Synthesis and antibacterial activity of water-soluble limonin benzoylhydrazone derivatives / 中国药科大学学报

@#As a compound rich in citrus plants, limonin has a wide range of biological activities, but its practical application is limited because of its poor water solubility.In this paper, eight new compounds 7a-7h were designed and synthesized by introducing benzoyl hydrazone at the carbonyl position of limonin.The results showed that the water solubilities of all compounds were higher than that of limonin, especially 7a, 7d, 7e and 7f.In addition, the experiment showed that compounds 7d and 7e with substituted hydroxyl groups at the interposition and para-position of the benzene ring had strong antibacterial effects against Escherichia coli and Staphylococcus aureus, and that compound 7e had the best activity, with minimum inhibitory concentrations of 0.31 mg/mL and 1.25 mg/mL, respectively.And compound 7e had better antibacterial activiy in E.coli than sodium benzoate.

Synthesis and antileishmanial activity of naphthoquinone-based hybrids / Síntesis y actividad antileishmania de híbridos naftoquinónicos / Síntese e atividade antileishmania de híbridos naftoquinónicos

SUMMARY Introduction: Leishmaniasis is a disease caused by protozoa of the genus Leishmania and is considered endemic in 98 countries. Treatment with pentavalent antimonials has a high toxicity, which motivates the search for effective and less toxic drugs. α- and β-lapachones have shown different biological activities, including antiprotozoa. In recent studies, the isonicotinoylhydrazone and phthalazinylhydrazone groups were considered innovative in the development of antileishmania drugs. Molecular hybridization is a strategy for the rational development of new prototypes, where the main compound is produced through the appropriate binding of pharmacophoric subunits. Aims: To synthesize four hybrids of α- and β-lapachones, together with the isonicotinoylhydrazone and phthalazinylhydrazone groups and to determine the antileishmania activity against the promastigotic forms of L. amazonensis, L. infantum and L. major. Results: β-lapachone derivatives were more active against all tested leishmania species. βACIL (IC50 0.044μM) and βHDZ (IC50 0.023μM) showed 15-fold higher activity than amphotericin B. The high selectivity index exhibited by the compounds indicates greater safety for vertebrate host cells. Conclusion: The results of this work show that the hybrids βACIL and (3HDZ are promising molecules for the development of new antileishmania drugs.

RESUMEN Introducción: Leishmaniasis es una enfermedad causada por protozoos del género Leishmania y se considera endémica en 98 países. El tratamiento con antimoniales pentavalentes tiene una alta toxicidad, lo que motiva la búsqueda de fármacos eficaces y menos tóxico. α- y β-lapachones han mostrado diferentes actividades biológicas, incluido los antiprotozoarios. En estudios recientes, los grupos isonicotinoilhidra-zona y ftalazinilhidrazona se consideraron innovadores en el desarrollo de fármacos antileishmania. La hibridación molecular es una estrategia para el desarrollo racional de nuevos prototipos, donde el compuesto principal se produce a través de la unión apropiada de subunidades farmacofóricas. Objetivos: Sintetizar cuatro híbridos de α- y β-lapachones, junto con los grupos isonicotinoilhidrazona y ftalazinilhidrazona y determinar la actividad antileishmania frente a las formas promastigotas de L. amazonensis, L. infantum y L. major. Resultados: Los derivados de β-lapachone fueron más activos contra todas las especies de leishmania probadas. La βACIL (CI50 0,044μM) y βHDZ (CI50 0,023μM) mostraron actividad 15 veces mayor que la anfotericina B. El alto índice de selectividad que presentan los compuestos indica una mayor seguridad para las células huésped del vertebrado. Conclusión: Los resultados de este trabajo demuestran que los híbridos (ACIL y (HDZ son moléculas prometodoras para el desarrollo de nuevos fármacos antileishmania.

