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To extend the chemical diversity of jatrophane diterpenoids from Euphorbia peplus and evaluate lysosome-associated phenotypes, phytochemical isolation and a LysoTracker Green assay in HeLa cells were performed. Five previously undescribed jatrophane diterpenoids, Euphjatrophanes M\u2013Q (1\u20135), together with three known analogues (6\u20138), were isolated. The structures of 1\u20135 were assigned from one- and two-dimensional NMR, IR, and UV and the available HR-ESI-MS data, together with a comparison with reported compounds. At 10 and 40 \u03bcM, compounds 1, 2, and 4\u20138 produced higher LysoTracker-associated fluorescence than the DMSO vehicle control, whereas compound 3 showed a small numerical increase at 10 \u03bcM and a decrease at 40 \u03bcM. Compound 1 gave the largest numerical response (1.76-fold and 3.01-fold at 10 and 40 \u03bcM, respectively). 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