D-最优混料设计制备青藤碱微乳及其药效学初步考察

高勤; 陈楠; 贾乐彤; 汪洁; 陈静; 杨雨微; 吕志阳

doi:10.14148/j.issn.1672-0482.2024.0391

D-最优混料设计制备青藤碱微乳及其药效学初步考察

doi: 10.14148/j.issn.1672-0482.2024.0391

南京中医药大学翰林学院, 江苏泰州 225300

基金项目:

江苏省“青蓝工程”中青年学术带头人资助项目 2021年

江苏省大学生创新创业训练计划项目 202313981009Y

江苏省大学生创新创业训练计划项目 202313981010Y

详细信息

作者简介:
高勤, 女, 讲师, E-mail:125579431@qq.com

通讯作者:
吕志阳, 男, 副教授, 主要从事药物新型给药系统及适宜性研究, E-mail:lvpharm@163.com

中图分类号: R283
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- 文章访问数: 30
- HTML全文浏览量: 3
- PDF下载量: 2
- 被引次数: 0
出版历程
- 收稿日期: 2023-12-14
- 网络出版日期: 2024-04-24
- 发布日期: 2024-04-10

Preparation of Sinomenine Microemulsion by D-Optimal Mixture Design and Evaluation of Its Pharmacodynamics

Hanlin College, Nanjing University of Chinese Medicine, Taizhou 225300, China

摘要

摘要: 目的优化青藤碱微乳的制备工艺, 进行初步药效学评价。方法建立青藤碱的HPLC含量测定方法, 通过溶解度实验及伪三元相图等初步筛选青藤碱微乳处方, 利用D-最优混料实验设计法以粒径和载药量为考察指标进行青藤碱微乳处方优化, 并对其粒径、载药量及稳定性等进行评价; 采用体外透皮实验考察经皮吸收情况, 进行耳肿胀实验评价抗炎药效。结果以甲醇∶0.1%磷酸(40∶60)为流动相, 检测波长为262 nm测定青藤碱含量, 方法适用性好; 获得微乳最佳处方为蓖麻油占比7.0%, PEG40氢化蓖麻油/无水乙醇为混合乳化剂占比69.0%, 最佳Km值为3∶1, 蒸馏水占比24.0%;制得微乳平均粒径18.76 nm, PDI为0.072, 载药量5.225%;1.0%青藤碱微乳12 h累积渗透量为1.223 4 μg·cm^-2, 稳态渗透速率为0.064 9 μg·cm^-2·h^-1, 均优于青藤碱溶液; 高浓度的青藤碱微乳对小鼠耳肿胀抑制率为65.07%, 与阳性药地塞米松效果相近。结论制得青藤碱微乳工艺稳定, 载药量高, 透皮吸收及抗炎效果较好, 为青藤碱经皮给药制剂开发提供参考。
- 青藤碱 /
- 微乳 /
- 伪三元相图 /
- D-最优混料设计 /
- 药效学
Abstract: OBJECTIVE To optimize the preparation process of sinomenine microemulsion and evaluate its pharmacodynamics. METHODS HPLC method for sinomenine content determination was established, and sinomenine microemulsion prescription was initially screened by solubility test and pseudo-ternary phase diagram. D-optimal mixing experimental design method was used to optimize sinomenine microemulsion prescription with particle size and drug load as investigation indexes, and its particle size, drug load and stability were evaluated. Transdermal absorption was investigated by transdermal test in vitro, and the anti-inflammatory effect was evaluated by ear swelling test. RESULTS With methanol:0.1% phosphoric acid (40∶60) as the mobile phase, the detection wavelength was 262 nm, and the method was suitable for the determination of sinomenine. The optimal formula of microemulsion was obtained as castor oil (7.0%), PEG40 hydrogenated castor oil/anhydrous ethanol (69.0%), the optimal Km value was 3∶1, and distilled water (24.0%). The average particle size of the microemulsion was 18.76 nm, the PDI was 0.072 and the drug loading was 5.225%. The cumulative permeability of 1.0% sinomenine microemulsion at 12 h was 1.223 4 μg·cm^-2, and the steady permeability rate was 0.0649 μg·cm^-2·h^-1, which was better than sinomenine solution. The inhibitory rate of sinomenine microemulsion was 65.07%, which was similar to dexamethasone. CONCLUSION The preparation of sinomenine microemulsion has the advantages of stable process, high drug loading, good transdermal absorption and anti-inflammatory effect, which can provide reference for the development of sinomenine transdermal drug delivery preparation.
- sinomenine /
- microemulsion /
- pseudo-ternary phase diagram /
- d-optimal mixture design /
- pharmacodynamics

