RESUMEN
The fact that animal models fail to replicate human disease faithfully is now being widely accepted by researchers across the globe. As a result, they are exploring the use of alternatives to animal models. The time has come to refine our experimental practices, reduce the numbers and eventually replace the animals used in research with human-derived and human-relevant 3-D disease models. Oncoseek Bio-Acasta Health, which is an innovative biotechnology start-up company based in Hyderabad and Vishakhapatnam, India, organises an annual International Conference on 3Rs Research and Progress. In 2021, this conference was on 'Advances in Research Animal Models and Cutting-Edge Research in Alternatives'. This annual conference is a platform that brings together eminent scientists and researchers from various parts of the world, to share recent advances from their research in the field of alternatives to animals including new approach methodologies, and to promote practices to help refine animal experiments where alternatives are not available. This report presents the proceedings of the conference, which was held in hybrid mode (i.e. virtual and in-person) in November 2021.
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Experimentación Animal , Alternativas a las Pruebas en Animales , Alternativas a las Pruebas en Animales/métodos , Bienestar del Animal , Animales , Humanos , India , Modelos AnimalesRESUMEN
BACKGROUND: The US Army designed artelinate/lysine salt (AL) to overcome the instability of sodium artesunate in aqueous solution (AS). To select the most efficacious artemisinin treatment, direct comparison was performed in an uncomplicated non-human primate malaria model. METHODS: Splenectomized rhesus monkeys were inoculated with Plasmodium coatneyi and on day six, single equimolar loading dose of IV AL (11.8 mg kg(-1)) or IV AS (8 mg kg(-1)) were administered followed by 1/2 the first dose once daily for 2 more days. Blood smear were performed twice daily and the number of parasites were counted microscopically. Blood samples were obtained after the first dose within 6 h for pharmacokinetic (PK) and ex vivo pharmacodynamic evaluation by simultaneously measuring plasma drug concentration and anti-malarial activity against Plasmodium falciparum in vitro. RESULTS: The anti-P. coatneyi in vivo activity of both compounds were comparable, but the ex vivo anti-P. falciparum potency of the IV AS regimen as administered was sevenfold higher than that of IV AL. Comparing in vivo pharmacodynamics of AL and AS, daily assessed parasite counts showed comparable 99 % parasite clearance times (PC99: 2.03, 1.84 day), parasite clearance rates (5.34, 4.13 per min) and clearance half-life (PCt1/2: 7.79, 10.1 h). This study showed strong and significant inverse correlation between PCt1/2 and t1/2 of AS + DHA, and AUC0-∞ of DHA, and correlated with Vz of AS (r(2) > 0.7, p ≤ 0.002). Lastly, following IV AL, there was a modest inverse correlation between PCt1/2 and Cmax (r(2) 0.6, p ≤ 0.04). Although all tested monkeys recrudesced subsequently, two died following AL regimen before parasite clearance. While the aetiology of those deaths could not be definitively determined, pathologic evidence favoured a sepsis-like syndrome and suggested that severe malaria was more likely than drug toxicity. CONCLUSION: The model demonstrated that both AS and DHA contributed to the anti-malarial activity of IV AS, while IV AL activity was largely restricted to the parent drug. Parasite clearance was strongly and linearly dependent on drug exposure for both artemisinin regimens. However, IV AS had higher ex vivo potency against P. falciparum, leading to an IND filing for GMP manufactured AS in the United States.
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Antimaláricos/farmacología , Antimaláricos/farmacocinética , Artemisininas/farmacología , Artemisininas/farmacocinética , Malaria/tratamiento farmacológico , Animales , Antimaláricos/administración & dosificación , Artemisininas/administración & dosificación , Artesunato , Modelos Animales de Enfermedad , Macaca mulatta , Esplenectomía , Resultado del Tratamiento , Estados UnidosRESUMEN
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.
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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.
