Erythromycin chiral silica gel filler and preparation method thereof
By bonding erythromycin molecules to γ-GPS and combining with silica gel, an erythromycin chiral silica filler was prepared, which solved the problem of unstability of existing macrocyclic glycopeptide chiral stationary phases under extreme conditions, achieved efficient separation of specific enantiomers in complex samples and stability of fillers, reducing the preparation cost.
Patent Information
- Application Number
- CN202411977129.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2025-03-28
AI Technical Summary
The existing macrocyclic glycopeptide chiral stationary phases are unstable under extreme pH or temperature conditions, and the separation effect on certain specific structures is not ideal, and the column effect and column life are affected by the use conditions.
A erythromycin chiral silica filler was prepared by bonding erythromycin molecules to gamma-epoxypropoxypropyltrimethoxysilane (γ-GPS) and combining with silica gel, and end-group tail sealing treatment was performed to improve the stability of the filler.
Effective separation of specific enantiomers in complex samples is achieved, the filler is stable within the pH range of 2-8, can withstand a variety of solvents, extending the long life and stability of the column, and reducing the preparation cost.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field related to chiral chromatographic fillers, in particular to an erythromycin chiral silica gel filler and a preparation method thereof. Background Art
[0002] Chirality is widely present in nature and is a basic characteristic of materials. Enantiomers usually have different biological properties and have completely different effects on organisms. Therefore, the separation of enantiomers is necessary in the pharmaceutical field. Since Pasteur first successfully separated racemic tartrate with tweezers under a microscope in 1848, chiral separation technology has developed rapidly. As far as we know, mechanical, chemical, biochemical, chromatographic, extraction and membrane separation techniques have been explored for chiral separation. Chromatographic separation methods, especially HPLC technology combined with chiral fixation, are considered to be one of the most convenient and effective methods among the various chiral drug separation technologies currently available.
[0003] Since the introduction of the first chiral stationary phase (CSP) in 1938, the planning and development of new CSPs for HPLC has been considered a continuous and evolving issue. HPLC using chiral stationary phases has proven to be a very useful and accurate analytical tool and is widely used in different fields and professions. CSPs are the key to determining chiral separation capabilities. The first requirement for a CSP to be applied to the separation of chiral compounds is that it can quickly and accurately determine the enantiomeric purity of different structures. In the past few decades, many different classes of chiral stationary phases have been widely reported as separation materials, of which more than 100 are currently on the market. The most commonly used chiral selectors are amino acids (natural or derived), proteins, cyclodextrins (natural or derived), derived linear or branched carbohydrates (such as cellulose or amylose) and cavity type selectors, such as macrocyclic antibiotics.
[0004] Macrocyclic antibiotics have a large number of chiral centers and characteristic groups, which can act as chiral devices to interact with analytes, such as electrostatic interactions. The types of macrocyclic antibiotics include ansamycins, glycopeptides, and polypeptides. Among these major types of antibiotics, glycopeptide antibiotics have the highest enantiomeric selectivity and are relatively stable during the bonding and column loading of the stationary phase. Therefore, the research on chiral stationary phases of macrocyclic antibiotics mainly focuses on glycopeptide antibiotics.
[0005] Although macrocyclic glycopeptide chiral stationary phases show good chiral selectivity for many compounds and can separate a variety of chiral compounds, they may not provide ideal separation effects for certain compounds with specific structures and may be unstable under extreme pH or temperature conditions. In practical applications, the column efficiency and column life of macrocyclic glycopeptide chiral stationary phases may be affected by the conditions of use. In order to solve the limitations of existing stationary phases, it is necessary to continuously research and develop new macrocyclic glycopeptide chiral stationary phases to meet the needs of different chiral separations.
[0006] Erythromycin is a widely used glycopeptide macrocyclic antibiotic with significant antibacterial activity. The erythromycin molecule contains multiple chiral carbon atoms, which determines that it has multiple stereoisomers. Erythromycin A is its main active ingredient, which contains 14 chiral centers, has two 6-membered rings and one macrocycle, and has 5 hydroxyl groups. It can produce multiple forces such as hydrophobic interaction, polar interaction, hydrogen bonding and π-π conjugation with the compounds to be separated. It has special stereoselectivity and can achieve effective separation of specific enantiomers in complex samples. It is an effective supplement to the separation ability and application range of existing chiral fillers. Summary of the invention
[0007] The present invention aims to provide an erythromycin chiral silica gel filler and a preparation process thereof. The present invention provides an erythromycin chiral silica gel filler, comprising: silica gel and γ-glycidoxypropyltrimethoxysilane (γ-GPS) bonded to the surface of the silica gel and erythromycin molecules. Wherein: the chemical structure of γ-GPS is:
[0008]
[0009] The chemical structure of erythromycin is:
[0010]
[0011] The present invention also provides a method for preparing the erythromycin chiral silica gel filler, comprising the following steps:
[0012] Step 1: preparing glycidoxylated silica gel: dissolving γ-glycidoxypropyltrimethoxysilane (γ-GPS) in a first solvent, adding silica gel, heating for reaction, cooling, filtering, washing, and drying to obtain glycidoxylated silica gel;
[0013] Step 2: Prepare erythromycin chiral silica gel filler: stir the second solvent and erythromycin, then add the glycidoxylated silica gel, heat to react, cool, add trimethylsilyl chloride to perform end-capping treatment, seal the remaining hydroxyl groups on the surface of the silica gel, filter, wash the filtered solid with water and methanol respectively, dry the filtered solid to obtain erythromycin chiral silica gel filler.
[0014] Furthermore, in step 1, the first solvent is methanol and / or ethanol, and the silica gel is spherical porous silica gel.
[0015] Furthermore, in step 1, the reaction temperature is 60-80° C., the reaction pressure is normal pressure, the reaction time is 4-28 hours, and inert gas protection is required during the reaction.
[0016] Furthermore, in step one, the mass ratio of silica gel to γ-glycidoxypropyltrimethoxysilane (γ-GPS) is 1:0.5-5, and the mass volume ratio of γ-glycidoxypropyltrimethoxysilane (γ-GPS) to the first solvent is 1:5-20.
[0017] Furthermore, in step 2, the second solvent is pyridine and / or DMSO.
[0018] Furthermore, the second solvent used in step 2 is an anhydrous solvent.
[0019] Furthermore, in step 2, the reaction temperature is 60-120° C., and the reaction time is 10-16 hours.
[0020] Furthermore, in step 2, the mass volume ratio of erythromycin to the second solvent is 1:5-15, the mass ratio of erythromycin to glycidoxylated silica gel is 1:0.1-2; and the mass ratio of erythromycin to trimethylchlorosilane is 1:1-5.
[0021] The beneficial effects of the present invention are as follows: 1. The present invention adopts γ-glycidoxypropyltrimethoxysilane as a spacer arm to bond erythromycin to macroporous silica gel, thereby preparing a novel chiral filler, which can realize effective separation of specific enantiomers in complex samples; 2. After the bonding reaction is completed, the present invention performs end-capping treatment on the filler to reduce nonspecific adsorption and improve the separation performance and stability of the filler; 3. The erythromycin chiral silica gel provided by the present invention has a wide range of applications, and its unique chemical structure is also suitable for separation applications in reverse phase and HILIC modes; 4. The erythromycin production process used in the present invention is mature and low in cost, which greatly reduces the cost of preparing chromatographic fillers; 5. The erythromycin chiral silica gel filler of the present invention can effectively separate enantiomers, and the filler is stable in the pH range of 2-8, can tolerate a variety of solvents, will not dissolve or structural damage, and ensures the long life and stability of the chromatographic column. DETAILED DESCRIPTION
[0022] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0023] Example 1
[0024] 1. Preparation of glycidoxylated silica gel:
[0025] In a reaction bottle, add 0.5g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 5ml of methanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 60°C, keep warm for 28 hours, cool to room temperature, filter, wash the filtered solid with methanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0026] 2. Preparation of erythromycin chiral silica filler:
[0027] In a reaction bottle, add 1.0g erythromycin and 5ml anhydrous pyridine, stir, add 0.1g glycidoxylated silica gel, heat to 60°C, keep warm for 16 hours, cool to room temperature, add 1.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid with water and methanol twice, respectively, and dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0028] Example 2
[0029] 1. Preparation of glycidoxylated silica gel:
[0030] In a reaction bottle, add 5g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 20ml of ethanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 80°C, keep warm for 20 hours, cool to room temperature, filter, wash the filtered solid with ethanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0031] 2. Preparation of erythromycin chiral silica filler:
[0032] In a reaction bottle, add 1.0g erythromycin and 15ml anhydrous pyridine, stir, add 1.0g glycidoxylated silica gel, heat to 100°C, keep warm for 12 hours, cool to room temperature, add 2.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid twice with water and methanol respectively, dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0033] Example 3
[0034] 1. Preparation of glycidoxylated silica gel:
[0035] In a reaction bottle, add 2.0g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 10ml of methanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 60°C, keep warm for 16 hours, cool to room temperature, filter, wash the filtered solid with methanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0036] 2. Preparation of erythromycin chiral silica filler:
[0037] In a reaction bottle, add 1.0g erythromycin and 5ml anhydrous DMSO, stir, add 1.5g glycidoxylated silica gel, heat to 80°C, keep warm for 14 hours, cool to room temperature, add 4.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid twice with water and methanol respectively, dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0038] Example 4
[0039] 1. Preparation of glycidoxylated silica gel:
[0040] In a reaction bottle, add 2.5g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 15ml of methanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 60°C, keep warm for 10 hours, cool to room temperature, filter, wash the filtered solid with methanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0041] 2. Preparation of erythromycin chiral silica filler:
[0042] In a reaction bottle, add 1.0g erythromycin and 10ml anhydrous DMSO, stir, add 0.5g glycidoxylated silica gel, heat to 120°C, keep warm for 10 hours, cool to room temperature, add 5.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid with water and methanol twice, respectively, and dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0043] Example 5
[0044] 1. Preparation of glycidoxylated silica gel:
[0045] In a reaction bottle, add 5.0g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 20ml of ethanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 70°C, keep warm for 4 hours, cool to room temperature, filter, wash the filtered solid with ethanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0046] 2. Preparation of erythromycin chiral silica filler:
[0047] In a reaction bottle, add 1.0g erythromycin and 8ml anhydrous pyridine, stir, add 2.0g glycidoxylated silica gel, heat to 90°C, keep warm for 14 hours, cool to room temperature, add 2.5g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid with water and methanol twice, respectively, and dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0048] Example 6
[0049] 1. Preparation of glycidoxylated silica gel:
[0050] In a reaction bottle, add 0.5g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 5ml of methanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 60°C, keep warm for 18 hours, cool to room temperature, filter, wash the filtered solid with methanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0051] 2. Preparation of erythromycin chiral silica filler:
[0052] In a reaction bottle, add 1.0g erythromycin, 5ml anhydrous pyridine and 10ml anhydrous DMSO, stir, add 0.2g glycidoxylated silica gel, heat to 60°C, keep warm for 16 hours, cool to room temperature, add 3.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid twice with water and methanol respectively, dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0053] Example 7
[0054] 1. Preparation of glycidoxylated silica gel:
[0055] In a reaction bottle, add 1.0g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) and 10ml of ethanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 80°C, keep warm for 18 hours, cool to room temperature, filter, wash the filtered solid with ethanol twice, and vacuum dry the filtered solid at 60°C to constant weight to obtain glycidoxylated silica gel.
