Note: Descriptions are shown in the official language in which they were submitted.
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
IÃ~~trat~~:_~e~ati.ve In_,.:~ in:~~ Methods
FEDERALLY SPONSORED RESEARCH OR ~EVEIyI~PINM.IsINT
11.3.s i.?#v#:fli:Ã.;.3n was mt3do wltl` GL?i'C=rinnent >:.eppo1t under
GF"i1?:t Nf},RW'CA-
, ,15296 1d~i SaÃ;dZ`d by the Nil.at'ÃL3riai In5:Ãi.l:~tes oziHmhi:3 s?nwa
<_?rai?! No. DT_.TGt,MIER6,31 Ks'
~.fE'f=~' }', Tbe Goi'L`TTixilw.nt it%hs e.'.eÃai'3 rf t,i33., ?,,3 the
`IEC'~~~ICA~ ~IEf"J-)
T.i.iv ?:1'4 +? .lt? 3 TS :hate4 to :7C'fl'3`.~dS aS.?F.'
:d7t7"<.?3peF`23.ik.Y=?Q i721c'd,~ll:g `i1s:Ilg- iiui;?`t 4uii7;
i:oÃ3"apC?Li#: dti, compounds i-hat fl`'r.?C3TosC.'.. in the ?f11'As3bzo
i.ight (;.L) region e-a :aiv
s,pu+.:i:?'utn.?.L., above 670 Fil'ia.
1 BACKGROUND
L:~in~ surgical procedures, it is often difficult: li3r i:~ie, ~"'A; g"cota :o
t(l~.~
location of s=Ãua.airws stÃ.t:b as the thoracic d'ltcÃ., UIii.3ar?^ tr:act,
wid b~~dli i.t.`v' trw:., and
_?%'. di3ac>ting i'ar:er"3;.:s.
SUNIMARY
1w ~i~ .?.. ~,,, ',..aa`.~., :II~'te`~"~'~is based, at least 1"i ~)w~l~.,
:?i~. ..+~~. discovery ~:e~~;Ã.it ~,-:.'ÃFÃ ~~.Z'i'S
_ c~,,.. t1~,.?k sahc3t $euo1esou~ in the inv`.s-1blE: light (IL) i'C'.~1C?k',
of the Sr7tw4'~?~i:~.aii are t~4G,~i~.l ~:3:~ ?Y;~.?~?4 3.Ã3;.,M
oc,t:in s nbEtoliiic:>i structures, e.w;., during surgery. Thus, pr?v1u.t"S:1
6w,.eI`a, a:'e mehoCi4 tbr
rF:'1 -.n:y sta udl-u.`es sLit h as td:i fl>mil~. .4c duT.t. urinary iTaciy
J13.Ãc2r-y tr#J:,., and i3er..:3i t3tiig
arw 1c:', using IL LaS'es,20 3ia : fi;:,f. C]k?~,'~:5:t,
3.S~k~'i.:Yk''e?i~:1.iL,'~.~ features 2.etLkSJ(5s AGI, ~~~.~2c~~ Ã~>~k:, an
c.?~1~tÃ.tt~~rci~.-'C~i
a
S':ruL.tI::`c seie'~:iS..:i fs"'ozTi the gmi,3p ,f,'..oI353.sfiASa.~,~' of a
b}.3.:<iÃy, itree, urE;1:s:.1, l.1."::3 p3.:i'L7in i ae:':."`of:
ayl v.?v(`. >71w me;ht)ds ?a~i a?li~~ zlf~ll%13~i4~~=? Ã1~S' to a s:fb~ecÃ. a
pI'cp:?`c#.t.Wi. ?ziwlWm,3.', :3r,
zn~.tsf~.'33~:< lÃJm` (IL) fl'L?o# L?p`,.i'e that Ã4 fiftE'red b''fh~.~
kIdnu~% i,it+;..3 t~iL urine sÃ_~. ? ~'...[:5r.:t~'
by the ;.~vcx ixa<r the b-iIY;:. SÃ>iai3., or both, and obÃ~aÃntng an ix~~ ge
t?_i`1L wz3t e'len',
2 5 tallas.lor7's z:i"f:Il the fil?.1or#3;3hC3xep, whGS4,1G:1 the ffla<f#?G;
is a iep1"esC;?att's:txiEf?;. of the aÃ~i:tl.oml+..al
i-mclL?Ã`S..a.
In s, i,?dm 2aspC.ct. the itÃ4S::i7tRC?tl features ~I~.offii.3d.' for
ii`Iiak;n~`..~. z3: thLS,caclc da~t or i.;
pZS ~ion tÃ.eim?f :n V'?vE3. '.f'Ai~E.'`~ m~.''`{thods Ãns?lud5v iqjt:cting,
a prepa.atZa`3i.i a.{-iC.'ll~?~E#?i;:'i;?>
w.
H ,a:~.liL?or:?phoi'i: (11.:F) Ãnto a lymph I%t7[ac t?fa sll~#f'"1' SEIC}i
fl3:3i. ?t <?ppZ.w.i 5 in the .ht3i'::cii;:
i
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
duct, e=g, any aymp;3. ttodc, tli at drains into tl-ic Ãhc.2-tacic duÃ:t, c
g., at or t~~=}s; '~`,~s level of
the di.a~ph?'ugprn; und obtaining an 1magi;~ of 1:ivis1blL z!ghia (aL.)
wavelength cnriission:a:
= y. = .. . duct. :: ~. Ã~i~taÃ,~e is a i'~. pÃ"Gs~'Inabl~111.In of tlc
iZ~iT~i~.i?;~. i.iÃ.3t.t. In st 1">i1e Lt.1bo-&Ã1?,ents= a
8,:.1ffi?:=lvÃat w.nnt#uÃ'~t~. oftime is allowed to pass ior the ILF .Sr flow
into itiL tAi<)ras:ic i.-tÃct
`? ;lcft;;'o t.:u Ãmagc is (?llÃ<i:zlle;d.
~~ ar < a.a 'v t7~ ~ ~ y r c~'tl ~s
i: :~+-,F?~S`.' ~.eia7f.9{~7Ãn?L.II~~, ".~li.= 1T?.c3.~y:. ]I34:lui~e'.~
;iS,Ã31~: ~ZZ3~2'~aE?3~ Uf ,:?a: S?ajv~:i x... and .C~ a:3e
lynxagC'% ial<...,ide's x, fir?it ii-nagv L?'..stiz3.Ãned ftGfl';r Cfilc or
3IiL?"s('_= wavelt:.=nA?t$':3 of 'v'24..?3e light and
..
soci?ind i23 riga~ i}ytai;net3 fiom. IL w:t oliingf13 emissions of the ILF. hl
4:?mo eIi3,bodimi=s -1is',
~~1~. a_I~i:~
t^ 5~~ ~= ~t visible t t the ~ ?i3f',=~ ".,~Ã,Ã`~:~~' a Y~ I'~1~3i'~'. light
image of t~1C. surgical field a#';I.`~+. m 11, w=`'vC`A'a:21gfq IÃ;:i~.g-4
of .,.3e s u, m~,c:3l 7::',.id. hi 5i?i.li einb(?Ci2'Ãi"3:a`:IIts,, fllU F'i
ilb I c :igh, l i mage and :}':Ca. .`L wzxv +`.',o:ng:h
i:~~afso are obtained L=oncw>ently,
In `mti?me ei3:bodIi3:'La,iit:^`., 41E?4c .n.Ãrs.'~.g an 1.nnage
Ãili,'.32.Ãdes positioning an 4?3::<:brC}ilii';~
v
i;.7? Ã?y.l~ j . as i;'3i;.I?:~tVd ~'i~'.Ã'i::4:~15 %'t~.~i3~:=~:Ã.i.a. to
the 4GÃ'I~~.E~,.''L`cF'~:~; 4#t..iÃ:=T' `:...;~:, structure or
~:32ic S c?eo Y`if.`i~ ] `v'ai> \ ~ l ,~_~)~)ed. so ~..`~``12~~. the
`~~?v _aww are ?n c tJlc enough prÃ3:S1mEE-y :o cIIt:bla:3 tilt;<
In so:Ã1i.o cmbC L>i>i2t;:its, the preparation is i:idx.l;;ili s14'i'ed
systeib3ica.l"4', e.g.,
iii,T, ~'G.'~,LZ uror y, ois3ltt:mi,Ci.velya by di1~ot injC:[,'.t3.t3E1. The
<?da7iin#?it?'afi43Ã. is not t.tip2cad, In
some i'irlboC,i.fi.nez3t::, thu an~to.I,~dicii~ structure i~`s a bÃfit3"'v
tree, and the pà a;~.3 t :a?:ioll ,s
adk:tf.1=steiii:$ by .i2,S ~ .1i3F1 (c.g, caaÃni.3lati#in) into a f s`3.,i.a?Z
vein, 1~:ft >iepi2t:Ã:. &,Ic$, or Ã`i:3t
r;4 h>..pcnii~ duC.i, hn sonni< vi.:.nllf-r`s~s'e$.mo,~'.i#S, tlIcaTi?
IfC1:'I724a?r. s:T`E?mit'8: I, 22.aIa~:~t.L~, Ã#'1#r 4.Enw
;`fÃ=ep<!i'aEioa.. is administered by iiWa>tIon into a renal artery,
bl:ÃdS=C:1`, fi-r In se,.Ã-nt;
~ ~d~lieae'r C lÃ~..II if.ei', t`
ennbLiYIÃ;:il(+I`?t.~~.' the ~,=nC:t.3ods IInS: 3,Ão~f:
'ct'L1i.:nillEsii[3t1f}I~ ~.Yl`ie ~ ' `~~ ,, .
aceÃazo`i:mid2; or sQ~churn, bicar;sonatc.
i:i some emlbodÃmonts, the ILF is a ncu-ini:rarecl fluorophore (NI~.~.a )
wi~~~i ar?
25 `'..:T3is4i?'o?n :i avt:iC'=ingth in a T<3a3g'l.' frE.i2'3'#. about 670 F?m
to about 1,000 .`3n,,, t:;.g.s, .tRDyC1 8,
:RIDyu~~~ jRDvu'38, xRI3ve4(,), J-R.DyU4l,. IRD-ye700, HRDvcSOO. Cv5.5, :un:
Cy7 r or an
au,u:log the-mof
In Sorn enlbodtmctat,~, t'cie Ticar-inftdÃred flfac?r~~p-hore is nneth lf tsl
2ie (_~-IB'~, h.
Yi.37?F: c.mbota?fnuÃaa:'õ'~~c: p#'opai'at'i.oai is a solution compt'iS12ig
w:.~oÃlt :1110Ãi'A .... 14,`S, +-&I N1:s.
iQ 3in V:E;frini`.= embodiments, t1h(` methods InclÃ1de c1d3nI#niS.f3i'iing_.
,_2 ,7siÃa;.nft r.ÃiIn `iz- ?; MB tci.
achieve a cor:cs.ÃnfAafic,2i t?f abota~ 10 --- 40 ~tM MB in the ti> be .` 4_d,
nn, aon-w
;'
E~:Ãni'bt3i~ÃI,?.;ntsY >1'n~. MB is ~.C~~7iE>11S'~i.'i~:d in a if)f~~Ã~.
Sl'St~:.Ãn"Ii>i: ~`~.3s~, s?s a:i3'v`ut 0.1 w :i; mL?'r>.`r.:?f
bod.f+wt;igh?.. _;I3 :ii;i3ie i:Y?`#bo#.lb3'iz..e23.ts, the M13 i,;
~?dindaÃ4t:i:=rC~S.~ Ã.3 a toUl s~y)t(=.i'?1e-c da3so f}k:
2
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
ci;ii.,?ut I m,:/kn, of body weit l3t, Id3 stI?ne embodÃI7te:`Iis, 1:iiL' MB
is admin;:JteÃ;wd ir, a tE_tva>
Syq-e.Ã"f.'k E:t?se of less than 1``~ ~ng;"kg, e,g., less than 7,5 m`4-'1kg,
of body we1ghi:
In a.ilota?C;Ã" a4peofs? the invention provides methods for imaging an
al1a`eÃx3.(:,i:l
_ ,.f v=
Js~`Ll~'.t~~.~`4~p ~v'.W~.} vasculature, ~~i~~L~.~~~ tree, t~~.~~rC.~.i.~~.Gs
duf:l:. ureters, ~~,'L,'=C~d.i ii~~`ci~L~..~~u~.~%>, w
V~ V = Y~ri V cells), ~ a5 ~~~~ .~if, ]
i~ ~~.3ext'ta.~l~?~"L i~ .f~.~~i+a, a ~';z3~E~~'('.='cl~r 3rslÃ:~, ..~..a tu
mor `~...~.., t~6Ã?>~s.f'ti {t~' ~}t3~?t ~ Ãt~ a. t)
parav}:yroid origlai;, or a portion thoreoi', in vivo. 17he rue.Ãhod,--
include systam;caI1y
"1Ã tY'i? t'Un1'3ilsly; t`.idM;I31St4'Ã"`:1g to a. Si3~:Jcct a preparation
Ix3ebutiÃÃag fn+`,'.thZ%lL~=ne 37lE~'L'.
w:Ãch that it appears in ( .c. fl:rL'S'S #Ilto, R .f ., by roÃ3`~1gi lt~e or
afl:ert?~~~Ã:iC flow) il'ie
am$.a?i~.~.~.ÃLid `t'~".~Ltule; wnd i?bia.n1n''~~`, i3.i11miÃ~^;tl
i,fz11.1f:3n'.-slrent %'.nn14sii)Ã3s I3"i3"i. .h`v' N1.13õ
`.i' ?iÃ'.Ãn i ~' Z Ã4 ~ 3'r,f'1t:i2~~.~:iC1~, i;~~: t}l``.
~.21s~ro.i~la1Ã`~:~%tl S~i~3~::i.`,.Ir~":. ln:'i3a~:tz :~:i
of ~ `s Ã~` , r , , ~
~ t'~~sli~'~3:#
~'w c`>i' S3~'M,~;t.~~x:` uptake C#: ~=IB, `'~`Ch ~Ã.;~ hi.'~:lÃ"~.
~l~"k"+~`~ea~.Sr.it~..F, i3i t `~t. p
gdti.
I:El a f1~thea' as~..tect, th`a? :_1:Ibri .aitlof3 p:C1S..`ldk:..a
rF.ieffiods~:s~> Ãas3u?>il~
aruxwt-nieal~ s;nucÃurosa.t vivo. `~~-ic =-i-ictli~.~ds .ittclLtdc
adÃninister,ng, e.g., s; r.b~.,m~.call
i4 sI'"111#?i3si.i:3i7Ã4: (C.#.;., intravenously), to a sllbjf:c~ a
pR't:paF=7.tki3 ~I3 =~~~~~~~Ã~? :i~:: 3~~`~ ~r L bi:.Ãc
M,~~~ such that it i3~3pi',a: i in (i.e., flows id7R)., t:>f`.v., by s
C.trotMÃu.deor a.iltei"45~?,~ Ãd+.~ f:+ E2 ) i.;:;sv
fiõi` anatomical ~trÃ:I?wfli$'~,', C3~.E7ti1ÃnF,ng a flIst image of
iÃlb'A53:"9lC: l:gl1t wavf3leiiL ,>? ~:lllf~~lt:> 1
t?.i `...7e ?1013, iFihe.'e:Y2 the image is Y iZ:.paC'i4=ntUti4Zn of t}2e.
first a.`Ã:`att)ÃIlical stF"`L3.cÃur':,
i1d-m:nÃsteaing :~.~ fluC`= ::1l.b;+e'Sct a preparation including a second
la]v14;blC lkg,11112i :uor~~+`pho:e
#~.~,a` s~.~i~n 3vtba~,it at least about r;` 8~~ ~?Ã'.~~.~i~, i ~ ~.~ 't .~ir
Ãt-~? +3vv ., '~~:
s.~L~ ~.4.) with C~+?~~..ÃF~ ~~rtt~i~e=.:~lS~1 1 of Y#' l s.~.L ~.., ~ ~.6 Ã~
. suC.l:.
~ x ~ i
¾~~. s~.'~;L~.l>:i.~ :LI' ~:~t ?~}~.::i:i'.'s in (Ã.i~., flows into, 4~.g., by
i~3tA'(?4~':~:Ãii:~ or <~2t~.~"~.~gÃ".`~s.a:Ã~: ~.ii~L3 t ~:i'..e
second an.ÃtL.f2n.t.:=Ãl structure; and f~~~tai:i.ÃÃ.i~t'G..,. i.'Ã cseconLf-
a~mag4 of invisible i.igFi.eeÃaõÃ~"~i>"e.`ir:~
c-
+:?f the second Tc :' ~b'}lE r`~:3a3. ~.?7te image of the invisible lI~~ it
emissions t3~3~t~1L.~ St;3..a 3.~.=F is
art'.p;eSenE.a'liii?;., of a su~.:oÃid anatomical structure.
25 In Na9m+.''.1 embC?d1~1^i4Ã1~5y lÃ~' anatomical xtrLZC tL:3 : rfi.p`t
.Jf..31t~:i3~. by ~.~.3:'.` ~~I ~t a"~l ~ xo i s a
pÃ3rof3n,ui the a biliary tree, a thoracic duct, a 1.3.Ã`.LtC.i. ""s~>`~..'
beaà i a pjrat.,.~'roiia
.O..+~'.,~.nfa_. a pancreas islet cells), a tumor (e.g., a tL1moÃ' of
pancreatic oi paÃ":#:t.~~-vmÃ.d
t3~~~~ ~ 3> +_ F' a r3sw}: lÃi,Ã,. ths.Ã`i3iÃa:, In some :3"~iW1meilt'd, the
taÃ1ek-an1ccl:z .e~ro seÃ;ied
bJ the 5coond image 15 i.1 poIliol'': of the vasculature, e.g,: bli2ar"y
t:"ea, a f1h43racic duE:;'t, a
30; Ute.el', a .wa`t. 'a pwrckth~`2'otd ,s~~'i~3d, a pancreas (< .~.~., islet
cells), a tw:3'if?F' a t,?ma:r of
kZ:.ancroatc o;" paÃ'%~~E~oyÃ'{"ai3 ongÃ.Ãi)w or :3 poi'tio:> ffiert,:olc and
is dÃfauLnt from iE.':t: aÃ':atc1l-nic:::.ll
s2'rucLuL-.e .'~iV`txie: Vi,Zted bv the IfirF.,~>t :ÃnuC`.Ãge-
3
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
Zj.i sorS.2e. eS?..ibS.+kLALkS.eR?t.3. LASW sS.i1.ro.li-iSS ILF is tF.blL\.!c-
T aa;ine gS,Ve3.C $ }CG) G#.Y!L, ci rboxZtli3..
acid k>f I1~Dye'11800Mz
I:'1 sS3Y?a:`' ~.=Ã":?bodÃ.3?Ln:s, ta?e methods fuit;.e:' include obtcÃi.-d-
u~ ..? visible light ia:::E~;;:
~ all of ~.:3e structures. The .L.ti4.i,~.lla;gt:, ueG;f).3E~?8Y7.I?:~ix and
LÃsa:~.14 light ;:lt~.ag~`:, cc..,: :Ã~i> bo
obtalaI,~:~l co.?.eur~ent;y, ~:.~d opÃiormlly superimposed to crea~o a merged
im<?go;
in i:Fa addf}lz?nal i?7;?t..i. the T3?ve?1t1ixi features "11Ã::hodsiC1> in3
iginf.~, e?. A7t3::"t.f;I"I of ~.2~;.;;
'~"i in ' Y1 ~~ 3~ F Y' 7tt ~ a 4 Y ~
t`~?tic4~lw`:.~arv Ã32 ; io L: A hrv> s~:vi.`L~~'fS a:~~'=1 aZ?C=.
