In this study, we treated severe infections (21 cases) accompanied with induction chemotherapy in 20 patients with acute leukemia by the combination of a large dose (600 approximately 1,200 mg/day) of amikacin with other antibiotics. Infections during induction chemotherapy of acute leukemia consisted of sepsis (8 cases), pneumonia (7) and others (6), and most of causative organisms were Gram-negative bacteria, such as Ps. aeruginosa (7 strains), Flavobacterium (5), Serratia (3), Ps. cepacia (2), E. coli (2) and others. The combination chemotherapy of a large dose of amikacin with other antibiotics was found to be effective (71.4%) for such infections. Side effects were negligible except for drug eruption. Therefore, a large dose of amikacin should be given for the treatment of severe infections accompanied with induction chemotherapy of acute leukemia.

Phase I study of a new antitumor antibiotic, neothramycin which belongs to anthramycins was performed by a cooperative study group involving 15 major institutions. A total of 63 cases including 42 various solid tumors and 21 hematologic tumors refractory to standard treatments underwent the study during June 1979 to June 1981. Doses of single injection were escalated from an initial dose of 2 mg/m2 estimated by one twentieth of LD10 in mice up to 60 mg/m2. The most frequent and severest toxicity was nausea and vomiting seen in about the half of patients being administered dosages ranged from 24 mg/m2 to 40 mg/m2, and 3 out of 4 patients received dosages exceeding 50 mg/m2 required clinical managements; moreover, 1 out of 2 patients administered 60 mg/m2 was ranked as grade 4 of the criterion of toxicities in WHO handbook. Other clinical toxicities such as skin rash, hepatotoxicity or nephrotoxicity observed in the minority of the patients were reversible. Furthermore, hematologic toxicity was extremely mild and appeared not to be dose dependent. One patient with chronic myelogeneous leukemia had a hematological improvement and the other with esophageal cancer had a partial response. The result indicates that a dose limiting factor of neothramycin is nausea and vomiting, and a maximum tolerated dose of a single injection is 60 mg/m2. A dose schedule of 30-40 mg/m2 appears to be an optimal dose for Phase II study.

A phase II study of a new fluoropyrimidine antitumor agent, carmofur (HCFU) fine granules, was performed in 24 institutions. The eligibility of patients and the criteria of response evaluation were based on the "Japanese Criteria for Evaluation of Clinical Effects of Cancer Chemotherapy on Solid Tumors." Out of 119 patients entered in the study, 65 patients were evaluable: 63 patients among them had measurable or evaluable lesions. Positive responses better than PR (Partial Response) were obtained in five (17.9%) out of 28 patients with gastric cancer, four (36.4%) out of 11 patients with colorectal cancer and five (45.5%) out of 11 patients with breast cancer. The total positive response rate was 22.2%. Optimal doses for the clinical use were ranged from 9 to 18mg/kg, and the median duration of PR was 19.6 weeks. In addition to the similar gastrointestinal toxicities observed in other fluoropyrimidines, hot sensation, pollakiuria and frequent defecation were seen in 12.4%, 10.1% and 3.4% of patients, respectively.

Human motor endplate acetylcholinesterase was inhibited in vitro by alkylating antineoplastic agents, most strongly by mechlorethamine, followed by DTIC, ACNU, cyclophosphamide and ifosfamide. Eleven other antineoplastic agents did not inhibit the enzyme substantially nor interfered with cholinesterase measurement. Cyclophosphamide and mechlorethamine inhibited human plasma pseudocholinesterase most strongly, followed by thiotepa, ACNU, DTIC, ifosfamide and BCNU. Mechlorethamine, ACNU and ifosfamide inhibited the motor endplate and plasma cholinesterase practically equally, DTIC inhibited motor endplate cholinesterase more strongly, while cyclophosphamide was a more selective inhibitor of plasma cholinesterase. Inhibition of human red blood cell acetylcholinesterase was identical to that of motor endplate acetylcholinesterase; therefore, red cells would be a preferable indicator in monitoring cholinesterase inhibition by antineoplastic agents.

