Meiyuan Qian, Yuefeng Tan, Bo Xu
Setsi sa R&D sa Kopo
Selelekela
Taxus (Taxus chinensis kapa Chinese yew) ke semela se hlaha se sirelelitsoeng ke naha.Ke semela se sa tloaelehang le se kotsing ea ho fela se siiloeng ke Quaternary glaciers.Hape ke semela se le seng sa meriana ea tlhaho lefatšeng.Taxus e ajoa sebakeng se futhumetseng sa hemisphere e ka leboea ho ea bohareng ba tropike, ka mefuta e ka bang 11 lefatšeng.Ho na le mefuta e 4 le mofuta o le mong Chaena, e leng Northeast Taxus, Yunnan Taxus, Taxus, Tibetan Taxus le Southern Taxus.Mefuta ena e mehlano e ajoa ka Boroa-bophirima ho Chaena, Chaena Boroa, Chaena Bohareng, Chaena Bochabela, Chaena Leboea-bophirimela, Chaena Leboea-bochabela le Taiwan.Limela tsa Taxus li na le mefuta e mengata ea metsoako ea lik'hemik'hale, ho kenyelletsa le taxanes, flavonoids, lignans, steroids, phenolic acids, sesquiterpenes le glycosides.Sethethefatsi se tsebahalang sa anti-tumor Taxol (kapa Paclitaxel) ke mofuta oa li-taxtanes.Taxol e na le mekhoa e ikhethang ea anticancer.Taxol e ka "hatsetsa" li-microtubules ka ho kopana le tsona le ho thibela li-microtubules ho arola li-chromosome nakong ea karohano ea lisele, kahoo e lebisa lefung la lisele tse arohaneng, haholo-holo lisele tsa mofetše tse atang ka potlako[1].Ho feta moo, ka ho kenya li-macrophages, Taxol e baka ho fokotseha ha li-receptor tsa TNF-α (tumor necrosis factor) le ho lokolloa ha TNF-α, kahoo e bolaea kapa e thibela lisele tsa tumor[2].Ho feta moo, Taxol e ka baka apoptosis ka ho sebetsa tseleng ea apoptotic receptor e kenelletseng ke Fas/FasL kapa ho kenya tšebetsong cysteine protease system[3].Ka lebaka la phello ea eona e matla ea anticancer, Taxol e sebelisoa haholo kalafong ea mofetše oa mahe a bomme, mofetše oa matsoele, mofets'e o seng o monyane oa lisele tsa matšoafo (NSCLC), mofets'e oa mpa, mofets'e, mofets'e oa senya, mofets'e oa senya, melanoma e mpe, hlooho le molala. kankere, joalo-joalo[4].Haholo-holo mofetše oa matsoele o tsoetseng pele le mofets'e o tsoetseng pele oa mahe a mae, Taxol e na le phello e ntle ea ho phekola, ka hona e tsejoa e le "mohala oa ho qetela oa ts'ireletso bakeng sa kalafo ea mofets'e".
Taxol ke setlhare se thibelang mofetše ka ho fetesisa 'marakeng oa machaba lilemong tsa morao tjena mme se nkuoa e le se seng sa litlhare tse thibelang mofetše ho batho lilemong tse 20 tse tlang.Lilemong tsa morao tjena, ka kholo e phatlohileng ea palo ea batho le ts'oaetso ea mofets'e, tlhoko ea Taxol le eona e eketsehile haholo.Hajoale, Taxol e hlokehang bakeng sa lipatlisiso tsa bongaka kapa tsa mahlale e ntšoa haholo ho tsoa ho Taxus.Ka bomalimabe, litaba tsa Taxol limeleng li tlase haholo.Mohlala, litaba tsa Taxol ke 0.069% feela lekhapetleng la Taxus brevifolia, leo hangata le nkoang le na le litaba tse holimo.Bakeng sa ho ntša 1 g ea Taxol, e hloka hoo e ka bang 13.6 kg ea makhapetla a Taxus.Ho ipapisitsoe le khakanyo ena, ho nka lifate tse 3 - 12 tsa Taxus tse fetang lilemo tse 100 ho alafa mokuli oa mofetše oa mahe a bomme.Ka lebaka leo, palo e kholo ea lifate tsa Taxus e remiloe, e leng se bakileng hore e be haufi le ho timela ha mofuta ona oa bohlokoa.Ho feta moo, Taxus e futsanehile haholo ka lisebelisoa ebile e lieha ho hola, e leng se etsang hore ho be thata bakeng sa nts'etsopele le ts'ebeliso ea Taxol.
