Lead Optimization

Optimizing Leads from Lead Discovery/Hit ID

Kinase Drug Discovery and Development Process


  1. Benefits

  • Flexible: customer-designed assay panels for iterative target and off-target potency (Kd) measurement
  • Measure potency and selectivity in parallel – optimize away from promiscuous inhibitors in real time
  • Discover new target opportunities; extract maximum value from chemical assets
  • Detection of multiple inhibitor types, including type I, type II, and allosteric
  • Reports true thermodynamic affinity values (Kds), not IC50s – directly compare data across kinases
  • Large dynamic range for accurate Kd measurement (<100pM to >10uM)
  • One week turnaround accelerates L.O. and affords rapid SAR feedback
  • Outsourcing frees up internal resources - focus on other high value discovery activities
 

Optimizing Leads from Lead Discovery/Hit ID

Lead optimization is an iterative process that involves modifying hits identified in screening campaigns to optimize several drug-like properties, including potency, selectivity, and pharmacokinetics. KINOMEscan provides a comprehensive portfolio of solutions to support ongoing lead optimization programs. Collect potency and selectivity data in parallel on a schedule synchronized with ongoing medicinal chemistry cycles.

Select from the largest available kinase assay panel to design custom sub-panels for the regular measurement of target/off-target potency data during lead optimization. In parallel, monitor and optimize kinome selectivity by screening lead compounds against our scanMAX/scanEDGE panels or design a customized scanELECT panel of 50-200 assays, which can be screened with 1 week data turnaround as part of an ongoing screening relationship

Add the convenience of outsourced profiling with consistent high quality data and eliminate the expense of failed experiments and costly capital equipment while freeing up internal scientists to focus on other high value discovery activities. In addition to providing personalized screening and selectivity services to support lead optimization, KINOMEscan also offers an expanding suite of investigative study tools that address important structural and kinetic properties of inhibitors, including binding mode (type I/type II) and association/dissociation kinetics/reversibility and enable investigators to further characterize and discern lead compounds and to more clearly interpret data from downstream cellular and in vivo pharmacology models.
 

  1. Services & Solutions

Measure compound binding affinity for any kinase offered by KINOMEscan. Inhibitor binding constants (Kd values) are calculated from duplicate 11-point dose-response curves (plus DMSO control). Measurements are made under optimized conditions that generate true thermodynamic Kd values, as opposed to IC50 values, which complicate inter-kinase potency comparisons.


The largest commercial kinase assay panel available, scanMAX contains a definitive set of 451 kinases covering AGC, CAMK, CMGC, CK1, STE, TK, TKL, lipid, and atypical kinase families, plus important mutant forms and activation state-specific scanMODE assays that provide inhibitor binding mode information early in the drug discovery process.


An economical approach to surveying the human kinome, scanEDGE includes 96 kinases distributed throughout the AGC, CAMK, CMGC, CK1, STE, TK, TKL, lipid, and atypical kinase families, plus important mutant forms and activation state-specific scanMODE assays that provide inhibitor binding mode information early in the drug discovery process.


A flexible 'a la carte' approach to personalized kinase profiling. Select from KINOMEscan's collection of 451 kinase assays and rapidly deploy a customized assay panel of any size comprised only of kinases relevant to your programs.


Employs activated/non-activated & autoinhibited/non-autoinhibited assay pairs to elucidate compound binding mode. An ideal tool for exploring and understanding how kinase activation state affects inhibitor affinity.


Investigative Tools & Mechanism of Action (MOA) Services

Learn more about your kinase inhibitor’s biochemical mechanism of action. KINOMEscan offers a suite of investigative tools that provides a detailed biochemical characterization of the interaction between inhibitors and their targets. The thermodynamic, kinetic, and structural information provided by these tools enables a detailed comparison of inhibitors from common or distinct lead series and facilitates the interpretation of data from downstream cellular and in vivo pharmacology models.

PathHunter® Cell-Based Kinase Screening & Profiling Services - New!

The PathHunter® Cell-Based Kinase Screening and Profiling Service leverages DiscoveRx's proprietary Enzyme Fragment Complementation (EFC) Assay Technology in a whole cell platform enabling customers to interrogate and characterize compounds in a more physiologically relevant context.

