N-PHENYL-4-[[(PHENYLAMINO)THIOXOMETHYL]AMINO]-BENZENESULFONAMIDE

N-PHENYL-4-[[(PHENYLAMINO)THIOXOMETHYL]AMINO]-BENZENESULFONAMIDE Property

Boiling point:

549.4±60.0 °C(Predicted)

Density 

1.437±0.06 g/cm3(Predicted)

storage temp. 

Sealed in dry,2-8°C

solubility 

insoluble in EtOH; insoluble in H2O; insoluble in DMSO

form 

solid

pka

8.77±0.10(Predicted)

color 

White to off-white

Safety

Hazard Codes 

Xn

Risk Statements 

22

Hazard and Precautionary Statements (GHS)

N-PHENYL-4-[[(PHENYLAMINO)THIOXOMETHYL]AMINO]-BENZENESULFONAMIDE Chemical Properties,Usage,Production

Uses

LED209 is a potent and orally active small molecule inhibitor of the bacterial receptor QseC and a potent prodrug with high selectivity for QseC. LED209 inhibits the binding of signaling molecules to QseC. LED209 has antibacterial activity[1].

Biological Activity

many bacterial pathogens rely on a conserved membrane histidine sensor kinase, qsec, to respond to host adrenergic signaling molecules and bacterial signals in order to promote the expression of virulence factors. led209 is a potent prodrug that is highly selective for qsec.

Synthesis

General procedure for the synthesis of N-phenyl-4-(3-phenylthioureido)benzenesulfonamide from 4-amino-N-phenylbenzenesulfonamide and phenyl thioisocyanate: triethylamine (0.07 mL, 0.5 mmol) and phenyl thioisocyanate (2.1 mL, 11 mmol) were sequentially added to 4-amino-N-phenylbenzenesulfonamide (2.48 g, 10 mmol) in a anhydrous THF (10 mL) solution. The reaction mixture was heated under reflux conditions for 48 hours. After completion of the reaction, all volatiles were removed under vacuum and the crude product was purified by rapid chromatography on silica gel (eluent: hexane/acetone, 3:1) to afford N-phenyl-4-[[(phenylamino)thiomethyl]amino]benzenesulfonamide (Compound 5, also known as LED209) (1.34 g, 35% yield) as a white solid with a melting point of 160-162 °C. The 1H NMR (CD3COCD3, 300 MHz) δ of compound 5 9.35 (br s, 1H), 9.30 (br s, 1H), 8.98 (br s, 1H), 7.80-7.72 (m, 4H), 7.51-7.47 (m, 2H), 7.38-7.33 (m, 2H), 7.28-7.16 (m, 5H) , 7.08-7.04 (m, 1H); 13C NMR (CD3COCD3, 75 MHz) δ 180.2, 143.9, 138.9, 138.2, 135.2, 129.3 (2C), 129.1 (2C), 127.9 (2C), 125.8, 124.6 (3C), 122.8 (2C), and 120.8 (2C). Compound 27 was prepared using a similar preparation method as compound 3 in 90% yield; its 1H NMR (CD3COCD3, 300 MHz) δ 9.63 (br s, 1H), 7.80-7.77 (m, 2H), 7.46-7.42 (m, 2H), 7.35-7.27 (m, 3H), 7.15-7.12 (m, 2H), 7.15-7.12 (m, 2H) 3.17 (s, 3H), 2.14 (s, 3H). Compound 8 was prepared using a similar preparation method as compound 4 in 65% yield; its 1H NMR (CD3COCD3, 300 MHz) δ 7.32-7.13 (m, 7H), 6.68-6.65 (m, 2H), 5.53 (br s, 2H), 3.10 (s, 3H).

in vitro

led209-mediated inhibition of virulence gene expression inhibited the secretion of espa and espb, two proteins encoded within the locus of enterocyte effacement that are required for escherichia coli (ehec) to translocate bacterial proteins into host cells and cause attaching-effacing (ae) lesions. at 5 pm, led209 abolished ehec ae lesion formation on cultured epithelial cells. unlike conventional antibiotics, led209 does not kill or hinder ehec growth or trigger the ehec sos response [1].

in vivo

led209 [20 mg per kilogram of body weight (mg/kg)] was given orally to mice 3 hours before and 3 hours after intraperitoneal injection of a lethal dose of s. typhimurium. twenty-four hours after infection, only 30% of untreated mice remained alive, whereas 80% of the led209-treated mice were still alive. all the s. typhimurium– infected mice died within 72 hours of infection, and 20% of led209-treated mice survived up to 12 days [1].

IC 50

<10 μm

References

[1] rasko da, moreira cg, li de r, reading nc, ritchie jm, waldor mk, williams n, taussig r, wei s, roth m, hughes dt, huntley jf, fina mw, falck jr, sperandio v. targeting qsec signaling and virulence for antibiotic development. science. 2008 aug 22;321(5892):1078-80.