Kyorin Pharmaceutical Research Articles

1696. Urinary tract infection due to Aerococcus urinae in children

Abstract Background Aerococcus urinae is a rare Gram-positive and catalase-negative coccus. A. urinae is primarily associated with urinary tract infection (UTI). It is responsible for urosepsis and occasionally complicated diseases, including endocarditis and osteomyelitis. The clinical presentations of adult patients with A. urinae have been described. However, there are very few reports in children. The clinical course of UTI due to A. urinae and the associated risk factors remain unclear. The aim of this study was to describe the clinical characteristics of UTI due to A. urinae in children. Methods We conducted a retrospective observational study at a tertiary children’s hospital in Tokyo, Japan. We included pediatric patients (< 18 years old) who had positive urine cultures for A. urinae (≥ 104 CFU/HPF) between June 1, 2006, and May 31, 2022. Information regarding clinical characteristics, including age, sex, underlying disease, laboratory test results, antibiotic susceptibility of A. urinae, treatment, and outcomes were collected from the electronic medical records. Results A total of 22,769 urine cultures were submitted during the study period, of which A. urinae was isolated in 35 (0.15%). We found 17 patients who were positive for A. urinae with greater than 104 CFU/HPF. Among them, 13 patients (76.5%) were male and 16 (94.1%) had underlying bladder dysfunction. The main clinical symptoms were fever (n=6, 35.3%), urine odor (n=4, 23.5%), and nausea (n=2, 11.8%). Five patients had diagnosed pyelonephritis due to A. urinae. All five patients were male, and the median age at presentation was 4.6 years (range 2.1-11.0 years). Four of them had underlying bladder dysfunction. All bacterial isolates were susceptible to the majority of antibiotics, including penicillin G, sulfamethoxazole, or ciprofloxacin. All patients had favorable outcomes following 10-14 days of ampicillin/amoxicillin-based antimicrobial therapy. Conclusion A. urinae was isolated in urine cultures primarily in male patients with underlying bladder dysfunction. All of the outcomes of antibiotic treatment were favorable. Disclosures Kensuke Shoji, MD, PhD, AstraZeneca K.K.: Honoraria|Gilead Sciences, Inc.: Honoraria|KYORIN Pharmaceutical Co.,Ltd.: Honoraria|Meiji Seika Pharma Co., Ltd.: Honoraria|Nippon Becton Dickinson Company, Ltd.: Honoraria|Novartis Pharma Co., Ltd.: Honoraria|Viatrs, Inc: Honoraria chikara Ogimi, MD, bioMerieux Japan Ltd.: Honoraria|Horiba: Honoraria|Pfizer: Honoraria

631-P: Development of Noninvasive Diabetes Management Method Using Glycated Albumin in Tears

Aims/Background: Several clinical trials have demonstrated that strict glycemic control is important to minimize the risk of diabetic complications. Although frequent measurement of blood glucose levels by the invasive finger prick method is useful to achieve strict glycemic control, this method is invasive and burdensome to the patients. Glycated albumin (GA) is a biomarker which reflects the glycemic control status over the previous 2 weeks. We demonstrated GA levels in tears correlated those in blood. We hypothesized that regular GA tests in tears caused behavior change and improved glycemic control. Methods: Participants with type 2 diabetes were recruited and randomly divided to two groups. One group (intervention group) was performed tear GA tests every 2 weeks and reported the results, while the other group (control group) was not. After 8 weeks of intervention, the two groups were compared. GA levels in tears were measured by Liquid Chromatography / Mass Spectrometry. Results: We enrolled 56 participants to the study. Compared with the control group, the intervention group showed improvement in fasting blood glucose, HbA1c, GA, non-HDL cholesterol. Conclusions: Levels of GA, as a diabetes-related biomarker, in tears can be measured noninvasively. Regular measurement of the GA levels in tears could allow smoother management of diabetes mellitus and reduce the risk of complications noninvasively. Disclosure M.Aihara: None. N.Kubota: None. T.Yamauchi: Other Relationship; Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Company Limited, Novo Nordisk, Astellas Pharma Inc., AstraZeneca, Ono Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim, Kyowa Kirin Co., Ltd., Kowa Company, Ltd., Merck & Co., Inc., Sanofi, Sanwa Kagaku Kenkyusho, Shionogi & Co., Ltd., Daiichi Sankyo, Taiho Pharmaceutical Co. Ltd., Tosoh Corporation, Eli Lilly Japan K.K., Kissei Pharmaceutical Co., Ltd., Sumitomo Dainippon Pharma Co., Ltd., NTT Docomo Inc., Aero Switch, Asahi Mutual Life Insurance Company, NIPRO Medical Corporation, Abbott Japan Co., Ltd., Bayer Inc., Kyorin Pharmaceutical Company, Limited.

Signal Detection Study Focusing on Differences in the Drug Delivery System of Oral 5-Aminosalicylate for Inflammatory Bowel Disease Using the Japanese Pharmacovigilance Database

Although 5-Aminosalicylate (5-ASA) has been shown to act on the local mucosa, when 5-ASA is orally administered, most of it is absorbed in the upper gastrointestinal tract and does not reach the large intestine, where lesions are present. Therefore, different drug delivery systems have been developed for each oral 5-ASA formulation. Currently, the oral 5-ASA formulation approved in Japan is salazosulfapyridine (SALAZOPYRIN®; Pfizer Japan Inc.: Tokyo, Japan), in which 5-ASA and sulfapyridine are azo-bonded. In addition, there are several 5-ASA release formulations, including ASACOL®; ZERIA Pharmaceutical Co., Ltd.: Tokyo, Japan (delayed release formulation dependent on pH), PENTASA®; KYORIN Pharmaceutical Co., Ltd.: Tokyo, Japan (delayed release formulation dependent on time), and LIALDA®; MOCHIDA Pharmaceutical Co., Ltd.: Tokyo, Japan (delayed release formulation dependent on pH and time). Adverse events may occur because of differences in the drug delivery systems of these products. In this study, we focused on the adverse events of different 5-ASA formulations and investigated differences in the detection of safety signals for each 5-ASA formulation using disproportionality analysis. There were 15 adverse events detected only with SALAZOPYRIN®. On the other hand, ASACOL®, PENTASA®, and LIALDA® have different drug delivery systems. Although the detected signal intensities varied, the detected adverse events were not significantly different. These findings provide important insights, which should be considered by physicians during treatment selection and drug manufacturers during drug development.

