Because of synergism between tubulin and HDAC inhibitors, we used the pharmacophore fusion strategy to generate potential tubulin-HDAC dual inhibitors. Drug design was based on the introduction of a N-hydroxyacrylamide or a N-hydroxypropiolamide at the 5-position of the 2-aroylbenzo[b]furan skeleton, to produce compounds 6a-i and 11a-h, respectively. Among the synthesized compounds, derivatives 6a, 6c, 6e, 6g, 11a, and 11c showed excellent antiproliferative activity, with IC50 values at single- or double-digit nanomolar levels, against the A549, HT-29, and MCF-7 cells resistant towards the control compound combretastatin A-4 (CA-4). Compounds 11a and 6g were also 10-fold more active than CA-4 against the Hela cell line. When comparing the inhibition of tubulin polymerization versus the HDAC6 inhibitory activity, we found that 6a-g, 6i, 11a, 11c, and 11e, although very potent as inhibitors of tubulin assembly, did not have significant inhibitory activity against HDAC6.

• MeSH
• Benzofurans * pharmacology   chemistry   chemical synthesis   MeSH
• HT29 Cells MeSH
• HeLa Cells MeSH
• Histone Deacetylase 6 antagonists & inhibitors   metabolism   MeSH
• Histone Deacetylase Inhibitors pharmacology   chemical synthesis   chemistry   MeSH
• Hydroxamic Acids * pharmacology   chemistry   chemical synthesis   MeSH
• Humans MeSH
• MCF-7 Cells MeSH
• Tubulin Modulators * pharmacology   chemical synthesis   chemistry   MeSH
• Cell Line, Tumor MeSH
• Cell Proliferation * drug effects   MeSH
• Antineoplastic Agents * pharmacology   chemical synthesis   chemistry   MeSH
• Drug Screening Assays, Antitumor MeSH
• Tubulin * metabolism   MeSH
• Structure-Activity Relationship MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Microtubule dynamic is exceptionally sensitive to modulation by small-molecule ligands. Our previous work presented the preparation of microtubule-targeting estradiol dimer (ED) with anticancer activity. In the present study, we explore the effect of selected linkers on the biological activity of the dimer. The linkers were designed as five-atom chains with carbon, nitrogen or oxygen in their centre. In addition, the central nitrogen was modified by a benzyl group with hydroxy or methoxy substituents and one derivative possessed an extended linker length. Thirteen new dimers were subjected to cytotoxicity assay and cell cycle profiling. Dimers containing linker with benzyl moiety substituted with one or more methoxy groups and longer branched ones were found inactive, whereas other structures had comparable efficacy as the original ED (e.g. D1 with IC50 = 1.53 μM). Cell cycle analysis and immunofluorescence proved the interference of dimers with microtubule assembly and mitosis. The proposed in silico model and calculated binding free energy by the MM-PBSA method were closely correlated with in vitro tubulin assembly assay.

• MeSH
• Apoptosis MeSH
• Ethinyl Estradiol * chemistry   pharmacology   MeSH
• G2 Phase Cell Cycle Checkpoints drug effects   MeSH
• Microtubules MeSH
• Tubulin Modulators * chemistry   pharmacology   MeSH
• Cell Line, Tumor MeSH
• Antineoplastic Agents * chemistry   pharmacology   MeSH
• Triazoles * chemistry   pharmacology   MeSH
• Tubulin * metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Studying the anticancer activity of 5-arylidene-2-(4-hydroxyphenyl)aminothiazol-4(5H)-ones towards cell lines of different cancer types allowed the identification of hit-compounds inhibiting the growth of daunorubicin- (CEM-DNR, IC50 = 0.32-1.28 μM) and paclitaxel-resistant (K562-TAX, IC50 = 0.21-1.23 μM) cell lines, with favorable therapeutic indexes. The studied compounds induced apoptosis and cellular proliferation in treated CCRF-CEM cells. The hit compounds were shown to induce mitotic arrest by interacting with tubulin, inhibiting its polymerization by binding to the colchicine binding site.

