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High mobility group box-1 (HMGB1), a representative damage-associated molecular patterns (DAMPs), has been reported to be involved in many inflammatory diseases. To validate HMGB1 as a target molecule of inflammatory diseases and to examine the effects of inhibition of HMGB1 on the diseases, we raised anti-HMGB1 monoclonal antibody (mAb) neutralizing extracellular HMGB1 and characterized it. We report the effects of anti-HMGB1 mAb on brain infarction, brain hemorrhage, brain trauma, epilepsy and neuropathic pain using animal models. In these wide range of disease conditions, we found a common event; the injury-induced translocation of HMGB1 from nuclei to extracellular space, especially in neurons. Released HMGB1 disrupt the integrity of blood-brain barrier (BBB) and increased the permeability of BBB, associated with inflammatory responses including the induction of pro-inflammatory cytokines. The intravenous injection of anti-HMGB1 mAb inhibited translocation of HMGB1, BBB disruption, expression of inflammatory molecules and improved neurological symptoms of different kinds of disease models. These results as a whole indicated that HMGB1 may be a very sensitive factor which is mobilized readily by different injurious insults to the brain and that HMGB1 release may be present most upstream of cascade of events triggering BBB disruption and brain inflammation. Thus, HMGB1 may be an excellent target for the treatment of above-mentioned diseases. Anti-HMGB1 mAb provide a novel and potential therapy for these severe disease conditions.

In the development of quality biopharmaceutical drugs, it is the level of knowledge gained, not the volume of data, which provides the basis for science-based submissions and their regulatory evaluation [International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) Q8 (R2)]. The identification of critical quality attributes (CQAs) is important as the initial step for quality by design (QbD) strategies. The impact of each potential critical attribute (pCQA) is assessed by using a systematic and scientific approach with respect to bioactivity, pharmacokinetics (PK)/pharmacodynamics (PD), immunogenicity, and safety of the biopharmaceutical drugs. The purpose of this section is to introduce a strategy for identifying CQA, and new analytical methods for CQA assessment.

CCR4 is one of the receptors for chemokines and is expressed on Th2, Th17, skin-homing T cells and regulatory T cells (Tregs). Activated Tregs highly express CCR4 and are induced into tumor environment by M2 macrophages-producing CCL22. Tregs show strong suppres- sive functions in cancer immunity, resulting in tumor progression. On the other hand, moga- mulizumab is a humanized anti-human CCR4 monoclonal antibody with a defucosylated Fe region which markedly enhances ADCC activity, and the antibody has been shown to effi- ciently deplete CCR4-positive cells. Therefore, a phase Ia clinical study of mogamulizumab was conducted in the treatment of solid cancers. Mogamulizumab almost depeleted activated Tregs in peripheral blood, and then re-activated CTL function with low grade of adverse events. However, no clinical responses were observed in ten patients. A phase lb study is now on-going, and further clinical studies may be needed in combination with other anti- tumor drugs.

Ebolaviruses, members of the family Filoviridae, cause severe hemorrhagic fever (Ebola virus disease: EVD) in humans and nonhuman primates with case fatality rates of up to 90%. No effective prophylaxis or treatment for EVD is yet commercially available. The latest out- break of EVD in West Africa has highlighted the urgent need for the development of thera- peutics and diagnostics. This outbreak has indeed accelerated efforts to develop antiviral countermeasures and unapproved vaccines, drugs, and diagnostics were used during the outbreak, some of which have undergone clinical trials. This article reviews recent advances on the development of therapeutics and diagnostics for EVD, particularly focusing on the util- ity of virus-specific monoclonal antibodies.

Background: As anti-hepatitis B surface antigen (anti-HBs) titers vary depending on the measurement methods, we compared two different methods to measure anti-HBs titers in sera and HBs monoclonal antibodies., Methods: The sera from 182 HB virus-resolved patients who were negative for HBsAg but positive for antiHB core protein (HBc) and/or anti-HBs were obtained. The measurement of anti-HBs was compared using either Lumipulse G1200 or Architect i2000SR. Six different monoclonal antibody (mAbs) clones isolated from healthy individuals inoculated with hepatitis B vaccine Bimmugen (genotype C) were used., Results: A statistically significant correlation in anti-HBs titers was found between the two methods tested (Y = 0.951X + 100.7, R = 0.813, p < 0.001), although anti-HBs titers in 72 samples (39.6%) measured by Architect were less than 50% of that by Lumipulse and 12 (6.6%) were opposite results. Measuring 2 mAbs with HBV neutralizing activity, the titers of the 116 antibody (1.0 μg/mL) were comparable (689.3 mIU/mL by Lumipulse and 440.7 mIU/mL by Architect), whereas those of the 478 antibody (1.0 μg/mL) were much lower by Architect than by Lumipulse (42.6 vs. 818.6 mIU/mL, respectively). Of four other mAbs without HBV neutralizing activity, equal titers were observed for one; two mAbs had less anti-HB titers by Architect; and one was below the cut-off index (< 5 mIU/mL)., Conclusions: Anti-HBs titers measured by Architect are likely to be lower than by Lumipulse, and the potential ability to detect the 478 antibody with neutralizing activity is low, indicating that Architect might underestimate anti-HBs titers. Future studies should standardize the anti-HBs titer measurement system.

Multiple myeloma (MM) remains mostly incurable despite the recent progress in the treatment strategy. One of novel fields for anti-MM therapeutic strategy is the development of immunotherapy using monoclonal antibodies (MoAbs) against myeloma-specific antigens. This article focuses on the basic and clinical aspects of several emerging and promising novel MoAbs for MM, such as elotuzumab which targets CS1 and daratumumab which targets CD38. Both antigens are highly expressed in more than 90% of MM patients, and the clinical trials have shown promising anti-MM effects, especially in combination with immunomodulatory agent lenalidomide. We also discuss the characteristics and the results of clinical trials of other MoAbs, such as tabalumab against B cell activating factor or dacetuzumab against CD40, being developed for MM.

