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Method and kit for synthesizing and screening lead compound

A technology for lead compounds and screening methods is applied in the field of lead compound synthesis and screening methods and kits, and can solve the problems of large errors in double-stranded DNA sequencing results, difficulty in synthesizing target compounds, poor diversity of compound libraries, and the like. The effect of connection, stable structure and large storage capacity

Active Publication Date: 2014-06-25
HITGEN INC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, when double-stranded DNA is used to mark synthetic building blocks or compounds, the double-stranded DNA is easily cross-linked during the ligation extension process to form a coiled tertiary structure. During sequencing, it needs to be melted, and the operation is more complicated. Double-stranded DNA is used to mark When there are more than three steps of linear combination reactions, the sequencing results of double-stranded DNA have large errors, so that the method can only stay in two dimensions, so the number of compounds in the library can only be increased by increasing the number of synthetic building blocks in each step of the reaction. The obtained compound library has poor diversity, and it is difficult to synthesize the target compound

Method used

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  • Method and kit for synthesizing and screening lead compound
  • Method and kit for synthesizing and screening lead compound
  • Method and kit for synthesizing and screening lead compound

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1 Synthesis and screening method of lead compound of the present invention

[0060] 1. Preparation method

[0061] (1) Take synthetic building blocks and single-stranded DNA fragments:

[0062] i kinds of synthetic building blocks and (i+2) kinds of single-stranded DNA fragments, (i+2) kinds of single-stranded DNA fragments include i kinds of marker sequences, a kind of start sequence and a kind of end sequence, and the i kinds of marker sequences are specifically labeled i kinds of synthetic blocks, where i=1, 2, 3...n;

[0063] Cytidine can be linked to the following marker sequences to improve the ligation efficiency of subsequent single-stranded DNA fragments.

[0064] Table 1 Single-stranded DNA fragments

[0065]

[0066]

[0067]

[0068] (2) Synthesis: such as figure 1 As shown, the compound library was synthesized by combinatorial chemistry:

[0069] a. Prepare the initial synthetic building blocks: select 1~i synthetic building block...

Embodiment 2

[0092] Embodiment 2 uses the method of the present invention to form and screen the lead compound of trypsin inhibitor

[0093] 1. Materials and reagents

[0094] T4PNK(500U NEB-M0201V), T4 RNA ligase1(NEB-M0204S), Cartridges(PCR purification Kit(cat.no 28104,Nucleotides removal Kit cat.no 28306) purchased from Qiagen(Hilden,Germany).dNTPs(0.5mM, NEB, cat. no89009).

[0095] The single-stranded DNA fragments shown in Table 1 were synthesized by Genscript and Biosune.

[0096] 2. Preparation method

[0097] (1) Preparation of single-stranded DNA fragments:

[0098] 55 types of synthetic building blocks and 57 types of single-stranded DNA fragments, 57 types of single-stranded DNA fragments include 55 types of marker sequences, 1 type of start sequence and 1 type of end sequence;

[0099] Cytidine can be linked to the following marker sequences to improve the ligation efficiency of subsequent single-stranded DNA fragments.

[0100] Table 2 Single-stranded DNA fragments

[...

Embodiment 3

[0183] Example 3 The synthesis and screening kit of the lead compound of the present invention

[0184] 1. The composition of the kit of the present invention (the synthetic dosage of N synthetic blocks)

[0185] 1) i kinds of synthetic building blocks and i+2 kinds of single-stranded DNA fragments, the single-stranded DNA fragments are divided into start sequence, end sequence and i kinds of marker sequences, and the i kinds of marker sequences are the codes of i kinds of synthetic building blocks, among which , i=1, 2, 3...n;

[0186] Table 3 Synthetic building blocks and single-stranded DNA fragments of different sequences

[0187]

[0188]

[0189]

[0190] 2) Reagents for linking the initial sequence and synthetic building blocks, reagents for synthesis by combinatorial chemistry methods, and reagents for linking single-stranded DNA fragments;

[0191] Table 4 Reagents for linking the starting sequence and the synthetic building block

[0192] Reagent...

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Abstract

The invention discloses a method for synthesizing and screening a lead compound. The method comprises the following steps of (1) taking raw materials: taking i synthetic building blocks and (i+2) single-chain DNA segments; (2) synthesizing with a combined chemistry method to obtain a single-chain DNA marked compound library, wherein initial synthetic building blocks are marked by fluorescent molecules; (3) screening: screening the compound DNA marked compound library; and (4) sequencing: taking the DNA marked compound obtained by screening of the step (3), sequencing DNA on the DNA marked compound, and determining the building block and reaction process of the compound according to the DNA sequence. The invention also discloses a kit for synthesizing and screening the lead compound, and a combined chemistry library. The method and the kit provided by the invention can synthesize and screen to obtain a target lead compound rapidly and effectively, and simultaneously, monitor the reaction process. Operations are simple and cost is low.

Description

technical field [0001] The invention relates to the field of chemistry, in particular to a synthesis and screening method and kit of a lead compound. Background technique [0002] Since the late 1980s, with the breakthrough of molecular biology research - the development of high-throughput screening technology, the number of new molecular entities required for the development of new drugs has increased, and scientists have shifted their attention from searching for natural products to synthetic large ones. A large number of compound groups - chemical library, chemical library is composed of many organic compounds with different properties. Combinatorial chemical synthesis is a technique for synthesizing chemical libraries. Using this technique, different series of synthetic building blocks—reactive components—are arranged in an orderly manner to form a large series of diverse molecular entity groups. Combinatorial chemical synthesis is often referred to as a number game, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C40B40/04C40B50/04C40B20/04C40B70/00
CPCC40B70/00C40B40/04C12N15/1068C40B20/04C12N15/1093C40B50/04C40B50/10
Inventor 李进窦登峰陈仰王星江洋陈湜瞿庆喜万金桥钟丽娜葛啸虎黄奇
Owner HITGEN INC