• MeSH
• Apoptosis MeSH
• Tubulin Modulators * pharmacology   chemistry   MeSH
• Cell Line, Tumor MeSH
• Cell Proliferation MeSH
• Antineoplastic Agents * pharmacology   chemistry   MeSH
• Drug Screening Assays, Antitumor MeSH
• Tubulin metabolism   MeSH
• Binding Sites MeSH
• Structure-Activity Relationship MeSH
• Publication type
• Journal Article MeSH

Prostate cancer is primarily fatal after it becomes metastatic and castration-resistant despite novel combined hormonal and chemotherapeutic regimens. Hence, new therapeutic concepts and drug delivery strategies are urgently needed for the eradication of this devastating disease. Here we report the highly specific, in situ click chemistry driven pretargeted delivery of cytotoxic drug carriers to PSMA(+) prostate cancer cells. Anti-PSMA 5D3 mAb and its F(ab')2 fragments were functionalized with trans-cyclooctene (TCO), labeled with a fluorophore, and used as pretargeting components. Human serum albumin (ALB) was loaded with the DM1 antitubulin agent, functionalized with PEGylated tetrazine (PEG4-Tz), labeled with a fluorophore, and used as the drug delivery component. The internalization kinetics of components and the therapeutic efficacy of the pretargeted click therapy were studied in PSMA(+) PC3-PIP and PSMA(-) PC3-Flu control cells. The F(ab')2 fragments were internalized faster than 5D3 mAb in PSMA(+) PC3-PIP cells. In the two-component pretargeted imaging study, both components were colocalized in a perinuclear location of the cytoplasm of PC3-PIP cells. Better colocalization was achieved when 5D3 mAb was used as the pretargeting component. Consecutively, the in vitro cell viability study shows a significantly higher therapeutic effect of click therapy in PC3-PIP cells when 5D3 mAb was used for pretargeting, compared to its F(ab')2 derivative. 5D3 mAb has a longer lifetime on the cell surface, when compared to its F(ab')2 analogue, enabling efficient cross-linking with the drug delivery component and increased efficacy. Pretargeting and drug delivery components were cross-linked via multiple bioorthogonal click chemistry reactions on the surface of PSMA(+) PC cells forming nanoclusters, which undergo fast cellular internalization and intracellular transport to perinuclear locations.

• MeSH
• Albumins MeSH
• Antigens, Surface immunology   MeSH
• Click Chemistry methods   MeSH
• Cyclooctanes chemistry   MeSH
• Fluorobenzenes chemistry   MeSH
• Antineoplastic Agents, Phytogenic therapeutic use   MeSH
• Glutamate Carboxypeptidase II immunology   metabolism   MeSH
• Immunoglobulin Fab Fragments chemistry   metabolism   therapeutic use   MeSH
• Drug Delivery Systems methods   MeSH
• Humans MeSH
• Maytansine therapeutic use   MeSH
• Tubulin Modulators therapeutic use   MeSH
• Antibodies, Monoclonal chemistry   metabolism   therapeutic use   MeSH
• Cell Line, Tumor MeSH
• Prostatic Neoplasms drug therapy   enzymology   metabolism   MeSH
• Nanomedicine MeSH
• Cell Survival drug effects   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

Prostate cancer (PC) is a potentially high-risk disease and the most common cancer in American men. It is a leading cause of cancer-related deaths in men in the US, second only to lung and bronchus cancer. Advanced and metastatic PC is initially treated with androgen deprivation therapy (ADT), but nearly all cases eventually progress to castrate-resistant prostate cancer (CRPC). CRPC is incurable in the metastatic stage but can be slowed by some conventional chemotherapeutics and second-generation ADT, such as enzalutamide and abiraterone. Therefore, novel therapeutic strategies are urgently needed. Prostate-specific membrane antigen (PSMA) is overexpressed in almost all aggressive PCs. PSMA is widely used as a target for PC imaging and drug delivery. Anti-PSMA monoclonal antibodies (mAbs) have been developed as bioligands for diagnostic imaging and targeted PC therapy. However, these mAbs are successfully used in PC imaging and only a few have gone beyond phase-I for targeted therapy. The 5D3 mAb is a novel, high-affinity, and fast-internalizing anti-PSMA antibody. Importantly, 5D3 mAb demonstrates a unique pattern of cellular localization to the centrosome after internalization in PSMA(+) PC3-PIP cells. These characteristics make 5D3 mAb an ideal bioligand to deliver tubulin inhibitors, such as mertansine, to the cell centrosome, leading to mitotic arrest and elimination of dividing PC cells. We have successfully developed a 5D3 mAb- and mertansine (DM1)-based antibody-drug conjugate (ADC) and evaluated it in vitro for binding affinity, internalization, and cytotoxicity. The in vivo therapeutic efficacy of 5D3-DM1 ADC was evaluated in PSMA(+) PC3-PIP and PSMA(-) PC3-Flu mouse models of human PC. This therapeutic study has revealed that this new anti-PSMA ADC can successfully control the growth of PSMA(+) tumors without inducing systemic toxicity.

