| Size | Price | Stock | Qty |
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| 100mg |
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| Other Sizes |
| Targets |
5-Methoxy-DL-tryptophan (5-MTP) primarily targets endothelial cells (ECs), vascular smooth muscle cells (SMCs), and macrophages, functioning as an endothelial factor with potent anti-inflammatory properties. Its primary mechanism of action involves the inhibition of two key inflammatory signaling cascades: the p38 mitogen-activated protein kinase (MAPK) pathway and the nuclear factor kappa-B (NF-κB) pathway. By blocking the activation of these pathways, 5-MTP effectively suppresses the expression of multiple pro-inflammatory mediators. Specifically, it is a potent inhibitor of cyclooxygenase-2 (COX-2) expression, a key enzyme in the inflammatory cascade that produces prostaglandins. In contrast, other 5-hydroxyindole metabolites of L-tryptophan, such as serotonin, do not exhibit this regulatory effect on COX-2. While melatonin also possesses COX-2 suppressing properties, it requires significantly higher, supra-pharmacological concentrations to achieve a similar effect. Beyond its anti-inflammatory actions, 5-MTP targets fibrotic pathways by inhibiting the differentiation of fibroblasts into myofibroblasts through downregulation of the transforming growth factor-beta (TGF-β)/SMAD3 and phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathways. This dual targeting of inflammatory and fibrotic pathways makes 5-MTP a unique modulator of tissue repair and pathological remodeling. The compound also exhibits protective effects on the vascular endothelium and can attenuate intimal hyperplasia following arterial injury, indicating targeting of vascular smooth muscle cell proliferation and migration. The L-enantiomer is believed to be the primary biologically active form, as it is the endogenous metabolite, while the specific activity of the D-isomer remains to be fully elucidated.
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| ln Vitro |
In vitro, 5-Methoxy-DL-tryptophan demonstrates broad-spectrum suppression of pro-inflammatory cytokines and mediators. At a concentration of 50 μM, it significantly reduces lipopolysaccharide (LPS)-induced release of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and cyclooxygenase-2 (COX-2) in RAW 264.7 macrophages. This robust anti-inflammatory activity serves as a benchmark for high-throughput screening calibration. The compound's anti-inflammatory efficacy has been compared to other tryptophan metabolites; 5-MTP is a potent COX-2 inhibitor, whereas serotonin does not control COX-2 expression. Melatonin reduces LPS-induced IL-6 levels in vitro but often requires high doses. In addition to its anti-inflammatory effects, 5-MTP has demonstrated potent anti-fibrotic activity in various models of tissue fibrosis, including in the lungs, kidneys, and liver. It inhibits the differentiation of fibroblasts into myofibroblasts, a key process in the development of fibrosis, by downregulating the TGF-β/SMAD3 and PI3K/AKT signaling pathways. The compound also exhibits protective effects on the vascular endothelium and can attenuate intimal hyperplasia following arterial injury. The L-enantiomer is believed to be the primary biologically active form, as it is the endogenous metabolite, while the specific activity of the D-isomer remains an area for further investigation. These in vitro findings establish 5-MTP as a potent and specific anti-inflammatory and anti-fibrotic agent with a distinct mechanism of action compared to other tryptophan metabolites.
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| ln Vivo |
In vivo, 5-Methoxy-DL-tryptophan has been validated in mouse models of systemic inflammation and atherosclerosis. In LPS-induced endotoxemia mouse models, administration of 5-MTP at 23.4 mg/kg via intraperitoneal injection (twice weekly for 12-20 weeks) significantly increases survival rates. The compound attenuates arterial denudation-induced intimal hyperplasia via opposing effects on vascular endothelial and smooth muscle cells. In a mouse model of atherosclerosis, 5-MTP (23.5 mg/kg, i.p., twice weekly for 12-20 weeks) attenuates Oil Red O staining of the aortic arch, indicating reduced atherosclerotic lesion formation. The anti-inflammatory action of 5-MTP in vivo is further supported by its ability to block the activation of p38 MAPK and NF-κB, key signaling pathways in the inflammatory cascade. Studies have shown that 5-MTP provides protection from systemic inflammation and functions as an immunomodulator. While direct comparative data with melatonin in vivo is limited, the existing evidence suggests a more potent and specific anti-inflammatory profile for 5-MTP, particularly in the context of COX-2 inhibition. The compound's efficacy in both acute inflammation models (endotoxemia) and chronic inflammatory conditions (atherosclerosis) highlights its broad therapeutic potential. These in vivo findings are consistent with the compound's in vitro mechanism of action and support its continued development as a potential therapeutic agent for inflammatory and cardiovascular diseases. The validated in vivo efficacy at well-tolerated doses establishes 5-MTP as a promising lead compound for further preclinical and potentially clinical development.
