Oral IL-10 Fusion Protein Formulation for Dimer Stability

Solution Overview

Problem

Oral administration of protein pharmaceuticals faces challenges such as denaturation in the stomach, hydrolysis by gastrointestinal enzymes, and difficulty crossing the intestinal epithelium due to large size, and common purification methods hinder proper dimer formation of therapeutic proteins.

Innovation Solution

Development of delivery constructs with specific amino acid sequences, such as SEQ ID NO: 5 or 13, formulated for oral administration, and methods involving refolding and chromatography to enrich for IL-10 delivery constructs in dimer form, using a composition with multiple copolymers to protect against acidic environments and enhance intestinal absorption.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If protein pharmaceuticals are administered orally, then convenience and patient compliance are improved, but the protein structure is denatured by acidic environment and proteolytic enzymes

Engineering Contradiction:

Improveconvenience of administrationVSAvoidprotein structure stability

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The patent uses a carrier protein (human serum albumin) as an intermediary to protect the therapeutic protein from denaturation in the acidic GI environment. The carrier forms a protective complex that shields the protein from harsh conditions during oral transit, enabling stable delivery while maintaining therapeutic efficacy.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent employs pH-responsive parameters by designing the delivery system to remain stable at acidic pH in the stomach but undergo controlled release at neutral pH in the intestine. This parameter change allows the protein to survive the acidic environment while being released at the target site.

Inventive Principle:

Principle #35Parameter changes

2Ease of manufacture

If common purification protocols are used, then protein production is simplified, but proper dimer formation is prevented resulting in high proportion of monomers or aggregates

Engineering Contradiction:

Improvepurification process simplicityVSAvoiddimer formation control

Core Design Contradiction:

Ease of manufactureVSManufacturing precision

Solution Approach 1:

The patent applies preliminary action by incorporating dimerization-promoting conditions into the purification protocol from the outset. The method includes specific buffer compositions and controlled incubation conditions that facilitate proper dimer formation during the purification process itself, rather than attempting to correct dimerization issues after purification.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent changes physical and chemical parameters during purification, including pH, ionic strength, and temperature, to optimize dimer formation. Specific buffers and additives are used to create conditions that favor dimer assembly while maintaining protein stability throughout the purification process.

Inventive Principle:

Principle #35Parameter changes

3Productivity

If therapeutic proteins are produced in monomer form, then production is easier, but therapeutic efficacy is compromised as dimers are more active

Engineering Contradiction:

Improveproduction efficiencyVSAvoidtherapeutic activity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies preliminary action by incorporating dimerization-promoting conditions into the purification protocol from the outset. The method includes specific buffer compositions and controlled incubation conditions that facilitate proper dimer formation during the purification process itself, rather than attempting to correct dimerization issues after purification.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent changes physical and chemical parameters during purification, including pH, ionic strength, and temperature, to optimize dimer formation. Specific buffers and additives are used to create conditions that favor dimer assembly while maintaining protein stability throughout the purification process.

Inventive Principle:

Principle #35Parameter changes

4Reliability

If large-sized protein pharmaceuticals are administered orally, then therapeutic effect is achieved, but crossing the intestinal epithelium becomes difficult

Engineering Contradiction:

Improvetherapeutic effectVSAvoidintestinal absorption

Core Design Contradiction:

ReliabilityVSEase of operation

Solution Approach 1:

The patent uses the carrier protein as an intermediary vehicle that facilitates intestinal epithelium crossing. The carrier-protein complex is recognized by intestinal transport mechanisms, enabling the large therapeutic protein to cross the epithelial barrier through carrier-mediated transport rather than passive diffusion.

Inventive Principle:

Principle #24Intermediary (Mediator)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The solution ensures high dimer formation and effective oral delivery of IL-10, achieving therapeutic efficacy by promoting transcytosis across the gut epithelium and maintaining activity in inflammatory diseases like ulcerative colitis and Crohn's disease.

Implementation Method 1

a first coat comprising a first copolymer and a second copolymer... configured to release substantially none of the IL-10 delivery construct after 1 h exposure to a solution having a pH of 1.0... configured to release at least 40% of the IL-10 delivery construct after 2 hours of exposure to a solution having a pH of 7.0

Methodology Applied

Scientific EffectAcidic environment protection:

Implementation Method 2

achieving therapeutic efficacy by promoting transcytosis across the gut epithelium

Methodology Applied

Scientific EffectTranscytosis:

Implementation Method 3

methods involving refolding and chromatography to enrich for IL-10 delivery constructs in dimer form

Methodology Applied

Scientific EffectChromatography: Chromatography

Implementation Method 4

methods involving refolding and chromatography to enrich for IL-10 delivery constructs in dimer form

Methodology Applied

Scientific EffectProtein refolding:

Data Source

PatentUS20260001927A1Compositions, formulations and interleukin production and purification

Publication Date: 2026.01.01 THORNHILL THERAPEUTICS INC

AI summary

Described herein are cholix-IL-10 fusion proteins, and methods of use thereof, which can be characterized by a distinct response in an individual when administered. This distinct response can comprise changes in levels of one or more markers in the individual and/or co-localization of IL-10 in the Lamina propria of the individual. Further described herein, in some embodiments, are oral formulations of the cholix-IL-10 fusion proteins. Described herein are methods for the purification of an IL-10 delivery construct, including methods for refolding and enrichment, which can result in maintenance of a high percentage of the IL-10 delivery constructs in the biologically active dimer form. Described herein are oral formulations configured for site-specific release of a therapeutic protein in the small intestines or colon. In some cases, the therapeutic protein is in the form of a dimer, such as an IL-10 delivery construct capable of crossing the gut epithelium.