RESUMO Introdução: A leishmaniose é uma doença causada por protozoários do género Leishmania e é considerada endémica em 98 países. O tratamento com antimoniais pentavalentes apresenta alta toxicidade, o que motiva a pesquisa por medicamentos eficazes e menos tóxicos. α- e β-lapachones tém mostrado diferentes atividades biológicas, incluindo antiprotozoários. Em estudos recentes, os grupos isonicotinoilhi-drazona e ftalazinilhidrazona foram considerados inovadores no desenvolvimento de drogas antileishmania. A hibridização molecular é uma estratégia para o desenvolvimento racional de novos protótipos, onde o composto principal é produzido através da ligação apropriada de subunidades farmacofóricas. Objetivos: Sintetizar quatro híbridos de α- e β-lapachones, juntamente com os grupos isonicotinoil-hidra-zona e ftalazinilhidrazona e determinar a atividade antileishmania contra as formas promastigóticas de L. amazonensis, L. infantum e L. major. Resultados: Os derivados de β-lapachona foram mais ativos contra todas as espécies de leishmania testadas. BACIL (IC50 0,044 μM) e βHDZ (IC50 0,023 μM) apresentaram atividade 15 vezes maior do que a anfotericina B. O alto índice de seletividade dos compostos indica maior segurança para células hospedeiras de vertebrados. Conclusaõ: Os resultados deste trabalho mostram que os híbridos βACIL e βHDZ são moléculas promissoras para o desenvolvimento de novos fármacos antileishmania.

Structural, spectroscopic, and theoretical analysis of a molecular system based on 2-((2-(4-chlorophenylhydrazone) methyl)quinolone / Análisis estructural, espectroscópico y teórico de un sistema molecular basado en la 2-((2-(4-clorofenilhidrazono) metil)quinolina / Análise estrutural, espectroscópico, e teórico de um sistema molecular com base na 2-((2-(4-Clorofenilhidrazona) metil)quinolina

Abstract A novel molecular system based on 2-((2-(4-chlorophenylhydrazone)methyl)quinoline (1-E ) was synthesized. Interconversion of 1-E to its configurational isomer 1-Z was achieved using UV radiation (250 W Hg lamp). Such isomerization was monitored by ¹H-NMR. The results suggest that the hydrazone derivative can act as a chemical brake in solution. This molecular system was structurally (Single Crystal X-Ray diffraction and DFT calculations) and spectroscopically (NMR, UV, and IR) characterized. Electrochemical measurements showed that configurational changes induce differential redox behavior. In this regard, the reported quinoline system exhibits different dynamic absorption and electrochemical properties that are modulated by UV-light. Therefore, 1-E can be regarded as a potential photo-electrochemical switch.

Resumen Se sintetizó un nuevo sistema molecular basado en 2-((2-(4-chlorofenilhidrazona)metil)quinolina. Del mismo modo, se evaluó la respuesta dinámica de este compuesto a radiación ultravioleta y formación de un enlace de hidrógeno intramolecular. Los resultados muestran que este derivado de hidrazona puede actuar como freno en solución. El sistema en mención es descrito estructural (Cristalografía de Rayos X y cálculos DFT) y espectroscópicamente (RMN, UV e IR). La interconversión de este sistema entre las configuraciones 1-E y 1-Z fue mediada por radiación UV y monitoreada a través de RMN-¹H. El estudio electroquímico mostró un comportamiento diferencial en función de su configuración, aspecto fundamental en el desarrollo de sistemas foto- y electroquímicamente modulados.

Resumo Neste trabalho é apresentado um novo sistema molecular baseado na 2-((2-(4-clorofenilhidrazona)metil)quinolina, capaz de responder dinamicamente à radiação ultravioleta formando uma ligação de hidrogénio intramolecular que atua como um freio na solução. Este sistema é descrito estruturalmente (cristalografia de raios-X e DFT) e por diferentes técnicas espectroscópicas (RMN, de UV e de IV). Radiação UV foi usada para fazer a interconversão da hidrazona 1-E no seu isômero configuracional 1-Z . Este processo foi monitorado pelo RMN. As medidas eletroquímicas mostraram que as mudanças configuracionais entre os isômeros induzem a comportamentos redox diferentes, o que é uma caraterística chave no desenvolvimento de interruptores fotoelectroquímicos.

