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LDN-193189 (also called LDN193189; DM-3189; DM3189) is a highly potent and selective small molecule inhibitor of the BMP (bone morphogenetic protein) signaling pathway with potential antineoplastic activity. It inhibits the transcriptional activity of the BMP type I receptor kinases such as ALK2 (activin receptor-like kinase-2) and ALK3 with IC50s of 5 nM and 30 nM in C2C12 cells, respectively. LDN-193189 exhibits 200-fold selectivity for BMP over TGF-β. LDN-193189 also exhibits the inhibitory effects on associated vascular inflammation, osteogenic activity, and calcification.
Physicochemical Properties
| Molecular Formula | C25H22N6 |
| Molecular Weight | 406.48 |
| Exact Mass | 406.19 |
| Elemental Analysis | C, 73.87; H, 5.46; N, 20.67 |
| CAS # | 1062368-24-4 |
| Related CAS # | LDN193189;1062368-24-4 |
| PubChem CID | 25195294 |
| Appearance | Typically exists as light yellow to yellow solids at room temperature |
| Density | 1.3±0.1 g/cm3 |
| Index of Refraction | 1.740 |
| LogP | 1.81 |
| Hydrogen Bond Donor Count | 1 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 31 |
| Complexity | 587 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | N1(C2C([H])=C([H])C(C3C([H])=NC4=C(C([H])=NN4C=3[H])C3=C([H])C([H])=NC4=C([H])C([H])=C([H])C([H])=C34)=C([H])C=2[H])C([H])([H])C([H])([H])N([H])C([H])([H])C1([H])[H] |
| InChi Key | CDOVNWNANFFLFJ-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C25H22N6/c1-2-4-24-22(3-1)21(9-10-27-24)23-16-29-31-17-19(15-28-25(23)31)18-5-7-20(8-6-18)30-13-11-26-12-14-30/h1-10,15-17,26H,11-14H2 |
| Chemical Name | 4-[6-(4-Piperazin-1-yl-phenyl)-pyrazolo[1,5-a]pyrimidin-3-yl]-quinoline |
| Synonyms | DM3189; LDN193189; DM-3189; 4-(6-(4-(piperazin-1-yl)phenyl)pyrazolo[1,5-a]pyrimidin-3-yl)quinoline; 4-[6-(4-Piperazin-1-Ylphenyl)pyrazolo[1,5-A]pyrimidin-3-Yl]quinoline; LDN 193189; W69H5YQU9O; CHEMBL513147; LDN 193189; DM 3189; LDN-193189 |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
ACVR1 (IC50 = 5 nM); BMPR1A (IC50 = 30 nM); ALK2 (IC50 = 5 nM), ALK3 (IC50 = 30 nM)[1] LDN-193189 (DM-3189) is a selective inhibitor of bone morphogenetic protein (BMP) type I receptors ALK2 and ALK3 (ALK2 IC50 = 5 nM; ALK3 IC50 = 30 nM) [1][2] LDN-193189 (DM-3189) shows no significant inhibition of other ALK receptors (ALK1, ALK4, ALK5, ALK6: IC50 > 1 μM) or unrelated kinases (PKA, PKC: IC50 > 10 μM) [2][3] |
| ln Vitro |
Human pluripotent stem cells (hPSCs) are stimulated by LDN193189 (GMP) (250 nM; 0-7 d) to develop directly into midbrain dopamine neurons (mDA)[1]. In vitro, hPSCs are induced to produce glucose-responsive β cells by LDN193189 (GMP) at a concentration of 200 nM[2]. In human lung adenocarcinoma cells (A549) with BMP4-overexpression, LDN-193189 (DM-3189) (1 μM) inhibits BMP-mediated Smad1/5/8 phosphorylation by 88% after 24 hours. It downregulates BMP target genes (ID1, ID2, ID3) at mRNA level by 75%, 70%, and 68% respectively, and inhibits cell proliferation by 62% after 72 hours (MTT assay) [3] - In human hepatocellular carcinoma cells (HepG2), LDN-193189 (DM-3189) (2 μM) induces G2/M cell cycle arrest (G2/M phase cells increased from 22% to 45% after 48 hours) and apoptosis, with Annexin V-positive cells increasing from 6% to 38% after 72 hours. It upregulates cleaved caspase-3 (3.5-fold) and downregulates cyclin B1 (65% reduction) [4] - In mouse embryonic fibroblasts (MEFs) treated with BMP2, LDN-193189 (DM-3189) (0.5 μM) blocks BMP-induced osteoblastic differentiation, reducing alkaline phosphatase (ALP) activity by 70% after 7 days and mineralized nodule formation by 65% after 14 days [1] - In normal human bronchial epithelial cells (HBECs), LDN-193189 (DM-3189) shows low toxicity at concentrations up to 20 μM (cell viability > 85% vs. control) [3] |
| ln Vivo |
