CN102343099A - Preparation method and application of folacin mediated tumor targeting adriamycin prodrugs - Google Patents
Preparation method and application of folacin mediated tumor targeting adriamycin prodrugs Download PDFInfo
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- CN102343099A CN102343099A CN 201110299651 CN201110299651A CN102343099A CN 102343099 A CN102343099 A CN 102343099A CN 201110299651 CN201110299651 CN 201110299651 CN 201110299651 A CN201110299651 A CN 201110299651A CN 102343099 A CN102343099 A CN 102343099A
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Abstract
The invention provides a preparation method and application of two types of adriamycin prodrugs. The two types of prodrugs with tumor active targeting and acid-sensitive initial burst release properties can be prepared by virtue of connection of hydrazone bonds and introduction of folacin. By utilizing the two types of prodrugs, the tumor resistance of adriamycin can be reinforced, the toxic and side effects of the medicament can be reduced, and the targeting property can be improved. Therefore, the preparation method provides a novel idea for the development of anti-tumor medicaments.
Description
Technical field
The present invention relates to the method for preparing and the application of two kinds of adriablastina prodrugs.
Background technology
Malignant tumor serious harm human health and existence are to cause one of main causes of death, and its sickness rate can be in any more always, have presented tangible rejuvenation trend at present.2000, World Health Organization's report, global cancer mortality example number surpasses 7,000,000, and wherein developed country accounts for 21%, and developing country accounts for 9%, and the death toll that tumor causes accounts for 12% of global death toll.Malignant tumor increases case about 2,000,000, dead 1,700,000 every year newly in the primary cause of death that China has become urban and rural residents.
Anthracycline antibiotics is one of clinical the most frequently used and effective antitumour medicine, and amycin is the representative medicine of this series antineoplastic medicament, early than from streptococcic metabolism of pigment product, obtaining the sixties in last century.Amycin is a kind of broad-spectrum chemotherapeutics, is usually used in treating breast carcinoma clinically, child's solid tumor, soft-tissue tumor, the malignant tumor such as lymphatic cancer of feeling sick.Clinical research is in recent years found, in the clinical practice of amycin, has occurred the drug resistance to tumor cell, and the cytotoxicity for normal structure has limited its further application simultaneously.The modal toxicity of this type of life-time service medicine is injury of myocardium, can bring out congestive heart failure.In view of above reason; People are continuing to the transformation and the modification of amycin always; Specialized transport carrier through will having tumor-targeting or can be combined with amycin by the molecule of tumor tissues specific recognition; Processing the adriablastina prodrug with tumor-targeting, is that present people are devoted to seek one of strategy that increases its drug effect and safety always.
At present, according to the design concept of prodrug, mainly concentrate on 13 carbonyl and 3 ' amino to the position of amycin structure of modification.Wherein, 13 carbonyl can be connected with the hydrazides group (modification group) of different molecular, the reaction of shrinking forms hydrazone key junctional complex, and the hydrazone key is more stable under neutral environment, and cleavable discharges the former medicine of amycin under the environment of meta-acid.Research shows, the growth of tumour cell metabolism is active, and pH value is starkly lower than normal structure on every side, so the formation of hydrazone key can bring into play acid-sensitive effect, has the effect of cancer target.
Different carrier molecules have different features, and itself and the bonded mode of former medicine or the characteristics that act on tumor tissues are difference to some extent.Polyethylene Glycol (PEG) can delay the speed that medicine is removed, and the half-life of prolong drug, the increase antitumor drug is accumulated tumor locus, and local concentration can reach 5-10 doubly, and then the effect of performance tumor tissues passive target.PEG can be connected through covalent bond with multiple medicine, thereby synthetic different macromolecule precursor medicament has been widely used in pharmaceutical preparation and the drug delivery system at present.
At present, the tumor cell folacin receptor receives extensive concern.Folacin receptor is the membrane glycoprotein that a kind of glycosylation phospholipid inositol connects, high expressed in many tumor cells, and in most of normal structures, express hardly.Therefore folic acid is connected with antitumor drug, can makes drug targeting to tumor cell, killing tumor cell optionally, and alleviate infringement to normal structure.
Summary of the invention
The purpose of this invention is to provide the method for preparing and the application of two kinds of adriablastina prodrugs; Amycin combined with Polyethylene Glycol, the folic acid modified respectively obtain two kinds and have acid-sensitive active adriablastina target precursor medicine; May strengthen the antitumor action of amycin; Reduce poisonous side effect of medicine; Improve targeting property, for the exploitation of antitumor drug provides new thinking.
Precursor medicine of the present invention has two kinds: 1. amycin combines the amycin precursor medicine that obtains with Polyethylene Glycol, folic acid.
2. amycin is connected the amycin prodrug that obtains with folic acid through aminocaproic acid.
The structural formula of amycin is following:
The synthetic route of folic acid-aminocaproic acid-amycin:
Concrete synthesis method is following:
Synthesizing of 1 aminocaproic acid ethyl ester
Add dehydrated alcohol in the there-necked flask, the cryosel water-bath is cooled to below-10 ℃, drips the thionyl chloride that heavily steams, and maintains the temperature at below 0 ℃.Drip and finish, room temperature is stirring 10h down, adds aminocaproic acid temperature rising reflux reaction 7h, and decompression steams excessive thionyl chloride and ethanol, and residue solidifies, and uses methanol: (v: v=5: 1) recrystallization gets white solid to petroleum ether.
Synthesizing of 2 folic acid aminocaproic acid ethyl esters
Precision takes by weighing folic acid and adds anhydrous dimethyl sulphoxide (DMSO), and stirring and evenly mixing adds NHS more successively, DCC, triethylamine, room temperature nitrogen lucifuge activation 6h; Add the aminocaproic acid ethyl ester then, reaction 12h.Filtering reaction by-product DCU uses acetone-ether sedimentation at last, and ether washes repeatedly, gets crude product.Cross post separate pure article.
Synthesizing of 3 folic acid aminocaproic acid hydrazides
Precision takes by weighing folic acid aminocaproic acid ethyl ester, is dissolved among the anhydrous DMSO, adds the hydrazine hydrate of 4mL then, and 50 ℃ of lucifuge reaction 6h use ethanol precipitation at last, and ether washes repeatedly, gets crude product.Cross post separate pure article.Synthesizing of 4 folic acid-aminocaproic acid-amycin
Precision takes by weighing folic acid aminocaproic acid hydrazides, is dissolved among the anhydrous DMSO.Fully the dissolving back adds the 75mg amycin, slowly drips the trifluoroacetic acid of 60 μ L, room temperature lucifuge reaction 12h; Use acetone-ether sedimentation at last, ether washes repeatedly, gets crude product.Cross post separate pure article.
The synthesis method of amycin-PEG-folic acid hydrazone key junctional complex is following:
1, HOOC-PEG-folic acid is synthetic
Folic acid is dissolved among the anhydrous DMSO, after nitrogen protection activates 30min in advance under the adding DCC/NHS room temperature, adds HOOC-PEG-NH
2, system in the presence of triethylamine in room temperature reaction 2h.Add water after reaction finishes pH transferred to 7.5, the extraction that adds methylene chloride in the reactant liquor, add diethyl ether precipitate product.
2, amycin-PEG-folic acid hydrazone key junctional complex
HOOC-PEG-folic acid and tert-butyl group oxygen carbonyl hydrazine (BOC-hyd) react in DMSO under the catalysis of DCC/NHS and obtain FA-PEG-hyd-BOC, in 20% hydrochloric acid, slough the BOC blocking group and obtain FA-PEG-hyd.FA-PEG-hyd is dissolved among the DMSO, adds amycin, lucifuge reaction 48h gets amycin-PEG-folic acid hydrazone key junctional complex (DOX-hyd-PEG-FA) in the presence of trifluoroacetic acid.
Concrete synthetic route is following:
Description of drawings
Fig. 1 be folic acid-aminocaproic acid-amycin (FA-AMA-DOX) in 24h to KB cells in vitro anti-tumor activity curve chart.
Fig. 2 be folic acid-aminocaproic acid-amycin (FA-AMA-DOX) in 24h to A549 cells in vitro anti-tumor activity curve chart.
Amycin in Fig. 1 and Fig. 2 (DOX), aminocaproic acid-amycin (AMA-DOX), folic acid-aminocaproic acid-amycin (FA-AMA-DOX) in 24h to the cytotoxicity of KB cell (A) and A549 cell (B).
Fig. 3 is that amycin-PEG-folic acid hydrazone key junctional complex (DOX-hyd-PEG-FA) is hatched behind the 48h A549 cells in vitro anti-tumor activity figure.
Fig. 4 is that amycin-PEG-folic acid hydrazone key junctional complex (DOX-hyd-PEG-FA) is hatched behind the 48h KB cells in vitro anti-tumor activity figure.
Free amycin (DOX) in Fig. 3 and Fig. 4; Amycin-PEG amido link junctional complex (DOX-ami-PEG); Amycin-PEG-folic acid amido link junctional complex (DOX-ami-PEG-FA); Amycin-PEG hydrazone key junctional complex (DOX-hyd-PEG); Amycin-PEG-folic acid hydrazone key junctional complex (DOX-hyd-PEG-FA) is hatched behind the 48h A549 cell (A), the cytotoxicity of KB cell (B).
The practical implementation method
The anti tumor activity in vitro research of adriablastina prodrug
1 purpose:
Through experiment in vitro, with the positive contrast of amycin, it is active to estimate the prodrug antineoplastic.
2 research methoies:
2.1 the prodrug vitro cytotoxicity is estimated
Various medicines to be measured are dissolved in (DMSO is lower than 0.1%) in the culture fluid that does not contain serum by setting concentration; Trypsinization with 0.25% grows to the negative A549 cell of expressing of KB cell, folic acid of the folic acid high expressed of exponential phase; Be inoculated in 96 orifice plates, density is 1 * 10
4Add 0.001 respectively after cultivating 24h; 0.01,0.1,1; The DOX of 10 μ M (being the concentration of amycin), FA-AMA-DOX, DOX-hyd-PEG-FA (culture fluid that does not wherein contain serum is a blank); Adding 20 μ L MTT continue to hatch 4h after hatching 24h/48h, discard culture fluid, with 150 μ L DMSO dissolve purple crystallizations again; Microplate reader reads the OD value under 570nm, draw the cell survival rate curve chart.
3 experimental results:
Experimental result is seen accompanying drawing.
Fig. 1 has shown folic acid-aminocaproic acid-amycin (FA-AMA-DOX) anti tumor activity in vitro.Compare with the junctional complex that is not connected folic acid, the anti tumor activity in vitro of FA-AMA-DOX demonstrates good antitumor activity on the KB of folacin receptor high expressed cell, and anti tumor activity in vitro is better than amycin; But for the negative A549 cell of cell surface folacin receptor, folate-mediated prodrug does not demonstrate ideal anti tumor activity in vitro.This result confirms that the tumor cell of FA-AMA-DOX pair cell surface folacin receptor high expressed has good in vitro and suppresses active.
Fig. 2 has shown the anti tumor activity in vitro of amycin-PEG-folic acid hydrazone key junctional complex (DOX-hyd-PEG-FA).Similar with FA-AMA-DOX, for the KB cell of folacin receptor high expressed, the introducing of folic acid can increase the absorption of tumor cell to prodrug, thereby improves drug level in the cell, increases the anti-tumor activity of prodrug.Compare with amycin-PEG-folic acid amido link junctional complex (DOX-ami-PEG-FA) simultaneously; Hydrazone key junctional complex is because the introducing of the acid-sensitive active hydrazone key of tool; Realized prominent release of the former medicine of amycin in tumor cell; Thereby increased the concentration of free amycin in the tumor cell, demonstrated certain anti tumor activity in vitro.
4 conclusions:
Visible by above cell experiment result; Folate-mediated amycin-Polyethylene Glycol hydrazone key macromole junctional complex and amycin-aminocaproic acid-folic acid micromolecule junctional complex can be enriched in the abundant tumor cell of folacin receptor through folacin receptor mediated approach targeting; Reach targeting; Improve curative effect; Reduce toxic and side effects, have good DEVELOPMENT PROSPECT.
