[CANCER RESEARCH 50. 1640-1644, March 1, 1990]

Disposition of Anticancer Drugs after Bolus Arterial Administration in aTissue-isolated Tumor Perfusion System1

Kazuhiro Ohkouchi, Hirofumi Imoto, Yoshinobu Takakura, Mitsuru Hashida, and Hitoshi Sezaki2

Department of Basic Pharmaceutics, Faculty of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606, Japan

ABSTRACT

Disposition characteristics of various anticancer drugs in a tissue-isolated tumor preparation were studied in Walker 256 carcinosarcoma-bearing rats using an in situ single-pass vascular perfusion technique.Three anticancer drugs, 5-fluorouracil, mitomycin C, and Adriamycin,and two lipophilic prodrugs of mitomycin C were tested in the tumorpreparation perfused with Tyrode's solution containing 4.7% bovine

serum albumin. After bolus arterial injection of test drugs, their outflowconcentration-time curves were analyzed based on statistical momenttheory. In each tumor preparation, the injection of drug was paired withthat of vascular reference substance, Evans' blue-labeled bovine serum

albumin, and disposition parameters of the drug were corrected withthose of vascular reference substance. From the mean transit time valuesof vascular reference substance, the average vascular volume of the tumorpreparation was calculated to be 0.063 ml/g, which decreased with tumorgrowth. All drugs showed significant extraction by the tumor tissue,depending on their physicochemical properties. Distribution volumes oftested drugs were from 1.53 to 3.33 times larger than the vascular volume.Calculated intrinsic clearance values for the protein-unbound fractionsincreased as the lipophilicity of the drug increased. The potential increasein tumor uptake was observed in lipophilic prodrugs of mitomycin C. Thepresent experimental system is thus suggested to be useful for analyzingdrug disposition in tumor tissue.

INTRODUCTION

In cancer chemotherapy, it is necessary to concentrate cyto-toxicity of an anticancer agent to the tumor and to minimizethe exposure of other normal tissues to it. To accomplish this,we have reported several efforts, such as alternation of the routeof administration or dosing schedule and physical or chemicaltransformation of drugs. In our series of investigations we havedeveloped lipophilic and polymeric prodrugs of MMC' and

examined their physicochemical, pharmacodynamic, and phar-macokinetic properties (1,2). These studies demonstrated thatthe in vivo disposition of MMC could be controlled by choosingphysicochemical characteristics of carrier moiety in prodrugs.

The intraarterial administration of anticancer drugs includingthe chemoembolization approach has become a matter of greatinterest (3, 4). However, little information has been obtainedon disposition profiles of anticancer drugs from the intravas-cular space to tumor tissues. In a series of investigations, wehave established an experimental system for analyzing drugdisposition in a local tissue based on the statistical momenttheory (5). The disposition of various drugs in the rat liver andin the normal and tumor-bearing muscles of the rabbit (6, 7)has been studied with this system, and its relation with thephysicochemical properties of drugs was elucidated.

In the present study, disposition characteristics of anticancer

Received 5/23/89; revised 10/23/89; accepted 11/27/89.The costs of publication of this article were defrayed in part by the payment

of page charges. This article must therefore be hereby marked advertisement inaccordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1Supported in part by a Grant-in-Aid for Scientific Research from the Ministryof Education, Science, and Culture, Japan.

2To whom requests for reprints should be addressed.3The abbreviations used are: MMC. mitomycin C; ADR, Adriamycin; FU, 5-

fluorouracil; propyl-MMC, propyloxycarbonyl-mitomycin C: pentyl-MMC, pen-tyloxycarbonyl-mitomycin C: BSA. bovine serum albumin: VRS, vascular reference substance; EB/BSA, bovine serum albumin labeled with Evans' blue.

drugs and prodrugs were studied in a tissue-isolated Walker256 tumor preparation in the rat.

MATERIALS AND METHODS

Chemicals. MMC, ADR, and FU were kindly supplied by KyowaHakko Kogyo Co., Tokyo, Japan. Propyl-MMC and pentyl-MMC weresynthesized as reported previously (8-10). S-Fluoro[2-'4C]uracil was

purchased from CEA, Cedex, France. All other chemicals were ofreagent grade and obtained commercially.

Animals and Tumors. SLC female Wistar rats weighing 100 to 120g were obtained from Shizuoka Agricultural Association for LaboratoryAnimals, Shizuoka, Japan. Walker 256 sarcoma was kindly suppliedby Shionogi & Co., Osaka, Japan, and maintained by s.c. inoculationto another rat every 2 wk.

