Glioblastoma multiforme (GBM) is the most common and highly aggressive form of brain cancer. Frontline therapy involves surgical resection followed by radiotherapy combined with the chemotherapeutic alkylating agent Temozolomide (TMZ). Despite this multi-modal therapy, many GBM tumours are intrinsically resistant to TMZ making it challenging to treat. The aim of this study is to evaluate the therapeutic potential of co-targeting pro-survival BCL-2 family proteins with BH3-mimetic drugs and the dihydroorotate dehydrogenase (DHODH) inhibitor, BAY-2402234. For in vitro evaluation, we used patient derived glioma neurospheres (PDNs), patient derived brain organoids (PDBOs), flow-cytometry based cell death assays, Western blotting, Incucyte live cell imaging and Cell-Titre glo assays. For in vivo studies, we used healthy immunodeficient NSG mice and immune competent C57BL/6 mice. We demonstrate that co-targeting the pro-survival proteins BCL-XL with ABT-1331852 or MCL-1 with S63845 or treatment with BAY-2402234 combined with A1331852 or S63845 was more potent at killing PDNs compared to monotherapy treatment and conventional therapy with TMZ. The combined loss of the essential effectors of apoptosis, BAX and BAK, rendered PDNs refractory to killing induced by BH3-mimetics. However, these PDNs were still sensitive to treatments with BAY-2402234 plus BH3-mimetics, suggesting that other modes(s) of cell death are also engaged. Co-treatment with A1331852 and S63845 promoted a significant loss in cell viability, spheroid area and invasiveness of PDBOs compared to TMZ. PDBOs co-treated with BAY-2402234 and A1331852 or S63845 displayed a reduction in spheroid area, invasiveness and cell viability compared to vehicle treatment. NSG and C57BL/6 mice tolerated treatment with BAY-2402234 for 10 days, with no adverse alterations in neuronal structure as assessed by cresyl violet staining of brain slices and body weight of mice compared to vehicle control treated cohorts. NSG mice treated with BAY-2402234 exhibited a significant drop in platelet counts 2 days post-drug treatment, however, platelet counts increased within 5 days post-drug treatment and were restored to normal levels after 13 days. These findings demonstrate the potential of dual targeting of pro-survival BCL-2 family proteins and DHODH in GBM. Identifying an ideal therapeutic window may guide the utility of BH3-mimetics and BAY-2402234 for improved therapies for GBM.