Intracerebral haemorrhage (ICH) is an unmet medical need leading to the formation of an intracerebral haematoma. Updated guidelines for the management of ICH patients recognize that minimally invasive approaches for evacuation of hematoma open a new area for treatment. Here we demonstrated that O2L-001 allowing the extended-release of a new rtPA variant named OptPA, offers an improved efficacy for haematoma evacuation as well as the best safety.OptPA was produced in CHO cell line before purification, nanoprecipitation using the NANOp2Lysis® technological platform followed by suspension in a solution of 17% poloxamer 407 to obtain O2L-001. Ex vivo hematoma models using human blood were used to show O2L-001 thrombolysis properties and efficacy.Although OptPA has the same thrombolytic activity than rtPA, OptPA has less capacity to activate plasminogen into plasmin in the absence of fibrin, reducing potential bleeding events. We demonstrate that long term exposure to thrombolytic agent were essential to achieve high thrombolysis efficacy. This discovery led to the development of O2L-001 allowing the extended release of OptPA in the first 6h of treatment. Interestingly, unlike rtPA, O2L-001 is able to induce the complete lyse of 5ml hematoma. On clinical size hematomas (obtain from 30 ml of human blood), a single injection of O2L-001 at 1mg/ml into the core of the hematoma led to an increase of thrombolysis of 44% when compared to rtPA.O2L-001 is an interest candidate for ICH treatment and clinical size hematoma is relevant to move toward a First-in-Patient trial.