134. KRP-A218, an Orally Active and Selective PI4KB Inhibitor with Broad-Spectrum Anti-Rhinovirus Activity, Has Potent Therapeutic Antiviral Activity In vivo

BackgroundRhinovirus (RV) is a major respiratory virus that poses a threat to immunocompromised people and those with underlying disease. However, there are no approved therapies. Moreover, RV infection cannot be prevented by a vaccine because there are over 100 serotypes. Here we report the pharmacological profile of a novel small-molecule host-targeted antiviral (HTA), KRP-A218 (A218). A highly potent and selective inhibitor of phosphatidylinositol 4 kinase beta (PI4KB), a key host factor of RV replication, A218 is undergoing clinical study.Methods in vitro antiviral activities of A218 and Vapendavir (Vap), a virus-targeted antiviral, were examined by inhibition of CPE, viral load, or replication. in vivo antiviral activity and pathological analysis of A218 were examined in coxsackievirus B3 (CVB3; belong to the genus enterovirus as with RV)-infected mice as a surrogate model of RV infection as CVB3, unlike RV, replicates very well in both mouse and human tissue. Daily oral dosing of A218 (1-10 mg/kg) was started 2 days post intraperitoneal infection with CVB3. Tissue viral load, pancreas pathological change at 4 days post infection, and survival rate up to 14 days were evaluated. PI4KB heterozygous kinase-dead mice (PI4KB KD) were established by a CRISPR-Cas9 system. Viral load and survival rate following viral infection were evaluated in these mice.ResultsA218 showed broad antiviral activity for RV and enteroviruses (Table) and has a higher barrier to drug resistance than Vap. These results are consistent with expectations for HTAs. Repeated dosing of A218 starting 2 days post infection decreased viral load and improved acute pancreatitis, accompanied by decrease of inflammatory and pancreatitis markers in plasma. Moreover, therapeutic dosing of A218 improved survival rate in a CVB3-infected lethal mouse model (Figure). These results show the first evidence that a PI4KB inhibitor has potent therapeutic efficacy in a severe viral infection model. Similar effects were observed in PI4KB KD, supporting the on-target effect of A218.Table. Antiviral activity of A218 and Vap against RV/EV infection Figure. Therapeutic effect of A218 on survival rate in CVB3-infected miceConclusionA218 is a promising therapeutic agent for improving the exacerbation of pathological conditions caused by RV infection. Nonclinical package including GLP-Tox also supports the ongoing first-in-human study of A218.Disclosures Toshiyuki Matsui, MPharm, Kyorin pharmaceutical Co., LTD (Employee) Motomichi Fujita, PhD, Kyorin Pharmaceutical Co., Ltd. (Employee) Yuji Ishibashi, PhD, Kyorin Pharmaceutical co., ltd. (Employee) Tyzoon Nomanbhoy, PhD, ActivX Biosciences (Other Financial or Material Support, Full time employee of ActivX, a wholly owned subsidiary of Kyorin Pharmaceuticals) Jonathan S. Rosenblum, PhD, ActivX Biosciences (Employee) Michiaki Nagasawa, PhD, Kyorin Pharmaceutical Co., Ltd (Employee)

Efficacy and Safety of Carboplatin With Nab-Paclitaxel Versus Docetaxel in Elderly Patients with Squamous-Cell Lung Cancer (CAPITAL): A Randomized, Multicenter, Open-Label, Phase III Trial

Efficacy and Safety of Carboplatin With Nab-Paclitaxel Versus Docetaxel in Elderly Patients with Squamous-Cell Lung Cancer (CAPITAL): A Randomized, Multicenter, Open-Label, Phase III Trial

Evaluation of the basic assay performance of the GeneSoc® rapid PCR testing system for detection of severe acute respiratory syndrome coronavirus 2

In the ongoing coronavirus disease 2019 (COVID-19) pandemic, PCR has been widely used for screening patients displaying relevant symptoms. The rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) enables prompt diagnosis and the implementation of proper precautionary and isolation measures for the patient. In the present study, we aimed to evaluate the basic assay performance of an innovative PCR system, GeneSoC® (Kyorin Pharmaceutical Co. Ltd., Tokyo, Japan). A total of 1,445 clinical samples were submitted to the clinical laboratory, including confirmed or suspected cases of COVID-19, from February 13 to August 31. Specimen types included nasopharyngeal swabs. The sampling was performed several times for each patient every 2-7 days. Using this system, sequences specific for SARS-CoV-2 RNA could be detected in a sample within 10-15 min using the microfluidic thermal cycling technology. Analytical sensitivity studies showed that GeneSoC® could detect the target sequence of the viral envelope and RNA-dependent RNA-polymerase (RdRp) genes at 5 and 10 copies/μL, respectively. The precision of the GeneSoC® measurements using clinical isolates of the virus at a concentration of 103 copies/μL was favorable for both the genes; within-run repeatability and between-run reproducibility coefficient of variation values were less than 3% and 2%, respectively; and the reproducibility of inter-detection units was less than 5%. Method comparison by LightCycler® 480 showed the positive and negative agreement to be 100% [(174/174) and (1271/1271), respectively]. GeneSoC® proved to be a rapid and reliable detection system for the prompt diagnosis of symptomatic COVID-19 patients and could help reduce the spread of infections and facilitate more rapid treatment of infected patients.

Discontinuation of Infliximab Increases the 1-Year Relapse Rate of Ulcerative Colitis in Remission - A Prospective, Multicentre, Randomised Controlled Trial

