CN107619473A - It is grafted the minimizing technology of DMAP in cholesterol amphipathy macromolecule materials synthesis - Google Patents
It is grafted the minimizing technology of DMAP in cholesterol amphipathy macromolecule materials synthesis Download PDFInfo
- Publication number
- CN107619473A CN107619473A CN201610550318.6A CN201610550318A CN107619473A CN 107619473 A CN107619473 A CN 107619473A CN 201610550318 A CN201610550318 A CN 201610550318A CN 107619473 A CN107619473 A CN 107619473A
- Authority
- CN
- China
- Prior art keywords
- cholesterol
- dmap
- grafting
- ester
- polyethylene glycol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention belongs to polymeric material field, and in particular to DMAP minimizing technology in a variety of grafting cholesterol amphipathy macromolecule materials synthesis.Removal DMAP of the present invention specific preparation method is as follows：（1）Take grafting cholesterol Amphiphatic high polymer material to be dissolved in toluene, be heated to 25 ~ 100oUltrasound makes its dissolving after C.（2）Rotating speed is used to centrifuge DMAP cholesterol intermediate for 1000 ~ 15000 rpm and be grafted the toluene solution of cholesterol Amphiphatic high polymer material.（3）Toluene solution layer is taken, sterling is produced after toluene is removed under reduced pressure.The advantage of the invention is that：（1）Removal available for DMAP in the amphipathy macromolecule material of a variety of grafting cholesterol；（2）The toluene precipitation method are simple to operate, purification efficiency is high, result is controllable, are advantageous to industrialized production.
Description
Technical field
The invention belongs to polymeric material field, and in particular in a variety of grafting cholesterol amphipathy macromolecule materials synthesis
DMAP minimizing technology.
Background technology
DMAP (4-Dimethylaminopyridine, DMAP) is efficiently urging in a kind of chemical synthesis
Agent, available for the speed and yield for improving a variety of acylation reactions, such as acyl chlorides, acid anhydrides, carboxylic acid and alcohol, phenol react into ester,
STEGLICHW, H FLE G.4-Dimethylamino-pyridin, with the amidation process of amine (ein hochwirksamer
Acylierungskatalysator.Angewandte Chemie,1969,81(23):1001-1001.)(SCRIVEN E
F.4-Dialkylaminopyridines:super acylation and alkylation catalysts.Chemical
Society Reviews,1983,12(2):129-161.).By taking acid anhydrides and alcohol reaction generation ester as an example, DMAP catalytic mechanism
For the carbon atom on 4 nitrogen-atoms nucleophilic attack acid anhydrides carbonyls on pyridine ring, DMAP- acid anhydrides intermediates are generated.Then, alcohol hydroxyl
Carbon atom on the oxygen atom nucleophilic attack DMAP- acid anhydrides intermediate carbonyls of base, ester bond generates (H FLE while DMAP leaves away
G,STEGLICH W,VORBRGGEN H.4‐Dialkylaminopyridines as Highly Active Acylation
Catalysts.[New synthetic method(25).Angewandte Chemie International Edition
in English,1978,17(8):569-583.)(GRONDAL C.4-Dimethylamino-pyridine(DMAP)
.Synlett,2003,2003(10):1568-1569.).Because DMAP has small dosage, high catalytic efficiency, reaction condition gentle
And solvent range of choice it is extensive the features such as, in recent years in terms of the structural modification of pharmaceutical synthesis field and polymeric carrier material
Be used widely (synthesis of Sheng Yongli, Zhu Zhengfang .4-dimethylamino naphthyridine and application study chemistry worlds, 1997,38
(10):528-529.) (the organic reaction synthesis chemistry of synthesis and its catalysis of Liao Lianan, Guo Qizhen .4- dimethylamino naphthyridines,
1995,3(3):215-221) (Liu Yaowu, gold pass mountain .DMAP and catalyze and synthesize resveratrol trinocotinate Suzhou University journal,
2010,25(2):39-41.) (Liang Ya, the high magnificent .DMAP of element catalyze and synthesize the research Tianjin science and engineering of three-O-acetylation erythromycin
Institute's journal, 2001,17 (2):18-20.).
However, DMAP be it is a kind of there is high toxicity and irritating material, to eyes, skin, alimentary canal, respiratory system and
Nervous system etc. can produce harm, the median lethal dose of its mouse mainline for 56mg/kg (https:// www.spectrumchemical.com/MSDS/TCI-D1450.pdf), seem so it is removed completely from synthetic product
It is particularly important.
Purification process mainly utilizes the alkaline and water miscible features of DMAP at present, is dialysed using pickling-WATER-WASHING METHOD or water
Method is removed.However, limitation be present in the above method, because in the synthesis of high polymer material, it is difficult to will be unreacted poly-
Compound separates from product and purifies removing, thus in synthesis the excessive mode of generally use acylating agent so that polymer as far as possible
Reaction is complete, and this, which is inevitably resulted in, is mixed with a large amount of DMAP- acylating agents intermediates in product.The property of this intermediate and DMAP is deposited
It is stronger in larger difference, usual hydrophobicity.If product is amphipathy macromolecule material, it is easily wrapped in aqueous environments
Enter in the micelle inner core that amphipathy macromolecule is self-assembly of, and reduce the purification efficiency of pickling or dialysis.
