RESUMEN
Ivermectin (IVM) could be used for malaria control as treated individuals are lethal to blood-feeding Anopheles, resulting in reduced transmission. Tafenoquine (TQ) is used to clear the liver reservoir of Plasmodium vivax and as a prophylactic treatment in high-risk populations. It has been suggested to use ivermectin and tafenoquine in combination, but the safety of these drugs in combination has not been evaluated. Early derivatives of 8-aminoquinolones (8-AQ) were neurotoxic, and ivermectin is an inhibitor of the P-glycoprotein (P-gp) blood brain barrier (BBB) transporter. Thus, there is concern that co-administration of these drugs could be neurotoxic. This study aimed to evaluate the safety and pharmacokinetic interaction of tafenoquine, ivermectin, and chloroquine (CQ) in Rhesus macaques. No clinical, biochemistry, or hematological outcomes of concern were observed. The Cambridge Neuropsychological Test Automated Battery (CANTAB) was employed to assess potential neurological deficits following drug administration. Some impairment was observed with tafenoquine alone and in the same monkeys with subsequent co-administrations. Co-administration of chloroquine and tafenoquine resulted in increased plasma exposure to tafenoquine. Urine concentrations of the 5,6 orthoquinone TQ metabolite were increased with co-administration of tafenoquine and ivermectin. There was an increase in ivermectin plasma exposure when co-administered with chloroquine. No interaction of tafenoquine on ivermectin was observed in vitro. Chloroquine and trace levels of ivermectin, but not tafenoquine, were observed in the cerebrospinal fluid. The 3''-O-demethyl ivermectin metabolite was observed in macaque plasma but not in urine or cerebrospinal fluid. Overall, the combination of ivermectin, tafenoquine, and chloroquine did not have clinical, neurological, or pharmacological interactions of concern in macaques; therefore, this combination could be considered for evaluation in human trials.
RESUMEN
Previously, ivermectin (1 to 10 mg/kg of body weight) was shown to inhibit the liver-stage development of Plasmodium berghei in orally dosed mice. Here, ivermectin showed inhibition of the in vitro development of Plasmodium cynomolgi schizonts (50% inhibitory concentration [IC50], 10.42 µM) and hypnozoites (IC50, 29.24 µM) in primary macaque hepatocytes when administered as a high dose prophylactically but not when administered in radical cure mode. The safety, pharmacokinetics, and efficacy of oral ivermectin (0.3, 0.6, and 1.2 mg/kg) with and without chloroquine (10 mg/kg) administered for 7 consecutive days were evaluated for prophylaxis or radical cure of P. cynomolgi liver stages in rhesus macaques. No inhibition or delay to blood-stage P. cynomolgi parasitemia was observed at any ivermectin dose (0.3, 0.6, and 1.2 mg/kg). Ivermectin (0.6 and 1.2 mg/kg) and chloroquine (10 mg/kg) in combination were well-tolerated with no adverse events and no significant pharmacokinetic drug-drug interactions observed. Repeated daily ivermectin administration for 7 days did not inhibit ivermectin bioavailability. It was recently demonstrated that both ivermectin and chloroquine inhibit replication of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in vitro Further ivermectin and chloroquine trials in humans are warranted to evaluate their role in Plasmodium vivax control and as adjunctive therapies against COVID-19 infections.
