Large Scale Solid Phase Organic Synthesis



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Company Profile Large Scale Solid Phase Organic Synthesis Enabling the chemistry of your strategy and aspirations www.cblbiopharma.com www.cblpatras.gr. Europe & Middle East CBL-Patras GR-26000, Patras Tel: +30 2610 647 339 North America & Japan CBL Biopharma LLC PO Box 11069 Boulder, Colorado 80301 Tel: 303-317-3516 Email: service@cblbiopharma.com Web: www.cblbiopharma.com 1

Enabling the chemistry of your strategy and aspirations. Transforming your projects into success. CBL (Chemical and Biopharmaceutical Laboratries in Patras) was founded in 1990 by Prof. Dr. Kleomenis Barlos, to provide industry with novel chemistries in key areas of solid phase peptide and organic synthesis. CBL Biopharma was established in the United States to expand the organization s presence onto two continents. With the expansion of a state-of-the-art facility, CBL has increased its capacity to produce cgmp material from the grams to 500 kilogram scale. This investment reaffirms not only its commitment to quality and guideline compliance, but also to meet the needs of the marketplace. Being a primary manufacturer and developer of innovative reagents for peptide synthesis, CBL carries over 400 individual compounds for solid-phase organic and peptide synthesis. CBL operates out of 6,000 square meter laboratory and production space divided on three buildings spanning over 25,000 square meter land. CBL has become the ideal source as your industrial and commercial-scale manufacturer of proprietary and generic GMP and non-gmp peptides. CBL takes great pride in the partnerships formed from the beginning to completion of scientific and manufacturing endeavors with its clients. Solid phase organic synthesis is nowadays the core technology employed in the automated routine peptide synthesis as well as in combinatorial chemistry. The launch by Roche of the anti-hiv treatment Fuzeon, a chemically complex peptide, marked a paradigm shift in the pharmaceutical industry. Your source for non-gmp and GMP peptides. Solid phase peptide synthesis batches up to 500 kg. CBL s strong partnership with Roche to provide key materials and technology for the launch of Fuzeon, a first-in-class anti-hiv pharmaceutical, provided CBL with the recognition for its outstanding performance and commitment in reducing time-lead, cost and risk. CBL Patras was awarded the title of Roche Supplier of the Year for 2001 and 2002. Large scale solid-phase synthesis is the specialty of CBL. Development of a highly automated, efficient plant has enabled CBL to readily scale-up solid and solution phase synthesis. With these technological achievements, CBL stands as a global leader in solid-phase peptide and organic synthesis. Even with the most complex peptides, CBL s scientific expertise will join your company in meeting your goal. From the gram to kilogram scale, CBL works to provide innovative solutions and support to its partners. 1 2

Peptide API production at the leading edge. Cost effective, time efficient. As a primary manufacturer, CBL has always been on the leading edge of developments in peptide synthesis. CBL is specialized in the production of pharmaceutical grade peptides. With three large scale reactors and equipment for medium and small scale production completely dedicated to solid-phase peptide synthesis, CBL holds a unique state of the art manufacturing infrastructure. The composition and distinct capabilities at CBL shortens production time from the gram to ton scale. Designed to meet your needs from gram to commercial production. All final products and starting materials are available at industrial scale and are produced in-house to cgmp standards. CBL - where expectations are met Laboratory and Pilot plant Lab to development scale uses glass lined reactors and filters: Scale gram to multi-kilogram Peptide Synthesizer Small scale: Applied Biosystems 433 A Medium scale: Prelude Large scale: PSI Model 500/500E Solution & Solid Phase Reactors: 1 x 17 litres 1 x 100 litres 2 x 250 litres 4 x 200 litres glass-lined 2 x 80 litres glass-lined 1 x 20 litres glass-lined Chromatography Waters Preparative HPLC 4000 Medium scale Knauer Preparative HPLC with Merck column SP 100 Freeze Dryer (Lyophilizer) Virtis Lyophilizer: Genesis 35 SQ Super XL Manufacturing plant Manufacturing operates in glass lined reactors and filters, batch sizes up to 500 kg in solid phase peptide synthesis are possible. Fully automated solvent recycling takes place in five different distillation plants, columns up to 60 theoretical plates, capacities up to 2 m³ per hour. Scale kilogram to 500 kilogram Solution & Solid Phase Reactors: 2 x 1600 litres glass-lined 3 x 2500 litres glass-lined 2 x 4000 litres glass-lined 1 x 2500 litres stirred/pressure filter/dryer, stainless steel, 1 x 2500 litres stirred/pressure 1 x 1000 litres stirred/pressure 1 x 600 litres stirred/pressure 1 x 250 litres stirred/pressure 1 x 60 litres stirred/pressure 2 x 2500 litres pressure filter, glass-lined. 3 x 1600 litres pressure filter, glass-lined. Chromatography Novasep s Preparative HPLC unit: HyperSep LSC and LC 450. Preparative HPLC unit equipped with Procrom 450 mm internal diameter column, 180 stainless steel slurry tank, solvent module with maximum flow rate 200 litres/h, automatic module. Freeze dryer (Lyophilizer): 2 x GEA S Lyophil: Lyovac with an effective charging surface approximately 0.6 m 2 and 10 m 2 respectively. The manufacturing process is fully automated. Control equipment is working under the highest standards and conforms to cgmp. 4

Technologies Solid-phase synthesis CBL applies mainly Fmoc strategies to produce gram to ton quantities of peptides by solid-phase synthesis in reactors up to 4000 litres. For example, a peptide with a lot size of 200 kilogram, produced with solid phase technologies under cgmp conditions, generally is delivered with 2-4 months. Solution-phase synthesis CBL manufactures peptides by solution-phase technology. This is not limited to the production of peptides. For example, in the case that a key intermediate is a complex organic molecule, CBL facilities accommodate these conditions. Purification CBL employs a variety of purification techniques, including HPLC. Service Contract manufacturing At CBL, manufacturing is focused on the production of large volumes of pharmaceutical grade peptide APIs. A combination of many years of experience and expertise in solid-phase peptide and organic synthesis shortens project time considerably, whether in synthetic development, scale-up to the ton range, or process optimization. This is further supported by the constant stream of new and innovative reagents originating at CBL. When it comes to outsourcing peptide manufacturing, CBL will partner with your company to provide economical and time efficient services. Our services include: c-gmp production of gram to ton batches Scale-up studies in kilolabs and pilot plant Custom R&D, process chemistry research Process development Process validation Development and validation of analytical methods 5

Quality management More than 30% of CBL s workforce is directly involved in quality through the Quality Control and Business & Process Documentation teams. As a result, regulatory compliance and targeted customer specifications are close to fail-safe for every project that CBL embarks upon with its clients. All products are available in industrial scale and are produced in-house to cgmp standards. Each product is supplied with a lot specific Certificate of Analysis. Client tailored specifications are available upon request. HPLC Systems LC-MS analytical: Agilent 1100 Series LC/MSD Analytical HPLC : Water s HPLC system Analytical LC : Water s 2695 UPLC-MS(QTOF): Water s QTOF MICRO Proteomic High Flow system HPLC-derivatization: Agilent 1100 Series HPLC with PCx500 GC Systems GC: 2 x Agilent 6890 N GC GC-MS: Agilent 5973 N Other Optical rotation,: Perkin Elmer Instrument, model 341 polarimeter Particle size distribution Malder, mastersize 2000. Photometer: Spectroquant Nova 60 Colorimeter: Hunter Lab s Color Quest XE Microscope: Leica M 26 Steromicroscope Karl-Fischer Titration: Schott s Titroline KF Melting Point: Electrothermal 910 CBL is committed to meet and exceed customer s expectations by providing them with dependable, reliable products and services. The environment created by CBL for its customers, suppliers and employees cultivates and stimulates continuous improvement aimed toward Total Customer Satisfaction 7 8