<3~.~.~?~.fa~li',t~.~.`g t1~2`F.~~-e,.A..~j;:t i a pn5,`.p.'d.:ati.:nciud:ng
met;?yhmeb1ue (MB) such t~iat it appcars in (Ãx,< flows, :l?ta, O.g'~ b;
.'rwtr,?~rad4
or ani~ro:~~ade f.ow) :h. vasculature, ati.i obtkainiÃ?g ai:. imag;: of
iÃrv;siLsu: light (.;'L
;:,t w..4 eiL-'ngffi i 3?'i~~~?CaÃ? u`bLiciia iffic iit>is .Ã
Ã`ea31i:se?]taii.Ão.s.]: of ÃA7e Z <l"c?:.: ali.?m
..i:~` .~.~'~.{~i ~r.~] a
I_,` a 2 ?.aa~s~~ t'~~ ?s7 ~C~~, a~1~L Ã.Ã "E >~'e]]tit3Ã? ?~i+~ ~' ~Ãi ~
l7Ã3Ei4 s:a ~:3Ã FI?~%.Ã ~ zTla ~', ' i1Ã# vc .\ N
. k.~ i ~; I"13i~
ii.4wiu ~~~~ ovG:huat>nn of ~lzip design desirability, assessiner~t of flap
,riabffi>;, ,,;-
it_;:w>.mtÃ::%iEoi? i?Ã ;^uÃtt3b.2hi'y of a Il.i3i3.d'.ng.flCip for salvage
therapy wft2l l..ÃV'i.~zi?<AY".1cs. Tf:i:
methods ?.i?;lud+`,:: ad7?37n?stiE:#i21M to the subject a preparation 1
.?Gli?~~:1rF: an invi:iibic iig?Y.i i L:
fluorophore 3ui:h flhat Ãt appears Ãkl flows 1:3'Ãfi0, t:=,g., by
riftoi2Tc:iCae u <331ÃE'3"4?~.,ryzde fl+?v,`)
ffic v~^ws:'. 3iz iuf'e of flhc tsSue, aFid obta1nÃ4't- ag"3 ]I? i9,.xe of
t;.?'v is"INQ lT:4:~.it w:3.vGlengti7.
a::di;l-s:'s1L`}Fis, w:ioi'..Rn the f?'la4se is a rep"e4eI?.Ãat3'l?.Ã"~~. of
the vwsC:u.iatilri:, of the fi~~ ue. 3nd.
wherein \l1G vf=swulat:m~ of t};utisSL,e i1:d1ct3.c;~ the des?Ã'a:3-flTty of
the fli:Ãp design, the
~~labilztyof ta3c flap, oz dic sultabiii.yofa tailÃp~g fiap aor ;~~~lvage
therapy wi a
An lÃgr?t fluoroj5horC (IIJF) is a compound that e:Ãi't4 Iigj?Ã in E ac
.i i~. ; `
~ , of :h~' `~+uctaL7~?? (670 I?I?'z ~i? 1 ~~~~ i~00 1`f'~7) such ^ ~ ~ c 4
F:t~=eEe z3.1:I"s a C.A
:i3i'4'i,i.~3~~; ~Ã~ ~?E.Ã ; ,.+
a?F:l ti` 1000 r`.~.Ã11) to rnit infrared (1000 mn tZ) 20,000 imn) to far
infrared (20,00~~? iun: to
>.00,0V02i.2ri),\ -s any light (A~~v4= 67\11a31Lf is iIvR:3tb.(e to tj,?4=
.Ã3.Zdk.Glk human \+j Yr =~~Iw.7~~v
a;? :: ?vISible lÃgght. f1uo?"oph+.?I'Z:s {l3 not s6bsai$?'Ã'1"w'lly
0.=l?aaigG ti1e appuI'ei:i;c ci3AÃ?3o su#'g~i,:.a:
field, and because tissue aItoflE1:"LsGCÃll:eilt Ãz?[~~'wi?.iotei14:¾h;d FS
oTi%, detection
. # L S-~} q?.W~Ã`[)~?3.?tZ~~~':~
l's`~~a ~a.Ã~?3_7~~.' %?~~~.i.t3 i~.{s`~w ideal reagents 4.f~~ `.~'{.~Ãr~;?
.#.~ .;s:;
.,%'?7e3"ia:g. 3Ã1 sÃ)an'L: en?bodÃmonnt ILFs can also include
iluZ7rFphÃ3:"es that aie. v.Ã~,,Ãbl`v :o, z;,#a~
Il?keZ cZ i z?s >t9;?~,=~ i34 they ;1ls:?fluibÃ'cscG`= in the li?v2ssIt3la.
light I`f:ÃJIi7Eb.
'>h, iurf.3 "near Ãnfz'arCd fluL`_ti'ophC?re'i' rtaf"e?'`',~` tt? ;.}at-
1pt3?..Ield:5'.hcl flzfLL.3..:3f:t:-ndl the
a .5,,.`s. d :nf1a.e\f. i~eg,?on of th-c spet:i.I'ui-n.. i.e., about 680 nin
to 10t.3~.i nÃi.?, jI'l.3esC l.#F ~L ~Ã,~.f =".`i
incl.Ãdo ind.~cvaniuc ~Pc:4~ (ICG)s IRi)ye~'~,178 (Li-COR, Lincoln, NE),
1IRDve'? O,
IRDye3g, P-,',:Dv~.~0 :[R:~ve4 1. 1:RDyc7 00,1~yc"RASOt#f~~' z;L1M COIZ,
mi'ct.a:d to, ut:.:ziw a.s
4
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
CW800 of CW800-CA= ax,, th~.' cw;bt~xyliU acid form~ of the "~~R flu<>rophore
~~~DyJ~ ~04 C,115.5; Cy^?'. Cy:5, 1R-786,1_)RAQ5N'O Ã~i N-oxideniodit.~od
abr:;q13im3?:`4'i$ quantum dots, and ia.7i.~.lo~.'.J ta;iluGof, ovK'.
.}`,}'d;C.sph#i:Ec. ;.analo4 S Z::-
ti;:I~p~~o:~i3.tLi3 analogs t~7~.~'Q~_ .II'1 some ~;I7i~?[~t~1Ft-1,ill:~+,
tli~: ~''~.I~L ~t is a i:~.I~Zri?pt3t?z e i.Ee:zkà sw;d
15 in US: P?iri'i;,i,crna~ ~latUrÃApplac tr~.~n No. 60'835,3144.
T ha: ,,r~iae5ori methods h<ave a Iiti13'Ibeà of advantages. FE?: e;$S"<'3.
npzet. tl1'~.~. :yC.s usodi
l:exein can ~~ iYZi`~~~~~% ~? t~ *5' ~ z3i s3~~ ~ 1;11tCXl~a 3~1f3T1S, yet
it:'e!A.fl ab.s`te ii3 ovadc-t:1a.elleIlt
4J:s:.ai'rlut.o,3; For e-munple, fl1c doses i3finCthylenc-:.~luE~ (MB) used à -
ki tffic i~.:z3re`cL.ia:i;
~,Y..h.L,t,~~ .o h; described `i herein are ~z s2%`~D`illai~YtG~1;~l~' ~
C=tiÃ..b 'i~~.;ta ~'.t}~=~~~1 ~~]ia~z :~~~~:i+., ~3~:~.(~~=.x s.t1i. MB i.S
:t ~ a r > Ly a-+ toxicity ~ ~s;SCr t
used w:.e a ~:"$,~r~:~? ~~~#.~L.~ i~.~ t E,Y thus avoiding E~t f,'if 1
~T~$i~<:' ...~2 .Fl~. tf>\~4~ `c.LT.=xe;iti'.u W IS.h
MB. Surprisingly, even at these low d<,3scs, MB is seorC;ted or partition prt
te.eF'.fi3.illy i;'11to
ur'lz3i:`.= zw>d .33.aw; :a4 concentrations thuit.a~e optimal JtC?r
xli3C3.a<3G3: .n: ;'isua,1zatiCsf, :a:IoI`ax?;h 4ui:=a?
%tSnE:`.=vnt1". .2i~ms (+.:.ti.} a c+?I?cUnt.ra:El.o1a. in the tTskLii: of
only about 10.r40 ;i.M; cg,A about 2S,i
a~~ , M) are far below those useÃti` abr ~~ipual Ji;;lit imtaging:
defined, "~ 1). a`.it"`~7's S."tx1+~.;> yxi..4i; izall tc5:<h.l.c:tl a12d
s!i.:iE''T'zbfii= :E.n3x.'~~. :.;\f.4,t hh=xLir, x*3as L the
~:ii~:t= mi;~:iiis,t.~.~,, ws ct:rn-mt?A l4' un(iS:I'std>i9t ~. by o"tio of
ordinary skat: i1~ :?ti =:trt ._? .4'h14.h th1.3
i?3v:#'i.lot: 3Te:.oras. Methods and, materials are dL'~`.~'cribC:d hGTGa.I1
for E_ a,: in :ali: pzc~4i.:i:
;n:t'emua'S11 other, suitable mo-r1CitiS am~d materials kn<#w-?t 1,`. -ffiL
art can also be usod. Fhe
materials, mefliods, and exii1n~`+~~s:s :~.fL 1~:lL~~.~!"i.3t~:~`c% Oiiz~.
w1x not iiitt:i.i,`:~.l t~? be d,:l'.`.ttlll4, s ~8i
applications, i.~c~teÃ1tS, s"~';qL7E.'nC:e47 daia`?.tJ: entries, zi71d
.3flbo'A rt S at'AG= ~L~~
,,nktxt=c?ned her;:#::n, are incorporated by refcrence ati t~ic;.r i.=nbr~ty:
i:a case of conflict, ;I:e
p;. -zent 5pC c3 fic`e;.`iYoEa. including df,.~.:'t.3.~tions,,, will control.
i1o~~ii?g
Other f~y Ftlfeatures a~i'~~:~il~~~in~ii~g<:S of the A1 inv4'=tition i51a7 1
be _:om the : iCy;
t:vt<a1E:~.~d. ~~~ senptit?n and fi4.E.1reZa, wid from ~iai: claims.
25 .~~~SCR.1.PFIO.~ OF MtANVING~
~ !G. .EA is a I:Iic 4:?-re`2 IlluJirit.l1g the t3h-aiiwo lil
flu{.}ri..si.#;;ri:. K:ftcn-sItv w.~t'ri
inc.e.a~ing conunAraÃiors .?f:-neÃhy;ene blue (MB) Ati pha~spla te buk?cxe:d
,.x..nz:a<
~-:lied squarc::), 100% f~Ãtal bs}-vine serun ÃFBS, opez-l. triangles pointing
rt=p. .iashed :n,,;r or
,
.ini ONR~~x As open triangles pi?inti.ng down, solid llnt: f.
30 FIGs. I B and I C <3.rk- bar ~;~r.~p~~s ill~.s,vt=~.~hn~. ~ak abso=.~3~awwo
~~avQ c,.g:}: Ã z B?
PG;ak flE:o.LSi:ui:ca; ein3S;ilon wc3-ve;.41Igth (: C) for II1G:~eas1ng
c<3.i"#.Genttcltxous ox MA.~s iil AI~~~
~ ~lack ihars'. Ft3~ (g-roy biars), or MtOH (wh;:te bars).
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
~ i~ r~~) )tYty~:~ F'~" iF ]p}Sy+)t'iJpr;.~ise~.~iA~~.~\. e}.i. ynl+~~ t ~w-
eZ.rwp.`-~'~.~4 ZSL. t¾- ,~<'p l.~=~LB }
~ ~n :..~' is a ~l t3~i w~2~3~ C~~ ~ .{.S.i.~[Ya~~'t~A \. i~i.
':[~\
pl'I --- 6. M3c'li:'~c conditions, el.ilia witl1 ttit: addition i,1f Sodium
Ase, c2s:.&bic,
i3u`xd..
FIG. 21 k ah~l of n31:o images of a rat heart c.:t sequential ,'dme "Noinis
(03 zt?i:?
i] au~v.1 12 seconds (~i-iai.3df$:.= rt-.~w)T <~titb -5 miluies
(bo4t1)ir,.'ow)) after 1~ .adul.~nisLraF.iVn: 01`:i:B,
...
T ! ~~4,
t- ~r t3nz-.. x{h aue, s~'. ~FSwn in the a 1~:'l~: s . i:olunnn, near ~ (N.
Ix\,7 'Sy .l;a?Sj
:~ c3 w a~i."s4'1...xTz ~:>
~..~31,.
4e (merged) w'e ~~~i?i~'l'a tI1 t:7 , xg~fti i. ?iei I1n..
s.E3ddla' and aA3Vio1'.=NIR (3vefl.? i:ilid,
4? c i ati a s:iiit`7le ai?iage that displays both a:lafom~' (color video)
imCA function (IR:
flu'qui:JCelale), e110 M fli?ore5C.w:inct; image was peeuii#3-cE,li?s'ed in
IT.i">ii. +rt:e: wli; overlaid
Wi.ai field.
i:.~a 100"r i 3 i3i'a pilrenY y ;3n top of ihi` color vi d4=o image of the
Si?inC sur g
FrG, .3A is a p,.vir of 1~iib graphs ali-ust4-atizig sig:sa.to baQkgr13u-nd
ratio Ã;~~I() in
fonrrnom bile d#,li:i (CBD) versus pa-Ili:~rea4 (left graph) or liver (right
giaph)s -at sei.]u+'.ilt`Ll
:`Ã:E~~'~e points a-l'ter of MB.
e'IG. 3B ts <311" i# of tii?lago` of rat CBD 1171agef: al`Ee1'
ivad:71E,11stF'<7.Ã;.t1i: of N14.13.
f Color i n?c~3 ( ~ ~ ~ shown in ~~L ~G~Ii?C3lLÃ:73, i ~?<3:I' 1n3~i i'S t~
(N ~`~ ~ A71.T:t:;s i~7~i, sa;.-$3 r',~.E3 ~.1~. the
inidiil3 and color/N Lk overlay .'1'lic:l~'.,.~~in (m{:3'ged) a3'C.~ shown
in thc'`. I?'O, it s'rU:iln:x .
1, 1G. 4d~,:` a trio ol iL1dag'k. =s of rat C:Z?~~ imaggeZJ after U:i_'=,5
aduw4i.ii.;4.i~.C1o~n of iYb~~:
Color innge:o- i3..e shown in thL' ~~t-I column, near 1niT`uk'Oi~ (N1k)
urkars~ s-, anresh3wA, in tiv
7?Eddit cLaf iinis and c3lt?r%NUR overlay lmagc4 (:]`1En4?4) i3ri=.
shuw.E#.IÃ. lllu F:gIit :=i?`uI1~.;n.
2 0, FiG. 5 is a set of 'ti}L; images ltfCBD imaging ~.'itf;, L~~ ~ (top
row), as\^.~C`) "TI:.ii~.~.~ai:;
~
row), aÃ`:tt CWuO0Cr'1 Co4SEao33 row), wh'fcll are able. to pii7vId.'
:T~iLii:ii`r?"u tl+?f. {}f the bFl%~-
dÃ.cÃ. 10 :,3tg: zafl..:5 i~~~~~.~~~. ~~r ~~ ~:~ . of 1000 ~Ã:~~~ It_.~~-
.~~?~~ :~f? liI:_ of 100 .,~~ ~:E~~: :3~~.(~t x two
NIR flux3-rt3v23horU si.3aLtt2iZf''x nso po'=toi voin of the rat<3. Shi?wi-
F i1T'S: a still fri}n col.>8 i:l{:e,.=t
(RAT.). .~~R fl3:#.Z}esi;:.imi:`s'.~~ (3n3.d3.~lf:=)> c.$.I3.d mer{~~d,
images of the two Sr4_^".l.tf. CBD i:i.t.ii bi::.
~'~'~~ ~' ,~? Lw~s' no dye stacking :?
2>; 4'. ;.Ew'.iizei by a.13 ...~ ci"S.1`?.rii`3t agents (`srri3vl,hcadS),
bLit C~ :~~_ ~.la~.
~
thi. Rv<:r and ai~ijhz:s- SBR thaii Ã.he sst~iez two. N-UR Ia?~iocescent
ex,:3og.aa=~.~ time as'200in,,se::.,
~~ [G5, :;.A-D at`e line ~,~3'il~ih4 ~llu5$rtltll"ig quantification of fl;Z
ti 4a E ~~ ~ i7711xS`iC3}"
and signal :o baz.k~r.ou:~~~~ raW (SBR) kinetics for CW8,300CA. =Th;: S.13M
(rzwa:r., :1- Sxt:y=' of
thi. Lom3-ni5`e`1 i:lS; duct a`i:li1:i'4'f,'~ to the p:~.!?uvt`.5 (6A)
u$~;ad liver (Ã1B) wwI'f} qI.1.<ai{.dficd o-vo<y
..,: finw lrjicÃ~iork of'50gl.: of l.f?, 20 50, ar:i~ 100 ~;.-,M ( v~ 800CA
solution. fl.+,? SFiR tt) fl%i: Pi:l.Ili:.:G`s4 (6C) w.tid livcr (61)) wL-
`rC: al5o i:aE..fab::eF. cafle pt?.Ii'cd3
i:j,., t&.~n of 50 ~LL of 10. 20, 50, and 1 00 }iM CWSOOCA solution.
6
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
~I~:5 ~..'i. t ~ '~13 are sets c?f'fil~.~~cÃ~ Ã~~~~:3r>~:s of ~.'~3I~ ~~~~:p-
;~- ~.~~.~ .C..~iÃ~~'~~r~v~:'B`~~~~.:~
~, 7A :~t a:Za r f. images a
;di~mfii:cf~ under nonnt3~ ~~~iditioiis (i:~) mc3. after the in~ertion of 3
bead~ of 2>5mi:n and ;
bt ti: of'.5mm Ind3ainc.tcx into the C`BD f-rom p3pi11:avaÃ.er Ã;713;. '~ n-
A. ofia50PM
CW800CA (7,5 i.fg / kg total fllloiopht).:e) w#s 1njeczC'd Fn:Ã'%ivuil?a.is1
4 and #.i;i7gt:=d t3. ` ~
.`..` m#.si:~ Shown are s:1llu from cil?orb'adeo #:eaÃ~~ NTIR
flZ:#t?I'4,s',`enirG'. (i:'.lit fize), and z:3fu.a;ryf'd
i2I...3ga:;i of the two L`. fs.3`x0. Allbcldti
could bc: t t,.tccteN ~.3t:r-E.,ctxy M each tr#Ct1.
FIG. 7C is 21t:i.; of a)'$aga',:> of pig ~LID .']Ft.IL'.ea aat:r
e1G,aii_i.z3:i4Ãrate<`,E# of M13 rkil:d
ICG. .e..~`~ ll. color anagey'#niddli-:<. W X3 N I.R. fl#.iC.3-r"'SGE.'.i1Ã 1-
maggo (~.~~0i; #~7#~',, and ~ghÃ,.
I ~~ _~~~efl,Mene blu~., NIRF ~~~~~~~ (MO rm). In th:>s4 :mages. ICG
~~ghltt;~.~tsi blood vcsEo.s.,
wh#ie .%,l33 shows the CBDa
F K" 8 : a [ i'.t's of i 3:0Ã' Imagi 4 (?~ tht I`a46 1L duct An's? ~3~,3i11'uw
in a pig. Shol`,,a zir:'
sfills. famn, color vÃdeo, (left), NIR fluorescence (nuc3d:Ie), ui-i3 -
rner=,ec~ i~.na,.4es of ~ V;;o
("Ili_a h t) .
1:: 9.'IG.,7' "J a set of twvlTb': fi-nag'`.s of thoracic l~vta.L
#.na~..~~.~i#ig %3 t,: rat i-#siÃg ICGE (liY:(-s
coaunii~). I~' GH:3A. (second ccfluann), C~'~ SOOCA (third cohm-ir ), and
I3SA800
:.c3aumm), Shown are stills ~~om color video ktop row), NIR. _i" uorescencv
f~-x)zÃo,.m, ro,~m)>
-i.#id mkmc`iCl i#anarc?'iasoa` fhe vwJ(m#.ddlL~~o-w).
~ 10>, a0aÃ3d 11 i3re rll2e '~,-~mpi3s illustrating qi1;:1n~E1ficaÃ.if?n ?a
Ã,.7E fl..3oi'i:sc:=#.:E:o
2C eTni"sion and ~ipal to backgmu#>d ratio (SBR) a~ine..iis f:or CWS,;>5OCA:
11w SBR (#~~.ealn
SE .''aA) the ureter relative if_) Ãh'':`. k.kine'1+` (10) and
abC.t3a1"I3$3.:~~~_~ wall (11) 3'Yere q~i:mir.tC;i:
over 3m.-ras~enous injection of 50 p1; of 10, 20a 50, and 100 z'Nt CWHi:#i_;A
Sz?atEi5n,
1, aCf. s 14, is a scrics of t1~~~ec images of tit.,ew T-napping,n tA r':aÃ.
Sb:i5wm are qil:y
24 video ~~eff,'l3õ NIR ykoke5e#.'.i3ce i~in1ddle~1 and illf''~%;C.S.i 1'm#g~
of the two t~"I4;=zft
F:f_:s< 13 at3.d. 14 i'-1:#'c ge 11ph;i illi.15traÃlli". th'~,.,`' 1#::
a`i.ilt`'s of IAIPIYCh`ni3ss >'sP+,.c~Ãi'olllt'~~:r3,
meia,bo3aY i:.,Tatvsis of CW8\i'Li carboxylic a4.t.dIAfte3 excretion .L-itti?
u2iA4e. ?lssbiiwn, by th'.-~
:~i.,o ey:?Lnce cÃ.ro#l#uÃz3gt'i.#pi], the dye i`tppt;:aI3 as essentially ii
single pcak w~th, the ?E; FIG. 15 iS 14=2'#.e;i t;f o#gj1Ã C:SifJi `G'li~eo,
(left i%oluTI1F7; and neaa I#.,,&ar:.ÃF flEao..L_4u: -riC'e
(i'#;;~i: cLrla#an) Ã,..~t~:g,',s of ~.`s~ti.la during intravenous
IIIr'cÃ'.iC?a3. 3. 0.5 m;j. (14 I#g.I;Y;
:a:~?~n141e ,~`,.'''.11 (1CG) GIlÃ~) 3 .~'7S kg ~rE^~~'~,5h1Te pig. Shown
Fe~#'L'= pra -iII;G'ction
i3.e.tofi43#.3rG:?c#c..` ~'i5p), [1:rà eAi.#E fiffliag :it 5 ~oo p'a~t-
Y#ijvctC3n ' bC;c?nd row) and venous
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
filling at 10 se~:. post-irkõ~.~:ctÃon. Note tliag the nipple in thefie,~~
(top right) .Menr~~~~--, as aTF
additional internal ct:}.,1t3C?: -IfFi F?stQfiai v:s'.venCtiI:Is phasws.
~.. ~ and r + ~..~Y1
~`Ii.~. 16 :s a '~i}z3..a~:<l [)
f s..:~Eei%4 Tr7:l~Fgi'3 of Fl~~:.k ~l~~li.lf~= blue ?.. t:~.C_t~3 IV la?j:.
..a~i;I?s
~~~o~~=i.ig C131> a:s? CystiÃ:Atte.Ãy (`vA), as ~.Ye1l as coronary arteFy, in
an intact (top 3
rows) and : e;:cwYte~ (boÃ.o,::~ row) lioart.
FR:5: 17 is a paFts;l of Amkges sliomdng the IesuFss og parat3$4`I'S?.iid
gFaTFc. .i]uIging.
DETAILEW~~~SCRl ~.~'T:l0N
i,i:traopi :ati?<'~ im<7~,~"Vtt3g is eU.indispensable dTi3g,,?stFc, tool for
ida''ntFficat1.Ã3I1 of
noIF?'3ti:i or ~~~ eÃ~i..T?~33 anaio:nF.i.i:l~ AT`n3.ct[F-Ie5. Si3 gt C?:3s
usually F1~o flLFo: ? t Zfpt%, kt,FSo:!;?(.
i u ~~~jec}-ion oflhÃg& doses of a blue dye (e:gr, iiiUtli}{leiio blue (MBf,
L.3rrnk~h,'.:zz.rid.1~1, rir
ivan's B1~~~~, or radÃÃ:tschnfigrvFphy for intrF~opc.r.Ftive imeigiag. I'h4so
m0 th,3~~s QtF.r~..~vl
,play;igy~?fiCif.nt >s3ics, however, F:acii methC)d l;.ay stJi:ii`
~raLvbs..i.k~, sF:ii,...3..z:ti m..,:adF.;7ti0114
3"`a:iSC?viati,d wFtl1 .``+iuC:s?'GFfiC:i;..?py mId
1a:Fd1(?Sl:iIltl`'s~.ryl~.`~hy), low SpC',+.i"CÃ``..}% a.1L, lo:T.:
sei
-.s.fi.v:iv (y;,Tual biue, dye:j), lrt soai-te cases, tirr exasn>~le IMB, the.
<Ia.sos r~.:.~u::e~.~. tf)
aa.z#Fc-ve vzsuei~:l- a~~3c:11tÃtzti,atiz:371 m-t' associated w1d1 significant
t{3x1c1ty.
t>?C: mQiChods described hemin ts,3k4 iilivantiage of tl14 ~Y opeF`t.FF,'..4'
oa. fluorescent dyes,
e;g., invisible light (IL) aha(92"esct:I3t dyes, to provide &afE,4
fipa:'.i,F~; .e, a;?d Sia..`.s.fF.vS:;= raetE'f~fds-
ior l<ib';-'li.Fx4. and d5:tocthng anatomical Tht.Zo;-..ns:t3:tt'3d.4`s
11W:Wwo
thwr LFSQ of d v:: 's tha~ Fa~ are '~vltcrod aiid excreted by tr ~~. ~`
kiainey (¾c?F- ,~~~.~, ` ~l`rFg of L' L'~C. xh)i?: 120 taken up by the 3:ver
md. seL~retedin,..} bele (i`o.F= latieIffig of-th~.~ biliary .'eo). `Fho
pFcfrru.;
d~c. <b.+.4=aot si.gn'aficCiniaYV 3''Zii:Fb.]SJibod or m'rvt[2btilizf,.d ~.CS
the lqoE4y .Thf: m.od.\?d,,, t, - LS,:':
.. . .. . . . . . . ~p.+.:.:a
?3F diA'#.Fa~ a kni1LY7? dye tC3 affect its filtration and n?zt:3:?t1i:l
prL)s14~I':I~..4 ~i'r exe3.mple,
s
~'~t~T~1',:~:~~C?#.i f~3'~. a ia~{.i:;~ f;:~.3.~a bi: uh~ .?~. to
.~i~.~.~3.~i~~~.Cii;1`~tr~ z~'{f~.FeC:~ liver uptake, t.=?y
~la";.F.T~.`1i~~~r
c{y:" iiito~ the urintFrv tract.
'Mc..th;jd;
".~ie 3method5 desoibFr:d lFerc;iii can bc, practiced wÃf~i any intr;Foperat.;
e :~Ma
19i
ywte.i'F.l fl)z3E Caa dduct 8z3vÃsÃi?.(t:llga}1i. (IL) flE3t?r~scenC'.e i'yt
vivo, C.g., ihL. 'Systo.+`~F1s described
in Dc>= Gtmid :i~~d FraÃz&ioÃd, Toelmol. Cancer Res. Treat. 2(6).553-.62
(2003), U.S. P<u
App. Pub. ~:` ~ ~~t~'~i~~Zila ~~ i3 ? ~ f?, ~ ` i~.jS~). lp1.3.~. ~eÃ?.
~,?~~õ~ ",~?~"~~~3~ :`.~:?
~ ). tz ~~-r~.~~ i~~F~a~ ~. aF,~ U.S. ~Pat.
;:axangi~~ ~, U.S. . ~ , ~i3~~. Pub. Now ?:~05:/t~t~'0~~L i ;r umioF,. et : i
, mFd U3. P ;. Ap;.
Pub. 'klon ; 00:?:`ts3:''s2?2l W,FT'angii?Fli, -all of which are ln'4'~o-
,ps3raxted hC,'.i cÃI: byHi.fLT'cI':;~:;~=.
~
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
T}.> ~~-keth-ot:s described herein Lam be used : .,; pLirt of an ~TTaaRrF;.~~
S-ytsÃem" . g,
-ph1Ãn or tE?J.w:og
t^F3h?.c 2Ã27kgIng sysioÃa1> ioà high sensitivity da,"tCs%tt{.3Ã3 of flEaoI'E
thus, :llc .,>Zwi:>ztds are ideal ,t3i intraoperative in1zx~4jiraf. Mo;t c~
vc;', the n1e`h.3i,4 described
hek'E'.Yi.n L'ian be \a.E...+.d f+ pt`E'3Y%ide ci5lo1', fluC?-roscvnce. and
r.E7eEMf:fx images y ~~ keep ~ # ~ ? @' ~.{ ' YQ\ real ~'S
:~`~lfif~: :3xL;ZG'~ i~~~~4 ?`t~~ to ~v~}3 ~-~!= i of f ,3 ~ ~ , ;~f?~E
~L:vx1~~T C3~'~=a ~#~L S~;z~CS~ z~i~~~ IÃ .~..,na
iah 1Fr,f~-aT~: c:~. procedures i~3'.'=~' C:s3`~44-.',S3i.lI~r~,-3.,.
~~k.'~CI]'l~l`Ã1g on t-~:f'. RtI':.'~3:~ih of the flL`at?.c?4FvC:nS.t., and
f11c %E3SõaEaEin, ;aaid size of jthE? `it.'13`Nt`iir`.'. desired :C) be #:TIa-
gL.'.d, zaE:t3rosoi.x1i:E:= thatt~'r Isp W
C~- the L>ai >2~iI~.`i from +x}Ã. sttÃfc?t E? cvli1 be E;LaaE,~t'if 'iYÃt-h
pi.<aIi<3a : elle Z .s?Ã3cQ 3ÃÃl:xVg[
Deeper fisstxs .,aIi bL III:ag;..d .14:i1.~ t~sÃiTtiSt?t<1~:.~7;~a
2371~d#,'li'f.~ methods, such .E ~.~'~.i~ #.tL:~~~'-
adEt?;.iax33 ;bott?,s.] in"Ta':1.di??n. E?; s.Imc<,dÃ3I11a:17i t&h$1~qv::i,
wA11c?3 will likely ~.::~tc31el (t`<.`.a1tIl
dc,~7 ~ ~~ , r ~r #~~ ~
4ti..a;`'~.~1'~)Ã~ to a.,.~ rc~, i'~; 4- tC~ I i O~iwi,a ~~%~Ã?g~
(a'i'.~'ie%Z'4`+:td i~~. r~',l,;tci t:t.'~:'6i~'aL:i: ct i3.~.~. b~~i.
`l~?A'~i. ~..F.at~~~~A.
B1:5 , 6z641-3350 02) .ia3i~ Nb,.i3f r3ri.sl3s ea aL, EtIl`. R<3.cI`aO..,
13:195208 (2004).
~.~xv x:lg~:x~.if~la sf,.~lci=ted aiia3E?m9.cai: structures, a`k'`+'fi main
broad atÃ,-0:.u~ic~, can be 1:S00,
ACia' i~1tIi i ei trt}1> of ~~ie dyes Lx~:S~:~`i~r'v'E~ ~31~'I'4`,31~.
yc pass throtigat the a4guan
iu 1) hk#raLenotÃs ;Ã~Jccfion. 11-ii.s.nnelhoÃ:f roqu:itesffaa: the d,
P by the liver or #~id.''sety
2Ã:t4~thC; bod~~yfl-uiEq e :_f.ycirl,.=ul`s.:'tL in tFic bAood.~`stroarn,
b''c.:tak1:n -ti
and sZ`"4.ra4d .3vW bile or 's3riÃi'L, aFld2
2) Di~eit i,.~js>cfitrn :1l1tE3 the S$rui:tu2'E' to be 4'i: ualÃzr>~. Diref
#a.cÃ.1on also has twr',o
j`:>,;~~~~'.1i}IeS;
20 i An-tt.'wgr:ade inJG:i;'Li0}F'a, e.#,~.~'.. ÃÃ]~E~~1s3I:F w: the "tG~i`y
of t`~4 s~`F:-~S:`¾ti~Ã'~., b'ciY.o.C..a..
1-lro, fjE>c fol1.owi th~.~ normal iflow of the bcfdi~~~ BLi:Ãd.; o g-, z3fle:-
iÃ:iec;ion into à he lell
o; r`gf-: h ep.~~ic duq, the dyÃ: floY,~ s distally to ~,is,Ãaai.re C'BD, and
:~) Retrograde i~~ec:ion, e.g., caÃÃzaulation of the oz'ifioe of :; s4ru:
RIx`Q,
which involves fqjeist3ng the dZ',-. against the naaural st2've`.liTt of the
rlt.'~id.
f ~
j1?"'.~ urea0rs i:"E3sanE:c~ the kicitlexa' to the bladdf't: CU,'t1;11.1g un-
ne hadun with toxins
absorbed tom t1he blt363dti.rL'=ifln by the ktdney~S, leakage {J- which I.ntc?
:li''r`< ~.~o~..y :Mty +:an
i.?amse peÃiÃE?n:tfi;. During aibdom:Enal surgt;=ri'.s stiE:li as caosarian
hcc,oris :.Etad ut':klt3;u-:teA
t:r s~ix~ >:~,` ~r Csal .~~i~~i. ~:1c S11i'gt.+:?Ii be iiEblE.; to identify
t1he ureters, e.;.-',., to ei!''i;it;.3
damaging 3hi."':rn, or W iF'pai-F them after aatr<JME;1`~ic or ~xtc'>mal dsla-
na,,t,v;E;.,
9
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
Metb,rd: ym ama~.ing the uÃ'etefs 'iÃfcl:Ãde if~jed:yÃg wn irfvzs'Ne lig}4t
flu"o;:opho:`e
fõ #LF) into th,~'Y+i.oi.3dsU.eaÃ>.i, Y.ir CLit~.ect. cannulation, C+ithvr
':.a:[>tvsrot~..Y,rad4 oi. LeLf o.`'ty2:a.dc i} fLi Lht:
ureters 3F bliFS.iili .=?. ~,~'l.Ich that it appears :TF the F:fF":f3e
'~~TS.:i$.n.
fmagt; tbe 3,F[etvF's= F: >s desi1"-a~.~lo :'F? E,ICo an 11- fl3.Fi:F1`t.St.-
C;FAt c:.+mp+:,tild that is
~+ /; by ~y q }¾ ,l ,c ty ty [} y~
.~il~~~+F'J..~4'if~Y~ f33G~,~2F.aaiii=@=~ A.il.f.? t~3~. :#~"1#:t:. S~J
\4.~4S/<y5k Maa'.~.~1.rlAti2 ~'C~tlt ~f ~'[.+t-}.a Z' i .`A~ ;'\ [.YJ \jf~
~ have iA t.: t~2 ~a o11c4F~:_a
:a:FC.F2)e?:e.1 of sGSs, than 5 Ilfnz; are hydf'oj~hiht;; and are
TlotsTgF7.FfiCr1.F"7tly positively :.lcxi .ged.
Agents that c:}ef: i,+: used iÃ1?:.lf3.dL'NIR flu't?rC7p.lf}i=e`~' S?ic:1 as
Fa3'vlh?`lÃ:'ile i3ba,. n;y:Y,86,
CWSOO-('AQ Cu5:5; Cy"¾ C3 i.S. :(Rdz=4:`Ã'-,A~OOCW (LICOR), and IRAYb'r 8
(:_.ICOR:).
T;.ac lci`i:l 0, i hydfo :3a3i Ic1ty of a %f):11.poaFf'nd pt a5'S a role ftl
d3FC.ci.ing #.:":1E.alkc 'ÃLk :hw
kitiFii:y alaaVor liver; tho>bfo5e, t.3eh5rt..ie3pllfla:dty of -a
mE;dF.fiak3lF;=: ctgÃ,ait ca.~fy a3e zenS...FF. asw'\~~-n
s''.g.T by 3n<:=Ã'e<`F.sIng the aQ+'G'I t'sf SulphC5z123:t3f.}Ila to increase
uptake :av~=':" f.ind. a3Ã" : idlivy. .1
fsL=..ti.'t:>Fbed h'L'r5:`=ixl, c3:,:TLMs 1',:;f3:t t3re u'z3sul}:ah1'tnatGd.
ITe.?i.iCyst1lph(;}:1a-tc:i,Y or fl1si.F_pl-iotlzl:.i:d art.
g=,n<,rYiÃly fap3d`yf uequz.stc~od b-y thc lõJcõr. btit a.Fr:. nc}t. .creE;.d
Ã~~to bilu officio.3fly, .Ã.d rt;
thus not partacWhFrl}? useful iil' xnI.F]9u t13e b$h<f.T'y lF t',t<, Ageizi:w
that ;.F3'e ti7'#Sf.FaP}eO;3ZF~.S`,(l,
mulplf:mF3..'4d, Per-aa3.suEphiDne:;d, or hepi.mulphon,':3.tt`,d are mt7w''
likely to be secf'E''tLi3 bi
thà lxvxE?" fTit[3 bile, .~.f~d aF'e also x7.?oTG likely >~) be available in
the circulation s#.~.. :f?~ S3tat~z;
by the kidney into and aF''L tt311: usa:ff.ll fci1` i21,f1g'ÃI'Ãg the
u#'ct+'=f':^ a.2Cl hFl, i}'y :~oc,
Agents that can ~,"`',i~, \ ~ ;-nodafi~.f~ h F~ this rfF1~,l:1~~t include
F=~'u~i, ! J 5 i~'~~E:.:~ such 4..`~t'5 (_~ ~;rtF~..;~~.c. ~a =~\
~ ~'i'3i. P1~:~. < ;J ;)...m
BiL.3, ca1.'.nf..vs); c g. Lv, su:&ofa.ati(..}n.
'Ont:a a-f'v n[$tF.3rfiil1' taken 3.Fp by she liver wllI'.'ià injected
sytitC,mFLd:l y }S~
2 0 SoITif wg
:5efec.t.Ã'4'5:l4 liii?s.I the u1'ti irSj such agents should be modified,
e.g., by <f.d:f.a.f,3.F t,: `sF i7F ?~f f '
s:;ch~ as~ .F PEG i.;,, by p~gy1at3i;n, that p:i'uvents t.fptak:: by ti1u
A7ve.x,`to :fnp;o..='::' their
Ske ir,Ff itv zbr the ureters. Methods for pegylsitif:g ki}Fx1poF.:ta~dti
<.i.F`e knowi1 i?a ::h art.
~
25 ;.~di is~~;j' ~z ~:.' d':?'t ,'~,:Fx'$r,.
lF1~,.F'a't?pt,.rz3tFvi:, :holaÃIgogi.`e3'phy (10C) li.~-s beiia Shf3wf1
tf.~? dcC,l;:~se the risk of
ct;fs.>11or., lb1le dFEWt injury during 3:m"asbvE' S[#.YgFi;;~l procedures
such as L:holi.cysiWtfFfey.
IFlv'lsHe LFgh~ (~~~) ftut3F"esF:F<:13LL`=b.lae duct imaging offers so4'i,F'al
wdv:F.F~tages over siKtl,da?'i
IOF with x-rays including no ioF1Iz1Tigk'i3fllEFtif?f3., Ie't:Fl-iImS.'
fa"flc3wTf.g. and sf"aaf..fii.a:'i`4Q:l.i:i
31 Ã; :rnagpFng et:tia ÃtÃ~:= c,.~loi- vFdoo fimige of the surgical fxe:d.
In tl?c. moahctda described here.in. 11_.FsÃhat are. fil :oreai by the liver
into d_Ãe .?:iae
stya:<3li1 i3lLo3y V3HiFi.a:%3L<7.t3on of structures of tt1e biliary sF s,.
.('S including tlic c:;F..3FI`:o.'^: bf lc : uC i
~~~
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
(4.=BD), w.lÃcr? is vulnerable to a4'Cld`s'.mal inj~uS` (bui`iI'Ãg Si.irgei'i
as it is hidden by, c1i-ld
f.f.)l-n, otlii<,r fis:?u'L.S.
Methods fi~a firmging the biliahy tree includt. intecting, an ILF into tiw
bkxodst;vwa:
Si:idh ~'ii-lat it i3:pp~am in the bile 5:7`ei3.ETF. In 4eT:eY'i>f,
k::.iE?si^. 4:go.ilts ~l-i`ai$ w.Ãl""s label Li14 lIÃi'>i:OÃ'S
; w.ll,als:? 1abel t~e bilian t:ee themtb:e; agcaÃs t~~~~t have pZopezti : ~
~~h11:lar to ÃI ho,e
d;.=s...>zb~.~ above for labeling ureters can be used. ILFs th~.Ã ewn be used
inc:ude NIR
fl[l.fYi`i>a h{3m,~'_.+ such bi5 31i.ctl4ylunc blue, il7d'l)cyfiTl].nS',
ta~?,I'Ã?t'.:il (J~.~G)$ IR-7~~.S6,, and CZ`:+ SOO-CA>
z~y,5. ?, Cy:, iYs 7.5, mi~ IRDys;''-,~", 8,
F.51' rapid :inds.,}eL:ive Imi;3?>ng of the bilr:l#'y tice, a;-i IL4 can be
iÃ``,...ca,'d i$,'ei:.ÃIS`
iii,t{:f the portal win. Direct c<#.17i.11i:3l7C>iI in1.i'~ `.l1e i'igl1t or
li:ft .C.,pa.l:i:= duct for anterograde fiT
i'i'broÃ71'e'Ã.ixt; labeling of the b1.3iw-y`iiot.'. Incl:adlilg the CBD
C=:.'+,~. al st1 bc LasLd,
:.hor<.cÃ~ diict is an impz~~-tant partoi~ the lynnj~aatac s; ;;ten'i, ce~lice-
in:. most o'a-
Ã:1 ti~.~., ~.~~"Ã:~~~.~'3. in i`.~li; ~.~L3~4' i?Ã..'?i3 t~~';S3I]:1I3~; it
i31Ã.#3 f`l.a:: Jy4Ã(;71~~11?. Cia'E:=L2IatiC3i1. i h,.:t:>, >zzcthL?CS`u` fo:
1~`t'lcz.g,ia"ig the thf}:iloic d...d include IIIji:=i:t.ÃIi,~d.~,, e.`~.i7 IL
fluorescent compound, < if., a
E.f.?Ã'~.'SGe,.t ct9aut:1dy into a lymph n4_>dc, e..g:. the inguinal iE.
lylpht~CSLxS;, and dowE.,t.;:i3.if i;
fl
fl:aofeJcS ei.: sEpmal &om tlio dwIa.GÃ.c du4=t,
lÃa some, eT11bE3dit::~enÃ.J, flti'= i31effia}ds i?.t;ad:.l'.Ãbt:=d hÃ:.'eiIi
PoÃ" i.ma,~,'~n,? :tic ;hoi"acic duct
t
l:icliiixe tffie 3:#i:i:,' of 1~~~~th'S' C:lla`.: _hw as the ILE
"`;m 3,3~i?: rai~
'Th;. a$3314;4` to cl`~`.'~ai'iv i.dt:ilfiiv L;`is+;.'.ulatul"c in Ã'i;ia-
wmi,' during surgical õD*'f3r:t. .dum;
y Wc3lild : ., ..ÃiL'i~;sllii?bl: fii.s-? :a3G
sL~cli as 4z1rSziac surgery, neldiLx5urg<1y, and wTf'3iei'ill siS#g'__'
~ s:~ -~w..}Ã~. As described lac>.~-ci~~., it has 1~e~:t~ z~i:~~:o~~c;,r~:~~
:liat ILFs, ~..~-i~::~.c~.i i~;.~-~s~.=t~~vI..ne ~.~luw
Li~i b4 usi:ti foi "Ztaa ti3'i3.s~ li-t tÃr:gx41gr:tpaty of F)lood .'k:
sel,,', t o al Ir?'w iihs LS;,t i.Ã fx
vcs'-,~`,Cl ps` tt`Ã::i.y; 'wiwh a gt'?eid sÃgni3.l-w-backg2Ã3u;.7d ratio.
.i xx4F;..~,i 'tI~JS Ir{)t` d{!.~'f:`s:Ã3t'2tF!^t#~'sri3'i.':f'gtxtr~r
3u> :n g,::.s ii~i. ~'~;t~f~~aSÃr~lititie ~irgir4' is ~?>i~ l un
~lz'itiC~1~3.~~ of :Ã6S~:u,31..`=~~~:i11~:~`it and
tissue most ComplLx i-econ;t-riiGiivc ~,woc:edure: make use of`flwps,"
i!'hici7. are blocks of tissue that can be rT:1ob'd C]3'
ii"'t1319pl<:i:i7ts(:=d based o$r, t3#Q an: wm:2:Ã'
chaI,.3ctL=r1s11a=s. of the blood supply to ~ti>;: 3.~a;?; Si.tch fl:3pii may
be (,3:'E-ipt3St'd of skin,
I I
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
muscle, e}43?.1e, rr.,`.dt?F;1, n`~.avep b:i,.)o5"f vessel, $õ>i-('.st1Ã"1:a,
:'1i.1d vitÃi(x`L}.s combinations of Ãlw6c
3onaiiClni h.k:;
ÃypC;`s of ÃA5s1ic. These xiapS ceO.tl be moved from a "donor" site, whos1
S.#",I.i
ac-equ~~le mo:;?td?tye Ão the "reCÃp?crt" site, ~.vhkcre rÃ:.constnioÃior,
?;s, neÃ:=d,,A For uAan,xA?,e;
a l:om`',a_3{3s3to 1': `S. : flap of ~ibua r3~id overlying skin ~.`,aii b
i:se{"~ to recon: tmC.t a naandiblo
airdflooE"-:)f-3nt?uth defect fol~oit3i11g a lÃ54jor c+Ã"a. ~=~~ri~=t":.
T:.SoitiC3i'Ã. Ota?:;i' ex<3;?,:?ia'::
include IleÃa;ts~'-nl"F'wiahz fli3p --, tl?I'f%tG:lal Tlt'rve PaTalV=kda5
fibi1lF3., o5tof_lC,'ra3.fwS3us .;.2tp:~ fiuÃ'
mandible a3~d oral reci9t:sInuCtit`.+Ã2, bAm:st 2'.`L<3tlstI'Ã].'L'Ã?o11 fol>o-
wln'k< b,ei4t caslcex, lower
z. ;alxemity salvage following tra~uri<:, airÃd total penile
rof:..t?nstruction,
The s"ing.Ãw: most lth pi`,T'ta3:tt} dt::F:a .kFigv i3T1d t,lT.1:1q".aliL'
s3gI_a.t,I:Ã~~ C}fx flap Ã~ li:~
~~:,.. ...~s~~" ~~ .~~~?~'i~?~ts:~~'. if Ã~~Ã~Vascular s4Ippl}'t~~ a fla~? is
not t~~:t~~Ã: at~., Ãhe[lap =.~~;;
lxwo:ia:e and fa?3 whtin t:'atlsfQrrGd to its new PosItÃom 11-1c s= aJ::ula;'
S'~,:Ãpp.,, to a
~ 1~. ~ ,` ; .u.õ.-~it~sà be adequate ~ supply > ! ._~-Ã~: ;~l:a::l.~~,...~: t
a~~ t ~.
~?~x?~.:~ of x?~~,zc.=, ~~~3s~{,~ as a `~s.=c~:~~ÃÃf? ~:a~?~p. a ,se
>:otiie.;`ti"'~m11~~_ be eic~~'f.E;:F. .:3.c3znta1d3<:d during thr.', surgery
(pedYcl'vd _q:1p)., or if -i I'...i:yt bÃ3
iiL c :~.t: ~ ~ ~ief a',1 } ~ r~" o2 the body ' to I c.1cl`:~, y}i~. st `~.=,
I; ;-; `:
-~~~c~ ~:: ~~. à `~.~..tà ~;i.:Ãz~~, r -~~.~ ~~E
Ã"'K:.al3a5t33nto5ot:` to blood vessels in Ãt'1: ie=L.":1pLfu13.t z3.1'i:,il_
LÃs-iÃ;'g microsurgical cal Ãef h211q.3.t" c3s3d
rnicrZ?::tLat gici:l inst-rumCnt:e.Ãiti'~~~ (free fli3p)~
I F3C ,zw.rrphi:=f Ftl blood ~ sv,,pplk to the slti iÃ3 1F1 i.n35: fi c4~
~lap donor sites EÃ';aes ft,.t1.;,
Y
one of t'i ? 4 ~35 S~`S.. ]SZt. 4S~tt.. "( 1~tF"4,:i.=te ~..3I..;i~.\ A tl`k=
~Y g: j'Y
Si~:: S,tii~::C.=`=,='. :~~-'~3~Ãf~C',lEÃ:~~ ~.'t~,: which course
~s~iV'~33:1~3~,
A9eYfcli"i3tor vessels, WhÃL=h pÃ:',AictaatG }i1L3'^'1:l.'.t* C1nderh'?ni.F
:I?u s3Si;1. and emanate ele:"~,`"dd'`d ftoin
20 Li c, I-rw:'ii=lL. in dliscI"cÃ.es but wJi'1ablt 4itE:<Sõ The inconsistent
t11lat:'timj nk. fth~-- small arterial
pe~foraÃon, within free tissue Ãmui;~fers that are vital tc? Ã~io =vivai of
the key cuÃrlneous
p:?n~f? s of .I:.: fl~a_F~ has boen a 3mt#oÃ- roadblock in reconstructive
uirgi,Ãy.
I'he :-noth}adS described lieresn. provide nztt otils to: the
idell=Ã.ticatiSFn and l3c alnow:
of perlbx~t'01s., but also flw~Ã" relative size and dtbecÃtf?n of
,?:z'botif>a~on,,
25 substantial deNigi iia>p?ovÃ:menÃs wb~=I3 they arc: needed, prior to making
an =Ã:-wisiz?ra, Af,Q:-.
Fli1.p C'.lt"`=vabiin, f-?i .mE'.=thod:s cw1.1i tFr.= usei.k to confirm tha,
the key peri.3r"x..tf .i's V~rG're not
izj;..:reZi. dun$if?. the dis:iLctÃ;ta illld that itt`~.' ~ ~scuzrÃ~ supply
to the flap is adoa.,.1~ite; in
addition, L~ie. n'!effiods provide an E5e7jijcL1ve mLki7?s of determining
i:lrw, or fIa; .. dt.4t1I.3oi.`~i
?b} ;?Gi:3:osYs, so 1:h.~~~~ flap :iTtodõfi::at#on or ill#;vF'Ãkatlve
corrective i3?~,:a.sir es cs,3t1.d be taken
<sf? ~:ilttic3~?~?Ã:="~`=u.~~i.''Liy, FiFSa$v, the link3giI1g in4"Ãt7s:3dS
described he:re1;.'# ari. 1seitA in
pvswoporfi Ã?~O :11oni.oring o~.';hetlap, as flaps must L~e ccx?it:s;?uousiy
nic?nit3. ed a?i the caray
day.`s 1`611i3Z.vin#? fired` ~~aJ;l"cf.'.Ãt:iinsfer for evidence of artt;.lal
of vi:ll=>j`;3> th1"o:3h:'Sl-"~' sanC.',,`
only S' *` t ,p t C, f i 4 , T ~`~ . iT
~`~ ~.=~tv is is.Z1i~Y ~.~i~!'~: ~~.n.G~~`a2.~:f5u~.~'.~~'E4 ':~~~f. x~+.F~
~~it.'"~3 ~'~~`=~~' reoperation :t ~~3, rFap,: ~ ~ ~:~C, 14'~12
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
~ A~ a ~~~ a-iv a ~ be ar,v -~~F ` r o ~g; tY
;~ ca
:>~:~.:~ ~7~~, a ~~ a.aa>~~d;r can als~? b~, used lbr ;~ssv s ~Ã~ , i3:a~ ~ i
abia~.~7 and ~ai,ainZf ~ surif
`s lft'its at flap &Avslge7 .17.cxlidii?~ the use of fibI;Ã?(?lil3c
agi.;it`s, vvheà coiz~phcat:oll. o~.-':.x:=n
rF C r: AFii3i',.`7F':Fri1-zF3i:
D..Lziiig c&rdi;Ãe. ;.Ãlte.ry bypass g:affing t;C ABM and ~~flier cardiac sui
ge.=y it is
: 9:Ã13p<}.11iInt to .3.~.~Ss+.vs myocardial ~.r`e#'fL1s1oll. A N IR
~luCiICsLeÃ1i. >Ã'ict',' tffisi> a ..tum?:3.s ia's m t~i;:
i.'iC.a,-I after intTa-ve`,i{}u'~ injt:c~iony s`"v.~.cli a.~M-1~irhyS`fv`no
Ni~~'.vy wfJ~~ld periyiit :ioni.rvauive
ii ta,g: ig oif c'xirdz~~ perfusion.
x
r=F=~~'3'z3d.;~ i.>ts7nC.'s
%
ril?c &b:ifitv iEi ,5.i3C.c3~o the pw.c:-d1yroid :7laÃ1d'.~~" dLtÃizlgI7L4k-
,5tir`,~..Ã"4' ~:.=etn ..~c.: emci{3:
I deten-z:.nant :.~ success, wh:ffieh the pra'ihyroici g~Eand5 zAre tdi~
ini.endeci surglu<zl ia:;;et oz
:iri; ti? i -void,`i d;ng a proced:li'e. I31 some embodiments, th5.
Ti"i.i:thC}C~s c-s";F2 bc used to
f.:.~etect, pw-ah~Toid adeÃromas See, o,g, Kes.viÃ;~y Ã.Ã <i,., BriÃs <~.
Surn., 1969:
56(8)-595-5911.
and :d{ StrEr'~?":s C4nC~3 1"t wNeas.Intu{i1`7s.+tna
Wths~len:: blt i: has pTc:viousiy been tlenlonsÃ,ated to be uaeiul. as a 3
xstial dye of
mere~~ and imquhnonaaa. See, e.g.> KckÃvenv <:tal.~ B.-it-. T, Surs;, . 9'r 1
"'8(,3).2 333-
<I>e P,
. ) i. a i a ' . . . ~- y~ -a ' _. 7 ' 2 . \^ . : and
c. ~?~~`~~#~~~ c~~: ~~~~'e~~ rÃe.~:ir~ ~:;~ii ~5~. us~::~~ to identify
tl?~.,~~.Ã-~ ~..~.~, .>l 4"~~cls~~~:~r x.}<.
other Ui~..?.3i's :li:c to the teildi:.icy of MB to collect in ilshues;. ffie
M'k-3 can bt;;
,~z~ YI C k \ n i e . . ... ~~; ~ #~{ Z~...,.;
:~iil.sr#;\'v?`~::~a s* ztL.=+a~'~i~:i~.li~: Cyr`~a~"!o:~`.ti.J~ ti~ tl?S~
E7rF4i1T~. C .~.a ia?t~? the 7"t~ S 3~~~.aÃ.~ of ::~L C3ra~.t:
20 Systemic %[t3mtt;1Stratlo3 of Ã:h't'., ilyG`. C:'t3.:? z E:t3 bT:' usi:Si
$Ct#.de11tifZ`
1`ie.a~tat.i. Ã'3:n`io: S ftini tis4l:1es that have hi'~y'll uptake of NIB,
ti.g., t?.IÃI,or: of pz3nc'ea.;tic,
G.Lf , nlswinon3.w, .`.Ã~igl-D.
Any IL 1.,Lza?TZ'.=5c+.`nt i.'C7e33pS5i.ilid that is ilf?tE.'cti3b;.t; in the
djqLlL" to be imi~ged L=.r;:,
-11) i.?"'etZ;,r} thoi'aV.:=Ãc dE.ef a, b`Eii.iy tree, or s='wiclil%It:kro.
Ce:iÃ3 bL L1Si'=iI. In gi;t3t S`::.?#, thotit,.' +'õE3r"n.pÃ>I-..,dti
hm ~~uoi~.W~ o in, t1~e N I~_ range (6ft~-1~34)~~ ~~_~ ?) tire ~~seiua, iÃA
adc~itit?~~, a<wo~~Ãs fibruse in
~\ ~ 1.
~
F~.c z~r~.~~.~,=vrs s~~.~ ~: r i~d ~, i~. t ~,~:. ~`' be sÃK~nii~c~~~a~ f{w a
4z~.4~.,,~~.? F ila~~,.i. ~~a~~ by Ã;~: ~~~ ~. ~;
<" ~~~Y.Y ev :~Y ~ ~,~~~~:~~~ s~.rÃt~,~~3~~?c.~ or ~~
i:
F, it b~e t~..~. liver or k.Ãd~.L'~j'~s, i",i. }~:ui~~bi+l. ~.~A
~.+~?~~poun\~s that wohfvc fow.ial, C..rw s
imciti?i.33#:, :..e ureters e3re known ?-Fl. the art, ?i#c:ud#Ã`ig ~rgc3niS.%
filuC3-rophUrer<, L;le Ã:t::?'i
30 conui.3i?.i c3f wwhich~re poly-me4hines. One: in~~or~ant class of t1le., is
i.~lQ
hu~ta'i~~th.Ãp"o, cyanines, coT:lpÃ'Ã.5ed of bi~'3'I, oYa7ole, betvf?:.'h;.r'#
Yt?k, indolyl, 2 -qi.:ÃnZtfino,
;3
CA 02692923 2010-01-04
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"qui-no8ia~e, snidlclaSTe~,. E'Xemp3arv ckFmiSFAuii.b,:51nE9=wude mv6hy~:enc.
~~lu¾; (Ma-t)j
and 4
indocyanine gri,m (ICG), IR-7 86; and ~:~~~~ SOO-CA,
It: so~ne :mbfJ~~In~enk5, t}:. nea: -i:nfrax~d fluorophore has a structure of
~onmuh.,.
#;<
. l. ~;=.;^"'..,
e.~..
=~
: .P
.\;. 2 i=~i,^=" :~ ~õ,l~ `
F3:)amulli-1 I
wh+;.rÃ.`in,,~s valence and :+tsb.ÃlIty piLÃiI7Ã.t,
X represents QR}'?, S, Si'. 0, or i'VRi;
R Ãcpmients 1'1 or li3'w"4'T alkyl, or ii~~'it i)GC:131"Ã'C'.ilces i):"~` R,
ic.Fk'Oil t<3pÃ:thcf, ?:oI,1i a ring
foyethu WÃldhthe i:aÃ';f9natoa~~~s flucn.#gf3w1dch tl~iev are (:~onnF:ewd?
R# and R ri:pmsvnx, i-nde.~~endenilyn st}bstituÃcd or Luisuhst;tu Eed :ow;.#-
aIky~S lower
~r.~c.,; ~:.~ i.~. t ~ or iÃ:~'i.l.~,~+:~r`a~ ~~~~~.Ã<~~1' ~~~'
wÃ.l`~.'3~..#~.:#~L.'`= tL=,
3 = =t~~`w } i{'}~~":~.~# '~~ `ac3c.e~.~S`i=~.Fk ~#, %Ã r~,'3. t~. d by
~~iÃaa{.~A
iÃF~~.,~Z'~, ., 4
phosphate, svdphF..Irx`.":,, phosphonate, a'{:Ã:lt3gt'.t,, ~.^ayd,Ã'i)x::yl,
al`r,:~7.t#.cs cyesno, Ã'::}~ro, c:s,:bt3xv:~>i:~
-acid, oi am1:i.'. or a ph<1,i'1Fu".`Ã.i,eL#~ica:Ã4ly acceptable salt
::hoieci3'}r
R-y rqpt S ~c i iS, 1#Ã~~~A3~'ÃiC~~il~~~' i?i` ( ~%~; ~3~ t:tt~ 3 i'Ã:wt <~11L
t3; ?~ ? ~ Lt]:~.. I~f:i:=Ãtx~ Ãf~ ~~i' '
-r#:ng t:,>'%,h{ch x: .s ;~ Ãa,.hed. sFÃC~h as a fused 1"#R3g, stlÃfa?cK
ph:?:~phate., suJphÃ3t3ate~
pfilo:,p>`sonate,h.1:ogo.i3Y lower ailqlz hyd1:(:3xya, e1II31no, 4=vs-tÃ'1C,.,
Ti,Esl"tiz, ctÃ.I':bf3x4lic acid, or
wnidt<; or a phan-na4.uu .icayly accepEab'e saft tiiGÃ'eoi;
R re-pra,seÃ?ts 111x hnlog31, ciS' a substituted (}'r u1rs4bst.1tET:e(z ether
or tM(}t =thE''.~I' of
ps3ei1ol or thiomhenff 1;
R, A4prosenis, independently for eat.l^ c}c:uuiA:n:Ce, s'ubstÃ,il-ted or
umsubs;:~~llW'
lower iizkya, :;:~~~,'C)i3iilk.L'l; t;yC;.aÃ1:1lkf lalkyl.n c3r34s t:r
ifi~.:~~kyl9 k2ptiorfi1?lL,: sE.1.?:hiltuÃei3 by si`allaw.
M.
p}lospba:i:~? `hoÃac3`.e, phosplIonato, hyÃl~~ge:-. 1.-lydI'C?xL'~lw
aÃe3..Ã.a(?. eyt3i3~?, i~AÃiT'[), ck;ra3i`xyC
-zd, &niide; or a phaÃnna.ceiat:call v acceptable salt the>eiof
xÃc
a:: s~~~~~~. ~=~~`~~~~.~Ã.~.oa~is, i~:x, two t~c;;~:Ã~vcP~.~..;. ~a~'R taken
tt?~.~..~:~` ~f~~#~A a i"f7t.TiabC:.Ã`t:ts ning. ha so'Ãi-ifi`.
C'=Ã37i7i>i"ÃÃ7?o,1ts. RI, R2, and one or both R3 ÃncliItlt~ Sulp~.lonilte.
Ã4
CA 02692923 2010-01-04
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r r~ ~,g~~.{~ 7
7tdfY it~~'~iS +3~.t~1f{.` (:Yi~.i=s) Aftlii.`r~I:ÃgÃi it was pÃ"evÃL?.1sN
kÃ1own to use Ni,B as a visual dve, .he uV~e. of N1113 ::`:
fluoÃSõ'.sS;.L`Tit.fi: nnagÃnb has not bS::ti:.?.s C.g71Ifii=%$i'lfly ilppm
=xtlt('d. As described }3ii`Ã:rl
b.i%Ãi. s3as pF.opertt1':s, Tl?o LmÃas.i<3't] ,'*:sa+ <:l;nr;tb 16770 mm to
{ ...r`'.~~ "lin 4't 3.i:i a i3eak that s}:1Ã:ffs :3..5 a f:I31cti i71-i of
dye C'ons:.'Ã`>I3tÃ:ÃtaC n.) is within tt:Ss N R i't`,Ãn<.õ-i'
'c3: physÃ.t>io:sit:e~~ly .s`s:~~e co;i1coa3trcitl.ons and t~orÃ.;fo3:e
p4Ã.t>`fits hiLi
,h and high
23:Aal to baCkgron:nd citiw t`, low auiofluor~.s;:enÃ;e in huni<mS and
aninials. . his
:~'.:1a1't:cti;I'lst$ti= d1;ows MB tfr` be used as a Y'asL.4.lls:r 4Clit=iilst
agC'lZi., using fl:ist?....si:t>1`:~;w
Ã.:;i3tÃk;I Ãlg 1eci1alÃ's3og JS:lril'is~3~tf y, ~~~? is aeir~'tGC~. or
A~~Ãt;<,Ils s~t~..~I~;Ã=~%lln' into certain
ir. ~~ 1'#~~~ ~k`~~ i3~4?<1Ã~ 5 FÃ"1Ci.3z~iÃ'nw'r~lw ~t~7.t>li~LÃ:c duct,
~?llE' f~i~.~t~~~T1 g 4=j Ã:?3T ~..;t3C:t7 ~?f;3Ãe:ii :y` :rtif`i:
1Ã:6ne ('s;.llf?1. 3;:1.g. VÃS`aalÃ;e`at:Ãt?n of flic. u.Ã'i:.te1'S), liLaÃ`t.
JnyÃ7+. a:'d.ÃiaÃras vÃ?>GulatGÃc :al3.owing
iÃ:"i.Ã.gi3g va, alia, the inZ%Ã}Ãf:ir4:Ã`i1in, comonary artery, etc.), and
~.3,,Ã13c;"ec~~ (e.g., into
beta :.G;i;s; alli>t-Ong L'l-sila.:Ã.:%atÃÃ;n of tt1.at organ <Ã#d
1'`iIÃ'ilÃ}:':i nid Ã.aeÃ'.a5t,si'5 si !tf': a
i1amà $<asa:?c CMg:iil. à ,g., ln6ulÃ:sE+Ã~zis).
N-iB h-as %h~.` adva1:3.l%I~?i.: of 7euread'Lr being approved by the U.S. Food
& t. F~Ã"Ã..1;`~
,. .~..,
At1in'#.a'7sttc1iiÃ3n :3-s a blue dvL= to asSwss gaStrf?.Ã71ti..stÃna~.z tube
pl>Ãuc.n.i~::"i'Ã and a.", a tfeat'iÃ;i'wA
for z>:s.:ahmmoglc~bineÃnÃv: Howcvcr, thoracic dtieÃs can be visualized
clearly offly aftox
4~2 ÃI1~~ll~'~~2~~~:i SÃ~31t?l~' I~k.asi' I z .;
color delea .iÃ;Ã;. But ag +,~tmlonstf ali.d fieÃ"eÃti, the thC?~acIL= ducts
Ca.Ã1 bL dc:eczcs,i ~as>Ãli~ i~~.A3
20 fluorC::sC:ouZc at `mli;;h J4Aw4Ã" L:<)Il"Z'Fltl"at'ioTlsg '~wen
Ã3.sa.ng$h+..~ sr1liiC ÃT3'=fi;:1n tc cla:~iqi Ã;., FtZ =
H aliqale. WIaQTI used as a "Au.,_ dve tonaq? tho.t-~~~ic duct. MB is injic
.,~tefl dlr;.c.Ãav....
stitaà il~ i à t~tÃ:> ;> a I `;"'i, (3 1 :Tl~'~.~ ~Ã3li;tJo$1 in sterile p 'f
iCsjt si=d wat> f: Who>1 tlso-i a.s a
LIti=~roph3Jsi>., and lÃ3ieCted directly 1n:i) the Ã33Ã3T'a<'.3.C dtict. one
:1e:d--~ .a? Ãit:h:+,.v~d- a
L'=;3a?GentfatEi`n a.".Ã the :-#.sstIc of o.Ã1.~v about 3 0-40 ~M,, e;g
it.botit ?0- 30 pM (wh>ch is abo,,i`
25 a ,01,00 1b36.Ã ow(';:=r dos:'v)45f #qjL`.'~ctÃn~..~.~ a distal iiode that
would drain into ~+c 11)j a
concentration bc:wG?ei'i 3 S~ 1.~ M and 33 1 mM can be 1:.;sed. This
>F,a;;ge5t:-, that flÃ.aoreiCe.>ci.~
;:mc:~,?li'~g is n1Gm Si32sifive ffi~,.1 viSldraj inspection with h;.imaI2
c'y'cs or Qoii3r e:i:moÃ'as,.
Doses ci>` a;O-10 mg ~~r. of mefbyIz.ne ~~~~iQ tirf? widely used ,alinioa,ly
'.~~~~ the
L:~'~`~'.ktn~S`.~.:~t ly3Yxnuffi`L:niS.'~rg-lG.3binaemi"(.{, and.. Ã.~iuf.'.in
larger di3sL=s (on i..SiE~~:~o:Ã.t`ti>r of 4.0-7.5
~~~;.:='{ ::M=.
,. s >; for ~ ~g .. ,,7; g> y~ 7 t ~t>t. ~
~> <:i~> + S3~.~;~~.a.#.~~~i`.'f"~,.~:
}~iÃ~i.~.43~Z~iÃ..~'it1'S~'i~''~9t.,,~if~~..it'~, ~:;C:Z:'~3t~.~'Ã. .~.~.
~~a~: ~i~fr.kc:Ã end, z.-.g_
MB :ÃZ:i33Ãt3l:e~i<`ai1.Ã3~i xiÃ3iF1C 7.a:tl.tk causes ~i~~1~IB.'. <~,F~.~'t
i9t ":'~2.f.1 .ÃÃ)e-IS, L;~
m~~theÃl>ogli)b:'me131:a or anaphylaxis. IÃ1 addition, :hGTear% some
1:'epÃ1#tS i.ilddci:ttitfg th:a>
inilc3dt:,maE 1_i1j4 .ctiw:F's~ of MB :"..i-r1 cause :ikitl d:~11atie. For
'xu.Ilapi`r,'~,Qi.ht: hÃgr. Z3<::=...s used for
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
r.od~.1 t~~lection, ::.g;, around 4 tril of :.~~.3 mM MB, a:-cuss,.~i:i~~ted
witli reports ,y,f
iiijf.cLaon :~~temaC."4io.~.~`is. At these b.Ãgh concenaratÃon.s.4, no
;`~liof~.'sGw'3'?cL;' wo':-Ili' '.3i: visible
dE.e.o 4h~ i irÃ~:aE Ã~~Ã`E1~:,t3~ ~3`~'=~3c Ã"~# ~~~~.~ qaÃ4.1'3.cr1ÃI` g of
MB s=tlltssÃ7Ã1s, Tbnis, in K; ÃleÃ'i I the
do`'si:s::iset3 iri ':hv methods described berc:iÃ1 are abi?ut 10 F:ÃÃ1":ee4
lower, 'wf3d in Sonni::
Li'31boa:`~:12v.~+~3s 100 times lower thc"'ln tl:t3se previously used,
a3'.1+.l wrt: expected no``=.to ca::is e
Ã':lliof ti'x_ÃYi faaÃ>:a c:?r adverse .~'i:~c'~"~;:.ii3Ã~.Ã>. For example,
in s~:~Ãz'~~~ ~.F~~?S.'~{~iil'c4Ã'Et^~,. t~:+,. mcth.>:.;:
;nea3..dV:hi. %Ãdn-mlÃstft$tÃt>:Ã1 ofa. solution ;23i:"l`aidll]gat Ii'i]s`:'
U1 % M13. 3w:& about 0.03 to
ME'~' < g OAOS % to 10%, e.g: I "Xs to 3..5%. These pe;'ceTÃtages, are
weÃ:;h::`LvL~ghft,
i,e., a 10% i^s?tt~~.,lt,Z?ai is 100 mg/m.L IE"l gC.FieI"'c :a the total
do>'sc iha. `;`l~l be t3.sed ft1.?.' Yi`Ãi.5st
i~~~?l i ~3tÃt>I3 s is ~:~~i's:$t a 4 ~~~~ of body weight 8t'~34'2i
i~c~113I~: . , l5~'3iÃ:,Il^c;w1;.;t ::<.;= fox
a 70 kg l?ummi, :~.z`d a desired systeinÃi: dose of a(?ot12 I m:Ak:g, i.?IIc
woltlt~3 need 70 a:lg;
~SJ".ich is equal it? "17 3rfl -csf a, 10% ==- 100 Ã?'!g Ãil1 ;iolÃ.3tÃC3;t]:
or 70 1TtA of a I % ::>: 10 t?-1&~An I
:~i~~.'~Iti<`.i#.. I~. is desirable to achieve < concentration "ax"#.'~i.~.~..
t5s:i:a.i. tC? ~~..;~naÃ.?Ã.C< o:`el')L sLit i 0..
40 W, e.g., about '20-30 Ã.cA The coucs:at:;3Ãion e"aÃ.: wary ~ependin{} o.-ki
it:c local
envÃrcruiYcnt of the strucÃur., ca be: imagod c..g.~ the pH of tl-is or the
cÃ?Ã,,ce$~trwÃt:i.`>ii o. paS.3ic]Ã1S: IL`a some i:$Ã1b1?d1Ã2;ei1ts. <:`.
opEÃ1'f7i.3l c+.?ni;+:Ã";.F.3atiÃ.`3n can iie
s~~eTitified, based, e:g., on t1ic graphs iri [~igÃares `1A-D,
~a .<;. rZ herein
It. 3~3~:Ii2'i~2~'~~'; the ,~~:3 ~~Lt~.;Ã`es~.ai:e flfl1~Ig31I~.~
~3t+,~i~'#i~:i~:~ 1,~.:~+~t: ~e.s Ã;~ Ã'C:-alti e
~z# ~.Ã~~ ~Ã.~t:~-, ii-
s seÃ?SÃÃÃ\%1' i 'v~fltr~ lower LÃrsvs of ~'I~. Nle~~?~`lcA~e ~~z~lti= cÃI~
.hi~Ehs. as a l~`I.alp
:?i? bile duct ~3I;i1 ureter 3Ã::d.,;CaiZ?i, tit?d a vascular contrast
a;ons.; Ibosc broad 3nckwtxo3'ts
u7.7'CidE'sL:=C: I?"1e2'4 :?pv.i.;.3x3s for t,:itr:3t?~erata.lv4' ÃP7la~3.Ã1g.
In uddftÃi?n, mCrtily3i?.ne bllle can bC..
used :n cian3.bin>:Eloil with i5flle:i' "ilt.icÃ"e5eci~t ag~:`Ills, s~#:ici .l
as ICG, to pit3v.Ã& multi
),5` ,'4,cloa~g,.th., 3z">`a, AtÃ"-colo fluorescence 7.ÃIii3.g1Ã'ag.
25 N>R T'huc.Frqpf?ores: I CG; IR-r"86, and C PV80 0-C:#4
3('L_s is a d3-:suip}le)i:Ã.3lod hepfiazl2u[h1nc it]dCSi;>.%:aF3Ãf::i:= that
:s F;`,.3Amt3ps Ã'owed for
1`aÃ.d.,;ac and hepi aÃc. f3IÃ1c4:oT3. s:'udi~s. CW800vCA is a cafboxyl1L;
acid analog of
IRDyc.:hic`4.~L)0CW, es newOT hi',p1,'im1efh1ne iÃtdot :''tdÃ3Ã11c iA Ãth
higher iI_uzllltÃ:Sm. yields and
Mo.ar e:Xtax7"-:isEexa o+;1-~~ffic1crt#-s< fR-786 is a ht=ptaf"21e$hI.ne
?,ndi>r-yan3.nic with E"Ã{? ;;u3pho ibit Csn,
30 ,~~id is att .:m-zo>.`ÃeI s= hydrophobic agent. 011 the other hand, CWSOO-
CA is a tc;sr<A-
:~rt:lx~*IoÃ?atL d 1-k a3tt3ITieYhÃnÃ` indocyanine, vvh1C:h Fni IE a Cs its h
vd~~ T'~> htii+=ity
~
z WSOO-C14 ~" l ~``~' F~ Ia~c:~: '1'l:~c, e~~~;=~-~~.~x~~~ic t~c>id of
I~~.~~':vc ~:~~`
pÃ"Y:paa(:3.A :\^h}daoxy:au\.cft~,,~~"'Aidc Cst>:., Ihy
16
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WO 2008/076467 PCT/US2007/072800
hydl'ct~c~^~<<i of ~.}:e ester 1] water at pH i>.:?. Thif.`a fis xi':.etr.:s-
.s'ux,?hoà atL.=d h.tt?'s3a'TIÃ3i~1t:~0
i.Ã1docpcai.3Fnu with eF?t;.S'sTC7ii ^ 800 ÃII7. After t11t',tvi'ni3LiS
injection it is t'ap1sel`4'
cleared by: 1) the liver at~~i oxcre¾ed. in.Ão bile wnfi ?) tht. I :.~~-kc;~s
and exerctcd
urite: Tatus., this dyc is u.sef~~ for ima.r~.ii;w t2ic biliary free aiid
uretUr..s.
w ~~'~}' ~~ Is t?A~~ 111c. . CoÃ:amcreiz-fflyr available and FDA-Ã.:ppruved
near-
:nf;ra:c'd hcpitanic.:~~ine indocyanine fluorophore that is dÃ-
sÃ.~~.~h~.~~~~~~od, >3tlt::~
3:1I)eZ;t1oJ3~ it is rapidly t lt at`eCs fT i3i1~ thG blood r? S' ~ he iiv` ;
blit is i;taly
hicffiowenÃly transported rnto bile. ICG can be used to xn<~i~e the
s#.raetwres
described herein when adm1l?.,Set'ed by direct a:"?ject"i}t" or
cadin:.tlwdE?Ãl of t`]e
i 0 ~ ~
~it.Ã':~.i:~:i.~2'~i.~..
;R ;86 (Shuna,Aldrich, lpc,): Commercially avai1abIo.non-s uIpho.t:~ed
nea_: iFttai3t'a`,.'.d hepI't3.llie~.~~."$lile ikldoi.:ywi:ine
fll,lot;'.i?pnC7Ã'e. After :I1trad`ve3tous It.~i.C;3::0:1.
".t is r~:pidiy ~xt`icii i.i. into 9n3ny tissÃ.iGs 3ii. tirc ho[ty7
especially the I..VOF', `r3.Ãat: I4
inL.f-ficik afly 2xc~n""Ported into bia'~~.'. U~'"786 cait be ~i+9ed. 4o. im:-
s. wL~~i~
ti~scrib~.~.: he'~in when admÃiiiStcre~ by direcà :~~ect..ion m: E_
znrtul.aiAon of th::.
~'Ã. ~." .onirnerciall4f available tetra-sulfonated heputÃ~ieÃh>nk:
IRD
indocyanine-type NIR. fluorophorc witb. pe,& abs~~~itic3ti at 772 tnn. and
~~oa1:
~l 790 Tim. HIDyO'S Ã~:~ar, be oso-d t~~ J."Mage ~~~e. si~..~ucturcs do-
sazzbtid heroir;
20 whe;,i administered by direct t7~ectAC~ii or cannulation of
t'hestruc.EÃ.o. Si,e, er?.,
Zaheer et wL, Mol, 1na~;iÃig, 2002; 1(4):754-~.~s4.
IC'G wid IR-786 are taken up almost exclusively by ttY.o liver and
Srwz_,et:~.~d only
~iieffic:i::naly into tli:: biic, bi:Ã CW500-CA is readilsr Ã.ak..a-i :~~i by
bot~i kidn::v ~:iid livcr, a;?<l
%2SA3t3p'ie(a >,iit:~ bhl~.`~ i1.id :ÃI:i$ie, S1::1ce flit l"a3a~Et3-r
c}.]e"micil. d.i~.~o"el'la.?Ã` aE`non~.M~'. Ãht'sf' ntoiecWoJ
s i .~~ =.:~ i~~:~ ~ i~~~~ of s:.Tl"~r~:L7t~ ~..a.~i~tn? and ~ *:IL ^. ~ ~ ~
Ã.=sG'I'~31.'iI ~ herein, it ~? t C v"k .1~-:.~~
~~r`..: t~~C, ~;i34~i:;i~ ~,`?Tl ~~3L.= I~:%,s 4 #.'~^, t~.:~,SvI`afa:
vxpC.+-'I. Un<3w, t3:i3y t't,'c3F,'=<IIl{,.:3'ed fl7.i.C?d'op.`fore will work
as b'J't,?ll or lbi3te`~.'t than CW800-("A.
r t'. . .~~~..
proti ti4 d h'2, t it =s fi?~.ior~,d b~~ ~ the kidrY<= b1lt~~'r i3 For " r
lve:E. ~t:~x`3. ],~i1"i3l~~ :, 4L3L'~< ~ :~~: :3~'t3~,~~~,, f ~ \ ~ =~.s
, , .
i:~c3~1 .~a3 Sta:iphonats:d.
i_i~,.'~;3:~.~~.4 f:='sxFl ~i:: 1.E~4~, a ~i1Y~~"'~:I:z13.`.~;`~::; ~'s3~~<'
;~ {F: tt~3e 1`C;rot.1~~4 (145~~d~1~.~; ;~~ .~~IÃ;: >~ ~5
I'oilsC3s3.''stble tct t..'2pC'.ctthclt peklt'r.t^^ :md
hi.pt~.mS3.Tlph(`slla::ed dC:T'tvstt"z'v'oti shoLildithiC~ kvC3.Ik, i?ald.
n~~~~3~ L~ z7.~'s'~ :? more ~ ~t~?#t~ t32]~L'.1~ SliÃa~e t~3t'lt' excretion
ji3~'o bile E~:tid urine iS Q ~,p.~.z'~~ to 2-3~:.
;r:tL?~>rv xt:~~ =
mo.e rapid than i1ic t,,Ãra-su5phonf~te:=, The tr.Ã-s~.~lphonat.es will
prt:r....mn :ii an
i.i:i~r?.Z.ion !Yb'iChzc~pect to di"aid tetra-su~phonates:
17
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
C3 AdA#2f 23;i1riitFi3#1
in Ã-b:; ~:~.edi-nicziI st.udics described horein, NIB h:~~ bcori fÃiL:i-izL
to be ,~s'-efu; i-v,
~ im.ij;ig :he bile-ductg uretcrs and paraffiyrord giai-W. In som.. ct~sc.s,
los:, than opkima~:
Pe? .oi-t~?ance was f?:>se.ved in the ureter. This is becauqs;, laku marrs_
other dycs, xÃ;.e
1'~~~~~re~~en::e ~.~i-operties of meffi3~Iene blite a3-e sensitive tc. the
~.>E i: of its x;?
iic..d,
'1a ccmkditzom'~' ffic abSt~rptii3n and. quantum 1 effiiit'=l~cy of tl14
fli1:i5ri!peliiie dt:c}eei~c;i
40b'Am3t'.allb''. r.,hui, i.l `f1i.33 ..a-sG4 s3.Gt?ve b1.?~fe337g \'.+z
Ãlli: ?il Si:i.~ y. p..3 43.i":h a iin.i,aiy
~ ?~ t> ~~=~~~ t r ~ ` ,~ ic- r ~_
i 3 i:l~.c>zl~c an ci~,.~Gi?t t.~ac:x rc~3SC:'.~ t i'= pil z3f the
i~fl"~E~.~~ ~ i~,.f,., E'>C ~Ft~ .. _i~}3i3.?s31:~.: +:?~
~=+yt ;~~~`5~3~.li~il~e} '~C1An =~i1st~:~'~:=~a ~i '~~,iti-i ~iG~t~.~l~ t
'>~:~C:i S. 1~~. ~ rr=' imaging
ia~:w.
Z~,~.~ is i:::3-;3[aõ~ a h~;i.~Tkz~~ aEa.71~~1~5~.~=1'"~~+=,~ for 11,?,,y
~"1C= `x.re:'ri"=~ (e.,t.x,., b~ L:,f3 I ?C'et.?7.ylZ.'`ile blE3C%).
In St?:tl?C3 iun33od.itYae.?t5, the methods C+`,'.~sc1`tbd:"li het"cif+
:Ili.,i[I+`..fL. ffi'=
~5 of ;x:-~eof mo:i`c ~~ther CoiA?pcy-und5 that ei.t1~an. c the peftumimee tff
tho dyes. :ior Lxan~~~~lo,
:
i~t some c.~~>:~z}dinnunts, a
EXAMPLES
The ].. <'entloE? i4 fUI"theT deacrib:'.=d IL3 tllc. ~~.3flowing Uxi:m(?les,
wli1L ii do nt?t lIFrk?i
the scope of the :h?v`,~`ntimnL3L>t9ci.iV'e'.7. in ~~-le i.='l<~~in':f.
This F-IX.a.,n& dese-tibe:~ experiments that evartoaÃed the ~~~~
~luc3res.,z::'tt: ~roper6t<s
of:~e-vea~~l d5<`cs >m;ud~~~.~gmcthylene blue (NtB), lCG. IR-786, ::ixi:` C
W800 CA. L~ing
t?~3~t~;.~~~1~ z''~~iii ~~~~>t?Ã=~ spect,omei'v m'Id fll1CYiG~cencf: quarmum:
y-wdd
25 `-`L?:,'s3fbaacm:= spectaC xiI2Ut15` wLiS pe:foT aned wa1:h a >-c:?:;
padl iei? k':h t _3ar"lz cw#: i=t>t: in
~ ?
a USB2000 :k~.~~.i`--Ã>pti~. ~,..~~:~:~~ot~~etc~~ #:~~ce;~..ti Optics,
Dunedin. 1 I;: . ~ ~i~c~F~~~.~ll
spd..L;=t`i;F1?~try `=A"'as pf;ra%mmeia Li'i.tli a I -%::nn paldh Icngth,
>?4pf iS.b<C: k.rw'.?l:it > yIi;1tL=
o.wett.s; (Fish:.r Sc~entitrÃ:.2 llmmpion, N1-1) .i~~ a FdR2+:100 fiber~optyc
t~.~oor:-omet<:-,, a-id
xf:mr-Way cuvette holder (Oceiiri Op~ics), iising a 655 nzn laser diz?d.=
:::g~.:_:
37 source (O}'CONA O.E'. fim,. Xiamen, ChiI?i?)tv~tb ?i:a illT7im: output lt
ks tha: mW. Data
ac.{uis.i1>f:Fii was performed with a computer us.1ng zlli 00:-Ba5e321_
sp:~{:tmmcie: t?pi,ro-Ii11:$
sc5Pn4a'e ',,'`~a'i.;~~go (Occan Oj)fics). 'I'}li; spectral riillge of the
UÃ?B2000 FT
;. ri
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
iuas fion) :. 50 to 10100 Ãini with spectral resolution of 7.6 t3m, and that
of the H R-20430
spectrometer was fi:?tn 200 to ; t OÃ) ilm ~i'ftl1 't' speC~tral resolution of
6,~.i ".tm<
FauifF~'..-u3'.oric'C ina`L =i.a`'~~aty wasnnF4'.&suF"LJ'lf.iF,~ ...~Fc.(~
soEvut (F-BS. EiBr.~.~. arai-nddmnC.;1) ai
i,ad;:h ca}F:IcSUS3trat.().?~ o: thf..' dye to dde.Ã'knzI1C= the offei~t of
the moihyl$.t`L bhx cE3ili:.CntraftFO.:1
A sC?i'b's xv on
;m
'- iÃ3ia': ~lEaE?F'es rriGb'T1C=ld:. >~ lu<324 iCi'ncs quantum ~~~ldf of m
PBS, FsS, :i:'.Fd metht1.T]ol were L;ale'LitatC'.d using t?xa.r.iut 725 in
k:"iyl'G.nw- glytvoz (d,;umÃ>t-mm
L'iela? : 19%) as a cs3l?brc?tl.ffin ;':$amFLi>Frd, LFÃ:dC=F the
cE?d3dtt1.oÃ.as of matched absc3FJ:::nC:e f-3~~
In~th~~leme blue xz:Ã Cac~~ ~~~~~~ent.
MB :;F <. ::,~~~~~~~ m.,~~~ecu(e (M.W: 31-0 I7a) caiio.Ã-aic thiazine dye that
>s.FDA aÃ~prowt:(;
d a~l i,~t s~ ~~if7: i~:~~ 1Ã,d #ex~t~?Ã dye Ã~7,~~3~'li.1F~.i In t+13i
Q:'Li1Si: ,~le, ~~~~ ~~t.F.Ãt.iFa.~ef~ h'<?i"#:F:
N,. .~ne PhtiF'ma l_..c. (I'arwnus, NJ) was diluted Ix-~ adequate:= wns~~um
of'pho=Ã?=x
salinw (PBS) :o obta:i:~i 1, 2, 5, 10, 15. 210, 30, 40, ~;uad 50 ~i-M bof<3,ca
use.
The, opt ca] pÃ"qpeÃt:45 of IMB are Sut'~lmt7..~:?.f:d in TtF,Jle `' ffio
a71oa1F L=:tLF:Ãla..li)n
ooefficient, in~ i ~.~S is ? 6.000fi aml fluorescence qua~~l-uni yield ÃÃ:Ã FB
is 151",q. >hese
>otF.iE.S` indicated i,ha`. ~~'Gq flu$;rCsi:eflef::, which has an
L.~L'~t#nc:.IiJa1 ci`. (;ffi+:,I:ri?: F3nd.
flut?xoseence quaF~~turn yield in FBS of 168,000 ~~id 9` N. respectively, is
:abmu~ 8 times 11 iam`?,t.? :~ic' ..,ii; ~~'i#i'!1G i3~L~C`.' ~sIF..F.~si"L'
aa t 33i:u p E i SF:'>
Tab~~ ~ ~ Phy~icW and Opt-ic I'z"Ã~~erti~.As 0~.~ M~thylent Bli-~e -anc~ ICG
------------
....... .:....... _____ _ --- ----- ..... ......._. __ --
Molar ~~?~.
:~~s31~ ~.xcitt~tic~ll ~.ri~~FSSIcsr: `' `}~
Q~A~C~cZ 4im
EYtFkT3C1101A ~'~ avelG1'1ghWags4leIlgtil YÃel.i. Ex, <:=r x ivL~~.~~~;. < <
M.W. ~.~~1~'.?i~ ~, `{3xISZ1u321 tc 12ti7itFli~i 0
r os ~ tt3 tt?
(:~~~) (~~-Fci~~ } ~9:1~~} (r ~~l) P~~S Ã~aAe U.;Ãne
PBS (~ ~~~) PBS (FBS) PBS (:~BS)
[Mtan] ;Mtot:11 rmtolil
~ ~ ~ ~ r ~')
M <~..th1l ~.11e ` 715~~~~ `~6.. (666) 68u~~Ã. ~5..>t ,~ . 5~ < ~e ~, (2. _
~3i::Ã~.~ ~' "w (86500) 6 5
[715Ã30] ~ [681] [{>},5;~
I i 3 .............. _ 1s.7000 779 (8~~~} ~t~~~ (811) 1 '9)
(168,000)
2 u
a.n addition, MB -flufS:t;~cmCo iN wlth'F12 thi, NIR Eim~e~ .?Ã:d has about ~i
hw3C>>w:d
::~~a:~Fa~~:te, c~.affcà e~~:. e in .xrt3v,l~:t~:~t~3 ~~()Irl ICG and 1
~Ie.Fvt:tÃi e . This
diffc-mme should pennit flae 5eparF,Ãzm of these fltÃc:>:O,cÃ;nce with. s;ur.
c;~,Y,>.1
iÃF~ck~~ik:g '~i,sa .
P9
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
C3p;d
~itnhy~ene blue is in c<~tfflit3ritu-ii wat~l Iettci~lre~lyhwn~.' blue,
tlwlatw be-Ãnf t:lewrr.
'k;.Ã it.aa4: s w.`ad `1oRi' flus3aesfvent. 1w11i. C'.S?nvii"siL3n.
Y`=.?etLi'oC:n the two is .i:i.iÃ?x and p='l^
p~..'..i':de33i, wi~i, ::.igh ptil 63 fciva3Ã'ÃÃ3g. the Il19ort~~ce?`kt
Ã?1C=ihy1iTle blue i:Zrm and h-tw:`
pA S i : r'oTillts the Gl.t,`.ur, colorless, and ni,t7.-fl 13.i?Tcs ci fi:
roi`in,
~=~eCa~3d..:~e bbue oxhibit,,~ complex fluoÃ'esce:.f.~~.e output, Wi~~l peak
;f?w;~s~~y fz :f~1 ;:In 1? ~~. ÃIja.v`~31~:t~ii:ifi' ~:C1Ã1 [iltr :tiC)~?s
li~;`~~ t!~~ uM. Ii3tCÃas1ty is Wiso h gb3 v
ik,"',z:nd::=na t?f; C,1Ã;.i71#.e>'s1 ~,`;:'vi3'o#.ii-lf'.:F'at, with
}il.=.ti~-.~.~.a output in i.Ãp1d-ikà tiÃFr'.7_iõ+~ F 3:~=f3,~. Ft3I~
~~lo, i~,Xpkmm~.~ntshown in.Figpire 1 A, p-tir~.~ m~thylenv bIre powder was
distiol;'cf; iÃ-a
ph~~sphate-bti~lered S:.2~~~ic (pi[ 7.4), 100`f~ fetal or <Ãb_.o.ute
inÃ,,,".rA ioI, a:: tli.-I
CtJneer%wa if?:.: shown, EÃni^ flui?r`4,=4cC:nce intensity (aÃ'bÃ.-ff awrf'
vs". al.Iey) aitc-iksLÃr+`"~d.
Metl?vic:Ãie blue exhibits complex optis:.a: behavior. Its wavelength c+f peak
~~tislla~:Ã:Ã:Fi3 C.;?S,:~a~Ãi;~l#:'f`3~'+.. (i.e., i~~?4E:.~Ã'+3e:..~`~,.e,~r
is C~~:"g?+.:~Ãt~~~'=."ifi. `~?Ã: ~:i?lIzt.Ã1t;t1:tiuT; and (.he:I3iZ:afl
01:vxroniaaunT (see Mg. 1 B), a;x is its wavelength of"peak flui>Ã-esc:enc..
etr:i"S:z?n (I .g, i C),
For fl`_:s C,xpei3.a34nt, }.`Ã3,.tÃ's, Metllyli;2'1e blÃic powder was d1~-
Notved i:3 PBS, 17,BSa or
T'nGthÃ.lno3, at t17z'. k:?ni,::ertttat:oT1S shown in ~~lG'4; I A-C, the
spL=CY3:u-n of .lts eabsC?;E'b.1noe
~~c~il~ of ~a~~~ ~~~~~Ãi~i~=:I.
flizorw~eu.cc w :~issic~Ã. obtained, ~~.d the
~a }
~T <-(i~t te.~l fluorescence <a ~ ~ )r. ~ "l.=>ld of ~a~t~t ¾ ~' ~'t ~ylT'~~
;~e ~~a~.~fs. ~` 3 ~ ~i:i . x~~:23ir'3i<f.l~~C.i t ~ i"3t t`A'~ w~ = t
y~~Ã.rc~k C=~t?i
}~`~= r 4~i ~w~`'":t?n ..~~~.>
``, I A) and pc's3k iÃbf3C.ZI'baFAL%t ~ ~ve_EE;Ã:?~th reI7ia]t1s sto+`_~~ (I
B) in zl'I'Ã:hydvta~,", wide ;.?.{?;;e i$f
2t; the dye ct?nccina'r3tif3n5 aD'aad pGak fliÃozeu%onve 1:1aStssIoÃ1
Wil'tielLÃig-thd{:xes IiC;( ..1cÃ::'loaie a=.
bÃ~,~.~,hLÃ' ci>n-c:.::;Ãati;3ns (IC), allowing ~'C.31' aCtjlist:znent of Ã3-w
C`i?4o of MB fo'" Fr`..Yap".i=fg t{3
i3pi'm:i'fc si
',.'ii_:a i9 background ratio.
FÃnafl~,; flic pli dependence of NI[3 wzis bv diss< _ ~~ingM.3 ,:tf. PIRBS Ãp;-
s
Ã>~, ~.5e:=a~r~~ic:;'t.sõid, x~C'~, a.-ii~i a Na~-a~'0:~..~~~~~?~e~'od
;~~3lt;tioas. As vhcz,~~~A }~~ i i~= ID, tl,z _
. . . . .. . . . . . . . .. . L~.
r~
2;; ~ 1r~3;~:s;~s~Ã1~Ã ~ :i~> no ~~ :iaC~es~=..s ~~C fi:= in acidic -a-~ the
~ StAt a~ttic3~~w= and significant ~:~t:i~.~ G st;i`,~: .of. :t. ;~iu=
s . .
:nienla, .absoz,p,i.0-a Often limit the usable dose range of fluorescent
;.oiitTas: aguc.ts: Thõu:,
the conditions aliil tJnCeI1t?'~~tion can bo optr:nÃ:zod:
_r4
:30 IN?R FiufrCi+)?'þJr'..'i: .l.
R 7 86r and 0=VY00=-"
IR^71 86 and sf."+dà [;I:.la3 i.CG (C'x::"d7o5E4% wl:."=rE:= pÃ.1rchascd
ftoi?:. Sigma (S:, Louis,
MO). HUM vv.~:.~ diluted to 100 gNM ia3 ~~hospha;e-bufferec3 salinw t13BS;i
,,tÃ,splen.ented
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
with c{ )l.1{> 2 !~#C/ '.. a~ ~ '4 A ~~:).4'rn~ZF~ ~~ 1~af:4=a~ 7 .~L
(1.s5~t.{2 ~,y j =~~Iy)5 Sraq{S >0iL5'~W ~(r~((tt.~2 }~`!t.L+~~+~~ .~A fi\ i e
L=i:~t~r5"~tF'( \ 4a \p^ }t tt-~cYiv~~ ~rr~ fi } !\q 3'.
A~t~ L~~~\x.t 1 ~ v F~:~~i ~i ;~ ~
<
e.Ii~:~i?~~Ll':-c
CW800 CA is the carboxylic acid foant ot"IRdyJ '~ 800CW NIR dye, wh:4Ii was
obtained as an NHS-ester from LI-COR (..sir;::ola_ NE) <~.~~d hydrolyzed ini
Law,i-teabL3sc d
:_... , :
:~~Ã~C'=I'~5x`,i 8.~? ~"i~T' ~>I~:i ~~}'~Er at rf3L~i~~, iC~I~1
~341'u1`;:l~ri; ~7~'.iE3:t`4.` ~#Ã~'~7z.'.a$.?ir`.is by ~IP LC i:+n a C 1.
c~f.
5;.:?l;aI711:Ã.
`Ã. ~.~~~~~ ,~dss:~: ~f?l~Ãis-~Ãa~ ~~~I~~~~ ~~~ ; ~ ~~~~1 ~:`~~FS~i# ~',~
~~r~r~= sÃ~~Ã~:~~ ~ ~;3~~~'~t.= ~;~
,<~,C in the d
:ar,- ICCI and CW800-~~':-A wer~.Y diiuted in PBS boabr::~ ~se,
I'A3L?= 55pt.ici.Ãl P"3pw..'tÃes of these contrast agents are sÃi.
`1r>.il:Ã'Ãzi,k in 'riibl~.' -2 A:;
3 0 three iflu3.3Tophvros have similar peak absorb%3ncc and C #elÃ'.Z`SIf?I`.
;3 :Ãb`+,.lÃ.i: :h4. a.:#t> flae
Ãiu`t`sÃiaml 4a~,'=sd of CWSOs~-CA w;3:s t[lf;r l3,ig}1e;it o. the t5'irce.
I'able 2 --- .~hy:~ical aaid Optic Properties of NIR Flu0rophoreS
~.._ _ < < ------- .. .......
~ ~~f 13cak Peak ~`.`,~,.Ã: n um, ` xer. R.. :.
Ieve:
NaÃ~ie M.W: Sulphcz- Absorbance Ftuore .erzce Ylskld k3 =
6 Ã.3PBS inP~:-~s Bi~~~:;;.
rr3~f.~ZZ.~~. , in 3.~ #g~ .~3S (z~'.nm)
r.~,
~'W800CA :14: 4 775 .f~`~t7 9
;a`a~.` +?8t`4 584 0 {~~~ 803 3,3
ICG7r'6..F. 779 806 :E
eifusa7~~r~ L'p 1;
s
.2:
IT,F~. An~~~~!&gjZtji'j.: LÃ.i M-voz~..._... ....... ......-,_ardif?3 P
ggu~'YDa~v1Ã:}r, 300 g zats obtained from Cb.;rles River ~ a-bo:at~.~>'ies
Ma1~ Spsa
iW:h-n:ing:c~~~, MA) w4r~. ara~~thetb,.ed with 65 zn`kg H' pci:tobafbita:. 30
k,- adi.FFt .:enltdle
1 .. & MA) = ~b~.'Ã~. > iÃ~~~~F~~:~. v-. C with : <`~ .t.~ rx.,~., };:
"? 3r~~.~it~,-~i~~ : ,~ _.~~=~. ~~~;~~~~:s s'~ Sons, ~-~:~t~~es~, ~~~~r~.,x ~
:k,s
lEs:~~F:~~iSL=~zii~~ r>#.Te'sl.4 ~ (Fon Dodge Labs, l'oÃi. D\?dt.rC'=,. IA).
Pigs wt'r4 ia3ft:bated.::tz3.d
.t:i#ÃÃai4`rÃ.:#*c-d with 1. 5 'y 't3 154,1fiuraÃlt`.,
Lq a1?L=s.h..`.Ãi.eÃ, rats, 0.1, 0,5 and l n3gskg 1013 we-re, iTljecÃ.eil
mtraveaf+ua>y .nd
~ to ~. ,=3 Ã.lltk,
+=,, ~=\
thL 'p images takei-#.d:ah'ing 15 r`a.~#.Ãl at-'ter injection. Signal tt.
~~~ik~'' ~ ratio (SBR)
(the S:liat tÃl of 1 ftaf3E:~~iii 1` ~~at S~=~^eiÃ1~ bÃw3.ch of coronary
artery (LAD)'CC:lafiv,' ~ _= :3?::i of
chest we''.:ll. and the sÃ( g~,~,al of myoca'L3Ãlm relative to that of
cl3(l'~t wcà Mw<3s q.Ã:$a:.]fii?d
20 :zslim;; t[le stiÃlle region of interest {ROD through etitire
c?bscrvtÃ.fioÃt In ar?es.h:.l::r..ed
f~Q i \ ~2' ``l : .f
: ~3: ? ;.i,J__ik,? of i;<<.L il?C.,tt1~F'3~;#(. ~.?i::E~= (I .{~?
,~1.121E7l/~i:g' was ~lY :a?~.'wa;e~. 71't%ai34'i3'tCr~z' ~slZ"iÃi.E3 l3f:t
::fl.:,
s wer~:' obsc> veduszng the imaging syst::m descr~bed hc.=r-.An, 1,~:r ~~lul.~-
Wa:Ivea .l:gffi
;iean
21
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
>.f GlorescE???ce I.Ã',.3agÃngy 0.25 ml of 100 6tM IC':C.r (75 11t.:-101
Tl,~~,I) MI` 1:e jC itOd
rats ai.ici>).0Zlntg,`kg KtG ({ ~~~no1/.'E~.g) in pigs.
neu-in-tra.red tniing ;;Y~~emus~.~l in this and txe following e-,"a.>tple5
IlaS
:;S.t;Ã'i dE:st,iii?i:d pTov3oÃF~ly in Ms3i.l (JcC;, e.g.. Do GraI1.d and
FT'at:gZ.oi Zõ .~~C;i:hÃ2ol. Cancer
Rks. T~ew., 2#0.553--62 (2003); U.S. Pat.App. :f'ub, No. 2006'0:05509to
F.anL,;i43~~i ca al.;
[J.& ~.~at App. Pub. No. 200:~,=02850_>8 if3 Frangicrn: U.S. PaL App. Pti:b.
No.
~~',005, 00,....0~Zi 4~r~ ~,., t , .. and ~,~.S U.S. Pat. :~~ ~. 7 T. PZ. 200
` r;a~'S a~ `2 E to
~.= ~~ a G'~¾~Ã~~~~i~-~~ r et a.~.i~. ~c. . ~, t
FÃ a.igit'sni) . T, i3i: hyste11: WtZS, 43i.t_j Z.tStÃ:ta tO ',Wi:i)l$.@A for
~`illc Cipticiil P1"f3_3t:F~'a.toi C3fn.wth'?riE.`:Ãi4=
b1u
i~ I_~Ticfliv t~~~ ~~f
_J-a soiirce of the orÃgÃnaI system is composed of a,~vlhaÃe laglit s:? wrc.
io a.~t ^z~z~l 7 ~. .:~"f-ha eti"rarF;.Za ~hy t ;lt source. 2. il~"'.
su1`SS1+: ali i~~ Ei~Z[i I:TF11L::;i.~ii i.~IS:;L~: S4Et~1 b`Ji ?~
: w+ Ã
:'X..N a~.Z.''.3 itaea:cQ i"citSY Ã.s 5 -niW/:.'iI~?. White light and
:xi.y:i,E 3IZfiai`E:d images were able t.? })e
t:avt..#I'ed svA02Ã.i':3tG=]Si [I.sIf3,, a dÃ.C%h1-i)ai: filter cube 4i.:::
and these F;.Zimt?F.,s cÃ3il`si be I'nfi,i-.Lii..
;,maH;es '.1Q1c captured mZd rt;~fresh~.''.=d 15 times a second, 1 3iÃS
cup:$t'.1l1:y, w1t'll 1oCtn. .I#:; c.:.:3Ã3
a\}t1_f ro`.ktJ 1.LS131.¾io1, and a cardiac gating function OYvSEt~h
prCfV=2dV4 a clear 2ma~e, of Ow
ht'aft ~,n:}Z f;vv I7Z=L?:'1i5FÃ a4'ti ftif is by 4yi':t;:blaZt]:Izit3g the
ffiZZ3kZ;Y of 3iZli:Ãge capture wiflh c.A.ti{:=Ãat%
aTid :~~~~~ias.s tFzrc.e hi)aMes at tl-Ã~.4 same tirnV), allows .4=za;-timc,
precise
Ãf.fiontific+.atior, }~'3 .`h'~v anatomical structure.
.
T}ti. S4`sI~,'Ã43 was #lx{3f~ii14'd. in 3 Ft'i'~'aaE3r i3ki."~~~']~.t1 MB. :
Ã1'.~t ¾white 'i~f~t,c x~:~.~.;.
. ,. . . . . . . . the . ,. ~_ a. 4 's r:w.. ~,
: Toclino1c.~g~A, Brattleb(.)z(s, VT). Se.ond, 4pa:irs oWf)
:-PA, 660 .ri.m, LED,, wc;rc added and fiatcA'r.~d w~~i. HQ650/45X ~e1x c~
t~~~,~ insread' of dr:te
i`lcai" il.:f e'r<ti`+:3d light source. EKi.ltii:taCi.il fluence rate w1ffi
this ~.`ow 3.1glZ.:. source is I i2:w13;,>. "..
Th3:'d.> the diihÃ'uic filter and emission {iftC'.Ã' were changed tt's
680deAi' (i,'hÃ"o:,>a.),
HQ (.)(Y':~~ ~chronia.)f respectively. T1::i5 al.czw", eil~cttve sc;paration
of white light and
fluorc~scez.t light eoaningftom methylena~ ~it.:, tistd pr;~-vidles color atid
'r'_: .` separately, and si6lows display of the iivo ;i21agCs wItl1 a
me.t'ged. image `~~`1nwh:~a.1eo..Ãslb' `,Ã;:
:'eG:]-fi3:Z.t:.
Ax3gg?:g{
~
;'~~.~tc?i~uur~.~scence oiit of rAoTrnally peÃ:Fu,~~ed rat hearts wasminim<tk.
<
.~'~ ~z"~' ~b'ca~:i~ used to Z'14i~ic~ 1t'.e coronary arteries, bi71
~3l`.'7'l"~7~Zl:T"`~.3 _,Ã"a#~~~::li.w and cE33'ÃFrli F'y
310 vÃ'#11 'svvxe d"io` dtUte-E:=t:~bi:: $i4f.iZgNIB in 1'i3t:s. I#.iti
awzc.~4a: i,~laT (aÃ'4eÃ``s;Z-l) sIaF.I"Ãi:I:k wcF'e-ni>t
dÃ:teett:d witb ltZwÃ.:i: d``~esof F21ethyleileb31dC: (OA Ir1z?+`?ig); with
lsiT'~.?C" a~l?~~:3.1i.Zis (0.5 and LO,
,n~,~=~u;¾ coronary arSer.4s c<o~~~~ be detected. Met1i~~le-tie blue is
rapidly ,<,i&ri~.~Lftcd lro~m
y~.
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
the b1oo~~~~earn, to myt3cardial t, :: iat ::;-lud;n 9 the nwoc.ardiw:Ã.,
which likely expl.~~n,",
w.~.~y #~~ic uoi~~~iarv verious return cannot be ~~~slial~zed.
A F;~igpxai f;F`ovn '~he he&rt wall was detectable in all loading doses- of
~~~~, Whii:.h iuggosÃed that MB e:~#~ be used as aii optical andicato> <}f
rxys.>cmdiai porA'V,SI:?D.
'llii; #iat::nsFly wa3 dep'4aides`~i.l on thi; loading dose of Ãi,tethyle:5e
bl1:1 .lowev+,'.z', the deer .."i:so
of the fluf?it=L.3:ce in the cardiac wall was I:Ã#1ea.Ã' and the Swt pe was
ci,F'.z.Ai#.r#f and
independent of loading i::osÃ:i
is?.1 a i1t;;tn.:iliy PC, ai.FS~':~d pIg'ti hLil:i"t, aCitol3LFi3Ã`eseF:nf<o
was :nf#ifiT3al. Fo``siow'#.ng
:~~JL~t t1i,ll of 7~~e'i}l~'l~.3~G blue via ~~[ & xai:~T?~ilj 11
"ai~~F~.F~~' b'3i~ ~l:w;~~ ~ ~;3.~~ :~?
%i? bavkgri?:i'-.i"id (S;,.skO a12aa:;''~`.s of zi3',te1Ãtil blood tl`+w kIl
the beafi.IIg , 54-aà e. 1~"cf'E'. 6b~tt3aE
t`yp r~.~,; )r .3nd. sub.w~~~~i-iily, thomo~.:~~.:~ious fluorescence O>it of
the Whc:Elehea:rt Wa1l w<ti.,
i3 _gt; SBR Za'%ibl3lit 1.5 (Figure 2, Irt'cldie). WheF7 aiv.."wta~~
cFTte.F"i;:Fz3 o4.O1;#s#.{.SF:: ~'L%a;i
FA.itF='oducvd by ligation of i.ip'l'vripS1Le2a1 diagd.1na i t>r'tFa
~Gfa't..Z~f the .i'L.ft...i';.at~.~daE~~.~ ~SL~`aE;.`.~:~~'.~t~ c~a~t.: t?
fie3 3T24th?'lun;u 37l#z.c i=4as. injected kntmi%e#3E?i.#.sly,
i:F'#yi?cFiTd1`i:FllA distal. W ths;;=t.ct.lus#.{3tih<:C3
~w fi4c, -ati ld low o'3' S>4Ã1~l ~3:~3.) the F~~`ellY~?or~"'~iS~G,E~.
j3i~.F"Ã of the .F;.Fv
z?~#iF
~~ `~.':a{. : :xe NI~~ i;~t~~g~'(F'i~?tiai ~.; ;~~rttu~':~.~:
4t~.~~.~. ~r~~
4 ~~
~ IN a3.,#~f~
Ext:tliopatiu bile duct is oftepi ditBui.It to identify .~~~~hosit conÃra&
agents in r:.~>:.-s.
20 '17hd: 'tise of N fr:ieÃhyle-Fle bz4io i'Ii1:S 4,v#?it.#:3~.:ed :#F r1h
desCr3bQd in Yi21S E4aaiplQ. The imaging St'~toI3i dCtiCrI~.?ed in Example
<.,' was used :z~ thF~,s:'
per#.mC.#3ts as AdfAhr?tf'tre B:?ui"
IrI tLil.e s tl3~~i z e d r aaI ". 10 0 ~ ii .`kg t,f lM B ( l - r#rg'k k;) w
a s i :n;o c 4c;d in a..v c?rFc) v~'l v
I5 via peall4e v6F';. .c`5. slii3:altaneo iss color 4'ideo:`NIzw
fli:#.o.F'e",i,eI:ce A3:itF`a<7pez"at: YL l.t'aa ig1i.Fg
~~~~em v'ws. ci?"a,3Ja#yed f4T# quaF1t.F~'#.5..~tIa n of flt1c+rGsGe1iF t.
images. For 1tEaftW wc1v64'tAtrffi
0.~:~5ml of I Ã~~LA~ ICG (75 nmol:1~) was i.qjo4ted. i,it;.av;<r oi~~ly in
s3aC~3 hepac x'teT#i34-rcipi;.y and pi,rti3 ~.1 3.ppd' ay t <aF7. bi'= sec#l.
111 c3:oA4.etized pigs, bilcCiiJC`=E was vFsuali.ti;F:'3 by 1MiÃ.'%FvL'aS?%Ih
t~j <'c(t(xn of
30 3 00;.#:.l~rkg of 1zi4 M.B, or ta.iÃ:ct +A:3:#aI3lY.~`F,.z''Ãi[?z? l_I-Itei
ti.S;: oaF.13llo-n b'iG;'. s:t3ct f;3x a 31ectiC1I'¾ ova`:
iO ;.M of NIB, Fii.it3T+.~icenCo Fa3teasft:e4 of bile taFli:=[, IFVer (F'Igj3t
3i?hsv), and ...:'3iom3nix,
weie nioas~`t.~~d at.>. 5, 10,f 20,an4.S 30 miFautesfoitow#atg Fn.jC:6.t#on, F
orl':ep:sfiiNc3>
` vtcI'.i~:..,.~ ~ ~ ~ta=3~1~% e T i^ 'S t'3/~t' (`~ ICG ~ ~ ~7~ ~~ is a '1=
c` s~ ri n ~4~E3~ ~:;C?#'~t~3#~~'c~~3n.a~4'. ~.~,~!~7~~,~~,~, i~fi?l?~ ~T ~?
r4C:~F:~- a~1z,'av..l.
zo"l7SiZy.
L3:
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
I n rats, flue common bile duct (CB.[~) was, cl~~ee Ãab lc;just 2-3 1.~i-nutos
after
MkÃw~~enoLis aÃ~~jecLLon ~).1 the MR Tlier<ÃÃio o.Eic~fm-moi.! bile duct
Ãant4ns:Ãty ae1.a~i've t{l
p<3iaZ:Ã"o`_';as and L.ii l:voT' rapidly r("s:chod mi3} 1iIAi:Il at around 10
minutes and wass4-L43dv' at
lea-ft 30 :i1inuj-t..s after Ãrajec:ioia (Figure 3A). No dyc s;-aiakfiq~f #.i
t~:~ liver was ,~?~_~~.. ,~:~.;.
y
The C13D coalai be detected 4'3ntv in the t
llaoI'o`E4Ãli e tl?ia , gc and .ÃI.L qgQd.1>i1 a rri , ht we'i`ei'.
n4`~ in thi c}it.eàs lliige (3" ~ gU3 ' 313).
h-i p}~~.'u, b3.lir had a faint ~~-110. Ã ~'= +~ falxv~ , il;F?d con}:tI3o37
br3i L:ui::r rxaS d,. , k,C:tiible as #.Ã i
the rat However, Ãi1c .nL'iabG=x?iÃtic btlc,'. dilCt was not tdowÃ.fit,ble by
i?i7riÃ'?ghepza.zedÃ5soci;.ol'.i. Direct reÃi'C3gi'adi'
iEdÃ"ilii3lstA33t:t?II:.,f MB ..a3to ilic CBI)
sÃxc.~sm l=r vrsutÃ~ized the CBD (Fi-;me 4). 'lliis dzroct iq;e:.tÃon of MB
ÃnÃo bile tluk.;
Z-'m,t=i1cd c40a5tL'i 'L}tit~Ii.Saldial.dZi11.~~~`~.=o52 1.ib.7..F~~ruV than
ldxdÃF~4S{.ii'.~t.Ã~L4b.` Y-~fCii~i.~25:.`ts13l.[t:i~i\ S..
~~~~:Ã~~.>i~~Ã~., if access to. the biliary systc~.Ã~~ii is z,vmlable; dirc:cÃ
.Ãn,eddo.: (1Ã' M B is 'j-11
ajt.ematIZ.. 1-nL`>i,L3d, fÃ?i v-PtEaalÃzasÃwn of the bÃa`i1,"' S"..stikm.
3+fi re~~i'} ~~; t ~ L:.Yai~' ~tr~~+-'~.~t=E
50 ~tL of 100,Al IR-7 8Ãi or CWStltx (.".A (15 ~tp`kg), or ` inM .CG (150
injected in.Ão poS.Lai vein 1ÃF:5.t2.iArs{.tAe.`ii..k+t}i.aÃi3. A A
a`ti[FSli.~[t<]n'b4iLlitcolor vidt,o. TQ3\
w}.6.xorti.=mA=k)Z.=4iSil.dL.CL/At1W.tai)vW)ÃA?agil.2i^ 4't}~stem was employed
tf3J l~t.f:[.1.if,AA~L.='CLf.~~iiw.\,f ].F_t.tZ.
~2{5 fr~.% t:.'1.=a?S.r ~1 2V w5~ ~ ;~c~. {y g 1~ i32.~x1)t2i . a.. rats, ~
S~f ~+ \', bile Fl~ t~( tdVt ~'r'1 { x ~2 ~if~ )5tfE.=l~ b$ ~`( T~ ~ -~'~
w5.4 ~q ~i2~~'~.=iTti.i t6rt.i li2:5 SCA:Z~`~ Ã~~{2{1.G':41.,~,.~:~ ~N .,1.: 1
<,r iectimn (N-16) of 50 pL of CWSOO-t~~A at . 0 20.. 50 arz~.~ Ot) a,M ;;?
20 i.`~11w?:1f~` :hc E?esi et`3ncomiat1oi1 foÃ" :?.1lc dtÃC?tfmag1#3g.
liver (:~ghà ;c;b: )~ and pancreas were mea4urcd at "a, -'S, 1(120, i.i?c~ ~0
min ~ :c?,,~ring
WIthtn i=iÃ'm m:2'xizti.n., afteÃ"pr?i'Ã.cil i'+u13:1 iÃ:e:eCtÃt7:1, all
ffiIz.e
h:-n<Ã.g,ing of the bile ducÃ. Both IC"G <~id IR-';N'6 could be vismaliF:ed in
:he, CBD, bLit Ã:{:ey
:25 s ~.c~ ~i~,Ã~~fiiar~à dye stacking to ~?f :li~J: r, which à i~ t c? bright
Ãlu , reseei? .c t:ia.n sfw
liv,.: (FigiÃ.ro. 5AwI3). CW800-(YA also shc}v,,,ed Ãttc CBD, -wzrh an
eEc;.lIeni ~3Ã~~~.ltÃ.l Ã.o
background -raz=~~ (SBR). but wit~~i no ,igrm'ficant d:yH stackffi~,Y to the
liver (~ ipa~~ 5C).
Txm~; CWSOO-CA was sellekÃed for CBD visualization.
I'a3(' ilE lxt cxnd zTnE:3t4 were 3xa"Ãz:td%fà o
p1iili~ $Y7~ Ollik:' iIose axad admii1.st:i:a37Z'S T.`i?~~e:
3:3 As ~~~~own in Fi tv~s 6A-D. in {~ ? ~.~ec.rax ~.l, ~'~~1~ could be idc
Ãi~#i~.~~ "~zita? utc.s after cit}~e Ã
~ ~ . ,.
:n.T',AvenZ?i:is or "pC??'Ãtil l-njectiC,n; blià portal vein Ã21jecÃ2on
improved tzxÃ~ SBR. .~~ pe.,ially
using 10 and 20 ~tkf CWSOi~-C:`~ ad.nffiistra1zon. Peak, p~~rfim-nance for-
CBD V:~sual.za;.<.?a::
i sp6 aeC:d i:: 's3~?~3~'~?:'~inli.i~'y~i~v' 10 1~1Ã~'SLtÃGS Ã.Ã1 ~~~:i4`
[+,?n~:E'.31t1'<$tÃt5i-i C;fC:WSOO^CA, <3:I:i~ 50 #-:N:
,24
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
injection showed ~~c- best SB:R. S.~.Itbv]00Ã.:~MCW;s00-C`r~ ~ ~ ~
~q.r~(g) adÃnin-ietraiion was iiot always better than tbat by. 50 ~im CW800-CA
Based w> >h'. results obtained Ia `~. rats, CWcZOO i..:'~ ~~siS as a: it,,'~~
<:E;ii... tFkri~y.E. ~,f a*_ Ã~ #-
. . . . . . . . . . . . : . . was chosen ~1~ euz,
~ br a lt:rges:nimal znod;l appro.:chin~,F the size of humÃ~1. In '~
~nes~~ictizeÃ~ ~~igs~, 5 mL or.
100 :Nt CWSOO-CA. 5 ~L?. kg total) was injected ir3.tt? portal N'S>:it: (.'SV
-- 3)? s.3r<` -;::i.Y.v. of 50
&'-.g 'ota.Ãrespectively) wope
:Tice C~a.~~ ,ntr<ii'`oltt3uslv.
As shown in Figure {A} tho:'3oi"7:11<Il CBD could bf:: v1Su.i.i.zeL`"a us.lng,
5 nfl. of
:
;2{. ~ ~:~ ~ ; \`. ~~ ..~"~~: O~.~:~ (i `~ Ã.~~: ~~?;.~ i ~+~~ ~~~,~li both
pi~zt~~l tt-_, .~:~~a ~~ ~ h n:~; '~~~_. ~rw.~~
~ <?zl ~'~L~ ~~~ a~z~~.c.;rc}~ ;~ of ~s'A ~:~ ~t.
10`0 CW800-CA (7,5 arici 15 i.~gikg. respectively) aiid Intra'vex~cn~~
~~~joc~jon; T. <O
rs kt:=~i), Vasualii~= 4i:~i?IIi1ft}-lÃs *. cE~Sa'i ~ ~ :1Evj possible i}"3F}
_.~"?l~~~S: c. ~.~~ia,r'~:~ ~A.~r'i Y ~iL~.;: =
CBD was :t=c ~~+i3 1ir~l+..z..
Lw
Bcad, of 2.5 cir 3,5 mm in ~~im:ct:u i-~`erc irxs';;rt,,d ir:iz} ccTZ~~inon
bile du::=; z'rr:;
p2pildF;;: icvt.r: by d;:odeii.ostL3rn;? (N=.~) to 53Iiuflaie blockage F3a
thi= CBD. As 5ht}wk, iz",
Figure 7B, iv.i.oT: th.o two types of beads weI'i:= itise.eC4dif.:o (~BD,
thC~so beads could b:;
~ietewied coa-iplet~ly in 2 of 2 pigs, wI-iich were admin;sterot~ ~inL of 50
~mki :,`WI8:,0()-CA
mtrai~enou; ly.
d+mazying
' ...,. :,..f~
~~..~ ~`Ii~9 15 t3i.''., low LTii~ S.. A 1?~C:'IR tiii~~~., t3~.'sii`x~:
~`.9~. ;'0 .;:#..luõ ci.`w SC.~ is :ils-,o
2{y !dec3.~ :fi9:`;.3:ae F.:1 Ã.+3'i:',biI..4t,i`it3I1 :i="itlINli `x91i1
fluorophores i~q?icii~ly used ioB.' guI.fll'fi,.t_ C:'^.'#.iC.h
~
:~} IC;3;~ 1... kMo--channc=i Ns_R fluorwsce<nt imaging method:4. Therefore,
MB was used to
CBD F used .
+'JS~.~.wl~~ze \_-B$ a~~>'~.n i~.~~~i.tirhGt~::4=~F ~~ ~w, while I~:'lF
'4~&~.1ya~:~f:.~i.f to ~'i :~'~.~3l~~~i ~'t~a~~s<::i:ture.
:31`s. aesial"3,. shown hl FigEue .'C, dt;ll7oifat3ate that a
F:.<)Ii'iblnat;oIl o.t MB c1atha
z~tF`kf.'i'w.`avoicõ1Lthi cinl$tln~,~ l yC% such as ICG is ffeL'ttvtr f4)i'
eaiu;s:l%'4 c3'. :ii'.nf,'..3.h .in1c.#c;it t~
~'..~
'stFlZ`wi>``s;` siYiui.itt n~,'ots ideniif.cisE#t3t:t of 7Iiu3tiptc tissue
ski"LE3::tCro4.
~~>.~C 8d~:3i:=~~
Direct tkiC:ctioI1 of bii#.i: dyes into tkSsueh ~.`,a3i iT;dzScri zninci.t#'ry
i:harliy.' E~i~w C'.-i?l3;' of
tlssEaes in ftwsuqgrc~~ field, 34`h#cli cu..i.-, lead to dif"ficu;:y in
dissecting t`9s",3t IF"ziwetif:fn of
blue dy(: Za.1ro!:%~ly the coIii9 il:i?il bile dl.lct #:i i:d3suf:.k eIat as
th5..'. chF:iagi: o`~ the bile c??<sr
3, not d.otecuabl:f ti`, the }iuman i:`s''c. I-li3wE:vei:, in our study, C:B.D
could ~c i:? ~ m.:tied, w:ta?
:ti`IR in -a~l rats and pigs, boeause NÃR f`uoroscerFce fri3m Ãahe5w z~g&nts
withiix th::.
. 4. . ?..` . .. .. . .
CBD pcneà n.te: O$:tis eIy de:,~lyÃhrz3~gli: fl;ic ti;;aue=, ulthz?t zÃ:f.~-
XIM c.iinn~:~~ bo, dQtoxi.eci: if
the ovu1)ing ":ssvw is toc# thick f'oa ti-zc fluor~sunco to ~encirate, Ir,
addition,
.yy
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
at¾;ofir,ortiseen-L:e in the aange of NIR. fyc}r{"f the tf:~sue ~n t}i,.
su:4ical. f..:~d %va, qua ~
`a'-hu15 thi Ing prc?cedure ::?r CBI) is IfÃgbl} sensitive aiid
Th.t"L`e NIR contrast i?gtvS.1i5 w4=r#.'. <.=i3?Ilpi:fl'Ã:d a6r 1.E3+`.'. in
bile df.a.`c.=t :fi.f`s.ig3ntj. WGS;
k. W.
f"W800-CA, and I_R-786. CW800-CA proved to be ~ie most selective, a4 ffiL.
~Ah{ r Ã:,,vo
co-n~xasà agonts suatcsect tAc+an lack of efficae-rit -qccr<:tion i:,r.o, bilc
~?~o~~~ the bver. The ;?;,.:}:
two agoms h~avo maa=kedl4r:laydrophobac properties, w?.z3 high affinitlt
the cell .?mnnbmne. CWSt3O .1?a4 4 -,.-Li1.phonated rcaii~ucss ~ut .~ivre is
zero in IIZ786 and
m ay I'Nvo su.is no.?aied ar:=siducs fii ICG; t1-ic nai~aber of a~lphonated
tosxdu:o's, is reI~~~cd to ,1-'.c
hydr.?phil;c properiy4?f the contrast agents. In addtf~on, CW8()0~CA was
s:tc.}wn to bÃ:
excreted into t1?o bile i , a. substantially unn?etab.3li.:ed Iba'm, wl;.ich
increases flic napid:ty
wft.i3 <vh.Eeh i-#.?Akg%..s can t?i,: obtained after ad1;slLafStrf3tÃof,,
ht7gges1z.Il;M that f> will be 5aze ti3r
z,5e in hurn&ns.
The dose of CWSOO-CA re-tjta=red f~.~r CBD cleto~~ion i-s in the ~ange oà :*_
< 5 UvI ;-
;C?~ ~3~?~~z V61, ~f?j~'>~t3~3s ti~?~~ '~-i~ ~#,~'~';~g lJC1f
:if'~tt::71;t?~. i.~>.~Y'c35~.:'IE31.ls fF~;~:f:='~fE?.~;.. ~~~A'riu goal :i3
dE`sc i:tctilistn?vnt is io zToF CW800~CA c<?i]Ct'Tltra:f(3n pf3 CBD as clw?Sw
tij 10 #.F.NI as pobsibii'~
ar ~~ ~~i:i in 3as b:ftaiiyTLrlfirt~ as low as ~7C? ifb ~.i. Sfgf1 ~7.i
c3T~t E~ 1Ief~iG~lf,~izg .}x~zflffE?:vscGx2~:-e
occurs Zbove 10 :iM C'WSÃ#O-CA (Ohnishi c:t at. Mol: bnagi-ng., 2005~ 4(31).
should be a..i~;ed that 1: is unlikely that i-l#ght` a,'.~o-n{:ef?~'f"~:r of?.
of CWSOO fnje~:~t3ts1a would
improve the CBD i:;?agzng becau~o, of this fact.
,;;~
~
Exm:?lahoraci. Duct M
- i.? p:
In ~f.~; a:he:ti ze;d, pigs, -S mlr of P"/ ~ met}4~~lerfc blue mis f~~jecUX1
~~-ito a;i
Ivnphnode, then the f1h~~racic dt:~~t was obsen.,ed. Thoracic du.tw~ ~~~~s
visuz.l:zud with a
'`da,i!r -fiai17s blue cfdli:'sl`:i?''e the color imafõe<btti was clearly
viatte`.aFft;i.:Ca in the Mor`w,',"ceTfc?;:
25 i:'.? ;g;:- (Figurc \).
Add]tio-nal L'`?>pv.#"`amG33:5 on fl3i?i'E3.t=1c dl1ct vfS#.~:alF:~:ai.Ã(?AZ
#2: t}2o rat mCd%lv"Verc Ltiw-l
p; rio ~~ed using o:he-r N IR fluoi:~~~~~t m~t~1:dts. 50 p.1v ofa 10 W
solutio`,, cif _CG,
:CG' 1:[.SA (a o$'ICG to 1?.un?an serum. a1.bumt.?), C W8MOCA, .~~:
m ~:lb1':~~'4'31Fc; was injeL=t:::Zi i.if~:o
HSAW.}~~ (a cÃ3'vaaeSet c(?fi~ugatioi3 of MN` AS'110 t> human sL*ru,
3~> meSt.nterfc l;~~p~~ iiode, and vzsmil.arc=d 5 minutes la.:ur. The aesuifts
are shown in z if,.uire 9.
The thoracic duct was 31.+'3. t visible tC? the 31.al~Cd eye. f.3the :;ola#T
maÃ.s,x:.'., bz~:it w{.s f:li: .af'ly
`viSuc#l,::ed fn the flluoms(:<.Ã:ey,"'~ image.
26
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
Ex,Lr,a ~Ã ~' E_}rox~'.It3~:i~~Ã~Tim
In znesihet_.ze~~ 3 pi>s I ni-cpIg ofnic:.fi~~lene blue was in;ected
>nCzavun.3us'iv Feadl
dhaIÃ: tiI~;. abilo.Ã-nan was opened Sti::i;gÃci4Ãxly iÃ31C Zri,'=ter wt3'~
id+,'.n`.'lfii'.G by it'n.~gli`:g< For pel; :,''
i.iztona~~i~~~3~i~
f1 Fk-i E'i2t-; ureters "'ve:re vi3`i.]Mi; ed by intravenous s31~ectICDIs of
5;) +_'r. CWSOO CA
10, 20, 50 and 100 :N l to qii 1nIl$y' the best c:?nE:eFti.$cl~.~:f371 fd:.~:r
bile duct #.naa Ãiig.
?`lu[37`e',~`.f;f,.ns;~ ?n.S,.nsia.1C:` 3i~? ureters, ~i,.S.~.i~v,;, and
cibC~.:.i.~1ini.ll wall ~.3~(:rl'.Ia2`4-`.``~:~..Ã.~ ii~y, ~, ``~
x. r~ ~' > ~
'2.0, and 30 raÃn following ina4=ctiora
As shown in Figures 10-I1, in general, ~~~~las could be 1:.c,n!,.Aied .3 Ã
ain:aE.e, aft:;r
iii'Ã.<#.vL3l'C):l's ii'.ijedid',`n. P;iik perfi31`in:i:nC=e fi?i`urCtE`.r
visualization ,app..=i:ri:Ã3 at
xdp~lio~:r11~,c1t4i~% 1`i~ :3'l*^.s`z`v':~ in C:v~::~' v3:3a (:e:.~~eri3tlt?õ:
oa`CWSOO CA" and ~~~ U.M C~'~`80i.?,.i>:A
~~ M800-CA 05 ; Ãgi1g)
(W ~.M;.kg)i4ec.tiÃ~~~~ owed t,:~~ be~~ SBI.t. SBR for 100 ~~
i3dn13211stiat:fsii w.``~'~.:in5t <sw7.ys better ttIa:i7 that ~~~ 50 P, M
M800-^~.~A i:dnaiia.s::i'iitÃf5n.
~: W~OOCA provided c.xcvl~ent vasualizkLCFn of ttret.ers as, is Sh.ovq.i in
Fir;txie 12,
~ ~amm`r1.c 6: ML, a5>o1i`k?t1 of CW800-CA
As desc.'d e'`beta h'vÃ"e13i, u:G.'.t<:r vEsi?.t3li..F.<$t;.f1Ti can usually
be obt:i.FA`3i,:,d a%q)`L>lv, oL oi: i3i, i~
few 11-~inut~s following rIlii~ suggested that CW800-CA may be oxcreted
1#z"'Ãni`= el1n1?ITc~tei as an E:nClia31ged ft:+i'.F7i, m me:ht.3lÃsatit3bl
would likely &s::o'S''
,.g . , - = i yq~ ~y
~'~~' ~,'.:~i~.~~.~' t~ a~Ei ~~`~i.Ã.t.~ an d-3i`TrC.'mxtl :if)Z;=4t..omLtry
f:i3'i;in4(:=~~.~.~.~CZ,`~+~~
.3:-fliÃ;ht (E-S-'.;'OF) rnets;iod has :?eeÃA s~~~ide;,
tjai3c ~;~>Ãpzes were c{~~~lockd. ftotn pigs (N:-z:5), and xna.y., i:=d. The
dt'Ã`. Ã-Fs17zffly C-....
b~. pinpointed w:theà fluC?rd.::sceTÃL'=~~ de-teL:=ti..~r equipped wÃtll Es-
rrOF ;'3.ass'
U`i'ÃF?esi#.Ã'i:plus, wom,' fir`t pas5al over a &000 D.,~ cEÃtoff 4el-
l"Htz:Ãt:trn oC3li:imÃ, to remos''~
~?r+~~~0'>l~s=; t3,~1~. ~:lt~2it~'.. less t~'R<1.I7. C3.,~~~~~`.~ _~:::1. was
i3l2:li~;Z(:~o ~.?I~ i3. Z_` cc3`~"3tÃ:
A3uffor A was wate.r, Bi<3F1us (Arnc Ã'ican B.~.~a.nalyficaa. Natick, MA) and
Bufffe: B~ was
Ãi~. Using <i. gnadisP;nt iif 0 tb-.s 50% ~~~for 'B ot~~~ '0 mir,, urine
s4inqp1~s were
i'e:4iki ti'ed 3.n, a 4,6 x 150 mm vey'Ã'.;~xi11etÃ"y C 18 column (~'~r
i3t^wi4) z$l a ilow rate f i~~ i ili: i:3er
minute with e~ ua',e fed iiiif3 :~ Wate:N LCT ~szi'oi`ma:~~ spe;Jt:-on:s;ter:
Ron rna.de wa,"~" sw:i
'1'v`s w^'vrC. si:t at iu?d 3 000
to 'L'.l b.trt3s;p.'ay' negative (ES ), E3rfd cone :Ãild i :qTPllzl `4''
4'i3lta
(V), ;t:spect:vel y. D::sf?ivation aiid Sotircc ien:ipurature were :.et at
'50S 140 (C):
;-cspew~-tively. DMa were amilyr s:d. wiEILi MassIynA (Waters) software,.
21
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
RLsF#hs r#3e ::how'n in F#4?;2ik..'s 13 and 14. f hS' flu{?rE'.sL.fi~'?"ici'
fhZ#n the :?r#~:53 `cimp`s.'
was ~~cte:.red aiid pinpointed at tlic identzcal mc~lectflaa weÃ~T~~ t of
C~~~~~~-CA. which
su<3<~~s4~i~ that CWSOO-CA t3vaÃS excreted into bFi 3c{ as ' z ~ :vi`'w
3a_fFtil:C:i :~i7I1~F. 1z3b= < a Ã.1.;~.<:~~ ~i
kse:~ t. an ..Ã~31`i~~. 3s
i'.o:15.ÃSÃ-Ã'71t w8^.h our tex"i.?Z'.a,:ta:b:i7-n.. This rapid chi-
nix3.3tTf2:F1 ffi?Ãl'1 Ã.i3e btif~Zy'suggi'.oted
CW800-CA ~hf3uld bt a %ate i;g.;iat for use in hum;atiis.
~: ~c~i~a a { Reion4tri#ik]ve Suj~.ti~ry
S as.Ã~..f=.co.33 i PC~~foF.mi:ng", e(:'S)nstru(:=tà w`L.. .~~.'F=:~~ycnes
;,..eods a s`aiexsÃ#a;ple, m."`.mm'w:lly
invasive means of 3m;.Ãg111L,' .3Ã?i` di:l'4nn1mI.Ã?g th ?ktlcl~i??Ãtf tÃ~
paEff:.n {?: :..3trsi key
?i? pea ~s`#'"',.iaa,; vessels ,:Ãi. r::al-t#.Ã~~it in. the operat.Ãng
Ã't?ot3i during the ~ivnzng and oxvww..Vn of
fl%:pi.5'uÃ'getar'. In i#dldEtI.C)n, IÃ;.:he pÃ:Fsl(,q-i#,'.Ail.fiv`r'
p=L:S"Ãt}da ttPo cl3_nf~~i'm ;icoil4 a ~11-n1~~iia?' F11eians
fsf._Ãssessi~g flap. 'vi.ibhlÃtiy and ~.~tddir#.~ surcsica1, efforts at flap
salvage whon i .;?mpxic-ali,<n.'s.
occar.
Al t.~it present ti.no, tlicrc is little if a~~~~ intm:iperaÃt?vo iiiiagi#3g
p;;rftmnid during
e"e,",.f?#.`hL.~,~,m#-vi:: s.8: g~i=i~?. The z'. t ~~?t)~dti fE ~T this
~311..E1 ~-3lf;.. 1.-' ~.5:, ~ ~ .~5 f.~% 3 ..:`E:t~~.~.. ~ s.. s are ?. ~f
z~'~.~, Ãj:ci3. as
i:i3mpw,c;d eJrni3gf'<Ã:?h'v (CT), T1'#.qgi.eft TesiJfic:33'aCi;:
]1"i'3a:;f7ig (F 4r~RI); single photon Lm#.SsÃ:_i'#
:.omp?.it;d ti,2'I3Ã?,.i'aphZ+ (SPECT) a?nd, positron 01 11s5ion,
i.oFni`,g,..~'iapby (:SEI) are ;nÃpnaCi.ica::
i311t3.'~C'so,~~Si~y :':`,3' intTaC~pGra#.?v`~-` use. 131t3'aSi'?uÃ4d
d5pf?rtdbto; a3 nC2p1'ob<:s a"FlS7 be %t.i.dE'?`"+:fa
. ~
::t~ i?.5:, ~~~i3~~3~t'~'C:``, it a ~1.Ii;i::,..''s L'f3`ltiact +,=~".~1 the
tlssl'Ãf% ~e.tii~{~~.1 StS~.C~y. ~7_F~:s i#.`F~.1.k:.=f..
....~:ii";.~1.3:~t~3`; >:
20 Il#. ~3#ter ; .,, tf~ '" v`+~. ~ ~, : ` ~ = i~L~~.~). <Ã.~aCa =~ ~ has
1:dT1F'~4.'ia ~ options 'r~='itx3 ~ 3"(i:~ ~.?Ci:t ii: q ~.t)aÃ+:~'e.#:2 s
~.~i.=; (F :=nC.~"i1t '~F~.)~- :
.. = ~ ~ t .i..=~i: ~,
`c~.~I#;'= only fithCGs modality avi7.l.lbfi, int7':3p+,.'I"r#.tF4'tly
x'o'I.' 14.=#.`C'.?n^itlÃ.1HfilC. s'3.Erk,:-y i\ x-ray
alllw#t ~i'~~3~,`r'. i~f'~'ti'P"~ r~Ã~~?E?gÃa}~~~~f 3~'o~.~f7$Ã'i:=`,~'s3
dedicated i::`iC~ c7xe?,Tf'I?Stt`t~ F'~t~ ~,?;~.Ã'L~F ~?:a~~'y
3'Z'ql~#ri h exposure of patient and caregivers to x-rays, and exposes fl34
pi.#t:.Ciat lf? #.i1lil:~33ae:`z;;'
wnwentraiionsof nC:~~-LrliÃ`ox'iciot3.Ãm. .Empoi;tana for tIl,is iiLudT0+,
angiography wclgh =:? all
ne, vessels ;~~~~~~~~~~i-ng a ;~lap cyLmlly, itlthun.gbin casus of
PC.rtb:ratc}r artery Ãdi.=~ififi~:atio.>, vbw
-' 4ssezs t1iF?ÃC: wiri7n'ly.
~dec~ I;i3i? Ã IEa;ti tqi3.iv L~'ou.ld w?.gi3t surface
`~ e, 4vi~luatethc: usetu~~~ ~s o: ~lR4igents i~~~ ~uchmFuthod4, 0.5 mg(14
u;.. Lg{
3ndt`+f<3'aÃ="z1 `#ti, g>e#.'.kl (IC~ ) w'as ?.i3ject[NI .Ãn~'t#'aN. S
.Ã7;Ã3usl'F' :Ã;.lti? :a 35 kg YciÃ.k5h'Ãre pig, cÃn'd l'ze
~~~~i w:~.., image<a as tac,sct'ibkd herc:Ã"it.
30 The results aÃ'e Sh(?wr: ÃI2. A'IG. 15. Shown are ;Z.ew,'r1jS.G=I7:3?i
3ut<AI?:iG3"'4;4i:i: nci; ;:topi,
;.~.Ã-wrÃ.i-~l filling at 45 see ~`.o:^'.t-1I1;'L.''9::~io.`.3 (second row) wd
venous fill1"?g at 10 ~ t poS::N
ir~tccfdcma. UsLnM, . thi::~t'= data, the exact li7CsltÃC)Il of each
pfiriC?i'.;EoF' is ,Tlsi?'!ied Z?,.1 #:iõc. <kÃ:.e
with black =T3a.f=koa'r` and the flap is `x=le1';.itel`~ (b4PEtorn T4.Z4'' ).
NaR fli?.i,;rLSt;C--'=n..+. ,#irt3<3.i.T,es havw?
12 S
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
#elon #ca-11 expoyurE' times (67 11ikhec.) and II:ClIi%13liz.#t4l>i7s. Cllie
i.T1a4}i.s were :CquÃfL=d c':='G.ry
200 ÃÃÃ5ec, TiNlEste rlitiÃt the nil?ple in t>?c field (top riojit) serves as
<in :Ã.ddzÃiondl intom.Ãa
Co#3E.p,'ol -ft) i#1te-rA41. vs, venous phases. HE',iiec, in mal-t#T#ie, zuud
w1tholf.ti .::i~ui-Izlg
lC?n.AÃ11ym6:#.t.$oFey ;hfi pt'T"x-oI ;iiiAl` r iteI#e,'.~ i3S:ita -ve1n4 can
be :dmtIfi'Ld `s.si#~g N.IaZ
ahi,,,E LsLtits indicate th3t the u4v Clf NIR agi'3:ts, x'Sid U}lf#,fz f.aM1,
methods described heseÃÃi, are useful R3Ã flap design, assessnÃent f fl ,Ã.p v
Ã.b-ility, atÃd
sc~cctioÃÃ of 111ÃtiÃÃw; flaps -ftir salvage Ãho-rapy ,A= #th
~`ibÃ.`Ãnoaytacs.
vt iÃi_WE3~~engÃh FIu.aresc.:n;e
1Ã, To f'~r`~4#:lE#.3l.ti1d :3C3?.}#bÃl7:1 '(' of 3.11:E:xlt# wa6t'i:lG?lgti#
fluorescent F nnzzg7:ng W:_. . ..i.~~~s
.rn: thvaL31L blEÃ:t,:wi.'Ã"e used.
rhe :'Gs3hs <7iv- "Ia(?wÃ: in F`#t?vii4 16. 1 }.2?g"kg m#.=th}`le# !L: ba#.ic
TV iÃEji:i.E3.=sl, wah
iollE?wt=d, 30 mIa3 tfltvT by ~},~~ Ã~1~'~Gg I~~~Xl IV 1i1j+:i:~.lCi:Ea, As
can ~~, s~ .~P`: "1:,.: ~.~:I ~s. >(`.~~4' E??
in Figure 1,65. the. orientation of ti1S C.ia#I'e$#lt?n bl:lE` duct (CBD)T
gwzlbiiodfseI. ÃiE?.fc:k
(G ~`~~), wid i:ystic. zÃ~;-teÃy (CYA) is welI -0swahzed. NIR caÃÃwra
e;~~~~~surctim.Q was 100
'll'ÃsOC.
NL.=:{t onnC% of fiha. d1aksoEii<.kl branches of the i;E?zi`11i3i'!!'
ans,+r`y' (M) waN E.3wmpvd :NWhlEe
&rÃfsw3,. and Ã;hcÃ: I nnn/kg of`rÃÃeth~,~lon4= blue was ii~j e~.M.tect
inÃsatvenÃ:fu5ly4 4c: One second
mw of Ã##3szgt': j3#stiAftE# .ÃewE1sI17g:he c3c#IYl'#3 0,03 ll3.if'~~ ICG was
#njvETtid #;??taava#aL'>#:sl5.
20 D.' vv<l:a d i.`.Ei'.L:E:E:d clearly (;.'i'.lli.3w arrow). The
ri',t~F`[Yos# image in. fl.-w sk:c+`.`+i.i: iwx was
created by #i1orr?:i~~ the i:mi:lg~.~ of the coronary aT¾Ca'1ogii3phy
E#.sf.#ig .ÃI`c1h~1L,nc ~.?13:#t. at
chur, pi-na D2 aaid t?:~e image ÃÃsing I~ G after declamping D'~..
After iakÃ,E, the 1.CG was evenly distributed over ffilC: E:min
:?'#4'oc i`s'(o.lE##i~ ~se = third row of Ã:n's3 Y=sN tlIC red pyet1dC?Loli3Ã
the mer ~ed # > .ag~.'= of 'C:.ahi:E.=Ãc:E:
p^".xfl35ioÃI;, however, a pf;I`:'EiSio2t defect cvE`sld still be detected in
7 00 ÃÃÃi1 fEltllG
;.-lak3.Qseence ÃÃ-alage. (gÃ`ceÃÃ ps,:odoCo3or) and in ilic Ã'i-ieÃgCl image.
11.7bk. area w}::Ãc bÃ?:h,
i34:y c3.Tf: :~is''LIibi]ti'.d was a wt'%k: t);'zu#gE' pseudocolor.
15 >:T}#.i3:i,.te4 afti.~i' 1CO 511;ji'=c`s1JI'!, the :ieaft, was resected and
."ATF#nei:i3;tE.ly vxwr,`#?ue.,;
~i~i~ IL'=~Ia~~~ .e i: a~1C2~~'.Ã~ zY2 ~ki Ãi~.1`: gf'.6 #Il -d:w}?C:+ftibm
row. 3L> :`,t327fir.: ~l+. c{. N-Ir, E:arne_t=.3 exposure 'Ui.ÃaTe was
3`ziSee,
,9
CA 02692923 2010-01-04
WO 2008/076467 PCT/US2007/072800
Ex~~~~;n3w ~~:Parasl;~rorc~ G1a ~~~~ r.~~~.WJsI&' ""Sir~(" Njcthylet ca~~lue
(1\4~)
T(,, d~~~~1-1irla.r wbothe. ~~B fluorescence emissiom could be iasM to
~rnag:~ie
para1h~ro~~ ~lar,d.2 i-f-,g'kg :~~~B,,~-=as ad-miniyteroci over 1.5 minutes;
along witti
p,rif)r :^u~w=Q:Y3t. As cm be see-i-k in Fi~ire 171 ihC+ pc3raLhyrL22(a was
i.ndwcd vAsib3.t`.
OT 8 3.~L.{i~t EMBODE.M.8.'.. Nf i=S
IE is to b~~, understood tha} while the 1nveat:t.on has bt.i'.n rc+'eticr, bvd
33i Q-C~:+zlÃl.a.actli'Ã':t
,v.th the dc:ata.ilod dosc~i.ptiz?D UICa~.~f the fbxC&Wlty- dcscrip:i.ort _is
intended to iifiust_z:~te and
not lIn"It the Sc~ipe of t}3L 'ii`evi:ntl'.+n, whff:h 1S de<IiiE>=d by the
scope of the appended t.iutMs,
Otl3'Lvr i3..to-pS.c`.S xidS':antc3.+,es, i323d modifications a1`'e w4t-h1n
txa4< ,.cL7pe o. t-he -fLdk?w#~g ch3i`.'Im
C~ t3