AAC Revised. Activity of a Novel Cyclic Lipopeptide, CB-183,315 Against Resistant Clostridium difficile
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1 AAC Accepts, published online ahead of print on 5 March 2012 Antimicrob. Agents Chemother. doi: /aac Copyright 2012, American Society for Microbiology. All Rights Reserved. 1 AAC Revised Activity of a Novel Cyclic Lipopeptide, CB-183,315 Against Resistant Clostridium difficile and Other Gram Positive Aerobic and Anaerobic Intestinal Pathogens D. R. SNYDMAN 1,2, N. V. JACOBUS 1, L. A. MCDERMOTT 1 1 Tufts Medical Center and 2 Tufts University School of Medicine, Boston, MA, USA We evaluated the activity of CB-183,315 against C. difficile including strains resistant to fluoroquinolones, metronidazole and with elevated MICs to vancomycin as well as other Gram positive intestinal pathogens. The MICs of CB-183,315 against all C. difficile isolates were < 1 μg/ml. CB-183,315 had greater activity than vancomycin and metronidazole against C. difficile and was more active than the comparators against VRE. CB-183,315 also had excellent activity against MRSA, other Clostridium spp. and Peptostreptococcus spp. 1
2 In the past decade the incidence and severity of Clostridium difficile associated diarrhea has increased significantly. Outbreaks caused by a highly virulent and epidemic strain, (BI/NAP1/027) have emerged in Canada, United States, Europe and Asia. [4,10,12,13,16,21] There has been little incentive to perform susceptibility testing on C. difficile isolates since these isolates have been thought to be uniformly susceptible to metronidazole and vancomycin, the primary antimicrobials used in treatment. Recently, isolates with elevated MICs to vancomycin and metronidazole have been reported. [1, 17, 19, 20] Knowing that the rate of response to C. difficile treatment is less than optimal, and with knowledge that resistance to the primary agents may be occurring, there is a need to develop new agents for the treatment of Clostridium difficile infections (CDI). [2, 9,11,16] CB-183,315 is a novel, cyclic lipopeptide analogue of daptomycin with very good activity against anaerobic-gram-positive bacteria, including C. difficile, and limited activity against Gram-negative pathogens. [3,20] A recent phase 2 clinical trial evaluating the safety and effectiveness of CB-183,315 administered orally in doses of 125 and 250 mg bid concluded that either dose was safe and well tolerated. The authors also concluded that CB-183,315 sustained better cure rates than vancomycin in the treatment of CDI. [18] We undertook this study to determine the activity of CB-183,315 against isolates of C. difficile resistant to metronidazole, with elevated MICs to vancomycin and highly resistant to fluoroquinolones and clindamycin. Other intestinal pathogens, such as Enterococcus spp. (VSE and VRE), Staphylococcus aureus (MSSA and MRSA), Peptostreptococcus spp. and Clostridium spp. were also included. The antimicrobials were obtained from their respective manufacturers or distributor. 2
