CA2118917A1 - Composition for the preservation of organs - Google Patents

Composition for the preservation of organs

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Publication number
CA2118917A1
CA2118917A1 CA002118917A CA2118917A CA2118917A1 CA 2118917 A1 CA2118917 A1 CA 2118917A1 CA 002118917 A CA002118917 A CA 002118917A CA 2118917 A CA2118917 A CA 2118917A CA 2118917 A1 CA2118917 A1 CA 2118917A1
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CA
Canada
Prior art keywords
solution
composition
preservation
organ
transplantation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
CA002118917A
Other languages
French (fr)
Inventor
Steven M. Strasberg
Robert P.C. Harvey
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mount Sinai Hospital Corp
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
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Publication of CA2118917A1 publication Critical patent/CA2118917A1/en
Abandoned legal-status Critical Current

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Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02Preservation of living parts
    • A01N1/0205Chemical aspects
    • A01N1/021Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0226Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients

Abstract

A novel composition for the preservation of organs intended for transplantation and a method of preserving organs intended for transplantation using the composition are described. The composition contains glucuronic acid or a physiologically tolerated salt or ester thereof in a pharmaceutically acceptable organ preservation solution.

Description

W093/04578 PCT/CA92/0038g .~ .
- - 21~917 Title: CONPOSITION FOR THE PR~SERYATIO~ OF ORGA~S

~AC~GRO~ND OF THE INV~NTION
The present invention relates to a composition for the preservation of organs intended for transplantation and a method of preserving organs intsnded for transplantation using the composition.
Tran~pIantation of organs is now considered to be a definitive ~reatment for patients with end stage liver, kidney, heart and pancreas disease. There is thus a great deal of interest in improving ex ViYo s~orage of cadav~ric organs and thus the viability of organ transplants.
The two most commonly used methods for organ preservation are hypo~hermic ~torage and continuous pulsatile perfusion. With hypothermic stora~e, the organs are rapidly cooled immediately after removal from the cadaver donor using a combination of external cooling and a short period of perfusion. ~he hypothermic storage me~hod is a preferred method due to its practicality and the ease of transportation of the organs. Continuous pulsatile perfusion involves hypothermic pulsatile perfusion after flushing with a chilled electrolyte solution.
A number of organ preservation solutions have been developed with a view to extending organ preservation time. Ringer's lactate and isotonic saline solutions have been used as extracellular flushing solutions and have been reported to allow for safe renal pr~servativn for short psriods of time, i.e. up to four hours. Storage for longer periods of time may result in severe histologic ischemic damage and subsequent non-function of the organs (column S, U.S. Patent No. 4,920,004) An intracellular electrolyte solution developed by Collins et al., (Lancet 2:1219, l9h9), has been report~d to offer se~eral advantages for hypothermic SUBS~TU~ Si~EET -:

.. . .. . ..

W093/04578 ~ t 1~ 9 1' PCT/CA92~00389 - 2 - `
storage, Table I shows the composition of the Collins solution which is most commonly used. ~odified Collins' solutions have also been d~veloped for use in hypothe~mic ætorage. For ~xample, Euro-collins solution is sLmilar to Collins solution with the exception that it does not contain magnesium.
Other organ praservation solutions which have been developed inolude Sacks~ solutions (S1 and S2) which have high i~tracellular ion concentration and osmotic pressure (Sacks, S.A., ~ancet 1:1024, 1973). Table I
shows ~he cQmposition of the Sacks-2 flush solution. The -solutions have b~en report2d to proYide Lmpro~ed transplan~ation results after ~torage of kidneys for up to 72 hours.
1~ Protecti~e additives such as ATP-MgCl2, AMP-Mg-Cl2 and ino~ine have also been included in pre~ervation/flush solutions. ~Siegel, N.J. et al., Am.
J. Physiol. 245:F530, 1983: Stromski, ~.E. et al, Am. J.
Physiol. 250:F834, 1986; Sumpio, B.E. et al., Am. J.
Physiol. 24?:R1047; Stromski, M.E. et al., Am J. Physiol, 250:F834, 1986). Belzer et al., (Transpl. Proc. 16:16I, 1984) deYeloped a perfusate containing ATP-~gCl2 ~see Table I for ingredients) but it has not been used for simple cold storage (Arch. Surg. 122:790-794, 1987).
U.S. Patent No. 4,920,004 discloses a hyp~rosmotic intracellular flush and storage solution that is reported to combine the salient features of Belzer~s ATP-MgCl2 perfusate and the simplici~y of the most commonly used Collins' C-2 Flush solutions. Mannitol is 3ubstituted in place of dextrsse in Collins' C-2 solution and adenosine and magnesium are added to the solutian to improve ~he preserYation properties of the flush solution.
U.S. Pa~ent Nos. 4,798,824 and 4,873,230 di~close solutions for the preservation of organs (particularly kidneys) prior to implantation, containing a ~pecific synthetic hydroxyethyl starch in place of ~erum albumin tD produce the required oncotic pre~sure. In W093/04~78 PCT/CA92/00389 ~; _ 3 _ ~ L7 particular, U.S. Patent No. 4,798,824 disrloses a solution including 5~ hydroxyethyl starch ha~ing a molecular weight of from about 200,~00 to about 300,000 and wherein the hydroxyethyl starch is substantially free of ethylene glycol, ethylene chlorohydrin, ~odium chloride and acetone. U.S. Patent No. 4,873,230 discloses a solution containing hydroxyethyl .tarch having a molecular w~ight of from about 150,000 to about 350,000 daltons, a d~gr~e of substitution of from about O.4 to about O.7 and being subs~antially free of ethylene glycsl, ethylene - chlorohydrin, sodium chloride and acetone. ~ Mar~hall's i30tonic citrate solution (Table I~
ha~ been reported to be capable of prolonging the psriod of ~afQ hypotharmic organ storage. It contains ~h~
impermeant anion, citrate and other substances which sre belieYed to pre~ent free radical injury.
U.S. Patent No. 4,879,283, di~close~ a solution for the pre~ervation of organs ~hich contains lactobionate and raffinose and has a solution ssmolality of about 320 mOsm/L, R~ of 120 mM and Na~ of 20 mM. The solution al~o csntains a synthetic hydroxyethy~ starch and other components such a~ glutathione and adenosine. The solution disclosed in U.S. Patent No. 4,879,283 is commonly known as the University of Wisconsin solution or UW solution and its composition is shown in Table I. The solution has been reported to successfully pre~erve the liver (Jamieson, N.V. et al., Transplantation 46:517, 1988), kidney (Ploeg, R.J. et al., Transplantation 46:191, 1988), and pancreas (wahlberg~ J.A., Transpla~tation, 43:5, 1987). Preliminary animal studies suggest that the solution ma~ al30 be effecti~e for the preservation of the heart (~icomb, W.N., Tran~plantation 47:733, 198B; and S~an~on, ~.~., J. Heart ~ransplan~ 7:456, 1988).
The UW olution and ~ar~hsll's i~otonic citrate ~olution are believed to provide improved organ pre~erYatlon a3 a re~ult of th~ir ability to pre~ent cell welling or oxygen free radical-mediated in~ury W093/04~78 '~lR.~ PCT/CAg2~0~3~9 (Belzer, FØ and 50uth rd, J.X., In: Transplantation:
Approaches to Graft Rejection. New York,: List, 291, 1986; Toledo-Per~yra, L.~. et al, Ann. Surg. 181:289, 1975; Downes, G~ et al, Transplantation 16s46g 1973; and Green, G.S~ Pegg, D.E., In: Pegg, D.E., Jacobson, J.A., eds. Organ preservation. Edinburgh: Churchill Livingstone, 86, 197g). The ~olutions contain impermeant anions, citra~e or lactobiona~e, which are added to maintain the normal double-Donnan equilibrium and pre~ent cell swelling in spite of the inactivation of the Na/~
ATPase -;dependent pump by hypothermia (Bslzer, FØ ~nd Southard, J.H., In: Tran plantation: ~pproaches to Graft Re~ection. New York: Liss, 291, 1386; Mar~in D.R. et al, ~nn-Surg 175:111, 1972; Southard, J.H. and Belzer, FØ
Cryobiology 17:540, 1980; and Mees N. et al., J. Trau~a 22:118, 19~2).
The pre3ent inventors have studied the relationship bat~een transplant ~iab~lity and liver function. In psrticular, the pre~ent in~entors u~ing a rat li~er model ha~e found that AST (aspartate aminotransferase) and LDH (lactate dehydxogena3e) concentration in perfusate, discriminated between ~iable and nonviable liver~ acros~ as well as within preservation groups. AST was found to give the best separation between viable and nonviable liYers. Functions such as ALT
(alanine aminotransfsrase) concentration in perfusate were found to separate viable from nonYiable liver allograf~s only within pre~ervation groups. In studying markers of allograft viability, the present inventors observed that rat li~ers stored at 1C for 4 hours or at 37C for 1 hour in a simple pre~ervation ~olution (NaCl 0.9%, CaCl2, 2 mN) wsré all viablo on transp}antation but those stored at 4C
for 8 hours or at 37C for 2 hours were nonviable. Cold pre~erYed nonviable livers were also shown ~o have increased vascular rssistance, platelet trapping and an initially low, but then high ri~e in AST upon reperfu~ion, W093~04578 ~ 7 PcT/~A92/oo389 all suggesting in~ury to the microcirculation. (Iu, S. et al, Tran~plantation, 44:562, 1987).
The present inventors ha~e further identified the morphological changes that occur in livers stored for the above-mentioned critical tLmas using light and electron microscopy after perfusion fixation. (~cReown~
C.N.B. et al., Tran~plantation, 46:178, 1988). Th~
pr~ent inventors ob~erved that r~Yersible injury ~a~
manifest by partial di~rupti~n of the endothelium a~d swelling o~ sinusoidal lining cells, hepatocytes appeared ` e~entially normal, apart from some minor bleb fo~mation.
Irrever ible in~ury and loss of viability w~s characterized by a completely deficiant endothelium;
lining cells were round with dark nuclei, and were detached from the underlying hepatocytes.
The present inventors also compared the effects on the microcirculation of preservation in UW and ~ar~hall~s solution and other control ~olutions and concluded that in~ury to the microcirculation was due neither to free-radical-mediated injury nor to cell swelling (Hollo~ay, C.M.B. et al., 48, 179, 1989~.
~U~NARY OF THoe I~VENTT~N
The pre~ent inventors have foun~ that inclusion sf glucuronate in a preservation sslution for use in the preservation of an organ intended for transplantation provides improved organ pre3ervation properties over other known preservation ~olutions. In particular, the present inventors have found that solutions containing glucuronats provided superior protection of the micrscirculation wh~n c~mpared to golutions based on noncarbohydrate or other carbohydrate anions in histologic studies of liver allografts. Further, results of liver function tests demonstrated that liver allo~rafts preserv~d in 8 preservation solution including glucuronate have greater transplan~ation viability *han allograf~s preserved in UW
~olution, preBeryation solutions in~luding glucurona~e may accordingly extend the preservation time of an organ - ~ r.~ . 7 W093/04578 21 . , ~ 1 1 PCT/CAQ2/00389 ~ v permitting increased organ availabili~y and sharing and decreased organ wastage which would necessarily result in reduced ~ransplan~ation costs. The use of a preservation solution inc~uding glucuronate may also provide better preservation ~uality for short term storage (i.e.
<30 hours).
The pre~ent invention therefore provides a composition for the preservation of an organ intend~d for ~ransplantation comprising glucuronic acid or a physiologically ~olerated salt or ester thereof in a pharmaceutically acceptable organ preservation solution.
Preferably the pharmaceutically acceptable organ preservation solution has ~ubs~antially the ~omposition of uw solution and most pref~rably is a modified UW ~olution de~oid of lactobionate.
The inv~ntion also relates to a method for pr~serving an organ inte~ded for transplantation in a sub~ect compri~ing i~fu~ing ~aid org~n with a composition comprising glucuronic acid or a physiologically tolerated salt or ester thereof in a pharmaceutically acceptable organ preservation solution. Preferably the pha~maceutically acceptable organ preservation solution has substantially the composition of UW solution and most preferably is a modified UW solution devoid of lactobionate.
D~S~RIPTI4N OF~ DRA~INGS
The invention will now be described in relation to the drawings in which:
Figure 1 is a photomicrograph of a section of a normal rat liver (original magnification x 400, H and E
stain);
~ igure 2 is a photomicrograph of a section of a rat liver allograft stored for 8 hours at 1C in solution~
containing normal saline and 2 mM calcium chloride (or~ginal magnification x 400, H and E stain);
~ igure 3 i~ a photomicrograph of a section of a r~ liYer allograft ~tored for 8 hours at 1C in a solution W093/~578 211 ~ ~1 7 PCT/CA92/00389 containing lactobionate at a concentration of 100 mM
(Lactobionate 1 ~olu~ion, Table III) (original magnification x 400, H and E stain~;
Figur2 4 is a photomicrograph of a section of a rat liver allograft sto~ed for 8 hour at 1C in a solution containing glucuronate, at a concentration of 100 mM
(Glu~urona~e 1 solution, Table III) (original ma~ification x 400, H and E stain~
~ EigNre 5 is a photomicrograph of a section of a rat li~er allograft Qtorad or 16 hours at 1C in a solution containing lactobionate 100 mM (Lactobionate 1 solution, Table III) (original magnification x 400, H and E ~tain);
Figure 6 is a photomicrograph of 8 section of a rat liver allograft stor~d ~or 16 hours at 1C in a solution containing glucuronate at a concen~ration of 100 mM (original magnification x 400~ H and ~ stain);
Figuxe 7 i5 a photomicrograph of a ~Bction of a rat li~er allograft stored for 8 hours at 1C in a solution containing isothionate 100 mM (Isothio~ate 1 solution, Table III) (original magnification x 400, H and E stain);
Figure 8 i5 a graph showing AST concQntration (mean + SEM) in perfusate of isolatad perfused rat liver (IPRL) reperfused for 3 hours at 37C after pxeservation in UW solution for 30 hours at 1C or for 30 hours at l~C in a UW based ~olution in which lactobionate wa3 sub~tituted by glucuronate on an equLmolar basi~ (GU~); after no preservation (NaCl - 2 mM CaCl2 pre~erved 6 hour~);
pre~ervation in ~urocollins solution (EC) for 30 hours;
and, pr~ervation in normal salin~ snd 2 mM CsCl2 fox hours;
Figurs 9 is a graph showing ALT concentration (mean ~ SEM) in perfusa$s of isolated p2rfused rat liver (IPRL) reperfused for 3 hours at 37C ~a) after preserv~tion in U~ solu~ion for 30 hours at 1C, or for 30 hsurs at 1C in ~ UW based ~olution in which lactobionate waC substitutsd by glucuronate on an , ~liS~17 ~quimolar basis (GUW); after no preservation (NaCl - 2 m~
CaCl2 preserved 6 hour~); preservation in Eurocollin~
solu~ion (EC) for 30 hours; and, preservation in normal saline and 2 mN CaCl2 for 8 hours;
Figure 10 is a graph sho~ing bile production (mean * SEM) in perfusate of i~olated perfused rat liver ~IPRL) reperfused for 3 hours at 37C after preservation in UW solution for 30 hours at 1C or for 30 hours at l~C in ~a UW ba~ed solution in which lactobionat~ was substituted 10 by glucuronate on an equimolar basis (GUW); after no ~ :
preservation (~aCl - 2 m~ CaCl2 preserved 6 hours);
preser~ation in Eurocollins solution (EC) for 30 hours;
and, preservation in normal saline and 2 mM CaCl2 for - hours; and Figure 11 is a graph showing platelst concentration (mean + S~M) in perfusate of isolated perfused rat li~er (IPRL) reperfused for 3 ho~rs at 37C
after preservation in ~N solution for ~0 hour~ at 1C, or UW for 30 hours at 1C in a UW based solution in which lactobionate was substituted by glucuronate on an equLmolar basis (GUW); after ~o preservation (NaCl - 2 mM
CaCl2 preserved 6 hours); preservation in Eurocollins solution ~EC) for 30 hours; and, preservation in normal saline and 2 mM CaCl2 for ~ hours.
DETAI~ED~D~S~RIPTION 0~ T~E I~VENTIO~
As hereinbefore mentioned, the pre~ent inYention relates to a eomposition for the preservation of organs intended for transplantation eomprising glueuronic acid or a physiologically tolerated salt or e~ter thereof in a pharmaeeutieally aeeeptable organ preservation solution.
The eomposition of the invention has Lmproved organ pres~rYation properties over previously known preservation solutions.
The terms ~preservation" r "preserving" or ~nfusing~ u5ed herein inelude but are not limited to perfu~ion, flushing and storage of an organ intended for ~ransplantation.