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图 1 伪三元相图

Figure 1. The pseudo-ternary phase diagrams

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图 2 X₁和X₂添加量配比对综合评分的影响

Figure 2. The effect of the ratio of X₁ and X₂ on the comprehensive score

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图 3 X₁和X₃添加量配比对综合评分的影响

Figure 3. The effect of the ratio of X₁ and X₃ on the comprehensive score

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图 4 X₂和X₃添加量配比对综合评分的影响

Figure 4. The effect of the ratio of X₂ and X₃ on the comprehensive score

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图 5 青藤碱微乳的粒径分布图

Figure 5. The particle size distribution of sinomenine microemulsion

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图 6 不同体系青藤碱的渗透动力学曲线

Figure 6. Osmotic kinetic curves of sinomenine in different systems

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表 1 青藤碱在各辅料中溶解度

Table 1. Solubility of sinomenine in excipients

微乳组成	种类	溶解度/(mg·g^-1)
油相	蓖麻油	0.450 4
	油酸乙酯	0.003 8
	油酸	0.059 7
乳化剂	聚山梨酯80	1.109 7
	聚山梨酯20	1.075 7
	PEG40氢化蓖麻油	1.815 4
助乳化剂	无水乙醇	3.477 5
助乳化剂	1, 2-丙二醇	2.928 7

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表 2 乳化等级分类

Table 2. Emulsification grade classification

乳化等级	外观描述
A	快速乳化, 外观呈澄清或淡蓝色乳光
B	快速乳化, 外观呈半透明或蓝白色
C	较快乳化, 外观呈亮白色不透明液体
D	乳化较慢, 外观呈暗灰色或上部略带油滴
E	不发生乳化, 上部有油滴

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表 3 青藤碱微乳辅料配伍实验结果

Table 3. Experimental results of sinomenine microemulsion excipient compatibility

油相	乳化剂	助乳化剂	现象	放置 3d 后现象
油酸	聚山梨酯80	无水乙醇	C	C
蓖麻油	聚山梨酯80	无水乙醇	E	E
油酸	聚山梨酯80	1, 2-丙二醇	C~D	D
蓖麻油	聚山梨酯80	1, 2-丙二醇	D~E	D-E
油酸	PEG40氢化蓖麻油	无水乙醇	A~B	B
蓖麻油	PEG40氢化蓖麻油	无水乙醇	A	A
油酸	PEG40氢化蓖麻油	1, 2-丙二醇	B	B~C
蓖麻油	PEG40氢化蓖麻油	1, 2-丙二醇	B~C	A~B

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表 4 D-最优混料设计及结果(x±s,n=3)

Table 4. D-optimal mixture design and results(x±s,n=3)

序号	X₁/%	X₂/%	X₃/%	Y/nm	Z/%	综合评分M
1	28.5	31.5	40.0	115.25	6.128	140
2	43.3	26.1	30.6	137.50	4.007	100
3	7.0	69.0	24.0	18.86	5.225	170
4	69.5	10.0	20.5	180.73	0.040	30
5	62.8	26.1	11.1	192.03	0.143	20
6	7.0	53.0	40.0	125.25	4.459	100
7	89.0	10.0	1.0	122.83	0.114	50
8	7.0	61.0	32.0	28.24	3.460	140
9	21.8	47.6	30.6	78.13	1.418	90
10	50.0	10.0	40.0	95.38	7.446	160
11	36.6	41.2	21.2	115.10	2.350	90
12	30.0	69.0	1.0	54.06	3.056	130
13	21.8	55.6	22.6	38.28	5.241	160
14	18.5	69.0	12.5	40.55	2.510	130
15	33.3	55.6	11.1	56.95	4.403	140
16	59.5	39.5	1.0	147.90	0.591	40

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表 5 青藤碱各体系体外透皮吸收的比较(n=3)

Table 5. Comparison of transdermal absorption of sinomenine systems in vitro (n=3)

体系	回归方程	R²	Q₁₂/(μg·cm^-2)	J_s/(μg·cm^-2·h^-1)
1%青藤碱水溶液	Q=0.046 0t+0.4516	0.983 2	1.003 1±0.004 8	0.046 0
1%青藤碱微乳	Q=0.064 9t+0.450 9	0.995 5	1.223 4±0.019 5	0.064 9

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表 6 青藤碱微乳对二甲苯所致小鼠耳肿胀的影响(x±s,n=8)

Table 6. Effect of sinomenine microemulsion on xylene induced ear swelling in mice(x±s,n=8)

组别	剂量/(g·kg^-1)	肿胀度/mg	肿胀率/%	抑制率/%
空白组	—	0	0	—
模型组	—	43.8±1.8	99.54	—
地塞米松组	0.005	16.6±1.3^**	25.70	74.18
低浓度微乳组	0.500	40.0±2.6^**	85.65	13.95
高浓度微乳组	2.000	24.2±1.1^**	34.77	65.07
注: 与模型组比较，^P < 0.05, ^*P < 0.01。

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