Background and Aims: Anti-tumour necrosis factor (TNF)- a agents are the mainstay of long-term treatment for refractory ulcerative colitis (UC). However, there has been no prospective randomised controlled trial evaluating whether anti-TNFa agents can be discontinued in UC patients in remission. Methods: This prospective, multicentre, randomised controlled trial enrolled UC patients from 23 specialist centres treated with infliximab (IFX) who were in remission. Patients who were confirmed to be: 1) in remission for > 6 months, 2) steroid-free, and 3) with a Mayo endoscopic subscore (MES) of 0 or 1, were randomised into two groups (continued IFX or discontinued IFX) in a 1:1 ratio stratified according to the use of immunomodulators and MES, using computer-generated stratified randomisation. The remission rate at week 48 in the full analysis set (FAS) was the primary endpoint in this trial. As secondary objectives, risk factors for relapse, and efficacy of re-treatment with IFX after relapse were also evaluated. Findings: A total of 95 patients were randomised, and 92 were included in the FAS. The remission rate at week 48 was 80.4% (n = 37/46; 95% CI: 66.1% to 90.6%) and 54.3% (n = 25/46; 95% CI: 39.0% to 69.1%) in the IFX-continued and IFX-discontinued groups, respectively (between-group difference: 26.1% [7.7% to 44.5%], p = 0.008). C-reactive protein and histological severity index were associated with future non-remission. Neither clinical nor endoscopic remission was predictive of remission. Retreatment with IFX upon relapse was safe, and two-thirds of patients achieved remission in 8 weeks. Interpretation: Discontinuing IFX resulted in a significantly increased relapse rate at 1 year. Residual subtle inflammation, as estimated based on CRP and histological inflammation seemed to be predictive of future relapse. Trial Registration:This study was registered at the University Hospital Medical Information Network Center Trials Registry (UMI12092). Funding Statement: This study was supported in part by a Grant-in-Aid for the Intractable Disease Project of the Ministry of Health, Labour and Welfare of Japan, and Mitsubishi- Tanabe Pharmaceuticals. Co. Declaration of Interests: Dr. Nakamura reports grants and personal fees from Mitsubishi Tanabe Pharma Corporation, grants and personal fees from EA Pharamaceutical Co Ltd, grants and personal fees from AbbVie Inc, personal fees from Janssen Pharmaceutical Co Ltd, grants and personal fees from Mochida Pharamaceutical Co Ltd, personal fees from Takeda Pharmaceutical Co Ltd, grants from Kyorin Holdings Inc, grants from JIMRO Co Ltd, grants from ZERIA Pharmaceutical Co Ltd, outside the submitted work. Dr. Motoya reports grants from Takeda pharmaceutical Co. Ltd, grants from Mitsubishi Tanabe Pharma Coporation, grants from Janssen Pharma coporation, during the conduct of the study. Dr. Hisamatsu reports grants and personal fees from EA pharma Co. Ltd., grants and personal fees from AbbVie GK, personal fees from Celgene K.K., personal fees from Janssen Pharmaceutical K.K., grants and personal fees from Pfizer Inc., grants and personal fees from Mitsubishi Tanabe Pharma Corporation, grants and personal fees from Kyorin Pharmaceutical Co. Ltd., grants and personal fees from JIMRO Co.Ltd, grants and personal fees from Mochida Pharmaceutical Co., Ltd, personal fees from Nichi-lko Pharmaceutical Co., Ltd., grants from Daiichi-Sankyo, grants and personal fees from Takeda Pharmaceutical Co. Ltd., outside the submitted work. Dr. Abe reports personal fees from EA Pharma Co.,LTD., personal fees from Kissei Pharmaceutical Co.,LTD., personal fees from Mochida Pharmaceutical Co.,LTD., personal fees from Mitsubishi Tanabe Pharma Corporation, outside the submitted work. Dr. Hibi reports personal fees from Aspen Japan K.K, grants and personal fees from Abbvie GK, personal fees from Ferring, personal fees from Gilead Sciences, personal fees from Janssen, grants and personal fees from JIMRO, personal fees from Kisse Pharmaceutical, grants and personal fees from Mitsubishi-Tanabe Pharma, grants and personal fees from Mochida Pharmaceutical, personal fees from Nippon Kayaku, personal fees from Pfizer, grants and personal fees from Takeda Pharmaceutical, grants and personal fees from Zeria Pharmaceutical, personal fees from Bristol-Myaers Squibb, personal fees from Celltrion, personal fees from EA Pharma, personal fees from Eli Lilly, grants and personal fees from kyorin, personal fees from Nichi-Iko Pharmaceutical, grants from Otuska Holdings, outside the submitted work. Ethics Approval Statement: The protocol was approved by the institutional review board of each participating institution. Written informed consent was obtained from all patients included in the study.

Exploratory Study of the Effectiveness of Granulocyte and Monocyte Adsorptive Apheresis Before Initiation of Steroids in Patients with Active Ulcerative Colitis During the COVID-19 Outbreak (EXPECT Study): A Multicenter Prospective Clinical Trial

Background: During the COVID-19 pandemic, safe treatment of inflammatory bowel disease is required. Granulocyte and monocyte adsorptive apheresis (GMA) has been used for therapy of steroid-dependent/refractory ulcerative colitis (UC) with few adverse events. Therefore, elucidating the effectiveness of GMA without steroids is important in the current situation. The aim of this study was to investigate the effectiveness of GMA in active UC patients who are not receiving steroids. Methods: A single-arm, open-label, and multicenter prospective clinical trial was conducted at ten facilities in Japan.UC patients who had insufficient responses to 5-aminosalicylic acid without steroids received GMA twice a week for five consecutive weeks. The primary and secondary endpoints were the induction of a response and endoscopic mucosal healing in the patients who achieved a response. Findings: A total of 67 patients were enrolled. The response rate of all patients who received GMA ten times was 58·2%. Of the 39 patients who achieved a response, 74·4% achieved endoscopically confirmed mucosal healing. Both the median Mayo score and ulcerative colitis endoscopic index of severity score were significantly decreased in the patients who achieved a response. The response rate of the patients who were suffering from their first episode was significantly higher than that of those with relapse-remitting UC (80·0% [16/20] vs. 48·9% [23/47], p=0·029). Undergoing the first episode of UC (odds ratio [OR]: 4·952, 95% confidence interval [CI]: 1·292-18·981, p>0.001) was defined as an independent predictor of a response. Interpretation: This study strongly suggest that GMA shows promising effectiveness in inducing remission in active UC patients who are not receiving steroid treatment. We expect that GMA can be made available during the COVID-19 pandemic. Trial Registration: UMIN registration No. 000013702 Funding Statement: Health and Labor Sciences Research Grants from the Ministry of Health, Labor and Welfare of Japan (Investigation and Research for intractable Inflammatory Bowel Disease to HN). Declaration of Interests: MM reports personal fees from Janssen Pharmaceutical K.K., and commercial research funding from AbbVie GK. and Nippon Kayaku Co., Ltd. SN reports receiving personal fees from EA Pharma Co., Ltd., AbbVie GK., Mitsubishi Tanabe Pharma Corporation, Janssen Pharmaceutical K.K., Mochida Pharmaceutical Co., Ltd., Takeda Pharmaceutical Co., Ltd. KH reports receiving research funding from EA Pharma Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Takeda Pharmaceutical Co., Ltd. HN reports receiving personal fees from AbbVie GK., Kissei Pharmaceutical Co., Ltd., Kyorin Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Corporation, Janssen Pharmaceutical K.K., Takeda Pharmaceutical Co., Ltd., Pfizer Lnc., Cell gene Corporation., EA Pharma Co., Ltd., Zeria Pharmaceutical CO., Ltd., Mochida Pharmaceutical Co., Ltd., Nippon Kayaku Co., Ltd., Daiichi Sankyo Company, Limited., JIMRO Co., Ltd., as well as grants for commissioned/joint research from Hoya Group Pentax Medical, Boehringer Ingelheim GmbH, Bristol-Myers Squibb Company. All other authors declare no competing interests. Ethics Approval Statement: The protocol of the clinical trial was approved by the IRB at each institution. Informed consent to participate in the study was obtained from each participant before inclusion.

