CN102153605B - Method for purifying Mizoribine - Google Patents
Method for purifying Mizoribine Download PDFInfo
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- CN102153605B CN102153605B CN201110050467.3A CN201110050467A CN102153605B CN 102153605 B CN102153605 B CN 102153605B CN 201110050467 A CN201110050467 A CN 201110050467A CN 102153605 B CN102153605 B CN 102153605B
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- Prior art keywords
- mizoribine
- membrane
- resin
- molecular weight
- filtrate
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- HZQDCMWJEBCWBR-UUOKFMHZSA-N Mizoribine Chemical compound OC1=C(C(=O)N)N=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](CO)O1 HZQDCMWJEBCWBR-UUOKFMHZSA-N 0.000 title claims abstract description 75
- 229950000844 mizoribine Drugs 0.000 title claims abstract description 75
- 238000000034 method Methods 0.000 title claims abstract description 36
- 239000012528 membrane Substances 0.000 claims abstract description 57
- 239000011347 resin Substances 0.000 claims abstract description 26
- 229920005989 resin Polymers 0.000 claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 24
- 238000000855 fermentation Methods 0.000 claims abstract description 10
- 230000004151 fermentation Effects 0.000 claims abstract description 10
- 241000894006 Bacteria Species 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 30
- 239000000706 filtrate Substances 0.000 claims description 27
- 238000004587 chromatography analysis Methods 0.000 claims description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- 238000000746 purification Methods 0.000 claims description 16
- 238000001914 filtration Methods 0.000 claims description 15
- 238000001728 nano-filtration Methods 0.000 claims description 15
- 239000008213 purified water Substances 0.000 claims description 15
- 238000000108 ultra-filtration Methods 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- 239000003957 anion exchange resin Substances 0.000 claims description 9
- 239000012141 concentrate Substances 0.000 claims description 8
- 239000003929 acidic solution Substances 0.000 claims description 7
- 239000000919 ceramic Substances 0.000 claims description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 239000004952 Polyamide Substances 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000003637 basic solution Substances 0.000 claims description 3
- 229920001577 copolymer Polymers 0.000 claims description 3
- 229920002647 polyamide Polymers 0.000 claims description 3
- 229920006393 polyether sulfone Polymers 0.000 claims description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 3
- 239000002207 metabolite Substances 0.000 abstract 1
- 230000000813 microbial effect Effects 0.000 abstract 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- 239000012535 impurity Substances 0.000 description 7
- 230000008030 elimination Effects 0.000 description 5
- 238000003379 elimination reaction Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 238000011084 recovery Methods 0.000 description 4
- 238000002054 transplantation Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 239000013589 supplement Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 241000187747 Streptomyces Species 0.000 description 2
- 238000005377 adsorption chromatography Methods 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000004134 energy conservation Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 238000011027 product recovery Methods 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- AICIYIDUYNFPRY-UHFFFAOYSA-N 1,3-dihydro-2H-imidazol-2-one Chemical class O=C1NC=CN1 AICIYIDUYNFPRY-UHFFFAOYSA-N 0.000 description 1
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 241000186046 Actinomyces Species 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 239000006260 foam Substances 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 206010025135 lupus erythematosus Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000005554 pickling Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000009156 water cure Methods 0.000 description 1
Landscapes
- Saccharide Compounds (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a method for purifying Mizoribine, which comprises the following steps: pretreating a fermentation liquid generated by fermenting Mizoribine generating bacteria, enriching with resin, decoloring with a membrane, and concentrating to obtain the Mizoribine. Compared with the prior art, the method provided by the invention has the advantages of high product concentration, high product purity, high product activity, stable product quality, no secondary pollution and the like, can be used for large-scale popularization especially for microbial water-soluble metabolites, and has wide application prospects.
Description
[technical field]
The present invention relates to a kind of method of purification of biological field, relate in particular to a kind of method of purification of mizoribine.
