Engineered FGF1 and FGF2 Polypeptides for Protease Resistance

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Solution Overview

Problem

FGF-1 and FGF-2 proteins have low intrinsic stability and are susceptible to proteolytic degradation, particularly by thrombin, which limits their effectiveness in wound healing and other therapeutic applications.

Innovation Solution

Engineered FGF1 and FGF2 polypeptides with specific amino acid substitutions, such as Q41P, S48L, H94S, and K113N for FGF1, and Q65L, N111S, K128N, and K138E for FGF2, which enhance stability against proteolytic degradation and maintain heparin-independent mitogenic activity, resulting in hyper-stable variants like 'super hFGF1' and 'super hFGF2'.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If FGF-1 and FGF-2 proteins are used as therapeutics, then they exhibit mitogenic activity and promote wound healing, but they are highly susceptible to proteolytic degradation by thrombin, resulting in low intrinsic stability

Engineering Contradiction:

Improvestability against proteolytic degradationVSAvoidcirculation half-life

Core Design Contradiction:

ReliabilityVSDuration of action of stationary object

Solution Approach 1:

The patent applies parameter changes by introducing specific amino acid substitutions at defined positions in the FGF-1 and FGF-2 protein sequences. These substitutions modify the protein's structural parameters to resist thrombin cleavage while preserving biological activity, thereby extending circulation half-life and improving stability without compromising therapeutic function

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent converts the harmful effect of thrombin (which normally degrades FGF proteins) into a beneficial selection criterion. By designing proteins with specific resistance mutations, the invention transforms the threat of proteolytic degradation into an opportunity to create therapeutics that are specifically resistant to thrombin, turning a harmful interaction into a protective feature

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

2Productivity

If FGF-1 and FGF-2 proteins are administered for wound healing, then they stimulate cell proliferation and migration, but they are rapidly degraded by thrombin present in fibrin clots at the wound site

Engineering Contradiction:

Improvemitogenic activityVSAvoidstability in wound environment

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent modifies the protein parameters through targeted amino acid substitutions that specifically address thrombin degradation. These changes allow the FGF proteins to maintain their mitogenic activity in the wound environment while gaining resistance to thrombin-mediated degradation, thus improving stability without losing productivity

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent segments the protein structure at specific positions (Q41, S48, H94, K113 for FGF-1; Q65, N111, K128, K138 for FGF-2) to introduce resistance mutations. This segmentation allows selective modification of specific regions responsible for thrombin interaction while preserving the overall protein function and activity

Inventive Principle:

Principle #1Segmentation

Data Source

PatentUS11267855B2Engineered FGF1 and FGF2 compositions and methods of use thereof

Publication Date: 2022.03.08 THE BOARD OF TRUSTEES OF THE UNIV OF ARKANSAS

AI summary

Engineered FGF1 and FGF2 polypeptides, polynucleotides encoding these polypeptides and DNA constructs, vectors and compositions including these engineered polypeptides are provided herein. The engineered FGF1 and FGF2 polypeptides are more stable than their wild-type counterparts and may be more effective at treating a variety of conditions that FGF1 and FGF2 are useful for treating such as wound healing.