Preparation method of alkynyl-containing microspheres
Alkynyl-containing microspheres were prepared by grafting alkynyl functional groups on the polymer backbone, which solved the problem of lack of biomacromolecular enrichment methods in the prior art, and achieved effective alkynyl-based microsphere preparation and enrichment effects.
Patent Information
- Application Number
- CN202411814792.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-11
- Publication Date
- 2025-03-18
AI Technical Summary
In the prior art, biomacromolecules containing azide groups by metabolization lack effective means of enrichment.
Alkynyl-containing microspheres were prepared by grafting alkynyl functional groups by using a polymer backbone and propargyl glycidyl ether as raw materials.
The prepared alkynyl-containing microspheres can be effectively enriched, with an alkynyl content ranging from 100 to 150 μmol/g, providing an effective enrichment method.
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Figure CN119638869A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of chemical engineering, and in particular to a method for preparing alkyne-containing microspheres. Background Art
[0002] Click chemistry is one of the most "robust" chemical tools in modern chemical biology. It has been widely used in the fields of molecular glue, proximity labeling, metabolic labeling, etc., making invisible biomacromolecules "visible and tangible". However, there is often no good enrichment method for biomacromolecules containing azide groups through metabolic labeling. To meet this demand, this paper invented a microsphere containing alkyne groups.
[0003] References:
[0004] Sharpless, KBet al. Angew. Chem. Int. Ed. 2002, 41, 1053.
[0005] Sharpless,KBet al.J.Am.Chem.Soc.2005,127,6686. Summary of the invention
[0006] In order to solve the above problems, the present invention provides a method for preparing alkynyl-containing microspheres. The present invention selects a polymer skeleton with good biocompatibility and affinity and propargyl glycidyl ether as raw materials, and grafts the alkynyl functional group onto the polymer skeleton through a ring-opening reaction between a hydroxyl or amino group and an epoxy group, thereby obtaining a method for preparing alkynyl-containing microspheres.
[0007] In order to achieve the above object, the present invention provides the following technical solutions:
[0008] The present invention provides a method for preparing alkyne-containing microspheres, comprising the following steps:
[0009] 1) mixing the polymer skeleton with an organic solvent and subjecting the mixture to heat preservation treatment to obtain a heat preservation material;
[0010] The polymer skeleton contains hydroxyl groups or amino groups;
[0011] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50-110° C. for 7-20 hours.
[0012] Preferably, the polymer skeleton in step 1) comprises one or more of agarose microspheres, agarose primary amine microspheres and polyethylene primary amine resin.
[0013] Preferably, in step 1), the volume ratio of the mass of the polymer skeleton to the organic solvent is 20 g:100 mL;
[0014] The organic solvent includes acetone and / or toluene.
[0015] Preferably, the conditions for the heat preservation treatment in step 1) include: a temperature of 40 to 100° C. and a time of 1 to 2 hours.
[0016] Preferably, in step 2), the volume ratio of propargyl glycidyl ether to the organic solvent is 1:5.
[0017] Preferably, the temperature of mixing propargyl glycidyl ether and the organic solvent in step 2) is controlled to be 40°C.
[0018] Preferably, in step 1), the polymer skeleton is pre-treated and then mixed with an organic solvent, and the pre-treatment comprises the following steps: washing the polymer skeleton with deionized water, and then washing with an organic solvent or water;
[0019] The mass ratio of the polymer skeleton to deionized water is 1:5;
[0020] The mass ratio of the polymer skeleton to the volume of the organic solvent is 20 g:500 mL;
[0021] The mass ratio of the polymer skeleton to the volume of water is 20g:500mL, and the water is deionized water.
[0022] Preferably, when the polymer skeleton is agarose microspheres, the method for preparing the alkyne-containing microspheres comprises the following steps:
[0023] 1) mixing agarose microspheres and acetone, and heat-insulating to obtain an insulation material;
[0024] The mass ratio of the agarose microspheres to the volume of acetone is 20 g:100 mL;
[0025] The temperature of the heat preservation treatment is 40°C and the time is 2h;
[0026] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 20 hours;
[0027] The volume ratio of the propargyl glycidyl ether to acetone is 1:5;
[0028] The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.
