Paclitaxel

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

Paclitaxel is a cytotoxic compound that is used in the production of various pharmaceutical and laboratory products. It is a naturally occurring substance extracted from the Pacific yew tree. Paclitaxel inhibits cell division by stabilizing microtubules, which are essential for cell division and growth.

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Lab products found in correlation

50 protocols using paclitaxel

Cell Culture of Gastric Cancer Lines

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The human normal gastric epithelial cell line GES-1, GC cell lines BGC823 and SGC7901 and HEK293T cells were obtained from China Academy of Chinese Medical Sciences. The human GC SGC7901/PA cell line (resistance to paclitaxel) was obtained from the Fourth Military Medical University (Xi’an, Shanxi, China). These cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), and cultured with 5% CO₂ at 37°C. In order to maintain the sensitivity phenotype, SGC7901/PA cells was cultured with 20 μg/l paclitaxel (Gibco, Carlsbad, CA, USA), and cultured in paclitaxel-free medium for 14 days prior to the experiment.

Zhao B., Zhang J., Chen X., Xu H, & Huang B. (2018). Mir-26b inhibits growth and resistance to paclitaxel chemotherapy by silencing the CDC6 gene in gastric cancer. Archives of Medical Science : AMS, 15(2), 498-503.

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Chemotherapeutic-Induced Peripheral Neuropathy Treatment

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Chemotherapeutics including paclitaxel, oxaliplatin and vincristine (Fisher Scientific, Pittsburgh, PA) were administered intraperitoneally (i.p.) per the following dose regimens: paclitaxel 2 mg/kg i.p. four times on alternate days (1, 3, 5, 7), vincristine 0.1 mg/kg i.p. over 12 days, with two 5-day cycles and a 2-day break in between and, oxaliplatin once at 6 mg/kg i.p. The sEHI EC5026 and EC5029 treatments tested on the established CIPN models were formulated as true solutions in PEG400 (Sigma Aldrich, St. Louis MO) which was tested as the vehicle, and all compounds were administered by oral gavage. Morphine sulfate was formulated in saline (Fisher Scientific, Pittsburgh, PA) and administered by subcutaneous injection. Pregabalin (Tocris, Minneapolis, MN) was formulated in water and administered by oral gavage.

Takeshita A.A., Hammock B.D, & Wagner K.M. (2023). Soluble epoxide hydrolase inhibition alleviates chemotherapy induced neuropathic pain. Frontiers in Pain Research, 3, 1100524.

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Synergistic Inhibition of TNBC Proliferation

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To determine whether riluzole and paclitaxel synergistically inhibit proliferation of TNBC cells, MTT assays were performed as previously described [13 (link)]. Briefly, cells were treated with riluzole (Sigma-Aldrich, 1–50 μM) and paclitaxel (Invitrogen, 0.5–25 nM), at constant ratio of 1:2000 (riluzole:paclitaxel). Cell viability was determined on day 3 by MTT assay [13 (link)]. In some experiments, cell numbers were determined in parallel with the MTT assay by counting manually on a hemocytometer. To assess the interaction between riluzole and paclitaxel, Compusyn 1.0 software was used to generate isoboles. Using this method of isoboles [21 (link)], the dose-effect data of individual drugs measured above was used to determine the expected combination and then statistically compared to the actual combination effect measured to determine synergism, additivity, or anti-additive interactions. The combination index (CI) at different F_a levels was also determined using this software based on the following equation:

CI = (D) 1 /(D x)1 + (D) 2 /(D x) 2 .

Speyer C.L., Bukhsh M.A., Jafry W.S., Sexton R.E., Bandyopadhyay S, & Gorski D.H. (2017). Riluzole synergizes with paclitaxel to inhibit cell growth and induce apoptosis in triple-negative breast cancer. Breast cancer research and treatment, 166(2), 407-419.

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Establishing Paclitaxel-Resistant Breast Cancer Cells

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Human breast cancer cell lines MDA‐MB‐231 and MCF‐7 were acquired from the American Type Culture Collection (Beijing Zhongyuan Ltd) and cultured in media as recommended by the provider. Paclitaxel‐resistant MDA‐MB‐231 cells were cultured in Dulbecco's modified Eagle's medium (Gibco, ThermoFisher Scientific) supplemented with 10% foetal bovine serum (Hyclone, ThermoFisher Scientific). The Paclitaxel‐resistant cell line is derived from parental MDA‐MB‐231 cells. MDA‐MB‐231 cells were subjected to increasing concentrations of Paclitaxel (T1912, Sigma) until the cells acquire resistance to 0.2 mol/L of Paclitaxel. The Paclitaxel resistance cell was then maintained in 0.2 mol/L of Paclitaxel. For double thymidine block, cells were arrested with 2 mmol/L thymidine (Sigma) medium for 19 hours. After cells were washed three times with phosphate‐buffered saline (PBS) and released into fresh medium for 9 hours, they were then treated with thymidine for an additional 16 hours. To synchronize cells in G2/M phase, cells were treated with 40 ng/mL nocodazole (Sigma) for 16 hours.

