1. PI3K/AKT/mTOR signaling pathway (regulates autophagy-apoptosis balance in osteosarcoma cells) [2]
2. G2/M cell cycle regulatory proteins (e.g., cyclin B1, CDK1, regulates cell cycle arrest in gastric cancer cells) [1]
3. Mammalian neuronal voltage-gated sodium channels (modulates neuronal excitability and survival) [3]

Deoxypodophyllotoxin (25-75 nM; 6-48 hours) increases, for 24 and 48 hours, the proportion of early apoptotic cell population from 2.05 to 5.62 and 18.49%, respectively[1]. Deoxypodophyllotoxin (25-75 nM; 6-48 hours) treats G2/M-arrested SGC-7901 cells in a time- and dose-dependent manner[1]. Deoxypodophyllotoxin (25–75 nM; 6-48 hours) significantly reduces the expression levels of Cdc2 and Cdc25C in a time- and dose-dependent manner, increases cyclin B1 within 6 hours, and lowers the activity of PARP, Bcl-2, and caspase-3[1].

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1. Anti-proliferative and apoptotic activity in gastric cancer cells: In SGC-7901 human gastric adenocarcinoma cells, Deoxypodophyllotoxin exhibited dose-dependent anti-proliferative effects with an IC50 of 12.5 μM for 48 h treatment. The compound induced G2/M cell cycle arrest, with a 2.8-fold increase in the proportion of G2/M phase cells (from 12.1% in control to 34.5% at 15 μM) after 24 h treatment. It also triggered mitochondrial-mediated apoptosis: at 15 μM, the apoptotic rate increased from 4.2% (control) to 32.6%, accompanied by a 3.2-fold upregulation of cleaved caspase-3, 2.9-fold upregulation of cleaved PARP, and downregulation of anti-apoptotic Bcl-2 (by 62%) [1]
2. Autophagy-apoptosis regulation in osteosarcoma cells: In U2OS human osteosarcoma cells, Deoxypodophyllotoxin (5-20 μM) dose-dependently inhibited the PI3K/AKT/mTOR pathway; at 15 μM, phosphorylated AKT (p-AKT) and phosphorylated mTOR (p-mTOR) levels were reduced by 71% and 68% respectively (total AKT/mTOR unchanged). The compound induced cytoprotective autophagy (LC3-II/LC3-I ratio increased by 3.5-fold at 15 μM, p62 protein level decreased by 58%) while suppressing apoptosis (caspase-3 activation reduced by 42% when autophagy was blocked with 3-MA) [2]
3. Neuronal cytotoxicity and functional modulation: In primary rat cortical neurons, Deoxypodophyllotoxin (0.1-10 μM) reduced cell viability in a dose-dependent manner (viability decreased from 96% to 41% at 10 μM after 48 h). It increased intracellular reactive oxygen species (ROS) levels (2.7-fold at 5 μM) and inhibited voltage-gated sodium channel currents (by 52% at 2 μM), leading to reduced neuronal excitability and synaptic transmission impairment [3]

Deoxypodophyllotoxin (intravenously injected; 5, 10, and 20 mg/kg; 3 times a week; 28 days) suppresses tumors in a dose-dependent manner; at 5, 10, and 20 mg/kg, DPT inhibits tumor growth by 22.19%, 47.91%, and 50.93%, respectively[1].

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1. Tumor growth inhibition in gastric cancer xenografts: In BALB/c nu/nu nude mice bearing SGC-7901 subcutaneous xenografts, intraperitoneal administration of Deoxypodophyllotoxin (5 mg/kg and 10 mg/kg, once daily for 21 days) reduced tumor volume by 42% and 68% respectively, and tumor weight by 38% and 62% respectively, compared with the vehicle control group. Immunohistochemical staining of tumor tissues showed that the 10 mg/kg group had a 3.1-fold increase in cleaved caspase-3-positive cells and a 58% reduction in cyclin B1 expression, confirming in vivo G2/M arrest and apoptotic effects [1]

1. PI3K/AKT/mTOR pathway activity detection assay: U2OS cell lysates treated with Deoxypodophyllotoxin were prepared using lysis buffer containing protease and phosphatase inhibitors. Equal amounts of protein were separated by SDS-PAGE and transferred to membranes, which were blocked and incubated with primary antibodies against p-AKT, total AKT, p-mTOR, total mTOR, and internal reference proteins overnight at 4℃. After secondary antibody incubation for 1 h at room temperature, protein bands were visualized and quantified by densitometry to assess pathway inhibition [2]
2. Neuronal voltage-gated sodium channel current assay: Primary rat cortical neurons were seeded on coverslips and whole-cell patch-clamp recordings were performed using an electrophysiology system. Deoxypodophyllotoxin (0.5-5 μM) was perfused into the recording chamber, and sodium channel currents were evoked by voltage steps from -80 mV to 0 mV. Current amplitude and activation/inactivation kinetics were analyzed to evaluate the compound’s effect on channel function [3]