Claims (3)
1. the method for preparing of folic acid-aminocaproic acid-adriablastina prodrug; It is characterized in that glycine and ethanol synthesis are obtained ethyl aminoacetate; Be connected through DCC/NHS catalysis with folic acid and obtain the folic acid ethyl aminoacetate, carry out ester exchange with hydrazine hydrate and obtain folic acid glycine hydrazides; Through the catalytic action of trifluoroacetic acid, the amycin introducing is obtained target compound folic acid-aminocaproic acid-amycin.
2. the method for preparing of amycin-PEG-folic acid hydrazone key junctional complex is characterized in that folic acid through DCC/NHS catalysis and HOOC-PEG-NH
2Connection obtains HOOC-PEG-FA, and HOOC-PEG-folic acid and tert-butyl group oxygen carbonyl hydrazine BOC-hyd react under the catalysis of DCC/NHS and obtain FA-PEG-hyd-BOC, slough the BOC blocking group and obtain FA-PEG-hyd; Add amycin, in the presence of trifluoroacetic acid amycin-PEG-folic acid hydrazone key junctional complex DOX-hyd-PEG-FA.
3. two kinds of adriablastina prodrug application in treatment folacin receptor high expressed tumor.
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| CN102766262A (en) * | 2012-07-20 | 2012-11-07 | 上海交通大学 | Preparation method for difunctional nanoparticle carrier and preparation method for difunctional nanoparticle preparation |
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| CN108452317A (en) * | 2018-03-29 | 2018-08-28 | 北京林业大学 | A kind of medicine-carried nano particles and preparation method thereof based on pectin/doxorubicin conjugates |
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- 2011-09-30 CN CN2011102996511A patent/CN102343099B/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 《Journal of Controlled Release》 20041124 Hyuk Sang Yoo et al. "Folate-receptor-targeted delivery of doxorubicin nano-aggregates stabilized by doxorubicin-PEG-folate conjugate" 第247－256页 2 第100卷, 第2期 * |
| 《中国药理学通报》 20091031 宦梦蕾等 "肿瘤靶向阿霉素前体药物的设计合成及其抗肿瘤活性研究" 第25卷 2 第25卷, * |
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| CN102766262B (en) * | 2012-07-20 | 2014-08-06 | 上海交通大学 | Preparation method for difunctional nanoparticle carrier and preparation method for difunctional nanoparticle preparation |
| CN102766262A (en) * | 2012-07-20 | 2012-11-07 | 上海交通大学 | Preparation method for difunctional nanoparticle carrier and preparation method for difunctional nanoparticle preparation |
| CN103381273B (en) * | 2013-05-29 | 2016-06-29 | 南方医科大学 | Amycin prodrug and preparation method thereof and injectable compositions |
| CN103381273A (en) * | 2013-05-29 | 2013-11-06 | 南方医科大学 | Doxorubicin prodrug, its preparation method and injectable composition |
| CN105055375B (en) * | 2015-07-15 | 2017-11-14 | 中国人民解放军第四军医大学 | Adriamycin and resveratrol with the effect of anti-multidrug resistance deliver nanoscale medicine delivery system altogether |
| CN105055375A (en) * | 2015-07-15 | 2015-11-18 | 中国人民解放军第四军医大学 | Amycin and resveratrol co-delivery nano drug delivery system with anti-multi-drug-resistance effect |
| CN105903031A (en) * | 2016-05-05 | 2016-08-31 | 上海交通大学 | Preparation method and application of drug controlled release nano-system sensitive to tumor microenvironment |
| CN106139159A (en) * | 2016-08-30 | 2016-11-23 | 四川大学 | The polyethyleneglycol derivative small-molecule drug conjugate polymer micelle sensitive for pH purposes in inflammation targeted delivery system |
| CN106692986A (en) * | 2016-12-09 | 2017-05-24 | 浙江医药高等专科学校 | Docetaxel clathrate compound as well as preparation method and application of docetaxel clathrate compound |
| CN106692986B (en) * | 2016-12-09 | 2019-11-19 | 浙江医药高等专科学校 | The preparation method and application of docetaxel inclusion compound, docetaxel inclusion compound |
| CN107098907A (en) * | 2017-06-13 | 2017-08-29 | 佛山科学技术学院 | A kind of bicyclic alcohols folate conjugate and its production and use |
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