Meiyuan Qian, Yuefeng Tan, Bo Xu
Daim ntawv thov R&D Center
Taw qhia
Taxus (Taxus chinensis los yog Suav yew) yog cov nroj tsuag qus uas tiv thaiv lub teb chaws.Nws yog ib tsob nroj uas tsis tshua muaj thiab muaj kev puas tsuaj uas tau tso tseg los ntawm Quaternary glaciers.Nws kuj yog tib yam tshuaj ntsuab nroj tsuag hauv ntiaj teb no.Taxus tau faib rau thaj tsam huab cua ntawm sab qaum teb hemisphere mus rau thaj tsam nruab nrab-subtropical, nrog txog 11 hom hauv ntiaj teb.Muaj 4 hom thiab 1 ntau yam nyob rau hauv Suav teb, xws li Northeast Taxus, Yunnan Taxus, Taxus, Tibetan Taxus thiab Southern Taxus.Cov tsib hom no tau muab faib rau Southwest China, South China, Central Tuam Tshoj, East Tuam Tshoj, Northwest Tuam Tshoj, Northeast Tuam Tshoj thiab Taiwan.Taxus cov nroj tsuag muaj ntau yam tshuaj lom neeg, suav nrog setanes, flavonoids, lignans, steroids, phenolic acids, sesquiterpenes thiab glycosides.Cov tshuaj tiv thaiv kab mob nto moo Taxol (los yog Paclitaxel) yog ib hom sedanes.Taxol muaj cov tshuaj tiv thaiv kab mob tshwj xeeb.Taxol tuaj yeem "kho" microtubules los ntawm kev sib txuas nrog lawv thiab tiv thaiv cov microtubules los ntawm kev sib cais chromosomes thaum lub sijhawm sib faib ntawm tes, yog li ua rau kev tuag ntawm kev faib hlwb, tshwj xeeb tshaj yog cov qog nqaij hlav cancer sai sai[1].Tsis tas li ntawd, los ntawm kev ua kom cov macrophages, Taxol ua rau txo qis hauv TNF-α (cov qog necrosis factor) receptors thiab tso tawm ntawm TNF-α, yog li tua lossis inhibiting qog hlwb[2].Ntxiv mus, Taxol tuaj yeem ua rau apoptosis los ntawm kev ua ntawm apoptotic receptor txoj kev kho los ntawm Fas / FasL lossis ua kom lub cev cysteine protease [3].Vim nws ntau lub hom phiaj anticancer nyhuv, Taxol yog dav siv nyob rau hauv kev kho mob ntawm zes qe menyuam cancer, cancer ntawm lub mis, uas tsis yog-me me cell lung cancer (NSCLC), gastric cancer, esophageal cancer, zais zis cancer, prostate cancer, malignant melanoma, lub taub hau thiab caj dab. mob cancer, thiab lwm yam [4].Tshwj xeeb tshaj yog rau mob qog noj ntshav siab thiab mob qog noj ntshav ntawm zes qe menyuam, Taxol muaj cov txiaj ntsig zoo, yog li nws yog lub npe hu ua "kab kawg ntawm kev tiv thaiv kev kho mob qog noj ntshav".
Taxol yog cov tshuaj tiv thaiv kab mob uas nrov tshaj plaws hauv kev lag luam thoob ntiaj teb hauv xyoo tas los no thiab suav tias yog ib qho tshuaj tiv thaiv kab mob zoo tshaj plaws rau tib neeg hauv 20 xyoo tom ntej.Nyob rau hauv xyoo tas los no, nrog kev loj hlob ntawm cov pej xeem thiab kev mob qog noj ntshav, qhov kev thov rau Taxol kuj tau nce ntxiv.Tam sim no, Taxol xav tau rau kev kho mob lossis kev tshawb fawb tshawb fawb feem ntau yog muab rho tawm ncaj qha los ntawm Taxus.Hmoov tsis zoo, cov ntsiab lus ntawm Taxol hauv cov nroj tsuag yog tsawg heev.Piv txwv li, Taxol cov ntsiab lus tsuas yog 0.069% hauv cov tawv ntoo ntawm Taxus brevifolia, uas feem ntau suav hais tias muaj cov ntsiab lus siab tshaj plaws.Rau kev rho tawm ntawm 1 g ntawm Taxol, nws xav tau txog 13.6 kg ntawm Taxus bark.Raws li qhov kev kwv yees no, nws yuav siv 3 - 12 Taxus ntoo uas muaj hnub nyoog tshaj 100 xyoo los kho tus neeg mob qog noj ntshav zes qe menyuam.Raws li qhov tshwm sim, ntau tus ntoo Taxus tau raug txiav, uas ua rau ze rau qhov kawg ntawm cov tsiaj no.Tsis tas li ntawd, Taxus yog cov neeg pluag heev hauv cov peev txheej thiab kev loj hlob qeeb, uas ua rau nws nyuaj rau kev txhim kho ntxiv thiab kev siv Taxol.