Discovery of N-arylsulfonyl-3-acylindole benzoyl hydrazone derivatives as anti-HIV-1 agents

The discovery and development of novel inhibitors with activity against variants of human immunodeficiency virus type 1 (HIV-1) is pivotal for overcoming treatment failure. As our ongoing work on research of anti-HIV-1 inhibitors, 32 N-arylsulfonyl-3-acylindole benzoyl hydrazone derivatives were prepared by introduction of the hydrazone fragments on the N-arylsulfonyl-3-acylindolyl skeleton and preliminarily screened in vitro as HIV-1 inhibitors for the first time. Among of all the reported analogues, eight compounds exhibited significant anti-HIV-1 activity, especially N-(3-nitro)phenylsulfonyl-3-acetylindole benzoyl hydrazone (18) and N-(3-nitro)phenylsulfonyl-3-acetyl-6-methylindole benzoyl hydrazone (23) displayed the most potent anti-HIV-1 activity with EC50 values of 0.26 and 0.31 µg/mL, and TI values of >769.23 and >645.16, respectively. It is noteworthy that introduction of R3 as the methyl group and R2 as the hydrogen group could result in more potent compounds. This suggested that introduction of R3 as the methyl group could be taken into account for further preparation of these kinds of compounds as anti-HIV-1 agents

Tat-functionalized Ag-FeO nano-composites as tissue-penetrating vehicles for tumor magnetic targeting and drug delivery

In this paper, we prepared a dual functional system based on dextrin-coated silver nanoparticles which were further attached with iron oxide nanoparticles and cell penetrating peptide (Tat), producing Tat-modified Ag-FeO nanocomposites (Tat-FeAgNPs). To load drugs, an -SH containing linker, 3-mercaptopropanohydrazide, was designed and synthesized. It enabled the silver carriers to load and release doxorubicin (Dox) in a pH-sensitive pattern. The delivery efficiency of this system was assessed using MCF-7 cells, and using null BalB/c mice bearing MCF-7 xenograft tumors. Our results demonstrated that both Tat and externally applied magnetic field could promote cellular uptake and consequently the cytotoxicity of doxorubicin-loaded nanoparticles, with the IC of Tat-FeAgNP-Dox to be 0.63 µmol/L. The delivery efficiency of Tat-FeAgNP carrying Cy5 to the mouse tumor was analyzed using the optical imaging tests, in which Tat-FeAgNP-Cy5 yielded the most efficient accumulation in the tumor (6.7±2.4% ID of Tat-FeAgNPs). Anti-tumor assessment also demonstrated that Tat-FeAgNP-Dox displayed the most significant tumor-inhibiting effects and reduced the specific growth rate of tumor by 29.6% ( = 0.009), which could be attributed to its superior performance in tumor drug delivery in comparison with the control nanovehicles.

Synthesis and Characterization of Novel Hydrazone Based Anti-mutagenic and Antioxidative Agents.

The present study was designed to produce novel hydrazine and evaluate their biological properties including antioxidant, antityrosinase and antimutagenic. 4-allyloxybenzoyl hydrazine (1) reacts with 5-acetyl-1,3-dimethyl barbituric acid (2) and 2-Isonitrosoacetophenone (3) in the presence of acetic acid as a catalyst to produce the hydrazone derivatives 4 and 5 in high yields respectively. The new hydrazone derivatives 4 and 5 have been fully characterized by using multinuclear NMR (1H, 13C) spectroscopy and elemental analysis. The compounds 4 and 5 were studied for their antioxidant and tyrosinase enzyme inhibition activity. In addition the mutagenic and antimutagenic activities were evaluated by Ames Salmonella/ microsome mutagenicity test. The results showed that both of compounds exhibited significant antioxidative and antimutagenic activity and compound 5 has shown moderate tyrosinase inhibition activity. This study suggested that these compounds could be considered as novel bioactive agents in pharmaceutical area.

Title: Development of new spectrophotometric method for estimation of tenofovir disoproxil fumarate using MBTH reagent.

A new simple, sensitive and specific procedure has been developed for determination of tenofovir disoproxil fumarate in bulk and pharmaceutical dosage forms using MBTH reagent. The purpose of this analytical validation procedure is to validate it by laboratory experiments to prove that the method meets the minimum standards for laboratory use. 3-methyl-2-bezothiazoline hydrazone reacts with the secondary amine group of tenofovir in the presence of oxidizing agent, ferric chloride. The resulting apple green coloured chromogen when measured spectrophotometrically in visible region (i.e., 400-800nm) shows a maximum absorbance at 626.5nm. This method can be successfully applied for the determination of drug content in pharmaceutical formulations. The results of analysis have been validated statistically.

