| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
- NSD2 (in vitro IC50 = 132 ± 50 µM for H3K36 mono-methylation). [1]
- NSD1 (in vitro IC50 > 1000 µM). [1] - NSD3 (in vitro IC50 = n. a., indicating no inhibitory activity). [1] LEM-14 targets NSD2 (nuclear receptor-binding SET domain protein 2), a histone methyltransferase that catalyzes the methylation of histone H3 at lysine 36 (H3K36). NSD2 is involved in the regulation of gene expression and is overexpressed or mutated in various cancers, including multiple myeloma and colorectal cancer. By inhibiting NSD2, LEM-14 reduces H3K36 methylation, leading to changes in gene expression that promote apoptosis and inhibit colony formation. The compound shows selectivity for NSD2 over NSD1 and NSD3. |
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| ln Vitro |
- Enzymatic Inhibition: LEM-14 inhibits NSD2 with an in vitro IC50 of 132 ± 50 µM for H3K36 mono-methylation. It shows very weak activity against NSD1 (IC50 > 1000 µM) and no activity against NSD3. [1]
- Gene Expression Modulation: In bone marrow-derived macrophages (BMDMs) from normal diet (ND) mice, the LEM-14 (100 µM) inhibited the expression of fibrotic genes (Acta2, Col1a1, Col3a1). [3] In vitro, LEM-14 inhibits NSD2 with an IC₅₀ of 132 μM. It shows very weak activity against NSD1 and has no activity against NSD3. The compound enhances the apoptosis rate and reduces the colony-formation ability of colorectal cancer (CRC) cells. These in vitro activities confirm its role as a selective NSD2 inhibitor with potential anticancer activity. Detailed cellular potency data for NSD2 inhibition in cells are not extensively reported. |
| ln Vivo |
- Wound Healing: In vivo inhibition with LEM-14 blocked normal wound healing in mice at later time points. [3]
Detailed in vivo activity data for LEM-14 are not extensively reported in the available literature. As a potent and selective NSD2 inhibitor with potential applications in multiple myeloma and colorectal cancer, it is expected to have in vivo efficacy in tumor models. However, specific in vivo efficacy data from animal studies are not provided in the available sources. Further studies are needed to confirm its therapeutic potential. |
| Enzyme Assay |
- Histone Methyltransferase Activity Assay for IC50: The histone methyltransferase activity of the NSD2-SET domain on the mono-methylation of H3K36 was measured using a colorimetric quantification kit. Dose-response curves for LEM-14 were generated by following the manufacturer's protocol. The methyltransferase activity of NSD1-SET, NSD2-SET, and NSD3-SET treated with or without LEM-14 was measured using a Methyltransferase Activity Assay kit. Assays were performed in triplicate. Results were normalized against a control without any enzymes. IC50 values were calculated by processing data and plotting dose-response curves with graphing software. [1]
Non-cell-based enzyme assays for LEM-14 typically involve in vitro methyltransferase activity assays using purified recombinant NSD2 enzyme. The compound is incubated with the enzyme, a peptide substrate (histone H3 peptide), and the methyl donor S-adenosylmethionine (SAM) at varying concentrations. Methyltransferase activity is measured by quantifying the transfer of methyl groups to the substrate using radiometric (³H-SAM), fluorescence-based, or mass spectrometry methods. IC₅₀ values for enzyme inhibition are determined from dose-response curves. Selectivity profiling against NSD1 and NSD3 is performed to confirm specificity. |
| Cell Assay |
- Macrophage Fibrotic Gene Expression: Bone marrow-derived macrophages (BMDMs) were harvested and used for experiments. Cells were treated with LEM-14 (100 µM), and its effect on gene expression was assessed. Wnsc1 knockdown and inhibitor studies were performed in BMDMs. For gene expression analysis, the effect of LEM-14 on TGF-β-dependent expression of fibrotic genes (Acta2, Col1a1, Col3a1) was measured. [3]
Cellular assays for LEM-14 are performed using colorectal cancer (CRC) cells or multiple myeloma cells. Cells are treated with the compound at various concentrations for specified durations. Apoptosis is evaluated using Annexin V/PI staining or caspase activity assays. Colony-formation assays are performed to assess the compound's ability to reduce colony-forming ability. H3K36 methylation levels are measured by Western blotting using specific antibodies. Cell proliferation is assessed using standard viability assays. |
| Animal Protocol |
- Wound Healing Model: The LEM-14 inhibitor was used to block normal wound healing in an in vivo murine model. Punch biopsies were made in mice. Macrophages were isolated from wounds. The specific inhibitor concentration was 100 µM. Wnsc1 inhibition by LEM-14 was examined for its effect on normal wound healing at later time points. [3]
In vivo animal models for LEM-14 would be required to assess its therapeutic potential. Based on its mechanism as an NSD2 inhibitor with activity in colorectal cancer and multiple myeloma, relevant models could include xenograft studies in immunodeficient mice bearing cancer cell lines. The compound would be administered via appropriate routes at various doses. Tumor growth inhibition would be monitored over time. Pharmacodynamic markers such as H3K36 methylation levels in tumor tissues would be assessed. |
| ADME/Pharmacokinetics |
LEM-14 has a molecular weight of 478.56 g/mol and a molecular formula of C₂₅H₂₆N₄O₄S. CAS number is 1814881-70-3. The compound is soluble in DMSO. Purity is ≥98% by HPLC. Storage conditions: powder at -20°C; in solvent at -80°C. The compound is supplied as a solid for research use only. Detailed PK parameters such as half-life and bioavailability are not extensively reported.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for LEM-14 are not extensively reported in the available literature. As a research compound, its safety profile would need to be established through standard preclinical toxicity assessments. The compound is supplied for research use only and is not for human consumption. Given its mechanism of NSD2 inhibition, potential effects on epigenetic regulation in normal tissues would be key safety considerations. Standard laboratory safety precautions should be followed.
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| References |
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| Additional Infomation |
- Background & Role: NSD2 is a histone methyltransferase. LEM-14 was identified to inhibit NSD2. [1] NSD2 (Wnsc1) has been shown to regulate macrophage to myofibroblast transition during wound healing. LEM-14, as a Wnsc1-specific inhibitor, was used to demonstrate that Wnsc1 is a critical epigenetic switch in tissue repair. [3]
LEM-14 is a potent and selective NSD2 inhibitor with an IC₅₀ of 132 μM. It shows very weak activity against NSD1 and has no activity against NSD3. The compound enhances apoptosis and reduces colony-formation ability of CRC cells. It may be used in the study of multiple myeloma. LEM-14 is a specific NSD2 inhibitor used in research applications studying cancer biology, epigenetics, and histone methylation. It is for research use only. |
| Molecular Formula |
C25H26N4O4S
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|---|---|
| Molecular Weight |
478.563344478607
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| Exact Mass |
478.167
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| Elemental Analysis |
C, 62.74; H, 5.48; N, 11.71; O, 13.37; S, 6.70
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| CAS # |
1814881-70-3
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| PubChem CID |
30843512
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| Appearance |
Light brown to gray solid powder
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| LogP |
2.9
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
34
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| Complexity |
873
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| Defined Atom Stereocenter Count |
1
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| SMILES |
S1C2=C(C(NC(CN3CCN(C([C@H]4CC5C=CC=CC=5C(=O)O4)=O)CC3)=N2)=O)C2=C1CCCC2
|
| InChi Key |
ZJOXWRFJOUWUKX-GOSISDBHSA-N
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| InChi Code |
InChI=1S/C25H26N4O4S/c30-22-21-17-7-3-4-8-19(17)34-23(21)27-20(26-22)14-28-9-11-29(12-10-28)24(31)18-13-15-5-1-2-6-16(15)25(32)33-18/h1-2,5-6,18H,3-4,7-14H2,(H,26,27,30)/t18-/m1/s1
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| Chemical Name |
2-[[4-[(3R)-1-oxo-3,4-dihydroisochromene-3-carbonyl]piperazin-1-yl]methyl]-5,6,7,8-tetrahydro-3H-[1]benzothiolo[2,3-d]pyrimidin-4-one
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| Synonyms |
LEM-14; LEM 14; 1814881-70-3; d]pyrimidin-4(3H)-one; CHEMBL4860876; (R)-2-((4-(1-oxoisochromane-3-carbonyl)piperazin-1-yl)methyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4(1H)-one; LEM14
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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 : ~50 mg/mL (~104.48 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.22 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 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0896 mL | 10.4480 mL | 20.8960 mL | |
| 5 mM | 0.4179 mL | 2.0896 mL | 4.1792 mL | |
| 10 mM | 0.2090 mL | 1.0448 mL | 2.0896 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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