Original
©2012 Dustri-Verlag Dr. K. Feistle ISSN 0946-1965
DOI 10.5414/CP201661e-pub: nnn
*Sponsor
Received
发展的意思
September 13, 2011;
accepted
March 13, 2012
Correspondence to
分段函数定义域Prof. Shengjun Zhan,
MD, MBA, CCRP
Zhengzhou University, 1
Eastern Jianshe Road,
Zhengzhou, Henen
450052 China
Key words
paroxetine – pilot – piv-
otal – bioequivalence –
pharmacokinetics – LC/
MS/MS
A pilot study for a pivotal bioequivalence trial using two paroxetine 40 mg tablet formulations in healthy Chine subjects Shengjun Zhang 1, Quancheng Kan *1, Jianguo Wen 1, Jie Zhao 1, Yuqiao Sheng 1, Yidong Li 1, Suke Sun 1, Feng Zhang 1, Li Yang 1, Wenzhe Lv 2, Yuanchao (Derek)
Zhang 2, Kun Qian 2 and Zhongping John Lin*21The First Affiliated Hospital of Zhengzhou University, Henan Province, Zhengzhou, Henan, China and 2Frontage Laboratories, Inc., Exton, PA, USA Abstract. The purpo of this study was to conduct a pilot study in order to ob-tain reliable results for further planning of a well-designed pivotal trial comparing the bioequivalence (BE) of two paroxetine tab-let formulations in healthy Chine subjects. Before conducting the pivotal trial, the pilot trial enrolled 14 subjects to help in study design, establishing the recruitment period, determining pharmacokinetics (PK) time points and sample size, and asssing BE of the two formulations. The single-center, randomized, open-label, single-do, two-period crossover study with a 7-day wash-out interval was conducted after obtaining information from the fasted pilot trial in 72 healthy volunteers for a pivotal study under fed and fasted conditions, respectively. There were 19 PK sample collection time points employed in both the pilot and pivotal trials. A nsitive and specific liquid chromatog -raphy-tandem mass spectrometry (LC-MS/MS) method was developed and validated for determining paroxetine in human plasma. BE between two articles was determined by calculating 90% confidence intervals (CIs) for the ratio of C max 91.38 – 110.39% for the pilot trial, 99.81 – 114.08% for pivotal trial under fasted condition, and 94.06 – 110.41% for pivotal trial under fed condition, AUC (0–t) 96.06 – 110.52% for pilot study, 100.88 – 113.05% for the pivotal trial un-der fasted condition, and 97.08 – 106.06% for pivotal study under fed condition, and AUC (0–∞) 9
6.17 – 110.42% for the pilot study, 100.85 – 112.81% for the pivotal trial under fasted condition and 97.22 – 106.14% for the pivotal study under fed condition, respectively. The values for the test and reference products are within the 80 – 125% interval propod by FDA and EMEA. It was concluded that the propod method was successfully applied to a PK study in healthy Chine volunteers, and results showed from both the pilot and pivotal studies that the two paroxetine formulations are bioequivalent in their rates and extent of absorption.Introduction Major depressive disorder (MDD) is a common, chronic illness associated with substantial disability and economic burden. The National Comorbidity Survey Replica-tion (NCS-R) study recently estimated the lifetime prevalence of MDD in the United States to be 16% (95% CI, 15.1 – 17.3%) and the 12-month prevalence to be 6.6% (95% CI, 5.9 – 7.3%) [1]. A study from Da -vid et al. [2], found that 32.6 – 35.1 million adults in the United States were projected to experience MDD during their lifetime, with 13.1 – 14.2 million affected during the 12 months before the survey. Although a num-ber of effective antidepressants are available, respon to individual antidepressant agents is variable. About 30 – 45% of patients with MDD respond only partially or not at all to antidepressants, owing to lack of efficacy or intolerance [3]. Therefore, there remains an unmet medical need for new medications that are both effective and well tolerated.Paroxetine (Figure 1) is a lective ro-tonin reuptake inhibitor (SSRI) antidepres -sant. The chemical formula is C 19H 20FNO 3. It is ud to treat major depression, obssive-comp
ulsive, panic, social anxiety, and gener-alized anxiety disorders in adult outpatients. A meta-analysis of the databa of worldwide clinical trials of paroxetine found its efficacy to be equivalent to that of tricyclic antidepres-sants (TCAs) in patients with melancholia and vere MDD [4]. Moreover, a study in elderly depresd patients reported paroxetine to be equivalent in efficacy to nortriptyline, with a more favorable tolerability profile [5].There are two steps, which are usually followed to determine bioequivalence (BE).
