Dinaciclib

Manufactured by MedChemExpress

Sourced in United Kingdom, United States

Dinaciclib is a cyclin-dependent kinase (CDK) inhibitor that targets CDK1, CDK2, CDK5, and CDK9. It is a laboratory reagent used for research purposes.

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Dinaciclib (HY-10492), (2 citations)

6 protocols using dinaciclib

Preparation and Handling of Chemical Compounds

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PLX51107 was kindly provided by Plexxikon or purchased from MedChemExpress. Dinaciclib, cytarabine, etoposide, IWR-1, BML-284, and CHIR-99,021 (laduviglusib) were obtained from MedChemExpress. All chemical compounds were dissolved in Dimethyl Sulfoxide (DMSO) to prepare a 10 mM stock solution and were stored at -80 C for long-term storage as recommended.

Marr A.R., Halpin M., Corbin D.L., Asemelash Y., Sher S., Gordon B.K., Whipp E.C., Mitchell S., Harrington B.K., Orwick S., Benrashid S., Goettl V.M., Yildiz V., Mitchell A.D., Cahn O., Mims A.S., Larkin K.T., Long M., Blachly J., Woyach J.A., Lapalombella R, & Grieselhuber N.R. (2024). The multi-CDK inhibitor dinaciclib reverses bromo- and extra-terminal domain (BET) inhibitor resistance in acute myeloid leukemia via inhibition of Wnt/β-catenin signaling. Experimental Hematology & Oncology, 13, 27.

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Anticancer Therapeutic Agents Evaluation

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The following drugs were used in the course of the study: RP-6306 (this study), RP-6421 (this study) AZD1775 (Selleckchem, S1525), dinaciclib (MedChemExpress, HY-10492), PF-06873600 (MedChemExpress, HY-114177), RO-3306 (Selleckchem, S7747), gemcitabine (Cayman Chemicals, 9003096) and hydroxyurea (Sigma-Aldrich cat. no. H8627). Synthesis of RP-6306 and RP-6421 is described in the Supplementary Information. Concentration and duration of treatment is indicated in the legends of the corresponding figures.

Gallo D., Young J.T., Fourtounis J., Martino G., Álvarez-Quilón A., Bernier C., Duffy N.M., Papp R., Roulston A., Stocco R., Szychowski J., Veloso A., Alam H., Baruah P.S., Fortin A.B., Bowlan J., Chaudhary N., Desjardins J., Dietrich E., Fournier S., Fugère-Desjardins C., Goullet de Rugy T., Leclaire M.E., Liu B., Bhaskaran V., Mamane Y., Melo H., Nicolas O., Singhania A., Szilard R.K., Tkáč J., Yin S.Y., Morris S.J., Zinda M., Marshall C.G, & Durocher D. (2022). CCNE1 amplification is synthetic lethal with PKMYT1 kinase inhibition. Nature, 604(7907), 749-756.

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Culturing and Treating Cancer Cell Lines

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Cancer cell lines used in this study were grown in Dulbecco’s Modified Eagle’s Medium (DMEM; Genesee Scientific, San Diego, CA, #25-500) (786-O, RCC4, UM-RC2, MDA-MB-231, MCF-7), RPMI 1640 (Thermo Fisher Scientific, Waltham, MA, #A1049101) (TK-10, T-47D), McCoy’s 5A (Genesee Scientific, #25-518) (Caki-1, Caki-2), or Eagle’s Minimum Essential Medium (EMEM; Sigma-Aldrich, St. Louis, MO, #M4655) (MDCK) supplemented with 10% Fetal Bovine Serum (FBS; Genesee Scientific, #25-514) and 1% Penicillin/Streptomycin (P/S; Genesee Scientific, #25-512) in 5% CO₂ at 37°C. RPTEC cells were grown in DMEM/F12 medium with 36 ng/mL hydrocortisone (Thermo Fisher Scientific, #SH30261.01, #AC352450010), 5 μg/mL insulin, 5 μg/mL apo-transferrin, 5 ng/mL sodium selenite (Sigma-Aldrich, #I0516-5ML, #T1147-100 MG, #S5261-10 G), 50 ng/mL human EGF (PeproTech, Cranbury, NJ #AF-100-15), 1% P/S, and 100 μg/mL G418 (Selleck Chemicals, Houston, TX, #S3028). Dinaciclib and Q-VD-OPh (MedChem Express, LLC, Monmouth Junction, NJ, #HY-10492, #HY-12305) were diluted in Dimethyl Sulfoxide (DMSO) and serially diluted for each experiment. ABT-263 (BioVision Inc., Milpitas, CA, #2467-5), Staurosporine (BioVision Inc., #1048-01) were diluted in DMSO and used at concentrations indicated in the manuscript.

Nelson L.J., Castro K.E., Xu B., Li J., Dinh N.B., Thompson J.M., Woytash J., Kipp K.R, & Razorenova O.V. (2022). Synthetic lethality of cyclin-dependent kinase inhibitor Dinaciclib with VHL-deficiency allows for selective targeting of clear cell renal cell carcinoma. Cell Cycle, 21(10), 1103-1119.

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Characterization of ICEC-Series Compounds

Cited 2 times

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ATPγS was purchased from Sigma Aldrich; THZ1 was purchased from EMD Millipore; LDC4297 and dinaciclib were purchased from MedChemExpress (distributed by Insight Biotechnology, Wembley, UK). ICEC-series pyrazolopyrimidine compounds were synthesized and characterized during the course of the drug discovery program that led to the discovery of ICEC0942^{5 (link),30 (link)–33 ,41}. Further characterization data for ICEC0510-R and S enantiomers, ICEC0768, ICEC0880, ICEC0914, and CT7030 are provided above. After retrieval from cold storage, integrity of ICEC-series compounds was verified using UV-vis spectroscopy and liquid chromatography/mass spectrometry (see below for detailed methods). The results of this characterization are summarized in Supplementary Table 12.

Cushing V.I., Koh A.F., Feng J., Jurgaityte K., Bondke A., Kroll S.H., Barbazanges M., Scheiper B., Bahl A.K., Barrett A.G., Ali S., Kotecha A, & Greber B.J. (2024). High-resolution cryo-EM of the human CDK-activating kinase for structure-based drug design. Nature Communications, 15, 2265.

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Culturing and Treating CRC Cell Lines

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Human CRC cell lines, RKO, HT29, SW620, SW1116, SW480, and HCT116 were purchased from ATCC (Rockville, MD, USA). Human CRC cell lines LOVO and DLD-1 were obtained from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences. CRC cell lines RKO, HT29, SW620, SW1116, SW480, HCT116, and DLD-1 were cultured in high-glucose RPMI-1640 supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37 °C with 5% CO₂. LOVO was cultured in high-glucose DMEM supplemented as described above. Palbociclib, a highly selective CDK4/6 inhibitor, was purchased from SELLECK (Cat. No. S1579, USA). Dinaciclib, an effective selective CDK inhibitor that inhibits CDK2, CDK5, CDK1, and CDK9, was purchased from MedChemExpress (Cat. No. HY-10492, USA).

Peng Y., Feng H., Wang C., Song Z., Zhang Y., Liu K., Cheng X, & Zhao R. (2021). The role of E26 transformation-specific variant transcription factor 5 in colorectal cancer cell proliferation and cell cycle progression. Cell Death & Disease, 12(5), 427.

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Comprehensive Chemical Inhibitor Sourcing

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Dinaciclib was purchased from MedChem Express (Monmouth Junction, NJ, USA), and CDK2/9 inhibitor (iCDK2/9) (CAS No. 507487–89-0), Purvalanol A, and SNS032 were purchased from Merck Millipore (Merck KGaA, Darmstadt, Germany). Atuveciclib and LDC000067 were purchased from Selleck (Houston, TX, USA). The BRAF V600E inhibitor vemurafenib and MEK inhibitor trametinib were purchased from LC Laboratories (Woburn, MA, USA). For the in vitro experiments, all of the chemical inhibitors were formulated in DMSO and the final concentration of DMSO was < 0.1%.

Xu X., Eshima S., Kato S., Fisher D.E., Sakurai H., Hayakawa Y, & Yokoyama S. (2019). Rational combination therapy for melanoma with dinaciclib by targeting BAK-dependent cell death. Molecular cancer therapeutics, 19(2), 627-636.

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