TriDAP dihydrochloride
Based on 1 publication(s) in Google Scholar
TriDAP dihydrochloride (L-Ala-γ-D-Glu-meso-diaminopimelic acid dihydrochloride) is a NOD1 agonist with a Kd value of 34.5 μM. TriDAP dihydrochloride enhances the binding of NOD1-RICK, promotes RICK phosphorylation, and activates the NF-κB, TAK1, MEK/ERK, p38 and interferon response pathways. TriDAP dihydrochloride downregulates Runx2 via increasing ubiquitination and reduces trabecular bone parameters. TriDAP dihydrochloride decreases IκBα levels and increases p65 levels. TriDAP dihydrochloride induces the secretion of proinflammatory mediators IL-8 and prostaglandins, triggers tissue inflammation and innate immune activation, and inhibits SARS-CoV-2 replication in lung epithelial cells. TriDAP dihydrochloride increases the RANKL/OPG ratio in mice, reduces bone mass and enhances osteoclast activity, and inhibits new bone formation by decreasing the mineralization deposition rate in mice. TriDAP dihydrochloride can be used in research related to pulpitis, chronic ulcerative colitis, Crohn's disease and SARS-CoV-2 infection.
Nos produits utilisent uniquement pour la recherche. Nous ne vendons pas aux patients.
- Pureté : 98.14%
- Formule: C15H28Cl2N4O8
- Masse moléculaire:463.31
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Stockage:
Sealed storage, away from moisture.
Powder -80°C, 2 years , -20°C, 1 year* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Activité biologique
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Chemical Information
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Solvant et solubilité
- Protocole
- Pureté et documentation
- Références
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Help & FAQs
Publications Citing Use of MedChemExpress (MCE) TriDAP dihydrochloride
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Activité biologique
Description
IC50 & Target
[4]|
hNOD1 34.5 μM (Kd) |
IL-8 |
In Vitro
TriDAP (0-10 μg/mL; 6 days) dihydrochloride dose-dependently inhibits osteogenic differentiation of primary mouse calvarial osteoblast precursors and enhances osteoclast differentiation in the co-culture system of mouse BMMs and primary calvarial osteoblast precursors[1].
TriDAP (0-10 μg/mL; 2 days) dihydrochloride downregulates the expression of osteogenic marker genes including Alp, Bsp and Runx2 in MC3T3-E1 mouse preosteoblasts in a dose-dependent manner[1].
TriDAP (0-10 μg/mL; 18 h) dihydrochloride dose-dependently inhibits the transcriptional activity of Runx2 in MC3T3-E1 mouse preosteoblasts[1].
TriDAP (1 μg/mL; 2-24 h) dihydrochloride reduces the stability of Runx2 protein in transiently transfected HEK293 cells. After 8 hours of co-incubation with Cycloheximide (HY-12320), the relative level of Runx2 decreases by approximately 50%, which enhances Smurf1-mediated Runx2 degradation in cells[1].
TriDAP (1-10 μg/mL; 24 h) dihydrochloride increases the ubiquitination level of Runx2 in transiently transfected HEK293 cells in a dose-dependent manner[1].
TriDAP (0.1-10 μg/mL; 2 days) dihydrochloride dose-dependently increases the RANKL/OPG ratio in primary mouse calvarial osteoprogenitor cells, through a mechanism that decreases OPG secretion and increases RANKL secretion[1].
TriDAP (1 μg/mL; 15-90 min) dihydrochloride activates the NF-κB pathway in MC3T3-E1 mouse osteoprogenitor cells by reducing IκBα levels and increasing p65 levels, and induces nuclear translocation of NF-κB p65[1].
TriDAP (0.1-10 μg/mL; 24 h) dihydrochloride dose-dependently enhances the transcriptional activity of NF-κB in MC3T3-E1 mouse preosteoblasts[1].
