XL-protein and Wacker demonstrate high-yield production of PASylated biopharmaceuticals using ESETEC®

Munich / Jena, June 11, 2012 – Wacker Biotech and XL-protein have successfully completed a feasibility study on the produc­tion of PASylated therapeutic proteins using WACKER’s ESETEC® E. coli secretion technology. In the study, WACKER technology was used to investigate the production of a PASylat­ed human growth hormone as model protein. WACKER devel­oped a cell line which, utilizing the ESETEC® technology, generated high yields of the PASylated protein. The PASylation® technology enables the development of biopharmaceuticals with extended plasma half-life, thus leading to a new generation of drugs with improved tolerability and requiring less frequent administration due to their longer lasting activity.

The aim of the study was to assess whether clinically established therapeutic proteins that have been improved by PASylation® can be produced in high yields by using state-of-the-art, cost-effective manufacturing processes. The study used WACKER’s proprietary E. coli-based secretion technology to produce a PASylated human growth hormone.

The feasibility study demonstrated that the desired PASylated protein, carrying a 600 amino-acid sequence consisting of proline, alanine and serine, can be successfully produced with ESETEC®. In addition, it also led to an efficient cell line which, during fermentation, secretes the correctly folded, fully functional protein into the culture medium in high yield (3-4 g/l). This is remarkable, as the availability of sufficient quantities of amino acid building blocks from the bacterial primary metabolism for the biosynthesis of large PASylated proteins represented a potential challenge. Moreover, detailed analytical studies showed that the PASylated human growth hormone was produced with high homogeneity in monomeric, non-aggregated form, retaining a remarkably high affinity for its receptor.

Compared to PEGylation, the current standard technology for extend­ing the plasma half-life of biopharmaceuticals, PASylation® has the major advantage of avoiding an expensive and poorly selective chemical modification step during production. Moreover, unlike the chemical polymer PEG, the PAS amino acid sequences are biodegradable and therefore do not accumulate in cells or organs.

ESETEC®, WACKER’s patented E. coli-based secretion system, is a well-established technology for producing proteins and antibody fragments cost-efficiently. It is based on a patented strain of E. coli K12, which is capable of secreting recombinant proteins in their native conformation into the culture broth during fermentation. Secretion into the medium facilitates the purification of the target protein considerably, since complicated process steps such as homogenization and refold­ing are no longer necessary, making the entire manufacturing process significantly more efficient and cost-effective. A number of biopharmaceuticals (biologics) manufactured with ESETEC® are already being evaluated in preclinical and clinical trials.

According to Dr. Thomas Maier, managing director of Wacker Biotech GmbH, he and his team “are delighted by the early results of our col­laboration with XL-protein. PASylation® is an extremely interesting technology for selectively improving the properties of biopharmaceu­ticals. With our innovative ESETEC® secretion technology, such prod­ucts can now be manufactured cost-effectively on an industrial scale.

Prof. Dr. Arne Skerra, CEO of XL-protein GmbH, concurs. “We are extremely satisfied with the results of the feasibility study. This allows us to bypass the expensive in-vitro modification steps as they are required in other processes currently available. The successful combination of our PASylation® technology with WACKER’s ESETEC® secretion technology has a high potential for synergies with respect to the efficient and rapid production of novel therapeutic proteins using an established host organism like E. coli with its well-known advantages.”

XL-protein announces in vitro and in vivo data for its PASylated hGH

FREISING, GERMANY, September 23, 2011 – XL-protein GmbH, a German biotech company specialised in the development of biopharmaceuticals with extended plasma half-life, announced today preclinical in vitro and in vivo data for its xl020 PAS-hGH program. The xl020 data demonstrate XL-protein’s ability to furnish an approved biological drug with improved in vivo stability and plasma half-life using its proprietary PASylation® technology, leading to enhanced efficacy in an established animal model.

The results of XL-protein’s xl020 studies will be presented at the R&D of Novel Protein Therapeutics and Post-Translational Modifications Conferences on Tuesday and Wednesday 27/28 September 2011 in Berlin, Germany, in a presentation entitled “PASylation: a biological alternative to PEG for extending the plasma half-life of biopharmaceuticals”.

“This program is extremely exciting because it provides an in vivo proof of concept that a PASylated biological does not only show prolonged pharmacokinetics but also increased potency, even if applied at longer time intervals compared with the unmodified protein,” stated Uli Binder, co-founder and CTO of XL-protein GmbH.

XL-protein researchers documented that xl020 has much longer plasma-half life than the unmodified recombinant growth hormone while retaining high receptor binding activity, which translated into increased in vivo efficacy. In a preclinical mouse model of growth hormone deficieny, xl020 administration showed continuous strong weight gain over at least 10 days.

Arne Skerra, co-founder and CEO of XL-protein GmbH, said: “Our PASylated hGH program highlights XL-protein’s strategy of developing superior therapeutics based on approved biopharmaceuticals. xl020 is a promising development candidate that addresses a multi-billion dollar market world-wide.”

hGH is a natural peptide hormone that regulates body growth and metabolism. Growth hormone deficiency (GHD) can occur in both pediatric and adult patients (GHDA). Furthermore, hGH is currently approved for several other indications, for example AIDS wasting and cachexia or short stature caused by Turner’s Syndrome or Prader-Willi syndrome. xl020 should allow less frequent and lower dosing, promising better tolerability and patient compliance, also benefiting from the inherent biodegradability of PAS polypeptides.

XL-protein’s proprietary PASylation® technology can be applied both to existing biologics, yielding ‘biobetters’, or to innovative therapeutic proteins or peptides, leading to tunable prolonged plasma half-life by a factor 10-100 as demonstrated in numerous animal studies. Thus, PASylation® offers a superior solution to a general problem in biological drug development.

European patent granted for XL-protein’s PASylation® technology

FREISING, GERMANY, MARCH 17th, 2011 – XL-protein GmbH, a German biotech company that specialises in the development of biopharmaceuticals with extended plasma half-life, today announced that the European Patent Office granted a core patent (EP2173890) for XL-protein’s PASylation® technology.

Beside the granted PASylation® EP patent, several other corresponding national patent applications are currently processed worldwide. The European patent covers the use of random coil polypeptide sequences comprising the natural amino acids Proline, Serine, and Alanine (PAS) to enhance the stability of biologically active proteins if attached as part of a fusion protein.

The N- or C-terminal PAS tag can be directly produced together with the therapeutic protein in microbial hosts, e.g. E. coli, or in cell culture, thus avoiding costly in vitro coupling steps that are required for other presently available approaches such as PEGylation. Furthermore, the biodegradability of the PAS polypeptide should prevent organ accumulation during chronic treatment.

PASylation® can be applied both to existing biologics, yielding ‘biobetters’, or to innovative therapeutic proteins or peptides, leading to a prolonged plasma half-life by a factor 10-100 as demonstrated in numerous animal studies. Thus, PASylation® offers a superior solution to a general problem in biological drug development, eventually allowing less frequent and lower dosing together with better tolerability for patients.

Dr. Arne Skerra, founder and CEO of XL-protein GmbH, said: “This patent is an important milestone for our company and its collaboration partners. It reinforces our competitive market position and will strengthen corporate development”.
This revolutionary principle for increasing drug stability was originally developed at the Technische Universität München (TUM). In 2009, XL-protein entered into a license agreement with TUM via Bayerische Patentallianz GmbH, the central patent and marketing agency for 28 Bavarian universities and universities of applied sciences, and acquired the exclusive worldwide rights for the PASylation® technology, which also includes the right to grant sublicenses.
Mr. Peer Biskup, CEO of the Bayerische Patentallianz GmbH, commented: “The Bayerische Patentallianz is very pleased with the timely decision of the EPO to grant the first PASylation® patent and with the positive development of XL-protein’s business, which involves both in house drug development activities and partnering with pharma and biotech companies.”

Phylogica collaborates with XL-protein on inflammation Program

XL-protein Collaborates with Phylogica on Inflammation Program

PERTH, AUSTRALIA, January 13, 2011 – Phylogica Ltd (ASX: PYC), a leading Australian-based drug discovery company, announced today an alliance with XL-protein GmbH, a German-based biotech company that specialises in extending the circulation half-life of biopharmaceuticals. Through this alliance, Phylogica gains access to a unique modification technology that strengthens the Company’s Phylomer drug discovery platform.

XL-protein’s PASylation technology was originally developed by a leading academic group at the prestigious Technical University of Munich (TUM). The technology utilises a genetic technique to fuse a polymer of natural amino acids onto the relevant peptide or protein-based therapeutic, which slows the body’s clearance of the drug and prolongs the therapeutic effect.

Phylogica and XL-protein will initially collaborate on a pilot study to validate the technology in combination with one of Phylogica’s Phylomer peptides targeting CD40 ligand from its in-house anti-inflammatory programme. Under the terms of the agreement, Phylogica has an option to obtain an exclusive license to the PASylation technology for peptide-based products in relation to CD40 ligand and has rights of negotiation for a license to the technology for Phylomers that bind other targets.

It is anticipated that the plasma half-life of PASylated Phylomers will be significantly extended, making them more suitable for less frequent administration and at lower dose. Long acting Phylomers would be ideal for treating chronic indications, such as rheumatoid arthritis, coronary artery disease and inflammatory bowel disease, in which CD40 ligand has been implicated as a potential target.

Phylogica’s Chief Executive Officer, Dr Paul Watt, commented: “Phylogica is excited to enter into this collaboration with XL-protein. This deal provides us with access to the next generation of polypeptide fusions for optimising the half-life of our Phylomers. The technology combination of XL-protein and Phylogica could also significantly facilitate the ease of manufacturing and lower the cost of goods.”

Dr Arne Skerra, founder and CEO of XL-protein GmbH, said: “XL-protein is delighted to apply its PASylation technology to Phylomers, which will serve to validate the benefit of this revolutionary half-life extension technology to a highly promising class of therapeutic peptide drugs.”

The Bavarian Patent Alliance GmbH and the biotech company XL-protein GmbH have signed an exclusive licensing agreement on a promising invention of the Technical University of Munich

München, Germany, September 17, 2009 The Bavarian Patent Alliance GmbH and the biotech company XL-protein GmbH have signed an exclusive licensing agreement on a promising invention of the Technical University of Munich.

With the so-called “PASylation technology” the plasma half-life of biological agents is extended manifold. Therapeutic proteins are directly fused on the genetic level with sequences of the amino acids proline, alanine and serine. The “PAS” polymers are stable in plasma and pharmacologically inert. They adopt a bulky random coil structure, which significantly increases the size of the protein. Thus, the typically rapid clearance of the biological via kidney filtration is retarded by 1-2 orders of magnitude. In this way the duration of drug action in the human body can be extended to several days.

Through the acquisition of the worldwide patent rights XL-protein takes, as the exclusive licensee, advantage of the PASylation technology originally developed at the Institute of Biological Chemistry at the Technical University of Munich (TUM).

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