Synthesis and antitumor activity of fluoroquinolon-3-yl s-triazole sulfanylacetylhydrazones and s-triazole hydrazone derivatives (V) / 中国药学杂志

OBJECTIVE: To explore an efficient modification strategy for conversion of antibacterial fluoroquinolones to antitumor ones. METHODS: An azole heterocyclic ring, s-triazole, as a bioisotere of the C-3 carboxylic acid for ofloxacin(1), functional acyl-hydrazones and hydrazones, was used as the modified side-chain for the C-3 bioisotere, then the C-3 s-triazole acylhydrazones and s-triazole hydrazone derivatives were designed and their in vitro antitumor activity was evaluated by MTT assay. RESULTS: Fourteen target compounds were synthesized, and they exhibited stronger antitumor activity than the parent ofloxacin. And most importantly, hydrazone derivatives had higher activity than their corresponding acylhydrazone compounds. CONCLUSION: s-Triazole-hydrazone moiety is warranted special attention as an efficient bioisosteric replacement of the C-3 carboxylic acid for further development of antitumor fluoroquinolone lead compounds.

Spread of Efflux Pump Overexpressing-Mediated Fluoroquinolone Resistance and Multidrug Resistance in Pseudomonas aeruginosa by using an Efflux Pump Inhibitor / 감염과화학요법

BACKGROUND: Fluoroquinolone resistance in Pseudomonas aeruginosa may be due to efflux pump overexpression and/or target mutations. We designed this study to investigate the efflux pump mediated fluoroquinolone resistance and check the increasing effectiveness of fluoroquinolones in combination with an efflux pumps inhibitor among P. aeruginosa isolates from burn wounds infections. MATERIALS AND METHODS: A total of 154 consecutive strains of P. aeruginosa were recovered from separate patients hospitalized in a burn hospital, Tehran, Iran. The isolates first were studied by disk diffusion antibiogram for 11 antibiotics and then minimum inhibitory concentration (MIC) experiments were performed to detect synergy between ciprofloxacin and the efflux pump inhibitor, carbonyl cyanide-m-chlorophenyl hydrazone (CCCP). Then to elucidate the inducing of multi drug resistance due to different efflux pumps activation in Fluoroquinolone resistant isolates, synergy experiments were also performed in random ciprofloxacin resistant isolates which have overexpressed efflux pumps phenotypically, using CCCP and selected antibiotics as markers for Beta-lactams and Aminoglycosides. The isolates were also tested by polymerase chain reaction (PCR) for the presence of the MexA, MexC and MexE, which encode the efflux pumps MexAB-OprM, MexCD-OprJ and MexEF-OprN. RESULTS: Most of the isolates were resistant to 3 or more antibiotics tested. More than half of the ciprofloxacin resistant isolates exhibited synergy between ciprofloxacin and CCCP, indicating the efflux pump activity contributed to the ciprofloxacin resistance. Also increased susceptibility of random ciprofloxacin resistant isolates of P. aeruginosa to other selected antibiotics, in presence of CCCP, implied multidrug extrusion by different active efflux pump in fluoroquinolones resistant strains. All of Ciprofloxacin resistant isolates were positive for MexA, MexC and MexE genes simultaneously. CONCLUSION: In this burn hospital, where multidrug resistant P. aeruginosa isolates were prevalent, ciprofloxacin resistance and multidrug resistance due to the overexpression of fluoroquinolones mediated efflux pumps has also now emerged. Early recognition of this resistance mechanism should allow the use of alternative antibiotics and use an efflux pumps inhibitor in combination with antibiotic therapy.

Development of new spectrophotometric method for estimation of tenofovir disoproxil fumarate using MBTH reagent.

A new simple, sensitive and specific procedure has been developed for determination of tenofovir disoproxil fumarate in bulk and pharmaceutical dosage forms using MBTH reagent. The purpose of this analytical validation procedure is to validate it by laboratory experiments to prove that the method meets the minimum standards for laboratory use. 3-methyl-2-bezothiazoline hydrazone reacts with the secondary amine group of tenofovir in the presence of oxidizing agent, ferric chloride. The resulting apple green coloured chromogen when measured spectrophotometrically in visible region (i.e., 400-800nm) shows a maximum absorbance at 626.5nm. This method can be successfully applied for the determination of drug content in pharmaceutical formulations. The results of analysis have been validated statistically.