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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
BACKGROUND: Mirincamycin is a close analog of the drug clindamycin used to treat Plasmodium falciparum blood stages. The clinical need to treat Plasmodium vivax dormant liver stages and prevent relapse with a drug other than primaquine led to the evaluation of mirinicamycin against liver stages in animals. METHODS: cis-mirinicamycin and trans-mirinicamycin were evaluated as prophylaxis against early liver stages of Plasmodium berghei in mice and as antirelapse hypnozoiticides against Plasmodium cynomolgi in the Rhesus monkey (Macaca mulatta). RESULTS: Mirincamycin was very effective against early liver stages of P. berghei in mice: both cis and trans enantiomers were 90-100% causally prophylactic at 3.3 mg/kg/day for 3 days orally. Both cis and trans mirincamycin, however, failed to kill dormant liver stages (hypnozoites) in the P. cynomolgi infected Rhesus monkey, the only preclinical hypnozoite model. Mirincamycin enantiomers at 80 mg/kg/day for 7 days orally, a dose that generated exposures comparable to that seen clinically, did not prevent relapse in any of four monkeys. CONCLUSIONS: Although efficacy against early liver stages of P. berghei was thought to correlate with anti-hypnozoite activity in primates, for mirincamycin, at least, there was no correlation. The negative P. cynomolgi hypnozoite data from Rhesus monkeys indicates that mirincamycin is unlikely to have potential as a clinical anti-relapse agent.
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Antimaláricos/uso terapéutico , Clindamicina/análogos & derivados , Malaria/tratamiento farmacológico , Plasmodium cynomolgi , Animales , Profilaxis Antibiótica , Antimaláricos/administración & dosificación , Antimaláricos/farmacocinética , Clindamicina/administración & dosificación , Clindamicina/farmacocinética , Clindamicina/uso terapéutico , Modelos Animales de Enfermedad , Femenino , Macaca mulatta , Malaria/parasitología , Ratones , Ratones Endogámicos ICR , Parasitemia , Plasmodium vivax , RecurrenciaRESUMEN
OBJECTIVE: A series of novel, substituted tetracyclic benzothiazepines were designed and prepared in an effort to optimize the potency of this chemical class against drug-resistant strains of the malaria parasite. METHODS: Tetracyclic benzothiazepines bearing structural modification at seven distinct positions within the structure were synthesized in Knoevenagel condensation reactions followed by sequential intermolecular thio-Michael and then intramolecular imine formation reactions. Following purification and chemical characterization, the novel compounds were tested for in vitro efficacy against blood-stage P. falciparum and liver-stage P. berghei and also for in vivo efficacy against P. berghei. RESULTS: Benzothiazepines bearing structural modification at the sulfur atom and at the three carbocycles within the molecule were successfully synthesized. The majority of analogs inhibited bloodstage P. falciparum with submicromolar IC50 values. The potency of an 8-methoxy-substituted analog 12 exceeded that of chloroquine in all three P. falciparum strains tested. The parent benzothiazepine 1 possessed liver-stage activity, inhibiting P. berghei sporozoites infecting HepG2 cells with an IC50 of 106.4 nM and an IC90 of 408.9 nM, but failed to enhance the longevity of P. berghei infected mice compared to the controls. Compounds displayed modest toxicity toward HepG2 cells and were tolerated by mice at the highest dose tested, 640 mg/kg/dose once daily for three days. CONCLUSION: The tetracyclic benzothiazepine described, which inhibits P. berghei infected hepatic cells with an IC50 of 106.4 nM, would appear to warrant further investigation. Optimization of ADME properties may be required since the most active analogs are probably excessively lipophilic.
