<i>In Vitro</i>and<i>In Vivo</i>Antiplasmodial Activities of Risedronate and Its Interference with Protein Prenylation in Plasmodium falciparum

Jordão, Fabiana Morandi; Saito, Alexandre Yukio; Miguel, Danilo C.; Peres, Valnice de Jesus; Kimura, Emı́lia A.; Katzin, Alejandro M.

doi:10.1128/aac.01820-10

Cited by 34 publications

(36 citation statements)

References 43 publications

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“…Once IPP and DMAPP are synthesized and transported out of the apicoplast, a myriad of isoprenoid products with different cellular localization and functions are synthesized in the malaria parasite. Many isoprenoid products have been identified as being present in the intraerythrocytic asexual stages of P. falciparum, such as ubiquinones and menaquinone (32,33,56), carotenoids and vitamin E (57-59), dolichols (50,60,61), and prenylated proteins (60,(62)(63)(64)(65). The presence and function of these isoprenoids in gametocytes remain to be confirmed and are under investigation.…”

Section: Discussionmentioning

confidence: 99%

Isoprenoid Precursor Biosynthesis Is the Essential Metabolic Role of the Apicoplast during Gametocytogenesis in Plasmodium falciparum

et al. 2015

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The malaria parasite harbors a relict plastid called the apicoplast and its discovery opened a new avenue for drug discovery and development due to its unusual, nonmammalian metabolism. The apicoplast is essential during the asexual intraerythrocytic and hepatic stages of the parasite, and there is strong evidence supporting its essential metabolic role during the mosquito stages of the parasite. Supply of the isoprenoid building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) is the essential metabolic function of the apicoplast during the asexual intraerythrocytic stages. However, the metabolic role of the apicoplast during gametocyte development, the malaria stages transmitted to the mosquito, remains unknown. In this study, we showed that production of IPP for isoprenoid biosynthesis is the essential metabolic function of the apicoplast during gametocytogenesis, by obtaining normal gametocytes lacking the apicoplast when supplemented with IPP. When IPP supplementation was removed early in gametocytogenesis, developmental defects were observed, supporting the essential role of isoprenoids for normal gametocytogenesis. Furthermore, mosquitoes infected with gametocytes lacking the apicoplast developed fewer and smaller oocysts that failed to produce sporozoites. This finding further supports the essential role of the apicoplast in establishing a successful infection in the mosquito vector. Our study supports isoprenoid biosynthesis as a valid drug target for development of malaria transmission-blocking inhibitors.

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Section: Discussionmentioning

confidence: 99%

Isoprenoid Precursor Biosynthesis Is the Essential Metabolic Role of the Apicoplast during Gametocytogenesis in Plasmodium falciparum

et al. 2015

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“…BPH-811 is an analog of risedronate in which, again, the 1-OH group is removed and a lipophilic tail added. Both compounds have potent activity in vitro and in vivo, whereas the parent drugs, zoledronate and risedronate, have only modest activity against P. falciparum in intraerythrocytic assays; and risedronate, although reducing parasitaemia, has no effect on survival (15). Also of interest is the observation that the lipophilic bisphosphonate inhibitors in the Plasmodium GGPPS structures adopt very similar binding poses to those of their parent compounds bound to both GGPPS as well as FPPS, and have similar enzyme activity and ΔG values, for GGPPS binding.…”

Section: Discussionmentioning

confidence: 99%

“…(4,7,8), Toxoplasma gondii (4, 9), Cryptosporidium parvum (10,11), Entamoeba histolytica (4,12,13), and Plasmodium spp. (4,(13)(14)(15). In the case of Plasmodium spp., the most potent inhibitors were not, however, the nitrogencontaining bisphosphonates such as zoledronate or risedronate (Scheme 1) used to treat bone diseases, but more lipophilic n-alkyl bisphosphonates (13).…”

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confidence: 99%

Lipophilic analogs of zoledronate and risedronate inhibit Plasmodium geranylgeranyl diphosphate synthase (GGPPS) and exhibit potent antimalarial activity

Dossin

Zhang

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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We report the results of an in vitro screening assay targeting the intraerythrocytic form of the malaria parasite Plasmodium falciparum using a library of 560 prenyl-synthase inhibitors. Based on "growth-rescue" and enzyme-inhibition experiments, geranylgeranyl diphosphate synthase (GGPPS) is shown to be a major target for the most potent leads, BPH-703 and BPH-811, lipophilic analogs of the bone-resorption drugs zoledronate and risedronate. We determined the crystal structures of these inhibitors bound to a Plasmodium GGPPS finding that their head groups bind to the ½Mg 2þ 3 cluster in the active site in a similar manner to that found with their more hydrophilic parents, whereas their hydrophobic tails occupy a long-hydrophobic tunnel spanning both molecules in the dimer. The results of isothermal-titration-calorimetric experiments show that both lipophilic bisphosphonates bind to GGPPS with, on average, a ΔG of −9 kcal mol −1 , only 0.5 kcal mol −1 worse than the parent bisphosphonates, consistent with the observation that conversion to the lipophilic species has only a minor effect on enzyme activity. However, only the lipophilic species are active in cells. We also tested both compounds in mice, finding major decreases in parasitemia and 100% survival. These results are of broad general interest because they indicate that it may be possible to overcome barriers to cell penetration of existing bisphosphonate drugs in this and other systems by simple covalent modification to form lipophilic analogs that retain their enzyme-inhibition activity and are also effective in vitro and in vivo.M alaria, caused by Plasmodium spp., causes approximately 1 million deaths each year (1), and there are ever-present problems due to drug resistance (2). There is, therefore, a need for new drugs and drug leads. In earlier work, we and others found that the bisphosphonate class of drugs (3) used to treat bonerelated diseases-osteoporosis, Paget disease, and hypercalcemia due to malignancy-also inhibited the growth of a range of parasitic protozoa, including Trypanosoma cruzi (4, 5), Trypanosoma brucei (4, 6), Leishmania spp. (4,7,8), Toxoplasma gondii (4, 9), Cryptosporidium parvum (10, 11), Entamoeba histolytica (4, 12, 13), and Plasmodium spp. (4, 13-15). In the case of Plasmodium spp., the most potent inhibitors were not, however, the nitrogencontaining bisphosphonates such as zoledronate or risedronate (Scheme 1) used to treat bone diseases, but more lipophilic n-alkyl bisphosphonates (13). Their target in Plasmodium falciparum was not determined. However, more recently, a Plasmodium vivax geranylgeranyl diphosphate synthase (PvGGPPS) has been cloned, expressed, purified, and crystallized, and its three-dimensional structure determined (16). The enzyme is inhibited by bisphosphonates (16), so it seemed possible that it might be a target for the inhibitors discovered earlier. To investigate this possibility, we recently determined the IC 50 values for 25 bisphosphonates against PvGGPPS and compared the results for enzyme inhi...

