Curzerene (0-100 µM; 24-72 hours) showed dose- and time-dependently increasing cytostatic effects against SPC A1 cells with IC50s of 403.8 µM, 154.8 µM, and 47.01 µM at 24, 48, and 72 hours, respectively [1]. In comparison to controls, Curzerene (0-100 µM; 48 hours) causes greater proportions of necrotic and apoptotic cells in SPC-A1 cells [1]. The percentage of cells arrested in the G2/M phase rose from 9.26% of cells in the control group to 17.57% of cells treated with the highest dose, according to research using curzerene (0-100 µM; 48 hours) [1]. In SPC A1 cells, curzerene (6.25-100 µM; 48 hours) decreases GSTA1 mRNA expression [1]. In SPC A1 cells, curzerene (6.25-100 µM; 48 hours) can lower GSTA1 protein expression [1].

Curzerene (0-100 µM; 24-72 hours) showed dose- and time-dependently increasing cytostatic effects against SPC A1 cells with IC50s of 403.8 µM, 154.8 µM, and 47.01 µM at 24, 48, and 72 hours, respectively [1]. In comparison to controls, Curzerene (0-100 µM; 48 hours) causes greater proportions of necrotic and apoptotic cells in SPC-A1 cells [1]. The percentage of cells arrested in the G2/M phase rose from 9.26% of cells in the control group to 17.57% of cells treated with the highest dose, according to research using curzerene (0-100 µM; 48 hours) [1]. In SPC A1 cells, curzerene (6.25-100 µM; 48 hours) decreases GSTA1 mRNA expression [1]. In SPC A1 cells, curzerene (6.25-100 µM; 48 hours) can lower GSTA1 protein expression [1].

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Antiproliferative Activity: Curzerene exhibited antiproliferative effects on SPC-A1 human lung adenocarcinoma cells in a time- and dose-dependent manner, as determined by MTT assay. The IC₅₀ values were 403.8 µM at 24 hours, 154.8 µM at 48 hours, and 47.01 µM at 72 hours. At 25 µM, its inhibition rate was similar to the positive control (100 µM β-elemene). [1]
Apoptosis Induction: Flow cytometry analysis using Annexin V-FITC/PI staining showed that treatment with curzerene (6.25, 12.5, 25, 50, 100 µM) for 48 hours induced apoptosis in SPC-A1 cells in a dose-dependent manner. Hoechst 33258 staining revealed nuclear condensation, a morphological hallmark of apoptosis, in treated cells. [1]
Cell Cycle Arrest: Flow cytometric cell cycle analysis (PI staining) demonstrated that curzerene treatment for 48 hours caused a dose-dependent arrest of SPC-A1 cells at the G2/M phase. The percentage of cells in G2/M increased from 9.26% in the control to 17.57% at the highest dose (100 µM). [1]
Downregulation of GSTA1: Western blot and quantitative RT-PCR analyses showed that treatment with curzerene (6.25, 25, 100 µM) for 48 hours significantly downregulated both the protein and mRNA expression levels of GSTA1 in SPC-A1 cells compared to the control. [1]

Antitumor Efficacy: In SPC-A1 cell-bearing BALB/c nude mice, daily intraperitoneal administration of curzerene for 12 days significantly inhibited tumor growth in a dose-dependent manner. Tumor growth inhibition rates were 19.71%, 32.58%, and 58.94% for the low (15 mg/kg), medium (45 mg/kg), and high (135 mg/kg) dose groups, respectively. The high-dose group showed superior efficacy compared to the positive control (β-elemene, 135 mg/kg). [1]
Body Weight and Organ Coefficients: During the 12-day treatment period, all mice, including those treated with curzerene, gained weight. The liver organ coefficients in the medium- and high-dose curzerene groups were higher than in the model control group, but other organ coefficients (heart, spleen, lung, kidney) were not significantly different. This suggests limited systemic toxicity. [1]

Cell Viability Assay[1]
Cell Types: SPC-A1 Cell
Tested Concentrations: 0 µM, 6.25 µM, 12.5 µM, 25 µM, 50 µM, 100 µM
Incubation Duration: 24 hrs (hours), 48 hrs (hours), 72 hrs (hours)
Experimental Results: Inhibition of non-cell growth In vitro experiments on lung cancer SPC A1 cells.

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Apoptosis analysis[1]
Cell Types: SPC-A1 Cell
Tested Concentrations: 0 µM, 6.25 µM, 12.5 µM, 25 µM, 50 µM, 100 µM
Incubation Duration: 48 hrs (hours)
Experimental Results: Induction of apoptosis in a dose-dependent manner.

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Apoptosis analysis[1]
Cell Types: SPC-A1 Cell
Tested Concentrations: 0 µM, 6.25 µM, 12.5 µM, 25 µM, 50 µM, 100 µM
Incubation Duration: 48 hrs (hours)
Experimental Results: Induction of G2/M cell cycle in SPC A1 cells Stagnation.

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RT-PCR[1]
Cell Types: SPC-A1 Cell
Tested Concentrations: 6.25 µM, 25 µM, 100 µM
Incubation Duration: 48 hrs (hours)
Experimental Results: diminished GSTA1 mRNA expression.