Asunto(s)
Antimaláricos/farmacología , Cloroquina/farmacología , Ivermectina/farmacología , Hígado/efectos de los fármacos , Malaria/tratamiento farmacológico , Plasmodium cynomolgi/efectos de los fármacos , Animales , Antimaláricos/sangre , Antimaláricos/farmacocinética , Disponibilidad Biológica , Cloroquina/sangre , Cloroquina/farmacocinética , Esquema de Medicación , Combinación de Medicamentos , Sinergismo Farmacológico , Femenino , Hepatocitos/efectos de los fármacos , Hepatocitos/parasitología , Ivermectina/sangre , Ivermectina/farmacocinética , Hígado/parasitología , Macaca mulatta , Malaria/parasitología , Masculino , Parasitemia/tratamiento farmacológico , Plasmodium cynomolgi/crecimiento & desarrollo , Plasmodium cynomolgi/patogenicidad , Cultivo Primario de Células , Esquizontes/efectos de los fármacos , Esquizontes/crecimiento & desarrolloRESUMEN
Hematotoxicity in individuals genetically deficient in glucose-6-phosphate dehydrogenase (G6PD) activity is the major limitation of primaquine (PQ), the only antimalarial drug in clinical use for treatment of relapsing Plasmodium vivax malaria. PQ is currently clinically used in its racemic form. A scalable procedure was developed to resolve racemic PQ, thus providing pure enantiomers for the first time for detailed preclinical evaluation and potentially for clinical use. These enantiomers were compared for antiparasitic activity using several mouse models and also for general and hematological toxicities in mice and dogs. (+)-(S)-PQ showed better suppressive and causal prophylactic activity than (-)-(R)-PQ in mice infected with Plasmodium berghei. Similarly, (+)-(S)-PQ was a more potent suppressive agent than (-)-(R)-PQ in a mouse model of Pneumocystis carinii pneumonia. However, at higher doses, (+)-(S)-PQ also showed more systemic toxicity for mice. In beagle dogs, (+)-(S)-PQ caused more methemoglobinemia and was toxic at 5 mg/kg of body weight/day given orally for 3 days, while (-)-(R)-PQ was well tolerated. In a novel mouse model of hemolytic anemia associated with human G6PD deficiency, it was also demonstrated that (-)-(R)-PQ was less hemolytic than (+)-(S)-PQ for the G6PD-deficient human red cells engrafted in the NOD-SCID mice. All these data suggest that while (+)-(S)-PQ shows greater potency in terms of antiparasitic efficacy in rodents, it is also more hematotoxic than (-)-(R)-PQ in mice and dogs. Activity and toxicity differences of PQ enantiomers in different species can be attributed to their different pharmacokinetic and metabolic profiles. Taken together, these studies suggest that (-)-(R)-PQ may have a better safety margin than the racemate in human.
Asunto(s)
Antimaláricos/farmacocinética , Hemólisis/efectos de los fármacos , Malaria/tratamiento farmacológico , Neumonía por Pneumocystis/tratamiento farmacológico , Primaquina/farmacocinética , Animales , Antimaláricos/aislamiento & purificación , Antimaláricos/toxicidad , Perros , Transfusión de Eritrocitos , Eritrocitos/efectos de los fármacos , Eritrocitos/parasitología , Femenino , Deficiencia de Glucosafosfato Deshidrogenasa/metabolismo , Humanos , Dosificación Letal Mediana , Malaria/parasitología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Ratones Endogámicos NOD , Ratones SCID , Plasmodium berghei/efectos de los fármacos , Plasmodium berghei/fisiología , Pneumocystis carinii/efectos de los fármacos , Pneumocystis carinii/fisiología , Neumonía por Pneumocystis/microbiología , Primaquina/aislamiento & purificación , Primaquina/toxicidad , Estereoisomerismo , Trasplante HeterólogoRESUMEN
Renewed global efforts toward malaria eradication have highlighted the need for novel antimalarial agents with activity against multiple stages of the parasite life cycle. We have previously reported the discovery of a novel class of antimalarial compounds in the imidazolopiperazine series that have activity in the prevention and treatment of blood stage infection in a mouse model of malaria. Consistent with the previously reported activity profile of this series, the clinical candidate KAF156 shows blood schizonticidal activity with 50% inhibitory concentrations of 6 to 17.4 nM against P. falciparum drug-sensitive and drug-resistant strains, as well as potent therapeutic activity in a mouse models of malaria with 50, 90, and 99% effective doses of 0.6, 0.9, and 1.4 mg/kg, respectively. When administered prophylactically in a sporozoite challenge mouse model, KAF156 is completely protective as a single oral dose of 10 mg/kg. Finally, KAF156 displays potent Plasmodium transmission blocking activities both in vitro and in vivo. Collectively, our data suggest that KAF156, currently under evaluation in clinical trials, has the potential to treat, prevent, and block the transmission of malaria.
