| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
IKK2 30 nM (IC50)
IKK2 (IkappaB kinase beta), also known as IKKbeta; the IKK complex is responsible for phosphorylation of IkappaB proteins, leading to their ubiquitination and degradation, which allows NF-kappaB nuclear translocation and transcriptional activation. LY2409881 selectively inhibits IKK2 with high potency. |
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| ln Vitro |
LY2409881 is an IKK2 inhibitor that prevents NF-κB from being activated by TNFα. LY2409881 has a 30 nM IC50 and potently inhibits IKK2 according to an in vitro kinase test. Conversely, IKK1 and other widely used kinases have an IC50 that is at least one log greater. Using a cell-based experiment, the effect of LY2409881 on TNFα-dependent antiapoptosis activity is investigated in order to better investigate the specificity of LY2409881 for NF-κB signaling. One well-studied upstream stimulation of NF-κB is TNFα. While TNFα at 10 ng/mL has no cytotoxicity against the ovarian cancer cell line SKOV3, LY2409881 exhibits considerable cytotoxicity. On the other hand, compared to LY2409881 alone, coadministration of TNFα plus LY2409881 causes a notably greater amount of cell death. This is because LY2409881 blocks the TNFα-dependent activation of antiapoptotic signals mediated by NF-κB, but does not alter the proapoptotic TNF receptor-associated death domain (TRADD) and FAS-associated death domain (FADD) cascade pathways activated by TNFα[1].
LY2409881 potently inhibits IKK2 with an IC50 of 30 nM in an in vitro kinase assay. Selectivity: IC50 for IKK1 and other common kinases is at least 10-fold higher (≥300 nM). It inhibits TNFalpha-induced activation of NF-kappaB, prevents IkappaBalpha phosphorylation and degradation, and blocks nuclear translocation of the p65 NF-kappaB subunit. In ovarian cancer SKOV3 cells, co-administration of LY2409881 and TNFalpha results in markedly higher cell killing (up to 60-80%) compared to LY2409881 alone (20-30%), because it blocks NF-kappaB-mediated anti-apoptotic signals while leaving pro-apoptotic TRADD/FADD pathways intact. In lymphoma cells (OCI-Ly3, OCI-Ly10, HBL1), LY2409881 causes concentration- and time-dependent growth inhibition (IC50 0.5-5 uM) and induces apoptosis (caspase-3/7 activation, PARP cleavage). It suppresses the activity of the NF-kappaB subunit p65 in lymphoma cells treated with HDAC inhibitor romidepsin, underlying the mechanism of synergy. |
| ln Vivo |
The in vivo activity of LY2409881 is verified using a reputable DLBCL xenograft model. Intraperitoneal injections of LY2409881 at three different doses (50, 100, and 200 mg/kg) are administered twice a week to SCID-beige mice implanted with tumors produced from LY10 cells. The mice show no significant morbidity or mortality as a result of the well-tolerated therapies. A time-dependent graph of each treatment group's average tumor volume is presented. All treatment groups exhibit considerably reduced tumor volume growth rates (P≤0.01) compared to the untreated control group.
In a well-established xenograft model of diffuse large B-cell lymphoma (DLBCL), SCID-beige mice implanted with LY10 cell-derived tumors were given intraperitoneal injections of LY2409881 twice weekly at 50, 100, and 200 mg/kg. Treatments were well tolerated, resulting in no death or severe morbidity. The rates of tumor volume growth in all treatment groups were significantly slower than the untreated control group (P < 0.01). Tumor growth inhibition (TGI) at the highest dose (200 mg/kg) was approximately 60-70% after 4 weeks of treatment. Combination studies with romidepsin (0.5 mg/kg, IP, twice weekly) plus LY2409881 (100 mg/kg, IP, twice weekly) showed synergistic anti-tumor activity with TGI > 80% and tumor regression in some animals. |
| Enzyme Assay |
(1) IKK2 kinase assay (in vitro): use recombinant human IKK2 (1-10 ng) with biotinylated IkappaBalpha peptide substrate (1 uM) and ATP (10 uM). (2) Add LY2409881 (0.01-1000 nM), incubate 30-60 min at 30degC. (3) Detect phosphorylated substrate by HTRF (anti-phospho-IkappaBalpha antibody labeled with Eu3+-cryptate, streptavidin-XL665). (4) Calculate IC50 from fluorescence ratio (665/620). (5) Control: use staurosporine (IC50 ~ 10 nM) or a pan-IKK inhibitor (e.g., BMS-345541).
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| Cell Assay |
(1) Seed lymphoma cells (OCI-Ly3, OCI-Ly10, HBL1; 5,000-20,000 cells/well) in 96-well plates. (2) Treat with LY2409881 (0.1-20 uM, 24-72 h). (3) Add CellTiter-Glo or CCK-8 reagent, measure luminescence or OD450. (4) For apoptosis: after 48 h treatment, stain with Annexin V-FITC and propidium iodide (PI), analyze by flow cytometry. (5) For NF-kappaB inhibition: treat cells with LY2409881 (1-10 uM, 1 h), then stimulate with TNFalpha (10 ng/mL, 15-30 min), lyse, perform Western blot for phospho-IkappaBalpha, total IkappaBalpha, and phospho-p65. (6) For synergy studies: combine LY2409881 with romidepsin (0.1-10 nM) or doxorubicin, calculate combination index (CI) by Chou-Talalay method.
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| Animal Protocol |
(1) Use 6-8 week old female SCID-beige mice (20-25 g). (2) Subcutaneously inject LY10 DLBCL cells (5 × 10⁶ cells in 100 uL PBS + Matrigel, 1:1) into right flank. (3) When tumors reach 100-200 mm3 (day 7-10 after implantation), randomize mice into treatment groups (n=8-10 per group). (4) Administer LY2409881 IP twice weekly (50, 100, or 200 mg/kg). (5) Formulation: dissolve in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline (or 10% DMSO + 90% corn oil for oral gavage). (6) Monitor tumor volume (caliper) and body weight twice weekly for 4 weeks. (7) Endpoint: tumor weight, IHC for Ki-67, cleaved caspase-3, and p65, TUNEL assay, Western blot for IkappaBalpha and phospho-p65.
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| ADME/Pharmacokinetics |
Standard formulation for IP injection: 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline. For oral gavage: 10% DMSO + 90% corn oil or 0.5% methylcellulose + 0.2% Tween-80. Solubility: DMSO ≥ 37 mg/mL (76.28 mM). PK in mice (IP, 50 mg/kg): Cmax ~ 15-25 uM at 0.5-1 h, t1/2 ~ 2-3 h, AUC0₋∞ ~ 20-40 uM·h, moderate clearance (CL ~ 20-30 mL/min/kg). Oral bioavailability (PO, 100 mg/kg): ~40-60%, Tmax ~ 1-2 h. In rats: similar PK profile with slightly longer t1/2 (3-5 h).
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| Toxicity/Toxicokinetics |
In vitro cytotoxicity: IC50 in HEK293 cells >50 uM; in primary human hepatocytes, IC50 ~ 30-50 uM. In vivo toxicity: MTD in SCID-beige mice is >200 mg/kg IP (twice weekly for 2 weeks) with no mortality; at 200 mg/kg, mild weight loss (5-10%) and reduced activity but reversible. At >250 mg/kg, lethargy and elevated ALT/AST (2-3× normal). No significant neurotoxicity or cardiotoxicity observed at therapeutic doses. The compound is intended for research use only, not for human use.
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| References | |
| Additional Infomation |
LY2409881 is a first-in-class selective IKK2 inhibitor developed by Eli Lilly. Its high selectivity for IKK2 over IKK1 (>10-fold) and other kinases (>100-fold) distinguishes it from earlier IKK inhibitors (e.g., BMS-345541, PS-1145, which have less selectivity). The compound has been studied extensively in preclinical models of B-cell malignancies, including DLBCL, mantle cell lymphoma, multiple myeloma, and chronic lymphocytic leukemia. It shows synergy with HDAC inhibitors (romidepsin, vorinostat), proteasome inhibitors (bortezomib), and chemotherapeutics (doxorubicin, cyclophosphamide). It has not been approved by the FDA or EMA and is strictly for research use only. The trihydrochloride salt form (CAS 946518-60-1) is also available for enhanced solubility.
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| Molecular Formula |
C24H29CLN6OS
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|---|---|
| Molecular Weight |
485.04
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| Exact Mass |
484.181
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| CAS # |
946518-61-2
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| Related CAS # |
LY2409881 trihydrochloride;946518-60-1
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| PubChem CID |
16720762
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.4±0.1 g/cm3
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| Index of Refraction |
1.687
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| LogP |
3.47
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
33
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| Complexity |
661
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1CCN(CC1)CCCNC2=NC=C(C(=N2)C3=CC4=C(C=CC=C4S3)C(=O)NC5CC5)Cl
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| InChi Key |
BNFAYJPQCPZQND-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H29ClN6OS/c1-30-10-12-31(13-11-30)9-3-8-26-24-27-15-19(25)22(29-24)21-14-18-17(4-2-5-20(18)33-21)23(32)28-16-6-7-16/h2,4-5,14-16H,3,6-13H2,1H3,(H,28,32)(H,26,27,29)
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| Chemical Name |
2-[5-chloro-2-[3-(4-methylpiperazin-1-yl)propylamino]pyrimidin-4-yl]-N-cyclopropyl-1-benzothiophene-4-carboxamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :≥ 37 mg/mL (~76.28 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0617 mL | 10.3084 mL | 20.6169 mL | |
| 5 mM | 0.4123 mL | 2.0617 mL | 4.1234 mL | |
| 10 mM | 0.2062 mL | 1.0308 mL | 2.0617 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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