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Discovery of Novel ROCK1 Inhibitors via Integrated Virtual Screening Strategy and Bioassays.

Shen M, Tian S, Pan P, Sun H, Li D, Li Y, Zhou H, Li C, Lee SM, Hou T - Sci Rep (2015)

Bottom Line: In this study, a novel integrated virtual screening protocol by combining molecular docking and pharmacophore mapping based on multiple ROCK1 crystal structures was utilized to screen the ChemBridge database for discovering potential inhibitors of ROCK1.Among the 38 tested compounds, seven of them exhibited significant inhibitory activities of ROCK1 (IC50 < 10 μM) and the most potent one (compound TS-f22) with the novel scaffold of 4-Phenyl-1H-pyrrolo [2,3-b] pyridine had an IC50 of 480 nM.Then, the structure-activity relationships of 41 analogues of TS-f22 were examined.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.

ABSTRACT
Rho-associated kinases (ROCKs) have been regarded as promising drug targets for the treatment of cardiovascular diseases, nervous system diseases and cancers. In this study, a novel integrated virtual screening protocol by combining molecular docking and pharmacophore mapping based on multiple ROCK1 crystal structures was utilized to screen the ChemBridge database for discovering potential inhibitors of ROCK1. Among the 38 tested compounds, seven of them exhibited significant inhibitory activities of ROCK1 (IC50 < 10 μM) and the most potent one (compound TS-f22) with the novel scaffold of 4-Phenyl-1H-pyrrolo [2,3-b] pyridine had an IC50 of 480 nM. Then, the structure-activity relationships of 41 analogues of TS-f22 were examined. Two potent inhibitors were proven effective in inhibiting the phosphorylation of the downstream target in the ROCK signaling pathway in vitro and protecting atorvastatin-induced cerebral hemorrhage in vivo. The high hit rate (28.95%) suggested that the integrated virtual screening strategy was quite reliable and could be used as a powerful tool for identifying promising active compounds for targets of interest.

No MeSH data available.


Related in: MedlinePlus

The protective effects of TS-15 and TS-40 against atorvastatin-induced cerebral hemorrhage in zebrafish.(A,a and Aa) the embryos treated with 0.2% DMSO served as the normal control group; (B,b and Bb; C,c and Cc; D,d and Dd; E,e and Ee) the embryos were pretreated with either 0.2% DMSO, Y27632 (10 mM), TS-15 (20 mM) or TS-40 (20 mM) for 2.0 hours and replaced with 1 mM atorvastatin for 24 hours. Homozygous double transgenic zebrafish, the red fluorescence was the Tg (gata1: dsRed) sd2 (A–E), the green fluorescence was Tg (fli1a: EGFP) y1 (a–e), and the third column was the overlapping photos of the first two columns (Aa, Bb, Cc, Dd and Ee). The arrows indicated the erythrocyte accumulation in cerebral hemorrhage region in zebrafish head.
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f9: The protective effects of TS-15 and TS-40 against atorvastatin-induced cerebral hemorrhage in zebrafish.(A,a and Aa) the embryos treated with 0.2% DMSO served as the normal control group; (B,b and Bb; C,c and Cc; D,d and Dd; E,e and Ee) the embryos were pretreated with either 0.2% DMSO, Y27632 (10 mM), TS-15 (20 mM) or TS-40 (20 mM) for 2.0 hours and replaced with 1 mM atorvastatin for 24 hours. Homozygous double transgenic zebrafish, the red fluorescence was the Tg (gata1: dsRed) sd2 (A–E), the green fluorescence was Tg (fli1a: EGFP) y1 (a–e), and the third column was the overlapping photos of the first two columns (Aa, Bb, Cc, Dd and Ee). The arrows indicated the erythrocyte accumulation in cerebral hemorrhage region in zebrafish head.

Mentions: Atorvastatin, a small molecule drug, has been reported to induce cerebral hemorrhage in zebrafish through the disruption of cell-cell junctions4362. The inhibitory effects of TS-15 and TS-40 against cerebral bleeding were examined in this in vivo cerebral hemorrhage system. The healthy control group displayed normal phenotype without hemorrhage occurrence (Fig. 9A) while the atorvastatin-induced disease model group displayed significant cerebral hemorrhage (Fig. 9B). As for the positive control group and the other experimental groups, the hemorrhage changes were obviously abated by the pretreatment of Y27632 (Fig. 9C), TS-15 (Fig. 9D) or TS-40 (Fig. 9E), when compared with the disease model group.


