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]

LDN193189 (GMP)（250 nM；0-7 天）刺激人多能干细胞 (hPSC) 直接发育为中脑多巴胺神经元 (mDA)[1]。在体外，浓度为 200 nM 的 LDN193189 (GMP) 可诱导 hPSC 产生葡萄糖反应性 β 细胞 [2]。

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在BMP4过表达的人肺腺癌细胞（A549）中，LDN-193189 (DM-3189)（1 μM）处理24小时后，抑制BMP介导的Smad1/5/8磷酸化88%。它在mRNA水平下调BMP靶基因（ID1降低75%；ID2降低70%；ID3降低68%），72小时后MTT法检测显示抑制细胞增殖62% [3]
- 在人肝癌细胞（HepG2）中，LDN-193189 (DM-3189)（2 μM）处理48小时后诱导G2/M期细胞周期阻滞（G2/M期细胞比例从22%升至45%），72小时后诱导凋亡，膜联蛋白V阳性细胞比例从6%升至38%。它上调裂解型胱天蛋白酶-3（3.5倍），下调周期蛋白B1（降低65%）[4]
- 在经BMP2处理的小鼠胚胎成纤维细胞（MEFs）中，LDN-193189 (DM-3189)（0.5 μM）阻断BMP诱导的成骨分化，7天后碱性磷酸酶（ALP）活性降低70%，14天后矿化结节形成减少65% [1]
- 在正常人支气管上皮细胞（HBECs）中，LDN-193189 (DM-3189) 在浓度高达20 μM时毒性较低（细胞活力较对照组>85%）[3]

LDN-193189（腹腔注射；3 mg/kg；每天；35 天）可能会影响乳腺癌细胞与其周围骨骼的相互作用[3]。 LDN-193189（腹腔注射；3 mg/kg；单剂量）可减少功能损伤和异位骨化 [1]。

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在荷皮下A549肺癌异种移植瘤的裸鼠中，口服 LDN-193189 (DM-3189)（25 mg/kg/天，持续28天）显著抑制肿瘤生长。与溶媒处理组相比，肿瘤体积减少63%，肿瘤重量降低58%。肿瘤组织中p-Smad1/5/8（降低72%）和Ki-67（降低55%）表达下调 [3]
- 在BMP2植入诱导的小鼠异位骨化模型中，腹腔注射 LDN-193189 (DM-3189)（10 mg/kg/天，持续14天）减少异位骨形成70%（微计算机断层扫描分析）。它抑制骨化组织中Smad1/5/8磷酸化，成骨标志物（Runx2降低65%；OCN降低60%）表达下调 [1]
- 在四氯化碳（CCl₄）诱导的大鼠肝纤维化模型中，口服 LDN-193189 (DM-3189)（15 mg/kg/天，持续8周）减少肝脏胶原蛋白含量55%，α-SMA阳性肌成纤维细胞数量减少62%。它抑制肝脏组织中的BMP/Smad信号，p-Smad1/5/8水平降低68% [4]

碱性磷酸酶活性[1] 我们将C2C12细胞以每孔2000个细胞的速度接种到96孔板中，在补充有2%FBS的DMEM中。我们用BMP配体和LDN-193189或载体以四硅酸盐的形式处理孔。我们在50μl Tris缓冲盐水和1%Triton X-100中培养6天后收集细胞。我们将裂解物加入96孔板中的对硝基-苯基磷酸盐试剂中1小时，然后评估碱性磷酸酶活性（405nm处的吸光度）。我们使用与用于碱性磷酸酶测量的重复孔相同处理的重复孔，通过cell Titer Water One（在490nm处的吸光度）测量细胞活力和数量。

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ALK2/ALK3激酶活性实验：将纯化的重组人ALK2或ALK3与Smad1衍生底物肽和 LDN-193189 (DM-3189)（0.1 nM-100 nM）在实验缓冲液（50 mM Tris-HCl，pH 7.5，10 mM MgCl₂，1 mM DTT，0.1 mM ATP）中于30°C孵育60分钟。通过放射性标记ATP计数检测磷酸化底物，从剂量-效应曲线计算IC50值 [1][2]
- ATP竞争性结合实验：将ALK2与递增浓度的ATP（0.05-1 mM）和固定浓度的 LDN-193189 (DM-3189)（5 nM）孵育。检测激酶活性以证实其与ALK2的ATP结合口袋竞争性结合 [2]
- 激酶选择性实验：采用各自的底物肽和实验缓冲液，将 LDN-193189 (DM-3189)（10 μM）对40+种激酶（包括ALK1、ALK4-6、PKA、PKC、ERK1/2）进行筛选。比色法定量激酶活性，未观察到对脱靶激酶的显著抑制（活性降低>50%）[2][3]

细胞培养[1] 如前所述，我们从野生型和CAG-Z-EGFP-caALK2转基因小鼠中分离出PASMC，并将其在补充有10%FBS的RPMI培养基中培养。我们通过用Ad.Cre（50的多重感染）或Ad.GFP作为对照感染，然后培养3天并传代，在体外诱导表达条件caALK2的PASMC的重组。我们在补充谷氨酰胺和10%FBS的DMEM中培养C2C12肌成纤维细胞（美国典型培养物保藏中心）。我们用药物抑制剂预培养细胞10分钟，然后在37°C下将其暴露于BMP4、TGF-β或血小板衍生生长因子BB配体30分钟。[1]

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肺癌细胞增殖与BMP信号实验：A549细胞以2×10⁵个/孔接种到6孔板中，用 LDN-193189 (DM-3189)（0.1-5 μM）预处理1小时，再用BMP4（10 ng/mL）刺激24-72小时。Western blot检测p-Smad1/5/8和总Smad1；qPCR分析ID1/ID2/ID3 mRNA水平；MTT法检测细胞活力 [3]
- 肝癌细胞周期与凋亡实验：HepG2细胞分别以3×10³个/孔（增殖实验）或2×10⁵个/孔（周期/凋亡实验）接种到96孔板或6孔板中。用 LDN-193189 (DM-3189)（0.5-5 μM）处理48-72小时。碘化丙啶染色流式细胞术分析细胞周期；膜联蛋白V-FITC/PI染色定量凋亡；Western blot检测裂解型胱天蛋白酶-3和周期蛋白B1 [4]
- MEF成骨分化实验：小鼠胚胎成纤维细胞以1×10⁵个/孔接种到6孔板中，在含BMP2（50 ng/mL）和 LDN-193189 (DM-3189)（0.1-2 μM）的成骨培养基中培养。7天比色法检测ALP活性；14天茜素红S染色定量矿化结节 [1]

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.

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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.

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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]

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]

[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.

4-[6-[4-(1-piperazinyl)phenyl]-3-pyrazolo[1,5-a]pyrimidinyl]quinoline is a member of pyrimidines.

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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]

C25H22N6

406.48

406.19

C, 73.87; H, 5.46; N, 20.67

1062368-24-4

LDN193189;1062368-24-4

25195294

Typically exists as light yellow to yellow solids at room temperature

1.3±0.1 g/cm3

1.740

1.81

58.35

1

5

3

31

587

0

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]

CDOVNWNANFFLFJ-UHFFFAOYSA-N

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

4-[6-(4-Piperazin-1-yl-phenyl)-pyrazolo[1,5-a]pyrimidin-3-yl]-quinoline

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

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 1.2 mg/mL (2.95 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 12.0 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 1.2 mg/mL (2.95 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 12.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: Saline (suspension): 30 mg/mL

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。