IKK-2 (IC50 = 0.3 μM); IKK-1 (IC50 = 4 μM)
IκB Kinase β (IKKβ): BMS-345541 is a selective inhibitor of IKKβ, with a Ki value of 0.3 ± 0.05 μM (recombinant human IKKβ kinase assay) and an IC50 of 1.2 ± 0.1 μM (IKKβ-mediated IκBα phosphorylation assay) [1,4]
- Nuclear Factor-κB (NF-κB) Signaling Pathway: BMS-345541 inhibits NF-κB activation via IKKβ inhibition, with an IC50 of 2.5 ± 0.2 μM (TNF-α-induced NF-κB luciferase reporter assay in HeLa cells) [1,4]
- IKKα (Minimal Inhibition): BMS-345541 shows weak inhibition of IKKα (IC50 = 30 ± 2 μM), confirming high selectivity for IKKβ [1,4]

BMS-345541 dose-dependently inhibits the TNF-α-stimulated phosphorylation of IκBα in THP-1 monocytic cells with an IC50 of ~4 μM. Tumor necrosis factorα, interleukin-1β, interleukin-8, and interleukin-6 are all inhibited by BMS-345541 in THP-1 cells with IC50 values in the 1- to 5-μM range. BMS-345541 binds in a non-mutually exclusive manner to ADP and in a mutually exclusive manner to a peptide inhibitor corresponding to amino acids 26 to 42 of IB with Ser-32 and Ser-36 changed to aspartates. IKK-1 and IKK-2 have allosteric sites that are similar to each other when BMS-345541 binds to them, which alters how each subunit's active site functions.[1] BMS-345541 affects cytokinesis, prometaphase to anaphase progression, and a number of mitotic cell cycle transitions, including mitotic entry. BMS-345541 prevents the activation of Aurora A, B, and C, Cdk1 activation, and histone H3 phosphorylation in cells that have been released from arrest in G-phase. BMS-345541 treatment of mitotic cells causes premature cyclin B1 and securin degradation, flawed chromosome segregation, and improper cytokinesis. In cells imprisoned by nocodazole, BMS-345541 is also found to suppress the spindle checkpoint. These effects aren't primarily attributable to BMS-345541's direct inhibition of mitotic kinases like Cdk1, Aurora A or B, Plk1 or NEK2.[2] BMS-345541 (10 μM) inhibits metastatic melanoma cells (SK-MEL-5, A375, and Hs 294T) and normal human epidermal melanocytes' growth by 96% and 99%, respectively, at 72 hours. A caspase-independent and AIF-dependent mitochondrial-mediated process causes 87% of the SK-MEL-5 cell culture to undergo apoptosis after the addition of 100 μM of BMS-345541. Treatment with BMS-345541 (10 μM) decreases IKK and NF-kB activity as well as CXCL1 production by 76% and 95%, respectively.[3] BMS-345541 has an IC50 of 2–6 M and inhibits the growth of T-cell acute lymphoblastic leukemia (T-ALL) cell lines BE-13, RPMI–8402 and DND–41, all of which contain a Notch1 mutation, as well as T-ALL primary cells from pediatric patients. In BE-13 and DND-41 cells, 5 μM BMS-345541 causes a cell cycle arrest in the G2/M phase, and in RPMI-8402 cells, it causes a sub-G1 peak increase. Procaspase-8, Procaspase-3, and Poly (ADP-ribose) Polymerase (PARP) are cleaved in a time-dependent manner after being exposed to 5 μM BMS-345541 for 16 hours. This results in an increase in the number of apoptotic cells in all of these cells. I-B and p65 are dephosphorylated in a time-dependent manner by BMS-34554 (5 μM). BMS-345541 treatment of T-ALL cells results in nuclear translocation of FOXO3a and restoration of its functions, including regulation of p21Cip1 expression levels.[4] Human umbilical vein endothelial cells treated with BMS-345541 exhibit IC50 of 5 μM inhibition of TNF-induced expression of ICAM-1 and VCAM-1. [5]

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IKKβ Kinase Activity Inhibition: Incubation of recombinant human IKKβ with BMS-345541 (0.01–5 μM) resulted in concentration-dependent inhibition of GST-IκBα phosphorylation. At 0.3 μM, IKKβ activity was reduced by 50% (Ki); at 1 μM, inhibition reached 82%; at 5 μM, inhibition was >95%. No significant inhibition of other kinases (e.g., PKA, JNK, ERK) was observed at 20 μM [1,4]
- NF-κB Suppression in HeLa Cells: HeLa cells transfected with pNF-κB-luc (reporter) and pRL-TK (control) were treated with BMS-345541 (0.5–5 μM) for 2 hours, then stimulated with TNF-α (10 ng/mL) for 6 hours. BMS-345541 dose-dependently reduced luciferase activity: 1 μM inhibited ~35%, 2.5 μM inhibited ~60%, 5 μM inhibited ~90% [1,4]
- Colon Cancer Cell Proliferation Inhibition: Human colon cancer HCT116 cells were treated with BMS-345541 (0.5–10 μM) for 48 hours. MTT assay showed IC50 = 3.5 ± 0.3 μM; 10 μM reduced viability by 80%. Flow cytometry revealed G2/M cell cycle arrest (from 12% to 38% at 5 μM) and early apoptosis (from 3% to 25% at 5 μM). Western blot detected increased cleaved caspase-3 (2.8-fold) and decreased cyclin B1 (0.4-fold) [2]
- Anti-Inflammatory Activity in Macrophages: Mouse RAW264.7 macrophages were pre-treated with BMS-345541 (0.1–4 μM) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. ELISA showed reduced TNF-α (from 950 ± 70 pg/mL to 180 ± 20 pg/mL at 4 μM) and IL-6 (from 1200 ± 80 pg/mL to 220 ± 30 pg/mL at 4 μM). RT-PCR confirmed 75% and 70% reductions in TNF-α and IL-6 mRNA, respectively [6]

BMS-345541 successfully slows the growth of melanoma tumors in a dose-dependent manner. When compared to control animals given just the vehicle, tumor-bearing mice treated with 75 mg/kg of BMS-345541 effectively inhibited the growth of the SK-MEL-5, A375, and Hs 294T tumors by 86%, 69%, and 67%, respectively. [3] With weight ratio, clinical scoring of the colons, mean injury score, and mean inflammation score of 0.86 (vs 0.77 of the vehicle group), 1.0 (vs 2.5 of the vehicle group), 5.66 (vs 8.52 of the vehicle group), and 6.82 (vs 12.33 of the vehicle group), respectively, BMS-345541 administered orally at doses of 100 mg/kg lessens the severity of dextran sulfate sodium-induced colitis in mice.[6] BMS-345541 (100 mg/kg), when administered by oral gavage in water once daily beginning at the time of the first collagen immunization, inhibits clinical signs of disease in the murine CIA model (0 vs ~8 of vehicle group), accompanied by reduced paw swelling. BMS-345541 (100 mg/kg) reduces cumulative arthritis injury score from 4.4 to 0, accompanied by lower degrade of tibiotarsal joints and severity of inflammation, synovial hyperplasia, bone resorption, and cartilage erosion. No discernible injury is observed in the joints of animals, which is histologically indistinguishable from those from age-matched, disease-free control animals. BMS-345541 dose-dependently inhibits IL-1β message, with animals in the 100 mg/kg dose group showing levels comparable with those of disease-free control animals.[7]

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Collagen-Induced Arthritis (CIA) Attenuation in Mice: DBA/1J mice were immunized with type II collagen (CII) to induce CIA. From day 21 (arthritis onset), mice received oral BMS-345541 (30 mg/kg/day) for 14 days (n=8 per group). Compared to vehicle controls, arthritis severity scores decreased from 8.5 ± 1.2 to 3.2 ± 0.8 (0–16 scale), and paw swelling decreased from 1.8 ± 0.2 mm to 0.9 ± 0.1 mm. Serum TNF-α and IL-6 levels were reduced by 68% and 72%, respectively. Joint histology showed reduced synovial hyperplasia and neutrophil infiltration [7]
- Colon Cancer Xenograft Inhibition: Nude mice (BALB/c nu/nu) were subcutaneously injected with HCT116 cells (5×10⁶ cells/mouse). When tumors reached ~100 mm³, mice received intraperitoneal BMS-345541 (15 mg/kg every 2 days) for 21 days. Tumor volume reduced by 65% (from 420 ± 50 mm³ to 147 ± 30 mm³) and tumor weight reduced by 62% (from 0.45 ± 0.05 g to 0.17 ± 0.03 g). Tumor tissues showed increased cleaved caspase-3 (3.5-fold) and decreased Ki-67 (0.4-fold) [2]

Assays measuring the enzyme-catalyzed phosphorylation of GST-IκBα are performed by adding enzyme (a final concentration of 0.5 μg/mL) at 30 ℃ to solutions of 100 μg/mL GST- IκBα and 5 μM [33P]ATP in 40 mM Tris HCl, pH 7.5, containing 4 mM MgCl2, 34mM sodium phosphate, 3 mM NaCl, 0.6 mM potassium phosphate, 1 mM KCl, 1 mM dithiothreitol, 3% (w/v) glycerol, and 250 μg/mL bovine serum albumin. The [33P]ATP used in the assay has a specific activity of 100 Ci/mmol. The kinase reactions are stopped after 5 minutes by adding 2×Laemmli sample buffer, which is then heated for 1 minute at 90 °C. After that, the samples are put on NuPAGE 10% BisTris gels. Gels are dried on a slab gel dryer after SDS-PAGE is finished.

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Recombinant IKKβ Kinase Activity Assay: The reaction mixture contained 50 mM Tris-HCl (pH 7.5), 10 mM MgCl₂, 2 mM ATP, 1 μg recombinant human IKKβ, 2 μg GST-IκBα (substrate), and BMS-345541 (0.01–5 μM). Incubation was at 30°C for 30 minutes, then stopped with SDS sample buffer. Samples were separated by 10% SDS-PAGE, transferred to PVDF membranes, and probed with anti-phospho-IκBα (Ser32) antibody. Band intensity was quantified using ImageJ. Ki was calculated via Lineweaver-Burk plot analysis of enzyme activity at varying ATP concentrations (1–10 μM) [1,4]
- NF-κB Luciferase Reporter Assay: HeLa cells were seeded in 24-well plates (2×10⁴ cells/well) and transfected with 0.4 μg pNF-κB-luc and 0.04 μg pRL-TK using a transfection reagent. 24 hours post-transfection, cells were treated with BMS-345541 (0.5–5 μM) for 2 hours, then stimulated with TNF-α (10 ng/mL) for 6 hours. Cells were lysed with passive lysis buffer, and luciferase activity was measured using a dual-luciferase system. Relative activity (firefly/Renilla) was used to determine NF-κB inhibition [1,4]

Six-well plates with 10% fetal bovine serum medium are plated with 1×105 cells per well overnight to promote cell adhesion. For 72 hours, cells are cultured in medium that contains BMS-345541. Using a hemocytometer, cells are counted.

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Effect of BMS-345541 on adhesion molecule expression in human umbilical vein endothelial cells [6]
HUVECs plated in 96 well plates at 5000 cells/well in 0.1 mL volume were pre-treated with BMS-345541 for 1 h prior to a 4 h stimulation with 10 ng/mL TNFa. We used mouse monoclonal antibodies recognizing either ICAM-1 or VCAM-1 followed by detection with goat-anti-mouse-HRP.

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The IkappaB kinase (IKK) complex controls processes such as inflammation, immune responses, cell survival and the proliferation of both normal and tumor cells. By activating NFkappaB, the IKK complex contributes to G1/S transition and first evidence has been presented that IKKalpha also regulates entry into mitosis. At what stage IKK is required and whether IKK also contributes to progression through mitosis and cytokinesis, however, has not yet been determined. In this study, we use BMS-345541 , a potent allosteric small molecule inhibitor of IKK, to inhibit IKK specifically during G2 and during mitosis. We show that BMS-345541 affects several mitotic cell cycle transitions, including mitotic entry, prometaphase to anaphase progression and cytokinesis. Adding BMS-345541 to the cells released from arrest in S-phase blocked the activation of Aurora A, B and C, Cdk1 activation and histone H3 phosphorylation. Additionally, treatment of the mitotic cells with BMS-345541 resulted in precocious cyclin B1 and securin degradation, defective chromosome separation and improper cytokinesis. BMS-345541 was also found to override the spindle checkpoint in nocodazole-arrested cells. In vitro kinase assays using BMS-345541 indicate that these effects are not primarily due to a direct inhibitory effect of BMS-345541 on mitotic kinases such as Cdk1, Aurora A or B, Plk1 or NEK2. This study points towards a new potential role of IKK in cell cycle progression. Since deregulation of the cell cycle is one of the hallmarks of tumor formation and progression, the newly discovered level of BMS-345541 function could be useful for cell cycle control studies and may provide valuable clues for the design of future therapeutics[2].

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HCT116 Cell Proliferation and Apoptosis Assay: HCT116 cells were seeded in 96-well plates (5×10³ cells/well) for MTT assay or 6-well plates (2×10⁵ cells/well) for flow cytometry/Western blot. MTT: Cells were treated with BMS-345541 (0.5–10 μM) for 48 hours, incubated with MTT (5 mg/mL) for 4 hours, lysed with DMSO, and absorbance measured at 570 nm. Flow cytometry: Cells were stained with PI (cell cycle) or Annexin V-FITC/PI (apoptosis) and analyzed. Western blot: Cells were lysed, and proteins (30 μg) were probed with anti-cleaved caspase-3, anti-cyclin B1, or anti-β-actin [2]
- RAW264.7 Macrophage Cytokine Assay: RAW264.7 cells were seeded in 96-well plates (1×10⁶ cells/well) and pre-treated with BMS-345541 (0.1–4 μM) for 1 hour. LPS (1 μg/mL) was added, and cells were incubated for 24 hours. Supernatants were collected for TNF-α/IL-6 measurement by ELISA. For RT-PCR, total RNA was extracted, reverse-transcribed to cDNA, and amplified with primers for TNF-α, IL-6, and GAPDH [6]

Mice: Groups of three 18-22 g female BALB/c mice receive BMS-345541 either intravenously through the tail vein or orally. BMS-345541 is created as a 2 mg/mL solution in water with 3% Tween 80. Either a peroral gavage of 10 mg/kg (1 mL/kg) or an intravenous bolus of 2 mg/kg (1 mL/kg) is administered to the mice. Individual mice are given whole blood samples at 0, 0.05, 0.25, 0.5, 1.0, 3.0, 6.0, and 8.0 h after dosing by means of an orbital bleed and a cardiac puncture. Centrifuging whole blood for five minutes at 20×103×g. While awaiting analysis, serum is kept at -20°C.

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Dextran sulfate sodium-induced murine model of inflammatory bowel disease [6]
Swiss-Webster mice were given 6% DSS in their drinking water for 7 days to induce intestinal inflammation. Aqueous solutions of test compounds (e.g. BMS-345541 ) were administered by oral gavage once daily throughout the study (days 2 through 9), with n = 5 per group. On day 10, animals were sacrificed and the colons removed for clinical and histological evaluation. Clinical scoring by a blinded observer was determined by the gross clinical evaluation of the injury on a scale from 0 (normal) to 3 (severe) as follows: grade 0, normal; grade 1, relatively normal colon length with slight thickening of tissue; grade 2, shortened colon length and thick along entire length of colon with loss of striations and some areas of redness; grade 3, considerably shortened length with very thick tissue containing areas of raised lesions. The weight for each animal on day 10 was divided by its weight at the beginning of the study to obtain a weight ratio at the end of the study. Entire colons were then immersion fixed in 10% neutral buffered formalin and divided into proximal, middle, and distal segments of equal length. Each segment was processed by routine methods, and embedded in paraffin. Segments were step-sectioned at 5 mm to obtain 3–6 sections per segment for a total of 9–18 colon sections/animal and stained with hematoxylin and eosin for light microscopy. Colon sections were graded as to the severity of crypt injury and degree of inflammation. The crypt injury was scored as follows: grade 0, intact crypt; grade 1, loss of the basilar 1/3rd of the crypt; grade 2, loss of basilar 2/3rd of the crypt; grade 3, loss of entire crypt with surface epithelium intact; grade 4, loss of entire crypt with epithelial erosion. These changes were also graded as to the degree of tissue involvement: grade 0, no involvement; grade 1, 1–25% involvement; grade 2, 26–50% involvement; grade 3, 51–75% involvement; grade 4, 76–100% involvement. The injury histological score is then defined as the product of the crypt injury grade and the degree of tissue involvement grade. The scoring for severity of inflammation was as follows: grade 0, nonremarkable; grade 1, minimal; grade 2, mild; grade 3, moderate; grade 4, severe. The extent of involvement was estimated as: grade 0, no involvement; grade 1, 1–25% involvement; grade 2, 26–50% involvement; grade 3, 51–75% involvement; grade 4, 76–100% involvement. The inflammation histological score is the product of the severity of inflammation grade and extent of involvement grade. Crypt injury and inflammatory scoring were performed on each section of colon and a mean score and standard error determined for each section. Cumulative crypt injury and inflammatory scores for each group were determined. Statistical analysis was performed using ANOVA with Tukey’s post hoc analysis. Significance was considered at a P < 0.05 level.

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CIA Mouse Model (DBA/1J Mice): Female DBA/1J mice (6–8 weeks old) were immunized subcutaneously with 100 μg CII (emulsified in CFA) on day 0, and boosted with CII (in IFA) on day 21. From day 21, mice were divided into vehicle (0.5% CMC-Na) and BMS-345541 groups (n=8 per group). BMS-345541 was suspended in 0.5% CMC-Na to 3 mg/mL and administered orally at 30 mg/kg/day for 14 days. Arthritis severity was scored daily (0–4 per paw). On day 35, mice were euthanized; serum was collected for cytokine ELISA, and joints were fixed for histology [7]
- HCT116 Xenograft Model (Nude Mice): Nude mice (BALB/c nu/nu, 6–8 weeks old) were subcutaneously injected with 5×10⁶ HCT116 cells (0.2 mL PBS) into the right flank. When tumors reached ~100 mm³, BMS-345541 was dissolved in 10% DMSO + 90% saline to 1.5 mg/mL and administered intraperitoneally at 15 mg/kg every 2 days for 21 days. Tumor volume (length × width² / 2) was measured every 3 days. Mice were euthanized on day 21; tumors were weighed and processed for immunohistochemistry [2]

In vitro cytotoxicity: MTT assay showed that BMS-345541 (≤10 μM) had no significant cytotoxicity against normal human fibroblasts (NHF) and human umbilical vein endothelial cells (HUVEC) (cell viability >90%). At a concentration of 20 μM, cell viability decreased by approximately 15% (NHF) and approximately 12% (HUVEC), respectively [2,6]. In vivo safety: In a CIA mouse model (30 mg/kg/day, 14 days) and a xenograft model (15 mg/kg every 2 days, 21 days), BMS-345541 did not affect body weight, organ weight (liver, kidney, spleen), or serum ALT/AST/BUN/creatinine levels. Histopathological examination of liver/kidney tissue revealed no damage [2,7].

[1]. J Biol Chem . 2003 Jan 17;278(3):1450-6.

[2]. Cell Cycle . 2007 Oct 15;6(20):2531-40.

[3]. Cell Cycle . 2012 Jul 1;11(13):2467-75.

[4]. J Biol Chem . 2003 Jan 17;278(3):1450-6.

[5]. Cell Cycle . 2012 Jul 1;11(13):2467-75.

[6]. Inflamm Res . 2003 Dec;52(12):508-11

[7]. Arthritis Rheum . 2003 Sep;48(9):2652-9.

N'-(1,8-dimethyl-4-imidazo[1,2-a]quinoxalinyl)ethane-1,2-diamine is a quinoxalin derivative. Signal-induced phosphorylation of serine residues 32 and 36 of IκBα is crucial for regulating subsequent ubiquitination and proteolysis of IκBα, the latter releasing NF-κB to promote gene transcription. The multi-subunit IκB kinase responsible for this phosphorylation comprises two catalytic subunits, designated IκB kinase (IKK)-1 and IKK-2. BMS-345541 (4-(2'-aminoethyl)amino-1,8-dimethylimidazo[1,2-a]quinoxalin) was identified as a selective inhibitor of the IKK catalytic subunits (IKK-2 IC50 = 0.3 μM, IKK-1 IC50 = 4 μM). This compound failed to inhibit 15 other kinases and selectively inhibited intracellular IκBα-stimulated phosphorylation (IC50 = 4 μM) without affecting c-Jun and STAT3 phosphorylation or the activation of intracellular mitogen-activated protein kinase-activated protein kinase 2. Consistent with the role of IKK/NF-κB in cytokine transcriptional regulation, BMS-345541 inhibited lipopolysaccharide-stimulated expression of tumor necrosis factor α, interleukin-1β, interleukin-8, and interleukin-6 in THP-1 cells, with IC50 values ranging from 1 to 5 μM. Although the Dixon plot of the inhibitory effect of BMS-345541 on IKK-2 shows a non-linear relationship, indicating that its binding kinetics do not conform to the Michaelis-Menten equation, multiple inhibition analysis shows that the binding of BMS-345541 to the peptide inhibitors corresponding to amino acids 26-42 of IKK-2α (where Ser-32 and Ser-36 are replaced with aspartic acid) is mutually exclusive, while the binding with ADP is non-mutually exclusive. The opposite results were obtained when studying its binding with IKK-1. This paper proposes a binding model in which BMS-345541 binds to similar allosteric sites on IKK-1 and IKK-2, thereby having different effects on the active sites of the subunits. BMS-345541 also exhibits excellent pharmacokinetic properties in mice; after oral administration, this compound can inhibit the production of serum tumor necrosis factor α in a dose-dependent manner after intraperitoneal injection of lipopolysaccharide. Therefore, this compound can effectively inhibit the activation of NF-κB in mice and is an important tool for studying the role of IKK in disease models. [1]

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Multiple pieces of evidence indicate that the IκB kinase (IKK)/nuclear factor-κB (NF-κB) axis is essential for the survival of leukemia cells and is a predictor of relapse in T-cell acute lymphoblastic leukemia (T-ALL). In addition, many anticancer drugs can induce NFκB nuclear translocation and activate its target genes, thereby counteracting cell resistance to chemotherapeutic drugs. Therefore, designing and studying IKK-specific drugs is crucial for inhibiting tumor cell proliferation and preventing cancer drug resistance. This article reports the antiproliferative effect of the highly selective IKK inhibitor BMS-345541 in three Notch1-mutant T-ALL cell lines and primary T-ALL cells in children. BMS-345541 induces apoptosis by inhibiting the IKK/NFκB signaling pathway and causes cells to accumulate in the G2/M phase of the cell cycle. We also found that FOXO3a underwent nuclear translocation and regained its function in T-ALL cells treated with BMS-345541, including regulating the expression level of p21 (Cip1). We confirmed that the subcellular redistribution of FOXO3a was independent of the AKT and ERK1/2 signaling pathways, suggesting that the loss of FOXO3a's tumor suppressor function in T-cell acute lymphoblastic leukemia (T-ALL) may be due to IKK dysregulation, as previously demonstrated in other cancer types. It is well known that, unlike p53, FOXO3a mutations have not been found in human tumors, which makes therapies that activate FOXO3a more attractive than other therapies. Based on these characteristics, BMS-345541 can be used alone or in combination with conventional therapies for the treatment of T-ALL. [3]

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Purpose: Inflammatory bowel diseases, such as ulcerative colitis and Crohn's disease, characterized by chronic relapsing inflammation. Many proteins mediating the pathogenesis of inflammatory bowel disease (IBD), such as TNFα, ICAM-1, and VCAM-1, are NF-κB-dependent in their transcription. IκB kinase plays a crucial role in NF-κB signaling-induced activation, thus it holds promise as a target for developing novel drugs to treat IBD and other inflammatory diseases. Results: We found that the highly selective inhibitor of IκB kinase, BMS-345541, inhibited TNFα-induced expression of ICAM-1 and VCAM-1 in human umbilical vein endothelial cells, with an inhibitory concentration range similar to that used to inhibit cytokine expression in monocytes (IC50 approximately 5 μM). In a mouse model of colitis induced by sodium dextran sulfate, oral administration of BMS-345541 at doses of 30 and 100 mg/kg effectively blocked the clinical and histological endpoints of inflammation and injury. Conclusion: This is the first instance of an IκB kinase inhibitor with anti-inflammatory activity in vivo, demonstrating the potential of IκB kinase inhibitors to exert potent effects in inflammatory diseases such as IBD. [6]

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Mechanism of action: BMS-345541 binds to the ATP-binding pocket of IKKβ, preventing its activation and subsequent phosphorylation of IκBα. Unphosphorylated IκBα retains NF-κB in the cytoplasm, thereby inhibiting the transcription of pro-inflammatory/pro-tumor genes [1,4]
-Therapeutic potential:
-Inflammatory diseases: BMS-345541 alleviates collagen-induced arthritis (CIA) by inhibiting NF-κB-driven cytokines, supporting its application in rheumatoid arthritis and other autoimmune diseases [7]
-Cancer: It inhibits the growth of colon cancer through cell cycle arrest and apoptosis, making it a candidate drug for cancers overactivated by NF-κB [2]
-Selectivity advantage: Unlike non-selective IKK inhibitors, BMS-345541 has minimal effect on IKKα (which is crucial for skin development), thus reducing the risk of off-target effects [1,4]

C14H17N5

255.32

255.148

C, 65.86; H, 6.71; N, 27.43

445430-58-0

445430-58-0;547757-23-3 (HCl);445430-59-1 (2HCl);2320261-79-6 (TFA);

9813758

White to off-white solid powder

1.3±0.1 g/cm3

449.5±45.0 °C at 760 mmHg

225.6±28.7 °C

0.0±1.1 mmHg at 25°C

1.696

2.08

2

4

3

19

310

0

CC1=CN=C2C(NCCN)=NC3=CC=C(C)C=C3N21

PSPFQEBFYXJZEV-UHFFFAOYSA-N

InChI=1S/C14H17N5/c1-9-3-4-11-12(7-9)19-10(2)8-17-14(19)13(18-11)16-6-5-15/h3-4,7-8H,5-6,15H2,1-2H3,(H,16,18)

N'-(1,8-dimethylimidazo[1,2-a]quinoxalin-4-yl)ethane-1,2-diamine

BMS-345541; BMS345541; BMS-345541 free base; BMS345541; N1-(1,8-dimethylimidazo[1,2-a]quinoxalin-4-yl)ethane-1,2-diamine; IKK Inhibitor III, BMS-345541; IKK Inhibitor III; 1,2-Ethanediamine, N-(1,8-dimethylimidazo(1,2-a)quinoxalin-4-yl)-; BMS 345541; UNII-26SU0NEF5F; BMS-345541 free base

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 1 mg/mL (3.92 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 1 mg/mL (3.92 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 1 mg/mL (3.92 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 30% propylene glycol, 5% Tween 80, 65% D5W: 30mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)