Cholesterol is animal tissue and the essential important substance of cell, because it has stronger lipophilicity, in recent years
To be used for frequently as hydrophobic grouping the structural modification of amphipathy macromolecule material.Xu etc. is with poly glycol monomethyl ether and cholesterol chlorine
Methyl esters (CHM) is raw material, reacts at room temperature and polyethylene glycol 2000 monomethyl ether-cholesterol carbonic ester (mPEG is made2000-CHMC)。(XU
H,DENG Y,CHEN D,et al.Esterase-catalyzed dePEGylation of pH-sensitive
vesicles modified with cleavable PEG-lipid derivatives.Journal of controlled
release,2008,130(3):238-245.) Song etc. obtains terminal graft using PLURONICS F87 (P188) and CHM as raw material
The P188- cholesterol carbonic ester (P188-CHMC) of cholesterol.(Song Y,Tian Q,Huang Z,et al.Self-
assembled micelles of novel amphiphilic copolymer cholesterol-coupled
F68containing cabazitaxel as a drug delivery system.International journal of
nanomedicine,2014,9:2307-2317.) above-mentioned reaction can be produced among a large amount of DMAP- cholesterol for being difficult to remove
Body.
Therefore, for being grafted the problem of DMAP purification efficiencies are relatively low in the amphipathy macromolecule material of cholesterol, it is badly in need of out
It is efficient to send out a kind of, the simple new method for removing DMAP.Since there is larger difference in DMAP- acylating agents intermediate and DMAP property
It is different, then intermediate is removed into a kind of effective ways of can yet be regarded as using this characteristic.
The content of the invention
Based on above-mentioned basis, using property of the DMAP- cholesterol intermediate insoluble in toluene, we devise one kind and are directed to
New method --- the toluene precipitation method that DMAP is removed in cholesterol amphipathy macromolecule material are grafted, and using the method DMAP's
Residual rate is less than 0.5%, and purity is up to more than 90%.
Removal DMAP of the present invention specific preparation method is as follows：
(1) grafting cholesterol Amphiphatic high polymer material is taken to be dissolved in toluene, ultrasound makes its molten after being heated to 25~100 DEG C
Solution.
(2) rotating speed is used to centrifuge DMAP- cholesterol intermediate and grafting cholesterol both sexes for 1000~15000rpm
The toluene solution of high polymer material.
(3) toluene solution layer is taken, sterling is produced after toluene is removed under reduced pressure.
Wherein, in described grafting cholesterol amphipathy macromolecule material, participate in reaction cholesterol can be cholesterol and
Its derivative such as cholesterol, cholesterol chloromethyl ester, Cholesteryl hemisuccinate, cholesterol acetate, cholesterol ester c etc., parent
Aqueous high molecular grafting materials cholesterol, its connecting key can be ester bond, carbonic acid ester bond, amido link, ehter bond, disulfide bond etc..
It is grafted in cholesterol amphipathy macromolecule material, the high polymer material for connecting cholesterol contains hydroxyl, amino, sulfydryl
Functional group, it can be polyethylene glycol and its derivative, polyaminoacid-polyethyleneglycol block copolymer, polyglycereol and its derivative
Thing, poly sialic acid or poloxamer, described polyethylene glycol and its derivative are 4- arms polyethylene glycol, 8- arms polyethylene glycol, gathered
Glycol monoethyl ether, folic acid-polyethylene glycol, TPGS, DSPE-poly-
Ethylene glycol, double nutmegs phosphatidyl-ethanolamine-polyethylene glycol, DPPE-polyethylene glycol, lauroyl phosphatide
Acyl monoethanolamine-polyethylene glycol, polycaprolactone-polyethylene glycol block copolymer, polylactic acid-polyethylene glycol block copolymer, molecule
Measure as 400~50000；Described polyaminoacid-polyethyleneglycol block copolymer is polyglutamic acid-polyethylene glycol block copolymerization
Thing, polyhistidyl-polyethyleneglycol block copolymer, polylysine-polyethyleneglycol block copolymer, poly-aspartate-poly- second two
Alcohol block copolymer, molecular weight are 400~50000；Polyglycereol and its derivative are phosphatidyl glycerol, distearyl acyl group phosphatidyl
Monoethanolamine-polyglycereol, molecular weight are 400~50000；Poly sialic acid, molecular weight are 400~50000；Poloxamer, Ru Boluo
Husky nurse 188, poloxamer188, poloxamer 237, Pluronic/Lutrol F 108 etc., molecular weight are 400~30000.
