6-Nitrobenzo[b]thiophene 1,1-dioxide (Stattic) is a potent signal transducer and activator of the transcription 3 (STAT3) inhibitor developed originally for anticancer therapy. However, Stattic harbors several STAT3 inhibition-independent biological effects. To improve the properties of Stattic, we prepared a series of analogues derived from 6-aminobenzo[b]thiophene 1,1-dioxide, a compound directly obtained from the reduction of Stattic, that includes a methoxybenzylamino derivative (K2071) with optimized physicochemical characteristics, including the ability to cross the blood-brain barrier. Besides inhibiting the interleukin-6-stimulated activity of STAT3 mediated by tyrosine 705 phosphorylation, K2071 also showed cytotoxicity against a set of human glioblastoma-derived cell lines. In contrast to the core compound, a part of K2071 cytotoxicity reflected a STAT3 inhibition-independent block of mitotic progression in the prophase, affecting mitotic spindle formation, indicating that K2071 also acts as a mitotic poison. Compared to Stattic, K2071 was significantly less thiol-reactive. In addition, K2071 affected cell migration, suppressed cell proliferation in tumor spheroids, exerted cytotoxicity for glioblastoma temozolomide-induced senescent cells, and inhibited the secretion of the proinflammatory cytokine monocyte chemoattractant protein 1 (MCP-1) in senescent cells. Importantly, K2071 was well tolerated in mice, lacking manifestations of acute toxicity. The structure-activity relationship analysis of the K2071 molecule revealed the necessity of the para-substituted methoxyphenyl motif for antimitotic but not overall cytotoxic activity of its derivatives. Altogether, these results indicate that compound K2071 is a novel Stattic-derived STAT3 inhibitor and a mitotic poison with anticancer and senotherapeutic properties that is effective on glioblastoma cells and may be further developed as an agent for glioblastoma therapy.

BIOCEV 1st Faculty of Medicine Charles University Prumyslova 595 Vestec 252 50 Czech Republic

Biomedical Research Centre University Hospital Hradec Kralove Sokolska 581 Hradec Kralove 500 05 Czech Republic

CZ OPENSCREEN Institute of Molecular Genetics of the Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic

Danish Cancer Institute Strandboulevarden 49 DK 2100 Copenhagen Denmark

Division of Genome Biology Department of Medical Biochemistry and Biophysics Science for Life Laboratory Karolinska Institutet Stockholm 171 77 Sweden

Faculty of Science Department of Chemistry University of Hradec Kralove Rokitanskeho 62 Hradec Kralove 500 03 Czech Republic

Laboratory of Biology of Cytoskeleton Institute of Molecular Genetics of the Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic

Laboratory of Genome Integrity Institute of Molecular Genetics of the Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic

Laboratory of Immunological and Tumour Models Institute of Molecular Genetics of the Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic

Zobrazit více v PubMed

Gai C.; Harnor S. J.; Zhang S.; Cano C.; Zhuang C.; Zhao Q. Advanced approaches of developing targeted covalent drugs. RSC Med. Chem. 2022, 13 (12), 1460–1475. 10.1039/d2md00216g. PubMed DOI PMC

Jackson P. A.; Widen J. C.; Harki D. A.; Brummond K. M. Covalent Modifiers: A Chemical Perspective on the Reactivity of α,β-Unsaturated Carbonyls with Thiols via Hetero-Michael Addition Reactions. J. Med. Chem. 2017, 60 (3), 839–885. 10.1021/acs.jmedchem.6b00788. PubMed DOI PMC

Huang F.; Han X.; Xiao X.; Zhou J. Covalent Warheads Targeting Cysteine Residue: The Promising Approach in Drug Development. Molecules 2022, 27 (22), 7728.10.3390/molecules27227728. PubMed DOI PMC

Schust J.; Sperl B.; Hollis A.; Mayer T. U.; Berg T. S. Stattic: A Small-Molecule Inhibitor of STAT3 Activation and Dimerization. Chem. Biol. 2006, 13 (11), 1235–1242. 10.1016/j.chembiol.2006.09.018. PubMed DOI

