Stable Cyanine Dyes for Near-Infrared Bioimaging

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Solution Overview

Problem

Existing cyanine dyes lack stability in the presence of common bioassay reagents and suffer from thermal and photostability issues, making them unsuitable for applications like DNA sequencing and in vivo imaging, and are often difficult to synthesize in high purity and yield due to asymmetry.

Innovation Solution

Development of stable and symmetric cyanine dyes with specific polymethine bridge structures and bioconjugates that can be used for labeling biomolecules and in vivo imaging, including compounds of Formula I and II, which are designed to be more robust and versatile.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional cyanine dyes are used for labeling biomolecules, then fluorescence imaging in the near-infrared region can be achieved, but the dyes lack stability in the presence of common bioassay reagents and have poor thermal and photostability

Engineering Contradiction:
Improvestability of cyanine dyesVSAvoidcompatibility with bioassay reagents
Core Design Contradiction:
ReliabilityVSAdaptability or versatility

Solution Approach 1:

The patent modifies the chemical structure of cyanine dyes by introducing specific substituents (such as sulfur-containing groups and cyclic structures) to enhance stability. The polymethine bridge is designed with specific configurations (e.g., 5-membered rings with sulfur atoms) that increase resistance to reagents like ammonium hydroxide, DTT, and APS while maintaining near-infrared fluorescence properties.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The invention creates composite dye structures by combining multiple functional groups within the cyanine molecule - including heterocyclic rings, polymethine bridges, and stabilizing substituents - to achieve both stability and versatility. These composite structures integrate multiple properties: photostability, chemical stability, and biomolecule compatibility within a single molecular framework.

Inventive Principle:
Principle #40Composite materials

2Adaptability or versatility

If asymmetric cyanine dyes are synthesized, then structural diversity can be achieved, but synthesis difficulty increases and purity/yield decreases

Engineering Contradiction:
Improvestructural diversity of dyesVSAvoidsynthesis difficulty and purity
Core Design Contradiction:
Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent employs asymmetric substitution patterns on the cyanine core structure, where different substituents are placed at different positions of the polymethine bridge or heterocyclic rings. This controlled asymmetry generates structural diversity for various applications while maintaining synthetic feasibility through systematic design rules.

Inventive Principle:
Principle #4Asymmetry

Solution Approach 2:

The synthesis approach divides the complex cyanine dye synthesis into modular steps: first constructing the core polymethine bridge structure, then adding substituents in separate stages. This segmentation allows for better control over purity and yield while achieving diverse final structures.

Inventive Principle:
Principle #1Segmentation

3Object-affected harmful factors

If cyanine dyes are used for in vivo imaging, then near-infrared fluorescence can be achieved with reduced autofluorescence interference, but photostability and thermal stability are insufficient

Engineering Contradiction:
Improveautofluorescence interferenceVSAvoidphotostability and thermal stability
Core Design Contradiction:
Object-affected harmful factorsVSDuration of action of stationary object

Solution Approach 1:

The patent optimizes the conjugation length and substitution patterns of the polymethine bridge to shift emission wavelengths into the near-infrared region (600-1000 nm), where biological autofluorescence is minimal. Simultaneously, stabilizing groups are incorporated to enhance photostability, allowing prolonged imaging sessions without rapid signal degradation.

Inventive Principle:
Principle #35Parameter changes

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 new cyanine dyes provide enhanced stability and photostability, enabling effective labeling and imaging applications, including improved performance in Western blotting and in vivo imaging, with balanced charge and suitable absorption and emission spectra for biomedical use.

Implementation Method 1

many of these dyes fluoresce in the near-infrared (NIR) region of the spectrum (600-1000 nm)

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

cyanine dyes strongly absorb and fluoresce light

Methodology Applied
Scientific EffectAbsorption of light: Absorption (EM radiation)

Data Source

PatentEP2419478B1Fluorescent imaging with substituted cyanine dyes
Publication Date: 2018.02.28 LI COR INC
  • EP2419478B1 patent drawingFigure 1A~1D
  • EP2419478B1 patent drawingFigure 2A~2D
  • EP2419478B1 patent drawingFigure 3A~3B

AI summary

Compounds and methods are disclosed that are useful for noninvasive imaging in the near-infrared spectral range. The cyanine compounds of Formula (I) are presented: (I). wherein Q is a portion of a polymethine bridge selected from the group consisting of: (a) and (b). Also included are bioconjugates of the compounds of Formula (I), methods of labeling biomolecules with the compounds, and methods of imaging.