| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
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Purity: ≥98%
| Targets |
Endogenous metabolite
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| ln Vitro |
There are two main enzymes that convert tryptophan (Trp) to kynurenine (Kyn): tryptophan-2,3-dioxygenase (TDO) and indoleamine 2,3-dioxygenase (IDO). Kyn accumulation can promote immunosuppression in certain cancers. In this study, we investigated Trp degradation to Kyn by IDO and TDO in primary human hepatocytes (PHH) and tumoral HepG2 cells. To quantify Trp-degradation and Kyn-accumulation, using reversed-phase high-pressure liquid chromatography, the levels of Trp and Kyn were determined in the culture media of PHH and HepG2 cells. The role of IDO in Trp metabolism was investigated by activating IDO with IFN-γ and inhibiting IDO with 1-methyl-tryptophan (1-DL-MT). The role of TDO was investigated using one of two TDO inhibitors: 680C91 or LM10. Real-time PCR was used to measure TDO and IDO expression. Trp was degraded in both PHH and HepG2 cells, but degradation was higher in PHH cells. However, Kyn accumulation was higher in the supernatants of HepG2 cells. Stimulating IDO with IFN-γ did not significantly affect Trp degradation and Kyn accumulation, even though it strongly upregulated IDO expression. Inhibiting IDO with 1-DL-MT also had no effect on Trp degradation. In contrast, inhibiting TDO with 680C91 or LM10 significantly reduced Trp degradation. The expression of TDO but not of IDO correlated positively with Kyn accumulation in the HepG2 cell culture media. Furthermore, TDO degraded L-Trp but not D-Trp in HepG2 cells. Kyn is the main metabolite of Trp degradation by TDO in HepG2 cells. The accumulation of Kyn in HepG2 cells could be a key mechanism for tumor immune resistance. Two TDO inhibitors, 680C91 and LM10, could be useful in immunotherapy for liver cancers [1].
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| ln Vivo |
Indoleamine 2,3 dioxygenase-1 (IDO1) catalyzes tryptophan-kynurenine metabolism in many inflammatory and cancer diseases. Of note, acute inflammation that occurs immediately after heart injury is essential for neonatal cardiomyocyte proliferation and heart regeneration. However, the IDO1-catalyzed tryptophan metabolism during heart regeneration is largely unexplored. Here, we find that apical neonatal mouse heart resection surgery led to rapid and consistent increases in cardiac IDO1 expression and kynurenine accumulation. Cardiac deletion of Ido1 gene or chemical inhibition of IDO1 impairs heart regeneration. Mechanistically, elevated kynurenine triggers cardiomyocyte proliferation by activating the cytoplasmic aryl hydrocarbon receptor-SRC-YAP/ERK pathway. In addition, cardiomyocyte-derived kynurenine transports to endothelial cells and stimulates cardiac angiogenesis by promoting aryl hydrocarbon receptor nuclear translocation and enhancing vascular endothelial growth factor A expression. Notably, Ahr deletion prevents indoleamine 2,3 dioxygenase -kynurenine-associated heart regeneration. In summary, increasing indoleamine 2,3 dioxygenase-derived kynurenine level promotes cardiac regeneration by functioning as an endogenous regulator of cardiomyocyte proliferation and cardiac angiogenesis [2].
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| Cell Assay |
HepG2 cells and PHH cells were cultured as previously described. PHH and HepG2 cells were cultured in William’s medium with increasing concentrations of Trp (60, 120, and 240 µmol/L). Trp and Kyn levels in cultivated PHH and HepG2 cell supernatants were examined at 24 h, 48 h, and 72 h. In the next step, we also investigated the maximum Trp degradation in HepG2 cells by culturing these cells for 72 h in William’s medium containing higher amounts of Trp (480 µmol/L, 960 µmol/L, and 1920 µmol/L). To determine whether IDO is responsible for Trp breakdown, we activated IDO with IFN-γ and inhibited IDO with 1-methyl-tryptophan (1-DL-MT) in PHH and HepG2 cells. In these experiments, the cells were cultured for 72 h under one of the following conditions: no treatment, stimulated with IFN-γ, treated with IFN-γ + 1-DL-MT, and treated with 1-DL-MT alone. After 72 h, the amounts of Trp and Kyn in the supernatants were determined. To determine whether TDO is responsible for Trp breakdown, we cultured cells with the TDO inhibitors LM10 and 680C91. In these experiments, HepG2 cells were treated with increasing amounts of 680C91 (10, 20, 40, and 80 µmol/L) and LM10 (25, 50, and 75 µmol/L) in a medium containing 240 µM Trp for 72 h before measuring Trp and Kyn levels in the supernatant. To investigate which Trp isoform (L-Trp or D-Trp) is degraded by TDO, we cultured HepG2 cells in media containing 60, 120, or 240 µM of L-Trp or D-Trp for 72 h, and measured the Trp and Kyn levels after 72 h [1].
