Schering-Plough Research Institute Research Articles

Interview: Drug Discovery and Development in the Field of Cardiovascular Disease

William Greenlee, the founder and president of MedChem Discovery Consulting LLC, has over three decades of research experience in industry at Merck Research Laboratories and the Schering-Plough Research Institute. He has contributed to the advancement of 20 drug candidates into preclinical development, 11 of which have progressed into clinical studies. Dr Greenlee was a member of the team that discovered the marketed drugs Vasotec™ and Prinivil™, and has led research teams that discovered several drug candidates that are now in clinical trials. His achievements and significant contributions to the field have been recognized with the presentation of a number of local and national awards, including the Alfred Burger Award in Medicinal Chemistry from the American Chemical Society and the Pennsylvania Drug Discovery Institute Drug Discovery Award. He was inducted into the American Chemical Society Medicinal Chemistry Hall of Fame in 2006. Dr Greenlee was elected a Fellow of the American Association for the Advancement of Science in 2007 and an American Chemical Society Fellow in 2009.

Pre-treatment with flucloxacillin does not affect the sugammadex-induced recovery from vecuronium-induced neuromuscular blockade in the rat

Department of Pharmacology, Schering-Plough Research Institute, a Division of Merck Sharp and Dohme, Newhouse, United Kingdom

Phase I/II study of the combination of decitabine (DAC) and temozolomide (TMZ) in patients (pts) with metastatic melanoma (MM).

8533 Background: TMZ has clinical activity at least equal to that of dacarbazine in MM. The DNA repair protein, O6- methylguanine-DNA-methyltransferase (MGMT), is implicated in resistance to TMZ. A functional DNA mismatch repair (MMR) pathway is required to initiate TMZ-induced cell death; MMR deficiency induced by promoter hypermethylation is a known mechanism of resistance. We hypothesized that combining of TMZ with DAC will induce dual modulation of DNA repair by depleting MGMT and re-expressing MMR. We now report results of a phase I/II study of TMZ and DAC in MM. Methods: In the phase I portion, pts were treated on 2 DAC dose levels (DL): 0.075 mg/kg IV qd and 0.15 mg/kg IV qd. DAC was administered qd × 5 d/wk for 2 wks, and TMZ was administered PO at 75 mg/m2 qd for 4 wks, starting on wk 2 of a 6-wk cycle. Phase II used a Simon 2-stage design: 14 eligible pts were to be accrued, and if ≥ 1 objective response (OR) was observed, the study would accrue an additional 20 pts. PK sampling was performed on Days 1 and 8; PBMC and tumor tissues were obtained on Days 1, 8, 15 and 29 of cycle 1. Results: 4 pts (3 evaluable) were accrued to DL1, and no DLTs were observed. There were no DLTs in the 6 pts accrued to DL2, which was declared the recommended Phase II dose (RP2D). 13 pts (12 evaluable) have been accrued to the Phase II portion, 1 PR has been observed, and the trial has proceeded to full accrual. Of the 18 pts treated at the RP2D, 9 (50%) have had stable disease for > 12 wks and continue to be on treatment. 3 (17%) have had SD > 24 wks. Grade ¾ neutropenia has been observed in 61% of pts but was reversible and in only 1 pt lasted > 7 d. 2 episodes of febrile neutropenia have been observed in the 18 pts at the RP2D. Serial tumor biopsies obtained from 2 pts confirmed the hypomethylating effect of DAC in melanoma. Conclusions: The combination of DAC and TMZ can be administered safely at biologically relevant dosages. The dual DNA repair modulation approach to the abrogation of chemotherapy resistance appears promising. Accrual is ongoing and will be completed by March 2010. Support: Eisai, Inc., Schering-Plough Research Institute, and University of Pittsburgh Cancer Institute, Melanoma and Skin Cancer Program P50 CA121973. Author Disclosure Employment or Leadership Position Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Eisai, Schering-Plough Research Institute

Potential synergistic activity of antimycotic substances in combination with human platelets against Aspergillus fumigatus

