Meiyuan Qian, Yuefeng Tan, Bo Xu
Isicelo se-R&D Center
Isingeniso
I-Taxus (i-Taxus chinensis noma i-Chinese yew) isitshalo sasendle esivikelwe yizwe.Kuyisitshalo esiyivelakancane nesisengozini esishiywe yizinguzunga zeqhwa ze-Quaternary.Yisona sodwa isitshalo sokwelapha esingokwemvelo emhlabeni.I-Taxus isatshalaliswa endaweni epholile yenyakatho nenkabazwe ukuya endaweni emaphakathi ne-subtropical, cishe ngezinhlobo eziyi-11 emhlabeni.Kunezinhlobo ezi-4 nezinhlobonhlobo ezi-1 e-China, okuyi-Northeast Taxus, i-Yunnan Taxus, i-Taxus, i-Tibetan Taxus kanye ne-Southern Taxus.Lezi zinhlobo ezinhlanu zisatshalaliswa eNingizimu-ntshonalanga yeChina, eNingizimu yeChina, eChina Emaphakathi, eMpumalanga China, eNyakatho-ntshonalanga yeChina, eNyakatho-mpumalanga yeChina naseTaiwan.Izitshalo ze-Taxus ziqukethe izinhlobonhlobo zamakhemikhali ahlukahlukene, okuhlanganisa ama-taxane, ama-flavonoid, ama-lignans, ama-steroid, ama-phenolic acid, ama-sesquiterpenes nama-glycosides.Isidakamizwa esidumile sokulwa nesimila i-Taxol (noma i-Paclitaxel) siwuhlobo lwe-taxane.I-Taxol inezindlela ezihlukile zokulwa nomdlavuza.I-Taxol "ingaqandisa" ama-microtubules ngokuhlangana nawo futhi ivimbele ama-microtubules ekuhlukaniseni ama-chromosome ngesikhathi sokuhlukana kwamaseli, ngaleyo ndlela kuholele ekufeni kwamangqamuzana ahlukanisayo, ikakhulukazi amangqamuzana omdlavuza akhula ngokushesha[1].Ngaphezu kwalokho, ngokwenza kusebenze ama-macrophage, i-Taxol ibangela ukwehla kwama-receptors e-TNF-α (i-tumor necrosis factor) kanye nokukhululwa kwe-TNF-α, ngaleyo ndlela ibulale noma ivimbele amaseli wesimila[2].Ngaphezu kwalokho, i-Taxol ingenza i-apoptosis ngokusebenza endleleni ye-apoptotic receptor mediat by Fas/FasL noma isebenze uhlelo lwe-cysteine protease[3].Ngenxa yomphumela wayo wokulwa nomdlavuza, i-Taxol isetshenziswa kabanzi ekwelapheni umdlavuza wesibeletho, umdlavuza webele, umdlavuza wamaphaphu ongewona omncane (NSCLC), umdlavuza wesisu, umdlavuza womminzo, umdlavuza wesinye, umdlavuza wesinye, umdlavuza wendlala yesinye, i-melanoma eyingozi, ikhanda nentamo. umdlavuza, njll[4].Ikakhulukazi umdlavuza webele othuthukile kanye nomdlavuza we-ovarian othuthukile, i-Taxol inomphumela omuhle kakhulu wokwelapha, ngakho-ke yaziwa ngokuthi "umugqa wokugcina wokuzivikela ekwelashweni komdlavuza".
I-Taxol isidakamizwa esilwa nomdlavuza esidume kakhulu emakethe yamazwe ngamazwe eminyakeni yamuva nje futhi ibhekwa njengenye yezidakamizwa ezisebenza kahle kakhulu zokulwa nomdlavuza kubantu eminyakeni engu-20 ezayo.Eminyakeni yamuva nje, ngokukhula okukhulu kwesibalo sabantu kanye nezigameko zomdlavuza, isidingo seTaxol sikhule kakhulu.Okwamanje, i-Taxol edingekayo ocwaningweni lwezokwelapha noma lwesayensi ikhishwa ikakhulukazi ku-Taxus.Ngeshwa, okuqukethwe kwe-Taxol ezitshalweni kuphansi kakhulu.Isibonelo, okuqukethwe kwe-Taxol kungu-0.069% kuphela emagxolweni e-Taxus brevifolia, ngokuvamile abhekwa njengokuqukethwe okuphezulu kakhulu.Ukuze kukhishwe i-1 g ye-Taxol, idinga cishe u-13.6 kg wegxolo le-Taxus.Ngokusekelwe kulokhu kulinganisa, kuthatha izihlahla ze-Taxus ezi-3 – 12 ezineminyaka engaphezu kweyi-100 ukwelapha isiguli somdlavuza wesibeletho.Ngenxa yalokho, inani elikhulu lezihlahla ze-Taxus ligawuliwe, okubangele ukushabalala kwalolu hlobo oluyigugu.Ngaphezu kwalokho, i-Taxus impofu kakhulu ezinsizeni futhi ihamba kancane ekukhuleni, okwenza kube nzima ukuqhubeka nokuthuthuka nokusetshenziswa kwe-Taxol.
