`
`BIOMEDICAL LIBRARY
`
`APR 0? 1997
`
`University of California
`Los Angeles
`
`ABSTRACTS
`
`of the 36th
`
`lnterscience Conference
`
`on Antimicrobial Agents
`
`and Chemotherapy
`
`an Annual Meeting
`of the American Society for Microbiology
`
`September 15—18, 1996
`
`Ernest N. Morial Convention Center
`
`New Orleans, Louisiana
`
`Amercan Society for Microbiology
`Washington, DC.
`
`Kaken Exhibit 2037
`
`Acrux V. Kaken
`
`IPR2017-00190
`
`Page 1 of 4
`
`

`

`©1996 American Society for Microbiology
`1325 Massachusetts Avenue, NW.
`
`Washington, DC 20005—4171
`
`All Rights Reserved
`Printed in the United States of America
`ISBN 1-55581-113-2
`
`Page 2 of 4
`
`

`

`CONCLUSIONS
`
`Abstract No. F792
`
`A new triazole, KP—103 had potent activity against 0. aibr'oans and
`moderate activity against den'natophytes. The anti—Candida activity of KP—103
`was higher than that of all the reference drugs. CTZ, NCZ. LGZ and BTF. On
`the other hand.
`the anti—Trich0phyton activity of KP-103 was equal to or
`higher than that of 012 and N02 but lower than that of LCZ and BTF.
`
`The anti—Tdchophyron activities of the reference dmgs were substantially
`reduced when cultures were grown in seam—supplemented medium or hair
`suspension, but the activity oi KP-103 was less affected. KP-los exhibited
`fungicidal activity comparable to LCZ and BTF against T. mentagrophytes
`when cultures were grown on the excised human horny layer. KP—103 has
`a low affinity with keratin as compared with LCZ and BTF. These biological
`characteristics of KP—103 might be reflected by its favorable in vivo etficacies.
`in summary, [KP—103 is active against a wide variety of pathogenic fungi
`including yeasts, dennatophytes. and Aspergr‘iius spp. Since it has a low
`affinity with the horny layer of the skin. its antitungal activity seems well kept
`in this tissue.
`
`For further information. please contact the following
`
`TAIHA OKAMOTO
`Mariagor
`Intemetlonel Operation and Licensing Department
`
`FAX :B1-4?3—90—8‘|-51
`
`KAKEN PHARMACEUTICAL 00., LTD.
`HINODE 1. URAYASU$HL CHIBR 279. JAPAN
`PHONE: 51 473-90-6140
`
`in vitro Activity of KP-103, a Novel Topical Antlfungal Triazole.
`Y. Tatsumt. M. Yokoo, T. Anita, H. Ogura. K. Nagai. and T. Naito.
`Development Research laboratories, Kaken Pharmaceutical 00., Ltd, Kyoto, Japan.
`H. Yamaguchi, Tetkyo Univ.. Tokyo, Japan.
`
`ABSTRACT
`
`The in w‘tro activity of KP—103. a triazole having 4-methylenepiperidine moiety at the 0—3 position.
`was compared with that of ciotrimazole {0T2}. noticonazole (N02). lanoconazole [LCZL and butenatine
`{BTF} against pathogenic fungi. Nile values (ugimll were shown below.
`
`M'Coo semi)
`.
`BTF
`LCZ
`NCZ
`CTZ
`(No. of strains) @403
`Fungi
`:80
`0.25
`0.0825
`0.0313
`C. albicans (44)
`0.002
`12.5
`0.78
`3.13
`6.25
`M.
`fan‘er (6}
`0.025
`o.25
`0.002
`0.25
`as
`Aspergiilus spp.[15)
`0.0625
`0.0078
`0.0078
`0.125
`0.5
`T. rubrum {39]
`0.125
`0.0156
`0.0313
`0.25
`o.2s
`T. mantagrophyies (as) o.25
`a A. 0.165 M MOPS-buffered HPMI 1640 medium. pH 10; B. Sabouraud dextrose broth:
`C. medium C (Faergemann. J. at a]. Acta Derm. Venereol. Suppl 86: 143, 1979).
`
`. a
`Media
`A
`C
`A
`8
`s
`
`KP~106 was the most active against C. albicans and M. fudur among the tested drugs. Its activity
`against Trichophyron spp. was almost equal to those of 012 and N02. but was weaker than those
`01 L02 and BTF.
`Anti—T. mentagrophytes activities oi the reference drugs were reduced by the addition of human
`serum and homy materials as reported. while that ol KP—103 was not affected. Furthermore. anti-T.
`meniagropytes activity at KP—103 on the stepped human homy layer was equal to those of L02 and
`BTF. These results reflected in vivo efiicacies.
`In summary. KP—103 has a broad antitungel spectmm and could keep a high activity in the homy
`hyer where iungi reside.
`
`Figure 1. Chemical structure of KP—103
`
`
`
`36th lntersciehce Conference on Antimicrobial Agents and Chemotherapy
`
`

`

`Table 7. The role 0! adsorption lit KP—ttta to keratin
`
` me 1 herein ratio MM M—
`in the incubation mixture
`109—106
`LC:
`BTF
`1:5ooo
`50.:
`91.9
`95.1
`
`Figure 3. The rate of rile-n ot' KF-103 Iran the dug-hound heath
`—-
`KP—toa
`i
`u to:
`31
`I33 ETF
`
`U E E
`
`.3
`H=
`E=
`u
`
`
`
`Tln'leeciweeh
`
`RESULTS
`
`1} Against Conical-1.9.012.ch LCZ. end BTF were 15—. 82—. 125—.
`54096—1010. respectively.
`less active than rte-103 friable it.
`flAgahist Khmer. KP—lmwasiB-foldleesmfllan chandBTF
`but as active as NC: and twofold more ecuve than C12.
`Mamet T. menlegrcphm 1CP—108 was 0— to 16—fold less active than
`LCZandeTthrasactiveasc‘lZanleiizrrableti.
`
`3} Tile 11thso value oi KP~103 was equal to or two times mortar than its
`MIC” value tor TWW spa. which indicates
`that KP—103 was
`hingiclrlei against this fungl (Tables 1 and 2t.
`41' Wt Candide. Winsome. and Helm 5w. KP—ti‘la was the
`most active among the tested drugs treble 3].
`nmmmem.KP—1oameeemnemenmzmrmore
`ecthred'mmlilcz. and BTF linear.
`61 MW! eenneiophyies ctherthen in whom and ‘1: MW
`KP—ioswaslessactiyetmzandBTFhulmoreectivetnanmmd
`ncz {reins 5:.
`Ti The anti—T. momma activities of the reference drugs were 3—
`lio ZT—lokt
`reduced by the addition 10% human serum to the assay
`rriedltl'l'l. but the activity of RIP—103 was little effected {Twin 41.
`at The anti-T. menlagrophyres edivitles ol' the ralarence drugs in See
`were 15— to 524010 more lower than ti'roen oi the reference drugs in are
`human hair srrapenaion in saline. out the activity at KP-toe was tile same
`In both media [relate 5}.
`'1'.
`5. m and
`against
`ersivtty
`9} KP—103
`had
`fungicidal
`riteniiegrephylesmwnonnumen horny layer.Armndthe drugs tested.
`KP—tntr was
`the met
`active
`agaii'ist c antenna. Against
`T.
`mentagmphyles.meestiytlyolKP-toe was stronger titan malolC'lZend
`Wmdmmwmmoiwzmmflablefii.
`10) KP—ttts showed low adsorption to keratin and high release from
`keratin e5 colt-lowed with Luz and B'I'F [table If and Flame 5}.
`
`REFERENCES
`
`Tattle 3. Antltungel activity of Eli-10:3 agallm vellum oatnonanic
`lunul
`
`organisms
`Geometric mean MIC gflg
`KP-103
`BTF
`(No. or strains!
`crz
`N02
`LC:
`0.0151
`$1064
`c, Wits is]
`0.1249
`02973
`03536
`0.11m
`1.4142
`c; horse! :2}
`0.125
`025
`1.4102
`0.0197
`>2.5iee
`c. perspeiiosls [at
`0.125
`1.2599
`1.5074
`0.0030
`4.0
`G. MW 11]
`0.00%
`05
`025
`elm:
`0.5
`c. slalom [ii
`0.125
`0.25
`0.120
`0.0313
`0.25
`6. nine ill
`0.125
`0.25
`00525
`
`“13‘
`C m“ ‘5’
`°-m3_°_"l‘_53__3£‘_93_.i“£..-
`Trimmer-ice 14) Wfi'fimufifie
`0.3535
`0.25
`0.5
`iflm'“£935.........‘LL-...‘;‘1_____E§____:‘EE___
`in. W re]
`0.025
`3.9415
`1-1015
`00191
`0.921:
`M MW’JNEE....‘.;fi-__°iL__El_._._E19§J-
`
`'rT'videos---'351-12:
`0.0155
`0.0004
`00221
`0.0014
`0.0059
`r. alallol [1:
`0.0313
`0.125
`0.0525
`0.0075
`0.00m
`in. carols in
`0.0315
`0.25
`00525
`00an
`0.0075
`111'. mourn :21
`0.0422
`0.1700
`00825
`0.0028
`0.001%
`E. 00W [1]
`0.0078
`0.0525
`0,0155
`0.001
`0.0078
`
`A. Wins is}
`0.0405
`1.2599
`0.1904
`0.
`10
`0.1904
`0.0011
`A. nevus l5}
`010413
`0.0590
`0.25
`nodes
`A. alga! :41
`0.0925
`an
`0.8586
`00024
`one!
`A. serious :21
`0.0m
`20
`0.3535
`00020
`0.1760
`A. mom :1!
`0.125
`2.0
`025
`0.001
`0.5
`
`Table 1. EM od' hum senate on anti-'1'. Wyatt!” eclltrtty cl
`KP-‘iM
`
`Geometric mean MIC Emu
`one
`see will 10% serum
`0.1487
`0.182101}
`c.1051
`amount
`
`1%???001 WW“;
`ire-sod
`012
`
`0.5535de
`0.13
`{film 1202
`“realm:
`noose
`LCZ
`
`on: nominal come
`
`
`Table 5. Efleot 01 lumen ltllr on anti-T: mental:qu activity 01'
`KP—tt‘la
`
` see
`591. hei‘ mason
`MIC
`MFc
`n'Il
`MIG
`MFG
`“momma
`02
`0.2
`0.2 on}
`poem}
`KP-103
`0.2
`0.39
`525032:
`12.5 0132)
`(:12
`0.1
`0.1
`moms:
`3.130321
`its:
`soon
`0025
`n1 1:15}
`cosine:
`L02
`
`BTF 1159054: 0.005 once 0.2 0321
`
`
`
`
`llbilcar'lil and '1'.
`Table 0. Furtglelddl activity 01 KP—103 Inelnal G.
`mummies: grown on humn llnnliir layer.
`_
`mm m
`Omenfims
`Human tea—m! 1521 M
`No.3
`No.2
`No.4
`none
`none
`0.1100
`31.0
`0.5
`)1.0
`
`Corripouride
`KF-ttlll
`C12
`
`No.1
`0.000
`>10
`
`No5
`0.003
`0.5
`
`gaff?“
`
`Ex“
`
`Lee
`N02
`BTF
`KP-103
`012
`N82
`Lt'ZZ
`
`51.0
`51.0
`>1.o
`0.010
`mi
`0.25
`0.016
`
`>1.c
`51.0
`51.0
`time
`51.0
`0.2.5
`0.015
`
`31.0
`51.0
`51.0
`0.010
`51.0
`
`51.0
`>1.o
`51.0
`001:
`>10
`
`51.0
`>10
`>1.0
`0.009
`05
`
`0.015
`0.125
`
`0.032
`05
`
`0.008
`0125
`
`1} Yalnagtlchl. H.. K. Ucntda. H. Kettle. T. Shi'lnrte. K. wetenaoe. T.
`Kueunolri. M, Himme. and H.
`ishlzaiti. 1995. Fieport at committee or
`clinical laboratory standards—1994. Jpn.
`.J. Med. Mycot decal-es.
`2} Faergernarin.
`.J. and 5. Ben-lender.
`1979. Tinee verelcoior and
`Pintoeporum more: rrtyeoInglcal Investigations. departmental infection
`and epidemiological smeye, .Acte Det’nt. Vermont Suppl 06: 1—23.
`
`INTRODUCTION
`Flecently. several kinds ct topical artilungal migs. such as Imidazolee
`{bllcnazole neticonazole. ketoconexcla, and Ianoconezoiei. allylemine
`{tertn'nelinel benzylamlna lbutanafinel. and mcrpnoline {amolfinel
`have been developed and li'illoduced 0110.019 market These are
`the once—s—day
`treatment
`at derrnetomycoata
`Altrmgh all at these antihlngal drugs have relatively broad spectra 01
`activity. tray very In the ardent 01 their activitles against diflerent fungi.
`larbtnellne and mistine am lees adtve against yeast—
`lungi. hut more active against derrneiophyles
`than are most
`There are only two tnazo-ie anlllungat agents.
`fluconazole and
`raconezote. which are used systemically tor the treatment 01 both
`deep mycosis and wei'ficlai mycosls. but not wed tcpbalty.
`rte-103 is a meet topics! lliazole having a rried'iylenaoiperlzine at
`In this my. we 813015!“ the in vino
`antil‘ungal arslvlty e‘l KP-Toe in comparison with that of the tolerance
`drugs [Figure 2:. clotltn-lazole lCTZt. netlccnezoie lNCZJ. tanoconszcle
`
`MATERIALS AND METHODS
`Susceptibility testing procedures.
`Harmonia and Wm spp.
`lilies were determined by the acme-based mlcrodilnt'lon rrietllod “.
`The test. medium used “'85 RPM 1H0 medlLlI'l adjusted to pH ‘l’.0 with
`0.155 M MOPS. The test unanlame were grown on yum morphology
`agar plates at 35'!) for 2 or 3 days. The colonies were amended |.I'I
`hlrcldlty 01' a 0.5 McFarland standard. The
`suspensions were diluted too-rain ror Candida app. and ill-rain for
`01pr app. with FIPMI 1540 medltl'n. Aliquots or 111 ml at the
`suspension were dispensed into the wells containing 0.: mi of two—told
`lnoculun'i
`size: Correlate 5pc:
`its—2.51110a
`cetisrrnl. CW 300.: Dis-2.5:"? cellar‘ml and all nila'oplalee were
`unedited a 5’6 for 1—2 days [Canada app.) and 3 days {Crmlococcm
`
`MICS were determined by the mlct‘odllutlon method using HPMI 15-10
`medium. The test organisms were grown on potato dextrose agar {FDA}
`slams at we for 10 days. Final innsutum slze was ixtn‘ coniolahtt All
`mlcroplates were trimmed at 30‘s for 3 days
`MICs were determined by the microdlmtion method using Saeouraud
`‘Ine hast organisms were grown on slants or
`glucose trloticese yeast—amact agar and Sabcuraud dextrose agar £50.91]
`at ao'c. Fri-lei Inocurum size was 1x10‘ conlcllan'nl or 1x10“ hyphanrmr. nil
`micrcpietes were incubated at 30'C for 7 days.
`Mitts were determined by the agar dilution method. Test organisms
`were grown on slants 01 FDA containing 1'95 yeast-extract. 1% peplnne.
`and 2% olive oil at ao'c tor 5 days. Five rnlcl'oltlela [1:10‘ cells: or the
`fungal suspension were spotted on rrledlnrn C 2' plates comlntno two—told
`serial dilutions and all plates were incubated at 30'C for 5 clays.
`Endpoint of miniature Inhibnwy and fungicidal concentrations [llths
`Mice tor Candida and ciypiocaccus mp. were defined as the lowest
`drug concentration lrlttihlirig :00 “in 01' growth competed with tile gram
`controls. title tor Molassezia sop, dermatopnytas. and AW app.
`
`:erf: dire-led as the lowest drug concentration that lnhltiited visible growth
`MFGs tor derrnaliophytoe were detennlried by subculture tit 10 Iill ~01
`broth on SDAplata at M‘Cbri'dawandweredellnedastnalowest
`drug concentration that produced 3 90% reduction at the fatal Inoculurn
`511001. of serum and hair on anti-T. :1“!qu activity.
`M65 of T. mnngmpl‘iytea were detenntnert by the mlu'udtlutlon
`three tested media 1503. SDB containing 10% human
`serum, and 5% human hairsuepeneion In saline). MFCs were determined
`by sthculture oi hair on son plate at 30'C tor 7 days and defined as the
`lowest drug concentration that completely prevented viable growth at fungi.
`Fungicidel activity on horny layer.
`Horny layer 01 the skin were stripped lrorn arms 01 M healmy male
`volunteers with 08110000110 tapes. The horny twat tapes were out. put on
`slide glass. and hurtlalerl at SO'C for 24 h. Twenty nticrdilter 01 two—told
`rlnld dilutions was applied on each horny layer and spread.
`Uneheorheo drug was wipe oil. alter each aloe gees was inctoated at
`room temperature 101-1 11. 5.01 15310:I cells] ol’llie lung-a1 suspension was
`spotted on each horny layer. Each slide glass was imitated at 50's for
`2 days. htch were determined by emporium of tape on 30.0 pets at
`'1? days and defined a the lowest drug concentration that
`
`anti-lily with hermit.
`Niouot at 0.1 oil of each drug aotuticn {i mgirnli was dispensed into
`9.9 ml at 5‘16 keratin swpanslon in saline to give a final concentration at
`10 mlml. Each tube was indicated at 31"!) for l h with shelling. Alter
`Incubation. me mtrtture was centriiugad and one isenl pardons ol' the
`supernatant were taken to determine the adsorption rate of drug to keratin.
`The Wily—bound herein was washed 10 lines by shelling in saline at
`3?‘C liar 10 min. Alter each wit. the mixture was centrifuged and ter
`150 pt portions or the eirpemetenl were taken to determine the release
`rate 01 ring tram keratin. The drug conceiwallrln In the amematent was
`determined by the conventional agar-wait mien assay titling A.
`ringer
`lorch. timeneagropnylieshre'rF. and alien-riorW—ioeesthe last
`eroefllern.
`norm 2. Chemical atmctllm til Mme: dnma
`
`-.
`
`Q "3
`
`Clonimezole {cm
`OH
`H
`
`3'5 I
`
`NAN\=l‘
`OCan-HCI
`Nellcenazole [1162}
`
`NC
`
`5
`
`“SNHSJ/CE
`
`Lenoconaziole lLCz‘l
`
`l
`N
`
`'HC‘
`Butermltne {ETFt
`
`Tahoe 1. Amil'ultael eclhrtty of KP—103 against C. allitch '1'. militant.
`and T. mentogmpl'lytu
`
`'
`Mic
`
`Han e
`.
`one
`dimers}
`mpmm
`
`00005—00150
`0.002
`0.002
`C. means
`KP—103
`[44}
`C12
`00078-025
`0.0155
`0.0813
`ncz
`coals—51.0
`0.0025
`0.0525
`LUZ
`00313-310
`0.125
`0.25
`BTF
`:00
`>80
`3-0.0
`
`T. Morten
`KP-100
`03158-105
`0.0825
`0.125
`1393
`0‘12
`00625—10
`0.25
`0.5
`NCZ
`0.0155—0.5
`0.0025
`0.125
`L102
`00005-00013
`0.0030
`0.00?!
`BTF
`00089-00156
`0.0030
`0.0073
`KP—ioa
`dudes-0.5
`025
`025
`on
`01125 4.25
`0.25
`1125
`NCZ
`0.0313—025
`0.125
`025
`L02
`0.001 41.0525
`0.0155
`0.0313
`
`00030-03150 0.0070BTF 0.0155
`
`
`
`T. molagmplams
`1251
`
`12 mentor and 1'.
`Table 2. FunaloldeI activity at KP—tt‘le against
`WWII
`'
`MIC
`1.
`$3053....
`I min-rim
`139}
`
`A
`a e
`swim“
`"-25
`0.125
`Maw-1.0
`KP-1W
`“.5
`0.25
`03625-20
`01?
`0.25
`0.25
`013156—11]
`NC:
`LCZ
`“DOCS-0.053
`0-00”
`0.0313
`BTF
`ROUGE-0.0153
`0.00m
`0H)?“
`
`1'. r'r'leni'egiropi‘lyt‘eia KP-103
`0.125 -1.0
`025
`025
`123:
`C12
`0.125 —0.5
`0.25
`0.5
`"CZ
`0.0525110
`0.25
`0.25
`LG:
`utilise-0.0625
`0.0313
`0.0313
`
`