Tetrapeptide Synthesis via Segmented Amino Acid Mixtures

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Solution Overview

Problem

Existing methods for producing peptide combinations lack reproducibility in quality and quantity, which is essential for therapeutic applications.

Innovation Solution

A method involving two amino acid mixtures with identical molar ratios, where one mixture contains protected acid groups and the other contains protected amino groups, allowing for the synthesis of tetrapeptide combinations that are reproducibly qualitative and quantitative, using specific protecting groups and coupling reagents like Fmoc, Boc, and EDC, and subsequent removal of protective groups to form stable tetrapeptide libraries.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If statistical or partial hydrolysate methods are used to produce peptide combinations, then a diverse peptide mixture can be obtained, but the quality and quantity are not reproducible across different batches

Engineering Contradiction:
Improvereproducibility of peptide combination quality and quantityVSAvoidbatch-to-batch consistency
Core Design Contradiction:
Manufacturing precisionVSReliability

Solution Approach 1:

The synthesis process is divided into two separate amino acid mixtures (Mixture A with protected acid groups and Mixture B with protected amino groups) that are combined in a controlled manner. This segmentation allows independent optimization and characterization of each mixture before combination, ensuring reproducible results across batches.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention specifies precise molar ratios for amino acids in both mixtures (e.g., Gly: 30-70 mol%, Ala: 5-30 mol%, Val: 5-30 mol%, Leu: 5-30 mol%, Pro: 5-30 mol%) and controlled mixing ratios (e.g., 1:1 to 1:5 weight ratio of Mixture A to Mixture B). These parameter specifications ensure reproducible peptide combination quality and quantity across different production batches.

Inventive Principle:
Principle #35Parameter changes

2Ease of manufacture

If natural amino acids are used directly without protection, then the synthesis is simpler, but the reaction specificity and product purity are reduced

Engineering Contradiction:
Improvesimplicity of synthesis processVSAvoidproduct purity and reaction specificity
Core Design Contradiction:
Ease of manufactureVSManufacturing precision

Solution Approach 1:

Amino acids are pre-protected with protecting groups (Fmoc, Boc for amino groups; OtBu, OBzl for acid groups) before synthesis. This preliminary protection prevents unwanted side reactions and ensures that coupling reactions occur only at the desired positions, thereby improving product purity while maintaining manageable synthesis complexity through systematic deprotection steps.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

Protecting groups act as intermediaries that temporarily modify amino acid functionality to enable selective reactions. The protecting groups are introduced, reactions are performed with high specificity, and then the protecting groups are removed in a controlled manner to yield the desired pure peptide products.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Quantity of substance

If complete hydrolysis of thymus proteins is performed, then all peptide components are released, but the process is time-consuming and may lead to loss of labile peptides

Engineering Contradiction:
Improvecompleteness of peptide extractionVSAvoidhydrolysis time
Core Design Contradiction:
Quantity of substanceVSLoss of time

Solution Approach 1:

Instead of performing complete hydrolysis of thymus proteins, the invention pre-identifies and characterizes individual peptides from partial hydrolysates, then synthesizes these specific peptides from amino acid mixtures. This preliminary identification and targeted synthesis approach eliminates the need for time-consuming complete hydrolysis while ensuring all relevant peptide components are obtained with high purity and without loss of labile peptides.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The invention creates synthetic copies of naturally occurring thymus peptides by analyzing the amino acid composition and sequence of peptides from partial hydrolysates, then reproducing these peptides through controlled synthesis from amino acid mixtures. This copying approach is more efficient than complete hydrolysis and ensures reproducible production of the same peptide components.

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

This approach ensures consistent quality and quantity of tetrapeptide combinations, as demonstrated by chromatographic resolution of independently produced batches, making them suitable for therapeutic preparations.

Implementation Method 1

combining two amino acid mixtures A and B, which contain an identical number of amino acids whose molar ratios are adjustable—that is, by reacting two mixtures of amino acids identical in their amino acid composition, except for their protecting groups

Methodology Applied
Scientific EffectChemical coupling: Chemical Bonding

Implementation Method 2

subsequent removal of protective groups to form stable tetrapeptide libraries

Methodology Applied
Scientific EffectDeprotection:

Data Source

PatentEP2235038B1Tetrapeptide Combination
Publication Date: 2019.06.05 GKL BIOTEC
  • EP2235038B1 patent drawingFigure 1
  • EP2235038B1 patent drawingFigure 2

AI summary

The invention relates to a peptide combination characterized by peptides each having the same sequence length (SEQL), that can be produced from a mixture (A) comprising a number x of amino acids having a protected acid group or a number z of peptides having an acid group protected by means of a protecting group and an activated amino group, wherein the amino acids are present in the mixture (A) in particular adjustable molar ratios, and a mixture (B) comprising a number y of amino acids having an amino group protected by means of a protecting group, wherein the amino acid molar ratios of the mixture (B) are equal to the amino acid molar ratios of the mixture (A), and wherein the number x=y.