Yang N., Wang C., Wang J., Wang Z., Huang D., Yan M., Kamran M., Liu Q, & Xu B. (2019). Aurora kinase A stabilizes FOXM1 to enhance paclitaxel resistance in triple‐negative breast cancer. Journal of Cellular and Molecular Medicine, 23(9), 6442-6453.

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Evaluating Anticancer Drug Efficacy

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Cells were seeded in 96 well-plates, with or without paclitaxel (#Y0000698, Sigma-Aldrich), doxorubicin hydrochloride (#D2975000, Sigma-Aldrich), tamoxifen (#HT904, Sigma-Aldrich), or fulvestrant (#S1191, Selleckchem). Medium was replaced every two days. After 72 h for paclitaxel and doxorubicin, and seven days for tamoxifen and fulvestrant, to assess the proliferative effect of estrogen, cells were cultured in 24-well plates, in phenol red-free DMEM:F12 medium (# 11039021, Gibco) supplemented with 5% (v/v) charcoal stripped FBS (csFBS, #12676029, Gibco), for five days, with or without 10 nM β-estradiol (#E2758, Sigma-Aldrich). Viability was assessed by Alamar blue assay (#DAL1100, Invitrogen).

Gomes I., de Almeida B.P., Dâmaso S., Mansinho A., Correia I., Henriques S., Cruz-Duarte R., Vilhais G., Félix P., Alves P., Corredeira P., Barbosa-Morais N.L., Costa L, & Casimiro S. (2020). Expression of receptor activator of NFkB (RANK) drives stemness and resistance to therapy in ER+HER2- breast cancer. Oncotarget, 11(19), 1714-1728.

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Chemical Library Acquisition and Verification

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The chemical library was donated by the EPA ToxCast Program. For verification and replication experiments, the following chemicals were purchased from Sigma-Aldrich: DL-α tocopherol acetate (T3376), DL-sulforophane (S4441), vincristine sulfate salt (V8879), oxyfluorfen (35031), rotenone (45656), fenamidone (33965), pyraclostrobin (33696), trifloxystrobin (46447), myxothiazol (T5580), pyridaben (46047), azoxystrobin (31697), fluoxastrobin (33797), fenpyroximate (31684) and kresoxim-methyl (37899). Famoxadone was purchased from Chem Service, Inc (N-11943). Topotecan hydrochloride was purchased from Tocris (4562). Paclitaxel was purchased from Fisher Scientific (AC32842). All chemical stocks were prepared in DMSO unless otherwise noted. Vehicle samples were prepared with an equivalent DMSO concentration of ≤0.5% in feeding medium.

Pearson B.L., Simon J.M., McCoy E.S., Salazar G., Fragola G, & Zylka M.J. (2016). Identification of chemicals that mimic transcriptional changes associated with autism, brain aging and neurodegeneration. Nature Communications, 7, 11173.

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Analytical Techniques for Paclitaxel and Cisplatin

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We purchased cis-diammineplatinum (II) dichloride (cisplatin) from Sigma-Aldrich (St. Louis, MO, USA), and paclitaxel was donated from Samyang Biopharmaceuticals Corp. (Seongnam, Korea). For analyzing serum and tissue concentrations of paclitaxel and cisplatin, we purchased acetonitrile and methanol from Fisher Scientific (Waltham, MA, USA) and formic acid, acetic acid, ammonium diethyldithiocarbamate, and ammonium acetate from Sigma-Aldrich.

Park S.J., Lee E.J., Seol A., Park S., Ham J., Yim G.W., Shim S.H., Lim W., Chang S.J., Song G., Park J.W, & Kim H.S. (2022). Rotational intraperitoneal pressurized aerosol chemotherapy with paclitaxel and cisplatin: pharmacokinetics, tissue concentrations, and toxicities in a pig model. Journal of Gynecologic Oncology, 33(5), e56.