Lascufloxacin hydrochloride to treat bacterial infection.

Lascufloxacin hydrochloride (AM-1977) is a novel 8-methoxy fluoroquinolone antibacterial agent with a unique pharmacophore at the 1st and 7th positions of the quinoline nucleus developed by Kyorin Pharmaceutical Co., Ltd. (Tokyo, Japan). It has been approved by the Japanese Ministry of Health, Labour and Welfare (MHLW) for treatment of respiratory tract and ear, nose and throat infections including community-acquired pneumonia and otorhinolaryngological infections, and shows great promise against fluoroquinolone-resistant strains of major pathogens which infect the respiratory tract. It is suitable for treating infections caused by Staphylococcus, Streptococcus, Pneumococcus, Moraxella (Branhamella) catarrhalis, Klebsiella, Enterobacter, Haemophilus influenzae, Legionella pneumophila, Prevotella and Mycoplasma pneumoniae that are sensitive to this drug.

441O - A platform trial with a registry study for rare cancers: MASTER KEY project

BackgroundRare cancers have had a challenge in establishing standard therapies for patients compared to major cancers, due to the lack of basis for clinical studies and investigations. We started a biomarker driven basket/umbrella trial using a “master protocol”, called the MASTER KEY Project, which aims to find more efficient ways to evaluate treatments for rare cancers. MethodsThe project opened in April 2017 and consists of a prospective registry study part and a multiple clinical trials part. Patients with advanced rare cancers (annual incidence <6 / 100,000 population)/cancers of unknown primary/rare tissue subtypes of common cancers, undergoing a molecular diagnostic testing are enrolled. The primary objective of the registry study is to collect consecutive data on biomarker, patient background, and prognosis to build a database highly reliable for use as historical control data in future clinical trials. Sub-studies are placed under a “master protocol”, and each sub-study will enroll patients with the appropriate biomarker, regardless of tumor type. ResultsAs of Feb. 2019, 515 patients were enrolled in the project, and 493 were available for their backgrounds. Most frequent cancer types were: soft tissue sarcomas (33.7%), neuroendocrine tumors (7.3%), tumors of CNS (5.9%), salivary gland tumors (4.5%), etc. 346 patients had next generation sequencing testing results. The most common alterations were TP53 (32.4%), KRAS (10.4%), PIK3CA (9.5%), MDM2 (5.2%), CDK42 (4.9%), CDKN2A (4.9%), and RB1 (4.9%). 278 patients already had 6 months follow-up, and 116 patients received 181 systemic chemotherapy regimens after enrollment. Of those regimens, 25 were biomarker-based therapies, and median progression free survival was 8.1 months compared to 4.8 months with other cytotoxic chemotherapy regimens (Hazard ratio 0.61; 95% confidential interval 0.32-1.07). ConclusionsMASTER KEY Project is one of the largest platform trials focused only on rare cancers. The project provided the chance for rare cancer patients to receive biomarker-based treatment, and it showed better prognosis. This is a continuous project aimed to accelerate treatment development for rare cancers, with clinical trials directed towards new drug approvals. Clinical trial identificationUMIN000027552. Legal entity responsible for the studyThe authors. FundingAstellas Pharma, Bristol-Myers Squibb, Chugai Pharmaceutical, Daiichi Sankyo, Eisai, Kyorin Pharmaceutical, Novartis Parma, Ono Pharmaceutical, Otsuka Pharmaceutical, Pfizer, Taiho Pharmaceutical and Takeda Pharmaceutical. DisclosureAll authors have declared no conflicts of interest.

Salvage Radiotherapy +/- Hormone Therapy versus Hormone Therapy Alone for Prostate-Specific Antigen Failure after Radical Prostatectomy: A Randomised, Multicentre, Open-Label, Phase 3 Trial (JCOG0401)

