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`International coverage of all aspects of phannacology
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`'.'.
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`e Ootrimazole and L-type
`Ca1 + channels
`• Novel CGRP antagonist
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`BRITISH JOURNAL OF PHARMACOLOGY
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`

`

`British Journal of
`Pharmacology
`
`VO LUME 129 (3)
`
`FEBRUARY 2000
`
`SPECIAL REPORT
`'oda. A.A. Selyanko, I.C. Wood. F.C. Abogadie
`J.K. Hadley. M.
`& D.A. Bro1rn D1ffcren1ial 1etrac1hylammonium sensi1ivity of
`KCNQI - 4 potassium channels
`413
`
`M.H. Hawthorn. C.R. Chapple, M. Cock & R. Cht-ss-Wllliams
`Urothelium-derived inh1b1tol) factor(s) innuenccs on dctrusor muscle
`con1ractility in ••itro
`416
`
`H. Doods, G. Hallcrmaycr, D. Wu, M. Enlzeroth, K. Rudolf, W. Engel &
`IV. Eberlein Pharmacological profile of Bl BN4096BS.
`the first
`sclec1ivc small molecule CGRP antagonist
`420
`
`PAPERS
`M. Ushio-Fukai. H. Yamamo10. K. Toyofuku. J . Nishimura, K. Hirano &
`H. Kanaide Changes in the cyrosolic Ca2 ... concentration and Ca2 • -
`sensitivity of 1he contractile apparatus during angio1ensin II-induced
`desensitization in the rabbi1 femoral artery
`425
`
`ishimura, K. l-lirano & H . Kanaide
`M. Ushio-Fukai. H. Yanrnmo10. J .
`The mechanism of the decrease in cytosolic Cal+ concentrations
`induced by angiotcnsin 11 in 1he high K +-depolarized rabbit femoral
`anery
`437
`
`R. Gcrmack, A. S tarzec & G.Y. Perret Regulation of (J1- and /Jy
`adrenergic agonist-stimulaicd lipolytic response in hyperthyroid and
`448
`hypo1hyroid ra1 white adipocytcs
`
`D.T. Th11ai1es. L. Bastcrficld. P.;\I.J . l\lcClea,·e. S.M. Carter &
`N.L.
`immons Gamma-aminobu1yric acid (GABA) 1ransport across
`human in1es1inal cpi1helial (Caco-2) cell monolayer,
`457
`
`L.D. Jayanthi. J .J . Wilson, J . Montah·o & L.J. Defelice Differential
`n:gula1ion of mammalian brain- pecific praline 1ransporter by calcium
`and calc,um-dcpcndent prote111 kmases
`465
`
`C. Petrucci. D. C"cn ia. I\ I. lluni. C. Biondi & P. Bagnoli Soma1osta1in(cid:173)
`induced control of C}tosolic free calcium 111 p11uiiar> tumour cells 471
`
`N. crko1·a. L. Lill. D. Lcibfrilz. 8. l-lnuscn. R.E. Morris, T.L. J ames,
`L.Z. Benet &
`• Chrislians The novel immunm,u1>prcssan1 SDZ- RAD
`protec1s rat brain slices from cyclosporine-induced reduction of high(cid:173)
`energy phosphates
`485
`
`G. D'Agostino. i\1.L. Bologncsi. A. Lucchelli, D. Vicini. 8. Balestra,
`V. pclla. C. Melchiorre & M. Tonini Prejunctional mu carinic
`mhibuory conirol of acc1ylcholine release in 1he human isolated
`dc1rusor: involvement of 1hc M, recep1or sub1ype
`493
`
`ocon1rac11lccffec1 for 5-
`I. Bouchelc1, B. Case, A. Oli1•ier & E. Humcl
`HT,o and 5-HT,F receptor agonists 111 human and bovine cerebral
`arteries: similari1y with human corona., arter>
`501
`
`.A. Stall" ood & D.J . Crankshaw Effec1s of some
`L. Oliveira,
`isoprostancs on the human umbilical ancry i11 ••itro
`509
`
`. Oika11a. A. Murakami &
`. Hirasawa. Y. Kobayashi.
`M. S hiraishi.
