In Vitro Evaluation ea In Vitro Drug Transporter: Ho Lekola Mesebetsi ea ABC Vesicles le SLC Transporter Cells

Mantsoe a bohlokoa: ATP-cassette e tlamang(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 ICHda Ident Studies Draft1 HEK293 MOCK, MOTŠOA SLC Sepalangoang.

IPHASE Lihlahisoa

Ka morao

Lits'ebetsong tsa tleliniki hangata bakuli ba sebelisa litlhare tse ngata ka nako e le 'ngoe,' me meriana ena e ka hlahisa tšebelisano ea lithethefatsi-lithethefatsi (DDI) tse nang le monyetla oa ho baka litlamorao tse mpe kapa tsa fetola katleho ea kalafo. Tekolo ea DDI hangata e qala ka tlhahlobo ea in vitro ho fumana lintlha tse ka amang ts'ebeliso ea lithethefatsi ho hlakisa mekhoa e ka bang teng ea DDI le ho fumana litekanyetso tsa kinetic bakeng sa boithuto bo bong. Li-enzyme tsa metabolizing ea lithethefatsi haesale e le sepheo sa lipatlisiso tsa DDI. Lilemong tsa morao tjena, ka nts'etsopele ea theknoloji ea baeloji ea limolek'hule, ho entsoe tsoelo-pele e kholo thutong ea lipalangoang tsa lithethefatsi tsa in vitro, 'me bohlokoa ba tlhahlobo ea in vitro ea li-DDI tse thehiloeng ho li-transporter e hohetse tlhokomelo e ntseng e eketseha.

Bapalami le Mesebetsi ea Bona

Transporter e bua ka liprotheine tsa transmembrane ho pholletsa le lera la lisele tsa lisele tse fapaneng tse hokahanyang ho kena le ho tsoa ha lintho tsa tlhaho kapa tse tsoang kantle ho lera tsa tlhaho. In vitro drug transporter ke lentsoe le akaretsang bakeng sa liprotheine tse nkang lithethefatsi joalo ka li-substrates, tse teng holim'a lisele tsa lisele kapa litho tsa 'mele,' me li nka mosebetsi oa ho tsamaisa lithethefatsi tsa transmembrane, haholo-holo malapa a mabeli: the ATP-khasete e tlamang(ABC)superfamily and solute carrier superfamily (SLC transport), e laolang ts'ebetso ea ho tsamaisa lithethefatsi ea transmembrane.

Bapalami ba ABCke lipalangoang tse itšetlehileng ka ATP tse tsamaisang li-substrates tse fapaneng (mohlala, li-ion, lipids, le lithethefatsi) ho pholletsa le lisele tsa lisele, hangata khahlanong le likhahla tsa mahloriso. Ba phetha karolo ea bohlokoa ho hanyetsang lithethefatsi, metabolism, le detoxification ea lisele.Bapalami ba SLChangata ke lipalangoang tsa bobeli tse sebetsang kapa tse nolofalitsoeng tse hokahanyang motsamao oa metsoako e kang tsoekere, li-amino acid le li-neurotransmitters. Ho fapana le lipalangoang tsa ABC, ha li hloke ATP ka kotloloho empa li itšetleha ka li-gradients tsa ion.

Lipalangoang tse kenellang ho efflux ea lithethefatsi li kenyelletsa P-glycoprotein (P-gp), kapa protheine e 1 ea ho hanyetsa lithethefatsi (MDR1), protheine ea ho hanyetsa mofetše oa matsoele (BCRP). Ke litho tsa lelapa la bapalami ba ATP-e tlamang lik'hasete(ABC), ba sebelisang matla a ATP ea hydrolyzed bakeng sa ho tsamaisa lithethefatsi le lintho tsa tlhaho. Lipalangoang tse hokahanyang ho kena ha lithethefatsi ka seleng li ka nka li-substrates ho ea sebakeng se lebisitsoeng ho fana ka liphello tsa meriana, 'me ke litho tsa lelapa la solute transporter, haholo-holo ho akarelletsa le Organic anion transporting polypeptide (OATPs), Organic anion transporter (OATs), Multidrug le chefo extrusion proteins (MATEs), Organic transporter (OCTs) joalo-joalo.

Lipalangoang tsa Bohlokoa le Mesebetsi ea tsona:

• -BSEP(Pompi e Romellang Letsoai la Bile):E bohlokoa bakeng sa secretion ea bile acid sebeteng, ho se sebetse ha BSEP ho amahanngoa le lefu la sebete la cholestatic. Lithethefatsi-induced -- Thibelo ea BSEP e ka baka hepatotoxicity.
• -BCRP(Protheine e hanyetsanang le Kankere ea Matsoele):BCRP e ama bioavailability ea chemotherapeutics le antivirals, ho romela li-substrates ho tsoa liseleng le ho susumetsa placental le mali-ho kenella ha boko.
• -MATE1/MATE2-K(Liprotheine tsa Multidrug le Toxin Extrusion):E fumaneha ka har'a lisele tsa renal le hepatic, MATE1 le MATE2-K e ntša lithethefatsi tsa cationic, e sebetsa ka mokhoa o ts'oanang le OCT2 ho lokisa ho tsoa ha renal.
• -OATP1B1(SLCO1B1):Sepalangoang sa hepatic uptake se bohlokoa bakeng sa ho tlosoa ha statin. OATP1B1 e bapala karolo ea bohlokoa ho kopanyelletsa ho nkeloa ha metsoako ea endo native joalo ka bilirubic acid, bilirubin, steroid-completed compounds, le lihomone tsa qoqotho, hammoho le ho hlakisa sebete sa mefuta e mengata ea litlhare tse kang li-statins, lithibela-mafu, li-antiviral le li-anticancer.
• -OAT1(SLC22A6):E kenella ho kenngoa ha renal ea anions, ho kenyelletsa le li-antiviral le li-NSAID, tse susumetsang kotsi ea nephrotoxicity.