[background technology]
Mizoribine (mizoribine; MZR) be a kind of imidazoles nucleosides being isolated for 1971 from the nutrient solution of mould; after in the Streptomyces of actinomyces (Actinomycetes) section belongs to the tunning of streptomyces brefeldianum, also have mizoribine; its chemistry 1-β-D ribofuranose-4-by name carbamyl-5 hydroxyl imidazoles inner salt is a kind of water-soluble cpds.In addition, mizoribine is synthetic inhibited to the purine in nucleic acid metabolism, it enters after body cell, can suppress competitively IMP desaturase and GMP synthetic enzyme, intracellular GMP is reduced, the synthetic minimizing of nucleic acid, thereby the propagation that further suppresses cell, therefore, mizoribine can be used as a kind of novel immunosuppressor, from in December, 1991, in Japan, be applied to clinical renal transplantation, using it, the routine immunization after renal transplantation suppresses medicine at the many clinical transplantations of Japan center, China is also used for clinical in recent years using it as the anti-repulsion medicine of renal transplantation, and mizoribine can be used for treating systemic lupus erythematosus, rheumatoid arthritis, the autoimmune disorder such as nephrotic syndrome and systemic lupus erythematous.
The production method of mizoribine mainly contains microbe fermentation method and chemical synthesis both at home and abroad at present.The chemosynthesis step of mizoribine is many, severe reaction conditions, and the cycle is long, when carrying out the condensation reaction of base and ribose groups, also needs to carry out the protection of group and goes protection, thereby cause the total recovery of building-up reactions on the low side simultaneously, and cost is high; And microorganism fermentative production mizoribine has the features such as production process green, cost be low, is the production ways of an economic environmental protection.But in fermenting process, produced a large amount of auburn pigments, and unstable to light and heat, therefore need to it, remove impurity, decolouring, concentration by appropriate means.Existing mizoribine product processes adopts silica gel discolor element and destainer vacuum concentration, but actual effect is undesirable: wherein silica gel has stronger adsorption to mizoribine; And vacuum concentration needs operation under certain temperature and causes product further to be degraded, and causes product yield greatly to decline, and therefore need to find more effective method of purification.
[summary of the invention]
Technical problem to be solved by this invention is to provide a kind of method of purification of mizoribine, has the advantages such as preparation method is simple to operate, purification efficiency is high, cycles of concentration is higher and have good stability.
The present invention solves the problems of the technologies described above by the following technical programs: a kind of method of purification of mizoribine, produces bacterium by mizoribine and adopt following steps to operate through the fermented liquid of fermentation generation:
(1) fermentation liquor pretreatment: get appropriate mizoribine fermented liquid, and with basic solution, its pH value is adjusted to 8.0~10.0, then adopt filtering membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate;
(2) resin concentration: first the speed with 0.01~0.1 times of packed bed volume of per minute pumps in chromatography column by the mizoribine filtrate obtaining in (1), and has loaded pretreated strongly basic anion exchange resin in this chromatography column; Then adopt purified water that the solution in chromatography column is washed till to neutrality, with acidic solution, carry out wash-out more afterwards, and collect pregnant solution, obtain mizoribine pregnant solution;
(3) film decolouring is concentrated: the mizoribine pregnant solution obtaining in will (2) is by the ultra-filtration membrane processing of decolouring, and during this process, adds the purified water of 1~5 times of pregnant solution volume, and collection membrane filtrate, contains the effluent liquid of mizoribine; Then this filtrate is adopted nanofiltration membrane to concentrate to obtain mizoribine;
Wherein, described filtering membrane is ceramic membrane, and molecular weight cut-off is 10KD~50KD; Described strongly basic anion exchange resin is that skeleton is with amino [N-(CH of season on styrene-divinylbenzene copolymer
3)
2c
2h
4oH] anionite-exchange resin, or D201 resin, or D241 resin, or D246 resin; Described ultra-filtration membrane is poly (ether-sulfone) ultrafiltration membrane, and its molecular weight cut-off is 1KD~5KD; Described nanofiltration membrane is polyamide nanofiltration membrane, and molecular weight cut-off is 100D~1KD.
