Chiron Corporation Research Articles

Complete Remission after the First Cycle of Induction Chemotherapy Determines the Clinical Efficacy of Relapse-Preventive Immunotherapy in Acute Myeloid Leukemia

Background: Contemporary studies have identified >1 cycle of induction chemotherapy to induce complete remission (CR) as an independent risk factor for relapse and mortality in younger adults with acute myeloid leukemia (AML; Othus et al., Leukemia 2019). This study aimed at defining the potential impact of previous induction chemotherapy on the clinical benefit of relapse-preventive immunotherapy in AML. Methods: In a previously reported phase 3 trial (Brune et al., Blood 2006; Martner et al., Blood Rev 2013), two hundred and sixty-one patients with AML in first CR (18-84 years, median 55) who were not eligible for allogeneic transplantation were randomly assigned to receive immunotherapy with histamine dihydrochloride and low-dose interleukin-2 (HDC/IL-2) or standard of care (no treatment, control). The HDC component of this regimen aims at countering immunosuppression induced by reactive oxygen species, which are formed by the NOX2 enzyme expressed by normal and malignant myeloid cells. HDC, a NOX2 inhibitor, improves the function and viability of anti-leukemic lymphocytes such as natural killer cells and cytotoxic T cells (Martner et al., J Pathol 2019). The immunotherapy was initiated in CR1 after the completion of chemotherapy with the aim to prevent relapse in the post-consolidation phase. Cycles 1 to 3 comprised 3 weeks on treatment and 3 weeks off treatment. In cycles 4 to 10 the off-treatment periods were extended to 6 weeks. In each cycle, patients in the treatment arm received HDC (Noventia Pharma, Milan, Italy) at 0.5 mg and human recombinant low-dose IL-2 (aldesleukin; 16 400 IU/kg; Chiron Corporation, Emeryville, CA) subcutaneously twice a day. All patients were followed for at least 36 months after random assignment. The median follow-up time was 48 months. Results: The number of previous induction cycles was known in 260 patients, and 203 of these attained CR after 1 induction cycle (78%). Among the 57 remaining patients, forty-seven attained CR after 2 cycles, and 3 (n=7) or 4 (n=3) cycles were required to induce CR in 10 patients. We compared outcomes of patients who had achieved CR1 after 1 vs >1 cycle of induction chemotherapy and observed a non-significant trend towards inferior outcome in control arm patients who had required >1 induction with a similar trend in younger control patients (<60 years old). In the HDC/IL-2 arm, outcome was significantly superior in younger patients who had attained CR1 after 1 vs >1 induction (leukemia-free survival, LFS, P=0.0007, hazard ratio, HR, 1.67-6.77; overall survival, OS, P=0.009, HR 1.30-6.33 in multivariable analyses correcting for potential confounding factors for relapse and death). When comparing outcomes between treatment and control arms within groups of patients who had attained CR1 after 1 cycle of induction chemotherapy, we observed that the LFS of patients in the HDC/IL-2 arm was significantly superior over controls (P=0.01 in multivariable analysis, N=203). In patients who were <60 years old at random assignment (N=130), treatment with HDC/IL-2 entailed significantly improved LFS (P=0.0008 vs control, HR 0.29-0.72) and OS (P=0.01, HR 0.28-0.75). HDC/IL-2 did not improve LFS or OS in patients who had required >1 cycle of induction to attain CR1 (P>0.5 vs control) and was not significantly beneficial in older patients. A test of interaction supported that the effect size of HDC/IL-2 vs control for LFS was more pronounced in younger patients who had attained CR after 1 (HR=0.48) vs >1 (HR=1.33) cycle of induction (P=0.04). Figure 1 shows Kaplan-Meier analyses of outcome, with unadjusted P-values from logrank tests, in the phase 3 trial arms among all patients attaining CR1 after 1 induction (A and B), in corresponding patients <60 years old (C and D) and in all patients who had attained CR1 after >1 cycle of induction (E and F). Conclusions: These findings imply that the efficiency of previous induction chemotherapy independently determines the clinical benefit of relapse-preventive immunotherapy in adult AML patients who are not candidates for allogeneic transplantation. The results may have implications for the selection of patients who are likely to benefit from treatment with HDC/IL-2 and for the design of clinical trials that evaluate relapse-preventive immunotherapy. Figure 1 Disclosures Thorén: Cytovia, New York: Patents & Royalties: Holds patents protecting the use of NOX2 inhibitors in cancer. Martner:Cytovia, New York: Patents & Royalties: Holds patents protecting the use of NOX2 inhibitors in cancer. Hellstrand:Cytovia, New York: Patents & Royalties: Holds patents protecting the use of NOX2 inhibitors in cancer. OffLabel Disclosure: HDC/IL-2 is approved for the prevention of AML relapse within the EU but is not yet approved in the US.

Therapy of Chronic Hepatitis C in the Era of Nanotechnology: Drug Delivery Systems and Liver Targeting.

Since the British scientist Michael Houghton along with George Kuo, Qui-Lim Choo (Chiron Corporation Emeryville), and Daniel W. Bradley (Centers for Disease Control and Prevention) codiscovered the causative agent of hepatitis C in 1989, so much progress has been made for the screening of blood donors and management of this chronic liver disease. In this regard, direct-acting antiviral agents (DAAs) have emerged as the potential "cure" of this slowly progressing and devastating disease. However, improvements are still clearly required since the anti-hepatitis C drugs currently available in the market are so extremely expensive (i.e. $94,500 for a 12-week course of treatment), that many patients will have a denied access to such drugs by their insurers. </p> <p> In the last few years, nanotechnology has emerged as a new platform for drug development, contributing significantly to the improvement of the administration and delivery of many drugs. Additionally, nanotechnologies can provide unique solutions even in poorer societies. </p> <p> This manuscript reviews the current knowledges on the available anti-hepatitis C drugs and the new drug candidates being investigated as well, and introduces the recent advances in nanocarrier-based delivery systems. Finally, the challenges in the development of drug delivery systems for the targeting of antiviral drugs to the liver are also discussed.

