In the present study in vitro culture of Leishmania tropica KWH23 (MHOM/PK/2010/KWH23), was used for all the experiments. In axenic growth, Leishmania tropica promastigotes reached to log, mid-log, late-log and stationary phases on day 4, 5, 6 and 7 in culture respectively. Among stationary phase promastigotes higher density of metacyclic was reported. Nectomonad stage promastigotes were found to be the longest and slender most individuals outnumbering any other morphotype for the first three days in the culture. On day 3 onward, leptomonads appeared in the culture to a ratio of 44%. They were distinguished from nectomondas by getting wider anteriorly. Metacyclic promastigotes appeared in culture during log phase with metacyclic to leptomonad to nectomonad ratio of 27, 43 and 29% respectively. During logarithmic growth, 17% of the promastigotes were found to be dividing. Division normally proceeded from flagellum to kinetoplast to nucleus. Amastigote stage was grown in vitro in axenic culture. Day 4 onward most of the parasites in culture were represented by rounded and ovoid cells with no flagella. Cell size decreased from 10.966μm of the promastigote to 3.138μm of round amastigote. During the transformation process 96-98% viability was noted. When the promastigotes were left without fresh medium change, they naturally changed to amastigotes due to pH drop. In a 10 day follow up, the pH dropped from 7.4 to 4.8 and 91% of the parasites, at a density of 1.2x107, changed to amastigotes having 97% viability. These amastigotes were successfully transformed back to promastigotes in normal growth medium. Antileishmanial, cytotoxic and genotoxic effects of Actinomycin D, Z3 and Z5 and Isosteviol derivatives, 16 (2,4-dinitrophenylhydrazine) Isosteviol, 17-hydroxy 16 (2,4- dinitrophenylhydrazine) Isosteviol and Benzyl ester 16 (2,4-dinitrophenylhydrazine) Isosteviol were assessed. Miltefosine was used as standard positive control. Cytotoxicity was expressed i]n terms of 50% inhibitory (IC50) values. The IC50 values of Miltefosine, 16 (2,4- dinitrophenylhydrazine) Isosteviol, 17 hydroxy, 16 (2,4-dinitrophenylhydrazine) Isosteviol, Benzyl ester 16 (2,4-dinitrophenylhydrazine) Isosteviol, actinomycin D, actinomycin Z3 and actinomycin Z5 were 272.2μM (95 % CI= 143.6 to 515.7μM), 781.2μM (95 % CI= 240.8 to 2535.0μM), 294.1μM (95 % CI= 177.4 to 487.5μM), 421.8μM (95% CI= 211.3 to 842.1μM), 195.8μM (95% CI=135.3 to 283.2μM), 210.1μM (95% CI= 145.2 to 304.1μM), and 234.9μM (95% CI= 155.5 to 354.9μM) respectively regarding cytotoxicity. Regarding antileishmanial activity, Miltefosine, 16 (2,4-dinitrophenylhydrazine) Isosteviol, 17- hydroxy 16 (2,4-dinitrophenylhydrazine) Isosteviol, Benzyl ester 16 (2,4- dinitrophenylhydrazine) Isosteviol, Actinomycin D, Actinomycin Z3 and Actinomycin Z5 gave IC50 values against L. tropica promastigotes and amastigotes as 10.80μM (95% CI=9.114 to 12.81μM), 1.245μM (95% CI=0.7250 to 2.138μM), 7.098μM (95% CI=5.328 to 9.455μM), 4.447μM (95% CI= 2.788 to 7.094μM), 5.603μM (95% CI= 4.628 to 6.784μM), 5.033μM (95% CI= 3.189 to 7.945μM), 10.71μM (95% CI= 8.611 to 13.31), 6.794μM (95% CI= 4.248 to 10.87μM), 8.739μM (95% CI= 6.675 to 11.44μM), 2.135μM (95% CI= 1.419 to 3.211μM), 5.500μM (95% CI= 3.811 to 7.939μM), 1.760μM (1.136 to 2.728μM), 9.529μM (95% CI= 7.354 to 12.35μM) and 1.691μM (95% CI= 0.9559 to 2.991μM) respectively. In conclusion, L. tropica KWH23 was extra sensitive to 16 (2,4-dinitrophenylhydrazine) Isosteviol. Miltefosine gave least genotoxicity at 100, 25 and 1.25μM concentration having total comet score (TCS) of 10, 8 and 8 respectively. Damage was non-significant (P>0.05) as compared to 1% DMSO and negative control. Compound 16 (2,4-dinitrophenylhydrazine) Isosteviol showed concentration dependent genotoxicity. It gave TCS values of 207.33, 87.33 and 10.66 respectively at 100, 25 and 4.447μM concentration. The compound showed non-significant genotoxic effects to the standard Miltefosine and 1% DMSO and negative control at all the concentration (P>0.05). Compound 17 hydroxy, 16 (2,4-dinitrophenylhydrazine) Isosteviol caused significant genotoxicity as compared to standard and negative control at 100 and 25μM (P<0.05). At 5.033μM concentraton, however, the genotoxicity became non-significant (P>0.05). Benzyl ester 16 (2,4-dinitrophenylhydrazine) Isosteviol was found to be non-genotoxic as contrasted with the standard and negative control at all concentrations (P>0.05). The TCS values calculated were 166.33, 85.33 and 15 respectively at 100, 25 and 6.794μM concentrations. Actinomycin D showed highest degree of genotoxicity as compared to the standard, Isosteviol derivatives and negative control at 100 and 25μM concentrations (P<0.05). But Genotoxicity became non-significant at 2.135μM concentrations (P>0.05). Actinomycin Z3 was found to show significant genotoxicity at all the concentration i.e., 100, 25 and 1.76μM in relation to standard and negative control as well as Isosteviol derivatives (P<0.05). Actinomycin Z5 was also found significantly genotoxic as compared to standard, negative control and Isosteviol derivatives at all the concentration (P<0.05). In terms of TCS values the genotoxicity, however, greatly reduced with decreasing concentration 100μM (337.666), 25μM (214.333), 1.691μM (23.666).