211~917 g ' ~

q.~r.e p~ar:~aC~utlC~Blly a^ce~-able nrB~n pre~er~a~ian 8Q~ 'On U~;elO. ir. ~e ~Qmpasi~lOn o~ rhe ir.ventior~ y be ~ny _o~moaiy u~ed ~r~6enr~tic~r. solutlon. The ingreà~ent3 cf exe~pl~-y _o~mon;y used p~ese~vat~o.n 30~ tiO;I~ e~ 61~" ~0"'~ ln r~ble 1. ~h~ ~clutlon 5 d~c;osed ~n 13,~ atent ~o. ~,B73,~a~ i~ c~Dmor.ly ~nD~n ~9 ehe - U~ivers._~ 3~ ~ scon~in ~oiu~ios o- UV 6s~1utlor~ ~nd i~s de~aiied rcm~ sit~on 15 eho~ ln ~b~e I. Pr~rY-~Dl~ ~he pre~e_vstion s~Lut~or.
has suD~tar.rlally ~h~ compo~itil~r. ~f UV ~oiu~i~r. !~oe T~ble I~ ~!Ln~
I:~DSt pr~ferabl~ is & modi~ied UU solu~ dev~id ~f lacto~ionate arld 10 c~r~t~ining ~lu~u~on~e is~ ~n equ~lar dose, : . .:
The ~ucur~r.ic acl~ or ~ phy~lolo~,ical~y '-clerated ~l'c Dr e~ce~ chereDf used in ~he ~posi~ion o~ tbE~ inv~llt~cn ~y~ be ob~a_ned u~ the ~ethod~ ce6c_ib~ by ~lehl.re~ter ~J~.dvan. C3rb~hydrate ~:e~nistry, 8:231, 195") or ~y De obta~ed ~om~ rc~a1ly ~for exa~ple, 15 fom S~ Che~D. Co., 5t. Lou~3, ~0, U~S.~ lte c~pou~ld3 th~t may he u~ed irL ~he compo6ition o~ tn~ inven~ion lnolude gl~u;onlc ~cid~
~odiun~ ~lu~uro~ate, pots6sluu: glucurc~te ~nd other ~luble g,lUOUrOnd1:B 3altg or e~er6. ~ha concsntr~lon of ~he glucu-oni-: ac:ld cr ~ p~y~lologic~lly tol~ra~ ed sAle or e~ter thare~f ln ehe 20 co~pwi~ion of th2 lnYentio~ m~y be ~ro3 2~0ut 3 to 2G0 m~, pr~fer~bly sbout 100 to 140 m~, :Do~t prafdr-bly 100 ID~

profo~ed ~bod~en~ ~f the i~v~ n, the comp3cltlon includes but is not ll~i~ced t~ the co~p~n~n~ set out T~bl~ II here~.

In a p~rtl~ular~y preferred e~bodimerl~ ~f the iD~renticn th~ co~o61tion co~pri~e~ 130 ~ glucurona~e ta~ ~dlu~ or potA~6iu~
3alt) 2S m~ X~4. 5 $~ ~gSO~î 30 ~ f no3~, S mM adYn~Y~. 3 m~
glut~ione~ 1 m~ ~lLopu~lno1, 5C g/l hydroxyethyl ~t~rch, lOQ ~
1n~ull~, 0.5 ml/l b-ct~1~, a~d 3 3gll dex4~0tha30~e. ~he coDposition o~ the inYe~tion ~y be prepared by ~i~in~ ~h~ v~rl~us co~pon~n~s of the co~po0ition uCing con~2n~onal me~hv~s. In partlcular, tha v~Soun co~po~n;s v~ ~h~ ~v~p~31~10n of Ch~ 1m ~tlon ~h~ bo ~lxed a ll~er of di6~11ed v~t~r to prod~c~ ~h~ oYer~ll conposition. Th~

SUe~;TlTUTE ~;HEET

~093/~4578 2 l 1 ~ '~ 1 7 P~T/CA9~/003~9 preferred composition of the invention may be prepared ac~ording to the constituent ranges set forth herein in Table II or accoxding to the preferred amounts ~et orth above.
The solution may be brought to a pH of about 6~B
to 7.6, preferably 7.2, at 25C with NaOH. The ratio of Nat to R~ in the resulting composition may ~e about 150:3.5 to 20:140, preferably 30:120 which is the ratio of th~s~
ions in UW solution~ Osmolality of the resulting snlutions may be ad~usted to about 300+30 mOsm/l using, if desired, variable concentrations of mannitol.
The in~ntion also rslates to enhanced embodiments of the composition of the invention whi~h - includ~ the composition described above containing oth~r additive~. Examples of su~h additives are enzyme inhibitors (e.g. 2'-deoxycoformycin), buffers (R~C03) and calcium entry block~r~
The present invention also contemplate~ a method for preserving an organ intended for transplantation through the use of the composition descri~ed aboYe. In particular, an organ such as a liver, may be flushed during harvesting and after its removal from the donor with a composition of the invention. The organ is then s~ored in a composition of the in~ention under hypothermic 2 conditions. AlternatiYely, after initial flushiny, the organ may be connected to a plLnp wherein a cold perfusate of the composition of the invention is continuously circulated through the ~rgan. Prior to transplantat~n the organ may be flushed again with the composition of the invention.
The method of th~ inv ntion may be! used ~o preserve any organ intend~d for transplantation, preferably an in~raabdominal organ such as the liver, pancreas and kidney, most preferably the liver.
The fsllowing non-limiting examples are illustrative of the pressnt invention:

,, , . . , ~ .

W093/04~78 PCT/CA92/00389 ;~-1 1 8 .
~:cam le 1 ~-A variety of pre~ervation solutions bas~d on different anions, both carbohydrate and noncarbohydrats, were ~ested for their efficacy in protection of the microcirculation using morphologic criteria. In particular livers stored in preservation ~olutions based on earbohydrate and noncarbohydrate anions w~re compared with livQrs stored in ~aline and i~otonic citrate, and ~ith each othar. Comparisons of solutions containing different carbohydrata anions were also made to identify the an~on that pro~ided the best pre~rvation of the microcirculation. T~ evaluate the importance sf hydroxylation of ths anion, livers stored in solutions ba~ed on butyrate, ~-hydroxy~utyrate t and y-hydroxybutyrate were also compared. The latter twocompounds ha~e a similar carbon backbone to citrat~ and have hydro~yl groups at carbons 2 and 4 r~spectiv~ly;
butyrate i8 not hydro~ylated and was used as a control.
The detailed composition of each pres~rYation solution tested i8 shown in Table III~ Predominantly univalent anions were used at a concentration of 100 mM, the same concentration of lac~obionate in ~W ~olution (Jamieson, N.~. et al, Transplantation 46:517, 1988).
Reduced concentrations of divalent anions wer2 employed.
Most ~olutions contained 5 mM MgSO~ although some other~ise identical solutions with 40 mM NgSO~ were employed to evaluate the efficacy of high or low concQntrations of magnesium. Osmolality was kept constant in all solutions at 300~3 mOsm/l, using variable concentrations of 3~ mannitol. The pH of all solutions was brought to 7.4 at 25C with NaOH.
Where possible, the ratio of Na~ to ~ in the t0B~ preservation solution~ was 45:54 which is the ratio o~ these ions in i~otonic citrate solution. For anions not a~ailable a5 the R~ salt or free acid, only the sodium salt wa~ used.

W093/Q4578 9 ~ 7 PCT/CA92/00389 The following techniques were used in evaluatin~
the preserva~ion solutions ~ ivers were harvested from male ~istar rat~
(20Q - 300 g), using the method for donor hepatectomy described by Kamada, N. and Calne, R.Y., Transplantation 28:47, 1979, and Transplantation 93:64, 1983. All livers were flushed in situ wi~h 12 cc of the test preservation solution to be tested with 500 i.u. heparin, followed by 20 cc of the preservation solution alone, then stored in the preservation s~lutions to ba tested.
Livers pre~erYed in the solutions to be tested were stored $or 8 hours at 4C and were compared to the appearance of livers preserved in saline or i~otonic ci~rate for the same time period. If the appearance ~a~
mo~e comparable to the latter, the experiment was repeated using a 16 ho~r st3rage time.
~ ollowing preservation, perfusion-fixation wa~
preformed as described in Mc~eown, C.~.B. et al, 1988, Transplantation, ~6.178. SpecLmen~ were then embedded in paraffin, sectioned, and stained with hematoxylin and eosir.. All slides were blinded prior to examination, and assigned a qualitativa and descrip~ive evalua~ion. To assist comparison of livers stored in different solutions, sections were scored on a scale of 0 to 8. A score of 0 described a liver with the appearance of unpreserved controls; 4 represented the ~ppearance associated with the limits of ~iability, and 8 the appearance associated with lethal preservation in~ury, ~ased on saline-stored controls described in Mcgeo~n, C.~.B. et al, Transplantation, 46:178, 1988.
Solutions associated with livers having a score of 4 or less, that is, a histologic appearance compatible with v~ability af~er 8 hours storage, w~re consid~red superior solutions.

WO93J04578 2 ~ 1 8 .~ :~ 7 PCT/CA92/00389 A. ~omparison_of So~utions Containin ~ arboh~dr~e v. Non~rboh~drate ~nion~
Livers ~tored in solutions based on propionate or butyrate and its hydroxyl derivatives or i~othiona~e, i.e. non carbohydrates (Table III) were compared with livers s~ored in solutions based on galacturonate, gluconatet glucoheptonate, saccharate, glucuronate, and lactobionate (Table III) i.e. carbohydrates. Sev~ral qlucuronate, lactobionate and gluconate so~utions were tested. In the~e ~olutions the amounts of sodium and pota~sium salts of the~e ~ubstances as well as the amount of ~gS04 was varied according to the specific formulae outlined in Table III. Li~ers pre~erved in solutions containing either a 5 mN or 40 mM concentration of ~gS0 were histologically indistinguishable.
The photo~icrographs of sections of rat liver allografts stored in thQ ~oncarbohydrate solutions ~se~
Figure 7) and in th carbohydrate ~olutions (Figure~ 4 and 5) were compared to photomicrographs of a section of normal rat liver (Figure 1) and sections of rat liver allografts stored for 8 hours at 1C in solut~ons containing normal saline and 2 mM calcium chloride (Figure 2). Figure 2 shows the well developed in~ury with rounding and detachment of endothelial cells.
Following 8 hours of storage, livers stored in solutions based on non-carbohydrates all had mean increa~es in weight of several per cent. The lobular architecture of the~e liver~ was well maintained.
Hepatocytes had variable degrees of peripheral vacuole formation and bleb~ were readily ~isible in the sinusoidal lumina. SLC had round, darkly-staining, py~noti~ n~clei, and were also usually detached from the underlying substratum pro~iding only moderate ^~mprovement in the picture ~hown in Figure 2. Figure 7 provides a repre~entative example of the effect of these non-car~ohydrate ~ol~ion3.

