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Details of RareLED ID Assay_1018
Primary information
GENECTSV
NUMBER OF ASSAY61
ASSAY IDs1063746 1063745 1063742 1063741 568361 568360 48397 1063747 1063744 1063743 1343163 261375 1344290 1258853 1343457 448878 254422 254421 254423 1063748 51194 391116 271492 263492 242468 668142 668140 668138 668141 668139 668137 668134 314224 242446 569129 539534 430242 428164 1161922 1344175 391117 668135 391138 391136 391134 391126 198832 162519 161894 156737 83251 14650 1257557 48021 466962 290159 219227 508158 444709 81585 538816
Assay Name1. Inhibition of human Cathepsin V using Z-Phe-Arg-aminomethylcoumarin as substrate,2. Inhibition of human Cathepsin V using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,3. Inhibition of human Cathepsin H using Arg-aminomethylcoumarin as substrate,4. Inhibition of human Cathepsin H using Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,5. Inhibition of human recombinant cathepsin V expressed in Pichia pastoris by Lineweaver-Burk double-reciprocal plots,6. Inhibition of human recombinant cathepsin V expressed in Pichia pastoris,7. Concentration required to inhibit 50% of cysteine protease cathepsin K of human,8. Inhibition of recombinant human Cathepsin L using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,9. Inhibition of human Cathepsin B using Z-Phe-Arg-aminomethylcoumarin as substrate,10. Inhibition of human Cathepsin B using Z-Phe-Arg-aminomethylcoumarin as substrate preincubated for 30 mins followed by substrate addition,11. Inhibition Assay: Recombinant human cathepsin A (residues 29-480, with a C-terminal 10-His tag; R&D Systems, #1049-SE) was proteolytically activated with recombinant human cathepsin L (R&D Systems, #952-CY). Briefly, cathepsin A was incubated at 10 ug/ml with cathepsin L at 1 ug/ml in activation buffer (25 mM 2-(morpholin-4-yl)-ethanesulfonic acid (MES), pH 6.0, containing 5 mM dithiothreitol (DTT)) for 15 min at 37 degrees C. Cathepsin L activity was then stopped by the addition of the cysteine protease inhibitor E-64 (N-(trans-epoxysuccinyl)-L-leucine-4-guanidinobutylamide; Sigma-Aldrich, # E3132; dissolved in activation buffer/DMSO) to a final concentration of 10 uM.The activated cathepsin A was diluted in assay buffer (25 mM MES, pH 5.5, containing 5 mM DTT) and mixed with the test compound (dissolved in assay buffer containing (v/v) 3% DMSO) or, in the control experiments, with the vehicle in a multiple assay plate. After incubation for 15 min at room temperature.,12. Inhibition of human cathepsin V,13. In-Vitro Fluorescence Assay: In order to identify modulators of cathepsin D activity, a continuous enzymatic test was carried out with a synthetic peptide which carries a fluorescent group (MCA=(7-methoxycoumarin-4-yl)acetyl) which is quenched by energy transfer from a Dpn (2,4 dinitrophenyl) group on the same molecule, in Greiner 384-well nb microtitre plates. Cleavage of the peptidic substrate by cathepsin D causes an increase in the fluorescence intensity. In order to determine the efficacy of substances, the time-dependent increase in the fluorescence intensity in the presence of the substance was compared with the time-dependent increase in fluorescence in the absence of substances. The reference substance used was pepstatin A (Sigma-Aldrich). The substrate used was MCAGKPILFFRLK(Dnp)d-R-NH2 (Enzo Life Sciences, Lorrach). The enzyme employed was cathepsin D isolated from the human liver (Sigma-Aldrich) in a final concentration of 1.4 nM. The test was carried out in 100 mM sodium a,14. Inhibition Assay: The activated cathepsin A was diluted in assay buffer (25 mM MES, pH 5.5, containing 5 mM DTT) and mixed with the test compound (dissolved in assay buffer containing (v/v) 3% DMSO) or, in the control experiments, with the vehicle in a multiple assay plate. After incubation for 15 min at room temperature, as substrate then bradykinin carrying an N-terminal (R)Bodipy FL (4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl) label (JPT Peptide Technologies GmbH; dissolved in assay buffer) was added to the mixture. The final concentration of cathepsin A was 833 ng/ml and the final concentration of labeled bradykinin 2 inverted question markM. After incubation for 15 min at room temperature the reaction was stopped by the addition of stop buffer (130 mM 2-(4-(2-hydroxy-ethyl)-piperazin-1-yl)-ethanesulfonic acid, pH 7.4, containing (v/v) 0.013% (R)Triton X-100, 0.13% Coating Reagent 3 (Caliper Life Sciences), 6.5% DMSO and 20 inverted question markM ebelact,15. Inhibition Assay: The activated cathepsin A was diluted in assay buffer (25 mM MES, pH 5.5, containing 5 mM DTT) and mixed with the test compound (dissolved in assay buffer containing (v/v) 3% DMSO) or, in the control experiments, with the vehicle in a multiple assay plate. After incubation for 15 min at room temperature, as substrate then bradykinin carrying an N-terminal(R) Bodipy FL (4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl) label (JPT Peptide Technologies GmbH; dissolved in assay buffer) was added to the mixture. The final concentration of cathepsin A was 833 ng/ml and the final concentration of labeled bradykinin 2 inverted question markM. After incubation for 15 min at room temperature the reaction was stopped by the addition of stop buffer (130 mM 2-(4-(2-hydroxy-ethyl)-piperazin-1-yl)-ethanesulfonic acid, pH 7.4, containing (v/v) 0.013% degrees Triton X-100, 0.13% Coating Reagent 3 (Caliper Life Sciences), 6.5% DMSO and 20 inverted question markM e,16. Inhibition of cathepsin V after 10 to 15 mins by fluorescence assay,17. Inhibitory concentration against recombinant human Cathepsin L at 37 degree C at pH 5.5,18. Inhibitory concentration against recombinant human Cathepsin K at 37 degree C at pH 5.5,19. Inhibitory concentration against recombinant human Cathepsin S at 37 degree C at pH 5.5,20. Inhibition of recombinant human Cathepsin L using Z-Phe-Arg-aminomethylcoumarin as substrate,21. Inhibition of cysteine protease cathepsin K of rat,22. Inhibition of humanized rabbit cathepsin K Tyr61Asp and Val157Leu mutant,23. Inhibition of cathepsin V,24. Inhibition of Cathepsin V,25. Inhibitory concentration against human cathepsin V by fluorescence assay using 2 uM Cbz-Phe-Arg-AMC,26. Competitive inhibition of human recombinant cathepsin V expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,27. Uncompetitive inhibition of human recombinant cathepsin V expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,28. Noncompetitive inhibition of human recombinant cathepsin V expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,29. Competitive inhibition of human recombinant cathepsin L expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,30. Uncompetitive inhibition of human recombinant cathepsin L expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,31. Noncompetitive inhibition of human recombinant cathepsin L expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by Lineweaver-Burk plot analysis,32. Inhibition of human recombinant cathepsin L expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by fluorometric analysis,33. Inhibition of cathepsin V,34. Inhibition of 2 lM Cbz-Phe-Arg-AMC binding to human cathepsin V activity in fluorescence assay with 100 mM NaOAc,35. Inhibition of human cathepsin V,36. Inhibition of human cathepsin V by fluorescence assay,37. Inhibition of cathepsin L2 assessed as inhibition of fluorogenic substrate cleavage,38. Inhibition of cathepsin L2,39. Inhibition of human cathepsin D assessed as cleavage of wtKAHRP fluorogenic substrate,40. Inhibition Assay: 5 inverted question markL of a human cathepsin S enzyme (R&D1183-CY-010) was added to a 96-well plate so as to be 20 ng/well. Next, 10-fold dilution of the test compound (a 10 mM DMSO solution) was serially performed with a buffer solution for assay (a solution of 50 mM sodium methoxide, 250 mM sodium chloride and 5 mM dithiothreitol (DTT), adjusted to pH=4.5 with 1 N hydrochloric acid) six times such that the final concentration becomes 0.1 nM to 10 inverted question markM then, 20 inverted question markL of each of the solutions was added to the wells (the final DMSO concentration was 0.1%), and 25 inverted question markL of synthetic substrate VVR-AMC (3211-V manufactured by PEPTIDE INSTITUTE, INC.) was added thereto such that the final concentration becomes 40 inverted question markM, thereby starting a enzyme reaction. A fluorescence intensity (an excitation wavelength: 380 nm and a fluorescent wavelength: 460 nm) was measured at 37 degrees C. for 10 minute,41. Inhibition of rabbit cathepsin K,42. Inhibition of human recombinant cathepsin V expressed in Pichia pastoris using Z-Phe-Arg-MCA as substrate by fluorometric analysis,43. Drug level in ovariectomized monkey model of osteoporosis at 1 mg/kg, po qd for 1 week measured after 24 hrs of last dose,44. Cmax in ovariectomized monkey model of osteoporosis at 1 mg/kg, po qd for 1 week,45. Inhibition of bone resorption in rhesus monkey bone,46. Inhibition of bone resorption in rabbit bone,47. The inhibitory concentration value was measured on Human renin at 100000 nM,48. In vitro inhibition of HIV-1 protease.,49. Inhibition of viral replication by XTT endpoint,50. The inhibitory concentration value was measured on porcine pepsin at 31000 nM,51. The concentration was measured on HIV-2CBL-20 virus,52. The maximum plasma concentration was measured on rats after oral administration,53. In Vitro Inhibition Assay: In vitro inhibition assay using cathepsin.,54. The inhibitory concentration value was measured on Human cathepsin D at 31000 nM,55. Inhibition of cathepsin G,56. Inhibition of human recombinant cathepsin V,57. In vitro inhibitory activity against bovine Cathepsin D,58. Inhibition of human Cathepsin V,59. Inhibition of human Cathepsin V,60. The concentration was measured on HIV-1 IIIB virus,61. Inhibition of human Cat V
ACTIVITYSubstance BioActivity: 1 Active, 1 Activity <= 1 µM, 1 Tested Substance BioActivity: 1 Active, 1 Activity <= 1 µM, 1 Tested Substance BioActivity: 1 Tested Substance BioActivity: 1 Tested Substance BioActivity: 9 Active, 3 Activity <= 1 µM, 9 Tested Substance BioActivity: 11 Active, 11 Tested Substance BioActivity: 40 Active, 9 Activity <= 1 nM, 39 Activity <= 1 µM, 40 Tested Substance BioActivity: 1 Active, 1 Activity <= 1 µM, 1 Tested Substance BioActivity: 1 Active, 1 Tested Substance BioActivity: 1 Active, 1 Tested Substance BioActivity: 208 Active, 180 Activity <= 1 µM, 208 Tested Substance BioActivity: 2 Active, 1 Activity <= 1 nM, 2 Activity <= 1 µM, 13 Tested Substance BioActivity: 42 Active, 1 Activity <= 1 nM, 38 Activity <= 1 µM, 42 Tested Substance BioActivity: 403 Active, 152 Activity <= 1 µM, 500 Tested Substance BioActivity: 139 Active, 37 Activity <= 1 µM, 139 Tested Substance BioActivity: 1 Active, 1 Tested Substance BioActivity: 32 Active, 28 Activity <= 1 Â