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Antimaláricos , Malaria , Animales , Ratones , Plasmodium falciparum , Antimaláricos/farmacología , Malaria/tratamiento farmacológico , Plasmodium berghei , HígadoRESUMEN
BACKGROUND: Tafenoquine is an 8-aminoquinoline being developed for radical cure (blood and liver stage elimination) of Plasmodium vivax. During monotherapy treatment, the compound exhibits slow parasite and fever clearance times, and toxicity in glucose-6-phosphate dehydrogenase (G6PD) deficiency is a concern. Combination with other antimalarials may mitigate these concerns. METHODS: In 2005, the radical curative efficacy of tafenoquine combinations was investigated in Plasmodium cynomolgi-infected naïve Indian-origin Rhesus monkeys. In the first cohort, groups of two monkeys were treated with a three-day regimen of tafenoquine at different doses alone and in combination with a three-day chloroquine regimen to determine the minimum curative dose (MCD). In the second cohort, the radical curative efficacy of a single-day regimen of tafenoquine-mefloquine was compared to that of two three-day regimens comprising tafenoquine at its MCD with chloroquine or artemether-lumefantrine in groups of six monkeys. In a final cohort, the efficacy of the MCD of tafenoquine against hypnozoites alone and in combination with chloroquine was investigated in groups of six monkeys after quinine pre-treatment to eliminate asexual parasites. Plasma tafenoquine, chloroquine and desethylchloroquine concentrations were determined by LC-MS in order to compare doses of the drugs to those used clinically in humans. RESULTS: The total MCD of tafenoquine required in combination regimens for radical cure was ten-fold lower (1.8 mg/kg versus 18 mg/kg) than for monotherapy. This regimen (1.8 mg/kg) was equally efficacious as monotherapy or in combination with chloroquine after quinine pre-treatment to eliminate asexual stages. The same dose of (1.8 mg/kg) was radically curative in combination with artemether-lumefantrine. Tafenoquine was also radically curative when combined with mefloquine. The MCD of tafenoquine monotherapy for radical cure (18 mg/kg) appears to be biologically equivalent to a 600-1200 mg dose in humans. At its MCD in combination with blood schizonticidal drugs (1.8 mg/kg), the maximum observed plasma concentrations were substantially lower than (20-84 versus 550-1,100 ng/ml) after administration of 1, 200 mg in clinical studies. CONCLUSIONS: Ten-fold lower clinical doses of tafenoquine than used in prior studies may be effective against P. vivax hypnozoites if the drug is deployed in combination with effective blood-schizonticidal drugs.
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Aminoquinolinas/administración & dosificación , Antimaláricos/administración & dosificación , Macaca mulatta/parasitología , Malaria/tratamiento farmacológico , Enfermedades de los Primates/tratamiento farmacológico , Aminoquinolinas/farmacocinética , Animales , Antimaláricos/farmacocinética , Combinación Arteméter y Lumefantrina , Artemisininas/administración & dosificación , Artemisininas/farmacocinética , Cloroquina/administración & dosificación , Cloroquina/farmacocinética , Cromatografía Liquida , Modelos Animales de Enfermedad , Combinación de Medicamentos , Quimioterapia Combinada/métodos , Etanolaminas/administración & dosificación , Etanolaminas/farmacocinética , Fluorenos/administración & dosificación , Fluorenos/farmacocinética , Humanos , Masculino , Espectrometría de Masas , Mefloquina/administración & dosificación , Mefloquina/farmacocinética , Plasma/química , Plasmodium cynomolgi , Quinina/administración & dosificación , Quinina/farmacocinética , Resultado del TratamientoRESUMEN
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.
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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.
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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
The antimalarial activity and pharmacology of a series of phenylthiazolyl-bearing hydroxamate-based histone deacetylase inhibitors (HDACIs) was evaluated. In in vitro growth inhibition assays approximately 50 analogs were evaluated against four drug resistant strains of Plasmodium falciparum. The range of 50% inhibitory concentrations (IC(50)s) was 0.0005 to >1 microM. Five analogs exhibited IC(50)s of <3 nM, and three of these exhibited selectivity indices of >600. The most potent compound, WR301801 (YC-2-88) was shown to cause hyperacetylation of P. falciparum histones, which is a marker for HDAC inhibition in eukaryotic cells. The compound also inhibited malarial and mammalian HDAC activity in functional assays at low nanomolar concentrations. WR301801 did not exhibit cures in P. berghei-infected mice at oral doses as high as 640 mg/kg/day for 3 days or in P. falciparum-infected Aotus lemurinus lemurinus monkeys at oral doses of 32 mg/kg/day for 3 days, despite high relative bioavailability. The failure of monotherapy in mice may be due to a short half-life, since the compound was rapidly hydrolyzed to an inactive acid metabolite by loss of its hydroxamate group in vitro (half-life of 11 min in mouse microsomes) and in vivo (half-life in mice of 3.5 h after a single oral dose of 50 mg/kg). However, WR301801 exhibited cures in P. berghei-infected mice when combined at doses of 52 mg/kg/day orally with subcurative doses of chloroquine. Next-generation HDACIs with greater metabolic stability than WR301801 may be useful as antimalarials if combined appropriately with conventional antimalarial drugs.
