Cisplatin (CDDP) is an antitumor agent against several types of neoplasms. It has, however, various side effects such as nephrotoxicity and ototoxicity. Several efforts have been made to prevent these toxic side effects. On the other hand, deferoxiamine mesilate (DFO), an iron-chelating agent, has been used for iron-overloaded patients. Since the 1980s, many authors have reported DFO-associated hearing impairment. Some experimental data suggest that DFO itself was responsible for ototoxicity. In addition, it has also been recognized that DFO can act as a free radical scavenger. Experimental trials using DFO are also expected to prevent CDDP-induced toxicity because its generation mechanism is thought to be associated with free radical formation. The present study was planned to investigate whether DFO, which might be an ototoxic agent, had a protective effect against various CDDP-induced toxicities including ototoxicity. Fisher rats were used in this study and were divided into four groups: 1) Group I, a vehicle control, 2) Group II, animals receiving 100 mg of DFO per kg, 3) Group III, animals administered 0.9 mg of CDDP per kg alone and 4) Group IV, animals receiving 100 mg of DFO per kg 60 min before 0.9 mg of CDDP per kg for 10 days. First, the protective effect of DFO against CDDP-induced ototoxicity was studied. The auditory threshold was determined by using the compound action potential (CAP) from the round window membrane. CAPs were recorded on the 5th day after completion of drug administration. Then CAPs recording, cochlear sensory epithelia were observed over all the turns of the cochlea by scanning electron microscopy. There were no significant differences in CAP thresholds between Group I and IV, though the thresholds in Group III were significantly higher at 16 kHz and 20 kHz than those in Group IV. The rate of missing outer hair cells in Group IV was significantly lower than that in Group III. The results clearly demonstrated that DFO had a protective effect against CDDP-induced ototoxicity. Second, the protective effect of DFO against CDDP-induced nephrotoxicity was studied. Renal function was evaluated by measuring blood urea nitrogen (BUN) and serum creatinine (Cr) levels. Both BUN and Cr levels in Group IV were significantly lower than those in Group III. The data suggested that DFO preadministration prevented CDDP-induced nephrotoxicity. Third, the influence of DFO on the antitumor activity of CDDP was investigated in rats inoculated with squamous cell carcinoma cells (SCC-131) subcutaneously. The influence of drugs was determined by measuring the tumor growth rate. There was no difference in the tumor growth rate between Group III and IV. The result revealed that DFO had no influence on CDDP antitumor activity. In conclusion, the above results demonstrating that DFO preadministration can prevent both CDDP-induced ototoxicity and nephrotoxicity without attenuation of CDDP antitumor activity, suggest the usefulness of CDDP antitumor chemotherapy.

To evaluate the possible clinical intrathecal use of 5-fluoro-2'-deoxyuridine (FdUrd) for malignant brain tumors, its anti-tumor activity and neurotoxicity were compared with that of 5-fluorouracil (5-FU) and 5-fluorouridine (FUrd) in vitro. FdUrd showed good tumoricidal activity against cultured mouse 203 glioma cells and rat Walker 256 carcinoma cells as well as A172 human glioblastoma cells. Daoy human medulloblastoma cells and CADO-LC4 human lung cancer cells. It also showed less toxicity for primary cultures of neurons from C57/BL6 mouse and human embryo compared to 5-FU and FUrd. Thymidine phosphorylase (TPase) and thymidine kinase (TK), key enzymes for metabolism of 5-FU derivatives, were measured in cerebrospinal fluid (CSF). TPase or TK activity was detected in the CSF of hardly any patients with malignant brain tumors including meningeal carcinomatosis. These data indicated that the CSF is a favorable site for FdUrd chemotherapy, because the rate of conversion of FdUrd injected to 5-FU would be minimal. In conclusion, FdUrd may be potentially useful for intrathecal treatment of meningeal carcinomatosis.

The present study was designed to investigate the appropriate intravesical retention time of pirarubicin (THP) for the treatment of bladder tumor in terms of its anti-tumor effect and side effect. We administered THP to 22 patients with superficial bladder tumor intravesically and measured the THP concentration in the tumor tissues. Patients were divided into 3 groups by retention time; 8 for 30 minutes, 8 for 1 hour and 7 for 2 hours. Tumor tissues were obtained by transurethral resection 30 min., 1 or 2 hours after the intravesical instillation of THP at the fixed concentrations of 30 mg/30 ml. There was no significant difference in THP concentration between 3 groups. This indicates that the anti-tumor effect of intravesical instillation of the THP would be expected by only 30 min. of intravesical retention time at the THP concentration of 30 mg/30 ml. Then, we administered 30 mg/30 ml of THP solution for 30 min. to 10 patients intravesically 6 times every 48 hours to investigate its clinical anti-tumor effect and side effect. There were 2 complete responses, 3 partial responses and 5 cases with no changes for a total response rate of 50%. No side effect was observed. It is considered that 30 min. of intravesical retention time at the THP concentration of 30 mg/30 ml would be appropriate in terms of its anti-tumor effect and side effect.

