CN102276813A

CN102276813A - Targeting polymer drug carrier containing folic acid and preparation method thereof

Info

Publication number: CN102276813A
Application number: CN 201110238131
Authority: CN
Inventors: 熊向源; 郭亮; 李资玲; 龚妍春; 李玉萍
Original assignee: Jiangxi Technology Normal College
Current assignee: Jiangxi Technology Normal College
Priority date: 2011-08-19
Filing date: 2011-08-19
Publication date: 2011-12-14
Anticipated expiration: 2031-08-19
Also published as: CN102276813B

Abstract

The invention discloses an amphiphilic block polymer namely polymer folic acid-polyoxyethylene-polyoxypropylene-polyoxyethylene-polylactic acid (FA-Pluronic-PLA) which contains a folic acid targeting group, polyoxyethylene-polyoxypropylene-polyoxyethylene as a hydrophilic chain segment and polylactide as a hydrophobic chain segment and has a novel chemical structure, and a preparation method of the compound.

Description

Contain folic acid target polymer drug carrier and preparation method thereof

Technical field

The invention belongs to biomedicine field, relate to a kind of folic acid target polymer drug carrier and preparation method thereof that contains.

Background technology

Amphipathy macromolecule, especially biocompatible amphipathy macromolecule (just containing two kinds of segmental polymers of wetting ability and hydrophobicity) is widely studied, because they can form the nanoparticle with various different shapes by the self aggregations such as hydrophobic interaction between hydrophobic segment in water, this character makes amphipathy macromolecule at drug delivery system very big application prospect be arranged, as controllable release system, target delivery systme etc.We know, present most of medicine (as cancer therapy drug) is hydrophobic, and is just water insoluble, is easy to be excreted by the intravital a series of rejections of people, as medicine resistancing action, enzyme degradation or the like, this has limited the validity of disease treatments such as cancer greatly.And the nanoparticle that amphipathy macromolecule forms can be used as pharmaceutical carrier; pharmaceutical pack is embedded in the hydrophobic core; the surface is by the protection of the hydrophilic layer of nanoparticle, just medicine can be transported to diseased region (as tumour etc.) like this, thereby plays the effect of effective treatment cancer.

The modification of target group is carried out on the surface of high molecular nanometer particles, can be improved the selectivity of drug conveying and the validity of disease treatment.Because the targeting type nanoparticle is transported to diseased regions such as tumour with the medicine orientation of embedding, so both can reduce medicine to Normocellular infringement, can reduce amount of drug because of improving utilization ratio of drug again, thereby alleviate the side effect that medicine produces human body.Therefore, the targeting type polymeric nanoparticle has very big application prospect at drug delivery system.

Folic acid is the necessary VITAMIN of cell (especially proliferative cell), participates in a carbon shift reaction of multiple pathways metabolism.The cell traffic of folic acid is by two kinds of transmembrane proteins, and promptly (folate receptor FR) finishes the folacin receptor of the reductibility folate carrier of low-affinity and high-affinity.Confirmed FR at present in the overexpression of kinds of tumor cells surface, and the expression in most healthy tissuess only limits to some and is difficult to enter sanguimotor epithelial cell teleblem.Just because of the characteristic that FR expresses, (folic acid FA) becomes the important molecule of drug targeting to tumour cell FR native ligand-folic acid, and folic acid has the high-affinity (K with folacin receptor _d=l * 10 ^-10MolL ^-1), reduced immunogenicity, be easy to modify, the little (M of volume _w=441.4), height chemical stability and biological stability, high tumour perviousness, easily combine with medicine, with advantages such as the consistency of organic and aqueous solvent and low costs, the research of folate-mediated cancer target is developed rapidly.

In recent years, both at home and abroad to the existing research more widely of the polymer drug delivery systme of folic acid target.The J. R. Baker Jr. seminar of U.S. University of Michigan in the finishing of polyamide-amide (PAMAM) dendrimer target group folic acid, the result shows that the KB cell of pharmaceutical carrier behind the modified with folic acid and surperficial overexpression folacin receptor has specificity to interact, thereby can improve result of treatment (Choi, the Y. of cancer therapy drug effectively Chemistry ﹠amp; Biology2005,12,35), still, the clinical application of dendritic macromole depends on the development of correlative studys such as controlled preparation, functionalization largely, and also there are problems such as difficulty is big, cost height in the controlled preparation of the monodispersity peptide class dendritic macromole of Present Attitude regularity.

And segmented copolymer can be realized effective controlledly synthesis by polymerization processs such as living polymerization, ring-opening polymerizations.Folic acid-polycaprolactone-polyethylene glycol (FA-PCL-PEO) segmented copolymer of synthetic embedding taxols such as Park has higher cytotoxicity (Park, E. K. than PCL-PEO segmented copolymer to cancer cells Journal of Controlled Release2005,109,158).So far, synthetic (Lee, the E. S. that have only minority reported in literature folic acid-polyethylene glycol-lactic acid (FA-PEO-PLA) segmented copolymer Journal Of Controlled Release2003,91,103-113).As for the FA-Pluronic-PLA segmented copolymer, had not yet to see reported in literature.

Summary of the invention

Purpose of the present invention just provide a kind of contain folic acid target group, by polyoxyethylene-oxypropylene-oxygen ethene (PEO-PPO-PEO, Pluronic) as hydrophilic segment, polylactide [poly (lactic acid), PLA] as the amphipathic nature block polymer folic acid-polyoxyethylene-poly-oxypropylene polyoxyethylene-poly(lactic acid) (FA-Pluronic-PLA) of hydrophobic segment, and the preparation method of this compound.

Folic acid target polymer drug carrier--the folic acid-polyoxyethylene-poly-oxypropylene polyoxyethylene-poly(lactic acid) (FA-Pluronic-PLA) that contains of the present invention, its chemical structural formula is as follows:

Figure 201110238131X100002DEST_PATH_IMAGE001

K, m, n represent lactic acid, oxygen ethene, oxypropylene number of repeating units respectively.

Folic acid target polymer drug carrier--the folic acid-polyoxyethylene-poly-oxypropylene polyoxyethylene-poly(lactic acid) (FA-Pluronic-PLA) that contains of the present invention, its synthetic route is as follows:

Figure 201110238131X100002DEST_PATH_IMAGE002

The folic acid target polymer drug carrier that contains of the present invention--folic acid-polyoxyethylene-poly-oxypropylene polyoxyethylene-poly(lactic acid) (FA-Pluronic-PLA) preparation method is:

1, FA-Pluronic-OH's is synthetic: with 0.38-0.49g folic acid (folic acid, FA) with 0.18g 1,3-dicyclohexylcarbodiimide (DCC) joins in the 35ml anhydrous dimethyl sulfoxide (DMSO), at room temperature stirred 10-15 hour, again with 10g, 0.79mmol polyoxyethylene-poly-oxypropylene polyoxyethylene (Pluronic) and 0.097g 4-Dimethylamino pyridine (DMAP) add wherein, at room temperature continue to stir 24-48 hour, reactant is centrifugal, getting supernatant dialysed 2-4 hour with anhydrous dimethyl sulfoxide, the molecular weight cut-off of dialysis tubing is 3500, again with redistilled water dialysis 12-24 hour, then dialysis tubing internal reaction liquid is spin-dried for the back and dissolves with the 5ml methylene dichloride afterwards, it is splashed into anhydrous diethyl ether, filter, vacuum-drying makes the FA-Pluronic-OH of a terminal modified FA.

2, FA-Pluronic-PLA's is synthetic: do macromole evocating agent and stannous octoate is a catalyzer with FA-Pluronic-OH, under the condition of anhydrous, anaerobic, (lactide LA) carries out ring-opening polymerization, finally obtains required multipolymer to cause the cyclic monomer rac-Lactide.Concrete synthetic method is: reaction flask vacuumizes-Tong argon gas deoxygenation dehumidifying after, under the argon gas condition, add FA-Pluronic-OH and rac-Lactide and stannous octoate, the amount of rac-Lactide is 50 – 90% of FA-Pluronic-OH weight, the amount of stannous octoate is the 0.1-0.15% of rac-Lactide weight, then reactant is heated to 140-160 ^oC stirs down, and reaction continues 6-8 hour; Then, reactant is sunk in the methyl alcohol, the adularescent material settles out, and filters; Use the methylene dichloride dissolve polymer again, and sink in the methyl alcohol, filter, drying finally obtains the FA-Pluronic-PLA multipolymer.

The present invention selects the Pluronic segmented copolymer, and following advantage is arranged:

1, Pluronic is a kind of business-like product, and the Pluronic product of a series of different molecular weights, PEO and PPO block ratio is arranged, and compares with PEO, and Pluronic has more selectivity;

2, Pluronic has good biocompatibility, and the Pluronic product that part PEO content is high is used by FDA (FDA) approval;

3, experimental results show that, the nanoparticle that the surface is formed by Pluronic, with specific surface is the nanoparticle of PEO, the easier cytolemma that passes, because it is similarly amphipathic that Pluronic has with cytolemma, and Pluronic has also found good application prospects at aspects such as gene therapy, cancer therapy.

The present invention selects PLA as hydrophobic segment, because it is the polyester of biocompatibility and biological degradability, is also used by drugs approved by FDA.It can be degraded into small molecules in human body, thereby is easy to be excreted, so it is widely used in biomedicine field.

The FA-Pluronic-PLA amphipathic nature block polymer that the present invention relates to is a polymkeric substance with new chemical structure.

Description of drawings

Fig. 1 is the chemical structural formula of FA-Pluronic-PLA multipolymer of the present invention;

Fig. 2 is the uv-spectrogram of FA and FA-F127-PLA multipolymer.

Embodiment

A kind of folic acid target polymer drug carrier that contains--the preparation method is as follows for folic acid-polyoxyethylene-poly-oxypropylene polyoxyethylene-poly(lactic acid) (FA-Pluronic-PLA), what we selected for use is that Pluronic F127 product synthesizes FA-F127-PLA, and the molecular formula of F127 is PEO ₁₀₀-PPO ₆₅-PEO ₁₀₀:

1, FA-F127-OH's is synthetic: earlier with 0.42 g, 0.95mmol FA and 0.18 g 1,3-dicyclohexylcarbodiimide (DCC) joins in the 35ml anhydrous dimethyl sulfoxide (DMSO), at room temperature stirred 12 hours, again with 10g, 0.79mmol Pluronic F127 and 0.097g 4-Dimethylamino pyridine (DMAP) add wherein, continue at room temperature to stir 24 hours.Afterwards with centrifugal 5 minutes of reactant.Get supernatant and dialysed 3 hours with DMSO, the molecular weight cut-off of dialysis tubing is 3500, afterwards again with redistilled water dialysis 24 hours.Dialysis tubing internal reaction liquid is spin-dried for the back with the dissolving of 5ml methylene dichloride again, it is splashed into anhydrous diethyl ether, filter, vacuum-drying makes the FA-F127-OH of 5g one terminal modified FA.

2, FA-F127-PLA's is synthetic: reaction flask by vacuumize-Tong argon gas deoxygenation dehumidifying after, under the argon gas condition, add 5 g FA-Pluronic-OH, rac-Lactide 2.5 g and stannous octoate 2.5 mg, reactant is heated to 150 ^OC, to stir down, reaction continues 6 hours; Reactant is sunk in the methyl alcohol, and the adularescent material settles out, and filters; And then use the methylene dichloride dissolve polymer, and sink in the methyl alcohol, to filter, drying finally obtains FA-F127-PLA multipolymer (carrier) 3.8g, and productive rate is 50.7%.

Structural characterization ¹H NMR (400 MHz, DMSO-d ⁶, ppm): 1.04-1.05 (m, CH ₃Of PPO block in Pluronic), 1.28-1.30 (m, CH ₂CH ₂CO of folate), 1.47-1.49 (m, CH ₃Of PLA block), 3.33-3.52 (m, OCH ₂CH ₂Of PEO block and OCH ₂CH of PPO block in Pluronic), 5.21-5.22 (m, CH of PLA block), 6.5-7.7 (d, benzene-H of folate).

The molecular weight of the FA-F127-PLA multipolymer for preparing and the segment content of PLA are calculated by the nuclear magnetic spectrogram of FA-Pluronic-PLA, the result is 27000 for the molecular weight (Mn) of FA-F127-PLA, PLA segmental content is 51.7 wt%, and the structural formula that also is final polymkeric substance is FA-PEO ₁₀₀-PPO ₆₅-PEO ₁₀₀-PLA ₁₉₄, K=194, m=100, n=65.The content of FA is obtained by the ultraviolet spectrophotometer test in the FA-F127-PLA multipolymer.Fig. 2 is the uv atlas of FA and FA-F127-PLA multipolymer, and by the ultraviolet absorption peak of the FA of 288nm place as can be known, FA successfully is connected on the end group of F127-PLA multipolymer.

Claims

1. one kind contains folic acid target polymer drug carrier, and it is characterized in that: its molecular structural formula is as follows

Figure 201110238131X100001DEST_PATH_IMAGE001

2. the folic acid target polymer drug carrier that contains as claimed in claim 1, its preparation method may further comprise the steps:

(1), FA-Pluronic-OH's is synthetic: with 0.38-0.49g folic acid and 0.18g 1, the 3-dicyclohexylcarbodiimide joins in the 35ml anhydrous dimethyl sulfoxide, at room temperature stirred 10-15 hour, again with 10g, 0.79mmol polyoxyethylene-poly-oxypropylene polyoxyethylene and 0.097g 4-Dimethylamino pyridine add wherein, at room temperature continue to stir 24-48 hour, reactant is centrifugal, getting supernatant dialysed 2-4 hour with anhydrous dimethyl sulfoxide, the molecular weight cut-off of dialysis tubing is 3500, afterwards again with redistilled water dialysis 12-24 hour, then dialysis tubing internal reaction liquid is spin-dried for the back with the dissolving of 5ml methylene dichloride, it is splashed into anhydrous diethyl ether, filter, vacuum-drying makes the FA-Pluronic-OH of a terminal modified FA;

(2), reaction flask vacuumize-Tong argon gas deoxygenation dehumidifying after, under the argon gas condition, add FA-Pluronic-OH, rac-Lactide and stannous octoate, the amount of rac-Lactide is 50 – 90% of FA-Pluronic-OH weight, the amount of stannous octoate is 0.1-0.15 % of rac-Lactide weight, then reactant is heated to 140-160 ^oC stirs down, and reaction continues 6-8 hour, and reactant is sunk in the methyl alcohol, and the adularescent material settles out, and filters, and uses the methylene dichloride dissolve polymer again, and sinks in the methyl alcohol, filters, and drying finally obtains the FA-Pluronic-PLA multipolymer.

3. as claimed in claim 1 contain folic acid target polymer drug carrier the treatment cancer drug carrier in application.