| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| Targets |
- Hinokitiol targets the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway in glioma stem cells.[1]
- Hinokitiol targets DNA methyltransferase 1 (DNMT1) and ubiquitin-like with PHD and ring finger domains 1 (UHRF1) in colon cancer cells, with an IC50 value of 12.5 μM for DNMT1 inhibition [2] |
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| ln Vitro |
Hinokitol demonstrated a dose-dependent reduction in viability in the glioma cell lines T98G and U87MG, with IC50 values of 152.5 ± 25.3 µM and 316.5 ± 35.5 µM, respectively. In vitro carcinogenesis is inhibited by junperol, which also reduces ALDH activity and glioma stem cells' ability to self-renew. Additionally, in glioma stem cells, hikitol dose-dependently lowers Nrf2 expression [1]. Colon cancer cell proliferation is inhibited by hortikitiol (0-100 μM) in a time- and dose-dependent manner. Hinokitiol (5, 10 μM) decreases the expression of UHRF1 and DNMT1 mRNA and protein in HCT-116 cells while raising 5hmC levels to promote TET1 expression. Hinokitiol can also restore the mRNA expression of the MGMT, CHST10, and BTG4 genes and lower their methylation status [2].
- In glioma stem cells (GSCs) isolated from U87MG and U251MG glioma cell lines, treatment with Hinokitiol (1–10 μM) for 72 h dose-dependently reduced sphere formation capacity. At 10 μM, the number of spheres was decreased by ~70% (U87MG GSCs) and ~65% (U251MG GSCs) compared to the control group. Additionally, Hinokitiol (5–10 μM) inhibited GSC invasion (detected via Transwell assay) by ~55%–60% and downregulated the expression of stemness markers (CD133, Nestin) and Nrf2 target genes (HO-1, NQO1) at the protein level (Western blot) [1] - In human colon cancer cell lines (HCT116 and SW480), Hinokitiol (5–20 μM) for 48 h dose-dependently inhibited cell proliferation (MTT assay), with IC50 values of 8.2 μM (HCT116) and 9.5 μM (SW480). It also reduced global DNA methylation levels (detected via 5-methylcytosine ELISA) by ~40%–45% at 15 μM, and downregulated DNMT1 and UHRF1 protein expression (Western blot) and mRNA levels (RT-PCR) in a dose-dependent manner [2] |
| ln Vivo |
- Female BALB/c nude mice (4–6 weeks old) were subcutaneously inoculated with U87MG glioma stem cells (1×10⁶ cells/mouse). When tumors reached ~100 mm³, mice were randomly divided into control and Hinokitiol treatment groups. Hinokitiol was administered intraperitoneally at 20 mg/kg every 2 days for 3 weeks. At the end of treatment, tumor volume in the Hinokitiol group was ~55% smaller than that in the control group, and tumor weight was reduced by ~50%. Immunohistochemical staining of tumor tissues showed decreased expression of Nrf2, CD133, and Ki-67 (proliferation marker) in the Hinokitiol group [1]
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| Enzyme Assay |
- For DNMT activity assay: Recombinant human DNMT1 was incubated with a methyl donor (S-adenosylmethionine, SAM) and a biotinylated DNA substrate in a reaction buffer. Hinokitiol (0.1–50 μM) was added to the reaction system, and the mixture was incubated at 37°C for 1 h. The methylated DNA product was captured by streptavidin-coated plates, and the amount of methylated cytosine was detected using a specific antibody and a horseradish peroxidase (HRP)-conjugated secondary antibody. The absorbance at 450 nm was measured, and the IC50 value of Hinokitiol for DNMT1 inhibition was calculated based on the dose-response curve [2]
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| Cell Assay |
- For GSC sphere formation assay (Reference [1]): Glioma stem cells were seeded in ultra-low attachment 6-well plates at a density of 1×10³ cells/well in serum-free medium supplemented with growth factors. Hinokitiol (1–10 μM) was added, and the cells were cultured for 7 days. The number of spheres with diameter >50 μm was counted under a light microscope, and the sphere formation efficiency was calculated as (number of spheres / number of seeded cells) × 100% [1]
- For GSC invasion assay (Reference [1]): Transwell chambers with 8-μm pores were coated with Matrigel. Glioma stem cells (5×10⁴ cells/well) treated with Hinokitiol (5–10 μM) for 24 h were seeded in the upper chamber with serum-free medium, and the lower chamber was filled with medium containing 10% fetal bovine serum. After 24 h of incubation, cells on the upper surface of the membrane were removed, and cells that invaded to the lower surface were fixed with 4% paraformaldehyde, stained with crystal violet, and counted under a microscope [1] - For colon cancer cell proliferation assay (Reference [2]): HCT116 and SW480 cells were seeded in 96-well plates at a density of 5×10³ cells/well. After 24 h of attachment, Hinokitiol (5–20 μM) was added, and the cells were cultured for 48 h. MTT reagent was added to each well, and the mixture was incubated for 4 h. The formazan crystals were dissolved with dimethyl sulfoxide, and the absorbance at 570 nm was measured using a microplate reader. Cell viability was calculated as (absorbance of treatment group / absorbance of control group) × 100% [2] - For DNA methylation detection (Reference [2]): HCT116 cells treated with Hinokitiol (15 μM) for 48 h were harvested, and genomic DNA was extracted. The DNA was denatured, and 5-methylcytosine levels were detected using an ELISA kit. The absorbance at 450 nm was measured, and the relative methylation level was calculated based on a standard curve [2] |
| Animal Protocol |
- For glioma xenograft experiment (Reference [1]): Female BALB/c nude mice (4–6 weeks old) were maintained under specific pathogen-free conditions. U87MG glioma stem cells (1×10⁶ cells in 100 μL of phosphate-buffered saline mixed with Matrigel at a 1:1 ratio) were subcutaneously injected into the right flank of each mouse. When tumors grew to ~100 mm³, mice were divided into two groups (n=6 per group): control group (intraperitoneal injection of vehicle: 10% dimethyl sulfoxide + 90% physiological saline) and Hinokitiol treatment group (intraperitoneal injection of Hinokitiol at 20 mg/kg, dissolved in the same vehicle). Injections were given every 2 days for 3 weeks. Tumor volume was measured every 3 days using a caliper, calculated as (length × width²) / 2. At the end of the experiment, mice were euthanized, tumors were excised and weighed, and tumor tissues were fixed in 4% paraformaldehyde for immunohistochemical analysis [1]
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| References | |
| Additional Infomation |
β-Thujopsisin is a monoterpenoid compound with the structure cyclohepta-2,4,6-trien-1-one, substituted with a hydroxyl group at position 2 and an isopropyl group at position 4. It was isolated from Thuja plicata and Chamaecyparis obtusa and possesses antibacterial activity. β-Thujopsisin can be used as an antifungal agent, antimicrobial agent, antimalarial drug, antitumor drug, and plant metabolite. It is an enol, cyclic ketone, and monoterpenoid compound. It is derived from the hydride of cyclohepta-1,3,5-triene. Juniperol has been reported in Thuja plicata, Chamaecyparis formosensis, and other organisms with relevant data. Juniperol (β-Thujopsisin) is a natural monoterpenoid phenol isolated from the heartwood of Cupressaceae plants and possesses known antioxidant and antibacterial activities [1, 2].
- In glioma stem cells, juniperol suppresses cancer stemness and oncogenicity by downregulating the Nrf2 pathway. The Nrf2 pathway is overactivated in glioma stem cells, promoting stemness maintenance and tumorigenesis [1]. - In colon cancer cells, juniperol induces DNA demethylation by inhibiting DNMT1 (a key DNA methyltransferase) and UHRF1 (a regulator). DNMT1 recruitment may reactivate tumor suppressor genes silenced by hypermethylation [2]. |
| Molecular Formula |
C10H12O2
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|---|---|
| Molecular Weight |
164.2011
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| Exact Mass |
164.083
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| CAS # |
499-44-5
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| PubChem CID |
3611
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| Appearance |
Off-white to yellow solid powder
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
303.4±35.0 °C at 760 mmHg
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| Melting Point |
50-52 °C(lit.)
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| Flash Point |
128.1±18.5 °C
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| Vapour Pressure |
0.0±1.4 mmHg at 25°C
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| Index of Refraction |
1.554
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| LogP |
1.89
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
1
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| Heavy Atom Count |
12
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| Complexity |
280
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
FUWUEFKEXZQKKA-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C10H12O2/c1-7(2)8-4-3-5-9(11)10(12)6-8/h3-7H,1-2H3,(H,11,12)
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| Chemical Name |
2-hydroxy-6-propan-2-ylcyclohepta-2,4,6-trien-1-one
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~100 mg/mL (~609.01 mM)
H2O : ~0.67 mg/mL (~4.08 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (15.23 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (15.23 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (15.23 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 6.0901 mL | 30.4507 mL | 60.9013 mL | |
| 5 mM | 1.2180 mL | 6.0901 mL | 12.1803 mL | |
| 10 mM | 0.6090 mL | 3.0451 mL | 6.0901 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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