CN102504245A - Amphipathic amino acid block copolymer and preparation method and application thereof - Google Patents
Amphipathic amino acid block copolymer and preparation method and application thereof Download PDFInfo
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- CN102504245A CN102504245A CN 201110340246 CN201110340246A CN102504245A CN 102504245 A CN102504245 A CN 102504245A CN 201110340246 CN201110340246 CN 201110340246 CN 201110340246 A CN201110340246 A CN 201110340246A CN 102504245 A CN102504245 A CN 102504245A
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- amino acid
- amphipathic
- segmented copolymer
- amphipathic amino
- acid
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- 238000002360 preparation method Methods 0.000 title claims abstract description 54
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- 238000000034 method Methods 0.000 claims abstract description 21
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical group N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims abstract description 18
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- 229940024606 amino acid Drugs 0.000 claims description 113
- 229920001577 copolymer Polymers 0.000 claims description 73
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 51
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- 238000003756 stirring Methods 0.000 claims description 39
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Abstract
The invention relates to amphipathic amino acid block copolymer and a preparation method and application thereof. The invention discloses the amphipathic amino acid block copolymer with a general formula (I) and salt thereof, wherein m is a positive integer in the range of 1 to 200; n is a positive integer in the range of 1 to 150; X is active terminal ammonia; and Y is alkali metal. The block copolymer can be prepared by performing reaction of anhydride of one amino acid and the active terminal ammonia, then performing reaction of the obtained product and anhydride of another amino acid and removing benzyl by adopting alkali hydroxide. Micelle prepared from the polymer has low critical micelle concentration, is not influenced by pH, has high stability and high drug-loading rate, can be used as a carrier of an organic medicament and waterinsoluble or insoluble and amphipathic medicaments, and is used for endovascular or intramuscular injection, oral administration and tract or external application drug administration. The preparation method is simple. The process is mature. The amphipathic amino acid block copolymer is suitable for large-scale continuous production.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, relate to the amphipathic amino acid segmented copolymer of a kind of biodegradability, the invention still further relates to the preparation method and the application thereof of this carrier as pharmaceutical carrier.
Background technology
The research of modern medicines transfer system be unable to do without the use of type material; Pharmaceutical polymers particularly, chitosan, mucinase, gelatin, starch, sodium-alginate, BSA, Mierocrystalline cellulose, POLYACTIC ACID, Z 150PH, acrylic acid or the like, polyester, polyethers etc. have shown special advantages in pharmaceutics.But biocompatibility, biological degradability and security have restricted the use of pharmaceutical polymers, and the macromolecular material of developing safe new function belongs to the hot fields of pharmaceutics research.Amino acid is the essentially consist unit of biological function macro-molecular protein, is the base substance that constitutes body nutrition desired protein.Select for use one or more amino acid synthetic polymers to be degraded to amino acid under the effect of enzyme in vivo, good biocompatibility, safety non-toxic, the research of polyamino acid has received widely and having paid close attention to.
Polyamino acid mainly concentrates on polyamino acid-drug conjugates, polyamino acid complex carrier, the several aspects of amino acid copolymer carrier as the research of pharmaceutical carrier at present.Polyamino acid and medicine form conjugate through chemical bond, and chemical bond rupture discharges medicine under the effect of sour environment and enzyme in vivo, reaches the effect of slowly-releasing, target, and can reduce the toxicity of medicine.Polyamino acid and other macromolecular material form complex carrier to overcome the not enough of single-material and to realize new function.Amino acid copolymer also can form amphipathic nature material as pharmaceutical carrier, to improve the medicine dissolution performance, extension body internal recycle time and realization target purpose.
1. polyamino acid-drug conjugates
Polyamino acid contains active amino and carboxyl terminal; Can combine with medicine through covalent linkage, form conjugate, chemical bond rupture discharges medicine under the effect of specific in vivo sour environment of this conjugate and enzyme; Reach the effect of slowly-releasing, target; Owing to the provide protection of polyamino acid, improve stability of drug simultaneously, and can reduce the toxicity of medicine.Polyamino acid-drug conjugates research originates from the early seventies in last century; The U.S. still its kim of university leader's seminar has just successfully synthesized the polyglutamic acid material; Use aminopropanol to be side chain radical; With the covalent attachment mode contraceptive steroid Norethisterone is keyed in material, and carried out external and the rat vivo releasing test, reach 300d its time of releasing.Experimental result shows that polyglutamic acid degradable after degraded is that monomer is not detained in vivo.
CN1321696A, CN101209350A, CN101707869A, US6262107, US6515017, US6730699, US6884817 is disclosed all to be to adopt polyamino acid and medicine to prepare the method for conjugate through the chemical bond coupling.Because such carrier is through medicine and the coupling of polyamino acid covalent linkage; All need design proper reaction conditions to each medicine; And accomplish preparation, the purifying of polyamino acid-drug conjugates, increased the difficulty of implementing, drug loss big, improved cost.
2. polyamino acid complex carrier
Polyamino acid combines to form complex carrier through chemical bond or physical action with macromolecular material commonly used, can realize transfection efficiency, target, the purpose of extension body internal recycle time of slowly-releasing, reduction toxicity, raising bio-pharmaceutical.
The disclosed polyamino acid triblock copolymer of CN101619132B can be used for preparing gel.The pharmaceutical prepn of this method preparation is inappropriate for intravenous injection.
The dendritic polyamino acid of the disclosed preparation of US20030232968A1, complicated process of preparation, and dendritic structure also produces immunoreation than linear structure.
US20030147958A1 is disclosed with polyamino acid and hydrophilic polymer such as polyoxyethylene glycol (PEG) coupling, improves transfection efficiency, reduces toxicity.Other has the lot of documents report to adopt PEG and polyamino acid coupling as pharmaceutical carrier.As: (Prompruk K such as Prompruk K; Govender T, Zhang S, et al.Synthesis of a novel PEG-block-poly (aspartic acid-statphenylalanine) copolymer shows potential for formation of a micellar drug carrier [J] .International Journal of Pharmaceutics; 2005; 297 (1-2): 242～253) with PEG be hydrophilic section, aspartic acid and phenylalanine(Phe) (Phenylalanine) stochastic sequence synthetic polymkeric substance forms segmented copolymer as hydrophobic section; Aspartic acid can be through ionization bag medicine carrying thing; And the hydrophobicity of phenylalanine(Phe) or aromatic ring interaction help the micella nucleation, slow down micella to dissociate, and delay the release of medicine; (Kim SH such as Kim SH; Jeong JH, Joe CO, et al.Folate receptor mediated intracellular protein delivery using PLL-PEG-FOL conjugate [J] .Journal of Controlled Release; 2005; 103 (3): 625～634.) preparation polylysine PEG amphipathic nature block polymer, and, form the PLL-PEG-FOL polymkeric substance of positively charged with modified with folic acid PEG; (Shu SJ such as Shu SJ; Zhang XG.; Teng DY; Et al.Polyelectrolyte nanoparticles based on water-soluble chitosan-poly (L-aspartic acid)-polyethylene glycol for controlled protein release [J] .Carbohydr-ate Research, 2009,344 (10): 1197～1204.) preparation water-soluble chitosan-SAP 73-PEG (WSC-Pasp-PEG) mixture.
Adopt polyamino acid and PEG to prepare complex carrier and can reach effects such as target, raising drug loading, slowly-releasing, the interior long circulation of body; But the process that PEG modifies has increased the cost of preparation and has produced impurity; PEG modifies the back and shows the uriniferous tubules cavity after the medication repeatedly at animal model; In addition, also can produce antibody at animal and human's body behind the Using P EG.
Adopt safer wetting ability polyamino acid to substitute PEG and also can reach long circulation work.(Romberg B, Metselaar JM, Baranyi L such as Romberg B; Et al.Poly (amino acid) s:Promising enzymeatically degradable stealth coatings for liposomes [J] .International Journal of Pharmaceutics; 2007,331 (2): 186～189.) use 5% (mol ratio) SAP 73 (PHEA, molecular weight 3000Da), polyglutamic acid (PHEG respectively; Molecular weight 4000Da) modified liposome; And compare its transformation period, do not wrap up liposome transformation period 9.7h, the liposome transformation period of wrapping up with SAP 73, polyglutamic acid is respectively 14.4h and 11.9h.Adopt wetting ability polyamino acid modified liposome can reach long Circulation, but the process of modifying has increased the complicacy of technology, stability of formulation also is difficult to reach requirement.
3. amino acid copolymer carrier
Adopt one or more amino acid synthesizing amino acid copolymers as pharmaceutical carrier and since meta-bolites be amino acid to human non-toxic, have higher security than other macromolecular material; Origin of amino acid is abundant, low price, and forming the human amino acid has 20 kinds; Can select as required; Satisfy the needs of different pharmaceutical carrier, have a lot of reactive groups (as: amino, carboxyl) on the polyamino acid molecular chain, medicine or actives mass-energy are convenient to be combined with it.Such carrier is compared the preparation of polyamino acid-drug conjugates simply, does not need the complicated purification process, with polyamino acid complex carrier simplified in comparison technology, preparation stabilization, the medicine drug loading is high, security is higher.Therefore amino acid copolymer also more and more receives publicity as pharmaceutical carrier.
(what expects Han Yadong etc. for Han Yadong, Xia Jialiang; Deng. L-glutamic acid supports the preparation and the evaluation [J] of oral insulin microballoon. SCI; 2009,30 (12): be solid support material 2521～2524.), adopt anhydrous emulsion legal system to be equipped with the oral insulin microballoon with the L-glutamic acid; Microsphere diameter is at 5～20 μ m, and the medicine carrying massfraction is 5%～9%.But this method drug loading is low, and the particle diameter of preparation is not suitable for intravenously administrable at 5～20 μ m.
(Akagi T such as Akagi T; Kaneko T; Kida T; Et al.Preparation and characterization of biodegradable nanoparticles based on poly (g-glutamic acid) with l-phenylalanine as a protein carrier [J] .Journal of Controlled Release; 2005,108 (2-3): 226～236.) (γ-PGA) and the graft copolymerization of L-phenylalanine ethyl ester, this carrier is with ovalbumin parcel preparation nanoparticle with gamma-polyglutamic acid-.The encapsulation rate of this method medicine is below 50%, and encapsulation rate is low, is prone to after the lyophilize assemble.
(Kima MS such as Kima MS; Dayananda K, Choi EK, et al.Synthesis and characterization of poly (L-Glutamic acid)-block-poly-(L-phenylalanine) [J] .Polymer; 2009; 50 (10): 2252～2257) supply polymers with L-glutamic acid, gamma-benzyl ester and the synthetic block of L-phenylalanine(Phe), after sloughing the benzyl ester, form amphipathic copolymer vesicle, this carrier forms the vesicle of different-grain diameter under condition of different pH.This carrier does not have the application report as pharmaceutical carrier; The vesicle particle diameter that under condition of different pH, forms of this carrier is different in addition, though can utilize this attributes design pH sensitive drug carrier, has explained that also there is the pH unstable in this carrier; Because most drug is only just stable in a certain pH scope; This just require preparation preparation, to control pH when storing, bring into play the sharpest edges of this carrier surely but under this pH condition, differ, in addition; The fluctuation of pH also can take place in formulation preparation, storage process, and pH sensitive drug carrier can produce stability problem.
Thus it is clear that, adopt polyamino acid to have defectives such as technology is loaded down with trivial details, complex structure is prone to produce immunogenicity, and the nano-carrier bag medicine carrying thing of formation is limited, and preparation stability is not good at present as pharmaceutical carrier.
