Synthesis of new N-(3-oxo-1-thia-4-azaspiro[4.5]decan-4-yl)pyridine-3-carboxamide derivatives and evaluation of their anti-influenza virus and antitubercular activities.

We here report the synthesis, structural characterization, and evaluation of the antiviral and antitubercular activities of a novel series of hybrid spirothiazolidinone derivatives (2a-f and 3a-f) containing the nicotinohydrazide moiety, which is an isomer form of the approved antitubercular drug isoniazid. When evaluated for activity against influenza A/H1N1, A/H3N2, and B viruses, three of the new compounds proved to possess specific antiviral activity against the influenza A/H3N2 virus. The most active analog 3a, bearing a 2,8-dimethyl group at the spiro ring, displayed an antiviral EC50 value of 5.2 µM. Compound 3a produced no cytotoxicity at 100 µM, the highest concentration tested, giving a selectivity index of at least 19. Structure-activity relationship analysis indicated that the absence of the methyl substituent at the 2-position and the presence of a bulky substituent at the 8-position of the spirothiazolidinone system caused a significant decrease in antiviral activity. The in vitro antitubercular activity of compounds 2a-f and 3a-f was determined for six different drug-sensitive/drug-resistant laboratory strains and clinical isolates of Mycobacterium tuberculosis. Compounds 2c, 2d, 3b, 3c, and 3d showed weak antitubercular activity against different strains, with MIC values of 125-250 µM.

Evaluation of MALDI-TOF MS for identification of nontuberculous mycobacteria isolated from clinical specimens in mycobacteria growth indicator tube medium.

Nowadays, there is a rising worldwide incidence of diseases caused by nontuberculous mycobacteria (NTM) species, especially in immunocompromised patients and those with underlying chronic pulmonary diseases. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) became a method of choice for the identification of NTM species. The aim of this study was to evaluate MALDI-TOF MS for the identification of NTM isolates compared to the PCR-restriction enzyme analysis (PRA)-hsp65 method. In this study, a total of 152 NTM strains isolated from various clinical specimens were retrospectively analysed. MALDI-TOF MS successfully identified 148 (97.4%) of the 152 NTM isolates but failed to identify four (2.6%) of them. Bruker mycobacteria library gave spectral scores higher than 2.0 for 45 (29.6%) of NTM isolates, between 1.6 and 2.0 for 98 (64.5%) of NTM isolates, and lower than 1.6 for nine (5.9%) NTM isolates. The discordant results between MALDI-TOF MS and PRA-hsp65 analysis were confirmed by sequence analysis. In conclusion, MALDI-TOF MS is a technique capable of performing accurate, rapid, cost-effective, and easy identification of NTM isolates.

[Management and treatment difficulties of multi-drug resistant pulmonary tuberculosis in a pediatric case]. / Çok ilaca direçli akciger tüberkülozu olan çocuk olguda tani ve tedavi yaklasiminda yasanan güçlükler.

Tuberculosis continues to be a major health problem worldwide. Multidrug resistant tuberculosis (MDR-TB) infection that occurs in childhood is caused by adult MDR-TB agents which are in circulation and resistant to primary drugs. In this case report a 17-month-old child with MDR-TB who was cured after a 24-month therapy regimen was presented. Physical examination of a 17-month-old girl admitted to the hospital with the cause of recurrent pneumonia revealed a rubbery lymphadenopathy less than 2 cm in the right upper cervical region. Crepitant rales were detected in the right basal on auscultation of the lung. Interferon gamma release assay (IGRA) and tuberculin skin (TST) tests were negative. Computed tomography (CT) scan of the chest showed mediastinal conglomerate pathologic lymphadenopathy and air bronchograms were detected near the lower lobe of the left lung. Treatment of isoniazid, rifampicin, pyrazinamide with the diagnosis of epituberculosis was started by taking a sample of gastric aspirate culture sample. In the sixth month of the treatment patient was admitted to our clinic with enlarged cervical rubbery lymphadenopathy. It was determined that microbiological test of gastric aspirate culture specimen was positive for M.tuberculosis complex resistant to isoniazid, rifampin, ethambutol, streptomycin, ethionamide and rifabutin. Control CT showed residual peribronchial infiltrations and hilar calcific lymph nodes. Hearing test, vision control and, thyroid function tests were performed and treatment of moxifloxacin, amikacin, para-amino salicylic acid, protionamide and pyrazinamide was started based on minor drug susceptibility results of M.tuberculosis isolate which was still growing in gastric aspirate culture. Gastric aspirate culture for M.tuberculosis was still positive after 3 months of treatment and the current treatment was continued. Amikacin was stopped after 6 months. Therapy regimen was stopped after 24-months. Over the course of a follow-up period of more than 3 years, the clinical and radiological resultsof the patient has improved significantly. The clinical presentation of TB in children is often nonspecific and differs from the patterns seen in adults. MDR-TB cases can be seen in this age group since tuberculosis in children is mainly caused by transmission of drug-resistant strains from adults. This situation is particularly problematic due to the long-term treatment and the lack of specific drug formulations for children.

