Role of angiotensin pathway and its target therapy to rescue from experimental cerebral malaria.

Cerebral malaria (CM) induced by Plasmodium falciparum is a devastating neurological complication that may lead the patient to coma and death. This study aimed to protect Plasmodium-infected C57BL6 mice from CM by targeting the angiotensin II type 1 (AT1) receptor, which is considered the common connecting link between hypertension and CM. In CM, AT-1 mediates blood-brain barrier (BBB) damage through the overexpression of ß-catenin. The AT-1-inhibiting drugs, such as irbesartan and losartan, were evaluated for the prevention of CM. The effectiveness of these drugs was determined by the down regulation of ß-catenin, TCF, LEF, ICAM-1, and VCAM-1 in the drug-treated groups. The expression levels of VE-cadherin and vinculin, essential for the maintenance of BBB integrity, were found to be restored in the drug-treated groups. The pro-inflammatory cytokine levels were decreased, and the anti-inflammatory cytokine levels increased with the treatment. As a major highlight, the mean survival time of treated mice was found to be increased even in the absence of treatment with an anti-malarial agent. The combination of irbesartan or losartan with the anti-malarial agent α/ß-arteether has contributed to an 80% cure rate, which is higher than the 60% cure rate observed with α/ß-arteether alone treatment.

Metacaspase (Pf MCA-1) as antimalarial drug target: An in silico approach and their biological validation.

AIMS: Acquired drug resistance of Plasmodium is a global issue for the treatment of malaria. There are various proteases in the genome of Plasmodium falciparum (P. falciparum) including metacaspase-1 (PfMCA-1) that are essential and are being considered as an attractive drug target. It is aimed to identify novel therapeutics against malaria and their action on PfMCA-1 along with other apoptotic pathway events. MAIN METHODS: High throughput virtual screening of 55,000 compounds derived from Maybridge library was performed against PfMCA-1. Based on the docking score, sixteen compounds were selected for in vitro antimalarial screening against drug sensitive and resistant strains of P. falciparum using SYBR green-based assay. Subsequently, three lead molecules were selected and subjected to the evaluation of cytotoxicity, caspase like protease activity, mitochondrial membrane potential, ROS generation and DNA fragmentation via TUNEL assay. KEY FINDINGS: The in silico and in vitro approaches have brought forward some Maybridge library compounds with antiplasmodial activity most likely by enhancing the metacaspase activity. The compound CD11095 has shown better antimalarial efficacy, and KM06591 depicted higher caspase mediated killing, elevated TUNEL positive cells and moderate ROS generation. Mitochondrial membrane depolarization was augmented by RJC0069. Exposure of P. falciparum to CD11095, KM06591 and RJC0069 has ended up in parasite growth arrest via multiple mechanisms. SIGNIFICANCE: It is proposed that the Maybridge molecules CD11095, KM06591 and RJC0069 have antimalarial activity. Their mechanism of action was found to be by enhancing the metacaspases-like protease activity, mitochondrial depolarization and DNA fragmentation which stipulates significant insights towards promising candidates for drug development.

Molecular and immunological characterization of the calcyclin binding protein in rodent malaria parasite.

Malaria remains as a global life-threatening disorder due to the emergence of resistance against standard antimalarials. Consequently, there is a serious need to better understand the biology of the malaria parasite in order to determine appropriate targets for new interventions. Calcyclin binding protein (CacyBP) is a multi-functional and multi-ligand protein that is not well characterized in malaria disease. In this study, we have cloned CacyBP from rodent species Plasmodium yoelii nigeriensis and purified the recombinant protein to carry out its detailed molecular, biophysical and immunological characterization. Molecular characterization indicates that PyCacyBP is a ∼27 kDa protein in parasite lysate and exists in monomer and dimer forms. Bioinformatic analysis of CacyBP showed significant sequence and structural similarities between rodent and human malaria parasites. CacyBP is expressed in all blood stages of P. yoelii nigeriensis parasite. In silico studies proposed the immunogenic potential of CacyBP. The rPyCacyBP immunized mice exhibited elevated levels of IgG1, IgG2a, IgG2b and IgG3 in their serum. Notably, cellular immune response in splenocytes from immunized mice showed increased expression of pro-inflammatory cytokines such as IL-12, IFN-γ and TNF-α. This CacyBP exhibited pro-inflammatory immune response in rodent host. These finding revealed that CacyBP may have the potential to boost the host immunity for protection against malaria infection. The present study provides basis for further exploration of the biological function of CacyBP in malaria parasite.

Structural characterization of Kannurin isoforms and evaluation of the role of ß-hydroxy fatty acid tail length in functional specificity.

The novel anti-fungal cyclic lipopeptide 'Kannurin' and its three structural variants produced by Bacillus cereus AK1 were previously reported from our laboratory. The present study reports unexplored structural variants of Kannurin those have functional benefits. Due to the difference in ß-hydroxy fatty acid tail length, they are designated here as Kannurin A (m/z 994.67 ± 0.015), B (m/z 1008.68 ± 0.017), C (m/z 1022.69 ± 0.021), D (m/z 1036.70 ± 0.01), CL (m/z 1040.71 ± 0.02) and DL (m/z 1054.72 ± 0.01). The isoform A (m/z 994.67 ± 0.015) is the shortest cyclic form of Kannurin identified so far. In addition, CL (m/z 1040.71 ± 0.02) and DL (m/z 1054.72 ± 0.01) are the rare natural linear forms. The results of the antimicrobial assays deduced that the difference in lipid tail length of the isoforms contributes tremendous differences in their antimicrobial properties. The isoforms with short lipid tails (A and B) are more selective and potent towards bacteria, whereas the isoforms with long lipid tails (C and D) are more potent against fungi. The molecular dynamics studies and electron microscopic observations supported with circular dichroic spectroscopy analysis showed the structural confirmation and formation of aggregates of Kannurin in solution. The molecular dynamics simulation studies revealed that a single molecule of Kannurin makes enormous intra-molecular interactions and structural re-arrangements to attain stable lowest energy state in solution. When they reach a particular concentration (CMC) especially in aqueous environment, tends to form structural aggregates called 'micelles'. With the structural information and activity relationship described in this study, it is trying to point out the sensitive structural entities that can be modified to improve the efficacy and target specificities of lipopeptide class of antibiotics.