• MeSH
• Androstenes pharmacology   MeSH
• Androgen Antagonists pharmacology   MeSH
• Antigens, Surface metabolism   MeSH
• Benzamides pharmacology   MeSH
• PC-3 Cells MeSH
• Centrosome metabolism   MeSH
• Phenylthiohydantoin pharmacology   MeSH
• Glutamate Carboxypeptidase II metabolism   MeSH
• Immunoconjugates pharmacology   MeSH
• Humans MeSH
• Tubulin Modulators pharmacology   MeSH
• Antibodies, Monoclonal pharmacology   MeSH
• Mice, Nude MeSH
• Mice MeSH
• Cell Line, Tumor MeSH
• Prostatic Neoplasms, Castration-Resistant drug therapy   metabolism   MeSH
• Nitriles pharmacology   MeSH
• Xenograft Model Antitumor Assays MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

We have synthesized a series of 2-phenyl-3-hydroxy-4(1H)-quinolinone derivatives substituted with one or more fluorine atoms on the quinolone backbone as well as on phenyl ring. The derivatives bearing more fluorine atoms were subjected to modification by nucleophilic substitutions by thiophenol, morpholine, and piperazine derivative. We have tested the prepared compounds in cytotoxic activity assay against cancer cell lines. Four derivatives exhibited micromolar values of IC50 against some of the cancer cell lines, and we have subjected them to cell cycle analysis on CCRF-CEM. Moreover, most active 7-fluoro-3-hydroxy-2-phenyl-6-(phenylthio)quinolin-4(1H)-one inhibits mitosis progression. Cell cycle analysis, in vitro tubulin polymerization assay, and tubulin imaging in cells indicated that the anticancer activity of thiophenol derivative is associated with its ability to inhibit microtubule formation.

• MeSH
• Quinolones chemical synthesis   chemistry   pharmacology   MeSH
• Halogenation MeSH
• HCT116 Cells MeSH
• Humans MeSH
• Tubulin Modulators chemical synthesis   chemistry   pharmacology   MeSH
• Molecular Structure MeSH
• Polymerization drug effects   MeSH
• Tubulin metabolism   MeSH
• Dose-Response Relationship, Drug MeSH
• Structure-Activity Relationship MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Combretastatin A-4 (CA-4) is a highly cytotoxic natural product and several derivatives have been prepared which underwent clinical trial. These investigations revealed that the cis-stilbene moiety of the natural product is prone to undergo cis/trans isomerization under physiological conditions, reducing the overall activity of the drug candidates. Herein, we report the preparation of cis-restrained carbocyclic analogs of CA-4. The compounds, which differ by the size and hybridization of the carbocyclic ring have been evaluated for their cytotoxic properties and their ability to inhibit tubulin polymerization. Biological data, supported by molecular docking studies, identified cyclobutenyl and cyclobutyl derivatives of the natural product as highly promising drug candidates.

• MeSH
• G2 Phase Cell Cycle Checkpoints drug effects   MeSH
• Humans MeSH
• Tubulin Modulators chemical synthesis   metabolism   pharmacology   MeSH
• Molecular Structure MeSH
• Cell Line, Tumor MeSH
• Antineoplastic Agents chemical synthesis   metabolism   pharmacology   MeSH
• Drug Screening Assays, Antitumor MeSH
• Molecular Docking Simulation MeSH
• Stilbenes chemical synthesis   metabolism   pharmacology   MeSH
• Tubulin metabolism   MeSH
• Protein Binding MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The substitution inert platinum agent [Pt(1 S,2 S-diaminocyclohexane)(5,6-dimethyl-1,10-phenanthroline)]2+ (56MeSS, 5) is a potent cytotoxic metallodrug. In contrast to conventional cisplatin or oxaliplatin, the mechanism of action (MoA) of 5 is fundamentally different. However, details of the mechanism by which the 5,6-dimethyl-1,10-phenanthroline ligand contributes to the cytotoxicity of 5 and its derivatives have not been sufficiently clarified so far. Here, we show that 5 and its Pt(IV) derivatives exhibit an intriguing potency in the triple-negative breast cancer cells MDA-MB-231. Moreover, we show that the Pt(IV) derivatives of 5 act by multimodal MoA resulting in the global biological effects, that is, they damage nuclear DNA, reduce the mitochondrial membrane potential, induce the epigenetic processes, and last but not least, the data provide evidence that changes in the organization of cytoskeleton networks are functionally important for 5 and its derivatives, in contrast to clinically used platinum cytostatics, to kill cancer cells.