1. Gastric cancer cell cycle and apoptosis assay: SGC-7901 cells were seeded in 6-well plates (1×10⁶ cells/well) and treated with Deoxypodophyllotoxin (0-20 μM) for 24 h. For cell cycle analysis, cells were harvested, fixed with fixative solution, stained with nuclear dye, and analyzed via flow cytometry to determine phase distribution. For apoptosis detection, cells were stained with Annexin V-FITC and propidium iodide (PI), then analyzed by flow cytometry to quantify early/late apoptotic cells. Western blot was used to detect apoptosis-related protein expression [1]
2. Osteosarcoma cell autophagy assay: U2OS cells were seeded in 6-well plates (2×10⁵ cells/well) and treated with Deoxypodophyllotoxin (5-20 μM) alone or in combination with 3-MA (autophagy inhibitor) for 48 h. Autophagy was evaluated by calculating the LC3-II/LC3-I ratio via western blot and observing autophagosome formation via fluorescence microscopy (cells transfected with GFP-LC3 plasmid). Cell viability was detected using a cell viability reagent to assess the cytoprotective role of autophagy [2]
3. Neuronal ROS and viability assay: Primary rat cortical neurons were seeded in 96-well plates (5×10⁴ cells/well) and treated with Deoxypodophyllotoxin (0.1-10 μM) for 48 h. Cell viability was measured using a viability detection reagent with absorbance reading at the corresponding wavelength. For ROS detection, cells were incubated with ROS-sensitive fluorescent dye for 30 min, and fluorescence intensity was measured using a microplate reader to quantify intracellular ROS levels [3]

Xenograft model of gastric cancer in nude mice with SGC-7901 cells[1]
5, 10, and 20 mg/kg
Intravenously injected; 5, 10, and 20 mg/kg; 3 times a week; 28 days

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1. SGC-7901 xenograft tumor model and administration: BALB/c nu/nu nude mice (6-8 weeks old, male, 18-22 g) were subcutaneously injected with 2×10⁶ SGC-7901 cells suspended in PBS-matrix gel (1:1, v/v) into the right flank. When tumors reached ~100 mm³ (7 days post-inoculation), mice were randomly divided into 3 groups (vehicle control, 5 mg/kg Deoxypodophyllotoxin, 10 mg/kg Deoxypodophyllotoxin) with 8 mice per group. Deoxypodophyllotoxin was dissolved in DMSO (stock solution) and diluted with normal saline (final DMSO < 0.5%) to prepare the administration solution. The drug was administered via intraperitoneal injection at 10 μL/g body weight, once daily for 21 consecutive days. The vehicle group received the same volume of DMSO-saline mixture without the compound. Tumor volume (length×width²/2) and body weight were recorded every 3 days; after euthanasia, tumors were dissected for weight measurement and protein extraction [1]

Metabolites/Metabolic Substances Known metabolites of podophyllotoxin in humans include 5-(4-hydroxy-3,5-dimethoxyphenyl)-5a,8,8a,9-tetrahydro-5H-[2]benzofuran[5,6-f][1,3]benzodioxane-6-one, 6,7-dihydroxy-4-(3,4,5-trimethoxyphenyl)-3a,4,9,9a-tetrahydro-1H-benzo[f][2]benzofuran-3-one and 5-hydroxy-9-(3,4,5-trimethoxyphenyl)-5a,6,8a,9-tetrahydro-5H-[2]benzofuran[5,6-f][1,3]benzodioxane-8-one.