An interview with Margaret A Liu: the future of gene-based vaccines and immunotherapies, and other musings

Interview by Jenaid Rees, commissioning editor.Margaret A Liu is best known for her pioneering work in the area of DNA vaccines. A world renowned scientist, Liu was named one of ‘The 50 Most Important Women Scientists’ by Discover magazine in 2002. Liu obtained her M.D. from Harvard Medical School, and has held positions at numerous institutions including Harvard Medical School, Massachusetts Institute of Technology, University of Pennsylvania, UCSF, and the Karolinska Institutet in Stockholm. In her career she has served as Senior Advisor in Vaccinology at the Bill & Melinda Gates Foundation and Executive Vice-Chair of the International Vaccine Institute in Seoul, Korea and worked for companies including Merck, Transgène and Chiron Corporation. Her research achievements have led to her receipt of honorary lectureships, and she has held many board positions throughout her career. Liu currently consults in the fields of vaccines and immunotherapy for companies, universities, and non-governmental and governmental scientific organizations, and is a Foreign Adjunct Professor at the Karolinska Institutet in Stockholm, and an Adjunct Professor at the University of California, San Francisco.

Professor Frank McCormick: K-Ras proteins offer new opportunities for therapeutic intervention against human cancers.

Frank McCormick, PhD, FRS, is Professor Emeritus of the UCSF Helen Diller Family Comprehensive Cancer Center (Figure 1). Prior to joining the UCSF faculty, Prof. McCormick pursued cancer-related work with several Bay Area biotechnology firms and held positions with Cetus Corporation (Director of Molecular Biology, 1981-1990; Vice President of Research, 1990-1991) and Chiron Corporation, where he was Vice President of Research from 1991 to 1992. In 1992 he founded Onyx Pharmaceuticals, a company dedicated to developing new cancer therapies, and served as its Chief Scientific Officer until 1996. At Onyx Pharmaceuticals, he initiated and led drug discovery efforts that led to the approval of Sorafenib in 2005 for treatment of renal cell cancer, and for liver cancer in 2007, and the approval of ONYX-015 in 2006 in China for treatment of nasopharyngeal cancer. Sorafenib is being tested in multiple indications worldwide. In addition, Prof. McCormick’s group led to the identification of a CDK4 kinase inhibitor. Prof. McCormick’s current research interests center on the fundamental differences between normal and cancer cells that can allow the discovery of novel therapeutic strategies.

Resumen de congreso: 20 años después del primer Simposium Internacional sobre el virus de la hepatitis C y otros virus relacionados

The hepatitis C virus (HCV) was discovered by the team of Michael Houghton at Chiron Corporation in 1989 and the first symposium on HCV and related viruses was held in Venice, Italy, shortly after, in 1992. This conference was organized to advance knowledge on what then was a mysterious virus responsible for most cases of «non-A, non-B» hepatitis. During the 20 years since the first conference, the scientific quality of presentations has steadily increased, together with the tremendous advances in basic and clinical research and epidemiology. What started as a small conference on a new virus, about which there were very few data, has today become a first-in-class congress: a meeting place for basic researchers, clinicians, epidemiologists, public health experts, and industry members to present the most important advances and their application to HCV treatment and control. The nineteenth HCV symposium was held in September 2012, once again in Venice.

Vaccine adjuvants: the future is bright

Interview by Jenaid Rees, Commissioning EditorRino Rappuoli is the Global Head of Vaccines Research at Novartis Vaccines & Diagnostics (Siena, Italy). Previously, he was Head of R&D (Sclavo, Italy), and then Head of Vaccine Research and Chief Scientific Officer at Chiron Corporation (CA, USA). Rappuoli earned his PhD in Biological Sciences at the University of Siena (Siena, Italy) and has served as visiting scientist at Rockefeller University (NY, USA) and Harvard Medical School (Boston, MA, USA). His team developed the first recombinant bacterial vaccine (against pertussis) and the first conjugate vaccine against meningococcus C and A. He also worked on the MF59 adjuvant used in a vaccine against pandemic influenza and an experimental vaccine against Helicobacter pylori. Rappuoli and his team pioneered the genomic approach to vaccine development termed reverse vaccinology and used it to develop a vaccine against group B meningococcus. Among his scientific contributions, he was cofounder of the field of cellular microbiology, a discipline that has merged cell biology and microbiology. He is a member of numerous international associations, including the US National Academy of Sciences, the American Society for Microbiology and the European Molecular Biology Organization and serves on the Expert Review of Vaccines Editorial Board. He is also a recipient of several prestigious awards, including the Paul Ehrlich and Ludwig Darmstaedter Prize.

West Nile Virus Lineage 2 from Blood Donor, Greece

To the Editor: West Nile virus (WNV) is a mosquito-borne flavivirus that primarily causes an asymptomatic or mild disease in humans; however, in <1% of infected persons, it causes neurologic disease. The virus has received increased attention since 2002 when it was established that WNV is transmissible by blood transfusion and organ transplantation (1). A major WNV outbreak occurred in 2010 in Greece; most cases occurred in the northern part of the country (2). Of the 197 WNV neuroinvasive cases reported, 33 were fatal (3). Many nonneuroinvasive cases were observed (4). A lineage 2 WNV (Nea Santa-Greece-2010 strain) was detected in Culex pipiens mosquitoes collected at 2 locations where WNV cases had been reported (5). Although this strain shows high genetic identity to a Hungarian WNV strain isolated from birds in 2004, it has the amino acid substitution H249P in nonstructural protein 3 (NS3) (6). This mutation has been associated with increased virulence in WNV lineage 1 strains (7). Clinical WNV disease in humans had not been previously documented in Greece, and surveillance of blood donors in 2006 and 2007 did not show any WNV-positive result (8). On August 11, 2010, shortly after confirmation of the outbreak of WNV infections in humans in Greece, an action plan for the protection of blood safety was initiated. All donors were asked to report any fever-like illness up to 15 days after donation. Individual donation nucleic acid testing (NAT) of all blood donors living in the WNV-affected areas was implemented on August 22, 2010. The first WNV-positive by NAT result was obtained from a sample donated on August 22. Testing was performed by using the automated Procleix TIGRIS System (Chiron Corporation, Emeryville, CA, USA). The WNV-positive blood donor was a 40-year-old immunocompetent woman, a resident of a village in northern Greece. The village is located between 2 lakes, and the area is one of Europe’s major wetlands. The 2 locations where the WNV-positive mosquitoes were collected are near each other (Figure A1). The woman was working in an open-air fish market and reported numerous mosquito bites. At the time of blood donation, she was asymptomatic; 2 days later she had myalgia, arthralgia, and severe retro-orbital pain, lasting 2–3 days each. A second blood sample taken on August 26 was also NAT positive. Serologic testing for WNV IgM and IgG was performed by ELISA (WNV IgM Capture DxSelect and WNV IgG DxSelect; Focus Diagnostics Inc., Cypress, CA, USA). An index ≥1.5 for IgM and ≥1.1 for IgG was considered positive. No antibodies were detected in the initial and second serum samples; however, a third sample taken on September 20 was positive for WNV IgM and IgG (indices 4.7 and 3.8, respectively). Nested reverse transcription PCRs with the initial blood sample gave positive results (9,10). WNV lineage 2 sequences were obtained and were identical to those of the Nea Santa-Greece-2010 strain (6). One milliliter of each 1:10 and 1:100 dilution of whole blood in minimum essential medium containing antimicrobial drugs and 2% fetal bovine serum was placed on Vero E6 cell monolayers in 25-cm2 cell culture flasks. The procedure was performed in a biosafety level 3 laboratory, in which WNV lineage 2 had never been handled. After the sample was incubated 1 hour at 37°C in a 5% CO2 incubator, 9 mL of fresh medium containing 4% fetal bovine serum was added. Cytopathic effects were observed in the flask inoculated with the 1:100 dilution on day 3 after infection. An aliquot of supernatant was used to infect fresh cell monolayers. WNV growth in the cell culture was demonstrated by reverse transcription PCR and immunofluorescence assay. Sequences of the cell culture isolate were identical to those of the directly detected virus. To check whether the isolate possessed the H249P substitution, a set of nested primers spanning the region 5140–5660 from the WNV genome was designed: NS3a-5′-GCTGGCTTCGAACCTGA-3′ and NS3b-5′-CAATCATCGTTCTTGC-3′ for the first round PCR and NS3c-5′-GCTGCTGAGATGTCTGA-3′ and NS3d-5′-TCATATCCAGTGTTCCA-3′ for the nested PCR. The H249P substitution was present. Sequences were submitted to GenBank (accession nos. {type:entrez-nucleotide,attrs:{text:JF917091,term_id:349806861}}JF917091, {type:entrez-nucleotide,attrs:{text:JF917092,term_id:349806863}}JF917092). Virus isolation from WNV patients is usually unsuccessful because viremia levels are low and last only a short time. WNV strains are usually isolated from immunocompromised patients, blood donors, IgM-negative immunocompetent patients who seroconverted, or autopsy brain samples. For the donor reported here, WNV was isolated 2 days before illness onset, when no antibodies were present. The WNV-positive blood donor was detected 1 day after the introduction of blood screening. The early diagnosis of the initial human WNV cases in Greece, which resulted in prompt implementation of NAT testing, had a substantial positive impact on the safety of the blood supply in the affected areas. The risk for virus transmission was reduced for blood recipients, in particular those who receive multiple transfusions and immunocompromised patients in need of transfusion.

