Note: Descriptions are shown in the official language in which they were submitted.
SENT ~r:L~wE PRIC~ L~LANC ... i 4- 2-9U ilO:4~AM; 703~841124-' 3~1 13~8;~ 3
~ 01377~
~H@ WE B~ L~ 011 ~HC ~IL~UY 0~ PW~lC
1nv~ntlon l~ d1r6ct~d to ~thod~ f~r troat1ng
wound~, OU~5, ~nd ~br~lan~ d ~ ln~ury, tr~uma, ~ur~ry or
bt~o~ th~ou~h th~ d~llv~ry ol' thar-p~ltlc h~-l1n~ a~t- t~
uch ~1t~ls. ~ p~rtlcul~ h~ nt1~ dlr ~t~d to th~
dellvory o~ th~r~pwttc ~nt~ ~o ~oundt, cut~ ~nd abr~10n- on
th~ corne~ or oc~ r ~ur~co o~ tho ~y~ 11PD~Om~ and
11poson~-~ol~r c~pwlt10n~ ~hlch ~wclflc~llg ~dt~r~ or a~h~r~ :
to tha ~ount, cut o~ ~br~lon.
L1po~ r~ uc or v~tcls~ yor st~uc~4ra-
m~d~ ol~ pho~phol1p1d~. For ~n ln~d-pth ~l~cuJslon ot th~
pr~p~r~t~on ~nd proportl~ o- llpo~ , lncorpor~tlon o~ d~ug~
Jlnd p~ot~ln~ th-r~ n~ t~r~a~ ~ru~ dol1nry;
~r~tr1~d~ D=~x~ Vot~ nd ~II, CR~Pr~
~n~.~ eOc~ R~on, ~lor1~ ~, th~ cont-nt~ o~ ~b1eh ~r-
~ncorporJlt~t h~a1n by r~f~r-nc~.
~ h~ U~- o~ 11po~ d~l1v~ry vQhtcl~ to~ dru~ or
oth~r ~h~r~p~ut1c ~pnt~ ~ kno~. Llpo~ u tol1v-ry
vqh1cl~s o- th~npwtle 4l!~nt- to th~ ~ra 1~ ~1JO lulo~n t for
tx~ , U.S. P~t~nt No. ~,5~,0~71s~ d to R. K~ tan, M~rch 10.
198~ d~sGlo~l~g th~l tr~-tm~llt o~ tr~ tte or ~ 1e~1 :
ph~e~n~phyl~etle ~ntophth~ ltl~ or l~V~ltl~ o~eurrln~l throu~hou~ :th~ a~ ~lob~ ~ toP1eal ~1n1~tr~t10n o~ cyelo~por1n tl~
~u1t~bl~ ~x~lpl~nt~, ~ncluglng ll~w~ . U.5. ~a~nt No.
4,6~,57~ u~d to Fou~lttl~ ~t ~ y 13, ~9~B ~1JC1OU~ th~
pr~p~r~t10n o~ b10-ctlY- ~g~nt ~rry1n~ l~P~ 1cl-- ~h1ch o~n
b~ ~pp11~d toplc~lly hr th- trut~ant of oe~ P d~ a such
:
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S~NT BY;L~W~PRIC~ L~B~ANC . . .; 4- 2~ 47AM; 7~36~41124-~ 3~1 139~;~ 4
20~ 377~ `
91~uco~. ~uto t ~ 1n ~C~
Vol.103, No.~, M~ 9a~ ~tclo~ th~ d~l1vor~ o~ 5-~luoro
ur~c~l ln ~11p~ o vl- ~ukon~unct1v-7 ln~-ct101l u ~ ~thoC
for lnhlb1tlna ~l~roblu~ proll~r6t~0n ~ollo~ln~ e~rlor
~cl~r~cto~10~. ~kut~ ~t ~1. char~ct~rlz~ thc~r d~llv~lry Iy~t~
~ ~n ~d~nt~Dus, ~u~t~n~d r~ s~ d 11v-r,y ~ t~ prlon
~tho~s 0~ tra~tm~nt ~lth th~ ~r~ dru~ r~qutr~t ~Mquult
~ukon~nctlY~ etlolls.
-
Th~ u~ o~ t1po~ or c~rt~ln t~p-s o~ t-r~tet dru~
d~lt n ry 1n ~h~ hu g~en d--cr1b d ~r~Y1~u~ly~ Horlsy ~t
~l., ln J. I~ ulal " Vol.~6, No. 2 (J~nuary 1~ ) d~clo~
th~ t~r~ottn~ of dru~lo~dt~ 11po~o~ s to W-rp~ S1apl~x vlru~
~SV) 1nt~ct~ corn~l c~ , 1n Y1tro. Th~ t~rgot1ng ~o1~t~
1ncorpor~tcd ln th~ llpo~o~ ntlvlr~l ~4noclon~1 ~ntlbody
to ~eoprot~1n 0 ot HS~ cont~1n1ng ~ en ~lcntly~ounJ ~tt~ ~c~l
ch~ln. ~hl~ ~1m~4nollp0~0~0~ sho~n to blnd ~poc1~1s~ t~
~SV lnt~ct~d r~ t corn ~l c~ . ln ~noth~r ~x-~pl~ o~ t~rptcd
~ruo d~11 n ry, lntr~vltr~lly 1n~oct~d ll~o~om~ r~ uud ~or
dc ry of drugJ to tbo e~ of th~ r~t1n~ m~r~n~, ~n,
St~r~ v~tl~!t~v~ ~htl~ nd ~ SC~
Vol. U, No.~, ~M~y, 194~). U.S. ~tont No. ~,7~6,~S~ 1J~U~J to
H~th ot ~1. (Jul~ ~, 198~ tl~clo~ tho u~ ot 11po-o~
~nc~p~ul~t~d ~nt~ ~eh ~- ~ fluoropyrl~dtn~- us- hl 1n
pr~Y-nttng ~c~r ~on~t10n b~ lnhlb1tlng c-ll contr-ctlon ~nd c~ll
~ro~h 1n 1~4~tH~ t1~w~ ~n th~ ~y~. H~th t ~ erlh ~
~rl~y of ~onl-nt ~d1~1c~tlon~ o~ th- ll p~o~o u-1ng llg~nds
~nd ronptor~ ~or t-roottng to p~el~lc c~ rf~eo~ or or
n~ blndlng.
Th~ pr~p-r-tlon ~n~ u~ o~ b10~dh-~1vo 11POSO~J ~cr
eOplc~l ~ppl1c~t10n o~ thor~poutt~ nt~ to th- o w1~r ur~ac~
h~ o bon do~srltod. ~oslt1v~1y-ch~rg-~ llposo~ nd
po~y~4r~ ~r~ kno~n h r th~1r ~on~p~ct~lc b10~h-~10n to ~urf~oo
:- i ,. ~. . . . ~ , ,. ,,,
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SENr BY:L~WE PR1CE L~LANC .. ; 4- 2-9~ ilO:4~AM; 703~841124' 3~1 13~ 5
2~1377
rl~o10n~ o~ y~ , F1t~r~
~b~l~, Vol.~9, 1~p.611~49û, ~ 7)~ uld Shlot~ ~t
AR~10 Annllal M~t1n~ Ab~trut ls~u~ p.~ O.~a ~ 3.
~ ho ~ol~ ~lgur~ lllu-tr~t~ o~ptlon of llpo~ of
th~ pr~ont 1nv~ntlon to corn~ oun~ v~-u- th~lr rol~ttv~ ~
non~orpt1~n to unwoun~ corn~a~.
~L~
~ ht~ lnY~nt1Dn tt d1rut~ to ~tho~t ~or tr~tln~
~ou~d~ ~ th~ d~l1v-r~ of ~h~r~psqt~e ~o~ ln 11po~ to th~
~un~ ~lt~ ~he 11po~ ro~ront1~11y ~d~orb ~o ~oun~ l1t~
~n~ not to h~althy tl~u~ sllrroundln~ tho ~lto~.
~ or- p~rtleul~rly, the 1nnn~10n 1s dlroeto~ to tho
~11v~ry o~ th~r~p~utle ~gent~ ~n 11po~ o ItouRd- on th~
ulrf~e~ o~ th~ ueh u thn oorn~ o~un¢t1v~ ~nd ~el~r~.
~ea4-- th- l~poso~ cl~rrl~r~ ~10~ t ~xelu~1v~1~ to th~
Hound ~1t~ on thc ocular ~rt~e~, th~ ~ount ot th~r~pout1c ~e~nt
norm~lly ~n1-t r-d ~ g~nt c~n h ~10n1~ tly
r2duc~d. Con~u~ntly, th~ th~i-p-ut~c ln~ox or ~r~tn of ~a~ty
o~ ~ drug d-llv-r~t ucordln~ to th~ ~thod o~ el~1~ 1nv~ntlon
wtll b~ 1~prov~.
AlthQqgh toplc~l oph~h-l~1c ~p~t1C~tlO~ of ~
cont~lnlng ltpo~ h~ pr10r Joll~y ~thod~ h~vo
b~oll u~t ~ n~ ~or ~0pro~1nQ th~ tr~n~port o~ th0r~psut1c
~nt~ lnto ~nt ~cro~ t1c~ b~rrhr~ of th~ ~yo~ ~uch ~ th~
co~n~ on3l;n~t1v- uld ~c~or~ ot~b~- ~nd ~10~h~ 6
llposom~t i~ch u tho~ elo~ ~y Norl~y ~t ~ nd
~ 3
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S~NT BY;L~WE PRIC~ L~BLANG . . .; 4- 2-80 ;1~:48AM; 7~3~B41124 ' 3~1 13~8;# ~
~" 2~37~
.
~hlot~ ~t ;~ tlst7) h~vo h~n ~or~ul-t-t through ~1thor cov-l-nt
modl~le~t~on o~ th~ pho-photlp~d or lntorc~l-t1On o~ nt
~410tul~ lnto tho 11po~o~o b11~ r lho uu o~ co~plo~
r~a ptor~r~co~nltlon ll~nd~, or po~1tlvol~ eh~rpd eo~pon~t~
h~v~ hon d~v~d ~ t~rg~t1n~ ~pont~ or non~p c1~10 ~10~dh ~1v-
co~pon~nt~ for t1~wo ur~co~ Ho~ovor, prlor ~thod- h~n not
r~e~n~toJ th~t llpo~oo-- ~1tl pr~crcnt~lly blo~or~, ~lth
hlgh dc~rc o- ~p~c1~1c1t~. to cor~ool ~ount~ ch U cut~ ~nd
~br~lon~ ~h~ pronnt lnYontlon 1~ h~d on tho pr1~1ng
d1~co~ry th~t l~po-on~ re to ~ou~d ~1to~ but not ~o th.
~urroundlnp h--lthy t1~w. A~ u-~d bor~ln th~ to n ~und~
lneludo~ tut~l ~er~po~, ~brulon~ or t1~ d~ ~ e~u~ y
1n~ur~, tr~ urg~ry or d1~u~. Th~, thc p u~1c ~o~t~
~h1ch ~r~ kno~n to h offoct1vo ln th~ tr~t~nt o- ~04nd~ eu~,
~eeord1n~ to th1- 1nv~ntlon, b tollv r ~ ~pOcl~C~ to nund
~t~ ~1th n~ nonto~1e pho~phollp1t/ehol- t-rol t1puouo. tn
p~rtle~ler, th~ llpo~oo~ eont~ln~ng thcr p~tle ~nt~ ~r- ~ :
u~o-ul ~or tr ~tlq~ ~ound~ o~ tho on- p~r~1c~1~rl~ tho corn ~.
,
~ lpo~o~ - ~hlch c~n ~o u~ d ~ecord1n~ to th1- lnv~ntlon
1nclu~c ~ny 11po-o~ ~hlch ~lood~or~ to o ~o~n~ ~1te~
p~rt~cul-rl~ o corno-~ ~ound ~lto, ~nd ~ho~t no toxlc,
1nn ~ tor~ or ~t1h~ 9 ~ n~et~. Tho~ llpo-o~
co~pr1-- ~ 11pld nJ ~llt typ1e-11y eo~prl u 50~00 ~ol~ o~ on~ or
~or~ ~nthotle or n-tur~lly occurr1nB pho~phollp1d~ ~n~ 10 ~0 ; ~ :
~olor perc nt (~ol#) ehol u t~rol. ~ho~phol1p1d~ ~b~eh c~n b~
u-~d ln f~rrul~tl~ 11po~o~ or ~c cccor~tng to t~ ~r~nt
1nwnt10n lncludo~ pho~ph~t1dylchol1no, pho~ph~tldyl~lyc rol,
pho~ph~tldyloth nol~1no or pho~ph~tl ~l ur1no ~lth dl~c~t ch~lns
o~ typ~c~ 5~ to 1~ c~rbn~ ot ~lthor tho ~tu~t~d or ~onooM
v~r1~. Otiher llp1d~ cl~d1ng pho~phol1pl~ bl¢h c-n b~
u~d ln th~ ~or~ul~tlon o~ tho 11po~o~ elu~e, ht ~rc not
ii~
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Stl`~T ~1';L~WL PRl~E L~BLANC . . . i 4- 2-~D il~;4~AM i 7~30B41124~' 3~1 13~B;~ 7
~_ ~ %~13'7~0
11m1t~ to~~pho~ph-t1dy11no-1tol, ~ph1n~oly~ , pho~ph~t~tlc
~cld, cer~ro-1t--, c~rtlollpln ~nd ly~opha~phol1p1d~.
~ 11po~o~ c~n h prep-r d ~y ~ nu~b~r o~ ~ethod~,
1nclud1ng but ~ot llm1ted to, proeodur~ r pr~p~rlng
~ulS11~ r ~ un11~n~ r (SUY~ r~e un11u~ell-r
~L W J, plurll~m~ r ~5PLV) ~nd r v-r u ~ph U ~ ~v~poratl~n ~tV)
llpo~o~ o, ~. ~r~orl4dl~ a~ r~p~r~tlon
of 11po~o~o~, Vol.l Al~o ~o~ U S P~t~nt No. 4,a3~,~71, 1~-uo~
to P~p~h~d~opoulo~ ~t ~1., 1neorpor~t-d h~r~1n 4y r~rsnee to
th~ oxtont th~t lt t~4ch u th~ prop~r~tlon o~ 11POJO~H~ u~oful
~e~ordln~ to ~hc pr~n~ 1n~ntlon u d~-~r1 h d ~b w -
.
~ t hu ~l~o ~n ~ount. ccor~ to t~lo pruMtlnv~ntlon, th~ llpo~ hteh ~r~ r~l~ttv~ly ~ at 1~, iO
to ~0~ n-nom t~r~ 1n ~ t-r, pr~hr~bly 1--- th~n lO~ nm
~n dl~t~r ~hlblt m~x1~1 ~lount ~lt~ cwcr~je tn co~p~rl~on to
tho l~ger ~LV-type ll~ ot ~00 to ~0,000 n~ 1n d~t~r.
. .. ~ .
Th~ prof~rr~t llpo~o~ conpo~1t10n of tb~ pro~ont
l~ n ntlon co~prl u ~: pho~ph~tl~lcholl~o (lO colx)t eho
~ t1 nly ch~r~o~ pho~ph~tldyl~l~c-rol ~0 ~ol#), ~lth h th
pho~phollpld~ eont~ln1ng ~tyl eh~tn~ of p~l~ltle ~e1d ln tho
po~1tlo~, ~nd ol~1c ~C1t tn tho 2 po~ltlon~ ~d ehol~s~orol (20 -~
~ol~ For ~x1uu~ ~ound ew~rn~ tho pr~ h rr-t 11po~om~ ~r~
20 t~ ~00 n~ 1n dl~Jotor, pr~tcr~bly lc~ Sh~n ~00 nm. Th~
productto~ o~ l~poso~ o~ th~ 100 nu 1~ ~ceo~pl~lh~ by
p~lng ~LY llpo~o~s thro~h ~ Mlero~lul~t u rR ~lcrotluldl~,
Corp , N-w~o~, M~ nd gh~lr ~1 u ~llt typlo~ ry 1
th~n ~oYt ~0~
,
~h~n co~po~t of pbo~ph-t1ty1cholln~ ~th ~tur~t~d ~cyl
-~ th~1n~, th~ llp~o~u D~ th~ pr~-nt tnv~ntlon h-v- ~n qu~ou~ -
l~qu1d-cry~t~111no ph~ u tr~n~ltlon o~ h ~w~on about 3~ ~d ~2
, ~
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S~NT ~;L~W~ ~RI~ OLA~ .,.i 4- 2-~ ;4~AM i 7~3~41124~ 3~1 13~8i3 8
; 3 ! ~ J 7
d~Qr~ eonttgr~do, pr~r~bly cl~o to th~ hum~n ph~lolo91c-1
t~p-r~tur~ of ~7 d~gr~- c~ntlgr~d~ ~h~ llpo~o~ - ot th~
pr~nt lnv-ntlon ~hl~h ~r~ eompo~d ot pho-ph~tltylcholtn- ~1th
~on~n~ cont~lnln~ ~cyl ch~n~, ~xhlb1t phn ~ tr~n~1t10n~
bolow ~ro tog h~ ccnetgr~d-, ~nd ~1JO ~ho~ oo~d blo~d~orp~
proportlo~ to ~ound ~lto~
nv-ntlon 1~ not 11~1tcd eo only pho~phollpld or
phosphol1pld/cholo~t~rol cont~1n1n~ llpo~, ht ~l~o compr1~s
pho~pholtp1t/cholo~torol 11~0~0~4~ cont~lnln~ polyr~r~ thU~
llpo~o~o-po1y~r co~bln~t10n- prov1d~ for for~ul~t1~n~ ~1th
~ner~od vl~eo~tt,Y ~n~ pol~ r ~hlch ~tll pro~ld~ for
lncruud V~C~S~ round tho l~po~o~ tl~l~ w1th
th~ tt~uos o~ tho ~y~ e~n h u~ pol~r(~ ~r~ ~dd~d to
for ~ co~po-ltlon ~tth ~ co~1ty o~ ~t~n ~out 5
cp~ lOOO cp~., prcf~r~bl~ bct~n ~hut ~S cp~. and 200 eP-.
of pol~r~ h1ch c~n ~ u~-d ~ccord1n~ to ~h~ m~thod~
of tht~ vont10n ~aelu~- n~tur~l pol~n wch u
:~ po~ cch~rld~ ,9. ~gln~t~, ~t~nh, odtf1cd C~llUlo~c~
u hy~roxyprop~l~thylc~llulou ~nd h~Ltro~w~thl~lc~llulo~
g~l~t1n~, ~l hmln, cu-tn. ch~to~n ~n~ eoll~n. In add1t1u~,
synthot1c pol~r~ ~hlch c~n ~ d lnclu~ polyl~ctld~glycol1d~
eopol~r, pol~l~ctlh, polyh~rox~ th~l~th~cryl~t-,
pol~torullltu, pot~orthc~t~r~, pol~th-cr~ t-,
pol~lnyl~lcol~ol, polyvln~lp~rrol1don-, poly~er~t 16 ac1t,
poly~tbyl-noglycol ~lt pol~rlc u~ C1d~. Tho 11po~ ln
tho~r vl~eou~ ~trlx prov1do tor lon~r ~ontlon of th~
for~ tlon ln th~ ~ ~nt th~ or~ ~10~d~0rpt10n ot th~
llpo w ~ ~ to th~ 1t~l oY~r 10np r t1~ p~r10d~
p~rccnt~g~ o~ po~yn~r 1n th~ 11po wn~ ~or~ul~t10n W111 ~p~nd on
thC poly~or 1t~olf, ~t 1n ~nrr~l th~ conc~ntr~tlon o~ polyn-r
r'.'l c~n r~ngc tro~ ~out o.a~ ~ t~o ~t.~. -
) ~ :
::
~NT Ol';L~W~ PR~ L~BLAI~ ... i 4- 2-Y~ AM i 7~841124-' 3~1 13~
20137't~
Thor~p~utlc ~en~ ~hlch c-n ~ ro~ ~o ~und ~1t~J
ln llpoJo~ or 11po-o~-~po1~-r ~or~Yl~t1on~ ~h1eh blo~d~orb to
uch ~1t-- co~prls~ ~11 thcr~p-ut1e ~nt~ uu r~ ~or th~
tr~tm~nt o~ ~ound~, eut~ ~nd ~r~lon~ c-u--~ br ln~ur~, tr~um~,
~ur~ry nt/or dl~ ueh thor-p~ut10 ~gcnt~ ~111 e
comprlu, ~ut ~ro not lt~1to~ to~ oro~th ~ctor~, ~torot~,
~ntlox1d~nt~, ~on-t~rold~l ~nt11n~1~r~tory ~g~t~, ~ntlblotle~,
l~no~ul~tor~, ~nt1~ rglc~ ~nd co~poun~ h IlIk~ uP
h~nt ~br~n~ uch ~ ~lbron~ct1n ~ . Furth~r -
: g~t~ lneludo ~lologle~l r~pon~ ~odl~l-n ~ gon~ ~h~-
ch~n1-~ o~ ~ct10n 1n wlv~ tho 1ndlv1du~ o~n ~to~o~le~l
r~po~3~ , Ol~hu~ L~ , Ch~pt~r ~,
- ~or~nc~ c~du~1e ~s-, l~5, ll~c4rpor~t~d h-r~1n ~r
,,,
k eord1n9 tO th~ pr~n~ lnv-ntlon, th~ 11PO~O~J
cont~lnln~ th~ th~r~pwt1e ~gcnt or ~rug tor th~ tn ~te~nt o~
wound~ ~r~ ~ppll~t top1c~ to tho ~po. As ~lll bo ~nt~r~too~
by tho~o ~klllo~ ln th~ ~t, th~ ~du1n~tr~t~on, ~qu~Rc~ o-
~d~tn1~tr~tlon ~hcn ~or~ th~n ono thor~p~ut1e ~nt 1~ u~-d, ~nd
th~ eo~eontr~t10n~ ot th~ th~r~pwt10 ~o-nt- 1n ~hc ltpo-om ~ o-
th pr~ont lnvont~on d~pon~ on nYo~roUt ~ctor~. ~h~ f~ctor~
c~n 1ncl~d ~ th ~poG~ thor p-utle ~I~ or ~o~t~ to h u U~
tho nIt~r~ 0~ th~ ~ound. ~nt v~rlou~ el1n1c~1 f~ctor~, 1noludtn~
'~ th~ ~xt~ nd qp~ of ~ound h1~g tr~N~-d, th~ ~H41e~1 hltt~ry
î~ of th~ p~t1~nt ~ ~pto~ oel~t~ ~lth tho ~ound, ~llch
1n~1~mn~tlon or ~d-o-, ~t~. t~l~ctlon of th~ ~p~lflc
th~r~poutle ~p nt ln th~ 11po~ . th~lr conc~ntr~tlon ~nd
-~ ~oqu~nc- o~ d~ll w ry to tho ~y~ ~111 h ~d~ ~y tho ~k~llod
G1~n ~11d~ th~ for~go1n~ d~Jer1et10n. Jn ~ddttlon,
b~ uu~ th- llp~-~s~-typ~ ~or~ul~tton ~1c~d~orb~ ~nq/or
b10~dh~r-- p~e~1c~ t~ th~ wound ~tt~, th~ ~ount ~nd
~- ~r~qu~ney o~ th~r~p wt1c ~g~nt nor~ tl~ ~dJlnl~ter~t u th~ tr~
~gent ~-n b~ r~due ~. Typ1e~11y, th~ ~ount o~ ~hor~p~utlc ~g~nt
,' ,:
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S~NT ~Y:L~\'It PRlC~ L~LANC .,. ; 4- 2~ :5~AII; ~030~41124' 3~1 13~8;J~10
~' ~J 2~1377~
-
~d~1n1~t~r;d ln 11PUO~ ec~rd1ng to ~ho pr~-nt lnvontlon c~
b~ r~ue~ by up ~o ~ut ~ th~ ~o~ ~ln1~t~ oth~r
~tho~ uch u 1n ~quoou~ drop~, o1nt~ntl Ot ou~pon~10~
~ h~ l'ollowlne ox~pl~ ~r ~ ct~d ~o th~ blnd1n~ of
11POJOII~ ~n~ llpo~ prl~ hor~p~utlc ~g~nt~ to corn~l
~oun~. r~ x~pl~ u-tr-t1n of th- pro~orontl~l
b~nd~ng o~ th~ llp~ to Itou~d olt~J~ ~ut ~hoy ~ro ln no ~y
1 1~1t1n~.
.
~ Phosph~tldyleholln~ ~ ~.U q/~l~
Pho~ph~t1~ ol
Cholut~rol 0.~
Dloet~d~t ~ ~t~tr ~ thyl.lndoc~rbocy~n~n~ ~ ~ o) 0.1~ ~ /~1 .-
:
t
' '
A11quot~ o~ pho-phol1pld- ~nd chol~torol 1~ chloro-or~
22.~ ~ pho~ t1d~cl~ol1n-, N.5 ~ pho-ph~t1~1glyc~rol, ~.
m~ chol~torol) ~ro ~ddd to ~ ~lu~ eont~1n~r. th~ ~luor~c~nt
. dy~ ~0.11~) ~ th~n ~d~ ~ ~n ~ uo~ ~n ~thonol. th~
:: ~o~-nt ~ th-n o~p~r~ct undcr rlltro~ h~ llp1
~uoruecnt dY- t`1h n~ th~n roh~r~t~ Y1th Dulb~eeo'~
pho~ph~to buff-r~ ~lln~ cont~lnln~ ~ e~ nd 0.4~ IIM
~gnulu~ ~lO ~ h~ p~ ~u ~wt~ to ?.~ ~1th d1~ut~
Xl~. ~h1~ r~tlon r~ul~d ln th- ~pont~n~ou~
for~tlon of ~ult~lu~ll~r llpo~
}'
~;~ Fro~hl~ tl~ t~ r~lt cornoi~ hc~ 1n ~SS
~lu~R ~leon L~hr~orl-J. Ine.). rh- corn~ ~ro pl~c~ ln
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StNT Br;~W~ ~RlC~ L~BI.AH~ . . . i 4- 2-~ ;51AM i 7~3~41124~ 3~1 1398;~11
~J rJ ~ 1 3 7 7 ~
pl~tlc cu n ur~ dl~ho~ ~nd o1thor cut or ~er~pcd ~odl~lly through
thc cplth~ ur~C~ wlth ~ ~c-1p~ do. 1~11t~
~ollow1n~ th~ cuttln~ or ~cr~Plng, l~ul o- ~h~ ~luorc~ccnt
l~ol~t llpo~ torlnul~t1~n w~ ~ppl~od to tbo eorn~l Jur~e-
Afhr ~ ln tho Corn-~- wcro wUh~d ~ tl~ ~1th ~ SS ~lwR
oC ln ~ C02 lncub~tor u~ln~ ~ rot~r~ ~h~r.
E~ch corno~ w~ ox~ln-d u~ln~ tluoro~c~nc~ mlcro~copy
by vl~wlng tho cornu- in ~ ch~r cr~to~ by two 91~
eovor-l1p~ tlttC~ ln ~ ~chln~d plUtlC f~ow ull. ~h~ flod c~ll
con~1-t~ of ~ pl~Stlc ~lock ~11 5 X ll.t e~ X l.t cm) ln whlch
tour 1 c~ hol-~ wor~ bor-d for vls~ln~ und~r ph~c. A r~c~ d
~rc~ -2.5 X 2,5 cm ~nd -a.5 mm doop ~ - u ch1nod ov~r ~nd ~roun~
o~ch hol- ~n~ X IS ~m cov~r~llp ~dh~r-t ~1th s111con
to th~ block wlthln th~ roc u s~d ~ro~. A th1n ~h611~w ln~-t
tg~ -0.~ p w~ chlno~ ~1thln th ~.5 X t.~ c~ r~c~J-d
~r~ for ~ceo~o~t1nS ~ top eo n r~l1p of 2~ X 25 ~ tdhot~d to -
~hc 1-~9- ~lng ~lllc~n gr~o. On t~o oppo~lto d~ u o~ thc
block, ~our ch-nn~ p~r ~ , n rc ~chln~d ~lowtn~ ~or
pl~c~ nt o~ ~yr~ng~ ~odl~ ch tlp ~u~t ~nt~rln~ onc o~ ~ch
o~ tho 2 ~ cr X ~.~ c~ X 2.~ ~ r~tol~-d eh~rb~r~ E~gh ~yrin~
n ndlo ~u h~ld ln pl~co ~lth ~ scr~ n -h-r t~x~d 1n th- N ock,
P~r~ t~ ~n~r~ thc ch~b~r~ th nu~h thc ~yr1n~ no~dl~ In
th~ cont~r of tho ~lock tnD lon~ ch~nnol~ c~ X I cr ~n~ o~-
cm d~p) 1~ n uld conn ctlon ~lth tho ch- b-r- ~nr~ ~chln~d
p~rpondlcul~r to thc ch~nnol- ho~1n~ th~ ~r~n~ noodl~ for
colloeS10n o~ por-u--t-. Durln~ cx uln~t10n tho nO~ c~
flxod to th~ ~t-g~ of th~ ~lcro~copc ~lth ~cr-w~ -
. ~
~ ch ~ount-d cornc~ ~u ln ~urn pl~c~d on ~n l~ X ~8 m~
cwcr~llp 1n on~ ch~bcr of th~ n~ 6011 ~n~ eo~orod wlth ~ 2~ X
25 mm top con ~11p Durlno oxu~ln~tlon thc corn~ u ~Hr~
con~t~nt1y por~uud ~1th ~S~ ~lu~ ~nd tho to~p~r~ur~
e~lnt~1n~d ~t ~C ~1d~o 1~o~ o~ tho ~luor~eont~ el~d
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s~Nr ~:L~WE PRl~ Lt~LANC .,. . 4- 2-9~ ;lU:SlAM; '1~3~41124~ 3~1 139~;~12
~ V ~01~770
llpo~ pronn~ on thc corn~ ur~c~ ~ro r-~ort~t ~t -
~ynl~lc~tlon o~ 113~X. ~hc l~o~ ~ppo~ro~ bl-cl~ ~nd ~h1t- ~lth
th~ to r~lon~ r~prountln~ tho~o ~ro~ ln whl~h tho 1-~1cd
11po~o~ wer~ loc~tod. ~ho ~l~ck ~ro~ ~n tho 1~J
r~pr~cnto~ ~ru~ ~h~r~ no 1~hl~t llpo~ or pru-nt. On
Db~or~lng th~c 1~ o~, 1t ~ o~nd th~t th~ d llpo~
w~r- l~c~t~d w~tt~ntl~ll,v ln only th~ c4t er ~cr~p~ ~ltu.
Llttl~ to vlrtu~lly n~ l-hlcd lipoto~ ~h-Pod to th~ uneut or
un~cr~p~ ~r~ o~ ~ny o~ tl~- c~nn~. K~r w~ hl~
llposo~ ob~rv~d ~h~r1ng eO th~ corn~ n un~und~d or
"nor~-l" c~ w~ trc~e~ ~lth l-b-l~d ltpo~ ul~l wuh~
~co~dlng to tho ~bovo d~erlb~d ~roc~d~r~.
Tho l~p~ ~r~ th~n qua~t1t~t~ ~or t~ ar~ of ~hlt~
r~glon (1.~ hl1~ht~d ~tx~ hlch r~pr~Rt~d th~
blo~dsor~t llpo~omo~. ~h~s~ r~6ultJ ~r~l ~Xpl' J~d ~ corn~
sur~ rc~ (~a) t~ 3ho~n ln tho Fl~luro ~1th ~n ~nrag~
~t~nd~ rro~ ~ th~ ~n for ~lv~ dl-~sr flt 1~
3n tho ~1~ur~ 1t c~n b~ n ~hat ~u~ ntl~1tJ~ nD
l~b~l~d llpo~s ~h~r~d tO th~ UI~CUt or l~n~cr~p~ norul
oorn~ h~ l-b-l~ 11po~o~s lldh~ to about ~o ~ n
th~ ~ut eorn~ And ~ ~ ln tho Jer ~d corn~. Th~r~hr-.
llpo~ u-~ tcor~tn~ eo tb~ ~thod~ o~ th~ pr~nt lnv~ntlon
c~n bo u~ to dnt~v~r ~oun~ tr ~t1n~ ~pnt~ to ~ound~ b~us~
th~y prof~r~ntt~lly ~th~r~ to ~lound ~ltu ~nd not to unwound~
~tt~.
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S~NT ~Y;L~JW~ PRIO~ Lt~LAN~ .. i 4- Z-~8 il~:S2AM i 7~388411~4-' 3~ 8;~13
V ~ 3 r~ l7 ~3
Th~ ~o~ no c~po~lt10~ c~n b~ ~ppl10d tOp1C~ to
tho y- to promot- r ~plth~ t10n o~ ~ ~ounet-d cor~lta.
Pho~ph~tl~ylchol ~n~
,~ Pho~ph~tldy~ol~ rol 1
- Cholo~t~rol 8 mg
Ep1t~ 1 6ro~h f~etor ~E~F) lo ~
... '
For th- pr-~r~t10n o~ 10 ~1 o1' co~ ltlon, th~
pho~photlpld~ and cholest~ r~ ~t~h~ a~ ~ry lybphlll~
po~d~r- 1n 1l 9l~ cont~ln~r ~a30 q pho~h~t1tylcho11 0 m~
pho~ph~ttdyl~ rol, 50 ~ chot~t-rol) ~ollo~ ~y th~ adt1tlon
o~ l ~1 t hlt~nol. ~h~ r- t~ th~n ~olubllt2~t by g!~tl~
.~ Inlxlnp ln ~ ~0C ~ ~r b~th, ~t-rll~ ~tlt~r~ n~ th-n ~r~u
- tr~t to r~nw~ ~ha t ~ut-nol. ~h~ oph111~ 1d po~d~r t~
-~ th~l~ r~hydrat~d ~t1th 1~0 ~ of ~or11- E~P ~ olv~ 1n
C~O~ pho~h~ u~ d J~lln- ~ont~lnlng 0.9 rM c~lc1~n
.n~ ~nd t.~ ~ n~n-~1u~ ~10 ~ h1~ r hYdr~t1on ~ lt ln
th~ spont~n~ou~ for~tl~n of ~ultll~tl~r 11po~ co~t-1n1n~
. E~F. ~o ~x~ z~ ~n~r~ Hnt o~ EQF 11~ tho ltpo~ h6
~ormul~tl~n 1s ~rn ~- dr1~ c~n~ tl~o th~n ~h~dr~t~ wlth
. ~lbl ~t~rll~ do10n1~ t~r ~or~ UJC.
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:i~N~ ~r.~we I'R~e Le~LA~ .. i 4- 2~ ZAI~ i 7~3~0~ 4' 3~1 13~ 14
(--' U 2013770
-
f~tlo~lng t~ ~n ~xullpl~ o~ ~ llpo~o~/pol~r
~nul~t10n to llhl~h th~r~p~ut1c g-nt~ c~n b~ ~dd~ tor ~h~
tr~t~nnt o~ corn~ ount-
In~
~p~1~1toyl~2 ol~o,Yl~
phsJph~t1~1chol1n~ onnul~tlon
," ~.P~l~ton-t-ol~y~ .
pho~ph~t1dyl61yc~ t .~ or~ t10n
ch~ torol 0.~8 ~l ~or~ul~tlon
g~l~tll~ t~ o~ul~t10n
: .
e~
Th~ ~ho~hollp1~- ~ cholo~t~ h~ ~ t~y
lyophlll~ dors 1n ~ eontnln~r fol~ h~ ~tld1tlon
~f ~ ry~anol l~out ~ ml/J ~ o~ to~l llp1~. Ltp~phll~c
tru~ ~nt th~r~p-ut1e ~nt~ ~hleh ~r~ ~olllbl~ 1n t~r~1~ry
but~nol e-n bo ~ t th1~ po1nt ~t eollc-ntr~10n- ~ct1n
~r th~ tn~t~nt o~ ~un~. T~ ~1xtur~ h~n co~plat~ly
Itolub111~ golltl~ ~lxlng ln ~ ~0C nt~r l~ath and th~n fr~uo
drlod to r~ovo th~ t~r~ but~nal . ~h~ po~d~r 1
r~hydr~t~d ~1th Dulbocco'~ pho~ph-t~ ~u~ d ~lln~ eont~lnln~
L ~ o.~ ~ o~ n~ 0.4~ gn~ . Tht pH 1~ ~J~u~t~d to ~
~- wlth d11ut~ ~odtu~ hYdnx1t~ t~r tol4bl~ drug~ ant
~-` th~rapwt~t ~nt~ ueh u gro~h ~etort. c~n ~ ~W~d ~e thls
~; po1nt Jlt eoncentr~tlon~ nft~etl~ r ~h~ tr~nt of ~ound~.
~ult~l~llu 11po~ cont~lntn~ ~ru~ or th~r~p~utle ~nt~
. ~r~ for~d by lncub~lt1O~ ~nd ~1~tlng ln ~ roe~ry nt~ b~th ~t
0C~ For ~ ntr~ t ~ tcr ~olubl~ d~gJ ~n~
h~r~pcut1e ~ tt, th~ ~olYt1on c~n b tn~ rl~ ~ln ~nd
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5~NT Ol;L~We F'R~ L~ISLAI~; .. i 4- Z-~ 3AM i 7~3t841124~ 301 13~8i~15
3 7 7 ~
--
o~1n n~ r~h~dr~t~d w1th ~t~r to~lo~d by ~urth-r
1ncub~tlon ~n~ ~lx1n~ ln ~ rot~ t~r b~th ~t ~0C. . I~ ~m~ r
unll~ r llpo~o~s ~ro ~lr~d tho oultll~soll~r 11po~o~ c~n
b~ P~Ud tllrouo~ ~ MlcrofluldlurR (~lcronuldlc~. Corp.
rop~t~ly unt11 tho d~lr~d pr~por~lon of
obt~1n~. ~ ppr~ tod by tho-~ klll~ th~
Jom~ ltpo~ol- for~ul~tlon~, for ox~mple, ~ouo of tho~ cont~lnln6
prot~ln~, ~houl~ not b~ p u ~d throug~ th~ ~lcro n u1~1 u r b~c!u--
of d~n~tur-tlon of tho prot~n. ~ t stop, po~d~ro~ g~l~tln
(S~n~ ln typ~ I, Slg~- Ch~41c~1 Co., St. Loul~, ~0.) 1~ t
~t ~ eoncentr~t10n o~ ~5 n9/~l formul~tlon. 'rho ~1x~ur- 1~ th-n
;~ntly ho~too ~n~ ~lx~d ~t ~Dt u~t11 tr~ pl~t1n 1~ ~olub1111~d.
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