WO2006015534A1 - Procede de preparation des nanoparticules doxorubicine-polybutylcyanoacrylate - Google Patents

Procede de preparation des nanoparticules doxorubicine-polybutylcyanoacrylate Download PDF

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WO2006015534A1
WO2006015534A1 PCT/CN2005/001056 CN2005001056W WO2006015534A1 WO 2006015534 A1 WO2006015534 A1 WO 2006015534A1 CN 2005001056 W CN2005001056 W CN 2005001056W WO 2006015534 A1 WO2006015534 A1 WO 2006015534A1
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pbca
doxorubicin
drug
stir
dox
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Yangde Zhang
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Yangde Zhang
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/513—Organic macromolecular compounds; Dendrimers
    • A61K9/5138—Organic macromolecular compounds; Dendrimers obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/58—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to a method for preparing nanometer particles for treating liver cancer.
  • liver cancer is one of the most common malignant tumors in the world. At present, the treatment of liver cancer is still clinically based on a comprehensive treatment plan, and many cases still require chemotherapy. However, the toxic side effects of chemotherapy cause loss of treatment opportunities for patients with poor liver function and general condition. In recent years, gene therapy has been produced and developed, but the premise of effective gene therapy is that nucleic acid molecules must be efficiently delivered to target cells.
  • DOX-PBCA-NP is: using HCl solution and doxorubicin hydrochloride powder to stir and dissolve, then adding DextranTM, adding n-butyl ⁇ -cyanoacrylate, stirring to be orange-yellow milk,
  • the present invention was prepared by adjusting ⁇ 2, passing through a 0.54 ⁇ filter, and drying.
  • the DOX-PBCA-NP obtained by the invention can carry a plurality of drugs including chemotherapeutic drugs and gene drugs, and the targeting performance thereof can significantly reduce the toxicity of the chemotherapeutic drugs, improve the curative effect, reduce the dosage of the drugs and the number of drugs, and improve the patient. Compliance, improve the stability of genetic drugs on ribozymes, prolong biological half-life, increase intracellular drug concentration and duration of action.
  • the invention first prepares blank PBCA nanoparticles, and then screens the factors affecting the preparation of PBCA nanoparticles encapsulating doxorubicin, and optimizes the preparation process by using the homogenization design, and further improves the drug content by improving the interaction between the model drug and the carrier.
  • the complex of doxorubicin and plasmid p53 gene was simultaneously encapsulated by the amphipathic reagent CTAB.
  • the 5% Dextran 7 was added to the micropipette.
  • the solution was added to the solution.
  • the solution was added to the solution.
  • BCA n-butyl cyanoacrylate
  • Dextran70 slowly add ⁇ -cyanoacrylate n-butyl ester (BCA) under magnetic stirring 0. 2ml in the above solution (about lOmin added), while stirring, add dropwise, close the bottle mouth, continue to stir lOOOrmp * 3h, 0, ⁇ 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 0. 45 ⁇ ⁇ filter.
  • BCA ⁇ -cyanoacrylate n-butyl ester
  • DOX-PBCA-NP was prepared by one-step and two-step methods, and there was no significant difference in ER and LD between the two groups.
  • the ER of DOX-PBCA-NP increased with the increase of BCA monomer in the prescription, and the ER of DOX-PBCA-NP increased gradually (P ⁇ 0.05), while its LD Then it decreased with the increase of BCA monomer (P ⁇ 0.05).
  • the amount of BCA in the material increased from 0.1ml to 0.4ml.
  • the encapsulation efficiency of the nanoparticles increased from 46.59% to 96.25%, which increased by 1.07 times, and the loss of D0X decreased from 53.41% to 3.75%.
  • the drug loading amount was from 4.27%. It fell to 2.58%, only 0.53.
  • the amount of BCA monomer is increased, the particle size is also increased and the distribution is uneven. See Table 3 and Figure 3, Figure 4 and Figure 9.
