Preclinical pharmacokinetics of a promising antineoplastic prototype piperazine-containing compound (LQFM018) in rats by a new LC-MS/MS bioanalytical method.

LQFM018 is a novel antineoplastic prototype, showing an expressive drug-triggered K562 leukemic cells death mechanism, through necroptotic signaling. Due to its promising effect, this study aimed to evaluate the pharmacokinetics of LQFM018 in rats, using a new validated bioanalytical LC-MS/MS-based method. Chromatographic column was an ACE® C18 (100 mm × 4.6 mm, 5 µm) eluted by a mobile phase composed of ammonium acetate 2 mM and formic acid 0.025%:methanol (50:50, v/v), under flow of 1.2 mL/min and injection volume of 3.0 µL. LQFM018 was extracted from rat plasma by a simple liquid-liquid method, using MTBE solvent. Rats were administered intraperitoneally at LQFM018 100 mg/kg dose and blood samples were collect at times of 0, 1, 2, 3, 4, 5, 6, 7, 8, and 9 h. Bioanalytical-LC-MS/MS-based method was rapid, high throughput and sensitive with a good linearity ranging from 10 (LLOQ) to 15000 ng/mL, besides precise and accurate, ranging of 0.8-7.3% and 96.8-107.6%, respectively. The prototype LQFM018 was rapid and well absorbed, and highly distributed, apparently due to its high lipid solubility. These features are primordial for an anticancer agent in the treatment of deep tumors, such as bone marrow neoplasms, in which the drug might permeate easily tissue barriers. Also, LQFM018 has demonstrated a high clearance, according to a low t1/2in rats, indicating a relative fast elimination phase related to a possible intense hepatic biotransformation. These information support further studies to establish new understands on pharmacokinetics of promising antineoplastic prototype LQFM018 from preclinical and clinical evaluations.

Pro-adherence complementary audiovisual educational intervention model for chronic myeloid leukemia patients treated with imatinib mesylate.

BACKGROUND: Imatinib mesylate (IM) treatment adherence is a challenge, especially in an economic-social population neglected from developing countries. OBJECTIVE: To create a new complementary audiovisual educational intervention model to improve IM treatment adherence of CML patients. METHODS: Two adherence verification methods were applied before and after intervention: modified Morisky-Green test and molecular responses (BCR-ABL transcripts quantification). Adherence estimates were calculated using univariate and multivariate component analysis (MCA) for the socio-demographic and clinical characteristics of patients. RESULTS: Modified Morisky-Green test results demonstrated a substantial increase of CML patient adherence from 23% (pre-film) to 65% (post-film). Greater improvement was obtained for patients presenting major molecular response (MMR) from 38% (pre-film) to 60% (post-film). Although slight gain for complete molecular response (CMR) from 23% (pre-film) to 26% (post-film) was achieved, it represents a total tumour regression. MCA identified that females <50 years-old, using less than two medications (no disease associated) and CMR condition were the most benefited with intervention. CONCLUSION: Audiovisual educational intervention was an effective complementary pro-adherence model, activating patient memory and improving IM treatment adherence. Although this intervention shows effective, not all patients responded as expected, being necessary a combination of educational and clinical interventions to improve IM adherence.

Antihypertensive effect of Equisetum arvense L.: a double-blind, randomized efficacy and safety clinical trial.

BACKGROUND: Equisetum arvense L. (EA) is a traditional phytomedicine used as a diuretic agent worldwide and regulated strictly by European Medicine Agency (EMA) and Brazilian National Health Surveillance Agency (ANVISA). However, few studies evaluating its efficacy and safety have been published and no clinical trial assessing its antihypertensive effect has been reported to date. PURPOSE: To assess antihypertensive effect, safety and tolerability of EA compared to hydrochlorothiazide (HCTZ). METHODS: This is a double-blind randomized clinical trial, allocating 58 systemic arterial hypertension (SAH) stage I patients (both sexes, 25-65 years old) into two groups (EA and HCTZ). All patients underwent biochemical and cardiologic checkup prior to and during interventions. The EA standardized dry extract (900 mg/day) or HCTZ (25 mg/day) were administered for 3 months and follow-up visits were conducted every 30 days. Efficacy established goals were systolic blood pressure (SBP) and/or diastolic blood pressure (DBP) decreases ≥ 10.0 mmHg and/or casual blood pressure (CBP) < 140/90 mmHg. RESULTS: EA treatment demonstrated a significant antihypertensive effect, promoting a mean decrease of SBP and DBP by 12.6 and 8.1 mmHg, respectively, and resulting a CBP mean of 134.0/84.5 mmHg at the end of intervention on the SAH stage I patients (CBP mean of 148.5/95.7 mmHg). There were no significant statistical differences between EA and HCTZ interventions on blood pressure decrease, and before-after treatments regarding to biochemical tests and signs of acute toxicity, renal, hepatic and hematologic alterations. A slight trend but no significant difference were observed between adverse events from EA (3.58%) and HCTZ (4.68%) groups. CONCLUSION: EA standardized dry extract was successfully applied to the SAH stage I patient treatment, decreasing effectively SBP ad DBP values to the reference normal ranges, and demonstrating a well-tolerability profile similar to HCTZ intervention.

