Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: A Faropenem, Linezolid, and Moxifloxacin Regimen for Both Drug-Susceptible and Multidrug-Resistant Tuberculosis in Children: FLAME Path on the Milky Way

doi: 10.1093/cid/ciw474

Figure Lengend Snippet: Exposure-effect relationship of faropenem and Mycobacterium tuberculosis ( Mtb ) on day 7 using inhibitory sigmoid maximal kill (E max ) model. Treatment with static concentrations of faropenem for 7 days resulted in a E max of 2.71 log 10 colony-forming units (CFU)/mL with an exposure mediating 50% of E max of 11.09 mg/L ( r 2 = 0.91), as shown in the equation, which is a kill rate similar to, and better than, most currently used antituberculosis agents.

Article Snippet: Faropenem sodium hydrate powder was purchased from BOC Sciences (Shirley, New York).

Techniques:

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: A Faropenem, Linezolid, and Moxifloxacin Regimen for Both Drug-Susceptible and Multidrug-Resistant Tuberculosis in Children: FLAME Path on the Milky Way

doi: 10.1093/cid/ciw474

Figure Lengend Snippet: Faropenem exposure-response in combination regimen. A , Effect of different faropenem % of time concentration persists above minimum inhibitory concentration (%T MIC ) and linezolid-moxifloxacin regimen on THP-1 cells. There was no effect of faropenem exposures on THP-1 cells. B , Effect of different faropenem exposures in combination with the linezolid-moxifloxacin regimen based on an exponential decline model for Mycobacterium tuberculosis ( Mtb ) log 10 colony-forming units (CFU)/mL. The faropenem T MIC of 62.5% had a steeper slope than 12.5% and 25%. There was no convergence for the exponential decline model for nontreated controls, as expected, given they did not decline.

Article Snippet: Faropenem sodium hydrate powder was purchased from BOC Sciences (Shirley, New York).

Techniques: Concentration Assay

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: A Faropenem, Linezolid, and Moxifloxacin Regimen for Both Drug-Susceptible and Multidrug-Resistant Tuberculosis in Children: FLAME Path on the Milky Way

doi: 10.1093/cid/ciw474

Figure Lengend Snippet: Microbial responses to optimized faropenem-based regimens in a paucibacillary model. A , The experimental faropenem, linezolid, and moxifloxacin (FLM) regimen and high-dose moxifloxacin (FLM Hi ) plus ethambutol (EMB), as well as the standard therapy regimen, had killed Mycobacterium tuberculosis ( Mtb ) in the pediatric hollow fiber system model for intracellular tuberculosis systems below colony-forming unit (CFU)/mL assay limits, suggesting complete sterilization. B , The time to positivity (TTP) assay revealed that there was still bacterial growth with the FLM regimen and standard regimen; the former has statistically higher TTP compared with standard regimen at the end of experiment, suggesting better kill rates.

Article Snippet: Faropenem sodium hydrate powder was purchased from BOC Sciences (Shirley, New York).

Techniques:

Journal: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America

Article Title: A Faropenem, Linezolid, and Moxifloxacin Regimen for Both Drug-Susceptible and Multidrug-Resistant Tuberculosis in Children: FLAME Path on the Milky Way

doi: 10.1093/cid/ciw474

Figure Lengend Snippet: Effect of a new faropenem, linezolid, and moxifloxacin (FLM) regimen over 1 month. A , Kill slopes of the different regimens based on a linear regression model and 95% confidence bounds. The r 2 for the FLM regimen was 0.92, for high-dose moxifloxacin (FLM Hi ) was 0.84, for FLM Hi with ethambutol (EMB) was 0.84, and for standard therapy was 0.73. B , Exponential decline model. The decline rates of all the regimens were similar to standard therapy. C , Time to positivity (TTP) of contents from each hollow fiber system shows that there is no sterilization up to day 28 with TTP when compared to Mycobacterium tuberculosis ( Mtb ) log colony-forming units (CFU)/mL, which revealed sterilization. In this experiment we started with a higher bacterial burden, an acid test. Results of multiple t test comparison shown as * P value of .045 on day 7 and ** P = .011 for day 21. However, by day 28 the TTPs were similar in all drug treatment regimens.

Article Snippet: Faropenem sodium hydrate powder was purchased from BOC Sciences (Shirley, New York).

Techniques: Comparison

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Biochemical characterization of β-lactam activity. Spectrophotometric determination of kinetic constants for LdtMt1 inactivation by faropenem and meropenem (A to C) and for hydrolysis of these antibiotics by BlaC (D to F). The first-order constants (kobs) were determined for six drug concentrations by measuring the absorbance decrease (30-s time scale), which results from opening of the antibiotic β-lactam ring upon enzyme acylation, and regression analysis with the equation [EI*] = [Etotal](1 − e−kobst), in which [Etotal] is the total enzyme concentration, kobs is a constant, and t is time. The resulting kobs values were plotted as a function of antibiotic concentrations (A), and kinetic constants kinact and Kapp (C) were deduced by regression analysis using the equation kobs = kinact[I]/(Kapp + [I]), where [I] is the antibiotic concentration, kinact is the first-order rate constant for LdtMt1 acylation, and Kapp is a constant. The hydrolysis velocities of faropenem and meropenem (1,000-min time scale) were plotted as a function of the LdtMt1 concentration (B), and enzyme turnover (khydrol) was deduced from the slope (C). This provides an estimate of the rate of hydrolysis of the acyl enzyme since this is the rate-limiting step of the full catalytic cycle. EIox, oxyanion. (D to F) Determination of kinetic constants for hydrolysis of faropenem and meropenem by β-lactamase BlaC. Turnover numbers were determined for various concentrations of meropenem (D) and faropenem (E), and regression analysis was performed using the equation turnover number = kcat[S]/(Km + [S]) (F), where [S] is the antibiotic concentration.

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Activity Assay, Concentration Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Activity of faropenem against extracellular and intracellular M. tuberculosis. (A) Bacteria expressing luciferase were incubated for 7 days in 7H9 medium containing different concentrations of faropenem (FPM) or faropenem plus clavulanate (FPMC). Growth was measured using a luciferase-based assay, and the data were normalized to the values obtained in the absence of antibiotic. Results are representative of those from four experiments. (B) Log-phase cultures were exposed for 6 days to different concentrations of faropenem and plated to measure the number of CFU. Values were normalized to those for the untreated controls. Results are representative of those from two experiments. (C) Log-phase cultures were exposed to antibiotics with (filled symbols) or without (empty symbols) 2.5 μg/ml clavulanate. Circles, meropenem (8 μg/ml; ∼26× MIC with clavulanate, ∼3× MIC without clavulanate); squares, faropenem (8 μg/ml; ∼6× MIC with or without clavulanate); triangles, faropenem (28 μg/ml; ∼21× MIC); crosses, 0.5 μg/ml isoniazid (∼20× MIC). At 0, 1, 2, 4, and 8 days after antibiotic addition, aliquots were washed and plated to measure the number of CFU. Results are means ± SEMs from three experiments. The rebound of the numbers of CFU at 8 days in the culture exposed to isoniazid was due to the outgrowth of drug-resistant variants. (D) The stability of compounds in 7H9 medium was measured by HPLC-UV analysis. Clav, clavulanic acid; MPM, meropenem. (E and F) Bacteria expressing GFP were grown in RAW macrophages and left untreated (closed circles) or treated with 56 μg/ml faropenem (open squares), 50 μg/ml meropenem plus 2.5 μg/ml clavulanate (open circles), or 0.75 μg/ml isoniazid (crosses). (E) Macrophage lysates were plated at 0 h and at 1, 3, and 6 days postinfection to measure the number of CFU. Data are plotted as percent survival normalized to the number of organisms at 0 h. (F) The fraction of macrophages (Mϕ) containing GFP-positive M. tuberculosis was measured by flow cytometry at 1, 3, and 6 days postinfection. INH, isoniazid; FPM, faropenem; MPMC, meropenem plus clavulanate. Results are means ± SEMs from three experiments.

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Activity Assay, Bacteria, Expressing, Luciferase, Incubation, Flow Cytometry

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Real-time single-cell analysis of faropenem-mediated growth inhibition. An M. tuberculosis strain expressing GFP was cultured in a microfluidic device under a constant flow of 7H9 medium and imaged at 1-h intervals. (A and D) Faropenem (28 μg/ml; ∼21× MIC) was added to the flow medium at 96 to 264 h. (B and E) Meropenem plus clavulanate (MPC; 8 μg/ml and 2.5 μg/ml, respectively; ∼26× MIC) was added to the flow medium at 96 to 330 h. (C and F) Isoniazid (0.5 μg/ml; ∼20× MIC) was added to the flow medium at 96 to 340 h. (A to C) Images were recorded on fluorescence (green) and phase channels and merged. Numbers (lower right) indicate the elapsed times (in hours). Labels (upper right) indicate the presence or absence of antibiotic in the flow medium. (D to F) Single-cell growth rates were determined by measuring the projected areas of single cells (n = 50) during the indicated time intervals and fitting exponential curves to the data.

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Single-cell Analysis, Inhibition, Expressing, Cell Culture, Fluorescence

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Real-time single-cell analysis of faropenem-mediated cytolysis of uropathogenic E. coli. Uropathogenic E. coli CFT073 expressing YFP was cultured in a microfluidic device under a constant flow of LB medium and imaged at 2-min intervals. Faropenem (4 μg/ml; ∼20× MIC) was added to the flow medium at 2 to 8 h. (A) Images were recorded on the fluorescence (green) and phase channels and merged. Numbers (lower right) indicate the elapsed times (in hours). Labels (upper right) indicate the flow medium. Scale bars, 5 µm. (B) Single-cell growth rates were determined by measuring the projected areas of single cells (n = 50) during the indicated time intervals and fitting exponential curves to the data. NS, no significant difference.

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Single-cell Analysis, Expressing, Cell Culture, Fluorescence

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Real-time single-cell analysis of faropenem-mediated cytolysis. An M. tuberculosis strain expressing GFP was cultured in a microfluidic device under a constant flow of 7H9 medium and imaged at 1-h intervals. (A and D) Faropenem (28 μg/ml; ∼21× MIC) was added to the flow medium at 96 to 264 h. (B and E) Meropenem plus clavulanate (8 μg/ml and 2.5 μg/ml, respectively; ∼26× MIC) was added to the flow medium at 96 to 330 h. (C and F) Isoniazid (0.5 μg/ml; ∼20× MIC) was added to the flow medium at 96 to 340 h. Cytolysis was scored visually as an abrupt loss of GFP fluorescence and an abrupt decrease in phase intensity. As an endpoint assay, 1.0 μg/ml PI was added to the flow medium for 24 h to stain cells with permeabilized cell envelopes. (A to C) Number of intact cells plotted for individual (left) and ensemble (right) microcolonies exposed to faropenem (n = 1,593 cells in 50 microcolonies) (A), meropenem plus clavulanate (n = 1,541 cells in 65 microcolonies) (B), or isoniazid (n = 1,917 cells in 41 microcolonies) (C). Shading, the interval of antibiotic exposure; red lines, curve fitting of the data using a two-phase decay exponential equation. (D to F) Percentage of single cells scored in three categories: lysed (white), intact but PI positive (red), and intact and PI negative (green).

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Single-cell Analysis, Expressing, Cell Culture, Fluorescence, End Point Assay, Staining

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: MICs against Mycobacterium tuberculosis H37Rv

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Kinetic constants for inactivation of Ldt Mt1 to Ldt Mt5 by β-lactams a

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques:

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Efficacy of faropenem in a mouse model of tuberculosis. C57BL/6J mice infected with M. tuberculosis were treated with 500 mg/kg faropenem medoxomil (F), 25 mg/kg clavulanate plus 200 mg/kg probenecid (C+P), or 500 mg/kg faropenem medoxomil plus 25 mg/kg clavulanate and 200 mg/kg probenecid (F+C+P) on days 1 to 8 postinfection. The bacterial loads in lungs harvested on day 9 were determined, and the fold change in the number of CFU relative to the number of CFU for the untreated controls (UNT) is depicted. Results are means ± SEMs for five mice in each group. *, P < 0.05, Mann-Whitney test.

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques: Infection, MANN-WHITNEY

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

doi: 10.1128/AAC.03461-14

Figure Lengend Snippet: Pharmacokinetic parameters of faropenem in mice a

Article Snippet: The kinetic constants for Ldt Mt1 acylation by faropenem (sodium hemipentahydrate; GlaxoSmithKline) and meropenem (trihydrate; AstraZeneca) were determined in 100 mM sodium phosphate (pH 6.0) by measuring the decrease in the absorbance at 299 nm, which results from opening of the antibiotic β-lactam ring (Δε 299 for faropenem and meropenem = −5,000 and −7,100 M −1 cm −1 , respectively).