A 10-center cooperative clinical study with a new formulation of epirubicin hydrochloride injectable solution (Epirubicin-RTU) was conducted in patients with hepatocellular carcinoma. Epirubicin-RTU 60 mg/m2 was injected into the hepatic artery and a three-week drug-free interval followed. Of 15 patients with hepatocellular carcinoma registered in this study, 14 patients were eligible, and they all completed the entire course. The objective was to investigate the safety of treatment with Epirubicin-RTU in 14 eligible patients. The adverse drug reactions frequently observed in these 14 eligible cases were leukopenia, neutropenia, thrombocytopenia, alopecia, and fever. They were all reversible and tolerable. With these results. Epirubicin-RTU was considered to be a safe pharmaceutical product to inject into the hepatic artery.

We analyzed PyNPase expression discriminating between cancer and tumor stroma of the colorectum by Western blotting using a newly developed extraction method from microdissected tissue sections fixed with buffered formalin. Analysis of 98 colorectal cancers revealed that PyNPase was as high as 70.2 +/- 18.5 unit/mg protein in the stroma fraction (SF), whereas it was 45.1 +/- 10.5 in the cancer fraction (CF) (p < 0.0001). Vessel density was correlated with PyNPase in the SF but not in the CF. In stage IIIb, 11 cases expressing a high level of PyNPase in the CF showed poorer prognosis than 10 cases with low-level PyNPase expression (p < 0.05), although the level of PyNPase expression in the SF did not affected the patients prognosis. Immunohistochemical examination indicated that PyNPase in the SF was mainly produced by macrophages (M phi), and therefore we investigated the profile of PyNPase production by M phi. In in vitro experiments PyNPase production by M phi was greatly enhanced by stimulation with OK-432, and the culture supernatant had the ability to convert 5'DFUR to 5-FU.

We present a case of asynchronous development of transitional cell carcinoma in urinary bladder and renal pelvis after prolonged cyclophosphamide therapy. A 57-year-old woman had received 290 g cyclophosphamide for 13 years because of therapy for non-Hodgkin lymphoma. She was suffered from dysuria and macrohematuria and visited our clinic. Cystoscopy, CT and MRI revealed invasive bladder tumor and total cystectomy was performed. Histological diagnosis was transitional cell carcinoma, G3 < G2, pT4. Six months after the cystectomy, a follow up urography and computerized tomography showed left renal pelvic tumor. The patient underwent total nephroureterectomy, and the histological diagnosis was transitional cell carcinoma, G3, pT3. We reviewed cyclophosphamide induced urothelial carcinomas from Japanese and world literatures.

123I-metaiodobenzylguanidine (MIBG) myocardial single photon emission computed tomography (SPECT), 123I-beta-methyliodophenyl pentadecanoic acid (BMIPP) myocardial SPECT, and holter ECG recording were performed in patients with non-Hodgkin's lymphoma who underwent chemotherapy including pirarubicin (THP), in an attempt at early detection of cardiac toxicity from THP. Twenty-six patients with untreated non-Hodgkin's lymphoma who received THP-COPBLM therapy were studied. For THP-COPBLM therapy. THP was administered at a dose of 40 mg/m2 every 21 days and the total dose was 250 mg/m2 on average (40 approximately 400 mg/m2). 1) The washout rate (WR) correlated with the total THP dose, and was considered to be a useful index of cardiac sympathetic nervous dysfunction. 2) The left ventricular ejection fraction (LVEF) correlated negatively with the total dose of THP. 3) The total dose of THP showed a correlated positively with the extent score and severity score determined by BMIPP. 4) The WR correlated with the frequency of premature ventricular contraction. Animal studies have indicated that THP has less cardiac toxicity than doxorubicin, but the present study showed that cardiac toxicity occurred at a total THP dose of about 360 mg/m2 in elderly patients. Accordingly, when THP is used to treat elderly patients, multimodal evaluation of cardiac is necessary to detect cardiotoxicity and to determine the optimal dosage.

A human colon cancer cell line, HCT-15, was found to proliferate in human sera. Sera of cancer patients undergoing continuous intravenous administration of 5-fluorouracil (5-FU) were tested for the antitumor effect by counting HCT-15 cells stained with Giemsa solution. 5-FU concentrations in those sera were not correlated with the antitumor effects. After HCT-15 cells were incubated in sera of cancer patients or healthy volunteers for 24 hours followed by removal of those sera, those cells were incubated in a culture medium containing 500 or 1000 ng/ml of 5-FU for 48 hours. It was shown that preincubation of HCT-15 cells with sera of cancer patients but not sera of healthy volunteers decreased the sensitivity of HCT-15 cells to 5-FU. These results suggest that there may be some factors in the serum of a cancer patient, which affect the antitumor effect of 5-FU.