3 Standard powders were diluted according to specifications. Resistance percentages were calculated using breakpoints recommended by Clinical and Laboratory Standards Institute (CLSI). For agents without recommendations for anaerobic bacteria, such as vancomycin, minocycline and linezolid, the CLSI breakpoint for resistance for aerobes for systemic infections was used. [5,6,7] CB-183,315 is an investigational compound, therefore resistance breakpoints were not calculated. The MICs of the antibiotics against 369 isolates were determined by agar dilution following CLSI, M11-A-7 and CLSI, M7-A8 recommendations. [5,6] The CLSI indicated ATCC reference control strains were included in each test. The Ca++ concentration in the media (Brucella and Mueller Hinton agars) was adjusted to 50 mg/l as recommended by the manufacturer. Moxifloxacin was used to determine susceptibility to the fluoroquinolones. Table 1 shows the susceptibilities of 55 C. difficile isolates to CB-183,315 compared with those of the other agents. It should be noted that many multi-drug resistant isolates were included in this study, 38.2% were resistant to moxifloxacin and 18.2% were resistant to clindamycin. In addition 23.6% showed elevated MICs to metronidazole (MIC > 8 µg/ml) and 21.8% had MICs for vancomycin of > 4 µg/ml. In order to underscore the activity of CB- 183,315 the results are shown in subsets of specific susceptibilities. Among the agents, CB-183,315 was the most active against all C. difficile isolates, MIC90 of < 0.25 µg/ml. All C. difficile isolates were inhibited at concentrations of < 1 µg/ml. CB- 183,315 was more active than the two agents routinely used for treatment of CDI, namely vancomycin and metronidazole. CB-183,315 exceeded the potency of vancomycin by 4 fold and that of metronidazole by greater than 16-fold. The activity of CB-183,315 to C. difficile was identical for moxifloxacin-resistant isolates 3
4 as well for the moxifloxacin-susceptible. Activity was maintained for those isolates that had elevated MIC s to metronidazole as well as those with decreased susceptibility to vancomycin. The activity of CB-183,315 and the comparative agents against other Gram-positive isolates are illustrated in Table 2. All enterococcal isolates were inhibited by CB-183,315 at concentrations of < 2 µg/ml. All S. aureus isolates (MRSA and MSSA) were inhibited at concentrations of CB-183,315 of < 1 µg/ml. The MIC range of CB-183,315 against 33 Clostridium spp. was µg/ml. 70% of Clostridium spp. were inhibited by < 1 µg/ml of CB-183,315. Two strains, one C. clostridioforme and a Clostridium spp., showed MICs of 4 µg/ml. CB-183,315 inhibited 93% of Peptostreptococcus spp. at 0.5 µg/ml. Among 45 Peptostreptococcus spp, 2 isolates showed CB-183,315 MICs of 1 µg/ml and another MIC of 2 µg/ml. Eighty six B. fragilis group isolates demonstrated MIC s > 16 µg/ml, thus confirming the limited activity of CB-183,315 against anaerobic gram-negative bacteria. CB-183,315 is structurally related to daptomycin and appears to share a common mechanism of action, i.e., membrane depolarization and loss of cell viability.[14] In vitro studies by Citron et al demonstrated bactericidal activity. [3] In addition, development of resistance to CB-183,315 is a rare event as shown by in vitro serial passage.[14] Besides these in vitro findings, a hamster model of CDAD demonstrated potent efficacy, similar to that of vancomycin, against initial onset of the disease. [15] To date, results of a clinical trial, published in abstract form, demonstrated that CB-183,315 was efficacious and well tolerated for the treatment of CDI in adults.[18] Our results and those of Citron et al [3] confirm the excellent activity of CB-183,315 against C. difficile, and its enhanced activity against C. difficile resistant to fluoroquinolones, 4