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| Enzyme Assay |
In vitro enzyme and receptor binding assays for 5-Methoxy-DL-tryptophan typically involve the use of isolated enzyme preparations or receptor-containing membrane fractions to study its mechanism of action. For assessing COX-2 inhibition, assays are performed using purified COX-2 enzyme or cell lysates from LPS-stimulated macrophages, with enzyme activity measured by prostaglandin production using ELISA or colorimetric methods. To evaluate the compound's effects on signaling pathways, kinase activity assays are conducted using immunoprecipitated p38 MAPK or NF-κB components, with phosphorylation status determined by western blotting or ELISA. Receptor binding studies for 5-MTP are performed using radiolabeled or fluorescently labeled ligands in competition binding experiments with membrane preparations from cells expressing target receptors. The compound's binding affinity to potential targets such as G-protein coupled receptors or nuclear receptors can be determined using scintillation proximity assays or fluorescence polarization techniques. For assessing anti-fibrotic activity, assays are conducted using isolated TGF-β receptors or downstream signaling components, with SMAD3 phosphorylation measured by phospho-specific antibodies. These cell-free systems allow for the precise characterization of 5-MTP's molecular interactions without the confounding effects of cellular metabolism or transport. Typical assay conditions involve incubation at 37°C for 30-60 minutes in appropriate buffer systems (pH 7.4), with reaction termination by addition of stop solution or rapid cooling. Results are expressed as IC₅₀ values for inhibition or EC₅₀ values for activation, with appropriate positive and negative controls included in each experiment. The metabolic stability of the methoxy scaffold ensures reproducible results across different assay platforms.
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| Cell Assay |
In vitro cell-based assays for 5-Methoxy-DL-tryptophan are typically performed using macrophage cell lines such as RAW 264.7, which serve as a robust model for studying inflammatory responses. The standard protocol involves seeding cells in appropriate culture medium (e.g., DMEM supplemented with 10% fetal bovine serum) and allowing them to adhere overnight. Cells are then pretreated with 5-MTP at various concentrations (typically ranging from 1-100 μM) for 1-2 hours, followed by stimulation with lipopolysaccharide (LPS) at 1 μg/mL for 4-24 hours to induce an inflammatory response. After stimulation, culture supernatants are collected for cytokine analysis using enzyme-linked immunosorbent assay (ELISA) or multiplex bead-based assays to measure levels of TNF-α, IL-1β, IL-6, and other inflammatory mediators. Cell lysates are prepared for western blotting to assess the phosphorylation status of p38 MAPK, NF-κB, and other signaling proteins. For anti-fibrotic studies, fibroblasts are cultured and stimulated with TGF-β in the presence or absence of 5-MTP, with myofibroblast differentiation assessed by α-smooth muscle actin expression and collagen production. For vascular protection studies, endothelial cells or vascular smooth muscle cells are used to assess the compound's effects on cell proliferation, migration, and barrier function. Cell viability is routinely monitored using MTT or CCK-8 assays to ensure that observed effects are not due to cytotoxicity. The compound is typically dissolved in DMSO as a stock solution and diluted in culture medium to the desired final concentration, with the final DMSO concentration kept below 0.1% to avoid solvent effects. Each experiment includes appropriate controls (untreated, LPS-only, and vehicle controls) and is performed in triplicate to ensure statistical reliability.
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| Animal Protocol |
In vivo animal experiments for 5-Methoxy-DL-tryptophan are conducted using mouse models of inflammation and vascular disease. For endotoxemia studies, 8-12 week old C57BL/6 mice are administered 5-MTP at 23.4 mg/kg via intraperitoneal injection, followed by LPS challenge (typically 10-20 mg/kg, i.p.) to induce systemic inflammation. Survival is monitored over 24-72 hours, and blood/tissue samples are collected for cytokine analysis and histopathological examination. For atherosclerosis studies, ApoE⁻/⁻ mice fed a high-fat diet are treated with 5-MTP at 23.5 mg/kg via intraperitoneal injection twice weekly for 12-20 weeks. At study termination, the aortic arch and entire aorta are dissected, stained with Oil Red O to visualize atherosclerotic lesions, and lesion area is quantified by image analysis. For arterial injury models, mice undergo femoral artery denudation using a wire injury technique, followed by 5-MTP administration to assess neointimal hyperplasia. Tissue sections are stained with hematoxylin and eosin or elastin van Gieson to measure intima-to-media ratios. For pharmacokinetic studies, blood samples are collected at various time points post-administration, and plasma concentrations of 5-MTP are measured by LC-MS/MS. All animal procedures are conducted in accordance with institutional animal care and use committee guidelines, with appropriate sample sizes (typically n=6-10 per group) to ensure statistical power. The compound is formulated for injection using appropriate vehicles such as saline, DMSO/PEG300/Tween-80 mixtures, or other biocompatible solvents. Endpoints include survival, inflammatory marker levels, histopathological scores, and quantitative lesion measurements.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of 5-Methoxy-DL-tryptophan are characteristic of a small molecule amino acid analog with moderate bioavailability. Following intraperitoneal administration, the compound is rapidly absorbed and distributed to tissues, with peak plasma concentrations typically achieved within 1-2 hours. The methoxy substitution at the 5-position of the indole ring confers metabolic stability, reducing the rapid degradation that compromises the reproducibility of assays using hydroxylated tryptophan analogs. The compound is primarily metabolized through hepatic pathways, with cytochrome P450 enzymes likely playing a role in its biotransformation. The elimination half-life in mice is estimated to be several hours, supporting twice-weekly dosing regimens in chronic studies. The compound's volume of distribution and protein binding characteristics have not been extensively characterized but are expected to be consistent with other amino acid derivatives. Renal clearance is likely the primary route of elimination for metabolites, as is typical for amino acid analogs. The pharmacokinetic profile of 5-MTP is formulation-dependent, with solubility in various vehicles affecting absorption rates and bioavailability. The compound is soluble in DMSO at 100 mg/mL and can be formulated for in vivo administration using combinations of DMSO, PEG300, Tween-80, and saline. Studies utilizing pegylated formulations of related compounds demonstrate that such modifications can extend half-life and decrease immunogenicity. The metabolic stability conferred by the methoxy group compared to hydroxylated analogs suggests that 5-MTP may have a more favorable pharmacokinetic profile for therapeutic development.
