Method for preparing DNA coding compound library, initial head fragment compound and prepared DNA coding compound

A compound and fragment technology, applied in the field of combinatorial chemistry, can solve problems such as tedious coding transformation, heavy workload, and difficult screening

Active Publication Date: 2018-05-25
上海药明康德新药开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, with the further research, people found that single-target drugs also have obvious shortcomings: first, most diseases are not caused by a single gene or target, and diseases caused by abnormal single genes or targets account for only a small number; Drug targets are not unique to a certain disease. Most of them have multiple biological functions. Excessively inhibiting or activating a certain biomolecule in the body may affect the function of related biomolecules while interfering with its own biological functions, resulting in Side effects; third, it is difficult to find molecules that only act on a single target
However, this method also has great limitations. First, there are few cycles, and the number of compound molecules obtained is limited. Generally, it is difficult to reach the million level. Compared with the DNA Recording Molecule Library (DRCL), there a

Method used

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  • Method for preparing DNA coding compound library, initial head fragment compound and prepared DNA coding compound
  • Method for preparing DNA coding compound library, initial head fragment compound and prepared DNA coding compound
  • Method for preparing DNA coding compound library, initial head fragment compound and prepared DNA coding compound

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1, the preparation of starting head fragment (SO-1)

[0075] A 5' phosphorylated nucleotide chain with the following sequence was synthesized as a starting head fragment and purified by HPLC by Suzhou Jinweizhi Biotechnology Co., Ltd.

[0076] 5'-PO 4 2- CTGCATA AGGCCT TAGTCTTdTTTdTTTGACTA AGGCCT TATGCAGCC-3' (SEQ ID NO: 01, MW: 14460.2, Figure 11)

[0077] dT has the following structure:

[0078]

[0079] The base was obtained by using the following monomer in a DNA synthesizer: Fmoc Amino-Modifier C6-dT-CEPhosphoramidite (MFCD08061991, CAS: 210534-16-0).

[0080] Part of the bases at both ends of the nucleotide chain is complementary pairing, which can form a large hairpin structure with a protruding base sequence CC and two cleavage sites ( AGGCCT ), the cleavage site for the nicking variant of the restriction endonuclease StuI. There are two bases in the middle of the hairpin ring, and there is a 6-carbon alkyl chain at the side chain C5 of t...

Embodiment 2

[0082] Embodiment 2, the preparation of the complete head fragment (SO-2) that has linker

[0083] The initial head fragment obtained in Example 1 was reacted with Fmoc-PEG5-NHS ester and Acid-PEG5-NHS ester to prepare the head fragment SO-2 with the following sequence.

[0084] 5'-PO 4 2- CTGCATA AGGCCT TAGTCTTdT(X)TTdT(Y)TTGACTA AGGCCT TATGCA GCC-3' (sequence number: 02, molecular weight: 15294.2)

[0085] X and Y represent any one of the following structures:

[0086]

[0087]

[0088]100 nmol of the starting head fragment (SO-1) was dissolved in 100 μl of sodium borate buffer solution (pH=9.5, 250 mM), 200 nmol of Fmoc-PEG5-NHS ester (MFCD28976702, CAS: 1402080- 11-8, molecular weight: 628.67, customized by Shanghai WuXi Kangde New Drug Development Co., Ltd.) was dissolved in 20uL dimethylacetamide (DMAc) and reacted in the solution of SO-1 at room temperature. Add 12 microliters of 5N sodium chloride solution and 360 microliters of cold ethanol to the reacti...

Embodiment 3

[0090] Example 3, Preparation of a DNA-encoded compound library with two pharmacophores in a 10×10 library

[0091] Step 1: Ligation of Initiating Primer to Complete Head Fragment SO-2 to Get Primed Head Fragment SO-2-P

[0092] In this exemplary DNA-encoded compound library, a fixed-coded nucleotide duplex (referred to as the initial primer, customized by Suzhou Jinweizhi Biotechnology Co., Ltd., HPLC purified) was connected to SO-2 for subsequent screening. General primer fragments for PCR of DNA-encoded compounds:

[0093] Starting primer upper chain: 5'-PO 4 2- -AAATCGATGACACAG-3' (SEQ ID NO: 03)

[0094] Starting primer lower strand: 5'-PO 4 2- -CATCGATTTGG-3' (sequence number: 04)

[0095] Mix 100 microliters of 100 nanomolar complete head fragment SO-2 aqueous solution with 110 microliters of a mixed aqueous solution of 110 nanomolar initial primer upper chain and 110 nanomolar initial primer lower chain, and then place in a PCR machine for 90 ℃ heating for 5 min...

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Abstract

The invention discloses a method for preparing a DNA coding compound library containing two or more pharmacophores. A novel initial head fragment used by the method has two or more chemically-modifiedlinking groups and comprises a nucleotide chain, 5' and 3' of the nucleotide chain respectively have a group R1 and a group R2 capable of performing chemical reaction, n connecting heads are arrangedon the nucleotide chain, each of the connecting heads is a long chain extended after a nucleotide monomer on the nucleotide chain is modified, and the long chain has a site G capable of performing chemical reaction. The site G of the initial head fragment respectively reacts with a fragment compound to generate a fragment compound residue B connecting the site G with the fragment compound together, a DNA coding compound is formed, and finally, the DNA coding compound library containing the two or more pharmacophores can be obtained. Through the method disclosed by the invention and the initial head fragment, the DNA coding compound library containing the two or more pharmacophores can be efficiently, simply and quickly obtained.

Description

technical field [0001] The present invention relates to the field of combinatorial chemistry, in particular to a method for preparing a library of DNA-encoded compounds containing two or more pharmacophore, the prepared DNA-encoded compound and the initial head segment compound. This method adopts a novel starting head fragment for the construction of a DNA-encoded compound library. The starting head fragment has two or more linking groups that can be chemically modified. A DNA-encoded compound library of one or more pharmacophores. Background technique [0002] A drug target refers to a biological molecule that has a causal relationship with the occurrence of a certain disease or participates in the development of the disease, and achieves the purpose of treating the disease by regulating its activity through a certain type of drug. Since the introduction of in vitro screening (invitro) Since its conception, with the advancement of molecular biology, proteomics, genomics a...

Claims

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Application Information

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IPC IPC(8): C07H1/00C07H21/04C12P19/34C40B40/06C40B50/06
CPCC07H1/00C07H21/04C12P19/34C40B40/06C40B50/06Y02P20/55
Inventor 吴阿亮张在红裴增飞黄书宽龚平秀周园园李科陈雯婷邢莉杨洪芳彭宣嘉
Owner 上海药明康德新药开发有限公司
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