Renal Drug Transporters – Clinical Significance and Role in Drug-Drug Interactions

Thesis (Ph.D.)--University of Washington, 2016-07

Liver glucose-6-phosphatase proteins in suckling and weaned grey seal pups: structural similarities to other mammals and relationship to nutrition, insulin signalling and metabolite levels

Phocid seals have been proposed as models for diabetes because they exhibit limited insulin response to glucose, high blood glucose and increasing insulin resistance when fasting. Liver glucose-6-phosphatase (G6Pase) catalyses the final step in glucose production and is central to glucose regulation in other animals. G6Pase comprises a translocase (SLC37A4) and a catalytic subunit (G6PC). G6PC and SLC37A4 expression and activity are normally regulated by nutritional state and glucostatic hormones, particularly insulin, and are elevated in diabetes. We tested the hypotheses that (1) grey seal G6PC and SLC37A4 cDNA and predicted protein sequences differ from other species’ at functional sites, (2) relative G6Pase protein abundances are lower during feeding than fasting and (3) relative G6Pase protein abundances are related to insulin, insulin receptor phosphorylation and key metabolite levels. We show that G6PC and partial SLC37A4 cDNA sequences encode proteins sharing 82–95 % identity with other mammals. Seal G6PC contained no differences in sites responsible for activity, stability or subcellular location. Several substitutions in seal SLC37A4 were predicted to be tolerated with low probability, which could affect glucose production. Suckling pups had higher relative abundance of both subunits than healthy, postweaned fasting pups. Furthermore, relative G6PC abundance was negatively related to glucose levels. These findings contrast markedly with the response of relative hepatic G6Pase abundance to feeding, fasting, insulin, insulin sensitivity and key metabolites in other animals, and highlight the need to understand the regulation of enzymes involved in glucose control in phocids if these animals are to be informative models of diabetes.

Lifestyles of higher education students: the influence of self- esteem and psychological well-being

Background The transition to higher education can affect lifestyle-related factors. Objectives: To identify lifestyles of higher education students and analyse the influence of self-esteem and psychological well-being. Methods Correlational cross-sectional study. A total of 4,314 students partici- pated in the study. Online questionnaires were used: Estilo de Vida Fantástico (Fantastic Lifestyle Assessment) [1]; Questionário de Bem- estar Psicológico (Psychological General Well-Being Questionnaire) [2], and Escala de auto-estima de Rosenberg (Rosenberg Self-Esteem Scale [3]. Results Most students (85.3 %) have a healthy lifestyle. Lifestyle is strongly correlated with self-esteem and psychological well-being (p < 0.001). While analysing the association between self-esteem and psycho- logical well-being and the various lifestyle domains according to gen- der, a positive and significant correlation (p < 0.001) was found among female students, except for the Smoking domain (p = 0.393); in relation to psychological well-being, positive correlations were found in all domains. Among male students, positive and significant correlations (p < 0.001) were found in most lifestyle domains and self-esteem, except for the Smoking (p = 0.992), Alcohol and other drugs (p = 0.181) and Other behaviours (p = 0.442) domains; in rela- tion to psychological well-being, positive and significant correlations (p < 0.001) were found in most lifestyle domains, except for the Smoking (p = 0.458) and Other behaviours (p = 0.128) domains. Conclusions Based on the results, higher education institutions should support intervention projects to maintain high levels of psychological well- being and self-esteem, promoting healthy lifestyles.

Horse stable environment: what to expect regarding fungi and particles occupational exposure?

The simultaneous presence of fungi and particles in horse stable environment can create a singular exposure condition because particles have been reported has a good carrier for microorganisms and their metabolites. This study intends to characterize this setting and to recognize fungi and particles occupational exposure.

The metabolic and epigenetic effects of the isocitrate dehydrogenase-1 mutation in glioma

