Ma muli o ka emi ʻana o nā hana enzyme metabolizing i ka lāʻau me ka hoʻolōʻihi ʻia o ka manawa incubation in vitro, ua palena nā ʻano kuʻuna in vitro metabolism e like me ka manawa incubation pōkole, haʻahaʻa haʻahaʻa haʻahaʻa haʻahaʻa a me ka palena haʻahaʻa haʻahaʻa o ka ʻike (ʻo ka palena haʻahaʻa haʻahaʻa o CLint assay he 2.5 μL / (min · 106 mau cell)), ʻaʻole hiki ke wānana pololei i ka CLint o nā lāʻau lapaʻau metabolized.[1]. I kēia manawa,hepatocytes he nui nā enzymes e pono ai no ka hoʻoemi ʻana, ka hoʻemi ʻana, ka hydrolysis a me nā hopena dimerization i hoʻohālikelike ʻia menā microsomes ate, a i loko o nā ʻōnaehana in vitro, nui nā hiʻohiʻona hou e hoʻomaʻamaʻa ʻia ma muli o kahepatocyte ʻōnaehana a hoʻohālike hou i ke kaiapuni in vivo e hoʻolōʻihi i ka paʻa ʻana o ka hana enzyme i lokohepatocytes, a me ka hoʻolōʻihi ʻia o ka manawa incubation e hāʻawi i ka hiki ke nānā i ka hoʻololi metabolic kūpono o nā lāʻau metabolizing lohi.[1,2].
Ma muli o kēia, ua alakaʻi ʻo IPHASE, ma ke ʻano he alakaʻi i nā reagents biological no ka noiʻi in vitro, ua hoʻokumu i ka HepatoMax ™ System a me HepatoCon ™ Media System e pili ana i ke aʻo ʻana i nā hepatocytes kumu mua a me nā cell stromal o nā ʻano genera like ʻole, me ke kanaka, ka monkey, canine, rat, a me ka ʻiole, a pēlā aku.hepatocytes ma keʻano he laʻana, a ua hoʻonohonoho ʻia nā ʻōnaehana ʻelua no ka hoʻoholo ʻana i ka hana a me ke kūpaʻa metabolic o nā ʻano enzyme kikoʻī a me ka hoʻonui ʻana i ka waiwai pane o nā metabolites o ka lāʻau tolbutamide i nā lā 10 a me 7 mau lā e hōʻoia i nā ʻōnaehana. Hoʻonui ka Metabolite i ka waiwai pane i nā lā 10 a me ka hana a me ka metabolic stability o nā ʻano enzyme kikoʻī i nā lā 7 i hoʻoholo ʻia e hōʻoia i nā ʻōnaehana.
- (1) ʻO nā kaʻina hana moʻomeheu hōʻoia lāʻau ʻelua e like me kēia:
IPHASE-HepatoMax™He ʻōnaehana moʻomeheu like ia o nā hepatocytes kanaka mua a me nā pūnaewele stromal. Ua ho'ōla hou ʻia nā hepatocytes kanaka mua a hoʻoulu ʻia me nā stromal cell i ka HepatoP™ medium no 3 mau lā e hoʻomaka ai i ka hoʻokele lāʻau hoʻomaʻamaʻa ʻana, a ua lawe ʻia ka metabolism o ka lāʻau i ka serum-free HepatoDo™ medium, i hoʻopiha ʻia me ka lāʻau lapaʻau me ka hoʻololi ʻole i ke ʻano. ʻO ka huina o 4, 72, 168 a me 240 mau hola ma hope o ka lawelawe ʻana i ka lāʻau lapaʻau, ua hoʻoholo ʻia ka waiwai i hoʻohui ʻia o nā metabolites lāʻau no ka nānā ʻana.
IPHASE-HepatoCon™, ma ka ʻaoʻao ʻē aʻe, he ʻōnaehana no ka hoʻoulu ʻana i nā hepatocytes kanaka pili mua i ka hau ma ka HepatoCon™ medium. Ma hope o ka ho'ōla houʻana i ke kanaka muahepatocytes, ua hana 'ia ka ho'ā'o ho'oponopono lā'au ma hope o 3 mau lā o ka mo'omeheu ma ka HepatoCon™ wale nō, a ua like ka manawa o ka ho'oponopono 'ana, ka'ōnaehana mo'omeheu a me ke ala ho'ā'o me nā mea o ka pūnaewele HepatoMax™.