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“…We evaluated the combination of four drugs, three of them (fosmidomycin, nerolidol and risedronate) inhibiting parasite growth at some point in the MEP/isoprenoid pathway in intraerythrocytic stages of P. falciparum (6,7,8). We also evaluated the effect of squalestatin on P. falciparum growth.…”

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confidence: 99%

In Vitro Antimalarial Activity of Different Inhibitors of the Plasmodial Isoprenoid Synthesis Pathway

Silva

Saito

Peres

et al. 2015

Antimicrob Agents Chemother

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Previous studies have shown that fosmidomycin, risedronate, and nerolidol exert antimalarial activity in vitro. We included squalestatin, an inhibitor of the isoprenoid metabolism in Erwinia uredovora, and found that combinations of compounds which act on different targets of the plasmodial isoprenoid pathway possess important supra-additivity effects.M alaria expansion in some areas has been attributed to the failure of vector-control policies and, mainly, to the increase of parasite resistance to drugs commonly used for its therapy (1). This alarming scenario has fuelled research in new antimalarial drugs, and, with the sequencing of parasite genomes, new potential drug targets have emerged. Burrows et al. published a review discussing the strategies for designing the next generation of medicines for malaria control (2). A metabolic pathway that may be a likely target for evaluating potential antimalarials is the isoprenoid pathway. The first intermediates are biosynthesized in the apicoplast, which is an organelle present in protozoan parasites of the subphylum Apicomplexa, including Plasmodia (3).We have previously demonstrated that the isoprenoid biosynthesis pathway is functionally active in the intraerythrocytic stages of Plasmodium falciparum and that final products of this pathway such as dolichol of 11 to 12 isoprenic units (4), ubiquinones (5), isoprenylated/dolichylated proteins, carotenoids, menaquinone, and tocopherol are biosynthesized by this parasite (6). Several inhibitors of enzymes of the methyl erythritol diphosphate (MEP) and isoprenoid pathways were described as potential antimalarial drugs (6).Our aim in this work was to identify and validate different targets in the isoprenoid pathway in P. falciparum intraerythrocytic stages. We evaluated the combination of four drugs, three of them (fosmidomycin, nerolidol and risedronate) inhibiting parasite growth at some point in the MEP/isoprenoid pathway in intraerythrocytic stages of P. falciparum (6,7,8). We also evaluated the effect of squalestatin on P. falciparum growth. This drug is an inhibitor of the phytoene synthase in Erwinia uredovora (9, 10). Importantly, an enzyme with phytoene synthase activity was described in intraerythrocytic stages of P. falciparum (11).Parasites of P. falciparum clone 3D7 were cultured according to the protocol described by Trager and Jensen (12) where human serum was replaced by Albumax I (0.5%) (4). Stocks (1 mM) of squalestatin, risedronate, and fosmidomycin were prepared in RPMI medium, whereas nerolidol was diluted in ethanol, and all drugs were stored at Ϫ70°C.Assays were performed in 96-well microtiter plates containing 100 l of 3D7 culture at 1% parasitemia and 3% hematocrit and 100 l of the appropriate drug or drug combination in RPMI medium. Parasite growth was determined using Giemsa-stained smears immediately before the start of the assay and at intervals of 24 to 48 h. Vehicle controls were included on each plate. All tests were performed in triplicate for three independent experiments.Values for 5...

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In VitroandIn VivoAntiplasmodial Activities of Risedronate and Its Interference with Protein Prenylation in Plasmodium falciparum

Cited by 34 publications

References 43 publications

Isoprenoid Precursor Biosynthesis Is the Essential Metabolic Role of the Apicoplast during Gametocytogenesis in Plasmodium falciparum

Isoprenoid Precursor Biosynthesis Is the Essential Metabolic Role of the Apicoplast during Gametocytogenesis in Plasmodium falciparum

Lipophilic analogs of zoledronate and risedronate inhibit Plasmodium geranylgeranyl diphosphate synthase (GGPPS) and exhibit potent antimalarial activity

In Vitro Antimalarial Activity of Different Inhibitors of the Plasmodial Isoprenoid Synthesis Pathway

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