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Sulfonamide inhibitors of α2β1 integrin reveal the essential role of collagen receptors in in vivo models of inflammation.

Nissinen L, Ojala M, Langen B, Dost R, Pihlavisto M, Käpylä J, Marjamäki A, Heino J - Pharmacol Res Perspect (2015)

Bottom Line: Since α2β1 integrin is abundantly expressed on various inflammation-associated cells, we tested whether recently developed α2β1 blocking sulfonamides have anti-inflammatory properties.Integrin α2β1 inhibitors were shown to reduce the signs of inflammation in arachidonic acid-induced ear edema, PAF stimulated air pouch, ovalbumin-induced skin hypersensitivity, adjuvant arthritis, and collagen-induced arthritis.Thus, the experiments also revealed fundamental differences in the action of nonactivated and activated α2β1 integrins in inflammation when compared to thrombosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Turku 20014, Turku, Finland ; Biotie Therapies Corp Turku, Finland.

ABSTRACT
Small molecule inhibitors of α2β1 integrin, a major cellular collagen receptor, have been reported to inhibit platelet function, kidney injury, and angiogenesis. Since α2β1 integrin is abundantly expressed on various inflammation-associated cells, we tested whether recently developed α2β1 blocking sulfonamides have anti-inflammatory properties. Integrin α2β1 inhibitors were shown to reduce the signs of inflammation in arachidonic acid-induced ear edema, PAF stimulated air pouch, ovalbumin-induced skin hypersensitivity, adjuvant arthritis, and collagen-induced arthritis. Thus, these sulfonamides are potential drugs for acute and allergic inflammation, hypersensitivity, and arthritis. One sulfonamide with potent anti-inflammatory activity has previously been reported to be selective for activated integrins, but not to inhibit platelet function. Thus, the experiments also revealed fundamental differences in the action of nonactivated and activated α2β1 integrins in inflammation when compared to thrombosis.

No MeSH data available.


Related in: MedlinePlus

BTT-3016, BTT-3033, and BTT-3034 small molecular inhibitors of α2β1 integrin present anti-inflammatory effect in PAF-induced air pouch model. An air pouch was induced by injecting sterile air to the back of female NMRI mice. Platelet-activating factor (PAF, 0.7 mL of 10−6 mol/L solution/air pouch) was used to induce inflammation. (A) Indicated concentrations of BTT-3016 were administered p.o. and anti-α2 antibody (anti-α2, Ha1/29, 25 μg) directly into air pouch 24 h and 2 h before PAF induction. Number of mice in each group was 10. *P < 0.05; ***P < 0.001 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (B) BTT-3033 and BTT-3034 (both 1 mg/kg, n = 8; 10 mg/kg, n = 8), dexamethasone (Dexam, 0.1 mg/kg, n = 8), nonstimulated vehicle (n = 7) and PAF-stimulated vehicle (n = 8) were administrated p.o. 24 h and 2 h before PAF induction. *P < 0.05; **P < 0.01 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (A and B) The number of leukocytes was counted under microscope. The data are shown as mean ± SEM. (C) The pharmacokinetic profiles of BTT-3033 and BTT-3034 were studied in male DBA/1 mice by administrating single oral dose (10 mg/kg) of the compounds. Plasma concentrations of three animals per sampling time points are presented as mean ± SD. In 1 h time point the difference is statistically significant (P < 0.05; Student’s t-test).
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fig01: BTT-3016, BTT-3033, and BTT-3034 small molecular inhibitors of α2β1 integrin present anti-inflammatory effect in PAF-induced air pouch model. An air pouch was induced by injecting sterile air to the back of female NMRI mice. Platelet-activating factor (PAF, 0.7 mL of 10−6 mol/L solution/air pouch) was used to induce inflammation. (A) Indicated concentrations of BTT-3016 were administered p.o. and anti-α2 antibody (anti-α2, Ha1/29, 25 μg) directly into air pouch 24 h and 2 h before PAF induction. Number of mice in each group was 10. *P < 0.05; ***P < 0.001 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (B) BTT-3033 and BTT-3034 (both 1 mg/kg, n = 8; 10 mg/kg, n = 8), dexamethasone (Dexam, 0.1 mg/kg, n = 8), nonstimulated vehicle (n = 7) and PAF-stimulated vehicle (n = 8) were administrated p.o. 24 h and 2 h before PAF induction. *P < 0.05; **P < 0.01 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (A and B) The number of leukocytes was counted under microscope. The data are shown as mean ± SEM. (C) The pharmacokinetic profiles of BTT-3033 and BTT-3034 were studied in male DBA/1 mice by administrating single oral dose (10 mg/kg) of the compounds. Plasma concentrations of three animals per sampling time points are presented as mean ± SD. In 1 h time point the difference is statistically significant (P < 0.05; Student’s t-test).

