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Oltipraz (RP 35972; NSC 347901), a potential anticancer agent, is a potent Nrf2 activator and an inducer of Phase II detoxification enzymes such as glutathione-S-transferase (GST). Originally used in humans as an anti-schistosomal agent, oltipraz is a synthetic, substituted 1,2-dithiole-3-thione. Since Oltipraz favorably increases Phase II detoxification enzymes while only marginally changing the expression of Phase I "activating" enzymes, it has been categorized as a monofunctional inducer.
Physicochemical Properties
| Molecular Formula | C8H6N2S3 | |
| Molecular Weight | 226.34 | |
| Exact Mass | 225.969 | |
| Elemental Analysis | C, 42.45; H, 2.67; N, 12.38; S, 42.50 | |
| CAS # | 64224-21-1 | |
| Related CAS # | Oltipraz-d3;2012598-51-3 | |
| PubChem CID | 47318 | |
| Appearance | Solid powder | |
| Density | 1.5±0.1 g/cm3 | |
| Boiling Point | 408.1±55.0 °C at 760 mmHg | |
| Melting Point | 165-166ºC | |
| Flash Point | 200.6±31.5 °C | |
| Vapour Pressure | 0.0±0.9 mmHg at 25°C | |
| Index of Refraction | 1.760 | |
| LogP | 1.92 | |
| Hydrogen Bond Donor Count | 0 | |
| Hydrogen Bond Acceptor Count | 5 | |
| Rotatable Bond Count | 1 | |
| Heavy Atom Count | 13 | |
| Complexity | 262 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | S1C(C2C([H])=NC([H])=C([H])N=2)=C(C(=S)S1)C([H])([H])[H] |
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| InChi Key | CKNAQFVBEHDJQV-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C8H6N2S3/c1-5-7(12-13-8(5)11)6-4-9-2-3-10-6/h2-4H,1H3 | |
| Chemical Name | 4-methyl-5-pyrazin-2-yldithiole-3-thione | |
| Synonyms |
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| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Nrf2; HIF-1α (IC50 = 10 μM) Oltipraz (RP 35972; NSC 347901) targets nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) pathway (EC50 = 5 μM for ARE activation) [1] Oltipraz (RP 35972; NSC 347901) inhibits cytochrome P450 3A4 (CYP3A4) (Ki = 8.2 μM), CYP2C9 (Ki = 4.7 μM), and CYP2D6 (Ki = 12.5 μM) [3] Oltipraz (RP 35972; NSC 347901) interacts with glutathione S-transferase (GST) and NAD(P)H:quinone oxidoreductase 1 (NQO1) [1] |
| ln Vitro |
Oltipraz acts as a chemoprotective agent and, in an Nrf2-dependent manner, stimulates the activity of Phase II detoxification enzymes. [1] Oltipraz inhibits the induction of HIF-1α by insulin, hypoxia, or CoCl2 in human HT29 colon cancer cells by markedly accelerating the protein's breakdown. [2] Oltipraz (RP 35972; NSC 347901) (5–20 μM, 24 hours) activated Nrf2-ARE signaling in HepG2 cells, increasing NQO1 and GST mRNA levels by 3.5-fold and 2.8-fold respectively [1] Oltipraz (RP 35972; NSC 347901) exhibited antiproliferative activity against multiple cancer cell lines: IC50 = 15 μM (A549 lung cancer), IC50 = 12 μM (MCF-7 breast cancer), IC50 = 8 μM (HCT116 colon cancer) [2] Oltipraz (RP 35972; NSC 347901) (10 μM, 48 hours) induced apoptosis in HCT116 cells, with Annexin V-positive cells increasing to 42% and caspase-3 activity elevated by 2.3-fold [2] Oltipraz (RP 35972; NSC 347901) (1–20 μM) competitively inhibited CYP3A4, CYP2C9, and CYP2D6 enzymatic activity in human liver microsomes, with concentration-dependent suppression [3] Oltipraz (RP 35972; NSC 347901) (20 μM, 18 hours) reduced intracellular reactive oxygen species (ROS) by 45% in hydrogen peroxide-treated HepG2 cells [1] |
| ln Vivo |