  • the content of D0X - PBCA- NP drug did not change significantly with the increase of stirring speed, and there was no difference between the groups.
  • the zeta potential is negative, that is, the PBCA-NP after encapsulation of doxorubicin is still negatively charged. See Table 8 and Figure 5, Figure 6.
  • Figure 1 is a graphical representation of the effect of doxorubicin hydrochloride on DOX-PBCA-NP ER.
  • Figure 2 is a graphical representation of the effect of doxorubicin hydrochloride on D0X-PBCA-NP LD.
  • Figure 3 is a graphical representation of the effect of BCA dosage on the encapsulation efficiency of PBCA-NP doxorubicin.
  • Figure 4 is a graphical representation of the effect of BCA dosage on PBCA-NP doxorubicin drug loading.
  • Figure 5 is the relationship between the ER of DOX-PBCA-NP and its zeta potential after the addition of N3 ⁇ 4S0 4 .
  • Figure 6 shows the relationship between LD and its Zeta potential added to N3 ⁇ 4S0 ⁇ D0X-PBCA-NP.
  • Figure 7 is a two-step nanoparticle TEM (100000X).
  • Figure 8 is a one-step nanoparticle TEM (100000X).
  • Figure 10 is a prepared nanoparticle TEM (40000X).
  • Figure 11 is a prepared nanoparticle SEM (60000X).

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  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Description

阿霉素一聚氰基丙烯酸正丁酯纳米粒的制备方法 技术领域
本发明涉及一种对肝癌治疗药物纳米颗粒的制备方法。
背景技术
. 肝癌是世界最常见的恶性肿瘤之一， 目前， 治疗肝癌临床上仍采取手术切 除为主的综合治疗方案，许多病例仍需接受化疗。但化疗的毒副反应使肝功能 及全身情况不佳者丧失治疗机会。近年来， 产生和发展了基因治疗， 但有效基 因治疗的前提是必须将核酸分子有效递送到靶细胞中。
发明内容
本发明的目的，是要提供一种具有较强的靶向性的阿霉素一聚氰基丙烯酸 正丁酯纳米粒 （DOX PBCA-NP) 的制备方法。
本发明 DOX- PBCA-NP的制备方法是：用 HC1溶液与盐酸阿霉素粉末作用搅拌 使其溶解， 再加入 Dextran™, 加入 α—氰基丙烯酸正丁酯， 搅拌待出现橙黄色 乳光， 调节 ΡΗ二 7, 过 0. 45 μ ηι滤膜， 干燥， 即制得本发明。
