Intandaro ya buri kintu cyubuzima hamwe nubuvuzi bugezweho bwo kuvura bubeshya peptide. Iyi fondasiyo ya covalent ihuza ikiraro acide amino kugirango ibe molekile yibinyabuzima ikomeye. Kwimura ibisobanuro byibitabo byashize, kumenya neza imiti ihuza imiti byerekana neza ubucuruzi mubinyabuzima. Intungamubiri za poroteyine karemano zikora neza muri selile ya ribosome, zigera ku mikorere idasanzwe. Ibinyuranye, kwigana ibihimbano muburyo butangiza inzitizi zikomeye zubuhanga. Gutanga umusaruro, kubangamira isuku, no kunanirwa gutuza bikunze kwibasira laboratoire hamwe nimirongo yubucuruzi. Ugomba gusobanukirwa nukuri kwimiti yo gushiraho no gutandukanya kugirango usuzume neza uburyo bukoreshwa. Tuzareba uburyo gukomera kwimiterere bigira ingaruka zo gusesengura no gufata ingamba. Kurangiza, ubu bushishozi bufatika buzakuyobora muguhitamo abafatanyabikorwa bashoboye cyane kubikorwa peptide.
Imbonerahamwe
Rigidity Structural: Peptide bonds yerekana igice cyikubye kabiri, kigaragaza ituze hamwe na 3D ihinduka ryubuvuzi bwa sintetike hamwe na reagent.
Synthesis Bottlenecks: Imyitwarire ya reaction muri laboratoire (nka SPPS) isaba optimizasiyo ikomeye kugirango wirinde gukurikiranwa hamwe no guteranya.
Isesengura ry'imiterere: Kwemeza uburinganire busaba ibizamini bya chromatografique na spekrometrike (HPLC, MS) kugirango ubushakashatsi cyangwa isuku yo mu rwego rwa clinique.
Guhitamo Abafatanyabikorwa: Kuva mu bitekerezo ujya mu bucuruzi bisaba umufatanyabikorwa wa synthesis wasuzumwe ku bunini, QA / QC gukorera mu mucyo, n'ingamba zo kugabanya ingaruka.
Sisitemu y'ibinyabuzima yubaka poroteyine zigoye binyuze mu buryo bwuzuye bwo kubura umwuma. Itsinda rya carboxyl ya aside amine imwe ikora muriyi nzira. Ihuza itsinda rya amino ya aside amine yegeranye. Iyi miti yihariye irekura molekile imwe yamazi nkibicuruzwa. Ibisubizo bivamo covalent ihuza umugongo wibanze wa bose peptide . Ikora nkibanze shingiro ryimiterere ifata ubuzima hamwe. Twise iyi sano yihariye ihuza amide. Abashinzwe imiti bize imiterere yihariye mumyaka mirongo kugirango bumve imiterere yibiyobyabwenge.
Inkunga ya karubone-azote ntabwo ari umugozi woroshye. Electron ikomeza guhora hejuru ya ogisijeni, karubone, na azote. Kugabana electron ikora imiterere ya resonance itandukanye. Inkunga yunguka igice cyikubye kabiri. Ntishobora kuzunguruka mu kirere. Atome esheshatu zirimo zirimo kuguma zifunze burundu muri geometrie ikomeye. Iboneza rya Trans irashimwa cyane mubisanzwe. Ibi birinda amakimbirane akomeye hagati yiminyururu nini ituranye. Proline yerekana ibintu bidasanzwe aho cis iboneka. Gufunga geometrike bigabanya cyane molekulike ihinduka.
Ni ukubera iki uku gukomera gukomeye bifite akamaro kanini? Ihindura mu buryo butaziguye igishushanyo mbonera no gutsinda kwa sintetike yo kuvura. Kubura kuzunguruka bishyira mubikorwa byihariye-bitatu. Iyi miterere iteganijwe igena selile yakira ihuza isano. Gukomera kandi birinda umugongo wa molekuline kwangirika kwimisemburo yihuse. Yagura ibikorwa byubuzima bwa farumasi yubucuruzi. Iyo abashakashatsi bashushanya ibihimbano, bashingira cyane kuri geometrie iteganijwe. Imiterere ya planar itegeka muri rusange muri vivo bioavailability. Niba inkwano yari ihindagurika rwose, ibiyobyabwenge bigamije kunanirwa kwifungisha neza.