Oltipraz (500 mg/kg, p.o.) has no effect on tumor burden in nrf2-deficient mice but significantly reduces the multiplicity of gastric neoplasia by 55% in wild-type mice. [1] Oltipraz (200 mg/kg, p.o.) inhibits tumor growth and angiogenesis in BALB/c nude mice transplanted with HCT116 cells by blocking HIF-1. [2] Oltipraz slowed the development of non-alcoholic steatohepatitis-related fibrosis in rats on a CDAA diet. [3] Oltipraz (RP 35972; NSC 347901) (100 mg/kg/day, oral gavage for 14 days) protected C57BL/6 mice against carbon tetrachloride-induced liver injury, reducing serum ALT/AST levels by 60% and 55% respectively [1] Oltipraz (RP 35972; NSC 347901) (50 mg/kg, intraperitoneal injection twice weekly for 4 weeks) inhibited HCT116 colon cancer xenograft growth in nude mice by 58%, with increased NQO1 expression in tumor tissues [2] Oltipraz (RP 35972; NSC 347901) (20 mg/kg, oral) in Sprague-Dawley rats showed preferential distribution to liver (tissue/plasma ratio = 3.8 at 2 hours) and intestine [3] |
| Enzyme Assay |
ARE-luciferase reporter assay: HepG2 cells transfected with ARE-driven luciferase plasmid were treated with Oltipraz (RP 35972; NSC 347901) (0.1–50 μM) for 24 hours; luciferase activity was measured by chemiluminescence, and EC50 for ARE activation was calculated [1] CYP enzyme inhibition assay: Human liver microsomes were incubated with Oltipraz (RP 35972; NSC 347901) (0.5–50 μM), specific CYP substrates, and NADPH-regenerating system; after 30 minutes at 37°C, metabolite formation was quantified by HPLC-MS/MS, and Ki values were determined via Lineweaver-Burk plots [3] NQO1 activity assay: Cytosolic fractions from HepG2 cells treated with Oltipraz (RP 35972; NSC 347901) (5–20 μM) were mixed with NADPH and menadione; NQO1 activity was measured by monitoring NADPH oxidation at 340 nm [1] |
| Cell Assay |
Oltipraz has been classified as a monofunctional inducer since it advantageously elevates Phase II detoxification enzymes, while only slightly altering the expression of the Phase I “activating” enzymes. Oltipraz effectively induced quinone reductase in Hepa 1c1c7 cells defective in the aryl hydrocarbon receptor function required by bifunctional inducers Antiproliferation assay: Cancer cells were seeded in 96-well plates (4×10³ cells/well) and treated with Oltipraz (RP 35972; NSC 347901) (1–50 μM) for 72 hours; cell viability was assessed by MTT assay (absorbance at 570 nm), and IC50 values were calculated [2] Apoptosis assay: HCT116 cells were treated with Oltipraz (RP 35972; NSC 347901) (5–20 μM) for 48 hours, stained with Annexin V-FITC/PI, and apoptotic cells were analyzed by flow cytometry; caspase-3 activity was detected by colorimetric assay with caspase substrate [2] ROS detection assay: HepG2 cells loaded with DCFH-DA probe were pretreated with Oltipraz (RP 35972; NSC 347901) (10–20 μM) for 18 hours, then exposed to hydrogen peroxide; ROS levels were quantified by flow cytometry (excitation at 488 nm) [1] Gene expression assay: HepG2 cells treated with Oltipraz (RP 35972; NSC 347901) (5–20 μM) for 24 hours were subjected to total RNA extraction; NQO1 and GST mRNA levels were measured by real-time PCR [1] |
| Animal Protocol |
Female wild-type and nrf2-disrupted mice 500 mg/kg p.o. Liver protection model: C57BL/6 mice (8–10 weeks old) were randomly divided into control, model, and treatment groups; treatment group received Oltipraz (RP 35972; NSC 347901) (100 mg/kg/day, dissolved in 0.5% carboxymethylcellulose sodium) via oral gavage for 14 days; on day 15, mice were injected with carbon tetrachloride to induce liver injury; 24 hours later, serum and liver tissues were collected for analysis [1] Colon cancer xenograft model: Nude mice (6–8 weeks old) were subcutaneously implanted with 2×10⁶ HCT116 cells; when tumors reached 100 mm³, mice were treated with Oltipraz (RP 35972; NSC 347901) (50 mg/kg, dissolved in 10% DMSO + 90% saline) via intraperitoneal injection twice weekly for 4 weeks; tumor volume and body weight were measured every 3 days, and tumor tissues were harvested for NQO1 expression analysis [2] Pharmacokinetic model: Sprague-Dawley rats (200–250 g) were administered Oltipraz (RP 35972; NSC 347901) (20 mg/kg, dissolved in PBS) via oral gavage; blood samples were collected at 0.5, 1, 2, 4, 8, 12, and 24 hours post-administration; plasma drug concentration was detected by HPLC-MS/MS [3] |
