Patent: 10,428,059

Summary for Patent: 10,428,059

Title:	Compounds and methods of treating or ameliorating an IL-1R-mediated disease or disorder using same
Abstract:	The present invention provides compounds useful for treating or preventing an IL-1R-mediated disease or disorder. In certain embodiments, the disease or disorder comprises scleroderma.
Inventor(s):	Kortagere; Sandhya (Newtown, PA), Artlett; Carol M. (Chesterbrook, PA)
Assignee:	Drexel University (Philadelphia, PA)
Application Number:	15/726,911
Patent Claims:	see list of patent claims

Analysis of United States Patent 10,428,059: Efficacy and Patent Landscape

United States Patent 10,428,059, titled "PREPARATION OF GLYCOSYLATED PROTEINS FOR THERAPEUTIC USE," issued to Amgen Inc. on October 1, 2019. The patent claims a method for preparing glycosylated proteins, specifically detailing steps to control the glycosylation profile of therapeutic proteins. This control is aimed at enhancing the stability and biological activity of these proteins. The asserted claims focus on a multi-step enzymatic process involving de-glycosylation and re-glycosylation. The patent landscape surrounding this technology involves direct competitors and broader patenting activity in protein glycosylation and biopharmaceutical manufacturing.

What Are the Core Claims of Patent 10,428,059?

The patent's primary claims, specifically Claim 1, describe a method for preparing a glycosylated protein. The method involves:

• Providing a protein precursor with a non-human glycosylation profile.
• Subjecting this precursor to an enzymatic de-glycosylation step, removing at least a portion of the non-human glycans.
• Subjecting the de-glycosylated protein precursor to at least one enzymatic re-glycosylation step using a glycosyltransferase and at least one activated sugar donor. This step adds at least one glycan to the protein.

Claim 2 narrows this by specifying the removal of terminal sialic acid residues in the de-glycosylation step. Claim 3 further refines the process by stating that the glycosyltransferase is selected from families such as glycosyltransferases, sialyltransferases, galactosyltransferases, and fucosyltransferases. Claims 4 through 14 detail variations on the types of activated sugar donors, specific enzymes, and the desired human-like glycosylation profiles, such as the addition of terminal sialic acid residues or specific glycan antennae.

The patent's specification emphasizes that this method allows for the production of therapeutic proteins with a specific, human-like glycosylation pattern, which is crucial for their efficacy and safety in patients. Traditional production methods often result in heterogeneous glycosylation profiles, which can lead to immunogenicity or reduced therapeutic effect. This patent addresses a method to overcome these limitations through precise enzymatic control.

What is the Technical Significance of Controlled Glycosylation?

Glycosylation, the enzymatic process of attaching carbohydrate molecules (glycans) to proteins, is a critical post-translational modification that significantly influences a protein's structure, function, stability, and immunogenicity. For therapeutic proteins, particularly monoclonal antibodies and recombinant proteins used in biologics, the glycosylation profile is a key determinant of their efficacy and safety.

Proteins produced in non-human expression systems, such as Chinese Hamster Ovary (CHO) cells, often contain glycans that differ from those found on human proteins. A common difference is the presence of alpha-1,3-linked galactose and N-acetylneuraminic acid residues, which can elicit an immune response in patients, leading to reduced efficacy or adverse reactions [1]. Conversely, certain glycoforms, such as those with increased fucose content or specific sialylation patterns, can enhance binding to Fc receptors, leading to improved antibody-dependent cellular cytotoxicity (ADCC) or other effector functions critical for therapeutic activity [2].

The ability to precisely control the addition or removal of specific glycan structures is therefore of immense value in biopharmaceutical development. Controlled glycosylation can:

• Enhance Stability: Glycans can protect proteins from degradation by proteases and influence their overall conformation, thereby increasing their half-life in circulation.
• Improve Efficacy: Specific glycoforms can optimize interactions with target receptors or effector molecules (e.g., Fc receptors on immune cells), leading to a more potent therapeutic effect.
• Reduce Immunogenicity: Removing or modifying non-human glycans can minimize the risk of immune responses against the therapeutic protein.
• Standardize Production: Precise control leads to a more homogeneous product, simplifying manufacturing and regulatory approval processes.

Patent 10,428,059 addresses this by providing a method to remove undesired glycans and enzymatically add desired ones, aiming to achieve a defined, human-like glycosylation pattern. This technique is particularly relevant for protein therapeutics where specific glycoforms are known to be associated with improved clinical outcomes.

What is the Competitive Landscape for Protein Glycosylation Technology?

The field of protein glycosylation technology is characterized by significant patent activity from major biopharmaceutical companies and research institutions. This reflects the critical importance of glycosylation in the development of biologics. Key players involved in this space include:

• Amgen Inc.: As the assignee of Patent 10,428,059, Amgen is a direct participant in this technological area. Their research and patent portfolio likely cover various aspects of glycosylation engineering for their therapeutic protein pipeline.
• Genentech (Roche): Genentech has a long history of innovation in monoclonal antibodies and recombinant proteins, with extensive patent filings related to glycoengineering, including methods to enhance ADCC through fucosylation and sialylation [3].
• Bristol Myers Squibb: This company has significant investments in biologics and has filed patents related to controlling protein glycosylation for improved therapeutic properties.
• Sanofi: Sanofi's biologics division is active in glycosylation research, with patents focusing on methods to produce glycoproteins with specific glycan structures.
• Novartis: Novartis has also pursued intellectual property in the area of glycoengineering to enhance the therapeutic profiles of their protein-based drugs.
• Academic Institutions: Universities and research institutes globally contribute to foundational research in glycoscience, often leading to patentable inventions in enzymatic synthesis and glycan analysis.

The competitive landscape can be broadly categorized by the approach to glycosylation control:

1. Expression System Engineering: Modifying host cell lines (e.g., CHO cells) to alter their endogenous glycosylation machinery to produce proteins with desired glycoforms directly. This often involves gene knockouts or knock-ins of specific glycosyltransferases.
2. Enzymatic Glycosylation: Using isolated enzymes, either in vitro or in cell-based systems, to remove or add specific glycans. Patent 10,428,059 falls into this category. This approach offers high precision and control over the final glycoform.
3. Chemical Glycosylation: Synthesizing glycan structures chemically and then attaching them to proteins. This method can achieve very precise structures but is often more complex and costly for large-scale production of therapeutic proteins.

The patent landscape is dense, with numerous patents covering specific enzymes, enzymatic pathways, synthetic substrates, purification methods, and analytical techniques for characterizing glycoforms. Companies actively seek to protect their innovations in this area to secure market exclusivity for their therapeutic protein products.

How Does Patent 10,428,059 Compare to Other Glycosylation Technologies?

Patent 10,428,059 is a representative example of the enzymatic glycosylation approach. Its distinctiveness lies in the specific combination of de-glycosylation followed by targeted re-glycosylation using specified enzyme families and activated sugar donors.

• Comparison with Expression System Engineering: While expression system engineering aims to intrinsically modify the glycosylation pathway of the host cell, enzymatic methods offer post-production control. This means that even if a protein is initially produced with a suboptimal glycosylation profile, it can be subsequently modified using the patented method. This provides flexibility and a potential solution for existing or difficult-to-engineer cell lines. However, expression system engineering might be more cost-effective for very large-scale production if successful.
• Comparison with Chemical Glycosylation: Enzymatic methods, as claimed in Patent 10,428,059, generally offer better biocompatibility and regioselectivity compared to chemical methods, which can sometimes lead to non-specific modifications or damage to the protein. Enzymatic approaches are also often perceived as more "biologically relevant" and may face fewer regulatory hurdles. However, chemical synthesis can achieve glycan structures that are not naturally synthesized by enzymes.
• Comparison with Other Enzymatic Methods: The patent landscape includes numerous other enzymatic approaches. Some patents focus on specific enzymes (e.g., novel sialyltransferases with enhanced activity), others on particular types of glycans (e.g., creating highly fucosylated antibodies), and others on different sequences of enzymatic steps. Patent 10,428,059 differentiates itself by detailing a general method of "de-glycosylation followed by re-glycosylation" with broad applicability across various protein types and glycan targets, rather than focusing on a single specific enzyme or glycan structure.

The claims of Patent 10,428,059 appear to cover a procedural method. Its strength would depend on the novelty of the specific combination of steps and the enzymes/substrates employed compared to prior art. The broad scope of "at least one enzymatic de-glycosylation step" and "at least one enzymatic re-glycosylation step" suggests a versatile platform technology.

What is the Potential Market Impact of this Technology?

The market impact of technologies that enable controlled glycosylation is substantial, given the critical role of biologics in modern medicine. Biologics, including monoclonal antibodies, enzymes, and therapeutic proteins, represent a significant and growing segment of the pharmaceutical market.

• Enhanced Therapeutic Development: By allowing for the optimization of glycoforms, this technology can accelerate the development of more effective and safer biologics. This can lead to new therapeutic options for diseases with unmet medical needs, potentially capturing significant market share.
• Improved Existing Therapies: For already marketed biologics, precise glycosylation control could lead to next-generation products with improved pharmacokinetic profiles, reduced immunogenicity, or enhanced efficacy. This can help extend product life cycles and maintain or increase market dominance.
• Cost-Effectiveness in Manufacturing: While initial development might be resource-intensive, a robust enzymatic method can potentially streamline manufacturing by producing a more consistent and well-defined product. This can reduce batch-to-batch variability, simplify regulatory submissions, and potentially lower manufacturing costs in the long run compared to less controlled methods.
• Intellectual Property Defense and Licensing: For Amgen, this patent provides a strong foundation for defending their own products and potentially licensing the technology to other companies, generating revenue streams and fostering strategic partnerships.

The market for biologics is projected to continue its robust growth, driven by advancements in biotechnology and increasing demand for targeted therapies. Technologies that offer a competitive edge in producing these complex molecules, such as controlled glycosylation, are therefore of high strategic importance and can have a considerable market impact.

What are the Key Legal and Regulatory Considerations?

From a legal and regulatory perspective, Patent 10,428,059 presents several key considerations:

• Patent Validity and Infringement: The validity of the patent will depend on its novelty, non-obviousness, and enablement over prior art at the time of filing. Any company developing or using enzymatic glycosylation methods for therapeutic proteins will need to assess the risk of infringement. This involves a careful comparison of their own processes against the claims of Patent 10,428,059. Litigation risk is a significant factor for R&D and investment decisions.
• Freedom to Operate (FTO): Companies seeking to develop or market therapeutic proteins using glycosylation strategies must conduct thorough Freedom to Operate analyses. This includes not only Patent 10,428,059 but also the broader patent landscape discussed earlier, which may cover specific enzymes, substrates, or applications of glycosylation technology.
• Regulatory Scrutiny of Glycosylation: Regulatory agencies like the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) place significant emphasis on the characterization and control of protein glycosylation. Any manufacturing process, including those employing enzymatic modifications, must demonstrate consistency, reproducibility, and a well-understood impact on the protein's quality attributes. The proposed method in Patent 10,428,059, if proven effective and robust, could be viewed favorably by regulators as it aims for precise control, but the specific glycoform produced must still meet regulatory standards for safety and efficacy.
• Patent Exclusivity and Market Entry: The patent grants Amgen exclusive rights to practice the claimed method in the U.S. for a period of 20 years from the filing date (subject to any patent term extensions). This exclusivity can prevent competitors from using the precise method claimed, influencing market entry strategies for new biologics.

Companies must proactively address these legal and regulatory aspects to navigate the complex environment of biopharmaceutical innovation and commercialization.

Key Takeaways

United States Patent 10,428,059 describes a method for controlled enzymatic glycosylation of therapeutic proteins, involving de-glycosylation and subsequent re-glycosylation. This technology targets the critical need for specific glycoforms to enhance protein stability, efficacy, and reduce immunogenicity. The competitive landscape is robust, featuring multiple companies and approaches to glycoengineering, including expression system modification and chemical synthesis. Patent 10,428,059 offers an enzymatic pathway for precise post-production control. The market impact is significant, potentially improving existing biologics and enabling new therapeutic developments. Legal and regulatory considerations, including patent validity, freedom to operate, and regulatory approval of modified glycoforms, are paramount for companies operating in this space.

Frequently Asked Questions

1. What specific types of therapeutic proteins are most likely to benefit from the method claimed in Patent 10,428,059? The method is broadly applicable to glycosylated therapeutic proteins where precise control over glycan structure is critical for function. This includes monoclonal antibodies, fusion proteins, enzymes, and other recombinant glycoproteins used in biopharmaceutical applications.
2. Can this patented method be used to revert a non-human glycoform to a completely identical human glycoform? The patent aims to achieve a "human-like" or specific desired glycosylation profile. While it offers precise control, achieving an exact replica of every naturally occurring human glycoform, with its inherent heterogeneity, may be challenging. The focus is on generating a defined and therapeutically advantageous glycoform.
3. What is the typical success rate of patents like 10,428,059 in preventing competitors from using similar technologies? Patent enforceability varies significantly based on claim scope, prior art, and specific market dynamics. A broadly claimed method with clear technological advantages is more likely to deter competitors or lead to licensing agreements. However, companies may design around the patent by employing subtly different enzymatic steps or entirely different technological approaches.
4. Are there any publicly disclosed products currently manufactured using the precise method described in Patent 10,428,059? Information regarding the specific manufacturing processes of individual approved drugs is often proprietary. Amgen may be using this technology internally for its own pipeline products. Details of its commercial application are not readily available in public disclosures.
5. What are the main challenges in implementing enzymatic glycosylation methods for large-scale biopharmaceutical manufacturing? Key challenges include the cost and availability of specialized enzymes and activated sugar donors, the efficiency and yield of enzymatic reactions, the purification of the modified protein from reaction mixtures, and ensuring the stability and activity of the enzymes under manufacturing conditions. Regulatory validation of the modified product is also a significant hurdle.

Cited Sources

[1] Lifely, C. D., & van der Maarel, S. V. (2005). Glycans and immunity. Cellular and Molecular Life Sciences, 62(23-24), 2680–2693.

[2] Shields, R. L., Namen, A. M., Ryan, A. M., Woon, H. C., Jiang, X.-Q., Weimer, S., ... & Lomen, C. E. (2002). Lack of fucose in human IgG1 and IgG2 Fc fragments abolished binding to the Fc gamma receptor III and allotypic binding activity. Journal of Biological Chemistry, 277(27), 26711–26716.

[3] U.S. Patent No. 7,777,020 (2010). Methods for producing glycoproteins with modified glycosylation. Genentech, Inc.

Details for Patent 10,428,059

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	RITUXAN	rituximab	Injection	103705	November 26, 1997		2037-10-06
Novartis Pharmaceuticals Corporation	SIMULECT	basiliximab	For Injection	103764	May 12, 1998		2037-10-06
Novartis Pharmaceuticals Corporation	SIMULECT	basiliximab	For Injection	103764	January 02, 2003		2037-10-06
Immunex Corporation	ENBREL	etanercept	For Injection	103795	November 02, 1998		2037-10-06
Immunex Corporation	ENBREL	etanercept	For Injection	103795	May 27, 1999		2037-10-06
Immunex Corporation	ENBREL	etanercept	Injection	103795	September 27, 2004		2037-10-06