Preparation of Tissue-isolated Tumors. Ovarian tissue-isolated tumors were prepared according to the method of Cullino and Grantham(11). Rats were anesthetized by i.p. injection of pentobarbital. Theovary, uterus, and mesovarium were pulled from the peritoneal cavitythrough a 1-cm incision in the skin and muscle wall of the left lumbarregion. The uterine horn was ligated distally to the origin of the ovary,and the mesovarium and ovary were ligated proximally to the ovarianartery and vein. Three blocks of minced Walker 256 sarcoma wereinoculated in the adipose tissue around the ovary, and the inoculatedadipose tissue was wrapped with Sealon film (Fuji Film Co., Tokyo,Japan) to separate from other tissue. The Sealon film sheet was folded,and its margins were melted to form a sealed envelope. The tumor-inoculated tissue was placed between the cutaneous and muscle wall inthe abdomen. Streptomycin sulfate (Sigma, St. Louis, MO) and apenicillin G potassium (Toyo Jozo Co., Tokyo, Japan) were administered topically before closing the skin. After 1wk, the film was changed.Two wk after tumor inoculation, the tumor that had grown to 2 cm indiameter and 8 g in weight was used for the perfusion experiment.

Perfusion Experiment. The tumor was perfused as shown in Fig. 1.To achieve independent perfusion for the isolated tumor, all bloodvessels supplying nontumorous tissues (left renal artery and vein, rightrenal vein, inferior vena cava, and aorta) were ligated. The inferior venacava and aorta were cannulated with vinyl tubes and perfused at therate of 0.8 ml/min. No leakage of perfusate (EB/BSA) was observed inthe arterial side after these procedures. It took 15 min to purge thesystem of blood and to allow for equilibration between tumor andperfusate. The drug was introduced from the arterial side using a six-position rotary valve injector as a pulse function. The tumor mass wasperfused with Tyrode's solution (a mixture of 137 mM NaCl, 2.68 mM

KC1, 1.80 mM CaCl2, 11.9 mM NaHCO3, 0.362 mM NaH2PO4, 0.492mM MgCl2, and 5.55 mM D-glucose) containing BSA (Fraction V) at aconcentration of 4.7% (w/v). The same medium was used for druginjection. The concentrations of injection solution (0.1 ml) were 10mg/ml for FU (1 ^Ci/ini for 5-fluoro[2-14C]uracil; 1 mg/ml for EB,MMC, and pentyl-MMC; and 0.1 mg/ml for propyl-MMC. The perfusate was gassed with 95% O2-5% COi and maintained at 37°C.Evans'

blue/BSA was used as a VRS and applied to the tumor preparationprior to the drug injection for obtaining basic information about eachtumor tested.

Sampling and Assay. The outflowing perfusate was collected intopreviously tared tubes at appropriate time intervals (at first 5 s, subsequently 10 to 15s). The sample volume was calculated from the gainin weight of the tube by assuming a density of the outflow perfusate lobe 1.0. The sample was subjected to assay after an appropriate dilutionwith the perfusate buffer. The concentration was determined speclro-photometrically at 620 nm for EB/BSA and at 479 nm for AI>K.

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DISPOSITION OF ANTICANCER DRUGS

Rat

Perfusote

Fig. 1. Perfusion system of tissue-isolated tumor preparation.

Table 1 Derivation of the disposition parameters from momentsQi, auc, vu*, and »,.VRSare mean inflow rate, auc, of VRS. and r, of VRS,

respectively.

ruminationExtent

Amount of recoveryRecovery ratio

RateMean elimination time

ClearanceIntrinsic clearance

DistributionExtent

Distribution volumeTissue distribution ratio

A, = auc,-0,F, = auc,/auc,.vRs

<.,.i= t,/E,

CL,„.,= K/fcw

MMC, propyl-MMC, and pentyl-MMC were assayed by a high-performance liquid chromatography system (Trirotar; Jasco, Tokyo, Japan) equipped with a variable wavelength UV detector (UVIDEC 100-II; Jasco) at 365 nm. The stationary phase used was a Cosmosil 5C|8-packed column (4.6 x 150 mm; Nacalai Tesque, Kyoto, Japan), and ashort column packed with Lichrosorb RP-2 (E. Merck, AG, Darmstadt,West Germany) was used to guard the main column. Methanohwatermixtures (35:65 for MMC and 60:40 for propyl-MMC and pentyl-MMC) were used as the mobile phase with a flow rate of 0.8 ml/min.The precise limit of sensitivity was 0.1% of dose/ml. For the determination of FU, 5 ml of scintillation medium (Cleasol; Nacalai Tesque,Kyoto, Japan) were added to the sample, and I4C radioactivity was

measured in a liquid scintillation system (LSC 900; Aloka, Tokyo,Japan).

