Synergistic In Vitro Antiviral Effect of Combinations of Ivermectin, Essential Oils, and 18-(Phthalimid-2-yl)ferruginol against Arboviruses and Herpesvirus.

Combining antiviral drugs with different mechanisms of action can help prevent the development of resistance by attacking the infectious agent through multiple pathways. Additionally, by using faster and more economical screening methods, effective synergistic drug candidates can be rapidly identified, facilitating faster paths to clinical testing. In this work, a rapid method was standardized to identify possible synergisms from drug combinations. We analyzed the possible reduction in the antiviral effective concentration of drugs already approved by the FDA, such as ivermectin (IVM), ribavirin (RIBA), and acyclovir (ACV) against Zika virus (ZIKV), Chikungunya virus (CHIKV), and herpes virus type 2 (HHV-2). Essential oils (EOs) were also included in the study since they have been reported for more than a couple of decades to have broad-spectrum antiviral activity. We also continued studying the antiviral properties of one of our patented molecules with broad-spectrum antiviral activity, the ferruginol analog 18-(phthalimid-2-yl)ferruginol (phthFGL), which presented an IC99 of 25.6 µM for the three types of virus. In general, the combination of IVM, phthFGL, and oregano EO showed the greatest synergism potential against CHIKV, ZIKV, and HHV-2. For instance, this combination achieved reductions in the IC99 value of each component up to ~8-, ~27-, and ~12-fold for CHIKV, respectively. The ternary combination of RIBA, phthFGL, and oregano EO was slightly more efficient than the binary combination RIBA/phthFGL but much less efficient than IVM, phthFGL, and oregano EO, which indicates that IVM could contribute more to the differentiation of cell targets (for example via the inhibition of the host heterodimeric importin IMP α/ß1 complex) than ribavirin. Statistical analysis showed significant differences among the combination groups tested, especially in the HHV-2 and CHIKV models, with p = 0.0098. Additionally, phthFGL showed a good pharmacokinetic profile that should encourage future optimization studies.

A new host-targeted antiviral cyclolignan (SAU-22.107) for Dengue Virus infection in cell cultures. Potential action mechanisms based on cell imaging.

Dengue virus (DENV) infection is the most arbovirosis in the world. However, medications have not been approved for its treatment. Drug discovery based on the host-targeted antiviral (HTA) constitutes a new promising strategy, considering their high genetic barrier to resistance and the low probability of selecting drug resistance strains. In this study, we have tested fifty-seven podophyllotoxin-related cyclolignans on DENV-2 infected cells and found the most promising compound was S.71. Using cellular and molecular biology experiments, we have discovered that the new lignan altered the distribution of microtubules, induced changes in cell morphology, and caused retraction of the rough endoplasmic reticulum. In addition, the compound alters the viral envelope protein and the double-stranded RNA, while there is a decrease in negative-strand RNA synthesis; especially when the compound was added between 6- and 12-hours post-infection. Altogether, S.71 decreases the viral yield through an HTA-related mechanism of action, possibly altering the DENV genome replication and/or polyprotein translation, through the alteration of microtubule distribution and endoplasmic reticulum deterioration. Finally, pharmacokinetic predictors show that S.71 falls within the standard ranges established for drugs.

The Colombian Zika Virus Isolate (COL345Si) Replicates in Prostate Adenocarcinoma Cells and Modulates the Antiviral Response.

Zika virus (ZIKV), a flavivirus that is mainly transmitted by A. aegypti and A. albopictus and sexual transmission, has been documented and described. The ZIKV RNA detection in the semen of vasectomized men indicates that accessory glands such as the prostate could be a site of virus replication. In this study, we characterized the ZIKV infection, evaluated the antiviral profile, and demonstrated the AXL and TIM-1 expression on the PC3 prostate cell line. It was also determined that PC3 cells are susceptible and permissive to ZIKV infection without altering the cell viability or causing a cytopathic effect. The antiviral profile suggests that the PC3 cells modulate the antiviral response through the suppressor molecule expression, SOCS-1, during a ZIKV infection.

Semisynthetic Abietic and Dehydroabietic Acid Derivatives and Triptoquinone Epimers Interfere with LPS-Triggered Activation of Dendritic Cells.

