ABSTRACT
We have reported that two endoperoxides, N-89 and N-251, synthesized in 2001, possess potent antimalarial activities. Aiming at their eventual use for curing malaria in humans, we have been investigating various aspects of their antimalarial actions. Here we show that N-89 and N-251 inhibit the growth of Plasmodium falciparum within human erythrocytes in vitro at its lifecycle stage 'trophozoite' specifically. It is known that artemisinin compounds, which are currently used for curing malaria, have other stage-specificities. Therefore, it is likely that the antimalarial mechanism of N-89 and N-251 differs from those of artemisinin compounds. As malaria parasites resistant to artemisinin-based combination therapy are currently emerging in some tropical regions, N-89 and N-251 are candidates for overcoming these new problems.
Subject(s)
Antimalarials/pharmacology , Heterocyclic Compounds, 2-Ring/pharmacology , Plasmodium falciparum/drug effects , Spiro Compounds/pharmacology , Tetraoxanes/pharmacology , Animals , Antimalarials/chemical synthesis , Artemisinins/pharmacology , Drug Resistance, Multiple , Erythrocytes/parasitology , Humans , Malaria/parasitology , Parasitic Sensitivity Tests , Plasmodium falciparum/growth & development , Trophozoites/drug effects , Trophozoites/ultrastructureABSTRACT
The endoperoxide artemisinin is a current first-line antimalarial and a critical component of the artemisinin-based combination therapies (ACT) recommended by WHO for treatment of Plasmodium falciparum, the deadliest of malaria parasites. However, recent emergence of the artemisinin-resistant P. falciparum urged us to develop new antimalarial drugs. We have shown that synthetic endoperoxides N-89 and its hydroxyl derivative N-251 had high antimalarial activities both in vivo and in vitro. However, the mechanisms including the cellular targets of the endoperoxide antimalarials are not well understood. Thus, in this study, we employed chemical proteomics to survey potential molecular targets of endoperoxides by evaluating P. falciparum proteins capable to associate with endoperoxide structure (N-346, a carboxyamino derivative of N-89). We also analyzed the protein expression profiles of malaria parasites treated with N-89 or N-251 to explore possible changes associated with the drug action. From these experiments, we found that P. falciparum endoplasmic reticulum-resident calcium binding protein (PfERC) had high affinity to the endoperoxide structure (N-346) and was decreased by treatment with N-89 or N-251. PfERC is a member of CREC protein family, a potential disease marker and also a potential target for therapeutic intervention. We propose that the PfERC is a strong candidate of the endoperoxide antimalarial's target.
Subject(s)
Antimalarials/pharmacology , Calcium-Binding Proteins/chemistry , Endoplasmic Reticulum/chemistry , Peroxides/pharmacology , Plasmodium falciparum/chemistry , Protozoan Proteins/chemistry , Antimalarials/chemistry , Chromatography, Affinity , Electrophoresis, Polyacrylamide Gel , Erythrocytes/parasitology , Heterocyclic Compounds, 2-Ring/pharmacology , Humans , Peroxides/chemistry , Plasmodium falciparum/drug effects , Plasmodium falciparum/pathogenicity , Proteomics/methods , Recombinant Proteins/chemistry , Spiro Compounds/pharmacology , Tetraoxanes/pharmacology , Trophozoites/chemistry , Trophozoites/drug effectsABSTRACT
Malaria is one of the world's deadliest diseases and is becoming an increasingly serious problem as malaria parasites develop resistance to most of the antimalarial drugs used today. We previously reported the in vitro and in vivo antimalarial potencies of 1,2,6,7-tetraoxaspiro[7.11]nonadecane (N-89) and 6-(1,2,6,7-tetraoxaspiro[7.11]nonadec-4-yl)hexan-1-ol (N-251) against Plasmodium falciparum and Plasmodium berghei parasites. To improve water-solubility for synthetic peroxides, a variety of cyclic peroxides having carboxyl functionality was prepared based on the antimalarial candidate, N-251, and their antimalarial activities were determined. The reactions of N-89 and its derivatives with Fe(II) demonstrated a highly efficient formation of the corresponding carbon radical which may be suspected as a key for the antiparasitic activity.