A 58-year-old woman was confirmed as having multiple liver metastases after undergoing a high anterior resection for a sigmoid colon tumor. She was administered bevacizumab+FOLFOX as the first regimen and bevacizumab+FOLFIRI and S-1 and irinotecan (IRIS)therapy as the second regimen. During this treatment she also underwent hepatectomy 3 times and radiofrequency ablation once. She was administered panitumumab+irinotecan as the third regimen and, due to the presence of multiple pulmonary metastases, was subsequently considered to have had a partial response (PR). Because she subsequently developed progressive disease (PD), she received the fourth regimen as part of a clinical trial (TAS102) in another hospital. Cetuximab+irinotecan was administered as the fifth regimen after PD and the tumor was found to have reduced in size by 23%upon computed tomography (CT) 2 months later. Although stable disease (SD) was achieved, she was subsequently administered regorafenib for 8 months as a sixth regimen after the disease progressed a second time. In some cases of KRAS wild type metastatic colorectal cancer, re-challenging with an anti-epidermal growth factor receptor (EGFR) monoclonal antibody seems to be an effective strategy for reducing tumor mass.

We evaluated the performance of a newly-improved estradiol(E2) assay reagent (NEW LP-E2-N), which replaces murine monoclonal antibody in the present reagent (LP-E2-N) with sheep monoclonal antibody, since we had experienced discrepant E2 assay results between LP-E2-N and other commercially available E2 assay kits. Several examinations with the new assay reagent indicated a good performance in terms of the limit of quantity, reproducibility (within-run and between-day), dilution linearity, and influence of blood components except hemoglobin. Using analogues and/or metabolites of E2, low or no cross-reactivity has been shown in NEW LP-E2-N: 0.26% with 25 ng/mL of estrone (El), 0.14% with 100 ng/mL of estradiol-3-sulfate, 0.02% with 200 ng/mL of 17α-ethynylestradiol, and less than 0.001% with 100 ng/mL of estriol(E3), estra-17-glucuronide, and estramustine, respectively. Although discrepant results between NEW LP-E2-N and LP-E2-N were observed in 12 samples, including 9 cases under oral hormone therapy, data from these samples were similar to those using 2 commercially available E2 assay kits, Architect and Eclusys, suggesting that the NEW LP E2-N shows adequate clinical efficacy. A correlation study was performed with LP E2-N, Architect, and Eclusys using 149 serum samples obtained from patients and healthy volunteers, and the correlation results were as follows: r = 0.831, y = 0.98x + 40.6 against LP-E2-N, r = 0.991, y = 1.08x + 12.4 against Architect, r = 0.995, y = 0.80x - 3.7 against Eclusys. In conclusion, the NEW LP-E2-N reagent displayed a relatively favorable kit performance except for in the elevation of assay results with hemoglobin, as well as a low cross-reactivity with E2 analogues and/or metabolites.

Chronic hepatitis C virus (HCV) infection is a global public health issue because ～30% of HCV-carriers develop severe liver diseases including hepatic steatosis, cirrhosis, and hepatocellular carcinoma. Not only viral factors but also host/viral interactions are promising targets for antiviral preventive and therapeutic strategies. Recent studies showed that a tight junction protein claudin 1 is involved in HCV entry into host cells. Consistent with these studies, we isolated the several hepatic Huh7-derived cell clones defective in claudin 1 as HCV-resistant mutants, and cellular permissiveness to HCV was restored by expression of claudin 1 into these cell mutants. These results strongly suggest that claudin 1 is a promising target for antiviral therapy. We thus tried to isolate antibodies against extracellular domain of human claudin 1. Finally we established four mouse anti-claudin 1 monoclonal antibodies by using DNA immunization method and hybridoma screening with the above claudin 1-defective mutant. In the cell culture-infection system using Huh7.5.1 cells and HCV-JFH1 strain, these four antibodies efficiently inhibited infection by HCV in a dose-dependent manner, but do not affect tight junction localization of claudin 1 and cellular barrier function. These monoclonal antibodies targeting claudin 1 might be useful for preventing HCV infection, such as after liver transplantation, and also blocking viral spread in HCV-infected patients.

Herbal medicines have recently attracted much importance owing to the rising interest in their health benefits. Hence, further elucidation of the functions and mechanisms of these natural compounds is necessary. Our laboratory has established more than 30 kinds of monoclonal antibodies (MAbs) against bioactive natural compounds. Moreover, we have developed highly sensitive measurement systems for natural compounds, such as enzyme-linked immunosorbent assay (ELISA) and eastern blotting using MAbs. To expand the application of these MAbs to the functional analysis of natural compounds, we established a new approach for the isolation of the target compound from plant extracts using an immunoaffinity column conjugated with an anti-natural compound MAb. Through one-step purification using a MAb-conjugated immunoaffinity column, we have succeeded in preparing a knockout (KO) extract containing all components except the target compound, used as a hapten. Furthermore, we examined the pharmacological effects of the KO extract to identify the precise roles of the bioactive compound in the plant extract. To confirm another beneficial use of MAbs, we investigated the cellular localization and target molecules of natural compounds by immunocytochemistry (ICC) and Western blotting using MAbs. Our results demonstrated that MAbs clearly determined the cellular localization and target molecules of the natural compounds. These approaches may make it possible to determine the potential functions and target molecules of bioactive natural compounds in herbal medicines.

Traumatic brain injury (TBI) is one of the major causes of death and aftereffects in young individuals worldwide; however, efficient therapies for TBI are lacking at present. High mobility group box-1 (HMGB-1), which is recognized as a representative of danger-associated molecular patterns (DAMPs), plays an important role in triggering inflammatory responses in many types of diseases. We presented the involvement of HMGB-1 in TBI and evaluated the ability of intravenously administered neutralizing anti-HMGB-1 monoclonal antibody (mAb) to attenuate brain injury. Anti-HMGB-1 mAb may provide a novel and effective therapy for TBI by protecting against blood brain barrier disruption and reducing the inflammatory responses induced by HMGB-1.

