Targeting L-Proline Uptake as New Strategy for Anti-chagas Drug Development

Abstract

L-Proline is an important amino acid for the pathogenic protists belonging to Trypanosoma and Leishmania genera. In Trypanosoma cruzi, the etiological agent of Chagas disease, this amino acid is involved in fundamental biological processes such as ATP production, differentiation of the insect and intracellular stages, the host cell infection and the resistance to a variety of stresses. In this study, we explore the L-Proline uptake as a chemotherapeutic target for T. cruzi. Novel inhibitors have been proposed containing the amino acid with a linker and a variable region able to block the transporter. A series of sixteen 1,2,3-triazolyl-proline derivatives have been prepared for in vitro screening against T. cruzi epimastigotes and proline uptake assays. We successfully obtained inhibitors that interfere with the amino acid internalization, which validated our design targeting the metabolite's transport. The presented structures are one of few examples of amino acid transporter inhibitors. The unprecedent application of this strategy on the development of new chemotherapy against Chagas disease, opens a new horizon on antiparasitic drug development against parasitic diseases and other pathologies.

Network-Based Approaches Reveal Potential Therapeutic Targets for Host-Directed Antileishmanial Therapy Driving Drug Repurposing

“…Pidolic acid is an active form of 5-oxoproline and occurs in fundamental biological processes such as intracellular stage differentiation, host cell infection, and resistance to various stresses in protozoa belonging to the genera Trypanosoma and Leishmania . Fargnoli et al showed in an unprecedented way that l -proline uptake has been proposed as a chemotherapeutic target for Trypanosoma cruzi , opening a new horizon in the development of new chemotherapeutics against Chagas disease and other parasitic diseases ( 75 ).…”

Proline transport inhibitors trigger differential responses in Trypanosoma cruzi growth inhibition

“…5) was localized to the parasite cytoplasm in clusters resembling intra-organelle localization. The fact that the compound was indeed transported into the cell together with the proline uptake inhibition 22 , allowed us to postulate that: i. compound 1 (from which compound 4 is derived) could be competing with the amino acid incorporation, and ii. compound 1 has a different inhibition mechanism than 3.…”

“…As was mentioned, based on previous work 22,23 , proline analogues 1 -3 (Fig. 1) were selected for this study.…”

“…Considering the important role L-proline has on the energetic metabolism in T. cruzi, and having proved that some of the proline analogues studied here affect the proline uptake and transport in epimastigotes 22,23 , we decided to study how these analogues affect T. cruzi the excretion of end-products of the metabolism. To this end, parasites (0.5x10 6 epimastigotes/mL, strain Dm28c) maintained in DMEM (high glucose concentration) 2% SFB were treated with analogues 1, 2 or 3 at a concentration corresponding to their IC50.…”

“…In a previous work, we have been able to prepare L-proline derivatives as possible inhibitors of the incorporation of this amino acid in T. cruzi. The resulting synthetic library allowed us to find four analogues with IC50 values in CL14 strain T. cruzi epimastigotes under 50 µM 22 , much lower than L-thiazolidine-4-carboxylic acid (T4C, IC50 = 890 µM), the only reported precedent in T. cruzi 14 . Additionally, it was possible to determine that two of these compounds with anti-proliferative activity (1 and 3, Fig.…”