Cancer stem cells (CSCs) are increasingly recognized as key players in tumor development and recurrence. These cells, characterized by stem-like properties, exhibit persistent activation of critical signaling pathways such as Notch, Hedgehog (HH), and Wnt, which are essential for development and tissue homeostasis. CSCs are resistant to conventional therapies and have slow growth rates, making them a challenging target for cancer treatment. Recent advances in targeting these pathways aim to control CSC replication, survival, and differentiation. This article provides an update on the clinical development of agents targeting the Notch, HH, and Wnt pathways, discussing strategies to overcome resistance and improve treatment outcomes. The Notch pathway, a conserved cell-fate-determination mechanism, is involved in various aspects of cancer biology, including CSCs and tumor immunity. Notch signaling is mediated by transmembrane ligands and receptors, leading to the release of an intracellular fragment that regulates gene expression. γ-secretase inhibitors (GSIs) are a major class of Notch pathway inhibitors, showing antineoplastic activity in preclinical models and clinical trials. However, GSIs are associated with significant side effects, such as secretory diarrhea and cutaneous rash. Strategies to mitigate these effects include intermittent dosing and combination with glucocorticoids or anti-oestrogen agents. Anti-DLL4 antibodies and other Notch inhibitors are also under investigation. The HH pathway is involved in tissue patterning and repair, and its dysregulation contributes to various cancers, including basal-cell carcinoma (BCC) and medulloblastoma. Vismodegib, a direct SMO inhibitor, is the most clinically advanced HH pathway inhibitor, approved for BCC treatment. However, its efficacy in other cancers is limited, and resistance mechanisms, such as mutations in SMO or GLI transcription factors, are being explored. Combination therapies with other pathways, such as PI3K-AKT, are being investigated to enhance efficacy. The Wnt pathway, particularly the canonical Wnt pathway, is crucial for tumor development and progression. Inhibitors of this pathway, such as porcupine inhibitors, are being developed to target CSCs. The development of Wnt pathway inhibitors is also being explored in combination with other therapies to overcome resistance and improve outcomes. In summary, targeting the Notch, HH, and Wnt pathways in CSCs is a promising approach for cancer treatment. However, challenges such as resistance, side effects, and the need for biomarkers to guide treatment remain. Ongoing research aims to develop more effective and safer agents, leveraging combination therapies and novel mechanisms of action to improve clinical outcomes.Cancer stem cells (CSCs) are increasingly recognized as key players in tumor development and recurrence. These cells, characterized by stem-like properties, exhibit persistent activation of critical signaling pathways such as Notch, Hedgehog (HH), and Wnt, which are essential for development and tissue homeostasis. CSCs are resistant to conventional therapies and have slow growth rates, making them a challenging target for cancer treatment. Recent advances in targeting these pathways aim to control CSC replication, survival, and differentiation. This article provides an update on the clinical development of agents targeting the Notch, HH, and Wnt pathways, discussing strategies to overcome resistance and improve treatment outcomes. The Notch pathway, a conserved cell-fate-determination mechanism, is involved in various aspects of cancer biology, including CSCs and tumor immunity. Notch signaling is mediated by transmembrane ligands and receptors, leading to the release of an intracellular fragment that regulates gene expression. γ-secretase inhibitors (GSIs) are a major class of Notch pathway inhibitors, showing antineoplastic activity in preclinical models and clinical trials. However, GSIs are associated with significant side effects, such as secretory diarrhea and cutaneous rash. Strategies to mitigate these effects include intermittent dosing and combination with glucocorticoids or anti-oestrogen agents. Anti-DLL4 antibodies and other Notch inhibitors are also under investigation. The HH pathway is involved in tissue patterning and repair, and its dysregulation contributes to various cancers, including basal-cell carcinoma (BCC) and medulloblastoma. Vismodegib, a direct SMO inhibitor, is the most clinically advanced HH pathway inhibitor, approved for BCC treatment. However, its efficacy in other cancers is limited, and resistance mechanisms, such as mutations in SMO or GLI transcription factors, are being explored. Combination therapies with other pathways, such as PI3K-AKT, are being investigated to enhance efficacy. The Wnt pathway, particularly the canonical Wnt pathway, is crucial for tumor development and progression. Inhibitors of this pathway, such as porcupine inhibitors, are being developed to target CSCs. The development of Wnt pathway inhibitors is also being explored in combination with other therapies to overcome resistance and improve outcomes. In summary, targeting the Notch, HH, and Wnt pathways in CSCs is a promising approach for cancer treatment. However, challenges such as resistance, side effects, and the need for biomarkers to guide treatment remain. Ongoing research aims to develop more effective and safer agents, leveraging combination therapies and novel mechanisms of action to improve clinical outcomes.