Although the treatment of cancer with monoclonal antibodies has long been pursued, T cell-directed immunotherapy has met with limited success. Recently, much attention has been devoted to the blockade of PD-1 signaling to activate an immune response to cancer. PD-1, a protein expressed on T cells, is a member of the CD28 superfamily, and it transmits coinhibitory signals upon engagement with its ligands PD-L1 and PD-L2. Accumulating evidence suggests that the PD-1 system plays pivotal roles in the regulation of autoimmunity, transplantation immunity, infectious immunity, and tumor immunity. Because the interaction of PD-1 with its ligands occurs in the effector phase of killer T cell responses in peripheral blood, anti-PD-1 and anti-PD-L1 monoclonal antibodies are ideal as specific agents to augment T cell responses to tumors with fewer adverse events than with the inhibition of CTLA-4, because the interaction of CTLA-4 with its ligands occurs in the priming phase of T cell responses within lymph nodes. In recent phase I clinical trials, objective responses were observed in patients with melanoma, renal cell carcinoma, and non-small cell lung cancer who underwent immunotherapy with an anti-PD-1 monoclonal antibody. In addition, the antitumor activity of an anti-PD-L1 monoclonal antibody was observed in patients with melanoma, renal cell carcinoma, non-small cell lung cancer, and ovarian cancer. The next frontier of immunotherapy targeting the PD-1 axis is to define patient selection criteria and explore combination therapy with other therapeutic manipulations such as adoptive immunotherapies.

Ipilimumab(MDX-010, BMS-734016)is a fully human monoclonal immunoglobulin(IgG1k)specific for human cytotoxic T lymphocyte antigen 4(CTLA-4, CD152), which is expressed on a subset of activated T-cells as a negative regulator of T-cell activation. Blockade of CTLA-4 can potentiate a robust antitumor immune response and lead to long-term tumor regression. Moreover, 2 global phase 3 clinical studies(MDX010-20 and CA184-024)on ipilimumab have demonstrated an improved survival in patients with both untreated and treated advanced melanoma. Ipilimumab(Yervoy TM)has been approved for clinical use for the treatment of advanced melanoma in over 40 countries, including the United States(March 2011)and the European Union(July 2011), as the first agent to show overall survival benefit in patients with advanced melanoma. In Japan, several clinical trials for ipilimumab are ongoing in patients with conditions such as melanoma, non-small cell lung cancer, small-cell lung cancer, and gastric cancer. On the basis of experiences in both clinical development and clinical use of ipilimumab in more than 13,000 patients, several unique features of immunotherapy have been identified. In this article, clinical data on ipilimumab along with its unique effects and safety profile will be introduced, and some possible options for the further development of ipilimumab will be briefly discussed.

Antibodies have greatly contributed to the development of medical science and pharmacology, because of their high specificity. The cell fusion method has developed monoclonal antibodies (mAb) technology, such that massive amounts of mAb with a uniform structure can be produced. Although mAb have been produced against many proteins so far, the production of mAb against multi-pass transmembrane proteins, such as G-protein coupled receptor (GPCR) and various transporter proteins has been extremely difficult. The complicated structures, poorly extracellular regions, and high hydrophobicity of multiple-transmembrane proteins make it difficult to produce mAb against them. Production of mAb that recognize the extracellular region of living cells is thought to be important in determining the ability of a protein. Based on these findings, we tried to produce mAb against a multi-pass transmembrane transporter using green fluorescent protein (GFP)-fused full-length target proteins as immunogens. Furthermore, the immunizing method has proved to be important in generating functional mAb. We succeeded in producing functional mAb that react against the extracellular region of a 12-pass transmembrane transporter in a living cell. Based on this success, we began to produce mAb against seven-transmembrane GPCR. In this symposium, we report on the results of producing mAb against S1P receptors, a type of GPCR.

Monoclonal antibodies play important roles in medical oncology. The antibodies were designed as specific molecular targeting drugs and supposed to have less toxicity to normal cells compared to classical cytotoxic agents. Indeed, they do not have severe bone marrow suppression, nausea, or vomiting unlike cytotoxic drugs. On the other hand, clinicians often undergo characteristic adverse events we have never experienced before the appearance of the molecular targeting drugs. To fully utilize these powerful yet particular medicines, we have to be well aware of their severe or fatal adverse events and comprehend how to manage those toxic events. In this manuscript, important adverse events including infusion reaction, gastrointestinal perforation, cardiotoxicity, venous thromboembolism, and interstitial lung disease are subjects for discussion.

Monoclonal antibodies have become the most effective therapeutic modality for treating human cancer. Anti-cancer monoclonal antibodies can be targeted against cancer cells by several mechanisms. Even unconjugated antibodies show significant efficacy in the treatment of solid tumors and hematological malignancies. Immunoconjugates composed of antibodies conjugated to chemo-drugs, radioactive or toxins are powerful therapy for lymphomas and solid tumors. Moreover, immunomodulatory antibodies can promote the induction of anti-tumor immune responses by directly activating or inhibiting molecules of the immune system. Antibody structures now can be readily manipulated to facilitate selective interaction between the immune system and cancer cells, so that these reagents will become important components of the anti-neoplastic protocols of the future.

