CN-122011071-A - Synthesis method and application of DNA-ketoamide conjugate

Abstract

The application relates to the technical field of DNA coding molecular libraries, and particularly discloses a synthesis method and application of a DNA-ketoamide conjugate. Wherein, adding an oxidant into the DNA-hydroxyamide conjugate solution with the general formula (I) in the specification, and reacting for at least 1h at the temperature of-5 ℃ to obtain the DNA-ketoamide conjugate with the general formula (II) in the specification. The synthesis method has the characteristics of high selectivity, mild reaction conditions, compatibility of biological systems and the like, effectively ensures that other sensitive groups in DNA molecules are not influenced in the reaction process, and has higher conversion rate.

Inventors

• LI YIZHOU
• Fang Xianfu
• WANG HUIHONG
• LI YANGFENG
• FAN XIAOHONG
• WANG XIN

Assignees

• 重庆大学

Dates

Publication Date

20260512

Application Date

20251211

Claims (10)

1. 1. A method for synthesizing a DNA-keto amide conjugate, comprising the steps of: adding an oxidant into a DNA-hydroxyamide conjugate solution shown in a general formula (I), and reacting at least 1h ℃ under the condition of-5 ℃ to obtain a DNA-ketoamide conjugate shown in a general formula (II); wherein the structural formula of the DNA-hydroxyamide conjugate shown in the general formula (I) is shown as follows: ; The structural formula of the DNA-ketoamide conjugate shown in the general formula (II) is shown as follows: ; wherein R 1 is selected from alkyl primary amine with 3-6 carbon atoms, branched chain alkyl primary amine with 4 carbon atoms, six-membered cycloalkyl primary amine, amino acid, substituted benzyl amine with substituent groups independent from each other selected from any one or more of alkyl, alkoxy, halogen, hydroxy, trifluoromethyl, amino, ester group, amide group, nitro, cyano or phenyl, aniline, asymmetric secondary amine with 5 carbon atoms and six-membered secondary amine; R 2 is selected from an aromatic benzene ring substituted by one or more substituents independently selected from hydrogen, halogen, trifluoromethyl, carboxyl, amino, nitro, cyano, hydroxyl, phenyl, ester and amide groups, or isopropyl, or alkyl with 1-4 carbon atoms.
2. 2. The method of synthesizing a DNA-keto amide conjugate according to claim 1 wherein the oxidizing agent is selected from at least one of potassium ruthenate, dimethyl phthalate, an amine oxide salt, 2, 6-tetramethylpiperidine oxide, ammonium tetrapropylpiperhenate, sodium periodate, iodobenzene acetate, 2-iodoxybenzoic acid, ruthenium trichloride, and iron oxyhydroxide.
3. 3. The method for synthesizing a DNA-ketoamide conjugate according to claim 1, wherein the ratio of the DNA-hydroxyl amide conjugate represented by the general formula (I) to the oxidant is 1:15-75 in terms of molar equivalent ratio.
4. 4. The method for synthesizing a DNA-ketoamide conjugate according to claim 1, wherein the oxidizing agent is dissolved in an alkali solution of 0.4-0.6M and then added to the DNA-hydroxyamide conjugate solution.
5. 5. The method for synthesizing a DNA-ketoamide conjugate according to claim 1, wherein the reaction time is 1 to 5 hours.
6. 6. The method of synthesizing a DNA-keto amide conjugate according to claim 1 wherein the reaction temperature is 0 ± 1 ℃.
7. 7. The method for synthesizing a DNA-ketoamide conjugate according to any one of claims 1 to 6, wherein in the structural formula of the general formula (I), the DNA comprises a single-or double-stranded nucleotide chain obtained by polymerizing a nucleotide monomer, the nucleotide chain being linked to an α -hydroxy acid through one or more chemical bonds or groups; And/or the number of the groups of groups, In the structural formula of the general formula (II), the DNA comprises a single-stranded or double-stranded nucleotide chain obtained by polymerizing nucleotide monomers, and the nucleotide chain is connected with alpha-keto acid through one or more chemical bonds or groups.
8. 8. The method for synthesizing a DNA-ketoamide conjugate according to claim 7, wherein the nucleotide chain is at least one selected from the group consisting of an artificially modified nucleotide chain and an unmodified nucleotide chain.
9. 9. The method of synthesizing a DNA-keto amide conjugate according to any of claims 1 to 6 further comprising the step of detecting the product using an ultra high performance liquid chromatography mass spectrometer.
10. 10. Use of a DNA-keto amide conjugate synthesized by the synthesis method of a DNA-keto amide conjugate according to any one of claims 1 to 9 as an antiviral drug ingredient.

