Background: The development of novel tubulin-targeting agents remains a promising strategy in anticancer drug design. In the present study, novel heterocyclic tubulin-targeting compounds were identified using a complex in silico/in vitro workflow. Methods: An in-house library of approximately 8000 structurally diverse heterocyclic compounds was screened for antiproliferative activity, yielding 260 experimentally validated hits. These compounds were docked against the colchicine, vinca, and taxane binding sites of tubulin. Selected compounds were also tested for their effects on cellular β-tubulin protein levels in Jurkat cells. Results: Docking analysis indicated that 42 compounds matched or exceeded colchicine's binding energy, 3 compounds were comparable to vinblastine, and 18 compounds surpassed paclitaxel, suggesting the colchicine and taxane sites as preferred targets. Five top-scoring compounds (Les-6370, Les-4272, Les-4309, Les-4403, and Les-1419) were further analyzed via molecular dynamics simulations, confirming stable binding-particularly at the colchicine site-and favorable hydrogen-bonding and lipophilic interactions. At 10 µM, Les-1419 and Les-4309 reduced total cellular β-tubulin protein levels in Jurkat cells by 57.73% and 54.31%, respectively, relative to the effect observed for the reference compound Combretastatin A-4. Conclusions: This study resulted in the identification of novel starting scaffolds with favorable tubulin interaction activity and predicted drug-like properties, representing structurally novel parent structures and serving as promising starting points for further structural optimization and analogue development.

Purpose: Chemotherapy can be a viable option for patients with relapsed-refractory melanoma (RRM) who do not have access to advanced treatments or clinical trials. We report our experience with a modified Vinblastine-Cisplatin-Temozolomide (mVCT) regimen in patients with RRM and barriers to their enrollment in clinical trials. Methods: We conducted a retrospective review of patients with RRM who received mVCT. The primary endpoint was overall response rate (ORR) following the last cycle. Secondary endpoints included the rate of complete response (CR), partial response (PR), stable disease (SD), clinical benefit rate (CBR), progression-free survival (PFS), intracranial (IC) PFS, (IC) CBR, (IC) RR, and toxicity. Results: Ten patients received mVCT for a median of 4.5 cycles (IQR: 3-6). Median follow-up was 20 months (IQR: 10.3-30.3). The ORR was 50% (CR-1, PR-4) and CBR was 70%. The median PFS was 7 months (IQR: 1.5-9.25). Among eight patients with brain metastasis, six underwent gamma-knife radiosurgery (GKRS); three achieved PR, and three had SD. Two patients without GKRS achieved CR, leading to an ICRR of 62.5% and an ICCBR of 100%. The median ICPFS was 7.5 months (IQR: 6-9.75). Median OS was 22 months (95% CI: 15.6-32.4). Four patients experienced grade ≥ 3 toxicities (fatigue in two, myelosuppression in two); two required dose reduction and one discontinued treatment. Two patients with PD later enrolled in phase 1 clinical trials. Brain metastasis (80%), immunotherapy-related toxicity (60%), acral/mucosal melanoma (50%), and social barriers (40%) were the major reasons for trial ineligibility. Conclusions: mVCT is an effective and well-tolerated regimen for patients with RRM and can serve as a bridge while they await access to advanced treatments or a clinical trial.

Cancer remains a leading cause of mortality worldwide. The success of plant-derived chemotherapeutic agents, such as paclitaxel and vinblastine, underscores the significant potential of plant-derived natural products (PDNPs) in oncology. Despite this success, the poor solubility, metabolic instability, and nonspecific distribution of many PDNPs pose a fundamental barrier to their full clinical potential. Plant-derived extracellular vesicles (PDEVs) have garnered considerable attention as biocompatible nanocarriers, and numerous reviews have examined their biological activities and delivery applications. However, few have systematically addressed their synergistic combination with PDNPs in cancer therapy. This review distinguishes itself by centering on the synergistic interplay between PDNPs and PDEVs as an integrated therapeutic strategy. Specifically, we examine: (1) how PDEVs enhance the bioavailability and anticancer efficacy of PDNPs by improving their solubility, stability, and targeted delivery, and (2) how the intrinsic antitumor activity of PDEVs complements that of PDNPs to achieve enhanced therapeutic efficacy and reduced systemic toxicity compared with either agent alone. By integrating current evidence on the combined use of PDNPs and PDEVs, this review aims to enhance the field's understanding of the progress made by PDNPs and PDEVs in cancer therapy, clarify their mechanisms of action, and identify the critical challenges facing the current research stage.

Langerhans cell histiocytosis (LCH) is an inflammatory myeloid neoplasm driven by somatic variants in MAPK-encoding genes, most commonly BRAFV600E. The prognostic value of BRAFV600E in the peripheral blood of children with LCH has never been investigated in an international setting characterized by different approaches. To this aim, five centers from the European Consortium for Histiocytosis (ECHO) analyzed the peripheral blood of 226 children enrolled in, or managed according to, the LCH-IV protocol. We quantified BRAFV600E burden in different DNA sources at diagnosis and after 6 weeks of vinblastine/prednisone and evaluated its association with 2-year event-free survival (EFS). At diagnosis, BRAFV600E positivity could be equally determined through circulating cell-free (ccf) or peripheral blood mononuclear cell (PBMC)-DNA in patients with multi-system (MS) LCH. In single-system (SS) LCH, analyzing both substrates improved diagnostic sensitivity. For each clinical subgroup at diagnosis, we identified BRAFV600E levels associated with inferior 2-year EFS: ≥LOQ (0.1% for ccf-DNA, 0.01% for PBMC-DNA) in SS and MS risk-organ negative (RO-) groups; ≥1% (both ccf- and PBMC-DNA) in MS risk-organ positive (RO+) group. Notably, BRAFV600E burden was independently associated with inferior EFS in multivariable analysis (HR 2.05, 95%CI: 1.11-3.78 for BRAFV600E ≥LOQ, p=0.022; HR 4.53, 95%CI: 2.34-8.79 for BRAFV600E≥1%, p<0.001). After 6 weeks of vinblastine/prednisone, decreasing BRAFV600E burden (<0.1% in ccf-DNA or <0.01% in PBMC-DNA) was significantly associated with higher 2-year EFS (p<0.001 and p=0.009). These findings provide a framework for refining risk-stratification in pediatric LCH through the incorporation of peripheral blood BRAFV600E testing into international prospective clinical trials.

Single-system single-site Langerhans cell histiocytosis (SS-s LCH) can involve the spine and cause local neurological and structural complications. A 7-year-old boy with SS-s LCH of the fifth cervical vertebra presented with neck pain and torticollis. Owing to osteolytic destruction and concern for cord compression, the patient received 6 weeks of vinblastine and prednisolone. The lesion regressed, and treatment was discontinued after the local risk was reduced. Although an iliac recurrence developed later, the cervical lesion remained under control. This case suggests that response-adapted early discontinuation of chemotherapy may be considered in selected patients with vertebral SS-s LCH.

Frontline therapy for advanced-stage classical Hodgkin lymphoma (cHL) has entered a new era. Two phase III trials have established highly effective but conceptually different strategies: nivolumab plus doxorubicin, vinblastine, and dacarbazine (N-AVD) in SWOG S1826 and PET-adapted BrECADD in GHSG HD21. Because these regimens were tested against different comparators and in different populations, their outcomes should not be interpreted as a head-to-head comparison. This review focuses on translating both trials into individualized treatment selection. N-AVD combines excellent disease control with relatively low hematologic toxicity, minimal use of radiotherapy, operational simplicity, and direct evidence across a broad age range, including patients older than 60 years. BrECADD provides mature disease-control data, PET2-guided treatment duration, potentially only 4 cycles in patients with complete metabolic response, lower anthracycline exposure in PET2-negative patients, and substantially improved gonadal recovery compared with eBEACOPP. The relevant trade-offs differ: hematologic and infectious toxicity, immune-related adverse events, fertility, long-term toxicity, radiotherapy exposure, PET availability, and health-system logistics may each influence regimen selection. Rather than identifying a universal winner, contemporary practice should define which patient is best suited to each strategy and incorporate shared decision-making into frontline treatment.

Methyl jasmonate (MeJA) is a commonly used elicitor for enhancing secondary metabolite production, including the anticancer terpenoid indole alkaloids (TIAs) vinblastine and vincristine. To investigate the physiological and molecular responses associated with MeJA-induced TIA accumulation, we examined the effect of different MeJA concentrations on vinblastine and vincristine production, gene expression and antioxidant defence in Catharanthus roseus calli. MeJA significantly enhanced relative growth and increased vinblastine and vincristine accumulation, with the highest increases observed at 50 μM MeJA, reaching 159.2% and 183.4% of control, respectively, and upregulated CrMPK3, STR, and DAT expression. The lower concentrations of MeJA showed higher H2O2 accumulation, DPPH free radical-scavenging and flavonoid content. Whereas increased lipid peroxidation and PAL enzyme activity were associated with the higher concentrations (100 and 200 μM). The upregulated expression of CrMPK3, together with the biosynthetic genes STR and DAT, supports the involvement of MAPK in signalling responses that mediate MeJA-elicited TIAs. Pearson correlation and principal component analysis showed that H2O2 accumulation was more closely associated with vinblastine and vincristine accumulation than with the expression of the investigated genes at the experimental time points. These findings suggest that H2O2 accumulation may be associated with downstream metabolic/redox processes accompanying alkaloid accumulation, although an earlier transient role of H2O2 in MeJA signalling cannot be excluded. The enhanced flavonoid accumulation and DPPH scavenging activity further suggest a role for antioxidant responses in maintaining redox homeostasis during MeJA-induced alkaloid accumulation.

Catharanthus roseus is the sole natural source of the anticancer bisindole alkaloids vinblastine and vincristine, but leaf yields are intrinsically low. This study evaluated whether targeted in vitro and in vivo elicitation could enhance alkaloid accumulation and whether these changes were associated with selected physiological stress markers and with responses of terpenoid indole alkaloid-related transcripts. Microplants and calli were cultured on Murashige-Skoog medium; pot-grown plants received sodium chloride (NaCl), salicylic acid (10[-5] M), methyl jasmonate (MeJA; 500 µM), ultraviolet-B (UV-B), or MeJA plus UV-B under controlled pot-culture conditions. Vinblastine and vincristine were quantified by high-performance liquid chromatography; hydrogen peroxide, electrolyte leakage, proline, ascorbate, and chlorophyll were assayed; and transcript levels of strictosidine synthase (CrSTR), tryptophan decarboxylase (CrTDC), and deacetylvindoline O-acetyltransferase (CrDAT) were measured by quantitative reverse-transcription PCR. Elicitation significantly altered alkaloid accumulation (p < 0.001): callus formation/NaCl treatment 8 (CFNT8) reached 11.44 µg g[-1] dry weight (DW) vinblastine and 23.35 µg g[-1] DW vincristine. In the selected follow-up subset, high-alkaloid treatments were associated with higher hydrogen peroxide (2.41 µmol g[-1]), electrolyte leakage (0.44 µS cm[-1] g[-1]), proline (96 µg g[-1]), and ascorbate (26.14 mg g[-1]), lower chlorophyll, and increased CrSTR (14.3-fold), CrTDC (5.1-fold), and CrDAT (24.1-fold) transcript abundance. Exploratory principal component analysis explained 88.85% of the variance and indicated clustering of stress/antioxidant, transcript, and alkaloid traits. Because this analysis used a limited targeted subset, these patterns are interpreted as hypothesis-generating. The findings identify CFNT8 and selected UV-B/MeJA-responsive treatments as useful conditions for future validation of elicitation timing in C. roseus.

Rosai-Dorfman-Destombes disease (RDD) is a rare non-Langerhans cell histiocytosis with a broad clinical spectrum, ranging from localized lymphadenopathy to severe multisystem involvement. Due to its rarity, clinical data on pediatric RDD remain limited, particularly in North Africa. We report two contrasting pediatric cases of RDD managed at a tertiary center in Morocco to illustrate this clinical heterogeneity. The first was that of a five-year-old girl who presented with chronic, isolated, painless cervical lymphadenopathy. Laboratory findings were normal. Biopsy confirmed RDD with classic emperipolesis and an S100+, CD163+, CD68+, CD1a- immunophenotype. Targeted molecular analysis showed no mutations in KRAS, NRAS, or BRAF exon 15. Active surveillance led to progressive spontaneous regression. The second case was of a 20-month-old boy initially misdiagnosed with osteomyelitis due to painful bone lesions. He rapidly developed severe multisystem disease with axillary lymphadenopathy, respiratory distress secondary to pulmonary nodules, hepatomegaly, renal lesions, and widespread osteolytic destruction. Laboratory testing revealed a marked inflammatory syndrome, anemia, and polyclonal hypergammaglobulinemia. Subsequent lymph node biopsy established the diagnosis of RDD. Treatment with vinblastine and prednisone achieved long-term complete remission, with persistent bone deformity as the sole sequela. Pediatric RDD poses a significant diagnostic challenge due to its marked clinical variability. Histopathological confirmation showing emperipolesis and characteristic immunophenotyping is essential. Isolated nodal forms carry a favorable prognosis and can be managed conservatively, whereas severe multisystem disease requires prompt systemic therapy.

Madagascar periwinkle [Catharanthus roseus (L.) G. Don] produces vindoline and catharanthine, key precursors used in the semi-synthesis of the high-value anticancer alkaloids vinblastine and vincristine. Closed plant production systems (CPPS) allow precise application of environmental stimuli, including light spectrum and plasma-activated water (PAW). In a previous study, we demonstrated the potential of combining artificial light (white and red) and PAW to enhance the production of these alkaloids in periwinkle grown under controlled CPPS conditions. The present study expands this approach by adopting a more scalable PAW application method and a broader range of LED light spectra to evaluate their effects on vindoline and catharanthine accumulation. After a 37-day pre-treatment under white LEDs in environmentally controlled rooms, plants were subjected to fourteen treatments: phosphor-converted warm white (WW-P, control), red (R), blue (B), red-blue (RB), red-blue-far red (RBFR), phosphor-converted cool white (CW-P), and color-mixed white (W-CM) LED light, each with or without PAW. Plant growth and alkaloid-related responses were assessed at three sampling times: before treatment application (T0), 47 (T1), and 55 (T2) days after pre-treatment. RB produced the highest total vindoline and catharanthine yields [sum of leaf and root yields, with each organ yield calculated as organ dry mass (DM, g plant-1) × organ alkaloid concentration (µg g-1 DM)], reaching 228.0 ± 16.9 and 338.0 ± 27.0 µg plant-1, respectively, which were significantly higher than those obtained under all other light spectra (1.4-fold for vindoline and 1.3-fold for catharanthine relative to WW-P), except for W-CM, for which no significant differences were observed. PAW significantly increased total vindoline and catharanthine yields, reaching 211.0 ± 8.0 and 278.0 ± 11.0 µg plant-1, respectively, corresponding to 1.9- and 1.3-fold increases compared with the untreated condition. Sampling time had a strong effect on total vindoline and catharanthine yields, increasing from 6.0 ± 0.7 and 13.9 ± 1.7 µg plant-1, respectively, at T0, to 258.7 ± 9.8 and 414.0 ± 17.0 µg plant-1, respectively, at T2, corresponding to 43.1- and 29.8-fold increases. In conclusion, the combined use of RB light and PAW, together with harvesting at T2, represented the most effective strategy to achieve high vindoline and catharanthine production, as it enhanced the total yields of both alkaloids.

Microtubules are essential for cell division and intracellular signaling, making them well-established targets for anticancer therapy. However, the clinical utility of conventional microtubule-targeting agents is often limited by toxicity and drug resistance. SB225002, originally identified as an interleukin-8 receptor antagonist, has been reported to disrupt microtubule dynamics and exhibit anticancer activity with relatively low toxicity in preclinical studies. Yet, the structural basis of its interaction with tubulin remains poorly understood. Here, we determined the crystal structure of the SB225002-tubulin complex at 2.60 Å resolution and performed molecular dynamics simulations to characterize ligand binding, as well as drug-induced changes in tubulin structure and inter-subunit interactions. The crystal structure revealed three SB225002 molecules occupying the colchicine- and vinblastine-binding regions of tubulin. Simulations showed persistent ligand association over the simulated timescale and relatively limited overall changes in longitudinal inter-subunit interactions, whereas structural flexibility and dynamic coupling were preferentially altered at the lateral-interface-associated surface of β-tubulin. Additional simulations incorporating both lateral and longitudinal tubulin interfaces further revealed a greater reduction in lateral than in longitudinal inter-subunit contacts upon drug binding. Together, these results support a mechanism in which SB225002 preferentially perturbs lateral tubulin interactions and provide a structural framework for understanding its microtubule-modulating activity.