The amino acid of forming human body protein has 20 kinds, and the inventor finds that if adopt hydrophilic L-aspartic acid and hydrophobicity L-phenylalanine(Phe) to prepare amphipathic amino acid segmented copolymer, self-assembly forms nano-micelle in the aqueous solution, and the preparation method is simple; This amphipathic amino acid segmented copolymer has pH stability; Under the hydrophobic grouping effect, can significantly improve the drug loading of medicine in addition, steady time is significant prolongation also.The present invention adopts L-aspartic acid and L-phenylalanine(Phe) to prepare amphipathic amino acid segmented copolymer, and self-assembled nano micelle is not seen any document and patent report.
Summary of the invention
The object of the invention to the problems referred to above provide a kind of with L-aspartic acid-beta benzyl ester ring inner-acid anhydride and L-phenylalanine(Phe) ring inner-acid anhydride as starting raw material, through polyreaction obtain that micelle-forming concentration is low, drug loading is high, good stability, the medicament solubilization carrier that not influenced by pH.
Another object of the present invention provides the preparation method of above-mentioned carrier.
A further object of the invention provides the application of above-mentioned carrier in pharmacy.
For achieving the above object, the present invention provides a kind of newtype drug solubilization carrier, and this carrier adopts SAP 73 as hydrophilic chain, and polyphenylalanine makes it have amphipathic characteristic as hydrophobic chain, but self-assembly is a nano-micelle in water medium.
The preparation method of described medicament solubilization carrier comprises the following steps: L-aspartic acid-beta benzyl ester ring inner-acid anhydride is added active end group ammonia with N dissolving back, and room temperature condition stirs 12～72h down; Add anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product, collecting precipitation, dissolve with N dry back; Add L-phenylalanine(Phe) ring inner-acid anhydride again; Room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product, collecting precipitation; Drying promptly gets aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer; Or earlier L-phenylalanine(Phe) ring inner-acid anhydride being added active end group ammonia with N dissolving back, room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product; Collecting precipitation, dry back adds L-aspartic acid-beta benzyl ester ring inner-acid anhydride again with the N dissolving; Room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product, collecting precipitation; Drying promptly gets aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer; Take by weighing aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer, adding concentration is the NaOH solution of 0.5～2M, stirring at room 1～4h, and solution promptly gets amphipathic amino acid segmented copolymer through the dialysis tubing after drying of dialysing.
Described medicament solubilization carrier, wherein the L-aspartic acid is 1: 5～5: 1 with L-phenylalanine(Phe) molecule number ratio.
Described medicament solubilization carrier, its molecular weight are 1KDa～50KDa.
The application of described medicament solubilization carrier is characterized in that being used in the blood vessel or the pharmaceutical active of intramuscular injection or oral or cavity or external application or the carrier of pharmacologically active molecule.Wherein pharmaceutical active or pharmacologically active molecule are selected from: retinoid, taxanes, camptothecin, ciclosporin class, flavonoid 、 Xiao ? arbitrary material or derivatives thereof in bases, dihydropyridines, vinca, podophillotoxines, anthraquinone series antineoplastic medicament, steroidal class or nonsteroidal anti-inflammatory drug, microbiotic, cardiovascular agent, antiviral, antifungal drug, the immunomodulator.
The preparation method of this medicament solubilization carrier may further comprise the steps: amphipathic amino acid segmented copolymer and water obtain the nano-micelle of amphipathic amino acid segmented copolymer by weight the ratio dissolving that is 1～50: 1000; With the indissoluble of therapeutic dose or the organic drug that is slightly soluble in water with after the acceptable solvent dissolving pharmaceutically; After said amphipathic amino acid block copolymer nano micella mixes; Handle through ultrasonic or high pressure homogenization; Solvent is removed organic solvent and small molecules with dialysis tubing dialysis or ultrafiltration process or post partition method, and drying makes the nano-micelle that particle diameter is 10～1000nm.
Concrete scheme is following:
One, the compound method of amphipathic amino acid segmented copolymer is following:
1. the synthetic L-aspartic acid-beta benzyl ester ring inner-acid anhydride of SAP 73-β benzyl ester or polyphenylalanine dissolves with N, adds active end group ammonia, stirring at room 12～72h; Add absolute ethyl alcohol or anhydrous diethyl ether; Collecting precipitation, drying promptly gets SAP 73-β benzyl ester;
Or L-phenylalanine(Phe) ring inner-acid anhydride dissolves with N, adds active end group ammonia, and stirring at room 12～72h adds absolute ethyl alcohol or anhydrous diethyl ether, collecting precipitation, and drying promptly gets polyphenylalanine.
2. synthesizing of amphipathic aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer
SAP 73-β benzyl ester adds L-phenylalanine(Phe) ring inner-acid anhydride stirring at room 12～72h with the N dissolving, adds absolute ethyl alcohol or anhydrous diethyl ether, and collecting precipitation is dry, promptly gets amphipathic aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer;
Or polyphenylalanine N dissolving adding L-aspartic acid-beta benzyl ester ring inner-acid anhydride stirring at room 12～72h, add absolute ethyl alcohol or anhydrous diethyl ether, collecting precipitation, drying promptly gets amphipathic aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer.
3. synthesizing of amphipathic amino acid segmented copolymer
Take by weighing amphipathic aspartic acid-beta benzyl ester-phenylalanine(Phe) segmented copolymer, add 0.5～2M NaOH solution stirring at room, 1～4h, add the dialysis tubing dialysis, drying promptly gets amphipathic amino acid segmented copolymer.
Two, amphipathic amino acid block copolymer micelle preparation method
In the ratio of dissolving the amphipathic amino acid segmented copolymer of 1～20mg in every 1mL water; The amphipathic amino acid segmented copolymer that makes is soluble in water; (frequency of supersound process and treatment time all are that those of ordinary skills are known through supersound process; Down together), being prepared into particle diameter is the amphipathic amino acid block copolymer micelle of 10～1000nm.
Three, be carrier with amphipathic amino acid segmented copolymer, preparation contains the compsn of insoluble drug
After amphipathic amino acid segmented copolymer is water-soluble; Insoluble drug is dissolved with appropriate solvent (dissolving of pharmaceutically using that can dissolve this medicine); With amphipathic amino acid block copolymer solution through supersound process, make the polymer micelle of particle diameter 10～1000nm.
Four, adopt amphipathic amino acid segmented copolymer as the pharmaceutical carrier pharmaceutical compositions, insoluble drug is had solublization.
Can use this amphipathic amino acid segmented copolymer to comprise as the insoluble drug of carrier: all-trans-retinoic acid, taxol, Zorubicin, ciclosporin, NSC 94600, NSC 107124, nimodipine, Vumon, etoposide, Breviscarpine, bilobalide, SLB, daunorubicin, MTC, methotrexate, indomethacin, Ibuprofen BP/EP, Naproxen Base, but be not limited to listed medicine.
Beneficial effect of the present invention:
One, (Critical micelle concentrations, CMC): the CMC of amphipathic amino acid segmented copolymer all in 10～200mg/L scope, indicates that it has fabulous dilution stability to lower micelle-forming concentration among the embodiment 2～9.
Two, do not receive the influence of pH: adopt blank micella particle diameter in pH3.6, Ph4.0, pH4.6, pH5.0, pH5.6, pH6.0, pH6.5, pH7.0, pH7.4, pH8.0 solution of amphipathic amino acid segmented copolymer preparation significantly not change, do not have carrier micelle particle diameter, drug loading, steady time noticeable change.
Three, organic drug, water-insoluble or insoluble drug and amphipathic medicine had load preferably; For example; To the load of all-trans-retinoic acid up to 31.1% (w/w), to the load of taxol up to 28.4% (w/w), to the load of Zorubicin up to 27.5% (w/w); To the load of ciclosporin A up to 30.4% (w/w), to the load of Azythromycin up to 17.4% (w/w).
Four, satisfactory stability property: the prepared amphipathic amino acid block copolymer micelle of all-trans-retinoic acid, taxol, Zorubicin, cyclosporin A, Azythromycin among the embodiment; Be diluted to 1～3mg/ml; Place 4 ℃ and 25 ℃; The amphipathic amino acid block copolymer micelle of all-trans-retinoic acid stability difference＞5d, all the other 25 ℃ of stability are equal＞3d.It is thus clear that amphipathic amino acid block copolymer micelle not only has high drug loading, also has very superior stability.
Five, auxiliary material of the present invention also can be used in the blood vessel or intramuscular injection, oral, cavity or external application administration.This solid support material degradation in vivo is an amino acid, to human body safety.
In sum; The present invention adopts L-aspartic acid and the L-phenylalanine(Phe) of forming human body protein, synthesizing amphipathic amino acid segmented copolymer, spontaneous formation nano-micelle in water medium; Have excellent biological compatibility, biodegradability and stability in vivo, medicine is had load preferably.Amphipathic amino acid segmented copolymer spontaneous formation nano-micelle in water medium of the present invention's research, the bag that not only can be used for medicine carries, control drug release; And because the nano-micelle structure that its hydrophilic shell and hydrophobic nuclear are formed; Can the extension body internal recycle, reduce engulfing of reticuloendothelial cell, increase cancer target; Reach the minimizing toxic side effect, increase the purpose of curative effect.Amphipathic amino acid segmented copolymer micelle-forming concentration provided by the invention is low, drug loading is high, entrapment efficiency is high, nano-micelle is stable.This auxiliary material toxicity is little, can be used as the carrier of insoluble organic drug, is used in the blood vessel or intramuscular injection, oral, cavity or external application administration with the nano level dispersion system.Preparing method of the present invention is simple, and technical maturity is applicable to large scale continuous prod.
Description of drawings
Fig. 1: amphipathic amino acid segmented copolymer general formula
Fig. 2: SAP 73-β benzyl ester infared spectrum (embodiment 9)
Fig. 3: SAP 73-β benzyl ester
1H NMR collection of illustrative plates (embodiment 6)
Fig. 4: amphipathic amino acid segmented copolymer infared spectrum (embodiment 9)
Fig. 5: amphipathic amino acid segmented copolymer
1HNMR collection of illustrative plates (embodiment 6)
Fig. 6: the particle diameter collection of illustrative plates of amphipathic amino acid block copolymer micelle.(embodiment 10)
Fig. 7: the particle diameter collection of illustrative plates (embodiment 11) that is loaded with the amphipathic amino acid block copolymer micelle of all-trans-retinoic acid
Fig. 8: the Electronic Speculum collection of illustrative plates of amphipathic amino acid block copolymer micelle (embodiment 10)
Fig. 9: the Electronic Speculum collection of illustrative plates (embodiment 11) that is loaded with the amphipathic amino acid block copolymer micelle of all-trans-retinoic acid
Embodiment
Through embodiment the present invention is further explained below, but following embodiment does not limit to the interest field of this patent.
The preparation of embodiment 1:L-aspartic acid-beta benzyl ester ring inner-acid anhydride and L-phenylalanine(Phe) ring inner-acid anhydride
Reference (Prompruk K; Govender T; Zhang S; Et al.Synthesis of a novel PEG-block-poly (aspartic acid-statphenylalanine) copolymer shows potential for formation of a micellar drug carrier [J] .International Journal ofPharmaceutics, 2005,297 (1-2): 242-253.) carry out.Wherein L-aspartic acid-beta benzyl ester and L-phenylalanine(Phe) are available from Shanghai writing brush letter chemical inc, and TRIPHOSGENE 99.5 is available from the brilliant pure reagent in Shanghai ltd.
(1) L-phenylalanine(Phe) ring inner-acid anhydride preparation: the 10gL-phenylalanine(Phe) adds anhydrous tetrahydro furan 100ml, and magnetic stirs, and is warming up to 50 ℃; Add TRIPHOSGENE 99.5 15g then, suspension liquid becomes clarification back inflated with nitrogen 30min, to remove hydrogen chloride gas and the remaining phosgene that dereaction generates; Reaction solution is concentrated; Pour in the excessive anhydrous n-hexane, the sufficient crystallising after-filtration is with anhydrous n-hexane washing, the dry bullion that gets.Bullion is obtained the glossiness white needle-like crystals of bright sheet with anhydrous diethyl ether/normal hexane recrystallization be L-phenylalanine(Phe) ring inner-acid anhydride.Productive rate 85%.
Synthetic route is following:
(2) L-aspartic acid-beta benzyl ester ring inner-acid anhydride: 10gL-aspartic acid-beta-benzyl ester adds anhydrous tetrahydro furan 100ml, and magnetic stirs, and is warming up to 50 ℃; Add TRIPHOSGENE 99.5 10g then, suspension liquid becomes clarification back inflated with nitrogen 30min, to remove hydrogen chloride gas and the remaining phosgene that dereaction generates; Reaction solution is concentrated; Pour in the excessive anhydrous n-hexane, the sufficient crystallising after-filtration is with anhydrous n-hexane washing, the dry bullion that gets.Bullion is obtained the glossiness white needle-like crystals of bright sheet with anhydrous tetrahydro furan/normal hexane recrystallization be L-aspartic acid-beta benzyl ester ring inner-acid anhydride.Productive rate 89%.
Synthetic route is following:
Embodiment 2: the preparation of amphipathic amino acid segmented copolymer
10g L-aspartic acid-beta benzyl ester ring inner-acid anhydride adds Tri N-Propyl Amine 8mg stirring at room reaction 72h with N 20mL dissolving back, adds the absolute ethyl alcohol deposition, collecting precipitation; Take by weighing 1g after the vacuum-drying with N 50mL dissolving, add L-phenylalanine(Phe) ring inner-acid anhydride 0.95g again, room temperature continues stirring reaction 72h; Add ethanol sedimentation, collecting precipitation adds 2M NaOH 5mL stirring at room 4h after the vacuum-drying; Distill water dialysis 24h, lyophilize promptly gets.Productive rate 19.7%.
1HNMR measures; Through calculating the chemical shift peak area of aspartic acid section α precedence methyl proton; It is 183.0 that the chemical shift peak area of phenylalanine(Phe) section α precedence methyl proton is worth the aspartic acid polymerization degree with the chemical shift peak area ratio of holding methyl proton in the propyl group; The phenylalanine(Phe) polymerization degree is 144.1, and molecular weight is 46.3KDa.
Synthetic route is following:
Embodiment 3: the preparation of amphipathic amino acid segmented copolymer
10g L-aspartic acid-beta benzyl ester ring inner-acid anhydride adds n-Butyl Amine 99 500mg stirring at room reaction 12h with N 20mL dissolving back, adds the absolute ethyl alcohol deposition, collecting precipitation; Vacuum-drying takes by weighing 1g with N 10mL dissolving, adds L-phenylalanine(Phe) ring inner-acid anhydride 1.2g room temperature and continues stirring reaction 12h; Add the absolute ethyl alcohol deposition, collecting precipitation, vacuum-drying; Add 0.5M NaOH 5mL stirring at room 1h, distill water dialysis 24h, lyophilize promptly gets.Productive rate 45.6%.
1HNMR measures; Through calculating the chemical shift peak area of aspartic acid section α precedence methyl proton; It is 5.1 that the chemical shift peak area of phenylalanine(Phe) section α precedence methyl proton is worth the aspartic acid polymerization degree with the chemical shift peak area ratio of holding methyl proton in the propyl group; The phenylalanine(Phe) polymerization degree is 5.0, and molecular weight is 1.5KDa.
Embodiment 4: the preparation of amphipathic amino acid segmented copolymer
10g L-aspartic acid-beta benzyl ester ring inner-acid anhydride adds Tri N-Propyl Amine 150mg stirring at room reaction 24h with N 20mL dissolving back, adds the anhydrous diethyl ether deposition, collecting precipitation; Vacuum-drying takes by weighing 1g and adds 7g L-phenylalanine(Phe) ring inner-acid anhydride with N 50mL dissolving back, and room temperature continues stirring reaction 24h; Add the anhydrous diethyl ether deposition, collecting precipitation, vacuum-drying; Add 1M NaOH 5mL stirring at room 2h, distill water dialysis, lyophilize promptly gets.Productive rate 28.4%.
1HNMR measures; Through calculating the chemical shift peak area of aspartic acid section α precedence methyl proton; It is 12.5 that the chemical shift peak area of phenylalanine(Phe) section α precedence methyl proton is worth the aspartic acid polymerization degree with the chemical shift peak area ratio of holding methyl proton in the propyl group; The phenylalanine(Phe) polymerization degree is 54.3, and molecular weight is 9.8KDa.
Embodiment 5: the preparation of amphipathic amino acid segmented copolymer
Reaction conditions is with embodiment 4, and L-aspartic acid-beta benzyl ester ring inner-acid anhydride 10g dissolves with N 20mL, and Tri N-Propyl Amine 120mg takes by weighing 1g SAP 73-β benzyl ester and adds L-phenylalanine(Phe) ring inner-acid anhydride 0.9g with N 40mL dissolving back.Productive rate 43.2%.
1HNMR measures and calculates the aspartic acid polymerization degree is 15.4, and the phenylalanine(Phe) polymerization degree is 15.3, and molecular weight is 4.4KDa.
Embodiment 6: the preparation of amphipathic amino acid segmented copolymer
Reaction conditions is with embodiment 4, and L-aspartic acid-beta benzyl ester ring inner-acid anhydride 10g dissolves with N 20mL, Tri N-Propyl Amine 120mg, and making SAP 73-β benzyl polyisocyanate polyaddition degree is 15.6 (seeing accompanying drawing 3).Take by weighing 1g SAP 73-β benzyl ester with N 40mL dissolving back adding L-phenylalanine(Phe) ring inner-acid anhydride 1.4g, react amphipathic amino acid segmented copolymer (seeing accompanying drawing 5).Productive rate 37.6%.
1HNMR measures and calculates the aspartic acid polymerization degree is 15.6, and the phenylalanine(Phe) polymerization degree is 22.9, and molecular weight is 5.6KDa.
Embodiment 7: the preparation of amphipathic amino acid segmented copolymer
Reaction conditions is with embodiment 4, and L-aspartic acid-beta benzyl ester ring inner-acid anhydride 10g dissolves with N 20mL, and Tri N-Propyl Amine 30mg takes by weighing 1g SAP 73-β benzyl ester with add L-phenylalanine(Phe) ring inner-acid anhydride 0.12g with N 50mL dissolving back.Productive rate 42.6%.
1HNMR measures and calculates the aspartic acid polymerization degree is 49.7, and the phenylalanine(Phe) polymerization degree is 4.9, and molecular weight is 7.6KDa.
Embodiment 8: the preparation of amphipathic amino acid segmented copolymer
Reaction conditions is with embodiment 4, and aspartic acid-beta benzyl ester ring inner-acid anhydride 10g is with N 20mL dissolving, and Tri N-Propyl Amine 12mg takes by weighing 1g SAP 73-β benzyl ester and adds phenylalanine(Phe) ring inner-acid anhydride 20mg with N 50mL dissolving back.Productive rate 32.7%.
1HNMR measures and calculates the aspartic acid polymerization degree is 107.7, and the phenylalanine(Phe) polymerization degree is 2.4, and molecular weight is 15.2KDa.
Embodiment 9: the preparation of amphipathic amino acid segmented copolymer
10g L-phenylalanine(Phe) ring inner-acid anhydride adds Tri N-Propyl Amine 200mg stirring at room reaction 24h with N 20mL dissolving, adds the anhydrous diethyl ether deposition; Collecting precipitation is dry; Promptly get and gather L-phenylalanine(Phe) ring inner-acid anhydride (seeing accompanying drawing 2), take by weighing 1g and add L-aspartic acid-beta benzyl ester ring inner-acid anhydride 6.5g room temperature continuation reaction 36h, add the anhydrous diethyl ether deposition with N 50mL dissolving back; Collecting precipitation; Add 1MNaOH 5mL stirring at room 2h, distill water dialysis, lyophilize promptly gets (seeing accompanying drawing 4).Productive rate 31.8%.
1HNMR measures and calculates the aspartic acid polymerization degree is 33.7, and the phenylalanine(Phe) polymerization degree is 12.4, and molecular weight is 6.5KDa.
Synthetic route is following:
Embodiment 10: the preparation of amphipathic amino acid block copolymer micelle and sign
1. the preparation of amphipathic amino acid block copolymer micelle: amphipathic amino acid segmented copolymer 60mg, be dissolved in the 10ml water, stirring at room 20 minutes, then under the ice bath ultrasonic or high-pressure homogeneous after, 0.8 μ m membrane filtration promptly gets.
2. particle diameter: (UK) at 532nm, 25 ℃ of working sample particle diameters, the result sees table 1 to Zetasizer Nano ZS-90 for Malvern Instruments, Malvern.
3. micelle-forming concentration (critical micelle concentration; CMC): hydrophobicity fluorescent substance pyrene is very responsive to environment polarity; When amphiphilic materials forms micella in solution; Pyrene in the aqueous solution gets into micella kernel nonpolar environment, and then can observe a series of variations of fluorescence Spectra, wherein passes through with the I in the emmission spectrum of pyrene
1/ I
3(I
1, I
3Represent respectively pyrene when the fixed excitation wavelength scans first and the fluorescence intensity ratio at three strongest ones peak) or excitation spectrum in I
338/ I
333Can measure the CMC value than (the excitation spectrum medium wavelength is respectively the fluorescence intensity ratio of 338nm, 333nm) to the concentration logarithm mapping of amphipathic molecule, the result sees table 1.
The sign of the amphipathic amino acid block copolymer micelle of table 1
Embodiment 11:
The preparation and the sign that comprise the amphipathic amino acid block copolymer micelle of all-trans-retinoic acid
1. preparation technology
(1) the amphipathic amino acid segmented copolymer of ice-bath ultrasonic method 60mg is dissolved in the 10ml water, stirring at room 20 minutes.All-trans-retinoic acid 30mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; After the ice-bath ultrasonic 30 minutes, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm 10min; With 0.8 μ m membrane filtration, lyophilize.
(2) the amphipathic amino acid segmented copolymer of high pressure homogenization method 60mg is dissolved in the 10ml water, stirring at room 20 minutes.All-trans-retinoic acid 30mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; High-pressure homogeneous, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm 10min; With 0.8 μ m membrane filtration, lyophilize.
(3) the amphipathic amino acid segmented copolymer of solvent evaporation method 60mg is dissolved in the 10ml water, stirring at room 20 minutes.In the all-trans-retinoic acid 30mg dissolving chloroform, slowly drop in the amphipathic amino acid block copolymer solution, ultrasonic 30 minutes, continue stirred overnight, centrifugal 3000rpm 10min is with 0.8 μ m membrane filtration, lyophilize.
2. all-trans-retinoic acid Determination on content in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: methyl alcohol: water=92: 8 (v/v), flow velocity are 1.0mL/min, chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV, wavelength 345nm, 30 ℃ of column temperatures, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
3.Zetasizer (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for Nano ZS-90.