TriDAP (10 μg/mL; 10 days) dihydrochloride inhibits osteogenic mineralization of wild-type primary mouse calvarial preosteoblasts, but this effect is abolished in NOD1-deficient preosteoblasts, indicating that this action is dependent on NOD1[1].
TriDAP (10 μg/mL; 16 h) dihydrochloride preferentially activates the transcriptional activity of NF-κB in HEK293 cells transfected with NOD1, whereas the activity is extremely low in cells transfected with NOD2[1].
TriDAP (10-100 μM) dihydrochloride binds directly to purified full-length NOD1 protein, with a measured Kd value of 34.5 μM via SPR[4].
Binding of TriDAP (0.1 mM) dihydrochloride to purified full-length NOD1 protein increases the binding affinity between purified RICK protein and NOD1, with SPR assays showing that its Kd value decreases from 4.13 μM to 3.26 μM[4].
TriDAP (0-5 mM; 1-4 h) dihydrochloride induces the expression of NOD1 and secretion of IL-8 in Caco2-BBE intestinal epithelial cells, and this pro-inflammatory response depends on the expression of NOD1[4].
TriDAP dihydrochloride activates the NOD1 pathway in HEK-Blue hNOD1 cells, with an EC50 of 700 ng/mL[5].
TriDAP (0.4-50 µM; 6-24 h) dihydrochloride increases the proportion of IL-8+ human lung epithelial A549 cells by 3.29-fold, elevates the proportion of IL-8+ human lung epithelial A549-Dual cells, does not reduce cell viability, and activates the NF-κB and ISRE pathways in human lung epithelial A549-Dual cells[5].
TriDAP (50 µM; 8 h) dihydrochloride upregulates the mRNA expression levels of IL-8, CXCL10 and ISG15 in the human lung epithelial cell line A549-Dual[5].
TriDAP (2-50 µM; 3-hour pretreatment; 48 hours post-infection) dihydrochloride dose-dependently inhibits SARS-CoV-2 replication in human lung epithelial A549-Dual cells, with an EC50 of 7.75 µM, and reduces the number of infected cells by 49% at a concentration of 50 µM[5].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:primary murine calvarial osteoblast precursors
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Concentration:0.1, 1, 10 μg/mL
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Incubation Time:6 days
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Result:Inhibited osteoblast differentiation in a dose-dependent manner, as evidenced by reduced ALP staining intensity with increasing TriDAP concentrations.
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Cell Line:MC3T3-E1 murine osteoblast precursor cells
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Concentration:1, 10 μg/mL
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Incubation Time:2 days
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Result:Decreased the expression of osteogenic marker genes Alp, Bsp, and Runx2 in a dose-dependent manner compared to untreated cells.
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Cell Line:transiently transfected HEK293 cells
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Concentration:1 μg/mL
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Incubation Time:2, 4, 8 h
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Result:Decreased the half-life of Runx2 protein; reduced relative Runx2 levels significantly compared to untreated cells at 4 and 8 h.
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Cell Line:primary murine calvarial osteoblast precursors
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Concentration:0.1, 1, 10 μg/mL
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Incubation Time:2 days
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Result:Decreased OPG levels and increased RANKL levels in a dose-dependent manner, resulting in a dose-dependent increase in the RANKL/OPG ratio; caused statistically significant changes at all tested concentrations compared to untreated cells.
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Cell Line:MC3T3-E1 murine osteoblast precursor cells
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Concentration:1 μg/mL
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Incubation Time:15, 30, 60, 90 min
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Result:Decreased IκBα protein levels in a time-dependent manner and increased p65 protein levels over the same time course.
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Cell Line:MC3T3-E1 murine osteoblast precursor cells
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Concentration:1 μg/mL
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Incubation Time:15 min
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Result:Induced translocation of p65 from the cytoplasm to the nucleus within 15 minutes of treatment.
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Cell Line:human lung epithelial A549-Dual cells
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Concentration:50 µM
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Incubation Time:8 h
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Result:Induced a 1.68-fold increase in IL-8 mRNA compared to untreated control.
