In Vitro Evaluation of In Vitro Drug Transporter: E ʻimi ana i nā kuleana o ABC Vesicles a me SLC Transporter Cells

Hua'ōlelo: ATP-paʻa cassette(ABC), ABC Transporter, SLC Transporter, Membrane Vesicle, MDR1(P-gp), BSEP, BCRP, MATE1, MATE2-K, OAT1, OATP1B1, MDCK II, Caco-2, Transporter Inhibition, Transporter Substrate Identification ma ka ICHK9id Studies ma ka lāʻau lapaʻau ICHK12 Dra ICHK9ID. MAKE, MAKE SLC Mea lawe.

Nā huahana IPHASE

Kāpae

Ma nā noiʻi lapaʻau, hoʻohana pinepine ka poʻe maʻi i nā lāʻau lapaʻau i ka manawa like, a hiki i kēia mau lāʻau lapaʻau ke hana i ka lāʻau-drug interactions(DDI) i hiki ke hoʻoulu i nā hopena koʻikoʻi a hoʻololi paha i ka maikaʻi o ka mālama ʻana. Hoʻomaka maʻamau ka loiloi DDI me ka hoʻāʻo in vitro e ʻike i nā mea e pili ana i ka waiho ʻana i ka lāʻau e wehewehe i nā mīkini DDI hiki ke loaʻa a loaʻa nā ʻāpana kinetic no ke aʻo hou ʻana. Ua lōʻihi ka manaʻo o ka noiʻi DDI i nā enzyme metabolizing lāʻau. I kēia mau makahiki i hala iho nei, me ka hoʻomohala ʻana o ka ʻenehana biology molecular, ua holomua nui i ka noiʻi ʻana i nā mea lawe lāʻau in vitro, a ʻo ke koʻikoʻi o ka loiloi in vitro o nā DDI e pili ana i ka transporter ua hoʻonui i ka nānā.

ʻO nā mea lawe a me kā lākou mau kuleana

ʻO ka Transporter e pili ana i nā protein transmembrane ma loko o ka membrane cell o nā ʻiʻo like ʻole e hoʻopili ana i ke komo ʻana a me ka puka ʻana o nā mea endogenous a i ʻole nā ​​​​mea exogenous i loko a i waho o nā membrane biological. ʻO ka mea lawe lāʻau lapaʻau in vitro he huaʻōlelo maʻamau no nā protein e lawe i nā lāʻau lapaʻau ma ke ʻano he substrates, aia ma ka ʻili o ka membrane cell o nā ʻiʻo a i ʻole nā ʻāpana, a lawe i ka hana o ka lawe ʻana i ka lāʻau lapaʻau transmembrane, ka mea nui me nā ʻohana ʻelua: ka ATP-cassette paʻa (ABC)superfamily a me solute carrier superfamily (SLC transport), ka mea i hoʻomalu i ke kaʻina lawe lāʻau transmembrane.

ʻO nā mea halihali ABChe ATP-mea halihali e hoʻoneʻe ikaika i nā substrate like ʻole (e laʻa, nā ion, lipids, a me nā lāʻau lapaʻau) ma waena o nā membrane cell, pinepine e kūʻē i nā gradients concentrate. Hoʻokani lākou i nā kuleana koʻikoʻi i ka pale ʻana i ka lāʻau, ka metabolism, a me ka detoxification cellular.Nā mea lawe SLCʻO ka hapa nui o nā mea hoʻokele lua a i ʻole nā ​​mea hoʻokele maʻalahi e hoʻoponopono i ka neʻe ʻana o nā solutes e like me ka glucose, amino acids, a me nā neurotransmitters. ʻAʻole like me nā mea halihali ABC, ʻaʻole lākou e koi pololei iā ATP akā hilinaʻi i nā gradients ion.