International Journal of Clinical Pharmacology and Therapeutics, Vol. nn – No. nn /2012 (1-10)
Zhang, Kan, Wen et al. 2
In the first step, a pilot trial is conducted to evaluate the acceptability of the test formula-tion as a candidate for further evaluation in a subquent pivotal BE trial, and to decide if a generic drug shows promi of BE [6]. In the cond step, results from the pilot study are examined to determine the value of proceed-ing with a pivotal study. Information col -lected from a pilot study can be very uful in determining PK sample size, the design of the following pivotal trial, and other vari-ables. Therefore, a pilot trial was conducted in this paroxetine study before carrying out the pivotal BE trial.The test formulation being evaluated in the study is a new generic formulation of paroxetine 40 mg tablets developed by Hua-hai Pharmaceutical Co., Ltd. The BE of the test formulation is reported here for the first time. In the trial, healthy Chine male and female volunteers were enrolled to investi-gate the BE between the reference (R) for-mulation PAXIL ® paroxetine 40 mg tablets (GlaxoSmithKline, Batch No. 8H004), and the test (T) formulation Paroxetine 40 mg tablets (Huaihai Pharmaceutical Co., Ltd., Batch No. 971309001A).The study objective of this pilot trail was to obtain reliable results for further planning of a well-designed pivotal study comparing the BE of two paroxetine tablet formulations in healthy Chine subjects.Subjects and methods Subjects The study was open to healthy Chine adult males and females between 18 and 45
years old, who are at least 60 kg (132 lbs) for men and 48 kg (106 lbs) for women with a body mass i
ndex (BMI) between 19 and 30, and a negative pregnancy test result if female. Potential subjects were excluded if they were current smokers or urs of any tobacco products, and/or had histories of hypernsitivity to paroxetine or any other component of the paroxetine tablets, and/or had ud investigational drugs or prod-ucts, or participated in a drug rearch study within a period of 30 days prior to receiving the investigational drug, and/or ud any pre-scription drug therapy within 14 days prior to receiving the investigational drug. They were also excluded if they had donated blood (1 pint or more) within 30 days or plasma within 7 days of receiving the investigational drug, and/or had a recent history of alcohol-ism (< 2 years) or ud alcohol within 24 h prior to receiving investigational drug. Fi-nally, they were excluded if they had any clinically significant abnormality bad on medical history, physical examination, and laboratory analysis.
The study protocols and informed con-nt forms were approved by Institutional Review Board (IRB) (IRB00007169) of Zhengzhou University First Affiliated Hos -pital. Before enrollment, all subjects signed an IRB approved Informed Connt Form and underwent clinical screening, including a physical examination and medical labora-tory tests.
Bad on inclusion and exclusion criteria of the protocol, 14 healthy volunteers were enrolled in a fasted pilot trial. After collect-ing results from the pilot trial, 36 subjects were determined to be eligible
for a fed piv-otal trial. Upon establishing BE of test and reference paroxetine formulations from the fed pivotal trial, 36 subjects were recruited for a fasted pivotal trial.Study design and procedures
The studies were single-center, random-ized, open-label, single-do, two-period, with crossover design to asss the BE of ref-erence (R) and test (T) formulations of par-oxetine 40 mg oral tablets in healthy Chine adult female and male subjects. After singed
informed connt forms were obtained, vol-Figure 1. Chemical structure of paroxetine ( en.wikipedia/wiki/Paroxetine).
Pilot study for a bioequivalence trial of two paroxetine 40 mg tablet formulations 3
unteers underwent a complete medical histo-ry, physical examination, medication history, vital sign evaluation (sitting blood pressure, pul rate, and temperature), resting ECG, clinical laboratory tests (chemistry, hematol-ogy, urinalysis, hepatitis B & C diagnostic profile), and a urine pregnancy test for fe-males only.
14, 36/36 healthy volunteers, who fit the inclusion and exclusion criteria of the protocol, were enrolled in fasted pilot trial, and fed/fasted pivotal trial, respectively. Randomly, subjects were divided
into two groups, and assigned with (T-R) or (R-T) treatment quence according to the random-ization schedule, which was prepared before admission to the study center.
Subjects reported to the rearch facility at ~ 20:00 hours on Day –1, and remained confined to the rearch facility until ~ 48 h after the study medication do. An alcohol test (all subjects) and a urine pregnancy test (females only) had been performed upon ad-mission to the rearch facility.
On Session 1, Day 1, following an over-night fast of at least 10 h, subjects consumed a high fat breakfast ingested 30 min prior to dosing for the fed pivotal study. For the fasted pilot and pivotal studies, subjects did not con-sume breakfast on the dosing day. Subjects received a single oral do of reference or test formulation of paroxetine 40 mg tablets with 240 ml of tepid water and remained sitting up-right with ambulation limited during the 2-h post-do period. Subjects remained confined to the rearch facility until 48 h post do. Following a washout period of 7 days, sub-jects were crosd over to alternate the dosing of the test and reference formulations on Ses-sion 2 Day 22. The same PK time points were procesd as in Session 1.
