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. 2022 Feb 2;3(2):100502. doi: 10.1016/j.xcrm.2021.100502

Figure 7.

Figure 7

Testing the efficacy of SU086 in combination with enzalutamide and abiraterone in vitro and in vivo

(A) Coefficient of drug interaction (CDI) was tested with colony-formation assay of each compound at varied single dose concentrations and combination concentrations - SU086 (250 nM to 1 μM), enzalutamide (ENZ 1 μM to 5 μM), abiraterone (ABI 500 nM to 2.5 μM). CDI = AB (SU086 + ENZ treatment, or SU086 + ABI treatment)/ [(A, SU086 treatment alone) × (B, ENZ or ABI treatment alone)]. Values less than one indicate synergism, values equal to one indicate additive effects, and values greater than one indicate antagonism. Final concentrations were calculated with the final dosing as noted, SU086 (250 nM), ENZ (5 μM), ABI (2.5 μM). As such, SU086 synergizes with ENZ and has additive effects with ABI.

(B) Colony formation of C4-2 cells with SU086 (250 nM), ENZ (5 μM), ABI (2.5 μM), SU086 + ENZ, SU086 + ABI, or DMSO control. Representative images of wells are shown; scale bar, 100 mm.

(C) Tumoroid growth of LuCaP 136 cells embedded in 50% Matrigel in the presence of SU086 (1 μM) plus ENZ (5 μM) or ABI (2.5 μM) for 15 days with medium and compounds changed every other day. Wells were scanned with Celigo Imager and whole wells counted manually from images in triplicate. Scale bar (top), 300 μm; enlarged, 50 μm. In vitro assays were performed in triplicate with representative images shown, graphed ±SD.

(D) C4-2 xenografts in mice treated with vehicle control, SU086 (50 mg/kg i.p. daily), ENZ (10 mg/kg daily gavage), ABI (200 mg/kg daily gavage), SU086 plus ENZ, or SU086 plus ABI. Treatments were initiated when the average tumor volumes reached 50 mm3. Mice were treated for 21 days with tumors measured every third day. Animals were sacrificed when the average tumor volumes of the vehicle-treated group reached 400 mm3. Tumors were graphed as fold change over day 0 per each individual tumor ± SEM.

(E) H&E and Ki67 IHC of tumors from combination therapy experiments. The Ki67 proliferative index was quantified by manually counting percentage of positive nuclei per image field, averaged for three separate fields and graphed ± SD. Scale bars, 50 μm; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.005.