Hajoale, kakaretso ea pokello ea Taxol e phethiloe ka katleho.Leha ho le joalo, tsela ea eona ea maiketsetso e rarahane haholo ebile e theko e phahameng, e etsa hore e se be le bohlokoa ba indasteri.Mokhoa oa semi-synthetic oa Taxol hona joale o se o hōlile 'me o nkoa e le mokhoa o sebetsang oa ho atolosa mohloli oa Taxol ho phaella ho lemang ha maiketsetso.Ka bokhutšoanyane, ho semi-synthesis ea Taxol, motsoako oa pele oa Taxol o batlang o le ngata limela tsa Taxus oa ntšoa ebe o fetoloa Taxol ka motsoako oa lik'hemik'hale.Likahare tsa 10-deacetylbaccatin Ⅲ ka linaleng tsa Taxus baccata li ka fihla ho 0.1%.'Me li-needle li bonolo ho tsosolosa ha li bapisoa le makhapetla.Ka hona, semi-synthesis ea Taxol e thehiloeng ho 10-deacetylbaccatin Ⅲ e hohela tlhokomelo e eketsehileng ho tsoa ho bafuputsi[5] (joalo ka ha ho bonts'itsoe ho Setšoantšo sa 1).
Setšoantšo sa 1. Tsela ea semi-synthetic ea Taxol e thehiloeng ho 10-deacetylbaccatin Ⅲ.
Ka poso ena, sehlahisoa sa semela sa Taxus se hloekisitsoe ka mochini oa flash preparative liquid chromatography SepaBean™ o kopantsoeng le li-cartridge tsa SepaFlash C18 reversed-phase (RP) tse hlahisoang ke Santai Technologies.Sehlahisoa se lebisitsoeng ho finyella litlhoko tsa bohloeki se ile sa fumanoa 'me se ka sebelisoa lipatlisisong tse latelang tsa saense, ho fana ka tharollo e theko e tlaase bakeng sa tlhoekiso e potlakileng ea mofuta ona oa lihlahisoa tsa tlhaho.
| Seletsa | SepaBean™ mochini | |
| Cartridge | 12 g SepaFlash C18 RP flash cartridge (silika e chitja, 20 - 45μm, 100 Å, Nomoro ea taelo: SW-5222-012-SP) | |
| Wavelength | 254 nm (ho lemoha), 280 nm (ho beha leihlo) | |
| Mokhahlelo oa selefouno | Solvent A: Metsi | |
| Solvent B: Methanol | ||
| Sekhahla sa phallo | 15 mL/min | |
| Ho kenya mohlala | 20 mg sampole e tala e qhibilihisitsoeng ka 1 mL DMSO | |
| Gradient | Nako (mets) | Solvent B (%) |
| 0 | 10 | |
| 5 | 10 | |
| 7 | 28 | |
| 14 | 28 | |
| 16 | 40 | |
| 20 | 60 | |
| 27 | 60 | |
| 30 | 72 | |
| 40 | 72 | |
| 43 | 100 | |
| 45 | 100 | |
Liphetho le puisano
Flash chromatogram bakeng sa mokoloto o sa jeseng litheohelang ho tsoa ho Taxus e bontšitsoe ho Setšoantšo sa 2. Ka ho sekaseka chromatogram, sehlahisoa se lebeletsoeng le litšila li fihletse karohano ea mantlha.Ho feta moo, ho ikatisa hantle ho ile ha boela ha bonoa ka liente tse ngata tsa sampole (data e sa bonts'itsoeng).Ho tla nka lihora tse ka bang 4 ho phethela karohano ka mokhoa oa chromatography ka likholomo tsa khalase.Ha ho bapisoa le mokhoa oa setso oa chromatography, mokhoa oa ho itlhoekisa ka mokhoa o itekanetseng posong ena o hloka feela metsotso ea 44 ho qeta mosebetsi oohle oa tlhoekiso (joalokaha ho bontšitsoe setšoantšong sa 3).Ho feta 80% ea nako le palo e kholo ea solvent e ka bolokeha ka ho nka mokhoa o itekanetseng, o ka fokotsang litšenyehelo ka katleho hammoho le ho ntlafatsa haholo katleho ea mosebetsi.
