【推荐文章】光敏感型盐酸多柔比星脂质体逆转乳腺癌耐药

   本文旨在构建一种新型光敏感共输送脂质体, 联合光动力学治疗与化疗, 逆转乳腺癌耐药。采用薄膜水化挤出−硫酸铵主动载药法制备共输送光敏剂二氢卟吩e6三甲酯 (chlorin e6 trimethyl ester, Ce6tM) 与盐酸多柔比星 (doxorubicin hydrochloride, DOX) 的脂质体 (liposomes loaded with Ce6tM and DOX, CDL)。结果表明, CDL具有良好的光敏感释药特性，这种光敏感释药行为有助于增加DOX在MCF7/ADR细胞中的蓄积。药效学实验结果表明, 光敏感CDL在逆转乳腺癌耐药治疗方面具有明显的潜在应用价值。

作者简介——《药学学报》英文刊编委

李亚平，中科院上海药物研究所研究员，国家有突出贡献中青年专家，杰青，中科院百人计划；近5年，在国际重要学术期刊上发表90余篇高质量SCI论文（IF>10的论文36篇）；已获得新药证书9本，临床批件6件。

【研究方向】主要从事基于纳米技术的药物靶向递释系统和高端制剂研究开发

【在《药学学报》发表论文】光敏感型盐酸多柔比星脂质体逆转乳腺癌耐药

Part
1

 

 CDL处方的筛选与表征

Figure 1  Formulation screening of liposomes loaded with chlorin e6 trimethyl ester (Ce6tM) and doxorubicin hydrochloride (DOX) (CDL).  Percentages of DOX release from different CDL formulations after treatment with 6 h, 37 ℃ water incubation (A) or 1 min, 2 W·cm⁻², 671 nm laser irradiation (B); C: Cryo-transmission electron microscopy (Cryo-TEM) images of CDL; D: Particle size distribu­tion of CDL measured by dynamic light scattering.  SPC: Soybean phospholipid

Part
2

 

DOX的光敏感释放

光敏感释药机制

Figure 2  Photo-sensitive DOX release behaviors in vitro of CDL and its mechanism.  A: Cumulative release of DOX from CDL or liposomes loaded with DOX (SDL) in PBS (pH 7.4) under 671 nm laser irradiation with different power; B: Cumulative release of DOX from CDL or SDL after 15 s, 0.25 W·cm⁻², 671 nm laser irradiation in 37 ℃, PBS (pH 7.4); C, D: Cryo-TEM images and particle size distribution of CDL after 4 min, 2 W·cm⁻², 671 nm laser irradiation; E: Temperature change curves of CDL, SCL or SDL under 671 nm laser irradiation with different power; F: Cumulative release of DOX from CDL in 60 ℃ water bath; G: Cumulative release of DOX from CDL in 37 ℃ PBS (pH 7.4) after 15 s, 60 ℃ water incubation treatment; H: Generation of ¹O₂ by CDL or SDL under 671 nm laser irradiation; I: Detection of oxidation products malondialdehyde (MDA) from liposomes loaded with Ce6tM (SCL) after 671 nm laser irradiation; J: Effects of ¹O₂ quencher NaN₃ on DOX release rate from CDL under 671 nm laser irradiation (2 W·cm⁻²).  n=3, x±s

Part
3

 

DOX细胞内蓄积

Figure 3  DOX accumulation in MCF7/ADR cells after 671 nm laser irradiation (2 W·cm⁻², 2 min).  A: Quantitative analysis of DOX accumulation in MCF7/ADR cells after treatment with Free DOX, SDL, CDL, or SDL SCL mixture for 1, 2, 4, 8, 12 h with or without 671 nm laser irradiation (2 W·cm⁻², 2 min).  Laser irradiation was executed at 12 h before detection; B, C: Fluorescence images of DOX or Ce6tM in MCF7/ADR cells after incubation with Free DOX, SCL, SDL, CDL, or SDL SCL mixture for 12 h before or after 671 nm laser irradiation (2 W·cm⁻², 2 min).  Laser irradiation was executed at 12 h.  Scale bars represented a distance of 200 μm.  n=3, x±s.  ^***P<0.001

