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Biapenem (biapenem) is a new carbapenem antibiotic, antibacterial mechanism play an antibacterial effect by inhibiting bacterial cell wall synthesis, broad spectrum antimicrobial, antibacterial activity against gram-negative and gram-positive need oxygen and anaerobic bacteria and so has an antibacterial effect and fewer other β-lactam antibiotics common bacterial resistance. Injection in clinical Biapenem applicable to the treatment of sepsis caused by susceptible bacteria, pneumonia, lung abscess, chronic respiratory disease caused by secondary infections refractory cystitis, pyelonephritis, peritonitis, gynecological accessories go far . Injection Biapenem of human renal dehydropeptidase-I (DHP-I) stable, can be administered alone, without the need to be combined with the DHP-I inhibitor; drugs can be effectively through the blood-brain barrier can be in clinical used in the treatment of patients with meningitis; stimulate the central nervous system induced epilepsy is very unlikely. Objective To optimize human plasma HPLC method Biapenem in, to establish a highly efficient, sensitive, loyal, the convenient human plasma drug concentration detection methods in Biapenem, and a factory in some sort of preparation process conditions evaluate the quality of the production of injection Biapenem to the different formulations environment proved different manufacturers, different preparation process of the case, the regularity and characteristics of the disposal of the drug in the body produced by Phase I clinical data for the completion of the drug's clinical reporting , to provide a basis for a Phase II clinical trial, provide the basis for clinical application. This study is divided into the following two parts the first part of the optimization of sample processing and detection methods to inspect selected specificity, high sensitivity, good accuracy and high performance liquid chromatography. Detection step: precision draw 200μL plasma, adding internal standard solution 20μL0.1 mg / mL, vortex 10 s, to add 100μL 7% aqueous solution of zinc sulfate, vortex 3 min, mixing, in of 16000r · min - 1 centrifuge 5 min, the supernatant injection analysis, the injection volume of 20 μL. The second part of the study of single and multiple intravenous infusion injection the pharmacokinetics after Biapenem dynamic characteristics. 30 healthy subjects randomized parallel divided into three groups, each group of 10 healthy subjects (male and female and female), low-dose group single intravenous infusion of 150mg; middle dose group single intravenous infusion of 300 mg; a single intravenous infusion of the high dose group of 600 mg; diluted with 0.9% sodium chloride injection, the total volume of the infusion of the subjects are 100mL, intravenous infusion time of 1 h, 0 h before the administration began administered 15 min, 30 min, 45 min, 1.0 h after the administration, 30 min, 1.0 h, 1.5 h, 2.0 h and 2.5 h, 4.0 h, 6.0 h taken by the other side of the forearm vein blood 4 ml / person / times, set sterile heparin tubes, centrifugation plasma set sterile tubes, and stored at -20 ℃. Multiple dosing middle dose group of subjects after the single dose test, at a dose of 300 mg / (liquid dilution method with single dose), one day, administered for 5 consecutive days each infusion time are 1.0 h (single dose count one day, five days administered once only). Morning pre-dose at day 2, 3, 4, 5, and 5 morning administration after administration for 30 min, 1.0 h after the administration, 15 min, 30 min, 1.0 h, 1.5 h, 2.0 h, 2.5 H, 3.0 h, 4.0 h, 6.0 h, 8.0 h forearm vein blood 4 ml / heparin tubes, separating the plasma, -20 ℃ stored analyte. Concentration Biapenem plasma were determined by HPLC, and pharmacokinetic parameters were calculated with DAS2.0. Results of this test to establish the plasma injection with Biapenem HPLC method, the impurities in the plasma does not interfere with the determination of the sample; standard curve was linear over the range of 0.2μg · mL -1 -50μg · mL -1 , a good linear relationship; minimum detection limit was 0.2μg · mL -1 ; high, medium, and low concentration intra and inter-assay precision degrees RSD were less than 15.0%; extraction recoveries of 73% -75%; determination requirements of biological samples. 2.30 healthy subjects, a single intravenous infusion of different doses of injection with Biapenem (150 mg, 300 mg, 600 mg) in vivo pharmacokinetic parameters were: Tmax: 0.93 ± 0.12 h, 1.01 ± 0.16h, 1.00 ± 0.00h, Cmax: 8.92 ± 4.13mg · L -1 , 19.07 ± 3.08mg · L -1 , 28.01 ± 6.95mg · L -1 ; t1 / 2:1.47 ± 0.23h, 0.98 ± 0.26h, 1.90 ± 0.53h; AUC0-t: 11.24 ± 3.09mg · h · L -1 , 22.15 ± 3.96mg · h · L -1 , 35.58 ± 4.65mg · h · L -1 ; AUC0-∞: 12.01 ± 3.12mg · h · L -1 , 22.51 ± 3.99mg · h · L -1 , 37.27 ± 5.01mg · h · L -1 ; CL / F: 0.013 ± 0 -003L · h -1 , 0.007 ± 0. 001L · h -1 , 0.016 ± 0.002L · h -1 . 10 subjects were repeatedly intravenously injected with Biapenem, 300 mg / day, administered for 5 consecutive days twice in the administration of the second day the injection Biapenem reached steady-state plasma concentration, the steady-state average plasma concentration of 0.68 ± 0.11 mg · L -1 , steady-state plasma concentration fluctuation was 5.81 ± 1.65, accumulation constant of 0.96 ± 0.02. After repeated administration to the data to estimate the continuous administration of the blood concentration of the first 5 days of the injection with biapenem elimination half-life of 1.68 ± 0.37 h, AUC0-t 24.69 ± 2.81 mg · h · L - 1 , the time to peak and peak concentrations were 1.00 ± 0.00 h and 18.97 ± 6.04 mg · L -1 . The conclusions of the experiment to establish human plasma Biapenem HPLC method precision, accuracy, selectivity, good reproducibility. The plasma Biapenem concentration 0.2-50 μg · mL -1 range, a good linear relationship, r 0.999. Detection wavelength of 300 nm, the average retention time of the sample peak is about 3.95min minimum detection limit was 0.2μg · mL -1 , high, low concentrations of the intraday precision coefficient of variation were less than 5%, and extraction recoveries were more than 75%. All performance indicators are in line with the test requirements for the pharmacokinetic study. The plasma concentration of subjects by recorded in the DAS2.0 program, choose the best weight compartment model subjects batch data analysis by dose group, 30 healthy volunteers dose intravenous infusion Biapenem, low, three high dose group elimination half-life t1 / 2 were (1.15 ± 0.17), (1.06 ± 0.28), (1.05 ± 0.06) h, three dose groups t1 / 2 are basically the same show that was an elimination kinetics than A Peinan 150-600 mg; low dose group, three high Cmax were (8.92 ± 0.41), (19.07 ± 3.08) and ( 35.60 ± 6.95) mg · L -1 ; AUC0-t were (11.17 ± 3.01), (22.14 ± 3.97), (45.26 ± 3.85) mg · L -1 · h, Cmax and AUC0-t and the dose was linearly correlated (P lt; 0.001) that was linear pharmacokinetic characteristics than A Peinan 150-600 mg, regression equations were Cmax = 0.0587X-0.655 (R2 = 0.9972) and AUC0-th = 0.0759X-0.39 (R2 = 0.9998). Multiple dosing t1 / 2, Cmax, AUC0-t were (1.21 ± 0.13) h, (22.14 ± 3.97) mg · L -1 · h, (18.96 ± 6.03) mg · L -1 · h multiple dosing compared with single-dose pharmacokinetic data no significant differences, suggesting that in vivo than A Peinan no accumulation. 3. Consistent pharmacokinetic parameters of this study the pharmacokinetic parameters of foreign literature study, no adverse reactions showed that injection Biapenem good security at this concentration.
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