ʻO nā mea lawe e hoʻokae i ka lāʻau efflux ka nui o ka P-glycoprotein (P-gp), a i ʻole multi-drug resistance 1 protein (MDR1), breast cancer resistance protein (BCRP). He lālā lākou o ka ʻohana transporter ATP-binding cassette(ABC), e hoʻohana ana i ka ikehu o ka ATP hydrolyzed no ka lawe ʻana i nā lāʻau lapaʻau a me nā mea endogenous. Hiki i nā mea lawe i ke komo ʻana o nā lāʻau lapaʻau i loko o ke keena ke lawe i nā substrates i ka pae i manaʻo ʻia e hoʻokō i nā hopena pharmacological, a no nā lālā o ka ʻohana solute transporter, ʻo ia hoʻi me Organic anion transporting polypeptide (OATPs), Organic anion transporter (OATs), Multidrug and toxin extrusion proteins (MATEs), Organic cation transporter (OCT).

ʻO nā mea lawe nui a me kā lākou mau kuleana:

• -BSEP(Paʻakai Bile Export Pump):He mea koʻikoʻi no ka huna ʻana o ka waikawa bile i loko o ke ake, pili ka BSEP dysfunction i ka maʻi ate cholestatic. ʻO ka lāʻau-induced --BEP hiki ke hoʻoulu i ka hepatotoxicity.
• -BCRP(Polokina Kū'ē i ka maʻi maʻi umauma):Hoʻopilikia ka BCRP i ka bioavailability o nā chemotherapeutics a me nā antivirals, ka lawe ʻana i nā substrates mai nā cell a me ka hoʻoulu ʻana i ka placental a me ke koko-brain barrier penetration.
• -MATE1/MATE2-K(Multidrug and Toxin Extrusion Proteins):Aia i loko o ka renal a me ka ʻiʻo hepatic, MATE1 a me MATE2-K hoʻokuʻu i nā lāʻau cationic, hana synergistically me OCT2 e hoʻoponopono i ka huna ʻana o ka renal.
• -OATP1B1(SLCO1B1):ʻO kahi transporter uptake hepatic koʻikoʻi no ka hoʻomaʻemaʻe statin. He kuleana koʻikoʻi ka OATP1B1 i ka hoʻopili ʻana i nā pūhui endogenous e like me bilirubic acid, bilirubin, steroid-coupled compound, a me nā hormones thyroid, a me ka hoʻomaʻemaʻe ʻana o nā ʻano lāʻau lapaʻau e like me statins, antibiotics, antivirals, a me nā lāʻau anticancer.
• -OAT1(SLC22A6):Hoʻopili i ka lawe ʻana o ka renal i nā anion, me nā antivirals a me nā NSAID, e hoʻopilikia ana i ka pilikia nephrotoxicity.

Hoʻopaʻa ʻia ʻo Transporter a me DDI

Kāohi mea lawehe mea koʻikoʻi o ka hoʻomohala ʻana i ka lāʻau. He kuleana koʻikoʻi ka pāpā ʻana o Transporter i ka lāʻau-drug interactions (DDI) ma o ka hoʻopili ʻana i ka absorption, ka hāʻawi ʻana, ka metabolism, a me ka excretion o nā lāʻau lapaʻau i lawelawe ʻia. Hiki i ka pale ʻana i nā mea lawe efflux e like me MDR1(P-gp) a me BCRP ke hōʻemi i ka extrusion cellular o nā lāʻau lapaʻau, hiki ke alakaʻi i ke kiʻekiʻe intracellular a me nā ʻōnaehana ʻōnaehana. Pēlā nō, hiki i ka pale ʻana i nā mea lawe uptake e like me OATP1B1 ke hōʻemi i ka hoʻomaʻemaʻe ʻana i ka lāʻau hepatic, aʻo ka pale ʻana o BSEP hiki ke hoʻopilikia i ka lawe ʻana i ka bile acid, e hoʻonui ana i ka pilikia o ka cholestasis. Hoʻohui hou, hiki i nā mea lawe e like me MATE1, MATE2-K, a me OAT1, he mea koʻikoʻi no ka hoʻopau ʻana i ka lāʻau lapaʻau renal, ke kāohi ʻia, hiki ke hoʻololi i ka excretion renal a kōkua i ka hōʻiliʻili lāʻau. ʻO nā hiʻohiʻona in vitro, meMDCK IIaKako-2nā laina kelepona, hoʻohana nui ʻia e loiloi i kēia mau pilina transporter, e hāʻawi ana i nā ʻike koʻikoʻi i ka hiki ke DDI i ka wā o ka hoʻomohala ʻana i ka lāʻau.