J. Craig Venter—The Human Genome Project

American-born biochemist-geneticist J. Craig Venter is acknowledged, along with geneticist Francis Sellers Collins (1950-), as being a primary force behind the Human Genome Project. Venter, with private funding, and Collins, with public funding, independently mapped and sequenced human DNA. Their reports appeared in 2001 (one in Nature and the other in Science). A sheet of 5 stamps (Scott No. 561) issued by Palau in 1999 provides an opportunity to honor J. Craig Venter indirectly. The sheet was issued to commemorate advances in science and medicine in the 20th century. One of the stamps representing the Human Genome Project shows the overall structure of DNA. J. Craig Venter was born on October 14, 1946, in Salt Lake City, UT. He spent his youth in San Francisco, CA, where he attended high school. After high school, he joined the US Naval Medical Corps and served in Vietnam, where he provided medical care at an orphanage. After Venter returned to the United States, he enrolled as a premedical student at the University of California, San Diego. However, he soon decided to pursue a career in research. He received a BS degree in biochemistry in 1972 and a PhD degree in physiology and pharmacology in 1975, both from the University of California, San Diego. In 1976, Venter became an assistant professor of pharmacology and therapeutics at the State University of New York at Buffalo and also worked at the nearby Roswell Park Memorial Institute. His research focused primarily on the elucidation of the structure and function of adrenergic receptors and muscarinic cholinergic receptors. Venter was promoted to professor in the early 1980s. For his work on receptors, he received the Boehringer Ingelheim Muscarinic Receptor Research Award. In 1984, he became a section chief at the National Institute of Neurological Disorders and Stroke at the National Institutes of Health (NIH) in Bethesda, MD, where he worked until 1992. His work at the NIH concentrated on neurotransmitters, specifically on genes that encode for enzymes involved in the synthesis of neurotransmitters and receptors. In July 1992, Venter left the NIH and became founder and chairman of the board of The Institute for Genomic Research (TIGR), a not-for-profit genomics research institution in Rockville, MD. He served as its president until 1998. In 1998, Venter joined Applera Corporation and became president and chief scientific officer of the newly founded Celera Genomics. The goal of Celera Genomics was to become the definitive source of genomic and related medical and biological information. In January 2002, Venter stepped down as president of Celera Genomics but continued to chair its scientific advisory board. Currently, he is serving as president of J. Craig Venter Institute. Venter pioneered the use of automated gene sequencers, and in 1990, he developed “expressed sequence tags” (ESTs), a new strategy for the discovery and tagging of genes that revolutionized biological science. In this approach, complementary DNA is partially sequenced, or “tagged,” using an automated DNA sequencing machine. The resulting sequences (ESTs) are long enough for one to be distinguished from another. Venter has published more than 160 research articles and has received many awards, including being elected a fellow of several societies, such as the American Association for the Advancement of Science and the American Academy of Microbiology. In 1999, he received the Beckman Award and the Chiron Corporation Biotechnology Research Award. Efforts to bring Collins and Venter together to complete the mapping of the human genome began in late 1999. In March 2000, US President Bill Clinton (1946-) and British Prime Minister Tony Blair (1953-) made a joint declaration that all genome information should be free to the public. This announcement led to cooperation between Collins and Venter, and on June 26, 2000, Venter and Collins jointly announced that, after nearly a decade of work, both the public Human Genome Project headed by Collins and Celera Genomics headed by Venter had deciphered essentially all the genes in human DNA. The importance of the cooperation between the 2 geneticists and the genome sequencing effort is far-reaching and may result in the discovery of keys to the diagnosis and treatment of numerous diseases, from diabetes to heart disease to Alzheimer disease.

Allogeneic NK Cell Therapy After Autologous Stem Cell Transplant: Results of a Phase I Study

AbstractAbstract 4299Previous reports have shown that infusion of NK cells from a MHC mismatched donor can mediate an anti-leukemic effect in the recipient of an allogeneic hematopoietic stem cell transplant (HSCT). In this phase I study, we infused increasing numbers of allogeneic NK-cell enriched mononuclear cells (NK-MNC) from a MHC haplo-mismatched relative into patients who had recently undergone autologous stem cell transplant. We sought to determine whether infusion of mismatched, allogeneic NK-MNC cells was safe without concern for GvHD or graft rejection, and also whether cell collection, processing and patient treatment could feasibly be performed at different cities across the US.MNC were obtained by apheresis from healthy haploidentical relatives by one steady-state leukapheresis of 2–4 hours on day 1, and were sent by air courier to the PACT* cell processing facility (University of Minnesota) where immunomagnetic depletion of CD3 cells (Miltenyi CliniMACS) was performed. The CD3-depleted cells were then cultured in X-VIVO 15, without gentamicin and phenol red (Cambrex BioScience, Walkersville, Maryland), supplemented with 1000 U/mL IL-2 (Chiron Corporation, Emeryville, CA) and 10% human heat-inactivated AB serum (Valley Biomedical Products and Services, Inc., Winchester, VA) in VueLife™Teflon® (FEP) bags (American Fluoroseal Corporation, Gaithersburg, MD). The resulting NK-MNC products were then returned by air courier at approximately body temperature to Boston for infusion on day 3. Twelve patients (age range: 27–63) within 49–191 days (median 106 days) after autologous HSCT were treated at four different dose levels of NK-MNC: 105, 106, 107 and 2×107 NK-MNC/kg. No logistical transport issues between Boston and the processing facility in Minnesota occurred. Release criteria (< 5 × 105 CD3+ cells/kg, > 20% CD3-/CD56+ cells, viability >70%, Gram stain – no organisms) were met in all but one case.Side effects after infusion occurred only at the higher dose level of NK-MNC infusion: rigors (n=2) and muscle aches (n=1), responsive to meperidine. None of the patients required discontinuation of NK-MNC infusion. No GvHD or marrow suppression occurred. Chimerism analysis (STR-PCR) from leukocytes on peripheral blood samples collected 24 hours after the NK-MSC infusion failed to detect donor-derived NK-MNC in the recipients (sensitivity: 3 %). Given these results, we conclude that long-distance transport of manipulated NK-MNC products between treatment and processing centers is feasible and reliable, clearly supporting the premise of PACT, and that CD3-depleted allogeneic NK-MNC from a MHC-mismatched relative can be safely administered to recipients of a recent autologous HSCT.This project has been funded in part by the National Heart, Lung, and Blood Institute, National Institutes of Health, under the University of Minnesota Contract [] N01-HB-37164 and HHSN268201000008C*PACT: Production Assistance for Cellular Therapies Disclosures:No relevant conflicts of interest to declare.

Jean‐Marc Reyrat (29/04/1967–28/10/2009)

Jean‐Marc Reyrat (29/04/1967–28/10/2009)

Discovery of the hepatitis C virus

After nearly 6 years of intensive investigations between 1982 and 1988 in my laboratory at Chiron corporation, in which numerous molecular biological methods were used to investigate the viral aetiology of parenterally transmitted non-A, non-B viral hepatitis (NANBH), a single cDNA clone (5-1-1) was isolated that was shown to be derived from a new flavi-like virus, termed the hepatitis C virus (HCV). After screening hundreds of millions of bacterial cDNA clones derived from different liver and plasma samples obtained from experimentally infected chimpanzees, a single HCV clone was eventually isolated using a novel, blind immunoscreening method in which antibodies derived from a clinically diagnosed NANBH patient were used to identify a cDNA clone encoding an immunodominant epitope within HCV nonstructural protein 4. Its viral origin was demonstrated by its specific hybridization to a large single-stranded RNA molecule of approximately 10,000 nucleotides found only in NANBH-infected samples that shared distant sequence identity with flaviviruses. Further, HCV clone 5-1-1 was shown to be extrachromosomal and to encode an antigen eliciting antibody seroconversion only in NANBH-infected chimpanzees and humans. Subsequent work demonstrated that HCV was the principal cause of parenterally transmitted NANBH around the world, with an estimated 170 million global carriers and that blood screening tests detecting circulating HCV antibodies and viral RNA could effectively eradicate the transmission of transfusion-associated NANBH. Key viral-encoded enzymes essential to its life cycle are now the targets of vigorous, ongoing drug development activities, and the feasibility of successful vaccination strategies has been demonstrated using the valuable chimpanzee model, without which any progress on HCV would not have been possible. My colleagues and coworkers who made essential contributions to the discovery of HCV were George Kuo, who had his own laboratory at Chiron and who provided intellectual and practical input, Dan Bradley of the Centers for Disease Control and Prevention, who provided a large supply of well-characterized chimpanzee samples and knowledge of the NANBH field, and Qui-Lim Choo, in my own laboratory, who provided many years of outstandingly dedicated and precise molecular biology expertise.

GM-CSF &amp; IL-2 ± autologous whole cell vaccine as adjuvant therapy in cutaneous melanoma: A phase II clinical trial

20027 Background: The results of various adjuvant trials in high-risk melanoma patients have demonstrated significant improvement in survival, but the results have been inconsistent. GM-CSF and IL-2 are 2 cytokines with distinct mechanism of action that are synergistic, and seemed logical to investigate as adjuvant therapy. Methods: Forty-five patients enrolled in this single-arm, open- labeled, IRB approved, phase II trial, to evaluate safety, tolerability and treatment effectiveness. After potentially curative surgery, each patient received 2-years of adjuvant therapy. The first year GM-CSF was administered as 125μgm/m2/day, subcutaneous (SC) for 14 consecutive days, followed by IL-2: 9 million IU/m2/day, SC for 4 days, followed by 10 days of no treatment. In addition, patients with resected large metastases, that yielded ∼100 X106 tumor cells, also received autologous whole cell vaccine, starting at the second GM-CSF cycle. The second year of treatment, each patient received GM-CSF only, 250μgm, SC twice/week. In event of resected recurrence, the same adjuvant therapy was re-administered. Eleven patients donated blood for dendritic cell (DC) counts. Results: This program was safe, well tolerated and compliance was excellent. Toxicities were mild to moderate and no hospitalizations were required. There was no increase in the number of DC during or after GM-CSF administration. Duration of follow-up ranged from 1 - 50 months, median 15.9. Thirty-two patients are alive 9/13 stage IV, 16/25 stage III, and 7/7 stage II (3B/4C). The survival data were expressed by Kaplan-Meier, and showed disease-free survival of .60 [95% CI], and overall survival of .64 [95%CI] at 21 months. There was no statistical difference in the survival by Log Rank between those who received biotherapy only vs. those treated by biotherapy and vaccine (p=.8). Conclusions: This adjuvant program seemed to be safe and effective, and such an effect does not seem to be mediated by DC. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration Berlex Oncology Chiron Corporation

Post-Consolidation Immunotherapy with Histamine Dihydrochloride and Interleukin-2 in AML: Long Term Follow-Up of Leukemia-Free Survival and Overall Survival.

