VEGF/VEGFR2
Tanshinone IIA (Dan Shen ketone) targets vascular endothelial growth factor receptor 2 (VEGFR2) (IC50 = 0.25 μM) [2]
Tanshinone IIA (Dan Shen ketone) targets signal transducer and activator of transcription 3 (STAT3) [1]
Tanshinone IIA (Dan Shen ketone) targets nuclear factor-κB (NF-κB) signaling pathway [1,4]
Tanshinone IIA (Dan Shen ketone) targets mitogen-activated protein kinase (MAPK) pathway [3]

Tanshinone IIA (Tanshinone B) is the most prevalent diterpene quinone in Danshen, where Salviae miltiorrhizae Radix is a commonly prescribed traditional herbal remedy used to treat inflammatory and cardiovascular conditions. Heart myocytes are shielded from oxidative stress-induced apoptosis by tanshinone IIA. The prevention of lipid peroxidation, enhanced scavenging of oxygen free radicals, and upregulation of the Bcl-2/Bax ratio all contribute to the in vivo protection.[1] It has recently been shown that tanshinone IIA can cause apoptosis and cell death in a range of tumor types.[2]

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In human hepatocellular carcinoma HepG2 cells, Tanshinone IIA (Dan Shen ketone) (2.5–20 μM) dose-dependently inhibited cell proliferation, with an IC50 of ~8 μM after 72 hours. It induced apoptosis: Annexin V-FITC/PI staining showed apoptotic rate increased to ~55% at 15 μM, accompanied by upregulated cleaved caspase-3 and Bax, downregulated Bcl-2 [1]
- In human lung cancer A549 cells, Tanshinone IIA (Dan Shen ketone) (1–10 μM) suppressed VEGFR2 phosphorylation and downstream signaling: at 5 μM, p-VEGFR2, p-AKT, and p-ERK1/2 levels decreased by ~60%, ~55%, and ~52%, respectively. It inhibited cell migration (Transwell assay) by ~65% and tube formation by ~70% at 5 μM [2]
- In lipopolysaccharide (LPS)-stimulated BV2 microglia, Tanshinone IIA (Dan Shen ketone) (1–10 μM) dose-dependently reduced inflammatory cytokine production: at 8 μM, TNF-α, IL-6, and iNOS mRNA levels decreased by ~68%, ~62%, and ~70%, respectively. It inhibited NF-κB p65 nuclear translocation by ~65% at 8 μM [4]
- In oxygen-glucose deprivation/reoxygenation (OGD/R)-induced PC12 cells, Tanshinone IIA (Dan Shen ketone) (0.5–4 μM) protected against cell injury: at 2 μM, cell viability increased from ~45% to ~78%, intracellular reactive oxygen species (ROS) production reduced by ~60%, and p-ERK1/2, p-JNK levels regulated by ~50–55% [3]
- In human colon cancer HT29 cells, Tanshinone IIA (Dan Shen ketone) (5–20 μM) arrested cells at G0/G1 phase: G0/G1 phase proportion increased from ~50% to ~72% at 15 μM, with downregulated cyclin D1 and CDK4 protein levels [1]

LD50: Rats 25g/kg (i.g.) [3]

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In BALB/c nude mouse xenograft model of human lung cancer (A549 cells), intraperitoneal administration of Tanshinone IIA (Dan Shen ketone) (10, 20 mg/kg/3 days for 4 weeks) significantly inhibited tumor growth. Tumor volume was reduced by ~48% (10 mg/kg) and ~68% (20 mg/kg), tumor weight decreased by ~45% (10 mg/kg) and ~65% (20 mg/kg) compared to control. Tumor tissues showed reduced microvessel density (by ~60% at 20 mg/kg) and p-VEGFR2 expression [2]
- In LPS-induced neuroinflammation mice, intraperitoneal injection of Tanshinone IIA (Dan Shen ketone) (5, 10 mg/kg/day for 7 days) reduced brain inflammatory responses: 10 mg/kg group showed TNF-α and IL-6 levels in cerebral cortex decreased by ~62% and ~58%, respectively. Microglia activation was inhibited by ~65% (10 mg/kg) [4]
- In middle cerebral artery occlusion (MCAO)-induced cerebral ischemia rats, intravenous administration of Tanshinone IIA (Dan Shen ketone) (5 mg/kg) 2 hours after ischemia reduced infarct volume by ~55% and improved neurological deficit scores (from ~3.5 to ~1.8). It enhanced antioxidant capacity in brain tissues: SOD activity increased by ~2.1-fold, MDA content reduced by ~60% [3]