Sir, Invasive aspergillosis is increasingly recognized in immunocompromised hosts. Patients with prolonged and deep granulocytopenia following chemotherapy for haemato-oncological disorders or allogeneic bone marrow transplant recipients are particularly at risk. Platelets are second in abundance to red blood cells in the circulation, with 1.5–4.0×10 platelets/mL of blood in healthy individuals. They are essential components in haemostasis, but they also play an important role in antimicrobial host defence. Recently, we observed that platelets released serotonin after contact with aspergilli. Furthermore, platelets attenuated the virulence of Aspergillus spp. as the cell wall compound galactomannan was significantly decreased. Several in vitro studies have indicated that immune effector cells in combination with antifungals exert synergistic effects on aspergilli. However, no data are available on the effects of platelets and antimycotics on aspergilli. Two clinical Aspergillus fumigatus isolates and four antifungal agents were studied: amphotericin B (Sigma–Aldrich, Vienna, Austria); voriconazole (kindly provided by Pfizer, Vienna, Austria); posaconazole (kindly provided by Schering-Plough Research Institute, Kenilworth, NJ, USA); and caspofungin (MSD, Vienna, Austria). The MICs of all drugs used in this study were 0.5 mg/L for both isolates. Based on the MICs, drug concentrations of 0.5 and 1 mg/L were used for the platelet/drug studies. Fresh platelets were provided from the local Department of Immunology and Blood Transfusion. Platelets were collected from healthy donors by thrombocytopheresis using an Amicus cell separator (Baxter, Vienna, Austria). One hundred microlitres each of platelets (1×10/mL) and conidia (1×10/mL) in an effector/target ratio of 100:1 and specific concentrations of antifungals (0.5 and 1 mg/L) were inoculated into microwell plates (Greiner, Vienna, Austria) and incubated at 378C. The morphology of the A. fumigatus strains treated either with the combination of platelets plus antifungals or with each substance or platelets alone was investigated by assessing the germination rate and hyphal elongation, as described previously. Hyphal damage was assessed by a colorimetric assay with the dye 2,3-bis[2-methoxy-4-nitro-5-sulphophenyl]-2-H-tetrazolium-5carboxynilide sodium salt (XTT; Sigma–Aldrich) plus 40 mg/mL coenzyme Q (Sigma–Aldrich) and antifungal activity was calculated as the percentage of hyphal damage. Each experiment was performed with platelets from one donor using duplicate or quadruplicate wells for each condition and strain, and repeated six times. Differences between mean values of both strains were statistically evaluated by repeated-measures analysis of variance followed by Dunnett’s correction for multiple comparisons. A twosided P value of ,0.05 indicated statistical significance. Human platelets plus amphotericin B achieved significantly greater inhibition of the germination rate (P,0.05) than did amphotericin B or platelets alone (Figure 1a). The effect was additive. These results were found with both tested amphotericin B concentrations, although 1 mg/L revealed a better inhibitory effect than the lower concentration (Figure 1a). With caspofungin, the germination rate was not significantly reduced (Figure 1b). When azoles were used in combination with platelets, the inhibitory effect was found to be additive in comparison with either azole or platelets alone (Figure 1c and d). As found for the germination rate, human platelets plus amphotericin B achieved significantly greater inhibition of hyphal elongation (P,0.05) than amphotericin B or platelets alone. Hyphal elongation was significantly reduced in all tested aspergilli, either under platelet or caspofungin treatment. When used in combination, the effect was found to be additive. The combination of platelets plus either azole at any concentration tested did not significantly enhance the reduction of hyphal elongation. Platelets decreased the ability of hyphae to reduce XTT; however, the combination of platelets plus antimycotics had no additive effect on hyphal damage (data not shown). Our findings indicate that platelets in combination with antimycotics exert additive effects in reducing the germination rate and hyphal elongation of A. fumigatus in vitro. Among the tested antimycotic substances, amphotericin B revealed the best results in combination with human platelets. However, platelets plus antimycotics were not additive for hyphal damage. In the immunocompromised patient, inhaled Aspergillus conidia germinate into hyphae, the growing and invading structures of filamentous fungi. Consequently, blocking fungal germination and delaying hyphal growth is crucial in preventing invasive disease. In our study, the combination of platelets plus amphotericin B synergistically enhanced the antifungal activity (P,0.05) in reducing germination rate and hyphal elongation. Amphotericin B is known to complex with sterols in the fungal cell membrane, leading to pore formation and

SCH529074, a Small Molecule Activator of Mutant p53, Which Binds p53 DNA Binding Domain (DBD), Restores Growth-suppressive Function to Mutant p53 and Interrupts HDM2-mediated Ubiquitination of Wild Type p53