Abietic acid (AA), dehydroabietic acid (DHA) and triptoquinones (TQs) are bioactive abietane-type diterpenoids, which are present in many edible vegetables and medicinal herbs with health-promoting properties. Evidence suggests that beneficial effects of diterpenes operate, at least in part, through effects on cells in the immune system. Dendritic cells (DCs) are a key type of leukocyte involved in the initiation and regulation of the immune/inflammatory response and natural or synthetic compounds that modulate DC functions could be potential anti-inflammatory/immunomodulatory agents. Herein, we report the screening of 23 known semisynthetic AA and DHA derivatives, and TQs, synthesized previously by us, in a multi-analyte DC-based assay that detects inhibition of pro-inflammatory cytokine production. Based on the magnitude of the inhibitory effect observed and the number of cytokines inhibited, a variety of activities among compounds were observed, ranging from inactive/weak to very potent inhibitors. Structurally, either alcohol or methyl ester substituents on ring A along with the introduction of aromaticity and oxidation in ring C in the abietane skeleton were found in compounds with higher inhibitory properties. Two DHA derivatives and two TQs exhibited a significant inhibition in all pro-inflammatory cytokines tested and were further investigated. The results confirmed their ability to inhibit, dose dependently, LPS-stimulated expression of the co-stimulatory molecules CD40 and/or CD86 and the production of the pro-inflammatory cytokines IL-1ß, IL-6, IL-12 and TNFα. Our results demonstrate that DC maturation process can be targeted by semisynthetic DHA derivatives and TQ epimers and indicate the potential of these compounds as optimizable anti-inflammatory/immunomodulatory agents.

Antiviral Profiling of C-18- or C-19-Functionalized Semisynthetic Abietane Diterpenoids.

Viral infections affect several million patients annually. Although hundreds of viruses are known to be pathogenic, only a few can be treated in the clinic with available antiviral drugs. Naturally based pharmacotherapy may be a proper alternative for treating viral diseases. Several natural and semisynthetic abietane-type diterpenoids have shown important antiviral activities. In this study, a biological evaluation of a number of either C-18- or C-19-functionalized known semisynthetic abietanes against Zika virus, Dengue virus, Herpes virus simplex type 1, and Chikungunya virus are reported. Semisynthetic abietane ferruginol and its analogue 18-(phthalimid-2-yl)ferruginol displayed broad-spectrum antiviral properties. The scale-up synthesis of this analogue has been optimized for further studies and development. This molecule displayed an EC50 between 5.0 and 10.0 µM against Colombian Zika virus strains and EC50 = 9.8 µM against Chikungunya virus. Knowing that this ferruginol analogue is also active against Dengue virus type 2 (EC50 = 1.4 µM, DENV-2), we can conclude that this compound is a promising broad-spectrum antiviral agent paving the way for the development of novel antivirals.

Anti-Herpetic, Anti-Dengue and Antineoplastic Activities of Simple and Heterocycle-Fused Derivatives of Terpenyl-1,4-Naphthoquinone and 1,4-Anthraquinone.

Quinones are secondary metabolites of higher plants associated with many biological activities, including antiviral effects and cytotoxicity. In this study, the anti-herpetic and anti-dengue evaluation of 27 terpenyl-1,4-naphthoquinone (NQ), 1,4-anthraquinone (AQ) and heterocycle-fused quinone (HetQ) derivatives was done in vitro against Human Herpesvirus (HHV) type 1 and 2, and Dengue virus serotype 2 (DENV-2). The cytotoxicity on HeLa and Jurkat tumor cell lines was also tested. Using plaque forming unit assays, cell viability assays and molecular docking, we found that NQ 4 was the best antiviral compound, while AQ 11 was the most active and selective molecule on the tested tumor cells. NQ 4 showed a fair antiviral activity against Herpesviruses (EC50: <0.4 µg/mL, <1.28 µM) and DENV-2 (1.6 µg/mL, 5.1 µM) on pre-infective stages. Additionally, NQ 4 disrupted the viral attachment of HHV-1 to Vero cells (EC50: 0.12 µg/mL, 0.38 µM) with a very high selectivity index (SI = 1728). The in silico analysis predicted that this quinone could bind to the prefusion form of the E glycoprotein of DENV-2. These findings demonstrate that NQ 4 is a potent and highly selective antiviral compound, while suggesting its ability to prevent Herpes and Dengue infections. Additionally, AQ 11 can be considered of interest as a leader for the design of new anticancer agents.

Anti-herpetic and anti-dengue activity of abietane ferruginol analogues synthesized from (+)-dehydroabietylamine.

The abietane-type diterpenoid (+)-ferruginol (1), a bioactive compound isolated from several plants, has attracted much attention as consequence of its pharmacological properties, which includes antibacterial, antifungal, antimicrobial, cardioprotective, anti-oxidative, anti-plasmodial, leishmanicidal, anti-ulcerogenic, anti-inflammatory and antitumor actions. In this study, we report on the antiviral evaluation of ferruginol (1) and several analogues synthesized from commercial (+)-dehydroabietylamine. Thus, the activity against Human Herpesvirus type 1, Human Herpesvirus type 2 and Dengue Virus type 2, was studied. Two ferruginol analogues showed high antiviral selectivity index and reduced viral plaque-size in post-infection stages against both Herpes and Dengue viruses. A promising lead, compound 8, was ten-fold more potent (EC50 = 1.4 µM) than the control ribavirin against Dengue Virus type 2. Our findings suggest that the 12-hydroxyabieta-8,11,13-triene skeleton, which is characteristic of the diterpenoid ferruginol (1), is an interesting molecular scaffold for development of novel antivirals. In addition, the cytotoxic and antifungal activities of the synthesized ferruginol analogues have also been investigated. (©)20155 Elsevier Science. All rights reserved.

Synthesis and antifungal activity of terpenyl-1,4-naphthoquinone and 1,4-anthracenedione derivatives.

The antifungal evaluation of twenty seven simple and heterocycle-fused prenyl-1,4-naphthoquinones and 1,4-anthracenediones was performed in vitro against human pathogenic yeasts (Candida spp.) and filamentous fungi (Aspergillus spp., Fusarium spp., and Trichophyton spp.). The synthetic strategy used to obtain the quinone derivatives was initially based on the Diels-Alder cycloaddition between myrcene and several p-benzoquinone derivatives, followed by cyclisation of the prenyl side chain in the case of anthracene-1,4-diones. The most promising compounds, displaying MIC values in the low µg/mL range, were those bearing one or two chlorine atoms attached to the quinone ring. Time-kill curves determined for the most potent compounds showed their fungistatic mode of action similar to that of itraconazole.

Synthesis and biological evaluation of combretastatin A-4 and three combretastatin-based hybrids.

The syntheses of combretastatin A-4 from gallic acid and of three combretastatin-based hybrids are described. Starting from commercial gallic acid, the phosphonium salt (3,4,5-trimethoxybenzylphosphonium bromide) is synthesized and coupled, through a Wittig reaction, with several aldehydes, including methoxymethyl-protected isovanillin, the aldehyde y-bicyclohomofarnesal having a labdane skeleton, 3-(3-pyridyl) propanal, and furfural. The biological properties of the cis-coupled compounds as cytotoxic, antiviral and antifungal agents are also reported. In addition, pyrogallol, gallic and 3,4,5-trimethoxybenzoic acids have been studied biologically.

Enhanced dissipation of polycyclic aromatic hydrocarbons in the rhizosphere of the Athel tamarisk (Tamarix aphylla L. Karst.) grown in saline-alkaline soils of the former lake Texcoco.

Remediation of polycyclic aromatic hydrocarbons (PAHs) contaminated alkaline saline soil with phreatophyte or "water loving plants" was investigated by spiking soil from the former lake Texcoco with 100 mg phenanthrene (Phen) kg(-1) soil, 120 mg anthracene (Ant)kg(-1) soil and 45 mg benzo(a)pyrene (BaP) kg(-1) soil and vegetating it with Athel tamarisk (Tamarix aphylla L Karst.). The growth of the Athel tamarisk was not affected by the PAHs. In soil cultivated with Athel tamarisk, the leaching of PAHs to the 32-34 cm layer decreased 2-fold compared to the uncultivated soil. The BaP concentration decreased to 39% of the initial concentration at a distance smaller than 3 cm from the roots and to 45% at a distance larger than 3cm, but 59% remained in unvegetated soil after 240 days. Dissipation of Ant and Phen decreased with depth, but not BaP. The biodegradation of PAHs was affected by their chemical properties and increased in the presence of T. aphylla, but decreased with depth.

