Sphingosine kinase (SphK) overexpression is observed in many cancers, including breast, renal and leukaemia, which leads to increased cellular proliferation, survival and growth. SphK inhibition has been an attractive target for anticancer drug development for the past decade, with SphK inhibitors such as PF-543 and opaganib exhibiting clinical antitumour effects. By exploiting both CB5468139 and PF-543 as structural leads, we hereby report on the first quinoline-5,8-dione-based SphK inhibitor using a fragment-based approach. The quinoline-5,8-dione framework was developed to incorporate two defined regions, namely a polar quinoline core, which links to an aryl lipophilic chain. All synthetic molecules were characterized by NMR and HRMS and assayed against SphK 1 and 2, and molecular docking studies were performed. A subset of compounds was screened for anticancer activity. As the binding site of SphK accommodates the lipophilic tail of sphingosine, we initially set out to explore the substitution of the C(7) aryl moiety to attain eight novel C(7) ether-linked quinoline-5,8-diones, which were screened for SphK1 and SphK2 activity with good potency identified. To improve SphK binding, structural fragments were adapted from PF-543 to participate in hydrogen bonding within the binding site of SphK1. A model study was performed to yield novel compounds through activated C(2) formyl intermediates. Two pyrrolidine-based quinoline-5,8-diones were assayed for SphK activity, with revealing an improvement of SphK1 binding efficacy relative to the parent compound and (and its precursor ). Molecular modelling on the pyrrolidine quinoline-5,8-dione construct revealed favourable docking, low binding energies and opportunities for further improvement. Although the screening of anticancer activity was inconclusive, low micromolar dual SphK1/2 inhibition with the quinoline-5,8-dione framework has been identified for the first time, and a plausible new binding mode has been identified.