Subject(s)
Antimalarials/administration & dosage , Hexanols/administration & dosage , Malaria, Falciparum/drug therapy , Malaria/drug therapy , Plasmodium berghei/drug effects , Plasmodium falciparum/drug effects , Spiro Compounds/administration & dosage , Animals , Antimalarials/chemical synthesis , Antimalarials/therapeutic use , Carbon/chemistry , Carbon/metabolism , Carboxylic Acids/chemistry , Drug Evaluation, Preclinical , Ferrous Compounds/metabolism , Free Radicals/chemistry , Free Radicals/metabolism , Hexanols/chemical synthesis , Hexanols/therapeutic use , Humans , Inhibitory Concentration 50 , Malaria/parasitology , Malaria, Falciparum/parasitology , Mice , Mice, Inbred ICR , Oxidation-Reduction , Peroxides/chemistry , Peroxides/metabolism , Plasmodium berghei/growth & development , Plasmodium falciparum/growth & development , Spiro Compounds/chemical synthesis , Spiro Compounds/therapeutic use , Structure-Activity RelationshipABSTRACT
Plasmodium falciparum, the major causative parasite for the disease, has acquired resistance to most of the antimalarial drugs used today, presenting an immediate need for new antimalarial drugs. Here, we report the in vitro and in vivo antimalarial activities of 6-(1,2,6,7-tetraoxaspiro[7.11]nonadec-4-yl)hexan-1-ol (N-251) against P. falciparum and Plasmodium berghei parasites. The N-251 showed high antimalarial potencies both in the in vitro and the in vivo tests (EC(50) 2.3×10(-8) M; ED(50) 15 mg/kg (per oral)). The potencies were similar to that of artemisinin in vitro and greater than artemisinin's activity in vivo (p.o.). In addition, N-251 has little toxicity: a single oral administration at 2000 mg/kg to a rat gave no health problems to it. Administration of N-251 to mice bearing 1% of parasitemia (per oral 68 mg/kg, 3 times a day for 3 consecutive days) resulted in a dramatic decrease in the parasitemia: all the 5 mice given N-251 were cured without any recurrence, with no diarrhea or weight loss occurring in the 60 days of experiment. N-251 deserves more extensive clinical evaluation, desirably including future trials in the human.
Subject(s)
Antimalarials/pharmacology , Artemisinins/pharmacology , Heterocyclic Compounds, 2-Ring/pharmacology , Hexanols/pharmacology , Malaria, Falciparum/drug therapy , Plasmodium berghei/drug effects , Plasmodium falciparum/drug effects , Spiro Compounds/pharmacology , Administration, Oral , Animals , Antimalarials/chemistry , Antimalarials/therapeutic use , Artemisinins/therapeutic use , Cell Line, Tumor , Drug Therapy, Combination , Erythrocytes/drug effects , Erythrocytes/parasitology , Heterocyclic Compounds, 2-Ring/chemical synthesis , Heterocyclic Compounds, 2-Ring/therapeutic use , Hexanols/chemistry , Hexanols/therapeutic use , Humans , Malaria/drug therapy , Malaria/parasitology , Malaria, Falciparum/parasitology , Mice , Mice, Inbred ICR , Molecular Structure , Parasitemia/drug therapy , Parasitic Sensitivity Tests , Rats , Spiro Compounds/chemical synthesis , Spiro Compounds/chemistry , Spiro Compounds/therapeutic use , Survival Analysis , TetraoxanesABSTRACT
1,2,6,7-Tetraoxaspiro[7.11]nonadecane (N-89) is a chemically synthesized compound with good efficacy against malaria parasites. We observed strong anti-schistosomal activities of N-89 both in vitro and in vivo. In a murine model with experimental infection of Schistosoma mansoni, orally administered N-89 at the dose of 300 mg/kg resulted in a significant reduction in worm burden (63%) when mice were treated at 2-weeks postinfection. Strong larvicidal effects of N-89 were confirmed in vitro; schistosomula of S. mansoni were killed by N-89 at an EC50 of 16 nM. In contrast, no significant reduction in worm burden was observed when N-89 was administered at 5 weeks postinfection in vivo. However, egg production was markedly suppressed by N-89 treatment at that time point. On microscopic observation, the intestine of N-89-treated female worms seemed to be empty compared with the control group, and the mean body length was significantly shorter than that of controls. Nutritional impairment in the parasite due to N-89 treatment was possible, and therefore quantification of hemozoin was compared between parasites with or without N-89 treatment. We found that the hemozoin content was significantly reduced in N-89 treated parasites compared with controls (P<0.001). The surface of adult worms was observed by scanning and transmission electron microscopy, but there were no apparent changes. Taken together, these observations suggested that N-89 has strong antischistosomal effects, probably through a unique mode of drug efficacy. As N-89 is less toxic to mammalian host animals, it is a possible drug candidate against schistosomiasis.