Antifungal, cytotoxic and chemical analyses of essential oils of Lippia origanoides HBK grown in Colombia / Actividad antifúngica, citotóxica y composición química de aceites esenciales de Lippia origanoides HBK recolectadas en Colombia

Introduction: Aspergillus fumigatus is most commonly associated to invasive aspergillosis. Strong antifungal activity against A. fumigatus of L. origanoides essential oil gives a new added value to this natural product from Boyacá-Colombia. Aims: The increase in fungal infections, the development of resistance and toxicity of wide-spectrum antifungals have led to a constant search for therapeutic alternatives. The chemical composition, antifungal and cytotoxic activity of nine essential oils obtained from L. origanoides were evaluated and the relationship between the antifungal activities of the oil and of its major components were explored. Methods and Results: Antifungal activity was determined following the protocols AFST-EUCAST for Candida krusei and C. parapsilosis, and CLSI-M38A for Aspergillus fumigatus and A. flavus. The GC-MS analysis identified three chemotypes: thymol, carvacrol and p-cymene/trans-beta-caryophyllene. The essential oil of the thymol chemotype was the most active in antifungal assays with MIC values of 157.5, 198.4, 125 and 31 μg ml-1 against C. parapsilosis, C. krusei, A. flavus and A. fumigatus, respectively. The major components carvacrol and thymol were not active against A. fumigatus at concentrations below 157.5 μg ml-1. In general, the oils were not cytotoxic. Conclusions: The essential oil of the thymol chemotype of L. origanoides from the region of Boyacá- Colombia showed the highest antifungal activity against A. fumigatus among all the oils and major components tested. Salud UIS 2011; 43 (2): 141-148.

Introducción: La infección por el hongo Aspergillus fumigatus está más comúnmente asociada a la aspergilosis invasiva. La fuerte actividad antimicótica del aceite esencial de L. origanoides contra A. fumigatus ha dado un nuevo valor agregado a este producto natural de Boyacá-Colombia. Objetivo: El aumento de las infecciones por hongos, el desarrollo de la resistencia y la toxicidad de los antifúngicos de amplio espectro han llevado a una constante búsqueda de alternativas terapéuticas. En este estudio fueron evaluados la composición química, la actividad antifúngica y citotóxica de nueve aceites esenciales obtenidos de L. origanoides; y la relación entre la actividad antifúngica de los aceites con respecto a la presencia de sus principales componentes. Métodos y Resultados: La actividad antifúngica se determinó siguiendo los protocolos AFST-EUCAST para Candida krusei y C. parapsilosis; y CLSI M38A para Aspergillus fumigatus y A. flavus. El análisis por GC-MS identificó tres quimiotipos: carvacrol timol y p-cymene/trans-beta-caryophyllene. El aceite esencial del quimiotipo timol fue el más activo en los ensayos antifúngicos con valores de MIC de 157,5, 198,4, 125 y 31 mg ml-1 frente a C. parapsilosis, C. krusei, A. flavus y A. fumigatus, respectivamente. El carvacrol y el timol, los principales componentes, no fueron activos frente a A. fumigatus en concentraciones inferiores a 157,5 g / ml-1. En general, los aceites no fueron citotóxicos. Conclusiones: El aceite esencial de L origanoides, quimiotipo timol, de la región de Boyacá-Colombia presentó la mayor actividad antifúngica frente a A. fumigatus entre todos los aceites evaluados; igualmente, sus principales componentes fueron los más activos en comparación a los otros quimiotipos. Salud UIS 2011; 43 (2): 141-148.

