Glucocorticoid Receptor (GR) [1]
Estrogen Receptor (ER) [1]
Liver X Receptor α (LXRα) [2]
Stearoyl-CoA Desaturase 1 (SCD1) [3]
PI3K, AKT, mTOR [4]

(20S)-Protopanaxatriol acts on human umbilical vein endothelial cells (HUVEC) via the glucocorticoid receptor (GR) and estrogen receptor (ER). With an EC50 of 482 nM, (20S)-Protopanaxatriol (PPT) raises [Ca2+]i in HUVEC. NO production is increased by ((20S)-Protopanaxatriol at 1 µM via ERβ [1]. (20S)-Protopanaxatriol inhibits SREBP-1c and its promoter's autonomous transactivation of Gal4-LXRα LBD. transcription dependent on T0901317. (20S)-Protopanaxatriol (10 μg/mL) inhibits RNA polymerase II's recruitment to the SREBP-1c LXRE region. Additionally, T0901317-dependent transcription of LXRα target genes involved in adipogenesis is inhibited by (20S)-Protopanaxatriol. LXRα target gene ABCA1's transcription was unaffected by transcription, although T0901317-induced cellular triglyceride (TG) accumulation in primary hepatocytes was decreased [2]. g-PPT (100 nM, 1 μM, 10 μM, 20 μM; 48 hours) decreased the expression of SCD1 in HCC827GR and H1975 cell lines [3].

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- (20S)-Protopanaxatriol specifically bound to GR and ER in human umbilical vein endothelial cells (HUVECs), promoting GR nuclear translocation and enhancing GR-mediated transcriptional activity. It also slightly activated ER-related gene expression (detected by Western blot and reporter gene assay) [1]
- In human hepatocellular carcinoma cells (HepG2), (20S)-Protopanaxatriol (10, 20, 40 μM) dose-dependently inhibited LXRα-mediated expression of lipogenic genes (ABCA1, SREBP-1c, FASN) at both mRNA and protein levels (qPCR, Western blot), and reduced intracellular lipid accumulation (Oil Red O staining) [2]
- In EGFR-TKI-resistant non-small cell lung cancer (NSCLC) cells (PC9/GR, H1975/GR), (20S)-Protopanaxatriol (5, 10, 20 μM) dose-dependently downregulated SCD1 protein expression, decreased the production of unsaturated fatty acids (oleic acid, palmitoleic acid), and inhibited lipid accumulation (HPLC, Oil Red O staining). When combined with EGFR-TKI (gefitinib), it significantly enhanced gefitinib's antiproliferative activity, reducing the IC50 from >10 μM (gefitinib alone) to 2.3 ± 0.3 μM (PC9/GR) and 3.1 ± 0.5 μM (H1975/GR), and induced cell apoptosis (Annexin V-FITC/PI staining) [3]
- In human breast cancer cells (MCF-7), (20S)-Protopanaxatriol (20, 40, 80 μM) dose-dependently inhibited cell proliferation (MTT assay) with an IC50 of 45.6 ± 3.2 μM. It induced apoptosis, increasing the apoptotic rate from 4.2% (control) to 38.5% (80 μM) (Annexin V-FITC/PI staining). Western blot showed downregulated phosphorylated PI3K, AKT, mTOR, and anti-apoptotic protein Bcl-2, as well as upregulated pro-apoptotic protein Bax and cleaved caspase-3 [4]

(20S)-Protopanaxatriol (10 mg/kg; intraperitoneally; once daily for four weeks) and Gefitinib work together to suppress xenograft growth [3]. (20S)-Protopanaxatriol suppresses the development of MCF-7 breast cancer cells in a nude mouse xenograft experiment (50–100 mg/kg; oral; 25 days; female BALB/c nude mice harboring breast cancer MCF-7 cells) [4].