Abrogation of p53 function occurs in almost all human cancers, with more than 50% of cancers harboring inactivating mutations in p53 itself. Mutation of p53 is indicative of highly aggressive cancers and poor prognosis. The vast majority of mutations in p53 occur in its core DNA binding domain (DBD) and result in inactivation of p53 by reducing its thermodynamic stability at physiological temperature. Here, we report a small molecule, SCH529074, that binds specifically to the p53 DBD in a saturable manner with an affinity of 1-2 microm. Binding restores wild type function to many oncogenic mutant forms of p53. This small molecule reactivates mutant p53 by acting as a chaperone, in a manner similar to that previously reported for the peptide CDB3. Binding of SCH529074 to the p53 DBD is specifically displaced by an oligonucleotide with a sequence derived from the p53-response element. In addition to reactivating mutant p53, SCH529074 binding inhibits ubiquitination of p53 by HDM2. We have also developed a novel variant of p53 by changing a single amino acid in the core domain of p53 (N268R), which abolishes binding of SCH529074. This amino acid change also inhibits HDM2-mediated ubiquitination of p53. Our novel findings indicate that through its interaction with p53 DBD, SCH529074 restores DNA binding activity to mutant p53 and inhibits HDM2-mediated ubiquitination.

Ongoing Pregnancy Rates with Daily Recombinant Follicle-Stimulating Hormone/Gonadotropin-Releasing Hormone Antagonist Protocol in a Large North American Cohort of In Vitro Fertilization/Intracytoplasmic Sperm Injection Patients by Day of Embryo Transfer

BACKGROUND: The reference arm of a recent, randomized, double-blind, clinical trial (Engage) included a large cohort of patients from North American (NA) sites. Patients randomized to this reference arm were treated with a standardized rFSH/GnRH antagonist protocol but could have a day 3 or day 5 ET with one or two embryos transferred.

Sustained Virologic Response to Hepatitis C Virus Therapy and Spontaneous Viral Clearance Found Linked to Genotype

Sustained Virologic Response to Hepatitis C Virus Therapy and Spontaneous Viral Clearance Found Linked to Genotype

Membrane topology of human NPC1L1, a key protein in enterohepatic cholesterol absorption

The Niemann-Pick C1 Like 1 (NPC1L1) is a predicted polytopic membrane protein that is critical for cholesterol absorption. NPC1L1 takes up free cholesterol into cells through vesicular endocytosis. Ezetimibe, a clinically used cholesterol absorption inhibitor, blocks the endocytosis of NPC1L1 thereby inhibiting cholesterol uptake. Human NPC1L1 is a 1,332-amino acid protein with a putative sterol-sensing domain (SSD) that shows sequence homo-logy to HMG-CoA reductase (HMGCR), Niemann-Pick C1 (NPC1), and SREBP cleavage-activating protein (SCAP). Here, we use protease protection and immunofluorescence in selectively permeabilized cells to study the topology of human NPC1L1. Our data indicate that NPC1L1 contains 13 transmembrane helices. The NH2-terminus of NPC1L1 is in the lumen while the COOH-terminus projects to the cytosol. human NPC1L1 contains seven small cytoplasmic loops--four small and three large luminal loops--one of which has been reported to bind ezetimibe. Ezetimibe-glucuronide, the major metabolite of ezetimibe in vivo, can block the internalization of NPC1L1 and cholesterol. The membrane topology of NPC1L1 is similar to that of NPC1, and the putative SSD of NPC1L1 is oriented in the same manner as those of HMGCR, NPC1, and SCAP. The defined topology of NPC1L1 provides necessary information for further dissecting the functions of the different domains of NPC1L1.