Setšoantšo sa 2. Palesa ea chromatogram e tsoang ho Taxus.
Setšoantšo sa 3. Papiso ea mokhoa oa chromatography oa manual le mokhoa oa ho itlhoekisa ka mokhoa o itekanetseng.
Qetellong, ho kopanya li-cartridge tsa SepaFlash C18 RP tse nang le mochine oa SepaBean™ ho ka fana ka tharollo e potlakileng le e sebetsang bakeng sa tlhoekiso e potlakileng ea lihlahisoa tsa tlhaho tse kang Taxus extract.
Litšupiso
1. Alushin GM, Lander GC, Kellogg EH, Zhang R, Baker D le Nogales E. Mehaho ea microtubule e phahameng e senola liphetoho tsa sebopeho sa αβ-tubulin holim'a GTP hydrolysis.Cell, 2014, 157 (5), 1117-1129.
2. Burkhart CA, Berman JW, Swindell CS le Horwitz SB.Kamano pakeng tsa Sebopeho sa Taxol le Makhetho a Mang ka Ho Induction of Tumor Necrosis Factor-α Gene Expression le Cytotoxicity.Lipatlisiso tsa Kankere, 1994, 54 (22), 5779-5782.
3. Park SJ, Wu CH, Gordon JD, Zhong X, Emami A le Safa AR.Taxol e Etsa hore Caspase-10 e itšetlehileng ka Apoptosis, J. Biol.Chem., 2004, 279, 51057-51067.
4. Paclitaxel.Mokhatlo oa Amerika oa Bo-rakhoebo-Sisteme ea Bo-rakhoebo.[Pherekhong 2, 2015]
5. Bruce Ganem le Roland R. Franke.Paclitaxel ho tsoa ho Makhetho a Mathomo: Pono mabapi le Tlhahiso ea Boqapi ho Organozirconium Chemistry.J. Org.Chem., 2007, 72 (11), 3981-3987.
Ho na le letoto la li-cartridges tsa SepaFlash C18 RP tse nang le litlhaloso tse fapaneng ho tloha ho Santai Technology (joalokaha ho bontšitsoe ho Lethathamo la 2).
| Nomoro ea Ntho | Boholo ba Kholomo | Sekhahla sa Phallo (mL/mots) | Khatello (psi/bar) |
| SW-5222-004-SP | 5.4 g | 5-15 | 400/27.5 |
| SW-5222-012-SP | 20 g | 10-25 | 400/27.5 |
| SW-5222-025-SP | 33 g | 10-25 | 400/27.5 |
| SW-5222-040-SP | 48 g | 15-30 | 400/27.5 |
| SW-5222-080-SP | 105 g | 25-50 | 350/24.0 |
| SW-5222-120-SP | 155 g | 30-60 | 300/20.7 |
| SW-5222-220-SP | 300 g | 40-80 | 300/20.7 |
| SW-5222-330-SP | 420 g | 40-80 | 250/17.2 |
Letlapa la 2. SepaFlash C18 RP flash cartridges.
Lisebelisoa tsa ho paka: Silika e nang le ts'ebetso e phahameng ea spherical C18-bonded, 20 - 45 μm, 100 Å
Ho fumana lintlha tse ling mabapi le lintlha tse qaqileng tsa mochini oa SepaBean™, kapa tlhaiso-leseling ea ho odara ka li-cartridge tsa SepaFlash, ka kopo etela sebaka sa rona sa Marang-rang.
Nako ea poso: Sep-20-2018