Description

Synthesis method and application of DNA-ketoamide conjugate Technical Field The application relates to the technical field of DNA coding molecular libraries, in particular to a synthesis method and application of a DNA-ketoamide conjugate. Background The DNA coding molecule library combines a large number of chemical molecule blocks into a huge scale compound library by a combinatorial chemistry synthesis method. At the same time, each compound molecule has a unique DNA strand attached to it that acts as a "barcode" for the compound. By amplifying and sequencing the DNA sequence of the bound compound, the compound can be efficiently identified. The DNA-ketoamide conjugate is formed by constructing an alpha-ketoamide structure on a DNA molecule, and the alpha-ketoamide structure has unique geometric conformation, so that the molecule can be endowed with good pharmacokinetic property and metabolic stability, and the structure has nucleophilic and electrophilic reactivity. When the alpha-ketoamide structure is used as the electrophilic fragment, the target binding capacity can be improved through enhancing the hydrogen bond action, and when the alpha-ketoamide structure is used as the electrophilic fragment, the carbonyl can form a covalent bond with cysteine of a protease catalytic site, so that high-efficiency inhibition is realized. This property makes it widely applicable in the fields of antiviral drug design (such as coronavirus main protease inhibitor), antibacterial agents, immunomodulators, etc., typically represented by telaprevir, an anti-hepatitis c drug, and broad-spectrum antiviral compounds. The alpha-ketoamide structure provides a key chemical basis for developing high-activity and low-toxicity targeted drugs by regulating and controlling the rigidity and reaction specificity of molecules. This provides more structural options for the discovery of novel drug molecules. However, the DNA therein has been reported to be an effective synthesis method for DNA-keto amide conjugates because of the milder reaction conditions, high efficiency and wide substrate applicability, which limits the expansion of the reaction kit to some extent. Therefore, there is a need to develop methods for synthesizing DNA-keto amide conjugates. Disclosure of Invention The application aims to provide a synthesis method and application of a DNA-ketoamide conjugate, which are used for solving the problem that the DNA-ketoamide conjugate is not effectively synthesized at present. In order to achieve the object of the above application, in a first aspect, the present application provides a method for synthesizing a DNA-keto amide conjugate. The synthesis method of the DNA-ketoamide conjugate comprises the following steps: adding an oxidant into a DNA-hydroxyamide conjugate solution shown in a general formula (I), and reacting at least 1h ℃ under the condition of-5 ℃ to obtain a DNA-ketoamide conjugate shown in a general formula (II); Wherein, the structural formula of the DNA-hydroxyamide conjugate shown in the general formula (I) is shown as follows: ; The structural formula of the DNA-ketoamide conjugate shown in the general formula (II) is shown as follows: ; wherein R 1 is selected from alkyl primary amine with 3-6 carbon atoms, branched chain alkyl primary amine with 4 carbon atoms, six-membered cycloalkyl primary amine, amino acid, substituted benzyl amine with substituent groups independent from each other selected from any one or more of alkyl, alkoxy, halogen, hydroxy, trifluoromethyl, amino, ester group, amide group, nitro, cyano or phenyl, aniline, asymmetric secondary amine with 5 carbon atoms and six-membered secondary amine; R 2 is selected from an aromatic benzene ring substituted by one or more substituents independently selected from hydrogen, halogen, trifluoromethyl, carboxyl, amino, nitro, cyano, hydroxyl, phenyl, ester and amide groups, or isopropyl, or alkyl with 1-4 carbon atoms. In some embodiments, the molar equivalent of the oxidizing agent is 15-75. In some embodiments, the oxidizing agent is first dissolved in 0.4-0.6M lye and then added to the DNA-hydroxyamide conjugate solution. In some embodiments, the reaction time is 1h to 5 h. In some embodiments, the reaction temperature is 0 ± 1 ℃. In some embodiments, in the structural formula of formula (I) shown, the DNA comprises a single-or double-stranded nucleotide chain polymerized from nucleotide monomers, the nucleotide chain being linked to an α -hydroxy acid by one or more chemical bonds or groups; And/or the number of the groups of groups, In the structural formula of the general formula (II), the DNA comprises a single-stranded or double-stranded nucleotide chain obtained by polymerizing nucleotide monomers, and the nucleotide chain is connected with alpha-keto acid through one or more chemical bonds or groups. In some embodiments, the nucleotide chain is selected from at least one of an artificially modified nucleotide chain, an unmodified nuc