Asunto(s)
Antimaláricos/farmacología , Imidazoles/farmacología , Malaria Falciparum/tratamiento farmacológico , Malaria Falciparum/transmisión , Piperazinas/farmacología , Animales , Concentración 50 Inhibidora , Ratones , Ratones Endogámicos ICR , Plasmodium falciparum/efectos de los fármacos , Esporozoítos/efectos de los fármacosRESUMEN
WR319691 has been shown to exhibit reasonable Plasmodium falciparum potency in vitro and exhibits reduced permeability across MDCK cell monolayers, which as part of our screening cascade led to further in vivo analysis. Single-dose pharmacokinetics was evaluated after an IV dose of 5 mg/kg in mice. Maximum bound and unbound brain levels of WR319691 were 97 and 0.05 ng/g versus approximately 1,600 and 3.2 ng/g for mefloquine. The half-life of WR319691 in plasma was approximately 13 h versus 23 h for mefloquine. The pharmacokinetics of several N-dealkylated metabolites was also evaluated. Five of six of these metabolites were detected and maximum total and free brain levels were all lower after an IV dose of 5 mg/kg WR319691 compared to mefloquine at the same dose. These data provide proof of concept that it is feasible to substantially lower the brain levels of a 4-position modified quinoline methanol in vivo without substantially decreasing potency against P. falciparum in vitro.
Asunto(s)
Antimaláricos/metabolismo , Plasmodium falciparum/efectos de los fármacos , Quinolinas/metabolismo , Animales , Encéfalo/metabolismo , Hígado/metabolismo , Masculino , RatonesRESUMEN
Utilizing mefloquine as a scaffold, a next generation quinoline methanol (NGQM) library was constructed to identify early lead compounds that possess biological properties consistent with the target product profile for malaria chemoprophylaxis while reducing permeability across the blood-brain barrier. The library of 200 analogs resulted in compounds that inhibit the growth of drug sensitive and resistant strains of Plasmodium falciparum. Herein we report selected chemotypes and the emerging structure-activity relationship for this library of quinoline methanols.
Asunto(s)
Antimaláricos/síntesis química , Plasmodium falciparum/efectos de los fármacos , Quinolinas/síntesis química , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/farmacología , Animales , Antimaláricos/química , Antimaláricos/farmacología , Farmacorresistencia Fúngica , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Plasmodium falciparum/crecimiento & desarrollo , Quinolinas/química , Quinolinas/farmacología , Relación Estructura-ActividadRESUMEN
Hypnozoites are the liver stage non-dividing form of the malaria parasite that are responsible for relapse and acts as a natural reservoir for human malaria Plasmodium vivax and P. ovale as well as a phylogenetically related simian malaria P. cynomolgi. Our understanding of hypnozoite biology remains limited due to the technical challenge of requiring the use of primary hepatocytes and the lack of robust and predictive in vitro models. In this study, we developed a malaria liver stage model using 3D spheroid-cultured primary hepatocytes. The infection of primary hepatocytes in suspension led to increased infectivity of both P. cynomolgi and P. vivax infections. We demonstrated that this hepatic spheroid model was capable of maintaining long term viability, hepatocyte specific functions and cell polarity which enhanced permissiveness and thus, permitting for the complete development of both P. cynomolgi and P. vivax liver stage parasites in the infected spheroids. The model described here was able to capture the full liver stage cycle starting with sporozoites and ending in the release of hepatic merozoites capable of invading simian erythrocytes in vitro. Finally, we showed that this system can be used for compound screening to discriminate between causal prophylactic and cidal antimalarials activity in vitro for relapsing malaria.
Asunto(s)
Antimaláricos/farmacología , Hepatocitos/parasitología , Malaria/tratamiento farmacológico , Plasmodium/efectos de los fármacos , Animales , Técnicas de Cultivo de Célula/métodos , Línea Celular , Células Cultivadas , Hepatocitos/citología , Humanos , Hígado/citología , Hígado/parasitología , Macaca fascicularis , Macaca mulatta , Pruebas de Sensibilidad Parasitaria/métodos , Recurrencia , Prevención Secundaria , Esferoides Celulares/citología , Esferoides Celulares/parasitología , Esporozoítos/efectos de los fármacosRESUMEN
As part of the global effort toward malaria eradication, phenotypic whole-cell screening revealed the 2-aminopyridine class of small molecules as a good starting point to develop new antimalarial drugs. Stemming from this series, we found that the derivative, MMV390048, lacked cross-resistance with current drugs used to treat malaria. This compound was efficacious against all Plasmodium life cycle stages, apart from late hypnozoites in the liver. Efficacy was shown in the humanized Plasmodium falciparum mouse model, and modest reductions in mouse-to-mouse transmission were achieved in the Plasmodium berghei mouse model. Experiments in monkeys revealed the ability of MMV390048 to be used for full chemoprotection. Although MMV390048 was not able to eliminate liver hypnozoites, it delayed relapse in a Plasmodium cynomolgi monkey model. Both genomic and chemoproteomic studies identified a kinase of the Plasmodium parasite, phosphatidylinositol 4-kinase, as the molecular target of MMV390048. The ability of MMV390048 to block all life cycle stages of the malaria parasite suggests that this compound should be further developed and may contribute to malaria control and eradication as part of a single-dose combination treatment.
