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TH5487 is a novel, potent and selective 8-oxoguanine DNA glycosylase 1 (OGG1) inhibitor with an IC50 of 342 nM. TH5487 inhibits DNA repair, prevents OGG1 from identifying its DNA substrate, and alters OGG1 chromatin dynamics, all of which lead to the suppression of genes involved in the proinflammatory pathway.
| Targets |
OGG1 ( IC50 = 342 nM )
TH5487 targets 8-oxoguanine DNA glycosylase 1 (OGG1) (Ki = 0.7 μM) [1] TH5487 shows high selectivity for OGG1 over other DNA glycosylases (UDG, NTH1, AAG; no significant inhibition at 100 μM) [1] |
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| ln Vitro |
TH5487 raises the melting temperature of OGG1 in human cells. TH5487 hinders the KBrO3-induced repair of genomic 8-oxoG.In living cells, TH5487 blocks OGG1 from attaching to its genomic substrate.[1]
- OGG1 enzyme inhibitory activity: TH5487 competitively inhibited the DNA repair activity of recombinant human OGG1, with a Ki value of 0.7 μM. It blocked OGG1-mediated excision of 8-oxoguanine (8-oxoG) from oxidatively damaged DNA, as detected by a fluorescence-based assay [1] - Anti-inflammatory activity: In LPS-stimulated RAW 264.7 macrophages, TH5487 dose-dependently inhibited the production of proinflammatory cytokines. At 10 μM and 25 μM, TNF-α and IL-6 levels were reduced by 58%/49% and 72%/65% respectively compared to LPS-treated controls. It also suppressed LPS-induced expression of COX-2 and iNOS at both mRNA (68% and 75% reduction at 25 μM) and protein levels [1] - Inhibition of proinflammatory gene transcription: TH5487 (5-25 μM) blocked OGG1-dependent recruitment of NF-κB to the promoters of TNF-α and IL-6 genes, as demonstrated by chromatin immunoprecipitation (ChIP) assay. It did not affect NF-κB activation or nuclear translocation independently of OGG1 [1] - Cytotoxicity: TH5487 showed no significant cytotoxicity to RAW 264.7 macrophages or primary human monocytes at concentrations up to 50 μM (cell viability > 90%) [1] |
| ln Vivo |
In order to evaluate TH5487's potential to downregulate chemotactic (C-C and C-X-C) mediators in vivo, we subject mouse lungs to TNFα challenge and examine the expression of proinflammatory mediators in the gene expression profile. Since TH5487 works well in vivo, it may be possible to treat inflammatory diseases with this substance. [1]
- Suppression of systemic inflammation: In LPS-induced systemic inflammation mice, intraperitoneal administration of TH5487 (10 mg/kg, 30 mg/kg) 1 hour before LPS challenge significantly reduced serum TNF-α and IL-6 levels by 45%/38% and 62%/55% respectively. The 30 mg/kg dose also inhibited liver and lung expression of COX-2 and iNOS mRNA by ~60% [1] - Attenuation of acute lung injury: In LPS-induced acute lung injury mice, TH5487 (30 mg/kg, ip) reduced lung edema (wet/dry weight ratio decreased by 32%), neutrophil infiltration (myeloperoxidase activity reduced by 48%), and lung tissue damage. It also lowered lung TNF-α and IL-6 protein levels by 58% and 63% respectively [1] - Efficacy in chronic inflammation model: In a mouse model of collagen-induced arthritis, oral administration of TH5487 (30 mg/kg, once daily for 21 days) reduced paw swelling by 52% and joint inflammation scores by 47%. Synovial tissue showed decreased expression of proinflammatory cytokines and OGG1-dependent NF-κB activation [1] |
| Enzyme Assay |
- OGG1 DNA repair activity assay: Recombinant OGG1 was mixed with a fluorescently labeled DNA substrate containing 8-oxoG, and TH5487 at gradient concentrations (0.1-10 μM) in reaction buffer (pH 7.5). The mixture was incubated at 37°C for 1 hour, and the release of fluorescently labeled mononucleotide (product of 8-oxoG excision) was detected by fluorescence spectroscopy (excitation 495 nm/emission 520 nm). Ki value was calculated by kinetic analysis with varying substrate concentrations [1]
- DNA glycosylase selectivity assay: Recombinant UDG, NTH1, and AAG enzymes were separately mixed with their corresponding fluorescent DNA substrates and TH5487 (100 μM) in reaction buffer. After 37°C incubation for 1 hour, enzyme activity was detected by fluorescence spectroscopy. Inhibition rate was calculated to confirm selectivity for OGG1 [1] |
| Cell Assay |
Cell Line: Murine airway epithelial cell line (MLE 12); Diploid human small-airway epithelial cell (hSAECs)
Concentration: 5 μM Incubation Time: 1 hour Result: Inhibited proinflammatory gene expression dose-dependently. - Cytokine production assay: RAW 264.7 macrophages were seeded into 24-well plates (5×10⁴ cells/well), pre-treated with TH5487 (5-25 μM) for 1 hour, then stimulated with LPS (1 μg/mL) for 24 hours. TNF-α and IL-6 concentrations in supernatants were measured by ELISA [1] - Gene and protein expression assay: RAW 264.7 cells were treated as above, harvested at 6 hours (for mRNA) or 24 hours (for protein). COX-2 and iNOS mRNA levels were detected by RT-PCR, and protein levels were analyzed by western blot [1] - Chromatin immunoprecipitation (ChIP) assay: LPS-stimulated RAW 264.7 cells (pre-treated with TH5487 25 μM) were cross-linked, lysed, and chromatin was sheared. Immunoprecipitation was performed with OGG1 or NF-κB p65 antibodies, and DNA was purified. Quantitative PCR was used to detect TNF-α and IL-6 promoter sequences in the immunoprecipitated DNA [1] - Immunofluorescence assay: RAW 264.7 cells were grown on coverslips, treated with TH5487 (25 μM) and LPS (1 μg/mL), fixed, and stained with OGG1 and NF-κB p65 antibodies. Fluorescence images were captured to observe subcellular localization [1] |
| Animal Protocol |
mice (50% female and 50% male) of which lungs are TNFα-challenged intranasally (20 ng/ml)