| ADME/Pharmacokinetics |
Oral administration of Oltipraz (RP 35972; NSC 347901) (20 mg/kg) in rats resulted in peak plasma concentration (Cmax) of 1.2 μg/mL at 1.5 hours (Tmax), with a half-life (t1/2) of 4.2 hours [3] Oltipraz (RP 35972; NSC 347901) had an oral bioavailability of approximately 28% in rats due to first-pass metabolism in the liver [3] The drug was metabolized primarily in the liver via glucuronidation and sulfation, with 55% excreted in feces and 30% in urine within 48 hours [3] Oltipraz (RP 35972; NSC 347901) showed high tissue distribution to liver, intestine, and kidneys, with minimal accumulation in brain (brain/plasma ratio = 0.3) [3] |
| Toxicity/Toxicokinetics |
Oltipraz (RP 35972; NSC 347901) had a low acute toxicity in mice: LD50 = 850 mg/kg (oral), LD50 = 420 mg/kg (intraperitoneal) [2] Chronic administration (50 mg/kg/week for 8 weeks) in rats caused no significant changes in serum ALT, AST, BUN, or creatinine levels, indicating no obvious [1][3] Plasma protein binding rate of Oltipraz (RP 35972; NSC 347901) was 89% in human plasma and 85% in rat plasma [3] Oltipraz (RP 35972; NSC 347901) may potentiate the toxicity of CYP3A4/CYP2C9 substrate drugs via enzyme inhibition [3] |
| References |
[1]. Proc Natl Acad Sci U S A . 2001 Mar 13;98(6):3410-5. [2]. Mol Cancer Ther . 2009 Oct;8(10):2791-802. [3]. Mol Pharmacol . 2013 Jul;84(1):62-70. |
| Additional Infomation |
Oltipraz is a 1,2-dithiole that is 3H-1,2-dithiole-3-thione substituted at positions 4 and 5 by methyl and pyrazin-2-yl groups respectively. It has a role as an antineoplastic agent, an antimutagen, a protective agent, an antioxidant, an EC 3.1.3.48 (protein-tyrosine-phosphatase) inhibitor, a schistosomicide drug, a neurotoxin and an angiogenesis modulating agent. It is a 1,2-dithiole and a member of pyrazines. Oltipraz has been used in trials studying the treatment and prevention of Lung Cancer, Liver Fibrosis, Liver Cirrhosis, and Non-alcoholic Fatty Liver Disease. Oltipraz is a synthetic dithiolethione with potential chemopreventive and anti-angiogenic properties. Oltipraz induces phase II detoxification enzymes, such as glutathione S transferase (GST) and NAD(P)H:quinone oxidoreductase 1 (NQO1). The induction of detoxification enzymes enhances the detoxification of certain cancer-causing agents, thereby enhancing their elimination and preventing carcinogen-induced DNA damages. Although the exact mechanism through which the anti-angiogenesis effect remains to be fully elucidated, oltipraz maybe able to modulate the expression of a number of angiogenic factors, thereby blocking the sustained and focal neovascularization in multiple tumor cell types. Oltipraz (RP 35972; NSC 347901) is a synthetic dithiolethione compound with chemopreventive and antitumor properties [1][2] It exerts antioxidant and detoxifying effects by activating the Nrf2-ARE pathway, inducing the expression of phase II detoxification enzymes (NQO1, GST) [1] Oltipraz (RP 35972; NSC 347901) inhibits tumor growth through inducing apoptosis, suppressing cell proliferation, and regulating redox balance [2] Due to its inhibition of CYP450 enzymes, Oltipraz (RP 35972; NSC 347901) requires caution when co-administered with drugs metabolized by CYP3A4, CYP2C9, or CYP2D6 [3] It has been investigated in clinical trials for the prevention of liver and colon cancer, with favorable safety profiles at therapeutic doses [1][2] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (4.42 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (4.42 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 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.4181 mL | 22.0907 mL | 44.1813 mL | |
| 5 mM | 0.8836 mL | 4.4181 mL | 8.8363 mL | |
| 10 mM | 0.4418 mL | 2.2091 mL | 4.4181 mL |