X-ray crystallographic analysis of IMP-1 metallo-ß-lactamase complexed with a 3-aminophthalic acid derivative, structure-based drug design, and synthesis of 3,6-disubstituted phthalic acid derivative inhibitors.

3-(4-Hydroxypiperidine-1-yl) phthalic acid 1 shows potent inhibitory activity against metallo-ß-lactamase, which is known to inactivate ß-lactam antibiotics such as carbapenems. Here, the structure of co-crystals of the metallo-ß-lactamase IMP-1 and 1 was first analyzed by X-ray crystallography, and then used for structure-based drug design. Four novel compounds bearing substituents at the 6-position were synthesized to produce 3,6-disubstituted phthalic acid derivatives, and their IMP-1 inhibitory activity and synergistic effect with the carbapenem biapenem (BIPM) were evaluated. 3,6-Disubstituted phthalic acid derivatives showed potent IMP-1 inhibitory activity. In particular, compound 13 showed 10-fold higher IMP-1 inhibitory activity as compared with the parent derivative 1.

Metallo-ß-lactamase inhibitory activity of 3-alkyloxy and 3-amino phthalic acid derivatives and their combination effect with carbapenem.

3-Alkyloxy and 3-amino phthalic acid derivatives were found to have metallo-ß-lactamase inhibitory activity. Among them, 3-amino phthalic acid derivatives showed both potent activity against metallo-ß-lactamase, IMP-1 inhibitory activity and a strong combination effect with biapenem (BIPM), carbapenem antibiotic. In particular, the 4'-hydroxy-piperidine derivative showed strong IMP-1 inhibitory activity and a combination effect with various antibiotics.

Development of reference equations for spirometry in Japanese children aged 6-18 years.

BACKGROUND: Spirometry is the most widely used pulmonary function test and the measured values of spirometric parameters need to be evaluated using reference values predicted for the corresponding race, sex, age, and height. However, none of the existing reference equations for Japanese children covers the entire age range of 6-18 years. The Japanese Society of Pediatric Pulmonology had organized a working group in 2006, in order to develop a new set of national standard reference equations for commonly used spirometric parameters that are applicable through the age range of 6-18 years. METHODS: Quality assured spirometric data were collected through 2006-2008, from 14 institutions in Japan. We applied multiple regression analysis, using age in years (A), square of age (A(2)), height in meters (H), square of height (H(2)), and the product of age and height (AH) as explanatory variables to predict forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV(1)), peak expiratory flow (PEF), forced expiratory flow between 25% and 75% of the FVC (FEF(25-75%)), instantaneous forced expiratory flow when 50% (FEF(50%)) or 75% (FEF(75%)) of the FVC have been expired. RESULTS: Finally, 1,296 tests (674 boys, 622 girls) formed the reference data set. Distributions of the percent predicted values did not differ by ages, confirming excellent fit of the prediction equations throughout the entire age range from 6 to 18 years. Cut-off values (around 5 percentile points) for the parameters were also determined. CONCLUSIONS: We recommend the use of this new set of prediction equations together with suggested cut-off values, for assessment of spirometry in Japanese children and adolescents.

Metallo-beta-lactamase inhibitory activity of phthalic acid derivatives.

4-Butyl-3-methylphthalic acid was recognized as a metallo-beta-lactamase inhibitor. The structure-activity relationship study of substituted phthalic acids afforded 3-phenylphthalic acid derivatives as potent IMP-1 inhibitors. On the other hand, 3-substituted with 4-hydroxyphenyl phthalic acid derivative displayed a potent combination effect with biapenem (BIPM) against Pseudomonas aeruginosa that produce IMP-1.

Application of a shortened inhaled adenosine-5'-monophosphate challenge in young children using the forced oscillation technique.

BACKGROUND: Adenosine-5'-monophosphate (AMP) is an indirect challenge agent thought to reflect allergic airway inflammation. The forced oscillation technique (FOT) is ideal for use in young children and is suitable for inhaled challenge studies in patients who are in this age group. We assessed the agreement between a shortened and a standard AMP challenge and the repeatability of the shortened AMP challenge using FOT as a primary outcome variable. METHODS: Eighteen children completed a shortened and a standard AMP challenge, and 20 children completed repeated shortened AMP challenges. The children inhaled nebulized AMP tidally for 2 min, following which the presence of wheeze and pulse oximetric saturation (Spo(2)) was recorded prior to FOT measurement. Testing continued until the maximum dose was reached or until wheeze, a decrease in Spo(2) to < 90%, or an increase in respiratory resistance at 8 Hz of 2.0 hPa/s/L or 30% was noted. Concordance was assessed as a binary response, and agreement in provocation concentrations (PCs) causing a response was assessed with intraclass correlations. RESULTS: There was a high degree of concordance between the shortened and standard AMP protocols (94%) and repeated shortened AMP protocols (100%). The mean log(10) PCs displayed a high degree of agreement for both AMP protocols, with intraclass correlation coefficients of 0.94 (95% confidence interval, 0.85 to 0.98) and 0.94 (95% confidence interval, 0.82 to 0.98), respectively. CONCLUSIONS: We demonstrated that a shortened AMP challenge that can be applied to young children is comparable to the standard AMP challenge and is highly repeatable. Further studies in young children to assess the clinical role of a shortened AMP challenge using FOT are required.

