Ntawm Prisys Biotechnology, peb lees paub qhov kev txaus siab loj hlob ntawm oligonucleotide-raws li tshuaj, tshwj xeeb tshaj yog kev cuam tshuam me me RNA (siRNA) kev kho mob. siRNA sawv cev rau tiam thib peb ntawm cov tshuaj kho mob tom qab cov tshuaj molecule me thiab cov tshuaj tiv thaiv kab mob -raws li kev kho mob, thiab tau pom muaj peev xwm loj hauv kev hloov pauv ntawm kev txhim kho tshuaj.
siRNA Mechanism of Action:
siRNAs yog luv nucleotide sequences ntawm txog 20 lub hauv paus uas siv Watson-Crick base pairing cov hauv paus ntsiab lus los khi rau ntxiv mRNA strands, yog li cuam tshuam rau cov txheej txheem sau thiab txhais lus. Raws li lawv cov qauv thiab kev hloov pauv, siRNAs raug cais raws li antisense oligonucleotides (ASO), me me cuam tshuam RNAs (siRNA), me me ua kom RNAs (saRNA), microRNAs (miRNA), thiab aptamers. Ntawm cov no, siRNA tau txais kev saib xyuas tseem ceeb vim nws qhov kev cog lus kho tau zoo thiab kev ua haujlwm thev naus laus zis.
Qhov tseem ceeb ntawm kev hloov kho tshuaj thiab kev xa khoom:
Unmodified siRNAs muaj ib nrab luv luv -lub neej thiab ua rau muaj kev pheej hmoo ntawm - lub hom phiaj toxicity. Kev hloov kho tshuaj thiab cov tshuab xa khoom tsim nyog ua lub luag haujlwm tseem ceeb hauv kev ua tiav cov txiaj ntsig kho mob. Rau cov tshuaj siRNA uas tau pom zoo tau hais ua ntej dhau los tau raug hloov kho tshuaj, xws li phosphorothioate nraub qaum hloov kho thiab hloov kho ribose xws li 2'-O-Methyl (2-Ome), 2'-O-Methoxyethyl (2'OF-{1}). Tsis tas li ntawd, kev ua tau zoo ntawm siRNA kev xa khoom vam khom rau kev xa khoom siab tshaj plaws xws li LNP thiab GalNAc conjugates, ob qho tib si uas tau pom muaj peev xwm ua rau lub siab lub hom phiaj.
Kev Ntsuam Xyuas Kev Nyab Xeeb Preclinical thiab Kev Xaiv Tsiaj Tsiaj:
Kev soj ntsuam kev nyab xeeb hauv tsev kho mob ntawm cov tshuaj siRNA suav nrog kev ntsuas lub hom phiaj - tshwj xeeb thiab tawm - lub hom phiaj toxicity hauv cov qauv tsiaj. Kev xaiv tsiaj txhu, suav nrog cov nas thiab tsis yog - tib neeg primates (NHP), nyob ntawm cov yam ntxwv tshwj xeeb ntawm cov tshuaj siRNA. Piv txwv li, hom primate muaj kev sib raug zoo nrog tib neeg nyob rau hauv cov nqe lus ntawm lub hom phiaj mRNA ib ntus thiab muab kev ntsuam xyuas zoo dua ntawm siRNA pharmacokinetics thiab pharmacodynamics. Kev siv tsis yog -tib neeg primates rau kev ntsuam xyuas kev nyab xeeb yog ib qho kev coj ua, tshwj xeeb tshaj yog rau cov tshuaj siRNA tsom rau lub siab.
Lub siab Toxicity thiab Immune Responses:
Lub siab yog lub cev tseem ceeb ntawm kev txhawj xeeb hauv siRNA kho. Cov tshuaj siRNA tau pom zoo tau pom ntau yam yam ntxwv cuam tshuam nrog rau daim siab - kev xa khoom tshwj xeeb, hepatocyte uptake, thiab muaj peev xwm hepatotoxicity. Cov kev tshawb fawb histopathological tau qhia txog hepatocellular vacuolization, inflammatory infiltration, thiab nce me ntsis hauv daim siab enzymes xws li alanine transaminase (ALT) thiab aspartate transaminase (AST). Cov lus teb tiv thaiv kab mob, suav nrog tshuaj tiv thaiv - tshuaj tiv thaiv kab mob (ADA), cytokine tso tawm, ntxiv kev ua kom muaj zog, thiab kev tiv thaiv kab mob phenotyping, kuj tau raug tshuaj xyuas thiab tshuaj xyuas.
siRNA-kev sib kho gene silencing muaj peev xwm kho tau zoo heev. Txawm li cas los xij, kev ua tiav qhov kev ntsiag to yuav yog qhov nyuaj. Tawm -cov teebmeem tshwm sim, qhov twg siRNA inadvertently suppresses unintended genes, tuaj yeem ua rau muaj qhov tshwm sim tsis tau pom dua. Lub Nyij Pooj Tshuaj thiab Kev Kho Mob Device Administration (PMDA) lees paub qhov no hauv nws "Cov Lus Qhia rau Tsis yog-Kev Ntsuam Xyuas Kev Nyab Xeeb Kev Nyab Xeeb ntawm Oligonucleotide Therapeutic Products," categorizing siRNA toxicity rau hauv hom phiaj thiab tawm - hom phiaj.
Lub siab: Lub Hom Phiaj Prime, Kev txhawj xeeb Prime
Lub siab yog lub hauv paus tseem ceeb ntawm kev txhawj xeeb rau cov tshuaj siRNA. Txhawm rau kom nkag siab tob dua, peb txheeb xyuas rau 6 lub khw muag tshuaj siRNA, tsom mus rau cov yam ntxwv ntawm cov tshuaj, pharmacokinetics, thiab soj ntsuam kab mob. Qhov tseem ceeb, tag nrho rau lub siab lub siab - ntsig txog cov noob rau kev sib txuas lossis kev qhia. Tsis tas li ntawd, lawv tau hloov kho tshuaj, xws li thiophosphate, uas tuaj yeem ua rau hepatotoxicity los ntawm kev sib txuas ntawm cov protein ntau. Ob leeg lipid nanoparticles (LNPs) thiab N-acetylgalactosamine (GalNAc) conjugation pab txhawb lub siab lub hom phiaj. Piv txwv li, 80-90% ntawm LNPs nyob rau hauv Patisiran, ib qho LNP-siRNA tshuaj, tsub kom nyob rau hauv daim siab raws li intravenous tshuaj. GalNAc-siRNAs siv ASGPR-mediated endocytosis rau intracellular uptake, nrog ASGPR qhia tau zoo heev hauv hepatocytes.
Daim siab Toxicity Profile: Los ntawm Kev Ncua Rau Kev Mob
Cov ntaub so ntswg faib tshuaj ntsuam xyuas tas li qhia qhov siab tshaj siRNA concentrations hauv daim siab. Lumasiran, piv txwv li, tau nthuav tawm AUC ntawm 76700 h*ug/mL thiab ib nrab-lub neej ntawm 409h hauv cov liab ntawm 10 mg / kg. Tshwj xeeb tshaj yog, ntev -lub sij hawm liab toxicity kev tshawb fawb qhia koob tshuaj- nyob ntawm qhov hloov pauv ntawm lub siab histopathological:
1. Hepatocyte/Kupffer cell basophilic granules: Cov no yuav sawv cev rau cov tshuaj, nws cov metabolites, los yog cellular adaptation rau oligonucleotides.
2. Hepatocyte vacuolation: Muaj peev xwm ua rau cov nyhuv thib ob vim yog cytoplasmic basophilic granule tsub zuj zuj.
3. Inflammatory infiltration: Cov tshuaj muaj zog ntau tuaj yeem ua rau cov pro-cov kab mob inflammatory.
4. Hepatocyte ib leeg -cell necrosis: Kev puas tsuaj loj tshwm sim ntawm cov koob tshuaj ntau nrog cov basophilic granule ntau.
Kev nce ntshav biochemistry cov cim zoo li ALT thiab AST feem ntau nrog cov kev hloov pauv no. Txawm li cas los xij, kev ntsuam xyuas cov ntaub so ntswg puas tsuaj, cuam tshuam qhov ntsuas kev hloov pauv, thim rov qab, thiab kev tshawb pom zoo sib xws hauv lwm cov tshuaj siRNA yog qhov tseem ceeb rau kev txiav txim siab qhov teeb meem tiag tiag ntawm cov kev hloov no.
