Is the murine dextran sodium sulfate-induced colitis model valid for predicting drug efficacy in IBD? Yvette M Petersen, B. Björkenberg, K. Christjansen, M. Stahlhut, H.D Pedersen and C. Thorkildsen Zealand Pharma A/S Glostrup, Denmark Slide 1 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Introduction Their is no single animal model of intestinal inflammation that mimics the pathogenesis, pathology, and therapeutic response of IBD Dextran sodium sulfate ()-induced colitis model Spontaneous colitis model Primarily used to assess the therapeutic potential of novel therapies in IBD Most widely used experimental model of ulcerative colitis. Slide 2 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
-induced model of colitis Induction Species Mechanism Features Advantages Disadvantages (2-1% w/v), administered in drinking water for days (5-14 days) followed, in some cases, by a period of normal drinking water Primarily mice but also rats Directly damages epithelial cells leading to the activation of the mucosal immune system Body weight loss, colon shortening, bloody diarrhea, histological damage, neutrophil infiltrations, changes in local cytokine concentrations Reproducible, easy administration, cheap Different pathogenesis vs. IBD Inflammation restricted to the colon Slide 3 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Rationale Aminosalicylates and corticosteroids, are used to therapeutically suppress inflammation in IBD. There are, however, no systematic reports documenting beneficial dose dependent effects of aminosalicylates and corticosteroids in the model. CRO s offering the model rarely use aminosalicylates and corticosteroids as reference compounds. Slide 4 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Aim To evaluate the effects of sulfasalazine (aminosalicylate) and dexamethasone (corticosteroid) in CD-1 mice with induced colitis. Slide 5 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Experimental design Study 1: Sacrifice Study 2: Sacrifice 5% in drinking water 1 2 3 4 5 6 7 8 Sulfasalazine (Sigma-Aldrich) (5, 1 and 15 mg/kg/d, p.o., once daily) controls (p.o., once daily) 5% in drinking water 1 2 3 4 5 6 7 8 Dexamethasone (Boehringer Ingelheim ) (.4,.8 and 1.2 mg/kg/d, i.p., once daily) controls (i.p., once daily) Untreated controls were also included in the study Endpoints: Bodyweight, rectal bleeding, stool consistency, colon mass to length ratio, and colon tissue levels of C-reactive protein (CRP), myeloperoxidase (MPO) and proinflammatory cytokines Slide 6 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Bodyweight in CD-1 mice with -induced colitis Change in bodyweight 4 2-2 -4-6 -8 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine Change in bodyweight 4 2-2 -4-6 -8.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone Sulfasalazine had no effect on bodyweight loss Dexamethasone aggravated bodyweight loss Slide 7 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Stool consistency in CD-1 mice with -induced colitis Stool Consistency Normal stools Mild diarrhea 1 3 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine 3.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone Moderate diarrhea Severe diarrhea 2 3 Stool consistency 2 1 Stool consistency 2 1 Median score Sulfasalazine had no effect on diarrhea severity Dexamethasone had no effect on diarrhea severity Slide 8 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colon mass to length in CD-1 mice with -induced colitis 5 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine 5.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone Colon mass/length 4 3 2 1 Colon mass/length 4 3 2 1 Sulfasalazine had no effect on colon mass to length Dexamethasone had no effect on colon mass to length Slide 9 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colonic CRP concentrations in CD-1 mice with -induced colitis CRP (U/g protein) 2 15 1 5 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine CRP (U/ g protein) 35 3 25 2 15 1 5.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone Sulfasalazine had no effect on colonic CRP concentrations Dexamethasone had no effect on colonic CRP concentrations Slide 1 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colonic MPO concentrations in CD-1 mice with -induced colitis 2 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine 175.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone MPO (U/g protein) 15 1 5 MPO (U/ gprotein) 15 125 1 75 5 25 Sulfasalazine had no effect on colonic MPO concentrations Dexamethasone had no effect on colonic MPO concentrations Slide 11 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colonic TNF-α concentrations in CD-1 mice with -induced colitis TNF-α (U/ g protein) 14 12 1 8 6 4 2 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine TNF-α (U/g protein) 14 12 1 8 6 4 2.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone Sulfasalazine increased colonic TNF-α concentrations Dexamethasone increased colonic TNF-α concentrations Slide 12 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colonic IL-1β concentrations in CD-1 mice with -induced colitis 1. 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine 1..4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone IL-1β (ug/ml protein).75.5.25. IL-1β (ug/ml protein).75.5.25. Sulfasalazine had no effect on colonic IL-1β concentrations Dexamethasone decreased colonic IL-1β concentrations Slide 13 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Colonic IL-6 concentrations are increased in CD-1 rats with -induced colitis 2 5 mg/kg/d Sulfasalazine 1 mg/kg/d Sulfasalazine 15 mg/kg/d Sulfasalazine 2.4mg/kg/d Dexamethasone.8mg/kg/d Dexamethasone 1.2 mg/kg/d Dexamethasone IL-6 (U/g protein) 15 1 5 IL-6 (U/ gprotein) 15 1 5 Sulfasalazine had no effect on colonic IL-6 concentrations Dexamethasone increased colonic IL-6 concentrations Slide 14 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Discussion Might better effects been attained : Using another mouse strain or another species? With other dosing regimens? Had other types of ASA or corticosteoroids used? Using another regimen? Slide 15 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Conclusions -induced colitis in CD-1 mice Sulfasalazine treatment had no effect on the majority of colitis endpoints Dexamethasone treatment either had no effect or aggravated the endpoints assessed Slide 16 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation
Missing link: From the Laboratory to Clinical Practice Gastro-intestinal peptides Probiotics Plant-derived substances Proven efficacy in the -induced colitis model Anti-TNF agents Corticosteroids Immunomodulators What is the Predictive Value of the Model? Who is convinced by the results? Aminosalicylates Suboptimal / No efficacy demonstrated in the -induced colitis model Not validated clinically Validated clinically Slide 17 31st May 29 Session Title: Preclinical Treatment of Experimental Intestinal Inflammation