Irinotecan hydrochloride

Manufactured by Merck Group

Sourced in United States

Irinotecan hydrochloride is a topoisomerase I inhibitor used as a laboratory reagent. It functions by interfering with the action of topoisomerase I, an enzyme involved in DNA replication and transcription.

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Lab products found in correlation

22 protocols using irinotecan hydrochloride

Carbodiimide-mediated Bioconjugation Protocol

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Urea (99%), citric acid (99.5%), N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC) (99%), N-hydroxysuccinimide (NHS) (98%), ethanol (99.9%), phosphate buffered saline (PBS) pH 7.4, anhydrous N,N-dimethylformamide (DMF), rhodamine (99.5%), copper (II) sulfate (99%), ascorbic acid (99.5%), azide-PEG300-biotin, irinotecan hydrochloride (IT), 4-Pentynoic acid, and Sephadex G10, G15, and G25, dialysis tubing MWCO 2 kDa were purchased from Sigma Aldrich and used as received. NH2-PEG2000-CC was obtained as previously described [29 (link)].
MCF-7 and MDA-MB-231 cell lines were purchased from Sigma Aldrich and cultured in supplemented Dulbecco’s Minimum Essential Medium (DMEM) supplemented with 10% fetal bovine serum (FBS, EuroClone, Milan, Italy), 1% of penicillin/streptomycin (10,000 U mL⁻¹ and 10 mg mL⁻¹ respectively, EuroClone), and 1% of l-glutamine (EuroClone), at 37 °C in 5% CO₂ humidified atmosphere. Cell Titer 96 Aqueous One Solution Cell Proliferation assay (MTS solution) was purchased from Promega (Madison, WI, USA).

Nicosia A., Cavallaro G., Costa S., Utzeri M.A., Cuttitta A., Giammona G, & Mauro N. (2020). Carbon Nanodots for On Demand Chemophotothermal Therapy Combination to Elicit Necroptosis: Overcoming Apoptosis Resistance in Breast Cancer Cell Lines. Cancers, 12(11), 3114.

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Mitochondrial Dysfunction and Oxidative Stress Assays

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Irinotecan hydrochloride, 2-thenoyltrifluoroacetone (TTFA), hydroxyurea (HU), 2′,7′-dichlorofluorescin diacetate (DCFDA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2-deoxy-d-glucose (2-DG), crystal violet, and antimycin A were purchased from Sigma Aldrich (Deisenhofen, Germany). Doxorubicin was purchased from Enzo Life Science (Lörrach, Germany). Chloramphenicol was purchased from Carl Roth GmbH (Karlsruhe, Germany). Rotenone, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP), and oligomycin were purchased from Abcam (Cambridge, UK). 3,3′-Dihexyloxacarbocyanine iodide (DiOC₆(3)) was purchased from Thermo Fisher Scientific (Waltham, MA, USA). DMSO was used as a treatment control.

Marx C., Sonnemann J., Maddocks O.D., Marx-Blümel L., Beyer M., Hoelzer D., Thierbach R., Maletzki C., Linnebacher M., Heinzel T, & Krämer O.H. (2022). Global metabolic alterations in colorectal cancer cells during irinotecan-induced DNA replication stress. Cancer & Metabolism, 10, 10.

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Preparation of Chemotherapeutic Agents

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Irinotecan hydrochloride, folinic acid calcium salt hydrate, and 5-fluorouracil were purchased from Sigma (St. Louis, MO, USA). All drugs were prepared as 1 mM stock solutions by dissolving the compound in nanoPure wa-ter. The 1 mM solution was further diluted in medium to a final concentration of 20.6 μM, 11.3 μM and 68.5 μM for irinotecan, folinic acid and 5-fluorouracil respectively.

LaBonia G.J., Ludwig K.R., Mousseau C.B, & Hummon A.B. (2017). iTRAQ Quantitative Proteomic Profiling and MALDI-MSI of Colon Cancer Spheroids Treated with Combination Chemotherapies in a 3D Printed Fluidic Device. Analytical chemistry, 90(2), 1423-1430.

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Photochemical Hydrogel Encapsulation

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Irgacure
2959 (2-hydroxy-1-[4-(2-hydroxyethoxy)
phenyl]-2-methyl-1-propanone; I-2959) was kindly provided by Ciba
Specialty Chemicals (Basel, Switzerland). PEGDA 700, alginic acid
sodium salt from brown algae (low viscosity), glycerol, paraformaldehyde,
ethanol, bisbenzimide H 33258, Dulbecco’s phosphate buffered
saline (DPBS) with CaCl₂ and MgCl₂, fetal bovine
serum (FBS), calcium chloride hexahydrate, sodium azide, and irinotecan
hydrochloride were purchased from Sigma-Aldrich (St. Louis, MO). Fluorescence
activated cell sorter (FACS) buffer was prepared as 1% PBS, 5% FBS,
and 0.05% 3 M NaN₃. Human U251 malignant glioma (U251 MG)
cells were a kind gift from Dr. Lena Al-Harthi at Rush University
and were cultured in DMEM (Corning, NY) supplemented with 5% FBS.
HUVECs and EBM-2 medium along with supplements and growth factors
for the cells were purchased from Lonza (Walkersville, MD). Alexa
Fluor 647 conjugated mouse antihuman CD309 (VEGFR-2) antibody and
Alexa Fluor 647 mouse IgG1 (κ isotype control; FC) antibody
were purchased from Biolegend (San Diego, CA). MTS cell proliferation
assay was purchased from Promega (Madison, WI). Water used in all
experiments was deionized to 18.2 MΩ·cm (Nanopure II, Barnstead,
Dubuque, IA). All chemicals were purchased at standard grades and
used as received.

Sharma V., Köllmer M., Szymusiak M., Nitsche L.C., Gemeinhart R.A, & Liu Y. (2014). Toroidal-Spiral Particles for Codelivery of Anti-VEGFR-2 Antibody and Irinotecan: A Potential Implant to Hinder Recurrence of Glioblastoma Multiforme. Biomacromolecules, 15(3), 756-762.

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Biochemical Assay Reagents Protocol

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SN-38 was purchased from Abcam; netropsin and irinotecan hydrochloride were purchased from Sigma. Chlorambucil, podofilox and mannitol were from MicroSource Discovery Systems. TopoII inhibitor ICRF-193 [meso-4,4′-(3,2-butanediyl)-bis(2,6-piperazinedione)] was from Sigma.

Peter S., Yu H., Ivanyi-Nagy R, & Dröge P. (2016). Cell-based high-throughput compound screening reveals functional interaction between oncofetal HMGA2 and topoisomerase I. Nucleic Acids Research, 44(22), e162.

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Irinotecan Preparation and Dilution

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Irinotecan hydrochloride was purchased from Sigma (St. Louis, MO, USA). A 200 μM solution of irinotecan was prepared by dissolving the drug in nanoPure water. The 200 μM solution was further diluted in McCoy’s 5A medium to a final concentration of 50 μM for initial irinotecan penetration studies and to a final concentration of 75 μM for irinotecan metabolism studies.

LaBonia G.J., Lockwood S.Y., Heller A.A., Spence D.M, & Hummon A.B. (2016). Drug penetration and metabolism in 3-dimensional cell cultures treated in a 3D printed fluidic device: Assessment of irinotecan via MALDI imaging mass spectrometry. Proteomics, 16(11-12), 1814-1821.

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Comprehensive Protocol for B16F10 Melanoma Cell Culture and Analysis

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B16F10 mouse melanoma cancer cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Sodium alginate (Protanal LF20/40) of high molecular weight (~250 kDa) was donated by FMC BioPolymers (Philadelphia, PA, USA). Trypan Blue, Dulbecco’s modified eagle’s medium (DMEM), bovine serum albumin (BSA), fetal bovine serum (FBS), penicillin-streptomycin, and trypsin-EDTA solutions, morpholineethanesulfonic acid (MES) hydrate, Dulbecco’s phosphate buffered saline (DPBS), Sigmacote, activated charcoal, calcium sulfate dihydrate, irinotecan hydrochloride, 5-fluorouracil, mitoxantrone hydrochloride, fluorescein isothiocyanate (FITC) dextran (3–6 kDa and 70 kDa), fluorescein isothiocyanate diethylaminoethyl (FITC-DEAE) dextran (3–6 kDa and 70 kDa) and rhodamine dextran (10 kDa) were all purchased from Sigma-Aldrich (St. Louis, MO, USA). Vascular endothelial growth factor (VEGF) and Platelet-derived growth factor (PDGF) proteins and VEGF- and PDGF-DuoSet ELISA kits were purchased from R&D Systems, Inc (Minneapolis, MN, USA). 16-well xCELLigence e-plates were purchased from ACEA Biosciences, Inc. (San Diego, CA, USA).

Emi T., Michaud K., Orton E., Santilli G., Linh C., O’Connell M., Issa F, & Kennedy S. (2019). Ultrasonic Generation of Pulsatile and Sequential Therapeutic Delivery Profiles from Calcium-Crosslinked Alginate Hydrogels. Molecules, 24(6), 1048.

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Cytotoxic Effects of β-LG and Irinotecan

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β-LG and irinotecan hydrochloride were prepared from Sigma-Aldrich Co. (St. Louis, Mo, USA). NaOH, KH2PO4, and NaCl were of analytical grade from Merck (Darmstadt, Germany). 3-(4,5- dimethyl- thiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich Co. (St. Louis, Mo, USA). The HT29 (human colon cancer cell line) and AGS (gastric adenocarcinoma cell line) were purchased from the National Cell Bank, Pasteur Institute of Iran, Tehran, I.R. Iran. All other chemicals used in this study were of analytical grade.

Bijari N., Ghobadi S, & Derakhshandeh K. (2019). β-lactoglobulin-irinotecan inclusion complex as a new targeted nanocarrier for colorectal cancer cells. Research in Pharmaceutical Sciences, 14(3), 216-227.

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Chiral Propanediol and Irinotecan Characterization

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Enantiomers (S)-(+)-1,2-Propanediol and (R)-(−)-1,2-Propanediol were used as received (Sigma-Aldrich, products 540242 and 540250, 96%). The Propanediol enantiomers were prepared with a flow-cell with thickness of ∼70 μm. The anticancer drug (Sigma-Aldrich, Irinotecan hydrochloride) was dissolved in water and formed a solution with concentration of 1 mg ml⁻¹. The anticancer drug experiment was also performed with the flow-cell. The flow-cell was prepared and replaced every time for each measurement. The protein was dissolved in 10 mM Tris/HCl buffer solution with a controlled pH value at 7, forming a concentration of 1 mg ml⁻¹. The prepared protein solution was spin-coated onto the clean metamaterial sample at a spin speed of 2,000 r.p.m., forming a monolayer of ∼10 nm, which is confirmed with the ellipsometry measurements (J.J. Woollam M-2000 DI).

Zhao Y., Askarpour A.N., Sun L., Shi J., Li X, & Alù A. (2017). Chirality detection of enantiomers using twisted optical metamaterials. Nature Communications, 8, 14180.

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Irinotecan, MG132, and X-ray Treatments

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Where indicated, HCT116 cells were treated with irinotecan hydrochloride (Sigma) at concentrations stated. U2OS cells were treated with 20 μM MG132 for 4 hours (Calbiochem) or Leptomycin B (LMB) (Calbiochem) at 20 nM for 3 hours where indicated. For irradiation, cells were exposed to X-rays at 1 Gy/min (250 kV constant potential, Pantak X-ray machine).

Sabir S.R., Sahota N.K., Jones G.D, & Fry A.M. (2015). Loss of Nek11 Prevents G2/M Arrest and Promotes Cell Death in HCT116 Colorectal Cancer Cells Exposed to Therapeutic DNA Damaging Agents. PLoS ONE, 10(10), e0140975.

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