The biotechnology industry has suffered serious setbacks in 1993. A drop in stock prices for the biotechnology industry has resulted from several blows to the industry: test results and the political climate. A successful biotechnology industry is necessary, however, for the treatment of human disease, competitive agriculture, and efficient management of the environment. The biotechnology industry has developed with controversy since it's inception in the 1970s. Recombinant DNA technology, patenting life, and patenting DNA sequences have all been monumental decisions for society and its laws which were induced by the emergence of the biotechnology industry. The recent setbacks in the industry provokes a reassessment of the industry in the United States of America in order to maintain a fertile biotechnology industry and the potential for profitability of investment in biotechnology.
GOALS OF MODERN BIOTECHNOLOGY
Manipulation of biology is desired to produce new drugs for the treatment of human and animal disease, better agricultural products, and a better living environment. Examples of new desired products include: new drugs to treat and prevent infectious diseases, new drugs to treat cancer and aging, new and improved agricultural products, new pesticides, and new products to control environmental insults.
STRUCTURE OF THE BIOTECHNOLOGY INDUSTRY
Genentech, the founder company of modern biotechnology, has grown to one of the largest of these firms with the development of recombinant human insulin. Robert Swanson and Herbert Boyer began Genentech in the early 1970s and it now has the "largest center devoted to biotechnology", an $85 million facility." Genentech was begun on the premise that "there was money to be made in recombinant DNA". Genentech invests 50% of its revenues on research and development compared to the average for the pharmaceutical industry of 16.7%. The price of Genentech stock has fared well during the recent slump in the prices of biotechnology stocks, actually increasing. It has developed: recombinant human growth hormone (BioTropin), human recombinant insulin, transforming growth factor-▀-1 (TGF-▀1), Activase to prevent blood clotting which has decreased death rates by 25.9% in heart attack patients, monoclonal antibodies to the HER-2 oncogene of breast cancers, _-Interferon (Actimmune has FDA approval for the treatment of chronic granulomatous disease), Pulomzyme (DNAse for the treatment of cystic fibrosis), a version of soluble CD4 for acquired immunodeficiency syndrome (AIDS) treatment, insulin growth factor-1 (IGF-1), recombinant tumor necrosis factor and others. A significant new product that they have recently unveiled is a genetically modified human recombinant insulin that can be processed by any cell in the body. This will lead to novel gene therapy approaches to the treatment of diabetes and other disorders.
Several organizations protect the interests of the biotechnology industry. The Biotechnology Industry Organization (BIO) has recently been formed by the merger between the Industrial Biotechnology Association (IBA) and the Association of Biotechnology Companies (ABC). BIO has taken the 500 employees from the Industrial Biotechnology Association and formed this organization of biotech companies, educational institutions, and state biotechnology centers. Dr. G.K. Raab, president and CEO of Genentech, is now the head of this organization as the elected chairman of the board. The president, Carl B. Feldbaum, will involved in battles with BIO against environment protection groups over issues of food labeling and the use of new plant varieties. This organization will serve to protect the interests of the biotechnology companies. These interests will include concerns about the patenting process and patent infringements. The ABC represented the interests of smaller biotechnology companies and these two organizations have differed on policy. They differed on legislation for orphan drugs, for example. The Pharmaceutical Manufacturers Association (PMA), which has some members in common with BIO, is another interest group of biotechnology.
The biotechnology industry has recently been given competing stock indices on the AMEX and the CBOE. On October 9, 1992 the Chicago Board Options Exchange (CBOE) and the American Stock Exchange (AMEX) simultaneously launched competing index options to track the volatile biotechnology industry. The Amex's (BTX, biotechnology stock index) is made up of 15 stocks and the CBOE's (BGX) is made up of 20 stocks. The overlapping stocks of these indices and the complete list of the AMEX's index are listed in appendix A. The AMEX's (BTK) is "equal dollar weighted" to avoid dominance of any one stock. At inception, it had a market capitalization of about 16.8 billion. The CBOE's is "price weighted", like the Dow, and at inception had a market value of about 19.2 billion.
Because of the discovery nature of the biotechnology industry, it has induced monumental changes in patent laws and the patent process. Since its inception in the 1970s, these changes have occurred in: the patentability of a discovery, the patenting process, the patentability of life, and orphan drug act legislation. The Intellectual Law Association and the organizations of the biotechnology industry (which includes BIO) protect the interests of the patents. The patentability of genetically engineered organisms was upheld in Diamond v. Chakrabarty in 1980 and transgenic plants and animals have since been deemed "patentable". Harvard University received the first patent for a genetically altered mouse, the "oncomouse", in 1988. The license was sold to Dupont. This has allowed the rapid development of recombinant DNA in biotechnology. The patent process is still in upheaval, however, several issues are still not settled, first to invent versus the "first to file" and the usefulness of an invention. In the United States of America, most university scientists publish first and file for a patent application later. This is not true in most countries. In Europe and most foreign systems, once a discovery is published, the inventor loses the right to a patent. An invention must be "useful, novel, and non- obvious" in order to be patentable. Novelty in United States patent law is: "An invention not known or used by others in USA or described in a printed publication...before the invention date and not described in a printed publication or in public use or on sale in the United States of America more than one year before the United States application date". In such a case, the patent obtains a 17 year monopoly for the invention. The United States of America has been negotiating a universal patent agreement with the with the World Intellectual Property Organization with a move toward the "first to file" rather than "first to invent" principle. In a recent case, however, the "first to invent" rule was upheld. On February 17, 1993, the U.S. Patent and Trademark Office determined that claims in a Regeneron patent application involving human ciliary neurotrophic factor (CNTF) protein and their pharmaceutical compositions containing it were allowable since they were the first to make the invention. This is despite the fact that Synergen was the first to receive a patent for CNTF in May 1991. CNTF is now in clinical trials for treatment of Lou Gehrig's disease. In the last couple of years, there has been a dispute about the patentability of DNA sequences. The National Institutes of Health (NIH) sought to patent DNA sequences that were found with expression tags. This would allow NIH to patent DNA sequences for proteins with unknown functions. If NIH had won this patent, it was argued that it would stifle future research on those genes. A similar case is under scrutiny yet. Researchers at Stanford want to patent sequences of the T cell receptor. This is because mutations in these genes may lead to autoimmunity, rheumatoid arthritis and multiple sclerosis. They seek to patent only specific uses, however, and the problem of ownership is therefore avoided. The Orphan Drug Act is a recent addition to laws brought about by the modern biotechnology industry. The United States Orphan Drug Act was established in 1983 to promote the development of drugs for rare diseases. Fifty drugs have since been approved for 59 conditions. There are still a number of scientific and legal aspects of this act which need to be refined. But this act will allow no other patents on the same structure for the same condition for seven years. If it is of a similar structure, it must be shown to be better in clinical trials.
The drug evaluation process is under currently under scrutiny in the United States of America as well. The method of drug evaluation depends upon the organism for which the drug is being developed. Drugs to be utilized for humans undergo pre-clinical trials and phase I, II, and III trials. Pre-clinical trials decide upon the efficacy and safety of the approach of the drug and are based upon animal and in vitro studies. Clinical trials occur sequentially with evaluation after each trial: Phase I trials establish that a drug is safe for human use on healthy volunteers, Phase II trials establish that the drug is effective and what dose is effective with a small number of patients, and phase III trials establish the dose and the effectiveness of the drug on a large number of patients. The data from the clinical trials are then submitted to the FDA for approval. A suggestion has been suggested to improve the phase II studies and discontinue or reduce the size of phase III clinical trials. This would decrease the time required for the commercial development of products for therapeutic use, which is now an average of eight years. Risk models for the evaluation of patients are important for the outcome of clinical trials of many of the newer biotechnology drugs. An example of the use new methods for the analysis of clinical data is a risk model based upon the APACHE III(TM) database. It is being used to monitor patients with systemic inflammatory response syndrome (SIRS). Cortech, Inc. is using this model in Bradycor clinical trials and describes the model as: "..the state of the art analytical tool for evaluating the benefits of sophisticated drugs in complex diseases". The APACHE III model, allows data on patients to be to analyzed according to their individual risk of mortality. It is accepted throughout the world for scoring the severity of illness in critically ill people. The APACHE method allows one to find patient subgroups according to their risk profiles. Statistical methods such as the APACHE III method increase the power of a test. This means that they are more likely to find a difference between the treatment and the control groups. Therefore, the subgroup which can benefit most from a drug can be determined. In other words, a drug which is effective in the treatment of diseases may be found ineffective by some statistical methods if the power of the test is poor. The Apache III (TM) risk model was derived from the analysis of over 100,000 patients within the APACHE III International Database.
CURRENT INTERESTS OF THE BIOTECHNOLOGY INDUSTRY
Modern biotechnology has made significant contributions to new methods of drug discovery, production, and delivery. New drugs include the new antibodies, soluble receptors, recombinant proteins, peptides, nucleic acids and derivatives of nucleic acids. These are used for treatment and for diagnosis of disease, the improvement of the environment, and better agriculture methods.
HEADINGS:
Drug discovery
Diagnosis
Antibodies
Immunology: vaccines,allergies, and transplantation,
Interleukins and other cytokines
Nervous system disorders
Cancer therapy
Antisense
Transcription
Splicing
Cell Cycle
Virology
Cardiovascular drugs
Wound Healing
Drug production
Drug delivery
Agriculture
Biological Control
Marine Biotechnology
Environment
Drug discovery...................................#Drug discovery
Biotechnology requires compound discovery. Old discovery methods relied on screening of a large number of compounds for a desired bioactivity. Compound libraries or organisms are screened for the production of molecules based upon pre-determined desired criterion. This has been used for discovering antibiotics in the past, and still is uses by some companies for drug discovery. Screening organisms for useful drugs is still used by a number of drug firms. There are basically two approaches to this: (1) screening libraries of compounds or products of organisms for a desired activity and (2) The screening
of organisms known to be resistant to certain infections or growth inhibit other organisms. One of the most recent compounds discovered by screening a large number of compounds for a desired activity is taxol. Taxol has been approved for treatment of ovarian cancer and is being produced by Hauser of Boulder, Colorado for ovarian cancer. Drug screening technology itself is a large part of modern biotechnology. Corporations, such as Pfizer, Inc., utilize high throughput screens to find novel compounds as well. A more modern variation to this approach is being used by some companies by developing large peptide libraries which are screened for binding to a structure or for bioactivity. Affymax, Selectide, and Enzon all have examples of this approach. Recombinant Fab combinatorial libraries have been developed for this purpose as well. Fab fragment libraries created from B cell RNA have been cloned into phagmid libraries to be screened for pre-determined binding specificities (Stratagene). Isolation of drugs based upon an organism's traits, such as resistance to infection is also an active area of biotechnology. For example, the biological activity of squalamine was recently found this way. The dogfish, which is resistant to in utero microorganism infections produces squalamine. This compound has been very active against Streptococcus sp., Staphylococcus sp., Escherchia coli, and yeast. Other companies are offering drug screening technologies for custom profiles of biomolecules after they are discovered to determine the spectrum of activities for it. This is offered by Oncogene Sciences as the GEMS(TM) discovery system.
Techniques for the discovery of drugs have recently been
under scrutiny for patentability. The patenting of the polymerase chain reaction (PCR) by Cetus has been one of the newest discovery methods with great impact to biotechnology. Cetus has since sold the patent to Hoffman-LaRoche. PCR, is a reaction where complementary DNA oligonucleotides are used to clone sequences from DNA or RNA. A heat stable polymerase, taq polymerase, is used to amplify the sequence in between the areas where the primers have annealed. In this way, if one knows flanking sequences of genes, genes can be cloned right out of an organisms DNA or RNA with very little analysis and time required. This all takes place in a computer controlled apparatus made by Perkin Elmer. Cetus has allowed the use of PCR in research, but the availability of other polymerases (cheaper), has instigated a crackdown on this patent by Hoffman-LaRoche. Hoffman-LaRoche, the Swiss based pharmaceutical company, has initiated a suit against Promega, Corp. of Madison, WI. Promega was licensed to make and sell taq polymerase for a limited number of applications which did not include PCR. They were undercutting Perkin-Elmer's prices by up to %60. Other techniques which are likely come under such scrutiny in the future include methods for sequencing the human genome.
Genetic techniques for disease diagnosis play an extensive role in modern biotechnology. New technologies include: FISHing (Fluorescent in Situ Hybridization), PCR, and enzyme labeled gene tag reagents. With the new technologies available, specific translocations of genes involved in malignant transformation can be found with greater precision. Furthermore, specific disease- causing genes can be searched for and found in individuals for genetic counseling purposes. PCR techniques can now be used to specifically determine the presence of viruses in tissues providing a direct link to viral infection, rather than just an association of the change in antibody titer of patients or victims. This has been used recently for the Four Corners Flu that has occurred in the northern New Mexico-Arizona area. The Hantavirus has been found in victims. This virus is transmitted to them by rodents that live in the area. Genetic data for the presence of the virus was found by PCR in tissues from the victims and deer mice. Biotechnology companies are also developing genetic techniques for the diagnosis of the involvement of specific oncogenes in cancer. For example, Oncogene Sciences has developed a hybridization probe kit for chronic myelogenous leukemia and there
are kits available for other translocations as well.
Antibodies are used for a number of purposes in biotechnology. Two basic types of antibody preparations are utilized, polyclonal and monoclonal. Polyclonal antibodies, heterologous antisera, immune globulin, or hyperimmune globulin are produced by whole living organisms. Monoclonal antibodies are produced by the fusion of a myeloma cell with immune lymphocytes. The product is a cell line that produces only one antibody that binds a desired determinant. Antibodies are useful for a number of things in biotechnology. First, they are very useful for prophylaxis against known exposure to pathogens or toxins. MedImmune is developing a number of antibodies for this purpose including polyclonal antibodies for prophylaxis against respiratory syncitia virus (RSV) and cytomegalovirus (Cytogam) and a monoclonal antibody for AIDS (Medi488). North American Biologicals, Inc. has been given rights to HIVIG by Abbott, a polyclonal antibody preparation from health HIV-infected individuals. This will be used to attempt to prevent the spread of HIV to the infants of pregnant women in a trial to begin soon. One in five women transmit the virus to their infant in Utero or at birth. Genzyme, Inc. has entered into an aggreement with Univax Biologicals, Inc. to produce drugs for the treatment of diseases associated with cystic fibrosis. Neozyme II will be given the license and development rights for these drugs from Genzyme, Inc.. HyperGam + CF will be used to give passive immunity to Pseudomonas infections. Pseudomonas spp. are responsible for life-threatening infections in cystic fibrosis patients. Second, they play a very significant role in diagnostic reagents. This includes bacterial and viral pathogens as well as cancers. For example, kits have been developed recently with monoclonal antibodies that can detect ras oncoproteins. These proteins are often involved in the development of aggressive cancers and may be involved in as many as one fifth of all cancers. Third, monoclonal antibodies have been developed for the treatment of a number of disease conditions and are in various stages of development. These include antibodies to tumor necrosis factor (a cytokine involved in septic shock) and antibodies to target tumors with cytotoxic drugs or radiation for cancer therapy. Seragen is developing fusion toxins which target activated T cells via the IL 2 receptor. So far, this treatment will be used for rhematoid arthritis, certain cancers (chemotherap resistant lymphomas) type I diabetes, and HIV infection. Furthermore, idiotype vaccines (killed MBP-reactive T cells are used to immunize against MS) have shown some promise and are in phase I trials Most antibodies in use are products of animals other than humans, therefore, because of species differences in their amino acid sequences, they can be recognized by the immune system of another animal. This is why humanized antibodies are being developed by a number of companies, to avoid rejection of the antibody in humans by the immune system. Humanized antibodies are the result of the genetic engineering of antibodies of other species with a desired reactivity for human treatment or prophylaxis. They have been modified to be as similar to natural human antibodies, while maintaining the desired reactivity. Companies in clinical trials with humanized antibodies include: Cytogen, Genentech, IDEC Pharmaceuticals, Protein Design Labs, and Scotgen Biopharmaceuticals. New methods of achieving humanized antibodies include the use of mice which are transgenic for a mini-human antibody locus. This is being are being tried by GenPharm of Mountain View, CA. In this way,
these mice can be injected with a desired immunogen, and the monoclonal antibodies produced can be screened for only the antibodies of human sequence. They will not have to be modified by further genetic engineering because they already contain only human sequences. Lastly, antibodies and derivatives of them are being used in medical imaging. For example, they will allow better localization of areas for surgical removal of cancer tissue.
Vaccines are a large part of the biotechnology industry and remain the only method of combating many diseases, particularly viruses. The most celebrated vaccines are now the vaccines against the human immunodeficiency virus (HIV) virus. Both, vaccines aimed at preventing infection with the HIV virus and AIDS and novel vaccines aimed at quelling the disease after infection are being developed. Immune Response Corporation released the results of its first clinical trial of a novel vaccine designed to quell AIDS disease on June 13, 1993. A whole virus preparation was used in which the virus has been chemically stripped of its envelope. A study of 103 patients suggested that those who received the experimental vaccine maintained slightly reduced levels of virus. Although the results are promising, it is less of a response than is desired for an AIDS treatment. Some virologists have said that the differences were so slight that the results were impossible to interpret. Several biotechnology companies have taken the same approach to stimulate the cellular immune response with particular proteins of HIV. Furthermore, this vaccine may be used for the prevention of infections. It has been shown that a small proportion of people who have been infected with the HIV virus do not develop AIDS and their antisera never tests positive for HIV. Cellular immune responses can be detected, however. Alpha-1 Biomedical of Washington D.C. and Cell-Sci of Alexandria, VA are developing these vaccines. They are utilizing HGP30, the P17 core protein of HIV, which is expressed very early in HIV infection. The vaccine is actually being produced by Viral Technologies, Inc.. So far, clinical trials have detected no side effects for this vaccine. The most popular approach, and probably the most likely to succeed for protecting individuals from infection, will probably involve the stimulation of neutralizing antibodies or a combination of stimulating both cellular and humoral immune responses. If the HGP30 vaccine approach is utilized it is hoped that a combined vaccine will be used which stimulates both the cellular immune response and antibodies. Neutralizing antibodies are directed against gp120 (or gp160, an unprocessed viral peptide containing gp120). This is the protein that binds the CD4 molecule on T cells which the HIV virus uses to enter the cell. A number of companies are developing these vaccines. Mammalian versions of the gp120 vaccine are being developed for the LAI (French isolate first discovered) and MN strains of HIV by Genentech. This will have the post-translational carbohydrate additions of mammalian cells. A mammalian version of the SF-2 strain is being developed by Biocine/Chiron. MicroGeneSys is developing the gp120 and gp160-protein-based vaccines from the LAI strain in insect cells with baculovirus vectors. These cells produce a protein with some of the carbohydrates attached that mammalian cells add. The gp160 molecule is being used in an approach similar the above cellular immunity vaccine, HGP30. It is being used on HIV-infected pregnant women to prevent transmittance to the fetus. It is hoped that it will stimulate both cellular and humoral immunity and prevent the transplacental transfer of HIV to the fetus. This vaccine (VaxSyn) has also been approved by federal government for HIV-infected armed service personnel. Immuno AG has the version of this vaccine which is expressed in mammalian cells. Vaccines which stimulate the antibody response to prevent HIV infection can utilize the gp120 molecule or synthetic peptides of the gp120 molecule. Peptides which correspond to the V3 region of the gp120, a region which mutates frequently, are being developed by Repligen. A cocktail of a number of peptides corresponding to the sequence of the V3 regions of a number of HIV isolates is being developed which is hoped to induce immunity to the large number of mutants of HIV. Since these peptides are often not very immunogenic, powerful adjuvants will need to be used. One such adjuvant, Stimulon with QS-21, has been developed by Cambridge Bioscience. Genentech has chosen this adjuvant for use with its gp120 vaccine as well. A number of other modern vaccines are also being developed by the biotechnology community. One such approach utilizes the B subunit of cholera toxin which is a mucosal adjuvant. Mucosal immunity has been achieved for SIV (simian immunodeficiency virus) in monkeys. More "high-tech" approaches to vaccines are also being developed against the AIDS virus and other organisms. They involve the use of live vectors for delivering the immunogen. Recombinant proteins, such as gp160 or gp120 are being placed in Vaccinia vectors (modified Small pox viruses) or BCG (Bacillus Calmette-Guerin). These vectors stimulate both cellular and humoral immunity. Bristol-Myers Squibb has a version of this vaccine. It is a gp160-vaccinia recombinant vaccine. Other modern vaccines are being developed to a number of pathogens. MedImmune itself is developing a number of modern vaccines against AIDS, parvovirus, and Lyme disease. Abt Associates of Cambridge, MA and Family Health International of Research Triangle Park have been awarded this a 6.5 million dollar contract to test the AIDS vaccines.
Treatment of allergic reactions to a number allergens has been undertaken by modern biotechnology companies. ImmunoLogic Pharmaceutical Corp. is developing Fel D I as CATVAX and AllerVAX for the treatment of two common allergies, allergies to cats and ragweed. CATVAX is now in phase II clinical trials.
Transplantation is an important area of research in biotechnology today. Two of the most popular drugs in transplantation immunology, used for inhibiting the rejection of transplants, are cyclosporin A and FK506. These drugs can be taken orally and act by inhibiting the activation of T cells. This is accomplished mainly by the inhibition of a T cell transcription factor, NF-AT. Serious side effects still occur with these drugs, which include renal toxicity and neurotoxicity. Adhesion molecules are now in vogue today and a large number of biotechnology companies are devoted to studying these molecules. Newly discovered adhesion molecules have been targeted to inhibit transplant rejection. Adhesion molecules can be classified by their necessity for signaling by T cells the immune system, these cells are responsible for graft rejection and the initiation of many immune responses. The LFA 1, CD5, and CD2 molecules are not involved in cell signaling, but increase the avidity of the interaction of the lymphocytes with antigen presenting cells or targets. CD4 or CD8 molecules are involved in signaling as are the antigen specific receptors of T cells. A recent addition to adhesion receptors of T cells involved in signal transduction is the CD28 molecule. This molecule binds the B7 ligand and this interaction is necessary for the generation of cytotoxic T cells. Cytotoxic T cells are involved in the removal of virus-infected cells as well as the rejection of tumors and grafts. Molecules similar to CD28 which block the interaction of CD28 with B7 have been used for blocking allograft rejection. CTLA4Ig is one such drug that has inhibited graft rejection.
The treatment of autoimmune diseases is being targeted by modern biotechnology as well. New MS treatments are described in the section on nervous system drugs. Similar treatments are being considered for other autoimmune diseases such as rheumatoid arthritis. Seragen is developing new treatments for autoimmune diseases and HIV by targeting activated T cells via their IL 2 receptor with fusion toxins.
Biomedicine:Interleukins and other cytokines.....#
Interleukins are cytokines which allow intracellular communication between cells of the immune system. There are now 14 interleukins numbered 1-14. Other cytokines of current interest in biotechnology include: TGF-_, EGF, FGF, TGF-▀, TNF-_,TNF-▀, CNTF, BDNF, neuroleukin, NGF, IGF, interferons, and Stem cell factor (SCF).
The Interleukin 1s: Interleukin 1s are a family of structurally related cytokines that all bind to two different receptors. This family of proteins is composed of IL 1_, IL 1▀, and IL 1ra. These molecules are involved the initiation of immune response (proinflammatory) and in the acute phase response of the liver. Furthermore, receptors can be found in the neuroendocrine and cardiovascular system. It is one of the cytokines responsible for septic shock and is involved in the stress response and the secretion of glucocorticoids.
Interleukin 2: Interleukin 2 is the first T cell growth factor which was characterized, isolated, and cloned by recombinant DNA methods. For this reason, this cytokine has been under investigation for the treatment of immune disorders and to generated immune cells capable of killing cancer cells. This has worked to some extent with cells called "lymphokine activated killer" or LAK cells. Immune cells are treated in the laboratory with IL 2 and then readministered to cancer patients. Systemic use of IL 2 has proven to have too many side effects. Another approach to kill tumor cells has involved transfecting tumor infiltrating lymphocytes (TILS) with the IL 2 gene in the hope of delivering the IL 2 to tumors locally. These studies are being performed by the National Institutes of Health.
Interleukin 3: Interleukin 3, also known as multi-CSF (colony stimulating factor) promotes the differentiation of early T cell precursors, supports the establishment of T cells lines and supports the proliferation of a broad distribution of hematopoietic stem cells. Furthermore, it synergizes with a number of cytokines in colony formation by primitive stem cells (CD34+ cells).
Interleukin 4: Interleukin 4 was first characterized as a B cell cytokine; however, it has turned out to be an important T cell growth factor as well. It is involved in the growth of cytotoxic T cells and T cells that collaborate with B cells and it is absolutely required for IgE production. Anti-IL 4 antibodies have proven to be useful in some types of vaccination, skewing the development of T cell subpopulations: favoring T inflammatory (IL 2 producing) cells in mice. Furthermore, renal cancer has been treated in mice by the transfection of the tumor cells with the a functional IL 4 gene. Genetic Therapy, Inc. has produced a retrovirus which produces IL 4 for the treatment of melanomas, kidney cancer, breast cancer, and colon cancer.
Interleukin 5: Interleukin 5 is involved in B cell proliferation and differentiation. Furthermore, it acts as an eosinophil growth factor and is thus involved in parasite and tumor immunity.
Interleukin 10: Inteleukin 10 is produced by TH2 cells, which are responsible for the production of IL 4. It will inhibit the production of TH1 cells (IL 2 producing).
Interleukin 12: Inteleukin 12 is produced by macrophages in response to stimulus, such as Listeria. This cytokine is responsible for the development of TH1 cells. The loss of TH1 cells is now believed to play a role in AIDS.
Interleukin 13: Interleukin 13 inhibits the secretion of inflammatory cytokines in response to sepsis (See Interleukin 1). Furthermore, it synergizes with Interleukin 2 in the induction of interferon-_ production.
TNF: Cachectin and lymphotoxin are the tumor necrosis factors, TNF-_ and TNF-▀. TNF-_ is involved in septic shock and antibodies to it are being considered for treatment of septic shock. Gene transfection experiments are also being performed with this cytokine which are analogous to the IL 2 transfections. TILs are being transfected in the laboratory and transferred back to the patient.
TGF-▀s: TGF-▀s have pleotropic effects on many cell types including immune cells and fibroblasts. They are involved in immune memory, immune suppression and wound healing.
Transforming growth factor-▀s (TGF-▀s) (Genentech:TGF-▀1, Celtrix: TGF-▀2, and Oncogene Sciences:TGF-▀3), are members of a family of over 18 proteins. These cytokines are being developed to treat chemotherapy side effects, ischemic heart injury, and wound healing disorders.
Cytokine combinations: The use of combinations of cytokines for the purpose of treatment of disease or the growth of desired cells is being performed by a number of companies. Cell-Sci is one pioneering company which is doing this with "Multikine". Amgen has also been involved in this research. One cytokine combination which has proven useful for the purpose of growing stem cells (CD34+, see stem cells) is the combination of Stem cell factor (SCF) with other synergizing cytokines such as G-CSF, GM-CSF, Erthyropoeitin (EPO) and IL 3.
New drugs have been found for a number of nervous system disorders including, brain tumors, amyotrophic lateral sclerosis (Lou Gehrig's disease), and multiple sclerosis (MS). Betaseron, a form of ▀-interferon (IFN-▀1b), has recently been approved for use by Chiron Corp. as the first licensed treatment for MS. It is the only drug "specifically approved" for the treatment of MS. The drug is being produced by Berlex, a unit of Schering AG (SCHG.F). MS affects 250,000 to 300,000 Americans. MS is mediated by T cells which react with the myelin basic protein (MBP) of the central nervous system. ▀-interferon is produced by suppressor T cells and it is therefore hoped that the administration of Betaseron to MS patients will suppress the MBP-reactive T cells. This drug is not a cure and other more effective drugs are needed. However, it has been claimed that this drug is as effective as other drugs at controlling the disease. The administration of 8 million units of Betaseron every other day decreased the relapses and the number of patients with detectable brain lesions compared to placebo controlled "relapsing-remitting MS". In the one study performed so far, twenty nine percent of the placebo patients had brain lesion detectable by Magnetic resonance imaging (MRI) while only 6% of the Betaseron treated had the lesions. Furthermore, the number of attacks was decreased, 0.9 vs 1.31 per year. Other drugs, specifically targeting the MBP-reactive T cells are being developed and will probably be more effective. Drugs such as toxin-conjugated antibodies which target the activated T cells and the T cells with the receptors specific for MBP are being developed by a number of companies. The induction of oral tolerance to MBP by feeding patients the whole protein has also shown great promise (Selected reading, Weiner et al.). Similarly this is being tried for rheumantoid arthritis by feeding individuals collagen.
One obstacle to treatment of neurological disorders has already been overcome. Getting drugs across the blood brain barrier, BBB. The BBB does not allow large molecule to pass across it. T cells get there only because they are activated and are able to induce their transport across the barrier, transiently. Several companies, Alkermes and Cambridge Neuroscience, have been able to target receptors at the BBB with antibodies and have gotten drugs to pass into the brain. Therefore, drugs such as antibodies, cytokines or peptides may be targeted across the BBB where they can be effective. One cytokine which may be used for the treatment of neurological defects or for the prevention of neurological defects due to ischemic injury is nerve growth factor, NGF. NGF has been shown to protect neurons from apoptosis (a form of programmed cell death). Nerve growth factor is also being explored for its ability to inhibit neuron death in the case of brain trauma. Cephalon produces NGF as a nerve protector and has been targeting the BBB as well.
Molecular surgery for the treatment of brain tumors may be possible soon as well. Retroviruses with the Herpes Simplex Virus thymidine kinase gene (HSV-tk) were introduced into rat brain tumors by HSV-tk-transfected fibroblasts. The rats were then treated with a drug that will kill a cell containing the HSV-tk gene, gancyclovir. Complete tumor regression was observed.
New drugs are being produced by biotechnology for the treatment of peripheral nerve abnormalities as well. Ciliary neurotrophic factor (CNTF) is now in clinical trials for amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease). Regeneron is now finishing phase II clinical trials. This protein drug has been shown to inhibit apoptosis (programmed cell death) in neurons. Interestingly, the gene responsible for familial ALS has been shown to be the Cu/Zn superoxide dismutase gene. The product of this gene protects cells from radicals. Free radicals are chemical agents that induce damage in the cells molecules, such as their DNA. Furthermore, a protein which is believed to monitor DNA integrity, p53, is involved in the induction of apoptosis. It may be that CNTF will inhibit the apoptosis induced by this gene by this mechanism. Phase II clinical trials for CNTF have been disappointing, in most cases showing no differences between treatment and control groups. Although it has been shown to be safe, antibodies to CNTF developed in all patients. Furthermore, some patients developed elevated temperature, diarrhea, cold sores, and anorexia. One group of patients patients in the Regeneron study may have shown a response to CNTF. Those given high doses (12 patients) demonstrated less deterioration of muscle strength (72%) and lung function (44-69%) than the group that received the placebo (14 patients). It should be pointed out that the original study was composed of 240 patients: one third was to receive 30╡g/kg of CNTF, one third was to receive 15╡g/kg, and one third was to recive a placebo. The phase III clinical trials are designed for a 95% probability of determinng if CNTF is effective. Synergen an Syntex are planning phase II/III studies of CNTF. So far, the side effects are the same as those observerd in the Regeneron study. BDNF (brain derived neurotrophic factor) is also currently being considered by Amgen and Regeneron for treating ALS. It has been shown to protect neurons as well. Other drugs which will decrease radicals in cells are also being explored. Vitamins C and E will do this as well.
Many new approaches to the treatment of cancer are under investigation by modern biotechnology. Drugs which kill dividing cells are the most frequently used today. These non-specific drugs have the problem of killing many normal cells as well as tumor cells. Most notably, they kill lymphocytes. Furthermore, tumor cells often become resistant to these drugs by the up- regulation of a multiple drug resistance gene. This leads to the overproduction of P-glycoprotein and the resistance to a large number of chemotherapeutic agents. Therefore, new approaches should be more specific for the cell type, will hopefully have less toxic side effects, and be less prone to multiple drug resistance. The goal is to be more specific and more efficient at killing tumor cells. This is being approached at several levels: cell surface growth factor receptors, intracellular second messengers for growth factor receptors, and the transcription of nuclear oncogenes, the introduction of normal tumor suppressor genes and new methods of immunotherapy. Drugs which are targeting the activation domains of tyrosine kinases are being developed by a number of the above mentioned methods. It is hoped that drugs which are very specific for the tyrosine kinases responsible for altered signaling in cancer cells will be found. For example, drugs which will target SH2 and SH3 domains of a cell-type specific tyrosine kinase would be more useful than drugs which inhibit any dividing cell. Drugs which inhibit Ras oncogenes are being studied by a number of companies. Most recently, Merck Research Laboratories in West Point, PA and Genentech, Inc. of S. San Francisco, CA, have been looking for drugs that inhibit the post-translational modification of the ras oncogenes. In order to function efficiently, the ras oncogenes require membrane association via a farnesyl group. Several custom drugs have been found by these groups which will act in low concentrations (micromolar). Furthermore, no toxicity has been found yet in vitro. Ligand Pharmaceuticals of San Diego, CA is another company developing small molecules with propriety technology which target intracellular receptors (IR) for hormone receptors. Targeting the ligands for growth factor receptors is being preformed by a number of biotechnology companies as well. Antagonists, soluble receptors, or neutralizing antibodies to the ligands which cause the aberrant growth of cells are being sought and developed. The aberrant production or control of transcription factors has been shown to cause the transformation of many cell types. Therefore, an approach with great promise is to find a method of inhibition of these aberrantly controlled transcription factors. Oncogene Sciences has made a commitment to this and have developed diagnostic reagents for translocations involved in leukemia and is focusing a large part of their company's effort on transcription factors. Furthermore, the have developed diagnostic reagents for mutations in the ras oncogene. This oncogene is involved in a number of aggressive malignancies.
Tumor suppressor genes are now a very active area of cancer research. These are genes which normally inhibit the growth of cells; the loss of their function causes transformation. The retinoblastoma protein (RBP), p105 and p53 examples of these genes. Transfection of transformed cells with a wild type p53 has been shown to cause the reversion of these cells to a normal phenotype. Onyx Pharmaceuticals is committed to the production of "small molecule therapeutics for cell growth and differentiation". This California company was formed in 1992 from the molecular oncology group of Cetus, Corp. and it is financed by three venture capital firms and Chiron, Corp. The will be specifically targeting the ras gene superfamily and tumor suppressor genes. Genetic therapy for lung cancer is being considered as well. A retrovirus containing the antisense k-ras gene or wild type p53 will be used to attempt the transfer of these genes to the tumor cells to regulate their growth.
New cancer treatments based upon immunotherapy have shown alot of promise recently. As mentioned above, some T cell adhesion receptors are involved in signal transduction. This is the CD28 molecule and the B7 ligand. This interaction is necessary for the efficient generation of cytotoxic T cells. Cytotoxic T cells are involved in the removal of virus-infected cells as well as the rejection of tumors and grafts. This molecule or modifications of similar B7 binding molecules (CTLA4Ig) have been used to prevent graft rejection and to generate cytotoxic T cells which cause rejection of tumors in mice.
Transfection of effector T cells with genes that encode the T cell receptors (TCR) which recognize tumor-specific antigens have demonstrated success in a limited number of cases. Specifically, a lymphoma caused by the Epstein-Barr virus (EBV). T cells from a sibling were transfected with the TCR genes and then administered to the patient. Other treatments are being developed for other tumors with the same approach: the targeting of viral tumor antigens. Targeting tumor specific antigens is also being considered for the treatment tumors of non-viral origin. Furthermore, processed peptides of oncogenes are being considered. The mutation of proto-oncogenes causes the formation of an oncogene and often a cancerous cell. This can be a "single hit" of a "multiple hit", i.e the result of one oncogene or the result of several oncogenes. Some oncogenes may be processed by the cell and present the "oncogene-specific" peptide to autologous T cells. For example, ras mutants have been considered. Vaccines are being attempted which will generate these T cells.
Stem cells are progenitors of mature cells in the organism. There are many different types of stem cells. These cells will be useful for replacement cell therapy and gene therapy. Stem cells of the immune system are the most noteworthy at this time. These cells are capable of generating many mature cell types of the immune system. Cellpro, Inc. of Bothwell, WA has been able to isolate hematopoietic stem cells. Using a monoclonal antibody that defines a determinant CD34 (cluster of differentiation number 34) they have isolated hematopoietc stem cells by positive selection with their patented CEPRATE system. The patients bone marrow cells are labeled with the anti-CD34 monoclonal antibody and are then passed over a column that will bind that antibody. The cells are then positively selected by washing them off of the column with an acid wash. This removes the cells from the column and the antibody from the cells. Applied Immune Sciences has a similar cell separation system, the AIS MicroCELLector. These stem cells have also been grown with the use of stem cell factor (SCF) in vitro. It has been found that the administration of these stem cells to cancer patients after treatment with drugs such as Cytoxan and Thiotepa with G-CSF (granulocyte-colony stimulating factor, Amgen's Neupogen) has significantly decreased their chemotherapy side effects. GM-CSF (granulocyte macrophage colony stimulating factor, Immunex's Sargramostim) is also under investigation for these treatments. A study is presently being performed with this protocol followed by taxol treatment for ovarian cancer. Similar treatments are in phase I and phase II clinical trials for non-Hodgkin's lymphoma and breast cancer. CD34+ cells have also been used for allogeneic bone marrow transplantation with good results. Cellpro, Inc. is expected to get half of this market. Future prospects for CD34+ cells include genetic engineering applications. If these cells are modified with a gene to deliver a drug, they may be present in the body for a longer period of time than more differentiated cells. TGF- ▀3 (see Cardiovascular drugs) is also being developed for cancer therapy. It will be used to decrease side effects caused of chemotherapeutic drugs, such as oral mucositis. Oral mucositis is a painful wound healing disorder.
Antisense drugs are some of the most promising drugs which are on the verge of an explosion in use. Prototypes of these drugs have been used to: inhibit gene function for the treatment atherosclerosis in animals, inhibit tumor causing retroviruses in mice, inhibit oncogene transcription, inhibit of virus production, inhibit pigment production in flowers, and inhibit the breakdown of tomatoes. Antisense compounds aimed at papilloma virus infections, cytomegalovirus (CMV), and herpes viruses are in several stages of development. Enzo biochem has an exclusive antisense patent with the State University of New York, but a number of companies are developing antisense drugs. Isis is now in phase I clinical trials for human papilloma virus infection. A disease thought to infect over six million women in the U.S.A.. Antisense compounds have been devised which inhibit gene transcription by triplex formation, mRNA translation, and cause the breakdown of specific mRNAs. Isis has also developed antisense compounds with their new patented "second generation" chemistry that inhibit the ras oncogenes, specifically c-Ha-ras. These antisense compounds were shown to be effective against cancer cells by researchers at Isis. Similar oligonucleotides are being developed by Microprobe of Bothwell WA. They are developing nucleotide and protein drugs for the detection and treatment of conditions such as vaginitis. Antisense compounds which have shown great promise recently have been developed by Gilead Sciences. C-5 propyne pyrimidine containing antisense oligonucleotides are ten times more potent than other antisense compounds. These drugs make up a significant proportion of the "designer" drugs of modern biotechnology.
The ability to specifically regulate genes will aid conditions such as: cancer, genetic disorders, and aging. Transcription inhibitors are being developed and are in the very early stages of development. Areas to be targeted for these compounds will include oncogene specific transcription factors of viral enhancer/promoters. Oncogene Sciences has made a commitment to study transcription inhibitors for anticancer therapeutics. A company that was formed recently, Tularik, Inc., will look for transcription inhibitors as drugs. Steven McKnight has formed Tularik, Inc. with Robert Tjian of Univ. of Calif., Berkeley and David Goeddel of Genentech. Tularik, Inc. will focus on inhibitors of transcription for these drugs. The first targets for drugs will be viruses such as herpes viruses. Drugs which enhance transcription of specific genes are also being developed. Vertex, Inc. has licenesed the use of small organic molecules from Children's Hospital Oakland which enhance the transcription of fetal hemoglobin. Fetal hemoglobin is not normally utilized by adult cells. It has a slightly higher affinity for oxygen than adult hemoglobin which aids the transplacental transfer of oxygen. The gene is not actively transcribed in the adult. These organic molecules, one administered orally and the other intravenously, potentiate the transcription of this silent gene in adults. These molecule will be used to treat the hemoglobin disorders, ▀-thalassemia and sickle cell anemia. Fetal hemoglobin has been shown to benefit these patients. Vertex, Inc. is committed to rational-drug design.
Splicing.........................................#Splicing z
Splicing is the process used by Eukaryotic cells to combine gene segments into a functional mRNA for translation. Eukaryotic genes are often separated on the DNA chromosome by "untranslated regions" called Introns. To join the translated regions (the Exons), the large RNA transcript is "spliced" to produce a functional mRNA for translation into protein. This process is being targeted by a number of companies to produce a number of products including better agricultural products and AIDS therapeutics. For example, the alteration of the normal splicing process could inhibit gene translation. Furthermore, the production of novel "spliceosomes" could destroy viral RNA, such as HIV RNA. Ribozyme Pharmaceuticals of Denver, CO, a biotech startup company of United States Biochemical Corp., is one company involved in this research. They currently have are working with DowElanCo to produce better agricultural products, such as a disease-resistant corn and a corn which produces more oil. Innovir Laboratories of New York, NY is cleaving target RNA with the enzyme RNAase-P attached to specific sequences. They are currently interested in blocking viruses and disease causing proteins.
The process of cell division, or proliferation takes discrete steps. Through an ordered progression of stages a cell divides. In these steps, the cell synthesizes new proteins, DNA, and other macromolecules. The cell cycle is most simply divided into the stages: G0 (G stands for gap), G1A, G1B, S (Synthesis, DNA), G2, and M (mitosis). Most cells of an organism are in the the G0 stage. Mitogens or growth factors move the cell out of this stage and into the G1A stage. It is at this step that many of the drugs aimed at transcription factors and growth factor- associated protein kinases are founded. See the section on cancer therapy. TGF-▀s inhibit cell division in late G1 stage. The retinoblastoma gene product and p53 are examples of tumor suppressor genes that act at the G1/S boundry. Mutation of these genes is associated with several malignancies. Gene therapies based upon the introduction of a non-mutated or wild type RB or p53 are being tried as a cancer treatment. See cancer therapy above. Other portions of the cell cycle are being targeted by a number of companies. See transcription above. One company specifically devoted to cell cycle-based therapeutics is Mitotix, Inc..
Antiviral compounds will be highly profitable for modern biotechnology and are under intense investigation. As already described, viruses are obligate intracellular organisms, they must replicate in living cell. This is because they require energy production from the cell as well as its macromolecule synthetic machinery. Since these functions are required by the cell as well, these functions are difficult to target for antiviral compounds because normal cell function is disrupted. An antiviral compound must be created which interrupts virus specific functions. This will depend on the type of virus and often on the specific virus. This is in contrast to anti-bacterial agents such as penicillin or streptomycin. These agents act on components specific for bacteria and will act on many different bacteria. The life cycle for a virus is: adsorbance, penetration, uncoating, replication and/or latency, packaging, and maturation and release. These are the stages that are targeted for antiviral compounds. Adsorbance is the step most often targeted. This is accomplished with the induction of antibodies by vaccines, as mentioned above. Other methods for blocking virus adsorbance include the recent use of soluble CD4 (sCD4) to block the adsorbance of HIV to T cells as an AIDS treatment. Replication is also targeted for antiviral compounds. This approach is dependent upon the type of virus. Retroviruses, RNA viruses which must integrate in to DNA to replicate, utilize a virus-specific enzyme for replication. This enzyme, reverse transcriptase, transcribes DNA from the virus's RNA. This is the target of AZT, ddI, and ddC. Although AZT has been shown to prolong life for AIDS patients, it has been disappointing that there is no difference in the patients who have been treated before ARC (AIDS- related complex). AZT apparently has a "limited benefit for a limited period of time". Furthermore, a recent panel of independent experts does not recommend AZT therapy to people with extremely low CD4 counts. Healthy individuals have CD4 counts ranging from 800-1200 and people infected with the HIV virus have CD4 counts in the 200-500 range. A new formulation of drugs which also targets reverse transcriptase has been devised recently by scientists at Massachusetts General Hospital that incorporates 3 reverse transcriptase drugs, U-90,152. It contains all three of the above mentioned drugs, AZT (zidovudine), ddI (dianosine) and ddC (zalcitabine). This is very potent in the laboratory. There was data to suggest that it may be difficult for the virus to mutate and become resistant to this combination. However, recent reports have claimed that HIV is able to mutate after 20-30 generations in the laboratory. This type of convergence therapy is also being developed by Merck for HIV. They are conducting clinical trials with a two-drug combination which utilizes the same principles. Antisense compounds are also being used to target viruses. These drugs can be specific for mRNAs of viruses and block the production of virus specific proteins or be targeted toward DNA of the virus and inhibit the transcription of specific mRNAS. Gene therapy approaches may prove to be fruitful here as well, and some adeno-associated virus vectors with antisense HIV genes have been tried and have been shown to work in vitro. Genta has an antisense drug for the treatment of acute leukemia caused by the HTLV I virus. This has been shown to work in mice. Isis has an antisense drug for human papilloma viruses and others are being developed for HIV, CMV, and Herpes viruses. The antisense drug for human papilloma virus is in pivotal phase II clinical trials. A patent has also been issued to them to them for Epstein-Barr virus infections. A herpes virus that infects over 80% of the worlds population. This antisense drug is directed at EBNA-1, a gene that is expressd in some lymphomas, leukoplakias, and carcimonas. Gilead Sciences, Inc. has a new powerful antisense drug which is ten times more effective than those previously used. Abbott Laboratories of Illinois have developed a drug for treatment of HIV infection which targets the maturation step of the virus. HIV requires a dimer of a protease molecule for maturation. Abbott Laboratories has designed a symmetrical molecule which as a high affinity for the active site of this molecule, thus blocking maturation of the virus (C2 inhibitor). The AIDS virus, HIV, has also been implicated in programmed cell death, apoptosis. L. Montagnier, the discoverer of the HIV virus, has suggested the use of anti- apoptotic agents along with antiviral agents for the prolongation of life in AIDS as well. These agents include: vitamin C, vitamin E, N-acetyl-cysteine, and superoxide dismutase. Retinoic acid, the active metabolite of Vitamin A has also been shown to inhibit apoptosis in T cells (see selected reading).
Gene therapy approaches to the inhibition the transcription of viral genes and the maturation of viruses are also being attempted. Several approaches are being attempted. What seems fruitful recently has been the targeting of antibodies to virus products to specific intracellular locations. So far, a modified antibody has been targeted to the endoplasmic reticulum (ER). The ER is the intracellular organelle which begins post-translational modification of proteins. An antibody directed against gp160 of the HIV was placed in a DNA vector. Transfection of cells with this DNA construct has been shown to inhibit maturation of the HIV virion. Other intracellular targets are being investigated for this virus (and other viruses), as well. Furthermore, other peptides and antibodies may be found by screening techniques such as those described in the drug discovery section.
Cardiovascular drugs have been under development by modern biotechnology companies. Transforming growth factor-▀s (TGF-▀s) (Genentech:TGF-▀1, Celtrix: TGF-▀2, and Oncogene Sciences:TGF- ▀3), are members of a family of over 18 proteins. TGF-▀s have pleotropic effects on many cell types including immune cells and fibroblasts. They are involved in immune memory, immune suppression and wound healing. TGF-▀s have been shown to mediate cardiac protection after ischemic injury. The protective effect is thought to be due to the inhibition of TNF (tumor necrosis factor) secretion by lymphocytes after ischemic injury. Antisense oligonucleotides (Genta) are some of the newer cardiovascular drugs which have shown promise. Specifically, antisense oligonucleotides for the c-myb oncogene have been shown to inhibit closure of the arteries after angioplasty.
Blood substitutes are being developed by Somatix. Artificial blood products is one of the major new biotechnology products of this company.
Wound healing is a complicated process involving the fibrobasts, epidermal cells, and the immune system. Future use of modern biotechnology will utilize cytokines to promote wound healing in people who cannot heal properly and to grow skin for transplants to severe disruptions in skin integrity, such as in burn patients. The TGF-▀s are involved in would healing (Genentech:TGF-▀1, Celtrix: TGF-▀2, and Oncogene Sciences:TGF-▀3). Oncogene Sciences, Inc. is developing the use of TGF-▀3 with Pfizer, Inc. and Ciba Geigy, Limited for the treatment of healing disorders such as oral mucositis which occurs after chemotherapy and wound healing in aged individuals.
Drug production..................................#
DRUG PRODUCTION:
Advances in biotechnology has spurred and demanded new methods of drug production. Bacteria have been used for years to produce recombinant proteins for pharmaceutical use. The most notable of these is insulin. Baculovirus systems have been recently developed and used for the production of drugs in vitro. Baculoviruses grow in insect cell lines which grow very rapidly in culture. These cells are used for the production of recombinant proteins from eukaryotic (non-bacteria) organisms. These cells are capable of post-translational modifications, like glycosylation. Therefore, the recombinant protein grown in these cells generally has a better bioactivity than those produced in bacteria. The most recent addition for methods of drug production developed by modern biotechnology are transgenic animals which produce pharmaceuticals in their milk. Goats, cows, and pigs are being developed which produce specific pharmaceuticals. Genepharming Europe BV, is developing cows that produce a human antimicrobial protein in their milk, lactoferrin. Pharmaceutical Proteins of Edinburg is developing transgenic sheep that will produce human alpha-1 antitrypsin in their milk. Transgenic goats are being developed by Genzyme Transgenics, a subsidiary of Genzyme, that will produce CFTR in their milk. CFTR is the defective protein in the disease cystic fibrosis. So far, only microgram quantities can be produced in these goats. They hope amounts suitable for clinical use can be produced within the decade. See the drug delivery section below. Transgenic plants are another method by which modern biotechnology products will be produced. See the agriculture section below.
Drug delivery....................................#Drug Delivery
Any compound conceived of or discovered by the above method requires a delivery system to be realized as a commercial bioactive molecule. A drug delivery system includes the method of application of the drug to the organism, the method of cell uptake, and stabilization of the compound for efficient delivery. This delivery can take the form of a virus, an antibody, a modified antibody, a chemical or a selected binding peptide. Seragen is developing fusion toxins which target activated T cells via the IL 2 receptor. So far, this treatment will be used for rhematoid arthritis, certain cancers (chemotherap resistant lymphomas) type I diabetes, and HIV infection. Furthermore, idiotype vaccines (killed MBP-reactive T cells are used to immunize against MS) have shown some promise and are in phase I trials. Derivatives of antibodies which are reduced in size are being developed for enhanced drug delivery (Creative Biomolecules) and Single Chain Antigen Binding (SCA(TM)) technology is being developed by Enzon. These smaller molecules are better able to penetrate tumor tissues. Viral vectors have already been used to treat disease and several studies will be undertaken very shortly. So far, human gene therapy has involved retroviral vectors only. For example, a crippled retrovirus with a the adenine deaminase gene inserted in it has been used for treatment of some cases of severe combined immunodeficiency (SCID). Future gene therapy will most likely include the use of other vectors such as herpes virus vectors and adeno-associated virus vectors as well as retroviruses. An adenovirus based gene therapy has been approved for use in cystic fibrosis. Studies are underway to deliver drugs by transfection of blood and cells with the gene therapy. PEGnology has been one of the most fruitful drug delivery systems of the last decade. Drugs are attached to polyethylene glycol and this gives them better stability and solubility properties. This has been used for IL 2, soluble CD4 (sCD4), and many other proteins. Enzon has an orphan drug patent on PEG- glucocerebrocidase for the treatment of Gauchers disease. Transdermal drug delivery is now being used to delivery of drugs through the skin by osmosis and electric currents. These "high- tech" systems are showing promise for the controlled continuous delivery of drugs.
Improvement of agricultural products.............#Improvement of Agricultural Products:
Plants are now in a "third wave of plant biotechnology". The "first wave" was to produce plants with improved agronomic traits, such as disease and herbicide resistance. Kirin Brewery of Japan has spent 2.5 million dollars for Calgene, a Campbell soup company, to develop disease resistant potato seedlings. Calgene has also developed BXN cotton which is resistant to the herbicide bromoxynil. Furthermore, attempts are being made to place antibody genes in tobacco plants at Scripps Research Institute in La Jolla, California as a new form of transgenic plant which produces its own pesticide. Antisense genes for plant viruses are being developed as well. The bean yellow mosaic virus (BYMV) antisense gene is hoped to protect ornamental flowers and legumes against the virus. Genetically modifying agricultural products to produce bacterial toxins is one approach to reducing the use of conventional insecticides. Bacillus thuringiensis delta toxins have been used to transform several crops and impart pest resistance to them. Plant genetic materials (PGMs) from plants resistant to disease are used to produce new plants that are resistant to disease. Ribozyme Pharmaceuticals is developing a corn which is more resistant to disease using their splicing technology. The "second wave" of plant biotechnology was to produce altered plants with better food processing traits, such as slowing down the ripening of tomatoes with antisense transgenes. This was accomplished by Calgene, who developed the "Flavr Savr tomato". Calgene has also developed a non-antisense or co-suppression recombinant DNA method to control tomato ripening that will be useful in many different plant species. This method of "Ethylene Control" utilizes a DNA construct for the introduction of an enzyme that controls ripening. A U.S. patent was granted to Calgene for this method of "Ethylene control" in July of 1993. Furthermore, a corn which produces more oil is being developed by Ribozyme Pharmaceuticals. The "third wave" of plant biotechnology is yielding plants that produce phamaceuticals, specialty chemicals, and biopolymers. This is being attempted by a number of companies, including Monsanto and Biosource Genetics Corp. Monsanto has transgenic tobacco plants that produce _-amylase and Michigan State University, in collaboration with Miescher Institute in Basel, Switzerland have produced turnips which produce human _-interferon. Future modifications of plants will include plants that use less fertilizer, improvement of nitrogen fixation or the creation of nitrogen fixing non-legume plants. Steps toward this goal have been the determination of the structure of enzymes involved in nitrogen fixation and the induction of nodules on non-nitrogen fixing plants which may one day hold microbes capable of fixing nitrogen, like legumes. Ornamental plants may undergo a transition induced by modern biotechnology. The luciferase gene, the gene that causes fire-flies to emit light, has been successfully transfected into plants. This may one day lead to a new generation of ornamental plants that emit light. So far, however, the
chemical substrate for luciferase.
Genetic engineering of livestock has been undertaken by a number of companies with limited success so far. Transgenic cattle with a leaner meat are being attempted. Genetic Engineering of Colorado has attempted to create cattle transgenic for the chicken myosin gene. This has not been successful, yet. Although the gene was inserted and the chicken gene was made, the animal developed muscle weakness early in life and was "put down" at a very young age.
Biological control...............................#BIOLOGICAL CONTROL
Biological control with techniques of modern biotechnology is being investigated for the maintenance of biodiversity, the control of pests, and prevention of the transmission of disease to humans. For example, recombinant vaccines have been developed to vaccinate wild animals against the rabies virus. Biological control of pests with infectious agents is being attempted for vermin of agricultural products. Parasitic wasps are being examined for the control of caterpillars which destroy a number of crops. Fungi which infect onion worm larvae are another example of an approach to biological control of agricultural products. Furthermore, bacterial toxins are being investigated for the control of insects as an alternative to insecticides (See the improvement of agricultural products section). Biological control of organisms in order to preserve biodiversity is now being scrutinized in Australia to preserve the numbats, an Australian marsupial. However, this approach has reasonably been questioned. Since this approach will sterilize red foxes and rabbits with viruses, the fear of the escape of these organisms from Australia and the sterilization of world populations of these two organisms is a reasonable concern.
Current interests of marine biotechnology include development of fish which are resistant to disease and with desirable growth characteristics, new treatments for fish diseases, and diagnosis of fish illnesses. Breeding fish with desired traits is being approached by classical genetic methods and by new technologies like the introduction of transgenes. The release of transgenic fish into the environment is now being considered by scientists and environmentalist groups. The national biotechnology center at the National Wildlife Federation is an environmental group that is considering this proposal. The Agricultural Biotechnology Research Advisory Committee (ABRAC) is developing safety standards. Treatment and diagnosis of fish diseases is an active interest of biotechnology, particularly for domestic production of fish. Fish ponds and holding pens are used for the farming of fish for human consumption. These environments are unnatural and lead to the rapid dissemination of disease. Therefore, methods of early detection and treatment are required for these environments. For example, Mirologix Biotech, Inc. is developing detection kits and treatments for: Bacterial kidney disease (Renibacterium salmonianrum), Furunculosis, (Aeromonas spp.), Vibrosis (Vibrio), and Red-mouth disease (Yersinia ruckeri).
Improvement of the environment...................#IMPROVEMENT OF THE ENVIRONMENT:
Improvements in the environment which can be the result of modern biotechnology will obviously include: clean water, photosynthesis, O2 production and C02 utilization, and removal toxic pollutants. Japan Research Development Corp. has committed $15 million over 5 years to Michigan State University for the development of microbes to improve the environment. Many other companies have been involved in this research as well. Previous successes have included genetically modified bacteria for digesting hydrocarbons.
Biodiversity and the maintenance and/or repair of the ecosystem are of current interests to the world and the biotechnology industry as well. There have been significant setbacks recently, however, and these are in part due to the biotechnology industry in the United States of America. The United States of America hesitated at the recent summit on the "Biodiversity Treaty" and it was not signed by the United States of America even though 98 countries adopted it. The Clinton administration has now signed it and it is waiting to be ratified by the senate. The United States of America was concerned that the treaty is merely an attempt to co-opt a technology that they already dominate, biotechnology. The explanation for their failure to sign the treaty is that its uncertain compulsory licensing scheme may defeat its own purpose, causing a greater destruction of the current biodiversity. However, there is now a controversy about whether to charge the West royalties for the use of PGMs (plant genetic materials) from developing countries. This demand has been increased recently in the wake of the biodiversity treaty. The maintenance of biodiversity and the world ecosystems is likely to require drastic measures which protect our biological resources and our biotechnology industries. The Biotechnology industry is likely to play a large role in maintaining biological diversity as well as creating a better living environment.
The identification of a new taxol source is an example of a recent success the biotechnology industry. Environmentalists have been concerned that the Pacific yew tree would be wiped out by the isolation of taxol from it. Now a new source has been identified, the fungus Taxomyces andreanae. Cytoclonal Pharmaceutical, Inc. has recently agreed to commercialize this form of taxol.
ECONOMICS OF THE BIOTECHNOLOGY INDUSTRY
The biotechnology industry has suffered serious setbacks in 1993. This drop in stock prices for the biotechnology industry has resulted from several blows to the industry: test results and the political climate. Furthermore, drug prices have come under a great deal of scrutiny and are also affected by the political climate as well as the structure of the health care centers. The move toward managed care networks has created large purchasers of health care products. These large purchasers are forcing companies to developed breakthrough and cost-effective products.
Test results have caused a major setback in the biotechnology industry recently. These have resulted from drugs targeting the septic shock that can result from infection of the blood stream after illness, trauma, or surgery. Centocor and Xoma corporation were both developing a version of monoclonal antibodies to endotoxin. Endotoxin is the component gram negative bacteria responsible for initiating septic shock. Centocor's "Centoxin-HA-1A" and Xoma's "E5" versions of these antibodieswere aimed at neutralizing the endotoxin. Both were deemed ineffective by the FDA on January 18, 1993 for treating septic shock. The second blow to drugs designed to treat sepsis came with the phase III clinical results of Synergen's drug, Antril. Synergen's stock price loss was dramatic. The price of Synergen's stock dropped 28 points, from 42.125 to 14.125 in one day, February 22, 1993. Antril is Synergen's preparation of an antagonist for interleukin 1 receptors called IL 1ra. Interleukin 1 is secreted by the immune system in response to components of bacteria and it initiates many of the symptoms of septic shock, such as hypotension. Administration of IL 1ra preparations, such as Antril, were to alleviate the effects of the IL 1 induced by the infections. Phase II clinical trials were very impressive for Antril, treated patients exhibited 27% mortality versus 64% for those who received the placebo. Phase III clinical trials were very poor, however, and this sent Synergen's stock down on February 22, 1993. Antril treated patients had a 29% mortality versus 34% for patients who received a placebo. A detailed chronology of Synergen's Antril failure can be found in appendix B. Soluble interleukin 1 receptor, developed by Immunex has also been deemed ineffective for septic shock. Xoma is in phase I clinical trials with another product aimed at treating septic shock, RBI-23. This compound is a recombinant version of human BPI. BPI, or bactercidal protein, also binds IL 1 and TNF. Therefore, it may have the effect of both inhibiting the action of the endotoxin induced IL 1 and TNF.
The political climate has also effected the prices of biotechnology stocks. There has been an uneasiness about the healthcare reforms that the Clinton administration will be initiating. The AMEX's biotechnology index, BTK, fell to 103 in August of 1993 from January's price of 170 and the CBOE's, BGX, stock index fell from 162 on December 31, 1992 to 101 in August of 1993. However, the Clinton administration allocated a 16% increase to the National Science Foundation (NSF) and a 3.3% increase to NIH. Together with postive fellings about the Clinton Administration's "Healthcare Reform", the BTK and the BGX are recovering in October of 1993 at 134 and 130, respectively.
Drug prices are also affected by the political climate. This in turn will naturally affect drug profitability and the stock prices of pharmaceutical and biotechnology firms. A recent report from the Office of Technology Assessment said that the market failed to control drug prices. Furthermore, drug firms have been accused of gouging the public. U.S. drug prices are higher in the U.S.A., and the U.S. Senate has accused the industry of price-gouging. They have claimed that the revenues of the pharmaceutical industry far exceed other industries and that market pressures do not affect the prices of drugs. Obviously, all of this affects drug prices. Despite these price criticisms, drugs have been shown to be the most cost-effective therapy and pharmaceutical firms invest more in research and development than any other industry, 16.7% or their revenues. Furthermore, the PMA claims there must be potential for high reward to justify the risk that pharmaceutical companies must take. Therefore, any controls imposed should not stifle drug development and will have to take this into consideration. The U.S. government has supported this as well. President Clinton's health care reform has imported no price controls for the pharmaceutical industry. An example of the biotechnology industry controlling its own prices has occurred for transgenic animals. There has been an uproar recently about the prices of transgenic mice and knock-out mice. GenPharm of Mountain view, CA was accused of charging 10 times more for "knockout" mice than non-profit organizations. The president of GenPharm told the National Academy of Science recently that researchers can buy the breeding pairs and then breed as many as they want for an annual fee of $1000.
INFORMATION ABOUT BIOTECHNOLOGY
Keeping up with the biotechnology industry: news and services. There are a number of investment newsletters available, several of which keep track of predominately biotechnology stocks (appendix C). For general stock analysis, "The Chartist", is highly rated. The AgBiotech stock letter, Medical Technology Stock Letter, and Sturza's Medical Investment Newsletter mainly focus on medical-technology companies. Investing often has interesting articles, but must be used with caution. For example, in January of 1993 claimed that Synergen stock "was nearing the end of a long fall from its all time high of 75". As mentioned above, Synergen's stock fell 28 points in one day to 14.25, with a high of 19 for the day. There are several databases that can be found on-line. Current Quotes and the Executive News Service of CompuServe Information Service (CIS) are very useful for tracking the daily activity of biotechnology stocks. Several folders can be made to track biotechnology news for inventions, results, and company reports. Furthermore, SmartScans and Knight-Ridder's Knowledge Index can be found on CIS. Knight-Ridder's Dialog, a more sophisticated database, has several files which are useful for obtaining specific information on biotechnology. See appendix B for a list of useful databases on Dialog. Mead Data Central of Dayton, Ohio, which offers Lexis and Nexis services, was to allow individual users access in 1991. GEnie, the computer service of General Electric also offers several useful databases similar to CIS. Furthermore, they offer a gate to the Dow Jones News/Retrieval service. The Prodigy service can be used for retrieving stock quotes.
INVESTING IN BIOTECHNOLOGY
Investing in the biotechnology industry can be very profitable. In the last couple of years there have been a large number of "winners" in the market. Immunex, Oncogene Sciences, Cellpro, and Biogen are all example of this. Immunex is now part of American Cyanamid after a merger with the Lederle Oncology Corporation in June of 1993. The recent fall of the stock price of several biotechnology companies, however, has introduced the possibility of loosing money by investing in biotechnology. For example, the fall of Synergen's stock by 70% in one day was an incredible unpredictable loss. Obviously, care must be taken when investing in biotechnology today. The establishment and maintenance of a successful personal portfolio of stocks of modern biotechnology companies will require intelligence and thought. This can be done by being informed of the goals of individual biotechnology companies and their approaches to developing and producing their biotechnology goals. Discount brokers can be found on most large computer services, such as Quick and Reilly and Spear Securities on CIS. Alternatively, one can invest in biotechnology investment funds. One fund which has done well is Global Health Sciences Fund of Denver, CO. Through much of the slump in biotechnology prices described above, they showed a $0.00/share loss. They can be reached at (800) 528-8765 or (303) 930-6521.
SELECTED READING:
Eisenberg, RS. 1992. Genes, patents, and product development. Science 257:903-918.
Marshall, E. 1991. The patent game: Raising the ante. Science 253:20-24.
Kiley, TD. 1992. Patents on random complementary DNA fragments?
Science 257:915-918.
Adler, RG. 1992. Genome Research: Fulfilling the public's expectations for knowledge and commercialization. Science 257 :908- 914.
Amato, I. 1992. Speeding up a chemical game of chance. Science 257: 1992.
Anderson, C. 1993. Clinton's technology policy emerges. Science 259: 1244.
Science. May 28, 1993. AIDS: The unanswered questions. Science 260:1254-1285.
Erickson, J. et al. 1990. Design, Activity, and 2.8┼ crystal structure of a C2 symmetric inhibitor complexed to HIV-1 protease. Science 249:527-534.
Vagelos, PR. 1991. Are perscription drug prices high. Science 252: 1080-1084.
Trends in Biotechnology
Saiki, RK, Gefland, DH, Stoffel, S, Scharf, SJ Higuchi, R, Horn, GT, Mullis, Erlich, HA. 1988. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487-494.
Koff, WC and Hoth, DF. 1988. Develoment and testing of AIDS vaccines. Science 241:426-432.
Geisow, MJ. 1993. Aching for approval, hoping for harmony - biotech product regulation. Trends in Biotechnology 10:107-110.
Liu, DT-Y. 1993. Glycoprotein pharmaceuticals:scientific and regulatory considerations, and the US Orphan Drug Act. Trends in Biotechnology 10:114-120.
Balter, M. 1991. How Europe regulates its genes. Science 252:1366-1368.
Friden, PM et al. 1993. Blood-Brain Barrier penentration in Vivo activity of and NGF conjugate. Science 259:373-377.
Rosen, DR et al. 1993. Mutations in Cu/Zn superoxide dismutase SELECTED READING (CONTINUED):
gene are associated with familial amyotrophic lateral sclerosis.
Gibbs, WW. 1993. Try, Try Again: Making antibodies more useful by making them more human. Scientific American 269:101-103.
Kohl, NE et al. 1993. Selective inhibition of ras-dependent transformation by a farnesyltransferase inhibitor. Science 260:1934-1937.
James, GL, 1993. Benzodiazepine peptidomimetics: Potent inhibitors of ras fanesylation in animal cells. Science 260:1937-1942.
Gougeon, M-L and L Montagnier. 1993. Apoptosis and AIDS. Science 260:1269-1270.
Ketajima, I. et al. 1992. Ablation of transplanted HTLV-I Tax- transformed tumors in mice by antisense inhibition of NF-KB. Science 258:1792-1796.
Nature 359:67. Antisense c-myb for athleroscerosis.
Hyde, SC. 1993. Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy. Nature 362:250.
Anderson, WF. 1992. Human gene therapy. Science 256:808-814.
Chatterjee, S et al. 1992. Dual-target inhibition of HIV-1 in Vitro by means of an adeno-associated virus antisense vector. Science 258:1485.
Culver, KW et al. 1992. In Vivo gene transfer with retroviral vector-producer cells for treatment of experimental brain tumors. Science 256:1550.
Trail, PA et al. 1993. Cure of Xenografted human carcinomas by BR96-Doxorubicin immunoconjugates. Science 261:212-215.
Rabizadeh, S, J Oh, L-t Zhong, J Yang, CM Bitler, LL Butcher, DE Bredesen. 1993. Induction of apoptosis by the low-affinity NGF receptor. Science 261:345.
Anderson, WF. 1992. Human Gene Therapy. Science 256:808-814.
Golumbek, PT, AJ Lazenby, HI Levitsky, LM Jaffee, H KArasuyama, M Baker, DM Pardoll. 1991. Treatment of established renal cancer by tumor cells engineered to secrete interleukin 4. Science 254:713.
SELECTED READING (CONTINUED):
Iwata, M, M Mukai, Y Nakal, and R Iseki. 1992. Retinoic acids
inhibit activation-induced apoptosis in T cell hybridomas and thymocytes. J. Immunol. 149:3302-3308.
Pietsch, T, U Kyas, U Steffens, E Yakisan, MR HAdam, W-D Ludwig, K Zsebo, and K Welte. 1992. Effects of human stem cell factor (c- kit ligand) on proliferation of myeloid leukemia cells: Hetererogeneity in response and synergy with other hematopoietic growth factors. Blood 80:1199-1206.
Betaseron Study. April 1993. Neurology.
Lehner, T, LA Bergmeier, C Panagiotidi, L Tao, R Brookes, LS Klavinskis, P Walker, J Walker, RG Ward, L Husaain, AJH Gearing, adn SE Adams. Induction of mucosal and systemic immunity to a recombinant simian immunodeficiency viral protein. Nature 258:1365-1369.
Mervis, J.. 1993. Regulations go Swimmingly. Science 261:542.
Stierle, A., G. Strobel, D. Stierle. 1993. Taxol and Taxane production by Taxomyes andreanae, endophytic fungus pacific yew.
Science 260:214.
Nijn, I, L das Neves, A van Kammen, H Franssen, and T Bisseling. 1993. Nod factors and nodulation in plants. Science 260:1764.
Palca, J. 1992. Testing target date looms, but will the vaccines be ready. Science 257:1472.
Travis, J. Putting antibodies to work inside cells. Science 261:1113.
McGaughey, W.H. and M.E. Whalon. 1992. Managing insect resistance to Bacillus thuringiensis toxins. Science 258:1451.
Morell, V. 1993. Australian pest control by virus causes concern. Science 261:683.
Weiner, H.L., G. A Mackin, M. Matsui, E.J. Orav, S.J. Khoury, D.M. Dawson, D.A. Hafler. 1993. Double-blind pilot trial of oral tolerization with myelin antigens in multiple sclerosis. Science 259:1321.
Zhang, J., R. Medaer, P. Stinissen, D. Hafler, and J. Raus. 1993. MHC-restricted depletion of human myelin basic protein-reactive T cells by T cell vaccination. Science 261:1451.
Hsieh, C.-S., SE Macatonia, CS Tripp, SF Wolf, A O'Garra, KM Murphy. 1993. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science 260:547-549.
Cohen, J. T cell shift: Key to AIDS therapy? Science 262:175- 176.
APPENDIX A
The Overlapping Stocks of the CBOE and the AMEX are:
Amgen (AMGN)
Biogen (BGEN)
Centocor, Inc. (CNTO)
Chiron (CHIR)
Cytogen Corp. (CYTO)
Enzon, Inc. (ENZN)
Gensia Phamaceuticals (GNSA)
Genzyme Corp. (GENZ)
Greenwich Pharmaceuticals Inc. (GRPI)
*Immunex (IMNX)
Liposome Co. Inc. (LIPO)
Synergen (SYGN)
Xoma (XOMA)
*Immunex has been replaced by Immune Response Corporation on the AMEX due to a merger with American Cyanamid.
The AMEX's Biotechnology Index
BTK: Total Capitalization as of June 2, 1993 was 91.8 million.
Amgen (AMGN)
Genzyme Corporation (GENZ)
Biogen (BGEN)
Immune Response Corporation (IMNR)
Synergen, Inc. (SYGN)
Enzon, Inc. (ENZN)
Xoma Corporation (XOMA)
Gensia Pharmaceuticals, Inc. (GNSA)
Cambridge Biotech Corp.
Liposome Company, Inc. (LIPO)
Centocor, Inc. (CNTO)
Chiron Corp. (CHIR)
Scios Nova, Inc. (SCIO)
Cytogen Corp. (CYTO)
Greenwich Pharmaceuticals, Inc. (GRPI)
APPENDIX B
The following are quotes from the Denver Post concerning the fall of Synergen's stock price:
January 19, 1993
BIOTECH STOCKS WOBBLY:Synergen bounces on Centocor news.
January 28, 1993
HOAX POUNDS BIOTECH STOCKS:Synergen shares plummet after false drug-test report.
"One person's hoax, based in impatience or perhaps greed, has sent the entire biotech industry reeling and the stock of Boulder-based Synergen Inc. plunging"
"...on Tuesday, someone identifying himself as a doctor involved n Synergen's study phoned in phony results to a medical newsletter. Calling himself a physician from UCLA, he said the drug known a Antril simply didn't work."
"While the report was'nt true, it was enough to sent Synergen's stock dropping more than $6 per share."
""Unfortunately, rumors of this sort no matter how specious they may be are impossible to refute definitively, especially when they're in such a vulnerable period"......."
""These rumors are illogical and spurious. The greatest probability remains that the results (of Antril tests) will be very good.""
"The mess started Tuesday (1/26) when Evan Sturza of
Sturza's Medical Investment Letter received a call from a "Dr. Miller", who claimed to have participated in the University of California at Los Angeles arm of the study of Antril, Synergen's sepsis drug.""
February 18, 1993
SYNERGEN NET LOSS SURGES
"Synergen Inc., a Boulder-based biopharmaceutical company, reported more than a 1,000 percent increase in net loss for the fourth quarter ....."
February 22, 1993
SYNERGEN STOCK PLUNGES 70%:Weak drug price sinks NASDAQ
APPENDIX C
DIALOG DATABASES FOR BIOTECHNOLOGY INFORMATION
BIOBUSINESS: Covers current and retrospective biotechnology information for business professionals, File number 285, 1985- present.
BIOCOMMERCE ABS AND DIRECTORY: Abstract records and company profiles of the biotechnology business, File number 286, 1981- present.
BIOTECHNOLOGY ABSTRACTS: Coverage of biotechnology publications, File 357, 1982-present
CLAIMS/REASSIGNMENT: Patents that have been reviewed by the government and have been reassigned, File number 123.
DERWENT BIOTECHNOLOGY ABSTRACTS: File numbers 350 and 351.
DRUG INFORMATION FULLTEXT (DIF):File number 229. Brief descriptions of all new drugs approved by the FDA. It corresponds to AHFS drug information and the Handbook on Injectable drugs publications.
EMBASE: File numbers 72 (1974-present), 73 (1985-present), 272 (selected records from 8 weeks of 1989).
INVESTEXT: Information about companies, industry, geographic reports etc., File numbers 545 (1982-present), 277 (1988- present).
IMSWORLD R&D FOCUS: Latest Scientific and commercial developments in international pharmaceuticals, File number 445.
JAPIO: Japanese patents, File number 347
PATENTS: Full text of patents, File number 654.
PATFUL: United States of America Patents, three files: 654 (1990), 653 (1980-1989) and 652 (1971-1979).
PHIND: Pharmaceutical and healthcare industry news database, agriculture is covered as well, AGROW and Animal Pharm., File number 129.
PREDICASTS: Newsletters classified by the industry, it includes biotechnology, File numbers: 16 (1972-present), 216 (1988-1990), 636 (1988-present).
UNLISTED DRUGS: File Number 140.
APPENDIX C (CONTINUED):
CompuServe Databases
DISCLOSURE
VALUE-LINE
GEnie Databases
VESTOR
Dow Jones/News Retrieval
INVESTMENTS IN BIOTECHNOLOGY IS SHAREWARE:
The author has a Ph.D. in Microbiology/Immunology and expects a $30.00 fee for use of this material. Its last update was October 15, 1993. Register your copy by sending $30.00 to P.O. Box 18978, Denver, CO 80218-0978. The institutional and corporate rate for 10 or more copies is $20.00/copy. Copyright, 1993. It is planned that this file will be updated frequently. There is no charge for updates of the same whole number. Updates will carry a $15.00 charge. The author can be reached on CIS at 71240,2762.
REGISTRATION OF INVESTMENTS IN BIOTECHNOLOGY Rel 1.63
REGISTRATION : Investments in biotechnology is being distributed on a SHAREWARE principle. Each individual user is responsible for the registration of their copy if they decide to keep and use it.
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