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Method for monitoring reaction in synthetic DNA coding compound

A reaction monitoring and compound technology, applied in organic compound libraries, combinatorial chemistry, chemical libraries, etc., can solve the problems of different structures, different molecular weights, and difficulty in accurately monitoring the conversion degree of target compounds in mixed systems. Easy-to-use effects

Pending Publication Date: 2020-01-07
HITGEN INC
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, in the process of DNA-encoded compound library, a huge number of compounds are produced through the reaction steps of mixing multiple samples (the number of compounds in the mixed sample is as high as hundreds of thousands or even tens of billions), and each compound has a different structure and molecular weight. It is difficult to detect the molecular weight of each compound and the change of molecular weight of each compound by LC-MS
At the same time, different structures have different polarities, and the transformation of the reaction cannot be detected by commonly used monitoring methods such as HPLC.
Therefore, when synthesizing DNA-encoded compounds, it is difficult to accurately monitor the conversion degree of the target compound in the mixed system due to the influence of LC-MS / HPLC instrument conditions and sample conditions.

Method used

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  • Method for monitoring reaction in synthetic DNA coding compound
  • Method for monitoring reaction in synthetic DNA coding compound
  • Method for monitoring reaction in synthetic DNA coding compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, DNA-NH Concentration-absorption standard curve drawing of solution

[0065] Take DNA-NH2 solution with an initial concentration of 0.5mmol / L, and prepare standard solutions with concentrations of 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50mmol / L. Add the acetone solution (50mmol / L) of the fluorescent reagent 4-phenylspiro[furan-2(3H), 1-dihydroisobenzofuran]-3,3′-dione (50mmol / L) in the amount of 10 times the substance, 5°C Incubate for 10 minutes. Detect fluorescence absorption at an excitation wavelength of 388nm and an emission wavelength of 475nm, and draw an absorption-concentration standard curve, such as figure 1 shown.

Embodiment 2

[0066] Example 2, the drawing of the concentration-absorption standard curve of DNA-NH2 solution in the mixed system

[0067] Get the DNA-NH2 solution and DNA-NH solution that initial concentration is 0.5mmol / L, mix and prepare DNA-NH2 ratio (mol) be 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90% solution. Add 50 times (substance amount) fluorescent reagent acetone solution (50mmol / L), and incubate at 40°C for 5 minutes. Detect fluorescence absorption at an excitation wavelength of 388nm and an emission wavelength of 475nm, and draw an absorption-concentration standard curve, such as figure 2 shown.

Embodiment 3

[0068] Example 3. Monitoring of the reaction process in the complex system of the DNA-encoded compound library

[0069] 1. Construct a standard curve using a single DNA with a similar backbone

[0070] Get the 1b (structural formula such as Figure 5 Shown) solution, prepared as a standard solution with a concentration of 0.05, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50mmol / L. Add 20 times (substance amount) fluorescent reagent acetone solution (50mmol / L), and incubate at room temperature for 10 minutes. Fluorescent absorption was detected at an excitation wavelength of 388nm and an emission wavelength of 475nm, and an absorption-concentration standard curve was drawn. Such as image 3 shown

[0071] 2. Single DNA reaction monitoring process (such as Figure 4 )

[0072] 2.1 1a to 1b reactions (eg Figure 5 )

[0073] Dissolve 1a in sodium borate buffer (pH=9.4, concentration 250mmol / L) to prepare a 1mmol / L solution, and dissolve 1a with tris(2-carboxyethyl)p...

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Abstract

The invention provides a method for monitoring reaction in a synthetic DNA coding compound. The method for monitoring reaction in the synthetic DNA coding compound provided by the invention applies the fluorescence labeling technology to synthetic DNA coding compounds. The method can quickly, accurately, and efficiently monitor the degree of reaction conversion of a synthetic DNA coding compound mixing system.

Description

technical field [0001] In particular, the present invention relates to a reaction monitoring method in the synthesis of DNA-encoded compounds. Background technique [0002] In drug development, especially new drug development, high-throughput screening for biological targets is one of the main means to quickly obtain lead compounds. However, the traditional high-throughput screening based on a single molecule requires a long time, huge investment in equipment, limited number of library compounds (millions), and the construction of a compound library requires decades of accumulation, which limits the efficiency and efficiency of lead compound discovery. possibility. The DNA-encoded compound library synthesis technology that has emerged in recent years combines combinatorial chemistry and molecular biology techniques to add a DNA tag to each compound at the molecular level, and can synthesize hundreds of millions of compound libraries in a very short period of time. Moreover...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/64C40B40/08
Inventor 李进张帅何窦万金桥刘观赛
Owner HITGEN INC
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