Alkaline buffering solution for establishing DNA coding compound library and application thereof

A compound library and buffer technology, applied in the fields of sugar derivatives, sugar derivatives, organic chemistry, etc., can solve the problem of no detailed study on the reaction effect of single-tube operation, few references to buffer, and obvious amplification effect of batch operation And other issues

Active Publication Date: 2019-03-12
上海药明康德新药开发有限公司 +1
View PDF4 Cites 33 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current literature and patents related to the development of DNA-encoded compound libraries or on-DNA chemical reactions rarely mention the preparation of the buffer used, and there are almost no reports on the quality inspection of the buffer. Many on-DNA chemical reactions The development of the company only reported the single-tube operation method, and did not study in detail the steps and actual reaction effects of the single-tube operation method when it is actually applied to batch operation on a 96-well plate, and the 96-well plate operation is the DNA-encoded compound library. The soul of the operation
With regard to the above-mentioned buffer solution provided in the prior art, it has the following disadvantages: 1) the substrate applicability of the buffer solution is narrow, and the versatility is low, and a kind of buffer solution can only be applicable to the reaction of one or several reaction substrates; 2) When the single-tube operation method of the buffer solution is directly applied to batch operations on 96-well plates, inconsistent and large differences in reaction effects often occur, which has the disadvantage of obvious amplification effect of batch operations

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Alkaline buffering solution for establishing DNA coding compound library and application thereof
  • Alkaline buffering solution for establishing DNA coding compound library and application thereof
  • Alkaline buffering solution for establishing DNA coding compound library and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0142] Embodiment 1, 250mM, the preparation of the boric acid buffer solution of pH=9.4

[0143] 1. Preparation and quality inspection method of 250mM, pH=9.4 boric acid buffer solution

[0144] The specific preparation method is as follows:

[0145] Use an electronic balance with a precision of 0.01g to weigh Na according to Table 1 2 B 4 o 7 10H 2 O (23.8g, 0.062mol) was added to a clean 1L beaker, and 905g of deionized water was added to the above beaker, and after being stirred with a glass rod, it became a white suspension. Use 5M NaOH aqueous solution to adjust the pH value of the above solution to pH=9.4±0.1, and record the actual amount V1 (unit: mL), about 5.55 mL is needed. At this time, the solution should be a clear, transparent, colorless liquid, add deionized water M =73.7-V1 (unit: g) into the above-mentioned beaker, fully stirred and mixed, and filtered with a 0.22 μm filter into a sterile storage bottle.

[0146] 2. The quality inspection method of 250mM...

Embodiment 2

[0148] Example 2, alkaline boric acid buffer in DNA-CO 2 Application of H in Condensation Reaction of Small Molecule Organic Amines

[0149] According to the different reaction containers and operations, it is divided into single centrifuge tube and 96-well plate operation and introduced respectively.

[0150] 1. The specific operation steps of a single centrifuge tube are as follows:

[0151] 1) DNA-CO 2 H prepares active ester in situ with alkaline buffer and condensing agent

[0152]

[0153] DNA-CO 2 H (obtained from the hydrolysis of the initial head fragment mentioned in our previous patent CN108070009A and 4-methoxycarbonylbenzoic acid after condensation) was dissolved in 250mM, pH=9.4 boric acid buffer solution, prepared into a 1mM aqueous solution, and stored in 0.5 mL centrifuge tube (5μL, 5nmol), add 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.5μL, 500nmol , 200mM DMAc solution, 100 molar equivalents) and diisopropylethylami...

Embodiment 3

[0173] Embodiment 3, alkaline boric acid buffer solution in DNA-NH 2 Application in Reductive Amination Reaction with Small Molecular Organic Aldehydes

[0174] According to the different reaction containers and operations, it is divided into single centrifuge tube and 96-well plate operation and introduced respectively.

[0175] 1. The specific operation steps of a single centrifuge tube are as follows:

[0176] 1) DNA-NH 2 In Situ Formation of Imines with Small Molecule Organic Aldehydes in Alkaline Borate Buffer

[0177]

[0178] DNA-NH 2(The initial head fragment mentioned in our previous patent CN108070009A) was dissolved in 250mM, pH=9.4 boric acid buffer solution, prepared into a 1mM concentration solution, stored in a 0.5mL centrifuge tube (5μL, 5nmol), and added to the solution Small molecule organic aldehyde (3.0 μL, 600 nmol, 200 mM MeCN solution, 120 molar equivalents) was added, vortexed and mixed, and reacted at room temperature for 30 minutes.

[0179] 2...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to an alkaline buffering solution for establishing a DNA coding compound library and an application thereof. The alkaline buffering solution is a boric acid buffering solution with the concentration of 250mM, the electric conductivity of the boric acid buffering solution is 5177ppm+/-10 percent, the density is 1.0246 g/mL+/-3 percent, and the pH is 9.4+/-0.1. The buffering solution has the characteristics of wide substrate application range and high universality and is applied to the amide synthetic reaction of DNA-CO2H and small molecular organic amine and applied to thereduction ammonolysis reaction for generating imine from DNA-NH2 and small molecular organic aldehyde and reducing the imine by virtue of sodium borohydride, and has the advantage of high conversionrate. The various alkaline buffering solutions prepared in the invention and used for synthesizing the DNA coding compound library is mild in reaction conditions, simple in operation, wide in substrate application range, relatively high in applicability and suitable for the mass synthesis of the DNA coding compound library.

Description

technical field [0001] The invention belongs to the field of organic chemistry, and relates to preparing buffers with a certain pH value according to a certain formula, selecting these buffers to be used in the construction of a DNA-encoded compound library, and obtaining the reaction conditions under several alkaline buffers through conditional screening , can be applied to the condensation reaction of the carboxyl group on the DNA substrate and the small molecule organic amine, and the reductive amination reaction of the amino group on the DNA substrate and the small molecule organic aldehyde. The substrates of these on-DNA reaction conditions are suitable It has a wide range of properties, mild reaction conditions, short reaction time and strong applicability, and is suitable for large-scale application in the synthesis of DNA-encoded compound libraries. Background technique [0002] Sydney Brenner and Professor Richard Lerner of the Scripps Research Institute in the Unit...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C07H1/00C07H21/04
CPCC07H1/00C07H21/04
Inventor 吴阿亮张朝欣曲毅胡雨睛徐艳芬张在红李科裴增飞陈雯婷蒯乐天杨洪芳彭宣嘉
Owner 上海药明康德新药开发有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap