Home JAK/STAT STAT inhibitor - DNA/RNA Synthesis inhibitor - Nucleoside Analog/Antimetabolite inhibitor - Apoptosis related activator Fludarabine For research use only.

Fludarabine

Synonyms: FaraA, Fludarabinum, NSC 118218

Fludarabine is a STAT1 activation inhibitor which causes a specific depletion of STAT1 protein (and mRNA) but not of other STATs. Also a DNA synthesis inhibitor in vascular smooth muscle cells. Fludarabine induces apoptosis.

Fludarabine Chemical Structure

Fludarabine Chemical Structure

CAS No. 21679-14-1

Purity & Quality Control

Fludarabine Related Products

Signaling Pathway

STAT Signaling Pathways

STAT Signaling Pathway

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
Jeko-1  Function Assay 20 μM 24 h inhibits expression of IDO 25940712
MV-4-11 Apoptosis Assay 2.5 μM 48 h induces apoptosis slightly 25111583
THP-1 Apoptosis Assay 2.5 μM 48 h induces apoptosis slightly 25111583
MOLM 13 Apoptosis Assay 2.5 μM 48 h induces apoptosis slightly 25111583
KBM3/Bu2506 Apoptosis Assay 2.5 μM 48 h induces apoptosis slightly 25111583
Nalm-6 Growth Inhibition Assay IC50=18 μM 25061101
Reh Growth Inhibition Assay IC50=30 μM 25061101
U2937 Growth Inhibition Assay IC50=16 μM 25061101
Mec-1 Growth Inhibition Assay IC50＞500 μM 25061101
RPMI-8226 Growth Inhibition Assay IC50=500 μM 25061101
Molt-4 Growth Inhibition Assay IC50=180 μM 25061101
Nalm-6-FluR Growth Inhibition Assay IC50=250 μM 25061101
Raji  Function Assay 3 μM 24/48/72 h induces accumulations of p53, p63 and p73  24940695
PBMC Function Assay 50/100 μM 24 h DMSO inhibits STAT1 phosphorylation 24911872
MDA-231 Function Assay 100 μM 24 h DMSO decreases IDO expression 24911872
624.38mel  Function Assay 50 μM 24 h DMSO decreases IDO expression 24911872
MDA-231 Function Assay 50-200 μM 24 h DMSO inhibits IDO activity independently of mRNA levels 24911872
624.38mel  Function Assay 50-200 μM 24 h DMSO inhibits IDO activity independently of mRNA levels 24911872
HMECs Function Assay 100 μM  36 h inhibits IFNγ and LPS induced STAT1 phosphorylation and IRF1 expression 24211327
HMECs  Function Assay 100 μM  36 h inhibits IFNα mediated phosphorylation of STAT1 and STAT3, but not of STAT2 24211327
BJAB Apoptosis Assay 5 μM 24 h induces cell apoptosis 24057147
I-83 Apoptosis Assay 5 μM 24 h induces cell apoptosis 24057147
NALM6 Apoptosis Assay 5 μM 24 h induces cell apoptosis 24057147
DU-145 Growth Inhibition Assay 0-10 μg/ml 48 h  inhibits cell growth in a dose-dependent manner 23734815
Nalm-6 Function Assay 10 μM 1/2/4 h induces autophagy 23681223
Reh Function Assay 10 μM 1/2/4 h induces autophagy 23681223
Nalm-6 Growth Inhibition Assay IC50 ∼10 μM 23681223
Reh Growth Inhibition Assay IC50 ∼10 μM 23681223
HEC1A Growth Inhibition Assay 100-500 μM 24 h inhibits cell growth in a dose-dependent manner 23595697
AN3CA Growth Inhibition Assay 100-500 μM 24 h inhibits cell growth in a dose-dependent manner 23595697
HEC50B Growth Inhibition Assay 100-500 μM 24 h inhibits cell growth slightly 23595697
HEC1A Apoptosis Assay 20/100 μM 24 h induces apoptosis in a dose-dependent manner 23595697
AN3CA Apoptosis Assay 20/100 μM 24 h induces apoptosis in a dose-dependent manner 23595697
HEC50B Apoptosis Assay 20/100 μM 24 h induces apoptosis slightly 23595697
EHEB Apoptosis Assay 40 μM 24 h induces apoptosis 23497075
A549 Growth Inhibition Assay IC50=15.7±2.8 µM 23377192
A549 GAPDH-deficient Growth Inhibition Assay IC50=18.5±2.3 µM 23377192
CLL  Apoptosis Assay 10 μM  24-96 h induces apoptotic cell death 22207686
MEC1 Apoptosis Assay 100 μM 72 h induces apoptosis significantly 22132973
U937  Apoptosis Assay 0.8 μM 4-48 h induces apoptosis slightly 22074700
U937  Apoptosis Assay 1 μM 96 h induces apoptosis slightly 22023523
Daudi Apoptosis Assay 20 μM 96 h induces apoptosis slightly 22023523
J45.01 Apoptosis Assay 1 μM 96 h induces apoptosis slightly 22023523
RPMI 8226 Growth Inhibition Assay IC50=25.9 ± 3.7 μM 21948264
CEM Growth Inhibition Assay IC50=2.4 ± 0.4 μM 21948264
Raji Growth Inhibition Assay IC50=0.47 ± 0.04 μM 21948264
U937 Growth Inhibition Assay IC50=0.24 ± 0.04 μM 21948264
K562 Growth Inhibition Assay IC50=0.44 ± 0.05 μM 21948264
NALM-6 Apoptosis Assay 10 μM  24 h induces cell apoptosis slightly 21699383
JMV-3 Apoptosis Assay 10 μM  24 h induces cell apoptosis slightly 21699383
EHEB Function Assay 5-50 μM 24 h decreases p21 expression significantly 21168391
JVM-2  Function Assay 30 μM 24 h decreases p21 expression 21168391
KBM3/Bu2506 Growth Inhibition Assay IC20=0.67 µM 20933509
KBM3/Bu2506 Growth Inhibition Assay 0.6 μM 24 h increases the cell fraction in S-phase 20933509
MDA-MB-231 Growth Inhibition Assay IC50=4.0 μM 20447390
MCF-7 Growth Inhibition Assay IC50=15.0 μM 20447390
HLE-B3  Function Assay 25 μM 48 h blocks IFN-γ–induced STAT1 phosphorylation and IDO expression 20435158
K562 Growth Inhibition Assay 72 h IC50=3.3 nM 20307198
BW-225 Growth Inhibition Assay IC20=1.37 ×10−8 μM  18661380
OH-65 Growth Inhibition Assay IC20=1.37 ×10−8 μM  18661380
GR-145 Growth Inhibition Assay IC20=2.74 × 10−8 μM  18661380
A549 Growth Inhibition Assay IC20=5.48 × 10−8 μM  18661380
CaSki  Growth Inhibition Assay IC20=1.37 × 10−7 μM  18661380
ZMK-1 Growth Inhibition Assay IC20=1.37 × 10−6 μM  18661380
SKW6.4 Apoptosis Assay 0.01-10 μM 24/48 h induces cell death in both time- and dose- dependent manner 18092340
RPMI 8226 Growth Inhibition Assay 24 h IC50=1.54 μM 17976186
MM.1S Growth Inhibition Assay 48 h IC50=13.48 μM 17976186
MM.1R Growth Inhibition Assay 48 h IC50=33.79 μM 17976186
U937 Growth Inhibition Assay IC50=3,200 ± 560 nM 15930361
CLL5 Antitumor assay 48 hrs Antitumor activity against CLL5 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 0.16 μM. 24673739
K562 Cytotoxicity assay 72 hrs Cytotoxicity against human paclitaxel-resistant K562 cells after 72 hrs by MTT assay, IC50 = 0.26 μM. 20605656
CLL3 Antitumor assay 48 hrs Antitumor activity against CLL3 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 0.35 μM. 24673739
CLL4 Antitumor assay 48 hrs Antitumor activity against CLL4 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 0.64 μM. 24673739
CEM-DNR-B Cytotoxicity assay 72 hrs Cytotoxicity against human CEM-DNR-B cells after 72 hrs by MTT assay, IC50 = 1.01 μM. 20605656
primary CLL Cytotoxicity assay Cytotoxicity against human primary CLL cells, LD50 = 1.1 μM. 25148392
CLL6 Antitumor assay 48 hrs Antitumor activity against CLL6 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 1.6 μM. 24673739
CLL2 Antitumor assay 48 hrs Antitumor activity against CLL2 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 2.66 μM. 24673739
HCT116 Cytotoxicity assay 72 hrs Cytotoxicity against human HCT116 cells after 72 hrs by SRB assay, IC50 = 6.6 μM. 25462277
PBMC Cytotoxicity assay 48 hrs Cytotoxicity against patient PBMC after 48 hrs by CellTitre-Blue assay in presenc of mouse M210B4 cells, IC50 = 10 μM. 25562417
CHO Function assay Binding affinity determined by displacement of specific binding of [125I]N-(4-amino-3-iodophenethyl)-adenosine in membranes of CHO cells stably transfected with the rat adenosine A3 receptor, Ki = 10.4 μM. 7707320
JVM2 Antitumor assay Antitumor activity against human JVM2 cells assessed as cell viability after 48 hrs by FACS analysis, EC50 = 10.4 μM. 24673739
HeLa Antitumor assay Antitumor activity against human HeLa cells assessed as cell viability by MTT assay, EC50 = 16 μM. 24673739
CLL1 Antitumor assay 48 hrs Antitumor activity against CLL1 cells isolated from CLL patient assessed as cell viability after 48 hrs by FACS analysis, EC50 = 17.1 μM. 24673739
CEM Cytotoxicity assay 72 hrs Cytotoxicity against human CEM cells after 72 hrs by MTT assay, IC50 = 19.49 μM. 20605656
T47D Cytotoxicity assay 72 hrs Cytotoxicity against human T47D cells after 72 hrs by SRB assay, IC50 = 46.2 μM. 25462277
A549 Cytotoxicity assay 72 hrs Cytotoxicity against human A549 cells after 72 hrs by MTT assay, IC50 = 47.44 μM. 20605656
CCRF-CEM Function assay 10 uM 1 to 60 mins Drug transport in human CCRF-CEM cells assessed as ENT1-mediated uptake at 10 uM after 1 to 60 mins by liquid scintillation counting analysis 23388705
TC32 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for TC32 cells 29435139
U-2 OS qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells 29435139
DAOY qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells 29435139
Saos-2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) cells 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells 29435139
BT-12 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for LAN-5 cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for NB-EBc1 cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells 29435139
Rh41 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells 29435139
A673 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells) 29435139
BT-37 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-37 cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for MG 63 (6-TG R) cells 29435139
Rh30 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh30 cells 29435139
U-2 OS qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for U-2 OS cells 29435139
OHS-50 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for OHS-50 cells 29435139
SK-N-SH qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for SK-N-SH cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for RD cells 29435139
Daoy qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for Daoy cells 29435139
TC32 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D caspase screen for TC32 cells 29435139
TC32 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for TC32 cells 29435139
MG 63 (6-TG R) qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for MG 63 (6-TG R) cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells 29435139
NB-EBc1 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells 29435139
LAN-5 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells 29435139
NB1643 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for NB1643 cells 29435139
SK-N-MC qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-MC cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells 29435139
TC32 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for TC32 cells 29435139
Rh18 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh18 cells 29435139
Saos-2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Saos-2 cells 29435139
SJ-GBM2 qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for SJ-GBM2 cells 29435139
RD qHTS assay qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for RD cells 29435139
Click to View More Cell Line Experimental Data

