Umthombo: Medical Micro

Ngemuva kokugqashuka kwe-COVID-19, imigomo emibili ye-mRNA yagunyazwa ngokushesha ukuthi ikhangiswe, okudonse ukunakwa okwengeziwe ekuthuthukisweni kwezidakamizwa ze-nucleic acid.Eminyakeni yamuva, izidakamizwa eziningi ze-nucleic acid ezinamandla okuba izidakamizwa ze-blockbuster zishicilele idatha yomtholampilo, ehlanganisa izifo zenhliziyo kanye ne-metabolic, izifo zesibindi, nezinhlobonhlobo zezifo ezingavamile.Izidakamizwa ze-Nucleic acid kulindeleke ukuthi zibe izidakamizwa ze-molecule encane elandelayo kanye nezidakamizwa ze-antibody.Uhlobo lwesithathu olukhulu lwezidakamizwa.

Isigaba somuthi we-Nucleic acid

I-Nucleic acid iyinhlanganisela ye-biological macromolecular eyakhiwe yi-polymerization yama-nucleotide amaningi, futhi ingenye yezinto eziyisisekelo kakhulu zokuphila.Izidakamizwa ze-Nucleic acid ziyi-oligoribonucleotides (RNA) ehlukahlukene (i-RNA) noma i-oligodeoxyribonucleotides (i-DNA) enemisebenzi ehlukene, engenza ngokuqondile izakhi zofuzo ezibangelwa izifo noma iqondise ama-mRNA ukuze aphathe izifo ezingeni lofuzo Indima ye.

▲ Inqubo yokuhlanganisa isuka ku-DNA iye ku-RNA iye kumaprotheni (Umthombo wesithombe: bing)

Njengamanje, izidakamizwa eziyinhloko ze-nucleic acid zifaka i-antisense nucleic acid (ASO), i-RNA encane ephazamisayo (siRNA), i-microRNA (miRNA), i-RNA encane eyenza i-RNA (saRNA), i-RNA yesithunywa (mRNA), i-aptamer, ne-ribozyme., Antibody nucleic acid conjugated drugs (ARC), njll.

Ngaphezu kwe-mRNA, ucwaningo nokuthuthukiswa kwezinye izidakamizwa ze-nucleic acid nakho sekuthole ukunakwa okwengeziwe eminyakeni yamuva.Ngo-2018, umuthi wokuqala emhlabeni we-siRNA (Patisiran) wagunyazwa, futhi kwaba umuthi wokuqala we-nucleic acid ukusebenzisa uhlelo lokulethwa kwe-LNP.Eminyakeni yamuva nje, ijubane lemakethe lezidakamizwa ze-nucleic acid nalo liye landa.Ngo-2018-2020 kuphela, kunezidakamizwa ezi-4 ze-siRNA, izidakamizwa ezintathu ze-ASO zivunyiwe (i-FDA ne-EMA).Ngaphezu kwalokho, i-Aptamer, i-miRNA neminye imikhakha nayo inemithi eminingi esesigabeni somtholampilo.

Izinzuzo nezinselele zezidakamizwa ze-nucleic acid

Kusukela ngeminyaka yawo-1980, ucwaningo nokuthuthukiswa kwemithi emisha esekelwe ekuqondisweni kuye kwanda kancane kancane, futhi kutholakale inani elikhulu lemithi emisha;izidakamizwa zendabuko zamakhemikhali ane-molecule encane kanye nezidakamizwa ezilwa namasosha omzimba zombili zinemiphumela yezemithi ngokubophezela ekuqondiseni amaprotheni.Amaprotheni okuhlosiwe angaba ama-Enzyme, ama-receptors, iziteshi ze-ion, njll.

Nakuba izidakamizwa ezine-molecule encane zinezinzuzo zokukhiqiza kalula, ukuphathwa ngomlomo, izakhiwo ezingcono ze-pharmacokinetic, kanye nokudlula kalula kulwelwesi lwamangqamuzana, ukuthuthukiswa kwazo kuthintwa ukugayeka kwezidakamizwa kulokho okuhlosiwe (nokuthi ingabe iphrotheni eqondiwe inokwakheka kwephakethe nosayizi ofanele)., Ukujula, i-polarity, njll.);ngokwe-athikili ku-Nature2018, angu-3,000 kuphela amaprotheni angama-20,000 afakwe ikhodi yofuzo lomuntu angaba imithi, futhi angama-700 kuphela anemithi ehambisanayo ethuthukisiwe (Ngokuyinhloko kuma-molecule amancane).

