TMD’s pre-clinical services evolved from our understanding of the drug mechanisms related to cancer biology. At the pre-clinical level, we offer several sophisticated technologies to support multiple exploratory intentions.
 

Pre-Clinical Compound Development Support
	Proof of disease involvement through target modulation by siRNA, target inhibition, target overexpression, etc. Measurement of target modulation effect by apoptosis, inhibition of cell growth, etc.	Measurement of the effect of drug on target by cDNA Microarrays and Pathway Profiling by IPA	Expression level of target confirmation by qRT-PCR Incidence, specificity & localization of target in disease and normal cell types by Tissue Microarray (TMA) Identification of efficacious cocktails by rationally designed drug combinations Differentiation from competitive compounds by screening against cell line models of innate & acquired resistant	Measurement of target inhibition and dosing In vivo by IHC, ELISA and qWestern Quantitative tumor measurements in vivo of target inhibition and dosing by image analysis

Examples of Pre-clinical Projects Completed at TMD:

• Generated tumor cell line models of acquired resistance to client and market leading ErbB1/ErbB2 inhibitor
  • Cross treatment of therapeutic-specific resistant lines with client inhibitor and competitive inhibitor. 
  • Analyze whether resistance to a particular inhibitor is conserved among similar compounds or
    
  • Probe for alternative mechanism that can be exploited to differentiate similar compounds. 
    
    
• Compared gene expression signatures of drug treated (client and competitive) and resistant cell lines using an Affymetrix microarray. 
  • Analyze and identify biomarkers using signaling and metabolic pathway knowledge database
    
  • Confirm microarray results using quantitative Real-Time PCR. 
    
  • TMD can subsequently create low density, fluidics-based cards to profile small panels of genes (< 24) by quantitative Real-Time PCR. 
    
    
• Growth and apoptosis measurement of tumor cell lines treated with client compound and other drug combinations. 
  • Perform IC50 assays for single drug treatment and combination treatments.
    
    
• Analysis of the effect of an ErbB1/ErbB2 inhibitor on AMPK signaling in human cardiomyocytes.
  • AMPK mobilizes ATP stores in the stressed/failing heart.  Therapeutics that inhibit AMPK may potentially be associated with cardiac toxicity in the clinic.
    
• Analysis of the effect of an ErbB1/ErbB2 inhibitor on AMPK signaling in human tumor cell lines.
  • Activation of the AMPK pathway in tumor cells can inhibit AKT/mTOR signaling and may mediate some of the anti-tumor affects of ErbB1/ErbB2 inhibitors. 
    
• IHC staining and image analysis of a panel of biomarkers in a comparative xenograft study (client ErbB1/ErbB2 inhibitor vs lead competitor). 
  • Using IHC combined with image analysis, a quantitative assessment of target and pathway inhibition at different drug doses and timepoints was established.