Chemical composition and chemotaxonomy of Nasutitermes spp. defensive secretion from Brazilian Cerrado for the differentiation of species.

Nasutitermes spp. soldier defensive secretion has a toxic and repellent effect against predators. Chemical profile characterization of this secretion is an interesting tool to differentiate similar termite species. This study aimed to determine defensive secretion composition of Nasutitermes spp. soldier and to apply chemotaxonomy tool for the unambiguous species identification. Fifteen volatile compounds were identified by gas chromatography-mass spectrometry. Multivariate analysis classified populations into three groups. Principal component (PCA), axis 1, was able to separate two groups: group I, colonies 1 and 2 (Nasutitermes corniger) and group II, colony 3 (Nasutitermes ephratae). Therefore, determination of defensive chemical secretion profile proved to be very useful in termite chemotaxonomy, since it was able to differentiate morphologically similar specimens.

Acute toxicity, antinociceptive, and anti-inflammatory activities of the orally administered crotamine in mice.

Crotamine is a polypeptide toxin isolated from rattlesnake venom. Although several studies have been developed identifying many biological effects of isolated crotamine, none of them evaluated its acute toxicity, antinociceptive, and anti-inflammatory activities through oral administration. All in vivo experiments from this study were performed in mice. The up-and-down procedure and hippocratic screening were carried out to evaluate possible pharmacological and toxic effects. Antinociceptive and anti-inflammatory activities of this toxin were evaluated using acetic acid-induced abdominal writhing, formalin-induced pain assays, croton oil-induced ear edema, and carrageenan-induced pleurisy. Crotamine did not cause lethality or signs of intoxication up to the maximum dose tested (10.88 mg/kg). The number of contortions was reduced significantly by 34, 57, and 74% at the oral doses of 0.08, 0.16, and 0.32 mg/kg, respectively. At the dose of 0.16 mg/kg, crotamine decreases pain time-reactivity at neurogenic phase by 45% and at inflammatory phase by 60%. Also, crotamine elicited antiedematogenic activity through the attenuation of the croton oil-induced ear edema by 77%. In the carrageenan-induced pleurisy, the leukocyte, neutrophil, and mononuclear cell migration to the lesion site were reduced by 52%, 46%, and 59%, respectively. Altogether, crotamine demonstrated in vivo antinociceptive and anti-inflammatory effect through acute oral administration, generating an anti-migratory mechanism of action at non-toxic doses.

Food-Effect on (-) - Hydroxycitric Acid Absorption After Oral Administration of Garcinia cambogia Extract Formulation: a Phase I, Randomized, Cross-Over Study.

Garcinia cambogia extract (GCE)/(-) - hydroxycitric acid (HCA) has been identified as a potential antiobesity agent. However, controversial clinical trial results have been published on its biological effect and poor pharmacokinetic (PK) information increases its dubious efficacy. The aim of this study was to determine the main PK parameters of GCE/HCA in healthy women, and to evaluate food effects on HCA absorption. Healthy women ages 21-41 years with body mass index (BMI; kg/m2) 20.29-25.82 participated in a phase I, open-label, randomized, single-dose, cross-over study. In the fasted- and fed-conditions subjects received 1500/750 mg of GCE/HCA under 8 h of fasting. In the fed-period was given a high calorie breakfast (~600 calories) after dosing. Plasma HCA concentrations were substantially reduced in fed-state. Peak plasma concentration (Cmax) and area under the curve of time-concentration (AUC0-10h) were 3-fold and 2-fold lower (p < 0.001, 0.01) in fed-condition, respectively. Higher volume of distribution (Vd/F) and clearance (Cl/F) were achieved in fed state, probably due to the lower fraction (F) of HCA absorbed induced by food effect. Large inter-individual variations were observed for the main pharmacokinetics parameters in both periods. These findings suggest that HCA might suffer an active absorption uptake and intense adsorption on food.