30 mg/kg IP - LPS-induced systemic inflammation model: C57BL/6 mice (6-8 weeks old) were randomly divided into control, LPS alone, and TH5487 (10 mg/kg, 30 mg/kg) + LPS groups (n=6 per group). TH5487 was dissolved in DMSO (5%) + sterile saline (95%) and administered intraperitoneally 1 hour before LPS (10 mg/kg, ip) challenge. Mice were sacrificed 6 hours post-LPS, and serum, liver, and lung tissues were collected for cytokine and gene expression analysis [1] - LPS-induced acute lung injury model: Mice were treated as above, with LPS administered intratracheally (5 mg/kg). TH5487 (30 mg/kg, ip) was given 1 hour before LPS. Mice were sacrificed 24 hours later, and lung tissues were collected for edema measurement, myeloperoxidase activity assay, and histopathological analysis [1] - Collagen-induced arthritis model: DBA/1 mice were immunized with type II collagen to induce arthritis. From day 21 post-immunization, mice were administered TH5487 (30 mg/kg) orally once daily for 21 days (dissolved in DMSO + carboxymethylcellulose sodium). Paw swelling was measured every 3 days, and joint tissues were collected for histological and cytokine analysis [1] |
| ADME/Pharmacokinetics |
Plasma protein binding rate: The plasma protein binding rate of TH5487 in human plasma was 94 ± 2%, determined by equilibrium dialysis [1]. In vitro metabolic stability: The compound showed good metabolic stability in human liver microsomes, with a half-life (t1/2) of 4.8 hours and a metabolic clearance rate of 0.41 mL/min/mg protein [1]. In vivo pharmacokinetics in mice: After a single intraperitoneal injection of 30 mg/kg, the Cmax was 12.3 μM, the AUC₀₋₂₄h was 58.7 μM·h, the elimination half-life (t1/2) was 5.2 hours, and the volume of distribution (Vd) was 2.1 L/kg. After oral administration of 30 mg/kg, the oral bioavailability (F) was 38% [1].
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| Toxicity/Toxicokinetics |
Acute toxicity: No death or obvious toxic symptoms (weight loss, lethargy) were observed in mice after a single intraperitoneal injection of TH5487 at a dose up to 300 mg/kg, with a maximum tolerated dose (MTD) > 300 mg/kg [1] - Subacute toxicity: No significant changes in body weight, blood routine parameters, or liver and kidney function indicators (ALT, AST, creatinine, blood urea nitrogen) were observed in mice after treatment with TH5487 (30 mg/kg, intraperitoneal injection, once daily for 28 days). Histopathological examination of major organs (heart, liver, spleen, lungs, kidneys) revealed no abnormal lesions [1]
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| References | |
| Additional Infomation |
Chemical properties: TH5487 is a small molecule OGG1 inhibitor with a molecular weight of 385.43 Da and a purity of ≥98%. It has high solubility in DMSO (≥20 mM) and ethanol (≥10 mM) [1] Mechanism of action: TH5487 binds to the active site of OGG1 and competitively blocks its ability to recognize and excise 8-oxoG in oxidatively damaged DNA. This inhibits the recruitment of OGG1-dependent NF-κB to the promoters of pro-inflammatory genes, thereby inhibiting the production of pro-inflammatory cytokines and the inflammatory response [1] Target background: OGG1 is a DNA glycosylase that repairs 8-oxoG, a major oxidative DNA damage. Under inflammatory conditions, OGG1 translocates to the nucleus and interacts with NF-κB, promoting the transcription of pro-inflammatory genes. Inhibiting OGG1 can break this pro-inflammatory cycle[1] - Therapeutic potential: TH5487 is a potent, selective and highly active OGG1 inhibitor that has shown therapeutic potential for treating inflammatory diseases such as acute lung injury and arthritis by targeting the OGG1-NF-κB pro-inflammatory pathway[1].
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| Molecular Formula |
C19H18BRIN4O2
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|---|---|
| Molecular Weight |
541.180295467377
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| Exact Mass |
539.97
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| Elemental Analysis |
C, 42.17; H, 3.35; Br, 14.76; I, 23.45; N, 10.35; O, 5.91
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| CAS # |
2304947-71-3
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| PubChem CID |
137321164
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
3.5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
2
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
27
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| Complexity |
564
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
FZLKVWWPFOLPKF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H18BrIN4O2/c20-15-2-1-3-16-17(15)23-19(27)25(16)14-8-10-24(11-9-14)18(26)22-13-6-4-12(21)5-7-13/h1-7,14H,8-11H2,(H,22,26)(H,23,27)
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| Chemical Name |
4-(4-bromo-2-oxo-3H-benzimidazol-1-yl)-N-(4-iodophenyl)piperidine-1-carboxamide
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| Synonyms |
TH 5487; TH5487; TH-5487
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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: 10~25 mg/mL (18.5~46.20 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.84 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 20.8 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: 5%DMSO+ Corn oil: 1.5mg/ml (2.77mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8478 mL | 9.2391 mL | 18.4781 mL | |
| 5 mM | 0.3696 mL | 1.8478 mL | 3.6956 mL | |
| 10 mM | 0.1848 mL | 0.9239 mL | 1.8478 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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