Pharmacokinetic Analysis of Outflow Pattern. Data analysis wascarried out based on the statistical moment concept. The detailedtheoretical description of this analysis was given previously (1,7). TheOth and first moment parameters (moments) for output response aredefined as follows

auc, = Cdt

' x

where t is time and C is the concentration of a test substance normalizedby injection dose with a dimension of "% of dose/ml." auc, and f, are

the area under the concentration-time curve and the mean transit time,respectively. The moments were calculated by numeral integration usinga linear trapezoidal formula from the outflow concentration-time curve.All f, values were corrected by subtracting the time corresponding tothe transit through afferent and efferent tubes.

In the next step, distribution parameters were calculated from themoments as summarized in Table 1 (5). Each parameter is representedas: RI, recovery percentage of drug corrected by outflow flow; /•"„

recovery ratio of drug to that of VRS; tríj,mean time necessary forelimination of the drug from the perfusate compartment; CLim.,,intrinsic clearance; Vt,distribution volume; and k,, ratio of amount of drugin the extravascular tissue to that in the perfusate.

Determination of Partition Coefficient of Drug. The partition coefficient of the drug was determined as a ratio of the drug concentrationbetween 20 ml of octanol and water after mixing and equilibration at37°Cfor 48 h.

Determination of Protein Binding. The nonbinding percentage of drugwith BSA in the perfusate (fu) was determined by the centrifugalultrafiltration method (Ultracent-30; Tosoh Co., Tokyo, Japan). Theintrinsic clearance for the free fraction of drug (CLin,.fill)was calculatedas CU,.,.. = CLinl.,/fu.

Angiography of Tumor Tissue. Megiumine diatrizoate (Sigma) wassolvated in the perfusate as 65% (w/v) and continuously injected fromthe artery during soft (low kVp) X-ray examination.

Determination of True Vascular Volume. The vascular volume of thepresent preparation calculated as the V¡value of VRS includes not onlyvascular space in the tumor mass but also that of appendicular vascularmatter between the tumor mass and the cannulation points. The latterwas determined from the residual perfusate volume in there at the endof the perfusion experiment and subtracted from the V¡value of VRSto estimate the true vascular volume of the tumor.

RESULTS

Tumor Perfusion. An angiogram of the tumor preparationconfirmed the establishment of the isolated perfusion systemfor the Walker 256 tumor mass (Fig. 2). The mean recovery ofthe perfusate from the venous cannula was about 58.1% in thissystem.

Outflow Patterns in the Tumor Tissue. Fig. 3 shows the typicalconcentration-time curves of FU (A) and pentyl-MMC (B),together with those of VRS in the same tumor preparations.

Fig. 2. Angiogram of tissue-isolated tumor preparation, t, tumor tissue: a,arterial inflow side; v, venous outflow side.

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DISPOSITION OF ANTICANCER DRUGS

100 200

Time(sec)100 200

Time(sec)

Fig. 3. Typical outflow patterns of FU and pentyl-MMC together with EB/BSA. The solid lines drawn through the points were obtained by eyeball fit. Left,O. EB/BSA: A. FU. Right, O, EB/BSA; •¿�pentyl-MMC.

Table 2 Moment parameters for test drugs and l 'RS in single-pass perfusion

experiments with the tissue-isolated tumor preparationStatistical significance (Fisher's pairing t test. P< 0.05) for auc, was recognized

between the groups (FU, MMC. propyl-MMC, pentyl-MMC < each EB/BSA)and (propyl-MMC, pentyl-MMC < FU). Statistical significance (Fisher's pairing

t test, P < 0.05) for r, was recognized between the groups (FU > EB/BSA. MMC.ADR. propyl-MMC. pentyl-MMC).

CompoundEB/BSA

FUEB/BSA

MMCEB/BSA

ADREB/BSAPropyl-MMCEB/BSA

Pentyl-MMCauc,

(% of dose •¿�s/ml)7391

±122°

6290 ±4738430

±3885678 ±4076959

±3615668 ±10838163

±5994797 ±16008024

±7404526 ±580Ms)43.5

±15.9117.8±32.456.6

±21.972.1 ±18.637.4

±9.842.3 ±6.450.3

±18.450.4 ±14.259.9

±30.068.6 ±22.6Tumor

wt(g)5.92

±2.148.04

±1.317.57

±4.709.28

±3.758.69

±4.65(«)(5)*(3)(3)(5)(3)

' Mean ±SD.* Numbers in parentheses, experimental number.

The concentration of each test drug was lower in the initialphase and higher in the later phase than those of VRS. Especially, the FU concentration showed a gentle decline in theperiod from 100 s after injection when it overcame that of VRS.Outflow patterns of ADR, MMC, and propyl-MMC were basically similar to that of pentyl-MMC (data not shown).

Moments and Disposition Parameters. Tables 2 and 3 summarize the moments and the disposition parameters obtainedin this system, respectively. All test compounds showed smallerauc, and longer /, than those of VRS, which were obtained bypreceding injection in the same tumor preparation (Table 2).Disposition parameters representing distribution, elimination,and clearance of drugs varied depending on their physicochem-ical properties. The mean value of K,of VRS which correspondsto the apparent vascular volume of the tumor tissue preparationwas 0.087 ml/g of tissue in animals listed in Table 3. The V¡ofFU was more than 3 times as large as that of VRS. A, calculatedas the product of flow and auc, normalized by the dose variedin the range between 57.8% for pentyl-MMC and 82.9% forFU, while that of VRS was almost 100%. We calculated therecovery ratio of each drug (F¡)by assuming that of VRS to be1.0.

Vascular Volume of Tumor. Fig. 4 shows the relation betweenthe vascular volume and the weight of all tested tumor prepa

rations. The vascular volume of the tumor decreased with anincrease in tumor weight.

Effect of Protein Binding of Drugs on Intrinsic Clearance.Table 4 summarizes the intrinsic clearance values of test compounds corrected for protein-unbound fraction (CLim,,.u) together with their physicochemical properties. These resultsshow that the value of CLim.,.„increased as the lipophilicity ofdrugs increased.

DISCUSSION

In general, drugs are rapidly extracted from the blood byseveral organs such as the liver and kidney, and these conditionsare considered to be disadvantageous for systemic drug deliveryto the tumor tissue in cancer chemotherapy. Therefore, localadministration modalities such as intraarterial infusion areadopted to enhance the drug exposure to tumor tissue. Toimprove the efficiency of intraarterial infusion, several approaches have been reported such as vascular obstruction, useof vasoactive agents (12, 13), utilization of oily medium (14),and chemoembolization using microcapsules (15). Chemicalmodification of drugs into a lipophilic or a polymeric form alsowould be a promising strategy.

The purpose of this study was to estimate the pharmacoki-netic behavior of anticancer drugs and their prodrugs in a tissue-isolated tumor preparation using an analytical method for localdrug disposition based on moment theory. A tissue-isolatedpreparation has been applied to elucidate physiological properties of tumors such as blood flow, vascular volume, andoxygen and glucose consumption (16), and more recently, tostudy the effect of hyperglycemia on hemodynamics and pHresponses in the tumor (17). In spite of these basic researchinvestigations, however, few studies have been done on drugdisposition using this method. In a previous paper, we haveinvestigated the disposition of MMC and its polymeric pro-drugs in the single-pass perfusion system of the tumor-bearingmuscle of rabbits (2). From this study, however, only mixedinformation for both the normal and tumor tissues was obtained. In the present study, disposition characteristics of drugsin the tumor tissue can be independently analyzed in a tissue-isolated tumor preparation. Using this tissue-isolated tumorpreparation, we can manipulate some experimental conditions,such as perfusate composition, and appreciate the effects ofperfusion rate, injection medium, hyperthermia, and so on.

