Development of a novel AAK1 inhibitor via Kinobeads-based screening.

A chemical proteomics approach using Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor-immobilized sepharose (TIM-063-Kinobeads) identified main targets such as CaMKKα/1 and ß/2, and potential off-target kinases, including AP2-associated protein kinase 1 (AAK1), as TIM-063 interactants. Because TIM-063 interacted with the AAK1 catalytic domain and inhibited its enzymatic activity moderately (IC50 = 8.51 µM), we attempted to identify potential AAK1 inhibitors from TIM-063-derivatives and found a novel AAK1 inhibitor, TIM-098a (11-amino-2-hydroxy-7H-benzo[de]benzo[4,5]imidazo[2,1-a]isoquinolin-7-one) which is more potent (IC50 = 0.24 µM) than TIM-063 without any inhibitory activity against CaMKK isoforms and a relative AAK1-selectivity among the Numb-associated kinases family. TIM-098a could inhibit AAK1 activity in transfected cultured cells (IC50 = 0.87 µM), indicating cell-membrane permeability of the compound. Overexpression of AAK1 in HeLa cells significantly reduced the number of early endosomes, which was blocked by treatment with 10 µM TIM-098a. These results indicate TIM-063-Kinobeads-based chemical proteomics is efficient for identifying off-target kinases and re-evaluating the kinase inhibitor (TIM-063), leading to the successful development of a novel inhibitory compound (TIM-098a) for AAK1, which could be a molecular probe for AAK1. TIM-098a may be a promising lead compound for a more potent, selective and therapeutically useful AAK1 inhibitor.

Anti-VEGF Therapy and Retinal Photocoagulation to Prevent Recurrence of Central Retinal Vein Occlusion: Two Case Reports of Young Patients.

Case 1: An 18-year-old man. On initial examination, he was diagnosed with central retinal vein occlusion (CRVO) due to optic papillitis. He had no previous systemic and ocular medical history. His best corrected visual acuity was 20/20 for the right eye, but macular edema accompanied by serous retinal detachment was observed about 2 months after the initial examination. Intravitreal anti-VEGF injection was performed, and the symptoms improved. Then, additional photocoagulation was applied to the retinal nonperfusion area, to maintain its normal state. Case 2: A 36-year-old man. He was diagnosed with neovascular glaucoma associated with CRVO in the right eye by his previous physician. Panretinal photocoagulation and intravitreal anti-VEGF injection were performed under maximum-tolerated medical therapy for the right eye. He had diabetes. On initial examination, his visual acuity was 20/100 and his intraocular pressure was 19mmHg. Macular edema and iris neovascularization recurred half a year later, so intravitreal anti-VEGF injection and additional photocoagulation were applied to nonperfusion area. After that, both macular edema and iris neovascularization have settled down. Conclusions: Intravitreal anti-VEGF injection is effective in young patients with CRVO. Also, photocoagulation to nonperfusion area in addition to the continuous treatment with intravitreal anti-VEGF injection seems effective for preventing recurrence.

Ultrasonic Lateral Displacement Sensor for Health Monitoring in Seismically Isolated Buildings.

An ultrasonic lateral displacement sensor utilizing air-coupled ultrasound transducers is proposed. The normally-distributed far field of an ultrasound transducer in a lateral direction is taken advantage of for measuring lateral displacement. The measurement system is composed of several air-coupled ultrasound transducers as a receiver and several transmitters. The transmitters are immobilized at a fixed point, whereas the receiver set-up is separately arranged on the opposite side. In order to improve measurement accuracy, a correction method that utilizes polynomial approximation is introduced. The difference between the corrected lateral displacement and the reference displacement is estimated to be 0.2 mm at maximum for the two transmitters system. A good responsiveness is demonstrated by conducting a dynamic response experiment. When five transmitters are arranged, their measurement range is easily extended up to ±60 mm with an accuracy of 0.7 mm. In both cases, the fluctuations to the measurement ranges show less than 1%. These results indicate that the developed sensor system is useful for measuring relative lateral displacement of a seismically isolated building in the field of structural health monitoring.