보고서 정보
| 주관연구기관 |
인제대학교
Inje University |
| 연구책임자 |
김동현
|
| 보고서유형 | 최종보고서 |
|---|
| 발행국가 | 대한민국 |
|---|
| 언어 |
한국어
|
| 발행년월 | 2015-08 |
|---|
| 주관부처 |
보건복지부
[Ministry of Health & Welfare(MW)(MW) |
| 연구관리전문기관 |
질병관리본부
Korea Center for Diease Control and Prevention |
| 등록번호 |
TRKO201900001092 |
| DB 구축일자 |
2019-06-29
|
| 키워드 |
항결핵제.다제내성.치료적 약물 모니터링.Anti-tuberculosis agent.multi-resistance.LC/MS/MS.TDM.
|
초록
▼
1,2차 결핵 환자에서 항결핵제의 한국인 고유의 약동학 파라미터를 얻고, 치료적 약물 모니터링 방법을 확립하고자 하였다. 먼저 인체 혈장에서 20종의 항결핵제를 동시분석하는 LC/MS/MS 방법을 확립하였다. 시료 전처리는 group 1 (amikacin, kanamycin, streptomycin and pyrazinamide)은 0.13N HCl을 이용하여, 그리고 group 2 (amoxicillin, ciprofloxacin, clarithromycin, clofazimine, cycloserine, ethambutol, e
1,2차 결핵 환자에서 항결핵제의 한국인 고유의 약동학 파라미터를 얻고, 치료적 약물 모니터링 방법을 확립하고자 하였다. 먼저 인체 혈장에서 20종의 항결핵제를 동시분석하는 LC/MS/MS 방법을 확립하였다. 시료 전처리는 group 1 (amikacin, kanamycin, streptomycin and pyrazinamide)은 0.13N HCl을 이용하여, 그리고 group 2 (amoxicillin, ciprofloxacin, clarithromycin, clofazimine, cycloserine, ethambutol, ethionamide, isoniazid, levofloxacin, linezolid, moxifloxacin, p-aminosalicylic acid(PAS), prothionamide, rifabutin, rifampin and roxithromycin)는 acetonitrile을 이용하여 제단백하였다. 분리는 Hilic Silica column (150 mm × 2.1 mm)과 Atlantis C18 column (50mm×2.0mm i.d., 3μm)을 이용하여 gradient 방법을 적용하였고 검출은 MRM방법을 이용하였다. 확립된 방법은 정량 범위에서 우수한 직선성 (r > 0.9969)을 보여 주었고 일내, 그리고 일간 분석오차는 14.3%이내, 그리고 정확성은 84.8에서 113%로 우수하였다. 한국인 결핵환자 고유의 약동학 파라미터는 1차 결핵환자48명, 2차 결핵 환자 30명을 대상으로 집단 약동학 방법을 이용하여 분석하였다. 1차 약제 4종 isoniazid, rifampin, pyrazinamide, 그리고 ethambutol의 Cl, Vd, 그리고 ka값을 얻어 타인종의 파라미터와 비교 시 차이를 보여 주었으며, 이러한 파라미터를 이용하여 예측된 개개 약물 농도와 실측치를 비교 분석한 결과 본 시험에서 얻은 파라미터가 한국인 환자의 약물 농도 에측에 더 부합함을 입증하였다. 30명의 다제내성 환자를 대상으로 한 2차 약제의 약동학 파라미터는 먼저 분석에 충분한 시료수가 확보된 cycloserine, moxifloxacin, prothioamide, 그리고 amoxicilin에 대한 약동학 파라미터를 분석하였다. Cycloserine의 경우 Cl, Vd, 그리고 ka값이 각각 0.912 L/hr, 24.2 L, 그리고 0.788 hr- 로서 계산되었고 이 값을 얻은 모형의 유의성을 visual predictive check등 여러 방법을 사용하여 검증하였다. 또한 나머지 3종 약물도 동일한 방법을 이용하여 약동학 파라미터를 얻고 검증을 하였다. 또한 치료 성공률이 낮은 다제내성 환자의 약물 농도와 음전율의 상관성을 분석한 결과 비음전군에서 moxifloxacin의 약물 농도가 상대적으로 낮은 것으로 판명되었다. 또한 prothioamide의 경우 비음전군에서 약물 농도가 teraget range에 미치지 못하는 환자수가 음전군에 비하여 상대적으로 많은 것으로 나타났다. 제한적인 환자 수에 의하여 통계적 유의성은 입증하지 못하였으나, 약물 농도가 치료 성공률에 영향을 미치는 것을 시사하며, 이는 치료적 약물 농도 모니터링을 통하여 용량 변경이 필요함을 시사한다, 다 빈도로 사용되는 2차 약제의 약동학 파라미터도 추가 시험을 통하여 분석하고 이를 기반으로 하리라 사료된다.