Described grafting cholesterol amphipathy macromolecule material can be PLURONICS F87-cholesterol carbonic ester, Bo Luosha
Nurse 407- cholesterol carbonic ester, poloxamer 237- cholesterol carbonic ester, Pluronic/Lutrol F 108-cholesterol carbonic ester, polyethylene glycol
400 VE-succinates-cholesterol carbonic ester, polyethylene glycol 1000 vitamin E succinic acid ester-cholesterol carbonic ester, poly- second
2000 VE-succinates of glycol-cholesterol carbonic ester, Macrogol 6000 VE-succinate-cholesterol carbonic acid
Ester, cetomacrogol 1000 monomethyl ether cholesterol hemisuccinic acid ester, polyethylene glycol 2000 monomethyl ether cholesterol hemisuccinic acid ester, poly- second
The monomethyl ether cholesterol hemisuccinic acid ester of glycol 10000, PEG 20000 monomethyl ether cholesterol hemisuccinic acid ester.
In the above method, it is preferable that heating-up temperature is 50~60 DEG C, and rotating speed is 5000~10000rpm, during vacuum drying
Between be 24~48h.
The advantage of the invention is that：(1) DMAP in the amphipathy macromolecule material of a variety of grafting cholesterol can be used for go
Remove；(2) the toluene precipitation method are simple to operate, purification efficiency is high, result is controllable, are advantageous to industrialized production.
Specific embodiment：
The present invention is expanded on further with reference to embodiment, these embodiments are merely to illustrate the present invention rather than limitation
The scope of the present invention.
Embodiment 1：The foundation of DMAP standard curves
DMAP 10.0mg are weighed in 10mL volumetric flasks, absolute ethyl alcohol is diluted to scale, shakes up, and obtains DMAP storing solutions
1.0mg·mL-1.Taking above-mentioned DMAP storing solutions 0.5mL, absolute ethyl alcohol is diluted to scale, shakes up, and obtains concentration into 50mL volumetric flasks
For 10.0 μ gmL-1DMAP solution.The above-mentioned μ gmL of DMAP 10.0 are pipetted respectively-1Solution 0.5,1.0,3.0,5.0,7.0,
9.0mL is diluted to scale in 10mL volumetric flasks, with absolute ethyl alcohol, shakes up, obtain concentration be respectively 0.5,1.0,3.0,5.0,
7.0、9.0μg·mL-1DMAP solution, using absolute ethyl alcohol as reference, at ultraviolet-visible spectrophotometer 281nm determine
Absorbance, and draw standard curve.
Linear regression is carried out to absorbance (A) with concentration (C), obtains regression equation A=0.1645C+0.0059 (R=
0.9999), show DMAP in 0.5~9.0 μ gmL-1Linear relationship is good in concentration range.
Embodiment 2：The dissolubility of different materials and DMAP intermediates in different solvents
Weigh respectively PLURONICS F87-cholesterol carbonic ester, PEG20000 monomethyl ether cholesterol hemisuccinic acid ester,
PEG20000 monomethyl ether cholesterol hemisuccinic acid ester and DMAP 5.0mg in cillin bottle, be separately added into acetonitrile, ether,
Hexamethylene, ethyl acetate, acetonitrile and toluene, the dissolubility of observation different materials and DMAP in multi-solvents, wherein "/" represent
Solvable, " ↓ " represents insoluble, the results are shown in Table 1.
The dissolubility of the different materials of table 1 and DMAP intermediates in a solvent
As seen from the above table, PLURONICS F87-cholesterol carbonic ester, PEG20000 monomethyl ether cholesterol hemisuccinic acid
Ester, PEG20000 monomethyl ether cholesterol hemisuccinic acid ester and DMAP intermediates are dissolved state in ethanol and acetonitrile,
It is insoluble in ether, hexamethylene and ethyl acetate, the phenomenon that material dissolves, DMAP is insoluble only can be observed in toluene, because
Solvent of this selection toluene as DMAP intermediates of going out.
Embodiment 3：PLURONICS F87-synthesis of cholesterol carbonic ester (P188-CHMC) and DMAP removal
Take 0.04mmol PLURONICS F87s (P188) to be placed in closed container, 0.02mmol DMAP are added under condition of nitrogen gas
With 0.14mmol triethylamines (TEA), 0.24mmol CHM dichloromethane (DCM) solution 16mL is slowly added dropwise, is stirred in ice-water bath
Mix after mixing 30min back flow reaction 36h under the conditions of 45 DEG C.Question response terminates rear removal of solvent under reduced pressure, precipitates in vain through ice ether
Color wax, then with ether washing precipitate 3 times (removing unreacted CHM) repeatedly, you can it is thick to obtain powdered P188-CHMC
Product.
(1) pickling-saturated sodium-chloride washes-WATER-WASHING METHOD：The P188-CHMC crude products 50mg for removing CHM is weighed in cillin bottle
In, distilled water is added, is extracted with DCM, then respectively with 100mmolL-1Hydrochloric acid, saturated sodium-chloride and frozen water are washed 3 times, and decompression removes
Go after solvent to precipitate through ice ether to produce P188-CHMC sterlings, vacuum drying 24h removes residual solvent.
