micafungin in vitro 3 3 micafungin Candida Aspergillus Candida albicans fluconazole Candida tropicalis, Candida glabrata, Candida krusei Aspergillus 90 MIC90 0. g ml amphotericin B fluconazole itraconazole Candida parapsilosis Candida guilliermondii MIC90 g ml Candida Aspergillus fumigatus Cryptococcus neoformans, Trichosporon Fusarium solani, Pseudallescheria boydii Candida A. fumigatus MIC ph MIC C. albicans Key words: micafungin in vitro Immunocompromised host 3 Immunocompromised host Candida Aspergillus Cryptococcus amphotericin B azole fluorocytosine FC Azole azole Candida albicans Candida 9 0 echinocandin anidulafungin LY 3033 pneumocandin caspofungin MK099 MIC MIC NCCLS MIC 9 Anidulafungin caspofungin MIC Candida Aspergillus 9 Micafungin Coleophoma empetri echinocandin FR 9039 0 3 Dglucan in vitro
Micafungin in vitro in vitro NCCLS I Micafungin amphotericin B ; Fungizone! fluconazole ; Diflucan! 0. itraconazole ; Itrizole! Cap.0 fluorocytosine FC; flucytosine C. albicans ATCC American Type Culture Collection IFM TIMM 3 FC 00 mg 00 ml dimethyl sulfoxide MIC MIC National Committee for Clinical Laboratory Standards NCCLS M A MIC RPMI 0 3 Nmorpholino propanesulfonic acid MOPS 0. M mol L ph.0 RPMI MOPS Sporothrix schenckii RPMI MOPS RPMI MOPS 9 00 L 3 Sabouraud dextrose agar SDA; glucose Bacto peptone. Bacto agar 3 Potato dextrose agar PDA 30 Dextrose Brain heart infusion agar BHIAS. schenckii 3 CO 3 Penicillium marneffei S. schenckii PDA 30 0 Coccidioides immitis Dextrose BHIA 3 00 L.0. 0 3 cells ml C. immitis.0 0 3.0 0 cells ml Candida Saccharomyces cerevisiae, Cryptococcus neoformans, Trichosporon Aspergillus Pseudallescheria boydii 3 F. solani 30 3 C. immitis 3Sprothrix schenckii 3 CO MIC NCCLS M A 0 MIC MIC 0 MIC MIC Candida parapsilosis ATCC 09 Candida krusei ATCC C. albicans ATCC 900 MIC NCCLS M A. MIC C. albicans ATCC 900 C. parapsilosis ATCC 09 C. krusei ATCC Aspergillus fumigatus TIMM 003 MIC
Table Antifungal spectrum of micafungin Organism MIC Candida albicans ATCC 900 Candida albicans FP 33 Candida tropicalis TIMM 033 Candida glabrata ATCC 90030 Candida kefyr ATCC 3 Candida krusei ATCC Candida guilliermondii 3003 Candida parapsilosis ATCC 09 Candida stellatoidea IFM 9 Saccharomyces cerevisiae ATCC 93 Cryptococcus neoformans TIMM 03 Trichosporon cutaneum IFM 00 Trichosporon asahii TIMM 3 3 0 0 0 00 0 0 Aspergillus fumigatus TIMM 003 Aspergillus niger ATCC Aspergillus nidulans IFM 39 Aspergillus flavus ATCC 93 Aspergillus terreus IFM 0 Aspergillus versicolor IFM 0 Fusarium solani IFM 3 Pseudallescheria boydii IFM 0 00 0 00 0 00 3 3 0 0 0 0 Cladosporium trichoides IFM Exophiala dermatitidis IFM Exophiala spinifera ATCC Fonsecaea pedrosoi ATCC 3 0 00 0 0 Absidia corymbifera IFM 0 Cunninghamella elegans IFO Rhizopus oryzae IFM 0 Rhizopus microsporus var. rhizopodiformis IFM Medium: RPMI 0 mm MOPS ph 0 Inoculum: 0 to 0 3 cells ml Culture: 3 30 days 3days more than days : micafungin, : fluconazole, : itraconazole, : amphotericin B.0 0.0 0 3.0 0.0 0 cells ml 00 L ph RPMI 0 MOPS mol L mol L ph 3 30 0 30 90 RPMI MOPS HSA RPMI MOPS 0..0 MFC MIC 00 L SDA 3 99 MFC MFC Candida A. fumigatus RPMI MOPS ml C. albicans FP 33 C. albicans ATCC 900 C. glabrata ATCC 90030 C. krusei ATCC C. parapsilosis ATCC 09 C. tropicalis TIMM 033 SDA 3 RPMI MOPS.0 0.0 0 cells ml 3 ml RPMI MOPS control3
