Fosfomycin does not reduce cytostatic activity of cis

Short Camntun;catian
FOSFOMYCIN DOES NOT REDUCE CYTOSTATIC ACTIVITY
OF CIS-PLATINUM ACAINST HUMAN OSTEOSARCOMA
CETL LINES IN VITRO
Christoph Bührer.
MD .
crnil.Lren,s Hospnal. Irei€ univeßität tscdin. D \'\, 101]0
Jens Atzpodien, MD : Deparman of H€narolog,v an.r oncolo$, \r.rririnisthe
H..lrschule Hannover. D W 3000 Hanno!.f 61. Ce.manv
SadikOz,MD ! D.faf,D.nr .1I
crnal \4edicile
IlI,
Unjrersitätsklinik li.lrnsar.
D.ur 8520 Erlanser, Ge.Fin,v
GÜnter Henze,
.
MD . aihil,l...'! Hospital.
||€ie Ufiv..sirlit Be.lin, D \\'
lfol
Be.lin
\t\tn h t,tinnLt llt iat ntuLtn! at4fu,r4 .nn nlhnLrinj t\i&tn
.! ?tahnü Cn ..t ß rt la:ja\ttn i h !tuLrmtn t trt t@Ljl
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tu;üa idt 4l4t! 1nnttuttJw\tuxt d4tirütt ti.J !j7üß-tituit Lititit) mtlnn{üatin; ür
ünanitt. tt thtt L.jtaidat drt it) ii Li4iati"ln asa'd htnnn ßta:diüna .tr ht\ i ; ut atritttd b)
ünaß .a"untt.1iani aJ-lar;nt)ün 4 h a I n!,,nt in tiha Thi' t'sssß tA in tt^t /11 iltr iJ na
lnt' th.ntuttuttJnann nn-Lhn il n! tnrLtntr dl l.)!lant,1n tnh tgt.t ta nühdniiü ürti.l
Fr\furnran tat btn
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INTRODUCTION
In 1965. Rosenberg er al r€ported that bacrerial .:cll division can be
inhibncd bv electrclvsis products liom a plariDum elecrrode.' Subsequendv devdoped platinurn compouncls ba\.c bccn dernonstrarcd to possess clinical effectryeness asainst rarious cafcinomas and sarcomas, arrd cis-di.hlorodianminc
platinum-II (DDR cis platinum) is nou' bi:ing .,onsidered as first line
r.ll
!\t graren,]l] a.kn.r'l.dse T Piesdr. Hxnnovrr. Ccrnrfr l. r Fr.filiDg ih. h!J.rn.ne.sr.rna
line SK.OS 10 atrd B Sdrolz. Hamburg Grrnar! fur prori.liig rh. hln,rn.n..sar..n,3..ll liie
TI
35
udrq.x llntabhq ard anih-1\
3
2!3 211
1991
Cqyilltta l99l t| lltnqtüt lrblt:h|i! Ctlrntnr
243
C, BUHRER
ET AL
chemotherap,v in tcsticular and ovarian cancet adlanced squanous carcinoma
ol head and neck, and ostcosarcoma in children and adolescents.
Of its various side effecn, *'hjch incLude nausea, rorniting, myelosupprer
sion, and peripheral neLrropath)! nephroto:icitv and ototoxicity appear to be
thc most lnniting in administering cis-platinum. Renal Dnpairmcnt alicr cis
platinum is widcly asuned to be revefsible and r:an br: miriqared to some
exrent by hldration. mannnol induced diuresis, and cis piatinun admjnista'
rion in h,vpertonic saljne.'' Ho*'evcr, the clfecL of thesc measLrres on the rherapeutic €llicac\. ol cis platirrum has not becn dctermined Cis platiDum kills
ourcr hair cclls (prcdominantlv o{ rhe basal mrn) of the Corri organ in rodcnts'
ancl human patienrs,'rvhich rcsults in permanent hearing loss mainlv jn the
ligh an.t ultra'high frequ.".r' .""g..' " " O"" nfthe authors wirnessed a case of
rapid onsct dcafncss in a l,vear-old girl wirh ostcosafcoma alier a single i L,sion of cis platinurn.'!
