Gold(I) complexes bearing phosphole ligands: Synthesis and antimalarial activity

AbstractA series of gold(I)-monophosphole complexes have been synthesized and characterized. The introduction of a nitrogen moiety in the complex structure was envisioned either by choosing the bis(trifluoromethanesulfonyl)imidate ligand as the X-ligand or by preparing a new pyrrolidinophosphole ligand as the L-ligand. All the complexes have been evaluated in vitro for their antimalarial activity. These gold–phosphole complexes showed moderate activities with IC50 values ranging from 9.7 to 24 μM against Plasmodium falciparum chloroquine-resistant strains

Concentration and purification by magnetic separation of the erythrocytic stages of all human Plasmodium species

International audienceBackground : Parasite concentration methods facilitate molecular, biochemical and immunologicalresearch on the erythrocytic stages of Plasmodium. In this paper, an adaptation of magnetic MACS®columns for the purification of human Plasmodium species is presented. This method was useful forthe concentration/purification of either schizonts or gametocytes.Results and conclusions : The magnetic removal of non-parasitized red blood cells (in vivo andin vitro) using magnetic columns (MACS) was evaluated. This easy-to-use technique enrichedschizonts and gametocytes from Plasmodium falciparum in vitro cultures with a very high degree ofpurity. In addition, all haemozoin-containing stages (schizonts and/or gametocytes) from theperipheral blood of infected patients could be concentrated using this method. This method isparticularly useful for the concentration of non-falciparum species, which do not grow in cultureand are otherwise difficult to obtain in large amounts

Molecular monitoring of plasmodium falciparum drug susceptibility at the time of the introduction of artemisinin-based combination therapy in Yaoundé, Cameroon: Implications for the future

<p>Abstract</p> <p>Background</p> <p>Regular monitoring of the levels of anti-malarial resistance of <it>Plasmodium falciparum </it>is an essential policy to adapt therapy and improve malaria control. This monitoring can be facilitated by using molecular tools, which are easier to implement than the classical determination of the resistance phenotype. In Cameroon, chloroquine (CQ), previously the first-line therapy for uncomplicated malaria was officially withdrawn in 2002 and replaced initially by amodiaquine (AQ) monotherapy. Then, artemisinin-based combination therapy (ACT), notably artesunate-amodiaquine (AS-AQ) or artemether-lumefantrine (AL), was gradually introduced in 2004. This situation raised the question of the evolution of <it>P. falciparum </it>resistance molecular markers in Yaoundé, a highly urbanized Cameroonian city.</p> <p>Methods</p> <p>The genotype of <it>pfcrt </it>72 and 76 and <it>pfmdr1 </it>86 alleles and <it>pfmdr1 </it>copy number were determined using real-time PCR in 447 <it>P. falciparum </it>samples collected between 2005 and 2009.</p> <p>Results</p> <p>This study showed a high prevalence of parasites with mutant <it>pfcrt </it>76 (83%) and <it>pfmdr1 </it>86 (93%) codons. On the contrary, no mutations in the <it>pfcrt </it>72 codon and no samples with duplication of the <it>pfmdr1 </it>gene were observed.</p> <p>Conclusion</p> <p>The high prevalence of mutant <it>pfcrt </it>76T and <it>pfmdr1 </it>86Y alleles might be due to the choice of alternative drugs (AQ and AS-AQ) known to select such genotypes. Mutant <it>pfcrt </it>72 codon was not detected despite the prolonged use of AQ either as monotherapy or combined with artesunate. The absence of <it>pfmdr1 </it>multicopies suggests that AL would still remain efficient. The limited use of mefloquine or the predominance of mutant <it>pfmdr1 </it>86Y codon could explain the lack of <it>pfmdr1 </it>amplification. Indeed, this mutant codon is rarely associated with duplication of <it>pfmdr1 </it>gene. In Cameroon, the changes of therapeutic strategies and the simultaneous use of several formulations of ACT or other anti-malarials that are not officially recommended result in a complex selective pressure, rendering the prediction of the evolution of <it>P. falciparum </it>resistance difficult. This public health problem should lead to increased vigilance and regular monitoring.</p

Ex vivo activity of the ACT new components pyronaridine and piperaquine in comparison with conventional ACT drugs against isolates of Plasmodium falciparum

<p>Abstract</p> <p>Background</p> <p>The aim of the present work was to assess i) <it>ex vivo </it>activity of pyronaridine (PND) and piperaquine (PPQ), as new components of artemisinin-based combination therapy (ACT), to define susceptibility baseline, ii) their activities compared to other partner drugs, namely monodesethylamodiaquine (MDAQ), lumefantrine (LMF), mefloquine (MQ), artesunate (AS) and dihydroartemisinin (DHA) against 181 <it>Plasmodium falciparum </it>isolates from African countries, India and Thailand, and iii) <it>in vitro </it>cross-resistance with other quinoline drugs, chloroquine (CQ) or quinine (QN).</p> <p>Methods</p> <p>The susceptibility of the 181 <it>P. falciparum </it>isolates to the nine anti-malarial drugs was assessed using the standard 42-hours ³H-hypoxanthine uptake inhibition method.</p> <p>Results</p> <p>The IC₅₀values for PND ranged from 0.55 to 80.0 nM (geometric mean = 19.9 nM) and from 11.8 to 217.3 nM for PPQ (geometric mean = 66.8 nM). A significant positive correlation was shown between responses to PPQ and PND responses (<it>rho </it>= 0.46) and between PPQ and MDAQ (<it>rho </it>= 0.30). No significant correlation was shown between PPQ IC₅₀and responses to other anti-malarial drugs. A significant positive correlation was shown between responses to PND and MDAQ (<it>rho </it>= 0.37), PND and LMF (<it>rho </it>= 0.28), PND and QN (<it>rho </it>= 0.24), PND and AS (<it>rho </it>= 0.19), PND and DHA (<it>rho </it>= 0.18) and PND and CQ (<it>rho </it>= 0.16). All these coefficients of correlation are too low to suggest cross-resistance between PPQ or PND and the other drugs.</p> <p>Conclusions</p> <p>In this study, the excellent anti-malarial activity of PPQ and PND was confirmed. The absence of cross-resistance with quinolines and artemisinin derivatives is consistent with the efficacy of the combinations of PPQ and DHA or PND and AS in areas where parasites are resistant to conventional anti-malarial drugs.</p

