Two new (1 and 2), together with two known (3 and 4), quinoline alkaloids were isolated from the stems of Nicotiana tabacum. Their structures were elucidated using spectroscopic methods, including extensive H-1, C-13, and 2D NMR techniques. Compounds 1 and 2 were tested for their anti-tobacco mosaic virus (anti-TMV) activity. The results showed that compounds 1 and 2 exhibited potential anti-TMV activity with inhibition rates of 28.2 and 25.8% respectively.