Composicion quimica y evaluacion de la actividad antiherpetica in vitro de aceites esenciales de Lippia alba (Mill) N.E. Brown y sus componentes mayoritarios / Chemical Composition and evaluation in vitro of anti-herpetic activity of essential oils from Lippia alba (Mill) NE Brown and the main components

Introduccion: Lippia alba (Mill) N.E. Brown es una planta aromatica perteneciente a la familia Verbenaceae, ampliamente usada en Suramerica y Norteamerica como infusion contra la hipertension, problemas digestivos, nauseas y resfriados. Objetivo: En el presente estudio, se evaluo la actividad antiviral in vitro frente al Herpesvirus Humano Tipo 1 (HSV-1), de veinte aceites esenciales de L. alba y, diez de sus componentes mayoritarios. Metodologia: La actividad antiviral in vitro fue evaluada empleando la t¨¦cnica modificada, de titulacion del punto final (EPTT). Los aceites esenciales, de plantas de L. alba recolectadas de diferentes regiones del pais, fueron obtenidos por hidrodestilacion asistida con radiacion de microondas (MWHD). Se determino por cromatografia de gases¨Cespectrometria de masas (GC-MS), la composicion quimica, de los veinte aceites esenciales de L. alba, identificandose dos quimiotipos: citral y carvona. Resultados y conclusiones: Los aceites quimiotipo carvona, BC1 y CA2, mostraron actividad antiherpetica in vitro, moderada sobre monocapa de celulas HeLa infectadas, con valores de Rf de 1x101,5, en concentraciones, de 250¦Ìg/mL y 125¦Ìg/mL, respectivamente. Los controles positivos, sulfato de heparina y aciclovir, redujeron el titulo viral con valores de Rf, en orden, de 1x102 y 1x104. Ninguno de los monoterpenos evaluados mostro actividad contra el HSV-1.

Introduction: Lippia alba (Mill.) N.E. Brown is an aromatic shrub belonging to the Verbenaceae family, which is widely used all over South and Central America as an infusion against, hypertension, digestive troubles, nausea and cold. The antiviral activity in vitro against Herpesvirus 1, Human (HSV-1) of twenty essential oils of L. alba and ten of its components were evaluated. Methodology: The antiviral activity was determined using a modified end-point titration technique (EPPT). The essential oils from L. alba collected in different regions of Colombia were obtained by microwave-assisted hydrodistillation (MWHD). Their chemical compositions were determined by GC and GC/MS. Two chemotypes were distinguished, characterized by carvone and citral as main constituents. Results and conclusions: Carvone chemotype oils BC1 and CA2 were found to be moderate antiviral activity over infected confluent monolayers of HeLa cells with Rf value of 1x101,5 at a concentration of 250¦Ìg/mL and 125¦Ìg/mL, respectively. Heparin sodium salt and acyclovir were used as positive controls and showed Rf values of 1x102 and 1x104, respectively. None of monoterpenes tested showed antiviral activity against HSV-1.

Synthesis and biological evaluation of (+)-labdadienedial, derivatives and precursors from (+)-sclareolide.

Labdadienedial and a series of C15,C16-functionalized derivatives were synthesized from commercial (+)-sclareolide and evaluated for their cytotoxic, antimycotic, and antiviral activities. Their precursors were similarly evaluated.

[Antifungal activity, cytotoxicity and composition of essential oils from the Asteraceae plant family]. / Actividad antimicótica y citotóxica de aceites esenciales de plantas de la familia Asteraceae.

BACKGROUND: The plants of the Asteraceae family have been used for medicinal purposes,in traditional Colombian medicine. AIM: To evaluate the antifungal activity and the cytotoxic effects of 15 essential oils from plants of the Asteraceae family. METHODS: Antifungal activity was evaluated against Candida parapsilosis ATCC 22019, Candida krusei ATCC 6258, Aspergillus flavus ATCC 204304 and Aspergillus fumigatus ATCC 204305 following EUCAST and CLSI M38-A standard methods, for yeast and filamentous fungi, respectively. Cytotoxic effect was evaluated on Vero cell line by MTT assay. RESULTS: The oils from the plants Achyrocline alata and Baccharislatifolia were the only ones active against A. fumigatus (GM-MIC=78.7 and 157.4 microg/ml, respectively). In contrast, there was no evidence of oils active against Candida species. In addition, these oils were not cytotoxic on Vero cells. The oils of A. alata and Baccharis latifolia could be candidates for disinfecting hospital environments and for inhibiting biofilm formation by A. fumigatus CONCLUSIONS: The oils of A. alata and B. latifolia could be candidates for disinfecting hospital environments and for inhibiting biofilm formation by A. fumigatus.

Synthesis and antiviral activity of scopadulane-rearranged diterpenes.