Cancer cells have been noted to have an altered metabolic phenotype for over ninety years. In the presence of oxygen, differentiated cells predominately utilise the tricarboxylic acid (TCA) cycle and oxidative phosphorylation to efficiently produce energy and the metabolites necessary for protein and lipid synthesis. However, in hypoxia, this process is altered and cells switch to a higher rate of glycolysis and lactate production to maintain their energy and metabolic needs. In cancer cells, glycolysis is maintained at a high rate, even in the presence of oxygen; a term described as “aerobic glycolysis”. Tumour cells are rapidly dividing and have a much greater need for anabolism compared to normal differentiated cells. Rapid glucose metabolism enables faster ATP production as well as a greater redistribution of carbons to nucleotide, protein, and fatty acid synthesis, thus maximising cell growth. Recently, other metabolic changes, driven by mutations in genes related to the TCA cycle, indicate an alternative role for metabolism in cancer, the “oncometabolite”. This is where a particular metabolite builds up within the cell and contributes to the tumorigenic process. One of these genes is isocitrate dehydrogenase (IDH) IDH is an enzyme that forms part of the tricarboxylic acid (TCA) cycle and converts isocitrate to α-ketoglutarate (α-KG). It exists in three isoforms; IDH1, IDH2 and IDH3 with the former present in the cytoplasm and the latter two in the mitochondria. Point mutations have been identified in the IDH1 and IDH2 genes in glioma which result in a gain of function by converting α-KG to 2-hydroxyglutarate (2HG), an oncometabolite. 2HG acts as a competitive inhibitor of the α-KG dependent dioxygenases, a superfamily of enzymes that are involved in numerous cellular processes such as DNA and histone demethylation. It was hypothesised that the IDH1 mutation would result in other metabolic changes in the cell other than 2HG production, and could potentially identify pathways which could be targeted for therapeutic treatment. In addition, 2HG can act as a potential competitive inhibitor of α-KG dependent dioxygenases, so it was hypothesised that there would be an effect on histone methylation. This may alter gene expression and provide a mechanism for tumourogenesis and potentially identify further therapeutic targets. Metabolic analysis of clinical tumour samples identified changes associated with the IDH1 mutation, which included a reduction in α-KG and an increase in GABA, in addition to the increase in 2HG. This was replicated in several cell models, where 13C labelled metabolomics was also used to identify a possible increase in metabolic flux from glutamate to GABA, as well as from α-KG to 2HG. This may provide a mechanism whereby the cell can bypass the IDH1 mutation as GABA can be metabolised to succinate in the mitochondria by GABA transaminase via the GABA shunt. JMJ histone demethylases are a subset of the α-KG dependent dioxygenases, and are involved in removing methyl groups from histone tails. Changes in histone methylation are associated with changes in gene expression depending on the site and extent of chemical modification. To identify whether the increase in 2HG and fall in α-KG was associated with inhibition of histone demethylases a histone methylation screen was used. The IDH1 mutation was associated with an increase in methylation of H3K4, which is associated with gene activation. ChiP and RNA sequencing identified an increase in H3K4me3 at the transcription start site of the GABRB3 subunit, resulting in an increase in gene expression. The GABRB3 subunit forms part of the GABA-A receptor, a chloride channel, which on activation can reduce cell proliferation. The IDH1 mutation was associated with an increase in GABA and GABRB3 subunit of the GABA-A receptor. This raises the possibility of GABA transaminase as a potential therapeutic target. Inhibition of this enzyme could reduce GABA metabolism, potentially reducing any beneficial effect of the GABA shunt in IDH1 mutant tumours, and increasing activation of the GABA-A receptor by increasing the concentration of GABA in the brain. This in turn may reduce cell proliferation, and could be achieved by using Vigabatrin, a GABA transaminase inhibitor licensed for use in epilepsy.

The effects of excise tax changes on alcoholic beverages and tobacco consumption in Thailand

The purpose of this thesis is to examine the factors affecting the likelihood of consumption and the amount spent on alcoholic beverages and tobacco in Thailand using the 2009 Socio-Economic Survey of Thailand. Results suggest that household size, tenure and occupation have significant impacts on both the probability of alcohol and tobacco consumption and spending levels. Income also plays a key role in explaining the amount spent on alcoholic beverages and tobacco. Demand elasticities are calculated under the Extended Linear Expenditure System (ELES). Demand for alcoholic beverages and tobacco is found to be inelastic. The effects of increasing taxation on alcohol and tobacco consumption in Thailand are estimated. The findings are that excise taxes in Thailand are efficient taxes with only a modest rise in deadweight loss. Taxes result in a small decrease in consumption but generate higher expenditure and government tax revenue. Excise tax on alcoholic beverages results in a net benefit to the Thai economy. Tobacco taxes increase total expenditure and government revenue as well as increasing net benefit to the Thai economy. However, the low elasticities of demand also mean that excise taxes have only a small impact on reducing the costs associated with drinking and smoking.