4. study on the stability
Freeze-drying samples using saline water is diluted to 1mg/kg, places 4 ℃ to investigate its stability.
Table 2 is loaded with the sign of the amphipathic amino acid block copolymer micelle of all-trans-retinoic acid
Embodiment 12: the preparation and the sign that comprise the amphipathic amino acid block copolymer micelle of taxol
1. preparation technology
Amphipathic amino acid segmented copolymer 60mg is dissolved in the 10ml water, stirring at room 20 minutes.Taxol 30mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; After the ice-bath ultrasonic 30 minutes, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm10min; With 0.8 μ m membrane filtration, lyophilize.
2. the assay of taxol in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: methyl alcohol: water=75: 25 (v/v), flow velocity are 1.0mL/min, chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV, wavelength 227nm, 30 ℃ of column temperatures, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
3.ZetasizerNano (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for ZS-90.
4. study on the stability
Freeze-drying samples using saline water is diluted to 3mg/kg, places room temperature to investigate its stability.
Table 3 is loaded with the sign of the amphipathic amino acid block copolymer micelle of taxol
Embodiment 13: the preparation and the sign that comprise the amphipathic amino acid block copolymer micelle of Zorubicin
1. preparation technology
Amphipathic amino acid segmented copolymer 10mg is dissolved in the 10ml water, stirring at room 20 minutes.Zorubicin 5mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop to amphipathic amino acid segmented copolymer; After the ice-bath ultrasonic 30 minutes, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm 10min; With 0.8 μ m membrane filtration, lyophilize.
2. the assay of Zorubicin in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: methyl alcohol: sodium-acetate buffer (Ph4.5)=70: 30 (v/v), flow velocity is 1.0mL/min, chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV, wavelength 233nm, room temperature, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
3.Zetasizer (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for Nano ZS-90.
4. study on the stability
Freeze-drying samples using saline water is diluted to 1mg/kg, places room temperature to investigate its stability.
Table 4 is loaded with the sign of the amphipathic amino acid block copolymer micelle of Zorubicin
Embodiment 14: the preparation and the sign that comprise the amphipathic amino acid block copolymer micelle of ciclosporin A
1. preparation technology
Amphipathic amino acid segmented copolymer 60mg is dissolved in the 3ml water, stirring at room 20 minutes.Ciclosporin A 30mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; After the ice-bath ultrasonic 30 minutes, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm10min; With 0.8 μ m membrane filtration, lyophilize.
2. the assay of ciclosporin A in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: acetonitrile: methyl alcohol: water: Virahol=900: 450: 50: 0.5 (v/v), flow velocity are 1.0mL/min, chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV, wavelength 225nm, 70 ℃ of column temperatures, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
3.ZetasizerNano (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for ZS-90.
4. study on the stability
Freeze-drying samples using saline water is diluted to 1mg/kg, places room temperature to investigate its stability.
Table 5 is loaded with the sign of the amphipathic amino acid block copolymer micelle of ciclosporin A
Embodiment 15: the preparation and the sign that comprise the amphipathic amino acid block copolymer micelle of Azythromycin
1. preparation technology
Amphipathic amino acid segmented copolymer 60mg is dissolved in the 10ml water, stirring at room 20 minutes.Azythromycin 30mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; After the ice-bath ultrasonic 30 minutes, steam with dialysis tubing (MWCO3500) room temperature dialysis 12h or pressurization in zero(ppm) water and to remove organic solvent, centrifugal 3000rpm10min; With 0.8 μ m membrane filtration, lyophilize.
2. the assay of taxol in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: acetonitrile: 0.1% triethylamine aqueous solution=30: 70 (v/v), transfer pH4.0 with phosphoric acid, flow velocity is 1.0mL/min; Chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV; Wavelength 205nm, 40 ℃ of column temperatures, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
3.Zetasizer (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for Nano ZS-90.
4. study on the stability
Freeze-drying samples using saline water is diluted to 1mg/kg, places room temperature to investigate its stability.
Table 6 is loaded with the sign of the amphipathic amino acid block copolymer micelle of Azythromycin
Embodiment 16: amphipathic amino acid segmented copolymer pH study on the stability
1. the preparation of amphipathic amino acid block copolymer micelle: take by weighing 10 parts of embodiment 9 amphipathic amino acid segmented copolymer 36mg; Be dissolved in respectively in 6ml pH3.6, Ph4.0, pH4.6, pH5.0, pH5.6, pH6.0, pH6.5, pH7.0, pH7.4, the pH8.0 damping fluid; Stirring at room 20 minutes; Then under the ice bath ultrasonic or high-pressure homogeneous after, 0.8 μ m membrane filtration promptly gets.
2. the ice-bath ultrasonic method takes by weighing embodiment 9 amphipathic amino acid segmented copolymer 36mg9 parts and is dissolved in 6mlpH3.6, Ph4.0, pH4.6, pH5.0, pH5.6, pH6.0, pH6.5, pH7.0, pH7.4, the pH8.0 damping fluid stirring at room 20 minutes.All-trans-retinoic acid 18mg is dissolved among the DMSO (methyl alcohol, ethanol, acetonitrile); Slowly drop in the amphipathic amino acid block copolymer solution; After the ice-bath ultrasonic 30 minutes, with dialysis tubing (MWCO3500) respectively in pH3.6, Ph4.0, pH4.6, pH5.0, pH5.6, pH6.0, pH6.5, pH7.0, pH7.4, pH8.0 damping fluid room temperature dialysis 12h or pressurization steam and remove organic solvent, centrifugal 3000rpm 10min; With 0.8 μ m membrane filtration, lyophilize.
3. all-trans-retinoic acid Determination on content in the amphipathic amino acid block copolymer micelle
(LC-20AT, Shimazu Japan) carry out assay with HPLC.Moving phase: methyl alcohol: water=92: 8 (v/v), flow velocity are 1.0mL/min, chromatographic column Hypersil ODS2 (4.6mm * 250mm, 5 μ m), detector SPD-20AV, wavelength 345nm, 30 ℃ of column temperatures, sampling volume 20 μ L.Drug loading with formula (1) calculation sample.