We investigated a case in our experience presenting false-positive for DUPAN-2 by IgM-human anti mouse antibody (HAMA). A female aged 40s has been treated in our hospital from 2003. Her serum level of DUPAN-2 in 2005 and 2006 were 110 U/mL and 140 U/mL respectively. While this level was increased to 770 U/ml in 2007, and kept in the higher level so far. Around years of 2007, no meaning changes was detected by radiological and laboratory tests, and there was no significant change in her clinical signs and symptoms and medications. The elevation of DUPAN-2 was thought as a false-positive phenomenon and the mechanism was investigated. As results, no dilution lineality was found, and absorption test showed a 95% reduction of DUPAN-2 levels not by IgG and IgA, but IgM-specific antiserum. Dithiothreitoldeacylation test with neuraminidase, and absorption test with HBR-1, IIR, and mouse IgM serum, it was suggested that the IgM with HAMA activity of the patient reacted with mouse monoclonal antibodies in reagents. Moreover, the HPLC-eluted fraction of DUPAN-2 of this patients was detected in the fraction of IgM, which as different from that of a control pancreatic cancer patient. Above these data, elevation of DUPAN-2 in this patient was a false positive phenomenon by IgM-HAMA reacting with mouse monoclonal antibodies in reagents. Although there are few reported cases of false positive phenomenon in DUPAN-2 measurement, we have to pay attention to such phenomenon when detecting an unlikely higher levels that could not be explained by clinical information.

Breast cancer is known to be associated with a high incidence of bone metastases. Recent advances in treatment for breast cancer have improved patient prognosis, including those with bone metastasis, highlighting the importance of treating bone metastasis to reduce incidence of skeletal complications and to improve patients' QOL. Currently, bisphosphonates(BP), which are recommended by domestic and international clinical practice guidelines, are commonly used for the treatment of bone metastasis. However, the outcomes of BP therapy leave room for improvement in regard to their efficacy, safety, and convenience. Prior studies have indicated that RANK ligand(RANKL), a cytokine mainly expressed in osteoblasts and bone marrow stromal cells, plays an important role in bone resorption by osteoclasts, which are key mediators in the formation and progression of bone metastasis. Denosumab is a fully human monoclonal anti-RANKL antibody which suppresses differentiation, activation, and survival of osteoclasts by inhibiting the binding of RANKL to its receptor, RANK. In a phase III clinical trial, denosumab significantly decreased the time-to-first and time-to-first-and-subsequent skeletal related events(SRE), compared with zoledronic acid in advanced breast cancer patients with bone metastases. Further more, denosumab was more effective than zoledronic acid in preventing the progression of bone pain and maintaining patients QOL. In the future, treatment of bone metastases for breast cancer patients is expected to evolve further with the introduction of denosumab, which is conveniently administered by subcutaneous injection.

Bone metastasis is relatively common in patients with breast, prostate, and lung cancer, and may cause severe bone pain, pathological fractures, hypercalcemia and other bone-related complications that drastically undermine quality of life. The currently-available treatments for bone metastasis are unsatisfactory in their effectiveness and outcome. Bone continuously undergoes remodeling through a repeated cycle of osteoclastic bone resorption and osteoblastic bone formation. When cancer cells metastasize to the bone, their growth is promoted under the influence of a variety of growth factors that are supplied from the bone as a consequence of osteoclastic bone resorption. This, in turn, causes an increased production of osteoclast- and osteoblast-stimulating cytokines in these cancer cells, leading to a further increase in bone remodeling. This vicious cycle between bone and metastatic cancer cells supports the development and progression of bone metastases. Accumulating evidence shows that RANKL, a cytokine expressed in osteoblasts/bone marrow stromal cells, plays an important role in the formation, activation, and survival of osteoclasts, which are key players in bone remodeling. RANKL is thus expected to inhibit bone metastasis by disrupting this vicious cycle, and is considered a rational new therapeutic intervention for bone metastasis. In animal models, inhibition with RANKL has been shown to prevent the development of bone metastases. Based on the promising results from these preclinical studies, a fully human monoclonal anti-RANKL antibody, denosumab, has been developed. In three phase III clinical trials of denosumab versus zoledronic acid(ZA)in advanced cancer patients with bone metastases, denosumab was superior or equal to ZA in delaying the time to the first on-study skeletal-related events(SRE). In conclusion, denosumab, a fully human monoclonal anti-RANKL antibody with a targeted mechanism of action, enables us to more easily and effectively treat bone metastases and manage cancer patients with bone metastases.

The epidermal growth factor receptor (EGFR), a receptor tyrosine kinase, triggers a downstream signaling cascade through areas such as the RAS-RAF-MAPK and PI3K-AKT pathways, which are involved in cell proliferation, survival and motility.Inhibiting EGFR activation has demonstrated significant promise as a molecular targeting therapy for various solid tumors. Two monoclonal antibodies (mAbs) targeting EGFR, cetuximab and panitumumab, are established to be new treatment options for metastatic colorectal cancer (mCRC). Among activating mutations in downstream of EGFR, the KRAS mutation, which is present in 40% of mCRC patients, has shown to be a predictive biomarker for resistance to anti-EGFR antibody therapy based on Caucasian studies. However, only a small proportion of patients achieved an objective response and benefit from anti-EGFR antibody, even among those with wild-type KRAS tumors. Other downstream factors in EGFR signaling are now being explored, such as the BRAF, PIK3CA, PTENgenes. Cetuximab, a chimeric immunoglobulin 1 (IgG1) monoclonal antibody, may also exert antitumor effects through antibody-dependent cell-mediated cytotoxicity (ADCC). ADCC is influenced by FCrR1 a-H131R and FC7RIKa-V158F polymorphisms. Additional analysis of BRAF, PIK3CA, PTEN and FcqR genes in KRAS wild-type patients could narrow down the selection of patients who are most likely to benefit from anti-EGFR antibody therapy.

Cetuximab is a monoclonal antibody that inhibits human epidermal growth factor receptor, and was approved for metastatic advanced colorectal cancer (mCRC) in 2008 in Japan. Evidences confirming the efficacy of cetuximab have been accumulated in western countries. As the first- and second-line therapy, cetuximab plus chemotherapy showed longer survival compared with chemotherapy alone. As a third-line chemotherapy, among various anti-cancer agents for mCRC, only cetuximab could exhibit survival benefits in monotherapy or combination therapy with irinotecan. Recent studies suggest that the status of KRAS mutation is a predictive marker in colorectal cancer patients treated with cetuximab, and these findings lead to personalized cancer treatment.

