TriDAP dihydrochloride  (Synonyms: L-Ala-γ-D-Glu-meso-diaminopimelic acid dihydrochloride)

TriDAP dihydrochloride (L-Ala-γ-D-Glu-meso-diaminopimelic acid dihydrochloride) is a NOD1 agonist with a K_d 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^{[1][2][3][4][5]}.

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 K_d 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 K_d 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.

TriDAP (1.25 mg/kg; i.p.; administered on Day 0 and Day 4 / Day 1 and Day 5) dihydrochloride reduces bone mass and enhances osteoclast activity by increasing the RANKL/OPG ratio in male C57BL/6 mice, and inhibits new bone formation by decreasing the mineralization apposition rate in C57BL/6 mice^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

463.31

C₁₅H₂₈Cl₂N₄O₈

Solid

White to light yellow

Ala-{d-γGlu}-{meso-Diaminopimelic acid}

A-{d-γGlu}-{meso-Diaminopimelic acid}

Room temperature in continental US; may vary elsewhere.

Sealed storage, away from moisture

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

* 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.

• [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.

  [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.

  [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.