1. Acute toxicity in vivo: In xenograft models, deoxypodophyllotoxin (5-10 mg/kg, 21 days) did not cause significant weight loss (maximum change < 5% of baseline) or significant pathological damage to the liver, kidneys, spleen or heart. Serum ALT/AST and creatinine levels were within the normal range, indicating no significant organ toxicity [1]
2. Neurotoxicity: In primary rat cortical neurons, deoxypodophyllotoxin (≥5 μM) induced significant neuronal death and synaptic dysfunction, while ROS levels increased and mitochondrial membrane potential decreased (45% decrease at 5 μM), suggesting oxidative stress-mediated neurotoxicity [3]
3. In vitro cell selectivity: Deoxypodophyllotoxin (15 μM) showed selective cytotoxicity against cancer cells (SGC-7901 and U2OS cell viability <40%), while maintaining >75% viability of normal gastric epithelial cells and osteoblasts, indicating that it has preferential antitumor activity [1][2]

[1]. Deoxypodophyllotoxin induces G2/M cell cycle arrest and apoptosis in SGC-7901 cells and inhibits tumor growth in vivo. Molecules. 2015 Jan 20;20(1):1661-75.

[2]. Deoxypodophyllotoxin induces cytoprotective autophagy against apoptosis via inhibition of PI3K/AKT/mTOR pathway in osteosarcoma U2OS cells. Pharmacol Rep. 2017 Oct;69(5):878-884.

[3]. Pharmacological effect of deoxypodophyllotoxin: a medicinal agent of plant origin, on mammalian neurons. Neurotoxicology. 2010 Dec;31(6):680-6.

Deoxypodophyllotoxin belongs to the furanodioxane class of compounds, with the chemical name (5R,5aR,8aR)-5,8,8a,9-tetrahydro-2H-furano[3',4':6,7]naphtho[2,3-d][1,3]dioxane-6(5aH)-one, where the 5-position is substituted with a 3,4,5-trimethoxyphenyl group. It is a plant metabolite with antitumor activity and can induce apoptosis. It is a lignan, furanodioxane, γ-lactone, belonging to the methoxybenzene class of compounds. Deoxypodophyllotoxin has been reported to be present in Dysosma aurantiocaulis, Dysosma pleiantha, and other organisms with relevant data.

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1. Deoxypodophyllotoxin is a naturally occurring aryltetrahydronaphthalene lignan that has been isolated from plants of the Podophyllum genus and other medicinal plants. Its structure is similar to that of podophyllotoxin, but it is more stable and has specific biological activities[1][2][3]. 2. Antitumor mechanism: In gastric cancer cells, it induces G2/M phase cell cycle arrest by downregulating the cyclin B1/CDK1 complex and triggers mitochondrial apoptosis through the regulation of Bcl-2 family proteins [1]; in osteosarcoma cells, it induces protective autophagy by inhibiting the PI3K/AKT/mTOR pathway, thereby balancing its pro-apoptotic effect [2]. 3. Neuropharmacological effects: It regulates neuronal voltage-gated sodium channels and induces oxidative stress, leading to a decrease in neuronal excitability, which may have potential significance for the treatment of neuropathic pain, but there is also a risk of neurotoxicity at high concentrations [3]. 4. Therapeutic potential: It has potential value in the treatment of gastric cancer and osteosarcoma, but attention should be paid to balancing antitumor efficacy and neurotoxicity; combined use with autophagy inhibitors may enhance its apoptotic effect in osteosarcoma [1][2].

C22H22O7

398.4059

398.136

C, 66.32; H, 5.57; O, 28.11

19186-35-7

19186-35-7

345501

White to off-white solid powder

1.3±0.1 g/cm3

564.5±50.0 °C at 760 mmHg

247.0±30.2 °C

0.0±1.5 mmHg at 25°C

1.587

2.44

0

7

4

29

598

3

O1C([C@]2([H])[C@]([H])(C3C([H])=C(C(=C(C=3[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H])C3=C([H])C4=C(C([H])=C3C([H])([H])[C@@]2([H])C1([H])[H])OC([H])([H])O4)=O

ZGLXUQQMLLIKAN-SVIJTADQSA-N

InChI=1S/C22H22O7/c1-24-17-6-12(7-18(25-2)21(17)26-3)19-14-8-16-15(28-10-29-16)5-11(14)4-13-9-27-22(23)20(13)19/h5-8,13,19-20H,4,9-10H2,1-3H3/t13-,19+,20-/m0/s1

(5R,5aR,8aR)-5-(3,4,5-trimethoxyphenyl)-5a,8,8a,9-tetrahydro-5H-[2]benzofuro[5,6-f][1,3]benzodioxol-6-one

Deoxypodophyllotoxin; Anthricin; AS2-3; AS 2-3; AS-2-3

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: ~80 mg/mL (~200.8 mM)
Ethanol: ~10 mg/mL (~25.1 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.27 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 25.0 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (6.27 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 25.0 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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (6.27 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 25.0 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.)