Product range Fmoc amino acids Resins for Solid Phase Peptide and Organic Synthesis FAA-1005 Fmoc-L-Ala-OH BR-1000 Aminomethyl polystyrene FAA-1010 Fmoc-L-Arg(Pbf)-OH BR-1005 Aminomethyl polystyrene Our comprehensive product range includes more than 400 products in the area of: Resins for Solid Phase Peptide and Organic Synthesis Amino acid 2-chlorotrityl resins Resin-bound diamines Resin-bound aminothiols Resin-bound aminoalcohols Mercaptoacids bound through their thiol function onto 2-chlorotrityl resin Mercaptoacids bound through their thiol function onto 4-methoxytrityl resin N-α-Fmoc amino acids loaded to 4-methylbenzhydryl resin N-α-Fmoc-amino acids Amino acid derived diamines N-α-Trt-amino acids Homoserine derivatives Pegylation reagents All products are available in multi-kilogram to ton range. Standard items are available form stock in up to 200 kilogram lots. Our quality is of the highest standard and conforms to cgmp. To receive a copy of our latest catalogue please visit www.cblpatras.gr or call us today. Please see our website, for a complete list of our products at www.cblbiopharma.com or www.cblpatras.gr. FAA-1015 Fmoc-L-Asn(Trt)-OH FAA-1020 Fmoc-L-Asp(tBu)-OH FAA-1040 Fmoc-L-Cys(Trt)-OH FAA-1043 Fmoc-L-Gln(Trt)-OH FAA-1045 Fmoc-L-Glu(tBu)-OH H2O FAA-1050 Fmoc-Gly-OH FAA-1090 Fmoc-L-His(Trt)-OH FAA-1110 Fmoc-L-Ile-OH FAA-1120 Fmoc-L-Leu-OH FAA-1125 Fmoc-L-Lys(Boc)-OH FAA-1150 Fmoc-L-Met-OH FAA-1175 Fmoc-L-Phe-OH FAA-1185 Fmoc-L-Pro-OH FAA-1190 Fmoc-L-Ser(tBu)-OH FAA-1210 Fmoc-L-Thr(tBu)-OH FAA-1225 Fmoc-L-Trp(Boc)-OH FAA-1230 Fmoc-L-Tyr(tBu)-OH FAA-1245 Fmoc-L-Val-OH Preloaded Resins Preloaded 2-chlorotrityl resins Fmoc-amino acids attached to Rink amide 4-methyl benzhydrylamine resin N-α-Fmoc amino acids attached to 4-Methoxybenz hydryl resin N-α-Fmoc amino acids attached to 4-Methylbenzhydryl resin Mercaptoacids bound through their thiol function onto 4-methoxytrityl resin Mercaptoacids attached to 4-chlorotrityl resin Resin-bound Amino alcohols Resin-bound Diamines Resin-bound Aminothiols N-(2-Chlorotrityl resin) amino acids BR-1180 BR-1185 BR-1060 BR-1065 BR-1330 BR-1340 BR-1310 BR-1315 BR-1300 BR-1305 Innovations from CBL New PEG derivatives Bromo-MBH polystyrene resin (100-200 mesh, 1.2-2.2 mmol/g) Bromo-MBH polystyrene resin (200-400 mesh, 1.2-2.2 mmol/g) 2-Chlorotrityl chloride resin (100-200 mesh, 1.0-1.6 mmol/g) 2-Chlorotrityl chloride resin (200-400 mesh, 1.0-1.6 mmol/g) Rink amide AM resin (100-200 mesh, 0.4-0.9 mmol/g) Rink amide AM resin (200-400 mesh, 1.0-0.9 mmol/g) Rink amide 2-chlorotrityl resin (100-200 mesh, > 0.4 mmol/g) Rink amide 2-chlorotrityl resin (200-400 mesh, > 0.4 mmol/g) Rink amide MBHA resin (100-200 mesh, 0.4-0.9 mmol/g) Rink amide MBHA resin (100-200 mesh, 0.4-0.9 mmol/g) Altering the properties, for example bio-availability or solubility profile, of various biologically active compounds can be done via pegylation or the attachment of polyamides. It has been shown that polyamides are neither toxic nor immunogenic [1]. The pegylation method suffers one major drawback; it is very difficult to synthesize PEGchains with a precise length (defined number of monomeric units). Therefore usually a mixture of PEG-chains with different lengths is applied to modify the target molecule. As a result the identical solvation effects on, for example the surface of a protein can not be guaranteed, when it comes to repeating same pegylation procedure, but this is mandatory when drug development is the task. Based on the work of Keith Rose [2] and Gerd G. Kochendoerfer [3] CBL has developed a method which allows to synthesise polyamid-peg-conjugates with precise molecular weight at any length desired. The viability of using polyamides with small PEG-chains to modify the biological properties has been shown in the case of haemopoietin [2,3]. In this case the enzyme resistance had been enhanced. 1. Hai, et al., Bioconj. Chem. 9:645-654 (1998) 2. Rose, Keith; United States Patent Application 20030228274 3. Gerd G. Kochendoerfer et all., Science 7 February 2003 Vol. 299. no. 5608, pp. 884-887 CBL is pleased to announce that polyamide-peg-chains are now available at industrial scale. PEG-1000 N-Fmoc-N -succinyl-1,12-diamino-4,7,10-trioxa-triadecane PEG-1010 N-Mmt-N -succinyl-1,13-diamino-4,7,10-trioxa-triadecane PEG-1030 PEG-1050 PEG-1160 PEG-1100 N-Fmoc-N -succinyl-1,12-diamino-4,7,10-trioxa-triadecane 2-chlorotrityl resin N-Fmoc-N -succinyl-1,12-diamino-4,7,10-trioxa-triadecane2-benzhydryl resin 1,12-diamino-4,7,10-trioxa- triadecane trityl resin N -succinyl-1,12-diamino-4,7,10-trioxa-triadecane 9 10

Contacts @ CBL CBL Patras US Office Bulk inquiries API & Custom manufacturing GR-26000, Patras GREECE Tel: +30 2610 647 600 CBL Biopharma LLC PO Box 11069 Boulder, Colorado 80301 Tel: 303-317-3516 Email: service@cblbiopharma.com GR-26000, Patras GREECE Tel: +30 2610 647 338 GR-26000, Patras GREECE Tel: +30 2610 647 338