Poria D. K.; Sheshadri N.; Balamurugan K.; Sharan S.; Sterneck E. The STAT3 inhibitor Stattic acts independently of STAT3 to decrease histone acetylation and modulate gene expression. J. Biol. Chem. 2021, 296, 100220.10.1074/jbc.RA120.016645. PubMed DOI PMC

Uchihara Y.; Ohe T.; Mashino T.; Kidokoro T.; Tago K.; Tamura H.; Funakoshi-Tago M. N-Acetyl cysteine prevents activities of STAT3 inhibitors, Stattic and BP-1–102 independently of its antioxidant properties. Pharmacol. Rep. 2019, 71 (6), 1067–1078. 10.1016/j.pharep.2019.05.021. PubMed DOI

Hanahan D.; Weinberg R. A. Hallmarks of cancer: the next generation. Cell 2011, 144 (5), 646–674. 10.1016/j.cell.2011.02.013. PubMed DOI

Verhoeven Y.; Tilborghs S.; Jacobs J.; De Waele J.; Quatannens D.; Deben C.; Prenen H.; Pauwels P.; Trinh X. B.; Wouters A.; et al. The potential and controversy of targeting STAT family members in cancer. Semin. Cancer Biol. 2020, 60, 41–56. 10.1016/j.semcancer.2019.10.002. PubMed DOI

Bromberg J. F. Activation of STAT proteins and growth control. Bioessays 2001, 23 (2), 161–169. 10.1002/1521-1878(200102)23:2<161::AID-BIES1023>3.0.CO;2-0. PubMed DOI

Yu H.; Jove R. The STATs of cancer--new molecular targets come of age. Nat. Rev. Cancer 2004, 4 (2), 97–105. 10.1038/nrc1275. PubMed DOI

Bromberg J. F.; Wrzeszczynska M. H.; Devgan G.; Zhao Y.; Pestell R. G.; Albanese C.; Darnell J. E. Jr. Stat3 as an oncogene. Cell 1999, 98 (3), 295–303. 10.1016/s0092-8674(00)81959-5. PubMed DOI

Wang H.-Q.; Man Q.-W.; Huo F.-Y.; Gao X.; Lin H.; Li S.-R.; Wang J.; Su F.-C.; Cai L.; Shi Y.; et al. STAT3 pathway in cancers: Past, present, and future. MedComm 2022, 3 (2), e12410.1002/mco2.124. PubMed DOI PMC

Hu Y.; Dong Z.; Liu K. Unraveling the complexity of STAT3 in cancer: molecular understanding and drug discovery. J. Exp. Clin. Cancer Res. 2024, 43 (1), 23.10.1186/s13046-024-02949-5. PubMed DOI PMC

Chen X.; Yang W.; Deng X.; Ye S.; Xiao W. Interleukin-6 promotes proliferative vitreoretinopathy by inducing epithelial-mesenchymal transition via the JAK1/STAT3 signaling pathway. Mol. Vis. 2020, 26, 517–529. PubMed PMC

Steinman R. A.; Robinson A. R.; Feghali-Bostwick C. A. Antifibrotic effects of roscovitine in normal and scleroderma fibroblasts. PLoS One 2012, 7 (11), e4856010.1371/journal.pone.0048560. PubMed DOI PMC

Liebl M. C.; Hofmann T. G. Regulating the p53 Tumor Suppressor Network at PML Biomolecular Condensates. Cancers 2022, 14 (19), 4549.10.3390/cancers14194549. PubMed DOI PMC