DEVELOPMENT AND REPRODUCIBILITY OF THE INSTRUMENTO DE AVALIAÇÃO DA PROMOÇÃO DA SAÚDE NA UNIVERSIDADE - IAPSU (ASSESSMENT TOOL FOR HEALTH PROMOTION AT THE UNIVERSITY)

Objective: To present the Instrumento de Avaliação da Promoção da Saúde na Universidade – IAPSU (Assessment Tool for Health Promotion at the University) and its reproducibility assessment process. Methods: Cross-sectional study conducted between May and July 2014 with 50 students from a university of Fortaleza, Ceará, which developed the IAPSU through the analysis of government documents and a systematic review of the literature on a potentially healthy university. The tool has 41 questions divided into five domains: physical activity, diet, environmental factors, psychosocial factors and alcohol and drug use, integrative and complementary practices. To assess the inter-examiner reproducibility, the students answered the IAPSU twice, applied by two different examiners; to assess the intraexaminer reproducibility, another application of the instrument was performed after seven days. Results: The study comprised 40 Nursing students and 10 Physical Therapy students, with a mean age of 25 ± 5.4 years; 88% were women and white individuals were predominant. In the reproducibility assessment, strong intraclass, intra- and inter-examiner correlation coefficients - above 0.8 - were observed in all the domains. Conclusion: The IAPSU is a reproducible and reliable instrument for assessing health promotion at the university.

ALCOHOL AS A SOCIAL MEDIATOR AMONG UNIVERSITY STUDENTS

Objective: To identify the prevalence of alcohol consumption in Psychology students of a higher education institution in the city of Montes Claros, MG. Methods: Quantitative crosssectional descriptive research conducted from September to October 2014. The population consisted of 116 Psychology students from the city of Montes Claros, MG. Data were collected using the Alcohol Use Disorders Identification Test (AUDIT), the Inventário de Expectativas e Crenças Pessoais Acerca do Álcool – IECPA (Inventory of Expectations and Personal Beliefs about Alcohol), the Alcohol, Smoking and Substance Involvement Screening Test (ASSIST) and the Escala de Satisfação com o Suporte Social – ESSS (Social Support Satisfaction Scale). Descriptive analysis of data was performed using SPSS 19.0. Results: The sample had a predominance of female gender (82.75%, n=96), pardos (65.51%, n=76) and single (60.34%, n=70) individuals. Regarding the AUDIT risk classification, it was found that 49.13% (n=57) of the respondents were in the level 4, considered alcohol dependence. They reported occasional use of alcohol, smoking and other substances, which refer to ASSIST level 1 classification, with 94.82% (n=110). Regarding the IECPA, 87.06% (n=101) of the individuals were classified as level 1, with low vulnerability to the effects of alcohol. As to the ESSS, 68.10% (n=79) of the students showed high social support. Conclusion: Regarding the sample studied, it was found a high prevalence of dependence on alcohol and other legal and illegal drugs.