Micafungin in vitro Table MICs of micafungin against clinical isolates of yeasts Organism no of isolates Compound MIC range MIC0 b MIC90 b C. albicans 0 00 0 0 0 00 0 00 0 0 0 0 C. albicans resistant C. tropicalis 0 0 0 00 0 0 0 0 0 0 C. glabrata 3 0 0 3 C. krusei 3 3 C. parapsilosis C. guilliermondii 9 C. neoformans 0 T. cutaneum Medium: RPMI 0 mm MOPS ph 0 Inoculum: 0 to 0 3 cells ml Culture: 3 days 3days todays b MIC0 or MIC90:TheMICsat which 0 or90 of isolates are inhibited, respectively : micafungin, : fluconazole, : itraconazole, : amphotericin B 3 9 RPMI MOPS C. albicans ATCC 900 SDA 30 RPMI MOPS.0 0.0 0 cells ml ml ml MIC RPMI MOPS 0 3.0 0 cells ml RPMI MOPS ml ml 3 II AMPH
Table 3 MICs of micafungin against clinical isolates of Aspergillus species Organism no of isolates Compound MIC range MIC0 b MIC90 b A. fumigatus 39 0 00 0 0 A. niger 0 00 0 0 A. flavus 0 00 0 0 0 0 A. terreus 0 0039 0 0 0 00 Medium: RPMI 0 mm MOPS ph 0 Inoculum: 0 0 3 cells ml Culture: 33 days b MIC0 or MIC90:TheMICsat which 0 or90 of isolates are inhibited, respectively : micafungin, : fluconazole, : itraconazole, : amphotericin B B Table Candida Saccharomyces cerevisiae, Aspergillus C. neoformans, Trichosporon F. solani, P. boydii MIC Candida C. neoformans Trichosporon cutaneum MIC Table C. albicans C. tropicalis, C. glabrata C. krusei 90 MIC90 0. ml C. parapsilosis C. guilliermondii MIC90 ml C. neoformans T. cutaneum MIC ml Aspergillus Aspergillus MIC Table 3 Aspergillus MIC90 0.033 ml 3 MIC Table Exophiala spinifera, Fonsecaea pedrosoi Cladosporium trichoides Table MICs of micafungin against dematiaceous fungi Organism MIC range Compound no of isolates E. dermatitidis E. spinifera F. pedrosoi C. trichoides 0 00 0 0 Medium: RPMI 0 mm MOPS ph 0 Inoculum: 0 0 3 cells ml Culture: 33 todays to0days : micafungin, : fluconazole, : itraconazole, : amphotericin B MIC 0. ml Exophiala dermatitidis 3 MIC ml 3 ml
Micafungin in vitro Table MICs of micafungin against dimorphic fungi Organism no of isolates: yeastlike form mycelial form Compound Yeastlike form MIC range Mycelial form H. capsulatum 3 3 0 00 0 0 0 0 B. dermatitidis 3 0 0039 0 00 0 00 3 P. brasiliensis 0 0039 0 0039 0 00 0 00 0 0 P. marneffei 0 00 0 0 S. schenckii C. immitis b b b b 0 0 0 0 Medium: RPMI 0 mm MOPS ph 0 Inoculum: Yeastlike form, 0 to 0 0 P. marneffei 0 0 3, S. schenckii 0 0 cells ml Mycelial form, 0 to 0 0 P. brasiliensis 0 0, C. immitis 0 0 3 cells ml Culture: Yeastlike form 3 todays,aerobic culture S. schenckii,3 CO,withshaking Mycelial form, C. immitis,3 3 todays,aerobic culture b :Nottested : micafungin, : fluconazole, : itraconazole, : amphotericin B MIC Table H. capsulatum, B. dermatitidis, P. brasiliensis, P. marneffei S. schenckii H. capsulatum, B. dermatitidis, S. schenckii C. immitis MIC 0.00 ml AMPH B P. marneffei MIC 0.033 ml AMPH B P. brasiliensis MIC ml 3. MIC ph HSA MIC Table Candida A. fumigatus MIC ph HSA MIC MFC Candida A. fumigatus MFC Table C. albicans C. albicans C. glabrata C. krusei MFC90 0. 0. 0.033 0. ml C. tropicalis C. guilliermondii MFC90 ml C. parapsilosis
Table Influence of culture conditions on MIC MIC Culture condition C. albicans ATCC 900 C. parapsilosis ATCC 09 C. krusei ATCC A. fumigatus TIMM 003 ph b 0 0 0 00 0 0039 0 00 Inoculum size c cells ml 0 0 3 0 0 0 0 0 0 0 0039 0 00 0 00 Addition of human serum d 0 0 30 90 3 0 00 Addition of HSA d 0 0 0 0 Culture : 3 days 3days b RPMI 0 mm MOPS was adjusted to a ph of,, and with mol L HClormol L NaOH,Inoculum size was 0 0 3 cells ml c RPMI 0 mm MOPS ph 0 was used asatestmedium d RPMI 0 mm MOPS ph 0 was used as a test medium, inoculum size was 0 0 3 cells ml MFC90 ml Candida MFC90 ml ml A. fumigatus MFC ml C. albicans FP 