Iipeptides luhlobo lweekhompawundi ezenziwe ngokudityaniswa kwee-amino acids ezininzi ngeebhondi zepeptide.Zikho kuyo yonke indawo kwizinto eziphilayo.Ukuza kuthi ga ngoku, amashumi amawaka eepeptides afunyenwe kwizinto eziphilayo.I-Peptides idlala indima ebalulekileyo ekulawuleni imisebenzi esebenzayo yeenkqubo ezahlukeneyo, izitho, izicubu kunye neeseli kunye nemisebenzi yobomi, kwaye zihlala zisetyenziselwa uhlalutyo olusebenzayo, uphando lwe-antibody, ukuphuhliswa kweziyobisi kunye nezinye iinkalo.Ngophuhliso lwetekhnoloji yebhayoloji kunye netekhnoloji ye-peptide synthesis, ayanda amayeza eepeptide aphuhlisiwe kwaye asetyenziswa eklinikhi.
Kukho iintlobo ezininzi zokuguqulwa kwe-peptide, ezinokuthi zihlulwe ngokulula zibe yi-post modification kunye nokuguqulwa kwenkqubo (usebenzisa ukuguqulwa kwe-amino acid ephumayo), kunye nokuguqulwa kwe-N-terminal, ukuguqulwa kwe-C-terminal, ukuguqulwa kwecala, ukuguqulwa kwe-amino acid, ukuguqulwa kwamathambo, njl., kuxhomekeke kwindawo yokuguqula (Umfanekiso 1).Njengendlela ebalulekileyo yokutshintsha isakhiwo sekhonkco eliphambili okanye amaqela ekhonkco ecala lamatyathanga e-peptide, ukuguqulwa kwepeptide kunokutshintsha ngokufanelekileyo iimpawu zomzimba kunye neekhemikhali zeekhompawundi ze-peptide, ukwandisa ukunyibilika kwamanzi, ukwandisa ixesha lesenzo kwi-vivo, ukutshintsha ukusabalalisa kwabo kwebhayoloji, ukuphelisa i-immunogenicity. , ukunciphisa iziphumo ebezingalindelekanga ezinetyhefu, njl. Kweli phepha, izicwangciso ezininzi zokuguqula iipeptide kunye neempawu zazo ziyaziswa.
1. Ukuhamba ngebhayisekile
Iipeptide ze-cyclic zinezicelo ezininzi kwi-biomedicine, kwaye iipeptide ezininzi zendalo ezinomsebenzi webhayoloji ziipeptides zebhayoloji.Ngenxa yokuba i-cyclic peptides ithande ukuba ngqongqo ngakumbi kune-peptides enomda, ixhathisa kakhulu kwinkqubo yokugaya, inokuphila kwindlela yokugaya, kwaye ibonise ubudlelwane obunamandla kwii-receptors ekujoliswe kuzo.Ukuhamba ngebhayisikile yeyona ndlela ingqalileyo yokwenza i-cyclic peptides, ngakumbi kwiipeptides ezinamahlakala amakhulu.Ngokwendlela ye-cyclization, inokwahlulwa ibe yi-side chain-side chain type, i-terminal-side chain type, i-terminal - uhlobo lwe-terminal (isiphelo ukuya kuhlobo lokuphela).
(1) sidechain-to-sidechain
Olona hlobo luqhelekileyo lwe-side-chain ukuya kwi-side-chain cyclization yi-disulfide bridging phakathi kweentsalela ze-cysteine.Le cyclization iqaliswa ngepere yeentsalela ze-cysteine ezikhutshiweyo kwaye emva koko zifakwe i-oxidized ukwenza iibhondi ze-disulfide.I-Polycyclic synthesis inokufezekiswa ngokususa okukhethiweyo kwamaqela okukhusela i-sulfhydryl.I-Cyclization ingenziwa okanye kwi-solvent-post-dissociation okanye kwi-resin yangaphambili ye-dissociation.Ukuhamba ngebhayisikile kwiireyini kunokungasebenzi kancinci kune-solvent cyclization kuba iipeptides kwiiresin azenzeki lula iiconformations zecyclified.Olunye uhlobo lwe-side-chain chain cyclization kukwenziwa kwesakhiwo se-amide phakathi kwe-aspartic acid okanye i-glutamic acid residue kunye nesiseko se-amino acid, efuna ukuba iqela lokukhusela ikhonkco elisecaleni kufuneka likwazi ukususwa ngokukhethiweyo kwi-polypeptide nokuba kwi-resin okanye emva kokuqhawula.Uhlobo lwesithathu lwe-side chain - i-side chain cyclization kukubunjwa kwe-diphenyl ethers nge-tyrosine okanye i-p-hydroxyphenylglycine.Olu hlobo lokuhamba ngebhayisikile kwiimveliso zendalo lufumaneka kuphela kwiimveliso ezincinci, kwaye iimveliso zebhayisikile zihlala zinexabiso elinokubakho lamayeza.Ukulungiswa kwezi khompawundi kufuna iimeko zokusabela ezizodwa, ngoko ke azisoloko zisetyenziswa kwi-synthesis yeepeptides eziqhelekileyo.
(2) iterminal-to-sidechain
I-Terminal-side chain cyclization idla ngokubandakanya i-C-terminal kunye neqela le-amino le-lysine okanye i-ornithine ye-side chain, okanye i-N-terminal ene-aspartic acid okanye i-glutamic side chain.Enye i-polypeptide cyclization yenziwa ngokwenza iibhondi ze-ether phakathi kwetheminali C kunye ne-serine okanye i-threonine side chain.
(3) Uhlobo lwesiphelo okanye intloko-to-msila
Iipolypeptides zamatsheyini zinokujikeleziswa kwisinyibilikisi okanye zilungiswe kwireyini ngecala lokujikeleza kwekhonkco.Uxinzelelo oluphantsi lweepeptides kufuneka lusetyenziswe kwi-solvent centralization ukuphepha i-oligomerization yeepeptides.Isivuno se-head-to-tail synthetic ring polypeptide sixhomekeke kulandelelwano lwe-polypeptide yekhonkco.Ke ngoko, ngaphambi kokulungiselela iipeptide ze-cyclic kwinqanaba elikhulu, ithala leencwadi leepeptide ezikhokelela ekubophelelekeni kufuneka lenziwe kuqala, lilandelwe yibhayisekile ukufumana ulandelelwano ngezona ziphumo zibalaseleyo.
2. I-N-methylation
I-N-methylation iyenzeka kuqala kwiipeptide zendalo kwaye ifakwe kwi-peptide synthesis ukuthintela ukubunjwa kwe-hydrogen bond, ngaloo ndlela yenza iipeptides zixhathise ngakumbi kwi-biodegradation kunye nokukhutshwa.Ukwenziwa kweepeptides kusetyenziswa iN-methylated amino acid derivatives yeyona ndlela ibalulekileyo.Ukongeza, i-Mitsunobu reaction ye-N-(2-nitrobenzene sulfonyl chloride) i-polypeptide-resin intermediates ene-methanol nayo ingasetyenziswa.Le ndlela isetyenziselwe ukulungiselela iilayibrari ze-cyclic peptide ezine-N-methylated amino acids.
3. Phosphorylation
I-Phosphorylation yenye yezona zinto ziqhelekileyo emva kokuguqulwa kwenguqu kwindalo.Kwiiseli zomntu, ngaphezu kwe-30% yeeprotheni zine-phosphorylated.I-phosphorylation, ngakumbi i-phosphorylation eguqulwayo, idlala indima ebalulekileyo ekulawuleni iinkqubo ezininzi zeselula, ezifana nokudluliselwa kwesignali, i-gene expression, i-cell cycle kunye ne-cytoskeleton regulation, kunye ne-apoptosis.
I-phosphorylation inokubonwa kwiindidi ze-amino acid eziseleyo, kodwa iithagethi eziqhelekileyo ze-phosphorylation ziyi-serine, i-threonine, kunye neentsalela ze-tyrosine.I-Phosphotyrosine, i-phosphothreonine, kunye ne-phosphoserine derivatives zinokungeniswa kwiipeptides ngexesha lokudibanisa okanye zenziwe emva kwe-peptide synthesis.I-phosphorylation ekhethiweyo inokufezekiswa ngokusebenzisa iintsalela ze-serine, i-threonine, kunye ne-tyrosine esusa ngokukhethiweyo amaqela okukhusela.Ezinye ii-reagents ze-phosphorylation zinokuzisa amaqela e-asidi ye-phosphoric kwi-polypeptide ngokuguqulwa kweposi.Kwiminyaka yakutshanje, i-phosphorylation ye-site-specific ye-lysine ifunyenwe ngokusebenzisa i-chemically selective Staudinger-phosphite reaction (Figure 3).
4. Myristoylation kunye palmitoylation
I-Acylation ye-N-terminal ene-fatty acids ivumela i-peptides okanye iiprotheni ukuba zibophe kwiimbrane zeseli.Ulandelelwano lwe-myridamoylated kwi-N-terminal yenza ukuba i-Src family protein kinases kunye ne-reverse transcriptase Gaq iiprotheyini zijoliswe ekubopheni kwiimbumba zeseli.I-asidi ye-Myristic yayidityaniswe ne-N-terminal ye-resin-polypeptide isebenzisa iimpendulo eziqhelekileyo zokudibanisa, kunye nesiphumo se-lipopeptide sinokudityaniswa phantsi kweemeko eziqhelekileyo kwaye zihlanjululwe yi-RP-HPLC.
5. Glycosylation
I-Glycopeptides efana ne-vancomycin kunye ne-teicolanin zi-antibiotics ezibalulekileyo kunyango lwe-bacterial-resistant infections, kunye nezinye ii-glycopeptides zihlala zisetyenziselwa ukuvuselela amajoni omzimba.Ukongeza, kuba ii-antigens ezininzi ze-microbial zine-glycosylated, kubaluleke kakhulu ukufunda i-glycopeptides ukuphucula isiphumo sonyango losulelo.Kwelinye icala, kuye kwafunyaniswa ukuba iiproteni ezikwinwebu yeseli yeeseli zethumba zibonisa i-glycosylation engaqhelekanga, eyenza ukuba i-glycopeptides idlale indima ebalulekileyo kumhlaza kunye nophando lokhuselo lomzimba.IiGlycopeptides zilungiswa ngeFmoc/t-Bu indlela.Iintsalela ze-glycosylated, ezifana ne-threonine kunye ne-serine, zihlala zingeniswa kwi-polypeptides nge-pentafluorophenol ester esebenzayo ye-fMOCs ukukhusela i-glycosylated amino acids.
6. I-isoprene
I-Isopentadienylation iyenzeka kwiintsalela ze-cysteine kwi-chain chain kufuphi ne-C-terminal.Iprotheyini ye-isoprene inokuphucula ubudlelwane be-membrane yeseli kwaye yenze i-protein-protein interaction.Iiprotheyini ze-Isopentadienate ziquka i-tyrosine phosphatase, i-GTase encinci, iimolekyuli ze-cochaperone, i-nuclear lamina, kunye neeprotheni ezibophelelayo ze-centromeric.I-Isoprene polypeptides inokulungiswa ngokusebenzisa i-isoprene kwii-resins okanye ngokuzisa i-cysteine derivatives.
7. Polyethylene glycol (PEG) ukuguqulwa
Ukuguqulwa kwe-PEG kungasetyenziselwa ukuphucula iprotheyini ye-hydrolytic stability, biodistribution kunye ne-peptide solubility.Ukuqaliswa kwamakhonkco e-PEG kwiipeptides kunokuphucula iimpawu zabo ze-pharmacological kunye nokuthintela i-hydrolysis ye-peptides nge-proteolytic enzymes.I-PEG peptides idlula kwi-glomerular capillary cross section ngokulula ngakumbi kuneepeptides eziqhelekileyo, kunciphisa kakhulu ukucocwa kwezintso.Ngenxa yolwandiso lwesiqingatha sobomi be-PEG peptides kwi-vivo, inqanaba lonyango eliqhelekileyo linokugcinwa ngeedosi ezisezantsi kunye neziyobisi zepeptide ezingaphantsi rhoqo.Nangona kunjalo, ukuguqulwa kwe-PEG kunemiphumo emibi.Izixa ezikhulu ze-PEG zithintela i-enzyme ekuthobeni i-peptide kwaye ikwanciphisa ukubophelela kwe-peptide kwi-receptor ekujoliswe kuyo.Kodwa ubudlelwane obuphantsi be-PEG peptides buhlala bulungiswa bubomi babo obude be-pharmacokinetic, kwaye ngokubakho emzimbeni ixesha elide, i-PEG peptides inethuba elikhulu lokufunxwa kwizihlunu ekujoliswe kuzo.Ke ngoko, iinkcukacha ze-PEG zepolymer kufuneka ziphuculwe ukuze zibe neziphumo ezizezona zilungileyo.Ngakolunye uhlangothi, i-PEG peptides iqokelela esibindini ngenxa yokunciphisa ukukhutshwa kwezintso, okukhokelela kwi-macromolecular syndrome.Ke ngoko, ukuguqulwa kwe-PEG kufuneka kuyilwe ngononophelo ngakumbi xa iipeptide zisetyenziselwa ukuvavanywa kweziyobisi.