(2) cation exchange resin processes：The sodium form cationic ion-exchange resin after 2.4mL (humid volume) processing is measured, is placed in
In 2.5mL syringes, 1000rpm centrifugation 5min, centrifugate is discarded, it is micro- to obtain 1.8mL (dry volume) sodium form cationic ion-exchange resin
Post is stand-by.2 parts of removal CHM P188-CHMC crude products 50mg is weighed in cillin bottle, is separately added into 15mL water and 50% ethanol
It is prepared into the P188-CHMC aqueous solution and 50% ethanol solution.The P188-CHMC solution of different solvents is splined on (processed
) sodium form resin cation microtrabeculae top, 2000rpm centrifugation 4min, eluent is collected, it is continuous to elute 3 times, merge eluent and remove
Go solvent to be precipitated through ice ether and produce P188-CHMC, vacuum drying 24h removes residual solvent.
(3) dialysis desivac：4 parts of removal CHM P188-CHMC crude products 50mg is weighed in cillin bottle, is dissolved in 8mL water
In, it is placed in the bag filter that molecular cut off is 5000Da.Respectively using pH 3.8,4.8,6.8 water and absolute ethyl alcohol as dialysis
Medium (volume 2L) is dialysed at room temperature, is changed dialyzate in 2,6,12,24 and 36h, is taken out bag filter after 48h, in bag
Liquid crosses 0.45 μm of filter membrane, and filtrate is lyophilized.
(4) the toluene precipitation method：The P188-CHMC 50mg for removing unreacted CHM are weighed in 10mL centrifuge tubes, add first
Benzole soln 8mL, after heating ultrasonic dissolution product, 10000rpm centrifugation 10min, undissolved DMAP-CHM intermediates are removed, together
Toluene layer is centrifuged again 1 time under the conditions of sample, be removed under reduced pressure after toluene and be settled out product through ice ether, vacuum drying 24h removes residual
Stay solvent.
By above-mentioned four kinds of methods purifying products therefrom P188-CHMC be dissolved in the absolute ethyl alcohol of certain volume, in it is ultraviolet-
Absorbance is determined at visible spectrophotometer 281nm, DMAP contents are calculated according to standard curve, and DMAP is calculated according to formula 1
Remaining rate, as a result as shown in table 1.
DMAP remnants' rate (%)=M/M0× 100% (1)
Wherein, M is the quality containing DMAP in every milligram of P188-CHMC after purification；M0For every milligram of P188- before purification
Quality containing DMAP in CHMC.
The distinct methods of table 2 purify P188-CHMC DMAP remnants' rate (n=3)
As shown in Table 1, in four kinds of purification process, pickling-saturation NaCl- WATER-WASHING METHODs and the cation tree using water as solvent
Fat method can not effectively remove the DMAP in P188-CHMC, and DMAP remnants' rate is respectively 44.33 ± 15.28% and 55.86 ±
2.57%, and pickling-saturated sodium-chloride wash-WATER-WASHING METHOD fluctuation it is larger, it is as a result not parallel.With the resin cation of 50% ethanol
Method, absolute ethyl alcohol can more effectively remove DMAP, DMAP residuals for the dialysis desivac and the toluene precipitation method of dialysis medium
Rate is less than 5%.Wherein, DMAP remnants' rate of the toluene precipitation method is only 0.20 ± 0.03%, can substantially completely remove DMAP.
Embodiment 4：Poloxamer188-synthesis of cholesterol carbonic ester (P407-CHMC) and DMAP removal
Take 0.04mmol poloxamer188s (P407) to be placed in closed container, 0.02mmol DMAP are added under condition of nitrogen gas
And 0.14mmolTEA, 0.24mmol CHM DCM solution 16mL are slowly added dropwise, 45 DEG C of bars after 30min are stirred in ice-water bath
Back flow reaction 36h under part.Question response terminates rear removal of solvent under reduced pressure, and White waxy thing is precipitated to obtain through ice ether, then anti-with ether
Multiple washing precipitate 3 times (removing unreacted CHM), you can obtain powdered P407-CHMC crude products.
The P407-CHMC 50mg for removing unreacted CHM are weighed in 10mL centrifuge tubes, add toluene solution 8mL, heating
Ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, to first under similarity condition
Benzene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces P407-CHMC sterlings, and vacuum drying 24h removes residual
Solvent.
Using method measure in embodiment 2 through the toluene precipitation method remaining rates of DMAP after purification, measured value for 0.35 ±
0.01%, DMAP can be substantially completely removed, and purity is 95.3%.
Embodiment 5：The synthesis of poloxamer 237- cholesterol carbonic ester (P237-CHMC) and DMAP removal
Take 0.04mmol poloxamers 237 (P237) to be placed in closed container, 0.02mmol DMAP are added under condition of nitrogen gas
And 0.14mmolTEA, 0.24mmol CHM DCM solution 16mL are slowly added dropwise, 45 DEG C of bars after 30min are stirred in ice-water bath
Back flow reaction 36h under part.Question response terminates rear removal of solvent under reduced pressure, and White waxy thing is precipitated to obtain through ice ether, then anti-with ether
Multiple washing precipitate 3 times (removing unreacted CHM), you can obtain powdered P237-CHMC crude products.