Subject(s)
Heterocyclic Compounds, 2-Ring/pharmacology , Schistosoma mansoni/drug effects , Schistosomiasis mansoni/drug therapy , Schistosomicides/pharmacology , Spiro Compounds/pharmacology , Administration, Oral , Animals , Female , Hemeproteins/analysis , Hemeproteins/metabolism , Heterocyclic Compounds, 2-Ring/therapeutic use , Humans , Male , Mice , Mice, Inbred BALB C , Parasite Egg Count , Parasitic Sensitivity Tests , Schistosoma mansoni/physiology , Schistosoma mansoni/ultrastructure , Schistosomiasis mansoni/parasitology , Schistosomicides/therapeutic use , Spiro Compounds/therapeutic use , Time Factors , Treatment OutcomeABSTRACT
N-89, a new antimalarial endoperoxide, was selected as a promising antimalarial compound showing high activity and selectivity. To study the mechanism of N-89 action, N-89 resistant strain (NRC10) was obtained by intermittent drug pressure. NRC10 had a tenfold increase in the EC(50) value of N-89. No cross-resistance was obtained with other antimalarial compounds. Comparative proteome analysis of N-89 sensitive and NRC10 strains revealed over-expression of 12 spots and down-regulation of 14 spots in NRC10. Fifteen proteins were identified of Plasmodium falciparum origin. The identified proteins representing several functions, mainly related to the glycolytic pathway, and metabolism of protein and lipid. Our results suggest that identified proteins may be candidates of antimalarial endoperoxide targets.
Subject(s)
Antimalarials/pharmacology , Peroxides/pharmacology , Plasmodium falciparum/chemistry , Plasmodium falciparum/drug effects , Proteome/drug effects , Protozoan Proteins/analysis , Animals , Drug Resistance , Electrophoresis, Gel, Two-Dimensional , Parasitic Sensitivity TestsABSTRACT
In the competitive peroxidation of mixtures of two alkenes with Co(II)/O(2)/Et(3)SiH, it was found that the relative reactivities of the alkene substrates are influenced by three major factors:. (1) relative stability of the intermediate carbon-centered radical formed by the reaction of the alkene with HCo(III) complex, (2) steric effects around the C=C double bond, and (3) electronic factors associated with the C=C double bond. Consistent with results from simple alkenes, the chemo- and regioselective peroxidation of dienes was also realized. Depending on the diene structure, the product included not only the expected acyclic unsaturated triethylsilyl peroxides but also 1,2-dioxolane and 1,2-dioxane derivatives via intramolecular cyclization of the unsaturated peroxy radical intermediates.