In vitro studies of drug transformations: application to forensic toxicology

The forensic toxicologist faces challenges in the detection of drugs and poisons in biological samples due to transformations which occur both during life and after death. For example, changes can result from drug metabolism during life or from the use of formalin solution for post mortem embalming purposes. The former requires the identification of drug metabolites and the latter the identification of chemical reaction products in order to know which substances had been administered. The work described in this thesis was aimed at providing ways of tackling these challenges and was divided into two parts. Part 1 investigated the use of in vitro drug metabolism by human liver microsomes (HLM) to obtain information on drug metabolites and Part 2 investigated the chemical reactions of drugs and a carbamate pesticide with formalin solution and formalin-blood. The initial aim of part I was to develop an in vitro metabolism method using HLM, based on a literature review of previous studies of this type. MDMA was chosen as a model compound to develop the HLM method because its metabolism was known and standards of its metabolites were commercially available. In addition, a sensitive and selective method was developed for the identification and quantitation of hydrophilic phase I drug metabolites using LC/MS/MS with a conventional reverse-phase (C18) column. In order to obtain suitable retention factors for polar drug metabolites on this column, acetyl derivatives were evaluated for converting the metabolites to more lipophilic compounds and an optimal separation system was developed. Acetate derivatives were found to be stable in the HPLC mobile phase and to provide good chromatographic separation of the target analytes. In vitro metabolism of MDMA and, subsequently, of other drugs involved incubation of 4 µg drug substance in pH 7.4 buffer with an NADPH generating system (NGS) at 37oC for 90 min with addition of more NGS after 30 min. The reaction was stopped at 90 min by the addition of acetonitrile before extraction of the metabolites. Acetate derivatives of MDMA metabolites were identified by LC/MS/MS using multiple reaction monitoring (MRM). Three phase I metabolites (both major and minor metabolites) of MDMA were detected in HLM samples. 3,4-dihydroxy-methamphetamine and 4-hydroxy-3-methoxymethamphetamine were found to be major metabolites of MDMA whereas 3,4-methylenedioxyamphetamine was found to be a minor metabolite. Subsequently, ten MDMA positive urines were analysed to compare the metabolite patterns with those produced by HLM. An LC/MS method for MDMA and its metabolites in urine samples was developed and validated. The method demonstrated good linearity, accuracy and precision and insignificant matrix effects, with limits of quantitation of 0.025 µg/ml. Moreover, derivatives of MDMA and its metabolites were quantified in all 10 positive human urine samples. The urine metabolite pattern was found to be similar to that from HLM. The second aim of Part 1 was to use the HLM system to study the metabolism of some new psychoactive substances, whose misuse worldwide has necessitated the development of analytical methods for these drugs in biological specimens. Methylone and butylone were selected as representative cathinones and para-methoxyamphetamine (PMA) was chosen as a representative ring-substituted amphetamine, because of the involvement of these drugs in recent drug-related deaths, because of a relative lack of information on their metabolism, and because reference standards of their metabolites were not commercially available. An LC/MS/MS method for the analysis of methylone, butylone, PMA and their metabolites was developed. Three phase I metabolites of methylone and butylone were detected in HLM samples. Ketone reduction to β-OH metabolites and demethylenation to dihydroxy-metabolites were found to be major phase I metabolic pathways of butylone and methylone whereas N-demethylation to nor-methylone and nor-butylone were found to be minor pathways. Also, demethylation to para-hydroxyamphetamine was found to be a major phase I metabolic pathway of PMA whereas β-hydroxylation to β-OH-PMA was found to be a minor pathway. Formaldehyde is used for embalming, to reduce decomposition and preserve cadavers, especially in tropical countries such as Thailand. Drugs present in the body can be exposed to formaldehyde resulting in decreasing concentrations of the original compounds and production of new substances. The aim of part II of the study was to evaluate the in vitro reactions of formaldehyde with selected drug groups including amphetamines (amphetamine, methamphetamine and MDMA), benzodiazepines (alprazolam and diazepam), opiates (morphine, hydromorphone, codeine and hydrocodone) and with a carbamate insecticide (carbosulfan). The study would identify degradation products to serve as markers for the parent compounds when these were no longer detectable. Drugs standards were spiked in 10% formalin solution and 10% formalin blood. Water and whole blood without formalin were used for controls. Samples were analysed by LC/MS/MS at different times from the start, over periods of up to 30 days. Amphetamine, methamphetamine and MDMA were found to rapidly convert to methamphetamine, DMA and MDDMA respectively, in both formalin solution and formalin blood, confirming the Eschweiler-Clarke reaction between amine-containing compounds and formaldehyde. Alprazolam was found to be unstable whereas diazepam was found to be stable in both formalin solution and water. Both were found to hydrolyse in formalin solution and to give open-ring alprazolam and open-ring diazepam. Other alprazolam conversion products attached to paraformaldehyde were detected in both formalin solution and formalin blood. Morphine and codeine were found to be more stable than hydromorphone and hydrocodone in formalin solution. Conversion products of hydromorphone and hydrocodone attached to paraformaldehyde were tentatively identified in formalin solution. Moreover, hydrocodone and hydromorphone rapidly decreased within 24 h in formalin blood and could not be detected after 7 days. Carbosulfan was found to be unstable in formalin solution and was rapidly hydrolysed within 24 h, whereas in water it was stable up to 48 h. Carbofuran was the major degradation product, plus smaller amounts of other products, 3-ketocarbofuran and 3-hydrocarbofuran. By contrast, carbosulfan slowly hydrolysed in formalin-blood and was still detected after 15 days. It was concluded that HLM provide a useful tool for human drug metabolism studies when ethical considerations preclude their controlled administration to humans. The use of chemical derivatisation for hydrophilic compounds such as polar drug metabolites for analysis by LC/MS/MS with a conventional C18 column is effective and inexpensive, and suitable for routine use in the identification and quantitation of drugs and their metabolites. The detection of parent drugs and their metabolites or conversion and decomposition products is potentially very useful for the interpretation of cases in forensic toxicology, especially when the original compounds cannot be observed.