Further, the preferred molecular weight cut-off that described filtering membrane is ceramic membrane is 30KD.
Further, described acidic solution is a kind of of hydrochloric acid or phosphoric acid or acetic acid.
Further, the preferred molecular weight cut-off of described ultra-filtration membrane is 1KD.
Further, the preferred molecular weight cut-off of described nanofiltration membrane is 300D.
The beneficial effect of the method for purification of a kind of mizoribine of the present invention is: preparation method is simple to operate, purification efficiency is high; In fermentation liquor pretreatment, decolouring concentration process, adopt membrane technique, adopted membrane filtration, film decolouring to concentrate, there is the advantages such as efficient, energy-conservation, process is easy to control, easy to operate, less contamination; Mizoribine fermented liquid (being Vandyke brown), through the adsorption chromatography of strongly basic anion exchange resin, makes target product be able to enrichment; Film decolouring impurity elimination obtains pale yellow solution after processing, and further membrane concentration, and the final cycles of concentration of mizoribine is higher, and product recovery rate reaches more than 85%, and in addition, this preparation method has satisfactory stability, and can carry out large-scale promotion application.
[embodiment]
The method of purification of a kind of mizoribine of the present invention, mizoribine is produced to bacterium adopts following steps to operate through the fermented liquid of fermentation generation: (1) fermentation liquor pretreatment: get appropriate mizoribine fermented liquid, and with basic solution, its pH value is adjusted to 8.0~10.0, then adopt filtering membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate; (2) resin concentration: first the speed with 0.01~0.1 times of packed bed volume of per minute pumps in chromatography column by the mizoribine filtrate obtaining in (1), and has loaded pretreated strongly basic anion exchange resin in this chromatography column; Then adopt purified water that the solution in chromatography column is washed till to neutrality, with acidic solution, carry out wash-out more afterwards, and collect pregnant solution, obtain mizoribine pregnant solution; (3) film decolouring is concentrated: the mizoribine pregnant solution obtaining in will (2) is by the ultra-filtration membrane processing of decolouring, and during this process, adds the purified water of 1~5 times of pregnant solution volume, and collection membrane filtrate, contains the effluent liquid of mizoribine; Then this filtrate is adopted nanofiltration membrane to concentrate to obtain mizoribine.In addition, in the present invention, it is with amino [N-(CH of season on styrene-divinylbenzene copolymer that strongly basic anion exchange resin can be selected skeleton
3)
2c
2h
4oH] anionite-exchange resin, D201 resin, D241 resin, D246 resin in any; Acidic solution is a kind of of hydrochloric acid or phosphoric acid or acetic acid; Filtering membrane adopts ceramic membrane, and its molecular weight cut-off is 10KD~50KD; Ultra-filtration membrane is poly (ether-sulfone) ultrafiltration membrane, and its molecular weight cut-off is 1KD~5KD; Nanofiltration membrane is polyamide nanofiltration membrane, and molecular weight cut-off is 100D~1KD.
For better the present invention will be described, the present invention is for following several embodiment, and in each embodiment, better for the effect that makes to process, the preferred molecular weight cut-off of filtering membrane is that 30KD, the preferred molecular weight cut-off of ultra-filtration membrane are that 1KD, the preferred molecular weight cut-off of nanofiltration membrane are 300D, because hydrochloric acid consumption is few, overall price is cheap, and acidic solution is selected hydrochloric acid, strongly basic anion exchange resin adopts more easily purchases available D201 resin, and it is to be produced by the Shanghai China scientific and technological trading company of shake, in addition, in order to reach to remove, to be filled in chromatography column the impurity in D201 resin and to activate the objects such as its ion, need carry out pre-treatment to resin, what adopt is conventional treatment process, concrete operations are: by preparing D201 resin that dress post uses, first use 50-60 ℃ of hot water (clean tap water also can) repeatedly to clean, embathe to embathing water and be not with brown, foam seldom time till, then the D201 resin of 5L hot-water cure being crossed packs in the chromatography column of Φ 10 * 120cm, afterwards resin is carried out to alkali cleaning (adopting 10 liter of 5% soaking with sodium hydroxide 24 hours), then by purified water, be washed till neutrality, then pickling (adopting 10 liters of 1.0mol/L salt acid soak 24 hours), by purified water, be washed till neutrality again, carry out again afterwards alkali cleaning (adopting 10 liter of 5% soaking with sodium hydroxide 24 hours), finally by purified water, be washed till neutrality, obtain pretreated D201 resin, standby.