Design, synthesis and spectral characterization of 5-[2(3)-dialkylamino alkoxy] indole 3-hydrazone 2-one and 5-hydroxyindole 3-thiosemicarbazone 2-ones.

In the present work, some new 5-[2(3)-dialkylamino alkoxy] Indole 3-hydrazone 2-one and 5-[2(3)-dialkylamino alkoxy] indole 3-thiosemicarbazone 2- ones were prepared from 5-hydroxy isatin. The structures of the products were characterized by IR, NMR, MASS Spectral studies. Simplicity of the reaction conditions, easy workup procedure and good yields are the key features of this protocol.

Synthesis and antitumor activity fluoroquinolon-3-yloxadiazole sulfanylacetylhydrazone derivatives / 中国药学杂志

OBJECTIVE: To explore an efficient modification strategy for a bioisotere antibacterial fluoroquinolon-3-yl carboxylic acid group for developing compounds with improved antitumor activity. METHODS: Novel title 2-(fluoroquinolon-3-yl)-oxadiazole-5-sulfanylacetylhydrazone derivatives were designed on the basis the pharmacophore skeleton transition principle, and the in vitro activity the title compounds was evaluated by MTT assay. RESULTS: Fourteen target compounds were synthesized from ofloxacin and exhibited more potent antitumor activity than the parent compound, especially the compounds with a carboxylic group-substitued phenyl ring as a modified group around the bioisostere, which had comparable activity to doxorubicin. CONCLUSION: A hydrazone with electron-withdrawing substituents warrants further development as the modified side-chain for the C-3 bioisostere.

Relationship between efflux pump phenotype and genotype in multiple drug Acinetobacter baumannii / 中华检验医学杂志

Objective To expore multi-drug resistant Acinetobacter baumannii efflux pump phenotype and efflux pump gene expression in the resistant isolates. Methods Application of K-B method to detect 96 strains isolated from the First Hospital of Hebei Medical University multi-drug resistant Acinetobacter baumannii′ resistance to 15 kinds of antibacterial drugs, detecting multi-drug resistant Acinetobacter baumannii efflux pump phenotype with broth microdilution method by the addition of carbonyl cyanide chlorobenzene hydrazone ( CCCP) pump inhibitors,using PCR amplification and sequencing to study efflux pump protein gene sequence characteristics . Results The Acinetobacter baumannii resistance rate of 96 strains to quinolones, cephalosporins, aminoglycosides, tetracyclines were 70. 8%-94. 8%.There were 34 positive efflux pump phenotypes in 96 multi-drug resistant Acinetobacter baumannii strains, including 33 adeB strains, 32 adeR strains, 33 adeS strains, 33 adeJ strains,0 adeE strain,33 adeM strains, positive detection rate were 97. 06%, 94. 12%, 97. 06%, 97. 06%, 0, 97. 06%, respectively. By sequence comparison, adeB, adeR and adeS genes sequence homology was 100% in the GenBank. Conclusion Active efflux pump gene perturbation is one of the important factors in multi-drug resistant Acinetobacter baumannii.

Establishment of quantitative determination methods of total glycoside groups composition in Chinese materia medica by differential anthrone-sugar hydrazone / 中草药

Objective: To establish a method for the determination of total glycosides in Chinese materia medica (CMM)-differential anthrone-sugar hydrazone colorimetry. Methods: The content of total glycosides (including sugar) in CMM was determined by anthrone colorimetry, and then sugar hydrazone-sulfuric acid colorimetric method was used to determine the content of reducing sugars. The content of total glycosides came from the two subtraction. Results: Anthrone colorimetric reaction was completed within 6 h, detection wavelength was 620 nm, linear range was 80-180 μg/mL, correlation coefficient R2 = 0.994, and recovery was 99.2%, RSD was 1.35%. Sugar hydrazone-sulfuric acid color reaction method was completed within 4 h, detection wavelength was 412 nm, linear range was 2-12 mg/mL, correlation coefficient R2 = 0.991, and recovery was 99.5%, RSD was 1.98%. The total glycosides contents in five Chinese patent medicines such as Buyang Huanwu Decoction, Liuwei Dihuang Pill, Andrographis Tablet, Astragalus Injection, and Pachymaran Oral Liquid were 15.1, 26.0, 6.68 mg/g, and 21.3, 16.9 mg/mL. Conclusion: This method is firstly established for the quantitative determination of total glycosides in CMM, and the ingredient group analysis method is established by anthrone colorimetry and sugar hydrazone-sulfuric acid colorimetric method. The methods have better accuracy, and can be used for the determination of total glycosides ingredients group of CMM.