Significantly upregulated CXCL10 mRNA compared to untreated control.
Significantly upregulated ISG15 mRNA compared to untreated control.
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6 (6-week-old male)[1]
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Dosage:1.25 mg/kg
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Administration:i.p.; on days 0 and 4/ on days 1 and 5
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Result:Caused significant reductions in trabecular bone volume (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) compared to PBS controls.
Left trabecular separation (Tb.Sp) unchanged.
Confirmed decreased trabecular bone in distal femurs via H&E staining.
Increased TRAP-positive surface area on bone surfaces, indicating enhanced osteoclast activity.
Significantly increased the RANKL/OPG ratio in bone marrow extracellular fluids relative to PBS controls.
Resulted in a significant reduction in the calcein-labeled mineralized surface of distal femurs compared to PBS controls.
Caused a significant decrease in mineral apposition rate compared to PBS controls.
Chemical Information
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Appearance Solid
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Masse moléculaire 463.31
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Formule C15H28Cl2N4O8
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Color White to light yellow
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Synonyms
L-Ala-γ-D-Glu-meso-diaminopimelic acid dihydrochloride
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Sequence
Ala-{d-γGlu}-{meso-Diaminopimelic acid}
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Sequence Shortening
A-{d-γGlu}-{meso-Diaminopimelic acid}
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
Sealed storage, away from moisture
Powder -80°C 2 years -20°C 1 year * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Publications (1)
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Journal Impact Factor
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Most Recent
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Exploration
A Bioorthogonal and Programmable Bacterial Delivery System for Spatiotemporally Targeted Therapy of Solid Tumors. [Abstract]2025 Dec 18;5(6):20240396. PMID: 41476656
Solvant et solubilité
In Vitro:
DMSO : 100 mg/mL (215.84 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (5.40 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (5.40 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocole
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Kinase activity and phosphorylation assays
Kinase activity assays measure the ability of kinases to transfer phosphate groups from ATP to specific substrates, while phosphorylation assays detect the presence and levels of phosphorylated proteins. Common methods include radiolabeled ATP incorporation (e. g. ,), ADP release detection via bioluminescence (e. g. ,[3]), enzyme-linked immunosorbent assays (ELISA) for phospho-specific epitopes (e. g. ,[6]), and microtiter-based formats for high-throughput screening (e. g. ,[8]). The ADP-Glo assay quantifies kinase activity by measuring ADP produced during phosphorylation using a luciferase-based system. Radiometric assays involve autoradiography or scintillation counting after incorporation of 32P-labeled ATP into substrate proteins. ELISA-based approaches rely on phospho-specific antibodies to detect activated kinases in cell lysates or purified samples.
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Western Blot
Western blotting (WB) is a commonly used experimental method in molecular biology, biochemistry, and immunogenetics for identifying and quantifying target proteins. It combines gel electrophoresis with immunoassay, enabling researchers to analyze protein expression, post-translational modifications, and molecular weight.
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Research Protocol for Infectious Diseases
Infectious-disease experiments test how pathogens interact with host barriers, innate immune receptors, inflammatory signaling, pathogen replication, and tissue injury; pattern-recognition receptors such as TLRs, RIG-I-like receptors, NOD-like receptors, and inflammasomes detect microbial molecules and activate NF-κB, interferon, and cytokine responses. The central hypothesis is that infection severity reflects the balance between pathogen burden and host response: protective inflammation restricts pathogen growth, whereas excessive or mislocalized inflammation contributes to tissue damage and disease phenotype. Unresolved questions include which host pathways are protective versus pathogenic, why some infection models fail to translate to human disease, and which combined readouts best predict clinically relevant infection outcomes.
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Mesenchymal stromal/stem cell osteogenic differentiation
Mesenchymal stromal/stem cells can be induced toward an osteoblast-like lineage in vitro by culture in osteogenic medium containing dexamethasone, ascorbic acid or ascorbate-2-phosphate, and β-glycerophosphate; the differentiation process is commonly evaluated by alkaline phosphatase activity, osteogenic marker expression, collagenous matrix formation, and calcium-rich matrix mineralization. The main readouts are alkaline phosphatase activity as an early osteogenic marker and Alizarin Red S staining as a calcium-deposit readout for mineralized extracellular matrix; Alizarin Red S can be inspected microscopically or extracted and measured colorimetrically at 405 nm.
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DSS-Induced Colitis
Dextran sulfate sodium (DSS)-induced colitis is generated by administering DSS in mouse drinking water, producing epithelial injury, barrier disruption, weight loss, diarrhea, fecal blood, colon shortening, histologic mucosal damage, and inflammatory mediator changes; the model is mainly used to study acute or chronic intestinal inflammation resembling selected features of ulcerative colitis. DSS injury is interpreted through clinical and tissue readouts rather than a single molecular endpoint: daily body weight, stool consistency, and bleeding are combined into a disease activity index, while colon length, histology, cytokines, myeloperoxidase activity, intestinal permeability, and tight-junction markers provide complementary measures of inflammation and barrier damage.
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TNBS-Induced Colitis
TNBS-induced colitis is produced by intrarectal delivery of 2,4,6-trinitrobenzene sulfonic acid in ethanol, where ethanol disrupts the mucosal barrier and TNBS haptenates colonic proteins, generating immune-mediated colonic inflammation with weight loss, diarrhea, ulceration, transmural injury, inflammatory-cell infiltration, and cytokine responses. The model is used as an experimental intestinal inflammation model with Crohn’s disease–like features, especially when Th1-type responses, IL-12–dependent inflammation, chronic relapsing inflammation, or fibrosis-related endpoints are studied.
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Protocol for Kinase activity and phosphorylation assays
Kinase activity assays measure transfer of phosphate from ATP to a protein or peptide substrate, generating phosphorylated substrate, ADP, or incorporated radiolabeled phosphate as the readout; phosphorylation assays measure site-specific phosphorylation in cells or tissues as a proxy for kinase-pathway activation, inhibition, or substrate regulation. Phosphorylation can be detected by phospho-specific Western blot, immunoprecipitation kinase assay, phospho-immunofluorescence, phospho-flow cytometry, luminescent ADP detection, radiolabeled ATP incorporation, or reporter-based pathway assays, and these readouts can be applied to cancer cells, primary neurons, mouse tumors, organoids, inflammatory macrophages, ferroptosis studies, and mitophagy studies when the kinase target is biologically relevant.
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Research Protocol for Inflammation-related Diseases
The NLRP3 inflammasome is a cytosolic innate immune signaling platform that integrates priming signals and danger-signal activation to promote caspase-1 activation, maturation of IL-1β and IL-18, and gasdermin D-mediated pyroptotic cell death. The core experimental logic is to determine whether inflammatory disease phenotypes are driven by increased NLRP3 expression, ASC-containing inflammasome assembly, caspase-1 cleavage, GSDMD cleavage, and extracellular release of IL-1β/IL-18 rather than by nonspecific cell injury alone. The pathway is strongly linked to inflammation-related disease phenotypes because monosodium urate crystals activate NALP3/NLRP3 inflammasome signaling in gout-like crystal inflammation, cholesterol crystals activate NLRP3 inflammasomes in atherogenesis models, and DSS-induced intestinal inflammation has been reported to involve NLRP3 inflammasome activity. However, experimental colitis studies also show context-dependent protective effects of NLRP3 inflammasome co
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Osteoclast differentiation from monocyte/macrophage precursors
Osteoclast differentiation is an in vitro induction assay in which monocyte/macrophage-lineage precursors are exposed to macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL), generating multinucleated osteoclasts that are commonly identified by tartrate-resistant acid phosphatase (TRAP) staining and functionally confirmed by resorption pits on dentin, bone, or mineralized substrates. M-CSF supports survival and expansion of osteoclast precursors, while RANKL binding to RANK drives osteoclast commitment, fusion, maturation, and resorptive function; osteoprotegerin inhibits this pathway by binding RANKL and preventing RANK activation. The main readouts are the number of TRAP-positive multinucleated cells, formation of F-actin rings, and resorbed surface area; TRAP-positive multinucleated cells indicate osteoclast differentiation, whereas pit formation on dentin, bone, or mineralized coating indicates functional bone-resorbing activity.