Ibinyabuzima biranga ibinyabuzima |
Umutungo wumubiri |
Ibisubizo byubucuruzi |
|---|---|---|
Delonisation |
Igice cya kabiri-gihuza imiterere |
Irinda umugongo kuzunguruka, ushimangira imiterere ihamye. |
Umubumbe wa Geometrie |
Ibikoresho byahinduwe neza |
Kugabanya impande zumunyururu zishyamiranye, kunoza reseptor. |
Guhindura umwuma |
Covalent umugongo uhuza |
Irasaba ingufu zihariye zinjiza, bigoye gukora synthique-up. |
Ubusobanuro bwibinyabuzima bukoresha selile ya ribosome neza. Iyi mashini yibinyabuzima ikora neza cyane kandi igenzurwa cyane. Ihuza hamwe amino acide bitagoranye mubidukikije byamazi. Sintezike yimiti ikora muburyo butandukanye muri laboratoire. Synthesis ikomeye-Phase Peptide (SPPS) ikora nkibipimo byubucuruzi bigezweho. Irasaba guhuza ibinyabuzima bigoye nka HATU cyangwa DIC. Tugomba kurinda amatsinda yihariye akora. Amatsinda asanzwe arengera arimo Fmoc cyangwa Boc chemistries. Noneho, duhitamo kubatesha agaciro dukoresheje ibice byoroheje cyangwa acide. Uru ruzinduko rukaze rusubiramo kuri buri aside amine yongeyeho. Imiterere yimiti itegeka kugenzura ibidukikije bidasanzwe. Tugomba guhatira reaction ya biologiya ikora muburyo busanzwe.
Intege nke zirahari nubwo kamere gakondo ikomeye ya covalent. Cleavage, akenshi itwarwa na hydrolysis, ikomeza guhungabanya inganda. Ibidukikije bimwe byihutisha iyangirika ryimiti byihuse. Ugomba guteganya inzira zisenyuka mugihe cyo gutegura.
Ibidukikije bikabije bya pH bihungabanya ingufu za electrostatike iringaniye, itera hydrolysis yihuse.
Imisemburo ya proteolyique yibasira cyane kandi igabanya moteri yihariye ya aside amine.
Guhangayikishwa cyane nubushyuhe butangiza imbaraga za kinetic zirenze, guca umugongo uhuza imburagihe.
Kwanduza mikorobe kwinjizamo ibikorwa byimikorere idahwitse muburyo butemewe.
Ubushuhe bukabije bw’ibidukikije butera kwangirika kwa hydrolytike mu ifu ya lyofilize ifunze neza.
Gusuzuma imipaka ntarengwa bisaba ibintu bifatika. Ntushobora gukora ibicuruzwa byubucuruzi buhumyi. Ingaruka za Cleavage zitegeka protocole ikomeye yo kubika kwisi yose. Kurugero, ifu ya lyofilize isaba ubushyuhe buri munsi ya zeru. Amazi yo mumazi arasaba ibintu byihariye byoherejwe. Ibikoresho byo gupakira bigomba guhagarika byimazeyo nubushyuhe bwa UV. Izi mbogamizi zikomeye zigira ingaruka zitaziguye mubucuruzi no kohereza ibikoresho. Niba umugongo wumugongo ucitse imburagihe, ubuvuzi bwose butakaza imikorere. Inzego zishinzwe kugenzura nka FDA zisaba amakuru yuzuye, imyaka myinshi ihamye. Ugomba kwerekana umugongo ukomeza kuba mwiza mugihe runaka.
Kugenzura ubuziranenge byerekana ikibazo gikomeye cyo gukora molekile. Ugomba kwerekana ubunyangamugayo muburyo bwuzuye mbere yo gusaba. Gutandukanya intego ya molekile na synthesis byproducts birazwi ko bigoye. Urutonde rwaciwe rugaragaza neza ibicuruzwa byanyuma bifuza kumubiri. Gusiba umwanda uhisha byoroshye mumvange ivanze. Ihuriro rimwe ryabuze ryangiza urukurikirane rwose. Ukeneye uburyo bukomeye, bwihariye bwo gusesengura kugirango umenye intsinzi.
Uburyo bwo gusesengura |
Igikorwa Cyibanze |
Intego yo Kwemeza Intego |
Imiterere y'ibisohoka |
|---|---|---|---|
HPLC |
Gutandukana kwa Chromatografiya |
Muri rusange urutonde rwubuziranenge |
Impinga ya chromatogramu |
Mass Spectrometry |
Ibipimo bya Ion |
Uburemere bwa molekile |
Igishushanyo mbonera |
NMR Spectroscopy |
Magnetic Resonance Imaging |
Icyemezo cya kabiri cyemeza |
Guhindura imiti |
FTIR Spectroscopy |
Imirasire yumucyo |
Hagati aho inkwano ihari |
Absorption impinga |
Amazi meza ya Chromatografiya (HPLC) akomeza kuba igipimo cyizahabu rusange. Itandukanya ibice bishingiye rwose kuri hydrophobicity. HPLC itandukanya intego yuzuye kuva kumyanda migufi yaciwe. Iraguha ijanisha risobanutse, rifite intego. Mass Spectrometry (MS) igenzura uburemere bwa molekile itaziguye. ESI na MALDI-TOF byerekana tekinike zikoreshwa cyane. MS itanga ibyemezo byingenzi bikurikiranye ako kanya. Iremeza aside amine yose ihujwe neza mugihe cya synthesis. Irerekana utuntu duto duto duterwa no kubura amatsinda arinda byoroshye.