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Antimaláricos/farmacología , Inhibidores Enzimáticos/farmacología , Inhibidores de Histona Desacetilasas , Plasmodium/efectos de los fármacos , Animales , Aotidae , Resistencia a Medicamentos , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/farmacocinética , Ácidos Hidroxámicos/síntesis química , Ácidos Hidroxámicos/farmacocinética , Ácidos Hidroxámicos/farmacología , Técnicas In Vitro , Malaria/tratamiento farmacológico , Malaria Falciparum/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos ICR , Plasmodium berghei/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Relación Estructura-Actividad , Tiazoles/síntesis química , Tiazoles/farmacocinética , Tiazoles/farmacologíaRESUMEN
A series of acid-stable carboxamide derivatives of 2-guanidinoimidazolidinedione (5a-c and 6a-c) were prepared as potential malaria prophylactic and radical cure agents. The new compounds showed moderate to good causal prophylactic activity in mice infected with Plasmodium yoelii sporozoites. Three compounds were further tested for causal prophylactic activity in Rhesus monkeys infected with Plasmodium cynomolgi sporozoites, and all showed a delay in patency from 13 to 40 days at 30 mg/kg/day x 3 days by IM dosing. Two out of four compounds tested for radical curative activity in Rhesus showed cure at 30 mg/kg/day x 3 days. The other two compounds showed delay in relapse from 16 to 68 days. Conversion of new carboxamides (5 and 6) to s-triazine derivatives (7) was demonstrated in mouse and human microsomal preparations and in rat plasma. The results suggest the metabolites, s-triazine derivatives 7, may be the active species of the new carboxamides 5a-c and 6a-c prepared in this study.
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Antimaláricos/síntesis química , Guanidinas/síntesis química , Imidazolidinas/síntesis química , Malaria/tratamiento farmacológico , Malaria/prevención & control , Animales , Antimaláricos/química , Antimaláricos/farmacología , Guanidinas/química , Guanidinas/farmacología , Humanos , Imidazolidinas/química , Imidazolidinas/farmacología , Técnicas In Vitro , Ratones , Microsomas Hepáticos/metabolismo , Plasmodium cynomolgi/efectos de los fármacos , Plasmodium yoelii/efectos de los fármacos , Ratas , Relación Estructura-ActividadRESUMEN
AAALAC International is a private, nonprofit organization that promotes humane treatment of animals in science through a voluntary international accreditation program. AAALAC International accreditation is recognized around the world as a symbol of high quality animal care and use for research, teaching and testing, as well as promoting animal welfare. Animals owned by the institution that are used for research, teaching and testing are included as part of an accredited program. More than 990 animal care and use institutions in 42 countries around the world (more than 170 programs in 13 countries in the Pacific Rim region) have earned AAALAC International accreditation. The AAALAC International Council on Accreditation evaluates overall performance and all aspects of an animal care and use program, involving an in-depth, multilayered, confidential peer-review process. The evaluators (site visitors) consider compliance with applicable local animal legislation of the host country, institutional policies, and employ a customized approach for evaluating overall program performance using a series of primary standards that include the Guide for the Care and Use of Laboratory Animals, the Guide for the Care and Use of Agricultural Animals in Research and Teaching, or the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Purposes, Council of Europe (ETS 123), and supplemental Reference Resources, as applicable.