VEGFR2 kinase activity assay: Recombinant VEGFR2 kinase domain was incubated with Tanshinone IIA (Dan Shen ketone) (0.05–5 μM) in reaction buffer containing ATP and a specific peptide substrate. The mixture was incubated at 30°C for 40 minutes, and phosphorylated substrate was detected by ELISA. The inhibition rate of VEGFR2 kinase activity was calculated, and IC50 value was determined from dose-response curves [2]
- NF-κB DNA-binding activity assay: LPS-stimulated BV2 cell nuclear extracts were prepared and incubated with Tanshinone IIA (Dan Shen ketone) (1–10 μM) for 30 minutes. A biotin-labeled NF-κB consensus oligonucleotide was added, and electrophoretic mobility shift assay (EMSA) was performed. The binding intensity of NF-κB to DNA was quantified by densitometry [4]
- MAPK kinase activity assay: OGD/R-induced PC12 cell lysates were immunoprecipitated with ERK1/2 or JNK antibodies. The immunocomplexes were incubated with Tanshinone IIA (Dan Shen ketone) (0.5–4 μM), ATP, and recombinant substrate at 30°C for 30 minutes. Phosphorylated substrate was detected by Western blot, and kinase activity was quantified [3]

Hepatocellular carcinoma cell proliferation and apoptosis assay: HepG2 cells were seeded in 96-well plates (5×103 cells/well) and treated with Tanshinone IIA (Dan Shen ketone) (2.5–20 μM) for 24–72 hours. MTT assay measured cell viability to calculate IC50. Apoptosis was detected by Annexin V-FITC/PI staining and flow cytometry. Western blot analyzed cleaved caspase-3, Bax, Bcl-2, cyclin D1, and CDK4 [1]
- Lung cancer cell and angiogenesis-related assay: A549 cells were treated with Tanshinone IIA (Dan Shen ketone) (1–10 μM) for 24–48 hours. Western blot detected p-VEGFR2, p-AKT, p-ERK1/2. Transwell assay evaluated cell migration (cells seeded in upper chambers with drug, stained and counted after 24 hours). Tube formation assay: HUVECs were seeded on Matrigel with drug, tube structures analyzed after 6 hours [2]
- Microglia inflammatory response assay: BV2 cells were stimulated with LPS (1 μg/mL) and Tanshinone IIA (Dan Shen ketone) (1–10 μM) for 24 hours. RT-PCR quantified TNF-α, IL-6, iNOS mRNA. Nuclear extracts were prepared for NF-κB EMSA [4]
- Neuronal cell protection assay: PC12 cells were subjected to OGD (4 hours) followed by reoxygenation (24 hours) with Tanshinone IIA (Dan Shen ketone) (0.5–4 μM). CCK-8 assay measured cell viability. DCFH-DA staining detected ROS production. Western blot analyzed p-ERK1/2 and p-JNK [3]

Male ICR mice (25–30 g)[4]
10 or 20 mg/kg
P.o.

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Lung cancer xenograft mouse model: 6–8-week-old BALB/c nude mice were subcutaneously injected with A549 cells (2×106 cells/mouse) into the right flank. When tumors reached ~100 mm³, mice were randomly divided into control and treatment groups. Tanshinone IIA (Dan Shen ketone) was dissolved in DMSO/normal saline (final DMSO ≤5%) and administered intraperitoneally at 10 or 20 mg/kg every 3 days for 4 weeks. Tumor volume (measured every 3 days) and weight (end of treatment) were recorded. Tumor tissues were analyzed for microvessel density and p-VEGFR2 expression [2]
- LPS-induced neuroinflammation mouse model: Male C57BL/6 mice were intraperitoneally injected with LPS (5 mg/kg) to induce neuroinflammation. Tanshinone IIA (Dan Shen ketone) was dissolved in normal saline and administered intraperitoneally at 5 or 10 mg/kg/day for 7 days (starting 1 day before LPS injection). Mice were sacrificed, cerebral cortex tissues collected for cytokine detection (TNF-α, IL-6) and microglia activation analysis [4]
- MCAO-induced cerebral ischemia rat model: Male Sprague-Dawley rats were subjected to MCAO for 2 hours. Tanshinone IIA (Dan Shen ketone) was dissolved in normal saline and intravenously injected at 5 mg/kg 2 hours after ischemia. Neurological deficit scores were evaluated 24 hours post-ischemia. Rats were sacrificed, brain tissues collected for infarct volume measurement (TTC staining) and antioxidant index detection (SOD, MDA) [3]

In vitro toxicity: Tanshinone IIA (0.5–20 μM) showed extremely low cytotoxicity to normal human hepatocytes (LO2) and primary astrocytes, with cell viability remaining above 85% at all tested concentrations [1,3]. In vivo toxicity: Intraperitoneal or intravenous injection of tanshinone IIA (5–20 mg/kg) did not significantly alter body weight, food intake, or obvious toxic symptoms (such as somnolence and diarrhea) in mice/rats. Serum ALT, AST, creatinine, and blood urea nitrogen levels remained within the normal range [2,3,4].

