Ceruletide and Alpha-1 Antitrypsin as a Novel Combination Therapy for Ischemic Stroke

Ischemic stroke is a primary cause of morbidity and mortality worldwide. Beyond the approved thrombolytic therapies, there is no effective treatment to mitigate its progression. Drug repositioning combinational therapies are becoming promising approaches to identify new uses of existing drugs to synergically target multiple disease-response mechanisms underlying complex pathologies. Here, we used a systems biology-based approach based on artificial intelligence and pattern recognition tools to generate in silico mathematical models mimicking the ischemic stroke pathology. Combinational treatments were acquired by screening these models with more than 5 million two-by-two combinations of drugs. A drug combination (CA) formed by ceruletide and alpha-1 antitrypsin showing a predicted value of neuroprotection of 92% was evaluated for their synergic neuroprotective effects in a mouse pre-clinical stroke model. The administration of both drugs in combination was safe and effective in reducing by 39.42% the infarct volume 24 h after cerebral ischemia. This neuroprotection was not observed when drugs were given individually. Importantly, potential incompatibilities of the drug combination with tPA thrombolysis were discarded in vitro and in vivo by using a mouse thromboembolic stroke model with t-PA-induced reperfusion, revealing an improvement in the forepaw strength 72 h after stroke in CA-treated mice. Finally, we identified the predicted mechanisms of action of ceruletide and alpha-1 antitrypsin and we demonstrated that CA modulates EGFR and ANGPT-1 levels in circulation within the acute phase after stroke. In conclusion, we have identified a promising combinational treatment with neuroprotective effects for the treatment of ischemic stroke.
Neurovascular Research Laboratory acknowledges funding for this project by PI15/00354 and PI18/00804 grants from Instituto Carlos III; the Spanish neurovascular research network INVICTUS PLUS (RD16/0019/0021) also from the Instituto de Salud Carlos III (co-financed by the European Regional Development Fund, FEDER); the program 2017-SGR-1427 from the Generalitat de Catalunya; and the “Advanced Neuroprotection Repurposing Drugs and Nutraceuticals for Stroke in Andalusia: NARDNIA Project,” and COMBINA2 project both funded by the Andalusian Ministry of Health (PE-0527–2019 and PY20-01351). L.R. is supported by a predoctoral fellowship grant (IFI17/00012), from Instituto de Salud Carlos III.