Journal: Cells

Article Title: A Drug Repurposing Screen Identifies Fludarabine Phosphate as a Potential Therapeutic Agent for N-MYC Overexpressing Neuroendocrine Prostate Cancers

doi: 10.3390/cells11142246

Figure Lengend Snippet: High-throughput drug library screen identified potential lead compounds that selectively inhibits proliferation MYCN overexpressing cells. ( A ) Scatter plot graphs show that fludarabine/fludarabine phosphate is a lead compound eliminating MYCN overexpressing cells. FDA-approved drug library compounds were applied at three different concentrations: 100 nM top panel, 500 nM middle panel, and 1 mM bottom panel to LNCap ( x -axis) and LNCap-N-Myc ( y -axis) cells and the cells were monitored for five days using high content imaging. Fludarabine/fludarabine phosphate red dots, additional hits blue dots, and non-hit grey dots. ( B ) Heatmap plot shows fludarabine and fludarabine phosphate exhibit preferential inhibition of LNCap-N-Myc cells at 100 nM, 500 nM, and 1 mM concentrations over five days (D1, D2, D3, D4, and D5). Yellow squares represent the days, where significant differences were identified. Hits in green text at 100 nM, hits in blue text at 500 nM, and hits in orange text at 1 mM ( p < 0.05).

Article Snippet: Fludarabine phosphate (#S1229, Selleckchem, Houston, TX, USA).

Techniques: High Throughput Screening Assay, Drug discovery, Imaging, Inhibition

Journal: Cells

Article Title: A Drug Repurposing Screen Identifies Fludarabine Phosphate as a Potential Therapeutic Agent for N-MYC Overexpressing Neuroendocrine Prostate Cancers

doi: 10.3390/cells11142246

Figure Lengend Snippet: Fludarabine phosphate affects MYC protein levels and MYC transcriptional targets in NEPC cells. ( A ) Confluency measurements of LASCPC-01 cells following treatment of fludarabine phosphate 112 h after seeding (*** p < 0.001). ( B ) Fludarabine phosphate treatment induces apoptosis in LASCPC-01 cells at 25 μM. Western blot shows and PARP. b tubulin is used as a loading control. ( C ) Fludarabine phosphate causes DNA damage. Western blots show an increase in H2A.X in a dose-and-time-dependent manner in LASCPC-01 cells. Cells were treated with 25 μM fludarabine phosphate in the time course experiment. ( D ) Western blot shows a concentration-dependent decrease in N-MYC in fludarabine phosphate-treated LASCPC-01 cells. Cells were treated with DMSO or indicated concentrations of fludarabine phosphate for 48 h. Whole-cell extracts were subjected to Western blot analysis for N-MYC, NSE, and SYP. ImageLab software was used to quantify the bands with corresponding GAPDH and represented in the above blots. ( E ) Western blot shows time course analysis of N-Myc, NSE, and SYP levels at 48 h of 25 μM fludarabine phosphate treatment compared to the DMSO control at 48 h. ( F ) The e pression level of MYCN by RT-PCR in fludarabine phosphate-treated LASCPC-01 cells. Cells were treated with DMSO or indicated concentrations of fludarabine phosphate for 48 h. n.s not significant. ( G ) Volcano plot showing fold change in gene expression between fludarabine phosphate treated with 25 μM and untreated LASCPC-01 cells and their corresponding p -values at 48 h. ( H ) RNAseq expression analysis shows that 64 MYC-target genes were downregulated upon treatment with fludarabine phosphate in LASCPC-01 cells at 48 h compared to the DMSO-treated control.

Article Snippet: Fludarabine phosphate (#S1229, Selleckchem, Houston, TX, USA).

Techniques: Western Blot, Control, Concentration Assay, Software, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Expressing

Journal: Cells

Article Title: A Drug Repurposing Screen Identifies Fludarabine Phosphate as a Potential Therapeutic Agent for N-MYC Overexpressing Neuroendocrine Prostate Cancers

doi: 10.3390/cells11142246

Figure Lengend Snippet: Fludarabine phosphate destabilizes N-MYC protein by inhibiting AKT signaling. ( A ) Fludarabine phosphate inhibits AKT activation. LASCPC-01 cells were pretreated with the ROS scavenger, NAC for 3 h, then treated with fludarabine phosphate at 25 μM for 48 h. Whole-cell extracts were subjected to Western blot analysis for phospho Akt and Akt. Cell signaling antibodies AKT #9272, phospho-AKT Ser473 #9271 against AKT and phosphoAkt used in this study detect AKT1,2 and 3. For phospho-Akt, short exposure and long exposure are presented. ( B ) Fludarabine phosphate triggers NMYC downregulation. LASCPC-01 cells were treated as in ( A ) and whole-cell extracts were subjected to Western blot analysis for N-Myc and representation from one of the three biological replicates is presented. ( C ) Fludarabine phosphate effect on pMEK. LASCPC-01 cells were treated as in ( A ) and whole-cell extracts were subjected to Western blot analysis for pMEK and representation from one of the three biological replicates is presented. ( D ) Fludarabine phosphate targets Akt proteins for proteasome-mediated degradation. LASCPC-01 cells were pre-treated for three hours with bortezomib (BZM) at 5 nM or matching volume of solvent, then treated with fludarabine phosphate at 25 μM for 48 h. Whole-cell extracts were analyzed for total AKT levels by Western blotting. ( E ) Quantitation of Akt level (the upper band *) of the WB in figure E, from two biological replicates. ImageLab software was used to quantify the bands. Akt p values 0.0153 and 0.0198 for FP-treated and BZM/FP, respectively. ( F ) Schematic illustration of the effect of fludarabine phosphate on N-MYC overexpressing cells.

Article Snippet: Fludarabine phosphate (#S1229, Selleckchem, Houston, TX, USA).

Techniques: Activation Assay, Western Blot, Solvent, Quantitation Assay, Software

Journal: Cells

Article Title: A Drug Repurposing Screen Identifies Fludarabine Phosphate as a Potential Therapeutic Agent for N-MYC Overexpressing Neuroendocrine Prostate Cancers

doi: 10.3390/cells11142246

Figure Lengend Snippet: Fludarabine phosphate reduces tumor growth in NEPC xenograft models. ( A ) Validation of fludarabine phosphate treatment in 22Rv1, 22Rv1-N-Myc and LASCPC-01 xenograft models. Each cell line was implanted at 1 × 10 6 cells subcutaneously by injection in PBS: Matrigel mixture at the 1: 1 ratio of in a total volume of 100 μL and monitored for tumor development. Fludarabine phosphate or DMSO was administered through i.p. injections at 120mg/kg starting day 6–7 post cell lines engraftment. Total of 10 i.p. injections were given per mouse (one i.p. injection per mouse per day was given for five consecutive days followed by two days rest and then, treatment continued for another five i.p. injections over five days). Tumors were measured every 3–4 days using digital caliper, and tumor volume was calculated using the formula A/2*B2 where A and B represent the long and short diameter of the tumor, respectively. Two biological experiments were performed n = 2 with 4 to 5 mice per group and the graph represents one biological replicate. (* p < 0.05; ** p < 0.01). ( B ) Graphs show body weights of fludarabine phosphate treated and DMSO-control mice. Mice were weighed every 3–4 days to assess potential side effects of the fludarabine phosphate administration. ( C ) Fludarabine phosphate reduces N-Myc protein levels in tumor tissue. Upon experiment termination, tumor tissues were collected from the LASCPC-01 xenograft model, tumor tissue homogenates were prepared, and Western blot analysis were performed for the indicated proteins in individual tumors. Western blot analysis was performed from individual tumor samples. ( D ) Quantitation of Western blots (* p < 0.05; ** p < 0.01) ( C ), Image Lab software was used to quantify the bands.

Article Snippet: Fludarabine phosphate (#S1229, Selleckchem, Houston, TX, USA).

Techniques: Biomarker Discovery, Injection, Control, Western Blot, Quantitation Assay, Software

Journal: Gene therapy

Article Title: Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics

doi: 10.1038/gt.2010.103

Figure Lengend Snippet: Systemic treatment with virus and fludarabine reduces tumor burden. (A) Time course of tumor growth in four groups of 8–9 mice treated as indicated in the text. Curves are shown until the time when two mice in a group had to be sacrificed. (B) Tumor volumes of individual mice 24 days after the beginning of the treatment. The average tumor volume is indicated by an horizontal line; error bars indicate the standard deviation.

Article Snippet: Fludarabine (phosphate for injection) (SICOR Pharmaceuticals, Irvine, CA) was reconstituted with sterile H 2 O as per manufacturer.

Techniques: Virus, Standard Deviation

Journal: Gene therapy

Article Title: Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics

doi: 10.1038/gt.2010.103

Figure Lengend Snippet: Time course of tumor growth in mice treated with fludarabine at different intervals after virus delivery (A) and Kaplan Meier survival curve of the same mice (B). Groups of 9–11 mice were treated with fludarabine immediately after virus administration (F1), or after a one week interval (F2), or a two weeks interval (F3). In (A) curves are shown until the time when two mice in a group had to be sacrificed. In (B) the defined end point was a 1.5 ml tumor volume.

Article Snippet: Fludarabine (phosphate for injection) (SICOR Pharmaceuticals, Irvine, CA) was reconstituted with sterile H 2 O as per manufacturer.

Techniques: Virus

Journal: Gene therapy

Article Title: Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics

doi: 10.1038/gt.2010.103

Figure Lengend Snippet: Triple regimen chemovirotherapy. (A) Time course of tumor growth in mice treated with different combinations of CPA, virus and fludarabine. (B) Individual tumor volumes at day 20 after virus administration. Groups of 10–12 mice were treated. Timing of administration of the three treatments is as detailed in the main text. (C) Percentile of Granta cells in white blood cells (wbc) on day 25 after start of treatment. The six treatment groups are shown using the same symbols in each panel.

Article Snippet: Fludarabine (phosphate for injection) (SICOR Pharmaceuticals, Irvine, CA) was reconstituted with sterile H 2 O as per manufacturer.

Techniques: Virus

Journal: Gene therapy

Article Title: Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics

doi: 10.1038/gt.2010.103

Figure Lengend Snippet: Repeated sequential chemovirotherapy leads to complete regression of implanted tumors and long-term survival. (A) Time course of tumor growth in mice treated with different regimens of three therapeutics. Groups of about 12 mice were treated, but fatalities due to repeated injections caused some losses in each group, as detailed in . The schedule of each treatment is indicated below the time axis. C: CPA. MV: measles virus. F: fludarabine. (B) Kaplan-Meier survival curves. Symbols are as in panel A. The defined end point was 1,700 μl tumor volume. After day 52, no mice had to be sacrificed because of excessive tumor volume.

Article Snippet: Fludarabine (phosphate for injection) (SICOR Pharmaceuticals, Irvine, CA) was reconstituted with sterile H 2 O as per manufacturer.

Techniques: Virus

Journal: PLoS ONE

Article Title: Efficiency of four solutions in removing 23 conventional antineoplastic drugs from contaminated surfaces

doi: 10.1371/journal.pone.0235131

Figure Lengend Snippet: Efficiency of the decontamination solutions on the 23 tested antineoplastic drugs on stainless steel surfaces after standard single motion. Solution 1 (S1): 70% isopropanol; Solution 2 (S2): ethanol (91.6 mg/g) hydrogen peroxide (50.0 mg/g); Solution 3 (S3): 10 −2 M sodium dodecyl sulfate:isopropanol 80:20; Solution 4 (S4): 0.5% sodium hypochlorite aqueous solution. 5FU: 5-fluorouracil; Cyta: cytarabine; Fluda: fludarabine; Ganci: ganciclovir; Gem: gemcitabine; Mtx: methotrexate; Peme: pemetrexed; Ralti: raltitrexed; Busu: busulfan; Cyc: cyclophosphamide; Ifos: ifosphamide; Dacar: dacarbazine; Dauno: daunorubicin; Doxo: doxorubicin; Ida: idarubicin; Epi: epirubicin; EtopoP: etoposide phosphate; Eto: etoposide; Dtx: docetaxel; Pcx: paclitaxel; Irino: irinotecan; Topo: topotecan; Vin: vincristine.

Article Snippet: The ICADs used for experiments were: 5-fluorouracil (batch# PE4SH-RM), methotrexate (batch# G07UG-01) and dacarbazine (batch# Z3J8O-SF), purchased from Tokyo Chemical Industry (Zwijndrecht, Belgium); gemcitabine (batch# A0375170), purchased from Acros Organic (Geel, Belgium); busulfan (batch# BCBN8120V), purchased from Sigma-Aldrich (Buchs, Switzerland); ganciclovir, purchased from Roche Pharma (Cymevene ® , batch# B4091B08, Reinach, Switzerland); cytarabine (batch# 3-YFD-59-1), epirubicin (batch# 11-CGS-118-1) and topotecan (batch# 8-MSW-162-1) purchased from Toronto Research Chemicals (North York, ON, Canada); raltitrexed (batch# MG10453-29102016), pemetrexed (batch# AGN2017-685), docetaxel (batch# CS13527-18112016), paclitaxel (batch# CS22539-20112016), vincristine sulfate (batch# MC-10452-0212016), doxorubicin hydrochloride (batch# MC10454-0612016), daunorubicin (batch# MC10456-1712016), idarubicin (batch# 5-CGS-96-1), etoposide (batch# MC10457-20102016), etoposide phosphate (batch# MC305547-21082017), irinotecan hydrochloride (batch# MC20783-21082017) and fludarabine phosphate (batch# MC444607-21082017) brought from Pharmaserv (Sansstad, Switzerland).

Techniques:

Journal: PLoS ONE

Article Title: Efficiency of four solutions in removing 23 conventional antineoplastic drugs from contaminated surfaces

doi: 10.1371/journal.pone.0235131

Figure Lengend Snippet: Values represented are median Effq for each drug (n = 6). 5FU: 5-fluorouracil; Cyta: cytarabine; Fluda: fludarabine; Ganci: ganciclovir; Gem: gemcitabine; Mtx: methotrexate; Peme: pemetrexed; Ralti: raltitrexed; Busu: busulfan; Cyc: cyclophosphamide; Ifos: ifosphamide; Dacar: dacarbazine; Dauno: daunorubicin; Doxo: doxorubicin; Ida: idarubicin; Epi: epirubicin; EtopoP: etoposide phosphate; Eto: etoposide; Dtx: docetaxel; Pcx: paclitaxel; Irino: irinotecan; Topo: topotecan; Vin: vincristine. Blue line/circles: 70% isopropanol; red line/triangles: admixture of ethanol-hydrogen peroxide (91.6–50.0 mg/g); green line/diamonds: admixture of 10 −2 M sodium dodecyl sulfate/isopropanol (80/20) and yellow line/squares: sodium hypochlorite. * Significant difference for solution 4 over other solutions; ** Significant difference between solutions 4 and 2; $ significant difference between solutions 4 and 1 and between $ $ solutions 4 and 3. + significant difference for both solutions 3 and 4 compared to others.

Article Snippet: The ICADs used for experiments were: 5-fluorouracil (batch# PE4SH-RM), methotrexate (batch# G07UG-01) and dacarbazine (batch# Z3J8O-SF), purchased from Tokyo Chemical Industry (Zwijndrecht, Belgium); gemcitabine (batch# A0375170), purchased from Acros Organic (Geel, Belgium); busulfan (batch# BCBN8120V), purchased from Sigma-Aldrich (Buchs, Switzerland); ganciclovir, purchased from Roche Pharma (Cymevene ® , batch# B4091B08, Reinach, Switzerland); cytarabine (batch# 3-YFD-59-1), epirubicin (batch# 11-CGS-118-1) and topotecan (batch# 8-MSW-162-1) purchased from Toronto Research Chemicals (North York, ON, Canada); raltitrexed (batch# MG10453-29102016), pemetrexed (batch# AGN2017-685), docetaxel (batch# CS13527-18112016), paclitaxel (batch# CS22539-20112016), vincristine sulfate (batch# MC-10452-0212016), doxorubicin hydrochloride (batch# MC10454-0612016), daunorubicin (batch# MC10456-1712016), idarubicin (batch# 5-CGS-96-1), etoposide (batch# MC10457-20102016), etoposide phosphate (batch# MC305547-21082017), irinotecan hydrochloride (batch# MC20783-21082017) and fludarabine phosphate (batch# MC444607-21082017) brought from Pharmaserv (Sansstad, Switzerland).

Techniques:

Journal: PLoS ONE

Article Title: Efficiency of four solutions in removing 23 conventional antineoplastic drugs from contaminated surfaces

doi: 10.1371/journal.pone.0235131

Figure Lengend Snippet: Efficiency of the decontamination solutions on the 23 tested antineoplastic drugs on stainless steel surfaces after vigorous decontamination. Solution 1 (S1): 70% isopropanol; Solution 2 (S2): ethanol (91.6 mg/g) hydrogen peroxide (50.0 mg/g); Solution 3 (S3): 10 −2 M sodium dodecyl sulfate:isopropanol 80:20; Solution 4 (S4): 0.5% sodium hypochlorite aqueous solution. 5FU: 5-fluorouracil; Cyta: cytarabine; Fluda: fludarabine; Ganci: ganciclovir; Gem: gemcitabine; Mtx: methotrexate; Peme: pemetrexed; Ralti: raltitrexed; Busu: busulfan; Cyc: cyclophosphamide; Ifos: ifosphamide; Dacar: dacarbazine; Dauno: daunorubicin; Doxo: doxorubicin; Ida: idarubicin; Epi: epirubicin; EtopoP: etoposide phosphate; Eto: etoposide; Dtx: docetaxel; Pcx: paclitaxel; Irino: irinotecan; Topo: topotecan; Vin: vincristine.