Nuclear Magnetic Resonance (NMR) na Fourier Transform Infrared Spectroscopy (FTIR) byemeza ibintu byimbitse. Bemeza neza amide ihari neza. Bafasha abashakashatsi gushushanya ibyiciro bya kabiri neza. Ibi bikoresho byerekana alpha-helices cyangwa beta-impapuro zikora mumurongo. Ibisobanuro byamakuru birakenewe rwose kubaguzi babikoresho. Mugihe usubiramo icyemezo cyuwabikoze (CoA), reba neza. Saba neza neza chromatogramu ya HPLC yerekana impinga zikarishye. Menya neza ko amakuru ya MS ahuye na theoretique yabazwe neza. Vague CoAs yerekana ko hashobora kubaho inenge cyangwa umwanda uhishe. Ukeneye ibimenyetso bifatika byerekana ubuziranenge bwibicuruzwa mbere yo gukomeza.
Kwiyongera k'umusaruro wimiti utangiza laboratoire zikomeye kwisi yose. Umusaruro ugabanuka cyane muburyo bukurikiranye bidasanzwe. Kunanirwa kunanirwa byiyongera hamwe na aside aside yongeyeho. Imibare yukuri ya synthesis ikurikirana ntabwo ibabarira.
99% guhuza neza kuri buri ntambwe bitanga hafi 60% byera hejuru yintambwe 50.
95% imikorere igabanuka muri rusange umusaruro ukurikirana kuri 7% gusa.
Iminyururu yananiranye ihinduka imiti itandukanijwe na misa ya nyuma.
Igiciro cyo kweza cyapimye cyane uko uburebure bwikurikiranya bwiyongera burenze 30.
Inzitizi ya Steric irusheho kugora imbaraga nini zo guhuza imbaraga. Urunigi runini rwuruzitiro ruhagarika ibikorwa bikora. Guteranya bibaho kenshi muburyo bwa hydrophobique. Urunigi rwa chimique rukura rugenda rwisubiraho. Ukuzenguruka guhisha rwose itsinda rya amino rikora. Igipimo gisanzwe cyo guhuza ntigishobora kwinjira mubwinshi bwuzuye. Kugira ngo ibyo bishoboke, abahanga mu bya shimi bakoresha imiti yihariye. Bashyiraho ibintu bikomeye bya chaotropique kugirango bahungabanye hydrogene. Rimwe na rimwe, bashyiramo ubushyuhe bwa microwave kugirango bahuze iminyururu itandukanye. Abashakashatsi bakunze gusimbuza pseudo-proline kugirango birinde uku kuzura imburagihe.
Igipimo nacyo gihatira ibyemezo byubukungu bigoye, bifatika. Uhura nigiciro gihoraho hamwe nubucuruzi bwera buri munsi. Kugera kuri ultra-high 99% isuku bisaba kwagura HPLC. Ubu buryo bwihariye butwara ibintu byinshi bihenze, byangiza. Iratanga kandi umusaruro wibikoresho byingenzi mugihe cyo gutandukana. Ivuriro muri vivo gusaba risaba uku kwezwa gukomeye nta shiti. Ibinyuranye, ibyingenzi mubushakashatsi bwa vitro birashobora kwihanganira 85% byera neza. Ugomba guhuza byimazeyo ibikorwa byimari nibisabwa byubushakashatsi. Guhitamo urwego rutari ruto rwangiza ingengo yimari kandi ikabangamira amakuru akomeye.
Kwimuka ukava mubitekerezo byerekeranye nubucuruzi bwuzuye bisaba ubufatanye bufatika. Ugomba gusuzuma ubushobozi bwa tekiniki mbere yo gusinya amasezerano. Umucuruzi watoranijwe arashobora gukemura ibibazo bidasanzwe? Shakisha uburambe bwagenzuwe hamwe na aside amine idasanzwe. Reba ubushobozi bwabo bwumubiri bwo guhuza imiterere yikurikiranya. Menya niba bakoresha uburyo bukomeye cyangwa ibyiciro byamazi. LPPS ikora neza mugihe gito cyane, cyinshi-cyubucuruzi bukurikirana. SPPS ikora igihe kirekire, igoye cyane yubushakashatsi. Umucuruzi agomba guhuza tekinoroji yabo na molekile yawe yihariye.