In the present study, drugs and prodrugs were introduced asa pulse (7.5 s) function from the arterial side, and venousoutflow patterns were analyzed by statistical moment theory.Although tumor tissue was shown to be perfused and wasisolated from other tissues by the present experimental procedure, as shown in Fig. 2, the mean outflow recovery was onlyabout 58.1%. In contrast to the liver and muscle perfusionexperiments which gave almost 100% outflow recovery (2, 6),it was difficult to satisfactorily recover the flow from the venousside in this tumor perfusion system, mainly due to leakage fromthe unligatable venous vessels. In the present analysis, however,the disposition parameters can be estimated in spite of lowoutflow recovery, because when plural processes exist in aperfusion circuit after the passage through the tumor, theoutflow concentration should be the same in each pathway.Assuming that all of perfusate passed through the tumor tissueand then loss of perfusion fluid occurred after this, the disposition parameters could be calculated from the outflow concentration-time curve and inflow rate (Q). The fact that there wasno permeate in the arterial side in Fig. 2 and other experiments

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Table 3 Amount of recovery and disposition parameters for test drugs and YRS in single-pass perfusion experiments with the tissue-isolated tumor preparationStatistical significance (Fisher's pairing t test, P < 0.05) for A, was recognized between the groups (FU, MMC, ADR, propyl-MMC', pentyl-MMC < each EB/

BSA), (FU, ADR > propyl-MMC), and (FU, ADR > pentyl-MMC). Statistical significance (Fisher's pairing / test, P < 0.05) for F¡was recognized between the groups(FU, MMC. ADR, propyl-MMC. pentyl-MMC < each EB/BSA). (FU > MMC, propyl-MMC, pentyl-MMC), and (ADR > pentyl-MMC). Statistical significance(Fisher's pairing r test, P < 0.05) for t«uwas recognized between the groups (FU > MMC, ADR. propyl-MMC). Statistical significance (Fisher's pairing t test, P <0.05) for CLtau was recognized between the groups (FU > propyl-MMC). Statistical significance (Fisher's pairing t test. P < 0.05) for \\ was recognized between thegroups (FU > EB/BSA. MMC, ADR, propyl-MMC, pentyl-MMC). Statistical significance (Fisher's pairing I test. P < 0.05) for k, was recognized between the groups(FU > MMC, ADR, propyl-MMC, pentyl-MMC).

EB/BSAFUEB/BSA

MMCEB/BSA

ADREB/BSA

Propyl-MMCEB/BSA

Pentyl-MMCK,

(%)97.5±2.9°

82.9 ±6.0106.5

±6.870.4 ±4.995.8

±5.278.1 ±15.1105.3

±9.561.7 ±19.6102.6

±8.957.8 ±8.1F,1

0.851±0.0540.664

±0.0841

0.812 ±0.1181

0.587 ±0.1821

0.572 ±0.130>„.:

(S)827

±184230

±106308

±221154

±100160

±7.8CU,.,

(ml/min/g)0.0247

±0.00910.0484

+0.01200.0255

±0.00430.0875

±0.07690.0744

±0.0095V>

(ml/g)0.0987

±0.02190.329 ±0.0900.0917

±0.04470.1 72±0.0370.0792

±0.03210.121±0.0710.0949

±0.0850.175±0.1570.0923

±0.0140.1 97 ±0.020*,0

2.34 ±0.660

1.06 ±0.550

0.44 ±0.290

0.85 ±0.400

1.01 ±0.11°Mean ±SD.

Timor Height (g)

Fig. 4. Relation between tumor weight and intravascular volume. The vascularvolume was shown to be represented against tumor weight by the followingequation in weight area between 1 and 20 g: Intravascular volume (ml/g) =-0.00494 x tumor weight (g) + 0.107.

Table 4 Physicochemical properties and intrinsic clearance corrected for protein-unbound fraction of test drugs

CLtmjj, was calculated as CL|„t,,/fu.Statistical significance (Fisher's pairing t

test, P< 0.05) was recognized between the groups (FU < propyl-MMC) and (FU,MMC, ADR, propyl-MMC < pentyl-MMC).

FUMMCADRPropyl-MMCPentyl-MMCM,130

334544420

488PC,»"0.152

0.3600.463

32.7279fu0.9813

0.89350.49150.6910.160CLín,.,..

(ml/min/g)0.0252

0.05410.0518

0.1270.4650.0093"

0.01340.00870.1110.060

°PC»,,,partition coefficient of drug between octanol and water: fu. nonbinding

percentage of drug with BSA.* Mean ±SD.

with dye (EB/BSA) perfusion support these assumptions. Thevalidity of this analysis was also confirmed by the result thatthe amount of recovery (R,) of VRS was almost 100% in allgroups (Table 3).

Tumor tissues are characterized by enhanced vascular permeability to macromolecules (18, 19). However, extravasation ofEB/BSA would be negligible during a single passage of tumortissue; tumor uptake rates of BSA were calculated to be 30 n\/h/g for rabbit VX2 carcinoma and 14.2 Ml/h/g for mouse S-180 carcinosarcoma (20) in previous studies. Therefore, the V¡of EB/BSA can be considered to correspond to the vascularvolume of tumor tissue. We have already obtained reasonable

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values for the vascular volume of rabbit muscle (5) and rat liver(21) by the same procedure.

The vascular volume of the present tissue-isolated Walker256 carcinoma in the weight range of 1 to 20 g was shown tobe represented, versus total tumor weight, by the equation,vascular volume (ml/g) = —¿�0.00494x tumor weight (g) +

0.107. That is the vascular volume content decreased with anincrease in tumor weight, and the vascular volume content ofan average size tumor (8.71 g) was 0.070 ml/g (Fig. 4). Songand Levitt (22) reported that the average vascular volume ofs.c. implanted Walker 256 carcinoma was 0.079 ml/g and thatthis volume decreased linearly with tumor size, using 5lCr-

labeled erythrocytes. Cullino and Grantham (23) found that thevascular volume of tissue-isolated Walker 256 carcinoma determined with Dextran 500 (M, 375,000) was almost constant atthe value of about 0.1 ml/g when the tumors were small (4 to12 g), but no correlation was observed in larger tumors. Thepresent results are essentially in good agreement with thesedata.

In this study, disposition parameters for drugs representingdistribution, elimination, and intrinsic clearance were calculated from the primary moment parameters (Table 3). Tocompensate for the variance among tumor preparations, parameters for each drug were determined together with those of VRSin the same preparation.

FU showed peculiar disposition characteristics among thetested compounds, i.e., a large V{,low extraction (large /•",-),and

large teU. These results suggest that FU easily moves to theextravascular space during the passage of drug solution, butmost of it returned to the intravascular space in the followingstep. In MMC and ADR, 34 and 19% of administered dosewere taken up by the tumor, respectively, suggesting theireffective delivery to the tumor by intraarterial administration.Two types of lipophilic prodrugsof MMC showed a little largerV¡and smaller F¡than the parent drug, suggesting an enhanceduptake by the tumor. It was difficult, however, to discuss thedisposition characteristics of lipophilic prodrugs from thesedata, since the effect of protein binding cannot be neglected.

In order to examine true disposition characteristics, intrinsicclearance corrected by the free fraction (CLinl, „¿�)of each compound was calculated (Table 4). The correlation between lipo-philicity and protein binding indicates that the lipophilic drugsare likely to bind albumin (24). It was clear that the CL¡n,,,,uincreased with an increase in lipophilicity and that pentyl-MMC

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DISPOSITION OF ANTICANCER DRUGS

had the highest CLin,.,,„value. This indicates that chemicaltransformation of a drug into a lipophilic form is desirable forthe delivery- to the tumor from the vascular space. However, the

difference in the net uptake of the drug by the tumor (£,)wasnot so large, since the lipophilicity also increased the protein-bound fraction which basically did not contribute to the tumoruptake. It is concluded from these results that intraarterialinfusion of anticancer drugs especially lipophilic prodrugswould show higher uptake by the tumor when applied in abinder (albumin)-free formulation.

Prodrugs must be regenerated to the parent drug beforeexhibiting therapeutic activities. Regeneration processes of thepresent M MC prodrugs have been analyzed previously in therabbit muscle through Laplace transformation of the convolution function for them (7). It was confirmed that lipophilicderivation produced a larger tissue distribution and that rapidregeneration of the drug from the prodrug resulted in a largelocal bioavailability of the parent drug.

Thus, the utility of a novel experimental system for analyzingthe drug disposition in a tissue-isolated tumor preparationbased on the statistical moment concept was demonstrated. Thepresent system also can be applied for the evaluation of variousdrug delivery approaches, such as utilization of a macromolec-ular or spherical (liposome, microsphere) carrier system andcombination chemotherapy with vasoactive agents and hyper-thermia, etc. Especially, macromolecular carrier systems seemto be of interest since macromolecules have some advantageousfeatures for tumor targeting (25).

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1990;50:1640-1644. Cancer Res   Kazuhiro Ohkouchi, Hirofumi Imoto, Yoshinobu Takakura, et al.   Administration in a Tissue-isolated Tumor Perfusion SystemDisposition of Anticancer Drugs after Bolus Arterial

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