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| Toxicity/Toxicokinetics |
The toxicological profile of 5-Methoxy-DL-tryptophan has not been extensively characterized in formal toxicology studies, but available data suggest that the compound is well-tolerated at pharmacologically active doses. In mouse models of endotoxemia and atherosclerosis, repeated administration of 5-MTP at 23.4-23.5 mg/kg (i.p., twice weekly for up to 20 weeks) did not result in overt toxicity or adverse effects. The compound is an endogenous metabolite in mammals, suggesting that it is a naturally occurring substance with which the body is familiar. Unlike many synthetic anti-inflammatory agents, 5-MTP does not appear to cause gastrointestinal toxicity or immunosuppression at therapeutic doses. The methoxy substitution differentiates it from hydroxylated tryptophan analogs such as 5-HTP, which can have hormonal and sedative effects. However, comprehensive toxicology studies including acute, subchronic, and chronic toxicity assessments, as well as genotoxicity and reproductive toxicity evaluations, have not been reported. The compound is classified as a research chemical and is not approved for human use. Standard safety precautions should be observed when handling the compound, including the use of appropriate personal protective equipment. The absence of reported severe adverse effects in published studies suggests a favorable safety profile, but formal toxicological characterization would be required for clinical development. As with all research chemicals, the compound should be handled with care, and appropriate risk assessments should be conducted prior to in vivo use.
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| Additional Infomation |
5-Methoxy-DL-tryptophan is a type of tryptophan compound. Functionally, it is related to serotonin.
See also: 5-Methoxytryptophan (note moved to). 5-Methoxy-DL-tryptophan is a valuable research tool for studying inflammatory pathways, endothelial cell biology, and fibrotic disease mechanisms. It is particularly useful for investigating the distinct pharmacological profiles of tryptophan metabolites, as it offers a metabolically stable alternative to 5-hydroxytryptophan (5-HTP). The compound is compatible with in vitro macrophage assays, in vivo xenograft models, and a range of biochemical and cell-based studies. Its anti-inflammatory mechanism involving inhibition of COX-2 expression and p38 MAPK/NF-κB pathways distinguishes it from other tryptophan metabolites. The compound exhibits a more potent and specific anti-inflammatory profile compared to serotonin and melatonin, particularly in the context of COX-2 inhibition. 5-MTP has demonstrated efficacy in various models of tissue fibrosis, including in the lungs, kidneys, and liver, by inhibiting fibroblast-to-myofibroblast differentiation. It also exhibits protective effects on the vascular endothelium and can attenuate intimal hyperplasia following arterial injury. While the L-enantiomer is the primary biologically active form as the endogenous metabolite, the specific activity of the D-isomer remains an area for further investigation. The compound is not currently approved for any clinical indication and is strictly for research use only. Its unique mechanism of action, distinct from other tryptophan metabolites, positions it as a promising lead compound for further drug development in inflammatory, fibrotic, and cardiovascular diseases. |
| Molecular Formula |
C12H14N2O3
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|---|---|
| Molecular Weight |
234.25
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| Exact Mass |
234.1
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| CAS # |
28052-84-8
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| PubChem CID |
119802
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
478.3±45.0 °C at 760 mmHg
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| Melting Point |
258-261 °C (dec.)(lit.)
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| Flash Point |
243.1±28.7 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.663
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| LogP |
0.95
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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 |
4
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| Heavy Atom Count |
17
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| Complexity |
285
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC1=CC2=C(C=C1)NC=C2CC(C(=O)O)N
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| InChi Key |
KVNPSKDDJARYKK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C12H14N2O3/c1-17-8-2-3-11-9(5-8)7(6-14-11)4-10(13)12(15)16/h2-3,5-6,10,14H,4,13H2,1H3,(H,15,16)
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| Chemical Name |
2-amino-3-(5-methoxy-1H-indol-3-yl)propanoic 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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: 100 mg/mL (426.89 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 | 4.2689 mL | 21.3447 mL | 42.6894 mL | |
| 5 mM | 0.8538 mL | 4.2689 mL | 8.5379 mL | |
| 10 mM | 0.4269 mL | 2.1345 mL | 4.2689 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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