Subject(s)
Alkadienes/chemistry , Antimalarials/chemical synthesis , Artemisinins/chemistry , Peroxides/chemical synthesis , Sesquiterpenes/chemistry , Crystallography, X-Ray , Cyclization , Molecular Conformation , Molecular StructureABSTRACT
Generation of singlet and triplet 2-silylcyclopentane-1,3-diyls and their reactivity have been investigated in the thermal and photochemical denitrogenation of 2,3-diaza-7-silylbicyclo[2.2.1]hept-2-ene. 5-Silylcyclopentene (silyl migration product) is quantitatively obtained, while 5-silylbicyclo[2.1.0]pentane (intramolecular ring-closure product) is not detected in the denitrogenation reactions. Deuterium labeling studies clarify that 5-silylcyclopentene is formed by a suprafacial [1,2] silyl migration in singlet 2-silylcyclopentane-1,3-diyl. UDFT calculations closely reproduce the observed reactivity of the singlet diradical: The enthalpic barriers of the intramolecular ring-closure are calculated to be DeltaH++exo468 = 5.8 kcal/mol and DeltaH++endo468 = 6.7 kcal/mol, which are much higher than the energy barrier for the [1,2] silyl migration, DeltaH++468 = 2.7 kcal/mol. The notable effect of the silyl group on raising the energy barrier of the intramolecular cyclization is rationalized by an electronic configuration of the lowest singlet state of 2-silylcyclopentane-1,3-diyls.
ABSTRACT
Temperature and substituent effects on the stereo- and regioselectivity have been investigated in the photochemical [2 + 2] cycloaddition reaction, the so-called Paternò-Büchi (PB) reaction, of unsymmetrically substituted furans 2a,b (2-methyl- and 3-methylfuran) with aromatic carbonyl compounds 1a,b (benzaldehyde and benzophenone). The regio-random but stereoselective (exo/endo > 97/3) formation of lower substituted oxetane 3a and higher substituted oxetane 4a is found in the reaction with benzaldehyde (1a). The exclusive stereoselectivity is not dependent on the position of methyl substituent on the furan ring and the reaction temperature. The double-bond selection (3a versus 4a) is slightly dependent on the reaction temperature (3a/4a = 55/45 to 40/60). The Eyring plots of the regioselectivity are linear. Contrastively, in the reaction with benzophenone (1b), the double-bond selection (3b versus 4b) largely depends on the reaction temperature. The Eyring plots are not linear, but the inflection points are observed. The transient absorption spectroscopic analyses (picosecond time scale) clarify the intervention of triplet 2-oxabutane-1,4-diyls in the photochemical processes. Computational studies reveal the equilibrium structures of the triplet diradicals, energy barriers between the conformers, and the equilibrium constants. A rational mechanism is herein proposed by the support of both experimental and computational investigations to account for not only the exclusive formation of the exo-configured oxetanes 3a and 4a but also the nonlinear Eyring plots observed in the reaction with 1b.
ABSTRACT
The effect of a 2,2-ethylene-ketal functionality on the singlet-triplet energy gap (Delta E(ST)) and on the first electronic transition in singlet cyclopentane-1,3-diyls (1) has been investigated. UDFT calculations predict a significant increase in the preference for a singlet ground state in the diradical with the cyclic ketal at C2 (1g; Delta E(ST) = -6.6 kcal/mol in C(2) symmetry and -7.6 kcal/mol in C(2v) symmetry), compared to the 2,2-dihydroxy- and 2,2-dimethoxy-disubstituted diradicals (1d, Delta E(ST) = -3.6 kcal/mol in C(2) symmetry, and 1e, Delta E(ST) = -3.4 kcal/mol in C(2) symmetry). Spiroconjugation is shown to be responsible for the larger calculated value of absolute value Delta E(ST) in 1g, relative to 1d and 1e. A strong correlation between the calculated values of Delta E(ST) and the computed electronic excitation energies of the singlet diradicals is found for diradicals 1d, 1e, and 1g and for 2,2-difluorocyclopentane-1,3-diyl (1c). A similar correlation between Delta E(ST) and lambda(calcd) is predicted for the corresponding 1,3-diphenylcyclopentane-1,3-diyls 3, and the predicted blue shift in the spectrum of 3g, relative to 3e, has been confirmed by experimental comparisons of the electronic absorption spectra of the annelated derivatives 2c, 2e, and 2g in a glass at 77 K. The wavelength of the first absorption band in the singlet diradicals decreases in the order 2e (lambda(onset) = 650 nm) > 2g (lambda(onset) = 590 nm) > 2c (lambda(onset) = 580 nm). The combination of these computational and experimental results provides a sound basis for reassignment of the first electronic absorption band in singlet diradicals 2c, 2e, and 2g to the excitation of an electron from the HOMO to the LUMO of these 2,2-disubstituted derivatives of cyclopentane-1,3-diyl.