Vultur A.; Cao J.; Arulanandam R.; Turkson J.; Jove R.; Greer P.; Craig A.; Elliott B.; Raptis L. Cell-to-cell adhesion modulates Stat3 activity in normal and breast carcinoma cells. Oncogene 2004, 23 (15), 2600–2616. 10.1038/sj.onc.1207378. PubMed DOI

von Manstein V.; Groner B. Tumor cell resistance against targeted therapeutics: the density of cultured glioma tumor cells enhances Stat3 activity and offers protection against the tyrosine kinase inhibitor canertinib. MedChemComm 2017, 8 (1), 96–102. 10.1039/C6MD00463F. PubMed DOI PMC

Di L.; Kerns E. H.; Fan K.; McConnell O. J.; Carter G. T. High throughput artificial membrane permeability assay for blood-brain barrier. Eur. J. Med. Chem. 2003, 38 (3), 223–232. 10.1016/s0223-5234(03)00012-6. PubMed DOI

Park M. C.; Jeong H.; Son S. H.; Kim Y.; Han D.; Goughnour P. C.; Kang T.; Kwon N. H.; Moon H. E.; Paek S. H.; et al. Novel Morphologic and Genetic Analysis of Cancer Cells in a 3D Microenvironment Identifies STAT3 as a Regulator of Tumor Permeability Barrier Function. Cancer Res. 2016, 76 (5), 1044–1054. 10.1158/0008-5472.CAN-14-2611. PubMed DOI

Clark J.; Edwards S.; Feber A.; Flohr P.; John M.; Giddings I.; Crossland S.; Stratton M. R.; Wooster R.; Campbell C.; et al. Genome-wide screening for complete genetic loss in prostate cancer by comparative hybridization onto cDNA microarrays. Oncogene 2003, 22 (8), 1247–1252. 10.1038/sj.onc.1206247. PubMed DOI

Seim I.; Jeffery P. L.; Thomas P. B.; Nelson C. C.; Chopin L. K. Whole-Genome Sequence of the Metastatic PC3 and LNCaP Human Prostate Cancer Cell Lines. G3 2017, 7 (6), 1731–1741. 10.1534/g3.117.039909. PubMed DOI PMC

Buettner R.; Corzano R.; Rashid R.; Lin J.; Senthil M.; Hedvat M.; Schroeder A.; Mao A.; Herrmann A.; Yim J.; et al. Alkylation of cysteine 468 in Stat3 defines a novel site for therapeutic development. ACS Chem. Biol. 2011, 6 (5), 432–443. 10.1021/cb100253e. PubMed DOI PMC

Heidelberger S.; Zinzalla G.; Antonow D.; Essex S.; Piku Basu B.; Palmer J.; Husby J.; Jackson P. J.; Rahman K. M.; Wilderspin A. F.; et al. Investigation of the protein alkylation sites of the STAT3:STAT3 inhibitor Stattic by mass spectrometry. Bioorg. Med. Chem. Lett. 2013, 23 (16), 4719–4722. 10.1016/j.bmcl.2013.05.066. PubMed DOI

Elmaci I. ˙.; Altinoz M. A.; Sari R.; Bolukbasi F. H. Phosphorylated Histone H3 (PHH3) as a Novel Cell Proliferation Marker and Prognosticator for Meningeal Tumors: A Short Review. Appl. Immunohistochem. Mol. Morphol. 2018, 26 (9), 627–631. 10.1097/PAI.0000000000000499. PubMed DOI

Park C. H.; Kim K. T. Apoptotic phosphorylation of histone H3 on Ser-10 by protein kinase Cδ. PLoS One 2012, 7 (9), e4430710.1371/journal.pone.0044307. PubMed DOI PMC

Dráberová E.; Sulimenko V.; Sulimenko T.; Böhm K. J.; Dráber P. Recovery of tubulin functions after freeze-drying in the presence of trehalose. Anal. Biochem. 2010, 397 (1), 67–72. 10.1016/j.ab.2009.10.016. PubMed DOI

Teng Y.; Ross J. L.; Cowell J. K. The involvement of JAK-STAT3 in cell motility, invasion, and metastasis. JAK-STAT 2014, 3 (1), e2808610.4161/jkst.28086. PubMed DOI PMC

