Influence of OK-432 on intratumor 5-fluorouracil concentration in cases given UFT

The influence of OK-432 on the activation of UFT, consisting of tegafur and Uracil, was examined in patients with gastric cancer and colonic cancer. In 14 gastric cancer and 15 colonic cancer cases, to which UFT 400 mg/day and OK-432 2KE 2/W were administered orally and intra-muscularly for 2 weeks preoperatively until surgical treatment, intratumor 5-fluorouracil (5-FU) concentration was measured and compared with that of patients who given UFT 400 mg/day orally for 2 weeks (15 gastric cancer and 15 colonic cancer cases). As the results, in the groups of patients given OK-432, the concentration in gastric cancer tissue was 0.093 +/- 0.067 microgram/g and that in colonic cancer was 0.098 +/- 0.058 microgram/g. Both values exceeded 0.05 microgram/g which is considered to be the effective intratumor 5-FU concentration. No difference was observed in these cases given UFT alone. The ratio of intratumor 5-FU concentration vs. that of normal tissue was 2.5 for gastric cancer and 2.7 for colonic cancer and the ratio of tumor vs. serum 5-FU concentrations was 8.5 for gastric cancer and 10.9 for colonic cancer. No difference was also observed in these values in cases given UFT alone. From above results, it seemed that the clinical dose of OK-432 2KE 2/W had no influence on the activation of UFT, so that the combination therapy of UFT and OK-432 was found to be clinically useful.     

Synergistic activation of rat alveolar macrophages by cepharanthine and OK-432

We studied the synergistic activation of rat alveolar macrophages (AM) by cepharanthine (Ceph.) and OK-432. Rat AM could be rendered much more tumoricidal against Walker-256 tumors in vitro after the i.v. injection of ceph. (5 mg/Kg) and OK-432 (5 KE/rat) thao of ceph. or OK-432 only. Radioactivity of 14C-acetate-OK-432 trapped in murine lung after the i.v. injection of mixed of ceph. was much higher than of 14C-acetate-OK-432 only. Rat AM could be rendered tumoricidal by incubation in vitro with OK-432, but not with ceph. This finding suggests that if OK-432 is injected intravenously with mixed of with ceph., of OK-432 is trapped much more up by the lung without mixture, therefore AM can be much more tumoricidal. Intravenous administration of OK-432 with ceph. may be useful for reduction of lung metastasis by tumoricidal AM.     

A case of sigmoid carcinoma successfully treated with endoscopic topical infusion of OK-432 and UFT oral treatment

A 78-year-old man had a tumor of Borrmann type II affecting about one third the circumference of sigmoid colon, which was diagnosed as well-differentiated adenocarcinoma from biopsy. Although surgical intervention was recommended and the radical operation seemed possible from preoperative examination, the patient first refused it. Of necessity, systemic administration (s.c) and single endoscopic topical infusion of OK-432 and oral treatment of UFT were performed for 1 month. Since the patient agreed to receive an operation later, sigmoidectomy and lymph node dissection (R 2) were performed, resulting in the disappearance of tumor cells histologically. One week before endoscopic infusion, oral UFT and subcutaneous OK-432 were initiated. Although no macroscopic change was found during the topical infusion as compared with the time of first examination, the bulging disappeared, showing a polypoid change and IIc-like findings after 3 weeks. The operation was performed after an additional week, after which a benign tumor was found macroscopically and the disappearance of tumor cells histologically. Although many reports have dealt with the effect of endoscopic treatment for gastric carcinoma, there have been fewer reports describing the endoscopic treatment performed in patients with colon carcinoma, making it difficult to evaluate its therapeutic effect. However, this endoscopic therapy was suggested to be capable of becoming a useful treatment for inoperable colon carcinoma from the present case, whereas chemotherapy was found to have almost no effect on colon carcinoma in general.     

Antitumor effect of OK-432 (3)--mechanisms of tumor growth inhibition by OK-432 induced activated macrophages

Spleen cells and peritoneal exudate cells obtained from BALB/c mice which had received an i.p. injection of 0.1 mg of OK-432 4 days previous to sacrifice, were examined by Winn's neutralization assay for their antitumor activity against Meth-A sarcoma cells in BALB/c mice. Both of the cell preparations clearly inhibited the growth of admixed Meth-A cells, but when these same cell populations were treated on a Sephadex G-10 column, the effector activity seen in Winn's assay disappeared. The effector cells responsible for tumor inhibition were therefore considered to be cytotoxic macrophages. However, the inhibitory effect of these cytotoxic macrophages in Winn's assay was not evident in either X ray (300 rad)-irradiated BALB/c mice or in nu/nu BALB/c mice. These results indicate that the antitumor activity of cytotoxic macrophages is associated with a sequential immune mechanism in which T cells may play an important role.     

Clinical effect of combined chemotherapy with UFT, MMC and OK-432 in recurring or advanced cancer of the digestive organs

The usefulness of UFTMO therapy (combined chemotherapy with UFT, MMC and OK-432) performed in 40 cases of recurring or advanced cancer of the digestive organs was investigated. According the response criteria by Koyama et al., of 40 eligible cases, the treatment was judged effective in 13, 2 CR and 11 PR cases with a response rate of 32.5%, while of the 35 complete cases, 2 CR and 9 PR cases made for 11 effective cases and a response rate of 31.4%. Side effects were observed in 58.3% of the 36 evaluated cases; of the subjective and objective side effects, however, none were serious enough to require cessation of administration, while stopping administration in the cases of abnormal laboratory findings resulted in rapid recovery. UFTMO therapy, therefore, is considered to be one of the beneficial treatments for recurring or advanced cancer of the digestive organs.     

Anti-tumor immunity induced by OK-432-derived DNA

We have tried to identify the effective components of OK-432, a Streptococcus-derived anti-cancer immunotherapeutic agent. In the current study, we investigated the effect of OK-432-derived DNA (OK-DNA) in augmenting anti-cancer immune response. Analysis of OK-DNA with the restriction enzymes Hpa II and Msp I revealed that OK-DNA contained unmethylated CpG motifs. OK-DNA induced Th1-type cytokines, such as IFN-gamma and IL-12, and augmented killer cell activities in vitro on human peripheral blood mononuclear cells, whereas the methylated OK-DNA did not. Cytokines were also produced by OK-DNA-stimulated splenocytes derived from wild-type mice but not from TLR9-deficient mice. In the in vivo study, a peritumoral administration of OK-DNA resulted in a significant inhibition of tumor growth in syngeneic tumor-bearing wild-type and TLR4-deficient mice but not in TLR9-deficient mice. Anti-tumor effect of OK-432 in TLR9-deficient mice was significantly but partially reduced as compared with that in wild-type mice, while the effect of OK-432 was almost completely eliminated in TLR4-deficient mice. These findings suggest that unmethylated CpG-DNA in OK-432 functions as an active component in OK-432-induced anti-cancer immunity via TLR9, at least in part.     

Induction of mouse peritoneal cytotoxic factor by OK-432. (3). Comparison between LPS and OK-432 as eliciting agents

By injection of OK-432, a cytotoxic factor was induced in peritoneal fluids of mice which had been primed with OK-432. Two-step stimulation (priming and eliciting) was always necessary to induce the cytotoxic factor. OK-432-primed mice did not produce soluble cytotoxic factor spontaneously and no cytotoxic activity was detected in the mice treated by a single injection of OK-432 as an eliciting agent. High doses of OK-432 were required to prime mice for the production of cytotoxic factor, whereas a small amount was enough to elicit it. Pathological studies were also conducted in order to clarify whether the mice were safe under the conditions in which PCF had been induced. Moderate liver damage was observed in the mice injected with OK-432 and LPS, whereas no histological change in the liver or spleen was observed in the mice treated with OK-432 alone. These results suggest that OK-432 is a good candidate as an inducer of cytotoxic factor in the peritoneal cavity.     

Immune hemolytic anemia associated with streptococcal preparation OK-432

In the course of administration of an immunopotentiator, streptococcal agent OK-432, a patient with lung cancer was found to have severe anemia and a strongly positive antiglobulin test. In the absence of the drug, eluates from the patient's erythrocytes reacted strongly by the indirect antiglobulin test with a panel of washed Group O erythrocyte samples. The eluated antibody was composed of monoclonal IgG1-lambda. Upon discontinuing the drug the patient's hemoglobin level increased slowly with concomitant normalization of the reticulocyte count. Although very rare, drug-induced immune hemolytic anemia should be included in the list of adverse effects of immunopotentiators. This is the first reported case of immune hemolytic anemia associated with OK-432.     

Experimental and clinical effect of hypertransfusion and OK-432 on granulocyte recovery

The experimental and clinical studies were carried out to alleviate the bone marrow suppression by antineoplastic drugs. Faster recovery of granulopoiesis was observed by pretreating recipients with RBC hypertransfusion and/or OK-432 (Picibanil, biological products of beta-streptococci.) In experimental mice, higher granulocyte counts could be maintained with hypertransfusion in the peripheral blood, and the recovery of granulopoietic series and CFU-S of bone marrow cells were found to be more rapid after cyclophosphamide administration. OK-432 also resulted in higher peripheral granulocyte, whereas the total nucleated cell counts and CFU-S were decreased in the bone marrow, suggesting sparing bone marrow granulocyte reserve and its migration to the peripheral blood. The mechanism of higher granulocyte count after hypertransfusion was not clearly explained but it was considered that erythroid suppression caused colateral flow of multipotential stem cells to granulopoiesis. The effect of both combinations was unexpectedly less significant in the recovery of granulopoiesis, but it was thought that the optimal time interval should be sought between pretreatment and the administration of anti-neoplastic agents. The clinical use of hyper transfusion and OK-432 also proved the alleviation of granulocytopenia, and rapid granulocyte recovery at the time of consolidation therapy among children with AML.     

Immunotherapy, with OK-432 for cases of nasopharyngeal carcinoma

The prognosis of nasopharyngeal carcinoma (NPC) is very poor, due to the extreme failure of the immuno-surveillance mechanism in such cases. As an immunotherapeutic agent, OK-432 was administered to NPC patients. Cases were divided into two groups. One was given OK-432 for less than 6 months (short-term group), while a long-term group, was treated with OK-432 for over 6 months. These two groups were evaluated for immunological activity and prognosis. As immunological parameters, the numbers of white blood cells and lymphocytes were counted, and delayed skin reactivities with Su-PS and PPD were determined. No influence on the number of white blood cells and lymphocytes could be observed in either of the two groups. However, the skin tests showed better improvement of immunity in the long-term group than in the short-term cases. Furthermore, these reactivities correlated well with the clinical status of NPC patients. As to the absolute number of lymphocytes, improvement in the long-term NPC group was delayed in comparison to that of another head and neck carcinoma group given long-term OK-432 treatment. The prognosis of the long-term administered group was apparently better than that of the short-term group. Long-term administration of OK-432 is therefore indispensable for the treatment of NPC cases, because of the extreme decline of their immunity.     

Study on adjuvant immunochemotherapy with long-term OK-432 administration in colorectal cancer

The effect of OK-432 on the survival of patients with colorectal cancer who had undergone surgical resection was studied. As postoperative chemotherapeutic agents, mitomycin C and tegafur were administered. The survival time in the OK-432 group was prolonged in comparison with that in the chemotherapy-alone group (control) among patients given curative resection, but not among those not given curative resection. The effect of OK-432 was especially significant in patients with Dukes C-Stage and Dukes C rectal cancer (p less than 0.05). Moreover, OK-432 significantly inhibited the recurrence of cancer at the original lesion in patients with Dukes C cancer who had received curative resection. These results indicate that long-term administration of OK-432 is effective as a postoperative adjuvant for curatively resected colorectal cancer, especially Dukes C cancer.     

Advanced Ovarian Cancer Treated by Intraperitoneal Immunotherapy With OK-432

Twelve patients with advanced ovarian cancer with ascites weretreated by intraperitoneal immunotherapy with OK-432 in combinationwith surgery and systemic immunochemotherapy. The median timefor total resolution of ascites after the OK-432 injection was7.1 days. In three of the four patients who died, it took morethan the average number of days for the ascites to disappear.The ascites index, the ratio of the ascitic fluid volumes beforeand after intraperitoneal immunotherapy, was significantly higherin the patients who died than in those who survived. The sideeffects of intraperitoneal administration of OK-432 varied,but were not serious and relief was not needed in most cases. Intraperitoneal injection of OK-432 was considered effectivein preventing the reappearance of ascites in these cases ofadvanced ovarian cancer, and the ascites index was believedto be related to the prognosis.     

Adjuvant immunochemotherapy with long-term OK-432 administration in colorectal cancer

A new clinical trial on immunochemotherapy as an adjuvant therapy of surgery for colorectal cancers was studied. Results were retrospectively evaluated against previous controls treated by chemotherapy alone. As an immunotherapeutic agent, a streptococcal preparation, OK-432 was used. The maintenance dosage of OK-432, was 5 KE once a week and was continued for at least 2 years after surgical resection. As chemotherapeutic agents, mitomycin C for 2 weeks postoperatively and tegafur for 1 year were administered. Delayed skin reactivity to SU-polysaccharide (SU-PS) extracted from Streptococcus pyogenes SU-strain and lymphoproliferative response to phytohemagglutinins (PHA) were significantly enhanced in the OK-432, immunochemotherapy group. Disease-free interval in the immunochemotherapy group (n = 49) was prolonged compared to that in the control group (n = 129), especially in the curative resection cases of Duke's C stage. There was statistical significance between the two groups (p less than 0.05). These results suggested that long-term administration of OK-432 after surgical resection of colorectal cancer was effective on growth inhibition of micrometastasis and could increase the postoperative survival rate.     

OK-432增强CPT-11抗肿瘤活性的实验研究

目的：探讨OK－432增强CPT－11抗肿瘤活性的机制。方法：将荷B16黑色素瘤的C57BL／6小鼠随机分为对照组、CPT－11治疗组、联合治疗组,于接种后第3天腹腔分别注射生理盐水、CPT－11,CPT－11加OK－432。观察肿瘤生长体积;测定各组脾细胞分泌的IL－2,IL－4,IL－6,IL－10,IL－12,IFN－γ,并在体外观察了SN－38对OK－432刺激这些细胞因子分泌的影响。结果：OK－432显著增强CPT－11对B16黑色素瘤生长的抑制（P＜0．05）;CPT－11在体内显著抑制小鼠脾细胞IL－12和IFN－γ的分泌,与OK－432联合使用显著增加小鼠脾细胞IL－2,IL－6,IL－12和IFN－γ的分泌（P＜0．05）;在体外SN－38对OK－432刺激的IL－6,IL－12分泌无显著影响（P＞0．05）。结论：OK－432在体内可通过刺激IL－12和Th1细胞因子的分泌,增强CPT－11的抗肿瘤活性。     

Intrapericardial OK-432 instillation for the management of malignant pericardial effusion

Ten patients with malignant pericardial effusion were treated with intrapericardial injection of OK-432 (penicillin-treated and heat-treated lyophilized powder of the substrain of Streptococcus pyogenes A3). After intrapericardial insertion of a catheter, a maximal volume of pericardial fluid was withdrawn with cytologic confirmation of malignancy. Five or 10 Klinische Einheit (KE) (KE is a unit used to express the strength of a preparation) of OK-432 diluted in 20 ml of saline was injected into the pericardial space in seven and three patients, respectively. It was repeated in case of reaccumulation. Seven patients were treated only once and the remaining three required a second treatment. Complete control of pericardial effusion was achieved in all patients for an average of 329 days (range, 54 to 790 days). Fever and chest pain were experienced in six and five patients, respectively, but were controlled with antipyretics. Two of three patients who received 10 KE of OK-432 experienced hypotension that was successfully controlled with vasopressor drugs with or without reaspiration of pericardial fluid. Rapid reactive reaccumulation of the pericardial fluid was thought to be a cause of hypotension. A follow-up computed tomography (CT) scan was performed in seven patients and a thickened pericardium was noticed in five; no patients had constrictive pericarditis. These results suggest that intrapericardial administration of 5 KE of OK-432 is an effective and safe treatment for malignant pericardial effusion.