Background: No standard therapy has been established for patients with prostate-specific antigen (PSA) failure after radical prostatectomy (RP). This study was designed to determine whether radiotherapy ± hormone therapy is superior to hormone therapy alone in such patients. Methods: This multicentre, randomised, open-label, unblinded, phase 3 trial included 36 institutions. Patients with localized prostate cancer [T1-2N0M0) treated by RP, pT0/2/3 and pN0/x] whose PSA concentrations had decreased to <0•1 ng/mL after RP, then increased to 0•4-1•0 ng/mL, were randomised to Arm A [hormone therapy 80 mg bicalutamide (BCL) monotherapy followed by luteinizing hormone-releasing hormone agonist in case of BCL failure], or Arm B [64·8 Gy of salvage radiotherapy (SRT) followed by the same regimen as in Arm A in case of SRT failure]. The primary endpoint was time to treatment failure (TTF) of BCL in the intention-to-treat. This trial is registered with UMIN-CTR, 00026. Findings: From May 2004 to May 2011, 210 patients (105 in each arm) were registered, median follow-up being 5·5 years. The TTF of BCL was significantly longer in Arm B (hazard ratio 0aƒ»56, 90% CI (0·40-0·77); one-sided p=0·001). Thirty-two of 102 patients (31%) in Arm B did not have SRT treatment failure. TTF of protocol treatment was significantly longer in Arm B, however, clinical relapse-free and overall survival did not differ between the arms. The most frequent Grade 3-4 adverse event was erectile dysfunction (83 patients [80%] in Arm A vs. 76 [74%] in Arm B). Interpretation: Initial SRT followed by hormone therapy in case of treatment failure should be standard therapy for PSA failure after RP. Funding: Health Sciences Research Grants for Clinical Research for Evidence Based Medicine and Grants-in-Aid for Cancer Research (016) from the Ministry of Health, Labour and Welfare, Japan. Declaration of Interest: AY has received personal fees from Astellas Pharma Inc, AstraZeneca, Bayer, Janssen Pharmaceutical K.K., Sanofi and Takeda Pharmaceutical Company Ltd,. MW has received personal fees from Chugai Pharmaceutical Co., Ltd. and Johnson & Johnson K.K. MEDICAL COMPANY. TF has received personal fees from Bayer AG, Janssen Pharmaceutical K.K., Nihon Medi-Physics Co., Ltd., AstraZeneca, Astellas Pharma Inc., Takeda Pharmaceutical Company Limited and Daiichi Sankyo Company Limited. KH has received personal fees from Kyowa Hakko Kirin Co. Ltd., Chugai Pharmaceutical Co. Ltd. Takeda Pharmaceutical C. Ltd., Sanofi, Bristol-Myers Squibb, MSD K. K., Pfizer, Bayer, and Astellas Pharma Inc. TI has received personal fees from Astellas Pharma Inc, Janssen Pharmaceutical K.K., Astra Zeneca, Takeda Pharmaceutical Company Ltd., Bayer, Sanofi, MSD K.K., Bristol-Myers Squibb, Ono Pharmaceutical Co., Ltd. and Pfizer Pfizer Inc. KF has received personal fees from Chugai Pharmaceutical Co., Ltd., NIppon Kayaku Co., Ltd., Ferring Pharmaceuticals, Pfizer Inc., Bayer, ASKA Pharmaceutical Co., Ltd., Daiichi Sankyo Company Ltd., Novartis International AG, SBI Pharma, KYORIN Pharmaceutical Co.,Ltd., MSD K.K., Tsumura & Co., Taiho Pharmaceutical Co., Ltd., FUJIFILM Corporation, Ono Pharmaceutical Co., Ltd., Sanofi, Asahi Kasei, Janssen Pharmaceutical K.K., Kissei Pharmaceutical Co., Ltd.. SE has received personal fees from Astellas Pharma Inc., Takeda Pharmaceutical Company Ltd., Daiichi Sankyo Company Ltd., FUJIFILM Toyama Chemical Co., Ltd., Olympus Corporation, Asahi Kasei Pharma Corporation, Nihon Medi-Physics Co., Ltd. Nippon Shinyaku Co., Ltd., Pfizer Inc., Sanofi, Janssen Pharmaceutical K.K., Kissei Pharmaceutical Co., Ltd. TH has received personal fees from Takeda Pharmaceutical Company Ltd., AstraZeneca, Astellas Pharma Inc, Janssen Pharmaceutical K.K., Sanofi and Bayer. KK. KK had received personal fees from Astellas Pharma Inc, AstraZeneca, Asahi Kasei Pharma Corporation, AMCO Inc. and Nihon Medi-Physics Co., Ltd. OI has received personal fees from Johnson & Johnson K.K. MEDICAL COMPANY, Kissei Pharmaceutical Co., Ltd., KYORIN Pharmaceutical Co.,Ltd., Pfizer Inc., Taiho Pharmaceutical Co., Ltd., Sanofi, Daiichi Sankyo Company Ltd., Takeda Pharmaceutical Company Ltd., Ono Pharmaceutical Co., Ltd., Novartis Pharma, , Asahi Kasei Pharma Corporation, Hisamitsu Pharmaceutical CO., Inc. Hisamitsu Pharmaceutical Co.,Inc. and AstraZeneca. NS has received grants and personal fees from Astellas Pharma Inc., Ono Pharmaceutical Co., Ltd., personal fees from Janssen Pharmaceutical K.K., Takeda Pharmaceutical Company Ltd., Astra Zeneca, Astellas Pharma Inc, Bayer., Pfizer Inc., Novartis International AG, Chugai Pharmaceutical Co., Ltd., MSD K.K. MS has received personal fees from Janssen Pharmaceutical K.K., Astellas Pharma Inc, Takeda Pharmaceutical Company Ltd. and AstraZeneca. YY has received personal fees from Intuitive Surgical Inc., Bristol-Myers Squibb and Takeda Pharmaceutical Company Ltd. KN has received personal fees from AstraZeneca. HF has received personal fees from Taiho Pharmaceutical Co., Ltd. and Chugai Pharmaceutical Co., Ltd. KT has received personal fees from AstraZeneca. YK has received personal fees from AstraZeneca, Janssen Pharmaceutical K.K., Takeda Pharmaceutical Company Ltd., Astellas Pharma Inc, Nippon Kayaku Co., Ltd. SN has received personal fees from Takeda Pharmaceutical Company Ltd., AstraZeneca, Astellas Pharma Inc, Janssen Pharmaceutical K.K., BrightPath Biotherapeutics, Pfizer Inc., Novartis International AG, Chugai Pharmaceutical Co., Ltd., MSD K.K. and Ono Pharmaceutical Co., Ltd. Ethical Approval: The study protocol was approved by each Institutional Review Board.

Vibegron: First Global Approval

Vibegron is a selective beta 3 adrenergic receptor (β3AR) agonist that is being developed in Japan jointly by Kyorin Pharmaceutical Co., Ltd and Kissei Pharmaceutical Co., Ltd and in other regions worldwide (except in several other Asian countries) by Urovant Sciences for the treatment of overactive bladder (OAB). Based on results from Japanese phase III trials, vibegron received approval in Japan in September 2018 for this indication. This article summarizes the milestones in the development of vibegron leading to this first global approval for the treatment of OAB.

Evaluation of Rapid Onset of Action of ICS/LABA Combination Therapies on Respiratory Function in Asthma Patients: A Single-Center, Open-Label, Randomized, Crossover Trial.

