Tedizolid (Synonyms: DA 7157, TR-700)

Cell lines

NCM460 cells

Preparation Method

NCM460 cells were cultured in a humidified incubator with 5% CO₂ in complete medium containing Roswell Park Memorial Institute 1640 medium (RPMI-1640), 10% fetal bovine serum (FBS), 100units/ml penicillin, 0.1mg/ml streptomycin, and 2mM L-glutamine. Dextran sulfate sodium (DSS) was used in induction of cellular senescence. Briefly, NCM460 were seeded in 24-well plates, when the confluence reached 50%, DSS was added in the complete medium for 4 days. For optimization test, 1-10μg/ml DSS was used, and 3μg/ml was selected as the optimum concentration for cellular senescence model construction. Cell viability and drug toxicity were determined by cell counting Kit. 1000 NCM460 cells were seeded in 96 well plates with 100μl complete medium. After 24h, different concentrations of DSS or DSS and Tedizolid (0-10μM) were added and incubated for 72h. Subsequently, the medium was changed to 100μl working solution containing 10μl CCK-8 reagent and 90μl complete medium and incubated for 0.5-1h, cell viability was measured with a microplate reader at 450nm. Senescence-associated β-galactosidase (SA-β-gal) activity was detected using the SA-β-gal staining kit following the manufacturer’s instructions. Cells and colonic tissues were collected for Western blot and Quantitative real-time PCR analyses.

Reaction Conditions

0-10μM; 72h

Applications

Treatment of human colonic epithelial NCM460 cells with Tedizolid significantly reversed DSS-induced senescence, reduced SA-β-gal-positive cells, downregulated P16 and P21 expression, and restored p-AMPK levels.

Animal models

female ICR mice

Preparation Method

Specific-pathogen-free, female ICR mice weighing approximately 20-25g were provided food and water ad libitum. Mice was rendered transiently neutropenic by injecting cyclophosphamide intraperitoneal at a dose of 150mg/kg of body weight at four days before inoculation and 100mg/kg of body weight at one day before inoculation. Isolates to be inoculated into the thighs were previously frozen at -80℃ in skim milk. Two transfers of the organism was performed onto Trypticase Soy Agar plates with 5% sheep blood and Burusera HK nutrient agar, and placed into an incubator at 37℃ for approximately 24h. After an 18-24h incubation of the 2nd transfer, a bacterial suspension of approximately 10⁷CFU/ml was made for inoculation. Final inoculum concentrations were confirmed by serial dilution and plating techniques. Thigh infection with each of the test isolates will be produced by intramuscular injection of 0.1ml of the inoculum into each thigh of the mice 2h prior to the initiation of antimicrobial therapy. To assess the in vivo activity of human simulated Tedizolid (200mg daily) against MRSA and P. anaerobius exhibiting various Phenotypic, Tedizolid 200mg daily was given intraperitoneal at 0h. Control animals received sterile normal saline in the same volume, route, and schedule as the active drug regimen. All animals were euthanized by CO₂ exposure followed by cervical dislocation by 24h as appropriate. After sacrifice, the thighs were removed and individually homogenized in normal saline. Serial dilutions were plated on an appropriate agar media for CFU/ml determination.

Dosage form

200mg daily for 24h; i.p.

Applications

Tedizolid significantly reduced MRSA thigh burden in neutropenic mouse mice with mixed P.anaerobius infection.

References:
[1] Zhou M, Liu ZL, Liu JY, Wang XB. Tedizolid phosphate alleviates DSS-induced ulcerative colitis by inhibiting senescence of cell and colon tissue through activating AMPK signaling pathway. Int Immunopharmacol. 2024;135:112286.
[2] Yamagishi Y, Mikamo H, Kato H, et al. Efficacy of tedizolid against methicillin-resistant Staphylococcus aureus and Peptostreptococcus anaerobius in thigh mixed-infection mouse model. J Infect Chemother. 2017;23(6):368-373.