Foslbrn,vcin (1,2 epox,vpropvl phosphonic acid), a broad spcctrum antibi
otic thar acts b,v inrerfering vnh bacrerial ccll rvall svnthesjs, has bccn shown Lo
ameliorare cis platinurn-medjatcd kidney and inner-ear darnage both in experi
mentai anjmals. "''' and huma" patienLs.' fhc data Presented here doron
strare that lbsfomyci. orcr a broad rangc ol concenrrations d.res not interlere
$'ith the abilirv of cis-plarinum to l l activcll divnlnrg human ostcosarcona
MATERIAL AND METHODS
Call
alrar. Thc hunan
osteosarcorna cell lmcs, SK-OS 10 and TE-85 $efe
RP\lI
l6a0 (Gibrc BRL, Eggenstein, Gerrnarrv), \upplenente.l with 2
mNt l--gluramine, 100 pgl'nl streptom\cin, 100 U,'nl porn:lllin, 25 rn\{
Hcpes, and 10?i Iital callserurn (PAN, Aidcnbach, Germany). ,\ll incubalions
kepr in
rverccarricdourina{irllyhunri.tiüedatllosphcrcol15%air/5%COraL37'Cl
Iosfom,vcin (Boehringer \Iannhcim, Nlannheim. Gerrlanv) and cis plaLinum
(N{cc]ac. Harnbu|g. Gernany) (,rrc diluted just prn,r to usc as indicate.t.
iH tb?nid;n innlpatutian
lnlt lation as\at. Cells culturcd for 72 hours in the
prcscncc or abscnce ol loslbmvcin and/r,r r:is platinum ri'ere pulsed for an addiriona] 16 hours with 'H{hymi.line (5 []i/mnrn, ,A.m€rshan Buchler Braur
schweig, Gern,aD\') \\h.,le cells \!efe harvesrc.l onto glass fibcr stfips. waslicd
irith x.atcr, and incorpor,lted ra.tioacrivil,v i\as measurcd in a p-counter after
.lli i'' |"i
.i'' ri,r". .,.o'.lt e ior.l
Cahiwtric riability ai:a1 Wetabolisrr of thc vellow dvc \ITT (3 (a.5
dimethvlthiazol-2-vl)2,5'diphcn,r'lrctrazoliumbfomid.l b) livc nitocliondria,
rrhich resulred in a color change roward violet, rvas uscd to cstimate the nunber
of riabie cells as desüibed.'' After an 84'hour cufture pon,d, cells wefe pulsed
rrnh ycllorv \4TT (Sigrra. Dr:iscnhofen, Gcrman\') at a final mnccDtration ol
FOSFOMYCIN DOIs
NOI
IMPAIR OSTEOSARCOMA KILL
245
0.5 mg/nl Ior.t hours Cells r"'ere spun .losn. üe sup,rrnatllnts removcd. and
vnner cNsrals of rcduced NITT werc disvnveci nr 100 pl isoplopanol containing
rcarl in an ELISA reader L,sing \\,ells
0.04 \ HCl. -A.bv,rbance at 595 nm
'!as
widrout cells as blank.
RESULTS
Viabilitl and proLiferarion of the human osrcosafcoma cell lines, SK-OS 10
and -lE-85, appearecl ro be sensitiic to the influen( of cis pl:rtinum. At r:is
platinum corcenLrarions ol or abovc 5 pgtrLl, no viable cells could be detecred
mrro*r,pn:alv alier 72 hours. and \l l T ri,duction as in.tistingLiishable lionr
background levels (Fig. 1). \'\'hcrcas no rmpacr on \4TT reduction was seerl at
cis platurun concenlfations belorv 0.6 pg.tml.'H th-,-micLinc incorporation was
signilicantlv reduced bv cis platirlum conci,ntrations as lori' as 0 15 ptlml (Iig
2). Therc was no rnajor dilftn:nr with .cgar.l ro cisllarinum scnsitivLt,v bet\leen the t\!o.ell lines us€d ahhougl the rncaD cis-platjnum .r)ncertrarrons
ncccssarl to achiele a i0 -'Z reduction ol Nll l or'Fl-th,vrnidine inmrpor:rtion,
respcctivclr, appeared io bc some\\,hat Lowef in rh.r a.lherenti,v grol'ing <rtl lne
SK-OS 10 than ur nonadherent TE-85 cells (clata nor shown) Iioslömlcir rLP to
I mg/rnl r.as wcll
b,v SK OS 10 and T[ 85 celLs Additnli o{ löslornv'oleraled
cin al final con.cnträtiorrs ben',een 20 :r d 500 pg,hl did not change thc dosc
response cunt n) cis plärinum \\'ilh rc{ard to \lTT reduction or incorporation
of radiolabclrl thvmictine (Fiss. I and 2).
DISCUSSION
The linlning orotoxicity and ncphfotoxicilv of thc antintlplasrir: agcnt cis
platinum nccessitares the investjgati.rn of means to recluce these toxicitics rvirh
our dc.rc^sing rhc tumoricl.tal polential of cis PlarinLrm. Fostom,v.in is a highlv
polar lor-rnolecular+veighr (r)mpound (138 D) kno"n to act specificallv
against thc bacterial enz)me \-aciitvl gtucosaminr: 3 0 enolpvruvvl rransferase
s)nthesis. \\rhen given
that is involvcd in the i ial phase of bacterial cell
'!all has
bccn shown to be
to rodcnts ai dosages of 300 Io :J20 ni:l/kg, losfom,vr:in
parriall) protecti\e againsl cis-plarinum mcdiated inner ear and kidney dam.g"." '' " Iu.f"-y.i" gi'en at dosages of 2 s tMicc a day to adult canccr pa
rients has been demonstrated rc reduce cis-plalinurn-relarecL nephrotoricit\:' '
platinum and lbslbm,vcin has been
However, clinical coaclminjsration .Jf
',is
c,lrrie{l our dcspite the patLcitv of experimoltal investigations evaluating the
antitlLmof efficac,v of cis'platinum in combinatjon tith loslbmvcir. In N\'IRI
mice, fosforn,vcin (500 mg/kg gnen intEpcritoneall,v) has been rePoned to increas€ rhc dose of cis-plarinum nccessar,v to cure 507o of anima]s injecrecl wih
svngeneic S 180 ascnes sarcoma r:cLls lrom 2 0 mq/kg ro 3.: nrg/kg.'' The same
246
C. BUHRER ET AL
1r
20 10 5 2a' l? 06 0! 012
Cis I'laiinum conccnt.r alron (ps./Inl)
f*r"-r- 50. ra/-,
I {,r,*' r""r"-y.r"
=
l:ll f..r""rt, ,oo Is,,-L
I
r,osroDy.- 10,ia,,L
006
FIGUIIE 1. \lTT ren!.ii.d,r'l u,1r r5rc!s&lnrr ürll lurt Tl8r ftalrd \nh rari.!s.Irtotrnronr.i
.ßpr',ounmdlor.nr.i.i.,rr.'al.rllh!r,\Erin,.rr\,.6rtxdnrrIIIS\r:t.r\,rhii!5.n]
Crs I:'lal.inun, con( enLr.it ion (t)g/rnl)
I {rbour r.sromvrn
l l r..r"-,"i" roo 1,s,,-r
=
E
F..r,--i.5oo pc/-l
n",."-"". :o |q/-L
IICU&E 2. 1l rhr di.e in.oDoriLiln olnulrxn lreosmom! rel lnt sK.os r0 L'.ätrd \nh v:i.!s
...(.iLrrtrns ot.is Fladnrn rnd a.\iro!rin 1o. ä rotal !133 houL
FOSFOMYCIN DOES NOT IMPAIR OSTEOSARCOMA
KILL
247
authors lbund thar simuhaneous inrraperirorleal aclministration of lbslomtcin
(500 ll)g/kg) aDd cis platinum (0.5 to I mgikg) in Wistar rats injccti,d wnh
\i,shida asr:ires sarcoma cells reduc€d rhc incrcase o{ lile span aftcr cis platinum
bv more than 50ti whereas fosfomrcin had hardll any inlluence on trearnenr
resuhs ol cis-piarirrlLm against L 1210 leukemia in DBA-2 mice. The dosagc of
cis-platirlum used in these rodenr cxperinerlts (500 mg/kg) was about 10 rimes
higher than that u$rd in r:linir:al tri:rls but conparable wllen cälculatcd accordinq to bodl surfacc arca. Fosfomvcin rvas injected dirccdv into thc tunorbcaring perironeal cavit\,. and local .oncenrfations arc likelv ro haye exceeded 1
ng/nn.
-fhe
experiments prescntcd hcre show thar in \itro tbslom\cin at conccntra'
tions up to 0.1 ng/ml does nor inrerfcrc idth thc capacitv ofcis plarinum ro kill
dividing human osteosarcona ce1ls. Tr can be assumect that thc ranec of fosfomvcin concenrrations cmplo,vcd in our in tiro experimenrs colers rhose
a.:hie\cd in clirlical sirualions: 15 minutcs alter inrravenous infusion of 2 grams
of fosfom,vr:in, scrum leiels have been shorvn to peak at nearlv 90 pg/nl and
then to ctecline rapidlv rlth a selum half-lifc of 1.5 ro 2 hours.'
The diflerence h spccics o gur and ltpe ofmalignanc\ might conrribLrte Lo
thc.liscrepanc\ ber\\€€n the results of\\:agner eL a) ancl those pr,:sented bcrc. Ir
caD be speclrlated rhär prorection against cis pl,ltinL,m-meclialed jnjurl- bv lbsfonvcin rcquircs üe availabilitv of certain cnzlnes or co läctors and is higNy
tissue specific. Scveral lines ol evi.lence suggcsr rhat nephropr.rtection and oto
prorection bv foslömrcu is a result of pharnacod-vnamic ratlii:r than pharmo'
r:okinoi< intcrar:rionr
1. ,\lthough fosfom|cin N nor mctabolized and js almos' corrlplerel,v rccov
cred in thc urinr: 2+ hours aft€r ntravcnous intusion iD humans,! cffectne
nep)rroprotection in rats has been achieled bv fosfomvcin preteatmem up to 24
hours prior to cis plarinun adminisrraLior
2. Fosfomvcjn has been shorrn m reduce damage to kidnev lubule and
inner-ear hajr cells by a variet,v of drugs besides cis'plarinum, such as
am;ro"-l).oside antibiorics,'' c,vclosporni,A..'' vancom,vcin," :rnd other glyco
peptide anljbjoLics.'
3. In experiments on animals. lbslomvcin has been demonsrfated to ftcely
cliffuse flom the nictdle car cayity to rhe perilvmph.'' Whcn fosfom,vcin is direcrl,! applied to the miclcllc car rogcrhcr rvith polymyxn B, tbere is a dramaric
reduction of cochlear hair-cell damage as compared i.r adminisrrarion of poly
4. !-oslbm,vcin does nor protecr animals against cis platinum induccd cn
reropathy or peripheral ncurotoxicit)', ' rvhich also indicates a tissue specilic
2,18
C. BUHRER EI aL
Thc molccular basis {or nephroprotecri.rn and oroprotection bv tbslomycin
rcniains rarhcr clusile. Fodblnvcin has becn rcpo|ted to inclease lysosomäl
mernbrane inregritv of rat tubulc kidn€r cells.ri Competiri\c inhibrtion ol toxic
agents iniolring reacrive siles in spc.,iii. tissues, or üe fo ration of conplcres
unsurtable 1or bin.ting ro reaclile sjrcs haic been proposed." Clarilicarion of
thc actual mecliaDisms bv which fosfomycin cxcrts its ellect nlLsr as,aiL lurther
Our resuln indicatc differential sensitivitv of cis platinurn ellects to the
action oflbsfornlcin. Killing ofcochlcar hair cells and rapidlv dividing osteosarcoma cells niglrt be a result of tbe diffirrcnt strucrural propenies (r1 the cisplatinum molcclrlc. This is;n accordance
lhc obseNation that tlre toxicit-v
'vith
proäle of the rhird-generarion organoplatinum compouncL cis diammne [(1 1
c.vi,lobutanedicarboxvläto) (2)-O,O'I platnlun (lI) [r:arbopLatin] diflirs lrom
thar ol cis plarinum whereas Lts anticancer acrivitv is quile similar' ,\ltliough
carboplatin is not {tevoi.l of nephrotoxiciry and otoLoxiciry" thesc sictc clTects
afc lcss s€vere and frcqucnt; rnvelosuppression ocr:urs prominentl,v instcad
Funhef e\perimental in\cstigations ilill be ne.cssarv ro elLrcidate llnr intcr
acrion of {os{onvcin and.is pLatinum in tissues ofiarious origin. Tirmor t,vPc
and tine framc should be gn'cn special thought 'rhen controllcd cLDrical rials
invoiving coactministraLion of platinl,m compounds and fosibmvcin afc being
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FOSFOMYCIN DOES NOT IMPAIR OSTEOSARCOMA
XILL
249
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.18
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Drescbler 1\A, trrbanu: NA\l Hign i..qnenc} audi.nerry ir pfospedi\e
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Ra2;d Adttr
$rd xnn\titüar h Ch;itr lrlthn
li, l99t