CN105214098A - Containing the pharmaceutical carrier and its preparation method and application of polyamino acid and polyphosphoric acid choline - Google Patents
Containing the pharmaceutical carrier and its preparation method and application of polyamino acid and polyphosphoric acid choline Download PDFInfo
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Abstract
The present invention relates to a kind of polymer nanocomposite pharmaceutical carrier and its preparation method and application, particularly relate to the pharmaceutical carrier and its preparation method and application containing polyamino acid and polyphosphoric acid choline, containing the pharmaceutical carrier of polyamino acid and polyphosphoric acid choline, the block copolymer forming nano-medicament carrier is di-block copolymer, comprises one section of polyamino acid and one section of polyphosphoric acid choline; The pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline provided by the invention, shown by cellular level and the research of animal level, compared with cancer therapy drug freely, these medicament-carried nano micelles are all greatly improved in drug effect, circulation time, safety, for a kind of clinical nano-medicament carrier leaned forward, also the timbering material that the nucleus organizational project that can be used for regenerative medicine is cultivated, macromolecule precursor medicament, the fields such as biomaterial surface modification.
Description
Technical field
The present invention relates to a kind of polymer nanocomposite pharmaceutical carrier and its preparation method and application, particularly relate to the pharmaceutical carrier and its preparation method and application containing polyamino acid and polyphosphoric acid choline.
Technical background
What use in malignant tumor chemotherapy has Cytotoxic cancer therapy drug, and while killing tumor cell on a large scale, also normal tissue and organ cause infringement, bring great toxic and side effects to patient.But, then effectively can reduce the toxic and side effects of free cancer therapy drug in chemotherapy after there is the macromolecule medicament carrier carrying medicament of certain nano-scale; On the other hand, the nano-medicament carrier granule with certain size utilizes enhancing infiltration retention effect (RPR effect) at tumor tissues place, carries cancer therapy drug in the passive target enrichment of tumor tissues place, is conducive to the drug effect improving chemotherapeutics.Particularly business-like
nano-medicament carrier is ratified to treat advanced ovarian cancer and breast carcinoma clinically by U.S. FDA, develops new polymer nanocomposite pharmaceutical carrier bring hope (clock Cortex et Radix Polygalae etc., Science Bulletin, 2015,60,1339-1351) to researcheres.Although
can the hydrophilic doxorubicin hydrochloride of payload reduce this cancer therapy drug toxic and side effects, but the liposome of this cavity structure can not the hydrophobic cancer therapy drug of load.And most of cancer therapy drugs such as paclitaxel, cisplatin, the camptothecine etc. of clinical practice are at present all hydrophobic drugs, can not by this carrier payload.In addition, due to the liposome essence of this carrier, critical micelle concentration (CriticalMicelleConcentration, CMC) value is higher, easily causes disintegration after greatly being diluted in blood, causes the leakage in advance of carrying medicament.
Summary of the invention
For above-mentioned technical problem, the invention provides a kind of pharmaceutical carrier, the polymer nano micelle built by block copolymer, as the stable carrier of hydrophobic anticancer drug, load can be used for the treatment of tumor with the polymer nano micelle carrier of conveying hydrophobicity chemotherapeutics.
Concrete technical scheme is:
Containing the pharmaceutical carrier of polyamino acid and polyphosphoric acid choline, the block copolymer forming nano-medicament carrier is di-block copolymer, comprises one section of polyamino acid and one section of polyphosphoric acid gallbladder alkali; The chemical constitution of di-block copolymer is:
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100; R group is: benzyl, hydrogen or sodium ion.
The preparation method of di-block copolymer, has two kinds, and first method is:
(1) synthesis of micromolecule initiator, its chemical constitution is such as formula shown in (2).
Its concrete synthetic method is: 1molN-tertbutyloxycarbonyl-1，2-diaminoethane and 3-4mol triethylamine are dissolved in tetrahydrofuran solvent, and under 0 DEG C and nitrogen protection, 2-3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, again be dissolved in dichloromethane, then add 10-20ml trifluoroacetic acid, stir 5 hours, be spin-dried for dichloromethane solvent, add 20-30ml sodium hydrate aqueous solution, make its pH more than 11, then use 100ml dichloromethane extraction 5 times, collect dichloromethane solution, after rotary evaporation of solvent, namely obtain formula (2) micromolecule initiator;
(2) synthesis of polyamino acid macromole evocating agent:
Utilize micromolecule initiator in step (1) at N, cause Pidolidone benzyl ester-ring inner-acid anhydride (BLG-NCA) monomer or aspartic acid benzyl ester-ring inner-acid anhydride (BLA-NCA) monomer ring-opening polymerisation in dinethylformamide solvent, under nitrogen protection and room temperature, react 72 hours; After reaction solution is concentrated in ether precipitating, obtain polyglutamic acid or the poly-aspartate macromole evocating agent of carbobenzoxy group protection after drying, chemical structural formula divides such as formula shown in (3):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2;
(3) synthesis of amphipathic diblock copolymer:
Described amphipathic diblock copolymer refers to that polyamino acid block is hydrophobic block, and polyphosphoric acid choline is hydrophilic block.
Step (2) gained formula (3) macromole evocating agent and polymerisable monomer 2-methacrylic acid phosphocholine ester are dissolved in the mixed solvent of dimethyl sulfoxide and methanol, wherein the volume ratio of methanol and dimethyl sulfoxide is 0.1-10, after deoxygenation, 1:2 in molar ratio, add catalyst cuprous bromide and 2,2-bipyridyl, carries out catalytic reaction 48h; Reaction solution concentrated silicagel column removing mantoquita, in ether, further precipitating purification obtains the di-block copolymer such as formula (4).
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100;
(4) synthesis of full hydrophilic di-block copolymer:
Described full hydrophilic di-block copolymer refers to that polyamino acid block and polyphosphoric acid choline block are hydrophilic block.
The di-block copolymer utilizing step (3) to prepare, mixes with trifluoroacetic acid or sodium hydroxide solution, stirs 48h and carries out the protection of de-benzyl ester; Reaction solution dialysis is purified, and then lyophilization, obtains formula (5) or the complete hydrophilic bi-block copolymer of formula (6):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100;
The second preparation method:
(1) synthesis of micromolecule initiator, its chemical constitution is such as formula shown in (7).
Its concrete synthetic method is: 1molN-tertbutyloxycarbonyl-1,2-ethylenediamine (CAS:57260-73-8) and 3-4mol triethylamine are dissolved in tetrahydrofuran solvent, under 0 DEG C and nitrogen protection, 2-3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, obtain the micromolecule initiator of formula (7) amido protecting:
(2) synthesis of polyphosphoric acid choline macromole evocating agent:
Step (1) gained formula (7) micromolecule initiator and polymerisable monomer 2-methacrylic acid phosphocholine ester are dissolved in methanol or ethanol jointly, after deoxygenation, 1:2 in molar ratio, adds catalyst cuprous bromide and 2,2-bipyridyl, carries out catalytic reaction 48h; Reaction solution concentrated silicagel column removing mantoquita, in ether, further precipitating purification obtains the polyphosphoric acid choline macromole evocating agent such as formula (8); Macromole evocating agent such as formula (8) is dissolved in methanol or ethanol again; mix with trifluoroacetic acid solution; react more than 1 hour under room temperature, after complete deaminate protection, it is amino polyphosphoric acid choline macromole evocating agent that dialysis lyophilizing obtains such as formula the one end of (9):
Wherein, polyphosphoric acid choline block polymerization degree m is 10-100;
(3) synthesis of amphipathic diblock copolymer:
To utilize in step (2) such as formula the polyphosphoric acid choline macromole evocating agent shown in (9) at N, Pidolidone benzyl ester-ring inner-acid anhydride (BLG-NCA) monomer or aspartic acid benzyl ester-ring inner-acid anhydride (BLA-NCA) monomer ring-opening polymerisation is caused in the mixed solvent of dinethylformamide and methanol, 72 hours are reacted under nitrogen protection and room temperature, wherein the volume ratio of methanol and DMF is 0.1-10; After reaction solution is concentrated in ether precipitating, obtain amphipathic diblock copolymer after drying, chemical structural formula divides as shown in first method Chinese style (4):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100;
(4) synthesis of full hydrophilic di-block copolymer:
The di-block copolymer utilizing step (3) to prepare, mixes with trifluoroacetic acid or sodium hydroxide solution, stirs 48h and carries out the protection of de-benzyl ester; Reaction solution dialysis is purified, and then lyophilization, obtains first method Chinese style (5) or the complete hydrophilic di-block copolymer shown in formula (6):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100.
Di-block copolymer based on polyamino acid and polyphosphoric acid choline prepared by the present invention, can separately in aqueous solution self assembly be the polymer micelle structure of nano-scale, then hydrophobic anticancer drug is added, hydrophobic anticancer drug physical property loads in its hydrophobic cores the most at last, is configured to nano-medicament carrier.Its concrete preparation method is as follows:
Amphipathic diblock copolymer shown in 20mg formula (4) is dissolved in 2ml mixed solvent, mixed solvent is that the chloroform of 1:1 and methanol form by volume, or be that the dichloromethane of 1:1 and methanol form by volume, in the ultra-pure water that fast drop stirs to 10ml or normal saline, stirred at ambient temperature 2-5 hour, decompression extracts organic solvent, obtains polymer nano micelle aqueous solution; Then hydrophobic anticancer drug is as amycin, paclitaxel, camptothecine etc., be 20:1-2:1 according to the mass ratio of di-block copolymer and cancer therapy drug, be dissolved in 1ml chloroform or dichloromethane, drop in above-mentioned nano-micelle aqueous solution, stirred at ambient temperature 2-5 hour, after organic solvent volatilizees naturally, cancer therapy drug is wrapped in the hydrophobic inner core with nano-micelle.
Di-block copolymer based on polyamino acid and polyphosphoric acid choline prepared by the present invention, can mix with the first physical property of hydrophobic anticancer drug, then in aqueous solution, self assembly is the nanometer polymer micelle of medicine carrying, is configured to nano-medicament carrier.Its concrete preparation method is as follows:
Amphipathic diblock copolymer shown in 20mg formula (4), with hydrophobic anticancer drug as amycin, paclitaxel, camptothecine etc., be 20:1-2:1 according to the mass ratio of di-block copolymer and cancer therapy drug, jointly be dissolved in the mixed solvent of 2ml chloroform/methanol or methylene chloride/methanol, the volume ratio of interim mixed solvent is 1, in the ultra-pure water that fast drop stirs to 10ml or normal saline, stirred at ambient temperature 2-5 hour, decompression extracts organic solvent, finally obtains drug-carrying polymer nano-micelle aqueous solution.
Di-block copolymer based on polyamino acid and polyphosphoric acid choline prepared by the present invention, its polyamino acid block can pass through with hydrophobic anticancer drug the hydrophobic inner core that chemical bond complexation builds nano-micelle, and polyphosphoric acid choline block builds the hydrophilic outer layer of nano-micelle, thus composition nano-medicament carrier.Its concrete preparation method is as follows: 20mg formula (5) or the full hydrophilic di-block copolymer shown in formula (6), first it is fully dissolved in ultra-pure water, then cancer therapy drug cisplatin is added, wherein the mol ratio of cisplatin and bi-block copolymer carboxyl is 2:1-1:5, stir more than 48 hours under room temperature condition, the hydrophobic inner core of nano-micelle is jointly configured to, the hydrophilic outer shell of polyphosphoric acid choline composition nano-micelle by the complexing of the carboxyl of cisplatin and polyamino acid.
The above hydrophobic anticancer drug is mainly amycin, paclitaxel, camptothecine and cisplatin, but is not limited to above hydrophobic anticancer drug.
The present invention has synthesized two class diblock copolymers, one class is the amphipathic diblock copolymer shown in (4) formula, its hydrophobic polyamino acids block forms the hydrophobic cores of nano-micelle, for load hydrophobic anticancer drug, hydrophilic phosphocholine forms the hydrophilic outer shell of nano-micelle, provides the biofacies same sex of nano-micelle excellence; Another kind of is the full hydrophilic di-block copolymer shown in formula (5) or formula (6), its hydrophilic polyamino acid block can chemical bond complexed cisplatin, the hydrophobic inner core of common composition nano-micelle, hydrophilic phosphocholine forms the hydrophilic outer shell of nano-micelle, provides the biofacies same sex of nano-micelle excellence.Two class nano-micelle carriers all can payload hydrophobicity chemotherapeutics.
Nano-micelle grain size before medicine carrying of the present invention after nano-micelle and medicine carrying is distributed between 10-200 nanometer.
The pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline provided by the invention, shown by cellular level and the research of animal level, compared with cancer therapy drug freely, these medicament-carried nano micelles are all greatly improved in drug effect, circulation time, safety, for a kind of clinical nano-medicament carrier leaned forward, also the timbering material that the nucleus organizational project that can be used for regenerative medicine is cultivated, macromolecule precursor medicament, the fields such as biomaterial surface modification.
Accompanying drawing explanation
The hydrogen spectrum nuclear-magnetism figure of Fig. 1 is one end to be the amino other end the be micromolecule initiator of acylbromide;
Fig. 2 is the hydrogen spectrum nuclear-magnetism figure of the polyamino acid macromole evocating agent of benzyl protection;
Fig. 3 is the hydrogen spectrum nuclear-magnetism figure of amphipathic diblock copolymer;
Fig. 4 is the hydrogen spectrum nuclear-magnetism figure of full hydrophilic di-block copolymer;
Fig. 5 is the grain size distribution carrying amycin polymer micelle;
Fig. 6 is the transmission electron microscope picture carrying amycin polymer micelle;
Fig. 7 is the polymer micelle grain size distribution that blank amphipathic nature block polymer builds;
Fig. 8 is the cytotoxicity figure of blank polymer micelle;
Fig. 9 is the cytotoxicity figure carrying amycin polymer micelle;
Figure 10 is the zoopery figure carrying amycin polymer micelle Tumor suppression.
Detailed description of the invention
The specific embodiment of the present invention is described in conjunction with the embodiments.
Embodiment 1
The preparation of the micromolecule initiator shown in formula (2):
1molN-tertbutyloxycarbonyl-1，2-diaminoethane and 3-4mol triethylamine are dissolved in tetrahydrofuran solvent, and under 0 DEG C and nitrogen protection, 2-3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, again be dissolved in dichloromethane, then add 10-20ml trifluoroacetic acid, stir 5 hours, be spin-dried for dichloromethane solvent, add 20-30ml sodium hydrate aqueous solution, make its pH more than 11, then use 100ml dichloromethane extraction 5 times, collect dichloromethane solution, after rotary evaporation of solvent, namely obtain the micromolecule initiator shown in (2).The hydrogen spectrum nuclear-magnetism figure of this micromolecule initiator is shown in Fig. 1.
The preparation of the polyamino acid macromole evocating agent shown in formula (3):
Get the micromolecule initiator of preparation in 0.1mol embodiment 1; the anhydrous N of 70ml is dissolved in 3mol aspartic acid benzyl ester-ring inner-acid anhydride (BLA-NCA) monomer or 5mol benzyl glutamate-ring inner-acid anhydride (BLA-NCA); in dinethylformamide solvent; under nitrogen protection and normal temperature condition; stirring reaction 72h; reaction terminates rear rotary evaporation concentrated reaction solution; precipitating 3 times in ether, obtains the polyamino acid macromole evocating agent shown in chemical formula (3) after drying.The hydrogen spectrum nuclear-magnetism figure of this macromole evocating agent is shown in Fig. 2.
The preparation of the amphipathic diblock copolymer shown in formula (4):
Macromole evocating agent 1mmol in modus ponens (3), jointly be dissolved in the mixed solvent of 50ml dimethyl sulfoxide and methanol with 30 or 50mmol2-metering system phosphocholine ester monomer, wherein the volume ratio of methanol and dimethyl sulfoxide is 0.1-10, after abundant deoxygenation, add 1mmol cuprous bromide and 2mmol2,2-bipyridyl, atom transfer radical polymerization 48h under normal temperature condition; Cross silicagel column removing mantoquita after reaction solution is concentrated, then precipitating 2 times in ice ether, obtains the amphipathic diblock copolymer shown in formula (4) after product drying.The nuclear magnetic spectrogram of this amphipathic diblock copolymer is shown in Fig. 3.
Embodiment 2
The preparation of the full hydrophilic di-block copolymer shown in formula (6):
The amphipathic diblock copolymer 200mg of modus ponens (4); be dissolved in 20ml ultra-pure water according to 5 times of weighing sodium hydroxide of polyamino acid repetitive molal quantity in polymer; and add the amphipathic diblock copolymer of above-mentioned amount; stir 72 hours under normal temperature condition; slough benzyl protection; after reaction solution dialysis is purified, lyophilization, obtains the full hydrophilic di-block copolymer shown in formula (6).Its hydrogen spectrum nuclear-magnetism figure is shown in Fig. 4.
Embodiment 3
The preparation of micromolecule initiator shown in formula (7):
1molN-tertbutyloxycarbonyl-1，2-diaminoethane 4mol triethylamine is dissolved in the tetrahydrofuran solvent after dewatering, and under 0 DEG C and nitrogen protection, 3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, obtain the micromolecule initiator of amido protecting.
The preparation of the polyphosphoric acid choline macromole evocating agent shown in formula (9):
Get the micromolecule initiator of 1mmol formula (7), jointly be dissolved in 50ml methanol with 30 or 50mmol2-metering system phosphocholine ester monomer, after abundant deoxygenation, add 1mmol cuprous bromide and 2mmol2,2-bipyridyl, atom transfer radical polymerization 48h under normal temperature condition; Cross silicagel column removing mantoquita after reaction solution is concentrated, then precipitating 2 times in ice ether, obtains the polyphosphoric acid choline polymer of the band amido protecting shown in formula (8) after product drying; The polyphosphoric acid choline polymer getting this band amido protecting of 100mg is dissolved in 10ml methanol and mixes with 10ml trifluoroacetic acid solution; react 2 hours under room temperature; after complete deaminate protection, it is amino polyphosphoric acid choline macromole evocating agent that dialysis lyophilizing obtains such as formula the one end of (9).
The preparation of the amphipathic diblock copolymer shown in formula (4):
Formula (9) one end prepared of getting previous step is amino polyphosphoric acid choline macromole evocating agent 0.1mmol, the anhydrous N of 70ml is dissolved in 3mmol aspartic acid benzyl ester-ring inner-acid anhydride (BLA-NCA) monomer or 5mmol benzyl glutamate-ring inner-acid anhydride (BLA-NCA), in the mixed solvent of dinethylformamide and methanol, wherein methanol and N, the volume ratio of dinethylformamide is 0.1-10, under nitrogen protection and normal temperature condition, stirring reaction 72h, reaction terminates rear rotary evaporation concentrated reaction solution, precipitating 3 times in ether, the amphipathic diblock copolymer shown in formula (4) is obtained after drying.
Embodiment 4
The preparation of the full hydrophilic di-block copolymer shown in formula (6):
The amphipathic diblock copolymer 200mg of preparation in Example 3; be dissolved in 20ml ultra-pure water according to 5 times of weighing sodium hydroxide of polyamino acid repetitive molal quantity in polymer; and add the amphipathic diblock copolymer of above-mentioned amount; stir 72 hours under normal temperature condition; slough benzyl protection; after reaction solution dialysis is purified, lyophilization, obtains the full hydrophilic di-block copolymer shown in formula (6).
Embodiment 5
Amphipathic diblock copolymer shown in formula (4) and hydrophobic anticancer drug amycin Hybrid assembling medicament-carried nano micelle:
Get in the chloroform and methanol mixed solvent that the amphipathic diblock copolymer shown in 20mg formula (4) and the hydrophobic amycin of 8mg be dissolved in 1ml volume ratio 1:1 jointly, and in the ultra-pure water that stirs to 10ml of fast injection, after decompression removing organic solvent, obtain the carrier micelle of load amycin.Particle diameter and the transmission electron microscope picture of carrier micelle are shown in Fig. 5 and Fig. 6 respectively.
Load amycin micelle is applied the toxicity of breast cancer cell: prepare according to embodiment 5 carrier micelle that a series of gradient medicine makes, and the free doxorubicin hydrochloride preparing identical gradient drug concentration in contrast, with breast cancer cell Dual culture 72 hours, mtt assay is utilized to measure the activity of cell, result shows that carrier micelle effectively can kill cancerous cell, the results are shown in Figure 9.
In zoopery, load amycin micelle is applied the suppression of tumor: get the nude mice that 15 are implanted with breast cancer tumour model, be divided into three groups, often organizes 5.Every three days intravenous injection sodium chloride 200 microlitres, free amycin (DOX) 5.0mg/kg, the medicament-carried nano micelle in embodiment 5, equivalent DOX5.0mg/kg.Measured with the volume of slide calliper rule to tumor every three days.The computing formula of gross tumor volume is as follows: volume (mm
3the long * of)=0.58* is wide
2.Experimental result is shown in Figure 10.Result shows to compare with sodium chloride group with injection free drug, and tumor obtains suppression to a certain extent.
Embodiment 6
Amphipathic diblock copolymer shown in formula (4) and hydrophobic anticancer drug paclitaxel Hybrid assembling medicament-carried nano micelle:
Get in the chloroform and methanol mixed solvent that the amphipathic diblock copolymer shown in 20mg formula (4) and 4mg paclitaxel be dissolved in 1ml volume ratio 1:1 jointly, and in the ultra-pure water that stirs to 10ml of fast injection, after decompression removing organic solvent, obtain paclitaxel loaded carrier micelle.
Embodiment 7
Medicament-carried nano micelle is assembled in full hydrophilic di-block copolymer shown in formula (6) and the complexation of hydrophobic anticancer drug cisplatin:
Getting the full hydrophilic di-block copolymer shown in 20mg formula (6) is dissolved in 10ml ultra-pure water, take the equimolar cisplatin of polyamino acid repetitive in full hydrophilic di-block copolymer, join in above-mentioned aqueous solution, complex reaction 72 hours under room temperature, utilize the non-complexed cisplatin of the centrifugal removing of ultrafilter membrane, obtain the carrier micelle of load cisplatin.
Embodiment 8
Amphipathic diblock copolymer shown in formula (4) prepares blank polymeric micelle:
Get in the chloroform and methanol mixed solvent that the amphipathic diblock copolymer shown in 20mg formula (4) is dissolved in 1ml volume ratio 1:1, and in the ultra-pure water that stirs to 10ml of fast injection, after decompression removing organic solvent, obtain the polymer micelle that non-medicine carrying is namely blank.The grain size distribution of blank polymer micelle is shown in Fig. 7.
The cytotoxicity of blank polymer micelle: prepare the blank polymer micelle of a series of gradient concentration and breast cancer cell Dual culture after 24 hours according to embodiment 8, mtt assay is utilized to measure the activity of cell, result shows that blank polymer micelle has good biocompatibility, the results are shown in Figure 8.
Claims (8)
1. contain the pharmaceutical carrier of polyamino acid and polyphosphoric acid choline, it is characterized in that, the block copolymer forming nano-medicament carrier is di-block copolymer, comprises one section of polyamino acid and one section of polyphosphoric acid gallbladder alkali; The chemical constitution of di-block copolymer is:
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100; R group is benzyl, hydrogen or sodium ion.
2. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 1, is characterized in that its preparation method comprises the following steps:
(1) synthesis of micromolecule initiator; 1molN-tertbutyloxycarbonyl-1，2-diaminoethane and 3-4mol triethylamine are dissolved in tetrahydrofuran solvent, and under 0 DEG C and nitrogen protection, 2-3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, again be dissolved in dichloromethane, then add 10-20ml trifluoroacetic acid, stir 5 hours, be spin-dried for dichloromethane solvent, add 20-30ml sodium hydrate aqueous solution, make its pH more than 11, then use 100ml dichloromethane extraction 5 times, collect dichloromethane solution, after rotary evaporation of solvent, namely obtain the micromolecule initiator of formula (2);
(2) utilize the micromolecule initiator of step (1) gained at N, cause Pidolidone benzyl ester-ring inner-acid anhydride monomer or aspartic acid benzyl ester-ring inner-acid anhydride monomer ring-opening polymerisation in dinethylformamide solvent, under nitrogen protection and room temperature, react 72 hours; After reaction solution is concentrated in ether precipitating, obtain the polyglutamic acid of carbobenzoxy group protection or the macromole evocating agent of poly-aspartate after drying, macromole evocating agent chemical structural formula is formula (3):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2;
(3) step (2) gained macromole evocating agent and polymerisable monomer 2-methacrylic acid phosphocholine ester are dissolved in the mixed solvent of dimethyl sulfoxide and methanol, wherein the volume ratio of methanol and dimethyl sulfoxide is 0.1-10, after deoxygenation, 1:2 in molar ratio, add catalyst cuprous bromide and 2,2-bipyridyl, carries out catalytic reaction 48h; Reaction solution concentrated silicagel column removing mantoquita, in ether, further precipitating purification obtains the di-block copolymer of formula (4);
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100;
(4) di-block copolymer utilizing step (3) to prepare, mixes with trifluoroacetic acid or sodium hydroxide solution, stirs 48h and carries out the protection of de-benzyl ester; Reaction solution dialysis is purified, and then lyophilization, obtains complete hydrophilic bi-block copolymer (5) or (6):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100.
3. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 1, is characterized in that its preparation method comprises the following steps:
(1) synthesis of micromolecule initiator; 1molN-tertbutyloxycarbonyl-1，2-diaminoethane and 3-4mol triethylamine are dissolved in tetrahydrofuran solvent, and under 0 DEG C and nitrogen protection, 2-3mol2-bromo isobutyl acylbromide dropwise adds, and reacts 24 hours; Cross and filter triethylamine salt, by filtrate rotary evaporation to white solid state, obtain the micromolecule initiator of amido protecting, structural formula is formula (7):
(2) step (1) gained formula (7) micromolecule initiator and polymerisable monomer 2-methacrylic acid phosphocholine ester are dissolved in methanol or ethanol jointly, after deoxygenation, 1:2 in molar ratio, add catalyst cuprous bromide and 2,2-bipyridyl, carries out catalytic reaction 48h; Reaction solution concentrated silicagel column removing mantoquita, in ether, further precipitating purification obtains the polyphosphoric acid choline macromole evocating agent of formula (8); The macromole evocating agent of formula (8) is dissolved in methanol or ethanol again; mix with trifluoroacetic acid solution; react more than 1 hour under room temperature, after complete deaminate protection, one end that dialysis lyophilizing obtains formula (9) is amino polyphosphoric acid choline macromole evocating agent:
Wherein, polyphosphoric acid choline block polymerization degree m is 10-100;
(3) utilize the polyphosphoric acid choline macromole evocating agent shown in step (2) Chinese style (9) at N, Pidolidone benzyl ester-ring inner-acid anhydride monomer or aspartic acid benzyl ester-ring inner-acid anhydride monomer ring-opening polymerisation is caused in the mixed solvent of dinethylformamide and methanol, 72 hours are reacted under nitrogen protection and room temperature, wherein the volume ratio of methanol and DMF is 0.1-10; After reaction solution is concentrated in ether precipitating, obtain bi-block copolymer after drying, chemical structural formula is formula (4):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100;
(4) di-block copolymer utilizing step (3) to prepare, mixes with trifluoroacetic acid or sodium hydroxide solution, stirs 48h and carries out the protection of de-benzyl ester; Reaction solution dialysis is purified, and then lyophilization, obtains the complete hydrophilic bi-block copolymer of formula (5) or formula (6):
Wherein, the polymerization degree n of polyamino acid block is 10-100, and its pendent methylene number k is 1 or 2; Polyphosphoric acid choline block polymerization degree m is 10-100.
4. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to any one of Claim 1-3, is characterized in that: application and the polymer micelle building nano-scale, becomes the carrier of load and conveying cancer therapy drug and realizes the controllable release of medicine.
5. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 4, is characterized in that: described di-block copolymer forms nano-micelle by self assembly in aqueous solution, is then carried on wherein by cancer therapy drug; Comprise the following steps:
Utilize the di-block copolymer of formula (4) and hydrophobic anticancer drug to be jointly dissolved in mixed solvent, described mixed solvent is that methanol and chloroform mix according to volume ratio 1:1, or methanol mixes according to volume ratio 1:1 with dichloromethane; The mass ratio of di-block copolymer and cancer therapy drug is 20:1-2:1, the mixed solution of bi-block copolymer and cancer therapy drug instills to ultra-pure water or in normal saline under agitation, stirred at ambient temperature 2-5 hour, decompression extracts organic solvent, finally obtains the nano-medicament carrier of load hydrophobic anticancer drug.
6. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 5, is characterized in that: described cancer therapy drug is hydrophobicity amycin, paclitaxel or camptothecine.
7. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 4, is characterized in that: described di-block copolymer and the mixture of cancer therapy drug form medicament-carried nano micelle by self assembly in aqueous solution; Comprise the following steps:
Utilize the full hydrophilic di-block copolymer of formula (5) or formula (6), fully be dissolved in ultra-pure water, then cancer therapy drug cisplatin is added, wherein the mol ratio of cisplatin and bi-block copolymer carboxyl is 2:1-1:5, stir more than 48 hours under room temperature condition, final reaction solution removes the cisplatin medicine of non-load under utilizing ultrafilter membrane centrifugal condition, finally obtain the polymer nano micelle of load cisplatin.
8. the pharmaceutical carrier containing polyamino acid and polyphosphoric acid choline according to claim 4, is characterized in that: described nano-micelle is of a size of 10-200nm.
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