A new bioactive diterpene skeleton resulting from a backbone rearrangement is described. Activity of the rearranged product and several derivatives against Herpes Virus Simplex type 2 is reported.

Synthesis and biological evaluation of dehydroabietic acid derivatives.

A series of C18-oxygenated derivatives of dehydroabietic acid were synthesized from commercial abietic acid and evaluated for their cytotoxic, antimycotic, and antiviral activities.

Actividad citotóxica de aceites esenciales de Lippia origanoides H.B.K. y componentes mayoritarios / Cytotoxic activity of essencial oils of Lippia origanoides H.B.K and its major constituents

Introducción: Lippia origanoides H.B.K. (Verbenacea), es una planta aromática conocida comúnmente como “orégano”. Los aceites esenciales de 8 muestras de L. origanoides y algunos de sus componentes mayoritarios fueron evaluados in vitro sobre la línea tumoral HeLa y la línea no tumoral Vero para identificar su potencial citotóxico. Materiales y métodos: la concentración inhibitoria cincuenta (IC50) se determinó mediante la técnica fotocolorimétrica del MTT (3-(4,5-Dimetiltiazol-2-il)-2,5-bromuro difeniltetrazolio) y los valores de IC50 se obtuvieron por análisis estadístico mediante regresión lineal simple. El índice de selectividad (IS), definido como la IC50 en células Vero sobre IC50 en células HeLa, fue calculado con el fin de encontrar aceites o componentes con potencial citotóxico selectivo hacia líneas celulares tumorales. Resultados y conclusiones: se determinó por cromatografía de gases y espectrometría de masas GC/MS la composición química de los aceites más citotóxicos. El aceite de L. origanoides que presentó la mayor actividad citotóxica sobre células HeLa con un valor de IC50 de 9.1 ± 1 µg/mL e índice de selectividad de 7,1, fue identificado como quimiotipo trans-β-cariofileno/r-cimeno. Los componentes mayoritarios del aceite quimiotipo trans-β-cariofileno/r-cimeno fueron: trans-β-cariofileno (11.3%), r-cimeno (11,2%), α-felandreno (9,9%), limoneno (7,2%), 1,8-cineol (6,5%) y α-humuleno (6,0%). Los componentes mayoritarios evaluados no mostraron actividad citotóxica relevante sobre células HeLa, sólo el limoneno y β-mirceno presentaron valores de IS, respectivamente, de 6,97 y 3,01. Sin embargo, los valores de IC50 fueron más altos que el del aceite activo. Estos resultados sugieren que la actividad citotóxica de los aceites no se debe sólo a sus componentes mayoritarios, sino a un sinergismo entre sus componentes.

Introduction: Lippia origanoides H.B.K. (Verbenaceae) is an aromatic plant commonly called as "oregano". Eight essential oils of L. origanoides and some of their main components were evaluated in vitro on tumor cell line HeLa and non-tumor cell line Vero to identify tumoural cytotoxic potential. Materials and methods: Inhibition 50% of cell population (IC50) was determined using the photo-colorimeter technique MTT (3 - (4.5-dimethylthiazol-2-yl) -2,5-difeniltetrazolium bromide). IC50 values were obtained by linear regression analysis. The selectivity index (SI), defined as Vero IC50 on HeLa IC50, it was calculated in order to find oil or major components with selective tumor cytotoxic potential. Results and conclusions: the chemical composition of the oil most cytotoxic was determined by gas chromatography and mass spectrometry GC/MS. The L. origanoides oil identified as chemotype trans-β- caryophyllene/p-cymene showed the highest cytotoxic activity on HeLa cells with IC50 value of 9.1 ± 1 µg/mL and selectivity index of 7.1. The main components were: trans-β-caryophyllene (11.3 p-cymene (11.2%), α-phellandrene (9.9%), limonene (7.2%), 1.8-cineol (6.5%) and α-humulene (6.0%). The most of the major components did not show cytotoxic activity on HeLa cells, only limonene and β-myrcene showed IS values of 6.97 and 3.01, respectively. However, the IC50 values were higher than active oil. These results suggest that cytotoxic activities of the oils are not only due to their main components, but to a synergism among its components.