Njengamanje, ukuhlanganiswa okuphelele kwe-Taxol kuqedwe ngempumelelo.Kodwa-ke, umzila wayo wokwenziwa uyinkimbinkimbi kakhulu futhi ubiza kakhulu, okwenza ungabi nokubaluleka kwezimboni.Indlela ye-semi-synthetic ye-Taxol manje isivuthiwe futhi ibhekwa njengendlela ephumelelayo yokwandisa umthombo we-Taxol ngaphezu kokutshala okuzenzela.Kafushane, ku-semi-synthesis ye-Taxol, isandulela se-Taxol esitholakala ngobuningi ezitshalweni ze-Taxus siyakhishwa bese siguqulelwa ku-Taxol ngokuhlanganiswa kwamakhemikhali.Okuqukethwe kwe-10-deacetylbaccatin Ⅲ ezinalitini ze-Taxus baccata kungaba kufika ku-0.1%.Futhi izinaliti kulula ukuvuselela ngokuqhathanisa namagxolo.Ngakho-ke, i-semi-synthesis ye-Taxol esekelwe ku-10-deacetylbaccatin Ⅲ idonsa ukunakwa okwengeziwe okuvela kubacwaningi[5] (njengoba kuboniswe kuMfanekiso 1).
Umfanekiso 1. Umzila we-semi-synthetic we-Taxol osuselwe ku-10-deacetylbaccatin Ⅲ.
Kulokhu okuthunyelwe, ukukhishwa kwesitshalo se-Taxus kuhlanzwe ngomshini we-chromatography owuketshezi olungiselelwe i-SepaBean™ ohlanganiswe nama-flash cartridges we-SepaFlash C18 reversed-phase (RP) akhiqizwe yi-Santai Technologies.Umkhiqizo ohlosiwe ohlangabezana nezidingo zobumsulwa watholwa futhi ungasetshenziswa ocwaningweni lwesayensi olulandelayo, unikeze isisombululo esingabizi kakhulu sokuhlanzwa okusheshayo kwalolu hlobo lwemikhiqizo yemvelo.
| Insimbi | SepaBean™ umshini | |
| Ikhatriji | 12 g SepaFlash C18 RP flash cartridge (silica eyindilinga, 20 - 45μm, 100 Å, inombolo ye-oda:SW-5222-012-SP) | |
| Ubude begagasi | 254 nm (ukutholwa), 280 nm (ukuqapha) | |
| Isigaba seselula | I-solvent A: Amanzi | |
| I-solvent B: I-Methanol | ||
| Izinga lokugeleza | 15 mL/min | |
| Isampula iyalayisha | 20 mg isampula eluhlaza incibilike ku-1 mL DMSO | |
| I-Gradient | Isikhathi (imizuzu) | I-solvent B (%) |
| 0 | 10 | |
| 5 | 10 | |
| 7 | 28 | |
| 14 | 28 | |
| 16 | 40 | |
| 20 | 60 | |
| 27 | 60 | |
| 30 | 72 | |
| 40 | 72 | |
| 43 | 100 | |
| 45 | 100 | |
Imiphumela nengxoxo
I-flash chromatogram yokukhishwa okungahluziwe okuvela ku-Taxus iboniswe kuMfanekiso 2. Ngokuhlaziya i-chromatogram, umkhiqizo okuhlosiwe kanye nokungcola kuzuze ukuhlukaniswa okuyisisekelo.Ngaphezu kwalokho, ukukhiqiza kabusha okuhle kuphinde kwabonakala ngemijovo yamasampula amaningi (idatha ayiboniswanga).Kuzothatha cishe amahora angu-4 ukuqedela ukuhlukanisa ngendlela ye-chromatography eyenziwa ngamakholomu engilazi.Uma kuqhathaniswa nendlela yendabuko ye-chromatography yamanuwali, indlela yokuhlanza okuzenzakalelayo kulokhu okuthunyelwe idinga kuphela imizuzu engama-44 ukuqedela wonke umsebenzi wokuhlanza (njengoba kuboniswe kuMfanekiso 3).Isikhathi esingaphezu kuka-80% kanye nenani elikhulu le-solvent lingasindiswa ngokuthatha indlela ezenzakalelayo, enganciphisa ngempumelelo izindleko kanye nokuthuthukisa kakhulu ukusebenza kahle komsebenzi.
