Natural sesquiterpene lactone; antibacterial, anticancer and antifungal
NF-κB (p65) pathway. [1]
NF-κB (p65) pathway. [5]
Farnesyltransferase (FTase) with an IC50 of 64 µM. The study suggests that the antiproliferative effect in MDA-MB-231 cells is independent of FTase inhibition. [4]

Xanthatin is a naturally occurring sesquiterpene lactone,with significant antitumor activity against a variety of cancer cells; it exhibits significant antitumor effects through cell cycle arrest and apoptosis induction in A549 cells, these effects are associated with intrinsic apoptosis pathway and disrupted NF-κB signaling, suggests that it may have therapeutic potential against human non-small-cell lung cancer.[1]
Xanthatin has bactericidal and fungicidal activity against Colletotrichum gloesporoides, Trichothecium roseum, Bacillus cereus and Staphylococcus aureus.[2]
Xanthatin and the crude extracts of Xanthium strumarium have cytotoxic activity.[3]
(−)-Xanthatin is a highly effective inhibitor of MDA-MB-231 cell growth, inducing caspase-independent cell death, and that these effects were independent of FTase inhibition; GADD45γ was selectively induced by (−)-xanthatin and that GADD45γ-primed JNK and p38 signaling pathways are, at least in part, involved in mediating the growth inhibition and potential anticancer activities of this agent; GADD45γ is becoming increasingly recognized for its tumor suppressor function, suggests that the novel possibility that (−)-xanthatin may have therapeutic value as a selective inducer of GADD45γ in human cancer cells, in particular in FTI-resistant aggressive breast cancers.[4]
Xanthatin induces G2/M cell cycle arrest and apoptosis in human gastric carcinoma MKN-45 cells, it may have therapeutic potential against human gastric carcinoma.[5]
Xanthatin is a novel potent inhibitor of VEGFR2 signaling, can inhibit angiogenesis and tumor growth in breast cancer cells.[6]

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Xanthatin induced obvious dose-/time-dependent cytotoxicity against A549 NSCLC cells (IC50 values not specified for this cell line in this paper). It induced cell cycle arrest at the G2/M phase and promoted apoptosis. Mechanistically, it downregulated Chk1, Chk2, and phosphorylation of CDC2, leading to cell cycle arrest. It increased total p53 protein levels, decreased the Bcl-2/Bax ratio, and reduced expression of procaspase-9 and procaspase-3, triggering the intrinsic apoptosis pathway. Xanthatin also blocked phosphorylation of NF-κB (p65) and IκBα and inhibited TNFα-induced NF-κB (p65) translocation. [1]
Xanthatin showed in vitro cytotoxicity against murine lymphocytic leukemia P-388 and L-1210 cell lines with IC50 values of 0.018 µg/mL and 0.009 µg/mL, respectively. It also exhibited activity against the human bronchial epidermoid carcinoma NSCLC-N6 cell line with an IC50 of 3 µg/mL. [3]
Among six synthesized xanthanolides, (-)-Xanthatin was a highly effective inhibitor of MDA-MB-231 breast cancer cell proliferation (IC50 = 5.28 µM after 48h). It suppressed cell viability in a time- and dose-dependent manner. It induced morphological changes (rounding of cells) and, at higher concentrations (25 µM), induced LDH release. It had no effect on p21 gene expression but up-regulated stress-responsive genes IL-1β (3.0-fold) and HO-1 (5.3-fold). The antiproliferative effects were suppressed by antioxidants N-acetyl-L-cysteine and Vitamin C. (-)-Xanthatin selectively induced GADD45γ mRNA expression (22.2-fold). It down-regulated Cdc2 and cyclin B1 (3.0 and 3.3-fold respectively). It did not cause oligonucleosomal DNA fragmentation but generated high molecular weight DNA fragments, suggesting caspase-independent cell death. It had no effect on Topoisomerase I-mediated DNA relaxation. (-)-Xanthatin also suppressed the viability of MCF-7 breast cancer cells (IC50 = 5.05 µM). [4]
Xanthatin showed significant antiproliferative effects on MKN-45 gastric carcinoma cells with IC50 values of 18.6, 9.3, and 3.9 µM for 12, 24, and 48 h, respectively. It induced G2/M cell cycle arrest and apoptosis. It downregulated Chk1, Chk2, and p-CDC2. It increased p53 activation, decreased the Bcl-2/Bax ratio, and reduced procaspase-9 and procaspase-3. It also blocked phosphorylation of NF-κB (p65) and IκBα. [5]

Xanthatin was tested in vivo on murine ascite leukemia P-388. It showed weak activity. The T/C (ratio of mean survival time of test group vs. control) was 127% at 80 mg/kg/day and 134% at 75 mg/kg/day. The product is considered interesting if T/C is ≥125%. [3]