Asunto(s)
1-Fosfatidilinositol 4-Quinasa/antagonistas & inhibidores , Aminopiridinas/uso terapéutico , Antimaláricos/uso terapéutico , Sulfonas/uso terapéutico , Aminopiridinas/farmacología , Animales , Antimaláricos/farmacología , Femenino , Malaria/tratamiento farmacológico , Malaria/enzimología , Masculino , Ratones , Ratones SCID , Pruebas de Sensibilidad Parasitaria , Plasmodium/efectos de los fármacos , Plasmodium/patogenicidad , Sulfonas/farmacologíaRESUMEN
Since the 1940s, the large animal model to assess novel causal prophylactic antimalarial agents has been the Plasmodium cynomolgi sporozoite-infected Indian-origin rhesus monkey. In 2009 the model was reassessed with 3 clinical standards: primaquine (PQ), tafenoquine (TQ), and atovaquone-proguanil. Both control monkeys were parasitemic on day 8 post-sporozoite inoculation on day 0. Primaquine at 1.78 mg base/kg/day on days (-1) to 8 protected 1 monkey and delayed parasitemia patency of the other monkey to day 49. Tafenoquine at 6 mg base/kg/day on days (-1) to 1 protected both monkeys. However, atovaquone-proguanil at 10 mg atovaquone/kg/day on days (-1) to 8 did not protect either monkey and delayed patency only to days 18-19. Primaquine and TQ at the employed regimens are proposed as appropriate doses of positive control drugs for the model at present.
Asunto(s)
Aminoquinolinas/farmacología , Antimaláricos/farmacología , Atovacuona/farmacología , Malaria/prevención & control , Plasmodium cynomolgi/efectos de los fármacos , Primaquina/farmacología , Proguanil/farmacología , Aminoquinolinas/farmacocinética , Aminoquinolinas/uso terapéutico , Animales , Antimaláricos/farmacocinética , Antimaláricos/uso terapéutico , Atovacuona/farmacocinética , Atovacuona/uso terapéutico , Modelos Animales de Enfermedad , Combinación de Medicamentos , Macaca mulatta , Malaria/tratamiento farmacológico , Parasitemia/tratamiento farmacológico , Parasitemia/prevención & control , Primaquina/farmacocinética , Primaquina/uso terapéutico , Proguanil/farmacocinética , Proguanil/uso terapéuticoRESUMEN
Immune sera from volunteers vaccinated in a blinded Phase 3 clinical trial with JE-VAX(®) and a new Japanese encephalitis virus (JEV) vaccine (IC51 or IXIARO), were tested for the ability to protect mice against lethal JEV challenge. Sera from IXIARO vaccinated subjects were pooled into four batches based on neutralizing antibody measured by plaque reduction neutralization test (PRNT(50) titer): high (â¼200), medium (â¼40-50), low (â¼20) and negative (<10). Pooled sera from JE-VAX(®) vaccinated subjects (PRNT(50) titerâ¼55) and pooled JEV antibody negative pre-vaccination sera were used as controls. Groups of ten 6- to 7-week-old female ICR mice were injected intraperitoneally with 0.5 ml of each serum pool diluted 1:2 or 1:10, challenged approximately 18 h later with a lethal dose of either JEV strain SA14 (genotype III) or strain KE-093 (genotype I) and observed for 21 days. All mice in the non-immune serum groups developed clinical signs consistent with JEV infection or died, whereas high titer sera from both IXIARO and JE-VAX(®) sera protected 90-100% of the animals. Statistical tests showed similar protection against both JEV strains SA14 and KE-093 and protection correlated with the anti-JEV antibody titer of IXIARO sera as measured by PRNT(50). Ex vivo neutralizing antibody titers showed that almost all mice with a titer of 10 or greater were fully protected. In a separate study, analysis of geometric mean titers (GMTs) of the groups of mice vaccinated with different doses of IXIARO and challenged with JEV SA14 provided additional evidence that titers≥10 were protective.