Substance Use and Academic Outcomes among High School Seniors: Examining the Influence of Emotional Academic Engagement

Substance use is prevalent among adolescents, with two-thirds trying alcohol and half trying an illicit drug by twelfth grade (Miech et al., 2015). Substance use is known to affect academic performance. This study utilized nationally representative data from the 2013 Monitoring the Future twelfth grade survey to examine the relationships between substance use, skipping school, grades, and academic engagement. One-quarter of respondents (26%) had never used a substance. The majority (67%) had used at least one substance during the past year. Substance use during their lifetime but not during the past year was uncommon (7%). Lifetime non-users were less likely than past-year users to skip school during the past month and to have low grades. Lifetime non-users also had greater academic self-efficacy and emotional academic engagement relative to past-year users. These findings underscore the importance of screening and intervention for substance use to promote academic achievement and adolescent wellbeing.

A survey on diversity and distribution of crabs in inter tidal and sub tidal shore of the Sistan and Baloochestan Province (Oman sea)

This project was done for identifying and survey on distribution and diversity of true crabs in inter tidal and sub tidal zones of the Gulf of Oman (Sistan and Baluchistan province) during two year from 2009 to 2010. Specimens of inter tidal zones were carried out at 10 stations and 8 stations from sub tidal. The specimens were collected by hand and dip-net from inter tidal and by trawl net from sub tidal regions, preserved in 70% alcohol and carried to the laboratory. A total of 37 species belonged to 17 families from inter tidal and 23 species belonged to 9 families from sub tidal were identified. Of which 54 species were identified up to species level. 2 species from Matutidae, 1 species from Eriphiidae, Menippidae, Pseudoziidae, Plagusidae, Varunidae, Camptandriidae, Dromiidae and Dorippidae, 2 species from Oziidae, 3 species from Epialtidae, 2 species from Majidae, 4 species from Pilumnidae, 12 species from Portunidae, 6 species from Xanthidae, 2 species from Grapsidae, 3 species from Dotillidae, 3 species from Macrophthalmidae, 3 species from Ocypodidae, 3 species from Calappidae, 2 species from Parthenopidae and 1 species from Galenidae were identified. All specimens are deposited in the Zoological Museum, University of Tehran (ZUTC). The results of the present study revealed that family Portunidae with 6 species from inter tidal and 9 species from sub tidal regions have the highest species richness among the 22 families. Maximum similarity (Sorenson's Index) was obtained among the stations Breis, Lipar, Pozm and Gordim, and minimum was obtained among the stations Chazire-Kharchang with Pasabandar, Beris, Lipar, Daria-Bozorg, Pozm and Gordim in intertidal regions. In sub tidal regions maximum similarity (Sorenson's Index) was obtained among the stations Pasa bandar with Berisand minimum was obtained among the stations Govatr with Ramin and Gordim, Ramin with Pozm. Also maximum species richness was observed at Tiss in inter tidal and Chabahar in sub tidal stations, whereas minimum was obtained at Beris, Pozm, Gordim and Lipar in inter tidal and Govatr and Pozm in sub tidal stations. Family Ocypodidae in inter tidal and Portunidae in sub tidal regions have the highest distribution. In all of the species length and Breadth of carapace showed significant relation.

Review: Head and neck squamous cell carcinoma in sub-Saharan Africa

Aim Review the literature from 1990 to 2013 to determine known anatomic sites, risk factors, treatments, and outcomes of head and neck squamous cell carcinoma (HNSCC) in sub-Saharan Africa. Methods Using a systematic search strategy, literature pertaining to HNSCC in sub-Saharan Africa was reviewed and patient demographics, anatomic sites, histology, stage, treatment, and outcomes were abstracted. The contributions of human immunodeficiency virus (HIV), human papillomavirus (HPV) and behavioural risk factors to HNSCC in the region were assessed. Results Of the 342 papers identified, 46 were utilized for review, including 8611 patients. In sub-Saharan Africa, the oropharyngeal/oral cavity was found to be the most common site, with 7750 cases (90% of all cases). Few papers distinguished oropharyngeal from oral cavity, making identification of possible HPV-associated oropharyngeal squamous cell carcinoma (SCC) difficult. SCC of the nasopharynx, nasal cavity, or paranasal sinuses was identified in 410 patients (4.8% of all cases). Laryngeal SCC was found in 385 patients (4.5% of all cases), and only 66 patients (0.8% of all cases) with hypopharyngeal SCC were identified. In 862 patients with data available, 43% used tobacco and 42% used alcohol, and reported use varied widely and was more common in laryngeal SCC than that of the oropharyngeal/oral cavity. Toombak and kola nut use was reported to be higher in patients with HNSCC. Several papers reported HIV-positive patients with HNSCC, but it was not possible to determine HNSCC prevalence in HIV-positive compared to negative patients. Reports of treatment and outcomes were rare. Conclusions The oropharyngeal/oral cavity was by far the most commonly reported site of HNSCC reported in sub-Saharan Africa. The roles of risk factors in HNSCC incidence in sub-Saharan Africa were difficult to delineate from the available studies, but a majority of patients did not use tobacco and alcohol.