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MTT Assay for Cell Viability: SPC-A1 cells were seeded in 96-well plates. After 24 hours of attachment, cells were treated with various concentrations (0, 6.25, 12.5, 25, 50, 100 µM) of curzerene or positive control (100 µM β-elemene) for 24, 48, or 72 hours. Curzerene was dissolved in absolute ethanol, and the final treatment medium contained 0.1% ethanol. After incubation, MTT solution was added to each well. Following a 4-hour incubation, the supernatant was discarded, DMSO was added, and the absorbance at 570 nm was measured with a microplate reader to calculate cell viability. [1]
Hoechst 33258 Staining for Apoptotic Morphology: SPC-A1 cells seeded in 24-well plates were treated with curzerene (0–100 µM) or positive control for 48 hours. The supernatant was discarded, cells were fixed with 4% paraformaldehyde, washed, stained with Hoechst 33258 dye, washed again, and observed under a fluorescence microscope for nuclear condensation. [1]
Annexin V-FITC/PI Staining for Apoptosis by Flow Cytometry: SPC-A1 cells in the logarithmic growth phase were seeded in 6-well plates and treated with curzerene (0–100 µM) or positive control for 48 hours. Cells were collected, centrifuged, resuspended in binding buffer, stained with Annexin V-FITC and PI in the dark, and analyzed by flow cytometry to determine the apoptosis rate. [1]
PI Staining for Cell Cycle Analysis by Flow Cytometry: SPC-A1 cells were seeded in 6-well plates and treated with curzerene (0–100 µM) or positive control for 48 hours. Cells were harvested, washed, fixed in 70% ethanol, treated with RNase, stained with PI, and analyzed by flow cytometry. The percentage of cells in G0/G1, S, and G2/M phases was determined using appropriate software. [1]
RT-PCR for GSTA1 mRNA Expression: SPC-A1 cells were seeded in 6-well plates and divided into six groups: blank control (serum-free medium), solvent control (0.1% ethanol in serum), positive control (100 µM β-elemene), and three curzerene groups (6.25, 25, 100 µM). After 48 hours of treatment, total RNA was extracted, cDNA was synthesized, and real-time PCR was performed using specific primers for GSTA1 and GAPDH (for normalization). [1]
Western Blot for GSTA1 Protein Expression: SPC-A1 cells were treated and grouped as for the RT-PCR experiment. After 48 hours, cells were collected and protein was extracted. Protein samples were separated by SDS-PAGE, transferred to a membrane, blocked, and incubated with primary antibody against GSTA1 overnight, followed by incubation with a secondary antibody. Protein bands were visualized and quantified using imaging software. [1]

In Vivo Antitumor Efficacy Study in Nude Mice: BALB/c nude mice were subcutaneously injected in the right flank with SPC-A1 cells. When tumors reached approximately 4–5 mm in diameter, mice were randomly allocated into groups. Curzerene was dissolved in ethanol and diluted in saline to a final concentration containing 1% ethanol. The drug was administered via intraperitoneal injection at doses of 15, 45, or 135 mg/kg, once daily for 12 consecutive days. The injection volume was 0.1 mL per 10 g of body weight. Mouse body weights were measured daily, and tumor volumes (calculated as 0.5 × length × width²) were measured every two days. After 12 days, mice were sacrificed under anesthesia. Tumors and organs (heart, liver, spleen, lung, kidney) were harvested and weighed. [1]

In vivo toxicity analysis: In the SPC-A1 tumor-bearing nude mouse model, intraperitoneal injection of curzerene (15, 45, 135 mg/kg) for 12 consecutive days did not lead to animal mortality. All mice, including the treatment group, gained weight during the experiment. Although the liver organ coefficients were increased in the medium- and high-dose groups compared with the model control group, the coefficients of other major organs were not significantly affected. The authors believe that this indicates that curzerene has limited toxicity and side effects in vivo. [1]

[1]. Cytotoxic and Antitumor Effects of Curzerene from Curcuma longa. Planta Med. 2017 Jan;83(1-02):23-29.

Benzofuran, 6-vinyl-4,5,6,7-tetrahydro-3,6-dimethyl-5-isopropenyl-, trans-, has been reported in turmeric (Curcuma xanthorrhiza), Litsea cubeba (Lindera pulcherrima var. hemsleyana), and other organisms with relevant data. Curcumin is a sesquiterpene isolated from the rhizome of turmeric (Curcuma longa) and is the main component of turmeric oil. This study proposes that curcumin may be a potential candidate drug for treating lung adenocarcinoma. Its anticancer mechanism in SPC-A1 cells includes inducing cell cycle arrest in the G2/M phase, promoting apoptosis, and downregulating the drug resistance-associated detoxification enzyme GSTA1. The authors suggest that future studies should combine curcumin with standard chemotherapy drugs such as carboplatin or cisplatin for the treatment of non-small cell lung cancer. [1]

C15H20O

216.3187

216.151

17910-09-7

12305301

Colorless to light yellow liquid

1.0±0.1 g/cm3

282.8±40.0 °C at 760 mmHg

117.5±14.2 °C

0.0±0.6 mmHg at 25°C

1.539

5.78

0

1

2

16

307

2

O1C([H])=C(C([H])([H])[H])C2=C1C([H])([H])C(C([H])=C([H])[H])(C([H])([H])[H])C([H])(C(=C([H])[H])C([H])([H])[H])C2([H])[H]

HICAMHOOTMOHPA-HIFRSBDPSA-N

InChI=1S/C15H20O/c1-6-15(5)8-14-12(11(4)9-16-14)7-13(15)10(2)3/h6,9,13H,1-2,7-8H2,3-5H3/t13-,15+/m1/s1

(5R,6R)-6-ethenyl-3,6-dimethyl-5-prop-1-en-2-yl-5,7-dihydro-4H-1-benzofuran

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.  (3). 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)

DMSO : ~250 mg/mL (~1155.70 mM)

Solubility in Formulation 1: ≥ 6.25 mg/mL (28.89 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 62.5 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: 6.25 mg/mL (28.89 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 62.5 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: ≥ 6.25 mg/mL (28.89 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 62.5 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly.

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