PURPOSE. To assess the long-term outcome of AML patients (pts) in complete remission (CR) after a phase III study which compared the effect of post-consolidation immunotherapy with interleukin-2 (IL-2) and histamine dihydrochloride (HDC) on leukemia-free survival (LFS) and overall survival (OS).PATIENTS & METHODS. A multi-national, randomized, open-label, phase III study which recruited 261 AML pts in first remission (CR1) and 59 in subsequent remission (CR>1) was conducted between June 1998 and October 2000. Pts were enrolled after after termination of consolidation therapy, stratified by country and CR status, and randomized to either treatment or no treatment (standard of care, control). Forty-one % of pts were >60 yrs (median 57), and 54% were males. Study arms were balanced for known prognostic factors. Two patient populations were studied, i.e. all pts randomized (ITT population; n=320) and pts in CR1. The treatment was self-administered at home and included ten 3-week courses of low-dose IL-2 (aldesleukin, Chiron Corp) 18 000 U/kg sc bid, plus HDC (EpiCept) 0.5 mg sc bid. Cycles 1–3 comprised 3 wks of treatment and 3 wks of rest, whereas in cycles 4–10 the rest periods were 6 wks. As reported previously (Blood 2006; 108:88), the trial met the primary endpoint of prolonged leukemia-free survival (LFS) for all pts randomized (p=0.008 in favor of the treatment arm) at a median of 46 months of follow-up. The present long-term assessment of efficacy was conducted in August 2006, 30 months after the lock of the original data base.RESULTS. Follow-up forms were retrieved from 86% (n=107; 59 HDC/IL-2 recipients, 48 no treatment,) of the 124 pts pts who were alive at original study cut-off. In the long-term analysis, median LFS in the ITT population was 11 and 8.8 months in HDC/IL-2 and no treatment groups, respectively. In the CR1 population, median LFS was 15 and 9.7 months in HDC/IL-2 and no treatment groups, respectively. The benefit of treatment both for the ITT population (p=0.017) and in CR1 pts (p=0.026) was demonstrated by log-rank testing. Kaplan-Meier (KM) estimates for LFS at 60 months revealed a trend in favor of the treatment group in the ITT population (29.6 vs 20.6%; p=0.065) and a significant difference in the CR1 population (34.4 vs 21.7%, p=0.024). The median OS of CR 1 pts were 44 months (HDC/IL-2) and 28 months (no treatment), but the difference did not attain statistical significance (log rank test: p=0.22; KM estimate at 60 months: p=0.07).CONCLUSION. Post-consolidation immunotherapy with HDC/IL-2 results in significant long-term improvement of LFS.

Impact on Quality of Life of Postconsolidation Immunotherapy with Histamine Dihydrochloride and Interleukin-2 in Acute Myelogenous Leukemia.

PURPOSE: To assess the impact on Quality of Life (QoL) of postconsolidation immunotherapy with histamine dihydrochloride (HDC) in combination with interleukin-2 (IL-2) in acute myelogenous leukemia (AML).PATIENTS & METHODS: A multi-national randomized open-label phase 3 trial recruited 320 AML patients between 1998 and 2000. Patients were enrolled after termination of consolidation therapy, stratified by country and remission status (first or subsequent remission), and randomized to HDC/IL-2 treatment or no treatment (standard-of-care, control). The treatment was self-administered at home and included ten 3-week courses of HDC (0.5 mg sc bid; EpiCept Corporation) plus low-dose recombinant IL-2 (aldesleukin, 18,000 U/kg sc bid; Chiron Corp). Cycles 1–3 comprised 3 wks of treatment followed by 3 wks of rest, whereas in cycles 4–10 the rest periods were 6 wks. As reported previously (Blood 2006; 108:88), the trial met the primary endpoint of prolonged leukemia-free survival (LFS) for all patients randomized (P=0.008). Assessment of QoL, a secondary trial endpoint, was performed using the validated EORTC-C30(v2) instrument at 8 pre-defined study visits: baseline; pre- and post-cycles 3, 5, and 8; and at 18 months. Data were analyzed as specified in the EORTC scoring manual for 9 scales (1 global, 5 functional, 3 symptom scales) and 6 single-symptom items. Comparisons were made between and within groups at1.baseline vs. each visit,2.pre- vs. post-treatment, and3.baseline vs. end of treatment (18 months).RESULTS: At least one questionnaire was completed by 285 patients (89%). Patients in both study arms increased or maintained QoL status from baseline to last evaluation with respect to global health status, physical, cognitive and role functioning, and financial issues, as well as symptoms of fatigue, nausea/vomiting, pain, diarrhea, and dyspnea. Comparisons of pre- vs. post-treatment QoL assessments showed transient increases in fatigue (P=0.008), nausea/vomiting (P=0.006), and appetite loss (P=0.005) in the treatment arm.CONCLUSION: Post-consolidation immunotherapy with HDC/IL-2 self-administered at home by AML patients was associated with transient gastrointestinal symptoms and fatigue, but did not prevent recovery from induction/consolidation chemotherapy.

Prevalence of Elevated Alanine Aminotransferase Among US Adolescents and Associated Factors: NHANES 1999–2004

Nonalcoholic fatty liver disease (NAFLD) is a common cause of liver disease in children and adolescents. The majority of studies of NAFLD in children have been in select populations of the clinically obese. Study aims were to estimate the prevalence of elevated alanine aminotransferase (ALT, as a marker of NAFLD) in a general contemporary adolescent population and to identify leading risk factors for ALT elevation (>30 U/L). We analyzed data of adolescent participants (aged 12-19 years; N = 5586) in the National Health and Nutrition Examination Survey 1999-2004, a representative sample of the civilian noninstitutionalized US population. The prevalence of elevated ALT levels (>30 U/L) was 7.4% among white adolescents, 11.5% among Mexican American adolescents, and 6.0% among black adolescents. Elevated ALT levels were prevalent in 12.4% of male subjects compared with 3.5% of female subjects. Multivariable associations with elevated ALT levels were found for sex (odds ratio [OR] male vs female, 7.7; 95% confidence interval [CI], 3.9-15.1), ethnicity (OR black vs white, 0.6; 95% CI, 0.3-1.3; OR Mexican American vs white, 1.6; 95% CI, 1.0-2.6), waist circumference (OR per 1 SD, 1.4; 95% CI, 1.0-2.0), and fasting insulin level (OR per 1 SD, 1.6; 95% CI, 1.2-2.1). Age, C-reactive protein levels, and triglyceride levels were also positively and socioeconomic position inversely associated with elevated ALT levels. The magnitude of associations were similar across ethnic groups. ALT level is associated with waist circumference and insulin resistance even in a young population. These characteristics could be utilized to identify adolescents who may benefit from screening for NAFLD, offering an opportunity to prevent disease progression at an early age.

HCV-Specific T-Cell Response in Relation to Viral Kinetics and Treatment Outcome (DITTO-HCV Project)

The second slope of viral decline induced by interferon treatment has been suggested to be influenced mainly by the hepatitis C virus (HCV)-specific T-cell response; however, this hypothesis needs to be validated by results derived from experimental studies. To address this issue, the HCV-specific T-cell response of 32 genotype-1-infected patients of the 270 patients enrolled in the dynamically individualized treatment of hepatitis C infection and correlates of viral/host dynamics phase III, open-label, randomized, multicenter trial was studied in relation to viral kinetics and treatment outcome. Greater proliferative responses by HCV-specific CD8 cells were found before treatment in patients with a fast viral decline and with a sustained viral response. However, no significant improvement of HCV-specific CD8 responses was observed in the first weeks of therapy in both rapid viral responder and non-rapid viral responder patients. A mild enhancement of proliferative T-cell responses and a partial restoration of the cytotoxic T-cell potential was expressed only late during treatment, likely favored by HCV clearance. Early restoration of an efficient T-cell response does not seem to be an essential requirement for a rapid viral decline in the first weeks of treatment. However, patients presenting a better HCV-specific CD8 cell proliferative potential at baseline are more likely to present a rapid and sustained viral response. Therefore, future treatment protocols should consider the development of strategies aimed at improving HCV-specific T-cell responses.

Vaccine trials guide pandemic plans

Federal officials have stockpiled the equivalent of about 7.5 million doses of vaccine against an early isolate of influenza H5N1 and hope to use the results of scientific studies to parlay the bulk material into as many as 40 million doses. “This is a dynamic program,” Robin Robinson of the Department of Health and Human Services (HHS) Office of Public Health Emergency Preparedness said June 28 at the Conference on Antimicrobial Resistance in Bethesda, Maryland, an annual event sponsored by the National Foundation for Infectious Diseases. “Clinical results have really shaped our policy,” he said. Most pandemic-related research revolves around influenza H5N1, a strain that was first known to infect humans in 1997, went underground, then re-emerged in 2003. From then through late July the virus has been confirmed in 232 people worldwide and caused 134 deaths, most of them in Vietnam and Indonesia. Production of H5N1 vaccines for humans began in 2004, after HHS awarded contracts to Sanofi Pasteur and Chiron Corporation to produce what Robinson called a “prepandemic vaccine.” Although this formulation is not expected to be a good match for the precise influenza strain that eventually sparks a long-predicted pandemic, HHS hopes the vaccine, which is based on a human H5N1 isolate obtained in 2004 in Vietnam, will provide some protection.

Natural history of metastatic melanoma patients with CNS metastases

8054 Background: Melanoma has a high rate of CNS metastasis (mets). The objective of this study was to evaluate the characteristics and outcomes of melanoma patients (pts) who develop CNS mets. Methods: 333 pts with a diagnosis of CNS mets were identified from databases of 743 chemotherapy naïve metastatic melanoma (MM) pts enrolled on clinical trials between 1986 and 2004. Their clinical and pathological characteristics were reviewed. Results: The site of primary melanoma was head or neck (60/333; 18%), trunk/abdomen (144/333; 43%), limbs (66/333; 20%), ocular (2/333; 1%), or unknown (61/333; 18%). Median Breslow thickness (BT) = 2.2 mm; BT &lt; 1 mm = 39/217 (18%); and BT &lt; 2 mm = 95/217 (44%). Median Clark level (CL)= IV; CL I = 0/180 (0%); CL II = 14/180 (8%); CL III = 70/180 (39%); and CL IV = 96/180 (53%) . The median interval from diagnosis of primary melanoma to CNS mets = 29.6 (range 0.3–393) months (mos). Median survival (MS) from CNS diagnosis = 4.6 (range 0–120) mos. MS was highest for pts with brain mets (n=307; 4.8 mos) compared to pts with brain mets plus leptomeningeal disease (LMD) (14; 2.0 mos) or pts with LMD alone (11; 1.2 mos) (p=.0048 for pts with LMD vs. without). MS varied for pts with 1 (6.6 mos), 2 (4.2 mos), 3 (5.9 mos) or &gt;3 (3.5 mos) brain lesions at diagnosis of CNS mets. Among pts diagnosed with CNS mets at or prior to systemic therapy, MS was longer for pts with CNS mets only (n=20; 14.3 mos) compared to pts with CNS mets concurrent with extracranial mets (63; 7 mos) (p=.003). Patients who developed CNS mets after starting chemotherapy for extracranial mets (n=250; 3.7 mos) had a shorter MS than those diagnosed at or before systemic therapy (83; 7.9 mos, p&lt;.001). Among pts diagnosed after starting systemic therapy, CNS mets were detected ≤ 12 months from the start of chemotherapy in 30% of pts (MS = 3 mos), 12–24 mos in 37% of pts (MS = 4.6 mos), and &gt; 36 mos in 32% of pts (MS = 11.1 mos, p=.044 vs. other groups). Conclusions: This study represents one of the largest cohorts of pts with melanoma CNS mets. The presence of LMD, or development of CNS mets after starting systemic therapy, is associated with a worse prognosis. Among pts diagnosed with CNS mets at or before starting systemic therapy, the presence of concurrent non-CNS mets also portends for a worse outcome. Supported in part by Carol Courtney Memorial fund and Chiron Corporation. No significant financial relationships to disclose.