5 clindamycin, metronidazole and with elevated MICs to vancomycin. These data taken together with its reported bactericidal activity, rare resistance development, effectiveness in an animal 109 model and in clinical trials constitute an excellent pre-clinical profile placing CB-183,315 as a prominent agent for clinical development in the treatment of CDI particularly in infections caused by resistant strains. [3, 14, 15, 18] Part of this study was presented at the 50th Interscience Conference on Antimicrobial Agents and Chemotherapy, 2010 and at the European Congress on Microbiology and Infectious Diseases, This study was sponsored by a grant from Cubist Pharmaceuticals, Lexington, MA. Downloaded from on November 6, 2018 by guest 5
6 References: 1. Baines, S.D., R. O Connor, J. Freeman, W. N. Fawley, C. Harmanus, P. Mastrantonio, E. J. Kuijper, M. H. Wilcox Emergence of reduced susceptibility to metronidazole in J. Antimicrob. Chemother. 62: Citron, D. M., F. Barakhani, E. J. C. Goldstein, K. Nagaro, S. Sambol, P. Sears, Y-K. Shue Typing and susceptibility of bacterial isolates from the fidaxomicin (OPT-80) phase II study for C. difficile infection. Anaerobe. 15: Citron, D. M., K. L. Tyrell, C. V. Merriam and E. J. C. Goldstein In vitro activity of CB- 183,315, vancomycin and metronidazole against 556 strains of Clostridium difficile, 445 other intestinal anaerobes and 56 Enterobacteriaceae species Antimicrob. Agents Chemother. published ahead of print 19 December 2011, doi: /aac Clements, A. C., R. J. Magalhaes, A. J. Tatem, D. L. Paterson, and T. V. Riley Clostridium difficile PCR ribotype 027: assessing the risks of further worldwide spread. Lancet Infect. Dis. 10: Clinical and Laboratory Standards Institute Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; Approved Standard- Eighth Edition. CLSI Document M7-A8 (ISBN ). Clinical and Laboratory Standards Institute. Wayne, PA. USA. 6. Clinical and Laboratory Standards Institute Methods for antimicrobial susceptibility testing of anaerobic bacteria. Approved standard. NCCLS Document M11-A7. Clinical and Laboratory Standards Institute. Wayne, PA. 6
7 Clinical Laboratory Standards Institute CLSI Document M100-S-21. Clinical and Laboratory Standards Institute. Wayne, PA. Performance standards for antimicrobial susceptibility testing: Nineteenth informational supplement. 8. George, W. L., V. L. Sutter, D. M. Citron, and S. M. Finegold Selective and differential medium for isolation of Clostridium difficile. J. Clin. Microbiol. 9: Hecht, D. W., et al., In vitro activities of 15 antimicrobial agents against 110 toxigenic Clostridium difficile infections. Clin. Infect. Dis. 47: Hsu. M. S., J. T. Wang, W. K. Huang, Y. C. Liu and S. C. Chang Prevalence and clinical features of Clostridium difficile-associated diarrhea in a tertiary hospital in northern 11. Karlowsky, J. A., N. M. Laing, G. G. Zhanel In vitro activity of OPT-80 tested against clinical isolates of toxin-producing Clostridium difficile. Antimicrob. Agents Chemother. 52: Labbe, A.C., L. Poirier, D. MacCannell, T. Louie, M. Savoie, C. Beliveau, M. Lavardiere, J. Pepin Clostridium difficile infections in a Canadian tertiary care hospital before and during regional epidemic associated with the BI/NAP1/027 strain. Antimicrob. Agents Chemother. 52: Lin, Y-C., Y-T. Huang, P-J. Tsai, T-F. Lee, N-Y. Lee, C-H. Liao, S-Y. Lin, W-C. Ko, and P-R. Huenh Antimicrobial susceptibilities and molecular epidemiology of Clinical isolates of Clostridium difficile in Taiwan. Antimicrob. Agents Chemother. 551: Mascio, C., K. Townsend Howland and J. Silverman C Abstr.50 th Intersci. Conf. Antimicrob. Agents Chemother. American Society for Microbiology, Boston, MA. 7