The use of cetuximab, a mouse chimeric immunoglobulin G1 monoclonal antibody, is approved as anti-epidermal growth factor receptor(EGFR)therapy for the treatment of metastatic colorectal cancer in Japan. Further, panitumumab, matuzumab, nimotuzumab and zalutumumab which also target EGFR, are currently under clinical development. Cetuximab is the first that has been developed as an anti-EGFR antibody. Approximately 30% of the protein which constructs the mouse chimeric antibodies is from mouse, which yields the possibility that the mouse chimeric antibodies induce host immune-reaction. After cetuximab, the humanized monoclonal antibodies such as matuzumab and nimotuzumab, and fully humanized monoclonal antibodies such as panitumumab and zalutumumab, have been developed. In this article, we will introduce the current status of development of these four anti-EGFR antibodies, by focusing on the individual clinical trials using each anti-EGFR antibody.

Unlabelled: Bevacizumab has been demonstrated to prolong survival in patients with metastatic colorectal cancer when used in combination with chemotherapy. We investigated the efficacy of chemotherapy administered in our general hospital for patients with metastatic colorectal cancer after the introduction of FOLFOX/FOLFIRI (+/-bevacizumab) therapy., Subjects and Methods: The subjects in this study were 34 patients diagnosed with metastatic colorectal cancer, who received either FOLFIRI (+/- bevacizumab) or mFOLFOX6 (+/-bevacizumab) therapy in this hospital. The subjects were divided into a bevacizumab combination regimen group (those who received the regimen as first-line treatment), and a non-bevacizumab combination regimen group (those who did not receive the regimen as first-line treatment). Comparisons were made with regard to anticancer efficacy, progression-free survival time, and overall survival time., Results: Comparison between the bevacizumab combination regimen group and the non-bevacizumab combination regimen group, revealed no significant difference due to the small number of relevant patients. However, the former showed a slight advantage over the latter in terms of anti-cancer efficacy and progression-free survival time., Conclusion: Bevacizumab is expected to contribute to the prolongation of survival in patients in our general hospital.

EGFR constitute an attractive target for tumor therapy, because it is a transmembrane receptor tyrosine kinase which is critically involved in tumorigenesis by stimulating cell proliferation and inhibiting apoptosis or other biological functions. The anti-tumor effects of targeted therapy by anti-EGFR monoclonal antibodies are mainly based on direct inhibition of the EGFR signal transduction pathway. In addition, monoclonal antibody has the potential advantage of recruiting immune effect or mechanisms to kill tumor cells. The pre-clinical and clinical data indicated that antibody-dependent cellular cytotoxicity (ADCC) contributes to tumor cell lysis by anti-EGFR monoclonal antibodies. Some polymorphisms in Fc gamma receptor genes have been shown to be a predictive marker for the efficacy of anti-EGFR antibodies. Continued research on the immunotherapeutic mechanisms of monoclonal antibodies will improve the efficacy of antibody-based targeted therapy for cancer.

Clinically, R-CHOP-like therapy plus radiation therapy is commonly performed for patients with limited stage diffuse large B-cell lymphoma. However, the efficacy and the safety of the management have not been evaluated properly. In particular, we have few definitive reports about patients with stage I DLBCL. This time we evaluated the effect of CHOP+/-R-like therapy plus radiation therapy, by analyzing 28 patients with stage I DLBCL, retrospectively. 15 patients were treated with the RCHOP-like therapy, and 13 received CHOP-like therapy combined with radiation therapy. A complete response was observed in all of the patients. With a median follow-up time of 14 months, 1-year progression-free survival (PFS) was 100%, and the 1-year overall survival (OS) was 100% for the patients receiving the R-CHOP-like therapy. With a median follow-up time of 68 months, 5-year PFS was 84. 6%, and 5-year OS was 100% for patients receiving the CHOP-like therapy. Since the followup time was not enough and the patient numbers were too few, the benefit of the addition of Rituximab to the CHOP therapy could not be clarified. We need to assess the safety and the efficacy of the combined modality therapy for patients with limited-stage DLBCL by a larger prospective study.

Cetuximab and panitumumab, both anti-epidermal growth factor receptor (EGFR) monoclonal antibodies (MAb), have demonstrated clinical activity in patients with colorectal cancer. They are well tolerated, but they involve various adverse events that are rare among cytotoxic agents. Typical adverse events associated with anti-EGFR MAbs include infusion reaction, skin toxicity, lung toxicity, and hypomagnesemia. It is necessary to recognize and manage adverse events promptly to continue treatments without drug discontinuation. This report details the adverse events and their management with anti-EGFR MAbs use.

For the patient in whom the Kras gene variation appears, cancer cell grow and continue to multiply even if blocked by EGFR inhibitor. A topping the offered figure effect of cetuximab is not accepted. The addition of cetuximab for treatment of metastatic colorectal cancer with Kras wild-type tumors is effective.

The revised version of the Japanese colorectal cancer treatment guidelines by the Japanese Society for Cancer of the Colon and Rectum published in July 2009 showed remarkable changes in the field of systemic chemotherapy compared with the 2005 edition. Bevacizumab was approved in 2004 in United States, in 2005 in Europe, and in 2007 in Japan. On the other hand, cetuximab was approved in 2004 in Europe and United States, and in July 2008 in Japan. Besides, capecitabine was approved in September 2009 in Japan for not only adjuvant chemotherapy but also unresectable advanced colorectal cancer. Thus, we had one more treatment option of capecitabine with Oxaliplatin as CapeOx (XELOX). Therefore, most of the standard chemotherapy regimens in Western countries then became available in Japan. There has been no major difference in the drug treatment strategy except for the approval of panitumumab in Europe and the US, but this was not true in Japan. Now KRAS testing is recommended, and the indication for cetuximab is limited to KRAS wild type. Cetuximab can not be administered as the first-line treatment in Japan because KRAS testing is not covered by health insurance. This article deals with the difference in the anti EGFR antibody indication between the Japanese and overseas guidelines.

