A late phase II study with MST-16 for malignant lymphoma was performed with the collaboration of 11 institutions belonging to the Hanshin Hematological Disease Treatment Research Group. Out of a total 40 patients entered into the study, 37 cases (33 of NHL and 4 HD) were evaluated for efficacy and 27 cases for side effects. The response rate was 29.7% (2 cases of CR and 9 PR). The factors affecting response rate were histological classification, phenotype of tumor cell surface and stage of disease. Bone marrow suppression and G-I disorders were the main toxicities observed, but they disappeared by the cessation of MST-16 treatment.

Nausea and vomiting induced by anticancer agents are common problems associated with chemotherapy for cancer. Recent trials have examined a variety of antiemetics, representing several different classes of drugs. High dose metoclopramide provided the impetus for many of the current studies because of its effect against cisplatin induced vomiting. However, current regimens are not yet entirely effective in many patients receiving cisplatin or other highly emetogenic anticancer agents. A promising new class of antiemetics, 5-hydroxytryptamine receptor antagonists are undergoing clinical evaluations. Members of this class are easily and safely administered to patients receiving cisplatin or other emetogenic anticancer agents. These are highly active antiemetics, both prophylactic and interventional treatment. Lack of extrapyramidal reaction and other adverse effects associated with its use makes the drug a very attractive one. However, studies of antiemetics require consideration of methodologic issues that may not be of concern in trials of anticancer agents. The results of these studies can be affected by the patient population, the sample size, pharmacologic variables, the trial design, the method of analysis, etc. Recently, developments both in new-antiemetics and better ways of using the existing ones, lead us to cautious thought that nausea and vomiting due to cancer chemotherapy can be controlled substantially with benefit to the patients.

The chemosensitivity test for esophageal and gastric cancer cells collected by endoscopic biopsies before operation was investigated for evaluation by ATP assay. Experimentally, ATP assay was applied in human esophageal and gastric cancer cell line transplanted in nude mice. ATP level was measured by Lumiphotometer and showed positive linear correlation with the number of cancer cells in more than 10(3). Also ATP level increased when more than 10(3) cancer cells were cultured for more than 48 hours. On the other hand, more than 10(3) cancer cells were indicated to be collected by endoscopic biopsies, experimentally. Clinically, 7 specimens collected by endoscopic biopsy and 5 anticancer agents (MMC, CDDP, 5-FU, ADM and BLM) were used for the test. Forty-nine cases, 31 cases of esophageal cancer and 18 cases of gastric cancer were subjected to the study. The evaluability rates were 93.8%, respectively. Over-all predictive accuracy for esophageal cancer between the clinical responses and results of the assay was 72.0%. These results suggested the usefulness of biopsy specimens for the chemosensitivity test of anticancer agents.

Recently, the semiautomated tetrazolium-based MTT colorimetric assay have been used to measure chemosensitivity. We also have been used this assay for 4 ovarian clear cell carcinoma cell lines to investigate the chemosensitivity of this tumor. In this study, several problems have been faced to be solved. In this paper, we pointed out these problems and indicated solutions.

In reviewing my lifework, I wish to summarize the results of studies on the development of new biological-active compounds. The following subjects are discussed: 1) syntheses and biological activities of benzoxazole derivatives, 2) structure-activity relationships of phyllodulcin, a natural sweetner, and 3,4-dihydroisocoumarins for sweetness, 3) syntheses and structure-activity relationships of spiro[isocoumarin-piperidine] analogs for antiallergic activity, 4) design, structure-activity relationships, and a mechanism of antitumor action of bistropolone derivatives, 5) design, structure-activity relationships of fused tetracyclic quinolines for antitumor activity.

It may not show accurate results if subrenal capsule assay (SRCA) is made only by measuring tumor size, because of infiltration of host inflammation cells resulted from host immune reaction. We developed a new method which make possible an accurate determination of chemosensitivity by measuring specific activity of succinate dehydrogenase (SD) of the tumor cells implanted in the subrenal capsular space. With reference to SDI test, the assay condition for measuring specific activity of SD was determined. A comparative study was carried out in which malignant tumors of the oral cavity serially transplanted in nude mice were tested with SRCA and subcutaneous transplantation assay in nude mice. Chemosensitivity to peplomycin (PEP), CDDP and 5-fluorouracil (5-FU) evaluated SSDI method and nude mouse assay showed a high correlation than those evaluated by TGIR method and nude mouse assay. The overall predictive accuracy compared with nude mouse assay was 72.2% by TGIR method and 88.9% by SSDI method. SSDI method seemed to be a useful method to evaluate the chemosensitivity in SRCA.

