同轴静电喷涂法制备核壳靶向示踪重组人白细胞介素II微球

doi:10.5246/jcps.2024.06.037

中国药学(英文版) ›› 2024, Vol. 33 ›› Issue (6): 495-510.DOI: 10.5246/jcps.2024.06.037

同轴静电喷涂法制备核壳靶向示踪重组人白细胞介素II微球

朱源¹, 许佳琦¹, 陈小艳¹, 冯颖淑³, Caleb Kesse Firempong¹, 何海冰⁴^,⁵^,^*(), 刘宏飞¹^,²^,⁴^,^*()

^1. 江苏大学药学院, 江苏镇江 212013
^2. 江苏苏南药业工业有限公司, 江苏镇江 212400
^3. 镇江医学院医学化学研究所, 镇江市功能化学重点实验室, 江苏镇江 212028
^4. 江苏海之宏生物医药有限公司, 江苏南通 226001
^5. 沈阳药科大学药学院, 辽宁沈阳 110016

收稿日期:2024-02-06 修回日期:2024-02-25 接受日期:2024-03-29 出版日期:2024-06-30 发布日期:2024-06-30
通讯作者: 何海冰, 刘宏飞

Preparation of core-shell targeted tracer recombinant human interleukin II microspheres via coaxial electrostatic spraying

Yuan Zhu¹, Jiaqi Xu¹, Xiaoyan Chen¹, Yingshu Feng³, Caleb Kesse Firempong¹, Haibing He⁴^,⁵^,^*(), Hongfei Liu¹^,²^,⁴^,^*()

¹ College of Pharmacy, Jiangsu University, Zhenjiang 212013, Jiangsu, China
² Jiangsu Sunan Pharmaceutical Industrial Co., Ltd., Zhenjiang 212400, Jiangsu, China
³ Zhenjiang Key Laboratory of Functional Chemistry, Institute of Medicine & Chemical Engineering, Zhenjiang College, Zhenjiang 212028, Jiangsu, China
⁴ Jiangsu Haizhihong Biomedical Co., Ltd., Nantong 226001, Jiangsu, China
⁵ Department of Pharmaceutics, Shenyang Pharmaceutical University, Shenyang 110016, Liaoning, China

Received:2024-02-06 Revised:2024-02-25 Accepted:2024-03-29 Online:2024-06-30 Published:2024-06-30
Contact: Haibing He, Hongfei Liu
Supported by:
2023 Nantong Jianghai Talent Plan (Grant No. 2023A053), the 2021 Zhenjiang Sixth ‘169 Project’ Scientific Research Project, the 2021 Jurong Social Development Science & Technology Program (Grant No. ZA42109), the 2022 New Drugs and Platform Enhancement Project of the Yangtze Delta Drug Advanced Research Institute, the Zhenjiang Science and Technology Project (Grant No. SH2020048), the China Postdoctoral Science Foundation (Grant No. 2020M681532), the Jiangsu Planned Projects for Postdoctoral Research Funds (Grant No. 2020Z209), and the Natural Science Research Projects of Universities in Jiangsu Province (Grant No. 20KJD350001).

摘要/Abstract

摘要：

重组人白细胞介素-2(rhIL-2)具有促进免疫细胞增殖和分化的潜能, 可用于治疗肺癌。然而, rhIL-2半衰期短, 生物活性不稳定, 需要将其载入微球中进行缓释给药。本研究采用同轴静电喷涂技术制备了载rhIL-2的核壳微球。以量子点为示踪材料, 核层用壳聚糖做载体包裹rhIL-2和量子点偶联物, 壳层采用透明质酸做载体制备核壳复合微球。采用单因素法对载体浓度、电压、针孔内径、喷雾流量等因素进行了研究。优化了同轴静电喷雾法制备核壳微球的工艺参数。结果表明, 制备的核壳微球粒径在1.2–2.0 μm, 包封率和载药量分别为78.39% ± 1.96%和19.58 ± 2.76 μg/mg。体外释放实验显示微球释放效果较好, 无突释现象。该制剂的生物学活性实验表明, 核壳微球中的rhIL-2与游离蛋白药物具有相同的作用。体内成像分析也显示微球具有主动靶向性。本研究为rhIL-2靶向缓释微球的研制提供了理论依据。

关键词: 同轴静电喷雾, 量子点, 透明质酸, 微球

Abstract:

The potential application of recombinant human interleukin-2 (rhIL-2) in promoting immune cell proliferation and differentiation for the treatment of lung cancer is acknowledged. However, the inherent challenges associated with the short half-life and instability of rhIL-2 necessitate its encapsulation into microspheres for sustained release administration. In this study, the coaxial electrostatic spray technique was employed to fabricate rhIL-2-loaded core-shell microspheres. Quantum dots served as tracer materials, and the core-shell composite microspheres were fashioned with chitosan coating the rhIL-2 and quantum dots conjugates in the core layer and hyaluronic acid in the shell layer. A systematic exploration of factors such as carrier concentration, positive voltage, pinhole diameter, and spray flow rate was conducted using a single-factor method. The coaxial electrostatic spray process parameters for core-shell microsphere preparation were also meticulously optimized. The results indicated that the developed core-shell microspheres exhibited a favorable particle size ranging from 1.2 to 2.0 μm, accompanied by encapsulation efficiency and drug loading values of 78.39% ± 1.96% and 19.58 ± 2.76 μg/mg, respectively. In vitro release studies demonstrated a sustained release effect without any discernible burst release phenomenon. Biological activity assessments revealed that rhIL-2 within the core-shell microspheres mirrored the efficacy of the free protein drug. Additionally, in vivo imaging analysis attested to the active targeting properties of the microspheres. These findings robustly supported the successful development of sustained-release targeted rhIL-2-loaded microspheres, providing a theoretical foundation for protein-microsphere formulations.

Key words: Coaxial spray, Quantum dots, Hyaluronic acid, Microspheres

Supporting:

朱源, 许佳琦, 陈小艳, 冯颖淑, Caleb Kesse Firempong, 何海冰, 刘宏飞. 同轴静电喷涂法制备核壳靶向示踪重组人白细胞介素II微球[J]. 中国药学(英文版), 2024, 33(6): 495-510.

Yuan Zhu, Jiaqi Xu, Xiaoyan Chen, Yingshu Feng, Caleb Kesse Firempong, Haibing He, Hongfei Liu. Preparation of core-shell targeted tracer recombinant human interleukin II microspheres via coaxial electrostatic spraying[J]. Journal of Chinese Pharmaceutical Sciences, 2024, 33(6): 495-510.

图/表 16

Figure 1. Effect of different acetic acid concentrations on the particle size and morphology of microspheres (A: 5%; B: 2%; C: 1%).

Figure 2. Effect of different PVA concentrations on the size and morphology of microspheres (A: 5%; B: 4%; C: 3%; D: 2%; E: 1%).

Figure 3. Effect of different CS concentrations on the size and morphology of microspheres (A: 5%; B: 4%; C: 3%; D: 2%; E: 1%).

Figure 4. Effect of different velocities on the particle size of microspheres (A: 0.02 mm/min; B: 0.05 mm/min; C: 0.1 mm/min; D: 0.2 mm/min).

Figure 5. Effect of different voltages on the particle size of microspheres (A: 25 kV; B: 20 kV; C: 15 kV; D: 10 kV).

Figure 6. Effect of different receiving distances on the particle size of microspheres (A: 12 cm; B: 15 cm; C: 18 cm; D: 21 cm).

Figure 7. Effect of different needle diameters on the particle size of microspheres (A: 0.5 mm; B: 0.67 mm; C: 0.86 mm).

Figure 8. Effect of different sodium hyaluronate concentrations on the size and morphology of microspheres (A: 2%; B: 5%; C: 10%; D: 15%).

Figure 9. The effect of different solvent ratio mixtures on the particle size and morphology of microspheres (A: 1:1; B: 1:2; C: 1: 3).

Figure 10. Effect of different receiving distances on the particle size of microspheres (A: 21 cm; B:18 cm; C: 15 cm; D: 12 cm).

Figure 11. Morphological characterization of core-shell microspheres (A: Surface morphology of microspheres; B: Internal structure of microspheres).

Figure 12. Fluorescence stability of core-shell microspheres.

Figure 13. Bioactivity of microspheres with different concentrations.

Figure 14. Characterization of in vitro release of core-shell microspheres.

Table 1. In vitro release curve fitting of microspheres.

Figure 15. In vivo distribution of CS-rhIL-2-QDs-MS(A–C) and HA-CS-rhIL-2-QDs-MS (D–F) after i.v. administration.

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同轴静电喷涂法制备核壳靶向示踪重组人白细胞介素II微球

Preparation of core-shell targeted tracer recombinant human interleukin II microspheres via coaxial electrostatic spraying

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