Tam sim no, tag nrho cov synthesis ntawm Taxol tau ua tiav tiav.Txawm li cas los xij, nws txoj kev hluavtaws yog qhov nyuaj heev thiab tus nqi siab, ua rau nws tsis muaj qhov tseem ceeb hauv kev lag luam.Cov txheej txheem semi-synthetic ntawm Taxol yog tam sim no kuj paub tab thiab pom tau tias yog ib txoj hauv kev zoo los nthuav qhov chaw ntawm Taxol ntxiv rau kev cog qoob loo.Luv luv, nyob rau hauv lub semi-synthesis ntawm Taxol, lub Taxol precursor compound uas yog kuj muaj nyob rau hauv Taxus nroj tsuag yog muab rho tawm thiab ces hloov dua siab tshiab rau hauv Taxol los ntawm cov tshuaj synthesis.Cov ntsiab lus ntawm 10-deacetylbaccatin Ⅲ hauv koob ntawm Taxus baccata tuaj yeem nce mus txog 0.1%.Thiab cov koob yog ib qho yooj yim rau rov tsim dua piv nrog cov tawv ntoo.Yog li ntawd, lub semi-synthesis ntawm Taxol raws li 10-deacetylbaccatin Ⅲ yog attracting ntau thiab ntau xim los ntawm cov kws tshawb fawb[5] (raws li qhia nyob rau hauv daim duab 1).
Daim duab 1. Txoj kev semi-synthetic ntawm Taxol raws li 10-deacetylbaccatin Ⅲ.
Hauv cov ntawv tshaj tawm no, Taxus cog extract tau ntxuav los ntawm flash npaj ua kua chromatography system SepaBean ™ tshuab ua ke nrog SepaFlash C18 thim rov qab-theem (RP) flash cartridges tsim los ntawm Santai Technologies.Cov khoom lag luam lub hom phiaj ua tau raws li qhov xav tau purity tau txais thiab tuaj yeem siv rau hauv kev tshawb fawb tom ntej, muab cov txiaj ntsig zoo rau kev ua kom huv sai ntawm cov khoom ntuj no.
| Ntsuas | SepaBean ™ tshuab | |
| Cartridge | 12g SepaFlash C18 RP flash cartridge (spherical silica, 20 - 45μm, 100 Å, Order naj npawb: SW-5222-012-SP) | |
| Lub wavelength | 254 nm (nrhiav), 280 nm (saib xyuas) | |
| Mobile theem | Solvent A: Dej | |
| Cov kuab tshuaj B: Methanol | ||
| Tus nqi ntws | 15 ml / min | |
| Sample loading | 20 mg raw qauv yaj hauv 1 mL DMSO | |
| Gradient | Sijhawm (min) | Cov kuab tshuaj B (%) |
| 0 | 10 | |
| 5 | 10 | |
| 7 | 28 | |
| 14 | 28 | |
| 16 | 40 | |
| 20 | 60 | |
| 27 | 60 | |
| 30 | 72 | |
| 40 | 72 | |
| 43 | 100 | |
| 45 | 100 | |
Cov txiaj ntsig thiab kev sib tham
Lub flash chromatogram rau crude extract los ntawm Taxus tau pom nyob rau hauv daim duab 2. Los ntawm kev soj ntsuam cov chromatogram, lub hom phiaj cov khoom thiab cov impurities tiav hauv paus sib cais.Tsis tas li ntawd, kev tsim tawm zoo kuj tau pom los ntawm ntau cov qauv txhaj tshuaj (cov ntaub ntawv tsis qhia).Nws yuav siv sijhawm li 4 teev los ua kom tiav qhov kev sib cais hauv phau ntawv chromatography txoj kev nrog iav kab.Muab piv nrog cov txheej txheem ntawm phau ntawv chromatography ib txwm, txoj kev purification tsis siv neeg nyob rau hauv no ncej tsuas yog yuav tsum tau 44 feeb los ua kom tiav tag nrho cov hauj lwm purification (raws li qhia nyob rau hauv daim duab 3).Ntau tshaj 80% ntawm lub sijhawm thiab cov kuab tshuaj ntau tuaj yeem txuag tau los ntawm kev siv cov txheej txheem tsis siv neeg, uas tuaj yeem txo tus nqi kom zoo thiab txhim kho kev ua haujlwm zoo.