(출처 : 보고서 요약문 4p)
Abstract
▼
An accurate liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method for the quantitation of 20 anti-tuberculosis (anti-TB) drugs in human plasma, was developed as a tool for therapeutic drug monitoring. Two protein precipitation methods were adopted; one using methanol containing 0.13 N H
An accurate liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method for the quantitation of 20 anti-tuberculosis (anti-TB) drugs in human plasma, was developed as a tool for therapeutic drug monitoring. Two protein precipitation methods were adopted; one using methanol containing 0.13 N HCl, for precipitation of amikacin, kanamycin, streptomycin and pyrazinamide, and the other using acetonitrile, for precipitation of preamoxicillin, ciprofloxacin, clarithromycin, clofazimine, cycloserine, ethambutol, ethionamide, isoniazid, levofloxacin, linezolid, moxifloxacin, p-aminosalicylic acid (PAS), prothionamide, rifabutin, rifampin and roxithromycin. Separation was performed either on an HILIC silica column or a reversed-phase dC18 column, with a gradient elution. Detection was carried out in multiple reaction-monitoring (MRM) mode. The calibration curves were linear over a 50-fold concentration range, with correlation coefficients (r) greater than 0.9969 for all anti-TB drugs. The intra- and inter-day precision was less than 14.3%, and the accuracy ranged between 84.8 and 113.0%. Pharmacokinetic parameters of 1st, and 2nd anti-TB drugs in Korean patients were obtained from 48 TB patients and 30 MDR TB patients by using population pharmacokinetics. Cl, Vd, and ka values for isoniazid, rifampin, pyrazinamide, and ethambutol obtained in this study showed small differences compared with those in other population published elsewhere. The predicted plasma concentration using these parameters were highly correlated with actual plasma concentrations, suggesting that PK parameters calculated from Korean patients should be applied to acual TDM. PK parameters of 2nd line anti-TB drugs form MDR TB patients were initially estimated for cycloserine, moxifloxacin, prothioamide, and amoxicilin due to number of plasma samples. In case of cycloserine, Cl, Vd, and ka values were calculated to be 0.912 L/hr, 24.2 L, and 0.788 hr- respectively. The population pharmacokinetic model for cycloserine was validated by several methods including visual predictive check. The popuplation PK parameters for the rest of drugs were also calculated and validated using the same approach. Plsam concentrations of the drugs were carefully compared with 4- and 6 month conversion rates to check whether theraputic failure in MDR-TB patients is related with low drug concentrations. Low plasma concentration sof moxifloxacin seemed to be associated with therapeutic failure. In addition, the percent of plasma concentrations of prothioamide not reached to target therapeutic concentration range was higher in non-conversion group. Although no statistically significant difference was observed in these analysis due to limited patient numbers, thses results justified the importance of dose adjustment for the treatment of MDR-TB patient by therapeutic drug monitoring in order tomaintain proper plasma drug concentrations. In addition, PK parameters of drugs frequently prescribed in MDR-TB except for the drugs analyzed in this study should be further evaluated in th future to enhance therapeutic success in the treatment of MDR-TB.