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- In nude mice bearing PC9/GR cell-derived xenografts, when tumors reached ~100 mm³, mice were randomly divided into control, gefitinib (50 mg/kg/day, oral), (20S)-Protopanaxatriol (20 mg/kg/day, intraperitoneal injection), and combination (gefitinib + (20S)-Protopanaxatriol) groups, with treatment lasting 4 weeks. The combination group showed a 62.3% reduction in tumor volume and 58.7% reduction in tumor weight compared with the gefitinib-alone group (measured by caliper and weighing). Western blot of tumor tissues showed decreased SCD1 expression and increased Bax/cleaved caspase-3 expression. Immunohistochemistry revealed a reduced proportion of Ki-67-positive proliferating cells [3]

- GR binding assay: Recombinant GR ligand-binding domain was incubated with (20S)-Protopanaxatriol (0.1-100 μM) and a fluorescent GR ligand. Fluorescence polarization was measured to assess binding affinity [1]
- ER binding assay: Recombinant ER ligand-binding domain was incubated with (20S)-Protopanaxatriol (0.1-100 μM) and a fluorescent ER ligand. Binding rate was quantified by fluorescence polarization [1]
- LXRα transcriptional activity assay: HepG2 cells transfected with LXRα reporter gene plasmid were treated with (20S)-Protopanaxatriol (10-40 μM) for 24 hours. Luciferase activity was detected to evaluate the inhibition of LXRα-mediated transcription [2]
- SCD1 activity assay: Purified SCD1 protein was incubated with (20S)-Protopanaxatriol (1-20 μM) and stearoyl-CoA substrate. The production of unsaturated fatty acids was detected to quantify SCD1 activity inhibition rate [3]

- Endothelial cell receptor activity assay: HUVECs were seeded and treated with (20S)-Protopanaxatriol (10, 20, 40 μM) for 24 hours. GR nuclear translocation and ER downstream gene expression were detected by Western blot. For reporter gene assay, HUVECs transfected with GR or ER reporter plasmid were treated with the compound, and luciferase activity was measured [1]
- Hepatocyte lipogenesis assay: HepG2 cells were seeded and treated with (20S)-Protopanaxatriol (10-40 μM) for 48 hours. Lipid accumulation was observed by Oil Red O staining; mRNA expression of ABCA1, SREBP-1c, FASN was detected by qPCR; corresponding protein levels by Western blot [2]
- NSCLC drug resistance reversal assay: PC9/GR and H1975/GR cells were seeded and treated with (20S)-Protopanaxatriol (5-20 μM) alone or combined with gefitinib (1-10 μM) for 72 hours. Cell viability was detected by MTT assay; apoptosis by Annexin V-FITC/PI staining; fatty acid composition by HPLC; SCD1 and apoptosis-related proteins by Western blot [3]
- Breast cancer cell apoptosis assay: MCF-7 cells were seeded and treated with (20S)-Protopanaxatriol (20-80 μM) for 48 hours. Cell viability was detected by MTT assay; apoptosis by Annexin V-FITC/PI staining; PI3K/AKT/mTOR pathway and apoptosis-related proteins by Western blot [4]

Animal/Disease Models: H1975 mouse xenograft tumor model
Doses: 10 mg/kg
Route of Administration: intraperitoneal (ip) injection;
Experimental Results: Compared with gefitinib or g-PPT treatment alone, g-PPT and gefitinib (50 mg/kg kg/day) combined treatment Dramatically diminished p-EGFR and KI67 expression and increased c-Caspase3 expression.

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- Xenograft model of EGFR-TKI-resistant NSCLC: BALB/c nude mice were subcutaneously injected with PC9/GR cells (5×10⁶ cells/mouse). When tumors reached ~100 mm³, mice were randomly assigned to four groups. The gefitinib group received oral gefitinib (50 mg/kg/day), the (20S)-Protopanaxatriol group received intraperitoneal injection (20 mg/kg/day), the combination group received both, and the control group received vehicle. Treatment lasted 4 weeks. Tumor volume was measured every 3 days (volume = length × width² / 2). At the end of treatment, mice were sacrificed, tumors were weighed, and tumor tissues were collected for Western blot and immunohistochemistry [3]

[1]. Protopanaxadiol and protopanaxatriol bind to glucocorticoid and oestrogen receptors in endothelial cells. Br J Pharmacol. 2009 Feb;156(4):626-37.