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Synthesis and Characterization of MDOR Antagonist D24M

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The MDOR antagonist D24M was synthesized, assessed for >95% purity by HPLC, and structure confirmed by HRMS as reported in our previous work [34 ]. Morphine sulfate pentahydrate was obtained from the NIDA Drug Supply Program. Oxymorphone (#0790) and buprenorphine (#3210) were obtained from Mallinckrodt (St. Louis, MO). CTAP (#15–601), Naltrindole (#50–178-9293), Naloxonazine (#05–911-0), KN-93 (#52–151), Src-I1 (#36–421-0R), and paclitaxel (#AAJ62734MC) were all obtained from Fisher Scientific (Waltham, MA). D24M, KN-93, and Src-I1 were prepared as 10 mM stocks in DMSO, and stored at −20°C. CTAP, Naloxonazine, and Naltrindole were prepared as 10 mM stocks in USP sterile water and stored at −20°C. Oxymorphone, buprenorphine, and morphine were prepared fresh prior to every experiment in USP sterile saline. paclitaxel was prepared fresh for every experiment and dissolved in a vehicle of 16.7% cremophor, 16.7% ethanol, and 66.6% USP sterile saline. Matched vehicle control injections were included in every experiment: 2% DMSO, 10% Tween80, 88% USP water for D24M, KN-93, and Src-I1; USP water for CTAP, Naloxonazine, and Naltrindole; USP saline for morphine, buprenorphine, and oxymorphone. Drug powders were stored as recommended by the manufacturer, and D24M powder was stored at −80°C under desiccation.

Keresztes A., Olson K., Nguyen P., Lopez-Pier M.A., Hecksel R., Barker N.K., Liu Z., Hruby V., Konhilas J., Langlais P.R, & Streicher J.M. (2022). Antagonism of the Mu-Delta Opioid Receptor Heterodimer Enhances Opioid Anti-Nociception by Activating Src and CaMKII Signaling. Pain, 163(1), 146-158.

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Mass Spectrometry Sample Preparation

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2,5-Dihydroxybenzoic acid (DHB)
and trifluoroacetic acid (TFA) were purchased from Sigma-Aldrich (Zwijndrecht,
The Netherlands). Ultrapure water (LC-MS grade) and methanol (MeOH;
HPLC grade) were purchased from Fisher Scientific (Loughborough, Leicestershire,
U.K.). Ketoconazole, terfenadine, haloperidol, doxorubicin, paclitaxel,
kynurenine, triamcinolone acetonide (TAA), caffeine, fluoxetine, and
ibuprofen standards were originally purchased from Fisher Scientific
(Loughborough, Leicestershire, U.K.). Dulbecco’s modified Eagle’s
medium (DMEM) and penicillin/streptomycin (P/S) were purchased from
Thermo Fisher Scientific (Waltham, MA, U.S.A.). Fetal bovine serum
(FBS) was purchased from HyClone (Eindhoven, The Netherlands) and
ammonium acetate was purchased from AMRESCO (Solon, OH, U.S.A.). Hematoxylin
solution modified according to Gill and Entellan new was purchased
from Merck KGaA (Darmstadt, Germany). Safranin-O and Fast Green were
purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). Eosine-Y,
alcoholic, was purchased from Avantor Performance Materials B.V. (Deventer,
The Netherlands), and coverslips were purchased from Thermo Scientific
(Waltham, Massachusetts, U.S.A.).

Barré F.P., Paine M.R., Flinders B., Trevitt A.J., Kelly P.D., Ait-Belkacem R., Garcia J.P., Creemers L.B., Stauber J., Vreeken R.J., Cillero-Pastor B., Ellis S.R, & Heeren R.M. (2019). Enhanced Sensitivity Using MALDI Imaging Coupled with Laser Postionization (MALDI-2) for Pharmaceutical Research. Analytical Chemistry, 91(16), 10840-10848.

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Diverse Pharmacological Agents in Research

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Drugs, ligands and inhibitors used in this study include etoposide (Sigma Aldrich, St. Louis, MO, USA), SN-38 (Cayman Chemical, Ann Arbor, MI, USA), gemcitabine (Sigma Aldrich), carbonyl cyanide m-chlorophenylhydrazone (CCCP, Sigma Aldrich), phorbol 12-myristate 13-acetate (PMA, Sigma Aldrich), bryostatin 1 (Tocris, Bristol, UK), Ro 31-8220 (Tocris), TPCA-1 (Sigma Aldrich), recombinant human IL-4 (PharMingen, Franklin Lakes, NJ, USA) and paclitaxel (LC Labs, Woburn, MA, USA). All drugs and inhibitors were dissolved in DMSO (Fisher, Waltham, MA, USA) except recombinant human IL-4 which was dissolved in water and paclitaxel which was dissolved 50:50 Kolliphor/ethanol.

Fermaintt C.S., Peramuna T., Cai S., Takahashi-Ruiz L., Essif J.N., Grant C.V., O’Keefe B.R., Mooberry S.L., Cichewicz R.H, & Risinger A.L. (2021). Yuanhuacine Is a Potent and Selective Inhibitor of the Basal-Like 2 Subtype of Triple Negative Breast Cancer with Immunogenic Potential. Cancers, 13(11), 2834.

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