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| Animal Protocol |
Mice [2]
All procedures involving animals were approved by the Institutional Animal Care and Use Committee at Georgia State University (GSU). Cardiomyocyte (CM)-specific knockout Ido1 (Ido1 mKO) mice were developed by crossing floxed Ido1 (Ido1 F/F) mice with Troponin T (Tnt)-Cre mice. In addition, deficiency of Ido1 in endothelial cell (EC) or vascular smooth muscle cell (VSMC) was obtained by crossbreeding Ido1 F/F with Cdh5-Cre or Myh11-Cre and generated Cdh5-Cre; Ido1 F/F (Ido1 ecKO) or Myh11-Cre; Ido1 F/F (Ido1 vsmcKO) transgenic mice. Their respective littermate of Ido1 F/F were used as controls. The transgenic mice and wild-type mice were in a C57BL/6 background and purchased from Jackson Laboratory (Bar Harbor, ME, USA). Tail biopsy DNA was extracted for genotyping according to the Jackson protocol. All mice were housed in temperature-controlled cages under a 12-h light–dark cycle and given free access to water and food at Georgia State University. Neonatal cardiac apex resection surgery [2] The neonatal mouse heart regeneration model was generated by heart apical resection (AR) surgery for 3–4 weeks as described9. Briefly, postnatal day 1 (P1) or P6 mice were subjected to anesthesia by freezing for 3–5 min. A curved forcep was extended into the chest to pull the heart out. Then, the left ventricular apex was truncated with microsurgical scissors until the ventricular chamber was exposed. The mouse chest was sewn up with 8-0 sutures, and the mice were warmed until recovery. The entire procedure required approximately about 10 min. Sham procedures excluded apex amputation. To detect the role of 1-methyl-D-tryptophan (1MT) and kynurenine (Kyn) in heart regeneration, 1MT (100 mg/kg) and Kyn (100 mg/kg) were intraperitoneally injected into wild-type mice every other day from P1 and were subjected to AR surgery at P2 and P6, respectively. The equal volume of PBS was used as vehicle control. |
| References |
[1]. J Clin Med. 2022 Aug 16;11(16):4794.
[2]. Nat Commun. 2022 Oct 26;13(1):6371. |
| Additional Infomation |
Kynurenine is a ketone that is alanine in which one of the methyl hydrogens is substituted by a 2-aminobenzoyl group. It has a role as a human metabolite. It is a substituted aniline, an aromatic ketone and a non-proteinogenic alpha-amino acid. It is a conjugate acid of a kynureninate.
Kynurenine has been reported in Drosophila melanogaster, Psychotria punctata, and other organisms with data available. Kynurenine is a ketone and an amino acid derivative that is synthesized by either tryptophan 2,3-dioxygenase (TDO)- or indoleamine 2,3-dioxygenase (IDO)-mediated oxidation of tryptophan with diverse biological functions, including vasodilatory, immunoregulatory and neuromodulatory activities. Kynurenine is a precursor for niacin. Additionally, kynurenine can be further metabolized into anthranilic acid, kynurenic acid, and 3-hydroxykynurenine; aberrant production of kynurenine is associated with neurological disease-related cognitive deficits and depressive symptoms. Overexpressed in certain cancer cell types, kynurenine could potentially be used as a biomarker to assess cancer risk. A metabolite of the essential amino acid tryptophan metabolized via the tryptophan-kynurenine pathway. |
| Molecular Formula |
C10H12N2O3
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|---|---|
| Molecular Weight |
208.2139
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| Exact Mass |
208.084
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| Elemental Analysis |
C, 57.69; H, 5.81; N, 13.45; O, 23.05
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| CAS # |
343-65-7
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| PubChem CID |
846
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| Appearance |
Light yellow to yellow solid
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
466.6±45.0 °C at 760 mmHg
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| Melting Point |
~235 °C (dec.)
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| Flash Point |
236.0±28.7 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.626
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| LogP |
1.09
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
15
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| Complexity |
255
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(C1=C([H])C([H])=C([H])C([H])=C1N([H])[H])C([H])([H])C([H])(C(=O)O[H])N([H])[H]
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| InChi Key |
YGPSJZOEDVAXAB-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H12N2O3/c11-7-4-2-1-3-6(7)9(13)5-8(12)10(14)15/h1-4,8H,5,11-12H2,(H,14,15)
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| Chemical Name |
alpha-2-Diamino-gamma-oxobenzenebutyric acid
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| Synonyms |
DL-Kynurenine; 2-Amino-4-(2-aminophenyl)-4-oxobutanoic acid
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| HS Tariff Code |
2934.99.03.00
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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) |
1M HCl : 20 mg/mL (~96.05 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 | 4.8028 mL | 24.0142 mL | 48.0284 mL | |
| 5 mM | 0.9606 mL | 4.8028 mL | 9.6057 mL | |
| 10 mM | 0.4803 mL | 2.4014 mL | 4.8028 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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