Ergosterol biosynthesis and drug development for Chagas disease

This article presents an overview of the currently available drugs nifurtimox (NFX) and benznidazole (BZN) used against Trypanosoma cruzi, the aetiological agent of Chagas disease; herein we discuss their limitations along with potential alternatives with a focus on ergosterol biosynthesis inhibitors (EBI). These compounds are currently the most advanced candidates for new anti-T. cruzi agents given that they block de novo production of 24-alkyl-sterols, which are essential for parasite survival and cannot be replaced by a host's own cholesterol. Among these compounds, new triazole derivatives that inhibit the parasite's C14alpha sterol demethylase are the most promising, as they have been shown to have curative activity in murine models of acute and chronic Chagas disease and are active against NFX and BZN-resistant T. cruzi strains; among this class of compounds, posaconazole (Schering-Plough Research Institute) and ravuconazole (Eisai Company) are poised for clinical trials in Chagas disease patients in the short term. Other T. cruzi-specific EBI, with in vitro and in vivo potency, include squalene synthase, lanosterol synthase and squalene epoxidase-inhibitors as well as compounds with dual mechanisms of action (ergosterol biosynthesis inhibition and free radical generation), but they are less advanced in their development process. The main putative advantages of EBI over currently available therapies include their higher potency and selectivity in both acute and chronic infections, activity against NFX and BZN-resistant T. cruzi strains, and much better tolerability and safety profiles. Limitations may include complexity and cost of manufacture of the new compounds. As for any new drug, such compounds will require extensive clinical testing before being introduced for clinical use, and the complexity of such studies, particularly in chronic patients, will be compounded by the current limitations in the verification of true parasitological cures for T. cruzi infections.

Golimumab in patients with active rheumatoid arthritis after treatment with tumour necrosis factor α inhibitors (GO-AFTER study): a multicentre, randomised, double-blind, placebo-controlled, phase III trial

Tumour necrosis factor alpha (TNFalpha) inhibitors are frequently used to treat rheumatoid arthritis, but whether use of a different TNFalpha inhibitor can improve patient response is unknown. We assess the efficacy and safety of the TNFalpha inhibitor golimumab in patients with active rheumatoid arthritis who had previously received one or more TNFalpha inhibitors. 461 patients with active rheumatoid arthritis from 82 sites in 10 countries were randomly allocated by interactive voice response system, stratified by study site and methotrexate use, to receive subcutaneous injections of placebo (n=155), 50 mg golimumab (n=153), or 100 mg golimumab (n=153) every 4 weeks between Feb 21, 2006, and Sept 26, 2007. Allocation was double-blind. Eligible patients had been treated with at least one dose of a TNFalpha inhibitor previously. Patients continued stable doses of methotrexate, sulfasalazine, hydroxychloroquine, oral corticosteroids, and non-steroidal anti-inflammatory drugs. The primary endpoint was achievement at week 14 of 20% or higher improvement in American College of Rheumatology criteria for assessment of rheumatoid arthritis (ACR20). At week 16, patients who had less than 20% improvement in tender and swollen joint counts were given rescue therapy and changed treatment from placebo to 50 mg golimumab, or from 50 mg to 100 mg golimumab. Drug efficacy was assessed by intention to treat and safety was assessed according to the study drug given. This study is registered with ClinicalTrials.gov, number NCT00299546. Patients had discontinued previous TNFalpha inhibitors because of lack of effectiveness (269 [58%] patients) or reasons unrelated to effectiveness (246 [53%] patients), such as intolerance and accessibility issues. Patients had active disease, which was indicated by a median of 14.0 (IQR 9.0-22.0) swollen and 26.0 (16.0-41.0) tender joints for the whole group. 28 (18%) patients on placebo, 54 (35%) patients on 50 mg golimumab (odds ratio 2.5 [95% CI 1.5-4.2], p=0.0006), and 58 (38%) patients on 100 mg golimumab (2.8 [1.6-4.7], p=0.0001) achieved ACR20 at week 14. Two patients were never treated, and 57 patients did not complete the study because of adverse events, unsatisfactory treatment effect, loss to follow-up, death, or other reasons. 155 patients on placebo, 153 on 50 mg golimumab, and 153 on 100 mg golimumab were assessed for drug efficacy. For weeks 1-16, serious adverse events were recorded in 11 (7%) patients on placebo, 8 (5%) on 50 mg golimumab, and 4 (3%) on 100 mg golimumab. For weeks 1-24, after some patients were given rescue therapy, serious adverse events were recorded in 15 (10%) patients on placebo, 14 (5%) on 50 mg golimumab, and 8 (4%) on 100 mg golimumab. Golimumab reduced the signs and symptoms of rheumatoid arthritis in patients with active disease who had previously received one or more TNFalpha inhibitors. Centocor Research and Development and Schering-Plough Research Institute.

