Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

In vitro anti-leishmania activity and safety of newly synthesized thiazolo pyrimidine derivatives augmented with interleukine-12 (IL-12) in BALB/c mice experimentally-infected with cutaneous leishmaniasis

Saleh A Bahashwan¹ , Mohamed A Ramadan², Moutasem S Aboonq³, Ahmed A Fayed⁴

¹Pharmacology and Toxicology Department, College of Pharmacy; ²Microbiology/Immunology Department; ³Physiology Department, College of Medicine; ⁴Respiratory Therapy Department, College of Medical Rehabilitation Sciences, Taibah University, Medina Munawarah, Saudi Arabia.

For correspondence:- Saleh Bahashwan Email: bhswn@yahoo.com Tel:+966-590045053

Received: 9 May 2015 Accepted: 7 October 2015 Published: 29 November 2015

Citation: Bahashwan SA, Ramadan MA, Aboonq MS, Fayed AA. In vitro anti-leishmania activity and safety of newly synthesized thiazolo pyrimidine derivatives augmented with interleukine-12 (IL-12) in BALB/c mice experimentally-infected with cutaneous leishmaniasis. Trop J Pharm Res 2015; 14(11):1975-1981 doi: 10.4314/tjpr.v14i10.4

© 2015 The authors.
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Abstract

Purpose: To synthesize a series of novel thiazolo pyrimidine derivatives and evaluate them in vitro and in vivo for their safety and anti-leishmanial activity using BALB/c mice.

Methods: Substituted pyrazolopyrimidine and pyrazolopyrazole were synthesized by reacting amino group of 2-amino-4-cyano-pyrazol]naphthalino[1,2-d]thiazole with a variety of formamide or hydrazine hydrate. The synthesized compounds were characterized by nuclear magnetic resonance spectroscopy (¹H-NMR) and mass spectroscopy (MS). The purity of the compounds was determined by elemental analysis. Safety and anti-leishmanial activity of the compounds were determined in vitro by i) viability assessment of leishmania-infected macrophages, relative abundance of IL-12p40 mRNA gene expression and levels of IL10 /IL-12 determination in supernatants of cultured macrophages treated with 2.5 and 10 µM of the compounds, using microscope cell counting, reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA), respectively. ii) cytotoxicity of the compounds evaluated by determination the safety index as IC₅₀ of the compound in macrophages/IC₅₀ of the compound in amastigotes. iii) bioassay at 16 weeks post-infection of mice treated with the reference drug, the tested compound alone and both the compound with IL-12. Disease progression and footpad thickness were evaluated regularly during treatment.

Results: Compound 4 emerged as the most active anti-protozoal compound of the series against Leishmania viability (activity 60 %) compared with the reference drug (activity 65 %). When it was combined with IL-12, the activity reached 90 %.

Conclusion: Compound 4 can serve as a lead molecule for further development to a clinically useful novel class of agents.