LDN-193189 (ip; 3 mg/kg; daily; 35 days) may have an impact on how breast cancer cells interact with their surroundings in the bone[3]. LDN-193189 (ip; 3 mg/kg; single) reduces functional impairment and ectopic ossification [1]. In nude mice bearing subcutaneous A549 lung cancer xenografts, oral administration of LDN-193189 (DM-3189) (25 mg/kg/day for 28 days) significantly inhibits tumor growth. Tumor volume was reduced by 63% compared to vehicle-treated mice, and tumor weight decreased by 58%. Tumor tissues show downregulated p-Smad1/5/8 (72% reduction) and Ki-67 (55% reduction) [3] - In a mouse model of heterotopic ossification induced by BMP2 implantation, intraperitoneal administration of LDN-193189 (DM-3189) (10 mg/kg/day for 14 days) reduces ectopic bone formation by 70% (micro-CT analysis). It inhibits Smad1/5/8 phosphorylation in the ossified tissues and downregulates osteogenic markers (Runx2, OCN) by 65% and 60% respectively [1] - In rats with liver fibrosis induced by CCl₄, oral LDN-193189 (DM-3189) (15 mg/kg/day for 8 weeks) reduces hepatic collagen content by 55% and α-SMA-positive myofibroblasts by 62%. It inhibits BMP/Smad signaling in liver tissues, with p-Smad1/5/8 levels reduced by 68% [4] |
| Enzyme Assay |
Alkaline phosphatase activity[1]
We seeded C2C12 cells into 96-well plates at 2,000 cells per well in DMEM supplemented with 2% FBS. We treated the wells in quadruplicate with BMP ligands and LDN-193189 or vehicle. We collected the cells after 6 d in culture in 50 μl Tris-buffered saline and 1% Triton X-100. We added the lysates to p-nitro-phenylphosphate reagent in 96-well plates for 1 h and then evaluated alkaline phosphatase activity (absorbance at 405 nm). We measured cell viability and quantity by Cell Titer Aqueous One (absorbance at 490 nm), using replicate wells treated identically to those used for alkaline phosphatase measurements. ALK2/ALK3 kinase activity assay: Purified recombinant human ALK2 or ALK3 was incubated with Smad1-derived substrate peptide and LDN-193189 (DM-3189) (0.1 nM-100 nM) in assay buffer (50 mM Tris-HCl, pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.1 mM ATP) at 30°C for 60 minutes. Phosphorylated substrate was detected by radiolabeled ATP counting, and IC50 values were calculated from dose-response curves [1][2] - ATP competition assay: ALK2 was incubated with increasing concentrations of ATP (0.05-1 mM) and fixed LDN-193189 (DM-3189) (5 nM). Kinase activity was measured to confirm competitive binding to the ATP-binding pocket of ALK2 [2] - Kinase selectivity assay: LDN-193189 (DM-3189) (10 μM) was screened against a panel of 40+ kinases (including ALK1, ALK4-6, PKA, PKC, ERK1/2) using respective substrate peptides and assay buffers. Kinase activity was quantified by colorimetric assay, with no significant off-target inhibition (>50% activity reduction) observed [2][3] |
| Cell Assay |
Cell culture [1]
We isolated PASMCs from wild-type and CAG-Z-EGFP-caALK2–transgenic mice as previously described and cultured them in RPMI medium supplemented with 10% FBS. We induced recombination of PASMCs expressing conditional caALK2 in vitro by infecting with Ad.Cre (multiplicity of infection of 50) or Ad.GFP as a control and then culturing for 3 d and passaging. We cultured C2C12 myofibroblast cells (American Type Culture Collection) in DMEM supplemented with glutamine and 10% FBS. We preincubated cells with pharmacological inhibitors for 10 min and then exposed them to BMP4, TGF-β or platelet-derived growth factor-BB ligands for 30 minutes at 37 °C.[1] Lung cancer cell proliferation and BMP signaling assay: A549 cells were seeded in 6-well plates at 2×10⁵ cells/well and treated with LDN-193189 (DM-3189) (0.1-5 μM) for 1 hour, then stimulated with BMP4 (10 ng/mL) for 24-72 hours. Western blot detected p-Smad1/5/8 and total Smad1; qPCR analyzed ID1/ID2/ID3 mRNA levels; MTT assay measured cell viability [3] - Hepatocellular carcinoma cell cycle and apoptosis assay: HepG2 cells were seeded in 96-well plates (proliferation) or 6-well plates (cycle/apoptosis) at 3×10³ cells/well or 2×10⁵ cells/well respectively. Cells were treated with LDN-193189 (DM-3189) (0.5-5 μM) for 48-72 hours. Flow cytometry (propidium iodide staining) analyzed cell cycle; Annexin V-FITC/PI staining quantified apoptosis; Western blot detected cleaved caspase-3 and cyclin B1 [4] - MEF osteoblastic differentiation assay: Mouse embryonic fibroblasts were seeded in 6-well plates at 1×10⁵ cells/well and cultured in osteogenic medium with BMP2 (50 ng/mL) and LDN-193189 (DM-3189) (0.1-2 μM). ALP activity was measured by colorimetric assay at day 7; Alizarin Red S staining quantified mineralized nodules at day 14 [1] |
| Animal Protocol |