Embodiment mono-
Get appropriate mizoribine fermented liquid and adjust pH to 8.0 with sodium hydroxide, and adopt ceramic membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate; Then getting 50L mizoribine filtrate pumps in the chromatography column of having loaded pretreated D201 resin with the speed of per minute 100ml, after filtrate all pumps into, adopt purified water drip washing chromatography column to neutral, with 0.05% hydrochloric acid, carry out wash-out more afterwards, and collect pregnant solution, finally obtain 20L mizoribine pregnant solution; Again 20L mizoribine pregnant solution being poured into the impurity elimination of decolouring in ultra-filtration membrane tank processes, and during this process, supplement purified water 60L, and collection membrane filtrate (is light yellow, contain the effluent liquid of mizoribine), adopt nanofiltration membrane to concentrate this filtrate, obtain concentrated solution 4L, mizoribine total recovery 91.2%.
Embodiment bis-
Get appropriate mizoribine fermented liquid and adjust pH to 10.0 with sodium hydroxide, and adopt ceramic membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate; Then getting 25L mizoribine filtrate pumps in the chromatography column of having loaded pretreated D201 resin with the speed of per minute 80ml, after filtrate all pumps into, adopt purified water drip washing chromatography column to neutral, with 0.05% hydrochloric acid, carry out wash-out more afterwards, and collect pregnant solution, finally obtain 15L mizoribine pregnant solution; Again 15L mizoribine pregnant solution being poured into the impurity elimination of decolouring in ultra-filtration membrane tank processes, and during this process, supplement purified water 40L, and collection membrane filtrate (is light yellow, contain the effluent liquid of mizoribine), adopt nanofiltration membrane to concentrate this filtrate, obtain concentrated solution 3.5L, mizoribine total recovery 88.6%.
Embodiment tri-
Get appropriate mizoribine fermented liquid and adjust pH to 9.0 with sodium hydroxide, and adopt ceramic membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate; Then getting 40L mizoribine filtrate pumps in the chromatography column of having loaded pretreated D201 resin with the speed of per minute 100ml, after filtrate all pumps into, adopt purified water drip washing chromatography column to neutral, with 0.05% hydrochloric acid, carry out wash-out more afterwards, and collect pregnant solution, finally obtain 20L mizoribine pregnant solution; Again 20L mizoribine pregnant solution being poured into the impurity elimination of decolouring in ultra-filtration membrane tank processes, and during this process, supplement purified water 50L, and collection membrane filtrate (is light yellow, contain the effluent liquid of mizoribine), adopt nanofiltration membrane to concentrate this filtrate, obtain concentrated solution 4L, mizoribine total recovery 90.6%.
To sum up, preparation method of the present invention is simple to operate, purification efficiency is high; In fermentation liquor pretreatment, decolouring concentration process, adopt membrane technique, adopted membrane filtration, film decolouring to concentrate, there is the advantages such as efficient, energy-conservation, process is easy to control, easy to operate, less contamination; Mizoribine fermented liquid (being Vandyke brown), through the adsorption chromatography of strongly basic anion exchange resin, makes target product enrichment; Film decolouring impurity elimination obtains pale yellow solution after processing, and further membrane concentration, and the final cycles of concentration of mizoribine is higher, can reach more than 10 times, and product recovery rate reaches more than 85%, in addition, this preparation method has satisfactory stability, and can carry out large-scale promotion application.
Claims (5)
1. a method of purification for mizoribine, is characterized in that: mizoribine is produced to the fermented liquid that bacterium produces through fermentation and adopt following steps to operate:
(1) fermentation liquor pretreatment: get appropriate mizoribine fermented liquid, and with basic solution, its pH value is adjusted to 8.0~10.0, then adopt filtering membrane to carry out conventional filtration this fermented liquid that mixes up pH value, thereby obtain mizoribine filtrate;
(2) resin concentration: first the speed with 0.01~0.1 times of packed bed volume of per minute pumps in chromatography column by the mizoribine filtrate obtaining in (1), and has loaded pretreated strongly basic anion exchange resin in this chromatography column; Then adopt purified water that the solution in chromatography column is washed till to neutrality, with acidic solution, carry out wash-out more afterwards, and collect pregnant solution, obtain mizoribine pregnant solution;
(3) film decolouring is concentrated: the mizoribine pregnant solution obtaining in will (2) is by the ultra-filtration membrane processing of decolouring, and during this process, adds the purified water of 1~5 times of pregnant solution volume, and collection membrane filtrate, contains the effluent liquid of mizoribine; Then this filtrate is adopted nanofiltration membrane to concentrate to obtain mizoribine;
Wherein, described filtering membrane is ceramic membrane, and its molecular weight cut-off is 10KD~50KD; Described strongly basic anion exchange resin is that skeleton is with amino [N-(CH of season on styrene-divinylbenzene copolymer
3)
2c
2h
4oH] anionite-exchange resin, or D201 resin, or D241 resin, or D246 resin; Described ultra-filtration membrane is poly (ether-sulfone) ultrafiltration membrane, and its molecular weight cut-off is 1KD~5KD; Described nanofiltration membrane is polyamide nanofiltration membrane, and molecular weight cut-off is 100D~1KD.
2. the method for purification of a kind of mizoribine as claimed in claim 1, is characterized in that: the preferred molecular weight cut-off of described filtering membrane is 30KD.
3. the method for purification of a kind of mizoribine as claimed in claim 1, is characterized in that: described acidic solution is a kind of of hydrochloric acid or phosphoric acid or acetic acid.
4. the method for purification of a kind of mizoribine as claimed in claim 1, is characterized in that: the preferred molecular weight cut-off of described ultra-filtration membrane is 1KD.
5. the method for purification of a kind of mizoribine as claimed in claim 1, is characterized in that: the preferred molecular weight cut-off of described nanofiltration membrane is 300D.
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CN201110050467.3A CN102153605B (en) | 2011-03-02 | 2011-03-02 | Method for purifying Mizoribine |
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CN201110050467.3A CN102153605B (en) | 2011-03-02 | 2011-03-02 | Method for purifying Mizoribine |
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CN102153605B true CN102153605B (en) | 2014-02-19 |
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CN103450300B (en) * | 2013-09-12 | 2015-10-28 | 华北制药集团新药研究开发有限责任公司 | A kind of method of purification of mizoribine |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1341124A (en) * | 1999-01-15 | 2002-03-20 | 诺瓦提斯公司 | Anti-CD3 immunotoxins and therapeutic use thereof |
CN1465410A (en) * | 2002-06-27 | 2004-01-07 | 微创医疗器械(上海)有限公司 | Drug-eluting stent (DES) with multicoating |
CN1478087A (en) * | 2000-11-07 | 2004-02-25 | ��˹��ŵ�� | Indolylmaleimide derivatives as protein kinase C inibitors |
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2011
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1341124A (en) * | 1999-01-15 | 2002-03-20 | 诺瓦提斯公司 | Anti-CD3 immunotoxins and therapeutic use thereof |
CN1478087A (en) * | 2000-11-07 | 2004-02-25 | ��˹��ŵ�� | Indolylmaleimide derivatives as protein kinase C inibitors |
CN1465410A (en) * | 2002-06-27 | 2004-01-07 | 微创医疗器械(上海)有限公司 | Drug-eluting stent (DES) with multicoating |
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