[0029] Preferably, when the polymer skeleton is agarose primary amine microspheres, the method for preparing the alkyne-containing microspheres comprises the following steps:
[0030] 1) mixing agarose primary amine microspheres with acetone and heat-insulating the mixture to obtain an insulation material;
[0031] The mass ratio of the agarose primary amine microspheres to the volume of acetone is 20 g:100 mL;
[0032] The temperature of the heat preservation treatment is 40°C and the time is 0.5 to 24 hours;
[0033] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 7 to 12 hours;
[0034] The volume ratio of the propargyl glycidyl ether to acetone is 1:5;
[0035] The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.
[0036] Preferably, when the polymer skeleton is a polyethylene primary amine resin, the method for preparing the alkyne-containing microspheres comprises the following steps:
[0037] 1) mixing polyethylene primary amine resin and toluene, and heat-insulating the mixture to obtain a heat-insulating material;
[0038] The volume ratio of the polyethylene primary amine resin to toluene is 20 g:100 mL;
[0039] The temperature of the heat preservation treatment is 100°C and the time is 1h;
[0040] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 110° C. for 7 to 12 hours;
[0041] The volume ratio of the propargyl glycidyl ether to toluene is 1:5;
[0042] The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.
[0043] Beneficial effects of the present invention:
[0044] The alkynyl-containing microspheres prepared by the preparation method provided by the present invention have an alkynyl content of 100 to 150 μmol / g as measured by iodine titration. BRIEF DESCRIPTION OF THE DRAWINGS
[0045] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.
[0046] Figure 1 is the reaction equation of Example 1;
[0047] Figure 2 is the reaction equation of Example 2;
[0048] Figure 3 It is the reaction equation of Example 3. DETAILED DESCRIPTION
[0049] The present invention provides a method for preparing alkyne-containing microspheres, comprising the following steps:
[0050] 1) mixing the polymer skeleton with an organic solvent and subjecting the mixture to heat preservation treatment to obtain a heat preservation material;
[0051] The polymer skeleton contains hydroxyl groups or amino groups;
[0052] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50-110° C. for 7-20 hours.
[0053] The present invention mixes a polymer skeleton with an organic solvent and performs heat preservation treatment to obtain an insulation material; the polymer skeleton contains hydroxyl groups or amino groups. In the present invention, the polymer skeleton preferably includes one or more of agarose microspheres, agarose primary amine microspheres and polyethylene primary amine resins. In the present invention, the agarose microspheres are derived from 6FF agarose microspheres sold by Ningbo Zhengguang Resin Co., Ltd., and the 6FF agarose microspheres have hydroxyl groups. In the present invention, the agarose primary amine microspheres are derived from Ningbo Zhengguang Resin Co., Ltd., and the agarose primary amine microspheres have amino groups. In the present invention, the polyethylene primary amine resin is derived from Ningbo Zhengguang Resin Co., Ltd., and the polyethylene primary amine resin has amino groups. In the present invention, the mass of the polymer skeleton and the volume ratio of the organic solvent are preferably 20g:100mL. In the present invention, the organic solvent preferably includes acetone and / or toluene. In the present invention, the conditions for the heat preservation treatment preferably include: a temperature of 40 to 100°C and a time of 1 to 2h. In the present invention, the polymer skeleton is preferably mixed with an organic solvent after pre-treatment, and the pre-treatment preferably includes the following steps: washing the polymer skeleton with deionized water, and then washing with an organic solvent or water. In the present invention, the mass ratio of the polymer skeleton to deionized water is preferably 1:5. In the present invention, the volume ratio of the mass of the polymer skeleton to the organic solvent is preferably 20g:500mL. In the present invention, the volume ratio of the mass of the polymer skeleton to water is preferably 20g:500mL, and the water is preferably deionized water. In the present invention, the purpose of the pre-treatment is to remove impurities from the polymer skeleton.