33 Fig 0.00 ml 99 3 ml 3 C. albicans C. glabrata, C. krusei, C. parapsilosis C. tropicalis Fig C. albicans C. glabrata C. krusei MIC 99C. parapsilosis C. tropicalis Candida MIC 3 9 Changes of viable counts log CFU 0 3.03 Detection limit 0.0039 0.0 0.0 0. Drug concentrationg/ml : micafungin, : amphotericin B, : fluconazole : itraconazole Fig. Fungicidal activity against Candida albicans FP 33 after hours exposure. log CFU: logarithm of CFU after hours exposure logarithm of CFU at time zero C. albicans ATCC 900 MIC Fig 3 C. albicans FC 3
Micafungin in vitro Table MFCs of micafungin for clinical isolates of Candida species and Asperigillus fumigatus Organism no of isolates Compound MFC range MFC0 b MFC90 b C. albicans C. albicans resistant C. tropicalis 0 0 0 0 C. glabrata C. krusei 0 C. parapsilosis 0 C. guilliermondii 0 A. fumigatus MFC: Minimum fungicidal concentration more than 99 of the original inoculum was killed Medium: RPMI 0 mm MOPS ph 0 Inoculum: 0 to 0 3 cells ml b MFC0 or MFC90:TheMFCsat which 0 or90 of isolates are inhibited, respectively : micafungin, : fluconazole, : itraconazole, : amphotericin B III in vitro NCCLS M A Candida MIC MIC M A 0 AMPH B MIC Candida in vitro C. albicans, C. tropicalis, C. glabrata C. krusei MIC90 C. albicans FC 3 FC C. parapsilosis C. guilliermondii MIC Candida 9 C. albicans FP 33 ATCC 900
C. albicans FP33 C. albicans ATCC900 Viable counts Log of cfu/ml 3 Viable counts Log of cfu/ml 3 0 3 9 0 3 9 Time h 0 3 9 0 3 9 Time h C. glabrata ATCC90030 C. krusei ATCC Viable counts Log of cfu/ml 3 Viable counts Log of cfu/ml 3 0 3 9 0 3 9 Time h 0 3 9 0 3 9 Time h C. parapsilosis ATCC09 C. tropicalis TIMM033 Viable counts Log of cfu/ml 3 Viable counts Log of cfu/ml 3 0 3 9 0 3 9 Time h 0 3 9 0 3 9 Time h : micafungin, : amphotericin B Fig. Timekill curve against Candida albicans FP 33 C. albicans ATCC 900 Candida glabrata ATCC 90030 Candida krusei ATCC Candida parapsilosis ATCC 09 and Candida tropicalis TIMM 033. ConcentrationsControl MIC MIC MICMICMIC C. albicans MFC90 C. albicans Candida MFC Candida MIC
Micafungin in vitro MICg/mL > 0. 0.0 0.0 0 3 9 0 3 Number of passage times -FC : micafungin, : amphotericin B, : fluconazole : itraconazole Fig 3. Change in the susceptibility of Candida albicans ATCC 900 to micafungin after repeated exposure to subinhibitory concentrations of. MIC A. fumigatus in vivo 3 3 A. fumigatus MIC Aspergillus MIC in vivo MIC M A Aspergillus MIC90 MIC C. neoformans, T. cutaneum, T. asahii, F. solani, P. boydii Aspergillus MIC caspofungin anidulafungin 9 H. capsulatum B. dermatitidis 3 C. immitis MIC H. capsulatum B. dermatitidis MIC P. brasiliensis MIC ml MIC ml P. marneffei S. schenckii MIC H. capsulatum B. dermatitidis 3 Dglucan FKS C. neoformans FKS C. neoformans 3 Dglucan 3 Dglucan chitin chitosan 3 Dglucan 3 Dglucan F. solani, P. boydii B. dermatitidis P. brasiliensis glucan glucan 3 3 33 in vivo MIC MIC MIC 3 Candida Aspergillus Anaissie E: Opportunistic mycoses in the immunocompromised host: experience at a cancer center and review. Clin Infect Dis : S 3 S3 99 Hadley S Karchmer A W: Fungal infections in solid
organ transplant recipients. Infect Dis Clin N Am 9: 0 0 99 3 Dixon D M McNeil M M Cohen M L et al.: Fungal infections: a growing threat. Public Health Re : 3 99 Morrison V A Haake R J Weisdorf D J: The spectrum of non Candida fungal infections following bone marrow transplantation. Medicine : 9 993 Padhye A A Hampton A A Hampton M T et al.: Chromoblastomycosis caused by Exophiala spinifer Clin Infect Dis : 33 33 99 Patterson T F Andriole V T Zervos M J et al.: The epidemiology of pseudallescheriasis complicating transplantation : nosocomial and community acquired infection. Mycoses 33: 9 30 990 Singh N Chang F Y Gayowski T et al.: Infections due to dematiaceous fungi in organ transplant recipients: case report and review. Clin Infect Dis : 39 3 99 Thomas A H: Suggested mechanisms for the antimycotic activity of the polyene antibiotics and the N substituted imidazoles. J Antimicrob Chemother : 9 9 9 9 Hitchcock C A Pye G W Johnson E M et al.: Fluconazole resistance in Candida glabrata. Antimicrob Agents Chemother 3: 9 9 993 0 Debono M Gordee R S: Antibiotics that inhibit fungal cell wall development. Annu Rev Microbiol : 9 99 Vazquez J A Lynch M Sobel J D: In vitro activity of a new pneumocandin antifungal agent L 33 0 against azolesusceptible and resistant Candida and Torulopsis species. Antimicrob Agents Chemother 39: 9 9 99 Verweij P E Oakley K L Morrissey J et al.: Efficacy of LY 3033 against amphotericin B susceptible and resistant Aspergillus fumigatus in amurine model of invasive aspergillosis. Antimicrob Agents Chemother : 3 99 3 Petraitis V Petraitiene R Groll A H et al.: Antifungal efficacy safety and singledose pharmacokinetics of LY 3033 a novel echinocandin B in experimental pulmonary aspergillosis in persistently neutropenic rabbits. Antimicrob Agents Chemother : 9 90 99 Bartizal K Gill C J Abruzzo G K et al.: In vitro preclinical evaluation studies with the echinocandin antifungal MK099 L3 Antimicrob Agents Chemother : 3 33 99 National Committee for Clinical Laboratory Standards: Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard. NCCLS document M A. National Committee for Clinical Laboratory Standards Wayne Pa., 99 Krishnarao T V Galgiani J N: Comparison of the in vitro activities of the echinocandin LY 3033 the pneumocandin MK099 and fluconazole against Candida Species and Cryptococcus neoformans Antimicrob Agents Chemother : 9 90 99 Ingroff A E: Comparison of in vitro activities of the new triazole SCH 9 and the echinocandins MK 099 L3 and LY 3033 against opportunistic filamentous and dimorphic fungi and yeasts. J Clin Microbiol 3: 90 9 99 Poeta M D Schell W A Perfect J R: In vitro antifungal activity of pneumocandin L3 against a variety of clinically important molds. Antimicrob Agents Chemother : 3 3 99 9 Zhanel G G Karlowsky J A Harding G A J et al.: In vitro activity of a new semisynthetic echinocandin LY3033 against systemic isolates of Candida species Cryptococcus neoformans, Blastomyces dermatitidis,and Aspergillusspecies. Antimicrob Agents Chemother : 3 99 0 Iwamoto T Fujie A Sakamoto K et al.: WF 99 A B and C novel antifungal lipopeptides I. Taxonomy fermentation isolation and physico chemical properties. J Antibiotics : 0 09 99 Tomishima M Ohki H Yamada A et al.: FK 3 a novel watersoluble echinocandin lipopeptide: synthesis and antifungal activity. J Antibiotics : 999 : Micafungin Candida albicans Aspergillus fumigatus 0 S : 0 9 00 3 Abruzzo G K Flattery A M Gill C J et al.: Evaluation of the echinocandin antifungal MK099 L3 : efficacies in mouse models of disseminated aspergillosis candidiasis and cryptococcosis. Antimicrob Agents Chemother : 333 33 99 Kurtz M B Bernard E M Edwards F F et al.: Aerosol and parenteral pneumocandins are effective in a rat model of pulmonary aspergillosis. Antimicrob Agents Chemothe. 39: 9 99 Oakley K L Moore C B Denning D W: In vitro activity of the echinocandin antifungal agent LY 303 3 in comparison with itraconazole and amphotericin B against Aspergillus spp. Antimicrob Agents Chemother : 30 99 Thompson J R Douglas C M Li W et al.: A glucan synthase FKS Homolog in Cryptococcus neoformans is single copy and encodes an essential function. J Bacteriol : 3 999 SietsmaJH Wessels J G H: Apical wall biogenesis. The Mycota I Growth Differentiation and sexuality Wessels J G H Meinhardt F edsp. SpringerVerlag Berlin 99 Domer J E: Monosaccharide and chitin content of cell walls of Histoplasma capsulatum and Blastomyces dermatitidis JBacteriol 0: 0 9 9 Kanetsuna F Carbonell L M: Cell wall glucans of the yeast and mycelial forms of Paracoccidioides brasiliensis JBacteriol 0: 0 90 30 Kanetsuna F Carbonell L M: Cell wall composition of the yeastlike and mycelial forms of Blastomyces
Micafungin in vitro dermatitidis JBacteriol 0: 9 9, 9 3 Kanetsuna F Carmonell L M Azuma I et al.: Biochemical studies on the thermal dimorphism of Paracoccidioides brasiliensis J Bacteriol 0: 0 9 3 Maresca B Kobayashi G S: Dimorphism in Histoplasma capsulatum: amodelforthestudyofcell differentiation in pathogenic fungi. Microbiol Rev 3: 09 99 33 SanBlas G: The cell wall of fungal human pathogens: its possible role in hostparasite relationships. Mycopathologia 9: 9 9 3 : micafungin in vitro 0 S : 9 00 In vitro activity of a new lipopeptide antifungal agent micafungin against a variety of clinically important fungi Fumiaki Ikeda Kazumi Otomo Tohru Nakai Yoshihiko Morishita Katsuyuki Maki Shuichi Tawara Seitaro Mutoh Fumio Matsumoto and Shogo Kuwahara 3 Medicinal Biology Research Laboratories Fujisawa Pharmaceutical Co., Ltd., KashimYodogawaku Osaka 3 Japan Kanagawa Prefectural Nursing and Hygienic School Hospital 3 Toho University School of Medicine The in vitro antifungal activity and spectrum of micafungin were compared with those of amphotericinb fluconazole and itraconazole using a broth microdilution method as specified by the National Committee for Clinical Laboratory Standards NCCLS document M A. exhibited broadspectrum activity against clinically important pathogens including Candida species and Aspergillus species and its MIC90 levels against C. albicans including resistant C. albicansc. tropicalis, C. glabrata, C. krusei and Aspergillus species were 0. ml which were lower than those for the other antifungal agents tested. The MIC90 levels of against C. parapsilosis and C. guilliermondii were and ml respectively which were comparable to orhigher than those for the other antifungal agents tested. exhibited concentrationindependent fungicidal activity at concentrations higher than the MIC against most Candida species. In contrast the MFCs of against A. fumigatus isolates were much higher than the MICs of the other agents indicating that its action is fungistatic against this species. showed moderate to weak activity against most dematiaceous fungi and had no activity against Cryptococcus neoformans, Trichosporon species Fusarium solani, Pseudallesheria boydii and zygomycetes. Although showed potent activity against the mycelial form of dimorphic fungi it had weak or no activity against their yeastlike form. Neither the ph of the test medium nor the inoculum size greatly affected the MIC values of while addition of human serum or human serum albumin increased the MIC values against Candida species and A. fumigatus In experiments on resistance induction the MIC of for C. albicans was not significantly changed; indicating that there is a low probability of induced resistance.