The P237-CHMC 50mg for removing unreacted CHM are weighed in 10mL centrifuge tubes, add toluene solution 8mL, heating
Ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, to first under similarity condition
Benzene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces P188-CHMC sterlings, and vacuum drying 24h removes residual
Solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP residual volumes after purification, measured value is 0.28 ±
0.004%, DMAP can be substantially completely removed, and purity is 92.8%.
Embodiment 6：Pluronic/Lutrol F 108-synthesis of cholesterol carbonic ester (P338-CHMC) and DMAP removal
Take 0.04mmol Pluronic/Lutrol F 108s (P338) to be placed in closed container, 0.02mmol DMAP are added under condition of nitrogen gas
And 0.14mmolTEA, 0.24mmol CHM DCM solution 16mL are slowly added dropwise, 45 DEG C of bars after 30min are stirred in ice-water bath
Back flow reaction 36h under part.Question response terminates rear removal of solvent under reduced pressure, and White waxy thing is precipitated to obtain through ice ether, then anti-with ether
Multiple washing precipitate 3 times (removing unreacted CHM), you can obtain powdered P338-CHMC crude products.
The P338-CHMC 50mg for removing unreacted CHM are weighed in 10mL centrifuge tubes, add toluene solution 8mL, heating
Ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, to first under similarity condition
Benzene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces P338-CHMC sterlings, and vacuum drying 24h removes residual
Solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.33 ±
0.02%, DMAP can be substantially completely removed, and purity is 97.8%.
Embodiment 7：Polyethylene glycol 400 VE-succinate-cholesterol carbonic ester (TPGS400- CHMC) synthesis and
DMAP removal
Take 0.66mmol polyethylene glycol 400 VE-succinates (TPGS400) be placed in closed container, under condition of nitrogen gas
41.5mg4- lutidines (DMAP) and 0.99mmol triethylamines (TEA) are added, the chloromethyl ester of cholesterol containing 1mmol is slowly added dropwise
(CHM) dichloromethane (DCM) solution 20mL, back flow reaction 24h, question response 25 DEG C at are stirred after 10min in ice-water bath
Removal of solvent under reduced pressure after end.
Weigh the TPGS for removing unreacted CHM400- CHMC 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces TPGS400- CHMC sterlings, vacuum drying 24h are removed
Residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP residual volumes after purification, measured value is 0.21 ±
0.005%, DMAP can be substantially completely removed, and purity is 94.8%.
Embodiment 8：Polyethylene glycol 1000 vitamin E succinic acid ester-cholesterol carbonic ester (TPGS1000- CHMC) synthesis
And DMAP purifying
Take 0.66mmol polyethylene glycol 1000 vitamin E succinic acid esters (TPGS1000) be placed in closed container, condition of nitrogen gas
Lower addition 41.5mg4- lutidines (DMAP) and 0.99mmol triethylamines (TEA), are slowly added dropwise the chloromethane of cholesterol containing 1mmol
Dichloromethane (DCM) solution 20mL of ester (CHM), back flow reaction 24h at 25 DEG C is stirred after 10min in ice-water bath, is treated anti-
Removal of solvent under reduced pressure after should terminating.
Weigh the TPGS for removing unreacted CHM1000- CHMC 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP-CHM intermediates of 2000rpm centrifugation 10min removings, to toluene under similarity condition
Layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces TPGS1000- CHMC sterlings, vacuum drying 36h remove residual
Stay solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.25 ±
0.01%, DMAP can be substantially completely removed, and purity is 98.6%.
Embodiment 9：Polyethylene glycol 2000 VE-succinate-cholesterol carbonic ester (TPGS2000- CHMC) synthesis
And DMAP purifying
Take 0.66mmol polyethylene glycol 2000 VE-succinates (TPGS2000) be placed in closed container, condition of nitrogen gas
Lower addition 41.5mg4- lutidines (DMAP) and 0.99mmol triethylamines (TEA), are slowly added dropwise the chloromethane of cholesterol containing 1mmol
Dichloromethane (DCM) solution 20mL of ester (CHM), back flow reaction 24h at 25 DEG C is stirred after 10min in ice-water bath, is treated anti-
Removal of solvent under reduced pressure after should terminating.
Weigh the TPGS for removing unreacted CHM2000- CHMC 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces TPGS2000- CHMC sterlings, vacuum drying 36h are removed
Remove residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.22 ±
0.005%, DMAP can be substantially completely removed, and purity is 94.7%.
Embodiment 10：Macrogol 6000 VE-succinate-cholesterol carbonic ester (TPGS6000- CHMC) synthesis
And DMAP purifying
Take 0.66mmol Macrogol 6000 VE-succinates (TPGS6000) be placed in closed container, condition of nitrogen gas
Lower addition 41.5mg4- lutidines (DMAP) and 0.99mmol triethylamines (TEA), are slowly added dropwise the chloromethane of cholesterol containing 1mmol
Dichloromethane (DCM) solution 20mL of ester (CHM), back flow reaction 24h at 25 DEG C is stirred after 10min in ice-water bath, is treated anti-
Removal of solvent under reduced pressure after should terminating.