• MeSH
• Actins antagonists & inhibitors   MeSH
• CHO Cells MeSH
• Cricetulus MeSH
• Cytoskeleton drug effects   MeSH
• DNA, Neoplasm drug effects   MeSH
• Epigenesis, Genetic drug effects   MeSH
• Cricetinae MeSH
• Humans MeSH
• Ligands MeSH
• Microtubules drug effects   MeSH
• Tubulin Modulators pharmacology   MeSH
• Cell Line, Tumor MeSH
• Organoplatinum Compounds chemical synthesis   metabolism   pharmacology   MeSH
• Antineoplastic Agents chemical synthesis   metabolism   pharmacology   MeSH
• Drug Screening Assays, Antitumor MeSH
• Triple Negative Breast Neoplasms drug therapy   MeSH
• Animals MeSH
• Check Tag
• Cricetinae MeSH
• Humans MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Vinorelbin je semisyntetický vinca alkaloid druhé generace, který má nízkou afinitu k neurotubulům, z čehož vyplývá jeho nižší neurotoxicita. Vinca alkaloidy jsou mitotické jedy, které znemožňují správné vytváření mikrotubulů vazbou na tubulin. Způsobují tak depolymerizaci mikrotubulů a rozpouštění mitotického vřeténka, čímž zablokují dělení buněk a zapříčiňují poškození dalších dějů v závislosti na správné funkci mikrotubulů. V onkologii má vinorelbin širší využití, zejména v léčbě u metastazujícího karcinomu prsu a u nemalobuněčného plicního karcinomu, kde je použitelný ve všech stadiích v neoadjuvantní, adjuvantní i paliativní léčbě.

Vinorelbine is a second generation semisynthetic vinca alkaloid that has a low affinity for neurotubules, resulting in lower neurotoxicity. Vinca alkaloids are mitotic poisons that prevent the proper formation of microtubules by binding to tubulin. They cause depolymerization of microtubules and dissolution of the mitotic spindle, thereby blocking cell division and causing damage to other processes depending on the proper functioning of the microtubules. In oncology vinorelbine has a wider use, especially in metastatic breast cancer and non-small cell lung cancer, where it is applicable at all stages of non-adjuvant and palliative treatment.

• MeSH
• Adult MeSH
• Fatal Outcome MeSH
• Antineoplastic Agents, Phytogenic therapeutic use   MeSH
• Remission Induction MeSH
• Humans MeSH
• Young Adult MeSH
• Tubulin Modulators therapeutic use   MeSH
• Carcinoma, Non-Small-Cell Lung * diagnosis   therapy   MeSH
• Disease Progression MeSH
• Vinorelbine * adverse effects   therapeutic use   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Female MeSH
• Publication type
• Case Reports MeSH

Microtubule dynamics is one of the major targets for new chemotherapeutic agents. This communication presents the synthesis and biological profiling of steroidal dimers based on estradiol, testosterone and pregnenolone bridged by 2,6-bis(azidomethyl)pyridine between D rings. The biological profiling revealed unique properties of the estradiol dimer including cytotoxic activities on a panel of 11 human cell lines, ability to arrest in the G2/M phase of the cell cycle accompanied with the attenuation of DNA/RNA synthesis. Thorough investigation precluded a genomic mechanism of action and revealed that the estradiol dimer acts at the cytoskeletal level by inhibiting tubulin polymerization. Further studies showed that estradiol dimer, but none of the other structurally related dimeric steroids, inhibited assembly of purified tubulin (IC50, 3.6 μM). The estradiol dimer was more potent than 2-methoxyestradiol, an endogenous metabolite of 17β-estradiol and well-studied microtubule polymerization inhibitor with antitumor effects that was evaluated in clinical trials. Further, it was equipotent to nocodazole (IC50, 1.5 μM), an antimitotic small molecule of natural origin. Both estradiol dimer and nocodazole completely and reversibly depolymerized microtubules in interphase U2OS cells at 2.5 μM concentration. At lower concentrations (50 nM), estradiol dimer decreased the microtubule dynamics and growth life-time and produced comparable effect to nocodazole on the microtubule dynamicity. In silico modeling predicted that estradiol dimer binds to the colchicine-binding site in the tubulin dimer. Finally, dimerization of the steroids abolished their ability to induce transactivation by estrogen receptor α and androgen receptors. Although other steroids were reported to interact with microtubules, the estradiol dimer represents a new structural type of steroid inhibitor of tubulin polymerization and microtubule dynamics, bearing antimitotic and cytotoxic activity in cancer cell lines.

• MeSH
• Cell Cycle MeSH
• Estradiol chemistry   pharmacology   MeSH
• Estrogens chemistry   pharmacology   MeSH
• Humans MeSH
• Microtubules drug effects   physiology   MeSH
• Tubulin Modulators chemistry   pharmacology   MeSH
• Tumor Cells, Cultured MeSH
• Neoplasms drug therapy   metabolism   pathology   MeSH
• Polymerization MeSH
• Cell Proliferation MeSH
• Tubulin chemistry   drug effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH