Tau qhov twg los: Medical Micro

Tom qab COVID-19 tshwm sim, ob qhov tshuaj tiv thaiv mRNA tau pom zoo sai sai rau kev lag luam, uas tau nyiam ntau dua rau kev tsim cov tshuaj nucleic acid.Nyob rau hauv xyoo tas los no, ntau cov tshuaj nucleic acid uas muaj peev xwm los ua cov tshuaj blockbuster tau luam tawm cov ntaub ntawv kho mob, npog cov kab mob plawv thiab cov kab mob metabolic, kab mob siab, thiab ntau yam kab mob tsawg.Cov tshuaj Nucleic acid yuav tsum dhau los ua cov tshuaj me me tom ntej thiab cov tshuaj tiv thaiv kab mob.Qhov thib peb hom tshuaj loj tshaj plaws.

Nucleic acid tshuaj qeb

Nucleic acid yog ib qho tshuaj lom neeg macromolecular compound tsim los ntawm polymerization ntawm ntau nucleotides, thiab yog ib qho tseem ceeb tshaj plaws ntawm lub neej.Cov tshuaj Nucleic acid yog ntau yam ntawm oligoribonucleotides (RNA) lossis oligodeoxyribonucleotides (DNA) nrog cov haujlwm sib txawv, uas tuaj yeem ua ncaj qha rau cov kab mob uas ua rau lub hom phiaj cov noob lossis cov hom phiaj mRNAs los kho cov kab mob ntawm qib noob lub luag haujlwm ntawm.

▲ Cov txheej txheem synthesis ntawm DNA rau RNA rau cov protein (Duab qhov chaw: bing)

Tam sim no, cov tshuaj nucleic acid tseem ceeb suav nrog antisense nucleic acid (ASO), me me cuam tshuam RNA (siRNA), microRNA (miRNA), me me activating RNA (saRNA), tub txib RNA (mRNA), aptamer, thiab ribozyme., Antibody nucleic acid conjugated tshuaj (ARC), thiab lwm yam.

Ntxiv rau mRNA, kev tshawb fawb thiab kev loj hlob ntawm lwm cov tshuaj nucleic acid kuj tau txais kev saib xyuas ntau dua nyob rau xyoo tas los no.Hauv 2018, lub ntiaj teb thawj cov tshuaj siRNA (Patisiran) tau pom zoo, thiab nws yog thawj cov tshuaj nucleic acid siv LNP kev xa khoom.Nyob rau hauv xyoo tas los no, kev lag luam ceev ntawm cov tshuaj nucleic acid kuj tau nrawm dua.Hauv 2018-2020 ib leeg, muaj 4 cov tshuaj siRNA, Peb cov tshuaj ASO tau pom zoo (FDA thiab EMA).Tsis tas li ntawd, Aptamer, miRNA thiab lwm qhov chaw kuj tseem muaj ntau yam tshuaj nyob rau hauv kev kho mob theem.

Qhov zoo thiab cov nyom ntawm cov tshuaj nucleic acid

Txij li thaum xyoo 1980, kev tshawb fawb thiab kev tsim cov tshuaj tshiab raws li lub hom phiaj tau maj mam nthuav dav, thiab cov tshuaj tshiab tau pom ntau;cov tshuaj me me-molecule tshuaj thiab cov tshuaj tiv thaiv kab mob ob qho tib si siv cov tshuaj pharmacological los ntawm kev khi rau lub hom phiaj cov protein.Lub hom phiaj proteins tuaj yeem yog Enzymes, receptors, ion channels, thiab lwm yam.

Txawm hais tias cov tshuaj me me muaj qhov zoo ntawm kev tsim khoom yooj yim, kev tswj hwm qhov ncauj, zoo dua cov tshuaj pharmacokinetic, thiab yooj yim dhau los ntawm cov cell membranes, lawv txoj kev loj hlob cuam tshuam los ntawm kev siv tshuaj ntawm lub hom phiaj (thiab seb lub hom phiaj protein puas tsim nyog lub hnab tshos thiab loj)., Qhov tob, polarity, thiab lwm yam);Raws li ib tsab xov xwm hauv Nature2018, tsuas yog 3,000 ntawm ~ 20,000 proteins encoded los ntawm tib neeg genome tuaj yeem yog cov tshuaj, thiab tsuas yog 700 muaj cov tshuaj sib raug tsim (hauv cov tshuaj molecule loj).