(출처 : Summary 5p)
목차 Contents
- 표지 ... 1
- 정책연구용역사업 최종결과보고서 ... 2
- 목차 ... 3
- 보고서 요약문 ... 4
- Summary ... 5
- 정책연구용역사업 총괄 연구결과 ... 6
- 제1장 최종 목표 ... 6
- 제2장 국내외 기술 현황 ... 8
- 제3장 최종 연구내용 및 방법 ... 13
- 제4장 최종 연구결과 ... 26
- 제5장 연구결과 고찰 및 결론 ... 111
- 제6장 연구성과 및 활용계획 ... 114
- 제7장 정책연구용역사업 진행과정에서 수집한 해외과학기술정보 ... 116
- 제8장 기타 중요변경사항 ... 116
- 제9장 연구비 사용 내역 및 연구원 분담표 ... 117
- 제10장 참고문헌 ... 118
- 제11장 첨부서류 ... 121
- 제1세부 연구결과 ... 136
- 제1장 제1세부 최종 목표 ... 136
- 제2장 제1세부 최종 연구내용 및 방법 ... 136
- 제3장 제1세부 최종 연구결과 ... 148
- 제4장 연구결과 고찰 및 결론 ... 224
- 제5장 연구성과 및 활용계획 ... 226
- 제6장 연구비 사용 내역 및 연구원 분담표 ... 228
- 제7장 참고문헌 ... 229
- 제2세부 연구결과 ... 230
- 제1장 제2세부 당해연도 연구목표 ... 230
- 제2장 제2세부 당해연도 연구내용 및 방법 ... 230
- 제3장 제2세부 당해연도 연구결과 ... 232
- 제4장 연구결과 고찰 및 결론 ... 237
- 제5장 제2세부 정책연구용역사업의 연구성과 ... 239
- 제6장 제2세부 연구비 집행 실적 ... 240
- 제7장 참고문헌 ... 240
- 제8장 첨부서류 ... 241
- 끝페이지 ... 241
![The effect of tuberculosis agents on accumulation of [3H]Methotrexate in MDCK-MRP2 cells. [3H]Methotrexate was added and allowed to incubate for 30min at 37℃. Each bar represents mean ± SD of accumulation from three replicate experiments](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_106_image_214.png "The effect of tuberculosis agents on accumulation of [3H]Methotrexate in MDCK-MRP2 cells. [3H]Methotrexate was added and allowed to incubate for 30min at 37℃. Each bar represents mean ± SD of accumulation from three replicate experiments")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]Estrone-3-sulfate into X. laevis oocytes expressing OAT1B1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_107_image_1.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]Estrone-3-sulfate into X. laevis oocytes expressing OAT1B1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]MPP+ into X. laevis oocytes expressing OCT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_108_image_1.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]MPP+ into X. laevis oocytes expressing OCT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [14C]PAH into X. laevis oocytes expressing OAT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_108_image_2.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [14C]PAH into X. laevis oocytes expressing OAT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
![The [14C]PAH uptake with various concentrations (1,10, 50, 100, 200 , 300 and 400 μM) of linezolid was measured for 60 min. Data represent the mean ± S.E. of 8 - 10 independent experiments](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_110_image_217.png "The [14C]PAH uptake with various concentrations (1,10, 50, 100, 200 , 300 and 400 μM) of linezolid was measured for 60 min. Data represent the mean ± S.E. of 8 - 10 independent experiments")
![The effect of tuberculosis agents on accumulation of [3H]Methotrexate in MDCK-MRP2 cells. [3H]Methotrexate was added and allowed to incubate for 30min at 37℃. Each bar represents mean ± SD of accumulation from three replicate experiments](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_220_image_1.png "The effect of tuberculosis agents on accumulation of [3H]Methotrexate in MDCK-MRP2 cells. [3H]Methotrexate was added and allowed to incubate for 30min at 37℃. Each bar represents mean ± SD of accumulation from three replicate experiments")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]Estrone-3-sulfate into X. laevis oocytes expressing OAT1B1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_221_image_1.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]Estrone-3-sulfate into X. laevis oocytes expressing OAT1B1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]MPP+ into X. laevis oocytes expressing OCT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_222_image_1.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [3H]MPP+ into X. laevis oocytes expressing OCT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
![Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [14C]PAH into X. laevis oocytes expressing OAT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group](https://nrms.kisti.re.kr/bitextimages/TRKO201900001092/TRKO201900001092_222_image_215.png "Inhibitory effects of tuberculosis agents (100 μM) on the uptake of [14C]PAH into X. laevis oocytes expressing OAT1 transporter, respectively, was measured for 60 min. Each data point represents the mean ± S.E. of at least 8 oocytes per transport group")
※ AI-Helper는 부적절한 답변을 할 수 있습니다.