IntroductionProducts based on inhaled corticosteroid (ICS)/long-acting β2 agonist (LABA) combinations may provide different clinical benefits. This study was conducted to compare the rapid effects of three such combination products: formoterol/fluticasone (FFC) aerosol (pMDI), formoterol/budesonide (FBC) dry powder inhaler (DPI), and vilanterol/fluticasone furoate (VFC) DPI.MethodsThe study design was a three-armed, randomized, crossover study. Patients included in the study had stable moderate asthma, defined as an Asthma Control Questionnaire (ACQ) score ≤ 0.75, and were undergoing step 2 or 3 asthma treatment as defined by JGL2015. Subjects were treated with fluticasone propionate inhaled via Diskus® during a 2-week washout period before randomization. At visit 2, subjects were randomly assigned in a 1:1:1 ratio to FFC, FBC, or VFC, and evaluated for changes in pulmonary function over time. At visits 3 and 4, the treatment was switched to another ICS/LABA combination in a crossover manner after a 1-week washout period. Spirometry was performed pre-dose and at 3, 10, and 30 min post-dose, and forced oscillation was implemented pre-dose and at 1, 7, 15, and 60 min post-dose.ResultsFifteen outpatients (63.3 ± 9.5 years, ACQ: 0.13 ± 0.19) completed the study. ∆FEV1 at 3 min did not significantly differ among the three groups. Significant increases in FEV1 and %FEV1 from baseline were observed in the FFC (p = 0.004, 0.003), FBC (p = 0.014, 0.011), and VFC (p = 0.032, 0.023) groups at 30 min. Improvements in respiratory resistance at 5–20 Hz from baseline at 60 min, resonant frequency, respiratory system reactance at 5 Hz, and low-frequency reactance area from baseline were observed at 1 min in the FFC group (p = 0.014, 0.002, 0.027, 0.018, respectively).ConclusionFFC administered using a pMDI showed favorable delivery to peripheral airways and significantly more rapid action promptly after inhalation as compared with other ICS/LABA preparations inhaled using a DPI, thus broadening the potential therapeutic options for asthma.Trial Registration NumberUMIN000029379.FundingKyorin Pharmaceutical Co., Ltd.Electronic supplementary materialThe online version of this article (10.1007/s41030-018-0062-x) contains supplementary material, which is available to authorized users.

Nonalcoholic Fatty Liver Disease Is Associated with Type 2 Diabetes Mellitus Only in Individuals with Visceral Obesity

Nonalcoholic fatty liver disease (NAFLD) is considered to be a risk factor for the onset of type 2 diabetes mellitus (T2D), but no studies have considered the role of visceral fat (VF), which is strongly linked to both NAFLD and T2D, as a potential confounder. Therefore, it is not fully understood how NAFLD and T2D are associated. We previously reported that the risk of T2D onset and NAFLD were associated via VF accumulation. In this study, we further investigated the association between liver fat accumulation or NAFLD and T2D comorbidity, focusing on the presence or absence of visceral obesity (VO). The subjects were 367 individuals (179 men, 48.8%; mean age, 56.3 years), who received health check-ups at our medical examination center, MedCity21. In all subjects, quantitative live fat accumulation was evaluated by controlled attenuation parameter (CAP) value using Fibroscan®, and VF area at umbilical level (VFA) by abdominal CT scan as an index of VF accumulation. Those who habitually drank &amp;gt; 30 g/day of alcohol and those with confirmed liver diseases were excluded. CAP value ≥ 236 dB/m was defined as NAFLD, and VFA &amp;gt; 100 cm2 as VO. The association of CAP values or existence of NAFLD with T2D comorbidity was examined in the subgroups with and without VO (n = 101 and n = 266, respectively) by multivariate logistic analysis, with other factors adjusted. NAFLD was found 33.8% of subjects in non-VO group and 75.2% in VO group, respectively. In non-VO group, neither CAP value nor NAFLD showed significant association with T2D. In contrast, in VO group, the odds of T2D comorbidity were higher by 12.7% for each 10 dB/m higher CAP value, and NAFLD raised the odds of T2D comorbidity 5.2 times compared to non-NAFLD. These findings suggest that liver fat accumulation is associated with T2D via interaction with VF accumulation. In conclusion, both liver fat accumulation and NAFLD are associated with T2D comorbidity only in individuals with VO. Disclosure S. Fukumoto: Speaker's Bureau; Self; Astellas Pharma US, Inc.. Research Support; Self; Chugai Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Corporation. Speaker's Bureau; Self; Novo Nordisk Inc., Sanofi K.K.. Research Support; Self; Takeda Pharmaceuticals U.S.A., Inc., Taisho Toyama Pharmaceutical Co., Ltd. M. Emoto: Speaker's Bureau; Self; Eli Lilly and Company, Boehringer Ingelheim Pharmaceuticals, Inc., Sumitomo Dainippon Pharma Co., Ltd.. Research Support; Self; Astellas Pharma Inc, Ono Pharmaceutical Co., Ltd., Kissei Pharmaceutical Co., Ltd., Kowa Pharmaceutical Company Ltd., Taisho Toyama Pharmaceutical Co., Ltd., Mitsubishi Tanabe Pharma Corporation. K. Motoyama: None. T. Morioka: Consultant; Self; Ono Pharmaceutical Co., Ltd.. Research Support; Self; Novartis Pharma K.K., MSD K.K., Eli Lilly and Company, Sanofi K.K., Mitsubishi Tanabe Pharma Corporation. Speaker's Bureau; Self; Takeda Pharmaceuticals U.S.A., Inc., Novo Nordisk Inc., Kowa Pharmaceuticals America, Inc., Astellas Pharma US, Inc., Boehringer Ingelheim Pharmaceuticals, Inc., Sumitomo Dainippon Pharma Co., Ltd. T. Shoji: Speaker's Bureau; Self; Astellas Pharma Inc, Bayer Yakuhin Ltd. Research Support; Self; Bayer Yakuhin Ltd. Speaker's Bureau; Self; Kowa Pharamaceutical Co Ltd, Daiichi Sankyo Company, Limited, MSD K.K., Chugai Pharmaceutical Co., Ltd.. Research Support; Self; Chugai Pharmaceutical Co., Ltd.. Speaker's Bureau; Self; Kyowa Hakko Kirin Co., Ltd., Kissei Pharmaceutical Co., Ltd. K. Hirata: Speaker's Bureau; Self; AstraZeneca, Astellas Pharma Inc., Eisai Co., Ltd., KYORIN Pharmaceutical Co.,Ltd., Daiichi Sankyo Company, Limited, Sumitomo Dainippon Pharma Co., Ltd., TAIHO PHARMACEUTICAL Co.,Ltd., Chugai Pharmaceutical Co., Ltd., TEIJIN HOME HEALTHCARE LIMITED, Eli Lilly Japan K.K, Nippon Shinyaku Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., Pfizer Japan Inc.. Research Support; Self; KYORIN Pharmaceutical Co.,Ltd., Daiichi Sankyo Company, Limited, Chugai Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., MSD K.K.. Other Relationship; Self; AstraZeneca, GlaxoSmithKline K.K., Chugai Pharmaceutical Co., Ltd., Nippon Boehringer Ingelheim Co. Ltd., Novartis Pharma K.K., Santen Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Eli Lilly Japan K.K. M. Inaba: None.