RESUMEN
In spite of the increasing trend for local authorities and international organization to follow the same ethical principles, the regulatory framework on the protection, care, and use of research animals is still heterogeneous. Differences across geopolitical areas exist not only at the legal level but, more important, also at the level of implementation of the existing regulations. In this diverse landscape, AAALAC International uses performance standards to assure harmonization of accredited animal care and use programs globally. Accredited programs must first comply with applicable legislation, which may establish different baseline levels, on top of which performance standards are to be applied for the different animal care and use program areas. To achieve this goal, AAALAC International has given its Council on Accreditation an international composition and perspective that allows better interpretation of the different scenarios. There are a number of challenges and opportunities that may impact the implementation of existing legislation and the accreditation of animal care and use programs across countries. Challenges may relate to financial constraints, administrative burden, or differences between compliance and best practices. Opportunites may be represented by other fields of animal research, the level of development of research in some countries, or increasing interinstitutional collaborations. A description of these challenges and opportunities in North America, Europe, and the Pacific Rim from the AAALAC International perspective is discussed in this article.
RESUMEN
Malaria vaccine RTS,S combined with thrombospondin-related anonymous protein (TRAP) and formulated with AS02A (RTS,S+TRAP/AS02A) is safe and immunogenic in adult humans and rhesus monkeys (Macaca mulatta). Here, RTS,S+TRAP/AS02A was administered on a 0-, 1-, and 3-month schedule to three cohorts of infant monkeys, along with adult comparators. Cohort 1 evaluated 1/5, 1/2, and full adult doses, with the first dose administration at one month of age; cohort 2 monkeys received full adult doses, with the first dose administration at one versus three months of age; and, cohort 3 compared infants gestated in mothers with or without previous RTS,S/AS02A immunization. Immunization site reactogenicity was mild. Some infants, including the phosphate-buffered saline only recipient, developed transient iron-deficiency anemia, which is considered a result of repeated phlebotomies. All RTS,S+TRAP/AS02A regimens induced vigorous antibody responses that persisted through 12 weeks after the last vaccine dose. Modest lymphoproliferative and ELISPOT (interferon-gamma and interleukin-5) responses, particularly to TRAP, approximated adult comparators. RTS,S+TRAP/AS02A was safe and well tolerated. Vigorous antibody production and modest, selective cell-mediated immune responses suggest that RTS,S+TRAP/AS02A may be immunogenic in human infants.
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Adyuvantes Inmunológicos/administración & dosificación , Antígenos de Superficie de la Hepatitis B/inmunología , Lípido A/análogos & derivados , Lípido A/administración & dosificación , Vacunas contra la Malaria/inmunología , Proteínas Protozoarias/inmunología , Saponinas/administración & dosificación , Vacunas Sintéticas/inmunología , Animales , Anticuerpos Antiprotozoarios/sangre , Femenino , Humanos , Inmunización , Interferón gamma/biosíntesis , Interleucina-5/biosíntesis , Activación de Linfocitos , Macaca mulatta , Vacunas contra la Malaria/administración & dosificación , Vacunas contra la Malaria/toxicidad , Vacunas Sintéticas/toxicidadRESUMEN
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.
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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
Shigellosis is a worldwide disease, characterized by abdominal pain, fever, vomiting, and the passage of blood- and mucus-streaked stools. Rhesus monkeys and other primates are the only animals that are naturally susceptible to shigellosis. A suitable animal model is required for the pre-clinical evaluation of vaccines candidates. In this study, the minimal dose of Shigella dysenteriae1 1617 strain required to produce dysentery in four of five (80% attack rate) monkeys using an escalating dose range for three groups [2 × 10(8) , 2 × 10(9) and 2 × 10(10) colony forming unit (CFU)] was determined. In addition, the monkeys were re-infected. The identified optimal challenge dose was 2 × 10(9) CFU; this dose elicited 60% protection in monkeys when they were re-challenged with a one log higher dose (2 × 10(10) CFU). The challenge dose, 2 × 10(10) CFU, produced severe dysentery in all monkeys, with one monkey dying within 24 h, elicited 100% protection when re-challenged with the same dose. All monkeys exhibited immune responses. This study concludes that the rhesus monkey model closely mimics the disease and immune response seen in humans and is a suitable animal model for the pre-clinical evaluation of Shigella vaccine candidates. Prior infection with the 1617 strain can protect monkeys against subsequent re-challenges with homologous strains.