Indole-based hydrazide-hydrazones and 4-thiazolidinones: synthesis and evaluation as antitubercular and anticancer agents.

A new series of indolylhydrazones (6) and indole-based 4-thiazolidinones (7, 8) have been designed, synthesized and screened for in vitro antitubercular activity against Mycobacterium tuberculosis H37Rv. 4-Thiazolidinone derivatives 7g-7j, 8g, 8h and 8j displayed notable antituberculosis (anti-TB) activity showing 99% inhibition at MIC values ranging from 6.25 to 25.0 µg/ml. Compounds 7g, 7h, 7i, 8h and 8j demonstrated anti-TB activity at concentrations 10-fold lower than those cytotoxic for the mammalian cell lines. The indolylhydrazone derivative 6b has also been evaluated for antiproliferative activity against human cancer cell lines at the National Cancer Institute (USA). Compound 6b showed an interesting anticancer profile against different human tumor-derived cell lines at sub-micromolar concentrations with obvious selectivity toward colon cancer cell line COLO 205.

Prevalence and drug resistance of mycobacteria in Turkish cystic fibrosis patients.

BACKGROUND: Isolation of mycobacteria in cystic fibrosis (CF) patients is increasingly being reported. Because of having long term antimicrobial treatment, CF patients are at risk of pulmonary infection with especially resistant nontuberculous mycobacteria (NTM) strains. The aim of the present study is to determine the prevalence of mycobacterium spp. and antimicrobial susceptibility in Turkish CF patients. METHODS: During a 5.5 year study period, 376 sputa from 130 CF patients were analyzed. Antimycobacterial susceptibility testing was performed by the Bactec 460 TB System and the E test method. RESULTS: Totaly 28 (7.44%) Mycobacterium spp. were isolated from eight (6.15%) CF patients. Five isolates (17.9%) were identified as Mycobacterium tuberculosis complex (MTBC), 14 (50%) as Mycobacterium abscessus and nine (32.1%) as Mycobacterium lentiflavum. All MTBC isolates were found to be susceptible to streptomycin, isoniazid, rifampicin, and ethambutol. Resistance to some antibiotics was detected in some NTM strains. These are the first data about the prevalence of mycobacteria in CF patients from Turkey. CONCLUSIONS: In pediatric CF patients, specific mycobacterial analysis of sputum specimens and susceptibility testing should be performed for allowing early detection, identification and the possibility of eradication of these bacteria.

Synthesis, characterization, and antimicrobial evaluation of some new hydrazinecarbothioamide, 1,2,4-triazole and 1,3,4-thiadiazole derivatives.

