Connecting arm of immobilized nucleic acid aptamer as well as preparation method and application of the connecting arm

A nucleic acid aptamer and linker technology, which is used in the preparation of nucleic acid aptamer-drug conjugates. The linker field of nucleic acid aptamers can solve the problem of low selectivity and difficulty in obtaining linker raw materials, etc. problem, to achieve high selectivity, less side reactions, and easy to obtain effects

Pending Publication Date: 2020-04-21
GANNAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, this immobilized nucleic acid aptamer linker has the following problems: 1) the selectivity is not high when using concentrated ammonia water to excise CPG in the last step of nucleic acid aptamer synthesis, and you may get such figure 2 The by-product shown in a; 2) a primary alcohol is retained on the resulting nucleic acid aptamer linking arm, such as figure 2 As shown in b, this primary hydroxyl group is easy to transesterify with the 2-hydroxyl phosphate; 3) the raw materials for preparing the connecting arm are not easy to obtain

Method used

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  • Connecting arm of immobilized nucleic acid aptamer as well as preparation method and application of the connecting arm
  • Connecting arm of immobilized nucleic acid aptamer as well as preparation method and application of the connecting arm
  • Connecting arm of immobilized nucleic acid aptamer as well as preparation method and application of the connecting arm

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Embodiment approach

[0044] According to the present invention, the amino protecting group can be any group known in the art that can protect the amino to avoid chemical reaction. According to a preferred embodiment of the present invention, the amino protecting group is a halogen Substituted acetyl.

[0045] Wherein, the halogen may be chlorine or fluorine, preferably fluorine.

[0046] According to the present invention, the number of halogen substituents can be 1 or more, preferably 3.

[0047] According to a most preferred embodiment of the present invention, the amino protecting group is a trifluoroacetyl group.

[0048] According to the present invention, n can be any integer of 2, 3, 4, 5, 6, preferably, n is 2, 3 or 4.

[0049] In a second aspect, the present invention provides a method for preparing a tether of an immobilized nucleic acid aptamer, the method comprising:

[0050] (1) In the presence of the first catalyst, the nitrile of acrylonitrile or the substituted C3-C7 of the ω-ea...

Embodiment 1

[0149] This example is used to illustrate the synthesis of the tether of the nucleic acid aptamer immobilized provided by the present invention

[0150] (1) Synthesis of the first intermediate product

[0151]

[0152] Suspend 0.8g of sodium hydride (60% by weight, 20mmol) in 250mL of anhydrous tetrahydrofuran, stir in an ice-water bath, add 13.2g of compound 1 acetonide (100mmol) dropwise, after the addition is complete, gradually warm up to room temperature, and stir for 2 hours , add 13.2mL acrylonitrile (200mmol) dropwise under cooling in an ice-water bath, after the dropwise addition is complete, stir at room temperature for 2 hours, add 50mL of water, distill off THF under reduced pressure, extract with dichloromethane, collect the oil phase, concentrate, and distill under reduced pressure to obtain 16.8 g of oil (90.9% yield). 1 H NMR (400MHz, CDCl 3 ): δ1.35(s,3H),1.41(s,3H),2.61(t, J=6.3Hz,2H),3.51-3.58(m,2H),3.69-3.76(m,3H),4.05( dd, J=8.2Hz, J=6.4Hz, 1H), 4.23...

Embodiment 2

[0169] This example is used to illustrate the synthesis of the tether of the nucleic acid aptamer immobilized provided by the present invention

[0170] The synthesis of the linker arm of the immobilized nucleic acid aptamer was carried out according to the method in Example 1, except that in step (1), acrylonitrile was replaced with 4-bromobutyronitrile. The ammonolysis of a part of the product was taken and detected, which proved that the tether of the nucleic acid aptamer immobilized in the present invention was successfully obtained, and the yields of each step are shown in Table 1.

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Abstract

The invention relates to the technical field of aptamers, and discloses a connecting arm of an immobilized nucleic acid aptamer as well as a preparation method and application of the connecting arm. The connecting arm has a structure as shown in a formula 1 shown in the specification, wherein R1 and R2 are independently selected from straight-chain or branched-chain C1-C10 alkyl groups, R is an amino protection group, and n is an integer from 2 to 6. Compared with the prior art, the connecting arm has the following beneficial effects: 1) the connecting arm of the immobilized nucleic acid aptamer provided by the invention has high selectivity when CPG is excised by using concentrated ammonia water in the last step of synthesizing the aptamer, and only one product can be obtained; 2) only one secondary alcohol is left on the connecting arm of the nucleic acid aptamer, and side reaction is not liable to occur, and 3) the raw material for preparing the connecting arm of the immobilized nucleic acid aptamer is solketal and is easy to obtain.

Description

technical field [0001] The present invention relates to the technical field of nucleic acid aptamers, in particular to a connecting arm of an immobilized nucleic acid aptamer, a preparation method for a connecting arm of an immobilized nucleic acid aptamer and an immobilized aptamer prepared by the method. The connecting arm of the nucleic acid aptamer, and the application of the connecting arm of the nucleic acid aptamer immobilized in the preparation of the nucleic acid aptamer-drug conjugate. Background technique [0002] Nucleic acid aptamer is a single-stranded oligonucleotide that binds to a specific target by using its self-folded higher-level structure, and can be screened by systematic evolution of ligands by exponential enrichment (SELEX). Nucleic acid aptamers have similar affinity and specificity to antibodies, and are less toxic and safer for clinical application [Wochner, A.; Menger, M.; Rimmele, M. Characterization of aptamers for therapeutic studies. Expert ...

Claims

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Application Information

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IPC IPC(8): C07C231/02C07C233/18C12N15/115A61K47/54
CPCC07C233/18C12N15/115A61K47/549C12N2310/16C12N2310/33Y02P20/55
Inventor 江峰谢欣易小庆陈伟铭陈垒钟佳宁方小军郭宇许小玲彭维杰
Owner GANNAN MEDICAL UNIV
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