The effects of various anticancer agents on human choriocarcinoma transplanted to nude mice, specifically, inhibition effects on tumor growth and survival rate were studied to establish an appropriate chemotherapy for refractory choriocarcinoma. The agents studied were methotrexate (MTX), actinomycin D (ACD), cyclophosphamide (CPM), vincristine (VCR), L-PAM (MPI), bleomycin (BLM), carboquone (CQ), cisplatin (CDDP), ACNU, MCNU, vinblastine (VLB), VP16-213, OK-432, Maruyama vaccine (SSM) and metronidazole (ME), and the following results were obtained: 1) The inhibition effects on tumor growth were obtained in the groups of VCR, VP16-213, CDDP, MPL, CPM and CQ; 2) The survival rate was 100% in the groups of MPL, BLM, and CQ. In the groups of ACD VLB, VP16-213 and VCR, all mice died. 3) MPL was found to be the most effective agent in terms of inhibitory effect and survival rate. In the future, combination chemotherapy including MPL and maintenance chemotherapy with MPL to refractory choriocarcinoma should be studied.

The usefulness of the nude mouse-human cancer xenograft system for a sensitivity test of chemotherapeutic drugs is evaluated. In our present study, the response of 9 experimental chemotherapies on 7 lines of cancer xenografts in nude mice was directly compared with clinical response in each donor patient, to the same chemotherapy and good correlation was found between these respective results. Experimental studies on chemotherapeutic effects on 14 lines of cancer xenografts (7 gastric, 3 colorectal, 3 breast and 1 pancreatic cancer) using 5 clinically active and 5 new drugs, were also conducted Although every line of xenograft retained different patterns of sensitivity to the various agents, drugs which were clinically effective to a type of tumor were found to be also sensitive against the same type of xenografts in nude mice. Thus the nude-mice human cancer system were is thought to be useful as a predictive secondary screening for new drugs. However, further improvement is necessary to establish this system as a more accurate selection method for anti-cancer drugs.

Human tumor stem cell assay is an in vitro colony-forming technique. Double soft agar layers are used for culture tumor cells and cell lethality is judged by the numbers of colony formation in this assay. Single-cell suspension made from various malignant materials in cancer patients is placed in culture after exposing to various anticancer agents for one hour and incubated for two weeks. Antitumor effects of various anticancer agents against individual patients are evaluated by % inhibition of colony formation. Of 57 tumor specimens 42 (74%) formed at least five colonies per plate (per 0.5 x 10(6) cells). The colony-forming rates of various malignancies are as follows: breast cancer 14/15 (93%), ovarian cancer 8/10 (80%), stomach cancer 5/13 (38%), sarcoma 4/5 (80%), lung cancer 1/4 (25%), colon cancer 3/3, each of pancreas cancer, leukemia and primary unknown adenocarcinoma 2/2, malignant lymphoma 1/1. The median plating efficiency (number of colonies/number of nucleated cells plated) is 0.02% (range: 0.001-0.3%). High correlation between human tumor stem cell assay results and response of an individual patient's tumor to chemotherapy is reported by Salmon and Von Hoff. Human tumor stem cell assay is useful tool for the high prediction of chemosensitivity response.

The sensitivity of anti-cancer drugs against cultured human lung cancer cells and the first cultured xenograft tumor has been measured by the microcolonies inhibition test in microplates (Falcon, Micro Test II). The results obtained were as follows; 1). The drug sensitivity of cancer cells has differed in each case. The established cancer cell lines of the same cell type and the same growth speed have also showed a different sensitivity. Therefore, it suggests that the measurement of drug sensitivity of cultured human cancer cells is usefull for the clinical application. 2). The drug sensitivity of cancer cells has been elevated when they changed their character during the passage cultures. On the other hand, the successful culture of cancer cells has been elevated when they were purified with discontinuous density gradients and a hypotonization. 3). In future, further efforts to develop a better medium for the colonies inhibition test, i. e., a conditioned medium containing the specific growth factor for the colony of cancer cells are imperative.

A sensitivity test of antitumor drugs using primary shortterm culture with 3H-thymidine was performed on stomach cancer patients. Stomach cancer tissues, which were removed in the operating room, were cut into cubes of 1 to 2 mm. Thereafter, they were put on a stainless steel mesh which was placed in a small petri dish (40 X 15 mm) and they were moisted with the cultivating medium. An appropriate dose of antitumor drugs (MM-C, 5-FU, cytosine arabinoside) was added at the onset of cultivation and 3H-thymidine was further added 24 hours later. At the end of cultivation in a CO2 incubator for about 3 days, the specimens were homogenized in an ice-cold homogenizer with 5% trichloroacetic acid. DNA fraction of the specimens was extracted by Schmidt-Thannhauser method. The isotope activity in DNA fraction was measured in a scintillation counter and was represented as cpm/microgram DNA. Sensitivities of antitumor drugs can be determined by comparison between isotope incorporations into the specimens tested and those into control specimens. Bacterial and fungal contaminations were observed in primary stomach cancer tissues.

A test system using dehydrogenase activity for predicting the response to chemotherapeutic agents against cancer cells was introduced. Agar plate assay, INK which test, and SDI test commonly employed in the clinical study were also reviewed. Agar plate assay resembles antibiotic disc sensitivity test. The cancer uniformly suspended in the agar medium was exposed to drugs on paper discs for few hours. After removal of the disc, methylene blue or 2, 6-dichlorophenol indophenol was applied as a dehydrogenase indicator. INK test was introduced by Nishioka et al. in 1957. Several fragments of fresh cancer tissue were incubated with chemotherapeutic agents in roller test tubes. Twenty-four hours later, 2, 6-dichlorophenol indophenol was applied as a dehydrogenase indicator. To develop a simple, rapid, and comparable test, SDI test was introduced by us in 1964. A fresh cancer tissue was minced and made into the cell suspension. After cancer cells were exposed to chemotherapeutic agents, the activities of succinic dehydrogenase of the treated cells were measured by the reduction of 2, 3, 5-triphenyl tetrazolium chloride. Some points to be improved were investigated and discussed.

Many anticancer drugs are now available for clinical use. Unfortunately, most are extremely toxic to normal tissue. Use of one or more toxic drugs inactive against the given patient's own cancer may not only deny the patients the benefit of active therapy but may actually make the patient's cancer grow faster. The need for adjuvant chemotherapy can scarcely be doubted. However, distant metastases may be present at the time of primary surgery in many cases. Some form of generalized therapy like drug therapy or immunotherapy must therefore be used to supplement our local form of treatment such as surgery and radiation therapy for cancer. Already the dangers of cancer chemotherapy are reported as adverse effect, not only in the case of advanced cancer patients, but, that of post operative adjuvant chemotherapy. Therefore, predictive assays analogous to antibiotic sensitivity tests are needed to rule out inactive drugs and select active drugs with least toxicity. The treatment of patients with cancer according to the result of tests of sensitivity of cancer cells to anticancer drugs has been introduced many years ago. The examination of the sensitivity is carried out on explanted tumor cells by two different methodical approaches-observation of the cytotoxic effect of the preparation tested to the growth of the tissue culture of the tumor examined, and a short-term examination with the indication of the effect of anticancer drugs according to the decreased utilization or incorporation of the precussors of the synthesis of proteins, RNA and DNA labeled with radioisotopes, or according to the release of enzymes from tumor cells damaged by an efficient anticancer drug. However, clinicians know that there are many disagreements between the sensitivity test and clinical results. Each sensitivity test has its limitation and problems for resolution. Research toward the goal of the sensitivity test is to divise more appropriate method which is simple prompt and precise for drug selection.

An attempt has been made to construct an assay of anti-cancer drug sensitivity which is suitable for use with primary culture cells from human bone and soft tissue tumors. The ability of cells to incorporate labeled nucleic acid precursors into acid precipitable material was assessed. Hela-S3 cells were employed to avoid inherited variability and heterogenity of primary cultures of human tumors. Labeled nucleic acid precursors were used not to assay the changes of DNA or RNA synthesis but to detect the viability of cells. A logarithm of counts per minute of incorporated labeled precursors is in proportion to a logarithm of viable cell numbers. This relationship was not influenced by the labeled precursor incubation time. Multiplate was used to provide large numbers of replicate cultures without the requirement of a large numbers of cells. Monolayer Hela-S3 cells which were seeded 24 hours earlier were incubated for three hours with various concentrations of drugs. After removal of drugs, cells were cultured for one week. In this period, Hela-S3 cells with no drug treatment became almost confluent and mitoses occurred about four times. Labeled precursors were incubated for three hours, and incorporated 3H-thymidine was counted. A standard curve of incorporated labeled precursor counts and viable cell numbers was drawn for every assay. The density inhibition of labeled nucleic acid precursor incorporation can be checked and connected with the standard curve, and viable cell numbers after drug exposure can be obtained from the standard curve. When percent survival is plotted against drug concentration, a sigmoid curve is obtained if the drug has dose dependent effect and then the 90% lethal dose (LD90) can be determined from the curve. LD90 was used as the index of anti-cancer drug sensitivity. If the drug has time dependent effect, percent survival of maximal inhibition was used as the index of drug sensitivity.

A new fluorinated pyrimidine, TAC-278, was studied for its safety, anti-tumor activity and pharmacokinetics in patients with solid tumors of various types. Single oral administration was done in 43 patients with dose range of 50 to 1200 mg. These single administrations caused no side effects but nausea in only one patient daily given 900 mg. Repeated oral dose tolerance was assessed in 79 patients in daily doses of 100 to 1800 mg. Side effects were reported by 26 (33.3%) of the 79 patients. Major side effects were mild gastrointestinal symptoms. The maximum tolerated dose was considered to be 1200 mg/day, over which CNS symptoms as dose limiting factor for TAC-278 developed in some patients. As to the therapeutic effect, minor response was obtained in 2 of the 24 evaluable patients. Concentration of 5-FU in body fluid and tumor tissues were high following oral administration of TAC-278, but the disappearance was relatively rapid. Excretion of TAC-278 occurred predominantly in urine.

Anthracycline antibiotics are principal agents in the treatment of acute non-lymphocytic leukemia, although the usefulness are limited by their adverse side effects, especially by the cardiotoxicity. Aclacinomycin A (ACM) is known to be a new anthracycline antibiotic which has been isolated from Streptomyces galilaeus, and its cardiotoxicity on the experimental animal systems was reported to be more than 10 times lower than that of adriamycin. We investigated the cardiotoxicity of ACM on 29 patients with acute leukemia and compared it with daunorubicin (DNR). The measurement of STI (PEP:LVET) has been recommended to be convenient method of assessing the anthracycline cardiotoxicity, but through out analytical study, QTC measurement was proved to be more valuable for the simple and rapid detection of the cardiotoxicity induced by the agents. In comparison with the QTCs in DNR and ACM, the cardiotoxicity of ACM was much lower than that of DNR, and the reversibility of ACM induced cardiotoxicity was much more rapid. Moreover, these effects were observed even in the patients treated with the maximum dose of DNR. Therefore, ACM was expected to be one of the agents of the first choice for the relapsed cases of acute leukemia, especially APL.

UFT (a mixture of futraful and uracil) was administered orally to 15 cases of recurrent and superficial bladder cancer principally for more than 4 weeks at the doses of 300 or 600 mg per day. The result was evaluated by Koyama-Saito's response criteria. Of 14 evaluable cases, complete remission was noted in 5. Two cases of advanced bladder cancer were treated with UFT combined with irradiation therapy, and one case was found that cancer cells degenerated severely in the primary lesion pathologically. Another case was recognized that lung metastasis disappeared completely. As the side effects, anorexia occurred in 2 cases with 600 mg per day and 1 case with 300 mg, but it was not so severe to terminate UFT therapy. From above-mentioned results, UFT seems to be a useful drug for the treatment of bladder cancers.

In 26 patients with cancer of the stomach, bone marrow function, cell mediated immunity and plasma CEA level were examined after administration of ACN-U. ACNU was given intravenously and intermittently with 5-FU given orally daily. Results 1) Delayed myelosuppression was observed in every cases and reached to the nadir in 2-7 weeks after ACNU administration. 2) Bone marrow suppression and recovery therefrom were observed earlier in platelet counts than in RBC and WBC counts. 3) Suppression of cell mediated immunity was not observed later than 4 weeks after ACNU administration. 4) Plasma CEA level decreased or stopped to increase after ACNU administration in 5 out of 8 patients having a high CEA level before treatment. 5) Regression of tumor size was observed in 4 out of 5 patients in whom a tumor was palpable. Conclusion ACNU has a strong antitumor activity and may be more effective if combined with antimetabolite agent, such as 5FU. Total dosage of ACNU given safely in one series was considered to be 150-200mg. Next series of ACNU administration should be started after restoration of platelet counts more than 100,000/mm3 was obtained.

Chemotherapy of advanced bladder cancer aims to destroy all the cancer cells in the host. For this purpose the most suitable and effective anticancer agents should be chosen. There have been many methods to select the anti-cancer drugs: sensitivity test. However, no reliable tests are available. We developed new anti-cancer sensitivity test, using the radio-active nucleic acids precursors; C14-Formate and C14-Adenine. This test revealed that Cisplatin, Adriamycin, and Mitomycin C were the most potent for the transitional cell carcinoma of the urinary bladder. Chemotherapy with a single agent was disappointing. Combined use of these agents was rather promising. Among them combination of cis-platin with Adriamycin and/or cyclophosphamide was the most effective. However, the overall response rate was reported around 50%. Multi-disciplinary treatment including surgery, irradiation, chemotherapy, and immunotherapy was disclosed to be useful for the treatment of bladder cancer. Since 1977 25 cases were treated with this mode of therapy in our clinic. Anti-tumor effect was remarkable. The categories, disappeared, and over 50% decrease of the mass, were found in 96% of the patients. Also, down-staging was demonstrated in 20% of the cases. Histologically no cancer cells were found in the surgical specimens of 3 cases and no viable cancer cells in 3 cases respectively. From these results it is now assumed that multi-disciplinary treatment is promising for the treatment of bladder cancer.

The effect of preoperative bladder instillation (POI) and periodic prophylactic bladder instillation (PPI) of anticancerous drugs was evaluated in connection with the prevention of recurrent bladder tumors after surgery. A total of 191 patients with pTa or pT1 tumors including patients submitted to TURbt and partial cystectomy from January, 1967, to December, 1980, was chosen for the study. They were divided into the following 4 groups: Group A (49 cases) was treated with PPI and POI, Group B (11 cases) with PPI but not POI, Group C (46 cases) with POI but not PPI and Group D (85 cases) with neither PPI nor POI. POI was performed three times a week for a total of 20 applications of anticancerous drugs from two months before surgery. PPI was performed twice a month from one month after TURbt or parital cystectomy with a combination of 20mg of Mitomycin C and 1,000mg of 5-FU. The non-recurrence rate in these 4 groups was estimated by the actuarial method. The 3 year non-recurrence rates in Groups A,B,C and D were 95.4%, 90.9%, 44.0%, and 45.6%, respectively. The 5 year non-recurrence rates in Groups A,B,C and D were 82.4%, 81.0%, 32.0% and 35.1%, respectively. It is presumed from our study that PPI was effective in preventing the recurrence of bladder tumors. In comparison, POI showed a very limited effect and only in the first two years after surgery. No carcinogenic action on the bladder epithelium was observed from the topical use of Mitomycin C.