Toremifene (TOR) is a new antiestrogenic agent, a triphenylethylene derivative that was developed as an alternative to tamoxifene (TAM). TOR has been observed to be more effective than TAM with milder toxicity at high doses. We examined the in vitro combination-effect of TOR with cyclophosphamide, adriamycin, 5-fluorouracil and three drug mixture (CAF) on the growth of various human mammary carcinomas. The combination shows a semi-additive or additive growth inhibitory effect on all estrogen positive cells used here except one cell line. In particular, the additive or synergic combination-effect was observed on TAM resistant cells. Furthermore, TOR exhibits a chemosensitizing activity in ADR-resistant cells by expressing P-glycoprotein coded by MDR-1 (multidrug resistance gene). The chemosensitizing activity is dose-dependent of TOR. As described above, the combination of TOR with CAF shows more than a semi-additive effect in this experiment. In conclusion, the addition of high-dose TOR to CAF therapy might be useful for advanced/recurrent breast cancer.

This paper describes the antitumor effect of human lactoferrin against the human pancreatic cancer cell line SPA, which was newly established in our laboratory from a metastatic liver tumor of pancreatic origin. In tissue culture, the cancer cells proliferated rapidly at 16 hours of doubling time, and produced tumor markers into the culture medium at a high concentration. Subcutaneous and intraperitoneal transplantation of these neoplastic cells into nude mice resulted in tumor formation and carcinomatous peritonitis. The inhibitory effect of human lactoferrin on the cell growth of SPA was found both in vivo and in vitro. There was significant inhibition of cell growth in vivo at the concentration of 1 microgram/ml of hLf in the culture medium. And in the in vivo assay, hLf delayed the growth of subcutaneously transplanted tumors into BALB/c nude mice, and the effect was retained for two weeks. These results indicate the possibility that hLf will become one of the new drugs for adjuvant therapy against pancreatic cancer.

We measured mitomycin C (MMC) concentrations in rabbit ocular tissues, using high-performance liquid chromatography, after the subconjunctival application of surgical sponges which had absorbed various doses (0.1, 0.2 or 0.4 mg) of MMC and with varying duration of application (1, 3, or 5 minutes). MMC concentrations at the administered site showed a significant correlation with the dose and duration of application. In multiple regression analysis the MMC concentration (microgram/g) in the conjunctiva was -6.73 + 67.4 x dose (mg) + 1.66 x time (minutes) (R-square 0.65); in the sclera, -1.85 + 38.2 x dose + 0.927 x time (R-square 0.63); and in the corena, -0.727 + 8.44 x dose (R-square 0.46). We also compared the results of filtering surgery on rabbits using 0.2 mg MMC intraoperative exposure of 1, 3, 5 minutes or 5-minute application of distilled water (control). The duration of maintained intraocular pressure reduction and filtering bleb was significantly longer in MMC-treated group (1 minute; 15 +/- 7.1, 3 minutes; 27 +/- 14.3, 5 minutes; 29 +/- 18.6 days, mean +/- standard deviation) than in the control group (7 +/- 3.7 days). Among the MMC-treated groups, good results were maintained significantly longer in the 3- or 5-minute application groups than in the 1-minute group. There was no difference between the 3-minute group and the 5-minute group.

Accumulated evidence indicates that drug resistance is induced in tumor cells treated with a variety of anti-cancer drugs and that there is a possibility of cross-resistance to ionizing radiation associated with induced drug resistance. Most in vitro studies have shown inconsistent results on cross-resistance probably because of different cell lines used and protocols for drug induction. In this study, TE3 human squamous cell carcinoma cell line was treated with a 4-day cycle of cisplatin (cis-diamminedichloroplatinum (II); CDDP) at a concentration yielding 10% cell survival. The treatment was repeated up to 3 cycles. After treatment, cells were tested for CDDP and X-ray sensitivity. One cycle of CDDP treatment induced CDDP resistance with a factor of 1.41 and 2 cycles of the treatment with a factor of 1.86. The resistance factor reached a plateau at 3 cycles of treatment. For analyzing the correlation of CDDP and X-ray resistance, 30 clones from both untreated and 3-cycle treated cells were isolated and analyzed for CDDP and X-ray sensitivity. The sensitivity was expressed as the concentration of drug or dose of X-ray required to reduce the cell survival to x% (Dx). The correlation coefficient of clones with 3-cycle treatment between CDDP and X-ray sensitivity increased gradually by increasing the end point of Dx from D10 to D90, resulting in significant correlation at D90. The result suggested that there is a certain common repair-related mechanism affecting both CDDP and X-ray resistance in CDDP-treated cells.

The mode of cell death (apoptosis or necrosis) in oral squamous cell carcinoma cell line (KM-1) exposed to Cisplatin was studied by various techniques including flow cytometry. All of the cells were killed by 48 hours' treatment with 100 microM CDDP. The proportion of the apoptotic cells was, approximately, only 95%, while the rest was necrosis. These results were demonstrated by observation of cellular morphology, agarose gel electrophoresis of DNA, flow cytometric analysis of propidium iodide (cell membrane integrity)/rhodamine 123 (mitochondrial activity), and flow cytometric measurement of sub-G1 peak population in DNA histogram. Although all the necrotic cells (91%) lost cell membrane integrity without DNA fragmentation, 46.9% cells revealed positive staining by TdT assay which identifies 3'-OH ends of DNA strands. These data suggest the possibility that the cytocidal mechanism of CDDP might involve single-strand breakage of DNA, but not double-strand breakage as occurred in apoptosis.

We undertook a pharmacokinetic comparison between cisplatin and nedaplatin in hepatic arterial chemotherapy using complete hepatic venous isolation and charcoal hemoperfusion (HVI.CHP). Dogs received a 20-min hepatic arterial infusion of either 4 mg/kg cisplatin (Group I, n = 5) or 4 mg/kg nedaplatin (Group II, n = 5) under HVI.CHP. The CHP filter adsorption ratios did not differ significantly between the two groups. Drug clearance fractions by HVI.CHP in group II (free pt 45.9%, total pt 55.1%) were significantly higher than those in group I (free pt 23.4%, total pt 27.5%) (p < 0.05). In contrast, hepatic extraction ratios in group I tended to be higher than those in group II. These results indicated that extrahepatic distribution of cisplatin became lower than that of nedaplatin, when given via the hepatic artery, and that the pharmacokinetic advantage of HVI.CHP would be greater when a drug like nedaplatin with a relatively low hepatic extraction ratio is used. Thus, we concluded that nedaplatin is a more suitable drug than cisplatin.

Chemoprevention of colorectal cancer has been extensively investigated in animal models and in high-risk human populations with inherited or acquired genetic changes, using anticarcinogenic agents from natural and synthetic sources. To understand active agents using a short-term assay, reliable intermediate biomarkers other than cancer are required as end-points. Endoscopically detectable aberrant crypt foci and adenomas are useful biomarkers in human intervention trials. Indomethacin and other nonsteroidal antiinflammatory drugs (NSAIDs) inhibit carcinogen-induced colon cancer development in rats. It was reported that a number of colorectal polyps in familial adenomatous polyposis patients regress after several months of sulindac treatment. Epidemiological studies have shown that regular use of aspirin and other NSAIDs reduces the risk of colorectal cancers and adenomas. In addition, ursodeoxycholic acid and alpha-difluoromethylornithine, a selective inhibitor of ornithine decarboxylase, have been employed in human intervention trials. Vegetable antioxidants such as carotenoids and flavonoids, omega-3 fatty acids, lactic acid bacteria, and indigestible oligosaccharides may also be promising chemopreventive agents.

We analyzed the pyrimidine metabolite in the urine of a patient with severe mucositis and hand and foot syndrome, who was administered 5-fluorouracil for recurrence of gastric cancer. From our analysis, it was suggested that the patient had decreased dihydropyrimidine dehydrogenase activity. Dihydropyrimidine dehydrogenase activity is usually measured in peripheral blood mononuclear cells, but this time it was estimated from the analysis of uracil, dihydrouracil, thymine, and dihydrothymine in the urine. We concluded that urinary analysis of the pyrimidine metabolism is effective as screening for the prediction and prevention of 5-fluorouracil toxicity.

We performed a clinical evaluation on the antiemetic profile and the plasma concentration of Azasetron Hydrochloride (a new selective 5-HT3 receptor antagonist), in transcatheter arterial chemoembolization using CDDP for unresectable hepatocellular carcinoma. Antiemetic effects were examined in 32 patients in the serotone group (administration of serotone 10 mg + methylprednisolone 125 mg) and in 77 patients of the control group (administration of metoclopramide 20-30 mg + methylprednisolone 500 mg). The response rate and the CR ratio in serotone group was 97% and 66%, respectively. These results were statistically higher than in the control group. Although all patients had chronic liver diseases, no side effects and complications related to administration of serotone were observed. The average area under the concentration (AUC) curve of plasma serotone in five patients with liver cirrhosis was 531 ng.h/ml, which was greater than that of a healthy volunteer. In conclusion, serotone is a new, safe and useful antiemetic drug in TACE therapy for hepatocellular carcinoma.