Aerosol Deposition of Inhaled Corticosteroids/Long-Acting β2-Agonists in the Peripheral Airways of Patients with Asthma Using Functional Respiratory Imaging, a Novel Imaging Technology

Drug deposition in the peripheral airways has been reported to differ between pressurized metered dose inhalers (pMDIs) and dry powder inhalers (DPIs). This study was conducted to determine the drug deposition fraction from three inhaled corticosteroids/long-acting β2-agonists (ICS/LABA) combination products for inhalation in the peripheral airways. An airway deposition model was constructed using computed tomography (CT) images obtained in patients with asthma. Functional respiratory imaging was used to determine the airway deposition of Flutiform® (Kyorin Pharmaceutical), Symbicort® (AstraZeneca and Astellas Pharma), and Relvar® (GlaxoSmithKline K.K.) without actual drug inhalation based on airway images from CT and computational fluid dynamics using drug particle size. The mean drug deposition fractions and standard deviations (SDs) in the peripheral airways for Flutiform, Symbicort, and Relvar were 30.33% ± 1.89%, 18.95% ± 2.03%, and 6.58% ± 0.98% for ICS, and 29.17% ± 1.81%, 17.24% ± 1.87%, and 11.30% ± 1.42% for LABA, respectively, showing that significantly higher proportions of ICS and LABA were deposited to the peripheral airways from Flutiform than from Symbicort and Relvar (p < 0.001). The mean drug deposition fractions and SDs in the entire lung for Flutiform, Symbicort, and Relvar were 46.18% ± 1.29%, 28.85% ± 0.78%, and 13.01% ± 0.66% for ICS, and 44.34% ± 1.23%, 26.32% ± 0.71%, and 20.02% ± 0.80% for LABA, respectively, showing that significantly higher proportions of ICS and LABA were deposited to the lung from Flutiform than from Symbicort and Relvar (p < 0.001). Flutiform (pMDI) was shown to provide significantly higher drug deposition fraction to the central/peripheral airways than Symbicort (DPI) and Relvar (DPI). It is suggested that Flutiform may provide a potent therapeutic option for various asthma patients, including those who have difficulty using DPIs. UMI22840. Kyorin Pharmaceutical Co., Ltd.

Inhaler Operability and Patient Satisfaction Regarding Genuair® and Respimat® Inhalers for Chronic Obstructive Pulmonary Disease: A Randomized Crossover Study

Tiotropium via the Respimat® device, once daily, is a common treatment for patients with chronic obstructive pulmonary disease (COPD). Aclidinium via the Genuair® device, twice daily, is an alternative option. However, there are few studies comparing the two devices. We evaluated inhaler operability, patient satisfaction, and patient preference between Genuair and Respimat inhalers after 2 weeks of daily use. COPD patients were randomly assigned to a Genuair or Respimat treatment group at Week 0, and were switched to the other treatment at the end of Week 2. The primary endpoint was the correct operation rate of the devices. The secondary endpoints included number of instructions needed to correctly operate the devices, number of technique errors, overall patient satisfaction, and willingness to continue using a device. Fifty-four outpatients (mean age 74.3 ± 10.1 years, 52 men) were enrolled, and all completed the study. The correct operation rate for the Genuair device was significantly higher than for the Respimat device (96.0 ± 7.4% and 89.1 ± 12.8%, respectively; p < 0.001). Patients required significantly less instruction to handle the Genuair device correctly and made fewer technique errors compared to the Respimat device. The mean overall satisfaction scores were significantly better for Genuair than Respimat, and significantly more patients were willing to continue using the Genuair device. Our findings suggest that the Genuair device is suitable for COPD patients, especially for those who are less adherent to treatment because of difficulties in inhaler handling, compared with the Respimat device. Clinical Trials Registry (22399, UMI19373, 23/October/2015). Kyorin Pharmaceutical Co., Ltd.

Changes in the Function and Expression of T-Type and N-Type Calcium Channels in the Rat Bladder after Bladder Outlet Obstruction

We evaluated possible changes in the function and expression of T-type and N-type Ca(2+) channels in the bladder of rats with bladder outlet obstruction. Female Sprague Dawley® rats were divided into a group with bladder outlet obstruction created by partial urethral ligation and a sham operated group. Six weeks postoperatively we determined the mRNA expression of T-type and N-type Ca(2+) channels in the bladder, dorsal root ganglion and spinal cord. We also cystometrically investigated expression by intravenous administration of the T-Ca blocker RQ-00311610 or the N-type Ca(2+) channel blocker ω-conotoxin GVIA. We then performed in vitro functional studies of detrusor strips using these blockers. mRNA expression of T-type Ca(2+) channels in the bladder detrusor and mucosa layers, and the spinal cord dorsal horn, and N-type Ca(2+) channels in the whole bladder and detrusor layer, and the spinal cord dorsal horn was greater in the obstructed group than the sham operated group. In obstructed rats bladder capacity and voided volume increased after RQ-00311610 administration but the number of nonvoiding contractions decreased after ω-conotoxin GVIA administration. Detrusor strips from obstructed rats showed weaker contractile responses to electrical field stimulation, particularly in regard to the purinergic component. ω-Conotoxin GVIA suppressed electrical field stimulation induced contractions only in the detrusor of obstructed rats, especially the cholinergic component. Blocking T-type Ca(2+) channels increased bladder capacity while N-type Ca(2+) channel blockade inhibited nonvoiding contractions in rats with bladder outlet obstruction. Decreased bladder efferent neurotransmission occurred after bladder outlet obstruction, predominantly in its purinergic component and detrusor contractions via cholinergic neurotransmission were activated in a compensatory manner, probably via N-type Ca(2+) channel up-regulation.

