest a potential clinical application of TPCA-1 delivered by microparticles in treatment of CNV in the patients with age-related macular degeneration and other retinal neovascularization diseases.
authors: Departments of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States of America; James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: Subhash
authors: S
authors: Gaddipati
authors: Departments of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States of America; James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: Qingxian
authors: Q
authors: Lu
authors: Departments of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: Ramesh Babu
authors: RB
authors: Kasetti
authors: James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, Kentucky, United States of America; Department of Chemistry and Biochemistry, University of North Georgia, Oakwood, Georgia, United States of America.
authors: M Clarke
authors: MC
authors: Miller
authors: Beijing Institute of Ophthalmology, Beijing Tong-Ren Eye Center, Capital Medical University, Beijing, China.
authors: Qingjun
authors: Q
authors: Lu
authors: James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, Kentucky, United States of America; Department of Medicine, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: John O
authors: JO
authors: Trent
authors: Departments of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: Henry J
authors: HJ
authors: Kaplan
authors: Departments of Ophthalmology and Visual Sciences, University of Louisville School of Medicine, Louisville, Kentucky, United States of America; James Graham Brown Cancer Center, University of Louisville School of Medicine, Louisville, Kentucky, United States of America.
authors: Qiutang
authors: Q
authors: Li
mesh_terms: D000577:Amides / Q000008:administration & dosage* / Q000737:chemistry; D000818:Animals; D020256:Choroidal Neovascularization / Q000188:drug therapy* / Q000473:pathology; D004195:Disease Models, Animal; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry; D005260:Female; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors*; D019344:Lactic Acid / Q000008:administration & dosage* / Q000737:chemistry; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects* / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011100:Polyglycolic Acid / Q000008:administration & dosage* / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage* / Q000737:chemistry; D013876:Thiophenes / Q000008:administration & dosage* / Q000737:chemistry
chemical_list: D000577:Amides; D004337:Drug Carriers; D047428:Protein Kinase Inhibitors; D013876:Thiophenes; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; C499326:2-((aminocarbonyl)amino)-5-(4-fluorophenyl)-3-thiophenecarboxamide; D051550:I-kappa B Kinase; C496562:Ikbkb protein, mouse