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. 2022 Sep 5;61(41):e202207508. doi: 10.1002/anie.202207508

Figure 4.

Figure 4

Chemo‐cat DOX as a prodrug for ablation of TAMs. A) Representative flow cytometry contour plots of viable macrophages (untreated) as well as doxorubicin‐treated apoptotic (Annexin V+ PI−) and necrotic (Annexin V+ PI+) macrophages. Percentages of cell death in RAW264.7 macrophages after incubation with the indicated drugs (10 μM, 24 h) at 37 °C. Cells were stained with AF647‐Annexin V (25 nM) and PI (3 μM) before analysis by flow cytometry. Data presented as means±SD (n=3). B) Reagents and conditions: a) compound 7, OSu‐PEG4 maleimide, DIPEA, DMSO, r.t., 16 h, 81 %, b) mCCL2‐SH, pH 6.5, r.t., 2 h, 20 %. C) Dose response curves of doxorubicin and compound 8 in RAW264.7 macrophages. Values presented as means±SD (n=3). D,E) Normalized toxicity (relative to doxorubicin alone) in RAW264.7 macrophages (D) or in TAMs and RMs isolated from tumor‐bearing mouse lungs (E) after incubation with doxorubicin, compound 8 and chemo‐cat DOX (all at 1 μM for 48 h). Values presented as means from 2 independent experiments.