Identification of prognostic factors predictive of CNS metastasis in patients (pts) with advanced unresectable metastatic melanoma (MM)

8060 Background: Diagnosis (Dx) ofCNS metastasis (met) in MM is associated with poor prognosis. The objective of this study was to find prognostic factors that may predict CNS met. Methods: 743 chemotherapy (CRx) naïve MM pts were treated on 12 clinical trials during the past 15 years at MDACC. Demographics, tumor stage, and Rx outcomes were collected. Seven prognostic factors likely to predispose pts to CNS met were assessed: presence of liver met, elevated serum LDH, low serum albumin, site of primary, Clark level ≥ IV / Breslow thickness (BT) &gt; 3.5 mm, M1 stage, and number of visceral met sites. Time to event analyses considered the time from unresectable stage III/ IV disease to Dx of CNS met, or last follow-up if noCNS met. Prognostic factors were examined separately using a Cox proportional hazards model, with time from Dx of primary melanoma to Dx of unresectable stage III/ IV disease also included as a covariate. Estimates of CNS met-free curves were calculated using the Kaplan-Meier product-limit method. The incidence of CNS met between CRx-sensitive and CRx-resistant groups was assessed by Chi-square test. Results: CNS met (n=333) and CNS met-free (n=410) pt groups were similar in median age, gender distribution and primary M sites. Compared to CNS met-free pts, CNS met pts were more likely to have M1b + M1c advanced disease stage (M1b+c 291/333 (87%) vs. 286/410 (70%); p&lt;0.0001) and ≥ 2 visceral met sites (138/333 (41%) vs. 126/410 (30%), p=0.003), as well as greater BT (median 2.3 vs. 1.5 mm). Pts with elevated serum LDH had significantly shorter CNS met-free interval those with normal serum LDH (p=0.0009). CNS met-free interval was also significantly shorter for M1b (p=0.028) and M1c pts (p=0.05), but not M1a (p=0.8) pts, compared to unresectable stage III pts. Compared to pts with soft tissue/l. node disease, CNS met-free interval was significantly shorter for pts with 1–3 met sites (p&lt;0.0001), but not for pts with ≥ 4 sites (p=0.03). The incidence of CNS met was not significantly different between CRx-sensitive and CRx-resistant pts (p=0.2821). Conclusion: Prognostic factors predictive of high risk CNS met were identified that could be used to individualize Rx for MM pts. Supported in part by Chiron Corporation and Carol Courtney Memorial Fund. No significant financial relationships to disclose.

Meeting the need for treatment: the initiatives

Many diseases prevail because are old and ineffective, or simply do not exist. Tuberculosis (TB) is a major killer in poor countries, but no new anti-TB drug has been developed since the 1960s. There are rare, often life-threatening diseases, defined in Europe as affecting fewer than one in 2000 people, for which more orphan drugs need to be developed. There are diseases that are highly prevalent, such as HIV/AIDS in southern Africa, but while new, effective medicines exist, millions of people and their governments cannot afford them or they are not available at all. Neglected tropical diseases, such as malaria, affect millions of people, but most are too poor to constitute a market that is lucrative enough to justify drug research and development (RD 46 such projects were in the pipeline in 2005, according to a Wellcome Trust report. The generic industry provides cheap copies of brand-name medicines, has pushed down drug prices overall through competition. Access-to-medicines campaigners and generic companies argue that patents are a major barrier to meet the huge demand for affordable drugs, vaccines and diagnostics in developing countries. In contrast, the R&D-based pharmaceuticals industry, which comes up with innovations and patents them to recoup the R&D costs, argues that the bottleneck is not the fault of patent restrictions, but the lack of funds to buy these products and inadequate infrastructure to deliver them. Generic industry The global generic pharmaceuticals industry produces cheap copies of patented medicines for diseases such as HIV/AIDS, often pushing down the prices of the original patented medicines in the process. India is a major exporter of generic medicines to other developing countries. Of the more than 60 000 HIV/AIDS patients in nearly 30 countries in Medecins Sans Frontieres' projects, 84% receive generic medicines made in India. The generic industry helped establish the US Orphan Drug Act of 1983 to promote the development of medicines for small patient populations by providing incentives, such as tax benefits and exclusive marketing protection. Some say this model could be used to promote the R&D for for neglected diseases. The generic industry sees several threats to its business model, such as evergreening by pharmaceutical companies to extend their patent rights and data exclusivity, i.e. denying the release of information required to advance science. Public-private partnerships Public-private partnerships show that costly drug development can be combined with social responsibility. Under a 2002 agreement with US pharmaceuticals company Chiron Corporation, the Global Alliance for TB Drug Development obtained exclusive world rights to the compound, PA-824, and its derivatives, which may be developed into a new TB medicine. Chiron agreed to make the compounds royalty free in endemic countries, while retaining the right to develop and commercialize them for non-TB indications. Another example is a 2004 agreement between the International Partnership for Microbicides and Tibotec Pharmaceuticals, a Belgian subsidiary of pharmaceutical company Johnson & Johnson, to develop a microbicide to protect women from infection with HIV. …