Occupational exposure evaluation of Brazil university community to the volatile organic compounds.

Occupational exposure to volatile organic compounds (VOC) might generate serious worker health damages. Therefore, biological monitoring is essential to evaluate exposure biomarkers from highly toxic chemicals, ensuring better attention to the worker health. In this study was developed and validated a bioanalytical method based on high-performance liquid chromatography coupled to photodiode array (HPLC-PDA) for the quantification of VOC biomarkers in urine samples from Federal University of Goias (UFG) workers. Samples were collected from 30 occupationally exposed subjects after application of a questionnaire survey. The following biomarkers hippuric acid, methyl-hippuric acid, mandelic acid, phenylglyoxylic acid and phenol were quantified, representing exposition to toluene, xylene, styrene, ethylbenzene, benzene and phenol solvents, respectively. Hippuric acid levels were found close to or above the reference values, although a subject had levels higher than preconized by Biological Limit Values (BLV) guideline of 4.0 mg/g creatinine. Five subjects had 3 and 4-methylhippuric acid ranging from 0.1 to 1.0 mg/g creatinine. These results indicate a moderate to high VOC exposure from UFG workers. Multivariate analysis generated four clusters and indicated that histotechnicians and graphic workers need especial attention on occupational VOC exposure. The results from this study reinforce the need for reliable methods able to the biological monitoring as an important tool for assessing occupational exposure.

New bioanalytical method for the quantification of (-) - hydroxycitric acid in human plasma using UPLC-MS/MS and its application in a Garcinia cambogia pharmacokinetic study.

A new, rapid, selective and sensitive UPLC-MS/MS method was developed and validated for the quantification of (-) - hydroxycitric acid (HCA) in human plasma, using DL-malic acid-2,3,3-d3 as internal standard (IS) and simple protein precipitation for the sample preparations. HCA is a highly polar compound make challenging its determination in biological fluids. A specific chromatography column Acquity UPLC HSS T3 (100 × 2.1 mm, 1.8 µm), eluted with mobile phase composed of acetonitrile/ammonium hydroxide 0,1 % (15:85, v/v) were applied for the HCA quantification. The bioanalytical method showed high-throughput achieving as fast chromatographic run as 1 min per sample. No matrix effect was observed with excellent mean chromatographic peak areas ratio of 0.98 ± 0.07 and CV% of 7.17 from normal, lipemic and hemolyzed plasma lots. Calibration curves range was linear at 0.05-10 µg/mL, presenting adequate mean correlation coefficient great than 0.99. Excellent intra-assay and inter-assay precision were achieved, ranging from 5.02-12.01 % (CV%) as well as great intra- and inter-assay accuracy from 0.29-9.20 % (RE%). UPLC-MS/MS bioanalytical method was efficiently applied to the HCA pharmacokinetic study analyzing more than 670 plasma samples.

Aqueous humor concentrations of topical fluoroquinolones alone or in combination with a steroid.

PURPOSE: To compare the aqueous humor (AH) concentrations of moxifloxacin 0.5% and gatifloxacin 0.3% solutions alone or when treatment was combined with steroids, and to correlate these concentrations with the minimum inhibitory concentrations (MIC) for the most common endophthalmitis-causing organisms. METHODS: Patients undergoing phacoemulsification were enrolled to receive one drop of one of the following solutions: moxifloxacin (G1), moxifloxacin + dexamethasone (G2), gatifloxacin (G3), or gatifloxacin + c (G4), every 15 min, 1h before surgery. AH samples were collected before surgery and analyzed using HPLC-tandem mass spectrometry. RESULTS: The mean antibiotic concentrations in the AH were: G1= 1280.8 ng/mL; G2= 1644.3 ng/mL; G3= 433.7 ng/mL; and G4= 308.1 ng/mL. The mean concentrations statistically differed between G1 and G2 (p=0.01), and G3 and G4 (p=0.008). All samples achieved the MIC for Staphylococcus epidermidis; 100% of the samples from G1 and G2, and 97% from G3 and G4 reached the MIC for fluoroquinolone-sensitive Staphylococcus aureus; 100% of the samples from G1 and G2, 88% from G3, and 72% from G4 reached the MIC for enterococci (p<0.001); and 100% of samples from G1 and G2, 59% from G3, and 36% from G4 reached the MIC for Streptococcus pneumoniae (p<0.001). For fluoroquinolone-resistant S. aureus, 23% from G1, 44% from G2, and no samples from G3 or G4 achieved the MIC (p<0.001). CONCLUSIONS: Moxifloxacin + dexamethasone demonstrated a higher concen-tration in the AH than the moxifloxacin alone. Gatifloxacin + steroids demonstrated less penetration into the anterior chamber than gatifloxacin alone. Moxifloxacin was superior to gatifloxacin considering the MIC for enterococci, S. pneumoniae, and fluoroquinolone-resistant S. aureus.