Influence of cyanide p-trifluoromethoxyphenyl-hydrazone on insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes / 中华实用儿科临床杂志

Objective To observe the effect of uncoupler of mitochondrial oxidative phosphorylation——Cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP) on mitochondrial function and insulin sensitivity in mature adipocytes.Me-thods 3T3-L1 pre-adipocytes were differentiated into mature adipocytes and then induced and maintained in medium that contained the chemical uncoupler 7.5 μmol/L FCCP.Glucose uptake was determined in the adipocytes by measu-ring 2-deoxy-D-[3H] glucose uptake.Western blot was used to detect the translocation of insulin-sensitive glucose transporter 4 (GLUT4) and measure the phosphorylation and total protein contents of insulin signaling proteins such as insulin receptor substrate(IRS)-1,Akt.The mitochondrial morphology was performed by transmission electron microscope.The mitochondrial DNA (mtDNA) copy number was evaluated by real time PCR.Luciferase-based luminescence assay was used to determine cellular ATP production.The mitochondrial membrane potential(△Ψm) and reactive oxygen species(ROS) were detected by flow cytometry.Results (1) Exposure of mature adipocytes to FCCP basal glucose uptake was similar to mature adipocytes without FCCP(t =-0.07,P ＞ 0.05) ; however,the insulin-stimulated glucose uptake was significantly decreased in FCCP group (t =5.87,P ＜ 0.01).(2)FCCP decreased insulin-stimulated GLUT4 translocation to the plasmalemma and inhibition of insulin-induced phosphorylation of IRS-1 and Akt in 3T3-L1 adipocytes.(3)The size of mitochondria in FCCP-treated adipocytes was smaller than that in 3T3-L1 adipocytes without FCCP,and the morphology was condensed and abnormal.(4) mtDNA copy number in FCCP-treated adipocytes was significantly lower than that in adipocytes without FCCP(t =-1.73,P ＜ 0.001).(5) Exposure of 3T3-L1 adipocytes to FCCP significantly decreased △Ψm (t =4.83,P ＜ 0.01) and total cellular ATP production compared with cells without FCCP (t =6.08,P ＜ 0.0001),as well as the increased intracellular ROS levels (t =-6.82,P ＜ 0.01).Conclusions FCCP may impair mitochondrial morphology and mitochondrion dysfunction,and inhibited the activation of insulin-induced phosphorylation of IRS-1 and Akt in 3T3-L1 adipocytes,which suggests that FCCP-induced insulin resistance may be correlated with the FCCP induced generation of ROS in 3T3-L1 adipocytes.

Sensitive spectrophotometric determination of thorium (IV) using diacetyl monoxime isonicotinoyl hydrazone (DMIH).

Thorium (IV) forms a yellow Coloured water soluble complex with Diacetyl Monoxime Isonicotinoyl hydrazone (DMIH) reagent in acidic buffer of PH 5.0 with λ max at 352 nm. The molar absorptivity and sandell’s sensitivity are 2.265 X 10 4 L mol -1 cm -1 and 0.0022μg /cm2 respectively. The Beer’s law validity range 1.16–13.92 μg / mL. Thorium (IV) forms (M: L) 1:1 complex with DMIH and stability constant of the complex is 7.0265 X 10 6.The derivative spectrophotometric determination of Th (IV)) was carried out by measuring peak height method. The developed derivative spectrophotometric method was employed for the determination of Thorium (IV) in synthetic samples of alloy. The effect of various diverse ions was also studied.