4.Zetasizer (Malvern Instruments, Malvern is UK) at 532nm, 25 ℃ of working sample particle diameters for Nano ZS-90.
5. study on the stability
Freeze-drying samples using saline water is diluted to 1mg/kg, places 4 ℃ to investigate its stability.
Amphipathic amino acid segmented copolymer pH study on the stability result sees table 7.
The amphipathic amino acid segmented copolymer of table 7 pH study on the stability
Claims (8)
1. general formula is formula (I)
Amphipathic amino acid segmented copolymer, wherein, the positive integer of m=1～200; The positive integer of n=1～150; X is an active end group ammonia, and Y is a basic metal.
2. the described amphipathic amino acid segmented copolymer of claim 1 is characterized in that: the positive integer of m=5～50, and the positive integer of n=5～50, m is 1: 5～5: 1 with the ratio of n; Its molecular weight is 1KDa～50KDa.
3. the described amphipathic amino acid segmented copolymer of claim 1 is characterized in that said X is Tri N-Propyl Amine, n-Butyl Amine 99, n-amylamine or normal hexyl Amine.
4. the preparation method of the described amphipathic amino acid segmented copolymer of claim 1, this method comprises: L-aspartic acid-beta benzyl ester ring inner-acid anhydride is added active end group ammonia with N dissolving back, and room temperature condition stirs 12～72h down; Add anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product, collecting precipitation, dissolve with N dry back; Add L-phenylalanine(Phe) ring inner-acid anhydride again, room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product; Collecting precipitation, drying, adding concentration is the NaOH solution of 0.5～2M; Stirring at room 1～4h, solution promptly gets formula (I) segmented copolymer through the dialysis tubing after drying of dialysing; Or earlier L-phenylalanine(Phe) ring inner-acid anhydride being added active end group ammonia with N dissolving back, room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product; Collecting precipitation, dry back adds L-aspartic acid-beta benzyl ester ring inner-acid anhydride again with the N dissolving; Room temperature condition stirs 12～72h down, adds anhydrous diethyl ether or absolute ethyl alcohol precipitin reaction product, collecting precipitation; Drying, adding concentration is the NaOH solution of 0.5～2M, stirring at room 1～4h; Solution is dialysed through dialysis tubing, and drying promptly gets formula (II) segmented copolymer.
5. the described amphipathic amino acid segmented copolymer of claim 1 is used in medicament solubilization.
6. the described application of claim 5 is characterized in that described medicine is selected from: retinoid, taxanes, camptothecin, ciclosporin class, flavonoid 、 Xiao ? arbitrary material or derivatives thereof in bases, dihydropyridines, vinca, podophillotoxines, anthraquinone series antineoplastic medicament, steroidal class or nonsteroidal anti-inflammatory drug, microbiotic, cardiovascular agent, antiviral, antifungal drug, the immunomodulator.
7. the described application of claim 6 is characterized in that described medicine is selected from: all-trans-retinoic acid, taxol, Azythromycin, Zorubicin or ciclosporin A.
8. application according to claim 5; The preparation method who it is characterized in that this amphipathic amino acid segmented copolymer medicament solubilization carrier may further comprise the steps: dissolve by weight the ratio that is 1～50: 1000 with water, obtain the nano-micelle of amphipathic amino acid segmented copolymer; With the indissoluble of treatment significant quantity or the organic drug that is slightly soluble in water with after the acceptable solvent dissolving pharmaceutically; After said amphipathic amino acid block copolymer nano micella mixes; Handle through ultrasonic or high pressure homogenization; Solvent is removed organic solvent and small molecules with dialysis tubing dialysis or ultrafiltration process or post partition method, and drying makes the nano-micelle that particle diameter is 10～1000nm.
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| CN108079307A (en) * | 2018-02-08 | 2018-05-29 | 中国药科大学 | A kind of tri compound nanometer system and its application based on methoxy polyethylene glycol-polylactic acid |
| CN110652595A (en) * | 2019-10-21 | 2020-01-07 | 南通大学附属医院 | Poly leucine-poly aspartic acid segmented copolymer stereo composite drug-loaded micelle and preparation method thereof |
| CN112430320A (en) * | 2020-11-25 | 2021-03-02 | 温州医科大学 | Multi-hydrophobic core side chain polymer, multi-hydrophobic core drug-loaded material and FK506 preparation |
| CN115475271A (en) * | 2022-08-19 | 2022-12-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of amino acid/rare earth nanocrystalline/nanocellulose antibacterial hemostatic dressing |
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2011
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108079307A (en) * | 2018-02-08 | 2018-05-29 | 中国药科大学 | A kind of tri compound nanometer system and its application based on methoxy polyethylene glycol-polylactic acid |
| CN110652595A (en) * | 2019-10-21 | 2020-01-07 | 南通大学附属医院 | Poly leucine-poly aspartic acid segmented copolymer stereo composite drug-loaded micelle and preparation method thereof |
| CN112430320A (en) * | 2020-11-25 | 2021-03-02 | 温州医科大学 | Multi-hydrophobic core side chain polymer, multi-hydrophobic core drug-loaded material and FK506 preparation |
| CN112430320B (en) * | 2020-11-25 | 2023-01-31 | 温州医科大学 | Multi-hydrophobic core side chain polymer, multi-hydrophobic core drug-loaded material and FK506 preparation |
| CN115475271A (en) * | 2022-08-19 | 2022-12-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of amino acid/rare earth nanocrystalline/nanocellulose antibacterial hemostatic dressing |
| CN115475271B (en) * | 2022-08-19 | 2023-11-24 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of amino acid/rare earth nanocrystalline/nanocellulose antibacterial hemostatic dressing |
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