In this study, a total of six endophytic fungi have been isolated from Vinca rosea (Apocynaceae) leaves growing in Egypt. The isolated fungi were identified morphologically and microscopically up to species to be; Alternaria phragmospora, Aspergillus awamori, Penicillium duclauxii, Penicillium melinii, Nigrospora sphaerica and Mucor ramosissimus. The extracts of the all identified fungi were screened biologically for antileukemic, cytotoxic, antimalarial, antileishmanial, antimicrobial, antioxidant and anti-inflammatory activities as well as for cannabinoid and opioid receptor binding affinities. Out of the six examined fungal extracts four exhibited promising antimalarial activities, four showed modeate antileukemic activities and two of them exhibited cytotoxic activities, three showed antioxidant activities, three exhibited anti-inflammatory activities, and the only one showed an antifungal activity. Alternaria phragmospora was the most active antimalarial agent inhibiting Plasmodium falciparum D6 and W2 clones with IC50 values of 1.9 and 2.1 μg/mL, respectively. Alternaria phragmospora extractwas subjected to liquid-liquid partition using 90% MeOH, hexane then BuOH and H2O. MeOH and BuOH fractions exhibited strong activities against Plasmodium falciparum D6 clone with IC50 values of 1.3 and 28 μg/mL and W2 clone with IC50 values of 1.4 and 28 μg/mL, respectively. These results may be an excellent opportunity to get a new antimalarial drug derived from endophytic fungi.