Activity of a Long-Acting Injectable Bedaquiline Formulation in a Paucibacillary Mouse Model of Latent Tuberculosis Infection

The potent antituberculosis activity and long half-life of bedaquiline make it an attractive candidate for use in long-acting/extended-release formulations for the treatment of latent tuberculosis infection (LTBI). Our objective was to evaluate a long-acting injectable (LAI) bedaquiline formulation in a validated paucibacillary mouse model of LTBI.

. M. tuberculosis lung CFU counts during 12 weeks of treatment. (Supplement Page 2)  Figure S1. M. tuberculosis lung CFU counts during 12 weeks of treatment. Lung CFU data are presented for mice receiving (A) the control regimens, (B) the daily oral bedaquiline regimens, (C) the BLAI-160 regimens, and total bedaquiline doses of (D) 160 mg/kg, (E) 320 mg/kg, and (F) 480 mg/kg. Data points represent mean values, and error bars represent standard deviation (n = 5 mice per group per time point) provided in Table 3. See Table 2 for a description of the regimens. See Tables S2-S7 for the raw CFU data at each time point. Table S1. CFU count data from quantitative cultures of the bacterial suspensions used for aerosol infections. Serial ten-fold dilutions of the bacterial suspensions used for aerosol infections were plated on non-selective 7H11 agar (0.5 mL per agar plate). The shaded cells were used to calculate CFU/mL. The lower limits of detection were 4.30 log10 CFU/mL and 1.32 log10 CFU/mL for the M. bovis rBCG30 and M. tuberculosis H37Rv infections, respectively. ++ indicates non-uniform confluent growth. + indicates individual colonies too numerous to accurately count or estimate.
---indicates not determined. Suspensions used for aerosol infections CFU counts for the following 10-fold dilutions: CFU/mL log 10 CFU/mL Table S2. CFU count data used to calculate the number of bacteria implanted in mouse lungs following aerosol infections. Mice were sacrificed the day after infection. Undiluted lung homogenates and serial ten-fold dilutions were cultured (0.5 mL per agar plate) on selective 7H11 agar ("plain" agar) and selective 7H11 agar supplemented with 40 µg/mL HYG or 4 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample.
LLOD: lower limit of detection (log10 CFU/lung). + indicates individual colonies too numerous to accurately count or estimate. ~ indicates that a precise CFU count could not be determined due to merged/touching colonies.
---indicates not determined.   Table S3. CFU count data from quantitative cultures of mouse lung homogenates at Day 0. Mouse lungs were homogenized in 2.5 mL PBS. Undiluted lung homogenate and serial ten-fold dilutions were cultured (0.5 mL per agar plate) on selective 7H11 agar ("plain" agar) and selective 7H11 agar supplemented with 40 µg/mL HYG or 4 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample, and the lower limit of detection was 0.78 log10 CFU/lung for each sample. ++ indicates non-uniform confluent growth. + indicates individual colonies too numerous to accurately count or estimate.  Table S4. CFU count data from quantitative cultures of mouse lung homogenates at Week 4. Mouse lungs were homogenized in 3.0 mL PBS. Undiluted homogenates and up to three ten-fold serial dilutions of homogenates were plated (0.5 mL per agar plate) on selective 7H11 agar with charcoal ("Plain" agar) and selective 7H11 charcoal agar supplemented with 40 µg/mL HYG or 200 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample.
Continued on next page.   Table S5. CFU count data from quantitative cultures of mouse lung homogenates at Week 8. Mouse lungs were homogenized in 3.0 mL PBS. Undiluted homogenates and up to three ten-fold serial dilutions of homogenates were plated (0.5 mL per agar plate) on selective 7H11 agar with charcoal ("Plain" agar) and selective 7H11 charcoal agar supplemented with 40 µg/mL HYG or 200 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample.
Continued on next page.       Table S6. CFU count data from quantitative cultures of mouse lung homogenates from untreated mice at Week 12. Mouse lungs were homogenized in 3.0 mL PBS. Four ten-fold serial dilutions of lung homogenates were plated (0.5 mL per agar plate) on selective 7H11 agar with charcoal ("Plain" agar) and selective 7H11 charcoal agar supplemented with 40 µg/mL HYG or 200 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample.
LLOD: lower limit of detection (log10 CFU/lung). Contam. indicates bacterial or fungal contamination; this plate was not used to calculate CFU/lung. ++ indicates non-uniform confluent growth. + indicates individual colonies too numerous to accurately count or estimate.  Table S7. CFU count data from quantitative cultures of mouse lung homogenates at Week 12. Mouse lungs were homogenized in 3.0 mL PBS. Undiluted homogenates and up to two ten-fold serial dilutions of homogenates were plated (0.5 mL per agar plate) on selective 7H11 agar with charcoal ("Plain" agar) and selective 7H11 charcoal agar supplemented with 40 µg/mL HYG or 200 µg/mL TCH. The shaded cells were used to calculate CFU/lung for each sample.