| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
SIRT1 0.205 μM (IC50) SIRT2 11.5 μM (IC50) SIRT3 >100 μM (IC50)
SIRT1 (IC50 = 98 nM in fluorimetric assay; IC50 = 0.6 μM in radiochemical assay; IC50 = 38 nM against recombinant SIRT1 expressed from mammalian cells) [1] Selective over SIRT2 (>100 μM), SIRT3 (>100 μM), class I and II HDACs (no inhibition at 100 μM), and NAD glycohydrolase (NADase) (no inhibition at 100 μM) [1] |
|---|---|
| ln Vitro |
Compound 2 (SIRT1-IN-1) had minimal impact on HDAC (IC50>100 μM) and SIRT3 (IC50>100 μM)[1].
SIRT1-IN-1 inhibited recombinant human SIRT1 in a fluorimetric assay with an IC50 of 98 nM. In a radiochemical assay using an unlabeled 19-amino-acid peptide, the IC50 was 0.6 μM (2-12 fold higher than in fluorimetric assay). [1] The compound inhibited nicotinamide exchange reaction with an IC50 of 1.5 μM, suggesting it occupies the nicotinamide binding site. [1] In cells, SIRT1-IN-1 penetrated cells and inhibited the deacetylation of p53 at a concentration of 1 μM. [1] It showed no inhibition of a panel of cytochrome P450 enzymes (3A4, 2D6, 1A2, 2C9, 2C19) at 1 μM. [1] It did not affect hERG channel activity when tested at 10 μM. [1] It was highly stable in rat liver microsomes. [1] |
| ln Vivo |
Pharmacokinetic analysis in mice following oral dosing of SIRT1-IN-1 (10 mg/kg) showed quantitative (100%) oral bioavailability and a serum half-life of 136 minutes. [1]
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| Enzyme Assay |
SIRT1 fluorimetric assay: Recombinant human SIRT1 was incubated with Fluor de Lys-SIRT1 peptide substrate (p53-K382-Ac) and NAD in assay buffer. After 45 min at 37°C, Fluor de Lys Developer II was added and incubated for 15 min at 37°C. Fluorescence was measured at excitation 360 nm and emission 460 nm. IC50 values were calculated using XLFit or GraphPad Prism. [1]
Radiochemical assay (nicotinamide release assay): SIRT1 activity was measured using a p53 peptide substrate (residues 368-386 acetylated on lysine 382) and 14C-labeled NAD. The release of 14C nicotinamide was measured using a boronic acid resin capture method. [1] Nicotinamide exchange assay: SIRT1 was incubated with unlabeled nicotinamide (52 μM) to promote exchange; release of 14C nicotinamide from labeled NAD was measured. [1] NAD glycohydrolase (NADase) assay: Purified NADase from pig brain was incubated with NAD (18.55 μM) and compound, and nicotinamide release was measured. [1] Class I/II HDAC assay: HeLa cell extract containing class I/II HDACs was incubated with H4-K16(Ac) substrate and compound, then developed with Developer II. [1] |
| Cell Assay |
Cell permeability and p53 deacetylation: Cells (mammalian) were treated with SIRT1-IN-1 at 1 μM, and the acetylation status of p53 was assessed (inhibition of deacetylation observed). Detailed protocol not provided in this paper; referenced elsewhere (Solomon et al., Mol. Cell. Biol. 2006). [1]
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| Animal Protocol |
Male C57bl/6J mice were dosed intravenously (iv) or by oral gavage with SIRT1-IN-1 at 10 mg/kg in phosphate-buffered saline containing 4% DMSO and 10% cyclodextrin. Plasma was collected at 5, 15, 30, 60, 90 min and 2, 4, 6, 8, 24 h after dosing. Samples were analyzed by LC-MS after solid-phase extraction. [1]
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| ADME/Pharmacokinetics |
Oral bioavailability in mice: 100% (quantitative) [1]
Serum half-life in mice: 136 min after iv dosing [1] Metabolic stability: Highly stable in rat liver microsomes (incubated at 10 μM with rat hepatic microsomes at 37°C, quantified by HPLC/MS at 0,5,15,30,60 min) [1] Cytochrome P450 inhibition: No inhibition of CYP3A4, 2D6, 1A2, 2C9, 2C19 at 1 μM [1] hERG channel: No effect at 10 μM [1] |
| References | |
| Additional Infomation |
Mechanism of action: Mixed-type inhibitor against both NAD and acetyl-peptide substrates; pattern approaches uncompetitive inhibition, especially against NAD, suggesting binding occurs after binding of both substrates and after release of nicotinamide. The inhibitor likely prevents release of deacetylated peptide and O-acetyl-ADP-ribose. [1]
Structure-activity relationship: Primary carboxamide at the 1-position is essential; removal or modification leads to loss of activity. The 6-chloro substitution is preferred. The (S)-enantiomer (e1) is active (IC50=0.123 μM) while the (R)-enantiomer (e2) is inactive (>100 μM). [1] Potential therapeutic uses: Chemical tool to study SIRT1 biology and explore therapeutic applications for SIRT1 inhibitors. [1] |
| Molecular Formula |
C14H16N2O
|
|---|---|
| Molecular Weight |
228.29
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| Exact Mass |
228.126
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| CAS # |
352554-02-0
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| PubChem CID |
661302
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
2.2
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
1
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
17
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| Complexity |
317
|
| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=C2C(C3=C(N2)C(C(N)=O)CCC3)=C1
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| InChi Key |
KQWPYXBWZWLRMZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H16N2O/c1-8-5-6-12-11(7-8)9-3-2-4-10(14(15)17)13(9)16-12/h5-7,10,16H,2-4H2,1H3,(H2,15,17)
|
| Chemical Name |
6-methyl-2,3,4,9-tetrahydro-1H-carbazole-1-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 : 100 mg/mL (438.04 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (21.90 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 50.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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. Solubility in Formulation 2: ≥ 5 mg/mL (21.90 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 50.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.3804 mL | 21.9020 mL | 43.8039 mL | |
| 5 mM | 0.8761 mL | 4.3804 mL | 8.7608 mL | |
| 10 mM | 0.4380 mL | 2.1902 mL | 4.3804 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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