• Non-small cell lung cancer
• antibody-drug conjugates
• chemotherapy
• immune checkpoint inhibitors
• microtubule-targeting agents
• phase III trial
• taxanes
• vinca alkaloid

• Humans
• Carcinoma, Non-Small-Cell Lung/drug therapy
• Carcinoma, Non-Small-Cell Lung/pathology
• Lung Neoplasms/drug therapy
• Tubulin Modulators/therapeutic use
• Antineoplastic Agents/therapeutic use
• Immune Checkpoint Inhibitors/therapeutic use
• Immunotherapy/methods
• Animals
• Antineoplastic Combined Chemotherapy Protocols/therapeutic use
• Antineoplastic Combined Chemotherapy Protocols/pharmacology

• TAM receptors
• actin remodeling
• endothelial cell biology
• protein S
• vascular endothelial cadherin
• vascular permeability

• LNCC86-2021 Ligue National Contre le Cancer

• antitumor growth
• focal adhesion kinase
• hepatocellular carcinoma
• patient-derived xenograft
• sorafenib
• vinorelbine

• Humans
• Animals
• Mice
• Sorafenib/pharmacology
• Sorafenib/therapeutic use
• Carcinoma, Hepatocellular/metabolism
• Vinorelbine/pharmacology
• Liver Neoplasms/metabolism
• Antineoplastic Combined Chemotherapy Protocols/pharmacology
• Apoptosis
• Cell Line, Tumor
• Cell Proliferation
• Xenograft Model Antitumor Assays
• Antineoplastic Agents/pharmacology
• Antineoplastic Agents/therapeutic use

• NMRC/MOHIAFCAT2/006/2016 National Medical Research Council
• CG21APR1005 National University Hospital
• CGAug16M005 National University Hospital
• NRF-CRP17-2017-05 National Research Foundation
• AM/TP049/2021 SingHealth DUKE-NUS Academic Medicine Research Grant

• cancer
• endothelial cells
• heart
• hypertension
• mitotic inhibitors
• tubulin

• Humans
• Animals
• Dogs
• Cardiotoxicity
• Polymerization
• Neoplasms/drug therapy
• Antineoplastic Agents/toxicity
• Heart Diseases/chemically induced

• Yukiko Kuwata

• actin cortex
• bleb
• filopodium
• focal contacts
• vimentin intermediate filaments

• Animals
• Rats
• Cell Movement
• Cell Surface Extensions/metabolism
• Focal Adhesions/metabolism
• Intermediate Filaments/metabolism
• Microtubules/metabolism
• Vimentin/genetics
• Vimentin/metabolism
• Vimentin/pharmacology

• CA-4 analogs
• anticancer
• aqueous solubility
• cyano
• structural development

• Antineoplastic Agents/pharmacology
• Cell Proliferation
• Drug Screening Assays, Antitumor
• Molecular Structure
• Solubility
• Stilbenes/chemistry
• Structure-Activity Relationship
• Tubulin/metabolism
• Tubulin Modulators/pharmacology
• Humans
• Cell Line, Tumor

Photodynamic therapy (PDT) is a method in which a photosensitizer is administered in vivo and irradiated with light to generate reactive oxygen species (ROS), thereby causing the selective death of cancer cells. Since PDT is a noninvasive cancer treatment method with few adverse effects, it has attracted considerable attention and is increasingly used. In PDT, there are two dominant processes based on the actin filament (A-filament) formation effect: the destruction of cells by necrosis and vascular shutdown. Despite the importance of its fine control, the mechanism of the reaction process from the generation of reactive oxygen by photoinduction inducing the formation of A-filament and its polymerization to form stress fibers (S-fibers) has not yet been clarified because, for example, it has been difficult to directly observe and quantify such processes in living cells by conventional methods. Here, we have combined atomic force microscopy (AFM) with other techniques to reveal the mechanism of the A-filament and S-fiber formation processes that underlie the cell death process due to PDT. First, it was confirmed that activation of the small G protein RhoA, which is a signal that induces an increase in A-filament production, begins immediately after PDT treatment. The production of A-filament did not increase with increasing light intensity when the amount of light was large. Namely, the activation of RhoA reached an equilibrium state in about 1 min: however, the production of A-filament and its polymerization continued. The observed process corresponds well with the change in the amount of phosphorylated myosin-light chains, which induce A-filament polymerization. The increase in the elastic modulus of cells following the formation of S-fiber was confirmed by AFM for the first time. The distribution of generated A-filament and S-fiber was consistent with the photosensitizer distribution. PDT increases A-filament production, and when the ROS concentration is high, blebbing occurs and cells die, but when it is low, cell death does not occur and S-fiber is formed. That is, it is expected that vascular shutdown can be controlled efficiently by adjusting the amount of photosensitizer and the light intensity.

We have combined atomic force microscopy with other techniques to reveal the mechanism of the actin filament and stress fibers formation processes that underlies the cell death process due to photodynamic therapy.

• metalloproteins
• drug discovery and development

• Animals
• Antineoplastic Agents/chemistry
• Antineoplastic Agents/pharmacology
• Biocatalysis
• Biological Phenomena
• Cell Line, Tumor
• Drug Delivery Systems
• Drug Design
• Drug Development
• Drug Discovery
• Female
• Humans
• Infant, Newborn
• Infant, Newborn, Diseases
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Molecular Docking Simulation
• Neoplasms/drug therapy
• Prodrugs/chemistry
• Xenograft Model Antitumor Assays

• Japan Agency for Medical Research and Development (AMED)
• Mizutani Foundation for Glycoscience (Mizutani Foundation)
• MEXT | Japan Society for the Promotion of Science (JSPS)

• Microtubules
• angiogenesis
• chalcone
• fluorinated
• zebrafish

Vascular-disrupting agents promise significant therapeutic efficacy against solid tumors by selectively damaging tumor-associated vasculature. Dynamic BLI and oxygen-enhanced multispectral optoacoustic tomography (OE-MSOT) were used to compare vascular shutdown following administration of KGP265. BLI signal and vascular oxygenation response (ΔsO₂) to a gas breathing challenge were both significantly reduced within 2 h indicating vascular disruption, which continued over 24 h. Twice-weekly doses of KGP265 caused a significant growth delay in MDA-MB-231 human breast tumor xenografts and 4T1 syngeneic breast tumors growing orthotopically in mice.

The selective disruption of tumor-associated vasculature represents an attractive therapeutic approach. We have undertaken the first in vivo evaluation of KGP265, a water-soluble prodrug of a benzosuberene-based tubulin-binding agent, and found promising vascular-disrupting activity in three distinct tumor types. Dose escalation in orthotopic MDA-MB-231-luc breast tumor xenografts in mice indicated that higher doses produced more effective vascular shutdown, as revealed by dynamic bioluminescence imaging (BLI). In syngeneic orthotopic 4T1-luc breast and RENCA-luc kidney tumors, dynamic BLI and oxygen enhanced multispectral optoacoustic tomography (OE-MSOT) were used to compare vascular shutdown following the administration of KGP265 (7.5 mg/kg). The BLI signal and vascular oxygenation response (ΔsO₂) to a gas breathing challenge were both significantly reduced within 2 h, indicating vascular disruption, which continued over 24 h. A correlative histology confirmed increased necrosis and hemorrhage. Twice-weekly doses of KGP265 caused significant growth delay in both MDA-MB-231 and 4T1 breast tumors, with no obvious systemic toxicity. A combination with carboplatin produced significantly greater tumor growth delay than carboplatin alone, though significant carboplatin-associated toxicity was observed (whole-body weight loss). KGP265 was found to be effective at low concentrations, generating long-term vascular shutdown and tumor growth delay, thus providing strong rationale for further development, particularly in combination therapies.

• benzosuberene
• bioluminescence imaging (BLI)
• breast tumors
• carboplatin
• combretastatin
• kidney tumors
• multispectral optoacoustic tomography (MSOT)
• photoacoustics
• vascular-disrupting agent (VDA)

• R01 CA140674 NCI NIH HHS
• S10 OD018094 NIH HHS
• R01 CA244579 NCI NIH HHS
• P30 CA142543 NCI NIH HHS
• 5R01 CA140674 NCI NIH HHS
• S10 RR024757 NCRR NIH HHS
• U24 CA126608 NCI NIH HHS
• 1R01 CA244579-01A1 NCI NIH HHS

• Eph
• EphA4
• motor neuron death
• motor neuron disease
• motor neurons
• neurodegenerative disease
• pathogenesis
• receptors

• Animals
• Cell Death
• Humans
• Motor Neuron Disease/metabolism
• Motor Neuron Disease/pathology
• Motor Neurons/metabolism
• Motor Neurons/pathology
• Receptor, EphA4/metabolism
• Signal Transduction

• TRG 2018 Bartlett FightMND

• bioluminescence
• breast cancer
• combretastatins
• imaging
• inhibitors of tubulin polymerization
• kidney cancer
• lung cancer
• magnetic resonance imaging
• photoacoustics
• vascular disrupting agents

• Antineoplastic Agents/therapeutic use
• Cell Death/drug effects
• Cell Proliferation/drug effects
• Humans
• Necrosis/drug therapy
• Necrosis/genetics
• Necrosis/pathology
• Neoplasms/drug therapy
• Neoplasms/genetics
• Neovascularization, Pathologic/drug therapy
• Neovascularization, Pathologic/genetics
• Neovascularization, Pathologic/pathology
• Protein Binding
• Tubulin/drug effects
• Tubulin/genetics
• Tubulin Modulators/chemistry
• Tubulin Modulators/therapeutic use

• RP140399 Cancer Prevention and Research Institute of Texas
• R01 CA140674 NCI NIH HHS
• 1R01CA244579-01A1 NCI NIH HHS
• S10 RR024757 NCRR NIH HHS
• S10 RR031859 NIH HHS
• R01 CA244579 NCI NIH HHS
• P30 CA142543 NCI NIH HHS
• RP100406 Cancer Prevention and Research Institute of Texas
• 1U24 CA126608 NCI NIH HHS
• S10 RR025648 NCRR NIH HHS
• S10 OD018094 NIH HHS
• S10 OD018094-01A1 NIH HHS
• 5R01 CA140674 NCI NIH HHS
• 1S10 RR024757 NIH HHS

• Cancer
• Critical cysteine residues
• Glutathione
• Oxidative stress
• Redox balance
• S-Glutathionylation

Adenosine triphosphate (ATP) is among the molecules involved in the immune response. It acts as danger signal that promotes inflammation by activating both P2X and P2Y purinergic receptors expressed in immune cells, including microglia, and tumor cells. One of the most important receptors implicated in ATP-induced inflammation is P2X7 receptor (P2X7R). The stimulation of P2X7R by high concentration of ATP results in cell proliferation, inflammasome activation and shedding of extracellular vesicles (EVs). EVs are membrane structures released by all cells, which contain a selection of donor cell components, including proteins, lipids, RNA and ATP itself, and are able to transfer these molecules to target cells. ATP stimulation not only promotes EV production from microglia but also influences EV composition and signaling to the environment. In the present review, we will discuss the current knowledge on the role of ATP in the biogenesis and dynamics of EVs, which exert important functions in physiology and pathophysiology.

• ATP
• ATP cargo
• P2X7 receptor
• extracellular vesicle biogenesis
• extracellular vesicle morphology
• extracellular vesicles
• immune cells
• tumor cells

• Microtubule inhibitor
• SCB01A
• Solid tumor

• Rho-GTP pathway
• photodynamic therapy
• talaporfin sodium
• vascular shutdown effect

• IMB5046
• endothelial cells
• focal adhesion kinase
• membrane blebbing
• microtubule-depolymerizing agents
• stress fibres

• Benzoates/pharmacology
• Cardiac Myosins/metabolism
• Cell Surface Extensions/drug effects
• Cell Surface Extensions/metabolism
• Cytoskeleton/drug effects
• Cytoskeleton/metabolism
• Endothelial Cells/drug effects
• Endothelial Cells/metabolism
• Enzyme Activation/drug effects
• Focal Adhesion Protein-Tyrosine Kinases/metabolism
• Focal Adhesions/drug effects
• Focal Adhesions/metabolism
• Heat-Shock Proteins/metabolism
• Humans
• Integrins/metabolism
• Microtubules/metabolism
• Models, Biological
• Molecular Chaperones/metabolism
• Morpholines/pharmacology
• Myosin Light Chains/metabolism
• Phosphorylation/drug effects
• Protein Kinase Inhibitors/pharmacology
• Quinolones/pharmacology
• Rho Guanine Nucleotide Exchange Factors/metabolism
• Signal Transduction/drug effects
• Stress Fibers/drug effects
• Stress Fibers/metabolism
• Sulfones/pharmacology
• rho GTP-Binding Proteins/metabolism
• rho-Associated Kinases/metabolism

• National Natural Science Foundation of China

Combretastatin A-4 phosphate (CA4P) is a microtubule-disrupting tumour-selective vascular disrupting agent (VDA). CA4P activates the actin-regulating RhoA-GTPase/ ROCK pathway, which is required for full vascular disruption. While hypoxia renders tumours resistant to many conventional therapies, little is known about its influence on VDA activity. Here, we found that active RhoA and ROCK effector phospho-myosin light chain (pMLC) were downregulated in endothelial cells by severe hypoxia. CA4P failed to activate RhoA/ROCK/pMLC but its activity was restored upon reoxygenation. Hypoxia also inhibited CA4P-mediated actinomyosin contractility, VE-cadherin junction disruption and permeability rise. Glucose withdrawal downregulated pMLC, and coupled with hypoxia, reduced pMLC faster and more profoundly than hypoxia alone. Concurrent inhibition of glycolysis (2-deoxy-D-glucose, 2DG) and mitochondrial respiration (rotenone) caused profound actin filament loss, blocked RhoA/ROCK signalling and rendered microtubules  CA4P-resistant. Withdrawal of the metabolism inhibitors restored the cytoskeleton and CA4P activity. The AMP-activated kinase AMPK was investigated as a potential mediator of pMLC downregulation. Pharmacological AMPK activators that generate AMP, unlike allosteric activators, downregulated pMLC but only when combined with 2DG and/or rotenone. Altogether, our results suggest that Rho/ROCK and actinomyosin contractility are regulated by AMP/ATP levels independently of AMPK, and point to hypoxia/energy depletion as potential modifiers of CA4P response.

• Antineoplastic Agents/pharmacology
• Breast Neoplasms/drug therapy
• Cell Line, Tumor
• Cell Proliferation/drug effects
• Colchicine/pharmacology
• Fatty Acids/pharmacology
• HEK293 Cells
• Humans
• Lung Neoplasms/drug therapy
• Molecular Structure
• Stilbenes/chemistry
• Stilbenes/pharmacology
• Tubulin Modulators/pharmacology

It is over 50 years since the discovery of microtubules, and they have become one of the most important drug targets for anti-cancer therapies. Microtubules are predominantly composed of the protein tubulin, which contains a number of different binding sites for small-molecule drugs. There is continued interest in drug development for compounds targeting the colchicine-binding site of tubulin, termed colchicine-binding site inhibitors (CBSIs). This review highlights CBSIs discovered through diverse sources: from natural compounds, rational design, serendipitously and via high-throughput screening. We provide an update on CBSIs reported in the past three years and discuss the clinical status of CBSIs. It is likely that efforts will continue to develop CBSIs for a diverse set of cancers, and this review provides a timely update on recent developments.

• CA-4
• anti-cancer
• anti-tubulin
• colchicine
• colchicine binding site
• combretastatin
• microtubule-targeting agents
• tubulin binding
• tubulin destabilizing

• Antiangiogenesis therapy
• Core@shell nanostructures
• Gold nanobipyramids
• Photothermal therapy
• Plasmon resonance

• chimeric antigen receptor
• combretastatin A-4 phosphate
• solid tumors
• vascular disrupting agent

• Alkaline hydrolysis
• Biodistribution
• Pharmacokinetics
• Poly (l-glutamic acid)-CA4 conjugate
• UHPLC-MS/MS

• Animals
• Antineoplastic Agents/chemistry
• Antineoplastic Agents/pharmacokinetics
• Chromatography, High Pressure Liquid/methods
• Glutamic Acid/chemistry
• Glutamic Acid/pharmacokinetics
• Hydrolysis
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Nanomedicine/methods
• Polyethylene Glycols/chemistry
• Polyethylene Glycols/pharmacokinetics
• Stilbenes/chemistry
• Stilbenes/pharmacokinetics
• Tandem Mass Spectrometry/methods
• Tissue Distribution

• Hematology
• Lung cancer
• Oncology
• Vasculitis
• hypoxia

New inhibitors of tubulin polymerization and/or histone deacetylase (HDAC) activity were synthesized by attaching alkyl tethered hydroxamic acid appendages of varying length to oxazole-bridged combretastatin A-4 analogous caps. While their antiproliferative and microtubule disrupting effect was most pronounced for derivatives with short spacers, HDAC inhibition was strongest for those with longer spacers. These findings were further supported by computational methods such as structure-based docking experiments exploring the target interactions of the derivatives with varying linkers. For instance, compounds featuring short four-atom spacers between cap and hydroxamic acid inhibited the growth of various cancer cell lines and human endothelial hybrid cells with IC₅₀ values in the low nanomolar range. In line with their ability to inhibit the microtubule assembly, four- and five-atom spacered hydroxamic acids caused an accumulation of 518A2 melanoma cells in G2/M phase, whereas a compound featuring a six-atom spacer and performing best in HDAC inhibition, induced a G1 arrest in these cells. All these beneficial anticancer activities together with their selectivity for cancer cells over non-malignant cells, point out the great potential of these novel pleiotropic HDAC and tubulin inhibitors as drug candidates for cancer therapy.

• anticancer agents
• combretastatin A-4
• histone deacetylase
• oxazole
• tubulin

• anticancer agents
• palladium catalysis
• sydnone
• tubulin binders
• vascular disrupting agents

• molecular mechanisms
• sulfonamides
• tubulin binding drugs
• tubulin binding sites

• active targeting nanoparticles
• combretastatin A4 (CA-4)
• crosslinked multilamellar liposomal vesicle
• doxorubicin (Dox)
• nanomedicine
• targeted drug combination therapy

• Angiogenesis inhibitors
• Cancer treatment
• Endothelial cells
• Plocabulin (PM060184)
• Tubulin inhibitors
• Vascular-disrupting agents

Vinca alkaloids, the well-known tubulin-binding agents, are widely used for the clinical treatment of malignant tumors. However, little attention has been paid to their vascular disrupting effects, and the underlying mechanisms remain largely unknown. This study aims to investigate the vascular disrupting effect and the underlying mechanisms of vinca alkaloids.

Methods: The capillary disruption assay and aortic ring assay were performed to evaluate the in vitro vascular disrupting effect of desacetylvinblastine monohydrazide (DAVLBH), a derivate of vinblastine, and the in vivo vascular disrupting effect was assessed on HepG2 xenograft model using magnetic resonance imaging, hematoxylin and eosin staining and immunohistochemistry. Tubulin polymerization, endothelial cell monolayer permeability, western blotting and immunofluorescence assays were performed to explore the underlying mechanisms of DAVLBH-mediated tumor vascular disruption.

Results: DAVLBH has potent vascular disrupting activity both in vitro and in vivo. DAVLBH disrupts tumor vessels in a different manner than classical tubulin-targeting VDAs; it inhibits microtubule polymerization, promotes the internalization of vascular endothelial cadherin (VE-cadherin) and inhibits the recycling of internalized VE-cadherin to the cell membrane, thus increasing endothelial cell permeability and ultimately resulting in vascular disruption. DAVLBH-mediated promotion of VE-cadherin internalization and inhibition of internalized VE-cadherin recycling back to the cell membrane are partly dependent on inhibition of microtubule polymerization, and Src activation is involved in DAVLBH-induced VE-cadherin internalization.

Conclusions: This study sheds light on the tumor vascular disrupting effect and underlying mechanisms of vinca alkaloids and provides new insight into the molecular mechanism of tubulin-targeting VDAs.

• VE-cadherin internalization.
• Vascular disrupting agents
• desacetylvinblastine monohydrazide
• tubulin polymerization
• vinca alkaloids

We investigated the microtubule-destabilizing, vascular-targeting, anti-tumor and anti-metastatic activities of a new series of chalcones, whose prototype compound is (E)-3-(3’’-amino-4’’-methoxyphenyl)-1-(5’-methoxy-3’,4’-methylendioxyphenyl)-2-methylprop-2-en-1-one (TUB091). X-ray crystallography showed that these chalcones bind to the colchicine site of tubulin and therefore prevent the curved-to-straight structural transition of tubulin, which is required for microtubule formation. Accordingly, TUB091 inhibited cancer and endothelial cell growth, induced G2/M phase arrest and apoptosis at 1-10 nM. In addition, TUB091 displayed vascular disrupting effects in vitro and in the chicken chorioallantoic membrane (CAM) assay at low nanomolar concentrations. A water-soluble L-Lys-L-Pro derivative of TUB091 (i.e. TUB099) showed potent antitumor activity in melanoma and breast cancer xenograft models by causing rapid intratumoral vascular shutdown and massive tumor necrosis. TUB099 also displayed anti-metastatic activity similar to that of combretastatin A4-phosphate. Our data indicate that this novel class of chalcones represents interesting lead molecules for the design of vascular disrupting agents (VDAs). Moreover, we provide evidence that our prodrug approach may be valuable for the development of anti-cancer drugs.

• cancer
• drug research
• tubulin
• vascular-disrupting

• VE-cadherin
• adherens junctions
• combretastatin
• sphingosine-1-phosphate
• tumour microcirculation

• bioavailability
• cancer therapy
• combretastatin A4 phosphate
• nanoparticles
• oral absorption

• Combretastatin A4
• Delivery
• Glutamic acid
• Nanomedicine
• Tumor therapy
• Vascular disrupting agents

• Animals
• Antineoplastic Agents, Phytogenic/administration & dosage
• Antineoplastic Agents, Phytogenic/pharmacokinetics
• Biocompatible Materials/chemistry
• Cell Line, Tumor
• Drug Delivery Systems
• Male
• Materials Testing
• Mice
• Mice, Inbred BALB C
• Mice, Nude
• Nanoconjugates/administration & dosage
• Nanoconjugates/chemistry
• Nanotechnology
• Neoplasms, Experimental/drug therapy
• Neoplasms, Experimental/metabolism
• Neoplasms, Experimental/pathology
• Polyglutamic Acid/administration & dosage
• Rats, Wistar
• Stilbenes/administration & dosage
• Stilbenes/pharmacokinetics

The cis-stilbene, combretastatin A4 (CA4), is a potent microtubule targeting and vascular damaging agent. Despite promising results at the pre-clinical level and extensive clinical evaluation, CA4 has yet to be approved for therapeutic use. One impediment to the development of CA4 is an inherent conformational instability about the ethylene linker, which joins two aromatic rings. We have previously published preliminary data regarding structurally simplified biphenyl derivatives of CA4, lacking an ethylene linker, which retain anti-proliferative and pro-apoptotic activity, albeit at higher doses. Our current study provides a more comprehensive evaluation regarding the anti-proliferative and pro-apoptotic properties of biphenyl CA4 derivatives in both 2D and 3D cancerous and non-cancerous cell models. Computational analysis has revealed that cytotoxicity of CA4 and biphenyl analogues correlates with predicted tubulin affinity. Additional mechanistic evaluation of the biphenyl derivatives found that their anti-cancer activity is dependent on prolonged mitotic arrest, in a similar manner to CA4. Lastly, we have shown that cancer cells deficient in the extrinsic pathway of apoptosis experience delayed cell death following treatment with CA4 or analogues. Biphenyl derivatives of CA4 represent structurally simplified analogues of CA4, which retain a similar mechanism of action. The biphenyl analogues warrant in vivo examination to evaluate their potential as vascular damaging agents.

• Antineoplastic Agents/chemistry
• Antineoplastic Agents/metabolism
• Antineoplastic Agents/pharmacology
• Apoptosis/drug effects
• Biphenyl Compounds/chemistry
• Cell Cycle Checkpoints/drug effects
• Cell Proliferation/drug effects
• DNA Damage
• Humans
• Jurkat Cells
• Membrane Potential, Mitochondrial/drug effects
• Mitosis/drug effects
• Models, Molecular
• Protein Conformation
• Stilbenes/chemistry
• Stilbenes/metabolism
• Stilbenes/pharmacology
• Tubulin/chemistry
• Tubulin/metabolism

• Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada (CA)
• NSERC

• Amoeboid movement
• Anticancer therapy
• Cell adhesion
• Collective migration
• Epithelial-to-mesenchymal transition
• Mesenchymal migration
• Metastasis
• Plasticity
• Proteolysis
• Tumor cell invasion