zinostatin and Colonic-Neoplasms

Mouse monoclonal antibody A 7, which was raised against human colon cancer, was used for preparing the conjugates with neocarzinostatin, mitomycin C and adriamycin. The tissue distribution of these three conjugates were examined in athymic nude mice transplanted with human colon cancer. The distribution in tumor was different in these three conjugates. The route of administration of the conjugate affected its distribution in tumor tissues. In the case of tumor transplanted in the back of mice, intravenous administration seemed to superior to intraperitoneal one. In clinical trials of immunoconjugate composed of A 7 and polypeptide anticancer drug neocarzinostatin (A 7-NCS), human anti-mouse antibody (HAMA) was observed in most cases without serious immune response such as anaphylactic shock. Human antibody against neocarzinostatin could not be detected in any case receiving A 7-NCS.

Topics: Animals; Antibodies, Monoclonal; Colonic Neoplasms; Doxorubicin; Humans; Immunotoxins; Mice; Mice, Nude; Mitomycin; Mitomycins; Neoplasm Transplantation; Tissue Distribution; Zinostatin

Polyanionid copolymer of divinyl ether and maleic anhydride (DIVEMA) with narrow molecular weight distribution was synthesized and tested of its antitumor activity. DIVEMA showed a significant antitumor activity against colon 26 adenocarcinoma and FSaI fibrosarcoma transplanted in syngenic mice. Furthermore, DIVEMA was used as a polymeric drug carrier of antitumor drugs to reduce side effects and enhance the antitumor activity of the drugs. Adriamycin and neocarzinostatin were attached covalently to DIVEMA and the polymeric conjugates showed higher antitumor activity than the corresponding mother drugs against P 388 leukemic mice.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Chemical Phenomena; Chemistry; Colonic Neoplasms; Doxorubicin; Drug Carriers; Humans; Leukemia P388; Macromolecular Substances; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Polyelectrolytes; Polymers; Pyran Copolymer; Zinostatin

The significance and effectiveness of adjuvant intraoperative chemotherapy using subserosal or submucosal administration of Neocarzinostatin (NCS) and CH 40 for gastric cancer and colorectal cancer were studied. Tissue NCS concentration of proximal lymph nodes were higher than for distant lymph nodes, while the immunocompetency of distant lymph nodes (lower NCS concentration) showed slightly higher activity than that of proximal lymph nodes. From these results, it is suggested that loco-regional administration of NCS might be effective for chemical lymph node cleaning of the cancer.

Topics: Carbon; Chemotherapy, Adjuvant; Colonic Neoplasms; Gastric Mucosa; Humans; Injections, Intralesional; Lymph Nodes; Lymphocyte Activation; Rectal Neoplasms; Stomach Neoplasms; Zinostatin

Cdc25C and p53 have been reported to be physiological targets of checkpoint kinase 2 (Chk2). Surprisingly, although Chk2 purified from DNA damage sustaining cells has dramatically increased ability to phosphorylate Cdc25C when compared with untreated cells, its ability to phosphorylate p53 is weak before treatment, and there is no increase in its activity toward p53 after DNA damage by gamma irradiation or the radiomimetic agent neocarzinostatin. Furthermore, introduction of Chk2 short interfering RNA into three different human tumor cell lines leads to marked reduction of Chk2 protein, but p53 is still stabilized and active after DNA damage. The results with Chk1 short interfering RNA indicate as well that Chk1 does not play a role in human p53 stabilization after DNA damage. Thus, Chk1 and Chk2 are unlikely to be regulators of p53 in at least some human tumor cells. We discuss our results in the context of previous findings demonstrating a requirement for Chk2 in p53 stabilization and activity.

Topics: cdc25 Phosphatases; Cell Cycle Proteins; Checkpoint Kinase 1; Checkpoint Kinase 2; Colonic Neoplasms; DNA Damage; Gamma Rays; Humans; Nucleic Acid Synthesis Inhibitors; Phosphorylation; Protein Kinases; Protein Serine-Threonine Kinases; Substrate Specificity; Tumor Cells, Cultured; Tumor Suppressor Protein p53; Zinostatin

To clarify the effect of SMANCS on malignant pleural carcinomatosis, seven patients with malignant pleural effusion were treated with SMANCS administered via an intracavitary route. Five patients showed improvement after one or two injections of SMANCS into the thoracic cavity, although 2 patients needed further therapy with the immunopotentiating agent picibanil (OK-432). No serious adverse effects were observed. This simple therapeutic tactic with SMANCS may be effective in cases of malignant pleural carcinomatosis.

Topics: Aged; Antineoplastic Agents; Breast Neoplasms; Colonic Neoplasms; Female; Humans; Lung Neoplasms; Male; Maleic Anhydrides; Middle Aged; Pleural Effusion, Malignant; Polystyrenes; Stomach Neoplasms; Zinostatin

Zinostatin stimalamer (ZSS) is a new anticancer agent derived from neocarzinostatin (NCS), which is synthesized by conjugation of one molecule of NCS and two molecules of poly(styrene-co-maleic acid). ZSS exhibited potent in vitro and in vivo antitumor activity in preclinical experiments, and a clinical trial of the intra-arterial administration of ZSS with iodized oil on hepatocellular carcinoma showed potent antitumor activity. We investigated the effect of ZSS and NCS on antitumor resistance and found that pretreatment with either drug suppressed the growth of MethA tumors in Balb/c mice and induced tumor eradication when given separately by single administration at therapeutic doses between 1 day and 4 weeks before tumor transplantation. The findings that the cytocidal activity of these drugs was not detected in vivo at the time of tumor transplantation and that tumor regression was preceded by a period of transient growth suggested that tumor regression was due to host-mediated antitumor activity induced by these drugs. Pretreatment with ZSS or NCS also suppressed the growth of Colon 26 carcinoma and Sarcoma 180. The finding that NCS showed the same effect as ZSS suggests that poly(styrene-comaleic acid) is not essential for the induction of host-mediated antitumor activity. Furthermore, apo-ZSS, which lacks cytocidal activity, did not induce antitumor activity. From this, it is suggested that the cytocidal effect of ZSS involves the induction of host-mediated antitumor resistance. In athymic Balb/c nu/nu mice, pretreatment with ZSS or NCS did not induce tumor eradication, suggesting that mature T lymphocytes play an important role in tumor eradication. Challenging MethA was rejected without transient growth in mice that had been cured of MethA, but challenging Colon 26 was not, showing that anti-MethA resistance was augmented selectively in the MethA-eradicated mice. Splenocytes from MethA-bearing mice pretreated with the drug showed tumor-neutralizing activity beginning 14 days after tumor transplantation. Tumor-neutralizing activity was only induced after MethA transplantation. The effector cells of this tumor-neutralizing activity were Thy1.2+ T lymphocytes that had been passed through a nylon-wool column, but no significant augmentation of cell-mediated cytotoxic activity of splenocytes from MethA-eradicated mice was observed in vitro.

Topics: Animals; Carcinoma; Colonic Neoplasms; Cytotoxicity, Immunologic; Drug Screening Assays, Antitumor; Female; Graft Survival; Male; Maleic Anhydrides; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred DBA; Mice, Inbred ICR; Mice, Nude; Neoplasm Transplantation; Polystyrenes; Sarcoma 180; Sarcoma, Experimental; T-Lymphocytes, Cytotoxic; Zinostatin

Topics: Animals; Antibodies, Monoclonal; Chimera; Colonic Neoplasms; Humans; Immunoglobulin Fab Fragments; Immunotoxins; Mice; Mice, Nude; Zinostatin

The murine monoclonal antibody (Mab) A7 conjugated to neocarzinostatin (A7-NCS) was injected intratumorally (IT) into tumor bearing nude mice. Its pharmacokinetics and tumoricidal effects were compared in the high, moderate and low antigen expressing xenograft for SW1116, WiDr and KB tumor-bearing nude mice, respectively. When injected IT into nude mice, [125I]A7-NCS was retained in the tumors according to the degree of antigen expression; it was also disseminated into the blood inverse proportion to the antigen expression. Addition of an excess amount of Mab A7 reduced [125I]-A7-NCS accumulation in SW1116 xenograft and elevated the [125I]A7-NCS concentration in the circulation. Complete tumor reduction was found in all 5 mice with SW1116 tumor, and 2 of 5 mice with WiDr tumor. However, only incomplete tumor suppression was observed in mice with the KB tumor. The significant tumor reduction in SW1116 bearing nude mice was attenuated when excess of Mab A7 was simultaneously administered with A7-NCS. These findings indicate that A7-NCS was localized in the target tumors and exerted its tumoricidal effects depending on the degree of antigen-antibody interaction when administered IT. Thus, A7-NCS can be used successfully in vivo for local therapy, auguring new and promising applications for local cancer therapy.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Antigens, Neoplasm; Carcinoma, Squamous Cell; Colonic Neoplasms; Humans; Immunotoxins; Injections, Intralesional; Iodine Radioisotopes; KB Cells; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Radioimmunoassay; Transplantation, Heterologous; Tumor Cells, Cultured; Zinostatin

Zinostatin stimalamer (YM881) is an antitumor agent, chemically synthesized by coupling one molecule of neocarzinostatin (NCS), with 2 molecules of styrene maleic acid half-butylester copolymer. YM881 showed strong cytotoxicity to human (KB, ST4 and others) and mouse (P388, L1210) tumor cell lines and also drug-resistant tumor cell lines. The antitumor effects were observed in murine MM46, colon 26 and other tumor models. The antitumor activity was as effective as NCS or better than NCS at the effective dose ranges.

Topics: Animals; Antineoplastic Agents; Colonic Neoplasms; Drug Screening Assays, Antitumor; Fibrosarcoma; Humans; Leukemia L1210; Leukemia P388; Maleic Anhydrides; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred DBA; Polystyrenes; Stomach Neoplasms; Zinostatin

The pharmacokinetics of a disulfide linked conjugate of a murine monoclonal antibody A7 with neocarzinostatin (A7-NCS) was studied following its intravenous administration to nude mice. Disappearance of the conjugate from the circulation was biphasic: an early rapid phase was followed by a much slower phase. The conjugate was removed from the blood circulation with a half-life of 12 hr, showing nearly the same kinetics as the free antibody. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the disulfide linkage in A7-NCS was stable at least for 48 hr after administration of the conjugate to nude mice. The conjugate concentration in a human colon cancer SW1116 derived tumor reached maximum at 24 hr after injection and remained high for an additional 24 hr. The passive hemagglutination inhibition assay revealed that NCS in the conjugated form can be efficiently delivered to the target tissue. The present report indicates that A7-NCS was sufficiently stable in circulation to reach the target tumor without releasing NCS.

Topics: Animals; Antibodies, Monoclonal; Colonic Neoplasms; Drug Carriers; Drug Stability; Half-Life; Humans; Injections, Intravenous; Iodine Radioisotopes; Mice; Mice, Nude; Neoplasm Transplantation; Neoplasms, Experimental; Radioimmunodetection; Transplantation, Heterologous; Zinostatin

We prepared an immunoconjugate, A7-NCS, of a mouse-derived anti-human colon cancer monoclonal antibody A7 and the macromolecular protein anticancer agent neocarzinostatin (NCS), and evaluated changes in human anti-mouse antibody (HAMA) by ELISA in the serum of patients intraarterially administered this agent. IgG and IgM class HAMA were detected in all patients, but no IgE class HAMA. In patients with stage V disease, the survival rate was higher in a group treated with A7-NCS at an NCS dose of 4,000 units or more than in that treated at an NCS dose of less than 4,000 units. In these patients, the survival rate was higher in a group treated with A7-NCS at an NCS dose of 4,000 units or more than in one not treated with it. These results suggest the usefulness of A7-NCS administration at high dose for prolonging survival of patients with advanced colon cancer.

Topics: Animals; Antibiotics, Antineoplastic; Antibodies, Monoclonal; Colonic Neoplasms; Humans; Immunoglobulin M; Immunotoxins; Mice; Survival Rate; Zinostatin

Monoclonal antibody-drug conjugates were applied as a clinical trial for patients who, based on the experimental study, had colorectal cancer. Monoclonal antibody A7, from a mouse splenocyte immunized against human colon cancer, was used as a drug carrier for colon cancer. The anti-cancer drugs mitomycin C (MMC) and neocarzinostatin (NCS) were bound covalently to A7 to form the conjugates A7-MMC and A7-NCS. The in vitro cytotoxic effects of the conjugates on SW1116 cells were stronger than those on free MMC or NCS. The conjugate A7-NCS, when administered to nude mice, brought about the highest NCS tumor concentration, whereas normal immunoglobulin G (IgG)-NCS distributed evenly in all tissues. The conjugates showed a strong antitumor effect on colon cancer transplanted into nude mice. Forty-one patients with colorectal cancer, including ten patients with postoperative metastasis, were given A7-NCS. The immunoperoxidase and drug concentration studies of the resected specimens showed that NCS was localized specifically in cancer. Patients receiving the conjugate did not experience serious adverse effects. Of the eight patients with postoperative liver metastasis, three showed evidence of tumor reduction on computed tomography (CT) scan and three claimed pain relief. The conjugate did not benefit patients with multiple lung metastasis or peritoneal metastasis.

Topics: Adult; Aged; Animals; Antibiotics, Antineoplastic; Antibodies, Monoclonal; Colonic Neoplasms; Drug Evaluation; Female; Humans; Liver Neoplasms; Male; Mice; Mice, Nude; Middle Aged; Mitomycin; Mitomycins; Neoplasm Transplantation; Rectal Neoplasms; Tumor Cells, Cultured; Zinostatin

Monoclonal antibody, A7, produced from a mouse splenocyte immunized against human colon cancer was used as drug carrier for colon cancer. A7 had not ADCC and ADMC activity but had ACD activity. Anticancer drug, mitomycin C (MMC), and neocarzinostatin (NCS), were covalently bound to A7 to form the conjugates, A7-MMC, and, A7-NCS. In vitro cytotoxic effect of the conjugates on SW1116 was much stronger than that of free MMC or free NCS. The conjugates, A7-NCS, administered in nude mice brought about the highest NCS concentration in tumor, while normal IgG-NCS distributed evenly in all the tissues. The conjugates showed strong antitumor effect on colon cancer transplanted in nude mice. Forty one patients with colorectal cancer including 10 patients with postoperative metastasis were given A7-NCS. The immunoperoxidase and drug concentration studies of the resected specimens revealed that NCS was found to be localized specifically in cancer. There was no serious adverse effect in the patients receiving the conjugate. Of eight patients with postoperative liver metastasis, three showed evidence of tumor reduction on CT scan and three claimed pain relief. The conjugate was of no benefit to the patients with multiple lung metastasis and peritoneal metastasis.

Topics: Adult; Aged; Animals; Antibiotics, Antineoplastic; Antibodies, Monoclonal; Colonic Neoplasms; Humans; Immunotoxins; Mice; Mice, Nude; Middle Aged; Mitomycin; Mitomycins; Neoplasm Transplantation; Rectal Neoplasms; Zinostatin

Neocarzinostatin (NCS) was conjugated with mouse monoclonal antibody (A7) that specifically reacted with human colorectal carcinomas in vivo and the conjugates (A7-NCS) were administered to patients with colorectal carcinoma. In several cases the size of the metastatic liver tumor was decreased after intraarterial infusion of A7-NCS. No severe adverse effects were observed in any patients who received A7-NCS. These results suggest that missile cancer chemotherapy using monoclonal antibody as a carrier of anticancer drug will be a promising method for use against cancer.

Topics: Antineoplastic Agents; Colonic Neoplasms; Humans; Immunotoxins; Infusions, Intra-Arterial; Zinostatin

In efforts to obtain preferential uptake of anticancer agents into tumors, we explored the possibility of delivering such agents by transferrin receptor-mediated endocytosis. Human diferric transferrin was conjugated with neocarzinostatin (NCS) using N-succinimidyl 3-(2-pyridyldithio)-propionate. This conjugate is capable of binding to the transferrin receptor and is internalized by endocytosis. The inhibitory effect of the conjugate on cell growth was remarkable when a human colorectal cancer cell line, M7609 was used as a target in vitro. In addition, in vivo efficacy of the conjugate in inhibiting the growth of M7609 cells implanted subcutaneously into nude mice was also observed when the conjugate was administered through a tail vein. The observed toxicity was a transient decrease in the red blood cell count, which returned to normal within 14 days. The half disappearance time of the conjugate was 55 min, while that of free NCS was 7 min. No serious side effects with regard to liver or kidney function were detected. This conjugate is an appropriate model for the receptor-mediated delivery of ligand-drug complex and may be useful for future clinical application.

Topics: Animals; Antibiotics, Antineoplastic; Colonic Neoplasms; Drug Carriers; Endocytosis; Humans; Mice; Mice, Nude; Neoplasm Transplantation; Receptors, Transferrin; Transferrin; Zinostatin

For targeting chemotherapy of colorectal carcinoma, mitomycin C (MMC) and neocarzinostatin (NCS) were covalently bound to monoclonal antibody A7 which is highly specific to human colon cancer. The in vitro cytotoxic effects of the conjugates A7-MMC and A7-NCS on SW1116 were 77 times and 4 times stronger than those of the free MMC and free NCS, respectively. An in vivo study in nude mice bearing human colon carcinoma revealed that monoclonal antibody A7 alone had no effect, and that A7-MMC and A7-NCS had greater inhibitory effects than the free MMC and NCS, respectively. Thirty-five patients with carcinoma of the colon and rectum including 6 with postoperative liver metastasis, one with postoperative lung metastasis and one with postoperative peritoneal metastasis, were given the A7-NCS conjugate consisting of between 15 and 90 mg of antibody and between 1,000 and 6,000 units of NCS. Immunoperoxidase study of resected specimens revealed selective localization of NCS in the cancer cells. The conjugate had no serious adverse effects. Five of the six patients with postoperative liver metastasis responded favorably to the conjugate, showing a decrease in tumor size on CT scan or relief of pain. The conjugate was of no benefit to patients with multiple lung metastasis or peritoneal metastasis. The effect on other patients with surgically resected carcinoma remains to be determined by a follow-up study.

Topics: Adenocarcinoma; Aged; Animals; Antibiotics, Antineoplastic; Antibodies, Monoclonal; Colonic Neoplasms; Female; Humans; Immunotoxins; Male; Mice; Mice, Nude; Middle Aged; Mitomycin; Mitomycins; Rectal Neoplasms; Zinostatin

Monocyte-derived macrophages (M phi) from cancer patients injected with low doses of Neocarzinostatin (NCS, 500 units/day, three times a week) 12 times produced significantly more superoxide (O2-) than controls. Lymphocyte functions, such as PHA response, surface marker and serum IAP, before and after NCS injections were the same. M phi from normal persons cultured with NCS (0.4 microgram/ml) for three days produced more O2- than controls, but those cultured with rINF gamma did not. These results suggest that the increased O2- production of M phi from patients taking low doses of NCS may be due to the direct action of NCS on the M phi.

Topics: Antibiotics, Antineoplastic; Colonic Neoplasms; Humans; In Vitro Techniques; Interferon Type I; Lymphocyte Activation; Lymphocytes; Macrophages; Monocytes; Neoplasm Proteins; Stomach Neoplasms; Superoxides; Zinostatin

Topics: Animals; Antibiotics, Antineoplastic; Catheterization; Colonic Neoplasms; Dogs; Fluorouracil; Humans; Iontophoresis; Rats; Rectal Neoplasms; Zinostatin