Actividad antimicótica, citotoxicidad y composición de aceites esenciales de plantas de la familia Labiatae / Antifungal activity, cytotoxicity and composition of essential oils from Asteraceae family plants

Introducción: Candida spp. y Aspergillus spp. son causa importante de infecciones a nivel mundial. Considerando la resistencia de estos patógenos a algunos de los antimicóticos disponibles, es necesaria la búsqueda de nuevos agentes antimicóticos. Diferentes aceites esenciales y extractos de plantas han mostrado actividad antimicótica in vitro. El objetivo de este estudio fue evaluar la actividad antimicótica, citotóxica y la composición química de aceites esenciales de la familia Labiatae. Materiales y métodos: Se evaluó la actividad antimicótica de 22 aceites de plantas de la familia Labiatae contra C. parapsilosis ATCC 22019, C. krusei ATCC 6258, A. flavus ATCC 204304 y A. fumigatus ATCC 204305, siguiendo las técnicas estándar EUCAST y CLSI M38-A para levaduras y hongos filamentosos, respectivamente. Adicionalmente la actividad citotóxica se evaluó en la línea celular Vero mediante la técnica colorimétrica del MTT. La caracterización de los aceites esenciales se llevó a cabo por cromatografía de gases acoplada a masas. Resultados: El aceite esencial mas activo fue el de Minthostachys mollis frente a todas las cepas evaluadas con rangos concentraciones mínimas inhibitorias (CMIs) entre 250 y 375 µg/mL. El aceite de la planta Hyptis mutabilis mostró actividad frente a A. fumigatus (CMI = 396.8 µg/mL). Estos aceites esenciales no fueron citotóxicos sobre las células Vero. Los componentes principales de los aceites de las plantas M. mollis y H. mutabillis fueron epóxido de cis-piperitona y 1,8-cineol, respectivamente. Conclusiones: Los aceites esenciales de las plantas M. mollis y H. mutabillis mostraron actividad antimicótica y no fueron citotóxicos en células Vero.

Introduction: Aspergillus spp. and Candida spp. are important cause of infections worldwide. Considering the resistance of these pathogens to some antifungal agents, there is greater need to search for new antifungal agents. Many extracts and essential oils isolated from plants have shown to exert antifungal effects in vitro. The aim of this study was to evaluate the antifungal, cytotoxic effect, and chemical composition of essential oils of family Labiatae. Materials and methods: Antifungal activity of twenty two essential oils from Labiatae family was evaluated against C. parapsilosis ATCC 22019, C. krusei ATCC 6258, A. flavus ATCC 204304 y A. fumigatus ATCC 204305, following EUCAST and M38-A standard protocols for yeast and filamentous fungi, respectively. Additionally, cytotoxic activity was evaluated on Vero cell line by colorimetric assay MTT. Essential oils was characterized by gas chromatography coupled to mass spectrometry. Results: The most active oil with all strains was obtained of Minthostachys mollis (MIC range 250 - 375 µg/mL). The essential oil from Hyptis mutabillis showed activity against A. fumigatus (GM-MIC = 396.8 µg/mL). These essential oils were not cytotoxic on Vero cells. The major components of essential oils from M. mollis and H. mutabillis were cis-piperitone epoxide and 1,8-cineol, respectively. Conclusions: Essential oils of H. mutabillis and M. mollis showed antifungal activity and they were not cytotoxic on Vero cells.

Composición química y actividad biológica de aceites esenciales de Calycolpus moritzianus recolectado en el Norte de Santander, Colombia / Chemical composition and biological activity of essential oils calycolpus moritzianus collectd in the Norte de Santander, Colombia

