The online version of this article (doi:10.1186/s13075-017-1236-x) contains supplementary material, which is available to authorized users.
The inhibition of pyrimidine biosynthesis by blocking the dihydroorotate dehydrogenase (DHODH) activity, the prime target of leflunomide (LEF), has been proven to be an effective strategy for rheumatoid arthritis (RA) treatment. However, a considerable proportion of RA patients are refractory to LEF. Here, we investigated lapachol (LAP), a natural naphthoquinone, as a potential DHODH inhibitor and addressed its immunosuppressive properties.
Molecular flexible docking studies and bioactivity assays were performed to determine the ability of LAP to interact and inhibit DHODH. In vitro studies were conducted to assess the antiproliferative effect of LAP using isolated lymphocytes. Finally, collagen-induced arthritis (CIA) and antigen-induced arthritis (AIA) models were employed to address the anti-arthritic effects of LAP.
We found that LAP is a potent DHODH inhibitor which had a remarkable ability to inhibit both human and murine lymphocyte proliferation in vitro. Importantly, uridine supplementation abrogated the antiproliferative effect of LAP, supporting that the pyrimidine metabolic pathway is the target of LAP. In vivo, LAP treatment markedly reduced CIA and AIA progression as evidenced by the reduction in clinical score, articular tissue damage, and inflammation.
Our findings propose a binding model of interaction and support the ability of LAP to inhibit DHODH, decreasing lymphocyte proliferation and attenuating the severity of experimental autoimmune arthritis. Therefore, LAP could be considered as a potential immunosuppressive lead candidate with potential therapeutic implications for RA.
Additional file 1: Figure S1. Mean concentration–time profiles of lapachol after (A) 2 mg/kg i.v., (B) 10 mg/kg oral, and (C) 25 mg/kg oral administration to rats. Data points are mean ± standard deviation. (PDF 146 kb)
Additional file 2: TableS1. LAP and LAP sodium salt pharmacokinetic parameters determined by noncompartmental and compartmental approaches after i.v. administration of 2 mg/kg in Wistar rats. (PDF 93 kb)
Additional file 3: Table S2. Pharmacokinetic parameters after oral administration of lapachol (10 mg/kg and 25 mg/kg) and LAP sodium salt (30 mg/kg) in Wistar rats. (PDF 79 kb)
Additional file 4: Figure S2. Superimposition of the crystallographic hDHODH inhibitor A771726 (PDB id:1D3H, carbon atoms in cyan) and the top-ranked docking solution (carbon atoms in yellow), inside the active site. (PDF 458 kb)
Additional file 5: Table S3. Evaluation of the toxicity of lapachol in proliferating CD4 T cells. (PDF 224 kb)
Additional file 6: Figure S3. Serum levels of GPT and AST in LAP-treated mice during CIA protocol. (PDF 518 kb)
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- Lapachol, a compound targeting pyrimidine metabolism, ameliorates experimental autoimmune arthritis
Raphael S. Peres
Gabriela B. Santos
Nerry T. Cecilio
Valquíria A. P. Jabor
Bruna G. S. Torres
Carlos H. T. P. Silva
Teresa Dalla Costa
Norberto P. Lopes
Maria C. Nonato
Fernando S. Ramalho
Thiago M. Cunha
Fernando Q. Cunha
Flavio S. Emery
Jose C. Alves-Filho
- BioMed Central
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