Qhov txiaj ntsig loj tshaj plaws ntawm cov tshuaj nucleic acid yog cov tshuaj sib txawv tuaj yeem tsim los ntawm kev hloov lub hauv paus ntawm nucleic acid.Piv nrog rau cov tshuaj uas ua hauj lwm nyob rau hauv cov tsoos protein theem, nws txoj kev loj hlob yog yooj yim, npaum, thiab lom tshwj xeeb;piv nrog genomic DNA-theem kev kho mob, cov tshuaj nucleic acid tsis muaj kev pheej hmoo ntawm kev sib xyaw ntawm cov noob thiab hloov tau yooj yim dua thaum lub sijhawm kho.Cov tshuaj tuaj yeem nres thaum tsis muaj kev kho mob.

Cov tshuaj Nucleic acid muaj cov txiaj ntsig zoo xws li kev ua haujlwm siab, ua haujlwm siab thiab ua haujlwm ntev.Txawm li cas los xij, nrog ntau qhov zoo thiab kev txhim kho nrawm, cov tshuaj nucleic acid tseem tab tom ntsib ntau yam teeb meem.

Ib qho yog RNA hloov kho los txhim kho kev ruaj ntseg ntawm cov tshuaj nucleic acid thiab txo cov immunogenicity.

Qhov thib ob yog kev txhim kho cov neeg nqa khoom kom muaj kev ruaj ntseg ntawm RNA thaum lub sij hawm hloov cov txheej txheem nucleic acid thiab cov tshuaj nucleic acid kom ncav cuag lub hom phiaj ntawm lub hlwb / lub hom phiaj;

Qhov thib peb yog kev txhim kho ntawm kev xa tshuaj.Yuav ua li cas txhawm rau txhim kho cov tshuaj xa mus kom ua tiav cov txiaj ntsig zoo ib yam nrog cov tshuaj tsawg.

Kev hloov kho tshuaj ntawm cov tshuaj nucleic acid

Exogenous nucleic acid tshuaj yuav tsum tau kov yeej ntau yam teeb meem thiaj li nkag mus rau hauv lub cev los ua lub luag haujlwm.Cov teeb meem no kuj ua rau muaj teeb meem hauv kev tsim cov tshuaj nucleic acid.Txawm li cas los xij, nrog rau kev tsim cov thev naus laus zis tshiab, qee qhov teeb meem twb tau daws los ntawm kev hloov kho tshuaj.Thiab kev ua tiav hauv kev xa khoom siv thev naus laus zis tau ua lub luag haujlwm tseem ceeb hauv kev tsim cov tshuaj nucleic acid.

Kev hloov kho tshuaj tuaj yeem txhim kho lub peev xwm ntawm cov tshuaj RNA los tiv thaiv kev degradation los ntawm endogenous endonucleases thiab exonucleases, thiab txhim kho kev ua tau zoo ntawm cov tshuaj.Rau cov tshuaj siRNA, kev hloov kho tshuaj lom neeg kuj tuaj yeem txhim kho qhov kev xaiv ntawm lawv cov tshuaj tiv thaiv kab mob kom txo qis kev ua haujlwm ntawm RNAi, thiab hloov lub cev thiab tshuaj lom neeg los txhim kho kev xa khoom.

1. Kev hloov kho cov suab thaj

Nyob rau theem pib ntawm kev txhim kho cov tshuaj nucleic acid, ntau cov nucleic acid tebchaw tau nthuav tawm cov haujlwm zoo hauv vitro, tab sis lawv cov haujlwm hauv vivo tau txo qis lossis ploj tag nrho.Qhov laj thawj tseem ceeb yog qhov tsis hloov pauv nucleic acids tau yooj yim tawg los ntawm enzymes lossis lwm yam tshuaj endogenous hauv lub cev.Kev hloov kho cov suab thaj feem ntau suav nrog kev hloov pauv ntawm 2-txoj hauj lwm hydroxyl (2'OH) ntawm qab zib rau methoxy (2'OME), fluorine (F) lossis (2'MOE).Cov kev hloov kho no tuaj yeem ua tiav qhov kev ua si thiab kev xaiv, txo cov teebmeem ntawm lub hom phiaj, thiab txo cov kev mob tshwm sim.