I kēia manawa, no ka hōʻoia ʻana i ka pololei a me ka paʻakikī o nā hopena, ua hana ʻo IPHASE i kahi mana medium (cell-free) a me kahi mana stromal cell i ka manawa like, me ʻekolu replications, a ua hōʻike nā hopena i kaIPHASE-HepatoMax™ aIPHASE-HepatoCon™ Ua hoʻopuka nā ʻōnaehana i kahi pane metabolite kiʻekiʻe ikaika mai ka lā 3 ma mua, a ua hōʻike lāua ʻelua i ka nui o ka huli ʻana o ka metabolic.
- (2) Hoʻopili nui ʻia ka metabolism lāʻau i ka hepatocyteCYP450ʻōnaehana enzyme, a hiki ke ana ʻia kāna hana no ka nānā maikaʻi ʻana.
ʻO kaIPHASE-HepatoMax™ aIPHASE-HepatoCon™ He kūpono kūpono nā ʻōnaehana.
Hoʻonohonoho ʻo IPHASE i ʻehā mau ʻāpana o ka mana waena (ʻaʻohe cell), HepatoMax™, HepatoCon™ a me nā cell stromal. Ke hoʻohana nei i ka pā-pili kanaka muahepatocytes e like me ka laʻana, ka metabolite hoʻonui waiwai oCYP1A2, CYP2B6aCYP3A4ua hōʻoia ʻia i ka lā 0, 3 a me 7 ma hopehepatocytehoʻihoʻi hou no 3 mau lā me ka hoʻohana ʻana i ka finasteride, bupropion a me midazolam ma ke ʻano he substrates. Ua hōʻike ʻia nā waiwai hoʻonui metaboliteIPHASE-HepatoMax™ aIPHASE-HepatoCon™ hōʻike nā ʻōnaehana i ka hana enzyme ikaika i ka lā 7, e hoʻoikaika mau ana i ka metabolism lāʻau maʻamau.
- (3) Ma waho aʻe o ka hōʻoia ʻana o ka hana enzyme, ʻo ke kūpaʻa metabolic maikaʻi kekahi mea koʻikoʻi a koʻikoʻi i ka hōʻoia ʻana i ka pono o nā hiʻohiʻona in vitro.
Ua hoʻonohonoho ʻo IPHASE i ʻekolu mau ʻāpana o HepatoMax™, HepatoCon™ a me nā stromal cell e hoʻoholo ai i ke kūpaʻa metabolic o Tolbutimide ma kahi manawa o 80 mau hola me ka hoʻohana ʻana i nā hepatocytes kanaka mua ma ke ʻano he laʻana, i lawelawe ʻia i 3 mau lā ma hope o ka hoʻōla ʻana o ka mea mua.hepatocytes, a ua hōʻike ʻia nā hopena ʻeluaIPHASE-HepatoMax™ aIPHASE-HepatoCon™ Ua hōʻike nā ʻōnaehana ʻelua i ka hana hoʻomaʻemaʻe metabolic ikaika i loko o ka waiwai lōʻihi o ka manawa assay.
I ka hōʻuluʻulu ʻana, ua hoʻomohala ʻo IPHASE, ma ke ʻano he alakaʻi i ka in vitro bio-reagentsIPHASE HepatoMax™ aIPHASE HepatoCon™ nā kumu hoʻohālike in vitro e pili ana i ka pā-i hoʻohana mua ʻia i ke kanaka, ka monkey, ka ʻīlio, ka ʻiole a me ka ʻiolehepatocytes a me stromal cell o na genera like 'ole, me ka maikai metabolic turnover, active enzyme hana a me ka ikaika metabolic clearance hana, e haawi hou koho no ko makou mea kūʻai mai e hoʻomōhala haʻahaʻa - ʻO kaIPHASE HepatoMax™ aIPHASE HepatoCon™ hāʻawi nā kumu hoʻohālike in vitro i ka nui o ka huli ʻana o ka metabolic, nā hana enzyme ikaika a me nā hana hoʻomaʻemaʻe metabolic ikaika, e hāʻawi ana i nā koho hou no nā mea kūʻai aku e hoʻomohala i nā lāʻau lapaʻau haʻahaʻa, a he hoa nui i ka ʻike lāʻau!
Kuhikuhi:
[1]阮婷婷,鞠武建,熊海伟,等.低清除率药物的代谢稳定性预测模型研究进展[J].中国药科大学学报, 2019(2):152-160.
[2] Di L, Trapa P, Obach R S, et al. He ala hoʻolele hou no ka hoʻoholo ʻana i nā waiwai hoʻomaʻemaʻe haʻahaʻa[J]. ʻO ka metabolism a me ke ʻano o ka lāʻau lapaʻau, 2012, 40(9): 1860-1865.
Ka manawa hoʻouna: 2024-07-19 17:20:41