Serial blood samples for PK analysis were obtained at Time 0 (within 30 min pre-do) 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 24, 36, 48, 72, 96, 120, 144 and 168 h post-do. Blood samples were collected into
chilled blood collection tubes containing K2EDTA. After collection the samples were imme-diately centrifuged at 2,000 g for 10 min at 4 ºC. Plasma was parated from collection tubes, and samples were stored on dry ice (~ –70 °C) until shipped to Frontage Labora-tories (Shanghai), Inc. (Shanghai, China) for LC/MS/MS analysis. The whole procedure from blood collecting to plasma storage was carried out in less than 60 min.
Adver event and rious adver events (AE/SAE) were monitored throughout the study bad on direct questioning, spontane-ous reports, and clinical parameters (blood pressure, pul and temperature) with mea-surement at Time 0 (within 30 min pre-do), 1, 4, 8, 24, 48, 72, 96, 120, 144 and 168 h post do. Following collection during Ses-sion 2, and before discharged from the clini-cal rearch center, an abbreviated physical examination was performed on all subjects. Blood and urine samples were collected for clinical safety laboratory tests (chemistry, hematology and urinalysis). Any AE (clini-cal sign, symptom, or dia) temporally as-sociated with the u of this investigational drug was documented, whether or not they were considered related to the investigation-al drug. All AEs occurring during this study, including during the washout interval, were documented.xxxx影院
The high fat breakfast meal contained ~ 150 protein calories, 250 carbohydrate calories, and 500 – 6
00 fat calories. The breakfast distributed to all fed study subjects included 2 eggs fried in oil, 2 strips of pork bacon, 2 slices of toast with butter, 4 ounc-es of hash brown potatoes and 8 ounces of whole milk.
The studies were performed in accor-dance with the ethical standards for studies in humans stipulated by the Declaration of Helsinki and its amendments, the Interna-tional Conference on Harmonization Guide-line for Good Clinical Practice (ICH), and the Guideline for Good Clinical Practice recommended by the guidelines of the U.S. Food and Drug Administration (FDA) and China’s State Food and Drug Administration (SFDA) [7, 8, 9, 10].
广州机动车LC-MS/MS Method
Paroxetine concentrations in plasma were measured using a validated liquid chromatography-tandem mass spectrom-etry (LC/MS/MS) method. Plasma samples were extracted using liquid-liquid extraction. Plasma (200 µl) was mixed with 20 µl of di-luent (methanol/water 50 : 50) for double blank, 20 µl of Internal Standard Spiking
Zhang, Kan, Wen et al. 4
Solution (paroxetine-d6, 250 ng/ml) for other samples, 200 µl of 0.1M NaOH solution and 2 ml of methyl t-butyl ether as the extract-ing solvent. The samples were vortexed for 10 min followed by centrifugation at 3,500 rpm for 5 min. The upper organic layers were transferred to clean test tubes and evaporated to dryness at 40 °C under gentle nitrogen flow. The extracted samples were reconsti-tuted with 150 µl of reconstitution solution (MeOH/water 25/75) and a 10-µl sample was injected to the LC/MS/MS system – Sci-ex API 4000 coupled to Shimadzu LC pump and autosampler. Chromatographic para-tion was achieved by using Column (Xbridge C18, 30 × 4.6 mm, 3.5 micron) with the mo-bile pha consisting of acetonitrile/water with 0.05% formic acid. The mobile pha was delivered into the LC/MS/MS system at a flow rate of 0.8 ml/min. MS detection was carried out by Multiple Reaction Monitoring (MRM) of m/z 330.2 > 192.2 for paroxetine and 336.2 > 198.2 for paroxetine-d6 (IS), re-spectively. Data acquisition and processing were powered by the Analyst 1.4.2 software package (Applied Biosystems, Foster City, CA, USA).
Pharmacokinetic analysis
A non-compartmental PK method was employed to determine the PK parameters (AUC0–t, AUC0–∞, C max, t max, K el, and t½1/2) of paroxetine by WinNonlin (version 5.0.1, Pharsight Corporation, Mountain View, CA, USA). C max, the maximum obrved concen-tration, and t max, the time to obs
erve the peak concentration, were determined for each sub-ject and for each treatment. The area under the concentration-time curve from Time 0 to the last quantifiable concentration (AUC0–t) was determined by trapezoidal rule. AUC0–∞, the area under the concentration-time curve from Time 0 to infinity, was determined by the trapezoidal rule and extrapolated to in-finity as estimated by the last quantifiable concentration divided by the elimination rate constant (K el). K el was determined by simple linear regression bad on the terminal pha of plasma concentration. Plasma half-life (t1/2) was estimated by (0.693/ K el). In ad-dition to the descriptive statistics, geometric means were reported for the pivotal PK end-points (AUC0–t, AUC0–∞, and C max).