In this work, we reported the synthesis and evaluation of antibacterial and antifungal activities of three new compound series obtained from 6-(phenyl/4-chlorophenyl)imidazo[2,1-b]thiazole-3-acetic acid hydrazide: 2-{[6-(phenyl/4-chlorophenyl)imidazo[2,1-b]thiazol-3-yl]acetyl}-N-alkyl/arylhydrazinecarbothioamides (2a-d), 4-alkyl/aryl-2,4-dihydro-5-{[6-(phenyl/4-chlorophenyl)imidazo[2,1-b]thiazol-3-yl]methyl}-3H-1,2,4-triazole-3-thiones (3a-n), and 2-alkyl/arylamino-5-{[6-(phenyl/4-chlorophenyl)imidazo[2,1-b]thiazol-3-yl]methyl}-1,3,4-thiadiazoles (4a-g). The newly synthesized compounds were characterized by IR, (1)H NMR, (13)C NMR (APT), mass and elemental analysis. Their antibacterial and antifungal activities were evaluated against Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Candida albicans ATCC 10231, C. parapsilosis ATCC 22019, C. krusei ATCC 6258, Trichophyton mentagrophytes var. erinacei NCPF 375, Microsporum gypseum NCPF 580, and T. tonsurans NCPF 245. 3c, 3f, 3m, 3n, and 4e showed the highest antibacterial activity. Particularly 3c, 3f, 3g, 3k, 3n, 4a, 4e, and 4g showed the highest antifungal activity against tested fungi.

[Investigation of extensive drug resistance in multidrug resistance tuberculosis isolates]. / Çok Ilaca Dirençli Tüberküloz Izolatlarinda Yaygin Ilaç Direncinin Arastirilmasi

Increasing number of drug resistant tuberculosis (TB) cases, observed in recent years, is an important public health problem. Extensively drug resistant TB (XDR-TB) is the development of resistance against any fluoroquinolones and at least one of the injectable second line anti-TB drugs in addition to resistance against isoniazide and rifampicin which are the first line anti-TB drugs [definition of multidrug resistant TB (MDR-TB)]. Anti-TB therapy failed with first-line anti-TB drugs due to MDR-TB cases is being planned according to second-line anti-TB drug susceptibility test results if available and if not, standart treatment protocols are used. Although it is recommended that individual anti-TB therapy should be designed according to the isolate's susceptibility test results, standart therapeutic protocols are always needed since second-line anti-TB drug susceptibility testing generally could not be performed in developing countries like Turkey. For this reason, nationwide and regional surveillance studies to determine the resistance patterns are always needed to make decisions about the standard therapy algorithms. In this study, it was aimed to investigate the presence of extensive drug resistance among 81 MDR-TB isolates obtained from various health care facilities from Istanbul, Izmir and Manisa and to determine the XDR-TB incidence in Marmara and Aegean regions. Furthermore, we aimed to provide epidemiological data to clinicians to support their choice of second-line anti-TB drugs for MDR-TB infections. Susceptibility testing of isolates for the first and the second-line anti-TB drugs were performed by using modified Middlebrook 7H9 broth in fluorometric BACTEC MGIT 960 system (Becton Dickinson, USA). Eighty-one MDR-TB isolates included in this study were isolated from 43 (53.1%) patients residing in Istanbul, 26 (32.1%) in Izmir and 12 (14.8%) in Manisa provinces. We could not find any isolate consistent with XDR-TB definition in this study. Second-line drug resistance rates of MDR-TB isolates to amikacin and kanamycin were 1.2%, ofloxacin and levofloxacin were 2.5%, capreomycin was 14.8%, ethionamide was 37% whereas linezolid resistance was not detected. Statistically significant correlation was detected between resistance rates of these antibiotic pairs; levofloxacin-ofloxacin (p< 0.01), amikacin-kanamycin (p= 0.01) and streptomycin-ethionamide (p= 0.04). In our study, extensive drug resistance was not encountered in any MDR-TB isolates while high resistance rates was observed against ethionamide and capreomycin. It can be concluded that parenteral aminoglycosides amikasin and kanamycin, fluoroquinolones and linezolid seemed to be reliable anti-TB agents in MDR-TB treatment, however, further larger scale studies are needed.

[Antibiotic susceptibility rates and beta-lactam resistance mechanisms of Pseudomonas aeruginosa strains]. / Pseudomonas aeruginosa Suslarinda Antibiyotik Duyarlilik Oranlari ve Beta-Laktam Direnç Mekanizmalarinin Tiplendirilmesi.

