RESUMEN
Cutaneous leishmaniasis (CL) is a vector-borne disease characterized by skin lesions that can evolve into high-magnitude ulcerated lesions. Thus, this study aimed to develop an innovative nanoemulsion (NE) with clove oil, Poloxamer® 407, and multiple drugs, such as amphotericin B (AmB) and paromomycin (PM), for use in the topical treatment of CL. METHODS: Droplet size, morphology, drug content, stability, in vitro release profile, in vitro cytotoxicity on RAW 264.7 macrophages, and antileishmanial activity using axenic amastigotes of Leishmania amazonensis were assessed for NEs. RESULTS: After optimizing the formulation parameters, such as the concentration of clove oil and drugs, using an experimental design, it was possible to obtain a NE with an average droplet size of 40 nm and a polydispersion index of 0.3, and these parameters were maintained throughout the 365 days. Furthermore, the NE showed stability of AmB and PM content for 180 days under refrigeration (4 °C), presented a pH compatible with the skin, and released modified AmB and PM. NE showed the same toxicity as free AmB and higher toxicity than free PM against RAW 264.7 macrophages. The same activity as free AmB, and higher activity than free PM against amastigotes L. amazonensis. CONCLUSION: It is possible to develop a NE for the treatment of CL; however, complementary studies regarding the antileishmanial activity of NE should be carried out.
Asunto(s)
Anfotericina B , Antiprotozoarios , Emulsiones , Leishmaniasis Cutánea , Paromomicina , Paromomicina/farmacología , Paromomicina/administración & dosificación , Anfotericina B/farmacología , Anfotericina B/administración & dosificación , Leishmaniasis Cutánea/tratamiento farmacológico , Leishmaniasis Cutánea/parasitología , Animales , Ratones , Antiprotozoarios/farmacología , Antiprotozoarios/administración & dosificación , Antiprotozoarios/química , Células RAW 264.7 , Macrófagos/efectos de los fármacos , Macrófagos/parasitología , Leishmania mexicana/efectos de los fármacos , Aceite de Clavo/farmacología , Aceite de Clavo/química , Poloxámero/química , Estabilidad de Medicamentos , Nanopartículas/químicaRESUMEN
Conventional treatments for cutaneous leishmaniasis, a neglected vector-borne infectious disease, can frequently lead to serious adverse effects. Paromomycin (PAR), an aminoglycoside antibiotic, has been suggested for the topical treatment of disease-related lesions, but even when formulated in high drug-loading dosage forms, presents controversial efficacy. The presence of five ionizable amino groups hinder its passive cutaneous penetration but make PAR an excellent candidate for iontophoretic delivery. The objective of this study was to verify the feasibility of using iontophoresis for cutaneous PAR delivery and to propose a topical passive drug delivery system that could be applied between iontophoretic treatments. For this, in vitro iontophoretic experiments evaluated different application durations (10, 30, and 360 min), current densities (0.1, 0.25, and 0.5 mA/cm2), PAR concentrations (0.5 and 1.0 %), and skin models (intact and impaired porcine skin). In addition, 1 % PAR hydrogel had its penetration profile compared to 15 % PAR ointment in passive transport. Results showed iontophoresis could deliver suitable PAR amounts to dermal layers, even in short times and with impaired skin. Biodistribution assays showed both iontophoretic transport and the proposed hydrogel delivered higher PAR amounts to deeper skin layers than conventional ointment, even though applying 15 times less drug. To our knowledge, this is the first report of PAR drug delivery enhancement by iontophoresis. In summary, the association of iontophoresis with a topical application of PAR gel seems appropriate for improving cutaneous leishmaniasis treatment.
Asunto(s)
Leishmaniasis Cutánea , Paromomicina , Animales , Porcinos , Paromomicina/metabolismo , Paromomicina/farmacología , Iontoforesis/métodos , Distribución Tisular , Pomadas/metabolismo , Piel/metabolismo , Administración Cutánea , Sistemas de Liberación de Medicamentos/métodos , Leishmaniasis Cutánea/tratamiento farmacológico , Hidrogeles/farmacologíaRESUMEN
The parasitic protozoa Leishmania (Leishmania) infantum is the etiological agent of human visceral leishmaniasis and canine leishmaniasis in South America, where Brazil is the most affected country. This zoonotic disease is transmitted by the bite of an infected phlebotomine sand fly and dogs constitute the main domestic reservoir of the parasite. In this study, we screened 2348 dogs of the municipality of Embu das Artes, Brazil, for antibodies against the parasite. Prevalence for canine leishmaniasis seropositivity was 2.81%, as assessed using a Dual-Path Platform rapid test for canine leishmaniasis. Twenty-five seropositive dogs were euthanized for parasite isolation and 14 isolates were successful obtained. Nucleotide sequencing of the internal transcribed spacer confirmed the isolates to be L. (L.) infantum, and very low sequence variability was observed among them. The in vitro susceptibility to miltefosine and paromomycin was assessed and moderate variation in paromomycin susceptibility was found among the isolates in the promastigote and intracellular amastigote stages. On the other hand, in vitro susceptibility to miltefosine of these isolates was homogenous, particularly in the amastigote stage (EC50 values from 0.69 to 2.07 µM). In addition, the miltefosine sensitivity locus was deleted in all the isolates, which does not corroborate the hypothesis that the absence of this locus is correlated with a low in vitro susceptibility. Our findings confirm that the municipality of Embu das Artes is endemic for canine leishmaniasis and that isolates from this region are susceptible to paromomycin and miltefosine, indicating the potential of these drugs to be clinically evaluated in the treatment of human visceral leishmaniasis in Brazil.