Tshaj li lub siab: Tawm-Txoj kev lom thiab tiv thaiv kab mob
Kev sib xyaw ua ke sib xyaw ua ke - ywj siab tawm - lub hom phiaj toxicity kuj tseem ceeb. Kev tiv thaiv kab mob tiv thaiv kab mob, feem ntau nyob ntawm nucleotide kab ke tsim thiab kev hloov kho tshuaj, yog ib qho txiaj ntsig. siRNA tshuaj tuaj yeem ua rau lub cev tsis muaj zog, mus rau qhov chaw txhaj tshuaj (Patiiran, Inclisiran) thiab platelet (PLT) txo qis (Patisiran, Givosiran). Kev ntsuas lub cev tiv thaiv kab mob koom nrog hauv vivo tsom xam ntawm cov cim tseem ceeb thiab kev ntsuas hauv vitro siv cov kab mob peripheral cov ntshav mononuclear cais lossis tag nrho cov ntshav los kwv yees tso tawm cytokine.
Tsis yog -hybridization- cuam tshuam tawm - cov txiaj ntsig ntawm lub hom phiaj, xws li kev hloov pauv ntawm coagulation, kuj tuaj yeem tshwm sim. Qee cov tshuaj siRNA xaiv inhibit Factor IXa thiab cofactor VIIIa hauv coagulation cascade, hloov thrombin lub sijhawm ua haujlwm thiab cuam tshuam tsis ncaj qha rau coagulation (xws li Patisiran, Lumasiran, me ntsis nce APTT). Tsis tas li ntawd, cov tshuaj siRNA tuaj yeem khi rau inhibitor H, ua kom txoj hauv kev ntxiv los ntawm txoj kev. Overactivation ntawm complement tuaj yeem cuam tshuam nws txoj haujlwm, mus rau theem nrab o thiab vasculitis (piv txwv li, Patisiran-induced C3a nce, daim siab inflammatory infiltration, perivascular o ntawm qhov chaw infusion).
Delivery System Toxicity: Tsis txhob saib tus cab kuj
Kev xa khoom, feem ntau LNPs, tsim nyog tau txais kev saib xyuas zoo ib yam. Novel excipients los yog hloov cov khoom sib piv hauv LNPs yuav tsum tau tshawb xyuas kom meej. Thaum cov lus qhia tshwj xeeb LNP tseem tsis tuaj, cov lus qhia uas twb muaj lawm xws li "Kev Taw Qhia rau Kev Lag Luam: Lipiodol Khoom Chemistry, Manufacturing, thiab Tswjhwm; Tib Neeg Pharmacokinetics thiab Bioavailability; Thiab Cov Ntaub Ntawv Sau Npe" tuaj yeem muab cov kev pom zoo. Patisiran's LNP kev tshawb nrhiav kuj tuaj yeem ua tus siv. Qhov tseem ceeb, yog tias LNPs poob rau hauv qhov tsis yog- tus qauv sau npe ywj pheej, lawv tuaj yeem tshawb xyuas nrog siRNA nrog tib hom tsiaj xws li siRNA tsis yog - kev soj ntsuam kev nyab xeeb, xav txog ob - hom kev xav tau rau siRNA.
Xaus:
Lub teb ntawm siRNA kev kho mob txuas ntxiv mus ntxiv nrog rau kev tsom mus rau kev txhim kho kev xa khoom, kev hloov kho tshuaj, txo qis - cov txiaj ntsig ntawm lub hom phiaj, thiab nthuav dav cov lus qhia. Prisys Biotechnology tseem cog lus rau kev tsim kho tshiab siRNA-raws li kev kho mob, siv peb cov kev txawj ntse hauv kev tshawb nrhiav tshuaj thiab kev xa khoom. Peb sib zog ua kom muaj kev hloov pauv hloov kho uas daws cov kev xav tau kev kho mob tsis zoo thiab txhim kho cov txiaj ntsig ntawm tus neeg mob.
