Biological Activity
| Description |
Lenalidomide (Revlimid, CC-5013) is a TNF-α secretion inhibitor with IC50 of 13 nM. |
| Targets |
TNF-α
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| IC50 |
13 nM [1]
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| In vitro |
Lenalidomide strongly induces IL-2 and sIL-2R production. Lenalidomide-induced tyrosine phosphorylation of CD28 on T cells is followed by a down-stream activation of NF-κB. [2] Lenalidomide and pomalidomide inhibits autoubiquitination of CRBN in HEK293 T cells expressing thalidomide-binding competent wild-type CRBN, but not thalidomide-binding defective CRBN(YW/AA). Overexpression of CRBN wild-type protein, but not CRBN(YW/AA) mutant protein, in KMS12 myeloma cells, amplifies pomalidomide-mediated reductions in c-myc and IRF4 expression and increases in p21(WAF-1) expression. Long-term selection for Lenalidomide resistance in H929 myeloma cell lines is accompanied by a reduction in CRBN, while in DF15R myeloma cells resistant to both pomalidomide and Lenalidomide, CRBN protein is undetectable. [3] Lenalidomide prevents induction of defects by down-regulating tumor cell inhibitory molecule expression. Lenalidomide prevents induction of tumor-induced T cell lytic synapse dysfunction. Lenalidomide treatment blocks CLL cell-induced T cell actin synapse dysfunction, mimicks antibody blockade, and down-regulates expression of CLL inhibitory ligands and their receptors on T cells. Lenalidomide treatment prevents tumor-induced immune suppression in FL, DLBCL, HL, MM, SCC, and OC and down-regulates immunosuppressive ligand expression on all tumor cells examined. CTL killing function significantly increases following antibody blockade of CLL inhibitory ligands or Lenalidomide treatment compared to control treatments. Treatment of autologous CLL-T cell co-cultures with Lenalidomide reverses impaired CD8+ T cell lytic synapse formation and granzyme B trafficking. [4] |
| In vivo |
The induction of angiogenesis by bFGF is significantly inhibited by oral treatment of Lenalidomide in a dose-dependent manner. Lenalidomide significantly decreases the percentage of vascularized area from 5.16% (control group) to 2.58% (50 mg/kg). Lenalidomide significantly reduces the calculated total MVL from 21.07 (control) to 8.11 (50 mg/kg). Lenalidomide significantly inhibites HUVEC migration through the fibronectin-coated membranes towards 0.1 ng/mL of bFGF at 100 μM, 1 ng/mL of VEGF at concentrations of 10 μM and 100 μM. [5] |
| Clinical Trials |
Lenalidomide has entered in a Phase II clinical trial in the treatment of chronic lymphocytic leukemia. |
| Features |
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Protocol(Only for Reference)
Kinase Assay:
[1]
| Assay for inhibition of TNF synthesis by human PBMCs |
Human PBMCs from normal donors are obtained by Ficoll−Hypaque density centrifugation. Cells (106 cells/mL) are cultured in RPMI supplemented with 10 AB+ serum, 2 mM l-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin. Lenalidomide is dissolved in DMSO at 20 mg/mL; further dilution is done with culture medium. The final DMSO concentration in all assays including the controls is 0.25%. Lenalidomide is added to cells 1 hour prior to the addition of LPS. PBMCs (106 cells/mL) are stimulated with 1 μg/mL of LPS from Salmonella minnesota R595. Cells, in triplicate, are incubated with LPS for 18−20 hours at 37 °C in 5% CO2. Supernatants are then harvested and assayed for cytokine levels. In some experiments, supernatants are kept frozen at −70 °C until use. Cell viability is assayed by Trypan blue exclusion dye method. The concentration of TNFα in the culture supernatants is determined by ELISA. Lenalidomide is assayed in a minimum of three separate experiments. Percent inhibition is determined as 100 × [1 − (cytokine(experimental)/cytokine(control))]. |
Animal Study:
[5]
| Animal Models |
Adult male Sprague-Dawley rats bearing HUVECs cells |
| Formulation |
0.5% DMSO |
| Dosages |
50 mg/kg and 250 mg/kg |
| Administration |
Administered via i.p. |
1
References
[1] Muller GW, et al. Bioorg Med Chem Lett, 1999, 9(11), 1625-1630.