Weigh the TPGS for removing unreacted CHM6000- CHMC 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces P188-CHMC sterlings, and vacuum drying 36h removes residual
Stay solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.31 ±
0.01%, DMAP can be substantially completely removed, and purity is 94.3%.
Embodiment 11：Cetomacrogol 1000 monomethyl ether cholesterol hemisuccinic acid ester (mPEG1000- CHS) synthesis and DMAP
Purifying
Take 0.66mmol cetomacrogol 1000 monomethyl ethers (mPEG1000) be placed in closed container, added under condition of nitrogen gas
41.5mg 4- lutidines (DMAP) and 142.9 μ L triethylamines (TEA), be slowly added dropwise Cholesteryl hemisuccinate (CHS,
Chloroformic solution 20mL 1mmol), is stirred after 10min that back flow reaction 12h, question response subtract after terminating at 60 DEG C in ice-water bath
Pressure removes solvent.
Weigh the mPEG for removing unreacted CHS1000- CHS 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces mPEG1000- CHS sterlings, vacuum drying 36h are removed
Residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.38 ±
0.02%, DMAP can be substantially completely removed, and purity is 96.1%.
Embodiment 12：Polyethylene glycol 2000 monomethyl ether cholesterol hemisuccinic acid ester (mPEG2000- CHS) synthesis and DMAP
Purifying
Take 0.66mmol polyethylene glycol 2000 monomethyl ethers (mPEG2000) be placed in closed container, added under condition of nitrogen gas
41.5mg 4- lutidines (DMAP) and 142.9 μ L triethylamines (TEA), be slowly added dropwise Cholesteryl hemisuccinate (CHS,
Chloroformic solution 20mL 1mmol), is stirred after 10min that back flow reaction 12h, question response subtract after terminating at 60 DEG C in ice-water bath
Pressure removes solvent.
Weigh the mPEG for removing unreacted CHS1000- CHS 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces mPEG1000- CHS sterlings, vacuum drying 48h are removed
Residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.38 ±
0.01%, DMAP can be substantially completely removed, and purity is 94.9%.
Embodiment 13：PEG20000 monomethyl ether cholesterol hemisuccinic acid ester (mPEG10000- CHS) synthesis and DMAP
Purifying
Take 0.66mmol PEG20000 monomethyl ethers (mPEG10000) be placed in closed container, added under condition of nitrogen gas
41.5mg 4- lutidines (DMAP) and 142.9 μ L triethylamines (TEA), be slowly added dropwise Cholesteryl hemisuccinate (CHS,
Chloroformic solution 20mL 1mmol), is stirred after 10min that back flow reaction 12h, question response subtract after terminating at 60 DEG C in ice-water bath
Pressure removes solvent.
Weigh the mPEG for removing unreacted CHS10000- CHS 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces mPEG10000- CHS sterlings, vacuum drying 48h are removed
Residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.29 ±
0.01%, DMAP can be substantially completely removed, and purity is 97.1%.
Embodiment 14：PEG 20000 monomethyl ether cholesterol hemisuccinic acid ester (mPEG20000- CHS) synthesis and DMAP
Purifying
Take 0.66mmol PEG 20000 monomethyl ethers (mPEG20000) be placed in closed container, added under condition of nitrogen gas
41.5mg 4- lutidines (DMAP) and 142.9 μ L triethylamines (TEA), be slowly added dropwise Cholesteryl hemisuccinate (CHS,
Chloroformic solution 20mL 1mmol), is stirred after 10min that back flow reaction 12h, question response subtract after terminating at 60 DEG C in ice-water bath
Pressure removes solvent.
Weigh the mPEG for removing unreacted CHS20000- CHS 50mg add toluene solution 8mL, added in 10mL centrifuge tubes
Hot ultrasound dissolves product, the undissolved DMAP- cholesterol intermediate of 10000rpm centrifugation 10min removings, right under similarity condition
Toluene layer centrifuges 1 time again, is removed under reduced pressure after toluene to precipitate through ice ether and produces mPEG20000- CHS sterlings, vacuum drying 48h are removed
Residual solvent.
Using method measure in embodiment 2 through toluene precipitation method DMAP remnants' rate after purification, measured value is 0.25 ±
0.01%, DMAP can be substantially completely removed, and purity is 91.3%.
Claims (10)
1. it is grafted the minimizing technology of DMAP in cholesterol amphipathy macromolecule material, it is characterised in that using the toluene precipitation method.
2. the minimizing technology of DMAP in cholesterol amphipathy macromolecule material is grafted as claimed in claim 1, it is characterised in that
Comprise the following steps：
(1) grafting cholesterol amphipathy macromolecule material is taken to be dissolved in toluene, ultrasound makes its dissolving after being heated to 25~100 DEG C；
(2) rotating speed is used to centrifuge DMAP- cholesterol intermediate and the grafting amphipathic height of cholesterol for 1000~15000rpm
The toluene solution of molecular material；
(3) toluene solution layer is taken, grafting cholesterol amphipathy macromolecule material is produced after toluene is removed under reduced pressure.
3. DMAP minimizing technology, its feature in grafting cholesterol amphipathy macromolecule material according to claim 1 or 2
For in described grafting cholesterol amphipathy macromolecule material, its connecting key is ester bond, carbonic acid ester bond, amido link, ehter bond or two
Sulfide linkage.
4. DMAP removal side in the grafting cholesterol amphipathy macromolecule material according to claim 1-3 any one
Method, it is characterized in that, it can be cholesterol, cholesterol chloromethyl ester, cholesterol hemisuccinic acid to participate in the cholesterol of reaction and its derivative
Ester, cholesterol acetate or cholesterol ester c.
5. DMAP removal side in the grafting cholesterol amphipathy macromolecule material according to claim 1-4 any one
Method, it is characterized in that, grafting cholesterol amphipathy macromolecule material contains the functional group of hydroxyl, amino, sulfydryl, including：Poly- second two
Alcohol and its derivative, polyaminoacid-polyethyleneglycol block copolymer, polyglycereol and its derivative, poly sialic acid or poloxamer.
6. DMAP minimizing technology in grafting cholesterol amphipathy macromolecule material according to claim 5, it is characterized in that,
Described polyethylene glycol and its derivative are 4- arms polyethylene glycol, 8- arms polyethylene glycol, poly glycol monomethyl ether, folic acid-poly- second
Glycol, TPGS, DSPE-PEG, double nutmeg phosphatidyl second
Hydramine-polyethylene glycol, DPPE-polyethylene glycol, lauroyl phosphatidyl-ethanolamine-polyethylene glycol, gather oneself
Lactone-polyethyleneglycol block copolymer, polylactic acid-polyethylene glycol block copolymer, molecular weight are 400~50000；Described is poly-
Amino acid-polyethyleneglycol block copolymer is polyglutamic acid-polyethyleneglycol block copolymer, polyhistidyl-polyethylene glycol block
Copolymer, polylysine-polyethyleneglycol block copolymer, poly-aspartate-polyethyleneglycol block copolymer, molecular weight 400
~50000；Described polyglycereol and its derivative are phosphatidyl glycerol, DSPE-polyglycereol, molecule
Measure as 400~50000；Described poly sialic acid molecular weight is 400~50000；Described poloxamer, molecular weight be 400~
50000。
7. DMAP minimizing technology in grafting cholesterol amphipathy macromolecule material according to claim 5, it is characterized in that,
It is PLURONICS F87-cholesterol carbonic ester, poloxamer188-cholesterol carbonic acid to be grafted cholesterol amphipathy macromolecule material
Ester, poloxamer 237- cholesterol carbonic ester, Pluronic/Lutrol F 108-cholesterol carbonic ester, polyethylene glycol 400 VE succinic acid
Ester-cholesterol carbonic ester, polyethylene glycol 1000 vitamin E succinic acid ester-cholesterol carbonic ester, polyethylene glycol 2000 vitamin E
Succinate-cholesterol carbonic ester, Macrogol 6000 VE-succinate-cholesterol carbonic ester, cetomacrogol 1000 list
Methyl ether Cholesteryl hemisuccinate, polyethylene glycol 2000 monomethyl ether cholesterol hemisuccinic acid ester, PEG20000 monomethyl ether courage
Sterol hemisuccinic acid ester or PEG 20000 monomethyl ether cholesterol hemisuccinic acid ester.
8. DMAP minimizing technology in grafting cholesterol amphipathy macromolecule material according to claim 1, it is characterized in that,
Heating-up temperature is 50~60 DEG C in step (1).
9. DMAP minimizing technology in grafting cholesterol amphipathy macromolecule material according to claim 1, it is characterized in that,
5000~10000rpm of rotating speed in step (2).
10. DMAP minimizing technology, its feature in grafting cholesterol amphipathy macromolecule material according to claim 1
For 4~72h of vacuum drying time, preferably 24~48h in step (3).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610550318.6A CN107619473B (en) | 2016-07-13 | 2016-07-13 | Method for removing DMAP (dimethyl acetamide) in synthesis of grafted cholesterol amphiphilic polymer material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610550318.6A CN107619473B (en) | 2016-07-13 | 2016-07-13 | Method for removing DMAP (dimethyl acetamide) in synthesis of grafted cholesterol amphiphilic polymer material |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107619473A true CN107619473A (en) | 2018-01-23 |
| CN107619473B CN107619473B (en) | 2020-04-10 |
Family
ID=61087058
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610550318.6A Active CN107619473B (en) | 2016-07-13 | 2016-07-13 | Method for removing DMAP (dimethyl acetamide) in synthesis of grafted cholesterol amphiphilic polymer material |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107619473B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108409823A (en) * | 2018-03-12 | 2018-08-17 | 山西潞安煤基合成油有限公司 | A kind of cholesterine lipoid substance and preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068701A (en) * | 2011-01-18 | 2011-05-25 | 沈阳药科大学 | Application of cleavable polyethylene glycol (PEG) lipid derivative to preparation |
| CN104844790A (en) * | 2014-02-19 | 2015-08-19 | 沈阳药科大学 | Tocopheryl polyethylene glycol succinate-cholesterol carbonate, and preparation method and application thereof |
-
2016
- 2016-07-13 CN CN201610550318.6A patent/CN107619473B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102068701A (en) * | 2011-01-18 | 2011-05-25 | 沈阳药科大学 | Application of cleavable polyethylene glycol (PEG) lipid derivative to preparation |
| CN104844790A (en) * | 2014-02-19 | 2015-08-19 | 沈阳药科大学 | Tocopheryl polyethylene glycol succinate-cholesterol carbonate, and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| CAN YANG ZHANG ET AL.: ""self-assembled micelles based on pH-sensitive PAE-g-MPEG-cholesterol block copolymer for anticancer drug delivery"", 《INTERNATIONAL JOURNAL OF NANOMEDICINE》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108409823A (en) * | 2018-03-12 | 2018-08-17 | 山西潞安煤基合成油有限公司 | A kind of cholesterine lipoid substance and preparation method and application |
| CN108409823B (en) * | 2018-03-12 | 2021-02-26 | 山西潞安煤基合成油有限公司 | Cholesterol lipid compound and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107619473B (en) | 2020-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dai et al. | Lignin nanoparticle as a novel green carrier for the efficient delivery of resveratrol | |
| CN101327328B (en) | Dendrimers targeting nano particle and preparation and application thereof | |
| CN102604114B (en) | Star-shaped cationic polymer containing dendriform polylysine element and preparation method thereof | |
| CN106432746B (en) | A kind of amphipathic hydroxyethyl starch coupling copolymer of poly lactic acid and its preparation method and application | |
| CN1698901A (en) | Chitosan or its derivative as drug carrier for carrying red sage root extract | |
| CN102260356B (en) | Chitosan derivative used as gene vector, and preparation method and application thereof | |
| CN113292608B (en) | Exosome drug delivery system and preparation method and application thereof | |
| Sun et al. | Integrated Janus nanofibers enabled by a co-shell solvent for enhancing icariin delivery efficiency | |
| CN107158410B (en) | Folic acid-chitosan-Cy 7 polymer with tumor targeting property and preparation method thereof | |
| CN102139112A (en) | Drug carrier raw material and preparation method and application thereof | |
| CN107213467A (en) | A kind of preparation method of phospholipid complexes of curcumin | |
| Esmaeili et al. | Synthesis of a novel structure for the oral delivery of insulin and the study of its effect on diabetic rats | |
| Xia et al. | Biodegradable and self-fluorescent ditelluride-bridged mesoporous organosilica/polyethylene glycol-curcumin nanocomposite for dual-responsive drug delivery and enhanced therapy efficiency | |
| Pandey et al. | Development of biodegradable chitosan/graphene oxide nanocomposite via spray drying method for drug loading and delivery application | |
| Dubey et al. | Nanostructured lipid carriers of ivabradine hydrochloride: optimization, characterization and in-vivo estimation for management of stable angina | |
| CN101773471B (en) | Mitoxantrone targeting sustained-release long-circulating nanometer liposome and preparation method | |
| CN107619473A (en) | It is grafted the minimizing technology of DMAP in cholesterol amphipathy macromolecule materials synthesis | |
| CN108542895A (en) | Hyaluronic acid-curcumin-polyethylene glycol carrier preparation method and applications | |
| CN102772367B (en) | Preparation method of targeting composite nanoparticle | |
| CN105168230B (en) | A kind of cancer target prodrug and its nanometer formulation and preparation method with endosome escape function | |
| CN106667962A (en) | Charge conversion type nanometer drug carrier and preparation method thereof | |
| CN107286349A (en) | Polyethylene glycol polycaprolactone diblock copolymer and its preparation method and application | |
| Cheng et al. | Diblock copolymer glyco-nanomicelles constructed by a maltoheptaose-based amphiphile for reduction-and pH-mediated intracellular drug delivery | |
| CN104031268B (en) | Polyethyleneimine triblock copolymer and application thereof to gene carriers | |
| Gao et al. | Hydrotropic polymer-based paclitaxel-loaded self-assembled nanoparticles: preparation and biological evaluation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20190130 Address after: 110016 No. 103, cultural road, Shenhe District, Shenyang, Liaoning Applicant after: Shenyang Pharmaceutical University Applicant after: Guangzhou Zhigao Point Pharmaceutical Technology Co., Ltd. Address before: 110016 No. 103, cultural road, Shenhe District, Shenyang, Liaoning Applicant before: Shenyang Pharmaceutical University |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |