Carbohydrate ion derivative as well as preparation method and application thereof

Abstract

The invention discloses a saccharide ion derivative and a preparation method and application thereof, and belongs to the technical field of chemistry. On one hand, the invention provides a preparation method of the saccharide ion derivative, and on the other hand, the invention provides the saccharide ion derivative obtained by the method and application thereof. The saccharide ion derivative obtained by the preparation method disclosed by the invention is clear in structure, has unique structural characteristics, overcomes the defects of the prior art, and is beneficial to research and biological application; reaction conditions are mild and environment-friendly, the preparation cost is reduced, product industrialization is facilitated, the affinity, solubility, functionality and the like of saccharides are changed, the multipurpose application space of biology and materials is expanded, the saccharide ion derivative prepared by the preparation method can be used as an auxiliary or directly used, side effects such as hemolysis and kidney poison can be avoided, raw materials are renewable, and the method is suitable for industrial production. And the product is degradable and environment-friendly.

Description

Technical Field

[0001] The invention belongs to the field of chemistry, and in particular relates to a carbohydrate ion derivative and a preparation method and application thereof. Background Art

[0002] Proteins, nucleic acids and carbohydrates are the three types of macromolecules that make up life. The research on proteins and nucleic acids has become a hot topic in life sciences. Carbohydrates are the last bastion that humans need to conquer in life sciences. Sugar is the largest biomass in nature, and sugar chains are the largest biological information library in nature. It is divided into polysaccharides, oligosaccharides and monosaccharides. Polysaccharides such as cellulose and chitosan; cellulose is a chain polysaccharide of β-1,4-linked D-pyranose glucose units, the most widely distributed and most abundant polysaccharide in nature, accounting for more than 50% of the carbon content in the plant kingdom. Oligosaccharides such as cyclodextrin (CD) contain 6, 7, 8 or more α-1,4-linked D-pyranose glucose units, which are called α-CD, β-CD and γ-CD respectively. The outer surface of CD molecules is hydrophilic, and the central cavity is lipophilic, showing excellent recognition and loading capabilities for hydrophobic functional factors. CD chemical derivatives have increased water solubility, reduced toxicity, increased affinity, etc.

[0003] Charged sugars are obtained by chemical modification to produce derivatives with positively or negatively charged groups. These charged groups can interact with oppositely charged guest molecules through electrostatic interactions, thereby forming charged supramolecular assemblies and enhancing their stability, which can be applied to various biological practices, including drug delivery, gene therapy, sensing and diagnosis. In terms of materials, polysaccharide ion derivatives can be used as antibacterial agents, electrochemical materials, oil recovery aids, etc.

[0004] Sugars have many hydroxyl groups, and the direct reaction activity of hydroxyl groups is low. The constraints of the CD cone space structure make the hydroxyl reaction of cyclodextrin a challenge. There is a significant difference between the hydroxyl reactions of conical oligosaccharides and chain oligosaccharides. After condition exploration and optimization, the present invention obtains an environmentally friendly and mild reaction condition suitable for chain and ring sugars, which is conducive to reducing preparation costs and facilitating industrialization.

[0005] The existing ion derivatization reaction technology needs to be improved, such as the use of ionic liquids, DMSO organic solvents that are difficult to handle, etc., which are high in cost, troublesome post-processing, and low in conversion rate. Summary of the invention

[0006] In view of the problems existing in the prior art, the present invention uses the one-pot method for ionization of sugars under aqueous conditions for the first time, using inorganic bases as reactants, solvents, and catalysts; the reaction conditions are mild, the degree of substitution is moderate, and the conversion rate is high. Ionic derivatives such as cellulose, cyclodextrin, and trehalose are obtained. The product structure is characterized; and the antibacterial experiment of polysaccharide derivatives and the application evaluation of pharmaceutical adjuvants are carried out, which is specifically achieved through the following technical solutions:

[0007] The present invention firstly provides a method for preparing carbohydrate ion derivatives. The method adopts a one-pot carbohydrate ionization method under aqueous conditions, specifically, carbohydrate substances and ion derivatives are reacted in an inorganic alkaline solution to prepare carbohydrate ion derivatives.

[0008] Furthermore, the carbohydrate material includes polysaccharides, oligosaccharides and monosaccharides. The polysaccharide is cellulose, the oligosaccharide is cyclodextrin or trehalose, and the monosaccharide is alkyl galactose.

[0009] Furthermore, the group of the ionic derivative is any one of sodium alkyl sulfonate and alkyl quaternary ammonium salt.

[0010] Furthermore, the inorganic alkali solution is 4-10% sodium hydroxide solution, and the sodium hydroxide is used as a catalyst, a solvent and a raw material participating in the reaction.

[0011] Furthermore, the carbohydrate ion derivatives are based on monosaccharide units, and the groups are randomly substituted with a degree of substitution of 1-3.

[0012] The second aspect of the present invention provides a carbohydrate ion derivative obtained by the above preparation method.

[0013] Furthermore, the cyclodextrin anion derivative with a degree of substitution of 1 among the above-mentioned carbohydrate ion derivatives is used as a pharmaceutical adjuvant.

[0014] Furthermore, the cellulose cationic derivative with a degree of substitution of 2 among the above-mentioned carbohydrate ion derivatives is used as an antibacterial agent.

[0015] Furthermore, the application is specifically the antibacterial and moisturizing effects in leather artifacts.

[0016] Furthermore, the alkyl galactose cationic derivatives with a substitution degree of 2 among the above-mentioned carbohydrate ion derivatives are used as antibacterial agents for daily necessities.

[0017] Ionic derivatives of carbohydrates can improve the solubility of polysaccharides or cyclic oligosaccharides, change the polarity of carbohydrates, reduce their biological toxicity, change their aggregation state, etc. Cyclodextrins include α-CD, β-CD and γ-CD. The groups of ionic derivatives include sodium alkyl sulfonate, alkyl quaternary ammonium salt, and the groups are randomly substituted, with the highest degree of substitution of monosaccharide units being 3.

[0018] The carbohydrate ion derivative obtained by the preparation method of the present invention has a clear structure and unique structural characteristics, which fills the shortcomings of previous technologies and is beneficial to research and biological applications; the reaction conditions are mild and environmentally friendly, which reduces the preparation cost and is beneficial to product industrialization, while changing the affinity, solubility and functionality of carbohydrates, etc., expanding the multi-purpose application space of biology and materials. The carbohydrate ion derivative obtained by the preparation method of the present invention can be used as an auxiliary agent or directly used, which can avoid side effects such as hemolysis and kidney toxicity, and the raw materials are renewable, the products are degradable, and are environmentally friendly, which is particularly suitable for the concepts of cultural relics protection, environmental protection and sustainable development. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the standard curve of shikonin. DETAILED DESCRIPTION

[0020] The present invention is further described below in conjunction with specific embodiments to facilitate a better understanding of the present technical solution.

[0021] Example 1

[0022] Weigh 2g of crude cellulose after extracting the water-soluble matter, measure 8%wt NaOH solution, add 2,3-epoxypropyltrimethylammonium chloride (B), control the reaction at 70°C for 24 hours, then cool and neutralize with dilute hydrochloric acid solution, dialyze with a 2000 molecular weight dialysis bag for about 72 hours, freeze-dry, and weigh. The yield is 71%. After elemental analysis, the degree of substitution is 2. Product 1 is obtained.

[0023] Experiments have shown that the NaOH / B molar ratio of the reaction system is 1.1 / 1, and the B / monosaccharide molar ratio is 5 / 1.

[0024] For the Fourier infrared spectrum of polysaccharides: After the sample is freeze-dried, the raw material is used for comparison, and the infrared test is performed using the KBr tablet method. The test is performed on an infrared spectrometer, and the test is scanned continuously 32 times, and the instrument resolution is 4cm -1 , analysis range 400~4000cm -1 , wave number accuracy is 0.01cm -1 The product showed some specific absorption peaks compared with the raw material. 3420m -1 The strong absorption peak nearby is caused by the stretching vibration of the OH molecule; 2961cm -1 The absorption peak at 1639cm is caused by the methylene stretching vibration; -1 The peaks near reflect the hydrogen bond absorption peaks formed by hydroxyl groups and free water; these three absorption peaks constitute the characteristic absorption peaks. -1 At, generated by CO stretching vibration; wave number 1482cm -1 and 1360cm -1There is a characteristic absorption peak nearby, corresponding to the methyl absorption peak of CH3-N, proving that the cationic group has been successfully introduced into the macromolecule.

[0025] Example 2

[0026] Weigh 2g of β-cyclodextrin, measure 10%wt NaOH solution, add sodium 3-chloro-2-hydroxypropanesulfonate (A), control the reaction at 70°C for 12 hours, then cool and neutralize with dilute hydrochloric acid solution, dialyze with a 500 molecular weight dialysis bag for about 72 hours, freeze-dry, and weigh. The yield is 83%. Sulfur element analysis shows that the degree of substitution is 1. Experiments show that the NaOH / A molar ratio of the reaction system is 1 / 1, and the A / cellulose molar ratio is 5 / 1, and the reaction degree of substitution of the product is controlled to be 1. Obtain product 2.

[0027] For monosaccharides and oligosaccharides, NMR is used to characterize their structures. NMR of cyclodextrin derivative product 2: 1 H NMR (400MHz, D2O) δ4.95 (d, J = 3.7Hz), 4.15 (dd, J = 4.0, 1.9Hz), 3.89-3.71 (m), 3.56-3.42 (m), 3.17-2.85 (m). 13 CNMR(101MHz,D2O)δ101.95,81.21,73.17,71.96,68.16,67.28,64.70,60.34,54.26,48.48.

[0029] Example 3

[0030] Weigh 2g of 1-octylgalactose, measure 10%wt NaOH solution, add sodium 3-chloro-2-hydroxypropanesulfonate (A), control the reaction at 70°C for 12 hours, then cool and neutralize with dilute hydrochloric acid solution, dialyze with a 200 molecular weight dialysis bag for about 72 hours, freeze-dry, and weigh. The yield is 79%. The NaOH / A molar ratio of the reaction system is 1 / 1, and the A / cellulose molar ratio is 5 / 1. After elemental analysis, it is determined that the degree of substitution is 1. The NMR of the obtained product 3 is: 1 H NMR(400MHz,D2O)δ6.61(d,J=12.2Hz),6.47(s),4.26(s),4.14(s),4.01-3.85(m),3.62(d,J= 30.3Hz),3.38-3.34(m),3.05(dd,J=33.5,13.2Hz),1.88(s,1H),1.59(s),1.25(s),0.84(s). 13C NMR (101MHz, D2O) δ98.14,73.57-68.45,66.72-59.60,56.13-51.92,31.38,28.45,25.07,21.6,13.93.

[0031] Example 4

[0032] 1) Preparation of product 1 film: Dissolve in water according to the following ratio (weight ratio, the ratio is determined by screening), product 1: chitosan: gelatin: water = 9:0.2:0.2:50, stir for 2 hours, vacuum degas the mixture, pour it on a plastic sheet to form a film, and form a film after drying. The film is transparent, has certain flexibility and tensile strength. The tensile strength of the film is 61.05±0.4MPa, and the elongation at break is 72.01±1.3%. The hygroscopicity in an environment with a relative humidity of 43% and the water retention in dry silica gel are higher than those of the film made with the original cellulose.

[0033] 2) Antibacterial experiment of product 1 film: adjust the density of bacterial suspension (select Aspergillus niger, Aureobasidium budding and fungi on moldy leather) and evenly apply it on the condensed plate culture medium. First sterilize the film (6mm in diameter) and stick it on the surface of the plate. One piece of sterilized pure cellulose filter paper is used as a blank control. Parallel 3 experiments are carried out. Culture on the plate culture medium at 25℃ for 48h, and the size of the inhibition zone is determined. The antibacterial effect is shown in Table 1.

[0034] Table 1: Antibacterial effect of thin composite films containing product 1

[0035]

[0036] Note: The standard for the inhibition zone test is: if the inhibition zone is larger than 20 mm, it is extremely sensitive; if it is 15 to 20 mm, it is highly sensitive; if it is 10 to 15 mm, it is moderately sensitive; if it is 7 to 10 mm, it is low sensitive; if it is smaller than 7 mm, it is insensitive.

[0037] As shown in Table 1, the thin composite film containing product 1 showed a moderately sensitive antibacterial effect on the three types of fungi. The antibacterial effect on Aspergillus niger was slightly stronger. The thin composite film containing product 1 is composed of carbohydrates and their derivatives, and proteins. It is more ecologically compatible with the protected leather cultural relics, can be degraded, and has a certain water retention capacity, preventing the cultural relics from drying and cracking, etc. It will not cause secondary pollution to the cultural relics, and is in line with the natural concept of traditional culture.

[0038] 3) Antibacterial experiment of product 3: adjust the density of bacterial suspension (select Aspergillus niger, Aureobasidium budding and fungi on moldy orange peel) and evenly apply it on the condensed plate culture medium. Prepare 8% product 3 aqueous solution (optimal concentration determined by screening) to soak filter paper (diameter 6mm), dry, sterilize, and stick it on the surface of the plate, one of which is sterilized pure cellulose filter paper for blank control, parallel 3 experiments, culture on the plate culture medium at 25°C for 48h, and determine the size of the inhibition zone. The antibacterial effect is shown in Table 2.