Amaqela aqhelekileyo okuguqulwa kwe-PEG anokushwankathelwa ngolu hlobo lulandelayo: Amino (-amine) -NH2, aminomethyl-Ch2-NH2, hydroxy-OH, carboxy-Cooh, sulfhydryl (-Thiol) -SH, Maleimide -MAL, succinimide carbonate - SC, succinimide acetate -SCM, succinimide propionate -SPA, n-hydroxysuccinimide -NHS, Acrylate-ch2ch2cooh, aldehyde -CHO (efana nepropional-ald, butyrALD), isiseko se-acrylic (-acrylate-acrl), azido-azide, biotinyl - I-Biotin, i-Fluorescein, i-glutaryl -GA, i-Acrylate Hydrazide, i-alkyne-alkyne, i-p-toluenesulfonate -OTs, i-succinimide succinate -SS, njl. I-PEG derivatives kunye ne-carboxylic acids inokudibaniswa ne-n-terminal amines okanye i-lysine side chain.I-Amino-activated PEG inokudityaniswa ne-aspartic acid okanye i-glutamic side chain chain.I-PEG engasebenzi kakuhle inokudityaniswa kwi-mercaptan yeetyathanga zecala le-cysteine ezikhuselwe ngokupheleleyo [11].I-PEG izilungisi zidla ngokuhlelwa ngolu hlobo lulandelayo (qaphela: mPEG yi-methoxy-PEG, CH3O-(CH2CH2O)n-CH2CH2-OH) :
(1) ikhonkco elithe ngqo le-PEG isilungisi
mPEG-SC, mPEG-SCM, mPEG-SPA, mPEG-OTs, mPEG-SH, mPEG-ALD, mPEG-butyrALD, mPEG-SS
(2) isilungisi se-PEG esisebenza kabini
HCOO-PEG-COOH, NH2-PEG-NH2, OH-PEG-COOH, OH-PEG-NH2, HCl·NH2-PEG-COOH, MAL-PEG-NHS
(3) i-branching PEG isilungisi
(mPEG)2-NHS, (mPEG)2-ALD, (mPEG)2-NH2, (mPEG)2-MAL
8. Biotinization
I-Biotin inokubotshwa ngamandla kunye ne-avidin okanye i-streptavidin, kwaye amandla okubopha asondele ngakumbi kwi-covalent bond.Iipeptides ezibhalwe nge-Biotin ziqhele ukusetyenziswa kwi-immunoassay, i-histocytochemistry, kunye ne-fluorescence-based flow cytometry.Izilwa-buhlungu ezibhalwe i-antibiotin zingasetyenziselwa ukubopha iipeptide ze-biotinylated.Iileyibhile zeBiotin zihlala zincanyathiselwe kwikhonkco elisecaleni le-lysine okanye kwi-terminal ye-N.I-6-aminocaproic acid idla ngokusetyenziswa njengebhondi phakathi kweepeptides kunye ne-biotin.Ibhondi iguquguqukayo ekubopheleleni kwi-substrate kwaye ibophe ngcono xa kukho isithintelo se-steric.
9. Ileyibheli yeFluorescent
Ilebhile yeFluorescent ingasetyenziselwa ukulandelela iipolypeptides kwiiseli eziphilayo kunye nokufunda ii-enzymes kunye neendlela zokusebenza.I-Tryptophan (Trp) yi-fluorescent, ngoko ke ingasetyenziselwa ukuleyibhela ngaphakathi.I-spectrum yokukhutshwa kwe-tryptophan ixhomekeke kwimeko-bume ye-peripheral kwaye iyancipha ngokuncipha kwe-solvent polarity, ipropati eluncedo ekubhaqweni kwesakhiwo se-peptide kunye nokubopha i-receptor.I-Tryptophan fluorescence inokucinywa nge-protonated aspartic acid kunye ne-glutamic acid, enokunciphisa ukusetyenziswa kwayo.Iqela le-Dansyl chloride (i-Dansyl) i-fluorescent ephezulu xa ibotshelelwe kwiqela le-amino kwaye isoloko isetyenziswa njengeleyibhile ye-fluorescent yee-amino acids okanye iiprotheni.