Anti-parasitic action and elimination of intracellular Toxoplasma gondii in the presence of novel thiosemicarbazone and its 4-thiazolidinone derivatives

Toxoplasma, which infects all eukaryotic cells, is considered to be a good system for the study of drug action and of the behavior of infected host cells. In the present study, we asked if thiosemicarbazone derivatives can be effective against tachyzoites and which morphological and ultrastructural features of host cells and parasites are associated with the destruction of Toxoplasma. The compounds were tested in infected Vero cell culture using concentration screens (0.1 to 20 mM). The final concentration of 1 mM was chosen for biological assay. The following results were obtained: 1) These new derivatives decreased T. gondii infection with an in vitro parasite IC50 percent of 0.2-0.7 mM, without a significant effect on host cells and the more efficient compounds were 2, 3 (thiosemicarbazone derivatives) and 4 (thiazolidinone derivative); 2) The main feature observed during parasite elimination was continuous morphological disorganization of the tachyzoite secretory system, progressive organelle vesiculation, and then complete disruption; 3) Ultrastructural assays also revealed that progressive vesiculation in the cytoplasm of treated parasites did not occur in the host cell; 4) Vesiculation inside the parasite resulted in death, but this feature occurred asynchronously in different intracellular tachyzoites; 5) The death and elimination of T. gondii was associated with features such as apoptosis-like stage, acidification and digestion of parasites into parasitophorous vacuoles. Our results suggest that these new chemical compounds are promising for the elimination of intracellular parasites by mainly affecting tachyzoite development at 1 mM concentration for 24 h of treatment.

Determinação espectrofotométrica do bromidrato de fenoterol em preparações farmacêuticas / Spectrophotometric determination of fenoterol hydrobromide in pharmaceutical preparations

Um método espectrofotométrico simples foi desenvolvido para a determinação do bromidrato de fenoterol (BF) em comprimidos, gotas e xarope, como princípio ativo único e associado aoibuprofeno. O método se baseia na reação de acoplamento oxidativo do BF com o 3-metil-2-benzotiazolinona hidrazona (MBTH), na presença de sulfato cérico, como agente oxidante. Amistura de BF, MBTH e sulfato cérico, em meio ácido, produz um composto colorido (vermelho castanho) com máximo de absorção a 475 nm. A curva de calibração foi linear num intervalo deconcentração de 3,0 a 12,0 μg/mL, com coeficiente de correlação linear de 0,9998. Os parâmetros experimentais que afetam o desenvolvimento e a estabilidade do produto colorido foramcuidadosamente estudados e otimizados. O método foi aplicado em amostras comerciais e simuladas, obtendo-se coeficientes de variação entre 0,25 % a 0,82 % e médias de recuperação do padrão que variaram de 98 % a 102 %. O método proposto mostrou-se exato, preciso, linear e não é passível de interferência de excipientes, para as formas farmacêuticas comprimidos e gotas. Não houve interferência do ibuprofeno que consta de uma dasformulações analisadas, associado ao BF. Quanto ao xarope, houve interferência do veículo sugerindo reações de seus componentes com o MBTH.

A simple spectrophotometric method has been developed for the determination of fenoterol hydrobromide (FH) in TABELA IV - Valores obtidos em termos de fatores de resposta (FR), considerando a concentração de BF e respectivas absorvâncias determinadas em 475 nm, a partir do placebo da amostra B. Os valores expressam a média de três determinações Concentração Concentração Absorvância Fator deteórica encontrada Resposta (μg/mL) (μg/mL) (FR) 6,0 5,93 0,4895 0,082557,0 6,89 0,5633 0,08176 8,0 8,12 0,6571 0,08092 9,0 9,09 0,7471 0,08219 10,0 10,04 0,8144 0,08112FR médio = 0,08171; DP = 0,0006912; CV = 0,85%. tablets, drops and syrup, as the only active principle andassociated with ibuprofen. The method is based on the oxidative coupling reaction of the FH with 3-methyl-2-benzothiazolinone hydrazone (MBTH) and ceric sulphate as oxidant reagent. The mixture of the drug, MBTH andceric sulfate, in acid medium, produces a red brown color compound, with absorption maximum at 475 nm. Thecalibration curve was linear over a concentration range from 3.0 to 12.0 μg/mL, with correlation coefficient of0.9998. The different experimental parameters affecting the development and stability of the color compound werecarefully studied and optimized. The method was applied successfully to assay FH in dosage forms and simulatedsamples. The coefficient of variation was from 0.25 % to 0.82 % and average recoveries of the standard from 98 % to 102 %. The excipients (tablets and drops) did not interfere in the analysis and the results showed that method can be used for determination of the FH isolated or associated with ibuprofen with precision, accuracy and specificity. In case of syrup, the interference in the analysis suggests apossible reaction between vehicle components with MBTH.