[0039] Table 2: Antibacterial effect of 8% product 3

[0040]

[0041] It can be seen from Table 2 that the 8% product 3 showed moderately sensitive antibacterial effects on the three types of fungi, and had a slightly stronger antibacterial effect on Aspergillus niger.

[0042] Example 5

[0043] The water solubility of cyclodextrin derivative sample 2 is 53.2 mg / mL (β-cyclodextrin solubility: 18.5 mg / mL). The water solubility of cyclodextrin is significantly improved after the reaction.

[0044] 1) Inclusion reaction: Dissolve sample 2 in 5 mL of deionized water to obtain solution 1. Dissolve 7 mg of shikonin in 1.0 mL of ethanol. The molar ratio of sample 2 to shikonin is 2:1. Add the shikonin solution dropwise to solution 1. React at 60°C and 500 rpm for 2 hours. After the reaction, cool to room temperature, centrifuge at 3000 rpm for 10 minutes, obtain the supernatant, freeze-dry for 24 hours, and weigh the dried solid.

[0045] 2) Encapsulation efficiency: 1.0 mL of the obtained inclusion complex solution was dissolved in anhydrous ethanol and added to a 10 mL volumetric flask, and then ultrasonically treated for 30 minutes to remove the cyclodextrin derivatives to obtain the total concentration of shikonin solution Ct.

[0046] 1.0 mL of the obtained inclusion complex solution was added to a 10 mL volumetric flask filled with deionized water, and then the two samples were centrifuged at 4000 rpm for 10 minutes to obtain a free shikonin solution Cf. The absorbance of the supernatant was measured at 516 nm, and the standard curve of shikonin (the equation Abs = 0.0187 [C] -0.0115 was obtained, where Abs is the absorbance, [C] is the drug concentration (R 2 =0.9939). The standard curve of shikonin is shown in Figure 1 .

[0047] The drug encapsulation efficiency (EE%) was calculated by the following formula: EE (%) = |Ct-Cf| / Ct×100

[0048] (where Ct is the total concentration of shikonin and Cf is the concentration of free shikonin).

[0049] Through the above test, the EE% of the encapsulation rate of sample 2 (DS=1) and shikonin was 49%, indicating that cyclodextrin derivatives can encapsulate fat-soluble drugs with a relatively high encapsulation rate; at the same time, since the solubility of cyclodextrin derivatives is greatly improved, the solubility of fat-soluble drugs is also improved.

[0050] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A method for preparing a carbohydrate ion derivative, characterized in that: The preparation method adopts a one-pot method to ionize sugars under aqueous conditions, specifically, sugar substances and ion derivatives are reacted in an inorganic alkaline solution to obtain sugar ion derivatives.

2. The method for preparing the carbohydrate ion derivative according to claim 1, characterized in that: The sugar substances include polysaccharides, oligosaccharides and monosaccharides. The polysaccharides are cellulose, the oligosaccharides are cyclodextrin or trehalose, and the monosaccharides are alkyl galactose.

3. The method for preparing a carbohydrate ion derivative according to claim 1, characterized in that: The group of the ionic derivative is any one of sodium alkyl sulfonate and alkyl quaternary ammonium salt.

4. The method for preparing a carbohydrate ion derivative according to claim 1, characterized in that: The inorganic alkali solution adopts 4-10% sodium hydroxide solution, and the sodium hydroxide is used as a catalyst, a solvent and a raw material participating in the reaction.

5. The method for preparing a carbohydrate ion derivative according to claim 1, characterized in that: The carbohydrate ion derivatives are based on monosaccharide units, with groups randomly substituted and a degree of substitution of 1-3.

6. A carbohydrate ion derivative obtained by the preparation method according to any one of claims 1 to 5.

7. The carbohydrate ion derivative according to claim 6, characterized in that Application of cyclodextrin anion derivatives with a degree of substitution of 1 in pharmaceutical adjuvants.

8. The carbohydrate ion derivative according to claim 6, characterized in that Application of cellulose cationic derivatives with a degree of substitution of 2 as antimicrobial agents.

9. The carbohydrate ion derivative according to claim 8, characterized in that The application is specifically the antibacterial and moisturizing effect in leather artifacts.

10. The carbohydrate ion derivative according to claim 6, characterized in that Application of alkylgalactose cationic derivatives with a degree of substitution of 2 as antimicrobial agents for daily necessities.

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