Cancer-induced activation of CD11b+Gr-1+ cells predispose hosts to antibody-independent anaphylactoid-type reactions (36.5)

Abstract Cancer can induce expansion of CD11b+Gr-1+ myeloid cells, attributed to immune suppression. We noted tumor-activation of these cells further correlated with predisposition for anaphylactoid-type reactions in mice bearing 4T1 mammary carcinomas, which induces strong CD11b+Gr-1+ expansion. To test this hypothesis, 4T1-tumor bearing (4T1+) mice were challenged with recombinant adenoviral vector (rAd), previously shown to activate myeloid lineage cells. When challenged with a single intravenous dose of rAd, 4T1+ mice succumbed to acute anaphylactoid-type reactions. In contrast, minimal responses were seen in rAd treated mice implanted with the related cell line, 66cl4, which do not induce CD11b+Gr-1+ cells. Liposome depletion prior to rAd challenge protected 4T1+ mice, whereas, passive transfer of CD11b+Gr-1+ cells from 4T1+ mice was sufficient to convey susceptibility for anaphylactoid reactions in normal mice. Metabolic pathways were upregulated for nitric oxide and leukotrienes in the 4T1-induced CD11b+Gr-1+ cells. Pre-treating 4T1+ mice with inhibitors of NO-synthetase and leukotrienes inhibited rAd-induced anaphylaxis. Thus, malignant tumor growth can alter CD11b+Gr-1+ myeloid cells, rendering hosts susceptible to anaphylactoid-type reactions. Importantly, there was no requirement for pre-existing Ag-specific IgE or IgG. This reflects clinical reports in which cancer patients undergo treatment-induced anaphylactoid responses in absence of pre-exposure and may provide insight for modeling alternative pathways induced by direct activation of inflammation- or tumor-modified myeloid cells. * Funded by the Schering-Plough Research Institute

The 20th Aspen Cancer Conference: Mechanisms of Toxicity, Carcinogenesis, Cancer Prevention, and Cancer Therapy 2005

The 20th Aspen Cancer Conference on Mechanisms of Toxicity, Carcinogenesis, Cancer Prevention, and Cancer Therapy was held from July 24 to 28, 2005 in Aspen, Colorado. Recognizing the importance of a 20-year history of excellence, the 2005 Aspen Cancer Conference included an extended scientific program as well as several special presentations and events. The scientific program, developed by a Scientific Advisory Committee co-chaired by Benjamin F. Trump (AMC Cancer Center) and Curtis C. Harris (National Cancer Institute, NIH), featured an extra day of seminars by former Aspen Cancer Conference Keynote Speakers. The returning keynote speakers included J. Michael Bishop, Douglas Hanahan, Thea Tlsty, Tony Hunter, and Scott Lowe. In a special public Conference Session held at the Aspen Institute, former NIH director Harold Varmus lectured on Oncogenes and the Future of Cancer Therapy, Ms. Karen Allred presented a piano recital, and former Ambassador to the Netherlands K. Terry Dornbush described A Survivor’s Story, cancer from the perspective of a two-time prostate cancer survivor. Other special programming in acknowledgement of the 20th year of the Aspen Cancer Conference included a whole day devoted to drug discovery with presentations from Cecil Pickett (Schering-Plough Research Institute), James Doroshow (National Cancer Institute), and Carl Barrett (Novartis) a review of the history of the Aspen Cancer Conference in the context of the history of Aspen, Colorado presented by Donald King (National Library of Medicine), and an awards ceremony honoring sustained contribution to the Aspen Cancer Conference by Theodore Puck, Benjamin J. Trump, and Donald King. The goals of the Aspen Cancer Conference are: to understand the mechanisms of toxicity and carcinogenesis, to understand and minimize cancer risk, to assess the public-health impact of cancer risk, and to develop the scientific basis for improving the diagnosis, prevention, and treatment of toxic and neoplastic disease. The Aspen Cancer Conference fosters intense interaction, discussion, and collaboration among key stakeholders in cancer research. One of the strengths of the Conference is the sustained participation in and support of the Conference by leaders in cancer research from government, academic, and industry sectors. Because of its unusual qualities, the Aspen Cancer Conference provides an environment that is uniquely conducive to generating novel insights related to current issues in cancer prevention, intervention, and therapy. The 20th Aspen Cancer Conference included seven Conference Sessions, a Poster Session by Aspen Cancer Conference Fellows and numerous informal discussions. Session topics included Progress in Drug Development, Metastasis, Inflammation and Cancer, Stress and Cancer, Risk Assessment, Technology Innovations and Apoptosis. At the close of the 2005 Conference, the Scientific Advisory Committee met to select topics for the 21st Aspen Cancer Conference which will held from July 16 to 18, 2006. This meeting summary concisely describes the content of each of the Conference Sessions and the scientific portions of the Special Sessions.

Structural identification of trace level enol tautomer impurity by on‐line hydrogen/deuterium exchange HR‐LC/MS in a LTQ‐Orbitrap hybrid mass spectrometer

Journal of Mass SpectrometryVolume 42, Issue 7 p. 967-970 JMS letter Structural identification of trace level enol tautomer impurity by on-line hydrogen/deuterium exchange HR-LC/MS in a LTQ-Orbitrap hybrid mass spectrometer Dr Guodong Chen Guodong Chen, Corresponding Author Dr Guodong Chen Guodong Chen guodong.chen@spcorp.com Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASchering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07 033, USA.===Search for more papers by this authorAnatoliy Khusid, Anatoliy Khusid Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorIbrahim Daaro, Ibrahim Daaro Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorPatrick Irish, Patrick Irish Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorBirendra N. Pramanik, Birendra N. Pramanik Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this author Dr Guodong Chen Guodong Chen, Corresponding Author Dr Guodong Chen Guodong Chen guodong.chen@spcorp.com Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASchering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07 033, USA.===Search for more papers by this authorAnatoliy Khusid, Anatoliy Khusid Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorIbrahim Daaro, Ibrahim Daaro Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorPatrick Irish, Patrick Irish Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this authorBirendra N. Pramanik, Birendra N. Pramanik Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, NJ 07033, USASearch for more papers by this author First published: 30 May 2007 https://doi.org/10.1002/jms.1232Citations: 16Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume42, Issue7July 2007Pages 967-970 RelatedInformation

To Make Better Therapeutics, Companies Strive to Increase the Content of Results and Get them Faster.

To Make Better Therapeutics, Companies Strive to Increase the Content of Results and Get them Faster.

SCH 503034, a Novel Hepatitis C Virus Protease Inhibitor, Plus Pegylated Interferon α-2b for Genotype 1 Nonresponders

SCH 503034 is a novel and potent oral hepatitis C virus (HCV) protease inhibitor. In this phase Ib study, we assessed safety parameters and virologic response of combination of SCH 503034 plus pegylated (PEG) interferon (IFN) alpha-2b in patients with HCV genotype 1 infections who were previously nonresponders to PEG-IFN-alpha-2b +/- ribavirin therapy. This was a multicenter, open-label, 2-dose level, 3-way crossover, randomized (to crossover sequence) study carried out in 3 medical centers in Europe. Adult patients received SCH 503034 200 mg (n = 14) or 400 mg (n = 12) 3 times daily orally and PEG-IFN-alpha-2b 1.5 microg/kg subcutaneously once each week. Patients received SCH 503034 as monotherapy for 1 week, PEG-IFN-alpha-2b as monotherapy for 2 weeks, and combination therapy for 2 weeks with washout periods between each treatment period. Combination therapy with SCH 503034 and PEG-IFN-alpha-2b was well tolerated, with no clinically significant changes in safety parameters. Mean maximum log(10) changes in HCV RNA were -2.45 +/- 0.22 and -2.88 +/- 0.22 for PEG-IFN-alpha-2b plus 200 mg and 400 mg SCH 503034, respectively, compared with -1.08 +/- 0.22 and -1.61 +/- 0.21 for SCH 503034 200 mg and 400 mg, respectively, and -1.08 +/- 0.22 and -1.26 +/- 0.20 for PEG-IFN-alpha-2b alone in the 200 mg and 400 mg SCH 503034 groups, respectively. SCH 503034 plus PEG-IFN-alpha-2b was well tolerated in patients with HCV genotype 1 nonresponders to PEG-IFN-alpha-2b +/- ribavirin. These preliminary results of antiviral activity of the combination suggest a potential new therapeutic option for this hard-to-treat, nonresponder patient population.

Mometasone Furoate Nasal Spray Rapidly and Enduringly Relieves Nasal Congestion and Other Symptoms of Seasonal Allergic Rhinitis

corticosteroids, was highly effective in the treatment of allergic rhinitis in children aged 6 to 12 years. It was also safe and well tolerated. Funding: Schering-Plough 251 Mometasone Furoate Nasal Spray Rapidly and Enduringly Relieves Nasal Congestion and Other Symptoms of Seasonal Allergic Rhinitis G. N. Gross, M. R. Danzig, D. Gates; Dallas Allergy and Asthma Center, Dallas, TX, Schering-Plough Research Institute, Kenilworth, NJ. RATIONALE: As one of the most bothersome symptoms of seasonal allergic rhinitis (SAR), congestion frequently interferes with sleep, daytime productivity/alertness, and quality of life. METHODS: This retrospective analysis of the onset and duration of efficacy of mometasone furoate nasal spray (MFNS) 200 mcg once daily over 15 days was based on pooled study data from 4 randomized, double-blind, placebo-controlled clinical trials in subjects with SAR. Subjects rated each symptom, including congestion, on a scale of 0 (none) to 3 (severe). Symptom severity ratings for the Total Nasal Symptom Score (TNSS) were categorized as mild (<6), moderate (6-<10), or severe ( 10). RESULTS: Mean baseline scores were similar for the MFNS (n5492) and placebo (n5497) groups for congestion (2.24 and 2.25, respectively) and TNSS (7.71 and 7.78, respectively). Treatment with MFNS produced significantly greater improvements than placebo in congestion and TNSS beginning on Day 2 and continuing on every day of the 15-day treatment period. At Day 2, score decreases with MFNS and placebo, respectively, were -0.36 vs -0.21 (-13.6% vs -7.7%; P<0.001) for congestion and -1.49 vs -0.81 (-16.5% vs -8.1%; P<0.001) for TNSS. At Day 15, decreases in scores with MFNS and placebo, respectively, were -0.82 vs -0.44 (-34.5% vs -17.2%; P<0.001) for congestion and -3.52 vs -1.94 (-41.6% vs -21.8%; P<0.001) for TNSS. CONCLUSIONS: MFNS rapidly improved congestion and total nasal symptoms in subjects with SAR, with significant improvements by Day 2 of treatment that continued for the duration of the study. Funding: Schering-Plough

Mometasone Furoate Nasal Spray Improves Nasal and Ocular Symptoms of Seasonal Allergic Rhinitis in Adolescent Patients

Symptoms of Seasonal Allergic Rhinitis in Adolescent Patients R. Anolik, D. Pearlman, M. Danzig, D. Gates; Allergy & Asthma Specialists, PC, Blue Bell, PA, Colorado Allergy and Asthma Centers, PC, Denver, CO, Schering-Plough Research Institute, Kenilworth, NJ. RATIONALE: In a population of adolescents, a demographic with a high incidence of seasonal allergic rhinitis (SAR), data for mometasone furoate nasal spray (MFNS) was examined. METHODS: Data from patients aged 12 to 17 years with symptomatic SAR enrolled in a phase III study were analyzed. These patients, who were part of a larger trial, were randomized to treatment with MFNS 200 mcg qd (n586) or placebo (n582) for 15 days. Symptom severity scores (05none; 35severe) were recorded in patient diaries twice daily. Endpoints included changes from baseline in total nasal symptom score (TNSS), as well as individual symptom scores and total ocular sign/symptom scores (TOSS). The endpoints were the same as those for the total study population. RESULTS: Over the 15 days of treatment, a significantly greater decrease from baseline in mean TNSS was observed with MFNS (-2.47; 28.8% decrease) compared with placebo (-0.9; 9.6% decrease) (P<0.001). In addition, MFNS demonstrated statistically significant improvements over placebo for each full day of the treatment period. Over the 15 days, improvements were seen with MFNS for each individual symptom: congestion (-0.66 vs -0.16; P<0.001); rhinorrhea (-0.63 vs -0.19; P<0.001); sneezing (-0.66 vs -0.26; P<0.001); and nasal itching (-0.52 vs -0.28; P50.03). The MFNS group demonstrated greater improvement in mean TOSS compared with the placebo group (-1.31 vs -0.55; P50.011). No serious adverse events were reported. CONCLUSIONS: In adolescents with symptomatic SAR, MFNS 200 mcg qd was shown to provide significant improvement in nasal and ocular J ALLERGY CLIN IMMUNOL VOLUME 119, NUMBER 1 Abstracts S61

Thrombin Receptor (Protease Activated Receptor-1) Antagonists as Potent Antithrombotic Agents with Strong Antiplatelet Effects