Nā Hoʻohālike In Vitro no nā Haʻawina Hoʻokele

ʻO ka hoʻomaopopo ʻana i ka hana transporter a me ka wānana ʻana i ka transporter-mediated DDIs hilinaʻi i nā hiʻohiʻona kelepona kūikawā. Hoʻohana nā mea noiʻiHEK293 HOOLAHAnā pūnaewele ma ke ʻano he mana maikaʻi ʻole, e hōʻoia i ka lawe ʻia ʻana ma muli o ka transporter kikoʻī. Hāʻawi ʻo HEK293 MOCK i kahi ʻano like ʻole, ʻoiai ʻo HEK293 MOCK ke kikowaena no ka hōʻoia ʻana i ka kikoʻī hoʻokolohua. Hoʻohana kekahi ʻano hanavesicles membranemai nā pūnaewele e hōʻike nui ana i nā mea lawe, hoʻokaʻawale pinepine ʻia mai HEK293 MOCK cell. Pēlā nō,Mea lawe hoʻomāʻewaʻewa SLCHe mea koʻikoʻi nā hiʻohiʻona no ka hoʻokaʻawale ʻana i ka hana transporter kikoʻī mai ka lawe ʻana ʻole. Ua hōʻoia ʻo MOCK SLC Transporter assays he transporter-dependent ka neʻe ʻana o ka substrate i ʻike ʻia, a ʻo nā hoʻokolohua MOCK SLC Transporter e hoʻoikaika i ka hilinaʻi o ke kumu hoʻohālike. ʻO kēia ala ʻelua e hoʻohana ana i ka HEK293 MOCK a me MOCK SLC Transporter e hoʻonui i ko mākou hiki ke aʻo i ka ATP-ka halihali hilinaʻi ʻole me ka hoʻopilikia ʻole ʻana mai ka metabolism cellular.

ʻO ka laina kelepona ʻo Caco-2, i loaʻa mai ka maʻi maʻi maʻi kanaka, he kumu hoʻohālike ʻē aʻe i hoʻohana ʻia e hoʻohālikelike i ka absorption o ka ʻōpū. ʻO kona hiki ke hoʻokaʻawale i loko o nā pūnaewele enterocyte-like e hōʻike ana i nā mea lawe like ʻole - e like me P - gp a me BCRP - he mea waiwai nui ia no ka loiloi ʻana i ka permeability o ka lāʻau a me nā pilina pili.

Eia kekahi, transporter-specific models like Human BCRP Expressing cell e hiki ai i ka hoʻokolokolo 'ana i ke kiko'ī o ka substrate, ke ʻano kinetic o ka lawe ʻana i ka lāʻau lapaʻau, a me ka nui o ka mea hiki ke hoʻololi i ka hana transporter. Ke hui pū ʻia me ka ʻikepili mai nā hoʻokolohua vesicular, hāʻawi kēia mau ʻōnaehana kelepona i nā ʻike piha i ka dynamics o transporter-mediated drug disposition.

Ka hopena

I ka hōʻuluʻulu ʻana, he mea nui ka hoʻomaopopo ʻana i ka pilina o nā transporter no ka hoʻonui ʻana i ka palekana a me ka pono o ka lāʻau. ʻO nā mea lawe e like me MDR1(P-gp), BCRP, a me BSEP ma ka ʻaoʻao o nā mea lawe uptake e like me OATP 1B1 a me OAT1 slc Transporter e hana koʻikoʻi i nā pharmacokinetics lāʻau. ʻO ka hoʻohana ʻana i nā hiʻohiʻona in vitro me MDCK II, Caco-2, Human BCRP Expressing cells, a me Human MDR1 Knockin MDCK II Cells e hāʻawi i kahi ʻano paʻa no ka hoʻomaopopo ʻana pehea e hiki ai i ka mea hoʻoneʻe transporter ke hoʻopili i nā pilina lāʻau-lāʻau a me ke ʻano o ka lāʻau lapaʻau. He waiwai nui kēia mau ʻike no ka hoʻonui ʻana i ka maikaʻi o ka lāʻau lapaʻau a me ka palekana i nā hoʻonohonoho lapaʻau.