Malononitrilamides 715 and 279 prevent accelerated cardiac allograft rejection synergistically with cyclosporin A in presensitized rats.

A77 1726 is a malononitrilarnide (MNA) and the active metabolite of leflunomide. Leflunomide has been extensively investigated and shown to be a potent immunosuppressive drug. However, the half-life of A77 1726 is about 15-18 days in humans and the leflunomide is, therefore, currently being evaluated for the treatment of autoimmune disease and not for transplantation. The search for analogues has led to the development of MNA 715 and 279, derivatives of A77 1726. Previous limited experimental experience has shown these MNAs to prevent skin allograft and xenograft rejection and graft-versus-host disease in rodents, and to reverse ongoing allograft rejection. The aim of the present study was to verify the efficacy of these MNAs in a cardiac retransplant model of sensitized rats, concerning prevention of accelerated rejection, inhibition of antibody production and interaction with cyclosporin A (CsA). Heterotopic cardiac transplantation and retransplantation in Dark Agouti (DA) to Piebald Virol Glaxo (PVG) rats was used. Subgroups of rats were given either CsA, MNA 715, MNA 279 or combined CsA/MNA for 10 days starting either day 0 or day -1, as of regrafting or no treatment. Titres of allospecific IgM and IgG were quantitated by flow cytometry. Ten days of MNA 715 or 279 from day -1 prevented accelerated rejection of the retransplant, as did CsA. Neither treatment given from day 0 prevented rejection within 24 h. However, a combination of MNA 715 or 279 and CsA from day 0 effectively prevented accelerated regraft rejection. Production of specific alloantibodies was reduced in all immunosuppressed subgroups, IgG titres at day 7 in MNA-treated subgroups being significantly lower compared with those in the CsA-treated subgroup. In conclusion, MNA 715 and 279 were shown to be potent immunosuppressants with the capacity to prevent accelerated regraft rejection in rat cardiac transplants, most efficiently in combination with CsA, and to suppress specific alloantibody production.