▲ Kev hloov kho cov suab thaj (daim duab qhov chaw: siv 4)

2. Phosphoric acid pob txha hloov kho

Feem ntau siv tshuaj hloov kho ntawm cov pob txha phosphate yog phosphorothioate, uas yog, tsis muaj cov pa oxygen hauv cov pob txha phosphate ntawm cov nucleotide yog hloov nrog sulfur (PS hloov kho).PS kev hloov kho tuaj yeem tiv thaiv qhov degradation ntawm nucleases thiab txhim kho kev sib cuam tshuam ntawm cov tshuaj nucleic acid thiab plasma proteins.Binding peev, txo lub raum clearance tus nqi thiab nce ib nrab-lub neej.

▲ Kev hloov pauv ntawm phosphorothioate (daim duab qhov chaw: siv 4)

Txawm hais tias PS tuaj yeem txo qhov kev sib raug zoo ntawm nucleic acids thiab lub hom phiaj cov noob, PS kev hloov kho ntau dua hydrophobic thiab ruaj khov, yog li nws tseem yog ib qho kev hloov pauv tseem ceeb hauv kev cuam tshuam nrog me me nucleic acids thiab antisense nucleic acids.

3. Kev hloov kho ntawm tsib lub nplhaib ntawm ribose

Kev hloov kho ntawm tsib lub nplhaib ntawm ribose yog hu ua kev hloov kho tshuaj thib peb, suav nrog kev sib txuas ntawm nucleic acid-locked nucleic acid BNAs, peptide nucleic acid PNA, phosphorodiamide morpholino oligonucleotide PMO, cov kev hloov kho no tuaj yeem txhim kho cov tshuaj nucleic acid ntxiv, kev txhim kho tshwj xeeb, thiab lwm yam.

4. Lwm yam kev hloov kho tshuaj

Hauv kev teb rau cov kev xav tau sib txawv ntawm cov tshuaj nucleic acid, cov kws tshawb fawb feem ntau ua rau kev hloov kho thiab kev hloov pauv ntawm cov hauv paus thiab nucleotide chains kom nce kev ruaj ntseg ntawm cov tshuaj nucleic acid.

Txog tam sim no, tag nrho cov tshuaj RNA-targeting cov tshuaj tau pom zoo los ntawm FDA yog chemically engineered RNA analogs, txhawb kev siv hluav taws xob hloov kho tshuaj.Ib leeg-stranded oligonucleotides rau cov tshuaj hloov kho tshwj xeeb sib txawv tsuas yog nyob rau hauv ib theem, tab sis lawv txhua tus muaj cov khoom zoo sib xws ntawm lub cev thiab tshuaj lom neeg, thiab yog li ntawd muaj cov tshuaj pharmacokinetics thiab cov khoom siv roj ntsha.

Kev xa khoom thiab kev tswj hwm cov tshuaj nucleic acid

Cov tshuaj nucleic acid uas tsuas yog siv rau kev hloov kho tshuaj tsuas yog tseem yooj yim degraded sai sai hauv cov ntshav ncig, tsis yooj yim rau hauv cov ntaub so ntswg, thiab tsis yooj yim rau kev nkag mus rau lub hom phiaj cell membrane kom ncav cuag qhov chaw ntawm kev ua hauv cytoplasm.Yog li ntawd, lub zog ntawm kev xa khoom yog xav tau.

Tam sim no, nucleic acid cov tshuaj vectors feem ntau muab faib ua kab mob thiab tsis yog kab mob.Cov qub muaj xws li adenovirus-associated virus (AAV), lentivirus, adenovirus thiab retrovirus, thiab lwm yam. Cov no suav nrog lipid carriers, vesicles thiab lwm yam.Los ntawm kev xav ntawm cov tshuaj lag luam, cov kab mob kis kab mob thiab cov neeg nqa khoom lipid yog cov paub tab ntau dua hauv kev xa cov tshuaj mRNA, thaum cov tshuaj me me nucleic acid siv ntau cov neeg nqa khoom lossis cov cuab yeej thev naus laus zis xws li liposomes lossis GalNAc.

Txog rau tam sim no, feem ntau cov kev kho mob nucleotide, suav nrog yuav luag tag nrho cov tshuaj nucleic acid tau pom zoo, tau siv hauv zos, xws li lub qhov muag, qaum qaum, thiab daim siab.Nucleotides feem ntau yog hydrophilic polyanions loj, thiab cov cuab yeej no txhais tau hais tias lawv tsis tuaj yeem yooj yim dhau los ntawm cov plasma membrane.Nyob rau tib lub sijhawm, oligonucleotide-raws li cov tshuaj kho mob feem ntau tsis tuaj yeem hla cov hlab ntsha hauv hlwb (BBB), yog li kev xa mus rau lub hauv paus paj hlwb (CNS) yog qhov kev sib tw tom ntej rau cov tshuaj nucleic acid.

Nws yog ib qho tsim nyog sau cia tias nucleic acid raws li kev tsim thiab kev hloov kho nucleic acid tam sim no yog qhov tseem ceeb ntawm kev saib xyuas ntawm cov kws tshawb fawb hauv cheeb tsam.Rau kev hloov kho tshuaj, tshuaj hloov kho nucleic acid, uas tsis yog-natural nucleic acid sequence tsim los yog txhim kho, nucleic acid muaj pes tsawg leeg, vector siv, nucleic acid synthesis txoj kev, thiab lwm yam. Cov kev kawm yog feem ntau patentable daim ntawv thov kev kawm.

Coj tus kabmob tshiab no ua piv txwv.Txij li thaum nws RNA yog ib yam khoom uas muaj nyob rau hauv lub ntuj tsim, "RNA ntawm tus kab mob tshiab" nws tus kheej tsis tuaj yeem tso cai patent.Txawm li cas los xij, yog tias tus kws tshawb fawb tshawb fawb cais lossis rho tawm RNA lossis cov khoom tawg uas tsis paub hauv tshuab los ntawm tus kabmob tshiab thawj zaug thiab siv nws (piv txwv li, hloov nws mus rau hauv cov tshuaj tiv thaiv), ces ob qho tib si nucleic acid thiab tshuaj tiv thaiv tuaj yeem tso cai patent txoj cai raws li txoj cai.Tsis tas li ntawd, cov khoom tsim tawm los ntawm nucleic acid molecules hauv kev tshawb fawb ntawm tus kabmob tshiab no, xws li primers, probes, sgRNA, vectors, thiab lwm yam, yog txhua yam khoom siv patentable.

Cov lus xaus

Sib txawv ntawm cov txheej txheem ntawm cov tshuaj me me molecule tshuaj thiab cov tshuaj tiv thaiv kab mob, cov tshuaj nucleic acid tuaj yeem txuas ntxiv kev tshawb nrhiav tshuaj mus rau qib caj ces ua ntej cov proteins.Nws yog qhov pom tau tias nrog kev nthuav dav ntxiv ntawm cov lus qhia thiab kev txhim kho txuas ntxiv ntawm kev xa khoom thiab kev hloov kho thev naus laus zis, cov tshuaj nucleic acid yuav nrov rau cov neeg mob ntau dua thiab tiag tiag los ua lwm chav kawm ntawm cov khoom tawg tom qab cov tshuaj molecule me me thiab cov tshuaj tiv thaiv kab mob.

Cov ntaub ntawv siv:

1.http://xueshu.baidu.com/usercenter/paper/show?paperid=e28268d4b63ddb3b22270ea1763b2892&site=xueshu_se

2.https://www.biospace.com/article/releases/wave-life-sciences-announces-initiation-of-dosing-in-phase-1b-2a-focus-c9-clinical-trial-of-wve-004-in-amyotrophic-lateral-sclerosis-and-frontiampo.

3. Liu Xi, Sun Fang, Tao Qichang;Txawj Ntse."Kev tshuaj xyuas ntawm patentability ntawm cov tshuaj nucleic acid"

4. CICC: cov tshuaj nucleic acid, lub sijhawm tau los

Yam khoom:

Cell Direct RT-qPCR cov khoom siv

Mouse Tail Direct PCR cov khoom siv

Tsiaj Tissue Direct PCR cov khoom siv