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Preparation, Characterization, and In Vitro Pharmacodynamics and Pharmacokinetics Evaluation of PEGylated Urolithin A Liposomes

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Abstract

Urolithin A (Uro-A), a metabolite of ellagitannins in mammals’ intestinal tract, displays broad biological properties in preclinical models, including anti-oxidant, anti-inflammatory, and anti-tumor effects. However, the clinical application of Uro-A is restricted because of its low aqueous solubility and short elimination half-life. Our purpose was to develop a delivery system to improve the bioavailability and anti-tumor efficacy of Uro-A. To achieve this goal, urolithin A–loaded PEGylated liposomes (Uro-A-PEG-LPs) were prepared for the first time and its physicochemical properties and anti-tumor efficacy in vitro were evaluated. The morphology of Uro-A-PEG-LPs displayed a uniform sphere under transmission electron microscope. The particle size, polydispersity index, zeta potential, and encapsulation efficiency of Uro-A-PEG-LPs were 122.8 ± 7.4 nm, 0.25 ± 0.16, − 25.5 ± 2.3 mV, and 94.6 ± 1.6%, respectively. Moreover, Uro-A-PEG-LPs possessed higher stability and could be stably stored at 4°C for a long time. In vitro release characteristics indicated that Uro-A-PEG-LPs possessed superior sustained release properties. The results of confocal laser scanning microscopy experiment showed that the coumarin 6–loaded PEGylated liposomes (C6-PEG-LPs) have superior cellular uptake than that of conventional liposomes. In addition, in vitro tests demonstrated that Uro-A-PEG-LPs elevated cytotoxicity and pro-apoptotic effect in human hepatoma cells comparing with free Uro-A. Furthermore, the results of pharmacokinetic experiments showed that the t1/2, AUC0-t, and MRT0-t of Uro-A-PEG-LPs increased to 4.58-fold, 2.33-fold, and 2.43-fold than those of free Uro-A solution, respectively. Collectively, these manifested that PEGylated liposomes might be a potential delivery system for Uro-A to prolonging in vivo circulation time, promoting cellular uptake, and enhancing its anti-tumor efficacy.

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Abbreviations

AUC:

Area under the curve

Chol:

Cholesterol

CL:

Plasma clearance

CCK-8:

Cell Counting Kit-8

CLSM:

Confocal laser scanning microscopy

C6:

Coumarin 6

Uro-A:

Urolithin A

Cmax :

Maximum concentration

DSPE-mPEG2000:

1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine (DSPE)–conjugated Polyethylene Glycol 2000

EE:

Encapsulation efficiency

HPLC:

High-performance liquid chromatography

IC50 :

Half-inhibitory concentration

MRT:

Mean residential time

PDI:

Polydispersity index

PEG:

Polyethylene glycol

R:

Range values

SD:

Sprague Dawley

SPC:

Soy phosphatidylcholine

TEM:

Transmission electron microscope

t1/2 :

Half-life time

Uro-A-LPs:

Urolithin A–loaded liposomes

Uro-A-PEG-LPs:

Urolithin A–loaded PEGylated liposomes

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Funding

This study was supported by the National Natural Science Foundation of China (82074077) and the Open Project of Hubei Key Laboratory of Wudang Local Chinese Medicine Research (Hubei University of Medicine) (No. WDCM2019011).

Author information

Author notes

  1. Shengfu Yi and Cong Zhang contributed equally to this work.

Authors and Affiliations

  1. Experimental Training Center, Hubei College of Chinese Medicine, Jingzhou, 434020, People’s Republic of China

    Shengfu Yi

  2. College of Pharmacy, Hubei University of Chinese Medicine, Wuhan, 430065, People’s Republic of China

    Cong Zhang, Junjie Hu, Yan Meng, Liang Chen, Guihong Wang & Zhenpeng Qiu

  3. Hubei Key Laboratory of Wudang Local Chinese Medicine Research, Hubei University of Medicine, Shiyan, 442000, People’s Republic of China

    Huifan Yu & Shan Li

  4. Institute of Biomedicine, Hubei University of Medicine, Shiyan, 442000, People’s Republic of China

    Huifan Yu

  5. Department of Biochemistry, Institute of Basic Medical Sciences, Hubei University of Medicine, Shiyan, 442000, People’s Republic of China

    Shan Li

  6. Key Laboratory for Chinese Medicine Resource and Compound Prescription of Ministry of Education, Hubei University of Chinese Medicine, Wuhan, 430065, People’s Republic of China

    Guohua Zheng

  7. Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan, 430065, People’s Republic of China

    Zhenpeng Qiu

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  1. Shengfu Yi
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  2. Cong Zhang
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  8. Guihong Wang
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Contributions

S.Y. and C.Z.: methodology, investigation, and data curation; C.Z.: writing - original draft; J.H. and Y.M.: software and visualization; L.C.: formal analysis and validation; H.Y. and S.L.: resources. G.H.: supervision. G.Z.: conceptualization and project administration. Z.Q.: funding acquisition and writing - review and editing. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Guohua Zheng or Zhenpeng Qiu.

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Yi, S., Zhang, C., Hu, J. et al. Preparation, Characterization, and In Vitro Pharmacodynamics and Pharmacokinetics Evaluation of PEGylated Urolithin A Liposomes. AAPS PharmSciTech 22, 26 (2021). https://doi.org/10.1208/s12249-020-01890-y

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