W093/04S78 ~ 1 7 PCT/CA~2/00389 LiYers stored in carbohydrate bs~d solutions had mean weigh~ lo~ses of several per cent. H~patocytes showe~ only rare vacuole and bleb formation, and most SLC
were attached, although swollen and even round and darkly-staining. Some detached cells could be se~n (Figure 3 and4). In particular, Figure 3 showed marked improvement in preservation when eompaxed to Figure 2, with only minor de~achment and rounding of endothelial c~lls. Figure 4 showed even bet~er preservation than Figur~ 3 with only very minor changes.
These compari~ons revealed that regardless of Na:R ratio, magnesium concen~ration, or anion concentration, livers stored in s~lutions ba3ed on carbohydrate anions were as30ciated with weight loss and a moderate degree of injury to SLC after 8 hours of cold s~orage, while ~hose stored in noncarbohydrate anion ontaining solutions gained weight and demonstrated more ~evere microva~cular in~ury histologically. Among livers stored in solution~ based on carbohydrate anions, the degree of liver weight gain did not relate to the morphologic appearance of the microcirculation.
B. ~ompari~on_of Various ~ar~h~drate_An.ions in Preservation SolutiDns Five different co~parisons ~are uced to identify the carbohydrate anion whose inclusion in preservation solutions pro~ided the best preser~ation of the microcirculation as determined by the screening test.
Solutions based on galacturonate, gluconate, lactobionate, glucoheptonate, glucuronate, and saccarate w~re evaluatsd.
Li~ers stored for 8 hours in solution based on galacturonate, lactobionate (lactobiDnate 2) and saccarate ~see Table III for compositions of ~olutions) ~ere compared. All contained ~gSO~ 5 mM ~ similar ratios of Na:K, impermeant anion concentration of 100 mN (except saccaratB) snd had final 08molalitie5 of approximately 300 mOsm/l. Saccarate wa3 employed at a concentration of 85 mM becaus~ it3 valenc~ of Z mandated high cation W093/~578 2 11 ~ g 1 7 PCT/CA92/00389 concentrations. Livers stored in ~he galacturonate, lac~obionate 2 and saccarate solutions had changes in liver weight of -1.2, 9.6% and -3.36~ respectively.
Galacturonate pro~id d significa~t protection of the microcirculation. After 8 ho~rs of s~orage in the galac~uxonate solution, lobular architecture was w~ll pr~served and hepatocy~es looked quite normal with few v~cuoles or blebs. SLC were s~ollen, but the ma~ority r~mained attached. Tho~e tha~ were detached appe~red swollen and rounded rather than truly pyknotic. In some areas, normal, flat SLC could be ~Qen. ~ivers stored in the lactobion~te 2 solution had nonmal appearing hepatocyte~. SLC at all ~tages of in~ury wer~ pre~ent.
Some appeared normal, -flat and elongated, some wsre swollen and others were detached ~ith round, darkJ
pyknotic nuclei. This appearance is ~ypical of livers stored to the limit of ~iabili~y, ~ determined in a previous study of saline-preserved li~ers (~cR20wn, C.~.B.
et al, Transplantation 46:178, 1988~. SLC of livers stored in the saccara~e solution, although not entirQly comparable because of the reduced concentration of saccarate used, wers less well preserved than thsse of li~ers stored in solutions based on galacturonate or lactobionate. N~st were round with darkly staining nuclei, and frequently were attached to the Qubstratum o~er a ~mall area of the cell surface. Occa ional swollen attached SLC were seen. Hepatocytes were normal ~nd ~acuoles and ~lebs were rare~
Liver~ stored in solutions based on gluconate, glucuronate and lactobionate (solutions gluconate 2, glucuronate 1 and lactobisnate 1 in Table III) wer~
compared. All 301ution~ contained MySO~ 40 mN, a final osmolality of 300 mOsm/l, an impermsant anion concentration of 100 mM and a Na:~ ratio of 46:54. Mean change~ in liYer weight after B hour~ of co}d storage were -1~5, -8.0 and -5.6% re3pectively. Li~rs stored in the glucona~e 2 ~olution had well preserved hepatocy~es with W093/04578 ~ 17 PCT/CA92/00389 blebs and vacuoles visible only in some sections. A
variable degree of microvascular in~ury was noted ranging from a typical pictu~e of reversible in~ury with ~wollen but at~ached SLC and few detached cells, to pxedominately detached cells wi~h round, darkly-staining, pyknotic nuclei. Livers stored in tha glucuronata 1 ~olution were notably superior (Figure 4). Hepa~ocytes were well-preserved, and freque~ntly had a granular cytoplasm, an appearance noted in an earlier s~udy af~er preservation in UW solution (Holloway, C.M.B., Transplantation 4B:179-182, ~ 1989). SLC were generally swollen, but all were attached, and flat; elongated cells were fsequently ~en. Livers preserved in the lactobionat~ 1 ~olution were associated with normal appearing hepatocytes and few blebs or vacuoles. Sinusoidal cells wera almost all attached to the underlying substratum. ~any were simply swollen, b~t a significant number were rou~d, with darkly-staining pyknotic nuclei (Figure 3). This comparison clearly Lmplicated an advantage to glucuronate over gluconate or lactobionate in the protection against micxo~ascular injury.
E~a3ple 2 The efficacy of solutions based on lactobionate was compared with those based on glucuronate. Co~parisons were made of solutions containing the principal electrolytes found in UW solution and solutions which are isosmolar glucuronate. Comparisons were also made of the lactobionate 1 solution and glucuronate 1 solution, and of the lactobionate 3 and glucuronate 5 solution. See Table III for composition of test solutions.
Histologically, li~ers stored for 8 hours in these Bolutions had a similar appearance. Lobular architecture wa~ well preserved, and hepatocyte blebs and Yacuoles ~ere rare. Almost all SLC were attached, but swollen, or rarely round and pyknotic. Occasional elongated, ~lat cells were observed. Similarly, after 16 hours of cold stsrage, little difference could be WQ93~04~78 2 11 8 917 PCT/CA92/00389 discerned~ although review of blinded slides by two independent observers consistently noted slight but definitely superior preservation with glucuronate.
Figure 5 shows substantial in~ury to the endothelial lining with rounding and detachment of endothelial cells.
Figure 6 shows less injury to the endothelium than in Figure 5. Architecture wa~ well preserved and bleb and vacuoles w~re prominent. Severa microvascular iniury was pressnt with most 5LC rounded with darkly-staining pyknotic nuclei~ Nany were detached, and others wer~
attached to the ~ubstratum over a small area of th~ cel~
surface. The SLC ln~ury seen in the~e liv~rs was 1~88 than that seen aftex storage for 8 hours in NaCl 0.9% and - CaCl2 2mN (Figure 2).
~ample 3 The uperior protection of the microcirculation provided by ~olutions containing 100 mN glucuronate over solutions ba~ed on noncarbohydrate, or e~en other carbohydrate anions prompted the evaluation of livers 20 stored in ~imilar solutions but with different concentrations of glucuronate. Livers were stored for 16 hours in solutions containing 100 mM glucuronate (Glucuronate I, Table III), 140 mM glucuronate (Glucuronate 2, Table III), and 200 mM glucuronate (Glucuronate 3, Table III). The 200 mM Glucuronate solution was mannitol free and had an osmolality of 366 mOsm/l. The Na:K ratio in all solutions was 46:54.
All livers stored in glucuronate based solut~ons lost weight during storage. The change in ~eight of li~ers stored ~n the solutisn~ containing 100, 140, and 200 mM Glucuronate for 8 hours at 4C wa~ -7.1, -5.29 and -4.5% respecti~ely and after 16 hours was -11.2%, -12.2 and -6.6% rs~pectively.
The histology of livers stored in the 100 mM and 3~ 1~0 mM glucuronat~ solutions were quite similar wherea~
liver~ ~tored in the 200 mM ~olution showed deterioration in the morphology of th~ sinusoidal lining cells.

W093/04578 ~ I 1 8 .q 1 ~ PCT/CAg2/00389 ~xample 4 Liver function~ in livers stored in U~ solution and a solution containinq glucuronate ~GU~) were studied using the isolated perfused rat liver ~echnique as described in S. Iu, P.R.C. ~arvey ~t al, 1987 Transpl~ntation 44(4):562. Th~ glucurona~e solution (GUW) was prepared as follows 19.41 grams of,glucuronic acid (Sigma) was added to 400 ml of distilled and deionized water at room ~emperature (20 to 24C) and mixed ~ntil ~he solution ~as clear of undissolved material. 20 ml of 5N
ROH solution, 4.0 ml of SN ~aOH solution, 1.34 grams of adenosine, 0.135 grams of allopurinol, 3.4 gram~ of potassium phosphate monobasic, 0.60 grams anhydrous magnesium sulfate, 17.83 grams of raffinose, 0.92 grams of glutathione and 500 ml hydroxyethyl starch (pentastarch) were each added to the miarture and the mixture was stirred af~er the addit~on of each constituent until the constituent wa~ di~solved. The pH of the ~olution was then adjusted to pH 7.4 + 0.1 with 5N ~aOH. The volume was adjusted to 1 liter and the solution was stirred for 5 minu~es. The UW ~olution was pxepared by the same procedure but lsctobionic acid (35.83 g/l) rep}aced ~he glucuronic acid.
Four liver function tests, namely AST (aspartate aminotransferase), and ALT (alanine aminotransferase) concentrations in perfusate, perfusate pla~elet count and , bile flow wer~ u~ed in the study. Livers were preserved for 30 hous at 1C in U~ or GUW and then reperfu~ed on the isolated perfu~ed rat liYer system at 37C for 3 hours.
AST and ALT were measured using commercially available kits (Boehring~r M~nnheim). Th~ results of the li~er fun~tion te~ts are shown in Figure 8 (AST), Figure 9 (ALT), Figure 10 (Bile flow) and Figure 11 (platelet count) and summarized in Table IV.
Li~ers s~ored in GUW showed marked and statistica}ly improv~d function in ~ST and ALT relea~
during the reperfu~ion period. The pre~ent in~entors have W093/04578 2 1 1 8 g 1 ~ ~CT/CA~2/00389 1 9 ' , ~
previously sh~wn that AST release is closely related to transplant viability in this model (Iu S., Transplantation 44: 562, 1987). The reduced release of the transaminases AST and ALT signifies reduced liver cell injury in the GUW
group. Toledo-Pereyra et al. (Ann Surg, 181: 289, 1975) have also found that the results of auxiliary transplantation were best when no increases in ~ST
occurred duriny perfusion. There wa~ also a significant reduction in the platelet count in the UW group bu~ not in the GUW group. This signifies that there is less micro~ascular injury in the GUW group, a conclusion supported by the morphological studies described above.
Finally bile flow was better in ths ~UW group although in this paramenter statistical significa~ce was not guite achieved. The results suggest that allografts preserved in the GUW solution containing glucuronate ha~e greater transplantation viability than allografts preserved in UW
solution.
The present invention has been described in detail and with particular reference to the preferred embodiments; however, it will be understood by one ha~ing ordinary skill in the art that changes can be made thereto without departing from the spiri~ and scope thereof.

3 lr7 WO 93SQ4578 PCI'/CA92/00389 - 20 - ~.
T~BI.E I
Composition of ~nown Preser~ration Solutions .
' '. ~`
Collins-2~ Sacks-2~ Belzer~ ~:
gJl g/l Perf7s~te ~I2PO, 2 . 0 4 . 16 3 .
K2HPo43~2O
RCl 1.12 - --~HCO3 ~ 2 . 30 Nannitol - 37 . 5 Glucose 25 - 1. 5 MgSO4/7~2O 7 . 38 _ 8 l~Cl2 ~ ( 2 meq/ml ) Adenosine - - 1. 3 :
Sodium Glutathione - - 17 . 5 P.lbumin - - 5 . 3 ~ :
NaHCO3 0. 84 }.26 -Allopurinol - - 0 .1 13 ~
Verapamil - - - -K~
~actobionate Citrate .
Raf f inc)se Hydro~yethyl starch - - -Osmolality ~-~mOsm/~g) 320 430 300 pH 7.00 7.00 7.10 ~ -* Cs~mpo~i'cion~ taken from U.S. Patent 4,920,044, colwnn 5, line 48 to cc~lumn 6, line 13.
:' ' .

W093/0457B~ 9: 17 PCT/CA92/00389 ~ 21 -TAB~E T - ~ont~d ~:~
Composition of Rnown Preservation Solutions ^.
U.S. Marshall~ UW~ -P a t e n t Isotonic g/l No.4,920,0 Citrats~ -04 ~m~) g/l .' .
RH2PO~ 1.54-2.56 3.4 ~HP 4 H~ 12.i ~Cl 0.84 - -1.40 KHC~ O.63 - -3 1.05 Mannitol O - 2.~7 100 Glucose 20 - 37 MgS04/7~20 o.o - 4.62 40 1.23 MgCl 0.0 - 0.4 0.3 2 meq/ml Adenosine 0.75 - 1.34 1.2~
Sodium 0 92 Gluta~hione Albumin -- _ NaHCO3 0.63 -1.05 AllopurinolO - 0.125 9.136 Verapamil O - 1.0 K' - - - 35.83 Lactobionate Citrate 56 ::
Raffinose - - 17.83 Hydroxyethyl starch Osmolality 255 - 425300 mOsm/l 320 - 330 mOsm/k~ mOsm/kg pH 7.29 - 7.1 7.40 7.50 W093/04578 211~ L~ 22 - PCr/~A92/00389 * Composition taken from Hollo~ay et al., Transplantation 48:
179, at 180, 1989 ** Composition obtained from Folkert D. Belzer and James H.
SoutAard, University of Wisconsin.

W0 93/04578 1 ~ ~ 9 1 7 PCI /CA92/00389 l~l~E II

Concentration mN
Glucuronate ( as sodium or potassium ~alt)3 200 RH2P04 5~40 ~lgS04 ~ 0 ~ .
Raf f inose 5_5 0 Adenosine 1-50 Allopurinol o . 5-5 . Glutathione 1-10 Hydroxyethyl Starc:h 10-70 g/l Insulin 0-100 U/l 0-1 0 ml/l Bactrim 0-1~ mg/l Dexame~hasone W093/04~78 ` PCT/CA92/00389 .81~ 17 - ~4 -TABLE ~II

Composition of.Test Preservation SolutiQns* `~

__0__________________________________________________~__ Name of Solution Butyrate 1 Composition ~ `
Cation Anion Concen~ration Conc. in -~
mM g/l Sodium Butyrate 140 15.414 Magnesium Sulphate 5 1.232 Vncharged solutes Manni~ol 25 4.55 Osmolality: 3~0.
________________________________________~________________ Name of 5Olution ~lpha-hydrQ~_putY~a~e_ Composition : -Cation Anion Concentration Conc. in ~
mM g/l .-Sodium Alpha-hydroxy ::
butyrate 140 17.654 Nagnesium Sulphate 5 1.232 ;
Uncharged solutes Mannitol 23 4.19 Osmolality: 300.
________________________--------_--_------------ ---- ---------- -- ~

* The composition of these solutions is listed according to original ingredients. Anions and cstions were often added as their respective acids and bases and ti~rated to neutrality. Therefore, to obtain final composition eliminate Hl and OH- as listed in the Table.

W093/04578 21 1 8 ~ ~ CA92/00389 TABL~ ont~d Nam~ of Solution Ga~a-~Y~ro~y butvrate Compo ition Cation Anion Concentration Conc. in mN g/l Sodium Gamma-hydroxy butyrate 140 17.654 Nagnesium Sulphate 5 1.232 .
Uncharged solutes Mannitol 28 5.10 .Osmolality: 300.
_____~_______________________________________________..___ Name of Solution Gala~turonate Composition Cation Anion Con entration Conc. in ~ g/l Sodium Hydroxide 46 Potassium Hyo`roxide 54 3.03 Hydrogen Galacturonate100 21.22 (Galacturonic Acid) ~agnesium Sulphate 5 1.232 Uncharged solutes 115 20.95 O~molality: 300.
_________________________________________________________ WOg3~04578 PCT/CA92/00389 2 1 1 8 ~ 1 7 25 ~

~ABLE III - Cont'd ~
'.' ' Name of Solution Glucoheptonate~
Composition ;~ :
Cation ~nion Concentration Conc. in ~odium Glucoheptonate 140 38.5 Magnesium Sulphate 5 1.232 :- -Uncharsed solutes ~-:
Mannitol 50 9.11 :-,--:: ::
Osmolality: 300. ~
Name of Solution Gluconate l ~:`
Composition ~-Cation Anion Concentration Conc. in mM g/l Sodium Gluconate 37 8.069 -;~
Po~assium Gluconate 19 4.450 Magnesium Sulphate 40 9.B59 ::
Potassium Chloride 25 1.865 ~:
Uncharged solu~es Mannitol 100 18.22 -~
Osmolality: 302. ::

W0~3/04578 ~ PCT/CA92/00339 T~ on~'d -: -~ame of Solution Gluconate Compositio~
Cation Anion Concentration Conc. in mM g/l Sodium Gluconate 46 10.032 Potassium Gluconate S4 12.646 Magnesium Sulphate 40 9.859 Unch;rged solutes 70 12.75 Osmolality: 300.
____________________~________________________0___________ Name of Solution GluconatQ 3 Composition Cation Anîon Concentration Conc. in mM g/l Sodium Gluconate 46 10.032 :
Potassium Gluconate 54 12.646 Magnesium Sulphate 5 1.232 Uncharged solutes Nannitol 105 19.13 Osmolality: 300.
________________________________________________ ________ WQ93/04578 ~ PCT/CA~2/00389 - 28 ~
T~E~ - Cont~d :.- . .- .
Name of Solution ~luco~at~ 4 :-Composition ..
Cation Anion Concentration Conc. in mM g/l ,., . ~
5sdium Gluconate 100 21.81 :.
Magnesium Sulphate 5 1.232 ~-Uncharged solutes Mannitol 105 19.13 Osmolality: 301 -.~___________________~ ______________ _________ __________ -Name of Solution Glucuronate 1 .
Composition Cation Anion Co~centration Conc. in mN g/l ,.
Sodium Glucuronate 46 10.764 Hydrogen Glucuronate 54 10.481 (Glucuronic . -Acid) .
Potassium Hydroxide 54 3.03 Magnesium Sulphate 40 ~.859 Uncharged solutes Nannitol 77 14.03 Osmolality: 301 ____________________________________________________.__~_ 2 1 1 ~ 9 1 W093/W578 PCr/CA92/00389 ~ABLE III - ~ont'd Name of 5O1ution lucurDnate 2 Composition Cation Anion Concentration Conc. in ~ g/l Sodium Glucuronate64 14.98 ~ydrogen Glucuronate76 14.74 (Glucuronic Acid) Potassium Hydroxide 76 4.26 . Magnési~m Sulphate 5 1.23 Uncharged solutes ~annitol 36 7.29 Osmolality: 301 _________________________________________________ ______.~
Nzme of Solution Gluc~om Composition Cation Anion Concentration Conc. in mN gll ~", Sodium Glucuronate92 21.54 Hydrogen Glucuronate108 20.96 (Glucuronic Acid~
Potassium Hydroxide 108 6.04 Nagnesium 5ulphate 5 1.23 Osmolality: 366 ___------W09~/04~78 ~ 3 1~ PST/CA92/00389 ABI~ Co.n~'d ~ame of Solution Glu~Prvn~e_4 -~
Composition Cation Anion Concsn~ration Conc. in ~-5Odium Glucuronate140 32.77 Nagnesium Sulphate 5 1.23 ~ncharged solutes Mannitol 36 7.29 Osmolality: 300 -~
Name of SQlution Glucuro~ate ~ :
Composition ~
Cation Anion Concentration Conc. in .
mM g/l Sodium~lucuronate 120 28.09 PotassiumHydrogen Phosphate25 3.40 -MagnesiumSulphate 5 1.23 Uncharged solutes :
Osmolality: 330 -_________________________________________________________ Name of Solution I~othionate Composition Cation Anion Concentration Conc. in ~ ~/1 :.
Sodium Isothionate 100 14.81 ~agnesium Sulphate 5 1.23 ~:
Uncharged solutes Mannitol 104 18.49 Osmolality: 306.

W093/04578 2118 917 PCT/CA9~/00389 _ 31 -~ABLEI II - Cont'd Name of Solution ~actobionat~ I
Composition Cation Anion Concentration Conc. in mN g/l Hydrogen ~actobionate46 16.48 (Lactobionic Acid) Sodium ~ydroxide 46 1.84 Hydrogen ~actobionate54 19.30 (hactobionic Acid) -Potassium Hydroxide 54 3.03 Magnesium Sulphate 40 9.85 Uncharged solutes ~annitol 72 13.13 Osmolality: 305 Name of Solution Lactobionate 2 Composition .;
Cation Anion Concentration Conc. in mM g/l ,,, Hydrogen Lactobionate 46 16.48 ~
(Lactobionic -Acid) Sodium Hydroxide 46 1.84 -~
Hydrogen Lactobionate 54 19.30 ~Lactobionic Acid) Pota~sium Hydroxide 54 3.~3 : .
Magne ium Sulphate 5 1.23 -~
Uncharged solute Mannitol 108 19.64 O~molality: 310 '.L' ______________--__________________________________________ W093/~4578 g~ 32 - PCr/CA92/00389 TABL13~ ont ' d ~ ~:

Name of Solution Lactobionate~
Composition C~tion Anion Concentration Conc~ in : -mM g/l Sodium Lactobionate 120 43.00 .
Potassium dihydrogen phosphate 25 3.40 :`
Magnesium Sulphate 5 ; 1.23 Uncharged solut~s :~
Glutathione 3 0.9212 Raffinose 30 17.83 Osmolality: 326 ~-________.________________________________________________~ ~
Name of Solution Proprio~a~e Composition Cation ~nion Concentration Conc. in `
mM g/l Hydrogen Proprionate 54 (Proprionic Acid) . .
Potassium Hydroxide 54 3.03 Sodium Proprionate 46 4.42 ~-Magnesium Sulphate 5 1.23 ~ .
Uncharg~d solutes Nannitol 108 19.64 Osmolali~y: 300 _________________________________________________________ - .

WO 93/04~78 '.~ 3 9 1rl PCr~CA~2~00389 TABI E_ ITI - Cont ~ d Name of Solution ~jaccara~e - -Composition Cation Anion Concentration Conc. in mM g/l ,, Potassium Sac~arate 85 21. 08 -Sodium Hydroxide 85 3 ~ 40 ~agnesium Sulphate 5 1. 23 ~ .
Uncharged solutes ~ . -Manni~ol 83 15.83 . ~:
Osmolality: 32 6 ; '', ;' W093J04578 ~ X9 l _ 34 _ PCI~/CA92/00389 TA8LE IV .. ;

nmB UWsolution ~ Guwso~ on #

A~a Q ~.41~0.83 1.48~0.78 * ~ ~ -8.11+1.99 3.05+1.28 * ~--120 - 12.70_4.93 3.76+1.16 *
18~ 23.97~11.27 8.03+1.13 * `

ALT~ 0 3.85_0.60 2.40~0.77 * - - -4.34~0.65 2.8~0.~0 * -120 5.08+1.30 2.77~.09 * -~ :~
18~ 7.72~2.94 ~.50~0.82 *

Bil~a 60 85~11 101+5 * -~.
120 9~14 94~6 ~-`
180 74+12 80~7 PltsC O 238+15 229+17 235+14 248~20 120 213+t2 231+18 180 216+9 240+12 # means ~ SD
* p~0.05 a l~/L/~ rNer m0/g9 1~' c X10 /L

Claims (13)

We Claim:
1. A composition for the preservation and storage of organs intended for transplantation comprising an effective amount of glucuronic acid or a physiologically tolerated salt or ester thereof in a pharmaceutically acceptable organ preservation solution.
2. The composition as claimed in claim 1, wherein the pharmaceutically acceptable organ preservation solution has substantially the composition of UW solution.
3. The composition as claimed in claim 1, wherein the pharmaceutically acceptable organ preservation solution is a UW solution modified such that it is devoid of potassium lactobionate.
4. The composition as claimed in claim 3, wherein the concentration of glucuronic acid or the physiologically tolerated salt or ester thereof is about 100 - 140 mN.
5. A composition for the preservation and storage of organs intended for transplantation, comprising a pharmaceutically acceptable preservation solution comprising:
(a) between about 3 - 200 mM of the sodium or potassium salt of glucuronate;
(b) between about 5 - 40 mM KH2PO4;
(c) between about 1 - 50 mM MgSO4;
(d) between about 5 - 50 mM raffinose;
(e) between about 1 - 50 mM adenosine;
(f) between about 0.5 - 5 mM allopurinol;
(g) between about 1 - 10 mM glutathione; and (h) between about 10 - 70 g/l hydroxyethyl starch.
6. The composition as claimed in claim 5, wherein the osmolality of the preservation solution is between about 315 and 330 mOsm/l, preferably 320 and 330 mOsm/l.
7. A method for preserving and storing an organ intended for transplantation in a patient comprising infusing said organ with a composition comprising an effective amount of glucuronic acid or a physiologically tolerated salt or ester thereof in a pharmaceutically acceptable organ preservation solution.
8. The method as claimed in claim 7, wherein the pharmaceutically acceptable organ preservation solution has substantially the composition of UW solution.
9. The method as claimed in claim 7, wherein the pharmaceutically acceptable organ preservation solution is a UW solution modified such that it is devoid of potassium lactobionate.
10. The method as claimed in claim 9, wherein the concentration of glucuronic acid or the physiological salt or ester thereof is about 100 - 140 mM.
11. A method for preserving and storing organs intended for transplantation in a patient comprising infusing said organs with a composition comprising a pharmaceutically acceptable preservation solution comprising:
(a) between about 3 - 200 mM of the sodium or potassium salt of glucuronate;
(b) between about 5 - 40 mM KH2PO4;
(c) between about 1 - 50 mN MgSO4;
(d) between about 5 - 50 mM raffinose;
(e) between about 1 - 50 mM adenosine;
(f) between about 0.5 - 5 mM allopurinol;
(g) between about 1 - 10 mM glutathione;
(h) between about 10 - 70 g/l hydroxyethyl starch.
12. The method as claimed in claim 11, wherein the osmolality of the preservation solution is between about 315 and 330 mOsm/l, preferably between about 320 and 330 mOsm/l.
13. The method as claimed in claim 11 or 12, wherein the organ intended for transplantation is the liver.
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