Aqueous humor concentrations of topical fluoroquinolones alone or in combination with a steroid / Concentração de fluorquinolonas no humor aquoso após instilação tópica das associações com corticosteroides

ABSTRACT Purpose: To compare the aqueous humor (AH) concentrations of moxifloxacin 0.5% and gatifloxacin 0.3% solutions alone or when treatment was combined with steroids, and to correlate these concentrations with the minimum inhibitory concentrations (MIC) for the most common endophthalmitis-causing organisms. Methods: Patients undergoing phacoemulsification were enrolled to receive one drop of one of the following solutions: moxifloxacin (G1), moxifloxacin + dexamethasone (G2), gatifloxacin (G3), or gatifloxacin + c (G4), every 15 min, 1h before surgery. AH samples were collected before surgery and analyzed using HPLC-tandem mass spectrometry. Results: The mean antibiotic concentrations in the AH were: G1= 1280.8 ng/mL; G2= 1644.3 ng/mL; G3= 433.7 ng/mL; and G4= 308.1 ng/mL. The mean concentrations statistically differed between G1 and G2 (p=0.01), and G3 and G4 (p=0.008). All samples achieved the MIC for Staphylococcus epidermidis; 100% of the samples from G1 and G2, and 97% from G3 and G4 reached the MIC for fluoroquinolone-sensitive Staphylococcus aureus; 100% of the samples from G1 and G2, 88% from G3, and 72% from G4 reached the MIC for enterococci (p<0.001); and 100% of samples from G1 and G2, 59% from G3, and 36% from G4 reached the MIC for Streptococcus pneumoniae (p<0.001). For fluoroquinolone-resistant S. aureus, 23% from G1, 44% from G2, and no samples from G3 or G4 achieved the MIC (p<0.001). Conclusions: Moxifloxacin + dexamethasone demonstrated a higher concen­tration in the AH than the moxifloxacin alone. Gatifloxacin + steroids demonstrated less penetration into the anterior chamber than gatifloxacin alone. Moxifloxacin was superior to gatifloxacin considering the MIC for enterococci, S. pneumoniae, and fluoroquinolone-resistant S. aureus.

RESUMO Objetivos: Comparar a concentração no humor aquoso entre as soluções de moxifloxacina 0,5% e gatifloxacina 0,3% sozinhas ou combinadas com corticosteroides, e correlacionar a concentração com a concentração inibitória mínima (MIC) para os agentes microbianos mais comumente relacionados a endoftalmites. Métodos: Pacientes que seriam submetidos a cirurgia de catarata foram selecionados para receber 1 gota a cada 15 min, 1 hora antes do procedimento de uma das seguintes soluções: moxifloxacina (G1), moxifloxacina + dexametasona (G2), gatifloxacina (G3) ou gatifloxacina + prednisolona (G4). Amostras do humor aquoso foram coletadas antes do início da cirurgia. Espectrofotometria de massa por HPLC determinou a concentração do antibiótico nas amostras. Resultados: A concentração media de antibiótico nas amostras foram: G1= 1280,8 ng/mL; G2= 1644,3 ng/mL; G3= 433,7 ng/mL; G4= 308,1 ng/mL. Concentração média entre G1 e 2 (p=0,01), e G3 e 4 (p=0,008). Todas as amostras alcançaram MIC para S. epidermidis; 100% das amostras do G1 e 2, e 97% do G3 e 4 atingiram o MIC para S. aureus fluoroquinolona-sensível; 100% das amostras do G1 e 2, 88% do G3 e 72% do G4 atingiram o MIC para Enterococci (p<0,001); e 100% das amostras do G1 e 2, 59% do G3 e 36% do G4 atingiram o MIC para S. pneumoniae (p<0,001). Para o S. aureus resistente a fluoroquinolona, 23% do G1, 44% do G2, e nenhuma das amostras G3 e 4 atingiram o MIC (p<0,001). Conclusão: Moxifloxacina + dexamethasona demonstrou maior concentração no humor aquoso comparado com a moxifloxacina sozinha. Gatifloxacina + esteróide demonstrou menor penetração na câmara anterior comparado a solução de ga­ti­floxacin sem corticóide. A moxifloxacina foi superior a gatifloxacina considerando o MIC para Enterococci, S. pneumoniae e S. aureus fluorquinolona resistente.

