Colitis model case study

Typical screening cascade in an integrated program in inflammation

Oncodesign has over 25 years of experience in pharmacology services, initially in oncology but then extended to inflammatory diseases. A large range of in vivo models are available or can be developed for specific request.

Here, our client wanted to develop a new kinase inhibitor used in treatment of Inflammatory Bowel Disease. Oncodesign developed the screening cascade which allowed to deliver preclinical candidates:

  • Cellular assay
  • Full blood in vitro assay
  • Early ADME
  • PK/PD
  • In vivo validation

We show the data of one of the selected candidate ODS’002 , which was prepared and tested via multiple steps.

This case study is a concrete example of our service offering INPACT PoC Inflammation .

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In vitro screening

Binding inhibition alphaLISA assay was performed to validate ODS’002 activity towards the desired target.

Human recombinant kinase target inhibition - binding assay in alphaLISA

Legend: Human recombinant kinase target inhibition – binding assay in alphaLISA – IC50 = 0.2nM

 

Subsequently, cellular assay permitted to evaluate the capacity of ODS’002 to inhibit L18-MDP-induced TNFα expression. The assay was validated on mouse, rat and human PBMCs.

 

L18-MDP-induced TNFa inhibition - mouse macrophage cell assay

Legend: L18-MDP-induced TNFa inhibition – Mouse macrophages cell assay. IC50 = 5nM

L18-MDP-induced TNFa inhibition - ex-vivo target engagement - human whole blood assay

Legend: L18-MDP-induced TNFα inhibition –Ex-vivo target engagement – Human whole blood assay. IC50 = 3nM

Early ADME evaluation

After the initial in vitro ADME was validated, the pharmacokinetic profile was assessed. Here is the profile of ODS’002 in rats allowing to calculate its bioavailability.

 

Rat pharmacokinetics

Legend: Rat pharmacokinetics (IV clearance and PO biodistribution, with a Tmax of around 2h) – Bioavailability = 85% (ratio oral DNAUC / IV DNAUC)
DNAUC: dose-normalized area under the curve

 

To complement the profile, PK/PD relationship in mice was determined. The ODS’002 test compound was delivered PO , followed by L18-MDP induction of TNFα expression (IV at T+1h). The quantification of TNFα expression is performed at T+2h (PO Tmax).

Mouse PK/PD relationship - acute admnistration

Legend: Mouse pharmacokinetics/pharmacodynamics relationship – Acute administration
In-vivo target engagement IC50 ~0.1 mg/kg

In vivo validation, Anti-CTLA4 and DSS-induced chronic colitis model in mouse

Several readouts are used to demonstrate the therapeutic effect of test compounds (mouse weight monitoring; colonic target gene expression; histology for disease scoring; goblet cells & mucus quantification; epithelial thickness of colonic mucosa).

 

Colitis model - combination of anti-CTLA-4 and DSS

Legend: Colitis model /combination of anti-CTLA-4 and DSS (Dextran Sodium Sulfate), a chemical irritant. ODS’002 protects the colon from inflammation-driven epithelial thickening.

 

This case study is a concrete example of our service offering INPACT PoC Inflammation . Get in touch to see how we can impact your research program.

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