Introducción: actualmente, se están llevando a cabo estudios sobre la actividad biológica de AEs de plantas de la familia Myrtaceae. El objetivo de esta investigación fue la caracterización química y la evaluación de la actividad antimicótica, antitumoral y antiviral de 40 aceites extraídos de plantas de la especie Calycolpus moritzianus, comúnmente llamada “arrayán ó cínaro”, recolectadas en varias zonas del Norte de Santander, Colombia. Materiales y métodos: la actividad antimicótica se evaluó contra: Candida krusei, C. parapsilosis, Aspergillus fumigatus y A. flavus, siguiendo las técnicas estándar EUCAST y CLSI M38-A, respectivamente, para levaduras y hongos filamentosos. La citotoxicidad se evaluó en las líneas celulares tumorales (HeLa, Jurkat) y no tumorales (Vero) mediante la técnica fotocolorimétrica del MTT. La actividad antiviral in vitro, contra el virus Herpes simplex tipo HSV-1, se realizó mediante la “técnica de titulación del punto final (EPTT)”. Resultados y Discusión: los aceites X38 y X39, de muestras recolectadas en los municipios de Chinácota y Salazar, mostraron actividad contra C. krusei con valores de concentraciones inhibitorias mínimas de 500 µg/mL. Ninguno de los aceites fue citotóxico en células no tumorales Vero. Los aceites X9 y X10, procedentes de los municipios de Salazar y Pamplonita, fueron los más citotóxicos sobre células HeLa, a concentraciones de 37,8 ± 2.4 y 40,6 ± 2,6 µg/mL, respectivamente. La reducción de la carga viral en los cultivos celulares sólo fue lograda por el aceite X11. La actividad antimicótica y antitumoral de los aceites fue correlacionada con la composición de los componentes mayoritarios, evidenciándose la actividad del Terpinen-4-ol, (-) Limoneno, α –Pineno y Linalol.

Background: Essential oils of plants of the family Myrtaceae have been evaluated for different biological activities. The aim of this research was the chemical characterization and evaluation of antifungal, antiviral and cytotoxic activity of 40 essential oils extracted from Calycolpus moritzianus, commonly called “arrayán” or “cínaro”, collected in several areas of Norte de Santander, Colombia. Methods: antifungal activity was determined following the protocols AFST-EUCAST for Candida krusei and C. parasilopsis, and CLSI-M38A for Aspergillus fumigatus and A. flavus. Cytotoxicity was evaluated on tumoral (HeLa and Jurkat), and non tumoral (Vero) cell lines by the MTT photo-colorimetric technique The in vitro antiviral activity against Herpes simplex virus HSV-1 was determined by the end-point titration technique (EPTT). Results and Discussion: X38 and X39 oils of plants collected in regions of Chinácota and Salazar regions showed activity against C. krusei with minimal inhibitory concentration values of 500 µg/mL. None of the oils was cytotoxic in no-tumor (Vero) cells . Oils X9 and X10 of plants from regions of Salazar and Pamplonita were the most cytotoxic on HeLa cells at concentrations of 37,8 ± 2,4 and 40,6 ± 2,6 µg /mL, respectively. The reduction of viral load in cell cultures only was showed by X11 oil. Antifungal and antitumor activity of these oils was correlated with the composition of the major components showing activity for Terpinen-4-ol, (-) Limonene, Linalool and α-Pinene.

Citral and carvone chemotypes from the essential oils of Colombian Lippia alba (Mill.) N.E. Brown: composition, cytotoxicity and antifungal activity.

Two essential oils of Lippia alba (Mill.) N.E. Brown (Verbenacea), the carvone and citral chemotypes and 15 of their compounds were evaluated to determine cytotoxicity and antifungal activity. Cytotoxicity assays for both the citral and carvone chemotypes were carried out with tetrazolium-dye, which showed a dose-dependent cytotoxic effect against HeLa cells. Interestingly, this effect on the evaluated cells (HeLa and the non-tumoural cell line, Vero) was lower than that of commercial citral alone. Commercial citral showed the highest cytotoxic activity on HeLa cells. The antifungal activity was evaluated against Candida parapsilosis, Candida krusei, Aspergillus flavus and Aspergillus fumigatus strains following the standard protocols, Antifungal Susceptibility Testing Subcommittee of the European Committee on Antibiotic Susceptibility Testing and CLSI M38-A. Results demonstrated that the most active essential oil was the citral chemotype, with geometric means-minimal inhibitory concentration (GM-MIC) values of 78.7 and 270.8 microg/mL for A. fumigatus and C. krusei, respectively. Commercial citral showed an antifungal activity similar to that of the citral chemotype (GM-MIC values of 62.5 microg/mL for A. fumigatus and 39.7 microg/mL for C. krusei). Although the citronellal and geraniol were found in lower concentrations in the citral chemotype, they had significant antifungal activity, with GM-MIC values of 49.6 microg/mL for C. krusei and 176.8 microg/mL for A. fumigatus.