Chrienova Z.; Rysanek D.; Oleksak P.; Stary D.; Bajda M.; Reinis M.; Mikyskova R.; Novotny O.; Andrys R.; Skarka A.; et al. Discovery of small molecule mechanistic target of rapamycin inhibitors as anti-aging and anti-cancer therapeutics. Front. Aging Neurosci. 2022, 14, 1048260.10.3389/fnagi.2022.1048260. PubMed DOI PMC

Coppe J. P.; Patil C. K.; Rodier F.; Sun Y.; Munoz D. P.; Goldstein J.; Nelson P. S.; Desprez P. Y.; Campisi J. Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol. 2008, 6 (12), e30110.1371/journal.pbio.0060301. PubMed DOI PMC

Rayan A.; Raiyn J.; Falah M. Nature is the best source of anticancer drugs: Indexing natural products for their anticancer bioactivity. PLoS One 2017, 12 (11), e018792510.1371/journal.pone.0187925. PubMed DOI PMC

Cui W.; Aouidate A.; Wang S.; Yu Q.; Li Y.; Yuan S. Discovering Anti-Cancer Drugs via Computational Methods. Front. Pharmacol. 2020, 11, 733.10.3389/fphar.2020.00733. PubMed DOI PMC

Pathania S.; Narang R. K.; Rawal R. K. Role of sulphur-heterocycles in medicinal chemistry: An update. Eur. J. Med. Chem. 2019, 180, 486–508. 10.1016/j.ejmech.2019.07.043. PubMed DOI

Mishra R.; Kumar N.; Mishra I.; Sachan N. A Review on Anticancer Activities of Thiophene and Its Analogs. Mini-Rev. Med. Chem. 2020, 20 (19), 1944–1965. 10.2174/1389557520666200715104555. PubMed DOI

Xiang J.; Zhang Z.; Mu Y.; Xu X.; Guo S.; Liu Y.; Russo D. P.; Zhu H.; Yan B.; Bai X. Discovery of Novel Tricyclic Thiazepine Derivatives as Anti-Drug-Resistant Cancer Agents by Combining Diversity-Oriented Synthesis and Converging Screening Approach. ACS Comb. Sci. 2016, 18 (5), 230–235. 10.1021/acscombsci.6b00010. PubMed DOI PMC

Mishra R.; Sharma P. K.; Verma P. K.; Tomer I.; Mathur G.; Dhakad P. K. Biological Potential of Thiazole Derivatives of Synthetic Origin. J. Heterocycl. Chem. 2017, 54 (4), 2103–2116. 10.1002/jhet.2827. DOI

Taghour M. S.; Elkady H.; Eldehna W. M.; El-Deeb N. M.; Kenawy A. M.; Elkaeed E. B.; Alsfouk A. A.; Alesawy M. S.; Metwaly A. M.; Eissa I. H. Design and synthesis of thiazolidine-2,4-diones hybrids with 1,2-dihydroquinolones and 2-oxindoles as potential VEGFR-2 inhibitors: in-vitro anticancer evaluation and in-silico studies. J. Enzym. Inhib. Med. Chem. 2022, 37 (1), 1903–1917. 10.1080/14756366.2022.2085693. PubMed DOI PMC

Hekal M. H.; Farag P. S.; Hemdan M. M.; El-Sayed A. A.; Hassaballah A. I.; El-Sayed W. M. New 1,3,4-thiadiazoles as potential anticancer agents: pro-apoptotic, cell cycle arrest, molecular modelling, and ADMET profile. RSC Adv. 2023, 13 (23), 15810–15825. 10.1039/d3ra02716c. PubMed DOI PMC

Mishra R.; Kumar N.; Sachan N. Synthesis, Biological Evaluation, and Docking Analysis of Novel Tetrahydrobenzothiophene Derivatives. Lett. Drug Des. Discov. 2022, 19 (6), 530–540. 10.2174/1570180819666220117123958. DOI