Inzuzo enkulu yezidakamizwa ze-nucleic acid ukuthi izidakamizwa ezahlukene zingathuthukiswa kuphela ngokushintsha ukulandelana kwesisekelo se-nucleic acid.Uma kuqhathaniswa nezidakamizwa ezisebenza ezingeni leprotheyini lendabuko, inqubo yokuthuthukiswa kwayo ilula, iyasebenza, futhi iqondene nebhayoloji;uma kuqhathaniswa nokwelashwa kwe-genomic DNA-level, izidakamizwa ze-nucleic acid azinayo ingozi yokuhlanganiswa kwezakhi zofuzo futhi ziguquguquka kakhulu ngesikhathi sokwelashwa.Umuthi ungayekwa uma kungekho ukwelashwa okudingekayo.

Izidakamizwa ze-Nucleic acid zinezinzuzo ezisobala njengokucaciswa okuphezulu, ukusebenza kahle okuphezulu kanye nomphumela wesikhathi eside.Kodwa-ke, ngezinzuzo eziningi kanye nentuthuko esheshayo, izidakamizwa ze-nucleic acid nazo zibhekene nezinselele ezihlukahlukene.

Okukodwa ukuguqulwa kwe-RNA ukuze kuthuthukiswe ukuzinza kwezidakamizwa ze-nucleic acid nokunciphisa ukuzivikela komzimba.

Okwesibili ukuthuthukiswa kwabathwali ukuqinisekisa ukuzinza kwe-RNA ngesikhathi senqubo yokudlulisa i-nucleic acid kanye nezidakamizwa ze-nucleic acid ukuze kufinyelelwe amaseli / izitho ezihlosiwe;

Okwesithathu ukuthuthukiswa kohlelo lokulethwa kwezidakamizwa.Indlela yokuthuthukisa uhlelo lokulethwa kwezidakamizwa ukuze kuzuzwe umphumela ofanayo ngemithamo ephansi.

Ukuguqulwa kwamakhemikhali kwezidakamizwa ze-nucleic acid

Izidakamizwa ze-nucleic acid zangaphandle zidinga ukunqoba izithiyo eziningi ukuze zingene emzimbeni ukuze zidlale indima.Lezi zithiyo ziye zadala nobunzima ekuthuthukisweni kwezidakamizwa ze-nucleic acid.Nokho, ngokuthuthukiswa kobuchwepheshe obusha, ezinye zezinkinga sezixazululiwe kakade ngokuguqulwa kwamakhemikhali.Futhi impumelelo yobuchwepheshe besistimu yokulethwa ibambe iqhaza elibalulekile ekwakhiweni kwezidakamizwa ze-nucleic acid.

Ukuguqulwa kwamakhemikhali kungathuthukisa ikhono lemithi ye-RNA yokumelana nokuwohloka kwama-endonuclease angapheli kanye nama-exonuclease, futhi kuthuthukise kakhulu ukusebenza kwezidakamizwa.Ngezidakamizwa ze-siRNA, ukuguqulwa kwamakhemikhali kungaphinda kuthuthukise ukukhethwa kwezintambo zabo ze-antisense ukuze kuncishiswe umsebenzi we-RNAi osuke uhlosiwe, futhi ushintshe izakhiwo ezingokomzimba namakhemikhali ukuze kuthuthukiswe amandla okulethwa.

1. Ukuguqulwa kwamakhemikhali kashukela

Esigabeni sokuqala sokuthuthukiswa kwezidakamizwa ze-nucleic acid, izinhlanganisela eziningi ze-nucleic acid zibonise umsebenzi omuhle we-biological in vitro, kodwa umsebenzi wabo ku-vivo wehla kakhulu noma walahleka ngokuphelele.Isizathu esiyinhloko ukuthi ama-nucleic acid angashintshiwe aphulwa kalula ngama-enzyme noma ezinye izinto ezingapheli emzimbeni.Ukuguqulwa kwamakhemikhali kashukela ikakhulukazi kuhlanganisa ukuguqulwa kwe-2-position hydroxyl (2'OH) kashukela ibe yi-methoxy (2'OMe), i-fluorine (F) noma (2'MOE).Lezi zinguquko zingakhuphula ngempumelelo umsebenzi nokukhetha, zinciphise imithelela engekho ekuqondisweni, futhi zinciphise imiphumela emibi.

▲ Ukuguqulwa kwamakhemikhali kashukela (umthombo wesithombe: inkomba 4)

2. Ukuguqulwa kwamathambo e-Phosphoric acid

Ukuguqulwa kwamakhemikhali okuvame ukusetshenziswa kakhulu komgogodla we-phosphate i-phosphorothioate, okungukuthi, umoya-mpilo onganqamuli kumgogodla we-phosphate we-nucleotide uthathelwa indawo sulphur (ukuguqulwa kwe-PS).Ukuguqulwa kwe-PS kungamelana nokuwohloka kwama-nuclease futhi kuthuthukise ukusebenzisana kwezidakamizwa ze-nucleic acid namaprotheni e-plasma.Umthamo wokubopha, wehlise izinga lokusulwa kwezinso futhi andise isigamu sempilo.

▲ Ukuguqulwa kwe-phosphorothioate (umthombo wesithombe: inkomba 4)

Nakuba i-PS ingase inciphise ukuhlobana kwama-nucleic acid kanye nezakhi zofuzo ezihlosiwe, ukuguqulwa kwe-PS kune-hydrophobic futhi kuzinzile, ngakho-ke kusewukuguqulwa okubalulekile ekuphazamiseni ama-nucleic acid amancane kanye ne-antisense nucleic acids.

3. Ukuguqulwa kwendandatho enamalungu amahlanu ye-ribose

Ukuguqulwa kwendandatho ye-ribose enamalungu amahlanu kubizwa ngokuthi ukuguqulwa kwamakhemikhali esizukulwane sesithathu, okuhlanganisa i-nucleic acid-locked nucleic acid BNAs, i-peptide nucleic acid PNA, i-phosphorodiamide morpholino oligonucleotide PMO, lezi zinguquko zingathuthukisa ngokwengeziwe izidakamizwa ze-nucleic acid Ukumelana ne-nuclease, njll.

4. Okunye ukuguqulwa kwamakhemikhali

Ekuphenduleni izidingo ezihlukene zezidakamizwa ze-nucleic acid, abacwaningi bavame ukwenza ukuguqulwa nokuguqulwa ezisekelweni namaketanga e-nucleotide ukwandisa ukuzinza kwezidakamizwa ze-nucleic acid.

Kuze kube manje, yonke imithi eqondiswe ku-RNA egunyazwe i-FDA iyi-analogue ye-RNA eyenziwe ngamakhemikhali, esekela ukusetshenziswa kokuguqulwa kwamakhemikhali.Ama-oligonucleotide anemigqa eyodwa yezigaba ezithile zokuguqulwa kwamakhemikhali ahluka kuphela ngokulandelana, kodwa wonke anezakhiwo ezifanayo zomzimba namakhemikhali, ngakho-ke anezindawo ezivamile ze-pharmacokinetics kanye ne-biological properties.

Ukulethwa nokuphathwa kwezidakamizwa ze-nucleic acid

Izidakamizwa ze-Nucleic acid ezithembele kuphela ekuguqulweni kwamakhemikhali zisawohloka kalula ngokushesha ekujikelezeni kwegazi, akulula ukunqwabelana ezicutshini ezihlosiwe, futhi akulula ukungena ngokuphumelelayo ulwelwesi lweseli oluhlosiwe ukuze lufinyelele indawo yesenzo ku-cytoplasm.Ngakho-ke, amandla ohlelo lokulethwa ayadingeka.

Njengamanje, ama-nucleic acid drug vectors ahlukaniswe ikakhulukazi ama-viral and non-viral vectors.Owokuqala uhlanganisa i-adenovirus-associated virus (AAV), i-lentivirus, i-adenovirus kanye ne-retrovirus, njll. Lokho kufaka phakathi abathwali be-lipid, ama-vesicle nokunye okunjalo.Ngokombono wezidakamizwa ezithengiswayo, ama-viral vectors nabathwali be-lipid bavuthwe kakhulu ekulethweni kwezidakamizwa ze-mRNA, kuyilapho izidakamizwa ezincane ze-nucleic acid zisebenzisa abathwali abaningi noma izinkundla zobuchwepheshe ezifana ne-liposomes noma i-GalNAc.

Kuze kube manje, izindlela zokwelapha eziningi ze-nucleotide, ezihlanganisa cishe zonke izidakamizwa ezivunyiwe ze-nucleic acid, seziye zasetshenziswa endaweni, njengamehlo, umgogodla nesibindi.Ama-Nucleotides ngokuvamile ama-hydrophilic polyanions amakhulu, futhi lesi sici sisho ukuthi azikwazi ukudlula kalula kulwelwesi lwe-plasma.Ngesikhathi esifanayo, izidakamizwa zokwelapha ezisekelwe kwi-oligonucleotide ngokuvamile azikwazi ukuwela umgoqo wegazi-ubuchopho (BBB), ngakho-ke ukulethwa ohlelweni lwezinzwa oluphakathi (CNS) kuyinselele elandelayo yezidakamizwa ze-nucleic acid.

Kuyaqapheleka ukuthi ukwakheka kokulandelana kwe-nucleic acid kanye nokuguqulwa kwe-nucleic acid kuyizinto okugxilwe kuzo abacwaningi kulo mkhakha.Ngokuguqulwa kwamakhemikhali, i-nucleic acid eshintshwe amakhemikhali, idizayini noma ukuthuthukiswa kokulandelana kwe-nucleic acid okungeyona yemvelo, ukwakheka kwe-nucleic acid, ukwakhiwa kwe-vector, izindlela ze-nucleic acid synthesis, njll. Izihloko zobuchwepheshe ngokuvamile ziyizifundo ezigunyazwe ilungelo lobunikazi.

Thatha i-coronavirus entsha njengesibonelo.Njengoba i-RNA yayo iyinto ekhona ngendlela yemvelo ngokwemvelo, “i-RNA ye-coronavirus entsha” ngokwayo ayinakunikwa ilungelo lobunikazi.Kodwa-ke, uma umcwaningi wesayensi ehlukanisa noma ekhipha i-RNA noma izingcezwana ezingaziwa kubuchwepheshe ezivela ku-coronavirus entsha okokuqala futhi ezisebenzisa (ngokwesibonelo, ukuyiguqula ibe umgomo), khona-ke kokubili i-nucleic acid nomgomo kunganikezwa amalungelo obunikazi ngokuvumelana nomthetho.Ngaphezu kwalokho, ama-molecule e-nucleic acid ahlanganiswe ngokwenziwa ocwaningweni lwe-coronavirus entsha, njengama-primer, ama-probes, i-sgRNA, ama-vectors, njll., zonke ziyizinto ezinelungelo lobunikazi.

Amazwi okuphetha

Ngokuhlukile kumshini wezidakamizwa zendabuko ze-molecule encane yamakhemikhali kanye nezidakamizwa ze-antibody, izidakamizwa ze-nucleic acid zinganweba ukutholakala kwezidakamizwa kuzinga lofuzo ngaphambi kwamaprotheni.Kuyaqapheleka ukuthi ngokwanda okuqhubekayo kwezinkomba kanye nokwenza ngcono okuqhubekayo kokulethwa nokuguqulwa kobuchwepheshe, izidakamizwa ze-nucleic acid zizosakaza iziguli eziningi zezifo futhi zibe elinye isigaba semikhiqizo yeziqhumane ngemva kwemithi yamakhemikhali e-molecule encane kanye nemithi elwa namasosha omzimba.

Izinto eziyisethenjwa:

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-frontotemporal/

3. Liu Xi, Sun Fang, Tao Qichang;Nhlakanipho Mphathi."Ukuhlaziywa kobunikazi bezidakamizwa ze-nucleic acid"

4. I-CICC: izidakamizwa ze-nucleic acid, isikhathi sesifikile

Imikhiqizo Ehlobene:

Ikhithi ye-Cell Direct RT-qPCR

Ikhithi ye-Mouse Tail Direct PCR

Ikhithi ye-PCR ye-Animal Tissue Direct