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Combination of Polyethylene Glycol-Conjugated Urokinase Nanogels and Urokinase for Acute Ischemic Stroke Therapeutic Implications

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Abstract

Tissue reperfusion is a serious therapeutic strategy of ischemic stroke in addition to recanalization. In this work, we aimed to establish new urokinase-based therapeutics in order to dissolve large vessel thrombus together with microthrombi for stroke implications. Formulations consisted of free urokinase (UK), polyethylene glycol–crosslinked urokinase nanogel (PEG-UK), and a 1:1 mixture of UK and PEG-UK (PEG-UK+UK) were tested both in vitro and in vivo. In vitro experiments confirmed the pH-dependent release of PEG-UK in the PEG-UK+UK formulation. It was activated at pH 6.50, an environmental pH in the infarct brain tissue, owing to the dynamic crosslink property of PEG-UK. In vivo tests on a thromboembolic stroke rat model showed that the formulations containing UK, i.e., free UK and PEG-UK+UK, demonstrated better neurological scores and smaller infarction volumes within the time window, in which the PEG-UK+UK formulation relatively performed better. On the other hand, the formulations containing PEG-UK, i.e., PEG-UK and PEG-UK+UK, gained sufficient thrombolytic efficiency beyond the time window. Further investigation on the mechanism revealed that PEG-UK could reduce microthrombus in distal microcirculation, and its destructive effect was also less than that of free UK. The PEG-UK+UK formulation actually provided a “dual targeting” delivery of UK to both the large vessels and the microcirculation, which was beneficial to the treatment of cerebral ischemic stroke both within and beyond the therapeutic time window.

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Abbreviations

CCA:

common carotid artery

ECA:

external carotid artery

Hb:

hemoglobin

H&E:

hematoxylin-eosin

HT:

hemorrhagic transformation

ICA:

internal carotid artery

MCAO:

middle cerebral artery occlusion

MRA:

magnetic resonance angiography

OD:

optical density

OHC-PEG-CHO:

benzaldehyde-terminated PEG

PEG:

polyethylene glycol

PEG-UK:

PEG-urokinase nanogels

SEM:

scanning electron microscopy

TTC:

2,3,5-triphenyltetrazolium chloride

UK:

urokinase

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Acknowledgments

The authors thank Zhang Baogui from the Chinese Academy of Sciences-Institute of Automation Center for Advanced Imaging for the MR scan of rats.

Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 81971115, No. 81400941, No. 51773119, No. 82071306).

Author information

Author notes

  1. Ding Nan and Haiqiang Jin contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, Peking University First Hospital, No. 8 Xishiku Street, Xicheng District, Beijing, 100034, China

    Ding Nan, Haiqiang Jin, Weiwei Yu, Jingjing Jia, Zemou Yu, Yongan Sun, Hongjun Hao & Yining Huang

  2. Center of Materials Science and Optoelectronics Engineering, College of Materials Science and Opto-Electronic Technology, University of Chinese Academy of Sciences, No. 19A, Yuquan Road, Shijingshan District, Beijing, 100049, China

    Di Yang & Xiaozhong Qu

  3. Department of Neurosurgery, Health Science Center, The First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China

    Hui Tan

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  1. Ding Nan
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Contributions

Ding Nan and Haiqiang Jin performed the experiments, analyzed and interpreted the data, and wrote the manuscript; Di Yang, Weiwei Yu, Jingjing Jia, Zemou Yu, Yongan Sun, Hongjun Hao, and Hui Tan aided in the preparation of the manuscript; Xiaozhong Qu supervised the study; Yining Huang supervised the study and provided financial support; and all authors revised the manuscript and approved its final version.

Corresponding authors

Correspondence to Xiaozhong Qu or Yining Huang.

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The authors declare that they have no conflict of interest.

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All experiments were performed according to the Guideline for the Care and Use of Experimental Animals of Peking University First Hospital (J201890).

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First author: Ding Nan and Haiqiang Jin

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Nan, D., Jin, H., Yang, D. et al. Combination of Polyethylene Glycol-Conjugated Urokinase Nanogels and Urokinase for Acute Ischemic Stroke Therapeutic Implications. Transl. Stroke Res. 12, 844–857 (2021). https://doi.org/10.1007/s12975-020-00865-2

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