Ukuqonda izixhobo zokuqalisa ichiza le-polypeptide

Amachiza e-peptide achazwa ngokubanzi njengeepolima ezenziwe ngamabhondi e-amide kunye neentsalela ze-amino acid ezingaphantsi kwama-40.Ngenxa yomsebenzi ophezulu we-receptor kunye nokukhetha kwamachiza e-peptide kunye nomngcipheko ophantsi wemiphumo emibi, kuye kwaba nomdla oqinileyo kwiipeptides ezivela kumashishini amayeza.Ngeli xesha, bekukho amachiza amaninzi eenkwenkwezi, awayegxile kakhulu kwishishini lesifo se-metabolic, njenge-GLP-1 analogue somalitide, gastric inhibitory peptide (GIP) glucagon-like peptide-1 (GLP-1) tesiparatide kunye nezinye ezimbini. -i-receptor agonists.Ukongeza, ngokunyuka kwamachiza ePDC kunye neRDC.Okwangoku, iindlela zokulungiselela iziyobisi ze-polypeptide zibandakanya i-chemical synthesis kunye ne-biological fermentation.I-Biofermentation isetyenziselwa ukuvelisa iipeptides ezinde.Iinzuzo ziindleko eziphantsi zokuvelisa, kodwa ukungakwazi ukwazisa i-amino acids engeyondalo kulandelelwano lwe-peptide kunye nokungakwazi ukwenza imihombiso eyahlukeneyo kwikhonkco le-peptide.Ngoko ke, ukusetyenziswa kwayo kuncinci kakhulu.Iindlela zokudityaniswa kweekhemikhali ziquka ukuhlanganiswa kwesigaba esiqinileyo kunye nesigaba solwelo.I-Solid-phase synthesis ine-advanteji ebalulekileyo ngaphezu kwe-liquid-phase synthesis: ubuninzi bezinto eziphathekayo zingasetyenziselwa ukusabela ukuqinisekisa ukudibanisa okupheleleyo.Ukugqithiswa kwe-amino acids, i-shrinkage agents, kunye ne-products inokususwa ngemisebenzi yokucoca elula, ukuphepha umsebenzi onzima emva kokulungiswa kunye nokuhlanjululwa kunye nokuphucula ukusebenza kakuhle, ngoko ke indlela yokudibanisa isigaba esiqinileyo iye yasetyenziswa kakhulu."Imathiriyeli ekrwada yokwenziwa kweekhemikhali zokwenziwa kweepeptides ibandakanya izixhobo zokuqalisa, ii-reagents, kunye nezinyibilikisi."Umgangatho wabo, ngokukodwa umgangatho wezinto zokuqala, unokuba nempembelelo eyahlukileyo kumgangatho we-API.Izinto zokuqala zibhekisela kwizinto eziqinisekisiweyo ze-amino acid derivatives ze-peptide chain modified fatty acids, i-polyethylene glycol, njl. Njengeziqhekeza ezibalulekileyo zesakhiwo, zihlelwa njengezinto eziphathekayo kwisakhiwo se-API, ehambelana ngokuthe ngqo nomgangatho we-API.Ngoko ke, kufuneka sigxininise ekulawuleni izinto zokuqala.

I. Lungiselela ukhetho lokuqala lwemathiriyeli

I-ICHQ11 iphakamisa ngokucacileyo ukuba ukuba imveliso yekhemikhali ethengiswa kwimarike isetyenziswa njengemathiriyeli yokuqala ekrwada, umfaki-sicelo akadli ngokudinga ukuxoxa ngengqiqo yayo.Iimveliso zemichiza ezithengiswa kwimarike ngokubanzi azinakusetyenziselwa kuphela njengezixhobo zokuqala zamachiza, kodwa zinokuthengiswa nakwiimarike ezingezizo ezamayeza.Iikhompawundi ezenziwe ngokwezifiso kunye nezidityanisiweyo azikho kwiimveliso zemichiza ezithengiswa kwimarike.Nangona ingekho imakethi engeyoyamayeza ukukhusela i-amino acids ukuhlangabezana nenkcazo ye-ICHQ11 yeekhemikhali ezithengiswa kwimarike, zihlangene, zihluke ngokwekhemikhali kwaye zicacile ngokwesakhiwo, kulula ukuzihlukanisa kunye nokuhlanjululwa, kwaye zinokuchongwa kwaye zivavanywe ngeendlela eziqhelekileyo zohlalutyo. .Zineempawu ezizinzileyo zeekhemikhali kwaye kulula ukuzigcina, ukuthutha, kunye nokudibanisa