Biological Activity

Description Fludarabine is a STAT1 activation inhibitor which causes a specific depletion of STAT1 protein (and mRNA) but not of other STATs. Also a DNA synthesis inhibitor in vascular smooth muscle cells. Fludarabine induces apoptosis.
Targets
STAT1 [4]
(Vascular smooth muscle cells)
In vitro
In vitro

Fludarabine efficiently inhibits the proliferation of RPMI 8226 cells with IC50 of 1.54 μg/mL. The IC50 of Fludarabine against MM.1S and MM.1R cells is 13.48 μg/mL and 33.79 μg/mL, respectively. In contrast, U266 cells are resistant to Fludarabine with IC50 of 222.2 μg/mL. Fludarabine treatment results in increased number of cells in the G1 phase of cell cycle, accompanied with a concomitant reduction of cells at the S phase of cell cycle in a time-dependent manner. Fludarabine induces a cell cycle block and triggers apoptosis in MM cells. Fludarabine triggers time-dependent cleavage of caspase-8, -9, and -3, -7, followed by PARP cleavage. Fludarabine increases expression of Bax in a time-dependent fashion, while the expression of Bak doesn't change. After exposure to Fludarabine for 12 hours, RPMI 8226 cells shows a loss of membrane potential with 61.05% of the cells expressing low fluorescence of rhodamine 123 compared with 8.62% of cells in untreated control. [1] To enhance solubility, Fludarabine is formulated as the monophosphate (F-ara-AMP, fudarabine), which is instantaneously and quantitatively dephosphorylated to the parent nucleoside upon intravenous infusion. Inside the cells rephosphorylation occurs which leads to fuoroadenine arabinoside triphosphate (F-ara-ATP), the major cytotoxic metabolite of F-ara-A. [2] Fludarabine can also induce pro-inflammatory stimulation of monocytic cells, as evaluated by increased expression of ICAM-1 and IL-8 release. [3] Fludarabine does not affect the growth of ovarian cancer cell lines, whereas it induces marked and dose-dependent inhibition of proliferation in melanoma cell lines. [4] Fludarabine is an inhibitor of STAT1 that specifically reduces STAT1 without affecting other STAT family members[5]. In addition to cytoplasmic accumulation, repeated low-dose cisplatin (RLDC) induces HMGB1 expression, which is marked suppressed by STAT1 knockdown. Consistently, fludarabine suppresses HMGB1 expression during RLDC treatment dose-dependently in RLDC-treat renal tubular cells[5].
Cell Research Cell lines Dexamethasone-sensitive (MM.1S) and -resistant (MM.1R) human MM cell lines, RPMI8226 and U266 cell lines
Concentrations 2 μg/mL
Incubation Time 24 hours
Method

After treated with Fludarabine or control, dexamethasone-sensitive (MM.1S) and -resistant (MM.1R) human MM cell lines, RPMI8226 and U266 cell lines (5 × 105 cells) are washed twice in phosphate-buffered saline (PBS) and fixed with 70% ice-cold ethanol, then centrifuged and suspended in PBS containing 100 μg/mL RNase A. After incubated for 30 minutes at 37 ºC, samples are resuspended in 25 μg/mL propidium iodide. Flow cytometry is performed on a FACSCalibur automated system. Apoptosis is determined by Annexin V-FITC apoptosis detection kit, according to the manufacturer's instructions. For TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) assay, cells are analyzed by flow cytometry using the in situ cell death detection kit.
Experimental Result Images Methods Biomarkers Images PMID
Western blot STAT1 p-p53 / p53 procaspase-9 / procaspase-3 26677135
Immunofluorescence α-SMA / Vimentin 28322315
Growth inhibition assay Cell viability 24956101
In Vivo
In vivo

Tumors treated with PBS grow rapidly to approx-imately 10-fold their initial volume in 25 day, whereas, the tumors in the Fludarabine at 40 mg/kg increase less than 5-fold. A significant antitumor effect of 40 mg/kg Fludarabine on RPMI8226 tumor growth is demonstrated. RPMI8226 tumors treated with 40 mg/kg Fludarabine at day 10 increase apoptotic nuclei. Fludarabine is effective in suppressing RPMI8226 myeloma xenografts in SCID mice. [1]
Animal Research Animal Models Severe combined immunodeficient (SCID) mice bearing RPMI 8226 cells
Dosages 40 mg/kg
Administration Administered via i.p.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT05390814 Recruiting
Primary Central Nervous System Lymphoma
Assistance Publique - Hôpitaux de Paris
 December 18 2023 Phase 1
NCT05201183 Withdrawn
Acute Myeloid Leukemia|Chronic Myeloid Leukemia|Acute Lymphocytic Leukemia|Myelodysplastic Syndromes
Naoyuki G. Saito M.D. Ph.D.|Indiana University
 October 2023 Phase 1|Phase 2
NCT05917405 Recruiting
Acute Myeloid Leukemia in Remission
Nantes University Hospital
 September 14 2023 Phase 2

Chemical Information & Solubility

Molecular Weight 285.23 Formula

C10H12FN5O4
CAS No. 21679-14-1 SDF Download Fludarabine SDF
Smiles C1=NC2=C(N=C(N=C2N1C3C(C(C(O3)CO)O)O)F)N
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 57 mg/mL ( (199.83 mM) Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Water : Insoluble

Ethanol : Insoluble


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In vivo
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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
how to re-suspend and deliver the inhibitor for in vivo experiments?

Answer:
For S1491, Fludarabine, we tested a vehicle: 30% Propylene glycol, 5% Tween 80, 65% D5W that you can resuspend the compound in at up to 30mg/ml. It's a suspension and can only be given via oral gavage.

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