Thymoquinone targets multiple pathways including VEGFR2-PI3K-Akt, STAT3, JNK, p38, 5-LO, COX, and PGD2. By downregulating the VEGFR2-PI3K-Akt pathway, the compound inhibits angiogenesis and tumor growth. By suppressing STAT3 activation, it inhibits proliferation and induces apoptosis in cancer cells. Through the activation of JNK and p38, it regulates apoptosis and other cellular processes. Thymoquinone also inhibits 5-LO and COX enzymes, which are involved in the production of inflammatory mediators. The compound's multi-targeted activity makes it a valuable tool for studying various diseases, including cancer, inflammation, and neurodegenerative disorders.

In vitro, Thymoquinone inhibits the growth of leukemia cells with an IC50 of 1.5 µg/ml. It induces apoptosis by down-regulating Bcl-2 while up-regulating Bax expression. The compound inhibits proliferation, induces apoptosis, and chemosensitizes human multiple myeloma cells through suppression of STAT3 activation. It induces mitochondria-mediated apoptosis in acute lymphoblastic leukemia and regulates apoptosis through JNK and p38 activation. Thymoquinone (25–100 µM) is used to treat human breast cancer cell lines. The compound's in vitro activity is well-characterized and forms the basis for its use in cancer and inflammation research.

In vivo, Thymoquinone demonstrates anticancer, anti-inflammatory, and hepatoprotective activities. It synergizes with 5-FU to increase anticancer efficiency against triple-negative breast cancer (TNBC) cells. The combination of DFMO and TQ shows promise for treating acute T lymphoblastic leukemia. Thymoquinone has been studied in animal models of Alzheimer's disease, cardiovascular disease, and infections. The compound's in vivo efficacy is attributed to its ability to modulate multiple signaling pathways and exert antioxidant, anti-inflammatory, and anticancer effects. Clinical trials have been conducted to evaluate the therapeutic potential of Thymoquinone in various human conditions, including type 2 diabetes, cancer, COVID-19, epilepsy, and chronic periodontitis.

Cell-free assays with Thymoquinone are used to evaluate its effects on various targets. Kinase activity assays measure inhibition of VEGFR2, PI3K, Akt, and STAT3. COX and 5-LO enzyme activities are assessed using standard enzymatic assays. Antioxidant activity is measured by DPPH or ABTS radical scavenging assays. These cell-free assays are essential for characterizing the potency and selectivity of Thymoquinone for its various targets.

In cellular assays, cancer cell lines are treated with Thymoquinone at varying concentrations. Cell viability is assessed using MTT assays. Apoptosis is evaluated by flow cytometry (Annexin V/PI staining) and caspase activity assays. Inflammatory cytokine production is measured by ELISA. These cellular assays are crucial for understanding the functional consequences of Thymoquinone treatment and for validating its activity as an anticancer and anti-inflammatory agent.

In vivo efficacy of Thymoquinone is evaluated in mouse xenograft models of various cancers. The compound is administered orally or via intraperitoneal injection. Tumor growth, angiogenesis, inflammatory markers, and survival are monitored. The compound has been studied in models of Alzheimer's disease, cardiovascular disease, and infections. These in vivo studies are essential for confirming the compound's efficacy in a physiologically relevant context and for guiding the development of Thymoquinone as a therapeutic agent. Clinical trials have been conducted to evaluate its therapeutic potential in various human conditions.

Pharmacokinetic properties of Thymoquinone including bioavailability, half-life, and tissue distribution are determined in preclinical studies. The compound has low oral bioavailability due to poor solubility and extensive metabolism. Various formulations have been developed to improve PK. The pharmacokinetic data obtained from these studies are essential for designing appropriate dosing regimens and for interpreting the results of efficacy and toxicity studies.

Toxicological studies indicate that Thymoquinone has a favorable safety profile at therapeutic doses. High doses may cause hepatotoxicity. Standard toxicology studies in animals evaluate organ toxicity, hematological parameters, and therapeutic window. The compound is generally well-tolerated in clinical trials. Standard safety precautions should be followed when handling Thymoquinone, including the use of appropriate personal protective equipment and adherence to institutional safety guidelines.

[1]. Protective role of thymoquinone against paraquat-induced hepatotoxicity in mice. Pestic Biochem Physiol. 2018 Jun;148:16-21.

[2]. Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: a possible mechanism of protection. Pharmacol Res. 2000 Mar;41(3):283-9.

[3]. Thymoquinone and cisplatin as a therapeutic combination in lung cancer: In vitro and in vivo. J Exp Clin Cancer Res. 2010 Jul 1;29(1):87.

[4]. Thymoquinone administration ameliorates Alzheimer's disease-like phenotype by promoting cell survival in the hippocampus of amyloid beta1-42 infused rat model. Phytomedicine. 2020 Dec;79:153324.

[5]. Thymoquinone inhibits inflammation, neoangiogenesis and vascular remodeling in asthma mice. Int Immunopharmacol. 2016 Sep;38:70-80.

Thymicin belongs to the 1,4-benzoquinone class of compounds, meaning that the hydrogen atoms at positions 2 and 5 of the 1,4-benzoquinone molecule are replaced by methyl and isopropyl groups, respectively. It is a natural compound isolated from Nigella sativa and has shown good chemotherapeutic activity. Thymicin possesses various effects including anti-inflammatory, antioxidant, adjuvant, antitumor, cardioprotective, antidepressant, and phytometabolic activity. Thymicin is a natural compound with broad protective effects, including antioxidant, anti-inflammatory, immunomodulatory, anticancer, and antibacterial activities. It has been reported to exist in oregano (Origanum dictamnus), Ranunculaceae plants, and other organisms with relevant data. Mechanism of Action: Thymicin is a natural compound with broad protective effects, including antioxidant, anti-inflammatory, immunomodulatory, anticancer, and antibacterial activities. It can induce apoptosis, regulate pro-apoptotic and anti-apoptotic genes, and inhibit cancer metastasis by activating JNK and p38. Its anti-inflammatory effect is attributed to its inhibition of inflammatory cytokines and processes, including pathways associated with 5-LO, COX, and PGD2.
Thymoquinone (TQ, CAS 490-91-5) is a natural product and research compound. It is derived from Nigella sativa (black seed) and possesses diverse pharmacological activities including antioxidant, anti-inflammatory, anticancer, antiviral, anticonvulsant, antifungal, antiangiogenic, and hepatoprotective effects. Thymoquinone is used in research related to Alzheimer's disease, cancer, cardiovascular diseases, infectious diseases, and inflammation. It has been evaluated in clinical trials for various conditions. The compound is not approved for human therapeutic use and is intended for research purposes only. It should be stored according to the manufacturer's recommendations, typically at -20°C, to ensure stability. When handling Thymoquinone, researchers should follow standard safety protocols for handling chemical reagents, including the use of appropriate personal protective equipment and working in a well-ventilated area.

C10H12O2

164.2011

164.083

490-91-5

10281

Light yellow to yellow solid powder

1.1±0.1 g/cm3

232.0±15.0 °C at 760 mmHg

167 ºC

103.9±0.0 °C

0.1±0.5 mmHg at 25°C

1.505

2.33

0

2

1

12

293

0

KEQHJBNSCLWCAE-UHFFFAOYSA-N

InChI=1S/C10H12O2/c1-6(2)8-5-9(11)7(3)4-10(8)12/h4-6H,1-3H3

2-methyl-5-propan-2-ylcyclohexa-2,5-diene-1,4-dione

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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 (~609.01 mM)
H2O : ~1 mg/mL (~6.09 mM)

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.

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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.

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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.
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.)