A cell line was established from a patient with malignant fibrous histiocytoma, which had originated in the maxillary sinus. Using this cell line, sensitivity to chemotherapeutic agents and cytotoxicity against various immunoeffecter cells were tested.

We describe our experience with succinic dehydrogenase inhibition (SDI) test for solid tumors as a chemosensitivity test using 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide. Specimens were obtained from 76 surgical resected tumors, including 32 colon cancers, 24 stomach cancers and 16 lung cancers. Following enzymatic dissociation of scissors-minced tumors, viable cells were cultured in serum free medium (S-Clone SF-B) for 4 days with eight drug concentrations obtained by 2-fold dilution of drugs. Among 76 specimens tested, 48 specimens including 15 colon cancers, 18 stomach cancers and 15 lung cancers were successfully evaluated. For the purpose of judgement, 50% inhibitory concentration (IC50) was calculated in each case. Tumor specimen was regarded as sensitive to a given agent when the IC50 value was the same or smaller than the cut-off concentrations (1 microgram/ml for mitomycin C, 5 micrograms/ml for cisplatin, 2 micrograms/ml for adriamycin and 50 micrograms/ml for 5-fluorouracil), and was regarded as resistant when it was larger than these levels. In vitro vs in vivo drug sensitivity was successfully evaluated in 23 cases. The overall predicting accuracy rate was 78% (18/23), with one true positive, 5 false positive and 17 true negative cases. This test appeared to be useful to tailor effective agents for patients because of its relatively high successful and predictive rates.

Five anticancer agents (MTX, 5FU, MMC, ADR and VCR) and an antiestrogenic agent (TAM) were investigated to observe the effects on the growth of MCF-7 cells and the changes on the cell cycle distribution of them by flow cytometry. The growth-inhibitory effects of these agents were divided into three manners. The growth rate by ADR was affected in a concentration-dependent manner; MTX in a time-dependent; 5FU, MMC and VCR in both time and concentration-dependent manners. The effects on cell kinetics were divided into three categories. MCF-7 cells were accumulated on S phase by MTX, G0G1 phases by TAM and G2M phases by the other agents. The combined effect of two agents was the most prominent when MTX was first administered and followed by an agent which affected on G2M phases. In the combination of TAM and MMC, the strongest effect was obtained when MMC was first administered and followed by TAM. These results indicated that the combined effect of cell inhibition was obtained on cell kinetic basis by employing the technique of analytical flow cytometry.

The chemosensitivity of 23 human head and neck squamous cell carcinomas (9 pharyngeal, 5 laryngeal, 5 oral cavity, 2 maxillary sinus, 1 salivary gland, 1 cervical esophagus) were examined using subrenal capsule (SRC) assays against 5-FU, FT, UFT and HCFU. Assays adequate for evaluation were obtained in 19 of 23 cases and the averages of tumor size changes on day 6 were, 17.3% for control, -0.8% for 5-FU, 10.3% for FT, -10.4% for UFT and 2.4% for HCFU. Sensitivity rates for each drug were 26.3% for 5-FU, 6.3% for FT, 38.9% for UFT and 11.1% for HCFU. To evaluate the clinical utility of these anticancer drugs, we determined the therapeutic index (TI), the ratio between tumor chemosensitivity and changes in animal body weight as a criterion for toxicity. The mean values of TI were 0.891 for control, 0.989 for 5-FU, 0.944 for FT, 1.054 for UFT and 0.980 for HCFU. UFT showed a greater antineoplastic effect as compared to the other three agents. Immunohistochemical study of the transplanted tumors revealed a specific antigen staining pattern similar to that observed in the original human cancer tissues. Degeneration of tumor tissues in drug-treated animals was also confirmed by immunohistochemical staining. Based on the results obtained from subrenal capsule assays, either by examining tumor size changes or evaluating TI, UFT proved to be the most effective of the 5-FU analogues against human head and neck squamous cell carcinomas.

In order to establish a sensitivity test system for the evaluation of anti-cancer hormonal agents, we tried a long-term subrenal capsule (SRC) assay, using nude mice with a transplanted solid tumor of endometrial cancer cells (Ishikawa's line). Unlike DNA-affecting agents, anti-cancer hormonal agents exert cytostatic effects rather than cytocidal effects, and their evaluation in a short period is considered to be inaccurate. Our test system is somewhat difficult in terms of technique, but it is useful since it can (1) evaluate the agents in the relatively short period of 28 days and (2) compare the cytostatic anti-tumor efficacy of two or more agents under the same conditions. The rate of successful tumor transplantation in nude mice in our system is very high, i.e., more than 90%. Although there are some points which need improvement, our system is considered to be useful as an assay system for the development of anti-cancer hormonal agents and other similar chemotherapeutic agents for cancers. When medroxyprogesterone acetate (MPA) was evaluated by means of this system, the administration period, as well as the dosage, was found to be important.

Ten patients with malignant diseases whose mean age was 20.0 +/- 13.2 years received anthracycline derivatives therapy and were evaluated for their left ventricular systolic and diastolic functions by computer-assisted digitized M-mode echocardiography. Fractional shortening (%FS), a parameter of systolic function, was measured. The first derivative of left ventricular dimension change (peak LV dD/dt), posterior wall thinning (peak LVPW thinning rate) and interventricular septum thinning (peak IVS thinning rate) were used as indices of diastolic function. Blood pressure (BP) was measured noninvasively at the end of the echocardiographic examination and hemoglobin concentration (Hb) was measured on the same day. These examinations were performed immediately before administration of anthracycline and one week and one month after the last administration. Statistical analyses were performed using the Student's t-test. The mean BP, HR, LVDd, LVDs, LVPW and IVS remained unchanged following the drug administration. %FS did not change significantly; 36.8 +/- 6.3%, before the administration, 35.3 +/- 6.5%, one week after the administration, and 36.5 +/- 5.1%, one month after the administration. Peak LVdD/dt and the peak LVPW thinning rate decreased appreciably from 4.46 +/- 1.10 to 3.76 +/- 1.08, and from 7.99 +/- 1.55 to 6.41 +/- 1.04, respectively, one week after the administration. The peak IVS thinning rate decreased from 3.54 +/- 0.81 to 2.99 +/- 0.79 after one week (p < 0.01). All of these values returned to the control levels in one month after the drug administration. We concluded that the indices of left ventricular diastolic function were more sensitive for detecting cardiac impairment than those of systolic function during the course of anthracycline therapy.

The usefulness of 5'-DFUR in both patients with uterine cervical and ovarian cancers was investigated by determining pyrimidine nucleoside phosphorylase (PyNPase) activities and 5-FU levels in cancerous and normal tissues resected from them after oral administration of 5'-DFUR. In uterine cervical cancer, each group of 9 cases administered single dose of 400 mg of 5'-DFUR and 7 cases administered 400 mg of 5'-DFUR 3 times a day continuously for 7 days was investigated. In ovarian cancer, all of 9 cases were administered 400 mg of 5'-DFUR 3 times a day continuously for 7 days. In conclusion, PyNPase activities in the tissues of uterine cervical and ovarian cancers were higher than those in the normal tissues. 5-FU tissue levels in the cancerous tissues were significantly higher than in the normal tissues and blood as well. This tendency was observed in each of the single and continuous administration groups. These results suggest that the tumor selectivity which is one of characteristics of 5'-DFUR could be expected also for cancer in the field of gynecology.

The purpose of this study is to assess effects of fibroblasts in the vitro chemosensitivity testing on human lung cancer cells and to remove them. Fourteen lung cancer cell lines and 14 fibroblasts derived from resected specimens of lung cancers were used, whose S.D (succinate dehydrogenase) activities were measured with MTT colorimetric assay. The chemosensitivity of a lung cancer cell alone was compared with that of mixed cancer cell and fibroblast. As results, S.D activities of fibroblasts were less 2-4 fold than those of lung cancer cells. Fibroblasts were as sensitive to CDDP, MMC and 5-FU as lung cancer cells, but more sensitive to ADM and VP-16 than them. When sensitivity testings were performed on mixed cancer cells and fibroblasts, or mixed cancer cells and conditioned media of fibroblasts to CDDP with 3 day's incubation times, the sensitivity was affected in 61%, or 10% of all the pairs, respectively. However, when these tests were done without any incubation times, the sensitivity was not affected. Therefore, it was suggested that anticancer drugs had to be simultaneously added when single cell suspensions were plated if resected specimens were used in a anticancer drug sensitivity test.

The pharmacokinetic properties of KRN 8602, an anthracycline compound, was studied by HPLC following intravenous administration of KRN 8602 to cancer patients. The results were as follows. (1) The plasma concentration-time curve declined as a triphasic function (alpha, beta, gamma) (t1/2 (alpha) = 0.02910, +/- 0.0054 hr, t1/2 (beta) = 0.704 +/- 0.319 hr, t1/2 (gamma) 8.37 +/- 1.37 hr). The blood cell concentration was higher than that in plasma. (2) The distribution volumes of the tissue compartment were larger than those of the central compartment. This result suggested that KRN 8602 would be easily transferred into the tissues. (3) The area under the curve (AUC) of KRN 8602 increased in proportion to the increase of dosage. (4) The metabolites of KRN 8602 were detected in plasma, blood cell and urine. (5) Urinary excretion of KRN 8602 and its metabolites were extremely low.