Discovery of the 2,4-Diaminopyrimidine as a Novel Therapeutic Solution for c-Fms and TNF Induced Rheumatoid Arthritis

Overview

Objectives/Goals Rheumatoid arthritis has a worldwide estimated prevalence of 2%, according to the Johns Hopkins Arthritis Center. The current methods to treat this disease involve targeting of the pro-inflammatory cytokine Tumor Necrosis Factor (TNF-alpha) and are not entirely effective. Recent research shows that c-Fms plays a major role in joint deterioration associated with rheumatoid arthritis. My project aimed to discover a novel inhibitor of the TNF cytokine and the c-Fms receptor tyrosine kinase. This inhibitor, 2,4-diaminopyrimidine, could potentially lead to an effective treatment for rheumatoid arthritis. Methods/Materials TNF alpha-expressing Rat2 fibroblast cells were cultured in DMEM+10%FBS+1%Pen-Strep. The fibroblasts were then plated into six well plates for treatment with water (control), Imatinib, and 2,4-diaminopyrimidine. 2,4-Diaminopyrimidine was added at concentrations of 5 uM, 15 uM, and 25 uM, and dilutions were determined based on their molecular weight and calculation from the starting stock, which was 75mM. Imatinib was added at the concentrations of 5 mg/ml, 15 mg/ml, and 25 mg/ml. After a 48 hour incubation, an ELISA assay was conducted to detect TNF levels. TNF alpha concentrations were then measured using a plate reader (spectrophotometer) and converted into pg/ml after plotting the standard curve. Results 2,4-Diaminopyrimidine significantly reduced the concentrations of TNF alpha in a dose dependent manner. In comparison to water, 2,4-diaminopyrimidine at 25 uM reduced TNF alpha levels by 22%, 15 uM reduced TNF alpha levels by 19%, and 5 uM reduced TNF alpha levels by 14%. In comparison to water, Imatinib at 25 mg/ml reduced TNF alpha levels by 10%, 15 mg/ml reduced TNF alpha levels by 9%, and 5 mg/ml reduced TNF alpha levels by 6%. Conclusions/Discussion This research has discovered for the first time that 2,4-diaminopyrimidine inhibits TNF alpha production, supporting my hypothesis. 2,4-Diaminopyrimidine was discovered after investigating into the structures of various inhibitors and analyzing their classifications. In the tests, higher concentrations of 2,4-diaminopyrimidine led to lower levels of TNF alpha. 2,4-Diaminopyrimidine inhibits TNF alpha by binding to the TNF in the cells, preventing its interaction with TNF alpha receptors on the surface of the cells. This discovery could lead to a possible therapeutic solution for c-Fms and TNF alpha induced rheumatoid arthritis.

Awards (1)

• Third Place

Competition history

• CSEF 2011 Pharmacology/ Toxicology · Entry J1724

Resources

• CSEF project summary ↗

Related projects

Closest projects by meaning, across every fair and year in the corpus.

Browse more like this

• All Biomedical and Health Sciences projects
• CSEF Biomedical and Health Sciences projects
• Every CSEF 2011 project
• CSEF Third Place winners