Viral infections, diabetes, inflammations, and hepatic diseases are worldwide threats nowadays, so effective novel therapeutics are still urgently needed. Fungi always generated significant interest in drug discovery programmer's due to their immense potential to contribute to the discovery of new bioactive compounds. This study was carried out to investigate bioactive compounds from the secondary metabolites of Neosartorya fennelliae. The fractionation of fungal secondary extract to get pure compounds was carried out using Thin Layer Chromatography (TLC). Three active compounds were obtained, chemical characterization of the separated compounds lead to the suggested chemical names as Compound(1) as 4, 5-dihydro-4, 6-bis-(4-methoxyphenyl)-3(2H)-Pyridazinone, compound (4) as1-Hydroxy-2-(2'-methylprop-2'-enyl)-3-(prop-1''-enyl) anthraquinone and compound(10) as 5,7-dihydroxy-6-(3-methyl-2-butenyl)-8-(1-oxobutyl)-4-propyl-2H-1-Benzopyran-2-one. Compound (4) shows promising antiviral, anti-diabetes, and hepatoprotive followed by compound (10), while compound (1) has the best anti-inflammatory activity. So Neosartorya fennelliae considered as a potential source of bioactive compounds.