The association between resistance phenotypes and expression levels of efflux pumps of Pseudomonas aeruginosa / 中华检验医学杂志

Objective To study the effects of efflux pump inhibitors(CCCP and PAβN)on carbapenems in Pseudomonas aernginosa(P.aeruginosa)clinical isolates and investigate the association between the resistance to imipenem or meropenem and expression levels of efflux pumps of P.aeruginosa.Methods MICs of imipenem or meropenem combined with efflux pump inhibitors including carbonyl cyanide m-chlorophenylhydrazone(CCCP,107 strains)and Phe-Arg-β-naphthylamide(PAβN,71 strains)against imipenem-resistant strains were determined by agar dilution method,and changes of MICs were observed.For 32 strains with different resistant phenotypes to imipenem and meropenem,the mRNA expression levels of three efflux pump genes(mexA,mexD and mexF)were quantified by real time fluorescent quantitative PCR.Results The resistance rate of imipenem and meropenem didn't prove any significant difference in the presence of efflux pump inhibitors.The X2 value of imipenem combined with CCCP and PAβN were 0.338 and 0.086,respectively(P＞0.05),while that of meropenem combined with CCCP and PAβN were 1.065 and 1.458(P＞0.05).No significant in MICs of carbapenems were seen in over half of P. aeruginesa isolates. MICs of carbapenems was significantly downregulated for 4-fold or above in eight isolates. Overexpression of efflux pumps genes were present in 24 of 27 carbapenem-resistant isolates(88. 9% ). Efflux pumps genes including MexAB-OprM, MexCD-OprJ and MexEF-OprN were all overexpressed in 13 isolates,constituting 54. 2% of all carbapenem-resistant isolates. There were 3 isolates in which beth MexAB-OprM and MexCD-OprJ showed overexpression,constituting 12. 5%. Also,MexAB-OprM and MexEF-OprN overexpressed in 3 isolates. There were 2 isolates (8.3%) showing MexEF-OprN overexpression and MexAB-OprM alone. MexCD-OprJ didn't showed overexpression alone. Furthermore,the expression levels of efflux pumps genes mexA,mexD and mexF in isolates susceptible to both in imipenem and meropenem were 0. 48±0. 48,0. 48±0. 53 and 0. 30±0. 41,respectively,which were much lower than that in carbapenem-resistant ones (P＜0. 05 ). MexA gene was expressed at a higher level in meropenemresistant isolates than meropenem-susceptible ones (P＜0. 05 ). Conclusions When the concentration of CCCP and PAβN were 5 μg/ml and 20 μg/ml respectively,the efforts on the carhapenems resistance of P.aeruginosa were small Overexpression of MexAB-OprM might play an important role in meropenemresistance in P. aerugines. Overexpression of MexCD-OprJ and MexEF-OprN was associated with imipenemresistance. However,the relationship between them and meropenem-resistance need to be explored in the future.

EFFECTS OF PODOPHYLLIC ACID PIPERIDYL HYDR-AZONE NITROXIDE RADICAL ON THE PROLIFERATION, MITOTIC INDEX AND DNA SYNTHESIS OF SOME CANCER CELLS IN VITRO / 中国药理学通报

It was found that the proliferation of mice L7712 leukemic cells in vitro was markedly inhibited by 5 mg/L GP1 and VP10 The inhib itory rate increased with the incubation time. At a concentration of 0 .04-20mg/L, the mitotic index ( MI ) of GP1 group increased, but the MI of VP16 group decreased. After L7712 cells were treated with GP, 5 mg/L for 12 h the MI reached the highest point which was 8 times as high as that of the control, at the same time, the MI of VP16 ( 5 mg/L) group was about one-third of that of the control. The result of the combination of GP1 with VP16 showed that VP16 could antagonize the effect of GP on MI of L7712 cells. After being treated by GP1 and VP18 for 24 h, serious damage of L7712 cells could be observed. Both drugs inhibited the incorporation of (3H) TdR into DNA of S180, ascitic hepatoma (AH), L1210 and L7712 cells incubated for 24 h. It was further observed that S180 and L7712 cells were more sensitive than other cells to both drugs.