用本发明获得的 DOX- PBCA-NP, 可运载包括化疗药物、 基因药物在内的多 种药物， 其靶向性能显著降低化疗药物的毒性， 提高疗效， 能减少药物用量及 用药次数，提高患者依从性，提高基因药物对核酶的稳定性，延长生物半衰期， 提高胞内药物浓度和作用时间。
本发明先制备出空白 PBCA纳米粒，然后筛选影响包封阿霉素的 PBCA纳米粒 制备的诸因素，采用均勾设计优化制备工艺， 从改善模型药物与载体的相互作 用进一步提高其药物含量， 并通过双亲性试剂 CTAB同时包封阿霉素和质粒 p53 基因的复合物。
单因素初选
1、 空白 PBCA— NP的制备
移液管移取 0. IN HC1溶液加入锥形瓶中， 微量移液器加入 1. 5%DeXtran7。， 磁力搅拌下缓慢加入 α—氰基丙烯酸正丁酯（BCA) 0. 2ml于上述溶液（约 lOmin 加完） ， 边滴加边搅拌， 滴加完毕， 封闭瓶口， 持续搅拌 1000rpm*3h， 体系出 现白色乳光时终止反应， 0. 2N NA0H溶液调节 1¾=7. 0， 过 0. 45 μ πι滤头。 取少 量滤液于真空干燥箱中 25° (：、一 0. lkPa条件下干燥，样品送电镜室观察粒径粒 形， 余备用。
2、 不同制备工艺对 DOX- PBCA- NP药物含量的影响。
2. 1 一步法
移液管移取 0. 1N HC1溶液加入锥形瓶中，取干燥至恒重的盐酸阿霉素粉末 10. OOmg于上述溶液， 搅拌至溶，微量移液器加入 1. 5%Dextran7。， 磁力搅拌下 缓慢加入 α—氰基丙烯酸正丁酯 （BCA) 0. 2ml于上述溶液 （约 lOmin加完） ， 边滴边搅拌， 滴加完毕， 封闭瓶口， 持续搅拌 1000rmp*3h， 体系出现橙黄色乳 光时终止反应， 0. 2N NaOH溶液调节 PH=7. 0， 过 0. 45 μ πι滤头。
2. 2二步法
移液管移取 0. IN HC1溶液加入锥形瓶中， 微量移液器加入 1. 5%
Dextran70, 磁力搅拌下缓慢加入 α—氰基丙烯酸正丁酯 （BCA) 0. 2ml于上述 溶液（约 lOmin加完） ， 边滴边搅拌， 滴加完毕， 封闭瓶口， 持续搅拌 lOOOrmp *3h，体系出现白色乳光时终止反应，取干燥至恒重的盐酸阿霉素粉末 10. OOmg 于上述溶液， 搅拌至溶， 继续搅拌 2h, 0. 2N NaOH溶液调节 PH=7. 0， 过 0. 45 μ πι 滤头。
各种相关因素对药物含量的影响：
1、 制备方法对 DOX- PBCA-NP药物含量的影响
采用一步法和二步法制备 DOX- PBCA-NP, 两组间 ER和 LD差异无显著性
(Ρ>0. 05) ， 见表 1， 但一步法制备的纳米粒粒径较二步法大。 如图 7、 图 8。
制备工艺对 DOX- PBCA- NP ER和 LD的影响 （η=3)
制备工艺 ER(%) LD (%)
一步法 65. 22+/- 1. 64 3. 46+/- 0. 09
二步法 63. 54+/-0. 76# 3. 42+/- 0. 04#
注： #相邻两组间相比 P〉0. 05。
2、 盐酸阿霉素用量对 D0X-PBCA-NP药物含量的影响 依 2.1处方和工艺， DOX-PBCA- NP的 ER随着处方中的阿霉素的用量的增加而 逐渐降低， 差异有显著性（P<0.05或 P<0.01) ， 而其载药量则随着处方中阿霉 素的增加而增加， 5.0mg组低于其它各组 (P<0.05) ， 但 20. Omg组与 15. Omg组 的载药量无显著性差异 （P〉0.05) 。 如表 2和图 1、 图 2。
表 2 盐酸阿霉素用量对 DOX-PBCA-NP药物含量的影响 （n=3)
DOX量 （mg) ER(%)
Figure imgf000005_0001
5.0 76.80+/-0.02 2.11+/—0.05
10.0 65.22+/- 1.64* 3.46+/- 0, 09*
15.0 44.89+/- 0.36* 3.70+/- 0.03*
20.0 33· 76+/-0.46* 3.71+/-0.05#
注： *表示相邻组相比 Ρ<0.05， #表示相邻两组间相比 Ρ>0.05。
3、 BCA单体用量对 DOX- PBCA-NP药物用量的影响
参照 2.1工艺，改变处方中 BCA单体的用量后， DOX-PBCA-NP的 ER随着处方中 的 BCA单体的增加， DOX- PBCA-NP的 ER逐渐增加 （P〈0.05) ，而其 LD则随 BCA单体 的增加而降低（P<0.05) 。材料 BCA量从 0.1ml增至 0.4ml， 纳米粒的包封率相应 从 46.59%增至 96.25%， 增加 1.07倍， D0X的损耗则由 53.41%降至 3.75%; 而 载药量则从 4.27%降至 2.58%, 仅为原来的 0.53。 BCA单体用量增加的同时其 粒径也增大， 分布不均。 如表 3和图 3、 图 4及图 9。
表 3 BCA单体用量对 DOX- PBCA-NP药物含量的影响 （n=3)
BCA量 （ml) ER(%) LD(%)
0.1 46.59+/— 1.28 4.87+/— 0.14
0.2 65.22+/- 1.64* 3.46+/- 0.09*
0.3 87.05+/-2.46* 3· 09+/- 0.07*
0.4 96.25+/- 0.38* 2.58+/- 0.01*
注： *表示相邻组相比 Ρ<0.05
4、 值对 DOX-PBCA- NP药物含量的影响
BCA单体 0.2ml， D0X10. Omg, 依 2.1工艺制备 DOX- PBCA-NP, ER随着 PH值的 增大而逐渐减小， 但 PH=2， 3两组的 ER与 PH二 1时的 ER相比， 差异无显著性
(P>0. 05 ) ， 当 PH值增至 5以上时， ER明显减小， PH=5， 6组与 PH=1组相比差异 有显著性 (P<0. 05) ， 载药量在 PH二 2， 3， 5时虽然减小， 但与 PH=1时相比差异 不显著 （P>0. 05 ) 。 见表 4。 另外， 体系出现乳光的时间随着 PH值的降低而逐 渐延长， PH=1时反应时间长达 6- 7小时。 还发现当体系 PH值〉 8时制备的纳米粒 难以成球， 产品在镜下呈片状物， 见图 10。
表 4 PH值对 D0X- PBCA-NP药物含量的影响 （n = 3 )
Figure imgf000006_0001
1. 0 65. 42+/- 3. 10 3. 46+/- 0. 17
2. 0 65. 22+/-1. 64 3. 46+/- 0. 09
3. 0 65. 22+/— 2. 00 3. 46+/-0. 11
5. 0 63. 86+/- 3. 65* 3. 39+/- 0. 20
6. 0 54. 74+/-4. 38* 2. 92+/- 0. 24*
注： *表示与 PH=1. 0组相比 P〈0. 05
5、 搅拌速度对 DOX- PBCA-NP药物含量的影响
D0X - PBCA- NP药物含量随着搅拌速度的增加而变化不明显， 各组间无差异
(P>0. 05 ) ，唯粒经分布随着搅拌速度增大而增宽。 见表 5
表 5 搅拌速对 DOX- PBCA- NP药物含量的影响 （n=3 )
搅拌速度 （rpm) ER (%) LD (%)
1000 65. 22+/- 1. 64 3. 46+/-0. 09
2000 65. 29+/-1. 26# 3. 46+/-0. 06#
3000 65. 31+/- 0. 78# 3. 47+/- 0. 04*
注： #表示与搅拌速度 = 1000rpm组相比 P>0. 05。
6、 均匀设计安排结果
依均匀设计方案实验， 电镜下评估纳米粒外观（粒径、 粒形） ， 分光光度 法测定阿霉素包封率， 计算载药量， 结果如表 6。 经多元回归分析处理， 得多 元线性回归方程： Y=19. 456+0. 113¾-1. 66 - 6. 121¾- 0. 839 ， F二 0. 888, R-0. 686, R2二 0. 57, Ρ二 0. 544。 由方程可见， ¾、 、 系数均为负值， 表明 其取值宜小， 而 系数为正值， 表明其取值宜适当的大。 结合单因素分析得结 果， PH=1时反应太慢，粒径分布较宽，综合考虑后的优化条件为： DOX-10. Omg， BCA单体 =0.25ml， PH=2.5, 稳定剂 = 1.5%， 按上述条件制备纳米粒， 结果包 封率为 79.31+/- 1.17%， 载药量为 3.49+/- 0.05%， 外观如图 11。