Daim duab 2. Lub flash chromatogram ntawm crude extract los ntawm Taxus.
Daim duab 3. Kev sib piv ntawm phau ntawv chromatography txoj kev nrog tsis siv neeg purification txoj kev.
Hauv kev xaus, combing SepaFlash C18 RP flash cartridges nrog SepaBean ™ tshuab tuaj yeem muab kev daws teeb meem sai thiab muaj txiaj ntsig rau kev ua kom huv sai ntawm cov khoom ntuj xws li Taxus extract.
Cov ntaub ntawv
1. Alushin GM, Lander GC, Kellogg EH, Zhang R, Baker D thiab Nogales E. High-resolution microtubule qauv qhia cov qauv hloov pauv hauv αβ-tubulin raws li GTP hydrolysis.Cell, 2014, 157 (5), 1117-1129.
2. Burkhart CA, Berman JW, Swindell CS thiab Horwitz SB.Kev sib raug zoo ntawm Cov Qauv ntawm Taxol thiab Lwm Cov Taxanes ntawm Induction ntawm Tumor Necrosis Factor-α Gene Expression thiab Cytotoxicity.Cancer Research, 1994, 54 (22), 5779-5782.
3. Park SJ, Wu CH, Gordon JD, Zhong X, Emami A, Safa AR.Taxol Induces Caspase-10-dependent Apoptosis, J. Biol.Chem., 2004, 279, 51057-51067.
4. Paclitaxel.American Society of Health-System Pharmacists.[Lub Ib Hlis 2, 2015]
5. Bruce Ganem thiab Roland R. Franke.Paclitaxel los ntawm Primary Taxanes: Kev Pom Zoo ntawm Kev Tsim Nyog Muaj Peev Xwm hauv Organozirconium Chemistry.J. Org.Chem., 2007, 72 (11), 3981-3987.
Muaj ib tug series ntawm SepaFlash C18 RP flash cartridges nrog txawv specifications ntawm Santai Technology (raws li qhia nyob rau hauv Table 2).
| Yam khoom naj npawb | Kem Loj | Tus nqi ntws (ml / min) | Max. Siab (psi/bar) |
| SW-5222-004-SP | 5,4g e | 5-15 | 400/27.5 |
| SW-5222-012-SP | 20 g ib | 10-25 | 400/27.5 |
| SW-5222-025-SP | 33g ua | 10-25 | 400/27.5 |
| SW-5222-040-SP | 48g ua | 15-30 | 400/27.5 |
| SW-5222-080-SP | 10 5g | 25-50 | 350/24.0 Nws |
| SW-5222-120-SP | 15 5g | 30-60 | 300/20.7 |
| SW-5222-220-SP | 300 g | 40-80 : kuv | 300/20.7 |
| SW-5222-330-SP | 420g ua | 40-80 : kuv | 250/17.2 |
Table 2. SepaFlash C18 RP flash cartridges.
Cov ntaub ntawv ntim: High-efficiency spherical C18-bonded silica, 20 - 45 μm, 100 Å
Yog xav paub ntxiv txog cov lus qhia ntxaws ntxaws ntawm SepaBean ™ tshuab, lossis cov ntaub ntawv xaj ntawm SepaFlash series flash cartridges, thov mus saib peb lub vev xaib
Post lub sij hawm: Sep-20-2018