Ohioensins F and G (1 and 2), two new benzonaphthoxanthenones, have been isolated from the MeOH extract of Antarctic moss Polytrichastrum alpinum by various chromatographic methods. The structures of these compounds were determined mainly by analysis of NMR spectroscopic data. The known compounds ohioensins A and C (3 and 4) were also obtained. Compounds 1-4 showed potent inhibitory activity against therapeutically targeted protein tyrosine phosphatase 1B (PTP1B). Kinetic analysis of PTP1B inhibition by ohioensin F (1) suggested that benzonaphthoxanthenones inhibited PTP1B activity in a non-competitive manner.