The patient was a 73-year-old woman. Her chief complaints were abdominal pain and lower abdominal distension. After some examinations, we diagnosed pelvic tumor, bladder cancer (adenocarcinoma) and sigmoid colon cancer. We performed a first operation, but the pelvic tumor was firmly fixed anterior to the sacrum and right common pelvic artery. We judged it unresectable and performed tumor biopsy and ileostomy. The pathological findings were very similar to sigmoid colon cancer, so we diagnosed that the pelvic tumor was sigmoid colon cancer with extramural progression. Later, the patient was treated with three courses of FOLFOX4 and three courses of bevacizumab+FOLFOX4. After this chemotherapy, the pelvic tumor was reduced significantly, we considered resection possible and performed pelvic exenteration. She has had no recurrence for 6 months after second operation. This treatment appeared to be effective for unresectable primary colon cancer.

The case was a 69-year-old man with liver metastasis who had relapsed after the resection of rectal cancer. We treated the patient with FOLFIRI+bevacizumab(BV)after the failure of S-1+CPT-11, radiation, UFT/LV, FOLFOX and FOLFIRI regimen. The tumor marker level and accumulation of FDG-PET were rapidly decreased after initiating FOLFIRI+BV. There was no serious adverse event, and a remarkable improvement of QOL was observed. This case suggests that the BV combination regimen is useful even after the failure of FOLFOX and FOLFIRI regimen.

The incidence of pancreatic endocrine tumor (PET)accounts for 1 approximately 2% of total pancreatic tumors and 0. 4 approximately 1. 5% of autopsy cases, reflecting the recently increasing trend. According to WHO criteria(2004), PET is classified by the type of hormone produced by the tumor and its biological behavior. Together with the classical clinical images and hormone markers, 11C-5-HTP-Positron emission tomography, OctreoScan ([(111)In-DTPA0]octreotide)scintigram, SACI-test and IOUS are used for diagnosis. Surgery is the treatment of choice, if supposed to be curative and tolerable. In case of a well-differentiated endocrine tumor, with no indication of resection or IVR, somatostatin analog is another therapy showing stable disease status for a long period. Systemic chemotherapy, including 5-FU+streptozotocin, and streptozotocin+doxorubicin, are used in cases of well -differentiated endocrine carcinoma, and cisplatin+etoposide are applied for poorly-differentiated endocrine carcinoma (or small cell carcinoma). Recent studies focus on molecular target therapy including small molecules and monoclonal antibody, such as tyrosine kinase inhibitor, anti-VEGF antibody and moor inhibitor.

Background: Though irinotecan hydrochloride(CPT-11)was approved in Japan in 1994, there have been few reports since that evaluated the efficacy of CPT-11. The position of this agent in the treatment of patients with metastatic breast cancer(MBC)is not definite. In addition, no report has been published to date about CPT-11 and trastuzumab combination therapy., Purpose: To evaluate retrospectively the efficacy of CPT-11 and trastuzumab combination therapy as salvage treatment in patients with human epidermal growth factor receptor 2 (HER2)overexpressing MBC., Patients and Method: We examined ten cases who received this therapy against MBC since February 2002 till March 2007 in our hospital. Overall response rate, change of tumor markers, time to treatment failure (TTF), time to progression( TTP), overall survival (OS)after start of CPT-11 and adverse events were examined for efficacy and tolerability., Results: Median age was 57 (40-67)and median number of prior chemotherapies was 5(2-9). Though the overall response rate was 20%, some tumor reduction was observed totally in seven cases. CEA and CA15-3 were decreased in 78%(7/9)and 63%(5/8) of cases, respectively. Median TTF, TTP and OS were 3, 4, and 6 months, respectively. Adverse events included three cases of severe neutropenia, one of whom died of treatment-related sepsis. Slight diarrhea occurred in seven and severe nausea occurred in two cases. Dose modification of CPT-11 was necessary in 5 cases, and three discontinued CPT-11 administration, mainly due to easy fatigability, nausea and neutropenia. During the therapy, four cases were all inpatients, and 6 eventually became outpatients., Discussion: This therapy for patients with HER2 overexpressing metastatic cancer resistant to multi-agents demonstrated a good result in terms of antitumor effect. But high tolerability could not be documented by our experience with this therapy. It was supposed that the risk management with prediction of adverse events and preparation of better supportive therapy could make for the higher tolerability of this therapy for more effective clinical use.

Rituximab is commonly incorporated into CD20-positive B-cell lymphoma therapy to improve response and prognosis. With increasing use, resistance to rituximab is a continuing concern, but CD20 mutation as a cause of resistance has not previously been reported. Freshly collected lymphoma cells from 50 patients with previously untreated or relapsed/resistant non-Hodgkin's B-cell lymphomas(diffuse large B cell, n=22; follicular, n=7; mucosa associated lymphoid tissue, n=16; chronic lymphocytic leukemia, n=2; small lymphocytic lymphoma, n=1; lymphoplasmacytic, n =1; mantle cell lymphoma, n=1)were assessed for CD20 expression by flow cytometry, and CD20 gene sequencing was performed on extracted DNA. CD20 mutations were found in 11 of 50 patients(22.0%), and could be grouped as: C-terminal deletion(8.0%), early termination(10.0%), and extracellular domain(2.0%)or transmembrane domain (2.0%)mutations. The mean fluorescence intensity of CD20 on fresh lymphoma cells was significantly lower for the Cterminal deletion mutation(3.26; 95% CI: 0.09-6.89)compared with wild type(30.8; 95% CI: 22.4-39.2)(p<0.05). In contrast, early termination mutations did not show significant differences in CD20 expression compared with wild type (19.5; 95% CI: 10.7-28.4)(p>0.05). It is possible that C-terminal deletion mutations of CD20 may be related to relapse/resistance after rituximab therapy. These mutations should be examined in patients showing progression of disease after partial remission. Other mechanisms are also discussed.

We report a case of a 52-year-old female with multiple skin metastases from breast cancer who responded to S-1. She underwent surgery of her right breast in March 2000 and the left in June 2006. Multiple skin metastases were later detected. As there was no hormone sensitivity, chemotherapy was selected. We started treatment with administration of S-1 at 120mg/day(4 weeks)and trastuzumab. Thoracic CT at the end of the three courses revealed the disappearance of the multiple skin metastases. No major side effects were seen. It was concluded from these findings that S-1 could be safely and effectively administered to advanced breast cancer patients.

The prognosis of advanced colorectal cancer has been improved by introduction of molecular targeting agents, such as bevacizumab and cetuximab. Several clinical trials revealed a median survival time of more than 2 years. However, the main purpose of chemotherapy for advanced colorectal cancer has not been cure but prolongation of life to date. One explanation of the difficulty of cure is the inherent and/or acquired resistance to chemotherapeutic agents and molecular targeting agents. A better understanding of the mechanisms of inherent resistance will make it possible to identify predictive markers of responder or non-responder to select the optimal patients for a certain drug. Additionally, knowledge of the acquired mechanism will provide for the rational development of therapies that circumvent or overcome resistance. Although the mechanisms of the resistance of molecular targeting agents for colorectal cancer are still unclear, the inherent resistance to cetuximab or panitumumab by why of expression of KRAS mutation is the first breakthrough in this field. Further basic research and biomarker analysis of large clinical study are necessary to clarify the resistance to molecular targeting agents.

A sixties-man had complained of melena. Colonoscopy revealed type 2 tumor at rectum. Computed tomography (CT)demonstrated lymph node metastasis in front of sacrum and two low density areas which were suspected metastases in the liver. The patient was diagnosed stageIV rectal cancer and resected primary focus and lymph node metastasis.[ Ra-RS, ant, type 2, moderately differentiated adenocarcinoma, ly1, v3, pSE, pN2, sH1(Grade C), sP0, pM1(No. 270)]without liver resection. It was due to high level of CEA and remote lymph node metastasis. The patient was treated with mFOLFOX6 and bevacizumab after the operation. The level of CEA decreased to normal level and CT revealed a partial response after 4 cycles of systemic chemotherapy. Liver resection was performed safely. Histological response was Grade 2 at liver metastases.

We report an 83-year-old female with axillary node metastasis from breast cancer who responded to trastuzumab/ capecitabine combination therapy. In April 2007, she underwent surgery and at the same time axillary node metastasis was detected. As there was no hormone sensitivity, chemotherapy was selected, and administration of capecitabine at 1,800 mg/day(2 divided doses)and trastuzumab was initiated. Thoracic CT at the end of the six courses revealed the disappearance of the axillary node metastasis. No major side effects were produced. It was concluded from these findings that trastuzumab/capecitabine combination therapy could be safely and effectively administered to elderly advanced breast cancer patients.

Vascular endothelial growth factor(VEGF)is a key mediator in angiogenesis, an essential step in tumor growth and metastasis. Elevated levels of VEGF observed in solid tumors are correlated with worse clinical outcomes. Research has therefore focused on developing agents that target VEGF to inhibit tumor growth. Bevacizumab(Avastin), a recombinant humanized monoclonal antibody targeting VEGF. Bevacizumab with conventional chemotherapy has shown antitumor activity in several cancer types with acceptable toxicity. In April 2007, the Japanese Ministry of Health, Labour and Welfare approved bevacizumab for patients with unresectable colorectal cancer.

A 63-year-old woman had a 12 cm tumor on her right breast with broad skin redness, satellite lesions and 8 cm ipsilateral lymph nodes swelling(T4bN2aM0, Stage IIIB). Core needle biopsy and immunohistochemistry of breast tumor showed invasive ductal carcinoma with negative hormone receptor(ER-, PgR-)and overexpression of HER2 (HercepTest 3+). She was treated with weekly paclitaxel(80 mg/m(2), 4 administrations with a week rest)and a com- bination with weekly trastuzumab(initially 4 mg/kg followed by 2 mg/kg every week, totally 11 administrations). After 3courses of administration, the breast tumor, skin redness and axillary swelling were completely disappeared(clinical complete response), then mastectomy with axillary dissection was performed. Histopathology of the breast and lymph nodes showed complete disappear of invasive cancer cells with only 2x1 mm residue of ductal component(pCR, grade 3, DC+). We conclude that the combination of weekly paclitaxel and trastuzumab is a promising neoadjuvant therapy regimen for HER2 positive, ER-negative breast cancer.

The patient was a 73-year-old female with sigmoid colon cancer, who underwent resection of sigmoid colon cancer and liver metastasis. She was treated with 5-fluorouracil and levofolinate calcium(sLV5FU2)plus bevacizumab(BV) for advanced colorectal cancer. She was treated with angiotensin II receptor blocker(ARB)because hersystolic blood pressure was 200 mmHg and her diastolic blood pressure 100 mmHg after five courses of BV therapy. As a result, her blood pressure was controlled. It was possible to administer BV. Therefore, ARB may be the preferred antihypertensive agent in the management of BV-induced hypertension.

Vascular Endothelial Growth Factor (VEGF) plays an important role in proliferation of cancer cells, angiogenesis and vascular permeability in peritoneal dissemination. Now we investigated the efficacy of anti-VEGF therapy in peritoneal dissemination of gastric cancer considering of pharmacokinetics. We investigated pharmacokinetics of bevacizumab, humanized anti-VEGF monoclonal antibody in subcutaneous xenograft and peritoneal dissemination model of OCUM-2MD3, highly peritoneal-seeding cell line of human scirrhous gastric carcinoma. In the group of intraperitoneal administration, bevacizumab remains in the intraperitoneal cavity and transits slowly to blood pool. On the other hand, in the group of subcutaneous administration, bevacizumab transits faster to blood pool and is accumulated in subcutaneous tumors and peritoneal nodules. Bevacizumab administered intraperitoneally seemed to neutralize intraperitoneal VEGF. Bevacizumab administered in subcutaneous seemed to act on peritoneal wall and control vascular permeability. Systemic administration of anti-VEGF agent will be a potential therapy in peritoneal dissemination of gastric cancer.

We report a long-term survival case of metastatic breast cancer treated with trastuzumab and paclitaxel. The patient was a 50-year-old female. She underwent left mastectomy with axillary lymphadenectomy for advanced breast cancer with liver metastases. Histological examination showed papillotublar carcinoma, f, T3, ly3, v2, N2 (24/34), histological grade III, ER (-), PgR (-), HER2 (3+). Trastuzumab and paclitaxel was performed. One year after the operation, any lesions of liver were not detected by CT and she was diagnosed as a complete response. No evidence of viable lesions has been found for 7 years. Breast cancer with liver metastases often could be a life threatening, and therefore an optimal chemotherapy should be applied as soon as possible. Trastuzumab and paclitaxel would be one of the optimal chemotherapies for HER2 positive breast cancer.

Bevacizumab, a humanized monoclonal antibody to VEGF for advanced recurrent colorectal cancer, has been known for complications of gastrointestinal perforation, hemorrhage, thromboembolism and proteinuria, as adverse effects. These findings must be taken care as well as adverse drug reactions (ADR) caused by combination chemotherapy. We here in present a clinical experience in treatment with bevacizumab for unresectable colorectal cancer. Six patients treated with bevacizumab for over two courses until April 2008 were analyzed for this study. PR was obtained in one case with mFOLFOX6. Even though, grade 3 neutropenia was observed in only one case with FOLFIRI, the other cases had grade 2 or less in ADR. In addition, there were no any specific ADRs related with bevacizumab, so we concluded that combination chemotherapy for advanced recurrent colorectal cancer with bevacizumab was well-tolerated.

Monoclonal antibody treatment, which is one of the most promising molecular targeting cancer therapies, has recently become indispensable for the treatment of cancer due to its effectiveness and fewer side effects. Infusion reactions, which are similar to hypersensitivity or allergic reactions, are the generic term for the acute characteristic harmful reactions commonly associated with monoclonal antibody treatment. Those typically occur within the first 2 hours of the first infusion and are generally mild-to-moderate reactions which can be managed by either temporary interruption of infusion or administration of supportive care including corticosteroids, oxygen, or intravenous fluids. However, there are sometimes severe or life-threatening reactions, thus indicating the importance of closely observing the patient following the monoclonal antibody treatment. It is quite important that the entire medical staff understands the practical information regarding the timing and prevention or management of infusion reactions. In addition, it is also necessary to establish a system for prompt management of infusion reactions. Furthermore, sufficient information must be provided to the patient regarding infusion reactions.

Autopsy cases who had received Abeta vaccine showed clearance of senile plaques, and beneficial effect was shown in patients who had high antibody titers to amyloid plaques. Thus, vaccination is widely accepted as promising therapy for Alzheimer disease. Since active immunization induced autoimmune-like meningoencephalitis, pharmaceutical companies are interested in passive immunization using monoclonal antibodies to Abeta. However, due to immunization burden and economical issues, safe active immunization is still attractive. We developed oral or nasal Abeta vaccine using viral vectors and Abeta cDNA with a signal peptide. These vaccine reduced amyloid burden and improved cognitive functions in mice. We believe this vaccine is useful for prevention and treatment of Alzheimer disease.

The treatment of metastatic colorectal cancer (mCRC) has developed significantly over the past 10 years. For nearly 40 years, the fluoropyrimidine 5-fluorouracil (5-FU) was the only active agent used for advanced metastatic disease. However, since the 1990s, the chemotherapy treatment options for patients with mCRC have been greatly facilitated with the introduction of several new cytotoxic agents. In particular, combination regimens that incorporate infusional schedules of 5-FU in combination with oxaliplatin (FOLFOX) and/or irinotecan (FOLFIRI) have significantly improved clinical efficacy as related to overall response rates, time to tumor progression, and median overall survival. More recently, monoclonal antibodies such as bevacizumab, cetuximab, and panitumumab have become available for use in mCRC treatment in combination with cytotoxic agents and as monotherapies in Western countries. The addition of these target agents to the mCRC treatment armamentarium has resulted in more therapeutic options and improved treatment outcomes for patients. Currently, bevacizumab is the only target drug that is available for mCRC in Japan. In this article we review various treatment options, including cytotoxic and targeted agents, currently available for patients with mCRC in Japan and Western countries.

Gemtuzumab Ozogamicin (GO) targets leukemia cells expressing CD33 by means of a monoclonal antibody conjugated to a cytotoxic agent, calicheamicin. GO has been approved in Japan as monotherapy for the treatment of patients with relapsed/refractory acute myeloid leukemia (AML)since 2005. GO administered as a single agent has resulted in overall response rates of about 30% in previously relapsed adult AML. Preliminary data indicate a potential role for GO also as a component of induction or consolidation regimen. Although caution is advised when administering GO within 115 days of a stem cell transplantation (SCT) procedure because of veno-occlusive disease, recent clinical studies overseas suggest that GO can be integrated into reduced-intensity conditioning therapy before allogeneic SCT in patients with relapsed AML. In order to reduce toxicity and improve efficacy, its optimal dose and schedule should be defined by large clinical trials.