ABSTRACT
A series of novel amphiphilic hydroperoxides, alpha-alkoxyalkyl hydroperoxides (alpha-AHPs) and their related hydroperoxides, were designed and prepared with the intention of developing new oxidizing agents bearing a micelle-forming character in water. After their fundamental physical and interfacial properties were elucidated, both oxidation of benzyl sulfide and epoxidation of geraniol promoted by these hydroperoxides were investigated in detail under various conditions. Effective oxidation ofbenzyl sulfide to the corresponding sulfoxide was achieved in aqueous micellar systems composed of alpha-AHPs la-d and a catalytic amount of MoO2(acac)2 at 30 degrees C. Up to 100% conversion was observed under the optimum conditions. In the case of epoxidation of geraniol in water, the corresponding 2,3-epoxide was selectively formed in good yields. Because the conversion of each substrate in the micellar systems was higher than that in nonmicellar media, the solubilization of substrates into micelles was certainly effective in promoting oxidations of insoluble substrates in aqueous media. Micellar oxidation systems composed of novel amphiphilic hydroperoxides afford a new protocol in order to derive safer conditions under which these reactions may be carried out.
Subject(s)
Peroxides/chemistry , Peroxides/chemical synthesis , Hydrogen-Ion Concentration , Micelles , Molecular Structure , Oxidation-Reduction , Surface Properties , Water/chemistryABSTRACT
Co(II)-catalyzed peroxidation of dienes including (S)-limonene in the presence of molecular oxygen and triethylsilane provided in each case the corresponding 2,3-dioxabicyclo[3.3.1]nonane derivatives via the intramolecular cyclization of the unsaturated peroxy radical intermediates. The product composition was remarkably influenced by the structure of the dienes, the nature of the solvents, and the concentration of the substrates and the catalyst. Some of the yingzhaosu A analogues obtained in this study showed notable antimalarial activities in vitro.
Subject(s)
Antimalarials/chemical synthesis , Peroxides/chemical synthesis , Sesquiterpenes/chemical synthesis , Alkadienes/chemistry , Animals , Cell Death/drug effects , Cell Line, Tumor , Cobalt , Mice , Oxidation-Reduction , Oxygen , Plasmodium falciparum/drug effects , Silanes , Structure-Activity RelationshipABSTRACT
Ozonolysis of (alkenyldioxy)cyclododecyl hydroperoxides in trifluoroethanol gave a separable mixture of the corresponding alpha-hydroperoxy- and alpha-hydroxy-substituted spiro-tetraoxacycloalkanes with ring sizes in the range 7-12. Dehydration of the hydroperoxides or oxidation of the hydroxy-compounds afforded the corresponding peroxylactones. The solid-state structure of 1,2,6,7-tetraoxaspiro[7.11]nonadecan-3-one was determined by X-ray crystallographic analysis.
ABSTRACT
Photooxygenation of 2-phenylnorbornene 1 in the presence of 30% aqueous hydrogen peroxide afforded 1,2-bishydroperoxide 3, which could be cycloalkylated on treatment with silver oxide and a 1,omega-diiodoalkane to provide the tricyclic peroxides 12. Trimethylsilylation of 3 followed by TMSOTf-catalyzed cyclocondensation with carbonyl compounds led to the formation of the tricyclic peroxides 14 containing a 1,2,4,5-tetroxepane structure. Photooxygenation of 1 in the presence of either unsaturated hydroperoxides or unsaturated alcohols followed by bis(collidine)iodine hexafluorophosphate promoted cyclization gave the corresponding cyclic peroxides 15-17. Several of these cyclic peroxides showed substantial antimalarial activity particularly in vitro.
Subject(s)
Antimalarials/chemical synthesis , Bridged Bicyclo Compounds, Heterocyclic/chemical synthesis , Peroxides/chemical synthesis , Animals , Antimalarials/chemistry , Antimalarials/pharmacology , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Bridged Bicyclo Compounds, Heterocyclic/pharmacology , Crystallography, X-Ray , Mice , Peroxides/chemistry , Peroxides/pharmacology , Plasmodium falciparum/drug effects , Tumor Cells, CulturedABSTRACT
A simple method for the determination of the absolute configuration of 1-arylethanthiols was achieved by the observation of an intramolecular CH/pi shielding effect in (1)H-NMR of the corresponding thiol esters. (S)-Isomer of the diastereomers always shows remarkable shielding effect.