ADVERTISEMENT RETURN TO ISSUEPerspectiveNEXTThrombin Receptor (Protease Activated Receptor-1) Antagonists as Potent Antithrombotic Agents with Strong Antiplatelet EffectsSamuel ChackalamannilView Author Information Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, New Jersey 07033 Cite this: J. Med. Chem. 2006, 49, 18, 5389–5403Publication Date (Web):August 31, 2006Publication History Received29 March 2006Published online31 August 2006Published inissue 1 September 2006https://doi.org/10.1021/jm0603670Copyright © 2006 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views1765Altmetric-Citations72LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (298 KB) Get e-AlertsSUBJECTS:Aggregation,Antagonists,Cells,Peptides and proteins,Receptors Get e-Alerts

Recognition of Double-stranded RNA by Human Toll-like Receptor 3 and Downstream Receptor Signaling Requires Multimerization and an Acidic pH

Studies involving Toll-like receptor 3 (TLR3)-deficient mice suggest that this receptor binds double-stranded RNA. In the present study, we analyzed ligand/receptor interactions and receptor-proximal events leading to TLR3 activation. The mutagenesis approach showed that certain cysteine residues and glycosylation in TLR3 amino-terminal leucine-rich repeats were necessary for ligand-induced signaling. Furthermore, inactive mutants had a dominant negative effect, suggesting that the signaling module is a multimer. We constructed a chimeric molecule fusing the amino-terminal ectodomain of TLR3 to the transmembrane and carboxyl terminal domains of CD32a containing an immunoreceptor tyrosine-based motif. Expression of TLR3-CD32 in HEK293T cells and the myeloid cell line U937 resulted in surface localization of the receptor, whereas the nonrecombinant molecule was intracellularly localized. The synthetic double-stranded RNAs poly(I-C) and poly(A-U) induced calcium mobilization in a TLR3-CD32 stably transfected U937 clone but not in control cells transfected with other constructs. An anti-TLR3 antibody also induced Ca(2+) flux but only when cross-linked by a secondary anti-immunoglobulin antibody, confirming that multimerization by the ligand is a requirement for signaling. The inhibitors of lysosome maturation, bafilomycin and chloroquine, inhibited the poly(I-C)-induced biological response in immune cells, showing that TLR3 interacted with its ligand in acidic subcellular compartments. Furthermore, TLR3-CD32 activation with poly(I-C) was only observed within a narrow pH window (pH 5.7-6.7), whereas anti-TLR3-mediated Ca(2+) flux was pH-insensitive. The importance of an acidic pH for TLR3-ligand interaction becomes critical when using oligomeric poly(I-C) (15-40-mers). These observations demonstrate that engagement of TLR3 by poly(I-C) at an acidic pH, probably in early phagolysosomes or endosomes, induces receptor aggregation leading to signaling.

Food and Drug Administration Drug Approval Summary: Temozolomide Plus Radiation Therapy for the Treatment of Newly Diagnosed Glioblastoma Multiforme

On March 15, 2005, the U.S. Food and Drug Administration approved temozolomide (Temodar capsules, Schering-Plough Research Institute) for the treatment of adult patients with newly diagnosed glioblastoma multiforme concomitantly with radiotherapy and then as maintenance treatment. Five hundred seventy-three glioblastoma multiforme patients were randomized to receive either temozolomide + radiotherapy (n = 287) or radiotherapy alone (n = 286). Patients in the temozolomide + radiotherapy arm received concomitant temozolomide (75 mg/m2) once daily for the duration of radiation therapy (42-49 days). This was followed, 4 weeks later, by six cycles of temozolomide, 150 or 200 mg/m2 daily for 5 days, every 4 weeks. Patients in the control arm received radiotherapy only. In both arms, radiotherapy was delivered as 60 Gy/30 fractions to the tumor site with a 2 to 3 cm margin. Pneumocystis carinii pneumonia prophylaxis was required during temozolomide + radiotherapy treatment and was continued until recovery of lymphocytopenia (Common Toxicity Criteria grade <1). At disease progression, temozolomide salvage treatment was given to 161 of 282 patients (57%) in the radiotherapy alone arm, and to 62 of 277 patients (22%) in the temozolomide + radiotherapy arm. Patients receiving concomitant and maintenance temozolomide + radiotherapy had significantly improved overall survival. The hazard ratio was 0.63 (95% confidence interval, 0.52-0.75; log-rank, P < 0.0001). Median survival was 14.6 months (temozolomide + radiotherapy) versus 12.1 months (radiotherapy alone). Adverse events during temozolomide treatment included thrombocytopenia, nausea, vomiting, anorexia, constipation, alopecia, headache, fatigue, and convulsions.