JP6124273B2 - 分子イメージングにより代謝機能を測定するための検査薬 - Google Patents
分子イメージングにより代謝機能を測定するための検査薬 Download PDFInfo
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- JP6124273B2 JP6124273B2 JP2012044231A JP2012044231A JP6124273B2 JP 6124273 B2 JP6124273 B2 JP 6124273B2 JP 2012044231 A JP2012044231 A JP 2012044231A JP 2012044231 A JP2012044231 A JP 2012044231A JP 6124273 B2 JP6124273 B2 JP 6124273B2
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Description
(1)生体内に存在する酵素の作用により組織から排出されやすい放射性代謝物、蛍光性代謝物、常磁性代謝物又は磁気共鳴性代謝物に変化する化合物を含有する、分子イメージングにより代謝機能を測定するための検査薬。
(2)前記化合物が、放射性化合物であって、生体内に存在する酵素の作用により組織から排出されやすい放射性代謝物に変化する放射性化合物である前記(1)に記載の検査薬。
(3)前記化合物が、蛍光性化合物であって、生体内に存在する酵素の作用により組織から排出されやすい蛍光性代謝物に変化する蛍光性化合物である前記(1)に記載の検査薬。
(4)前記化合物が、常磁性化合物であって、生体内に存在する酵素の作用により組織から排出されやすい常磁性代謝物に変化する常磁性化合物である前記(1)に記載の検査薬。
(5)前記化合物が、磁気共鳴性化合物であって、生体内に存在する酵素の作用により組織から排出されやすい磁気共鳴性代謝物に変化する磁気共鳴性化合物である前記(1)に記載の検査薬。
(CYP1A2)
amitriptyline, caffeine, clomipramine, clozapine, cyclobenzaprine, estradiol, fluvoxamine, haloperidol, imipramine, mexiletine, naproxen, olanzapine, ondansetron, acetaminophen, propranolol, riluzole, ropivacaine, tacrine, theophylline, tizanidine, verapamil, R-warfarin, zileuton, zolmitriptan
(CYP2B6)
bupropion, cyclophosphamide, efavirenz, ifosphamide, methadone, sorafenib
(CYP2C8)
amodiaquine, cerivastatin, paclitaxel, repaglinide, sorafenib, torsemide
(CYP2C9)
非ステロイド性抗炎症薬:diclofenac, ibuprofen, lornoxicam, meloxicam, S-naproxen, piroxicam, suprofen
経口血糖降下剤:tolbutamide, glipizide
アンジオテンシンII阻害薬:losartan, irbesartan
スルホニル尿素系薬剤:glyburide, glibenclamide, glipizide, glimepiride, tolbutamide
その他:amitriptyline, celecoxib, fluoxetine, fluvastatin, glyburide, nateglinide, phenytoin-4-OH2, rosiglitazone, tamoxifen, torsemide, S-warfarin
(CYP2C19)
プロトンポンプ阻害薬:lansoprazole, omeprazole, pantoprazole, rabeprazole
抗てんかん剤:diazepam, phenytoin, S-mephenytoin, phenobarbitone
その他:amitriptyline, carisoprodol, citalopram, chloramphenicol, clomipramine, clopidogrel, cyclophosphamide, hexobarbital, imipramine, indomethacin, R-mephobarbital, moclobemide, nelfinavir, nilutamide, primidone, progesterone, proguanil, propranolol, teniposide, R-warfarin
(CYP2D6)
tamoxifen
β遮断薬:carvedilol, S-metoprolol, propafenone, timolol
抗うつ薬:amitriptyline, clomipramine, desipramine, fluoxetine, imipramine, paroxetine
抗精神病薬:haloperidol, perphenazine, risperidone, thioridazine, zuclopenthixol
その他:alprenolol, amphetamine, aripiprazole, atomoxetine, bufuralol, chlorphenimipramine, chlorpromazine, codeine, debrisoquine, dexfenfluramine, dextromethorphan, donepezil, duloxetine, encainide, flecainide, fluvoxamine, lidocaine, metoclopramide, methoxyamphetamine, mexiletine, minaprine, nebivolol, nortriptyline, ondansetron, oxycodone, perhexiline, phenacetin, phenformin, promethazine, propranolol, sparteine, tramadol, venlafaxine
(CYP2E1)
麻酔薬:enflurane, halothane, isoflurane, methoxyflurane, sevoflurane
その他:acetaminophen, aniline, benzene, chlorzoxazone, ethanol, N,N-dimethylformamide, theophylline
(CYP3A4,5,7)