Antimuscarinics Suppress Adenosine Triphosphate and Prostaglandin E2 Release From Urothelium With Potential Improvement in Detrusor Overactivity in Rats With Cerebral Infarction

Antimuscarinics improve detrusor overactivity. We evaluated the effects and action mechanisms of imidafenacin (Kyorin Pharmaceutical, Tokyo, Japan), a novel therapeutic agent for overactive bladder with antimuscarinic activity, on mediator release from urothelium and detrusor overactivity induced by cerebral infarction. Bladder hydrodistention was achieved by intravesical infusion of Krebs solution. Bladder adenosine triphosphate and prostaglandin E(2) were measured in the presence and absence of anticholinergics using luciferin-luciferase assay and enzyme-linked immunoassay, respectively. Cerebral infarction was induced in rats by occluding the left middle cerebral artery. The effects of intravenous imidafenacin on bladder function were examined using cystometry in rats with cerebral infarction and in those pretreated with resiniferatoxin. Increased intravesical adenosine triphosphate and prostaglandin E(2) were shown by induced distention of isolated rat bladders. Imidafenacin and darifenacin (Kemprotec, Middlesbrough, United Kingdom) significantly suppressed the increases in adenosine triphosphate and prostaglandin E(2). Decreased bladder capacity was observed in rats with cerebral infarction. Detrusor overactivity was suppressed with a minimum intravenous dose of 0.001 mg/kg imidafenacin. The effects of imidafenacin were not noted in rats pretreated with resiniferatoxin. Results support the hypothesis or suggest that imidafenacin improves cerebral infarction induced detrusor overactivity by suppressing peripheral C-fibers. This effect is thought to be associated with suppression of the release of adenosine triphosphate and prostaglandin E(2) from the urothelium.

Unsaturated FAs prevent palmitate-induced LOX-1 induction via inhibition of ER stress in macrophages

Palmitic acid (PA) upregulates oxidized LDL receptor-1 (LOX-1), a scavenger receptor responsible for uptake of oxidized LDL (oxLDL), and enhances oxLDL uptake in macrophages. However, the precise underlying mechanism remains to be elucidated. PA is known to induce endoplasmic reticulum (ER) stress in various cell types. Therefore, we investigated whether ER stress is involved in PA-induced LOX-1 upregulation. PA induced ER stress, as determined by phosphorylation of PERK, eIF2α, and JNK, as well as induction of CHOP in macrophage-like THP-1 cells. Inhibitors [4-phenylbutyric acid (PBA), sodium tauroursodeoxycholate (TUDCA), and salubrinal] and small interfering RNA (siRNA) for the ER stress response decreased PA-induced LOX-1 upregulation. Thapsigargin, an ER stress inducer, upregulated LOX-1, which was decreased by PBA and TUDCA. We next examined whether unsaturated FAs could counteract the effect of PA. Both oleic acid (OA) and linoleic acid (LA) suppressed PA-induced LOX-1. Activation of the ER stress response observed in the PA-treated cells was markedly attenuated when the cells were cotreated with OA or LA. In addition, OA and LA suppressed thapsigargin-induced LOX-1 upregulation with reduced activation of ER stress markers. Our results indicate that activation of ER stress is involved in PA-induced LOX-1 upregulation in macrophages, and that OA and LA inhibit LOX-1 induction through suppression of ER stress.

Activity of Garenoxacin against Macrolide-Susceptible and -Resistant Mycoplasma pneumoniae

In the treatment of Mycoplasma pneumoniae infection, erythromycin and clarithromycin, 14-membered macrolides, and azithromycin, a 15-membered macrolide, are usually considered to be the first-choice drugs (4, 11). Macrolide antibiotics inhibit protein synthesis by binding to domain II and/or domain V of 23S rRNA (1, 13). We found that ca. 20% of M. pneumoniae strains isolated from patients in Japan from 2000 to 2003 were macrolide resistant (6, 9). Lucier et al. (5) and Okazaki et al. (7) found that an A-to-G transition or an A-to-C transversion at position 2063 (corresponding to position 2058 in Escherichia coli) or 2064 of the 23S rRNA gene results in a high level of resistance to macrolide antibiotics. Given this background, it was considered that an alternative to macrolides in the treatment of pneumonia caused by M. pneumoniae is needed. Garenoxacin, a des-F(6)-quinolone that exhibits excellent activity against respiratory pathogens such as Streptococcus pneumoniae and Chlamydophila pneumoniae, is under development for both oral and parenteral administration (10). In the present study, we examined the in vitro activity of garenoxacin against M. pneumoniae isolates, including macrolide-resistant strains, and compared it with those of other antibiotics. The following agents were employed for MIC determinations: garenoxacin (Toyama Chemical Co., Ltd., Tokyo, Japan), gatifloxacin (Kyorin Pharmaceutical Co., Ltd., Tokyo, Japan), levofloxacin and clarithromycin (LKT Laboratories, Inc., Saint Paul, MN), and minocycline (Lederle-Japan Ltd., Tokyo, Japan). The purity of each of these agents was above 99.8%. The following isolates of M. pneumoniae were tested: three macrolide-susceptible reference strains, FH, Mac, and M129, and four macrolide-resistant clinical isolates, 1 and 6 (isolates carrying the A-to-G mutation at position 2063 in the 23S rRNA gene for which the MIC of clarithromycin is 32 μg/ml) and 2 and 4 (isolates carrying the A-to-G mutation at position 2064 in the 23S rRNA gene for which the MIC of clarithromycin is 8 μg/ml). The PCR amplification and sequencing of domain II of the 23S rRNA genes were performed by a method reported previously (6). A broth microdilution method was used to determine the MICs. Serial twofold dilutions of antibacterial agents prepared in PPLO broth (Difco Inc., Detroit, MI) containing 104 to 105 CFU of M. pneumoniae/ml were put into 96-well microplates. The microplates were sealed with adhesive sheets and incubated at 37°C for 4 to 8 days. The MIC was defined as the lowest concentration of a drug at which the metabolism of the organism was inhibited, as evidenced by a lack of color change in the medium at the time when the drug-free control first showed a color change. The MICs of garenoxacin for the M. pneumoniae isolates, including the macrolide-resistant isolates, were 0.016 to 0.031 μg/ml, two- to fourfold lower than those of gatifloxacin and 8- to 32-fold lower than those of levofloxacin and minocycline (Table ​(Table1).1). The results for macrolide-susceptible strains such as M. pneumoniae FH, a reference strain, accorded with those given in previous reports (2, 8, 12). TABLE 1. Activities of various antibiotics against M. pneumoniae strains The newer fluoroquinolones such as gatifloxacin and moxifloxacin exhibit potent in vitro activity against a broad spectrum of organisms, including M. pneumoniae (3). Garenoxacin showed more-potent in vitro activity against M. pneumoniae than gatifloxacin and has attracted interest as a potential therapy for community-acquired pneumonia. These data indicate that garenoxacin may be a viable alternative to macrolides, such as clarithromycin, in the treatment of pneumonia caused by M. pneumoniae, including macrolide-resistant strains.