[2] Zangari M, et al. Expert Opin Investig Drugs. 2005, 14(11), 1411-1418.
[3] Lopez-Girona A, et al. Leukemia. 2012.
[4] Ramsay AG, et al. Blood, 2012, 120(7), 1412-1421.
[5] Dredge K, et al. Microvasc Res. 2005, 69(1-2), 56-63.
[6] Henry JY, et al. Prostate. 2012, 72(8), 856-867.
[7] Ocio EM, et al. Haematologica, 2010, 95(5), 794-803.
[8] Henry JY, et al. J Clin Oncol, 2010, 28(suppl), Abst e13155.
[9] Moros A, et al. Canc Res, 2012, 72(8, Suppl 1), Abst 1942.
Download Lenalidomide (Revlimid) SDF
| Molecular Weight (MW) |
259.26 |
| Formula |
C13H13N3O3
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| CAS No. |
191732-72-6, 1243329-97-6 (HCl) |
| Synonyms |
CC-5013 |
Solubility (25°C)
- DMSO 52 mg/mL
- Water <1 mg/mL
- Ethanol <1 mg/mL
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| Storage |
2 years -20°CPowder |
| 2 weeks4°Cin DMSO |
| 6 months-80°Cin DMSO |
| Chemical Name |
3-(4-amino-1-oxoisoindolin-2-yl)piperidine-2,6-dione |
Research Area
Customer Reviews (4)
Click to enlarge
Effect of lenalidomide treatment (50 mg/kg/day, p.o. for 3 days) on expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), Fas, Fas ligand (FasL), and cleaved caspase-3 in myocardium from lean and ob/ob mice. (a) Representative gel blots of TNF-α, IL-6, Fas, FasL, cleaved caspase-3 and α-Tubulin (as loading control) using specific antibodies. (b) TNF-α.
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Rating |
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| Source |
Obesity (Silver Spring), 2012, 20(11), 2174-85. Lenalidomide (Revlimid) purchased from Selleck |
| Method |
Western Blot |
| Cell Lines |
ob/ob mice |
| Concentrations |
50 mg/kg/day |
| Incubation Time |
3 d |
| Results |
Although lenalidomide itself did not exert any effect on the expression of these proteins, it significantly attenuated or ablated obesity-induced upregulation of TNF-α, IL-6, Fas, and cleaved caspase-3 without any effect on FasL. |
Click to enlarge
(A) MM.1S cells were treated for 48h with indicated concentrations of MLN2238, lenalidomide, or MLN2238 plus lenalidomide; and then assessed for viability using MTT assays. Isobologram analysis shows the synergistic cytotoxic effect of MLN2238 and lenalidomide. The graph (left panel) is derived from the values given in the Table (right panel). The numbers 1–9 in graph represent combinations shown in the Table. Fa Com = Fraction of cells showing decrease in viability with MLN2238 plus lenalidomide treatment. Combination index (CI) of < 1 indicates synergy. All experiments were performed in triplicate and mean value is shown. (B) MM.1S cells were treated for 48h with indicated concentrations of MLN2238, SAHA, or MLN2238 plus SAHA; and then assessed for viability using MTT assays. Isobologram analysis shows the synergistic cytotoxic effect of MLN2238 and SAHA. The graph (left panel) is derived from the values given in the Table (right panel). The numbers 1–9 in graph represent combinations shown in the Table. Combination index (CI) of < 1 indicates synergy. (C) MM.1S cells were treated for 48h with indicated concentrations of MLN2238, Dex, or MLN2238 plus Dex; and then assessed for viability using MTT assays. Isobologram analysis shows the synergistic cytotoxic effect of MLN2238 and Dex. The graph (left panel) is derived from the values given in the Table (right panel).
