HIGH-THROUGHPUT SCREENING OF BLOOD–BRAIN BARRIER-PENETRANT COMPOUNDS IDENTIFIES THERAPEUTIC CANDIDATES FOR CENTRAL NERVOUS SYSTEM-TROPIC PEDIATRIC ACUTE LYMPHOBLASTIC LEUKEMIA
5 affiliations
- 1Department of Genetics, Ribeirão Preto Medical School, University of São Paulo Ribeirão Preto, SP, Brazil
- 2Boldrini Children's Center, Campinas, SP, Brazil
- 3Department of Pediatrics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, SP, Brazil
- 4Department of Translational Medicine, School of Medical Sciences, State University of Campinas, Campinas, SP, Brazil
- 5Department of Clinical, Toxicological and Food Analysis, Ribeirão Preto School of Pharmaceutical Sciences, University of São Paulo, Ribeirão Preto, SP, Brazil
Abstract
Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy, accounting for approximately 80% of childhood leukemia cases. Despite therapeutic advances, central nervous system (CNS) involvement remains a major clinical challenge, associated with chemoresistance and relapse, partly due to the limited ability of conventional chemotherapeutic agents to cross the blood–brain barrier (BBB). To identify drug repurposing candidates for pediatric ALL, 270 compounds from the CNS-Penetrant Compound Library were screened using the Ex vivo Leukemia Drug Advisor (ELDA) platform, which combines automated real-time imaging and computational analysis. Cells were continuously monitored for 48 hours, enabling the generation of chemosensitivity curves according to drug concentration and exposure time. All evaluated compounds, including library compounds and the positive controls methotrexate, vincristine, and cytarabine, were tested at a fixed concentration of 27 µM. Assay quality was validated by Z-factor analysis, with values ≥0.7 across all evaluated cell lines (Jurkat, Nalm6, RS4;11, SUP-B15, and K562), confirming the robustness and reproducibility of the platform. Hits were identified based on compounds inducing ≥50% reduction in cell viability, followed by integrated analysis to select candidates showing consistent activity across all five cell lines at both 24 and 48 hours of treatment. This strategy identified 16 candidate compounds. Among these, BMS-202, BAY 11-7082, and WP1066 were prioritized based on their translational potential, BBB-penetrating properties, and limited evidence regarding their therapeutic application in pediatric ALL. These compounds were selected for further pharmacological evaluation, including dose–response experiments to determine IC₅₀ values in leukemic cell lines, which are currently underway. To evaluate cytotoxicity in non-malignant cells, two bone marrow-derived cell lines, MSC-hTERT and HS-5, were included. The IC₅₀ values obtained to date after 48 hours of treatment were 37.0 μM and 14.2 μM for BMS-202, 13.5 μM and 7.2 μM for BAY 11-7082, and 15.9 μM and 10.0 μM for WP1066 in MSC-hTERT and HS-5 cells, respectively. Collectively, these findings demonstrate the applicability of a BBB-penetrant compound-based screening strategy for identifying drug repurposing candidates in pediatric ALL. The identification of BMS-202, BAY 11-7082, and WP1066 highlights the potential for new therapeutic applications of molecules that remain poorly explored in this context, expanding treatment opportunities for a disease with unmet clinical needs. These results reinforce the relevance of investigating BBB-penetrant compounds and support further pharmacological and functional characterization of these candidates, particularly for therapeutic strategies targeting patients with CNS involvement. Funding: National Council for Scientific and Technological Development (CNPq), Process No. 168989/2023-9; São Paulo Research Foundation (FAPESP), Process No. 2022/09037-3; Coordination for the Improvement of Higher Education Personnel (CAPES), Finance Code 001; Graduate Program of the Department of Genetics, Ribeirão Preto Medical School, University of São Paulo; Teaching, Research and Assistance Support Foundation of the Hospital das Clínicas, Ribeirão Preto Medical School, University of São Paulo (FAEPA).
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BibTeX
@article{franca2026highthroughput,
title = {{HIGH-THROUGHPUT SCREENING OF BLOOD–BRAIN BARRIER-PENETRANT COMPOUNDS IDENTIFIES THERAPEUTIC CANDIDATES FOR CENTRAL NERVOUS SYSTEM-TROPIC PEDIATRIC ACUTE LYMPHOBLASTIC LEUKEMIA}},
author = {Manuela Eduarda FRANÇA and Jessica Oliveira DE SANTIS and Mariane Minussi BAPTISTELLA and Leandro Henrique DE PAULA ASSIS and Veridiana Kiill SUAZO and Rosane Gomes DE PAULA QUEIROZ and José Andrés YUNES and Graziella Ribeiro SOUSA and Carlos Alberto SCRIDELI},
journal = {Hematology, Transfusion and Cell Therapy},
year = {2026},
month = oct,
doi = {10.1016/j.htct.2026.107598},
url = {https://doi.org/10.1016/j.htct.2026.107598},
note = {CC BY},
}
RIS
TY - JOUR
TI - HIGH-THROUGHPUT SCREENING OF BLOOD–BRAIN BARRIER-PENETRANT COMPOUNDS IDENTIFIES THERAPEUTIC CANDIDATES FOR CENTRAL NERVOUS SYSTEM-TROPIC PEDIATRIC ACUTE LYMPHOBLASTIC LEUKEMIA
AU - Manuela Eduarda FRANÇA
AU - Jessica Oliveira DE SANTIS
AU - Mariane Minussi BAPTISTELLA
AU - Leandro Henrique DE PAULA ASSIS
AU - Veridiana Kiill SUAZO
AU - Rosane Gomes DE PAULA QUEIROZ
AU - José Andrés YUNES
AU - Graziella Ribeiro SOUSA
AU - Carlos Alberto SCRIDELI
PY - 2026
DA - 2026/10/23
JO - Hematology, Transfusion and Cell Therapy
DO - 10.1016/j.htct.2026.107598
UR - https://doi.org/10.1016/j.htct.2026.107598
AB - Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy, accounting for approximately 80% of childhood leukemia cases. Despite therapeutic advances, central nervous system (CNS) involvement remains a major clinical challenge, associated with chemoresistance and relapse, partly due to the limited ability of conventional chemotherapeutic agents to cross the blood–brain barrier (BBB). To identify drug repurposing candidates for pediatric ALL, 270 compounds from the CNS-Penetrant Compound Library were screened using the Ex vivo Leukemia Drug Advisor (ELDA) platform, which combines automated real-time imaging and computational analysis. Cells were continuously monitored for 48 hours, enabling the generation of chemosensitivity curves according to drug concentration and exposure time. All evaluated compounds, including library compounds and the positive controls methotrexate, vincristine, and cytarabine, were tested at a fixed concentration of 27 µM. Assay quality was validated by Z-factor analysis, with values ≥0.7 across all evaluated cell lines (Jurkat, Nalm6, RS4;11, SUP-B15, and K562), confirming the robustness and reproducibility of the platform. Hits were identified based on compounds inducing ≥50% reduction in cell viability, followed by integrated analysis to select candidates showing consistent activity across all five cell lines at both 24 and 48 hours of treatment. This strategy identified 16 candidate compounds. Among these, BMS-202, BAY 11-7082, and WP1066 were prioritized based on their translational potential, BBB-penetrating properties, and limited evidence regarding their therapeutic application in pediatric ALL. These compounds were selected for further pharmacological evaluation, including dose–response experiments to determine IC₅₀ values in leukemic cell lines, which are currently underway. To evaluate cytotoxicity in non-malignant cells, two bone marrow-derived cell lines, MSC-hTERT and HS-5, were included. The IC₅₀ values obtained to date after 48 hours of treatment were 37.0 μM and 14.2 μM for BMS-202, 13.5 μM and 7.2 μM for BAY 11-7082, and 15.9 μM and 10.0 μM for WP1066 in MSC-hTERT and HS-5 cells, respectively. Collectively, these findings demonstrate the applicability of a BBB-penetrant compound-based screening strategy for identifying drug repurposing candidates in pediatric ALL. The identification of BMS-202, BAY 11-7082, and WP1066 highlights the potential for new therapeutic applications of molecules that remain poorly explored in this context, expanding treatment opportunities for a disease with unmet clinical needs. These results reinforce the relevance of investigating BBB-penetrant compounds and support further pharmacological and functional characterization of these candidates, particularly for therapeutic strategies targeting patients with CNS involvement. Funding: National Council for Scientific and Technological Development (CNPq), Process No. 168989/2023-9; São Paulo Research Foundation (FAPESP), Process No. 2022/09037-3; Coordination for the Improvement of Higher Education Personnel (CAPES), Finance Code 001; Graduate Program of the Department of Genetics, Ribeirão Preto Medical School, University of São Paulo; Teaching, Research and Assistance Support Foundation of the Hospital das Clínicas, Ribeirão Preto Medical School, University of São Paulo (FAEPA).
SN - 2531-1379
SN - 2531-1387
ER -