Umfanekiso 2. I-flash chromatogram yokukhishwa okungahluziwe okuvela ku-Taxus.
Umfanekiso 3. Ukuqhathaniswa kwendlela ye-chromatography ye-manual kanye nendlela yokuhlanza okuzenzakalelayo.
Sengiphetha, ukuhlanganisa ama-flash cartridges e-SepaFlash C18 RP ngomshini we-SepaBean™ kunganikeza isisombululo esisheshayo nesisebenzayo sokuhlanzwa okusheshayo kwemikhiqizo yemvelo efana ne-Taxus extract.
Izithenjwa
1. I-Alushin GM, i-Lander GC, i-Kellogg EH, i-Zhang R, i-Baker D ne-Nogales E. Izakhiwo ze-microtubule zokucaca okuphezulu zibonisa ukuguqulwa kwesakhiwo ku-αβ-tubulin phezu kwe-GTP hydrolysis.Iseli, 2014, 157 (5), 1117-1129.
2. I-Burkhart CA, i-Berman JW, i-Swindell CS ne-Horwitz SB.Ubudlelwano phakathi kweSakhiwo Se-Taxol kanye Nezinye Izintela Ekungenweni Kwe-Tumor Necrosis Factor-α I-Gene Expression kanye ne-Cytotoxicity.Ucwaningo lweCancer, 1994, 54 (22), 5779-5782.
3. Ipaki SJ, Wu CH, Gordon JD, Zhong X, Emami A kanye neSafa AR.I-Taxol Idala i-Caspase-10-dependent Apoptosis, J. Biol.Chem., 2004, 279, 51057-51067.
4. I-Paclitaxel.I-American Society of Health-System Pharmacists.[Januwari 2, 2015]
5. UBruce Ganem noRoland R. Franke.I-Paclitaxel evela ku-Primary Taxanes: Umbono Wokuqanjwa Kwezinto Eziqanjiwe ku-Organozirconium Chemistry.J. Org.Chem., 2007, 72 (11), 3981-3987.
Kukhona uchungechunge lwama-flash cartridges e-SepaFlash C18 RP anemininingwane ehlukene evela ku-Santa Technology (njengoba kukhonjisiwe kuThebula 2).
| Inombolo Yento | Usayizi Wekholomu | Isilinganiso sokugeleza (mL/min) | Ubukhulu.Ingcindezi (i-psi/ibha) |
| I-SW-5222-004-SP | 5.4 g | 5-15 | 400/27.5 |
| I-SW-5222-012-SP | 20 g | 10-25 | 400/27.5 |
| I-SW-5222-025-SP | awu 33g | 10-25 | 400/27.5 |
| I-SW-5222-040-SP | awu 48g | 15-30 | 400/27.5 |
| I-SW-5222-080-SP | 105 g | 25-50 | 350/24.0 |
| I-SW-5222-120-SP | awu 155g | 30-60 | 300/20.7 |
| I-SW-5222-220-SP | 300 g | 40-80 | 300/20.7 |
| I-SW-5222-330-SP | awu 420g | 40-80 | 250/17.2 |
Ithebula 2. I-SepaFlash C18 RP flash cartridges.
Izinto zokupakisha: Isilika eyindilinga eyi-C18 esebenza kahle kakhulu, 20 - 45 μm, 100 Å
Ukuze uthole ulwazi olwengeziwe mayelana nemininingwane ecacisiwe yomshini we-SepaBean™, noma imininingwane yoku-oda kuma-flash cartridges ochungechunge lwe-SepaFlash, sicela uvakashele iwebhusayithi yethu.
Isikhathi sokuthumela: Sep-20-2018