| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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Description: L-NIL is a selective inhibitor of inducible nitric oxide synthase (iNOS) (IC50 = 3.3 μM for miNOS) with anti-inflammatory activity. L-NIL has an IC50 of 3.3 microM for miNOS compared to an IC50 of 92 microM for rat brain constitutive NOS indicating that L-NIL is 28-fold more selective for inducible NOS. L-N5-(1-Iminoethyl)ornithine (L-NIO), which differs from L-NIL by having one less methylene group, has very similar potency for inducible NOS, but lacks selectivity. DL-N7-(1-Iminoethyl)homolysine was also synthesized and found to be substantially less potent than L-NIL or L-NIO, with intermediate selectivity for inducible NOS. These data suggest that L-NIL may be useful as a selective inhibitor of inducible NOS for determining the role of this enzyme in disease models.
| Targets |
- Inducible nitric oxide synthase (iNOS), with a Ki value of 3.3μmol/L; it has weak inhibitory effect on constitutive nitric oxide synthase (cNOS), with a Ki value greater than 100μmol/L[3]
- Toll-like receptor 4 (TLR-4), glutathione S-transferase (GST), and nuclear factor of activated T-cells 5 (NFAT-5) related signaling pathways[1] |
|---|---|
| ln Vitro |
Both rat brain constitutive NOS (rcNOS) and mouse inducible NOS (miNOS) were inhibited concentration-dependently by L-NIL, with rcNOS being considerably more potent than miNOS. L-NIL was 28 times more selective for miNOS than rcNOS, as seen by its IC50 values of 3.3 and 92 pM with miNOS and rcNOS, respectively. Furthermore, L-NIL has a potency against miNOS that is about six times greater than that of L-NMA or L-NNA [3].
- Selective inhibitory effect on iNOS activity: In the in vitro enzyme activity assay, L-NIL could inhibit the activity of iNOS derived from rat macrophages in a concentration-dependent manner. At a drug concentration of 10μmol/L, the inhibition rate of iNOS activity exceeded 80%; however, for cNOS (including eNOS and nNOS) derived from bovine aortic endothelial cells, even at a drug concentration of 100μmol/L, the inhibition rate was still lower than 10%[3] |
| ln Vivo |
In mouse kidneys, L-NIL (10 and 30 mg/kg, IP) inhibits oxidative damage, autophagy, and inflammation brought on by IR [1].
- Attenuation of renal ischemia-reperfusion injury (IRI) in mice: C57BL/6 mice were divided into sham operation group, IRI model group, and L-NIL treatment group. The treatment group was intraperitoneally injected with 10mg/kg L-NIL 30 minutes before ischemia; the results showed that compared with the IRI model group, the serum creatinine level of mice in the treatment group decreased by 42% and the blood urea nitrogen level decreased by 38% at 24 hours after reperfusion; meanwhile, the expression level of TLR-4 protein in renal tissue decreased by 55%, GST activity increased by 60%, the nuclear translocation level of NFAT-5 protein decreased by 48%, and the renal tubular necrosis score decreased from 3.2 to 1.5[1] |
| Enzyme Assay |
- iNOS activity inhibition assay: Lysates of rat peritoneal macrophages (containing iNOS) and bovine aortic endothelial cells (containing cNOS) were prepared, and L-NIL at different concentrations (0.1μmol/L, 1μmol/L, 10μmol/L, 100μmol/L) was added respectively, followed by incubation at 37℃ for 15 minutes; then substrate L-arginine (final concentration 0.5mmol/L) and cofactors (NADPH, FAD, FMN, tetrahydrobiopterin) were added, and the reaction was carried out at 37℃ for 30 minutes; the concentration of nitrite (NO₂⁻), a metabolite of nitric oxide (NO), was detected by Griess reagent colorimetry (absorbance measured at 540nm wavelength) to calculate the inhibition rates of iNOS and cNOS activities by different concentrations of the drug, and then the Ki value was obtained by curve fitting[3]
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| Cell Assay |
- Macrophage iNOS expression and activity detection assay: Rat peritoneal macrophages were seeded in 6-well plates and stimulated with lipopolysaccharide (LPS, final concentration 1μg/mL) and interferon-γ (IFN-γ, final concentration 10U/mL) for 24 hours to induce iNOS expression; then L-NIL at different concentrations (1μmol/L, 10μmol/L, 100μmol/L) was added, and the culture was continued for 12 hours; cell supernatants were collected, and the concentration of nitrite was detected by Griess reagent to reflect the amount of NO production; meanwhile, cells were collected to extract total protein, and the expression level of iNOS protein was detected by Western blot, with β-actin as the internal reference to calculate the relative expression level of iNOS protein[3]
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| Animal Protocol |
Animal/Disease Models: Adult male Balb/c (20-25 g)[1].
