The Pharmacokinetics And Bioequivalence Study Of Zaltoprofen In Human

Zaltoprofen is a potent non-steroidal anti-inflammatory drug and has anti-inflammatory,analgesic, antipyretic efficacy for the treatment of chronic rheumatoid arthritis, osteoarthritis,low back pain, and other pain syndromes after surgery, trauma and tooth extraction.Zaltoprofen has not yet approved for marketing in China, and no imports.The domestic literature has not yet reported the pharmacokinetics of zaltoprofen inhuman. This study is strictly following “Technical guidance for chemical drugs in clinicalpharmacokinetic studies” and “The appendix Ⅱ of Chinese pharmacopoeia”. Primarilythrough exploration and establishment of the LC-MS/MS method to detect zaltoprofen inhuman plasma and urine, application to a single and multiple pharmacokinetic characteristicsin healthy human body. It is the first comprehensive and systematic clarify the laws of safety,absorption, metabolic, distribution, excretion of the new drug in healthy Chinese volunteer,and to provide the evidence of dosage regimen for Phase II clinical trials, and evaluate thesafety of zaltoprofen. This study obtained scientific significance and application values infollowing aspects:1. Establishment of zaltoprofen analysis methodObjective: To establish a rapid, specific and sensitive LC-MS/MS method to detectzaltoprofen in plasma and urine and application to a pharmacokinetics study of zaltoprofen in human.Methods: An Agilent ZORBAX Eclipse Plus-C18column（150mm×4.6mm,5μm） wasused with the mobile phase of methanol and0.1%formic acid（90:10, V:V）. Diazepam wasused as internal standard. Multiple reaction monitoring （MRM） mode was employed, and thetransition of m/z was m/z299.2�'m/z225.2for zaltoprofen, m/z285.1�'m/z193.1fordiazapan. The plasma and urine samples were simply pretreated by protein precipitation.Result: The established LC-MS/MS method in human plasma and urine were directly proteinprecipitation, the sample handling simplified, the sample analysis time is shorter. The linearrange of zaltoprofen in plasma is0.02-20μg·mL^-1, and the limit of quantification is0.02μg·mL^-1. RSD of the inter-day and intra-day precisions of low, medium and highconcentrations were all less than15%. Absolute recovery is greater than85%. The linearrange of zaltoprofen in urine is0.1-50μg·mL^-1, RSD of the inter-day and intra-day precisionsof low, medium and high concentrations were all less than15%. Absolute recovery is greaterthan95%. The results of medium effect and stability in biological samples are in line withthe requirements of determination.Conclusion: The simple, specific and rapid LC-MS/MS method for analysis of zaltoprofenin human plasma and urine has been developed and validated, and suitable of thepharmacokinetics study of zaltoprofen.2. Single-dose pharmacokinetics study of zaltoprofenObjective: To detect zaltoprofen in plasma and urine and evaluate the single doseparmacokinetic characteristics for absorption, distribution and eliminate of zaltoprofen inhuman.Methods:30healthy volunteers are divided into three groups randomly. Every group has10volunteers （Male:5, Female:5）. Single oral dose is80mg,160mg,240mg zaltoprofentablets. Venous blood samples were collected at0（pre-dose） and0.5,0.75,1.0,1.5,2.0,3.0,4.0,5.0,6.0,8.0,10.0,12.0and24.0h post-dose. Venous blood4ml were collected, placed in heparinized test tubes, centrifuged at16000r·min^-1for10min, the upper plasma set in theordinary dry test tube, and stored at-80℃. Urine samples were collected at0（pre-dose）, and0-4h,4-8h,8-12h and12-24h post-dose, recording the volume and retention10mL, placedin-80℃. The LC-MS/MS method applied to determination of the concentration ofzaltoprofen in plasma samples and urine samples, and using the WinNonlin6.2thecalculation pharmacokinetic parameters of zaltoprofen.Result: After administration of single80,160or240mg of zaltoprofen, Thepharmacokinetic parameters of zaltprofen: Tmax:1.95±0.56h,2.05±0.50h,2.10±0.46h;C_max:6.63±0.90g·mL^-1,12.48±1.57g·mL^-1,17.82±2.01g·mL^-1; t1/2：4.03±1.37h,4.05±1.38h,4.18±1.61h; AUC_0-t:25.50±7.55g·mL^-1,49.43±5.21g·mL^-1,72.42±12.39g·mL^-1. Urine cumulative excretion rates of0-24h：20.35±2.13%,22.07±3.60%,23.19±4.15%.Conclusion: AUC_0-24and C_maxwere found to be a linear relationship with dose in the rangedose of80²40mg, r>0.9.3. Multi-dose pharmacokinetics study of zaltoprofenObjective: To detect zaltoprofen in plasma and evaluate the multiple pharmacokineticcharacteristics for absorption, distribution and eliminate of zaltoprofen in human.Methods: After single dose administration, the volunteers of low doses group continue to begiven medicine, every time give dosage80mg,3times per day, continuous to be givenmedicine for6days.4ml blood samples were collected according to the time schedule,which included samples just prior to dosing in the morning of the3thto6thdays, and then at0（pre-dose） and0.5,0.75,1.0,1.5,2.0,3.0,4.0,5.0,6.0,8.0,10.0,12.0and24.0h post-dosein the6th day. placed in heparinized test tubes, centrifuged at16000r·min^-1for10min, theupper plasma set in the ordinary dry test tube, and stored at-80℃.Result: After administration of multiple80mg of zaltoprofen, The pharmacokineticparameters of zaltprofen: T_max:1.7±0.59h; C_max:6.45±0.98ng·mL^-1; C_min:0.60±0.19 g·mL^-1; Cav:2.74±0.35g·mL^-1; t1/2:4.13±1.83h； AUC（0-t）:26.34±3.83ng·mL^-1；AUCss:21.94±2.76ng·mL^-1ng·mL^-1. F（Accumulation factor）：0.99±0.32.Conclusion: The multiple pharmacokinetic characteristics is similar to the singlepharmacokinetic characteristics. The Tmax, t1/2and AUC0-thave no significant difference.There is no accumulation of zaltoprofen in body. Under this dose zaltoprofen maybe had afavourable safety.4. Bioequivalence study of zaltoprofenObjective: The aim of this study was to compare the pharmacokinetics and bioequivalenceof zaltoprofen capsule （test） and tablet （reference） formulations.Methods: A single-dose, randomized-sequence, open-label,2-period crossover study wasconducted in healthy Chinese male volunteers. Eligible subjects were randomly assigned at a1:1ratio to receive a single160mg dose of the test or reference formulation, followed by a1-week washout period and administration of the alternate formulation. Venous bloodsamples collecting time is same as singled-dose pharmacokinetics. The blood samplescollecting time schedule is same as single dose pharmacokinetics study.Result: Twenty Chinese male subjects enrolled; all completed the study. No period orsequence effect was observed. The pharmacokinetic parameters of zaltoprofen capsule （test）and tablet （reference）: Tmax:1.90±0.60h,2.20±0.50h: C_max:12.52±2.08g·mL^-1,12.92±2.60g·mL^-1; t1/2:3.92±2.46h,3.96±1.56h; AUC-10-t:55.45±12.20g·h·mL,55.45±11.86g·h·mL^-1; AUC0-∞:56.63±16.66g·h·mL^-1,56.60±13.10g·h·mL^-1. The90%CIsfor the log-transformed ratios of C_max, AUC（0-24）, and AUC0-∞were89.9to106.2,94.3to103.8, and94.5to103.6, respectively （all, P>0.05）. No AEs occurred or were reportedduring the study.Conclusions: In this study of healthy Chinese adult male volunteers, a single160mg dose ofthe zaltoprofen capsule was bioequivalent to a single160mg dose of the tablet formulationbased on the Chinese SFDA’s regulatory definition （rate and extent of absorption）. Both formulations were well tolerated.