Data supporting the article "Modular in vitro evaluation of Buparlisib-polymeric nanomedicines in 2D and 3D models of glioblastoma"

Data set includes the following files: 1. Figures described in the article: Figure 1. Effect of drug modification (SS) on the metabolic activity of GBM cell lines in 2D conditions. Comparison of metabolic activity of 2D-cultured U87MG (A), U118MG (B), and T98G (C) glioblastoma cell lines after a 72-hour treatment with free Buparlisib (Bup; white), pro-drug containing a disulfide bridge (SS-Bup; blue), and polymer-conjugated drug delivery construct (P-SS-Bup; green). Values were normalised to the untreated control group (set to 100 %). Dotted horizontal lines indicate the mean values of the lowest effective free Bup concentration, which were used for the statistical comparison of all groups. Data presented as mean ± SEM. Significance levels (adjusted p - values): [ns] (p_adj ≥ 0.05); [*] (p_adj < 0.05); [**] (p_adj < 0.01); [***] (p_adj < 0.001); [****] (p_adj < 0.0001). n=4‑9 Figure 2. Release kinetics of Buparlisib from the polymer conjugate P-SS-Bup. Buparlisib release from polymer conjugate P-SS-Bup in the presence of 1 mM GSH (25 °C, 75 mM tris‑HCl buffer, pH 7.4). The release is expressed a percentage relative to the complete cleavage with TCEP (100% reference). Figure 3. Effect of drug modification (AP) on the metabolic activity of GBM cell lines in 2D conditions. The effect of free Buparlisib (Bup), Bup derivative containing an azide linker (AP-Bup), and its polymeric conjugated drug delivery construct (P-AP-Bup) on the metabolic activity of 2D-cultivated U87MG (A), U118MG (B), and T98G (C) cells. Values were normalised to the untreated control group (defined as 100 %). Data presented as mean ± SEM. Significance levels (adjusted p - values): [ns] (p_adj ≥ 0.05); [*] (p_adj < 0.05); [**] (p_adj < 0.01); [***] (p_adj < 0.001); [****] (p_adj < 0.0001). n=5‑6 Figure 4. Growth dynamics, metabolic activity, and viability of 3D-cultured U87MG, U118MG, and T98G glioblastoma cell lines. (A) Spheroid size measured over 62 h using the Incucyte® S3 live-cell imaging system. Data presented as means ± SEM. Differences were analysed for each time point. First T98G timepoint value significantly different from both U87MG and U118MG values is depicted as a double circle. Significance levels (adjusted p - values): [**] (p_adj < 0.01); [***] (p_adj < 0.001); [****] (p_adj < 0.0001). n=6‑8 (B) Live/dead staining of 3 days old U87MG, U118MG, and T98G spheroids using fluorescein diacetate (FDA) (live cells, green) and propidium iodide (PI) (dead cells, red); representative images from one biological replica. (C) Metabolic activity of spheroids measured after 3 days of culture. Data presented as mean ± SEM . Significance levels (adjusted p - values): [ns] (p_adj ≥ 0.05); [***] (p_adj < 0.001); [****] (p_adj < 0.0001). n=7 Figure 5. Effect of drug modification (SS) on the metabolic activity of GBM cell lines in 3D conditions. The effect of 72-hour treatment with free Buparlisib (Bup; red), pro-drug containing a disulfide bridge (SS-Bup; blue), and polymer-conjugated drug delivery construct (P-SS-Bup; green) in 50 µM and 200 µM concentrations on metabolic activity of 3D-cultured U87MG (A) and U118MG (B) glioblastoma cell lines. Data presented as mean relative fluorescence units (RFU) per group ± SEM. Significance levels (adjusted p ‑ values): [ns] (p_adj ≥ 0.05); [**] (p_adj < 0.01); [***] (p_adj < 0.001); [****] (p_adj < 0.0001). n=9 Figure 6. Effect of drug modification (SS) on the growth dynamics of GBM cell lines in 3D conditions. The effect of 72-hour treatment with free Buparlisib (Bup; red), pro-drug containing a disulfide bridge (SS-Bup; blue), and polymer-conjugated drug delivery construct (P-SS-Bup; green) in 50 µM and 200 µM concentrations on growth dynamics of 3D-cultured U87MG (A) and U118MG (B) glioblastoma cell lines. Normalised data presented as means (SEM not shown for visual clarity). For each treated group, the first timepoint significantly different from control followed by continuously significant values is depicted as double circle. n=6‑7 2. Supplementary materials. Supplementary Material 1. Supplementary Table 1. Statistical analysis of effectiveness of Bup and its derivatives in comparison to control. Supplementary Material 2. Supplementary Figure 1. Synthesis of AP-Bup derivative scheme.Supplementary Figure 2. Synthesis of SS Bup derivative.scheme.Supplementary Figure3. Synthesis of polymer conjugate P SS Bup scheme.Supplementary Figure 4. Synthesis of polymer conjugate P AP Bup scheme.Supplementary Figure 5. Determination of 100% Buparlisib release using tris(2-carboxyethyl)phosphine (TCEP, 1 mM) under identical conditions (75 mM Tris HCl, pH 7.4, 25 °C). Supplementary Figure 6. Timeline of the treatment regimes for 2D (a) and 3D (b) experiments. Created in BioRender. Havelková, J. (2026) https://BioRender.com/ee01iwb (licensed under CC BY 4.0.). Supplementary Material 3. SEM values omitted in growth dynamics charts (Fig. 6A, 6B). 3. Figures data