Discovery of Novel ROCK1 Inhibitors via Integrated Virtual Screening Strategy and Bioassays.

Shen M, Tian S, Pan P, Sun H, Li D, Li Y, Zhou H, Li C, Lee SM, Hou T - Sci Rep (2015)

The protective effects of TS-15 and TS-40 against atorvastatin-induced cerebral hemorrhage in zebrafish.(A,a and Aa) the embryos treated with 0.2% DMSO served as the normal control group; (B,b and Bb; C,c and Cc; D,d and Dd; E,e and Ee) the embryos were pretreated with either 0.2% DMSO, Y27632 (10 mM), TS-15 (20 mM) or TS-40 (20 mM) for 2.0 hours and replaced with 1 mM atorvastatin for 24 hours. Homozygous double transgenic zebrafish, the red fluorescence was the Tg (gata1: dsRed) sd2 (A–E), the green fluorescence was Tg (fli1a: EGFP) y1 (a–e), and the third column was the overlapping photos of the first two columns (Aa, Bb, Cc, Dd and Ee). The arrows indicated the erythrocyte accumulation in cerebral hemorrhage region in zebrafish head.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4645114&req=5

f9: The protective effects of TS-15 and TS-40 against atorvastatin-induced cerebral hemorrhage in zebrafish.(A,a and Aa) the embryos treated with 0.2% DMSO served as the normal control group; (B,b and Bb; C,c and Cc; D,d and Dd; E,e and Ee) the embryos were pretreated with either 0.2% DMSO, Y27632 (10 mM), TS-15 (20 mM) or TS-40 (20 mM) for 2.0 hours and replaced with 1 mM atorvastatin for 24 hours. Homozygous double transgenic zebrafish, the red fluorescence was the Tg (gata1: dsRed) sd2 (A–E), the green fluorescence was Tg (fli1a: EGFP) y1 (a–e), and the third column was the overlapping photos of the first two columns (Aa, Bb, Cc, Dd and Ee). The arrows indicated the erythrocyte accumulation in cerebral hemorrhage region in zebrafish head.
Mentions: Atorvastatin, a small molecule drug, has been reported to induce cerebral hemorrhage in zebrafish through the disruption of cell-cell junctions4362. The inhibitory effects of TS-15 and TS-40 against cerebral bleeding were examined in this in vivo cerebral hemorrhage system. The healthy control group displayed normal phenotype without hemorrhage occurrence (Fig. 9A) while the atorvastatin-induced disease model group displayed significant cerebral hemorrhage (Fig. 9B). As for the positive control group and the other experimental groups, the hemorrhage changes were obviously abated by the pretreatment of Y27632 (Fig. 9C), TS-15 (Fig. 9D) or TS-40 (Fig. 9E), when compared with the disease model group.

Bottom Line: In this study, a novel integrated virtual screening protocol by combining molecular docking and pharmacophore mapping based on multiple ROCK1 crystal structures was utilized to screen the ChemBridge database for discovering potential inhibitors of ROCK1.Among the 38 tested compounds, seven of them exhibited significant inhibitory activities of ROCK1 (IC50 < 10 μM) and the most potent one (compound TS-f22) with the novel scaffold of 4-Phenyl-1H-pyrrolo [2,3-b] pyridine had an IC50 of 480 nM.Then, the structure-activity relationships of 41 analogues of TS-f22 were examined.

View Article: PubMed Central - PubMed

Affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang 310058, China.

ABSTRACT
Rho-associated kinases (ROCKs) have been regarded as promising drug targets for the treatment of cardiovascular diseases, nervous system diseases and cancers. In this study, a novel integrated virtual screening protocol by combining molecular docking and pharmacophore mapping based on multiple ROCK1 crystal structures was utilized to screen the ChemBridge database for discovering potential inhibitors of ROCK1. Among the 38 tested compounds, seven of them exhibited significant inhibitory activities of ROCK1 (IC50 < 10 μM) and the most potent one (compound TS-f22) with the novel scaffold of 4-Phenyl-1H-pyrrolo [2,3-b] pyridine had an IC50 of 480 nM. Then, the structure-activity relationships of 41 analogues of TS-f22 were examined. Two potent inhibitors were proven effective in inhibiting the phosphorylation of the downstream target in the ROCK signaling pathway in vitro and protecting atorvastatin-induced cerebral hemorrhage in vivo. The high hit rate (28.95%) suggested that the integrated virtual screening strategy was quite reliable and could be used as a powerful tool for identifying promising active compounds for targets of interest.

No MeSH data available.


Related in: MedlinePlus