Schering-Plough

The Schering-Plough Corporation (SP) is a research-based company engaged in the discovery, development, manufacturing and marketing of pharmaceutical products worldwide. SP is also one of the ten largest US based pharmaceutical companies and in 2001 its annual revenue was about $ 9,800 million, of which, more than 10% was invested in R&D activities ($ 1,200 million). Schering Plough's R&D operations are managed by the scientific arm of the Corporation: the Schering-Plough Research Institute (SPRI).

SPRI is located worldwide and it is involved in seven major therapeutic areas: allergy, immunology, infectious diseases, oncology, central nervous system, cardiovascular and gene therapy. The headquarters are in Kenilworth, New Jersey. All of the therapeutic areas are covered at this site and professional expertise range from initial Drug Discovery to full clinical development. There are two additional New Jersey sites which are closely linked to Kenilworth: one is in Madison (Pharmacy) and the other in Lafayette (Pre-clinical development). There are also additional US sites in California, one is in Palo Alto near Stanford University (DNAX, focused on immunology, new protein discovery, cell signalling and gene targeting technology) and one in San Diego (Canji focused on gene therapy). European SPRI sites are in France (Dardilly, near Lyon) specialised in immunology research, Switzerland (Schachen, near Luzern) devoted to chemical and biotechnological development and Milan focused on Neuroscience.

In addition to the above, extensive support to the internal research programs is given by strategic alliances with external centres of excellence. These include Human Genome Sciences, Genome Therapeutics, Incyte, Myriad Genetics, Transgène, Deltagene to name but a few.

The Italian Research Centre and its organisation

The Italian unit of the SPRI is located within the San Raffaele Biomedical Science Park in Milan. The centre has a long-standing reputation in neuroscience research as supported by publications and patents in the area. In the past, activity was spread over different disease areas such as Parkinson's, Alzheimer's, stroke and neuropathic pain but in 2001, a significant operational reorganisation was undertaken in order to optimise the chance of success.

The first strategic change was to take responsibility of projects in the neuropathic pain area only. This means that all our efforts are focused on a single disease, with the aim of reaching a critical mass. Three technology platforms have been created to develop the technologies necessary to support the activities from target identification/validation, through hit identification, and lead optimisation as required by the new assignment. The technology platforms were identified based on areas of expertise and the various phases of the process. These include Neurobiology, Molecular Cell Biology and Neuropharmacology sections respectively.

In the new organisation the Neurobiology section was designed to support the early phases of target identification and validation. This role was covered by developing expertise in the analysis of gene and protein expression. Gene expression is studied using two complementary techniques, real time PCR that allows a quantitative analysis of expression and in-situ hybridization which permits transcripts to be localized to the single cell level. Biochemical techniques such as FACS, immunocytochemistry, histological staining and western blotting are also routinely used to analyse the pattern of distribution and the expression level of proteins.
The Molecular and Cell biology section is charged with supporting the later stages of target validation, the hit/lead identification and the lead optimisation phases in-vitro. This has been achieved by developing cutting edge technologies to clone and transfect target genes into appropriate cell systems (to obtain transient or stable cell lines for functional and/or screening studies) and by gaining expertise in the development of relevant cell based assays. With regards to cell based assays, a major advancement has been the acquisition of state-of-the-art instrumentation known as Flipr (Fluorometric Imaging Plate Reader). This automated equipment can test up to 10,000 samples a day thus allowing a medium to high throughput capability to run, full screening programs for new chemical entities entirely in house.
The role of the Neuropharmacology section is to support the target validation and the lead optimisation phases in-vivo. This is being achieved by developing expertise in the identification, setting-up and running of the relevant disease animal models to investigate the pathophysiogy of neuropathic pain as well as to test new chemical entities in-vivo.

Finally, a key feature of the re-organisation of the Italian research centre has been the full integration into the Schering-Plough drug discovery strategy and process. To this aim interactions with SPRI headquarters are implemented on a regular basis by teleconferences, videoconferences and frequent exchanges of researchers between facilities.

Therapeutic area of interest: neuropathic pain

The Italian unit has been given the lead of neuropathic pain research. In this disease current medications are not always fully effective and since none of them has been specifically designed for chronic pain, their use is often associated with significant side effects. Therefore, the identification of relevant medicines for neuropathic pain is a significant opportunity both scientifically and commercially.
Pain is a subjective and multidimensional experience comprising several components: sensory (e.g. intensity, duration, location), affective (e.g. unpleasantness, emotional, motivational) and cognitive (e.g. awareness of the implications, fear, anxiety). In most cases, the sensation of pain is produced either by stimuli which are intense enough (noxious) to cause tissue damage, or by nerve injury. The perception of pain can be influenced by many different factors that can be, by themselves, major determinants of pain, e.g. psychological factors. A formal definition of pain has been proposed by the International Association for the Study of Pain subcommittee on taxonomy that describes pain as "a highly subjective, unpleasant sensory and emotional experience associated with actual or potential tissue damage or described in terms of such damage". Chronic pain is elicited by peripheral damage due to injury or to a disease and it has more serious consequences than acute pain since damage overwhelms the body's repair capabilities, as it is often associated with the disruption of nerve endings. Chronic pain is a consequence of several clinical disorders, it afflicts a large proportion of the population and despite the remarkable advancement in understanding the disease it still remains an area of highly unmet medical need.

The commitment to excellence

SPRI firmly believes that a successful research group always needs to be at the front edge of the discovery effort and has to keep updated with technological evolution. To reach this objective the confrontation with the external research environment is essential and it takes place through participation at international scientific meetings, collaboration with groups in academic institutions and the organisation of seminars with leading scientists. In order to enrich the research environment in the Milan group, foreign scientists are encouraged to join the centre. One of the opportunities offered to foreign scientists is the possibility to undertake post-doctoral fellowships. These are granted to our centre by the European community to help recruit young and talented researchers from other European countries.
As in any company the experimental work is performed under strict confidentiality rules. This is to prevent the possible premature disclosure of information that could damage the competitive position of Schering-Plough. However, publication of experimental results obtained during the research effort is an important opportunity to communicate to and interact with the scientific community as well as to establish a solid, worldwide reputation in the disease areas of interest. These aspects are fundamental to the development of a successful research program not only in the academic environment but also in the Pharmaceutical industry. Therefore, publication of experimental results is encouraged to assure that the group has the necessary visibility and integration into the international scientific community. The fact that our research group has published more than 40 articles in the last five years is a testament to this ethos. We are however are also acutely aware of the commercial implications of our research and to this end have contributed significantly to the company patent portfolio by filing several patents.

List of selected publication of the last three years

Campanella M, Sciorati C, Tarozzo G, and Beltramo M Flow cytometry analysis of inflammatory cells in the ischemic rat brain. Stroke (2002) 33:586-592.

Tarozzo G, Campanella M, Ghiani M, Bulfone A, and Beltramo M Expression of fractalkine and its receptor, CX3CR1, in response to ischaemia-reperfusion brain injury in the rat. Eur. J. Neurosci (2002) 15:1-7.

Chishti MA, Yang DS, Janus C, Phinney AL, Horne P, Pearson J, Strome R, Zuker N, Loukides J, French J, Turner S, Lozza G, Grilli M, Kunicki S, Morissette C, Paquette J, Gervais F, Bergeron C, Fraser PE, Carlson GA, George-Hyslop PS, and Westway D Early-onset amyloid deposition and cognitive deficits in transgenic mice expressing a double mutant form of amyloid precursor protein 695. J Biol Chem. (2001) 276(24):21562-70.

Corradini L, Briscini L, Ongini E, and Bertorelli R The putative OP(4) antagonist, [Nphe(1)]nociceptin(1-13)NH(2), prevents the effects of nociceptin in neuropathic rats. Brain Res. (2001) 905:127-33.

El Yacoubi M, Ledent C, Parmentier M, Bertorelli R, Ongini E, Costentin J, and Vaugeois JM Adenosine A2A receptor antagonists are potential antidepressants: evidence based on pharmacology and A2A receptor knockout mice. Br J Pharmacol. (2001) 134:68-77.

Grundy RI, Rabuffetti M, and Beltramo M Cannabinoids and neuroprotection. Mol. Neurobiol. (2001) 24:(1-3):29-52.

Bertorelli R, Calo G, Ongini E, and Regoli D Nociceptin/orpanin FQ and its receptor: a potential target for drug discovery. Trends Pharmacol Sci. (2000) 21:233-4.

Citterio F, Corradini L, Smith RD, and Bertorelli R Nociceptin attenuates opioid and gamma-aminobutyric acid (B) receptor-mediated analgesia in the mouse tail-flick assay. Neurosci Lett. (2000) 292:83-6.

Grilli M, Diodato E, Lozza G, Brusa R, Casarini M, Uberti D, Rozmahel R, Westaway D, St George-Hyslop P, Memo M, and Ongini E. Presenilin-1 regulates the neuronal threshold to excitotoxicity both physiologically and pathologically. Proc Natl Acad Sci U S A. 2000 97(23):12822-7.

Rabuffetti M, Sciorati C, Tarozzo G, Clementi E, Manfredi AA, and Beltramo M Inhibition of caspase-1-like activity by Ac-Tyr-Val-Ala-Asp-Chloromethyl ketone induces long-lasting neuroprotection in cerebral ischemia through apoptosis reduction and decrease of proinflammatory cytokines. J. Neurosci. (2000) 12:4398-4404.

Grilli M, Barbieri I, Basudev H, Brusa R, Casati C, Lozza G, and Ongini E Interleukin-10 modulates neuronal threshold of vulnerability to ischaemic damage. Eur J Neurosci. (2000)12(7):2265-72.

Grilli M, and Memo M Possible role of NF-kappaB and p53 in the glutamate-induced pro-apoptotic neuronal pathway. Cell Death Differ. (1999) 6(1):22-7.

Bertorelli R, Corradini L, Rafiq K, Tupper J, Calo G, and Ongini E Nociceptin and the ORL-1 ligand [Phe1psi (CH2-NH)Gly2]nociceptin(1-13)NH2 exert anti-opioid effects in the Freund's adjuvant-induced arthritic rat model of chronic pain. Br J Pharmacol. (1999)128:1252-8.

www.sch-plough.com