Assessment of bronchodilator responsiveness in preschool children using forced oscillations.

BACKGROUND: The forced oscillation technique (FOT) requires minimal patient cooperation and is feasible in preschool children. Few data exist on respiratory function changes measured using FOT following inhaled bronchodilators (BD) in healthy young children, limiting the clinical applications of BD testing in this age group. A study was undertaken to determine the most appropriate method of quantifying BD responses using FOT in healthy young children and those with common respiratory conditions including cystic fibrosis, neonatal chronic lung disease and asthma and/or current wheeze. METHODS: A pseudorandom FOT signal (4-48 Hz) was used to examine respiratory resistance and reactance at 6, 8 and 10 Hz; 3-5 acceptable measurements were made before and 15 min after the administration of salbutamol. The post-BD response was expressed in absolute and relative (percentage of baseline) terms. RESULTS: Significant BD responses were seen in all groups. Absolute changes in BD responses were related to baseline lung function within each group. Relative changes in BD responses were less dependent on baseline lung function and were independent of height in healthy children. Those with neonatal chronic lung disease showed a strong baseline dependence in their responses. The BD response in children with cystic fibrosis, asthma or wheeze (based on both group mean data and number of responders) was not greater than in healthy children. CONCLUSIONS: The BD response assessed by the FOT in preschool children should be expressed as a relative change to account for the effect of baseline lung function. The limits for a positive BD response of -40% and 65% for respiratory resistance and reactance, respectively, are recommended.

Respiratory function in healthy young children using forced oscillations.

BACKGROUND: Monitoring of respiratory function is important in the diagnosis and management of respiratory disease. The forced oscillation technique requires minimal patient cooperation and is ideal for the determination of respiratory function in young children. This study aimed to develop reference ranges and to document the repeatability in healthy young children using commercially available forced oscillation equipment. METHODS: The forced oscillation technique, which uses a pseudo-random noise forcing signal between 4 and 48 Hz, was used to measure respiratory function in healthy young children. Repeatability over a 15 min period was also assessed. Regression equations and standardised Z scores were determined for respiratory resistance (Rrs) and reactance (Xrs) at 6, 8 and 10 Hz. RESULTS: Respiratory function was obtained in 158 healthy children aged two to seven years and between 92 and 127 cm in height. Oscillatory respiratory mechanics exhibited linear relationships with height. Within-test variability for resistance ranged between 6% and 9% and between 17% and 20% for reactance. Resistance and reactance did not change significantly over a 15 min period. CONCLUSIONS: Reference ranges for respiratory impedance variables in healthy children aged two to seven years are presented. The short-term repeatability of forced oscillatory variables in this age group is reported, allowing appropriate cut-off values for therapeutic interventions to be defined.

Efficacy of a novel oral carbapenem, tebipenem pivoxil (TBM-PI), against experimental otitis media caused by penicillin resistant Streptococcus pneumoniae in chinchilla.

An animal model of otitis media using chinchillas was developed to evaluate the efficacy of tebipenem pivoxil (TBM-PI) against experimental otitis media. Chinchillas inoculated via the transbullar approach with Streptococcus pneumoniae serogroup 6 were included in the efficacy study with TBM-PI, amoxicillin (AMX) or untreated as controls. TBM-PI resulted in survival rate of 83%, compared with 25% survival for AMX and 0% survival for controls (p<0.01). Quantitative cultures in the middle ear effusions at day 5 of the TBM-PI group yielded 3.5+/-2.4log(10)CFUs/ml. TBM-PI is a promising antibiotic for the treatment of acute otitis media.

Comparison of activities of beta-lactam antibiotics against Streptococcus pneumoniae with recombinant penicillin-binding protein genes from a penicillin-resistant strain.

We investigated the antibacterial activities of 19 beta-lactams against three recombinant bacterial strains, in which three penicillin-binding protein genes, pbp2x, pbp1a, and pbp2b, from penicillin-resistant Streptococcus pneumoniae (PRSP), were transformed to a penicillin-susceptible strain. By the acquisition of the pbp2x gene from PRSP, the minimum inhibitory concentrations (MICs) of third-generation cephalosporins were increased more than eight fold. When the strain acquired the PRSP pbp1a gene in addition to pbp2x, the MICs of all tested beta-lactams increased 2- to 16-fold. When the strain acquired the PRSP pbp2b gene in addition to pbp2x and pbp1a, the MICs of penicillins and carbapenems increased 4- to 16-fold. However, two novel carbapenems, ME1036 and L-036, showed excellent antibacterial activities against these recombinant strains, as well as against the parent PRSP.