[Comparison of inhibitory effect of rifalazil and rifampicin against Mycobacterium ulcerans infection induced in mice].
OBJECTIVE
Buruli ulcer is a human skin disease caused by Mycobacterium ulcerans infection, which is characterized by massive skin ulceration and persistent necrotic change. In recent years Buruli ulcer has rapidly emerged as an increasingly important cause of human morbidity around the world. The disease is endemic at least 32 countries in Africa, Western Pacific, Asia and South America, and it is considered the third most common mycobacterial infection of humans after tuberculosis and leprosy. An effective chemotherapeutic regimen against Buruli ulcer disease has not been established to date. In this study, the inhibitory effect of rifalazil (RLZ) against M. ulcerans was assessed in experimentally infected mice and compared to that of rifampicin (RFP).


METHODS
Five-week-old BALB/c female mice were challenged with 25 microliters (CFU = 4 x 10(4) of M. ulcerans cultured in Middlebrook 7H9 broth in bilateral hind footpads. Mice were administered per os with a suspension of RLZ or RFP at 2.5, 5, or 10 mg/kg once daily 5 times per week starting from one day up to 6 weeks after infection. During the treatment, mice were observed weekly for footpad skin lesions and examined for footpad swelling. In addition, CFU enumeration was done on both hind footpads and spleen at 2, 4, and 6 weeks after initiating treatment.


RESULTS
In the infected control mice group, slightly erythematous lesions and moderate swelling of footpads were observed 4 weeks after the infection. Ulcerative lesion was observed 6 weeks after the infection. Mean log10 CFU/footpad (FP) was 5.22 on day 1 after the infection and increased to 5.56, 6.29, and 7.33 at 2, 4, and 6 weeks after treatment was initiated in the treated groups. On the other hand, no visible erythema, swelling or ulcerative lesion in footpads were observed in RLZ-administered groups. Furthermore, log10 CFU/FP decreased to 4.14 after only 2 weeks of initiating treatment in 2.5 mg/kg administered group, i.e. the lowest dose employed group. Log10 CFU/FP decreased to < 2.1 in 6 weeks in the 10 mg/kg administered group, which was close to the detection limit (< 1.7) of the CFU assay. By contrast, inhibitory effect on disease progression and reduction of CFU were observed only in the group of mice given 10 mg/kg among RFP-administered groups: the reduction of CFU was not observed in the early period but 6 weeks after initiating treatment.


CONCLUSIONS
These results clearly demonstrate that the in vivo anti-M.ulcerans activity of RLZ is much higher than RFP. RLZ activity against M. ulcerans can be expected to control the disease progression in the clinical applications.
Nakanaga
Kazue
K
Department of Bioregulation, Leprosy Research Center, National Institute of Infectious Diseases, 4-2-1, Aoba-cho, Higashimurayama-shi, Tokyo 189-0002, Japan. nakanaga@nih.go.jp
Saito
Hajime
H
Ishii
Norihisa
N
Goto
Masamichi
M
D000818:Animals; D000904:Antibiotics, Antitubercular / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D009165:Mycobacterium Infections, Nontuberculous / Q000188:drug therapy*; D019911:Mycobacterium ulcerans / Q000187:drug effects; D012293:Rifampin / Q000494:pharmacology / Q000627:therapeutic use*; D012294:Rifamycins / Q000494:pharmacology / Q000627:therapeutic use*; D014382:Tuberculosis, Cutaneous / Q000188:drug therapy
D000904:Antibiotics, Antitubercular; D012294:Rifamycins; C068980:KRM 1648; D012293:Rifampin