Evaluation of the antioxidant activity and antitumor activity of marine invertebrates extracts

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologias, Universidade do Algarve, 2014

Toxicological impact of JWH-018 and its phase I metabolite N-(3-hydroxypentyl) on human cell lines

"The emergence and abuse of synthetic cannabinoids has been increasing as an alternative to cannabis, mainly among youth. As their appearance on the drug market has been recent, the pharmacological and toxicological profiles of these psychoactive substances are poorly understood. Current studies suggest that they have stronger effects compared to their natural alternatives and their metabolites retain affinity towards CB1 receptors in CNS. Since studies on its toxicological properties are scarce, the effects of the drug in human derived cell lines were investigated. The present study was designed to explore the toxicological impact of parent drug versus phase I metabolites of synthetic cannabinoids on human cells with and without CB1 receptor. The human cell line of neuroblastoma SH-SY5Y and human kidney cell line HEK-293T were exposed to JWH-018 and to its N-(3-hydroxypentyl) metabolite. Cell toxicity was evaluated using the MTT and LDH assay. Additionally, a dual staining methodology with fluorescent Annexin V-FITC and propidium iodide was performed to address the question of whether JWH-018 N-(3-hydroxypentyl) metabolite is inducing cell death through apoptosis or necrosis, in HEK293T and SH-SY5Y cell lines. The obtained results show that JWH-018 does not cause a statistically significant decrease in cell viability, in contrast to its N-(3-hydroxypentyl) metabolite, which at ≥25μM causes a significant decrease in cell viability. Both cell lines are affected by JWH-018 metabolite. Our results point to higher toxicity of JWH-018 metabolite when compared to its parent drug, suggesting a non-CB1 receptor mediated toxicological mechanism. Comparing the results from Annexin V/PI with MTT and LDH assays of SH-SY5Y and HEK293T in the presence of the synthetic cannabinoid metabolite, emerges the picture that cellular viability decreases and associated death is occurring through necrosis."

Leishmanicidal and cytotoxic activity of extracts and saponins from Ilex laurina (Aquifoliaceae)

Purpose: To evaluate the leishmanicidal and cytotoxic activity of alcohol and non-alcohol extracts and saponins from Ilex laurina . Methods: Extracts were obtained by percolation with solvents of different polarities: hexane, dichloromethane, ethyl acetate and ethanol. The ethyl acetate extract was subjected to silica gel column chromatography eluting with a step gradient of dichloromethane-methanol. All products were evaluated in vitro for leishmanicidal activity against amastigotes of leishmania panamensis and cytotoxicity on U- 937 cells. Results: Two saponins were isolated from the ethyl acetate extract. The ethyl acetate extract showed high leishmanicidal activity against intracellular amastigotes of L. panamensis (EC50, 7.5 ± 1.5 μg/mL) and low activity against axenic amastigotes (EC50, 52.8 ±1.6 μg/mL); this extract showed also high cytotoxicity (LC50, 57.7 ± 12.1 μg/mL). Saponin 2 exhibited high activity against intracellular amastigotes (EC50, 5.9 ± 0.5 μg/mL) but also showed high cytotoxicity on U-937 cells (EC50, 25.7 ± 6.1 μg/mL). This compound showed similar leishmanicidal activity and cytotoxicity to meglumine antimoniate and amphotericin B, respectively, drugs currently used for the treatment of leishmaniasis. Conclusions: Based on these results, Ilex laurina is a potential source of compounds that can lead to the development of new therapeutic alternatives against leishmaniasis.