Subject(s)
Nuclear Magnetic Resonance, Biomolecular/methods , Sulfhydryl Reagents/chemistry , Esters , Molecular Conformation , Protons , StereoisomerismABSTRACT
The spin-state-dependent reactivity, singlet versus triplet, of the 2-spiroepoxy-1,3-cyclopentane-1,3-diyl DR2 has been assessed through alcohol-trapping reactions for which the effect of solvent acidity on the product distribution of the alcohol trapping products 2 versus 3 + 4 and stereochemical deuterium-labeling studies have been performed. The proposed mechanism for the solvent effect on the product ratio (2/3 + 4) reveals the importance of the hydrogen-bonded intermediates I1 and I2 in the trapping reactions; the stereochemical deuterium-labeling results clarify the dipole structure trapped by the alcohol. The dipoles DP1 and DP2, in which the configuration between the epoxide oxygen and the deuterium atoms is retained, are inferred for the direct photodenitrogenation reactions (singlet state), whereas for the benzophenone-sensitized photoreactions (triplet state), after ISC, the ring-opened dipole DP3 is implied as the intermediate that is trapped by the alcohol.
ABSTRACT
DFT calculations (UB3LYP/6-31+G**) have been performed to predict the substituent effect on the ground-state spin-multiplicity and the singlet-triplet energy gap in cyclobutane-1,3-diyls, CB-DR. The ground state is calculated to be largely dependent on the substituents (X, Y) at the C2 and C4 positions. The substituent effects can be reasonably explained by the two sets of through-bond (TB) interactions which result from the coupling between the symmetric nonbonding molecular orbital (Psi(S)) and the C-X (Y) sigma and sigma* orbitals.
ABSTRACT
An improved synthetic route of OPC-29030, the platelet adhesion inhibitor, was established via the diastereoselective oxidation of a chiral non-racemic sulfide (R)-5 to (S(S))-6 by the catalytic oxidation using VO(acac)(2) and cumene hydroperoxide (CHP) in the presence of MS4A. Under the current condition, the diastereoselectivity was not influenced by the presence of moisture, and moderate to high selectivity (72% de) was obtained at -30 degrees C. The obtained sulfoxide, which diastereomeric excess was easily raised by the recrystallization, could successfully lead to OPC-29030.
Subject(s)
Platelet Adhesiveness/drug effects , Platelet Aggregation Inhibitors/chemical synthesis , Sulfides/chemical synthesis , Imidazoles , Oxidation-Reduction , Platelet Aggregation Inhibitors/chemistry , Quinolones , Stereoisomerism , Sulfides/chemistry , Sulfur CompoundsABSTRACT
A convenient method to determine the absolute configuration of trans-2-aryl cyclohexanols, 1-aryl alcohols and amines was achieved. This method takes advantage of the 1H NMR spectroscopic observations of the remarkable high-field shift of C18-CH3 protons caused by the aromatic shielding effect. It is based on a discrimination of the difference of the environments in two diastereomers derived from 3 beta-acetoxy-5-etienic acid. Furthermore, it was observed that the corresponding diastereomeric derivatives of the pyridyl alcohols were simply separated by extraction based on the difference in their basicity.
ABSTRACT
[reaction: see text] Both a Co(III)-alkyl complex and a Co(III)-alkylperoxo complex were found to catalyze triethylsilylperoxidation of alkenes with O(2) and Et(3)SiH. On this basis, together with the nonstereoselectivity in the Co(II)-catalyzed peroxidation of 3-phenylindene and the formation of the corresponding 1,2-dioxolane from 2-phenyl-1-vinylcyclopropane (a radical clock), we propose a reasonable mechanism for the Co(II)-catalyzed novel autoxidation of alkenes with Et(3)SiH discovered by Isayama and Mukaiyama.