表 6 均匀设计的实验结果评分
Figure imgf000007_0001
7.5 8.21 0.148 15.858
4.0 5.73 0.087 10.817
7.0 7.55 0.318 14.686
3.0 3.24 0.092 6.332
7.5 5.49 0.418 13.408
8.6 8.26 0.389 17.249
7.8 4.07 0.763 12.733
7.5 4.31 0.317 14.947
4.0 1.98 0.103 6.083
7.电解质对 DOX- PBCA-NP药物含量的影响
随着处方中三种电解质加入， 各试验组的 D0X药物含量均有所增加， 但只 有硫酸钠组与对照组相比包封率和载药量增加的差异有显著性 （P〈0.05) ，而 NaCl, N C03组与对照组相比药物含量增加的差异性不显著（P>0.05)。如表 7。 实验过程中还发现，体系中加入上述剂量的电解质后，各实验组与对照组均未 见明显的肉眼观絮凝和聚结现象发生。
表 7 电解质对 DOX- PBCA- NP药物含量的影响 （n=3)
电解质 ER(%) LD(%)
none 82.31+/- 1.03 3.49+/- 0.05
NaCl 82.70+/- 0.90 3.50+/-0.04
Na2C03 83.83+/-0.34 3.55+/-0.01
N¾S04 84.26+/-0.27* 3.57+/- 0.01*
注： *表示与对照组相比 P〈0.05
8.不同浓度的 Na2S0对 D0X-PBCA-NP药物含量的影响 加入 Na2S04后， DOX- PBCA- NP的 ER和 LD增加， 8.0， 12· Omg/ml二个剂量水平 组包封率和载药量均较未加 N S04组增高， 差异有显著性（P〈0.05) ， 但 N S04 达 16. Omg/ml时药物含量较对照组并未增加，差异无显著性 (P〉0.05), 4. Omg/ml 水平组与对照组相比差异无显著性（P>0.05)，对应的 Zeta电位亦有相同改变。 N S04的剂量水平在 0mg/ml〜16. Omg/ml时，纳米粒的 Zeta电位的大小与 ER和 LD 呈高度的负相关 （rER=0.9671， rLD=0.9125, P<0.05) 。 其 Zeta电位均为负值， 即包封阿霉素后的 PBCA- NP仍然带负电。 如表 8和图 5、 图 6。
表 8 不同浓度的 N¾S0^ D0X- PBCA-NP药物含量的影响 （n=3)
Na2S04 (mg/ ½1) 0 4.0 8.0 12.0 16.0
ER(%) 79.31+/-1.17 83.11+/-1.12 83.90+/- 3.03* 84.25+/- 4.19* 80.00+/- 2.11
LD(%) 3.49+/- 0.05 3.53+/- 0.04 3.56+/- 0.13* 3.57+/-0.18* 3.40+/- 0.09
Zeta (mv) -18.3+/-0.9 -26.7+/- 1.2 -31.4+/-1.8* -34.5+/-1.3* -16.6+/-1.7 注： *表示与对照组相比 P<0.05
9、 离子对试剂对 DOX- PBCA- NP药物含量的影响
随着含羧基 C00-化合物油酸钠的加入， 6.0， 9. Omg/ml组与对照组相比包 封率、 载药量增加， 差异有显著性（P〈0.05) ， 但 3. Omg/ml组与对照组相比差 异无显著性 （P>0.05) 。 见表 9
表 9 离子对试剂对 DOX- PBCA- NP药物含量的影响 （n二 3)
油酸钠（mg/ml) 0 3.0 6.0 9.0
ER(%) 79.31+/ - 1.17 82.70+/ - 3.32 85.68+/-1.40* 87. Θ8+/-2.55*