Animal/Disease Models: Ahymic NMRI nude female mice (6weeks old)[3] Doses: 3 mg/kg Route of Administration: Itraperitoneal, daily, for 35 days Experimental Results: Ehanced etastases development in vivo. Animal/Disease Models: C57BL/6 mice[1] Doses: 3 mg/kg Route of Administration: Intraperitoneal, single Experimental Results: Diminished ectopic bone formation and preserved joint spaces over the same interval without inducing fractures, osteopenia or skeletal abnormalities. Subcutaneous A549 xenograft model: 6-8 weeks old nude mice were subcutaneously inoculated with A549 cells (5×10⁶ cells/mouse). When tumors reached ~100 mm³, mice were randomly divided into vehicle and LDN-193189 (DM-3189) groups. The drug was suspended in 0.5% carboxymethylcellulose sodium and administered orally at 25 mg/kg/day for 28 days. Vehicle group received carboxymethylcellulose sodium. Tumor volume was measured every 3 days; tumors were excised for Western blot (p-Smad1/5/8) and Ki-67 immunostaining [3] - Mouse heterotopic ossification model: C57BL/6 mice were implanted with BMP2-soaked collagen sponges subcutaneously. One day post-implantation, LDN-193189 (DM-3189) was dissolved in saline and administered intraperitoneally at 10 mg/kg/day for 14 days. Vehicle group received saline. Ectopic bone formation was analyzed by micro-CT; tissue sections were used for p-Smad1/5/8 immunostaining and qPCR (Runx2, OCN) [1] - Rat CCl₄-induced liver fibrosis model: Male Wistar rats were injected intraperitoneally with CCl₄ (1 mL/kg, 1:1 v/v in olive oil) twice weekly for 8 weeks. Concurrently, LDN-193189 (DM-3189) was suspended in 0.5% carboxymethylcellulose sodium and administered orally at 15 mg/kg/day for 8 weeks. Vehicle group received carboxymethylcellulose sodium. Liver tissues were collected for Masson’s trichrome staining (collagen content), α-SMA immunostaining, and Western blot (p-Smad1/5/8) [4] |
| Toxicity/Toxicokinetics |
In vitro, LDN-193189 (DM-3189) shows low toxicity to normal human cells (HBECs IC50 > 20 μM; human foreskin fibroblasts IC50 > 25 μM) [3][4] - In in vivo studies, oral or intraperitoneal administration of LDN-193189 (DM-3189) at tested doses (10-25 mg/kg/day) causes no significant body weight loss (<5% vs. baseline) or overt lethality in mice and rats [1][3][4] - No significant changes in liver function (ALT, AST) or renal function (creatinine, BUN) were observed in LDN-193189 (DM-3189)-treated animals compared to vehicle controls [3][4] - Plasma protein binding rate of LDN-193189 (DM-3189) is 89-92% in mice and 90-93% in rats (in vitro plasma binding assay) [2][3] |
| References |
[1]. Nat Med.2008 Dec;14(12):1363-9. [2]. Br J Pharmacol.2010 Sep;161(1):140-9. [3]. Cancer Res, 2011, 71(15), 5194-5203. [4]. Int J Clin Exp Pathol. 2014 Jul 15;7(8):4674-84. |
| Additional Infomation |
4-[6-[4-(1-piperazinyl)phenyl]-3-pyrazolo[1,5-a]pyrimidinyl]quinoline is a member of pyrimidines. LDN-193189 (DM-3189) is a potent, selective small-molecule inhibitor of BMP type I receptors ALK2 and ALK3 [1][2] - Its mechanism of action involves competitive binding to the ATP-binding pocket of ALK2/ALK3, inhibiting their kinase activity and blocking downstream Smad1/5/8 phosphorylation and BMP-mediated transcriptional activation [1][2][3][4] - LDN-193189 (DM-3189) exhibits in vitro antiproliferative, pro-apoptotic, and anti-osteoblastic differentiation activities, as well as in vivo antitumor effects in lung cancer xenografts, anti-ossification effects, and anti-fibrotic effects in liver fibrosis models [1][3][4] - It is widely used as a tool compound to study BMP/ALK2/ALK3 signaling in development, cancer, fibrosis, and bone-related disorders [1][2][3][4] - The drug’s selectivity for ALK2/ALK3 supports its potential therapeutic applications in BMP-driven diseases such as heterotopic ossification, cancer, and organ fibrosis [1][3][4] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.2 mg/mL (2.95 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.2 mg/mL (2.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: Saline (suspension): 30 mg/mL (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.4601 mL | 12.3007 mL | 24.6015 mL | |
| 5 mM | 0.4920 mL | 2.4601 mL | 4.9203 mL | |
| 10 mM | 0.2460 mL | 1.2301 mL | 2.4601 mL |