XL-protein announces publication of key scientific data on its PASylation® technology

FREISING, GERMANY, August 30, 2013 – XL-protein GmbH announced today that key scientific data have been published in the journal Protein Engineering, Design & Selection (2013, Vol. 26, No. 8, pp. 489–501, 2013). The Open Access Publication “PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins” is also available for download at XL-protein’s web site (http://www.xl-protein.com/publications.html).

A major limitation of biopharmaceutical proteins is their fast clearance from circulation via kidney filtration which strongly hampers efficacy in human therapy. XL-protein has developed conformationally disordered polypeptide chains with expanded hydrodynamic volume comprising the small residues Pro, Ala and/or Ser (PAS). PAS sequences are hydrophilic, uncharged, genetically encodable amino acid polymers with biophysical properties very similar to poly-ethylene glycol (PEG), whose conjugation to drugs is a well known method for plasma half-life extension.

In contrast, beside chemical coupling PAS polypeptides offer fusion to a therapeutic protein on the genetic level, permitting production of fully active proteins in E. coli and other widely used host organisms (including cell culture secretion) without any in vitro modification steps. Importantly, PAS polypeptides are biodegradable, thus avoiding organ accumulation, while showing stability in serum and lacking toxicity or immunogenicity in animals. The publication describes that PASylation® furnishes typical biologics, such as interferon, growth hormone or antibody Fab fragments, with considerably prolonged circulation in vivo.

This work is complemented by another publication “High-yield production of PASylated human growth hormone (hGH) using secretory E. coli technology” that has appeared in the journal BioProcess International (2013, Vol. 11, No. 4, pp. 30–38) and is also available for download at XL-protein’s web site.

“PASylation® offers unique advantages, that is, surprisingly similar biophysical behavior compared with PEGylation, strong and tunable PK extending effects and conservation of high target-binding activity,” stated Uli Binder, CTO of XL-protein GmbH.

Arne Skerra, CEO of XL-protein GmbH, said: “PASylation® enables the preparation of biologically and/or pharmaceutically functional proteins with prolonged and enhanced in vivo activity, which constitutes a bottleneck in current biological drug development and opens exciting commercial opportunities.”

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 up to now.
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XL-protein and GENERIUM sign broad therapeutic license and collaboration agreement to develop PASylated therapeutics for the Russian Federation

Freising, Germany & Moscow, Russia, June 25, 2013

XL-protein and GENERIUM announce the closing of a license and a collaboration agreement. Under these agreements, XL-protein will license a PASylated blood clotting factor and apply its proprietary PASylation® technology for plasma-half life extension to a cytokine, respectively, for use in the Russian Federation and Commonwealth of Independent States (CIS).

GENERIUM acquires exclusive marketing rights for the Russian Federation and CIS and will assume responsibility for further development and marketing of said compounds within this territory. XL-protein retains development and marketing rights for Rest of World.

Under the terms of the license agreement, XL-protein receives a seven digit US$ upfront payment upon signing of the agreement. In addition, XL-protein will receive payments for the achievement of preclinical, clinical, regulatory, and commercial milestones as well as significant royalties on sales.

“This strategic cooperation with a renowned pharmaceutical company of GENERIUM’s caliber demonstrates the high potential of our PASylation® technology to develop biologics with superior activities”, noted Prof. Dr. Arne Skerra, CEO of XL-protein.

“Setting up multiple international collaborations is a key element of Generium’s strategy. The Company provides unique opportunities for international collaborative R&D projects which are often associated with technology and know-how transfer and are actively supported by the Russian Ministry of Industry and Trade”, noted PhD Sergey Ruchko, CEO of IBC Generium.

XL-protein’s proprietary PASylation® technology is a biological alternative to PEGylation, conferring an expanded hydrodynamic volume onto the biopharma-ceutical which leads to retarded kidney filtration based on a molecular size effect. A tunable prolonged plasma half-life by a factor 10-100 has been demonstrated for various compounds in preclinical animal studies. Thus, PASylation® offers a cost-effective and patient-friendly solution to a general problem in biopharmaceutical drug development.

First data from this collaboration will be presented at the 38th Congress of the Federation of European Biochemical Societies (FEBS) in St. Petersburg, Russia, July 6-11, 2013.

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XL-protein and Wacker Biotech produce a PASylated antibody fragment (Fab) with prolonged plasma half-life at more than four grams per liter culture in an E. coli expression system

High Yield with ESETEC® and PASylation®: Wacker Biotech and XL-protein Produce More than Four Grams per Liter of Fab Antibody Fragment with Prolonged Plasma Half-Life

Munich, Jena and Freising, April 18, 2013 – In the course of their
collaboration, Wacker Biotech and XL-protein have produced record yields of a PASylated Fab antibody using WACKER’s E. coli based ESETEC® secretion technology. WACKER developed an efficient bacterial cell line and was able to produce the Fab antibody in concentrations of more than four grams per liter of culture broth. This Fab antibody is being tested as a novel therapeutic agent for treating autoimmune diseases. These results open up new opportunities for developing monovalent antibody drugs that can be produced at lower cost, with fewer side-effects and with a tunable plasma half-life. As part of their recent collaboration agreement, WACKER and XL-protein are offering pharmaceutical customers access to the combination of the PASylation® platform and the ESETEC® technology. WACKER also has the necessary expertise for GMP-compliant production of biologics.

In a feasibility study, Wacker Biotech and XL-protein tested the use of WACKER’s patented ESETEC® technology to produce PASylated Fab antibody fragments for the development of new drugs to treat autoimmune

diseases. The study not only confirmed that the biopharmaceutical can be produced in high yield (4.3 g/l) by the patented ESETEC® technology. It also demonstrated that correctly folded, fully functional antibody fragments are efficiently secreted into the culture broth – this greatly simplifies downstream purification. Detailed biochemical characterization revealed monodispersity of the product and excellent antigen-binding activity.

For many years, antibody products have been a promising growth area for the pharmaceutical industry. Antibodies are highly specific in their ability to intervene with disease mechanisms, and they usually circulate in the body for several weeks. However, in certain indications, some of their features can be detrimental: stimulation of the immune system via the antibody effector region; bivalent binding or crosslinking of the antigen; or the antibodies’ long biological half-life. XL-protein therefore decided to develop a monovalent antigen-binding antibody fragment (Fab) that does not bind immunoreceptors and promises fewer sideeffects. Furthermore, PASylation® prolongs its plasma half-life and thus allows for optimal tuning of drug activity. Fab fragments are derived from human antibodies and can be manufactured as separate recombinant proteins. One of their advantages over full-length antibodies is an improved ability to penetrate the diseased tissue. Since Fabs are not glycosylated, they can be produced cost-effectively in microbial organisms such as E. coli. Nonetheless, Fab antibody fragments are much more difficult to produce than other therapeutic proteins, as they are composed of two different protein subunits and contain several disulfide bridges. So far, conventional production methods have yielded less than 2 grams per liter of culture.

“We are delighted – the results open up new avenues for the development of antibody products,” says Dr. Thomas Maier, managing director of Wacker Biotech. “ESETEC® produces Fab antibody fragments in record yields that until now had only been possible for whole antibodies using mammalian cell cultures. Yet, development and production times with ESETEC® are much faster than with mammalian cells. This saves our customers costs and shortens the time to market.” ESETEC® is a proprietary WACKER technology with a track record of cost-effective production of proteins and antibody fragments. It is based on an E. coli K12 strain which has the ability to secrete correctly folded recombinant proteins into the culture broth during fermentation. Secretion facilitates purification of the target protein, since there is no longer any need for complicated process steps such as homogenization and refolding. This makes the entire manufacturing process significantly more efficient and cost-effective. A number of biologics that have been manufactured with ESETEC® are already being evaluated in preclinical and clinical studies. Plus, a successful preliminary study with XL-protein has shown that the WACKER technology is highly efficient at producing PASylated human growth hormone.
According to Prof. Arne Skerra, managing director of XL-protein GmbH, “The fact that our PASylation® technology allows efficient and inexpensive production of antibody fragments with extended plasma half-life abolishes a major disadvantage of these highly effective and low-side-effect biologics, compared to conventional antibodies. Especially in conjunction with ESETEC®, it is now possible to rapidly manufacture improved antibody products with tailored properties in high yield.”

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Wacker Biotech and XL-protein Sign Long-Term Cooperation Agreement to Produce PASylated Biopharmaceuticals

Munich / Freising / Jena, October 30, 2012

Wacker Biotech GmbH and XL-protein GmbH are to collaborate more intensely on the production of PASylated therapeutic proteins. Today, the two companies announced the signature of an agreement to this effect. Through this collaboration, WACKER and its customers will gain access to XL-protein’s PASylation® platform. The PASylation® technology enables the development of biopharmaceuticals with extended plasma half-lives, which require less frequent injection and thus are more patient friendly. In a recent feasibility study, PASylated human growth hormone was successfully produced in high yields using WACKER’s E. coli-based ESETEC® technology.

In a feasibility study, Wacker Biotech and XL-protein tested WACKER’s patented ESETEC® technology for the production of large-sized PASylated therapeutic proteins. The study showed that ESETEC® produced a PASylated human growth hormone – correctly folded and fully functional – in high yields (3 to 4 g/l). Moreover, WACKER successfully used its secretion technology to develop an E. coli cell line for manufacturing the PASylated hormone. The production process is easy to implement on an industrial scale.

“Following the successful feasibility study, we are very much looking forward to intensifying our collaboration with XL-protein,” said Dr. Thomas Maier, managing director of Wacker Biotech GmbH. “More and more pharmaceutical companies are looking for efficient ways of prolonging the therapeutic effect of biologics. With this cooperation agreement with XL-protein,” he continued, “we can now offer our customers feasibility studies for the production of PASylated variants of their therapeutic lead candidates. Thanks to our innovative ESETEC® technology, we can supply research quantities of PASylated active ingredients for pre-clinical studies within a couple of weeks.”

“After our positive experience with WACKER’s innovative secretion technology in producing PASylated therapeutic proteins,” said Prof. Arne Skerra, XL-protein’s co-founder and CEO, “we want to continue our collaboration in this field on a long-term basis. By partnering with Wacker Biotech, we can offer access to our innovative technology to a much wider circle of customers,” he emphasized. “As an experienced GMP manufacturer, WACKER can ensure that PASylated proteins are produced quickly and professionally.”

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 directly into the culture broth during fermentation. This facilitates the subsequent purification of the target protein, since complicated process steps such as homogenization and refolding are unnecessary. This makes the entire manufacturing process significantly more efficient and cost-effective. A number of biologics manufactured with ESETEC® are already being evaluated in preclinical and clinical trials.

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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