LD(%) 3.49+/-0.05 3.59+/-0.14 3.63+/- 0.06* 3.71+/-0.11* 注: *表示与对照组相比 P<0.05
10、 p53- DOX- PBCA-纳米粒的药物含量
加入季胺盐 CTAB后， p53的包封率明显提高， 差异有显著性（P〈0.05)，而阿 霉素的包封率变化不大， 与对照组比较差异无显著性 （P〉0.05) 。 见表 10 表 10 p53- DOX- PBCA-纳米粒的包封率 （n=3)
CTAB量 （ μ M) 0 500.0
DOX 82.33+/- 0.09 82.17+/-0.22
ER(%)
p53 23.86+/- 0.28 49.67+/-1.26*
注： *表示与对照组相比 P<0.05
附图说明
图 1是盐酸阿霉素用量对 DOX-PBCA-NP ER的影响图示。
图 2是盐酸阿霉素用量对 D0X-PBCA- NP LD的影响图示。
图 3是 BCA用量对 PBCA- NP阿霉素包封率的影响图示。
图 4是 BCA用量对 PBCA-NP阿霉素载药量的影响图示。
图 5是加入 N¾S04后 DOX- PBCA-NP的 ER与其 Zeta电位的关系。
图 6是加入 N¾S0^D0X- PBCA-NP的 LD与其 Zeta电位的关系。
图 7是二步法所得纳米粒 TEM (100000X) 。
图 8是一步法所得纳米粒 TEM (100000X) 。
图 9是 BCA单体 =0.60ml时所得纳米粒 TEM (35000X) 。
图 10是制备的纳米粒 TEM (40000X) 。
图 11是制备的纳米粒 SEM (60000X) 。
具体实施方式
下面结合实施例具体介绍本发明。
用移液管取 0.1NHC1溶液 50ml置于锥形瓶中， 加入干燥至恒重的盐酸阿霉 素粉末 10mg， 搅拌使其溶解， 再用微量移液器加入1.5%06^^117。0.31111，磁力 搅拌器作用下， 缓慢加入 α—氰基丙烯酸正丁酯 0.2ml， 边滴加边搅拌， 控制 在 10分钟加完。 封闭瓶口， 持续搅拌 1000rpm3h， 待出现橙黄色乳光， 用 0.2N NaOH溶液调节 PH=7， 过 0.45μπι滤膜。 滤液于真空干燥箱中 25°C、 0. lkPa条件 下干燥， 即制得本发明。

Claims

权 利 要 求
1、 一种阿霉素一聚氰基丙烯酸正丁酯纳米粒的制备方法， 其步骤是： 用 HC1溶液与盐酸阿霉素粉末作用， 搅拌使其溶解， 再加入 Dextran™, 加入 α— 氰基丙烯酸正丁酯， 搅拌待出现橙黄色乳光， 调节 ΡΗ=7， 过 0.45μπι滤膜， 干 燥， 即制得本发明。
2、 按权利要求 1所述的阿霉素一聚氰基丙烯酸正丁酯纳米粒的制备方法， 设制取一份 D0X— PBCA— ΝΡ， 其步骤是： 用移液管取 0.1N HC1溶液 50ml， 置于 锥形瓶中， 加入干燥至恒重的盐酸阿霉素粉末 10mg， 搅拌使其溶解， 再用微量 移液器加入1.5%0^1^&117。0.31111，磁力搅拌器作用下，缓慢加入 α—氰基丙烯 酸正丁酯 0.2ml， 边滴加边搅拌， 控制在 10分钟加完； 封闭瓶口， 持续搅拌 lOOOrpm 3h, 待出现橙黄色乳光， 用 0.2NNaOH溶液调节 PH二 7， 过 0.45μπι滤膜， 滤液于真空干燥箱中 25°C、 一 O.lkPa条件下干燥， 即制得本发明。
PCT/CN2005/001056 2004-08-11 2005-07-18 Procede de preparation des nanoparticules doxorubicine-polybutylcyanoacrylate WO2006015534A1 (fr)

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CN115444810B (zh) * 2022-04-27 2023-10-13 贵州医科大学 一种载阿霉素纳米粒复合温敏凝胶的制备方法

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