Boyd D. R.; Sharma N. D.; McMurray B.; Haughey S. A.; Allen C. C.; Hamilton J. T.; McRoberts W. C.; More O’Ferrall R. A.; Nikodinovic-Runic J.; Coulombel L. A.; et al. Bacterial dioxygenase- and monooxygenase-catalysed sulfoxidation of benzo[b]thiophenes. Org. Biomol. Chem. 2012, 10 (4), 782–790. 10.1039/c1ob06678a. PubMed DOI

Du G.; Du W.; An Y.; Wang M.; Hao F.; Tong X.; Gong Q.; He X.; Jiang H.; He W.; et al. Design, synthesis, and LFA-1/ICAM-1 antagonist activity evaluation of Lifitegrast analogues. Med. Chem. Res. 2022, 31 (4), 555–579. 10.1007/s00044-022-02851-9. PubMed DOI PMC

Villar R.; Encio I.; Migliaccio M.; Gil M. J.; Martinez-Merino V. Synthesis and cytotoxic activity of lipophilic sulphonamide derivatives of the benzo[b]thiophene 1,1-dioxide. Bioorg. Med. Chem. 2004, 12 (5), 963–968. 10.1016/j.bmc.2003.12.012. PubMed DOI

Serby M. D.; Zhao H.; Szczepankiewicz B. G.; Kosogof C.; Xin Z.; Liu B.; Liu M.; Nelson L. T.; Kaszubska W.; Falls H. D.; et al. 2,4-diaminopyrimidine derivatives as potent growth hormone secretagogue receptor antagonists. J. Med. Chem. 2006, 49 (8), 2568–2578. 10.1021/jm0510934. PubMed DOI

Madec D.; Mingoia F.; Macovei C.; Maitro G.; Giambastiani G.; Poli G. New Enantiopure Bis(thioether) and Bis(sulfoxide) Ligands from Benzothiophene. Eur. J. Org Chem. 2005, 2005 (3), 552–557. 10.1002/ejoc.200400547. DOI

Lawson C.; Ahmed Alta T. B.; Moschou G.; Skamnaki V.; Solovou T. G. A.; Topham C.; Hayes J.; Snape T. J. Novel diarylamides and diarylureas with N-substitution dependent activity against medulloblastoma. Eur. J. Med. Chem. 2021, 225, 113751.10.1016/j.ejmech.2021.113751. PubMed DOI

Banks W. A. Drug delivery to the brain in Alzheimer’s disease: consideration of the blood-brain barrier. Adv. Drug Deliv. Rev. 2012, 64 (7), 629–639. 10.1016/j.addr.2011.12.005. PubMed DOI PMC

Harley M. E.; Allan L. A.; Sanderson H. S.; Clarke P. R. Phosphorylation of Mcl-1 by CDK1-cyclin B1 initiates its Cdc20-dependent destruction during mitotic arrest. EMBO J. 2010, 29 (14), 2407–2420. 10.1038/emboj.2010.112. PubMed DOI PMC

Takenaka K.; Moriguchi T.; Nishida E. Activation of the protein kinase p38 in the spindle assembly checkpoint and mitotic arrest. Science 1998, 280 (5363), 599–602. 10.1126/science.280.5363.599. PubMed DOI

Xu N.; Hegarat N.; Black E. J.; Scott M. T.; Hochegger H.; Gillespie D. A. Akt/PKB suppresses DNA damage processing and checkpoint activation in late G2. J. Cell Biol. 2010, 190 (3), 297–305. 10.1083/jcb.201003004. PubMed DOI PMC

Ng D. C.; Lin B. H.; Lim C. P.; Huang G.; Zhang T.; Poli V.; Cao X. Stat3 regulates microtubules by antagonizing the depolymerization activity of stathmin. J. Cell Biol. 2006, 172 (2), 245–257. 10.1083/jcb.200503021. PubMed DOI PMC

Morris E. J.; Kawamura E.; Gillespie J. A.; Balgi A.; Kannan N.; Muller W. J.; Roberge M.; Dedhar S. Stat3 regulates centrosome clustering in cancer cells via Stathmin/PLK1. Nat. Commun. 2017, 8, 15289.10.1038/ncomms15289. PubMed DOI PMC

Podolak M.; Holota S.; Deyak Y.; Dziduch K.; Dudchak R.; Wujec M.; Bielawski K.; Lesyk R.; Bielawska A. Tubulin inhibitors. Selected scaffolds and main trends in the design of novel anticancer and antiparasitic agents. Bioorg. Chem. 2024, 143, 107076.10.1016/j.bioorg.2023.107076. PubMed DOI

Walker S. R.; Chaudhury M.; Nelson E. A.; Frank D. A. Microtubule-targeted chemotherapeutic agents inhibit signal transducer and activator of transcription 3 (STAT3) signaling. Mol. Pharmacol. 2010, 78 (5), 903–908. 10.1124/mol.110.066316. PubMed DOI

Huang H. L.; Chao M. W.; Chen C. C.; Cheng C. C.; Chen M. C.; Lin C. F.; Liou J. P.; Teng C. M.; Pan S. L. LTP-1, a novel antimitotic agent and Stat3 inhibitor, inhibits human pancreatic carcinomas in vitro and in vivo. Sci. Rep. 2016, 6, 27794.10.1038/srep27794. PubMed DOI PMC

Sulimenko V.; Dráberová E.; Dráber P. γ-Tubulin in microtubule nucleation and beyond. Front. Cell Dev. Biol. 2022, 10, 880761.10.3389/fcell.2022.880761. PubMed DOI PMC

Huang W.; Dong Z.; Wang F.; Peng H.; Liu J. Y.; Zhang J. T. A small molecule compound targeting STAT3 DNA-binding domain inhibits cancer cell proliferation, migration, and invasion. ACS Chem. Biol. 2014, 9 (5), 1188–1196. 10.1021/cb500071v. PubMed DOI PMC

Li Z.; Zhu T.; Xu Y.; Wu C.; Chen J.; Ren Y.; Kong L.; Sun S.; Guo W.; Wang Y.; et al. A novel STAT3 inhibitor, HJC0152, exerts potent antitumor activity in glioblastoma. Am. J. Cancer Res. 2019, 9 (4), 699–713. PubMed PMC

Chang Y. C.; Nalbant P.; Birkenfeld J.; Chang Z. F.; Bokoch G. M. GEF-H1 couples nocodazole-induced microtubule disassembly to cell contractility via RhoA. Mol. Biol. Cell 2008, 19 (5), 2147–2153. 10.1091/mbc.e07-12-1269. PubMed DOI PMC

Takesono A.; Heasman S. J.; Wojciak-Stothard B.; Garg R.; Ridley A. J. Microtubules regulate migratory polarity through Rho/ROCK signaling in T cells. PLoS One 2010, 5 (1), e877410.1371/journal.pone.0008774. PubMed DOI PMC

Kuilman T.; Michaloglou C.; Vredeveld L. C.; Douma S.; van Doorn R.; Desmet C. J.; Aarden L. A.; Mooi W. J.; Peeper D. S. Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell 2008, 133 (6), 1019–1031. 10.1016/j.cell.2008.03.039. PubMed DOI

Li R.; Dong J.; Bu X. Q.; Huang Y.; Yang J. Y.; Dong X.; Liu J. Retracted: Interleukin-6 promotes the migration and cellular senescence and inhibits apoptosis of human intrahepatic biliary epithelial cells. J. Cell. Biochem. 2018, 119 (2), 2135–2143. 10.1002/jcb.26375. PubMed DOI

Yun U. J.; Park S. E.; Jo Y. S.; Kim J.; Shin D. Y. DNA damage induces the IL-6/STAT3 signaling pathway, which has anti-senescence and growth-promoting functions in human tumors. Cancer Lett. 2012, 323 (2), 155–160. 10.1016/j.canlet.2012.04.003. PubMed DOI

Kojima H.; Inoue T.; Kunimoto H.; Nakajima K. IL-6-STAT3 signaling and premature senescence. JAK-STAT 2013, 2 (4), e2576310.4161/jkst.25763. PubMed DOI PMC

Wang L.; Lan J.; Tang J.; Luo N. MCP-1 targeting: Shutting off an engine for tumor development (Review). Oncol. Lett. 2021, 23 (1), 26.10.3892/ol.2021.13144. PubMed DOI PMC

Nováková M.; Dráberová E.; Schürmann W.; Czihak G.; Viklický V.; Dr-aber P. γ-tubulin redistribution in taxol-treated mitotic cells probed by monoclonal antibodies. Cell Motil Cytoskeleton 1996, 33 (1), 38–51. 10.1002/(sici)1097-0169(1996)33:1<38::aid-cm5>3.0.co;2-e. PubMed DOI

Dimri G. P.; Lee X.; Basile G.; Acosta M.; Scott G.; Roskelley C.; Medrano E. E.; Linskens M.; Rubelj I.; Pereira-Smith O.; et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl. Acad. Sci. U.S.A. 1995, 92 (20), 9363–9367. 10.1073/pnas.92.20.9363. PubMed DOI PMC

Müller T.; Sedlák D.; Bartunék P. Laboratory Information Systems to High-Throughput Screening. Chem. Listy 2017, 111 (11), 766–771.

Chrienova Z.; Rysanek D.; Novak J.; Vasicova P.; Oleksak P.; Andrys R.; Skarka A.; Dumanovic J.; Milovanovic Z.; Jacevic V.; et al. Frentizole derivatives with mTOR inhibiting and senomorphic properties. Biomed. Pharmacother. 2023, 167, 115600.10.1016/j.biopha.2023.115600. PubMed DOI

Ershov D.; Phan M.-S.; Pylvänäinen J. W.; Rigaud S. U.; Blanc L. L.; Charles-Orszag A.; Conway J. R. W.; Laine R. F.; Roy N. H.; Bonazzi D.; et al. Bringing TrackMate into the era of machine-learning and deep-learning. bioRxiv 2021, 2021.09.03.458852.10.1101/2021.09.03.458852. DOI

Ershov D.; Phan M. S.; Pylvanainen J. W.; Rigaud S. U.; Le Blanc L.; Charles-Orszag A.; Conway J. R. W.; Laine R. F.; Roy N. H.; Bonazzi D.; et al. TrackMate 7: integrating state-of-the-art segmentation algorithms into tracking pipelines. Nat. Methods 2022, 19 (7), 829–832. 10.1038/s41592-022-01507-1. PubMed DOI

Schindelin J.; Arganda-Carreras I.; Frise E.; Kaynig V.; Longair M.; Pietzsch T.; Preibisch S.; Rueden C.; Saalfeld S.; Schmid B.; et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 2012, 9 (7), 676–682. 10.1038/nmeth.2019. PubMed DOI PMC

Berg S.; Kutra D.; Kroeger T.; Straehle C. N.; Kausler B. X.; Haubold C.; Schiegg M.; Ales J.; Beier T.; Rudy M.; et al. ilastik: interactive machine learning for (bio)image analysis. Nat. Methods 2019, 16 (12), 1226–1232. 10.1038/s41592-019-0582-9. PubMed DOI

Nussbaum-Krammer C. I.; Neto M. F.; Brielmann R. M.; Pedersen J. S.; Morimoto R. I. Investigating the spreading and toxicity of prion-like proteins using the metazoan model organism C. elegans. J. Vis. Exp. 2015, (95), e52321.10.3791/52321-v. PubMed DOI PMC

Gaskin F.; Cantor C. R.; Shelanski M. L. Turbidimetric studies of the in vitro assembly and disassembly of porcine neurotubules. J. Mol. Biol. 1974, 89 (4), 737–755. 10.1016/0022-2836(74)90048-5. PubMed DOI