Article Snippet: The ICADs used for experiments were: 5-fluorouracil (batch# PE4SH-RM), methotrexate (batch# G07UG-01) and dacarbazine (batch# Z3J8O-SF), purchased from Tokyo Chemical Industry (Zwijndrecht, Belgium); gemcitabine (batch# A0375170), purchased from Acros Organic (Geel, Belgium); busulfan (batch# BCBN8120V), purchased from Sigma-Aldrich (Buchs, Switzerland); ganciclovir, purchased from Roche Pharma (Cymevene ® , batch# B4091B08, Reinach, Switzerland); cytarabine (batch# 3-YFD-59-1), epirubicin (batch# 11-CGS-118-1) and topotecan (batch# 8-MSW-162-1) purchased from Toronto Research Chemicals (North York, ON, Canada); raltitrexed (batch# MG10453-29102016), pemetrexed (batch# AGN2017-685), docetaxel (batch# CS13527-18112016), paclitaxel (batch# CS22539-20112016), vincristine sulfate (batch# MC-10452-0212016), doxorubicin hydrochloride (batch# MC10454-0612016), daunorubicin (batch# MC10456-1712016), idarubicin (batch# 5-CGS-96-1), etoposide (batch# MC10457-20102016), etoposide phosphate (batch# MC305547-21082017), irinotecan hydrochloride (batch# MC20783-21082017) and fludarabine phosphate (batch# MC444607-21082017) brought from Pharmaserv (Sansstad, Switzerland).

Techniques:

Journal: PLoS ONE

Article Title: Efficiency of four solutions in removing 23 conventional antineoplastic drugs from contaminated surfaces

doi: 10.1371/journal.pone.0235131

Figure Lengend Snippet: Values represented are median Effq for each drug (n = 6). 5FU: 5-fluorouracil; Cyta: cytarabine; Fluda: fludarabine; Ganci: ganciclovir; Gem: gemcitabine; Mtx: methotrexate; Peme: pemetrexed; Ralti: raltitrexed; Busu: busulfan; Cyc: cyclophosphamide; Ifos: ifosphamide; Dacar: dacarbazine; Dauno: daunorubicin; Doxo: doxorubicin; Ida: idarubicin; Epi: epirubicin; EtopoP: etoposide phosphate; Eto: etoposide; Dtx: docetaxel; Pcx: paclitaxel; Irino: irinotecan; Topo: topotecan; Vin: vincristine. Blue line/circles: 70% isopropanol; red line/triangles: admixture of ethanol-hydrogen peroxide (91.6–50.0 mg/g); green line/diamonds: admixture of 10 −2 M sodium dodecyl sulfate/isopropanol (80/20) and yellow line/squares: sodium hypochlorite. + significant difference for both solutions 3 and 4 compared to others.; @ significant differences between the 4 solutions; * significant difference of solution 4 over other solutions.

Article Snippet: The ICADs used for experiments were: 5-fluorouracil (batch# PE4SH-RM), methotrexate (batch# G07UG-01) and dacarbazine (batch# Z3J8O-SF), purchased from Tokyo Chemical Industry (Zwijndrecht, Belgium); gemcitabine (batch# A0375170), purchased from Acros Organic (Geel, Belgium); busulfan (batch# BCBN8120V), purchased from Sigma-Aldrich (Buchs, Switzerland); ganciclovir, purchased from Roche Pharma (Cymevene ® , batch# B4091B08, Reinach, Switzerland); cytarabine (batch# 3-YFD-59-1), epirubicin (batch# 11-CGS-118-1) and topotecan (batch# 8-MSW-162-1) purchased from Toronto Research Chemicals (North York, ON, Canada); raltitrexed (batch# MG10453-29102016), pemetrexed (batch# AGN2017-685), docetaxel (batch# CS13527-18112016), paclitaxel (batch# CS22539-20112016), vincristine sulfate (batch# MC-10452-0212016), doxorubicin hydrochloride (batch# MC10454-0612016), daunorubicin (batch# MC10456-1712016), idarubicin (batch# 5-CGS-96-1), etoposide (batch# MC10457-20102016), etoposide phosphate (batch# MC305547-21082017), irinotecan hydrochloride (batch# MC20783-21082017) and fludarabine phosphate (batch# MC444607-21082017) brought from Pharmaserv (Sansstad, Switzerland).

Techniques: