Effects of Probucol Formulations on Mesenteric Lymphatic Trans-port Efficiency and Pharmacokinetics in Rats

doi:10.12300/j.issn.1674-5817.2022.026

Abstract

Abstract:

Objective To compare the effects of probucol olive oil and suspension formulations on the pharmacokinetics and mesenteric lymphatic transport in rats using an innovative mesenteric-jugular lymphatic duct assisted reflux model. Methods Twelve Sprague-Dawley (SD) rats were randomly divided into four groups: suspension preparation-jugular vein single cannulation group (H-JD group), suspension preparation-jugular vein and intestinal lymphatic double cannulation group (H-JCS group), olive oil preparation-jugular vein single cannulation group (G-JD group), olive oil preparation-jugular vein and intestinal lymphatic double cannulation group (G-JCS group). The concentrations of probucol in whole blood and lymph fluid of rats at different times were determined by liquid chromatography and tandem mass spectrometry (LC-MS/MS), and drug-time curves were drawn. The pharmacokinetic parameters, relativebioavailability (F_rel) and percentage dose in lymph fluid were calculated. Results The drug-time curve of each group conformed to the non-compartmental model. The peak times (T_max) of the H-JD group, H-JCS group, G-JD group, and G-JCS group were (11±12), (5±2), (13±9), and (19±9) h, respectively; the peak concentrations (C_max) were (148±60), (207±137), (453±204), and (309±177) ng/mL, respectively; the areas under the curve (AUC_last) were (3 210±885), (3 677±2 014), (12 360±6 629), and (8 080±3 064) h·ng·mL^-1, respectively. The percentages of lymphatic fluid dose in the H-JCS and G-JCS groups were (1.29±0.50)% and (2.59±0.43)%. Compared with the H-JD group, the F_rel of probucol in the G-JD group was (409±269)%; compared with the H-JCS group, the F_rel of probucol in the G-JCS group was (309±256)%. Compared with the H-JD and H-JCS groups, the whole blood values of C_max, T_max, AUC_last and percentage of lymphatic fluid probucol concentrations of the G-JD and the G-JCS groups were significantly increased (P＜0.05). The AUC_last of whole blood in the G-JD group was significantly higher than that in the G-JCS group (P＜0.05), but there was no significant difference between the H-JD and H-JCS groups (P > 0.05). Conclusion Olive oil formulation can improve the ratio of probucol transport through mesenteric lymph as well as its bioavailability.

Key words: Probucol, Mesenteric lymph fluid, Pharmacokinetics, Transport, Sprague-Dawley rats

CLC Number:

Q95-33

Xiaorui ZHANG, Jing CAO, Qianqian WU, Jijun LIU, Guoyuan CHEN, Baojin WU. Effects of Probucol Formulations on Mesenteric Lymphatic Trans-port Efficiency and Pharmacokinetics in Rats[J]. Laboratory Animal and Comparative Medicine, 2022, 42(4): 275-283.

Figures/Tables 7

Figure 1 Chromatogram of blood and lymph with and without probucolNote: (A) Blank whole blood; (B) Blank lymph fluid; (C) Probucol (1 ng/mL) added to whole blood; (D) Probucol (1 ng/mL) added to lymph fluid.

Figure 2 Standard curves of probucol in blood and lymphNote：(A) Probucol in whole blood; (B) Probucol in lymph fluid.

Table 1 Drug concentrations in the blood of rats in each group

采血时间 Blood collection time/h	普罗布考Probucol ρ/(ng·mL^-1)
采血时间 Blood collection time/h	H-JD组 H-JD group	H-JCS组 H-JCS group	G-JD组 G-JD group	G-JCS组 G-JCS group
0	-	-	-	-
0.25	-	-	-	-
0.5	5	-	-	-
1	34±7	28±32	13±5	-
2	62±48	38±32	76±30	19±23
4	126±91	188±167	143±26	82±119
8	104±87	100±36	379±138	128±184
24	67±30	69±49	289±296	252±189
48	12±5	22±15	85±23	48±13
72	5±1	11±10	15±5	18±1

Figure 3 Drug concentration-time curve (A) and dose percentage curve of lymphatic fluid (B) in all groups of ratsNote: H-JD, suspension preparation-jugular vein single cannulation group; H-JCS, suspension preparation-jugular vein and intestinal lymphatic double cannulation group; G-JD, olive oil preparation-jugular vein single cannulation group; G-JCS, olive oil preparation-jugular vein and intestinal lymphatic double cannulation group.

Table 2 Pharmacokinetic parameters of the whole blood of rats in each group

参数 Parameters	H-JD组 H-JD group	H-JCS组 H-JCS group	G-JD组 G-JD group	G-JCS组 G-JCS group
峰值时间T_max/h	11±12	5±2	13±9	19±9
峰值浓度C_max/(ng·mL^-1)	148±60	207±137	453±204	309±177
半衰期T_1/2/h	12±1	17±3	13±3	19±11
药-时曲线下面积AUC_last/(h·ng·mL^-1)	3 210±885	3 677±2 014	12 360±6 629	8 080±3 064
药-时曲线下面积AUC_INF/(h·ng·mL^-1)	3 293±899	3 970±2 295	12 623±6 650	8 947±2 218

Table 3 Relative bioavailability (Frel) of rats in each group

动物 Rat	药-时曲线下面积AUC_last/(h·ng·mL^-1)		F_rel/%	药-时曲线下面积AUC_last/(h·ng·mL^-1)		F_rel/%
动物 Rat	H-JD组 H-JD group	G-JD组 G-JD group	F_rel/%	H-JCS组 H-JCS group	G-JCS组 G-JCS group	F_rel/%
第1只No.1	2 750	19 800	720	3 360	8 680	258
第2只No.2	4 230	10 200	241	1 840	10 800	587
第3只No.3	2 650	7 080	267	5 830	4 760	82
平均数 $x ¯$ ±s	3 210±885	12 360±6 629	409±269	3 677±2 014	8 080±3 064	309±256

Table 3 Relative bioavailability (Frel) of rats in each group

动物 Rat	药-时曲线下面积AUC_last/(h·ng·mL^-1)		F_rel/%	药-时曲线下面积AUC_last/(h·ng·mL^-1)		F_rel/%
动物 Rat	H-JD组 H-JD group	G-JD组 G-JD group	F_rel/%	H-JCS组 H-JCS group	G-JCS组 G-JCS group	F_rel/%
第1只No.1	2 750	19 800	720	3 360	8 680	258
第2只No.2	4 230	10 200	241	1 840	10 800	587
第3只No.3	2 650	7 080	267	5 830	4 760	82
平均数 $x ¯$ ±s	3 210±885	12 360±6 629	409±269	3 677±2 014	8 080±3 064	309±256

Table 4 Dose percentage parameters of lymphatic fluid of rats in each group

参数 Parameters	采液时间/h Liquid collection time/h	H-JCS组 H-JCS group	G-JCS组 G-JCS group
质量浓度ρ/（ng·mL^-1）	0~4	2 880±2 120	2 862±968
	4~8	5 682±4 935	4 019±4 019
	8~24	417±189	1 345±1 695
体积V/mL	0~4	6.8±0.5	11.0±2.0
	4~8	7.2±0.6	19.2±1.4
	8~24	20.8±2.4	30.4±3.0
剂量百分比Dose percentage/%	0~4	0.37±0.26	0.60±0.25
	0~8	1.13±0.87	1.85±0.88
	0~24	1.29±0.50	2.59±0.43

References 23

1	HIRATA K I. New evidence of probucol on cardiovascular events[J]. J Atheroscler Thromb, 2021, 28(2):97-99. DOI:10.5551/jat.ED155 .
2	YAMASHITA S, MASUDA D, OHAMA T, et al. Rationale and design of the PROSPECTIVE trial: probucol trial for secondary prevention of atherosclerotic events in patients with prior coronary heart disease[J]. J Atheroscler Thromb, 2016, 23(6):746-756. DOI:10.5551/jat.32813 .
3	LIU J C, LI M H, LU H, et al. Effects of probucol on restenosis after percutaneous coronary intervention: a systematic review and meta-analysis[J]. PLoS One, 2015, 10(4): e0124021. DOI:10.1371/journal.pone.0124021 .
4	HIGASHI K, MORI A, SAKAMOTO K, et al. Probucol slows the progression of cataracts in streptozotocin-induced hyperglycemic rats[J]. Pharmacology, 2019, 103(3-4):212-219. DOI:10.1159/000496055 .
5	JUNG Y S, PARK J H, KIM H, et al. Probucol inhibits LPS-induced microglia activation and ameliorates brain ischemic injury in normal and hyperlipidemic mice[J]. Acta Pharmacol Sin, 2016, 37(8):1031-1044. DOI:10.1038/aps.2016.51 .
6	HAN L M, YANG Q S, SHEN T, et al. Lymphatic transport of orally administered probucol-loaded mPEG-DSPE micelles[J]. Drug Deliv, 2016, 23(6):1955-1961. DOI:10.3109/10717544.2015.1028600 .
7	QIAN Y W, CHEN G G, WANG J, et al. Preparation and evaluation of probucol-phospholipid complex with enhanced bioavailability and No food effect[J]. AAPS Pharmscitech, 2018, 19(8):3599-3608. DOI:10.1208/s12249-018-1157-2 .
8	GERSHKOVICH P, HOFFMAN A. Uptake of lipophilic drugs by plasma derived isolated chylomicrons: linear correlation with intestinal lymphatic bioavailability[J]. Eur J Pharm Sci, 2005, 26(5):394-404. DOI:10.1016/j.ejps.2005.07.011 .
9	YÁÑEZ J A, WANG S W J, KNEMEYER I W, et al. Intestinal lymphatic transport for drug delivery[J]. Adv Drug Deliv Rev, 2011, 63(10-11):923-942. DOI:10.1016/j.addr.2011.05.019 .
10	BEG S, ALAM M N, AHMAD F J, et al. Chylomicron mimicking nanocolloidal carriers of rosuvastatin calcium for lymphatic drug targeting and management of hyperlipidemia[J]. Colloids Surf B Biointerfaces, 2019, 177:541-549. DOI:10.1016/j.colsurfb.2019.02.039 .
11	HOKKANEN K, TIRRONEN A, YLÄ-HERTTUALA S. Intestinal lymphatic vessels and their role in chylomicron absorption and lipid homeostasis[J]. Curr Opin Lipidol, 2019, 30(5):370-376. DOI:10.1097/MOL.0000000000000626 .
12	KIM H, SEONG I, RO J, et al. Enhanced association of probucol with chylomicron by pharmaceutical excipients: an in vitro study[J]. Drug Dev Ind Pharm, 2015, 41(7):1073-1079. DOI:10.3109/03639045.2014.927479 .
13	张笑瑞, 曹静, 吴倩倩, 等. 大鼠肠系膜淋巴液持续引流新方法的初步建立[J]. 实验动物与比较医学, 2022, 42(4):267-274. DOI:10.12300/j.issn.1674-5817.2022.024 .
	ZHANG X R, CAO J, WU Q Q, et al. A preliminary method for continuous drainage of mesenteric lymph fluid in rat[J]. Laboratory Animal and Comparative Medicine, 2022, 42(4):267-274. DOI:10.12300/j.issn.1674-5817.2022.024 .
14	ALEXANDER J S, GANTA V C, JORDAN P A, et al. Gastrointestinal lymphatics in health and disease[J]. Pathophysiology, 2010, 17(4):315-335. DOI:10.1016/j.pathophys. 2009.09.003 .
15	HYEONGMIN K, YEONGSEOK K, JAEHWI L E. Liposomal formulations for enhanced lymphatic drug delivery[J]. Asian J Pharm Sci, 2013(2):96-103.
16	MARWAHA R K, DABAS A. Bioavailability of nanoemulsion formulations vs conventional fat soluble preparations of cholecalciferol (D3) - An overview[J]. J Clin Orthop Trauma, 2019, 10(6):1094-1096. DOI:10.1016/j.jcot.2019.07.014 .
17	ZHANG Y M, ZHANG S K, CUI N Q. Intravenous infusion of mesenteric lymph from severe intraperitoneal infection rats causes lung injury in healthy rats[J]. World J Gastroenterol, 2014, 20(16):4771-4777. DOI:10.3748/wjg.v20.i16.4771 .
18	CHAMPAGNE D, PEARSON D, DEA D, et al. The cholesterol-lowering drug Probucol increases apolipoprotein E production in the hippocampus of aged rats: implications for Alzheimer's disease[J]. Neuroscience, 2003, 121(1):99-110. DOI:10.1016/S0306-4522(03)00361-0 .
19	BROCKS D R, DAVIES N M. Lymphatic drug absorption via the enterocytes: pharmacokinetic simulation, modeling, and considerations for optimal drug development[J]. J Pharm Pharm Sci, 2018, 21(1s):254s-270s. DOI:10.18433/jpps30217 .
20	LI J, YANG Y, ZHAO M H, et al. Improved oral bioavailability of probucol by dry media-milling[J]. Mater Sci Eng C, 2017, 78:780-786. DOI:10.1016/j.msec.2017.04.141 .
21	SCHÖNFELD P, WOJTCZAK L. Short- and medium-chain fatty acids in energy metabolism: the cellular perspective[J]. J Lipid Res, 2016, 57(6):943-954. DOI:10.1194/jlr.R067629 .
22	TANOUS D, BRÄSEN J H, CHOY K, et al. Probucol inhibits in-stent thrombosis and neointimal hyperplasia by promoting re-endothelialization[J]. Atherosclerosis, 2006, 189(2):342-349. DOI:10.1016/j.atherosclerosis.2006.01.025 .
23	KOSTIK V, MEMETI S, BAUER B. Fatty acid composition of edible oils and fats[J]. J Hyg Eng Des, 2013, 4: 112-116.

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