ENTERNATIONAL “W
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`JOURNAL OF
`
`timierohial
`Agents
`
`
`
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`E:5
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`_.§‘_-__..___._?._,.:'—_-_.’-H24
`
` ANTIBACTERIAL
`THERAPY
`
`1 }
`
`ANT1 VIRAL
`THEITAPY
`
`iANTIPARASITIC
`|THEI2APY
`
`'ANTIFU_NGAL
`iTHEI1APY
`
`I _
`
`|IMMUNOTHE RAPY
`
`
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`i E r T f 1
`
`T’
`
`
`inle Official Journal ofthe International Society ofChemotherapy
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`Copyright
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`l I
`
`I
`B890l4457268A
`
`Vol. 18/3
`
`INTERNATIONAL JOURNAL OF ANTIMICROBIAL AGENTS
`
`September 7001
`
`Cited in: Medline; Index Medicus; EMBASE/Excerpta Medica; Elsevier BIOBASE/Current Awareness in Biological Sciences (CABS);
`Cambridge Scientific Abstracts: CAB Abstracts; Biosis; Current Contents (Life Sciences)
`
`CONTENTS
`
`Review
`
`Role of sultamicillin and ampicillin/sulbactam in the treatment of upper and lower bacterial
`respiratory tract infections
`H. Lode
`
`Original Articles
`Antibiotic susceptibility of bacterial strains isolated from patients with community—acquired
`urinary tract infections
`R. Daza, J. Gutiérrez and G. Piédrola
`
`to fluoroquinolone antibiotics as
`In vitro susceptibilities of Bartonella and Rickettsia spp.
`determined by immunofluorescent antibody analysis of infected Vero cell monolayers
`_T.J. Ives, E.L. Marston, R.L. Regnery and J.D. Butts
`
`Antibiotic therapy of Ventilator-associated pneumoniaaa reappraisal of rationale in the era of
`bacterial resistance
`
`V. Sintcherzko, J.R. Iredell and G.L. Gilbert
`
`Influence of diminished susceptibility of Streptococcus pneumoniae to ciprofloxacin on the serum
`bactericidal activity of gernifloxacin and trovafloxacin after a single dose in healthy volunteers
`J. Prieto, L. Aguilar, F. Fuentes, M.J. Giménez, L. Alou, R. Dal-Re’, F. Sorgel and J. Frias
`
`Vancomycin‘
`administration
`
`levels
`
`in human aqueous humour
`
`after
`
`intravenous
`
`and subconjunctival
`
`M. Souli, G. Kopsinis, E. Kavouklis, L. Gabriel and H. Giamarellou
`
`Liposomes and PLG microparticles as sustained release antitubercular drug carriers—an in
`vitro~in vivo study
`M. Durt and GK. Khuller
`
`Determination of linezolid in human serum and urine by high—performance liquid chromatography
`K. Borner, E. Bomer and H. Lode.
`
`245
`
`253
`
`Contents continued nside
`
`CO
`
`.
`
`This journal is part of ContentsDirect, the free alerting service which sends tables of contents by L‘-ma“
`for Elsevier Science books and journals. You can register for ContentsDirect online at: www.eisevier-
`com/locate/contentsdirect
`
`U924-85?9(2D0109)18:3 1-m
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`
`(C0/r’eI1tS continued/i'0m outside back cover)
`
`shor communications
`Birth outcome following maternal use of fluoroquinolones
`H. Larsen, G.L. Nielsen, H. C. Sc/um/teyder, C. Olesen and H. T. Snrensen
`
`In vitro activity of linezolid against Gram—positive uropathogens of hospitalized patients with
`Complicated urinary tract infections
`H. Oncla, F.M.E. Wagen/6/zner, N. Le/In and KG. Nczber
`
`In vitro activity of linezolid against clinical Gram-positive bacterial isolates from Taiwan: an area
`with a high prevalence of antibiotic resistance
`C. T. Fang, S.—C. C/mng, Y.—C. Chen, S.-M/. Hsie/1 and W.—C. Hsie/1
`
`Letters
`
`In vi.::o activity of amprenavir against Pneumocystis carinii
`C. Atzori, G. Fanfoni. A. Valeria and A. Cargnel
`
`The cationic amphiphilic molecules as potential irnrnunostiniulants for HIV-1 infection
`A. Ac/tour
`
`Special Section: Antibiotic resistance: risks and strategies
`
`Obituary
`Torn Liergan (1939-2001)
`J.D W171/z'czms
`
`Original articles
`Antibi 31;lC usage in Nordic countries
`T. Bergcm
`
`The usage of antibiotics in Russia and some countries in Eastern Europe
`L. §":‘mtc/zounski, A. Bedenkov, W. Hr}/tniewicrz. V. Krcmery, E. Ludwig and V. Semenov
`
`The experience of reducing antibiotics used in animal production in the Nordic countries
`M. "Vierup
`
`Animals and antibiotics
`A.]l/1'. Johnston
`
`Antibiotic resistance in hospital pathogens~acquisitior1 or spread?
`JD. Williams
`
`HOW to organise antibiotic prescription
`P. Lellamonica, P.M. Roger, A.
`iM0usm'er, R. Collomb, E. Bernard and T. Fosse
`
`Erratum
`A. 1\‘0sz‘r0, G. Bisignano, M.A. Cannazelli, G. Cris, iM.P. German(‘) and V. Alonzo
`
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`
`This material may be protected by Copyright law (Title 17 U.S. Code)
`
`International Journal of Antimicrobial Agents l8 (2001) l73—l78
`
`Il\‘TEl'{\lATIONAL IOCl“\.\lAL (IF
`
`Anti microbial
`Agents
`
`vvvvw.ischemo.org
`
`Original article
`
`Comparison of in vitro antifungal activities of topical antimycotics
`launched in 1990s in Japan
`
`Kazuhiro Nimura, Yoshimi Niwano *, Seiji Ishiduka, Ryoichi F ukumoto
`
`P/z;.;‘I7'1a€€ufim/
`
`Research l)epm-‘Invent, Sam Plmrmaceufical C0,, Ltd., 8-5 Iligczs/Ii-()/Ii, 6-C/zome, S’/zilmgrlwa-ku, Tokyo 1400011, Japan
`
`Received 28 November 2000; accepted 6 February 2001
`
`In vitro anti-dermatophyte, anti—Candz'da albicans and anti-Ma/assezici fmjfi/r activities of amorolfine hydrochloride (AMF),
`terbinaiine hydrochloride (TBF), butenafine hydrochloride (BTF), ncticonazole hydrochloride (NCZ) and ketoconazole (KC/4). all
`of which were introduced for the treatment of dcrmatomycoses in the l990s in Japan, were compared. Although all of the test
`drugs are classified as an ergosterol biosynthesis inhibitor, the antifungal properties were found to be different. TBF and BTF
`exerted extremely potent antifungal activity against Tric/wp/1.yton spp. but not against C. albicans and M. fu7jf'ur, whilst KCZ and
`NCZ showed potent antifungal activity against C. albicans and M’. fmfur rather than Tric/mp/zyzon spp. AMF exhibited potent
`antifurzgal activity against all of the fungal species tested. Fungicidal activities of these antifungal agents against T. rubrum were
`determined by using neutral red staining. The fungicidal potentialities correlated with those obtained in the in vitro susceptibility
`test as determined by l\/IICs against dermatophytes. TBF, BTF and AMF exerted more potent fungicidal action than NCZ and
`KCZ.
`4'55 2001 Elsevier Science B.V. and International Society of Chemotherapy. All rights reserved.
`
`Keywnr.’s: Amorolfine; Terbinafine; Butenafine; Neticonazole; Ketoconazole: Dermatophytes; Canrlida albfcct/is; Malassezia fmfur
`
`1. Introduction
`
`A large number of antifungal agents have been intro-
`duced for the clinical topical treatment of der1natomy-
`C0568 [l—8], and in Japan several topical agents were
`additirnally launched in the 1990s.
`Included among
`them were imidazole, allylamine, benzylamine, and
`morplxoline classes of antifungal agents. Although all of
`[116 agents were reported to exhibit a broad antifungal
`Spectrum and to be highly active [9—l8], direct compar-
`ative studies on antifungal activities of these agents
`haVe rot been reported so far. In this report, we chose
`fiV€
`representative antifungal agents based on their
`Chemical structures and antifungal mechanisms, and
`
`1*Corresponding author. Tel: -81-36298-3017; fax: +81—3-3767-
`390.
`E~Imu/ acldrem: y.niwano@sato—seiyaku.co.jp (Y. I\Iiwano).
`
`compared their antifungal activities in vitro against
`Trichopliyron spp., Candida albicans and rllalczssezia
`furfur which are major pathogenic fungi of der1nato—
`phytosis, cutaneous candidiasis and pityriasis versi-
`color, respectively. The chosen antifungal agents were
`two imidazolc compounds (ketoconazole and neticona—
`zole hydrochloride), an allylamine compound (ter-
`binafinc hydrochloride),
`a benzylamine
`compound
`(butcnafinc hydrochloride) and a morpholine com-
`pound (amorolfine hydrochloride).
`Most of the antifungal agents developed in the 19905
`were also reported to be characterized by having fungi-
`cidal action [9—l3,l6]. Recently Fukuda et al. success-
`fully established a susceptibility test for determining
`fungicidal activity against dermatophytes by using neu-
`tral red staining which has been used for supravital
`staining [19,20]. We describe here the fungicidal activi-
`ties of the five antifungal agents against T.
`rubrum
`measured using the neutral red assay.
`
`0924-8579/01/S20 © 2001 Elsevier Science B.V. and International Society of Chemotherapy. All rights reserved.
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`l74
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`K. Nmmra el u/. /’Iliternmional Jourm/I ofAmimicroliial Agents 18 (3001) I73 —178
`
`ln this study, anti—C. albicans activity was measured
`according to the NCCLS M27—A protocol
`[21], and
`anti—iM. furfur activity by an agar dilution technique
`with Dixon agar that is commonly used for the culture
`of ll/Ialassezia spp. Anti—dermatophyte activity was
`measured by a micro-broth dilution technique using
`Sabouraud dextrose broth instead of RPMI l640 that is
`
`[22].
`recommended by the NCCLS M38—P protocol
`There have been several reports of in vitro susceptibili-
`ties of Triclmp/zyton spp. measured using Sabouraud
`dextrose broth and/or
`agar
`[l0,l2,l4,l6,l7]
`and
`Fukuda et al. reported the use of Sabouraud medium in
`the neutral red assay for assessing fungicidal activity
`against Tric/zap/zyton spp. [19,20].
`
`2. Materials and methods
`
`2.1. Organisms
`
`2.2. Test malaria/5'
`
`Neticonazole hydrochloride (NCZ), butenafine hy.
`drochloride (BTF) and terbinafine hydrochloride (TBF)
`were extracted and purified from the cominercially
`available 1% cream preparations, Atolant“"’ (SSP Co”
`Ltd., Tokyo, Japan), Mentax® (Kakcn Pharmaceutical‘
`Tokyo, Japan). and Lamisillfi‘ (Novartis Pharma K.K_‘
`Tokyo, Japan), respectively. Their chemical structureg
`were identified by nuclear magnetic resonance spectra;
`the
`purity
`of
`all
`test materials was
`> 99.0%:
`Amorolfine hydrochloride (AMF) was supplied by Ky.
`orin Pharmaceutical
`(Tokyo,
`Japan). Ketocorazole
`(KCZ) was purchased from Sigma (St. Louis, MO)
`The chemical structures of the test drugs are shown in
`Fig. l.
`
`2.3. In vizro sizscepzibility testing as cletermined bj
`A1]C5
`
`Twenty stock cultures of Tric/zp/1yin/2 spp.. five stock
`cultures of Culbicans and five
`stock cultures of
`
`iW.fm_'fur obtained From Teikyo University Institute of
`Medical Mycology (Tokyo, Japan), Institute for Fer-
`mentation Osaka (Osaka, Japan) or Chiba. University
`Research Center For Pathogenic Fungi and Microbial
`Toxicoses (Chiba, Japan) were used in this study.
`
`The minimum inhibitory concentrations (MlC,) for
`Tric/tophyton spp. were determined by a twofold micro-
`broth dilution technique using Sabouraud dextrose
`broth (SDB). Each drug dissolved in dimethyl sulhxide
`(DMSO) was diluted 1:49 with the same medium. A
`conidial suspension of each strain was prepared in
`sterile physiological
`saline
`containing 0.05% (v/v)
`
`Al‘/_F
`
`Fig. 1. Chemical structures of amorolfine hydrochloride (AMF), terbinafine hydrochloride (TBF). butenafine hydrochloride (BTF), netit
`hydrochloride (NCZ) and ketoeonazole (KCZ).
`
`rnaZ0lC
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`K. Nilnura et al. Intern£tti0m1/ Journal 0f/1nzirnicr0bi(t/ Agents 18 (2001) 173 178
`
`175
`
`TV\/E311 80 from cultures grown on 1/10 Sabouraud
`dcX[1‘OS€ agar slants at 27 C for l—4 weeks. Following
`filtration through a sterile cell strainer (pore size 40 um,
`BeC")I1 Dickinson Labware, Franklin Lakes, NJ) to
`remove hyphal fragments and agar blocks,
`the final
`coindial suspension was adjusted to 2 X 104 conidia/ml
`in SUB. One hundred mieroliters of a conidial suspen-
`sion and 100 pl of the drug—containing medium were
`added
`to each well of
`fiat—bottomed microculture
`plates. After incubation at 27 C for 7 days, the MIC
`was determined as the lowest drug concentration that
`pre\. .-nted visual fungal growth.
`MICs for stock cultures of C. albicans were measured
`
`twofold micro-broth dilution technique with
`a
`3y
`R131“ II 1640 medium buffered with 0.165 M mor-
`aholinopropanesulfonic acid (pH 7.0) according to the
`\lCL.‘LS M27—A protocol [21]. Each drug dissolved in
`Di\/I80 was diluted 1:49 with the same medium. One
`
`~
`
`iunclred ul of a yeast cell suspension (1 x 107’ cells/ml)
`and t00 ul of the drug—containing medium were added
`0 each well of flat—bottomed microculture plates. After
`incubation at 35 °C for 24-48 h. turbidity of each well
`was measured at 620 nm. The MIC was determined as
`
`he lowest drug concentration that showed 80% inhibi-
`ion of control fungal growth.
`MZCS against stock cultures of M. fiufur were mea-
`sured by a twofold agar dilution technique with Dixon
`agar (malt extract 4%, oxgall 2%, Tween 40 1%, glyc-
`erol ’).25%, agar 1.5%). The yeast cell suspension of M.
`furfur was prepared in Dixon broth from cultures
`grown in the same medium at 37 °C for 3 days, a.nd the
`final suspension was adjusted to 106 cells/ml. Approxi-
`mately 5 ul of each suspension was inoculated onto the
`agar plates containing a drug using a micro-plantar
`(MI"F"—P, Sakuma Seisakusho, Tokyo, Japan), and fun-
`gal growth was observed after incubation at 25 °C for
`5 days. The MIC was determined as the lowest drug
`concentration that prevented visual fungal growth.
`
`2.4. Ventral rec] cissay_f()r nteczsnringfungicidal activity
`in vitro
`
`To evaluate the fungicidal activity of antifungal
`agents against dermatophytes, neutral red (NR) assay
`‘ WES performed basically according to the method of
`Fukrda et al.
`[19,20]. A conidial suspension of T.
`Vubrwn TIMM 3866 was prepared in the same way as
`111 the in vitro susceptibility study to obtain the final
`C0nidial suspension of 104 conidia/ml. Five hundred ul
`Each of the conidial suspension was incubated at 27 °C
`FOT 4- days, and then the drug solution in DMSO was
`added to be a 1% (V/v). After further incubation at
`27 °e for 24, 72 and 168 h, NR (3-amino-7—dimethyla—
`H1011» =—2—methylphenazine hydrochloride; Merck, Darm-
`5/lfldt. Germany) dissolved in phosphate buffered saline
`‘PBS; was added to be a concentration of 50 ug/ml.
`
`A
`
`The cultures were subsequently incubated at 27 °C for
`1 h, and incorporated NR into the living fungal cells
`was measured. The cultures were centrifuged (14 000 X
`g, 5 min). and the precipitate obtained was fixed in
`formaldehyde/CaC12 solution (4% formaldehyde and
`1% CaCl3) following washing twice in PBS. Neutral red
`was then extracted in a mixture of acetic acid and
`
`ethanol (1% anhydrous acetic acid and 50% ethanol).
`After centrifugation (14000 X g, 2 min),
`the optical
`density of the supernatant was measured at 540 nm.
`
`3. Results
`
`3.1. In vitro .s'u.s‘(:eptz'bility of Trichophyton spp., C.
`albicans and M. furfur tr) antifungal agents
`
`In vitro susceptibility distributions of twenty strains
`of Tric/7.0phyt0n spp., five strains of C. albicans and five
`strains of M. ftnjin‘ to AMF, NCZ, KCZ, BTF and
`TBF are summarized in Table l. The lowest MIC
`
`against each strain of Trit'n()p/tyton spp. was obtained
`with terbinafine hydrochloride ranging from 0.008 to
`0.063 mg/l and the highest MIC with ketonazole, rang-
`ing from 4.0 to 16.0 mg/l. The antifungal activity as
`determined by MICs were in order of potency: TBF.
`BTF, AMF, NCZ and KCZ. These in vitro susceptibil-
`ity data of Trichopliyton spp. are similar to those re-
`ported previously [l0,l2,l4,l6,l7]. In contrast
`to the
`antifungal activities against Tric/20p/iyton spp., KCZ
`and AMF exhibited the lowest MlCs against each
`strain of C. albicans and M. furfur, respectively, while
`antifungal activities of BTF and TBF against these two
`fungal species were low.
`TBF and BTF showed potent antifungal activity
`against Trichop/iytan spp. but not against C. albicans
`and M. furfur, whilst KCZ and NCZ showed potent
`antifungal activity against C. albicans and 71/1. fmfnr
`rather than 'l‘ri(:/mp/zyton spp. AMF exhibited potent
`antifungal activity against all fungal species tested.
`
`3.2. In vitro fungzricial activity against T. rubrum using
`the neutral red assay
`
`In vitro fungicidal activities of AMF. NCZ, KCZ,
`BTF and TBF against T. rubrum were measured by the
`incorporation of neutral red (NR) into the living fungal
`cells (Fig. 2). In the control cultures, the incorporated
`NR (as indicated by the optical density at 540 nm)
`increased with time,
`indicating that living fungal ele-
`ments increased time-dependently. All of the test drugs
`reduced the NR—uptake in a concentration— and in a
`time—dependent manner, but the reduced patterns were
`different among the drug—treated cultures. TBF and
`BTF did not
`reduce the NR—uptake after 24 h of
`incubation, but thereafter the NR—uptake was progres-
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`K‘ Nimum er a]. /Imernati0na/ Journal 0fAr1limz'u'obz'c1I Agents 18 (2001) I73—]78
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`K. Nimura el 11/. /I/uerrmlional Journal of.4/1tz'nzz'rr0/7ial Agmls I8 (2001) 173- J78
`
`.1
`
`‘fable 1
`In vitro susceptibility distributions of 'l"ric/zap/iyron spp.. Candida alb/Tczms and .Ma/ase:i'afuifm' to amorolfme hydrochloride (AM1-'). terbinafine
`]]yfl1‘Oil1lOfl(l.C (TBF), butenafine hydrochloride (BTF), neticonazole hydrochloride (NCZ) and ketoconazole (KCZ)
`
`'1-he fl~;11]b€l' of strains inhibited at drug concentration (MIC: mg/'1)
`
`J Anti-I1lyC()1lC
`
`T. nielilczgrup/1yles' (l0)“ and T. rubrum (l0:)"‘
`
`0.004
`
`0.008
`
`0.016
`
`0.031
`
`0.063
`
`0.13
`
`0.25
`
`2
`
`14
`l
`
`4
`
`species
`antifungal activity against all of the fungal
`tested. It possesses wcll—balanced antifungal profiles.
`Fungicidal effect is also an important factor in evalu-
`ating the potency of antifungal agents. In this study, T.
`rubrum was chosen as a test fungal species because it is
`detected most frequently in patients with dermatophy-
`tosis in Japan. TBF had the most potent fungicidal
`effect although it required more than 24 h to induce
`fungal death. In regard to fungicidal action of TBF
`against dermatophytes, Ryder suggested that squalene
`accumulation subsequent to the inhibition of squalene
`epoxidation might play an important role in causing
`cell death [27]. Thus, one of the possibilities is that it
`takes time for squalene accumulation to cause fungal
`death. BTF and AMF also exhibited potent fungicidal
`action against the test fungus. As for BTF, fungicidal
`action is probably attributable to the squalene accumu-
`lation since the antifungal target is the same as that of
`TBF. In the case of AMP, Polalc et al. reported that the
`drug showed greatest fungicidal activity against T. men-
`mgrop/iytes. Of the organisms tested (T. menzagrm
`p/tyres, C.
`albicans. Histoplasma
`(:ap.s'ulatum and
`C'rypI0mc('us ne0f0rmcm.s') concentrations as
`low as
`0.001 mg/ll of the drug was enough to kill 90% or more
`of T. memagmp/iyzes when cultured on casitone agar
`for 48 h [9]. We used different culture conditions from
`those reported by Polak et al., but AMF also exhibited
`potent fungicidal action against T. rubrum. The fungici-
`
`0.063
`
`1
`
`t\mj—niycotic
`
`C. albicans (5)8
`0.016
`0.031
`
`4 M
`
`. furfur is)“
`0.003
`0.13
`2
`
`AMF
`TBF
`I BTF
`I NCZ
`KCZ
`
`Anti-mycotic
`
`AMP
`IBF
`
`I BTFNCZ
`I KCZ
`
`“' The number of strains tested.
`
`|
`
`sively decreased. NCZ and AMF inhibited NR—uptake
`at all measured points. Although NCZ strongly inhib-
`ited 1\I R-uptake at 24 h, at and after 72 h the degree of
`inhibition by AMF was more marked than that by
`l NCZ. The inhibitory pattern of NR-uptake by KCZ
`i was th 2 weakest.
`l
`
`4. Disc ussion
`
`As shown in Table 1, although all of the test drugs
`are
`ciassified
`as
`ergosterol
`biosynthesis
`inhibitor
`08,23 -26], these antifungal properties were proved to
`be different. What causes the differences in antifungal
`1 Pfoperies is not clear, but it may be related to their
`Chemical structures and/or target enzymes in the ergos-
`terol biosynthesis pathway. TBF and BTF, both of
`Which are members of allylamine and benzylamine
`Classes of antifungal compounds, and are known to be
`“lualene
`epoxidase-inhibitors
`and are
`very
`active
`aga111Sl Trichop/iytrm spp. but only have feeble action
`against C. albicans and M. furfur. In contrast, KCZ and
`NCZ, both imidazoles and known to be CYP5l
`(P4501.-—,M)—inhibitors,
`are potent
`antifungal
`agents
`against C. albiczms and 1M. fuijfizr. Of the antifungal
`agents
`tested, we noted in particular that AMP,
`21
`m]01‘pholine antifungal compound and an inhibitor of
`A4‘f€(1L1CIaS€
`and A8—A7-isomerase, exhibited potent
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`CFAD V. Anacor, |PR201 5-01 776 ANACOR EX. 2105 - 9/“IO
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`178
`
`K. Nimum er (1/. /Internaz‘iona/ Journal of Antimim‘0b1Ta/ Agents 18 (2001) 173—l78
`
`[10] HirataniT,AsagiY.MatsusakaA,UchidaK,Yamagu-chjH.In1
`
`vitro antifungal activity of amorolfine, a new II10l‘pl1fl.ll11€
`timycotic agent. Jpn J Antibiotics l991;44:993—l006.
`[11] Petranyi G. Meingassner JG, Micth H. Antifungal activity 0f[h\.,
`allylamine derivative terbinafinc in vitro. Antimicrob Agems
`Chemother 1987;31:1365 8.
`
`an
`
`[12] Hiratani T, Asagi Y, Yamaguehi H. Evaluation of in Vim
`antimycotic activity of terbinafine, a new allylamine age it. Jpn]
`Med Mycol
`l99l;32:323—32.
`13] McNeely VV, Spencer CM. Butenafine. Drugs l998;5.'.-105-13
`[14] Arika T, Yokoo M, Yamaguchi ll. Butenaline, a new benzi.
`laminc derivative: In vitro effect on arthrospores of T. }’7!‘€]Zrug)‘g],,
`p/tyres and therapeutic efficacy on experimental tinea pedis ir.
`guinea pigs. Jpn J Med Mycol 1992;33:541—7.
`[15] Odds FC, Milne LJ, Gentles JC, Ball EH. The activity in mm
`and in vivo of a new imidazole antifungal,
`l<ctoconazole.J
`Antimicrob Chemother l980:6:97—104.
`[16] Minagawa II, Kitaura K, Mineura K, Marumo H. Studies on
`antifungal activity of ketoconazole (KW—1414). I. In iftro anti.
`fungal activity. Jpn J Med Mycol
`l982;22:171—80.
`[17] Macbashi K, Hiratani T, Uchida K, Asagi Y, Yamagti-.;hi H.111
`vitro antifungal activity of SS7l7, a new imidazole antimycotic.
`Jpn J Med Mycol 1990;31:333—42.
`[18] Maebashi K, Hiratani T, Asagi Y, Yamaguchi H. Studies on the
`mechanism of antifungal action of a new imidazole atitiniycotic
`SS717. Jpn J Med Mycol 1990311343 54.
`
`i
`I
`I
`
`'
`
`l
`X
`
`[19] Fukuda T, Naka W, Tajima S, Nishikawa T. New antimycntic ‘I
`susceptibility test using neutral red to measure minimuzii fungici
`l
`dal concentration. Jpn J Med Mycol 1994;35:175—8.
`‘ed assay,
`[20] Fukuda T, Naka W, Tajima S, Nishikawa T. Neutral
`in minimum fungicidal concentrations of antifungal agentsl
`Med Vet Mycol l996;34:353—6.
`[21] National Committee for Clinical I.aboratory Standards. Refer-
`ence method for broth dilution antifungal susceptibilny testing
`of yeasts. Approved standard. NCCLS document M27—A
`Wayne, PA: National Committee for Clinical Laboratory Stan— '
`dards, 1997.
`.
`
`I I l
`
`National Committee for Clinical I_ab0ratory Standards. Refe1‘-
`
`ence method for broth dilution antifungal susceptibiliy testing
`of conidium-forming filamentous fungi. Proposed standard. NC-
`Cl.S document M38-P. Wayne, PA: National COIILs1tl1'::€
`for
`Clinical Laboratory Standards,
`l998.
`Polak-Wyss A, Lengsfeld H, Oestcrhclt G. Effect of o;...conazol:‘:
`and Ro 14-4767/002 on sterol pattern in Cana’z'du a/bicrmi
`Sabouraudia 19851232433 42.
`
`‘
`
`]
`
`Petranyi G. Ryder NS. Stiitz A. Allylamine derivatives: NEW
`class of synthetic antifungal agents inhibiting fungal
`sqrialenc
`epoxidase. Science l984;224:l239—4l.
`Hiratani T, Asagi Y, Yamaguchi H. Studies on antifungél
`mechanisms of action of butenafine hydrochloride II. ".‘on1Pa‘:"
`son in the response to drug treatment between a wild—type st1'éI1ll_l
`and tolciclate—resistant mutant strains of Sporor/zr1'.\'
`.t'/ztmckii.
`Jpn J Med Mycol 1991:32:139—49.
`A
`Van Den Bossche H, Willemsens G, Cools W. Cori
`lisscfl 1"
`‘
`Inhibition of ergosterol synthesis in Cczziciidzz cl/bier/rI.r by k9[‘’'
`conazole. Arch Int Physiol Biochem 19'/9;87:849—5l.
`1"
`Ryder NS. Mechanism of action and biochemical selectivity O
`allylaminc antimycotic agents. Ann NY Acad Sci 198555442205" ’
`20.
`Nishiyama Y, Asagi Y, Hiratani T, Yamaguchi H, Y:.\1113d3_‘\_l'
`’
`Osumi M. Morphological changes associated with grow‘ ‘ll lnlllb" .
`tion of Tl‘l'(’/Z()]9/2V1«‘l()71 menIagI'()p/tyres by arnorolfinc. Clin Ell‘
`]
`Dermatol (Suppl l) l992;l7:l3—7.
`
`dal mode of action of AMF has not been clarified, but
`
`direct membrane destruction might cause fungicidal
`effect, at least in part, as reported by Nishiyama et al.
`in an electron microscopic study of T.
`I’I7(,’I'lIagi'()p/Z_1?l€.S
`exposed to AMP [28]. Of the imidazole drugs, NCZ
`showed potent inhibition of NR—uptake but the inhibi-
`tion did not
`fall below the initial
`level
`(at 0 h of
`incubation) except the highest concentration (10 mg/1)-
`treated cultures, suggesting that antifungal action of
`NCZ against T. rubrum was possibly fungistatic rather
`than fungicidal. The inhibition of NR-uptake by KCZ
`was weakest among the test drugs as seen also in the
`MICs,
`indicating that
`the antifungal effect of KCZ
`against T. mbrum was poor.
`The antifungal agents tested in this study have differ-
`ent antifungal properties which seem to be related to
`their chemical structures and/or target enzymes in the
`ergosterol biosynthesis pathway. Of the test drugs, de-
`serving special mention is the broad spread of antifun—
`gal
`activity of AMF and the
`extremely potent
`antifungal activities of TBF and BTF against dermato-
`phytes. To further examine the causes of these differ-
`ences in antifungal properties, we intend to investigate
`drug-incorporation into fungal cells and/or interaction
`of stereochemical structure of each drug and corre-
`sponding target enzyme.
`
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