[2]. 20(S)-protopanaxatriol inhibits liver X receptor α-mediated expression of lipogenic genes in hepatocytes. J Pharmacol Sci. 2015 Jun;128(2):71-7.

[3]. Co-administration of 20(S)-protopanaxatriol (g-PPT) and EGFR-TKI overcomes EGFR-TKI resistance by decreasing SCD1 induced lipid accumulation in non-small cell lung cancer. J Exp Clin Cancer Res. 2019;38(1):129. Published 2019 Mar 15.

[4]. 20(S)-Protopanaxadiol-Induced Apoptosis in MCF-7 Breast Cancer Cell Line through the Inhibition of PI3K/AKT/mTOR Signaling Pathway. Int J Mol Sci. 2018;19(4):1053. Published 2018 Apr 2.

Protopanaxatriol is a tetracyclic triterpenoid saponin (isolated from ginseng and Panax notoginseng). It is a derivative of dammarane, with hydroxyl groups substituted at positions 3β, 6α, 12β, and 20 (pro-S position), and a double bond introduced at positions 24-25. It is a metabolite. It is a tetracyclic triterpenoid compound, saponin, 3β-hydroxysteroid, 12β-hydroxysteroid, 6α-hydroxysteroid, and 3β-hydroxy-4,4-dimethylsteroid. It is derived from the hydride of dammarane. Protopanaxatriol has been reported to be present in Gynostemma pentaphyllum, ginseng, and other organisms with relevant data.

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- (20S)-Protopanaxatriol is a natural triterpenoid compound isolated from plants of the Araliaceae family (such as ginseng)[1][2][3][4]
- Its core mechanisms include: binding to glucocorticoid receptor (GR) and estrogen receptor (ER) to regulate endothelial cell function[1]; inhibiting LXRα-mediated adipogenesis gene expression to reduce lipid accumulation[2]; downregulating SCD1 to reverse lipid metabolism reprogramming in EGFR-TKI resistant non-small cell lung cancer (NSCLC)[3]; and inhibiting PI3K/AKT/mTOR signaling pathway to induce apoptosis in breast cancer cells[4]
- It has shown potential therapeutic value in studies of metabolic-related diseases, EGFR-TKI resistant non-small cell lung cancer and breast cancer[2][3][4].

C30H52O4

476.7315

476.386

34080-08-5

11468733

White to off-white solid powder

1.1±0.1 g/cm3

588.8±50.0 °C at 760 mmHg

242.9±24.7 °C

0.0±3.7 mmHg at 25°C

1.541

5.41

4

4

4

34

817

11

CC(=CCC[C@@](C)([C@H]1CC[C@@]2([C@@H]1[C@@H](C[C@H]3[C@]2(C[C@@H]([C@@H]4[C@@]3(CC[C@@H](C4(C)C)O)C)O)C)O)C)O)C

SHCBCKBYTHZQGZ-CJPZEJHVSA-N

InChI=1S/C30H52O4/c1-18(2)10-9-13-30(8,34)19-11-15-28(6)24(19)20(31)16-22-27(5)14-12-23(33)26(3,4)25(27)21(32)17-29(22,28)7/h10,19-25,31-34H,9,11-17H2,1-8H3/t19-,20+,21-,22+,23-,24-,25-,27+,28+,29+,30-/m0/s1

(3S,5R,6S,8R,9R,10R,12R,13R,14R,17S)-17-[(2S)-2-hydroxy-6-methylhept-5-en-2-yl]-4,4,8,10,14-pentamethyl-2,3,5,6,7,9,11,12,13,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene-3,6,12-triol

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)

DMSO : ~100 mg/mL (~209.76 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.36 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 20.8 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.36 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 20.8 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.36 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)