Rifampin rif

Compounds 2178, 2504, and 2150 were initially provided by Sanofi (Paris, France) for screening and initial experiments. The compounds were then resynthesized and purified at the University of North Carolina, Chapel Hill. Sources of other compounds are listed in Text S1 in the supplemental material. SAR1, TMC207, and cyclohexyl-griselimycin were provided by Sanofi (Paris, France). 8-Hydroxyquinoline (8-HQ), activated charcoal, sodium resazurin, hexaammine cobalt chloride (HexaCo), isoniazid, and rifampin (Rif) were from Sigma-Aldrich, USA, and moxifloxacin hydrochloride (Moxi) was from Santa Cruz Biotechnology, USA.

A Cornell-like model of paucibacillary TB that was previously described in the C57BL/6J mouse (Radaeva et al., 2005 (link)) was reproduced in HIS mice. Following development of clinical signs of active TB at 8 and 4 weeks post-infection in study 1 and 2, respectively, TB chemotherapy was initiated for 8 weeks. Treatment consisted of daily oral gavage for seven days a week with 750 ug of both rifampin (RIF) and isoniazid (INH) purchased from Sigma-Aldrich and dissolved in 100 μl of sterile water. Stock solutions of RIF were dissolved in a low volume DMSO and final concentration of DMSO in the delivered dose was <0.01%. Chemotherapy was provided to all treatment groups (including non-infected and HIV-infected groups) to control for drug and carrier effects and potential carryover of weak antiviral activity that is described for RIF (Clark, 1971 (link); Moshkowitz et al., 1971 (link)). Upon development of clinical signs (e.g., weight loss) following 4–8 weeks of Mtb infection and upon completion of 8 weeks of TB chemotherapy, the bacterial burden was determined in randomly selected animals using CFU enumeration of disrupted tissue. Tissue specimens were also preserved for histopathology and histochemistry analysis.

Mycobacterium smegmatis mc² 155 was grown in Middlebrook 7H9 broth (catalog number 271310; Difco, Detroit, MI) enriched with 10% (vol/vol) oleic acid-albumin-dextrose-catalase (OADC) (catalog number 212240; Becton, Dickinson, Franklin Lakes, NJ) and 0.5% (vol/vol) glycerol (catalog number G-5516; Sigma-Aldrich) in 50-ml Falcon tubes at 37°C without shaking. A concentration of 0.025% (vol/vol) tyloxapol surfactant (catalog number T8761; Sigma-Aldrich) was added as one of the treatments. Rifampin (RIF; catalog number R3501) and capreomycin sulfate (CAP) from Streptomyces capreolus (catalog number C4142) were from Sigma-Aldrich. D-AMPs D-LAK120-A (KKLALALAKKWLALAKKLALALAKK-NH₂) and D-LAK120-HP13 (KKALAHALKKWLPALKKLAHALAKK-NH₂) were supplied by China Peptides Co., Ltd. (Shanghai, China). The purity of synthesized peptides was above 80%, and the peptides were used as supplied. Both peptides were amidated at the C terminus.