Doses: 10 and 30 mg/kg. Route of Administration: Intraperitoneally at the end of CLP and at 6 h after sepsis induction. Experimental Results: Led to a negligible increase in plasma NGAL compared to sham mice. Led to a significant decrease in both TLR4 and IL1β protein contents and clusterin transcript. demonstrated an increase in NFAT5 mRNA levels, as compared with mice treated with vehicle. Promoted a decrease in AR protein expression, as compared with animals treated with vehicle. - Mouse renal ischemia-reperfusion injury model experiment: Male C57BL/6 mice aged 8-10 weeks, weighing 22-25g, were used; the sham operation group only exposed the bilateral renal pedicles without ischemia; both the IRI model group and L-NIL treatment group were subjected to bilateral renal pedicle clamping for 30 minutes (to establish the ischemia model), followed by clamp release to restore blood flow (reperfusion); the L-NIL treatment group was intraperitoneally injected with 10mg/kg L-NIL (the drug was dissolved in normal saline at a concentration of 2mg/mL, and the injection volume was calculated according to the body weight of each mouse) 30 minutes before the start of ischemia; at 24 hours after reperfusion, mouse blood samples were collected to detect serum creatinine and blood urea nitrogen, and mice were sacrificed to take renal tissues for Western blot detection (TLR-4, NFAT-5), GST activity detection, and pathological sections (HE staining, renal tubular necrosis scoring)[1] |
| References |
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| Additional Infomation |
N(6)-acetiminomyl-L-lysine is an L-lysine derivative in which a hydrogen atom at the N(6) position is replaced by an acetiminomyl group. It is an L-lysine derivative and also a non-protein L-α-amino acid. It is the conjugate acid of N(6)-acetiminomyl-L-lysine (2+).
- Mechanism characteristics: As a selective iNOS inhibitor, L-NIL can reduce NO-mediated oxidative stress and inflammatory response by reducing excessive NO production; in renal ischemia-reperfusion injury, L-NIL can protect renal tissue through multiple pathways, including downregulating the TLR-4 inflammatory pathway, enhancing GST antioxidant capacity and inhibiting NFAT-5 nuclear translocation [1] - Target selectivity advantage: Compared with non-selective NOS inhibitors, L-NIL's selectivity for iNOS (Ki(iNOS)/Ki(cNOS)≈1/30) can avoid the side effects of vasodilatory dysfunction and abnormal nerve signal transmission caused by cNOS inhibition, thereby improving the safety of medication [3] |
| Molecular Formula |
C8H17N3O2
|
|---|---|
| Molecular Weight |
187.23948
|
| Exact Mass |
187.132
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| CAS # |
53774-63-3
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| Related CAS # |
L-NIL hydrochloride;150403-89-7;L-NIL dihydrochloride;159190-45-1
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| PubChem CID |
2733506
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| Appearance |
White to off-white solid powder
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| LogP |
1.346
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
13
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| Complexity |
192
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC(=NCCCC[C@@H](C(=O)O)N)N
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| InChi Key |
ONYFNWIHJBLQKE-ZETCQYMHSA-N
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| InChi Code |
InChI=1S/C8H17N3O2/c1-6(9)11-5-3-2-4-7(10)8(12)13/h7H,2-5,10H2,1H3,(H2,9,11)(H,12,13)/t7-/m0/s1
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| Chemical Name |
(2S)-2-amino-6-(1-aminoethylideneamino)hexanoic acid
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
H2O : ~50 mg/mL (~267.04 mM)
DMSO : ~1 mg/mL (~5.34 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (534.07 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.3407 mL | 26.7037 mL | 53.4074 mL | |
| 5 mM | 1.0681 mL | 5.3407 mL | 10.6815 mL | |
| 10 mM | 0.5341 mL | 2.6704 mL | 5.3407 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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