Pureté et documentation
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Fiche technique (290 KB)
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SDS (254 KB)
- English - EN (254 KB)
- Français - FR (254 KB)
- Deutsch - DE (254 KB)
- Norwegian - NO (254 KB)
- Español - ES (254 KB)
- Swedish - SV (254 KB)
- Italian - IT (254 KB)
- Korean - KR (254 KB)
- Portuguese - PT (254 KB)
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Instruction de manipulation (2659 KB)
Références
[1]. Park OJ, et al. The dual roles of peptidoglycans: NOD1 and NOD2 inversely regulate bone metabolism. Exp Mol Med. 2025;57(8):1837-1846. [Content Brief]
[2]. Enoksson M, et al. Human cord blood-derived mast cells are activated by the Nod1 agonist M-TriDAP to release pro-inflammatory cytokines and chemokines. J Innate Immun. 2011;3(2):142-149. [Content Brief]
[3]. Chang MC, et al. Inducing phospholipase A2 and cyclooxygenase-2 expression and prostaglandins' production of human dental pulp cells by activation of NOD receptor and its downstream signaling. Int J Biol Macromol. 2025;292:139193. [Content Brief]
[4]. Laroui H, et al. L-Ala-γ-D-Glu-meso-diaminopimelic acid (DAP) interacts directly with leucine-rich region domain of nucleotide-binding oligomerization domain 1, increasing phosphorylation activity of receptor-interacting serine/threonine-protein kinase 2 and its interaction with nucleotide-binding oligomerization domain 1. J Biol Chem. 2011;286(35):31003-31013. [Content Brief]
[5]. Garcia-Vidal E, et al. Nucleotide-Binding Oligomerization Domain 1 (NOD1) Agonists Prevent SARS-CoV-2 Infection in Human Lung Epithelial Cells through Harnessing the Innate Immune Response. Int J Mol Sci. 2024;25(10):5318. Published 2024 May 13. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.1584 mL | 10.7919 mL | 21.5838 mL | 53.9596 mL |
| 5 mM | 0.4317 mL | 2.1584 mL | 4.3168 mL | 10.7919 mL | |
| 10 mM | 0.2158 mL | 1.0792 mL | 2.1584 mL | 5.3960 mL | |
| 15 mM | 0.1439 mL | 0.7195 mL | 1.4389 mL | 3.5973 mL | |
| 20 mM | 0.1079 mL | 0.5396 mL | 1.0792 mL | 2.6980 mL | |
| 25 mM | 0.0863 mL | 0.4317 mL | 0.8634 mL | 2.1584 mL | |
| 30 mM | 0.0719 mL | 0.3597 mL | 0.7195 mL | 1.7987 mL | |
| 40 mM | 0.0540 mL | 0.2698 mL | 0.5396 mL | 1.3490 mL | |
| 50 mM | 0.0432 mL | 0.2158 mL | 0.4317 mL | 1.0792 mL | |
| 60 mM | 0.0360 mL | 0.1799 mL | 0.3597 mL | 0.8993 mL | |
| 80 mM | 0.0270 mL | 0.1349 mL | 0.2698 mL | 0.6745 mL | |
| 100 mM | 0.0216 mL | 0.1079 mL | 0.2158 mL | 0.5396 mL |
TriDAP dihydrochloride Related Classifications
HY-P5522A Related Classifications
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