Statistical analysis
Analysis of Variance (ANOV A), 90% confidence intervals (CIs), power analysis and two one-sided tests were ud to make statistical evaluation of PK data and the as-ssment of BE. BE of the test and refer-ence formulations was determined bad on AUC0–t, AUC0–∞ and C max of paroxetine in plasma. The BE was considered if the 90% CIs on the ratio of test (T) to reference (R) formulations were within a range of 80 – 125%. Log-transformed PK parameters AUC0–t, AUC0–∞, and C max were analyzed using analysis of variance (ANOV A) mod-el including terms for quence, formula-tion, and period as fixed effects, and subject nested within quence as a random effect. Sequence
was tested using subject nested within quence as the error term. A CI on the ratio of untransformed PK parameters was derived through rever transformation of the 90% CI for the difference in the log scale to the 90% CI for the ratio in the origi-nal scale.
Adver events/rious adver events (AE/SAE)
Subjects have been monitored through-out confinement for any adver events. The investigators have reviewed each event to asss its potential inducement by the study drug. Each sign or symptom was graded for verity. Duration, resolution, and the date and time of ont, have been recorded in the Ca Report Form (CRF).
All rious event reporting adhered to 21 CFR 312.32 for IND drugs and 21 CFR 314.80 for marketed drugs. If any of the ad-ver events are rious, special procedures would be carried out. All rious adver events have been immediately (within 24 h) reported to the sponsor by telephone and fol-lowed by a written report within 5 working days. All adver events, including rious events, have been followed through to reso-lution regardless of whether the subject was still participating in the study. All adver
虚心好学Pilot study for a bioequivalence trial of two paroxetine 40 mg tablet formulations 5
events and administered treatments have been recorded in the CRF.
Results
LC-MS/MS method validation The method was validated for accuracy, precision, nsitivity, specificity, linearity, and reproducibility according to the FDA guidance for bioanalytical method validation [11] over a concentration range of 0.10 – 100 ng/ml using eight calibration standards and six replicates of QC samples at each con-centration level in three parate batch runs. Each batch run also contained additional samples such as stability samples for pro-cessing and storage.
Analyte stability was tested using QC samples for multiple freeze/thaw cycles (F/T cycles), on the bench at room temperature (short-term stability), or frozen at –70 °C (long-term storage). Post-preparative stabil-ity and stock solution stability were also de-termined. The extraction recovery of parox-etine was calculated by comparing the peak areas of extracted plasma standards to the peak areas of post-extraction plasma blanks spiked at corresponding concentrations. The method specificity was evaluated by screen-ing six lots of blank K2EDTA plasma. The precision and accuracy data for QCs are summarized in Table 1. For QCs at 0.3 ng/ ml (Low QC) and 75 ng/ml (high QC), inter-assay CV values were 5.0% and 2.7%, re-spectively, and the %Nominal ranged fro
m 92.7% to 99.7%. Intra-assay CV values for LLOQ at 0.1 ng/ml were 4.8%, and the %Nominal ranged from 94.0% to 105.0%. The CV and %Nominal values indicated reproducible LC/MS/MS conditions and that the assay is consistent and reliable. For dilution integrity evaluation, QC samples (500 ng/ml) were diluted 20-fold with blank plasma prior to extraction. The dilution in-tegrity data showed a CV of 2.3% with a %Nominal of 94.9%, which supports sample dilution up to 20-fold for analysis. The repre-ntative chromatograms of paroxetine (left) and paroxetine-d6, IS (right) in blank plasma (A), plasma spiked with 5 ng/ml paroxetine (mid QC) and 250 ng/ml paroxetine-d6 (B), and plasma sample equivalent to 28.8 ng/ml from Subject 202, Period, 10 h post-do (C) are prented in Figure 2.机电设备安装
Demographic data
In the pilot study, 14 healthy qualified subjects were enrolled in and completed the study. In the pivotal trial, 72 healthy qualified subjects were enrolled in the study. However, 4 subjects voluntarily withdrew, resulting in 68 subjects who completed the pivotal trial. The demographic characteristics of the study population are summarized in Table 2.
Pharmacokinetic analysis
天资聪慧
手掌心发黄Linear and mi-log plots of mean par-oxetine concentration-time profiles after ad-ministration of a single 40-mg oral do of test and reference formulations to healthy
