Endostatin Mutant ATPase Activity Enhancement

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Solution Overview

Problem

Native endostatin's ATPase activity is negatively related to its anti-tumor activity, limiting its effectiveness in inhibiting angiogenesis and tumor growth, as evidenced by decreased activity in mutants with lower ATPase activity.

Innovation Solution

Identification of an auxiliary ATP-binding motif in endostatin, Val-Leu-Cys-Ile-Glu, and strategic mutations in the Walker A and B motifs to enhance ATPase activity, resulting in ES mutants with increased anti-angiogenesis and anti-tumor activity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If native endostatin is used to inhibit angiogenesis and tumor growth, then anti-tumor activity is achieved, but ATPase activity limits its effectiveness

Engineering Contradiction:
Improveanti-tumor activityVSAvoidATPase activity
Core Design Contradiction:
ReliabilityVSUse of energy by moving object

Solution Approach 1:

The patent applies parameter changes by mutating specific amino acid residues in the ATP-binding motifs (Walker A and Walker B motifs) of endostatin to alter its ATPase activity. By changing the chemical parameters of the protein structure through site-directed mutagenesis, the invention achieves enhanced anti-tumor activity while controlling the energy-related ATPase parameter.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality by making specific localized changes to the ATP-binding motifs rather than modifying the entire protein structure. The mutations are concentrated in the Walker A (GXXGXXK) and Walker B (h E) motifs, which are specific local regions responsible for ATP binding and hydrolysis, thereby improving overall anti-tumor activity through targeted local modifications.

Inventive Principle:
Principle #3Local quality

2Reliability

If ATPase activity is increased to enhance anti-angiogenesis activity, then inhibition of endothelial cell migration is improved, but energy consumption increases

Engineering Contradiction:
Improveinhibition of endothelial cell migrationVSAvoidATPase activity
Core Design Contradiction:
ReliabilityVSUse of energy by moving object

Solution Approach 1:

The patent optimizes the relationship between ATPase activity and anti-angiogenesis function by changing the amino acid parameters in the ATP-binding motifs. The mutations are designed to achieve the optimal balance where enhanced inhibition of endothelial cell migration is accomplished with controlled energy consumption, rather than simply maximizing ATPase activity.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates multiple mutant variants of endostatin with different amino acid substitutions in the ATP-binding motifs. These copies (mutants) allow selection of the optimal variant that achieves the desired anti-angiogenesis activity with appropriate energy consumption characteristics, rather than relying on a single protein structure.

Inventive Principle:
Principle #26Copying

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

ES mutants with elevated ATPase activity demonstrate significantly improved inhibition of endothelial cell migration and tumor growth, offering enhanced therapeutic potential for angiogenesis-related diseases like tumors, obesity, and insulin resistance.

Implementation Method 1

ATPase, also named as adenosine triphosphatase, is a class of enzymes which can catalyze the hydrolysis of ATP to release energy

Methodology Applied
Scientific EffectATP hydrolysis: Hydrolysis

Implementation Method 2

ATPase, also named as adenosine triphosphatase, is a class of enzymes which can catalyze the hydrolysis of ATP

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP3269731B1Molecular design of recombinant protein drug
Publication Date: 2023.10.04 BEIJING PROTGEN
  • EP3269731B1 patent drawingFigure 1
  • EP3269731B1 patent drawingFigure 2(A)~2(C)
  • EP3269731B1 patent drawingFigure 3~5

AI summary

Provided is a mutant of an endostatin. The mutant has improved ATPase activity and improved activity of inhibiting angiogenesis and inhibiting tumors. Further provided is use of the mutant in treatment of angiogenesis related diseases such as tumors.