Ubwishingizi bufite ireme butandukanya abafatanyabikorwa bazwi na laboratoire yibanze. Inganda-yubushakashatsi iratandukanye cyane no kubahiriza cGMP. Ibikorwa byiza byo gukora muri iki gihe bisaba kugenzura ibidukikije bikaze. Umucuruzi agomba gukomeza inyandiko zisanzwe zikoreshwa (SOPs) ubudahwema. Bakeneye ubwiherero bwabigenewe, bukurikiranwa cyane kugirango babone umusaruro. Kugenzura ubuziranenge bw’ikirere bigomba gukomeza ubudahwema. Ugomba gusaba raporo yubugenzuzi burambuye kubipimo byubahiriza ibikoresho.
Gukorera mu mucyo ntabwo rwose biganirwaho kubaguzi bakomeye. Shiraho ibyateganijwe gutegurwa hakiri kare mubiganiro. Saba raporo yisesengura yatanzwe hamwe na buri cyiciro. Kugenzura neza HPLC hamwe na MS spectra ni itegeko rwose. Baza amakuru yuzuye yemeza amakuru imbere. Niba umucuruzi atinya gusangira amakuru yisesengura mbisi, genda uhite. Mubisanzwe byerekana kunanirwa kugenzura ubuziranenge.
Suzuma ingamba zo kugabanya ingaruka nibihe byahindutse. Kunanirwa kwa Synthesis bibaho kenshi muri chimie igoye. Umufatanyabikorwa mwiza amenyesha ibyo bibazo bya tekiniki ako kanya. Batezimbere intambwe igoye yo guhuza ibikorwa batabigusabye. Bahindura protocole yimbere kugirango bubahirize igihe ntarengwa cyumushinga. Ibyo babigeraho batigeze batamba ibicuruzwa byanyuma. Amateka akomeye y'abacuruzi arahanura neza umusaruro wizewe. Ukeneye umufatanyabikorwa ushyikirana cyane uyobora inzitizi zubuhanga.
Kumenya chimie yibanze ya peptide ni intangiriro gusa. Guhindura chimie yoroheje mubikorwa byizewe, binini byerekana ikibazo cyubucuruzi. Twasuzumye uburyo gukomera kwimiterere gutegeka gutuza no gukora biologiya. Twasuzumye impamvu synthesis ikurikirana itangiza umusaruro wimibare idashobora kwirindwa.
Igenzura rikomeye ryisesengura rikomeje kuba imbaraga zawe zikomeye zo kwirinda kwangirika no gutsindwa kwa synthesis. Ubufatanye bw'abacuruzi bugamije kugabanya ingaruka zikomeye ziterwa no gupima imiti. Nkumuguzi wa tekiniki cyangwa umushakashatsi, ugomba gusaba gukorera mu mucyo. Kugenzura protocole yawe ya none irwanya ibipimo bigezweho byo gusesengura. Saba inama zijyanye na tekiniki kuburyo bukurikiranye burenze ibisigazwa mirongo itatu. Ongera usuzume icyitegererezo CoAs mbere yo kwiyemeza amasezerano manini yo gukora ibicuruzwa.
Igisubizo: Mugihe imigozi yose ya peptide iba muburyo bwa tekiniki hagati yububiko, ibaho muburyo bwimiterere ya biologiya ya alfa-amino acide. Bahuza itsinda rya alpha-carboxyl ya aside amine imwe nitsinda rya alpha-amino yandi. Iyi gahunda yihariye ya biologiya itera resonance idasanzwe ihamye, igahatira geometrie ikomeye, planari ya ngombwa kugirango ifumbire ya poroteyine.
Igisubizo: Hydrolysis nintandaro yambere yo kwangirika kwimiterere. Ubushuhe bukora nk'umusemburo, uca covalent ihuza. Iyi myitwarire yihuta cyane mugihe ihindagurika rikabije rya pH, guhura nubushyuhe bwinshi, cyangwa mugihe kwanduza mikorobe kubwimpanuka byinjiza imisemburo ya proteolyique idateganijwe mububiko.
Igisubizo: Synthesis isaba kwiyongera gukurikiranye. Niba buri ntambwe yo guhuza ikora kuri 99% ikora neza, igipimo cya 1% cyatsinzwe kibara imibare. Intambwe zirenga 50 zikurikiranye, umusaruro rusange uragabanuka kugera kuri 60%. Inzitizi ya Steric hamwe no gukusanya urunigi bikomeza guhagarika imbuga zikora, kwemeza guhuza kwuzuye kutuzuye nkuko urunigi rurambura.
Igisubizo: Ugomba gusaba Icyemezo cyisesengura kirimo raporo zisesenguye. Saba chromatogramu ya HPLC ihanitse kugirango yerekane ubuziranenge bwicyitegererezo no kumenya umwanda waciwe. Icyarimwe, saba amakuru ya Mass Spectrometry (MS) kugirango umenye uburemere nyabwo bwa molekile, urebe neza niba bikurikiranye kandi bikingire gukuraho amatsinda.