Pseudomonas aeruginosa is a well-known cause of severe and potentially life-threatening infections including bacteremia, skin and wound infections, pulmonary disease, especially among individuals with cycstic fibrosis, nosocomial urinary tract infections, endocarditis and meningitis. The mechanism of resistance to broad-spectrum beta-lactams in P.aeruginosa are overexpression of cephalosporinases and/or class A, B and D beta-lactamases. Recently PER-1 type beta-lactamase has been reported from Turkey, France, Italy, Romania, Hungary, Belgium, Russia, South Korea and India. OXA beta-lactamases have increasingly been reported in clinical strains of P.aeruginosa from various geographical origins. This study was aimed to investigate the antibiotic susceptibility of various P.aeruginosa clinical strains and to define the beta-lactamase enzymes leading to resistance. In this study, a total of 100 P.aeruginosa strains isolated from various clinical specimens (37 urine, 21 blood, 10 sputum, 5 bronchoalveolar lavage, 5 abscess, 5 wound swabs, 4 endotracheal aspirate, 3 throat swabs, 2 catheter tips, one of each pleural and peritoneal fluid) were included. According to Clinical and Laboratory Standards Institute (CLSI) recommendations, susceptibilities of isolates to various antibiotics were investigated by disk diffusion and agar dilution method, and beta-lactamase enzymes were detected by isoelectric focusing (IEF) method. PSE, PER-1, OXA-10-like beta-lactamase genes and MEX-R genes of isolates were investigated by polymerase chain reaction (PCR). According to MIC90 values, the most effective antibiotics were found to be imipenem (8 µg/ml). The MIC90 values of amikacin, ciprofloxacin, cefepime, cefpirome, piperacillin + tazobactam, piperacillin, ceftazidime, ticarcilin, aztreonam and ticarcilin + clavulanic acid were 32, 64, 64, 64, 128/4, 512, 512, 512, 512 and 512/2 µg/ml, respectively. Seven of the isolates were found to be ESBL positive by double-disk synergy method. It was detected that 10% of the isolates were imipenem-susceptible and 9% were intermediate susceptible. Phenotypical investigation of metallo-beta-lactamase enzyme in these strains by MBL E-test method did not reveal a positive result. PER-1 and OXA-10 like beta-lactamases were detected each in 11% of the isolates, and co-presence of PER-like and OXA-10 like enzymes were shown in 4% of the isolates. PSE gene was not found in any of the strains. The MEXR gene was identified in 52% of the isolates. Antibiotic resistance mechanisms in P.aeruginosa strains seems to be complex. Determination of the resistance mechanisms and antibiotic susceptibility rates in P.aeruginosa will guide the proper antimicrobial therapy, reducing the emergence of resistant strains.

Testing susceptibility of multidrug-resistant Mycobacterium tuberculosis to second-line drugs by use of blood agar.

In this study, the susceptibilities of 35 multidrug-resistant (MDR) Mycobacterium tuberculosis clinical isolates to second-line drugs, including kanamycin (KM), rifabutin (RBU), ofloxacin (OFX), p-aminosalicylic acid (PAS), capreomycin (CAP), clofazimine (CFM), and ethionamide (ETH), were investigated on blood agar according to CLSI recommendations. Compared with the results of the Bactec 460 TB system, agreement was 100, 100, 97, 100, 100, 100, and 86% for KM, RBU, OFX, PAS, CAP, CFM, and ETH, respectively. Compared with the results of the proportion method, agreement was 100, 100, 97, 100, 97, 100, and 77% for KM, RBU, OFX, PAS, CAP, CFM, and ETH, respectively.

Synthesis, structure, and antifungal evaluation of some novel 1,2,4-triazolylmercaptoacetylthiosemicarbazide and 1,2,4-triazolylmercaptomethyl-1,3,4-thiadiazole analogs.

Novel 1-[[4-(4-bromophenyl)-5-(2-furyl)-4H-1,2,4-triazole-3-yl]mercaptoacetyl]-4-alkyl/aryl-3-thiosemicarbazides (5-12) were synthesized by the reaction of 4-(4-bromophenyl)-5-(2-furyl)-4H-1,2,4-triazole-3-ylmercaptoacetylhydrazide (4) with substituted isothiocyanates. Cyclodehydration of thiosemicarbazides with concentrated sulfuric acid yielded 2-[4-(4-bromophenyl)-5-(2-furyl)-4H-1,2,4-triazole-3-yl]mercaptomethyl-5-alkyl/arylamino-1,3, 4-thiadiazoles (13-17). The new compounds were evaluated for in vitro antifungal activity using the microdilution method. The tested compounds showed varying degrees of activity against Microsporum gypseum NCPF-580, Microsporum canis, Trichophyton mentagrophytes, Trichophyton rubrum, and Candida albicans ATCC 10231 (MIC 8-4 microg/mL).

