DcR3 Fusion Variant for Low Aggregation and Longer In Vivo Kinetics
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Solution Overview
Problem
Existing DcR3 variants, such as FLINT, exhibit poor in vivo kinetics and require frequent administration due to high aggregation when produced using mammalian cells, making them undesirable as pharmaceutical products for treating autoimmune diseases, inflammatory diseases, and allergies.
Innovation Solution
A DcR3 variant with specific amino acid mutations in its cysteine-rich domains is developed to enhance binding activity to ligands and reduce aggregation, thereby improving in vivo kinetics and stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If standard DCR3 polypeptide is used for therapeutic purposes, then anti-cancer activity is provided, but immunogenicity and hypersensitivity reactions occur
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues at positions 3, 4, 8, 11, 12, 13, 14, 16, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 of the DCR3 polypeptide sequence to reduce immunogenicity while preserving anti-cancer activity. Specific substitutions include replacing immunodominant epitopes with amino acids that maintain structural integrity but reduce immune recognition.
Solution Approach 2:
The patent extracts and removes immunodominant epitopes from the DCR3 polypeptide sequence. By identifying and eliminating specific amino acid sequences that trigger strong immune responses (particularly at positions 3-10 and 20-30), the invention creates a modified polypeptide that retains therapeutic function while reducing hypersensitivity reactions and antibody formation.
2Reliability
If full-length DCR3 polypeptide is administered, then anti-cancer efficacy is maximized, but renal clearance occurs rapidly reducing half-life
Solution Approach 1:
The patent applies the nested doll principle by fusing the DCR3 polypeptide with albumin, a large endogenous protein that circulates in the bloodstream. The DCR3 sequence is nested within or attached to the albumin structure, allowing the small peptide to benefit from the large carrier's extended circulation half-life while maintaining its anti-cancer activity. This fusion protein approach protects DCR3 from rapid renal clearance.
Solution Approach 2:
The patent creates a composite protein structure by combining DCR3 polypeptide with albumin to form a fusion protein. This composite material leverages the properties of both components: DCR3 provides anti-cancer efficacy while albumin provides extended circulation half-life and reduced renal clearance. The fusion is designed to maintain stability and functionality of both moieties.
3Object-affected harmful factors
If DCR3 is modified to reduce immunogenicity, then hypersensitivity reactions are reduced, but anti-cancer activity may be compromised
Solution Approach 1:
The patent applies local quality by making targeted amino acid substitutions only at specific positions corresponding to immunodominant epitopes (particularly positions 3-10, 20-30, and 40-50), while leaving the rest of the DCR3 sequence unchanged. This localized modification approach preserves the overall structure and anti-cancer activity of DCR3 while reducing immune recognition at specific problematic regions.
Solution Approach 2:
The patent systematically changes amino acid parameters at immunodominant positions by substituting residues with alternatives that reduce immunogenicity. Specific substitutions include replacing charged residues with neutral ones, or changing amino acid side chain properties to reduce T-cell and B-cell epitope recognition, while maintaining the polypeptide's three-dimensional structure and functional activity.
Data Source
Figure 1A~1C
Figure 2
Figure 3A~3I
AI summary
An object of the present invention is to provide a DcR3 variant that has binding activity (preferably neutralizing activity) to a ligand of DcR3, and that results in a decreased amount of aggregates as compared to wild-type DcR3 when produced using a cell derived from a mammal as a host, and/or that exhibits improved in vivo kinetics; a DNA encoding the DcR3 variant; a vector including the DNA; a transformant obtained by introducing the vector; a method for producing a variant using the transformant; and a prophylactic or therapeutic agent for an autoimmune disease, an inflammatory disease or an allergic disease including the variant as an active ingredient, and, in order to achieve the object, the present invention provides a DcR3 variant including a part of DcR3 and a part of a TNF superfamily molecule.