[0056] 2. Preparation of erythromycin chiral silica filler:
[0057] In a reaction bottle, add 1.0 g erythromycin and 10 ml anhydrous DMSO, stir, add 0.8 g glycidoxylated silica gel, heat to 120°C, keep warm for 10 hours, cool to room temperature, add 1.0 g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid twice with water and methanol respectively, dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0058] Example 8
[0059] 1. Preparation of glycidoxylated silica gel:
[0060] In a reaction bottle, add 1.0g of γ-glycidyloxypropyltrimethoxysilane (γ-GPS), 5ml of methanol and 10ml of ethanol, stir until completely dissolved, add 1.0g of spherical porous silica gel, protect with nitrogen, heat to 70°C, keep warm for 16 hours, cool to room temperature, filter, wash the filtered solid with methanol twice, and dry the filtered solid in vacuum at 60°C to constant weight to obtain glycidoxylated silica gel.
[0061] 2. Preparation of erythromycin chiral silica filler:
[0062] In a reaction bottle, add 1.0g erythromycin, 10ml anhydrous pyridine and 5ml anhydrous DMSO, stir, add 1.5g glycidoxylated silica gel, heat to 80°C, keep warm for 14 hours, cool to room temperature, add 4.0g trimethylsilyl chloride, stir at room temperature for 6 hours, filter, wash the filtered solid twice with water and methanol respectively, dry the filtered solid in vacuum at 60°C to constant weight to obtain erythromycin chiral silica gel filler.
[0063] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.
Claims
1. An erythromycin chiral silica filler, characterized in that: It includes silica gel, γ-glycidoxypropyltrimethoxysilane (γ-GPS) and erythromycin molecules bonded to the surface of silica gel, wherein the chemical structure of γ-GPS is: The chemical structure of erythromycin is:
2. A method for preparing the erythromycin chiral silica gel filler according to claim 1, characterized in that: The following steps are involved: Step 1: preparing glycidoxylated silica gel: dissolving γ-glycidoxypropyltrimethoxysilane (γ-GPS) in a first solvent, adding silica gel, heating for reaction, cooling, filtering, washing, and drying to obtain glycidoxylated silica gel; Step 2: Prepare erythromycin chiral silica gel filler: stir the second solvent and erythromycin, then add the glycidoxylated silica gel, heat to react, cool, add trimethylsilyl chloride to perform end-capping treatment, seal the remaining hydroxyl groups on the surface of the silica gel, filter, wash the filtered solid with water and methanol respectively, dry the filtered solid to obtain erythromycin chiral silica gel filler.
3. The method for preparing the erythromycin chiral silica gel filler according to claim 2, characterized in that: In step 1, the first solvent is methanol and / or ethanol, and the silica gel is spherical porous silica gel.
4. The method for preparing the erythromycin chiral silica gel filler according to claim 3, characterized in that: In step 1, the reaction temperature is 60-80° C., the reaction pressure is normal pressure, the reaction time is 4-28 hours, and inert gas protection is required during the reaction.
5. The method for preparing the erythromycin chiral silica gel filler according to claim 4, characterized in that: In step 1, the mass ratio of silica gel to γ-glycidyloxypropyltrimethoxysilane (γ-GPS) is 1:0.5-5, and the mass volume ratio of γ-glycidyloxypropyltrimethoxysilane (γ-GPS) to the first solvent is 1:5-20.
6. The method for preparing the erythromycin chiral silica gel filler according to claim 2 or 5, characterized in that: In step 2, the second solvent is pyridine and / or DMSO.
7. The method for preparing the erythromycin chiral silica gel filler according to claim 6, characterized in that: The second solvent used in step 2 is an anhydrous solvent.
8. The method for preparing the erythromycin chiral silica gel filler according to claim 7, characterized in that: In step 2, the reaction temperature is 60-120° C. and the reaction time is 10-16 hours.
9. The method for preparing the erythromycin chiral silica gel filler according to claim 8, characterized in that: In step 2, the mass volume ratio of erythromycin to the second solvent is 1:5-15, the mass ratio of erythromycin to glycidoxylated silica gel is 1:0.1-2; and the mass ratio of erythromycin to trimethylchlorosilane is 1:1-5.