Solutions that deliver accurate, precise & reproducible data

KINOMEscan screening partners have long recognized that accurate, precise and reproducible kinase profiling data are critical components of the discovery and development process and are the cornerstone of a successful screening partnership. All KINOMEscan assays undergo thorough validation prior to being released into production and assay performance is continually monitored to ensure data consistency & longitudinal concordance.

  • Data accuracy and precision have been demonstrated in several peer-reviewed publications from both the KINOMEscan team and from our partners.

KdELECT

Quantitative Interaction affinities measured for any kinase/inhibitor combination. Inhibitor binding constants (Kd values) are calculated from duplicate 11-point dose-response curves (plus DMSO control). Measurements are made under optimized conditions that generate true thermodynamic Kd values, as opposed to IC50 values, which can depend on the individual assay conditions (e.g. the ATP concentration). For this reason, KdELECT data facilitate a direct comparison of inhibitor affinity across kinases. 

 
Sample Kd determination data
Binding constant (Kd) determination for the interaction between Compound F and Aurora Kinase B (AURKB). Kd values were calculated by fitting dose-response curves to the Hill binding equation using the Levenberg–Marquardt algorithm. Duplicate Kd values calculated for this interaction were 36.5 nM and 24.3 nM.

Large Dynamic range for accurate affinity measurements

KINOMEscan binding assays use low kinase concentrations (less than 0.1 nM). This feature, coupled with quantitative, precise and ultra-sensitive qPCR readout, affords a large dynamic range (greater than five logs) for the measurement of accurate affinity values (Kd less than 100 pM to greater than 10 uM). These features provide a flexible, sensitive and highly quantitative platform for the identification and discrimination of a wide spectrum of inhibitors.
 

Assay Sensitivity
Assay sensitivity using LOK against AST-487 CHIR-265 CI-1033 and Staurosporine
Binding constant (Kd) determinations for the indicated compounds against LOK demonstrate the broad range (> 5 logs) of interaction affinities quantitatively measured using the KINOMEscan assay platform. Assays are performed at low kinase concentrations (<0.1 nM), which enables the measurement of accurate Kd values in the pM range [click graph to enlarge].

Detect multiple types of kinase inhibitors

Although the majority of kinase inhibitors developed so far have been ATP-competitive type inhibitors that fully or partially overlap the the ATP binding site, there are also examples of non-ATP competitive “allosteric” inhibitors that target the kinase active site and inhibit catalysis by repositioning key catalytic residues. Allosteric inhibition may represent a promising strategy for the development of highly potent & selective compounds. KINOMEscan affords investigators a flexible screening solution for detecting these multiple inhibitor types and to thereby extract the maximum value from their chemical assets. To learn more about the types of allosteric inhibitors detected in KINOMEscan assays, visit our Allosteric Inhibitor page.

Detection of Type I, II & Allosteric Inhibitors
detection of type I, type II and allosteric kinase inhibitors
Binding constant (Kd) determinations were measured for interactions between gefitinib, a known Type I inhibitor and EGFR (Kd = 3.4nM); imatinib, a known Type II inhibitor and KIT (Kd = 7.5nM); and CI-104, a known non-ATP-competitive inhibitor and MEK1 (Kd = 120nM). These data illustrate the diversity of inhibitor types detected in KINOMEscan assays [click graph to enlarge].

Assay quality: Primary screen reproducibility

  • Outstanding screen to screen reproducibility
  • Enables data to be interpreted with confidence

Data Consistency
Profiling of the indicated compounds at 10uM in fourteen independent experiments against 442 kinases over a one year period. Correlation analysis was performed in a pair-wise comparison to calculate the correlation coefficient. The correlation coefficients range from 0.91 to 0.97 with an average of 0.95 [click graph to enlarge].

Assay quality: Z’ values

  • Z’ values measure assay noise and predictive value
  • High Z’ values reproducibly measured across entire assay panel
  • High Z’ values ensure low false positive/negative rates


Assay Quality
Average Z' values and standard deviations were calculated for each kinase based on fourteen control wells per experiment in over 135 independent experiments spanning a period of sixteen months. Average Z' = 0.71 [click graph to enlarge].