A relaxation assay of DNA Topoisomerase I (Topo I) was performed. Topo I and pBR322 DNA (supercoiled DNA) were used. The enzyme reaction was performed according to the manufacturer's protocol. The reaction products were analyzed by agarose gel electrophoresis. Supercoiled DNA was almost completely converted into relaxed DNA in the presence of 1 or 2 U Topo I. Under these conditions, no inhibitory effect of 10 µM (-)-Xanthatin on Topo I was detected (up to 25 µM). [4]

Xanthatin, a natural sesquiterpene lactone, has significant antitumor activity against a variety of cancer cells, yet little is known about its anticancer mechanism. In this study, we demonstrated that xanthatin had obvious dose-/time-dependent cytotoxicity against the human non-small-cell lung cancer (NSCLC) cell line A549. Flow cytometry analysis showed xanthatin induced cell cycle arrest at G2/M phase. Xanthatin also had pro-apoptotic effects on A549 cells as evidenced by Hoechst 33258 staining and annexin V-FITC staining. Mechanistic data revealed that xanthatin downregulated Chk1, Chk2, and phosphorylation of CDC2, which contributed to the cell cycle arrest. Xathatin also increased total p53 protein levels, decreased Bcl-2/Bax ratio and expression of the downstream factors procaspase-9 and procaspase-3, which triggered the intrinsic apoptosis pathway. Furthermore, xanthatin blocked phosphorylation of NF-κB (p65) and IκBa, which might also contribute to its pro-apoptotic effects on A549 cells. Xanthatin also inhibited TNFa induced NF-κB (p65) translocation. We conclude that xanthatin displays significant antitumor effects through cell cycle arrest and apoptosis induction in A549 cells. These effects were associated with intrinsic apoptosis pathway and disrupted NF-κB signaling. These results suggested that xanthatin may have therapeutic potential against NSCLC.[1]

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exo-Methylene lactone group-containing compounds, such as (--)-xanthatin, are present in a large variety of biologically active natural products, including extracts of Xanthium strumarium (Cocklebur). These substances are reported to possess diverse functional activities, exhibiting anti-inflammatory, antimalarial, and anticancer potential. In this study, we synthesized six structurally related xanthanolides containing exo-methylene lactone moieties, including (--)-xanthatin and (+)-8-epi-xanthatin, and examined the effects of these chemically defined substances on the highly aggressive and farnesyltransferase inhibitor (FTI)-resistant MDA-MB-231 cancer cell line. The results obtained demonstrate that (--)-xanthatin was a highly effective inhibitor of MDA-MB-231 cell growth, inducing caspase-independent cell death, and that these effects were independent of FTase inhibition. Further, our results show that among the GADD45 isoforms, GADD45γ was selectively induced by (--)-xanthatin and that GADD45γ-primed JNK and p38 signaling pathways are, at least in part, involved in mediating the growth inhibition and potential anticancer activities of this agent. Given that GADD45γ is becoming increasingly recognized for its tumor suppressor function, the results presented here suggest the novel possibility that (--)-xanthatin may have therapeutic value as a selective inducer of GADD45γ in human cancer cells, in particular in FTI-resistant aggressive breast cancers.[4]

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Cell viability was assessed using the MTS assay. Cells were seeded in 96-well plates and exposed to Xanthatin for indicated times. MTS/PMS solution was added and incubated, and the absorbance was measured at 490 nm. [1]
Cell cycle analysis was performed by flow cytometry. Cells were treated with Xanthatin, harvested, fixed with 70% alcohol, and stained with a DNA staining solution. The percentage of cells in each cell cycle phase was determined using a flow cytometer. [1]
Apoptosis was analyzed by Annexin-V/PI double staining. Cells were treated with Xanthatin, harvested, and stained with FITC-conjugated Annexin V and PI. Apoptosis was analyzed by fluorescence microscopy and flow cytometry. [1]
Hoechst 33258 staining was used to visualize nuclear morphology. Cells were fixed with 4% formaldehyde, stained with Hoechst 33258, and observed under a fluorescence microscope. [1]
Western blot analysis was performed. Whole cell proteins were extracted, separated by SDS-PAGE, transferred to PVDF membranes, and probed with specific primary antibodies followed by HRP-conjugated secondary antibodies. Proteins were visualized using enhanced chemiluminescence. [1]
NF-κB (p65) translocation was examined by immunocytochemistry. Cells were pretreated with Xanthatin and then stimulated with TNFα. Cells were fixed, permeabilized, and incubated with anti-p65 antibody, followed by a fluorescent secondary antibody. Cells were observed under a laser scanning confocal microscope. Nuclear proteins were also extracted and analyzed by Western blot. [1]
The MTS assay was used to assess cell proliferation. Cells were seeded in 96-well plates, treated with Xanthatin, and then MTS/PMS was added. Absorbance was measured at 490 nm to calculate the IC50. [5]
For cell morphology assessment, cells were treated with Xanthatin or DDP (positive control) and images were taken with an inverted microscope. [5]
Flow cytometry with PI staining was used for cell cycle analysis. Cells were treated with Xanthatin, harvested, fixed, and stained. The cell cycle distribution was determined by flow cytometry. [5]
For apoptosis analysis, cells were treated with Xanthatin and stained with FITC-labeled annexin V/PI, then analyzed by flow cytometry. Hoechst 33258 staining was also used to visualize nuclear changes. [5]
RT-PCR was performed to analyze gene expression. Total RNA was isolated, and cDNA was synthesized. PCR was performed with specific primers for p21, IL-1β, HO-1, GADD45α/β/γ, Cdc2, and cyclin B1. Products were separated by agarose gel electrophoresis. [4]
DNA fragmentation analysis was performed using a commercial kit. Cells were treated with (-)-Xanthatin, and DNA was extracted and analyzed by gel electrophoresis. Actinomycin D was used as a positive control for apoptosis. [4]
For LDH release assay, cells were treated with (-)-Xanthatin and the culture medium was analyzed for LDH using a non-radioactive cytotoxicity assay kit. [4]