[1]. Eur J Pharmacol . 2007 Jul 30;568(1-3):213-21.

[2]. Int J Cancer . 2005 Sep 20;116(5):799-807.

[3]. Brain Res Bull . 2012 Sep 1;88(6):581-8.

[3]. 2014,5, 1528-1532

1,6,6-Trimethyl-8,9-dihydro-7H-naphtho[1,2-g]benzofuran-10,11-dione is an abiran diterpenoid compound. Tanshinone IIA has been reported in Salvia miltiorrhiza, Salvia miltiorrhiza var. mucilaginosa, and other organisms with relevant data. See also: Salvia miltiorrhiza root (partial). Mechanism of Action: Doxorubicin was one of the earliest anthracycline antibiotics and remains one of the most effective anticancer drugs. However, the clinical application of doxorubicin is greatly limited by its severe cardiac adverse reactions, which may ultimately lead to cardiomyopathy and heart failure. Tanshinone IIA is the main active ingredient of Salvia miltiorrhiza (a traditional Chinese medicine used to treat cardiovascular diseases). This study aimed to evaluate the protective effect of tanshinone IIA against doxorubicin-induced cardiomyocyte apoptosis and to explore its intracellular mechanism. Primary cultured neonatal rat cardiomyocytes were treated with solvent, doxorubicin (1 μM), tanshinone IIA (0.1, 0.3, 1, and 3 μM), or a combination of tanshinone IIA and doxorubicin. The study found that tanshinone IIA (1 and 3 μM) inhibited doxorubicin-induced reactive oxygen species (ROS) generation, reduced the levels of cleavable caspase-3 and cytochrome c, and increased Bcl-x(L) expression, thereby protecting cardiomyocytes from doxorubicin-induced apoptosis. Furthermore, tanshinone IIA treatment enhanced Akt phosphorylation in cardiomyocytes. Transfection with Watermanin (100 nM), LY294002 (10 nM), and Akt siRNA significantly reduced the protective effect of tanshinone IIA. These results suggest that tanshinone IIA partially protects cardiomyocytes from doxorubicin-induced apoptosis through the Akt signaling pathway, which may contribute to protecting the heart from the severe toxic effects of doxorubicin. Tanshinone IIA (tanshinone) is a lipophilic diterpenoid quinone isolated from the root of the traditional Chinese medicine Salvia miltiorrhiza [1,2,3,4]. Its biological mechanisms include: 1) inhibiting tumor growth and angiogenesis by targeting the VEGFR2-AKT/ERK pathway; 2) inducing apoptosis and cell cycle arrest (G0/G1 phase) in cancer cells; 3) alleviating inflammation by inhibiting the NF-κB signaling pathway; and 4) protecting neurons from ischemia/reperfusion injury by regulating the MAPK pathway and reducing oxidative stress [1,2,3,4]. It has shown potential therapeutic value in various cancers (hepatocellular carcinoma, lung cancer, colon cancer), neuroinflammatory diseases, and cerebral ischemia [1,2,3,4]. As a natural product, it exhibits good biocompatibility and low toxicity to normal cells and tissues at therapeutic concentrations [1,3,4].

C19H18O3

294.3444

294.125

568-72-9

164676

Pink to red solid

1.2±0.1 g/cm3

480.7±44.0 °C at 760 mmHg

205-207ºC

236.4±21.1 °C

0.0±1.2 mmHg at 25°C

1.588

5.47

0

3

0

22

509

0

O1C([H])=C(C([H])([H])[H])C2C(C(C3=C(C1=2)C([H])=C([H])C1=C3C([H])([H])C([H])([H])C([H])([H])C1(C([H])([H])[H])C([H])([H])[H])=O)=O

HYXITZLLTYIPOF-UHFFFAOYSA-N

InChI=1S/C19H18O3/c1-10-9-22-18-12-6-7-13-11(5-4-8-19(13,2)3)15(12)17(21)16(20)14(10)18/h6-7,9H,4-5,8H2,1-3H3

1,6,6-trimethyl-8,9-dihydro-7H-naphtho[1,2-g][1]benzofuran-10,11-dione

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product is not stable in solution, please use freshly prepared working solution for optimal results.

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

5% DMSO+30% PEG 300+2% Tween 80+ddH2O: 0.2mg/mL (Please use freshly prepared in vivo formulations for optimal results.)