標 準 納 期 2 週 間 を 誇 るKINOMEscan 受 託 試 験 : 報 告 書 から 抽 出 できる 更 なるデータ? 日 本 事 業 担 当 大 嶺 青 河 DiscoveRx Corporation
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1 標 準 納 期 2 週 間 を 誇 るKINOMEscan 受 託 試 験 : 報 告 書 から 抽 出 できる 更 なるデータ? 日 本 事 業 担 当 大 嶺 青 河 DiscoveRx Corporation
2 DiscoveRx Corporation Fremont, CA San Francisco, CA San Diego, CA
3 DiscoveRx Corporation 3 KINOMEscan Ohmine
4 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 4 KINOMEscan Ohmine
5 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 5 KINOMEscan Ohmine
6 KINOMEscan 新 規 ターゲット 情 報 6 KINOMEscan Ohmine
7 KINOMEscan 新 規 ターゲット 情 報 7 KINOMEscan Ohmine
8 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 8 KINOMEscan Ohmine
9 Active-Site Directed Competitive Binding Human kinases tagged with DNA - ~66% expressed as T7 phage fusions (E. coli) - ~33% expressed as NFkB fusions (HEK-293) - Final concentration of target in assays <0.1 nm Ligand immobilized on solid support (e.g. staurosporine) Measure amount of kinase bound to immobilized ligand in the presence and absence of test compound (qpcr readout) Competition No Competition - Test Compound + Test Compound 9 KINOMEscan Ohmine
10 Types of Experiments Primary screens - Single concentration screens in duplicate across full assay panel - Percent of Control (PoC) data reported -100 PoC no hit ; 0 PoC strong hit - Semi-quantitative relationship between PoC and affinity Secondary screens: K d measurements - 11 point dose response curves performed in duplicate - Measure true thermodynamic inhibitor K d s, not IC 50 s Primary Screen Data K d Data 10 KINOMEscan Ohmine
11 Optimal Screening Concentration and Data Interpretation for KINOMEscan Experiments KINOMEscan assays are non-homogeneous Z values are very good but not perfect (median 0.7) Data points have a proportional error Error bars are larger for high PoC values than for low PoC values Implications Screens designed to detect strong hits <35 PoC Do not over-interpret PoC >35 Set screening concentration >3x K d of interest e.g. to identify all K d s < 1 um, screen at 10 um and follow-up on hits (PoC < 35) Differs from typical screening paradigm where screening concentrations set to the K d threshold of interest (values ~50 PoC interpreted) PoC PoC data < 35 most reliable [Inhibitor], nm Carefully define your K d threshold of interest and screen 3-10x higher 11 KINOMEscan Ohmine
12 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 12 KINOMEscan Ohmine
13 Cover Page 13 KINOMEscan Ohmine
14 KINOMEscan Assay Protocol 14 KINOMEscan Ohmine
15 Relationship of K d to PoC Data 15 KINOMEscan Ohmine
16 Box Plot 16 KINOMEscan Ohmine
17 Relationship of K d to PoC Data 3,500 Interactions Queried K d value of 40,000 assigned to non-binders in dose-response experiment K d (nm) Median values show expected theoretical relationship between K d and PoC Low whiskers end at low outlier >35 Min Outlier Max Outlier PoC Bin (10 µm screen) Screens are semi-quantitative plots provide guidance 17 KINOMEscan Ohmine
18 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 18 KINOMEscan Ohmine
19 Data Generated in KINOMEscan Primary Reports 19 KINOMEscan Ohmine
20 Data Generated in KINOMEscan K d Reports 20 KINOMEscan Ohmine
21 Dose-Down in K d Experiments Lowering the top dose concentration allows us to generate a dose response curve with appropriate upper and lower baselines 21 KINOMEscan Ohmine
22 Selectivity Score: Compound Selectivity scanmax and scanedge screens will include a table with selectivity score analysis A compound selectivity score (S-score) is a quantitative measure of compound selectivity S-scores are calculated for each compound using the following equation S(35) = # hits <35% of control # kinases* tested * distinct (non-mutant) kinases 22 KINOMEscan Ohmine
23 Selectivity Score: Overall Selectivity Primary screen S-score data provides a nice view of overall compound selectivity KINOMEscan reports contain a plot of 38 kinase inhibitors to provide a general reference point for compound selectivity 23 KINOMEscan Ohmine
24 Relative Selectivity Intended target Relative selectivity is also an important inhibitor characteristic - What is the relative potency for the intended target over any off-targets These plots generated using K d data Relative selectivity is informative, but also must be interpreted carefully if the intended target has large potency offsets in cell-based assays (e.g. high ATP affinity) 24 KINOMEscan Ohmine
25 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 25 KINOMEscan Ohmine
26 Mutant Kinase Data scanmax screens contain 59 clinically relevant mutants Data provided by these assays can biochemically informative for inhibitor binding 26 KINOMEscan Ohmine
27 scanmode: Found inside scanmax ABL1 pairs RTKs scanmax screens contain all of the assays that comprise our scanmode offering - ABL1 phosphorylated and non-phosphorylated Type I vs Type II inhibitors - Autoinhibited/non-autoinhibited PDGFR family RTKs activation state-dependent inhibition 27 KINOMEscan Ohmine
28 Overview Panel Expansion News Overview of KINOMEscan technology - Refresher on efficient strategies for designing KINOMEscan experiments In-depth review of the KINOMEscan Study Report layout and information contained within - Primary Screen and K d Reports - KINOMEscan and BROMOscan Reports Data analysis overview: - Types of data generated - Under-utilized data provided by scanmax screens - Data visualization tools: TREEspot 28 KINOMEscan Ohmine
29 TREEspot Compound Profile Visualization Tool scanmax, scanedge, scantk screens contain TREEspot data visualization images 29 KINOMEscan Ohmine
30 TREEspot Create Publication Quality Images 30 KINOMEscan Ohmine
31 TREEspot Visualize Compound Potency 100 nm 1000 nm nm 31 KINOMEscan Ohmine
32 Contact Seiga Ohmine, PhD 日 本 事 業 担 当 大 嶺 青 河 内 線 :123 コスモバイオ( 株 ) 創 薬 支 援 サービスグループ discoverx@cosmobio.co.jp
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