For the in vivo test on murine ascite leukemia P-388, Xanthatin was injected intraperitoneally (i.p.) on days 1, 5, and 9. The study examined the T/C ratio at different doses. Signs of toxicity were observed by treating healthy mice with increasing doses. [3]

Xanthatin exhibited little or no toxicity to animals, with an LD50 value of 800 mg/kg. [3]
Signs of toxicity were observed at doses higher than 800 mg/kg in healthy mice. [3]

[1]. Molecules, 2012, 17(4):3736-50.
[2]. Lett Appl Microbiol, 1994, 18(4):206-8.
[3]. Planta Med, 1994, 60(5):473-4.
[4]. Chem Res Toxicol, 2011, 24(6):855-65.
[5]. Planta Med, 2012, 78(9):890-5.
[6]. Int J Clin Exp Pathol, 2015, 8(9):10355-64.

Xanthine is a sesquiterpene lactone. It has been reported to be found in cocklebur, Trichoderma dulcis, and other organisms for which relevant data exist.

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Xanthatin's antitumor activity is associated with the intrinsic apoptosis pathway and disruption of NF-κB signaling, suggesting it may have therapeutic potential against NSCLC. [1]
The study in MDA-MB-231 cells suggests that (-)-Xanthatin selectively induces GADD45γ, a tumor suppressor, leading to G2/M arrest and caspase-independent cell death. This effect is mediated through cellular stress pathways including oxidative stress and involves JNK and p38 MAPK signaling. [4]
Xanthatin may have therapeutic potential against human gastric carcinoma by inducing G2/M cell cycle arrest and apoptosis via the p53-mediated intrinsic pathway and NF-κB disruption. [5]

C15H18O3

246.3016

246.125

26791-73-1

5281511

Typically exists as solid at room temperature

1.1±0.1 g/cm3

444.3±45.0 °C at 760 mmHg

114.5-115°

199.1±28.8 °C

0.0±1.1 mmHg at 25°C

1.528

1.58

0

3

2

18

456

3

O1C(C(=C([H])[H])[C@@]2([H])C([H])([H])C([H])=C(/C(/[H])=C(\[H])/C(C([H])([H])[H])=O)[C@@]([H])(C([H])([H])[H])C([H])([H])[C@]12[H])=O

RBRPTFMVULVGIC-ZTIIIDENSA-N

InChI=1S/C15H18O3/c1-9-8-14-13(11(3)15(17)18-14)7-6-12(9)5-4-10(2)16/h4-6,9,13-14H,3,7-8H2,1-2H3/b5-4+/t9-,13+,14-/m0/s1

(3aR,7S,8aS)-7-methyl-3-methylidene-6-[(E)-3-oxobut-1-enyl]-4,7,8,8a-tetrahydro-3aH-cyclohepta[b]furan-2-one

Xanthatin; Xanthatin; 26791-73-1; (-)-Xanthatin; (3aR,7S,8aS)-7-Methyl-3-methylene-6-((E)-3-oxobut-1-en-1-yl)-3,3a,4,7,8,8a-hexahydro-2H-cyclohepta[b]furan-2-one; (3aR,7S,8aS)-7-methyl-3-methylidene-6-[(E)-3-oxobut-1-enyl]-4,7,8,8a-tetrahydro-3aH-cyclohepta[b]furan-2-one; CHEBI:10058; 298X1N12LS; CHEMBL404466;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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