Thibelo ea Lipalangoang le DDI

Thibelo ea lipalangoangke karolo ea bohlokoa ea ntlafatso ea lithethefatsi. Thibelo ea lipalangoang e bapala karolo ea bohlokoa litšebelisanong tsa lithethefatsi-lithethefatsi (DDI) ka ho ama ho monya, kabo, metabolism, le ho ntšoa ha lithethefatsi tse tsamaisoang hammoho. Thibelo ea lipalangoang tsa efflux joalo ka MDR1(P-gp) le BCRP e ka fokotsa ho tsoa ha lithethefatsi ka liseleng, e leng se ka lebisang maemong a phahameng a intracellular le systemic. Ka mokhoa o ts'oanang, ho thibela lipalangoang tse kang OATP1B1 ho ka fokotsa tumello ea lithethefatsi tsa sebete, athe thibelo ea BSEP e ka kena-kenana le lipalangoang tsa bile acid, ea phahamisa kotsi ea cholestasis. Ho feta moo, lipalangoang tse joalo ka MATE1, MATE2-K, le OAT1, tse bohlokoa bakeng sa ho felisa lithethefatsi tsa renal, ha li thibetsoe, li ka fetola ho ntšoa ha refi le ho kenya letsoho ho bokellaneng ha lithethefatsi. Mefuta ea in vitro, ho kenyelletsaMDCK IIleKako-2lisele, li sebelisoa haholo ho lekola litšebelisano tsena tsa lipalangoang, ho fana ka lintlha tsa bohlokoa mabapi le bokhoni ba DDI nakong ea nts'etsopele ea lithethefatsi.

In Vitro Models bakeng sa Lithuto tsa Transporter

Ho utloisisa tšebetso ea sepalangoang le ho bolela esale pele sepalangoang-li-DDI tse mahareng li itšetleha ka mefuta e ikhethileng ea lisele. Bafuputsi baa sebelisaHEK293 MOSOTHOlisele e le taolo e mpe, ho etsa bonnete ba hore sepalangoang leha e le sefe se hlokometsoeng se bakoa ke sepalangoang se itseng. HEK293 MOCK e fana ka semelo se ts'oanang, ha HEK293 MOCK e ntse e le bohareng ba ho netefatsa lintlha tsa liteko. Mokhoa o mong oa sebetsali-membrane vesiclesho tloha liseleng tse tsamaisang thepa ka mokhoa o feteletseng, hangata li arohane le lisele tsa HEK293 MOCK. Ka mokhoa o ts'oanang,soma SLC Sepalangoangmefuta e bohlokoa bakeng sa ho khetholla ts'ebetso e khethehileng ea lipalangoang ho tsoa ho tse sa khethoeng. Liteko tsa MOCK SLC Transporter li tiisa hore motsamao oa substrate o hlokometsoeng o itšetlehile ka sepalangoang, 'me liteko tsa MOCK SLC Transporter li tiisa ho tšepahala ha mohlala. Mokhoa ona oa bobeli o sebelisang HEK293 MOCK le MOCK SLC Transporter o ntlafatsa bokhoni ba rona ba ho ithuta ATP-sepalangoang se itšetlehileng ka sona ntle le tšitiso ea metabolism ea cellular.

Caco-2 cell line, e nkiloeng ho motho colon carcinoma, ke mohlala o mong o tloaelehileng o sebelisoang ho etsisa ho monya lithethefatsi ka mala. Bokhoni ba eona ba ho ikarola ka li-enterocyte-joaloka lisele tse hlahisang lipalangoang tse fapaneng-joalo ka P-gp le BCRP-e etsa hore e be ea bohlokoa haholo bakeng sa ho lekola bokhoni ba lithethefatsi le likamano tse ka bang teng.

Ho feta moo, li-transporter- mefuta e ikhethileng joalo ka Lisele tsa Human BCRP Expressing li thusa ho etsa lipatlisiso tsa sebopeho sa substrate, boits'oaro ba kinetic ba ho tsamaisa lithethefatsi, le hore na li-inhibitors li ka fetola tšebetso ea sepalangoang hakae. Ha li kopantsoe le lintlha tse tsoang litlhahlobong tsa vesicular, litsamaiso tsena tsa cellular li fana ka lintlha tse felletseng mabapi le matla a transporter-mediated drug disposition.

Qetello

Ka bokhuts'oane, ho utloisisa litšebelisano tsa li-transporter ho bohlokoa bakeng sa ho ntlafatsa polokeho le katleho ea lithethefatsi. Lipalangoang tse kang MDR1(P-gp), BCRP, le BSEP hammoho le lipalangoang tse tsamaisang thepa joalo ka OATP 1B1 le OAT1 slc Transporter li bapala karolo ea bohlokoa ho pharmacokinetics ea lithethefatsi. Tšebeliso ea mefuta ea in vitro e kenyeletsang MDCK II, Caco-2, Human BCRP Expressing cells, le Human MDR1 Knockin MDCK II Cells e fana ka moralo o matla oa ho utloisisa hore na thibelo ea lipalangoang e ka ama litšebelisano tsa lithethefatsi le lithethefatsi le mokhoa oa ho sebelisa lithethefatsi ka kakaretso. Lintlha tsena li bohlokoa haholo bakeng sa ho ntlafatsa ts'ebetso ea lithethefatsi le polokeho maemong a bongaka.