Ii.Ukulawulwa kwezinto ezichaphazelekayo kwizinto zokuqala

I-amino acids ekhuselweyo ekhankanywe ngasentla ifakwe kwisakhiwo se-API njengenxalenye ebalulekileyo yesakhiwo, ehambelana ngokuthe ngqo nomgangatho we-API.Ngoko ke, kufuneka silawule ngokungqongqo umxholo wokungcola kwizinto zokuqala, siqonde ukuguqulwa kunye nokususwa kobu kungcola kwinkqubo emiselweyo, kwaye ekugqibeleni sicacise ubudlelwane phakathi kwabo kunye nokungcola kwi-API.

Ukuqonda izixhobo zokuqalisa ichiza le-polypeptide

Okwesithathu, intsalela ye-solvent kwizinto zokuqala

Ngokubanzi, ukunikezelwa ngokukodwa kwesigaba esiqinileyo sokuvelisa i-peptides, isixa esikhulu se-solvent siya kusetyenziswa ukucoca i-peptide resin emva kokugqiba inyathelo ngalinye lokudibanisa i-amino acid kunye ne-detachment ekukhuseleni.Iipeptide ezikrwada ezifunyenwe ngokukrazula intlaka yepeptide nazo ziya kwenziwa yiHPLC kwaye zomiswe ngumkhenkce.Ngaloo ndlela, kukho umngcipheko omncinci wokuba umlinganiselo omncinci we-solvent efakwe kwi-amino acids ekhuselayo iya kuhanjiswa kwi-API yokugqibela.Nangona kunjalo, ingqalelo ekhethekileyo kufuneka ihlawulwe kwiintsalela ze-acetate, i-acetate ye-butyl, kunye ne-solvents yotywala, njengoko ezi zinyibilikisiweyo zingabangela iziphumo ezichaphazelekayo kunye ne-amino acids esebenzayo okanye i-peptide chain ngexesha lokudibanisa okusebenzayo kwe-amino acids.Ngokomzekelo, ngexesha lokudibanisa i-amino acid, i-acetic acid eseleyo iya kusabela kunye neqela le-amino eliveziweyo kwikhonkco le-peptide, okukhokelela ekupheleni kokuvalwa kwe-peptide chain;Ngexesha lomsebenzi we-amino acid, isinyibilikisi sotywala esishiyekileyo sinokusabela kunye neqela le-carboxyl elisebenzayo, elikhokelela ekugqithiseni i-amino acid esebenzayo, ukunciphisa ukulingana kwe-amino acid, kwaye ekugqibeleni kubangele ukudityaniswa kwe-amino acid engaphelelanga kunye nokungabikho kokungcola kwe-peptide.Inkampani ilawula i-butyl acetate, i-alcohol, i-methanol, kunye ne-acetic acid kwi-COA, ithatha i-amino acid evela kuZheng Yuan Biochemical njengomzekelo.Umgangatho we-acetate ye-butyl yayingu-≤0.5% i-acetate ye-butyl, efunyenwe ngokwenene ukuba yi-0.10%.Ngokutsho kwe-ICHQ3C, i-butyl acetate kwiintlobo ezintathu ze-solvents, ibeka umgangatho we-0.5% okanye ngaphantsi ngokuhambelana neemfuno ze-ICHQ3C, kodwa ukuqwalasela i-butyl acetate amino acetylation kunokukhokelela kumngcipheko, kwakhona ukujongana ne-butyl acetate ukulinganisa uphando. , ukumisela imigangatho efanelekileyo ngakumbi.