`K. Ohuchi Participatio n of m11ogcn•acl1\'ated protein kinase 111
`thapsigargin- and T PA-mduced h1s1um111e produc1ion
`111 munnc
`macrophage RAW 264. 7 cells
`515
`
`P.J . Turner, J .W. Dear & J .C. Foreman
`ln,olvcmcnl of kmm, 111
`hyperrcsponsiveness induced by pla1ele1 acti"aung fac1or in 1he human
`525
`nasal airway
`
`R. Lever. J.R.S. Houl1 & C.P. Page The effects of heparin and rela1ed
`molecules upon 1he adhesion of human pol> morphonuclcar leucocytes
`10 vascular endothelium i11 ,,11,0
`533
`
`L. Dawson, A. Chadha, M. Mcgalou & S. Oury The group II
`mc1abo1ropic glutamate receptor agon,st. DCG-IV. :1llevia1es akincsia
`following intranigral or intraventricularadminis1ra1ion in the reserpine(cid:173)
`treated rat
`54 1
`
`I.M. Fearon. S.G. Ball & C. Peers Clo1rimazolc inhibits 1hc
`rccombinanl human cardiac L-type Cal+ channel 2,c subuni1
`547
`
`L. Shaw, i\l.J. Taggart & C. Austin Mechanisms of 17 /J-oes1radiol
`555
`induced vasodilatation in isola1ed pressurized rat small artenes
`
`K. Zacharowski, D. chncidmiiller, W. lbc. T. GroOer. 1\1. Bucrke.
`J . Meyer & H. Darius Effects of local delivery of trap1dil on neomtima
`566
`formation in a rabbit angioplas1y model
`
`M.A. Yicirn-Coclho & P. Soares-du-Silva On1ogcnic aspec1s of D1
`receptor coupling 10 G proteins and regulation of rat jejuna I a ' . K •
`ATPasc activuy and electrolyte transport
`573
`
`M. Miele, M.A. Mura, P. Enrico, G. Esposito, P .A. erra, R. i\ligh<li.
`I.S. Dcsolc On 1he mechanism of d(cid:173)
`D. Zangani, E. Miele &
`amphetaminc-induccd changes in glutamate. ascorbic acid and uric acid
`582
`release in 1he stria1um of freely movtng ra1s
`
`C.L. Chan, R.L. Jones & H.Y.A. Lau Charac1eri1a1ion of prostanoid
`n.-ccptors mediating inhibi1ion of his1aminc release from an1i-lgE(cid:173)
`nc11vatcd nu peritoneal mast cells
`589
`
`I. Riner, J . Fiir>t,
`A. chmarda, P. Dinkhauser. M. Gschwcntner,
`E. Scandclln, E. Woll, A. Laich, H. Rossmann, U, cidler, F, Lang &
`M. Paulmichl The g:istric H.K-ATPasc blocker lansoprazole is an
`598
`inhibitor of chloride channels
`
`Contents continu~ ...
`
`Copyright (r 2000 Macmillan Publishers Ltd
`
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`

`P.S. Lacy, G. Pilkington, R. Hnn1•csokul, H.J . Fish. J.P. Boyl~ &
`H. Thurston Evidence against potassium as an endothehum-den ved
`hypcrpolarizing factor in rat mesenteric small arteries
`605
`A. Huwiler, M. Wartmann, H. ,·an den Bosch_ & J . Pfeilsc~iftcr
`Extracellular nucleotides activate the p38-stres -activa ted protein kinase
`cascade in glomerular mesa ngial cells
`612
`
`K. Bleasby, S. Chauha n & C.D.A. Brown Characterization of Mpp+
`secretion aero human intestinal Caco-2 cell monolayers: role of p_
`glycoprotein and a novel Na + -dependent organic cation transport
`mechanism
`619
`
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`

`British Journal of Pharmacology (2000) 129, 420-423
`
`© 2000 Macmillan Publishers Ltd All rights reserved 0007-1188/ 00 SI 5.00 (I)
`www.nature.conybjp
`
`SPECI AL REPORT
`Pharmacological profile of BIBN4096BS, the first selective small
`molecule CGRP antagonist
`*·'Henri Doods, 'Gerhard Hallermayer, 'Dongmei Wu, 'Michael Entzeroth, 2Klaus Rudolf
`2Wolfl1ard Engel & 1Wolfgang Eberlein
`
`' Biological Research. Boehringer lngelheim Pharma KG. D-88397 Biberach, Germany and 'Chemical Research. Boehringer
`lngclh~im Pharma KG. D-88397 Biberach. Germa ny
`
`Calcnonin gene-related peptide (CG RP) is one of the most potent. end.ogcnous va,odilators known.
`T his peptide is increased during migraine attacks and has been nnphcatcd 111 the pathogcne"s. of
`migrame headache. Here we repo rt on the first small molecule seleCII\C CGRP a111ago111,1:
`Bl B .t096BS. /11 1·11,0. 1h1s compound is extremely potent at primate CG RP receptors exh1bi1111g an
`affinity (K,) for human CGRP receptors of 14.4±6.3 (11= 4) pM . In an i11 riw model. BIB 4~96BS
`111 doses between I and 30 11g kg-• (i.v.) inhibited the effects of CG RP. released by s11mula11on of
`fl rs concluded that
`in marmoset monkey,
`the 1rigeminal ganglion. on facial blood flow
`Bl B 4096BS is a potent and selective CGR P antagonist.
`British Jo,mwl of Pharmacology (2000) 129, 420 423
`Kcy11 ord5: BIB 4096 BS; CG RP-antagonist; migraine
`Abbm i:11ion,: Bl B 4096BS, 1-Piperidmecarboxamide. N-[2·[ [5-amino-1-[ (4-(4-p) ridinyl)- 1-piperazinyl]carbonyl]pentyl]ami(cid:173)
`[R(cid:173)
`(3.5-drbromo-4-hydrox} phenyl)me1hyl]-2-o,oe1hyl]•4·( 1.4-dihydro-2-o,o-3(:!H )-quinawlinyl)•,
`no]-1 ·[
`( R • .s•)j-: cAM P. adenosme 3'5'-cyclic monophosphate: CG RP. calc11on111 gene-related peptide; EDTA.
`-MC. neuroblastoma cell line of human origin:
`eth) le11ediamme1e1raace1ic acid: LDF. laser doppler fl0\1: SK-
`5-HT. 5-hydroxy1ryp1amine
`
`Introduction CG RP. a 37 ammo acid neuropep11de first
`1dent1 ficd 111 198'.! (Amara N al .• 1982) belongs 10 a famil} of
`peptides 111clud111g calci10111n. adrenomedullin and amyhn.
`Localinlllon studies ha,e shown a 11 ide distribution ofCG RP(cid:173)
`likc nnmunoreactl\C ,1ructurcs 111cludmg receptors m the
`penphcr) and central ncrrnll', S) , tcm (Se~ton. 1991: Bram &
`Cambridge. 1996) In man} tissue,. CG RP conta111111g nene,
`arc clo,cl) a\\oc1a1ed 1, ith blood ,e,seb. Ah hough CG RP has
`a number of effect,. its mo,1 pronounced action 1s ,asodtla(cid:173)
`t1on It " one nf the most potent cndogenou, vasodliators
`kno1, n and 11s vasodilator} effects h,I\C been shown 111 a
`n1ric1y of ,e,,ck e.g. CG RP released from sensory fibre,
`ong111a11ng 111 the tngemmal ganglia dilates cerebral ,·essel,
`(Goadsb) & Ed, msson. I 993).
`l\ltgrame headache is thought 10 be as,ociated with dtlatat1on
`of cramal 1cssels and act11"a11011 of the tngemino-vascular
`S),tem (Mo,ko1\112 & Macfarlane. 1993: Moskownz. 1984,
`Goadsb} et al.. I 99 I). In man ,11111ulat1on of the 1rigem111al
`ganglion rc,uhs 111 the release of CG RP (Goad,b) er al., 1988)
`Moreover. durmg a migrame attack levels of CG RP are
`increased (Goadsby £'/ al .. 1990. Galla, er al .. 1995). Thus,
`CG RP ha, been 11nplica1ed 111 the pathogenesis of m,grame
`the CGRP-induced
`inh1b111on ol
`headache. Accord111gl).
`1asod1la1a11on could be e\pected 10 attenuate m1gra111c
`symptom,
`So far. only C-terminal fragments. example gl\en CG RP(8
`37). ha,e been described 10 act as antago111s1s (Chiba er al .. 1989:
`R1s1 et al .. 1998). however the,c compound, are not very potent
`and their peplldic nat ure 1111111 1hc1r u,e. especia lly for 111 1-frn or
`clinical 111ves11gat1ons. Our goal was 10 1dc111ify small molecule
`a111ago1mts for the CG RP receptor and 10 use them 10 assess the
`role of CG RP 111 migraine headache.
`Our high throughput scrcemng led to the identification of
`d1pcp11de-like compo unds that showed weak but uneqm,ocal
`
`•Author for corrc,pondence;
`E-matl hcnndoodsra bc.boehringcr-ingclhc1m.com
`
`inhibi tion of b111d111g 10 the CG RP recepto r. Lead optimiza(cid:173)
`tio n led lo potent CG RP antagonists, the prototype being
`Bl BN4096BS. I-Pipend1necarboxamide, N-[2-[ [5-amino-1-[
`(4-(4·p) nd111yl)- l-p1pcra11ny l]carbonyl]pen1yl]am1110]• I•[ (3.5-
`dibromo . .t . h) dro,ypheny l)methyl)• 2-oxoelh) 1]-4- ( I .4-dihy(cid:173)
`dro-2-oxo-3(2H )-qmnazohn)I)-. [R-(R•.S•)]- (Figure I). The
`present stud} ,1as underta~en 10 e~amine the pharmacological
`profile of BIB 40968 and to 1111es1iga1e its potential as an
`,lllll·llllgra,ne drug.
`
`ln-vnro exper1111e111s B1nd111g stuche, investigating
`1\ lethod~
`CG RP-receptors from human or rat ongm were perfo rmed
`-MC (neurobla,toma cell ltne of human origin) cell
`us111g SK-
`membranes (appro,1matel} 0.175 Mio cells) and rat spleen
`homogenates (approximately 50 11g of pro1e111 per assay). m1.
`hCG RP 11as u,ed a, the rnd1ol11wnd . The 111cuba11on buffer
`contamed (111 mMJ rm 50. aCI 150. \.lgCI, 5 and EDTA I.
`(ethylene d1a111111c 1c1ra-acc1ic acid) pl I 7.4 Membrane
`homogenates \\Cre 111cuba1ed for 180 111111 al room 1empern1ure
`\lllh 50 pM '2' 1-hCG RP and 111creas1ng concen1rn1ions of
`displacmg compound The incubation was 1cnn111a1cd by
`filtra11on through G F B gla,s fibre filters usmg a cell harvester.
`The pro1e111-bound raclioac111 m wa, determined 111 a gamma
`counter The nonspcc1lk bmdn;g ,1.1s defined as rad1oac11111)
`bound 111 the pre,encc of I 1,,1 CG RP. The IC,11 1aluc, 11ere
`obtained by non-lmcar regrcs,1011 anal),IS on the b,1'1S of a one
`bmcltng sne model. A, values l'Cre calculated aceordmg 10 the
`e4 ua11on K, - IC~, I + L, J..'0 , according 10 Cheng & Prusolf
`( 1973). where L and K0 represent the concen1ra 11on and the
`d1>socia11on constant of the raclioligand. The K" was 0.05 nM
`and 0.1:! nM for the human a nd rat CGRP receptor. as
`detcrmmed by sa1 ura11on b1nd111g cxpenment, wnh 0.5-
`500 pM 1211-CG RP
`Antagonism of BI B 4096 BS was determined by measuring
`-MC cells. For cyclic
`the fo rmation of cyclic AMP 111 SK-
`AMP (adenosine 3'5'-cycl1c mo nophosphate) measurements
`-MC cells were incubated with CGR P alone or 111 the
`SK-
`
`

`

`H. Doods el al
`
`Special Report
`
`421
`
`A
`
`_,.
`Flu, ·~l .~.J----
`
`ll.P.
`
`ISO
`
`]
`
`mmHg •:o J
`
`l,DF
`
`·111inc
`t
`f.S
`
`T t:'il COfll llOund
`
`t
`F~
`lllll:S~0961l\ (0.01 mg 1-i; 1l
`
`S min
`
`Hgurc I S1ruc1ural formula of 131BN4096BS.
`
`Conlrol
`
`B
`
`125
`
`0 CGRP ral spleen
`e CGRP SK•N•MC
`0 BIBN 4096 ,., spleen
`(cid:127) BIBN 4096 SK•N-MC
`
`A
`
`100
`
`e> 80
`C: :;;
`C:
`:ii 60
`
`" IC 1 40
`.,
`'.'I-
`
`20
`
`100
`
`" ::, u::
`
`IL
`0
`.J
`0
`C: g
`:ii :c
`.!:
`~
`
`75
`
`50
`
`25
`o~--~---'---------~---
`0,001
`0,003
`0,01
`0,03
`BIBN40968S (mg kg.1)
`
`0
`r--..---.----,---.-----.----.-~..--..---,
`-8
`.14
`·13
`·12
`·11
`·10
`.9
`.7
`log c (competitor) [M)
`
`Figure 3 (A) Rcprcscnl:lll\c rccord111g 10 1Jlu,tratc the change, m
`bl_ood pressure and LDF·Flux fol101\lng electrical 1ngcm111,1I g,1nghon
`st111rnln11on. (ll) Do>e-dcpcndcnt
`inh1b1t1on of the
`facial
`, ~in
`vasodilauon induced by trigcminal ganglion ,t1111ul,1t1on
`in 1he
`marmoset by llll3N4096BS. Daw .ire mean,±, c.mcan
`
`B
`
`100 e CGRP alone
`(cid:127) CGRP • 10nM BIBN 4096
`
`T
`!
`
`= 80 e · . . 60
`
`E
`
`~ .. 40
`
`:.
`<:
`u
`
`20
`
`·11
`
`-10
`
`-8
`-7
`.g
`log c ( CGRP) [M)
`
`-6
`
`.5
`
`Figure 2 IA) Conccntrauon•dcpcndcnt 111h1b111on of BIB 409613S
`and CG RP of the specific b,nd,ng of •~'l·CG RP to membranes
`obtamcd Iron, the human ncurobla,toma c-ell l111c SK- -.\IIC or from
`rat ,pk-en (U) St11n11la11on of 9chc \MP forma11on 111 SK-N-MC
`cell, by C<, RJ> .1lonc and 111 the prc,cncc of 10 nM BIBN~09613S
`Dai.i arc mean\± s.c.mcan
`
`presence of 10 nM BIBN4096BS for 15 min at 37 C. Cyclic
`AMP ,,as e,tractcd with 0. 1 M HCI and de1ermined by
`rad1oimmunoassa}.
`An apparent pKh va lue was calcula1ed by the equation
`pKb: - log[IJ] + log (C R-I) (Schild, 1949). where [BJ is the
`molar concentratio n of the aniagonist and C R is the ratio of
`thc EC'° values in the presence and absence of the antagonist.
`
`In vivo expenme,,1s Marmosets of euher ,c, (JOO 400 g)
`were anaesthetized wi1 h sodium pemobarbuonc (induc1ion
`wi1h 30 mg kg 1
`, 1.p. and maintained ,,1th an infusion of
`6 mg kg I h 1, i.m) Body temperature ,,as kept a1 37 C by
`automatic control of a heating pad The animals ,,ere placed in
`a stereotaxic fra me and a longitudinal incision was made in the
`scalp. A small hole was drilled 111 the skull a nd a h1polar
`electrode (Rhodes SNEX 100) was lo,, ercd. usmg a m,cro(cid:173)
`manipula tor. in the 1rigeminal ganglion
`to 1den11f> bony
`We used a radiographic approach
`landmarks as guide in placing probes 111 the ganglion. An X(cid:173)
`ray was 1aken us111g a Trophy CCX digual X-Ray a ppara1us
`and the Pars pc1rosa was 1den11ficd as guide for placing the
`electrodes. The po,i1ions of 1he electrodes in the tngcminal
`ganglion were checked v1suall> at the end of each e~periment
`following removal of the brain. The trigeminal ganglion was
`stimulated at 10 H1. '.! mA, '.! ms for 30, u\lng a I lugo Sachs
`Elektronik s1imula tor.
`curomuscular blockade. 15 min
`before elcctncal stimulation. was achieved u, ing pancuronium
`• 1.v .. followed by 0.5 mg kg 1
`bromide (ini1ial I mg kg 1
`every hour).
`Microvascular blood now changes in the facial sk111 were
`measured by Laser Doppler flowmetry with a Pen Flux laser
`doppler system (wave leng1h 780 nM: Time Constant 3 s) as
`nux in arbi1rary units a nd expressed as area under the llux(cid:173)
`curve (mm1
`) according to Escott "' al .. 1995.
`Animal were subjec1ed to two or three periods of eleclrical
`stimulations. separa1ed by a 30 min interval. The first
`s1imulation was a control for the subsequent slimulations.
`Test compound~ were administered intravenously 5 min prior
`to the second or third s1imulation.
`
`British Journal of Pharmacology, vol 129 (3)
`
`

`

`422
`
`H. Doods et al
`
`Special Report
`
`Results BIB 4096BS completely inh ibi ted the specific bind(cid:173)
`ing or 11' 1- G RP to SK-
`-MC cells and displayed an affi nit y
`(K,) of 14.4±6.3 pM (11= 4) for the hu man CGRP receptor
`(Figure 2A). The endogenous ligand CG RP itself and the
`pcptidic antago nist CG RP( -37) displayed affi nities (-:,) of
`Jl.7± 1.6 pM (11 = 15) and 3.6±0.7 nM
`(11 = 4). respecti vely.
`Employing the same cell li ne it was shown that Bl B 4096BS is
`a pure antagonist.
`In
`the presence of antagonist
`the
`concentra tion-re ponsc curve of CG RP to increase cyclic
`AMP levels was shifted to the right in a para llel fashio n,
`indicating competi tive antagoni m (Figure 2B). The pK• value
`ca lcu lated at an antaeoni t concentra tion of 10 nM was
`11.0±0.3 (11 = 3). In addi tion. concentrations or BIB 4096B$
`up I
`I 00 JIM did 1101 show any ago nistic effects. The affinity
`(K,) for CG RP binding si te
`in ra t spleen was 3.4 ±0.5 nM
`(11 = 3) and 96±4 pM (11=9) for BIB 4096B and CG RP.
`respectively. Accordi ngly. BIB 4096B$ possessed a 236 fold
`higher affinity for human (SK-
`-MC cells) tha n for rat
`( pleen) receptor .
`Electrical stimulation or
`the
`trigemi nal ganglion
`in
`marmosets caused an ipsilateral increase in facial kin blood
`flow. Following timulation there wa a rapid increase in flow
`which returned to ba e line value after app roximately IO min .
`This response was obtai ned th ree ti mes in the ame animal
`with no significant difference between the firs t and third
`sti mulation. The area under the curve fo r the increase in flux
`ob erved was used to determine the effect of BIB 4096BS
`and
`amounted
`10
`I 12± 11 111 1112• Admi nistration of
`Bl BN-10968
`ignificantly and dose-dependently inhibi ted the
`re ponse e oked by trigeminal ga nglion timulation (Figure 3).
`A 50% inhibition was achieved with a dose of ap proximately
`3 Jig kg- 1 (11 = 4) applied intravenously. An almo t complete
`blockade was observed wi th a doe of 30 pg kg - 1 (i.v.).
`BIB 4096B$ in doe up to I mg kg - 1 (i.v.) had no intrinsic
`cardiovascular effects.
`
`Discussion
`In order to determine the affinity of BIB 40968S
`for human CG RP receptors we decided to u e K-
`-M cell,
`since the recep tor cxpre ed in 1h,s cell line is identical to the
`cloned human GRP receptor ( iyar et al .. 1996). The affinity
`of BIBN4096BS for the human G RP receptor i even higher
`than that of the endogenou
`ligand CGR P itself and 150 fold
`higher compared to the pep1idic antagonist CG RP(S-37).
`Moreover. Bl B 40968S proved to be a competi tive antagoni t
`without any intri nsic agoni tic activity. 8 1 B 4096B$ had no
`ignifican1 affinity for a et of 75 different receptor or enzyme
`system (IC 50 > > 1000 nM) including calcitonin. amylin or
`adrenomedullin receptors (for complete Ii I of a ay , see
`Rudolf er al .. 1997). Therefore. it can be concluded that
`BIB 409613S i a selective G RP antagonist with high affinity.
`We also demonstrated that BIB 4096B$ po sesse a remark-
`
`References
`
`AIYAR, N. RA 'D. K .. ELSHOURBAGY, N.A. 7E G. Z .. ADAMOU,
`J E .. BERGSMA, D.J . & LI , Y. (1996). A D A encoding the
`calcitonin gene-related peptide type I receptor. J. Bwl. Cl,e111 •
`271, 11325- 11329.
`,
`AMARA. S.G .. JONES. V .. RO E ~ELD. M.G., O G, E.S. & EVA S
`R.M. (1982). Alternative R A processing in calcitonin gen~
`expression generates mRNA encoding cli/Terent polypeptide
`products. "'"'"· 298, 240 244.
`BRAIN. S.D. & CAMIJRIDGE, H (1996). Caleilonin Gene- Related
`Peptide: Vasoactivc E/Tcct and Potential Therapeutic Role. Gen.
`Phtirmacol., 27, 607 - 61 I.
`
`British Journal of Pharmacology, vol 129 (3)
`
`able species selecti vity, exhibiting an approximately 200 fold
`higher affi ni ty for primate CGRP receptors compared to rat
`CGRP receptor . The same was ob erved for other non•
`primate species, such as dog, _guinea-pig and rabbit. Having
`established that Bl BN40968S 1s a potent and elective CGRP
`antagoni t we addressed the question whether BIBN4096BS
`has the poten tia l to be an anti-migraine drug. Measuri ng facial
`blood fl ow following antidromic limulation of the trigeminal
`ganglion has been suggested a a model to evaluate drugs that
`target the trigemino-vascular ystem during migraine headache
`(Escott er al., 1995). Antidrom1c stimulation will result in the
`relea e of neuro1 eptides uch a CGRP. Concomitantly, an
`increase in blood n w will be observed in blood vessel
`innervated by sensory nerves. A the facial sk in is innervated
`by the trigeminal ne rve. we measured the blood now in this
`area. Because BIB 40968S has a much higher afli ni ty for
`primate CG RP receptors compared to rat CG RP receptors. we
`used marmo ·ets. BIBN40968S completely
`inhi bited
`the
`neurogenic va odilation. demon trating that CGRP plays a
`major ro le in the vasodilation ob erved followi ng stimula1ion
`of the trigemina l nerve in the marmoset. This i in agreeme111
`with experiments performed in the rat by Escott er al. (1995).
`Although CG RP itself is a trong vasodilator. i.v. administra(cid:173)
`tion of the antagonist had no effect on basal blood pressure
`and heart rate. This might indicate that CG RP plays a minor
`role
`in basal cardiova cular homcosta is under normal
`conditions. and that G RP is only involved in (human)
`pa lhophy iology. Moreover, it i of intere t to mention that in
`our model anti-migraine drug
`like sumatriplan and zolmi(cid:173)
`lriptan, both 5-l-JT 111 ,, agonists. inhibited the increase in facial
`blood now due to lrigeminal 111milation not completely, but
`only approximately seventy per cent (data not shown).
`Accordingly. it could be hypothesized
`that blockade of
`post ynaptically located va cular G RP receptors is a more
`powerful way to auenuate CG RP mediated effects than
`inhibiting the relea e of the peptide by timula ting presynaptic
`5-HT-receptor .
`In conclusion , our result demonstrate Llrnt BIBN4096BS is
`a high affinity and selective G RP antagoni t. Moreover.
`BIB 4096 B
`is a very potent
`inhibitor of neurogcnic
`va odilation .
`ince several line of evidence indicate thal
`CG RP might be a key factor in the ini tiation of