Aqueous Humor Penetration and Biological Activity of Moxifloxacin 0.5% Ophthalmic Solution Alone or with Dexamethasone 0.1.

PURPOSE: To compare aqueous humor concentrations of topically applied moxifloxacin 0.5% ophthalmic solution alone or in combination with dexamethasone 0.1% and to correlate these concentrations with the minimum inhibitory concentrations (MICs) for common endophthalmitis-causing organisms. METHODS: Sixty-eight patients undergoing routine phacoemulsification with intraocular lens implantation received either moxifloxacin 0.5% alone or moxifloxacin 0.5% combined with dexamethasone. For both groups, 1 drop of the test solution was instilled 4 times daily 1 day preoperatively and 1 drop 1 h preoperatively. An aqueous humor sample obtained immediately before paracentesis was submitted to high-performance liquid chromatography-tandem mass spectrometry to determine the moxifloxacin concentration. RESULTS: The mean concentrations of moxifloxacin were 986.6 ng/mL in the moxifloxacin with dexamethasone group and 741.3 ng/mL in the moxifloxacin group (P = 0.13). Moxifloxacin concentrations of all samples exceeded the MICs for Staphylococcus epidermidis, S. aureus, and Streptococcus pneumoniae. All samples in the moxifloxacin with dexamethasone group and 94% in the moxifloxacin group achieved the MIC for Enterococcus species. For quinolone-resistant S. aureus, the MIC was achieved in 29% in the moxifloxacin with dexamethasone group and 9% in the moxifloxacin group (P = 0.06). CONCLUSION: Aqueous humor moxifloxacin concentrations were higher when topically administrated in combination with dexamethasone compared to the moxifloxacin alone. However, this difference was not statistically significant. Nevertheless, the MICs of the most common pathogens associated with endophthalmitis were exceeded in both study groups.

Automated determination of venlafaxine in human plasma by on-line SPE-LC-MS/MS. Application to a bioequivalence study.

A new automated SPE-LC-ESI-MS/MS method was developed and validated to quantify venlafaxine in human plasma using fluoxetine as an internal standard. The analytes were automatically extracted from plasma by C18 SPE cartridges, separated on a C8 RP column and analyzed by MS in the multiple reaction-monitoring (MRM) mode. The method has a chromatographic run time of 4.0 min and a linear calibration curve over the range of 0.25-200 ng/mL (r >0.997). The between-run precisions, based on the percent RSD for replicate quality controls (0.75; 80, and 200 ng/mL), were < 8.5% for all concentrations. The between-run accuracies, based on the percent relative error, were < 4.0%. This method was successfully employed in a bioequivalence study of two venlafaxine capsule formulations (test formulation from Eurofarma (Brazil) and Efexor XR, reference formulation, from Wyeth-Whitehall, Brazil) in 48 healthy volunteers of both sexes who received a single 150 mg dose of each formulation. More than 3000 samples were analyzed eliminating the analyst's exposure to hazardous organic solvents normally employed in off-line liquid-liquid extractions. The 90% confidence interval (CI) of the individual ratio geometric mean for Test/Reference was 91.6-103.4% for AUC(0-48 h) and 102.2-112.6% for C(max). Since both 90% CI for AUC(0-48 h) and C(max) were included in the 80-125% interval proposed by the US Food and Drug Administration (FDA) and the Brazilian National Health Surveillance Agency (ANVISA), the test formulation was considered bioequivalent to Efexor XR according to both the rate and extent of absorption.