神经酰胺ⅢB磷脂复合物的制备及其评价

doi:10.3969/j.issn.1001-1803.2023.02.004

摘要/Abstract

摘要：

以神经酰胺ⅢB（Ceramide ⅢB，Cer ⅢB）和大豆卵磷脂（Phospholipid，PC）为原料，无水乙醇为反应溶剂，通过蒸发溶剂法制备Cer ⅢB-磷脂复合物（Ceramide phospholipid complex，CerPC）。通过正交实验优化工艺后，利用傅立叶变换红外光谱（FT IR）、差示扫描量热法（DSC）和扫描电镜（SEM）确定了CerPC的形成机制。采用Franz扩散池的Strat-M^®膜进行体外扩散试验，分析渗透性和保留性。采用鸡胚尿囊膜（CAM）试验对刺激性进行了分析。采用COX-2抑制剂筛选试剂盒测定神经酰胺ⅢB磷脂复合物对炎症因子COX-2的抑制率。结果表明，研究中设计的磷脂复合物制备方式安全有效，可以作为Cer ⅢB的载药系统以提高Cer ⅢB的生物利用度并扩大其应用领域。

关键词: 神经酰胺, 磷脂, 结构, 脂溶性, 安全性, 渗透性

Abstract:

Using ceramide ⅢB （Cer ⅢB） and lecithin as raw materials and anhydrous ethanol as reaction solvent, ceramide ⅢB phospholipid complex （CerPC） was prepared by evaporative solvent method. By orthogonal experiment, CerPC was prepared and confirmed according to the following method. Cer ⅢB （20.0 mg/mL） and phospholipids was added in anhydrous ethanol in a molar ratio of 1∶6 and stirred for 1.0 h at 60 ℃. The formation of CerPC was confirmed by using Fourier transform infrared （FT IR） spectroscopy, differential scanning calorimetry （DSC）, and scanning electron microscopy （SEM）. Permeability and retention were analyzed by an in vitro diffusion test in a Franz diffusion cell with a Strat-M^® membrane. Safety and irritation were analyzed by a chick embryo chorioallantois membrane （CAM） assay. The inhibition rate of CerPC on inflammatory factor COX-2 was determined by using a COX-2 inhibitor screening kit. The results show that the preparation method of the phospholipid complex designed in this study is safe and effective, and thus can be used as a drug delivery system for Cer ⅢB to improve its bioavailability and expand its application field.

Key words: ceramide ⅢB, phospholipid, structure, solubility, safety, permeability

中图分类号:

TQ658

宋冠洁,吴文海,张慧荣,吴悦航,易帆,李丽. 神经酰胺ⅢB磷脂复合物的制备及其评价[J]. 日用化学工业（中英文）, 2023, 53(2): 149-156.

Song Guanjie,Wu Wenhai,Zhang Huirong,Wu Yuehang,Yi Fan,Li Li. Preparation and evaluation of ceramide ⅢB phospholipid complex[J]. China Surfactant Detergent & Cosmetics, 2023, 53(2): 149-156.

图/表 12

图1

表1

表2

表3

表4

表5

图2

图3

图4

图5

图6

表6

参考文献 23

[1]	Li Q, Fang H, Dang E, et al. The role of ceramides in skin homeostasis and inflammatory skin diseases[J]. Journal of Dermatological Science, 2019, 97 (1):2-8 doi: 10.1016/j.jdermsci.2019.12.002
[2]	Spada F, Harrison I P, Barnes T M, et al. A daily regimen of a ceramide-dominant moisturizing cream and cleanser restores the skin permeability barrier in adults with moderate eczema: A randomized trial[J]. Dermatologic Therapy, 2021, 34 (4):e14970.
[3]	Dumbuya H, Yan X, Chen Y, et al. Efficacy of ceramide-containing formulations on UV-Induced skin surface barrier alterations[J]. Drugs Dermatol, 2021, 20 (4):s29-s35.
[4]	Draelos Z D, Baalbaki N H. The effect of a ceramide-containing product on stratum corneum lipid levels in dry legs[J]. Journal of Drugs in Dermatology, 2020, 19 (4):372-376. doi: 10.36849/JDD.2020.4796 pmid: 32272513
[5]	Smeden J V, Bouwstra J A. Stratum corneum lipids: their role for the skin barrier function in healthy subjects and atopic dermatitis patients[J]. Curr Probl Dermatol, 2016, 49: 8-26. doi: 10.1159/000441540 pmid: 26844894
[6]	Chen Zhiwei, Zhang Yanan. Effect of moisturizing antipruritic cream on skin barrier function of atopic dermatitis patients and mice model[J]. Chinese Journal of Dermatology and Venereology of Integrated Traditional and Western Medicine, 2020, 19 (4):4.
[7]	Cho W G, Kim K A, Jang S I, et al. Behaviour of nanoemulsions containing Ceramide ⅢB and stratum corneum lipids[J]. Society of Cosmetic Scientists of Korea, 2018 (1).
[8]	Byung Eui Kim, Donald Y M Leung. Significance of skin barrier dysfunction in atopic dermatitis[J]. Allergy, Asthma & Immunology Research, 2018, 10 (3):207-215.
[9]	Andrew K, Rudd, Neal K, et al. Traceless synthesis of ceramides in living cells reveals saturation-dependent apoptotic effects[J]. Proceedings of the National Academy of Sciences of the United States of America, 2018, 115: 201804266.
[10]	Ma Yuhan, Zhao Yan, Sun Yujun, et al. Application of infrared spectroscopy in the detection of fermentation materials for edible and medicinal fungi cosmetics[J]. Science and Technology Information, 2020, 18 (18):5.
[11]	Chen Jia, Li Minghua, Yu Kunzi, et al. Differential scanning calorimetry for identification of pearl powder and shell powder[J]. Journal of Chinese Traditional Medicine, 2015, 40 (8):4.
[12]	Liang Gang, Wang Zhenting, Wang Yongjian, et al. Application and exploration of scanning electron microscope in material specialty practice teaching[J]. Education in Heilongjiang: Theory and Practice, 2019 (3):2.
[13]	Zhou Gang, Fu Xiaoting, Pan Jingru, et al. Comparative study on in vitro Franz diffusion cell determination of tacrolimus ointment transdermal consistency[J]. Journal of Pharmaceutical Analysis, 2020, 40 (12):2126-2133.
[14]	Mohamed, Tahar, Taha, et al. HET-CAM test. Application to shampoos in developing countries[J]. Toxicology in Vitro, 2017, 45 (3):393-396. doi: 10.1016/j.tiv.2017.05.024
[15]	Bi Wei, Shen Shi, Li Qiang, et al. Screening of active parts of Yunnan Baiyao against neovascularization[J]. Chinese Journal of Experimental Formulae, 2011, 17 (22):3.
[16]	Huang Y, Zhang B, Li J, et al. Design, synthesis, biological evaluation and docking study of novel indole-2-amide as anti-inflammatory agents with dual inhibition of COX and 5-LOX[J]. Eur J Med Chem., 2019, 180: 41-50. doi: S0223-5234(19)30625-7 pmid: 31299586
[17]	Ebada H, Nasra M, Elnaggar Y, et al. Novel rhein-phospholipid complex targeting skin diseases: development, in vitro, ex vivo, and in vivo studies[J]. Drug Delivery and Translational Research, 2020, 11 (3):1107-1118. doi: 10.1007/s13346-020-00833-1
[18]	Telange D R, Patil A T, Pethe A M, et al. Formulation and characterization of an apigenin-phospholipid phytosome (APLC) for improved solubility, in vivo bioavailability, and antioxidant potential[J]. European Journal of Pharmaceutical Sciences, 2017, 108: 36-49. doi: S0928-0987(16)30536-X pmid: 27939619
[19]	Liang G, He X, Zhang Y, et al. A dabigatran etexilate phospholipid complex nanoemulsion system for further oral bioavailability by reducing drug-leakage in the gastrointestinal tract[J]. Nanomedicine Nanotechnology Biology & Medicine, 2017, 12 (4):3605-3616.
[20]	Freag M S, Saleh W M, Abdallah O Y. Self-assembled phospholipid-based phytosomal nanocarriers as promising platforms for improving oral bioavailability of the anticancer celastrol[J]. International Journal of Pharmaceutics, 2018, 535 (1/2):18-26. doi: 10.1016/j.ijpharm.2017.10.053
[21]	Chen Ziling, Xu Yingyu, Liu Xiangmei, et al. Evaluation of the stimulation of chorioallantoic membrane vessels in chicken embryo with detergent[J]. Journal of Experimental Animal & Comparative Medicine, 2019, 39 (5):398-401.
[22]	Park C H, Min S Y, Yu H W, et al. Effects of apigenin on RBL-2H3, RAW264.7, and HaCaT cells: anti-allergic, anti-inflammatory, and skin-protective activities[J]. International Journal of Molecular Sciences, 2020, 21 (13):4620. doi: 10.3390/ijms21134620
[23]	Runping S, Li Y, Yue W, et al. Formulation, development, and optimization of a novel octyldodecanol-based nanoemulsion for transdermal delivery of ceramide ⅢB[J]. International Journal of Nanomedicine, 2017, 12: 5203-5221. doi: 10.2147/IJN.S139975 pmid: 28860748

项目	复合时间/h
项目	0.5	1	2	3
复合率/%	86.10±0.44	88.24±0.89	90.34±0.80	90.06±2.38

项目	投料比（n（Cer ⅢB）∶n（PC））
项目	1∶1	1∶2	1∶3	1∶6	1∶8
复合率/%	41.00±4.03	64.20±0.89	88.60±0.89	92.80±1.05	92.60±2.06

项目	质量浓度/（mg/mL）
项目	10	20	25	30	40
复合率/%	81.80±2.95	84.40±1.11	88.60±0.89	64.40±2.59	73.80±3.36

项目	复合温度/℃
项目	30	40	50	60
复合率/%	82.80±2.36	88.60±0.89	81.40±2.08	89.20±1.09

编号	A/h	B/ （mg/mL）	C	D/℃	复合率/%
1	1	10	1∶3	30	65.4%
2	1	20	1∶6	40	89.6%
3	1	25	1∶8	60	93.2%
4	2	10	1∶6	60	85.2%
5	2	20	1∶8	30	92.6%
6	2	25	1∶3	40	88.4%
7	3	10	1∶8	40	76.8%
8	3	20	1∶3	60	93.0%
9	3	25	1∶6	30	90.9%
K₁	2.482 0	2.273 6	2.468 0	2.508 6
K₂	2.682 0	2.772 0	2.656 6	2.547 6
K₃	2.606 2	2.724 6	2.645 6	2.714 0
R	0.200 0	0.498 4	0.188 6	0.205 4