Nrf2, a PPARγ Alternative Pathway to Promote CD36 Expression on Inflammatory Macrophages: Implication for Malaria

CD36 is the major receptor mediating nonopsonic phagocytosis of Plasmodium falciparum-parasitized erythrocytes by macrophages. Its expression on macrophages is mainly controlled by the nuclear receptor PPARγ. Here, we demonstrate that inflammatory processes negatively regulate CD36 expression on human and murine macrophages, and hence decrease Plasmodium clearance directly favoring the worsening of malaria infection. This CD36 downregulation in inflammatory conditions is associated with a failure in the expression and activation of PPARγ. Interestingly, using siRNA mediating knock down of Nrf2 in macrophages or Nrf2- and PPARγ-deficient macrophages, we establish that in inflammatory conditions, the Nrf2 transcription factor controls CD36 expression independently of PPARγ. In these conditions, Nrf2 activators, but not PPARγ ligands, enhance CD36 expression and CD36-mediated Plasmodium phagocytosis. These results were confirmed in human macrophages and in vivo where only Nrf2 activators improve the outcome of severe malaria. Collectively, this report highlights that the Nrf2 transcription factor could be an alternative target to PPARγ in the control of severe malaria through parasite clearance

Evidence for the Contribution of the Hemozoin Synthesis Pathway of the Murine Plasmodium yoelii to the Resistance to Artemisinin-Related Drugs

Plasmodium falciparum malaria is a major global health problem, causing approximately 780,000 deaths each year. In response to the spreading of P. falciparum drug resistance, WHO recommended in 2001 to use artemisinin derivatives in combination with a partner drug (called ACT) as first-line treatment for uncomplicated falciparum malaria, and most malaria-endemic countries have since changed their treatment policies accordingly. Currently, ACT are often the last treatments that can effectively and rapidly cure P. falciparum infections permitting to significantly decrease the mortality and the morbidity due to malaria. However, alarming signs of emerging resistance to artemisinin derivatives along the Thai-Cambodian border are of major concern. Through long-term in vivo pressures, we have been able to select a murine malaria model resistant to artemisinins. We demonstrated that the resistance of Plasmodium to artemisinin-based compounds depends on alterations of heme metabolism and on a loss of hemozoin formation linked to the down-expression of the recently identified Heme Detoxification Protein (HDP). These artemisinins resistant strains could be able to detoxify the free heme by an alternative catabolism pathway involving glutathione (GSH)-mediation. Finally, we confirmed that artemisinins act also like quinolines against Plasmodium via hemozoin production inhibition. The work proposed here described the mechanism of action of this class of molecules and the resistance to artemisinins of this model. These results should help both to reinforce the artemisinins activity and avoid emergence and spread of endoperoxides resistance by focusing in adequate drug partners design. Such considerations appear crucial in the current context of early artemisinin resistance in Asia

Do ethnobotanical and laboratory data predict clinical safety and efficacy of anti-malarial plants?

<p>Abstract</p> <p>Background</p> <p>Over 1200 plant species are reported in ethnobotanical studies for the treatment of malaria and fevers, so it is important to prioritize plants for further development of anti-malarials.</p> <p>Methods</p> <p>The “RITAM score” was designed to combine information from systematic literature searches of published ethnobotanical studies and laboratory pharmacological studies of efficacy and safety, in order to prioritize plants for further research. It was evaluated by correlating it with the results of clinical trials.</p> <p>Results and discussion</p> <p>The laboratory efficacy score correlated with clinical parasite clearance (r_s=0.7). The ethnobotanical component correlated weakly with clinical symptom clearance but not with parasite clearance. The safety component was difficult to validate as all plants entering clinical trials were generally considered safe, so there was no clinical data on toxic plants.</p> <p>Conclusion</p> <p>The RITAM score (especially the efficacy and safety components) can be used as part of the selection process for prioritising plants for further research as anti-malarial drug candidates. The validation in this study was limited by the very small number of available clinical studies, and the heterogeneity of patients included.</p

Next-Generation Antimalarial Drugs: Hybrid Molecules as a New Strategy in Drug Design

Malaria is a disease that affects nearly 40% of the global population, and chemotherapy remains the mainstay of its control strategy. The global malaria situation is increasingly being exacerbated by the emergence of drug resistance to most of the available antimalarials, necessitating search for novel drugs. A recent rational approach of antimalarial drug design characterized as “covalent bitherapy” involves linking two molecules with individual intrinsic activity into a single agent, thus packaging dual-activity into a single hybrid molecule. Current research in this field seems to endorse hybrid molecules as the next-generation antimalarial drugs. If the selective toxicity of hybrid prodrugs can be demonstrated in vivo with good bioavailability at the target site in the parasite, it would offer various advantages including dosage compliance, minimized toxicity, ability to design better drug combinations, and cheaper preclinical evaluation while achieving the ultimate object of delaying or circumventing the development of resistance. This review is focused on several hybrid molecules that have been developed, with particular emphasis on those deemed to have high potential for development for clinical use. Drug Dev Res 71: 20–32, 2010. © 2009 Wiley-Liss, Inc