Evaluating the effectiveness of anti-tuberculosis treatment by detecting Mycobacterium tuberculosis 85B messenger RNA expression in sputum.

BACKGROUND: The antigen 85 complex (85B) is secreted in large quantities from growing mycobacteria and the presence of bacterial mRNA is an indicator of cell viability. The quantitative detection of 85B mRNA expression levels can be used to assess the success of anti-tuberculosis treatment outcomes to detect viable mycobacteria cells. Therefore, we evaluated the levels of 85B mRNA of Mycobacterium tuberculosis strains in patients with pulmonary tuberculosis. METHODS: Thirty patients with primary tuberculosis were included in this study. The sputum specimens of patients were collected on days 0, 15, and 30 days and were cultured and evaluated by 85B mRNA-based RT-qPCR. RESULTS: Overall, 23 of the studied tuberculosis strains were susceptible to the primary anti-tuberculosis antibiotics used in this study, 7 were resistant. By the 30th day of treatment, 85B mRNA was detected in only one of the susceptible strains, but in all 7 of the resistant strains, though the relative gene expression varied between the strains. This difference between the susceptible and resistant strains at day 30 was statistically significant (p < 0.05). CONCLUSION: 85B mRNA expression levels could be used to follow up on primary tuberculosis cases. 85B mRNA seems to be a good diagnostic marker for monitoring anti-tuberculosis treatment outcomes.

Design, synthesis, antitubercular and antiviral properties of new spirocyclic indole derivatives.

ABSTRACT: A series of indole-based spirothiazolidinones have been designed, synthesized and evaluated, in vitro, for their antitubercular, antiviral, antibacterial, and antifungal activities. The structures of the new compounds were established by IR, 1H NMR, 13C NMR (proton decoupled, APT, and DEPT), electrospray ionization mass spectrometry, and microanalysis. Compounds bearing a phenyl substituent at position 8 of the spiro ring, exhibited significant antitubercular activity against Mycobacterium tuberculosis H37Rv ATCC 27294 at concentrations of 3.9 and 7.8 µM. Still, some of the tested compounds displayed activity on mycobacteria with MIC values of 16 and 31 µM. Four of the indole-spirothiazolidinone derivatives were found to be moderately active against Punta Toro virus, yellow fever virus or Sindbis virus in Vero cells. The antiviral EC50 values were in the range of 1.9-12 µM and the selectivity index (ratio of cytotoxic to antivirally effective concentration) was above 10 in some cases. The most potent effect was seen with the compound that is methylated at positions 2 and 8 of the spirothiazolidinone system.

Synthesis and antimicrobial evaluation of new 3-alkyl/aryl-2-[((alpha,alpha-diphenyl-alpha-hydroxy)acetyl)hydrazono]-5-methyl-4-thiazolidinones.

New 4-thiazolidinone derivatives of benzilic acid (alpha,alpha-diphenyl-alpha-hydroxyacetic acid) have been synthesized and evaluated for antibacterial and antifungal activities. The reaction of 1- (alpha,alpha-diphenyl-alpha-hydroxy)acetyl-4-alkyl/arylthiosemicarbazides with ethyl 2-bromopropionate gave 3-alkyl/aryl-2-[((alpha,alpha-diphenyl-alpha-hydroxy)acetyl)hydrazono]-5-methyl-4-thiazolidinone derivatives. Their antibacterial and antifungal activities were evaluated against S. aureus ATCC 29213, P. aeruginosa ATCC 27853, E. coli ATCC 25922, C. albicans ATCC 10231, C. parapsilosis ATCC 22019, C. krusei ATCC 6258, T. mentagrophytes var. erinacei NCPF 375, M. gypseum NCPF 580 and T. tonsurans NCPF 245. 3e, 3f, 3g and 3h showed the highest antibacterial activity. Particularly 3a and 3e showed the highest antifungal activities against C. parapsilosis ATCC 22019, T. tonsurans NCPF 245 and M. gypseum NCPF 580.