[0054] The present invention mixes the obtained insulation with propargyl glycidyl ether and reacts at 50-110° C. for 7-20 hours. In the present invention, the volume ratio of the propargyl glycidyl ether to the organic solvent is preferably 1:5. In the present invention, the temperature at which the propargyl glycidyl ether and the organic solvent are mixed is preferably controlled to be 40° C.
[0055] In the present invention, when the polymer skeleton is preferably agarose microspheres, the method for preparing the alkyne-containing microspheres preferably comprises the following steps:
[0056] 1) mixing agarose microspheres with acetone and heat-insulating to obtain an insulation material; the mass ratio of the agarose microspheres to the volume of acetone is 20 g:100 mL; the temperature of the heat-insulating treatment is 40° C. and the time is 2 h;
[0057] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 20 hours; the volume ratio of the propargyl glycidyl ether to acetone is 1:5; and the temperature of the propargyl glycidyl ether mixing is controlled to be 40° C.
[0058] In the present invention, when the polymer skeleton is preferably agarose-based primary amine microspheres, the preparation method of the alkyne-containing microspheres preferably comprises the following steps:
[0059] 1) mixing agarose primary amine microspheres with acetone and heat-insulating to obtain an insulation material; the mass ratio of the agarose primary amine microspheres to the volume of acetone is 20 g:100 mL; the temperature of the heat-insulating treatment is 40° C., and the time is 0.5-24 h;
[0060] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 7 to 12 hours; the volume ratio of the propargyl glycidyl ether to acetone is 1:5; and the temperature of the propargyl glycidyl ether mixing is controlled to be 40° C.
[0061] In the present invention, when the polymer skeleton is preferably a polyethylene primary amine resin, the preparation method of the alkyne-containing microspheres comprises the following steps:
[0062] 1) Mixing polyethylene primary amine resin and toluene, and heat-insulating treatment to obtain an insulation material; the mass ratio of the polyethylene primary amine resin to the volume of toluene is 20g:100mL; the temperature of the heat-insulating treatment is 100°C, and the time is 1h;
[0063] 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 110° C. for 7 to 12 hours; the volume ratio of the propargyl glycidyl ether to toluene is 1:5; and the temperature of the propargyl glycidyl ether mixing is controlled to be 40° C.
[0064] In order to further illustrate the present invention, the present invention is described in detail below in conjunction with embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0065] Example 1
[0066] Take 20g of 6FF agarose microspheres (Zhengguang brand) in an exchange column (20mm×100mm), use 100mL of deionized water to wash the 6FF agarose microspheres, and remove the impurities such as ethanol, water and inorganic salts contained in the agarose preservation solution of the 6FF agarose microspheres. Add 500mL of acetone to the exchange column to elute the 6FF agarose microspheres. Then transfer it to the reactor and add 100mL of acetone to enable it to be stirred in the reactor. Heat to 40℃ and keep warm for 2h. Keep the internal temperature at 40℃ and add 20mL of propargyl glycidyl ether. React at 50℃ for 20h. Filter out the agarose microspheres, wash the acetone with deionized water, and you can get agarose microspheres with alkynyl groups.
[0067] The alkynyl content was determined using the iodine titration method: 150 μmol / g.
[0068] Example 2
[0069] Take 20g of agarose primary amine microspheres (Zhengguang brand) and put them in an exchange column (20mm×100mm). Use deionized water to remove the water and impurities such as inorganic salts inside. Add 500mL of deionized water to the exchange column to wash the agarose primary amine microspheres. Then transfer it to a reactor and add 100mL of acetone to allow it to stir in the reactor. Keep the temperature at 40℃ for not less than 10s. Keep the internal temperature at 40℃ and add 20mL of propargyl glycidyl ether. React at 50℃ for 25.0h. Filter out the agarose microspheres, wash the acetone with deionized water, and you can get agarose microspheres with alkynyl groups.
[0070] The alkynyl content was determined by iodine titration: 120 μmol / g.