Asunto(s)
Anticuerpos Neutralizantes/sangre , Anticuerpos Antivirales/sangre , Virus de la Encefalitis Japonesa (Especie)/inmunología , Encefalitis Japonesa/prevención & control , Vacunas contra la Encefalitis Japonesa/administración & dosificación , Animales , Virus de la Encefalitis Japonesa (Especie)/patogenicidad , Encefalitis Japonesa/inmunología , Encefalitis Japonesa/mortalidad , Encefalitis Japonesa/virología , Femenino , Humanos , Sueros Inmunes/administración & dosificación , Sueros Inmunes/inmunología , Inmunización , Inmunización Pasiva , Vacunas contra la Encefalitis Japonesa/inmunología , Masculino , Ratones , Ratones Endogámicos ICR , Pruebas de Neutralización , Análisis de Supervivencia , Resultado del TratamientoRESUMEN
A library of diamine quinoline methanols were designed based on the mefloquine scaffold. The systematic variation of the 4-position amino alcohol side chain led to analogues that maintained potency while reducing accumulation in the central nervous system (CNS). Although the mechanism of action remains elusive, these data indicate that the 4-position side chain is critical for activity and that potency (as measured by IC(90)) does not correlate with accumulation in the CNS. A new lead compound, (S)-1-(2,8-bis(trifluoromethyl)quinolin-4-yl)-2-(2-(cyclopropylamino)ethylamino)ethanol (WR621308), was identified with single dose efficacy and substantially lower permeability across MDCK cell monolayers than mefloquine. This compound could be appropriate for intermittent preventative treatment (IPTx) indications or other malaria treatments currently approved for mefloquine.
Asunto(s)
Antimaláricos/síntesis química , Etanolaminas/síntesis química , Malaria/prevención & control , Metanol/análogos & derivados , Metanol/síntesis química , Plasmodium falciparum/efectos de los fármacos , Quinolinas/síntesis química , Animales , Antimaláricos/farmacología , Línea Celular , Permeabilidad de la Membrana Celular , Dimerización , Perros , Resistencia a Medicamentos , Etanolaminas/farmacología , Etilenodiaminas/síntesis química , Etilenodiaminas/farmacología , Mefloquina/análogos & derivados , Mefloquina/síntesis química , Mefloquina/farmacología , Metanol/farmacología , Ratones , Plasmodium berghei , Quinolinas/farmacología , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
Febrifugine is the active principal isolated 50 years ago from the Chinese herb chang shan (Dichroa febrifuga Lour), which has been used as an antimalarial in Chinese traditional medicine for more than 2,000 years. However, intensive study of the properties of febrifugine has been hindered for decades due to its side effects. We report new findings on the effects of febrifugine analogs compared with those of febrifugine extracted from the dry roots of D. febrifuga. The properties of the extracted febrifugine were comparable to those obtained from the standard febrifugine provided by our collaborators. A febrifugine structure-based computer search of the Walter Reed Chemical Information System identified 10 analogs that inhibited parasite growth in vitro, with 50% inhibitory concentrations ranging from 0.141 to 290 ng/ml. The host macrophages (J744 cells) were 50 to 100 times less sensitive to the febrifugine analogs than the parasites. Neuronal (NG108) cells were even more insensitive to these drugs (selectivity indices, >1,000), indicating that a feasible therapeutic index for humans could be established. The analogs, particularly halofuginone, notably reduced parasitemias to undetectable levels and displayed curative effects in Plasmodium berghei-infected mice. Recrudescence of the parasites after treatment with the febrifugine analogs was the key factor that caused the death of most of the mice in groups receiving an effective dose. Subcutaneous treatments with the analogs did not cause irritation of the gastrointestinal tract when the animals were treated with doses within the antimalarial dose range. In summary, these analogs appear to be promising lead antimalarial compounds that require intensive study for optimization for further down-selection and development.