Mentions: To analyze the effects of BTT-3016 on acute inflammation, we used a mouse skin air pouch model. This model measures, for example, the function of polymorphonuclear leukocytes, known to use α2β1 as their collagen receptor (Werr et al. 2000). BTT-3016 dose dependently suppressed the recruitment of leukocytes into the air pouch. With 100 mg/kg (oral administration) the effect size was in range of that of 0.1 mg/kg dexamethasone indicating a moderate anti-inflammatory effect (not shown). To further study the dose dependency of BTT-3016 effect we repeated the experiment. Low doses of BTT-3016 (1 and 3 mg/kg, p.o.) were not able to affect the number of leukocytes in inflamed air pouch of mice compared to untreated control group (Fig.1A). However, higher doses (10 and 30 mg/kg) dose dependently and in a statistically significant manner reduced the number of leukocytes after PAF treatment (Fig.1A). Importantly, specific monoclonal antibody against α2 integrin (Ha1/29) was only moderately more effective than BTT-3016. To conclude, our results indicate that orally administrated α2β1 blocking sulfonamides are anti-inflammatory agents in vivo.


Sulfonamide inhibitors of α2β1 integrin reveal the essential role of collagen receptors in in vivo models of inflammation.

Nissinen L, Ojala M, Langen B, Dost R, Pihlavisto M, Käpylä J, Marjamäki A, Heino J - Pharmacol Res Perspect (2015)

BTT-3016, BTT-3033, and BTT-3034 small molecular inhibitors of α2β1 integrin present anti-inflammatory effect in PAF-induced air pouch model. An air pouch was induced by injecting sterile air to the back of female NMRI mice. Platelet-activating factor (PAF, 0.7 mL of 10−6 mol/L solution/air pouch) was used to induce inflammation. (A) Indicated concentrations of BTT-3016 were administered p.o. and anti-α2 antibody (anti-α2, Ha1/29, 25 μg) directly into air pouch 24 h and 2 h before PAF induction. Number of mice in each group was 10. *P < 0.05; ***P < 0.001 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (B) BTT-3033 and BTT-3034 (both 1 mg/kg, n = 8; 10 mg/kg, n = 8), dexamethasone (Dexam, 0.1 mg/kg, n = 8), nonstimulated vehicle (n = 7) and PAF-stimulated vehicle (n = 8) were administrated p.o. 24 h and 2 h before PAF induction. *P < 0.05; **P < 0.01 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (A and B) The number of leukocytes was counted under microscope. The data are shown as mean ± SEM. (C) The pharmacokinetic profiles of BTT-3033 and BTT-3034 were studied in male DBA/1 mice by administrating single oral dose (10 mg/kg) of the compounds. Plasma concentrations of three animals per sampling time points are presented as mean ± SD. In 1 h time point the difference is statistically significant (P < 0.05; Student’s t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4492762&req=5