[0071] Example 3
[0072] Take 20g of styrene primary amine resin (Zhengguang brand) in an exchange column (20mm x 100mm), and use deionized water to remove impurities such as water and inorganic salts inside it. Add 500mL of toluene to the exchange column to wash the styrene primary amine resin. Then transfer it to a reactor and add 100mL of toluene to stir it in the reactor. Keep it at 100℃ for 1h. Keep the internal temperature at 40℃ and add 20mL of propargyl glycidyl ether. Then react at 110℃ for 7-12h. Filter out the agarose microspheres, wash the organic solvent with deionized water, and you can get agarose microspheres with alkynyl groups.
[0073] The alkynyl content was determined using the iodine titration method: 100 μmol / g.
[0074] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.
Claims
1. A method for preparing alkyne-containing microspheres, characterized in that: The following steps are involved: 1) mixing the polymer skeleton with an organic solvent and subjecting the mixture to heat preservation treatment to obtain a heat preservation material; The polymer skeleton contains hydroxyl groups or amino groups; 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50-110° C. for 7-20 hours.
2. The preparation method according to claim 1, characterized in that: The polymer skeleton in step 1) comprises one or more of agarose microspheres, agarose primary amine microspheres and polyethylene primary amine resins.
3. The preparation method according to claim 1, characterized in that: In the step 1), the volume ratio of the mass of the polymer skeleton to the organic solvent is 20 g:100 mL; The organic solvent includes acetone and / or toluene.
4. The preparation method according to claim 1, characterized in that: The conditions for the heat preservation treatment in step 1) include: a temperature of 40 to 100° C. and a time of 1 to 2 hours.
5. The preparation method according to claim 1, characterized in that: In the step 2), the volume ratio of propargyl glycidyl ether to the organic solvent is 1:
5.
6. The preparation method according to claim 1, characterized in that: In the step 2), the temperature of mixing propargyl glycidyl ether and the organic solvent is controlled to be 40°C.
7. The preparation method according to claim 1, characterized in that: In the step 1), the polymer skeleton is pre-treated and then mixed with an organic solvent, and the pre-treatment includes the following steps: Use deionized water to clean the polymer skeleton, and then use an organic solvent or water to clean it; The mass ratio of the polymer skeleton to deionized water is 1:5; The mass ratio of the polymer skeleton to the volume of the organic solvent is 20 g:500 mL; The mass ratio of the polymer skeleton to the volume of water is 20g:500mL, and the water is deionized water.
8. The preparation method according to claim 1, 2 or 7, characterized in that: When the polymer skeleton is agarose microspheres, the preparation method of the alkyne-containing microspheres comprises the following steps: 1) mixing agarose microspheres and acetone, and heat-insulating to obtain an insulation material; The mass ratio of the agarose microspheres to the volume of acetone is 20 g:100 mL; The temperature of the heat preservation treatment is 40°C and the time is 2h; 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 20 hours; The volume ratio of the propargyl glycidyl ether to acetone is 1:5; The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.
9. The preparation method according to claim 1, 2 or 7, characterized in that: When the polymer skeleton is agarose primary amine microspheres, the preparation method of the alkyne-containing microspheres comprises the following steps: 1) mixing agarose primary amine microspheres with acetone and heat-insulating the mixture to obtain an insulation material; The mass ratio of the agarose primary amine microspheres to the volume of acetone is 20 g:100 mL; The temperature of the heat preservation treatment is 40°C and the time is 0.5 to 24 hours; 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 50° C. for 7 to 12 hours; The volume ratio of the propargyl glycidyl ether to acetone is 1:5; The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.
10. The preparation method according to claim 1, 2 or 7, characterized in that: When the polymer skeleton is a polyethylene primary amine resin, the preparation method of the alkyne-containing microspheres comprises the following steps: 1) mixing polyethylene primary amine resin and toluene, and heat-insulating the mixture to obtain a heat-insulating material; The volume ratio of the polyethylene primary amine resin to toluene is 20 g:100 mL; The temperature of the heat preservation treatment is 100°C and the time is 1h; 2) mixing the insulation obtained in step 1) with propargyl glycidyl ether, and reacting at 110° C. for 7 to 12 hours; The volume ratio of the propargyl glycidyl ether to toluene is 1:5; The temperature of the propargyl glycidyl ether mixture was controlled to be 40°C.