Synthesis of protein vesicles for extending time window of ischemic stroke treatment through microcirculatory thrombolysis

Microcirculatory disturbances are a major barrier to extend the time window of ischemic stroke treatment. Herein, we developed a dynamic template method for the synthesis of protein vesicles, by which urokinase-type plasminogen activator (uPA) was crosslinked by poly(ethylene glycol) via pH sensitive host-guest interaction to form a protein gel-shell to encapsulate superoxide dismutase (SOD) for getting cascade biofunctions of microvessel thrombolysis and reperfusion injury inhibition. On a middle cerebral artery occlusion mouse model, the uPA@SOD vesicles administrated beyond intravenous thrombolysis time window (>4.5 h) could reopen the collateral circulation of brain, thus benefiting the followed interventional recanalization of large vessel in the late window (>6 h). These led to rapid reversion of hypoxia condition and more efficient rescue of injured neurons in the ischemic tissue. The synthesis of protein vesicles is highly beneficial for stroke treatment, meanwhile enabling the facile creation of multi-protein therapeutic systems.