fig01: BTT-3016, BTT-3033, and BTT-3034 small molecular inhibitors of α2β1 integrin present anti-inflammatory effect in PAF-induced air pouch model. An air pouch was induced by injecting sterile air to the back of female NMRI mice. Platelet-activating factor (PAF, 0.7 mL of 10−6 mol/L solution/air pouch) was used to induce inflammation. (A) Indicated concentrations of BTT-3016 were administered p.o. and anti-α2 antibody (anti-α2, Ha1/29, 25 μg) directly into air pouch 24 h and 2 h before PAF induction. Number of mice in each group was 10. *P < 0.05; ***P < 0.001 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (B) BTT-3033 and BTT-3034 (both 1 mg/kg, n = 8; 10 mg/kg, n = 8), dexamethasone (Dexam, 0.1 mg/kg, n = 8), nonstimulated vehicle (n = 7) and PAF-stimulated vehicle (n = 8) were administrated p.o. 24 h and 2 h before PAF induction. *P < 0.05; **P < 0.01 (One-way ANOVA followed by Bonferroni’s post hoc comparisons tests). (A and B) The number of leukocytes was counted under microscope. The data are shown as mean ± SEM. (C) The pharmacokinetic profiles of BTT-3033 and BTT-3034 were studied in male DBA/1 mice by administrating single oral dose (10 mg/kg) of the compounds. Plasma concentrations of three animals per sampling time points are presented as mean ± SD. In 1 h time point the difference is statistically significant (P < 0.05; Student’s t-test).
Mentions: To analyze the effects of BTT-3016 on acute inflammation, we used a mouse skin air pouch model. This model measures, for example, the function of polymorphonuclear leukocytes, known to use α2β1 as their collagen receptor (Werr et al. 2000). BTT-3016 dose dependently suppressed the recruitment of leukocytes into the air pouch. With 100 mg/kg (oral administration) the effect size was in range of that of 0.1 mg/kg dexamethasone indicating a moderate anti-inflammatory effect (not shown). To further study the dose dependency of BTT-3016 effect we repeated the experiment. Low doses of BTT-3016 (1 and 3 mg/kg, p.o.) were not able to affect the number of leukocytes in inflamed air pouch of mice compared to untreated control group (Fig.1A). However, higher doses (10 and 30 mg/kg) dose dependently and in a statistically significant manner reduced the number of leukocytes after PAF treatment (Fig.1A). Importantly, specific monoclonal antibody against α2 integrin (Ha1/29) was only moderately more effective than BTT-3016. To conclude, our results indicate that orally administrated α2β1 blocking sulfonamides are anti-inflammatory agents in vivo.

Bottom Line: Since α2β1 integrin is abundantly expressed on various inflammation-associated cells, we tested whether recently developed α2β1 blocking sulfonamides have anti-inflammatory properties.Integrin α2β1 inhibitors were shown to reduce the signs of inflammation in arachidonic acid-induced ear edema, PAF stimulated air pouch, ovalbumin-induced skin hypersensitivity, adjuvant arthritis, and collagen-induced arthritis.Thus, the experiments also revealed fundamental differences in the action of nonactivated and activated α2β1 integrins in inflammation when compared to thrombosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, University of Turku 20014, Turku, Finland ; Biotie Therapies Corp Turku, Finland.

ABSTRACT
Small molecule inhibitors of α2β1 integrin, a major cellular collagen receptor, have been reported to inhibit platelet function, kidney injury, and angiogenesis. Since α2β1 integrin is abundantly expressed on various inflammation-associated cells, we tested whether recently developed α2β1 blocking sulfonamides have anti-inflammatory properties. Integrin α2β1 inhibitors were shown to reduce the signs of inflammation in arachidonic acid-induced ear edema, PAF stimulated air pouch, ovalbumin-induced skin hypersensitivity, adjuvant arthritis, and collagen-induced arthritis. Thus, these sulfonamides are potential drugs for acute and allergic inflammation, hypersensitivity, and arthritis. One sulfonamide with potent anti-inflammatory activity has previously been reported to be selective for activated integrins, but not to inhibit platelet function. Thus, the experiments also revealed fundamental differences in the action of nonactivated and activated α2β1 integrins in inflammation when compared to thrombosis.

No MeSH data available.


Related in: MedlinePlus