Techniques:

Journal:

Article Title: In Vitro and In Vivo Antibacterial Activities of L-084, a Novel Oral Carbapenem, against Causative Organisms of Respiratory Tract Infections

doi: 10.1128/AAC.45.1.203-207.2001

Figure Lengend Snippet: Comparative in vitro activities of L-036 and reference drugs against clinical isolates

Article Snippet: The following antimicrobial agents, used in this study, were obtained from the indicated sources: L-036 and L-084, Wyeth Lederle Japan, Tokyo, Japan; faropenem, Suntory, Tokyo, Japan; imipenem, Banyu Pharmaceutical Co., Tokyo, Japan; levofloxacin, Daiichi Pharmaceutical Co., Tokyo, Japan; cefditoren and cefditoren-pivoxil, Meiji Seika Co., Tokyo, Japan; cefdinir and amoxicillin, Fujisawa Pharmaceutical Co., Osaka, Japan.

Techniques: In Vitro

Journal:

Article Title: In Vitro and In Vivo Antibacterial Activities of L-084, a Novel Oral Carbapenem, against Causative Organisms of Respiratory Tract Infections

doi: 10.1128/AAC.45.1.203-207.2001

Figure Lengend Snippet: Therapeutic efficacy of L-084 and reference drugs in murine respiratory tract infections caused by penicillin-susceptible S. pneumoniae TUH39 a

Article Snippet: The following antimicrobial agents, used in this study, were obtained from the indicated sources: L-036 and L-084, Wyeth Lederle Japan, Tokyo, Japan; faropenem, Suntory, Tokyo, Japan; imipenem, Banyu Pharmaceutical Co., Tokyo, Japan; levofloxacin, Daiichi Pharmaceutical Co., Tokyo, Japan; cefditoren and cefditoren-pivoxil, Meiji Seika Co., Tokyo, Japan; cefdinir and amoxicillin, Fujisawa Pharmaceutical Co., Osaka, Japan.

Techniques:

Journal:

Article Title: In Vitro and In Vivo Antibacterial Activities of L-084, a Novel Oral Carbapenem, against Causative Organisms of Respiratory Tract Infections

doi: 10.1128/AAC.45.1.203-207.2001

Figure Lengend Snippet: Therapeutic efficacy of L-084 and reference drugs against murine respiratory tract infection caused by penicillin-resistant S. pneumoniae TUM741

Article Snippet: The following antimicrobial agents, used in this study, were obtained from the indicated sources: L-036 and L-084, Wyeth Lederle Japan, Tokyo, Japan; faropenem, Suntory, Tokyo, Japan; imipenem, Banyu Pharmaceutical Co., Tokyo, Japan; levofloxacin, Daiichi Pharmaceutical Co., Tokyo, Japan; cefditoren and cefditoren-pivoxil, Meiji Seika Co., Tokyo, Japan; cefdinir and amoxicillin, Fujisawa Pharmaceutical Co., Osaka, Japan.

Techniques: Infection

Journal:

Article Title: In Vitro and In Vivo Antibacterial Activities of L-084, a Novel Oral Carbapenem, against Causative Organisms of Respiratory Tract Infections

doi: 10.1128/AAC.45.1.203-207.2001

Figure Lengend Snippet: Therapeutic efficacy of L-084 and reference drugs against murine respiratory tract infection caused by H. influenzae TUM8

Article Snippet: The following antimicrobial agents, used in this study, were obtained from the indicated sources: L-036 and L-084, Wyeth Lederle Japan, Tokyo, Japan; faropenem, Suntory, Tokyo, Japan; imipenem, Banyu Pharmaceutical Co., Tokyo, Japan; levofloxacin, Daiichi Pharmaceutical Co., Tokyo, Japan; cefditoren and cefditoren-pivoxil, Meiji Seika Co., Tokyo, Japan; cefdinir and amoxicillin, Fujisawa Pharmaceutical Co., Osaka, Japan.

Techniques: Infection