8 Mortin, L.I., A.D.G. Van Praagh, S. Zhang, et al., B-707. Abstr. 50 th Intersci. Conf. Antimicrob. Agents Chemother. American Society for Microbiology, Boston, MA. 16. O Connor, J. R., M. A. Galang, S. P. Sambol, D. W. Hecht, G. Vedantam, D. N. Gerding, S. Johnson Rifampin and rifaximin resistance in clinical isolates of Clostridium difficile. Antimicrob. Agents Chemother. 52: Noren, T., I. Alriksoon, T. Akerlund, L. G. Burman and M. Unemo In vitro susceptibility to 17 antimicrobials of clinical Clostridium difficile isolates collected in in Sweden. Clin. Microbiol. Infect. 16: Patino, H., C. Stevens, T. Louie, et al K-205a. Abstr. 51 st Intersci. Conf. Antimicrob. Agents Chemother. American Society for Microbiology, Chicago, IL. 19. Pelaez, T., L. Alcala, R. Alonso, M. Rodriguex-Creixems, J. M. Garcia-Lechuz, and E. Bouza Reassessment of Clostridium difficile susceptibility to metronidazole and vancomycin. Antimicrob. Agents Chemother. 46: Snydman, D.R., N.V. Jacobus, L.M. McDermott E Abstr.50 th Intersci. Conf. Antimicrob. Agents Chemother. American Society for Microbiology, Boston, MA. 21. Taori, S. K., V. Hall, I. R. Poxton Changes in antibiotic susceptibility and ribotypes in Clostridium difficile isolates from southern Scotland, J. Med. Micro. 59:
9 Table 1 : Activities of CB-183,315 and Comparative Agents Against Clostridium difficile Isolates 1, 2 Species Antibiotic MIC Range MIC 50 MIC 90 % Resistant 3 [No. of Isolates] ( g/ml) Clostridium difficile CB-183,315 < < NA [55] Vancomycin < Pip:tazo < Amox:Clav < Minocycline < < Moxifloxacin < Meropenem < Metronidazole < Linezolid < Clindamycin < >16 2 > C. difficile (Quinolone-res.) CB-183,315 All < <0.125 <0.125 NA [21] Vancomycin Pip:tazo < Amox:Clav < Minocycline < < Moxifloxacin 8 - >16 16 > Meropenem < Metronidazole Linezolid < Clindamycin 1 - >16 16 > C. difficile (Quinolone-suscept.) CB-183,315 < < NA [34] Vancomycin < Pip:tazo < Amox:Clav < Minocycline < < Moxifloxacin < Meropenem < Metronidazole < Linezolid < Clindamycin < > C. difficile (Metronidazole MIC > 8 g/ml) CB-183,315 < < NA [13] Vancomycin Pip:tazo Amox:Clav Minocycline < Moxifloxacin < Meropenem < Metronidazole Linezolid < Clindamycin < >16 2 > C. difficile (Metronidazole MIC < 4 g/ml) CB-183,315 < < NA [42] Vancomycin Pip:tazo < Amox:Clav < Minocycline < Moxifloxacin < Meropenem < Metronidazole < Linezolid < Clindamycin < >16 2 >
10 Table 1 : Activities of CB-183,315 and Comparative Agents Against Clostridium difficile Isolates 1, 2 (Continuation) Species Antibiotic MIC Range MIC 50 MIC 90 % Resistant 3 [No. of Isolates] ( g/ml) C. difficile (Vancomycin MIC < 2 g/ml) CB-183,315 < < NA [43] Vancomycin Pip:tazo Amox:Clav Minocycline Moxifloxacin Meropenem Metronidazole Linezolid Clindamycin C. difficile (Vancomycin MIC > 4 g/ml) CB-183,315 All NA [12] Vancomycin All Pip:tazo Amox:Clav Minocycline Moxifloxacin Meropenem Metronidazole Linezolid Clindamycin MICs determined by agar dilution with the Ca++ agar concentration adjusted to 50 mg/ml. 2 Isolates classified as to susceptibility [or resistance] to moxifloxacin, metronidazole and vancomycin. 2 Resistance breakpoints are those recommended by CLSI. Resistance breakpoint for CB-183,315 have not been established, thus NA. CLSI has not issued breakpoint recommendations for the following agents: vancomycin, minocycline and linezolid, thus the breakpoints for aerobes were used.
11 Table 2: Activities of CB-183,315 and Comparative Agents Against Other Gram-Positive Intestinal Isolates 1 Species Antibiotic MIC Range MIC50 MIC90 % Resistant 2 [No. of Isolates] (mg/ml) Enterococcus spp. CB-183,315 < NA Vancomycin < > 64 2 > [60] Ampicillin > 32 1 > Minocyclyne < Levofloxacin > Erythromycin > 16 > 16 > Metronidazole > 32 > 32 > Linezolid Enterococcus spp. CB-183,315 < NA (Vancomycin-resistant, VRE) Vancomycin > 64 > 64 > [21] Ampicillin > 32 > 32 > Minocyclyne < Levofloxacin All > 8 > 8 > Erythromycin All > 8 > 8 > Metronidazole All > 32 > 32 > Linezolid Enterococcus spp. CB-183,315 < NA (Vancomycin-susceptible, VSE) Vancomycin < [39] Ampicillin > Minocyclyne < Levofloxacin > 8 2 > Erythromycin All > 8 > 8 > Metronidazole All > 32 > 32 > Linezolid Staphylococcus aureus (MSSA) CB-183, NA [12] Oxacillin Vancomycin Ampicillin 1 - > 4 2 > Amox:Clav Meropenem All < < < Minocycline All < 0.5 < 0.5 < Levofloxacin > Erythromycin 2 - > 16 > 16 > Linezolid Cefuroxime Tmp/Sulfa All < 0.5 < 0.5 < Staphylococcus aureus (MRSA) CB-183, NA [12] Oxacillin 4 - > 8 > 8 > Vancomycin All Ampicillin All > 4 > 4 > Amox:Clav 2 - > 16 8 > Meropenem > Minocycline Levofloxacin > 8 > 8 > Erythromycin All > 16 > 16 > Linezolid Cefuroxime 2 - > 64 > 64 > Tmp/Sulfa < >16 <
12 Table 2: Activities of CB-183,315 and Comparative Agents Against Other Gram-Positive Intestinal Isolates 1 (Continuation) Species Antibiotic MIC Range MIC 50 MIC 90 % Resistant 2 [No. of Isolates] (mg/ml) Clostridium spp. CB-183,315 < NA [33] Vancomycin Pip:tazo < < Amox:Clav Minocycline < < Moxifloxacin < > Meropenem < < Metronidazole Linezolid Clindamycin <0.5 - > Peptostreptococcus spp. 4 CB-183,315 < < NA [45] Vancomycin < < Pip:tazo < < 0.5 < Amox:Clav < Minocycline < < Moxifloxacin < < Meropenem < < Metronidazole < > Linezolid < Clindamycin < >16 1 > Miscellaneous Gram-positive Bacilli 5 CB-183, [6] Vancomycin < > Pip:tazo < Amox:Clav Minocycline < Moxifloxacin < Meropenem < Metronidazole > Linezolid < Clindamycin < Bacteroides fragilis group CB-183,315 > 16 > 16 > [86] Pip:tazo < Amox:Clav Minocycline < Moxifloxacin < > Meropenem < > Metronidazole < 1-2 < 1 < 1 0 Linezolid < Clindamycin < >128 1 > MICs determined by agar dilution with a Ca++ concentration for CB-183,315 was adjusted to 50 mg/l and confirmed by Laboratory Specialist Inc. Resistance breakpoints are those recommended by CLSI. Resistance breakpoint for CB-183,315 have not been established, thus NA. CLSI has not issued breakpoint recommendations for the following agents: vancomycin, minocycline and linezolid, thus the breakpoints for aerobes were used. Include: 10 C. perfringens, 5 C. cadaveris, 2 C. innocuum, 2 C. bifermentans, 2 C. histolyticum, 1 C. subterminale, 1 C. clostridioformis 1 C. ramosum, 1 C. septicum and 6 Clostridium spp. Include: 10 Finegoldia magna (P. magnus ), 5 Peptonophilus asaccharolyticus, 3 Ps. micros, 1 Ps. anaerobius, 1 Ps. tetradius, and 25 Peptostreptococcus spp. (not speciated) Include: 1 Actinomyces spp., 2 Bifidobacterium spp., 1 Lactobacillus casei, 2 Lactobacillus spp. The number of isolates too small to determine other statistics.
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