Asunto(s)
Enfermedades de los Perros , Leishmania infantum , Leishmaniasis Visceral , Animales , Brasil/epidemiología , Enfermedades de los Perros/parasitología , Perros , Humanos , Leishmaniasis Visceral/tratamiento farmacológico , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/veterinaria , Paromomicina/uso terapéuticoRESUMEN
Treatment of tegumentary leishmaniasis in Brazil is limited to pentavalent antimonial, amphotericin B and pentamidine. These drugs, administered parenterally, cause several side effects and have a varied clinical response, depending on the species of Leishmania. Urgent expansion of the therapeutic arsenal against the disease is therefore necessary. Paromomycin is an aminoglycoside antibiotic that has already been approved for the treatment of visceral leishmaniasis in Southeast Asia. Here, we provide an in vitro evaluation of the activity of paromomycin in fifteen clinical isolates from patients with tegumentary leishmaniasis at a reference center for the treatment of the disease. Furthermore, the in vitro susceptibility to this drug in reference strains of Leishmania species that are endemic in Brazil has also been evaluated. Among the clinical isolates, nine were typed as Leishmania (Viannia) braziliensis, five as L. (Leishmania) amazonensis and one as L. (V.) guyanensis. Although never exposed to paromomycin, we found variable susceptibility among these isolates and reference strains in promastigotes and intracellular amastigotes, with the drug being more active in the amastigote form of the parasite. This study provides a preclinical dataset that is useful for the evaluation of paromomycin in the treatment of tegumentary leishmaniasis caused by species that are endemic in Brazil.
Asunto(s)
Antiprotozoarios/farmacología , Leishmania/efectos de los fármacos , Leishmaniasis Cutánea/parasitología , Paromomicina/farmacología , Animales , Humanos , Leishmaniasis Cutánea/tratamiento farmacológico , Ratones , Ratones Endogámicos BALB CRESUMEN
Paromomycin is an aminoglycoside antibiotic approved in 2006 for the treatment of visceral leishmaniasis caused by Leishmania donovani in Southeast Asia. Although this drug is not approved for the treatment of visceral and cutaneous leishmaniasis in Brazil, it is urgent and necessary to evaluate the potential of this drug as alternative for the treatment against species responsible for these clinical forms of the disease. In Brazil, Leishmania amazonensis is responsible for cutaneous and diffuse cutaneous leishmaniasis. The diffuse cutaneous form of the disease is difficult to treat and frequent relapses are reported, mainly when the treatment is interrupted. Here, we evaluated paromomycin susceptibility in vitro of a L. amazonensis clinical isolate from a patient with cutaneous leishmaniasis and the reference strain L. amazonensis M2269, as well as its in vivo efficacy in a murine experimental model. Although never exposed to paromomycin, a significant differential susceptibility between these two lines was found. Paromomycin was highly active in vitro against the clinical isolate in both forms of the parasite, while its activity against the reference strain was less active. In vivo studies in mice infected with each one of these lines demonstrated that paromomycin reduces lesion size and parasite burden and a direct correlation between the susceptibility in vitro and the effectiveness of this drug in vivo was found. Our findings indicate that paromomycin efficacy in vivo is dependent on intrinsic susceptibility of the parasite. Beyond that, this study contributes for the evaluation of the potential use of paromomycin in chemotherapy of cutaneous leishmaniasis in Brazil caused by L. amazonensis.
Asunto(s)
Antiprotozoarios , Leishmania mexicana , Paromomicina/farmacología , Animales , Antiprotozoarios/farmacología , Antiprotozoarios/uso terapéutico , Brasil , Humanos , Leishmania mexicana/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Resultado del TratamientoRESUMEN
BACKGROUND: Cutaneous leishmaniasis (CL) presents as 1 or more skin lesions, which makes local therapy inherently attractive compared to systemic therapy that exposes the whole body to a drug. For 30 years, 15% paromomycin topical formulations have been in clinical experimentation. Recently, 15% paromomycin in Aquaphilic, a complex base to facilitate adsorption into the lesion, was found superior to aquaphilic vehicle for Old World Leishmania major disease. METHODS: We performed a randomized trial of 15% paromomycin in Aquaphilic (40 patients) vs Aquaphilic vehicle (20 patients) vs a positive control (intralesional pentamidine; 20 patients) against L. braziliensis CL in Bolivia. RESULTS: Cure rates after 6 months of follow-up were 31 of 40 (77.5%, 95% confidence interval [CI] 62.5-88%) for paromomycin-Aquaphilic, 2 of 20 (10%, 95% CI 3-30%) for Aquaphilic vehicle (P < .0001 vs paromomycin-Aquaphilic), and 14 of 20 (70%, 95% CI 48-85.5%) for intralesional pentamidine. Both paromomycin-Aquaphilic and the Aquaphilic vehicle were very well tolerated, with only grade 1 adverse reactions in 5-10% of patients. CONCLUSIONS: Against L. braziliensis CL, a prevalent, aggressive form of New World CL, 15% paromomycin-aquaphilic was vastly superior to a negative vehicle control and was comparable in efficacy to a positive control. This study enlarges the potential use of 15% paromomycin-Aquaphilic from one form of Old World CL to CL more generally. CLINICAL TRIALS REGISTRATION: NCT03096457.
Asunto(s)
Antiprotozoarios/uso terapéutico , Leishmania braziliensis , Leishmaniasis Cutánea/tratamiento farmacológico , Paromomicina/uso terapéutico , Administración Tópica , Adulto , Antiprotozoarios/administración & dosificación , Humanos , Paromomicina/administración & dosificación , Pentamidina/administración & dosificación , Pentamidina/uso terapéutico , Adulto JovenRESUMEN
Treatment of Visceral Leishmaniasis (VL), a neglected tropical disease, is very challenging with few treatment options. Long duration of treatment and drug toxicity further limit the target of achieving VL elimination. Chemotherapy remains the treatment of choice. Single dose of liposomal amphotericin B (LAmB) and multidrug therapy (LAmB + miltefosine, LAmB + paromomycin (PM), or miltefosine + PM) are recommended treatment regimen for treatment of VL in Indian sub-continent. Combination therapy of pentavalent antimonials (Sbv) and PM in East Africa and LAmB in the Mediterranean region/South America remains the treatment of choice. Various drugs having anti-leishmania properties are in preclinical phase and need further development. An effective treatment and secondary prophylaxis of HIV-VL co-infection should be developed to decrease treatment failure and drug resistance.
Asunto(s)
Antiprotozoarios/uso terapéutico , Leishmania donovani/efectos de los fármacos , Leishmaniasis Visceral/tratamiento farmacológico , Resultado del Tratamiento , Anfotericina B/farmacología , Anfotericina B/uso terapéutico , Anfotericina B/toxicidad , Animales , Antiprotozoarios/efectos adversos , Antiprotozoarios/farmacología , Antiprotozoarios/toxicidad , Ensayos Clínicos como Asunto , Resistencia a Múltiples Medicamentos , Quimioterapia Combinada , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/parasitología , Humanos , India/epidemiología , Leishmaniasis Visceral/epidemiología , Antimoniato de Meglumina/administración & dosificación , Antimoniato de Meglumina/efectos adversos , Antimoniato de Meglumina/uso terapéutico , Paromomicina/farmacología , Paromomicina/uso terapéutico , Paromomicina/toxicidad , Fosforilcolina/análogos & derivados , Fosforilcolina/farmacología , Fosforilcolina/uso terapéutico , Fosforilcolina/toxicidad , Psychodidae/parasitología , Ovinos , América del Sur/epidemiologíaRESUMEN
Background: Cryptosporidiosis is one of the most important problems among neonatal ruminants. Cryptosporidium parvum is the agent causing the disease. Cryptosporidium infection, responsible for diarrhea, dehydration, weight loss and death in neonatal ruminants, leads to significant economic losses for producers. In calves naturally or experimentally infected with cryptosporidiosis, many agents have been reported to have therapeutic and protective effects. The objective of this study was to compare the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum.Materials, Methods & Results: Twenty calves between 7 and 20 days of age and naturally infected were included in the study. There were two different treatment groups in the study. The first group of calves were administered 100 µg/kg/day halofuginone lactate for 7 days and the second group of calves were administered 100 mg/kg/day paromomycin for 7 days. In addition, antibiotic and vitamin C were applied to all calves. Jugular venous blood samples were obtained pre-treatment and on the 7th day after the treatment. Routine clinical examinations of the calves were performed on days 0, 1, 3, 5 and 7 and rectal stool samples were collected for the detection of Cryptosporidium oocysts. Severe diarrhea, dehydration, depression and weight loss were observed in calves in both treatment groups. A significant decrease (P < 0.05) was observed for both groups in oocyst count on days 3, 5 and 7 compared to days 0 and 1. Improvements in blood parameters, stool characters and clinical scoring of both groups were observed in post-treatment. Discussion: Many pharmaceuticals or compounds have been tested for animal with cryptosporidiosis and only very few of them have shown a partial protective activity in ruminants when used prophylactically. Halofuginone lactate and paromomycin are commonly recommended as both therapeutic and protective agents.[...]
Asunto(s)
Animales , Lactante , Bovinos , Criptosporidiosis/terapia , Cryptosporidium parvum , Lactatos/administración & dosificación , Lactatos/uso terapéutico , Paromomicina/uso terapéutico , Aminoglicósidos , QuinazolinonasRESUMEN
Background: Cryptosporidiosis is one of the most important problems among neonatal ruminants. Cryptosporidium parvum is the agent causing the disease. Cryptosporidium infection, responsible for diarrhea, dehydration, weight loss and death in neonatal ruminants, leads to significant economic losses for producers. In calves naturally or experimentally infected with cryptosporidiosis, many agents have been reported to have therapeutic and protective effects. The objective of this study was to compare the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum.Materials, Methods & Results: Twenty calves between 7 and 20 days of age and naturally infected were included in the study. There were two different treatment groups in the study. The first group of calves were administered 100 µg/kg/day halofuginone lactate for 7 days and the second group of calves were administered 100 mg/kg/day paromomycin for 7 days. In addition, antibiotic and vitamin C were applied to all calves. Jugular venous blood samples were obtained pre-treatment and on the 7th day after the treatment. Routine clinical examinations of the calves were performed on days 0, 1, 3, 5 and 7 and rectal stool samples were collected for the detection of Cryptosporidium oocysts. Severe diarrhea, dehydration, depression and weight loss were observed in calves in both treatment groups. A significant decrease (P < 0.05) was observed for both groups in oocyst count on days 3, 5 and 7 compared to days 0 and 1. Improvements in blood parameters, stool characters and clinical scoring of both groups were observed in post-treatment. Discussion: Many pharmaceuticals or compounds have been tested for animal with cryptosporidiosis and only very few of them have shown a partial protective activity in ruminants when used prophylactically. Halofuginone lactate and paromomycin are commonly recommended as both therapeutic and protective agents.[...](AU)
Asunto(s)
Animales , Lactante , Bovinos , Lactatos/administración & dosificación , Lactatos/uso terapéutico , Cryptosporidium parvum , Criptosporidiosis/terapia , Paromomicina/uso terapéutico , Aminoglicósidos , QuinazolinonasRESUMEN
Descrevem-se o efeito terapêutico e os eventos adversos associados com o uso tópico de paromomicina 10 por cento em gel na leishmaniose cutânea. Quinze pacientes com leishmaniose cutânea cumpriram os critérios de inclusão descritos a seguir: contra-indicação para o uso de antimoniato de meglumina, intradermorreação de Montenegro positiva e até quatro lesões ulceradas. A fórmula foi prescrita duas vezes ao dia por 20 dias. Quatorze pacientes estiveram disponíveis para a avaliação do desfecho terapêutico e a proporção de cura foi de 21,4 por cento (3/14), 50 por cento melhoraram até a epitelização completa e a proporção de falha foi de 28,6 por cento. Nove pacientes que não apresentaram cura inicialmente foram re-tratados. Oito receberam uma nova série de paromomicina tópica e um foi tratado com antimoniato de meglumina. Dois pacientes não receberam novo tratamento e tiveram melhora lenta e contínua. Cinco de oito pacientes retratados com paromomicina tópica alcançaram a cura clínica, e três apresentaram falha, incluindo um paciente que tinha apresentado melhora com o primeiro tratamento. Os eventos adversos foram leves e locais em 53,3 por cento dos pacientes e nunca levaram à suspensão do tratamento.
The therapeutic effect of and adverse events associated with topical use of 10 percent paromomycin gel on cutaneous leishmaniasis are described. Fifteen patients with cutaneous leishmaniasis fulfilled the following inclusion criteria: contraindication for the use of meglumine antimoniate, positive Montenegro skin test and up to four ulcerated lesions. The formula was prescribed twice a day for 20 days. Fourteen patients were available for the therapeutic outcome evaluation. The cure rate was 21.4 percent (3/14); 50 percent improved as far as complete epithelialization; and the failure rate was 28.6 percent. Nine patients who did not initially present cure were retreated. Eight received a new series of topical paromomycin and one was treated with meglumine antimoniate. Two patients did not receive any new treatment and had continuous slow improvement. Five out of the eight patients retreated with topical paromomycin achieved clinical cure, and three presented failure, including one patient who had shown any improvement with the first treatment. For 53.3 percent of the patients, the adverse events were mild and local and never led to treatment suspension.