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Disentería Bacilar/inmunología , Disentería Bacilar/prevención & control , Vacunas contra la Shigella/administración & dosificación , Shigella dysenteriae/inmunología , Animales , Anticuerpos Antibacterianos/biosíntesis , Anticuerpos Antibacterianos/sangre , Células Productoras de Anticuerpos/inmunología , Carga Bacteriana , Colon/patología , Citocinas/biosíntesis , Citocinas/sangre , Modelos Animales de Enfermedad , Relación Dosis-Respuesta Inmunológica , Disentería Bacilar/microbiología , Heces/microbiología , Femenino , Humanos , Inmunoglobulina A Secretora/biosíntesis , Macaca mulatta , Masculino , Shigella dysenteriae/clasificación , Shigella dysenteriae/patogenicidad , EstómagoRESUMEN
There remains a need for new drugs to prevent relapse of Plasmodium vivax or P. ovale infection. The relapsing primate malaria P. cynomolgi has been used for decades to assess drugs for anti-hypnozoite activity. After sporozoite inoculation and blood-stage cure of initial parasitemia with chloroquine, rhesus macaques were treated on subsequent relapses with chloroquine in conjunction with test regimens of approved drugs. Tested drugs were selected for known liver or blood-stage activity and were tested alone or in conjunction with low-dose primaquine. Tinidazole and pyrazinamide prevented relapse when used in conjunction with chloroquine and low-dose primaquine. Triamterene and tinidazole administered without primaquine achieved radical cure in some animals. All other tested drugs or combinations failed to prevent relapse. The rhesus macaque-P. cynomolgi model remains a useful tool for screening drugs with anti-hypnozoite activity. Tinidazole and pyrazinamide require further investigation as agents to enable dose reduction of primaquine.
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Antimaláricos/uso terapéutico , Macaca mulatta/parasitología , Malaria Vivax/tratamiento farmacológico , Plasmodium cynomolgi/efectos de los fármacos , Plasmodium cynomolgi/patogenicidad , Animales , Cloroquina/uso terapéutico , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Malaria Vivax/prevención & control , Parasitemia/prevención & control , Plasmodium vivax/crecimiento & desarrollo , Plasmodium vivax/patogenicidad , Primaquina/uso terapéutico , Pirazinamida/uso terapéutico , Prevención Secundaria , Esporozoítos/efectos de los fármacos , Tinidazol/uso terapéutico , Triantereno/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 1,7-bis(alkylamino)diazachrysene-based small molecule was previously identified as an inhibitor of the botulinum neurotoxin serotype A light chain metalloprotease. Subsequently, a variety of derivatives of this chemotype were synthesized to develop structure-activity relationships, and all are inhibitors of the BoNT/A LC. Three-dimensional analyses indicated that half of the originally discovered 1,7-DAAC structure superimposed well with 4-amino-7-chloroquinoline-based antimalarial agents. This observation led to the discovery that several of the 1,7-DAAC derivatives are potent in vitro inhibitors of Plasmodium falciparum and, in general, are more efficacious against CQ-resistant strains than against CQ-susceptible strains. In addition, by inhibiting ß-hematin formation, the most efficacious 1,7-DAAC-based antimalarials employ a mechanism of action analogous to that of 4,7-ACQ-based antimalarials and are well tolerated by normal cells. One candidate was also effective when administered orally in a rodent-based malaria model. Finally, the 1,7-DAAC-based derivatives were examined for Ebola filovirus inhibition in an assay employing Vero76 cells, and three provided promising antiviral activities and acceptably low toxicities.
Asunto(s)
Antibacterianos/síntesis química , Antimaláricos/síntesis química , Antivirales/síntesis química , Toxinas Botulínicas Tipo A/antagonistas & inhibidores , Crisenos/síntesis química , Ebolavirus/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Quinolinas/síntesis química , Animales , Anopheles/parasitología , Antibacterianos/química , Antibacterianos/farmacología , Antimaláricos/química , Antimaláricos/farmacología , Antivirales/química , Antivirales/farmacología , Línea Celular , Chlorocebus aethiops , Crisenos/química , Crisenos/farmacología , Hemoproteínas/antagonistas & inhibidores , Malaria/tratamiento farmacológico , Ratones , Modelos Moleculares , Plasmodium berghei , Quinolinas/química , Quinolinas/farmacología , Ratas , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
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.