マクロライド系抗生物質:clarithromycin, erythromycin (not 3A5)
抗不整脈薬:quinidine (not 3A5)
ベンゾジアゼピン系薬物:alprazolam, diazepam, midazolam, triazolam
免疫調節剤:cyclosporine, tacrolimus (FK506)
抗HIV薬:indinavir, nelfinavir, ritonavir, saquinavir
消化管運動改善薬:cisapride
抗ヒスタミン薬:astemizole, chlorpheniramine, terfenadine
カルシウム拮抗薬:amlodipine, diltiazem, felodipine, lercanidipine, nifedipine, nisoldipine, nitrendipine, verapamil
HMG−CoA還元酵素阻害薬:atorvastatin, cerivastatin, lovastatin, simvastatin
ステロイド類:estradiol, hydrocortisone, progesterone, testosterone
その他:alfentanil, aprepitant, aripiprazole, boceprevir, buspirone, caffeine, cilostazol, cocaine, dapsone, dexamethasone, dextromethorphan, docetaxel, domperidone, eplerenone, fentanyl, finasteride, gleevec, haloperidol, imipramine, irinotecan, LAAM, lidocaine, methadone, nateglinide, ondansetron, pimozide, propranolol, quetiapine, quinine, risperidone, salmeterol, sildenafil, sirolimus, sorafenib, sunitinib, tamoxifen, taxol, telaprevir, terfenadine, torisel, trazodone, vincristine, zaleplon, ziprasidone, zolpidem
本発明においては、目的に応じて、前記放射性化合物、蛍光性化合物、常磁性化合物又は磁気共鳴性化合物を単独で、又は2種以上組み合わせて用いる。
コラゲナーゼ還流法により調製したラット遊離肝細胞を2×106個/mLになるように蒸留水に懸濁し、ホモジナイザーで機械的にホモジネートを調製した。このホモジネート200μLに対し、蒸留水で20μMの濃度に調製した5 (and 6)-carboxy-2’,7’-dichlorofluorescein diacetate(CDFDA)を加え(終濃度10μM)、37℃で5分間インキュベーションした。400μLの氷冷1%Tritonを加えて反応を停止させ、高速遠心操作(15000rpm、10min、4℃)により上清に含まれる5 (and 6)-carboxy-2’,7’-dichlorofluorescein(CDF)及びCDFDAを回収した。CDFの蛍光強度は蛍光プレートリーダーで測定した。99℃で5分間熱変性させたホモジネートではCDFの蛍光がほぼ観察されず、その蛍光強度は熱変性しなかった場合の約2%程度であった(図3)。このことからCDFDAがラット肝細胞中に含まれる酵素により代謝され、CDFを含む蛍光物質が得られることがわかった。
文献(Dunn, J., Yarmush, M., Koebe, H., & Tompkins, R.: Hepatocyte function and extracellular matrix geometry: long-term culture in a sandwich configuration. FASEB J., 3: 174-177, 1989)記載の方法に従い、ラット肝から遊離肝細胞を単離後、予めコラーゲンコートしておいた35mmプレートに播種した。細胞を接着させるために37℃、5%CO2条件下で2〜6時間培養した後、William’s E medium(WEM)へと培地を交換した。播種から24時間後に培地をマトリゲル含有WEMへ交換し、細胞にマトリゲルを重層しサンドイッチ培養した。播種後4日後に実験に供した。敷石上に培養された細胞間隙にサンドイッチ培養ラット肝細胞(SCRH)特徴的な胆管腔の形成が促された。次に、細胞内で蛍光物質に代謝されるdihydrofluorescein diacetate(H2FDA)10μMをサンドイッチ培養したラット肝細胞とインキュベートし(37℃、pH7.4)、蛍光顕微鏡により胆管腔に蓄積される蛍光物質の蛍光強度を経時的に測定し、定量化した。H2FDAと胆汁酸トランスポーターBSEP(Bile Salt Efflux Pump)の阻害剤であるトログリタゾン50μMを共存させた場合(図4、○)、非存在時(図4、●)で有意な蛍光が観察された6分以降15分までの蛍光強度は非存在時(図4、●)と比較して有意に減少した。
ラット肝細胞内への蓄積を増加させるために脂溶性を高め、未代謝状態でも蛍光を持つように合成された加水分解酵素で構造変換する蛍光性化合物30μMをサンドイッチ培養したラット肝細胞に取込ませ、蛍光顕微鏡により胆管腔に蓄積される蛍光物質の蛍光強度を測定し、定量化した。前記蛍光性化合物と胆汁酸トランスポーターBSEPの阻害剤であるトログリタゾン50μMを共存させた場合(図5、○)、24分までの蛍光強度は非存在時(図5、●)と比較して有意に減少した。
放射性核種標識アミノ酸として、L−又はD−メチオニン(L−/D−Met)の[S−methyl−3H]標識体(S−L−/D−Met;American Radiolabeled Chemicals)と[1−carboxyl−14C]標識体(1−L−/D−Met;American Radiolabeled Chemicals)を用い、ダブルトレーサー法により検討した。
125I標識イオマゼニル(125I−IMZ(臨床PET製剤は123I標識体))は日本メジフィジックス社より提供を受けた。
Claims (2)
- 15−(4−ヨードフェニル)−3(R、S)−メチルペンタデカン酸( 123 I)、 11 C−パルミチン酸及び 123 I−ヘプタデカン酸のC 1−6 アルキルエステル、並びに 125 I標識イオマゼニルから選ばれるエステル化合物を含有する、分子イメージングにより、肝エステラーゼによるエステル基の加水分解機能を測定するための検査薬。
- 前記エステル化合物が125I標識イオマゼニルである請求項1記載の検査薬。
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