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Rating |
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| Source |
Clin Cancer Res, 2011, 17(16), 5311-21. Lenalidomide (Revlimid) purchased from Selleck |
| Method |
MTT assays |
| Cell Lines |
MM.1S cells |
| Concentrations |
1-5 μM |
| Incubation Time |
48 h |
| Results |
The combination of low concentrations of MLN2238 and lenalidomide triggered synergistic anti-MM activity, with acombination index (CI) < 1.0. |
Click to enlarge
MM1S were treated with AT9283 (0.125 μM), lenalidomide (2 μM) or combined therapy for 72 hours. Annexin/PI staining show increased apoptosis associated with caspase 8 and PARP cleavage after 18 and 36 hours of exposure. B) MM.1S cells were treated with AT9283 (0.125 μM), lenalidomide (2 μM) or combined therapy for 4 hours. Whole lysates were immunoblotted with indicated antibodies
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Rating |
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| Source |
Harvard Medical School. Lenalidomide (Revlimid) purchased from Selleck |
| Method |
AnnexinV/PI staining, Western blotting |
| Cell Lines |
MM.1S cells |
| Concentrations |
2 μM |
| Incubation Time |
0-72 h |
| Results |
The analysis showed an increase (55.7%) of cells in early and late apoptosis after 72 hours of exposure to combined therapy. Combined AT9283 and lenalidomide increased cleavage of caspase-8 and PARP. Using western blot analysis to delineate the molecular mechanism underlying this combination, we found that combination treatment resulted in downregulation of pSTAT3 and pERK following 4 hours of treatment. |
Click to enlarge
MM.1S cells were cultured for 48 hours in the presence or absence of BMSCs with control media, AT9283, lenalidomide or AT9283 plus lenalidomide. Cell proliferation was assessed by 3H-TdR uptake (left). MM.1S cells were cultured in the absence or presence of BMSCs and treated for 4 hours with drugs alone or in combination. Whole lysates were immunoblotted with indicated antibodies.
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Rating |
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| Source |
Harvard Medical School. Lenalidomide (Revlimid) purchased from Selleck |
| Method |
Western blotting, Cell proliferation assay |
| Cell Lines |
MM.1S cells |
| Concentrations |
2 μM |
| Incubation Time |
4/48 h |
| Results |
Combined therapy inhibited 3H-TdR uptake of MM.1S cells cultured in the presence of BMSCs. Interestingly, consistent with this data, We observed that AT9283 plus lenalidomide downregulated the expression of the p-STAT3 and p-ERK in MM.1S cells cultured with BMSCs. |
Product Citations (8)
Potent in vitro and in vivo activity of an Fc-engineered humanized anti-HM1.24 antibody against multiple myeloma via augmented effector function. [Tai YT, et al. Blood 2012;119(9), 2074-2082]
PubMed: 22246035In vitro and in vivo selective antitumor activity of a novel orally bioavailable proteasome inhibitor MLN9708 against multiple myeloma cells. [Chauhan D, et al. Clin Cancer Res 2011;17(16):5311-21]
PubMed: 21724551Anti-myeloma activity of a multi targeted kinase inhibitor,AT9283, via potent Aurora Kinase and STAT3 inhibition either alone or in combination with lenalidomide. [Santo L, et al. Clin Cancer Res 2011;17, 3259-3271]
PubMed: 21430070
Poly(ADP-ribose) polymerase family member 14 (PARP14) is a novel effector of the JNK2-dependent pro-survival signal in multiple myeloma. [Barbarulo A, et al. Oncogene 2012;ahead of print]
PubMed: 23045269Expansion of polyfunctional HIV-specific T cells upon stimulation with mRNA electroporated dendritic cells in the presence of immunomodulatory drugs. [De Keersmaecker B, et al. J Virol 2012;86(17):9351-60]
PubMed: 22718823Halofuginone inhibits multiple myeloma growth in vitro and in vivo and enhances cytotoxicity of conventional and novel agents. [Leiba M, et al. Br J Haematol 2012;157(6):718-31]
PubMed: 22533681Short-Term Lenalidomide (Revlimid) Administration Ameliorates Cardiomyocyte Contractile Dysfunction in ob/ob Obese Mice. [Li L, et al. Obesity (Silver Spring) 2012;20(11):2174-85]
PubMed: 22522886Suppression of AKT Anti-Apoptotic Signaling by a Novel Drug Candidate Results in Growth Arrest and Apoptosis of Hepatocellular Carcinoma Cells. [Cuconati A, et al. PLoS One 2013;8(1), e54595]
PubMed: 23355882
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