Figure 4  DOX accumulation in MCF7/ADR cells after 671 nm laser irradiation (0.25 W·cm⁻²,15 s).  A: Fluorescence images of DOX in MCF7/ADR cells after incubation with CDL for 12 h or 18 h with or without 671 nm laser irradiation (0.25 W·cm⁻², 15 s).  Laser irradiation was executed at 12 h.  Scale bars represented a distance of 200 μm.  B: Quantitative analysis of DOX accumulation in MCF7/ADR cells after incubation with CDL for 12 h or 18 h with or without 671 nm laser irradiation (0.25 W·cm⁻², 15 s).  Laser irradiation was executed at 12 h.  n=3, x±s.  ^**P<0.01

 

Part

4

 

细胞毒性

细胞内¹O₂的产生

细胞内ATP水平

细胞周期

Figure 5  A: Cell viabilities of MCF7 (i) and MCF7/ADR (ii) cells after treatment with free DOX, SDL, SCL, CDL or SDL SCL mixture for 48 h at different concentrations. In addition to the special note, the experimental condition of laser group was 2 min, 2 W·cm⁻², 671 nm and the laser irradiation was executed at 12 h.  The mass ratio of DOX to Ce6tM was 5:1; B: Cell viabilities of MCF7/ADR cells after treatment with SDL, SCL and CDL for 48 h at different concentrations with or without laser irradiation (671 nm, 0.25W·cm⁻², 15 s); C: Determination of ¹O₂ in MCF7/ADR cells after incubation with free DOX, SDL, SCL, CDL or SDL SCL mixture for 12 h with or without laser irradiation; D: ATP levels in MCF7/ADR cells after incubation with free DOX, SDL, SCL, CDL, and SDL SCL mixture for 24 h; E: Flow cytometry analysis for cell cycle changes of MCF7/ADR cells induced by free DOX, SCL, SDL, CDL or SDL SCL mixture for 24 h.  n=3, x±s.  ^***P<0.001

Part
5

 

DOX体内分布

Figure 6  Quantitative analysis of in vivo biodistribution of DOX in MCF7/ADR tumor-bearing mice at 4 h and 24 h after intravenous administration of free DOX, SDL, CDL and SDL SCL mixture.  671 nm laser irradiation (2 W·cm⁻², 2 min) was executed at 4 h after administration.  The dose of doxorubicin was 5 mg·kg⁻¹. n=3, x±s.  ^**P<0.01, ^***P<0.001

 

Part
6

 

药效学评价

Figure 7  Inhibitory effect of CDL on tumor growth.  A: The variation profiles of tumor volumes.  The red arrows indicated the time points for drug administrations; B: Tumor weights at the end of the experiment; C: Picture of tumors on day 14 after first drug admini­stration and representative photos of tumor-bearing mice at the end of the experiment.  n=5, x±s.  ^*P<0.05,^**P<0.01

 

Part
7

 

初步的生物安全性评价

Figure 8  Histopathological analysis of tissue sections stained with HE after treatments with saline, free DOX, SCL, SDL, CDL and CDL laser.  The blue circle indicated the injured myocardium tissue area.  The scale bars represented a distance of 100 μm

  

孟庆硕, 张鹏程, 尹琦, 张志文, 于海军, 李亚平. 光敏感型盐酸多柔比星脂质体逆转乳腺癌耐药 [J]. 药学学报, 2017, 52(5): 809-820.

MENG Qing-shuo, ZHANG Peng-cheng, YIN Qi, ZHANG Zhi-wen, YU Hai-jun, LI Ya-ping. Photo-sensitive liposomes loading doxorubicin hydrochloride reverse drug resistance of breast cancer [J]. Acta Pharmaceutica Sinica, 2017, 52(5): 809-820.