I-Fluorescence resonance Ukuguqulwa kwamandla (FRET) luncedo kwizifundo ze-enzyme.Xa i-FRET isetyenziswa, i-substrate polypeptide idla ngokuba neqela le-fluorescence-labeling kunye neqela le-fluorescence-quenching.Amaqela e-fluorescent abhalwe ilebhile acinywa yi-quencher ngokugqithiswa kwamandla angeyi-photon.Xa i-peptide yahluliwe kwi-enzyme ekuthethwa ngayo, iqela lokulebula likhupha i-fluorescence.
10. Iipolypeptides zekheji
Iipeptide zeCage zinamaqela okhuselo asuswayo akhusela i-peptide ekubopheni i-receptor.Xa ibonakaliswe kwimitha ye-UV, i-peptide iyasebenza, ibuyisela ubudlelwane bayo kwi-receptor.Ngenxa yokuba oku kusebenza kwamehlo kunokulawulwa ngokwexesha, i-amplitude, okanye indawo, i-cage peptides inokusetyenziselwa ukufunda ukuphendula okwenzeka kwiiseli.Awona maqela axhaphakileyo asetyenziswayo akhuselayo kwi-cage polypeptides ngamaqela angama-2-nitrobenzyl kunye nezinto eziphuma kuzo, ezinokuthi zingeniswe kwi-peptide synthesis ngokukhuselwa kwe-amino acid derivatives.Iimveliso ze-amino acid eziye zaphuhliswa yi-lysine, i-cysteine, i-serine, kunye ne-tyrosine.I-aspartate kunye ne-glutamate derivatives, nangona kunjalo, azisetyenziswanga ngokuxhaphakileyo ngenxa yokuba semngciphekweni wokuhamba ngebhayisikile ngexesha le-peptide synthesis kunye ne-dissociation.
11. Polyantigenic peptide (MAP)
Iipeptide ezimfutshane zihlala zingakhuselekanga kwaye kufuneka zidityaniswe neeproteni ezithwala ukuvelisa izilwa-buhlungu.I-Polyantigenic peptide (MAP) yenziwe ngeepeptides ezininzi ezifanayo eziqhagamshelwe kwi-lysine nuclei, enokuthi iveze ngokuthe ngqo i-immunogens yamandla aphezulu kwaye ingasetyenziselwa ukulungisa i-peptide-carrier protein couplets.Iipolypeptides zeMAP zinokudityaniswa ngokudityaniswa kwesigaba esiqinileyo kwi-resin ye-MAP.Nangona kunjalo, ukudityaniswa okungaphelelanga kubangela amatyathanga epeptide alahlekileyo okanye acuthiweyo kwamanye amasebe kwaye ke ayibonisi iimpawu zepolypeptide yoqobo yeMAP.Njengenye indlela, iipeptides zinokulungiswa kwaye zicocwe ngokwahlukeneyo emva koko zidityaniswe kwi-MAP.Ulandelelwano lwepeptide oluncanyathiselwe kumbindi wepeptide lucaciswe kakuhle kwaye luphawulwa ngokulula nge-mass spectrometry.
Ukuqukumbela
Ukuguqulwa kweepeptide yindlela ebalulekileyo yokuyila iipeptides.Iipeptide eziguqulwe ngamachiza azikwazi ukugcina kuphela umsebenzi ophezulu webhayoloji, kodwa ziphinde ziphephe ngokufanelekileyo imiqobo ye-immunogenicity kunye netyhefu.Kwangaxeshanye, ukuguqulwa kweekhemikhali kunokunika iipeptides ngeempawu ezintsha ezibalaseleyo.Kwiminyaka yakutshanje, indlela ye-CH isebenze kwi-post-modification ye-polypeptides iphuhliswe ngokukhawuleza, kwaye iziphumo ezininzi ezibalulekileyo zifunyenwe.
Ixesha lokuposa: Mar-20-2023