CN116554363A - Torone-chitosan derivative and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the technical field of antibacterial substances, and discloses a tropolone-chitosan derivative and a preparation method and application thereof. The structural formula of this tropolone-chitosan derivative is as shown in formula (1): Wherein, R ₁ represents at least one of -H, hydroxyl-containing alkyl or carboxyl-containing alkyl; R ₂ represents -H, hydroxyl-containing alkyl, carboxyl-containing alkyl or carbonyl-containing alkyl At least one; x and y independently represent positive integers; substances containing tropolone structural units are replaced by mannitol. The tropolone-chitosan derivative has good antibacterial properties, has good antibacterial effects on Gram-positive bacteria, Gram-negative bacteria, yeast and mold, and has good high temperature stability and solubility Good, very conducive to the application of tropolone-chitosan derivatives in the fields of medicine, cosmetics or food.

Description

A kind of tropolone-chitosan derivative and its preparation method and application

This application is a divisional application with an application date of May 21, 2021, an application number of 202110555368.4, and an invention titled "a tropolone-chitosan derivative and its preparation method and application".

technical field

The invention belongs to the technical field of antibacterial substances, and in particular relates to a tropolone-chitosan derivative and a preparation method and application thereof.

Background technique

Chitin is a natural polymer compound, which is formed by connecting N-acetyl-D-glucosamine and D-glucosamine through β-(1-4) glycosidic bonds. It can be partially or completely deacetylated to obtain the commonly used chitosan. Most chitosan on the market is obtained from shrimp and crab shells through a series of treatments (acid washing or alkali washing). It is non-toxic and tasteless, and has been widely used in food, medicine, cosmetics and other industries with excellent moisturizing properties Sex, immunomodulatory activity, antibacterial and other advantages. However, chitosan also has some limitations. First, chitosan is insoluble in water and can only be dissolved under acidic conditions; second, as an antibacterial agent, it needs to be added to a higher concentration, resulting in low cost performance; The positive charge carried is easy to flocculate with the anion raw materials in the system, resulting in poor stability of the system.

Chitosan contains amino groups and hydroxyl groups, and can be chemically modified under relatively mild reaction conditions. In the fields of medicine and cosmetics, water-soluble chitosan derivatives (such as carboxymethyl chitosan) are commonly used. However, in the existing related technologies, the antibacterial properties of the modified chitosan derivatives are often poor, resulting in a better antibacterial effect, usually need to add a higher concentration of chitosan derivatives, which is not conducive to Applications of chitosan derivatives. In addition, the existing chitosan derivatives have poor stability to high temperature (for example, 120° C.), and the antibacterial performance of the chitosan derivatives treated at high temperature decreases significantly.

Therefore, need to provide a kind of new chitosan derivative urgently, the antibacterial property of this chitosan derivative is significantly improved than before, and stability is good, and application cost is lower, and this contributes very much to the development of chitosan derivative. application.

Contents of the invention

The present invention aims to solve at least one of the technical problems in the above-mentioned prior art. For this reason, the present invention proposes a kind of tropolone-chitosan derivative and its preparation method and application, described tropolone-chitosan derivative has good antibacterial property, especially through high temperature (for example greater than 140 ℃ , specifically have 150 ℃, 180 ℃) processed tropolone-chitosan derivatives, still have good antibacterial properties, as seen the tropolone-chitosan derivatives of the present invention have good stability to temperature .

The inventive concept of the present invention: the present invention uses chitosan or chitosan derivatives, and tropolone or substances containing tropolone structural units as reaction raw materials, and these two types of reactants react to obtain a Compound (also can be referred to as a kind of salt), the compound described in the present invention has good antibacterial property, especially has good high temperature resistance stability (such as 150 ℃, 180 ℃), good solubility, low preparation cost, very Facilitate the application of the compound.

The first aspect of the present invention provides a tropolone-chitosan derivative.

Concrete, a kind of tropolone-chitosan derivative, the structural formula of described tropolone-chitosan derivative is as shown in formula (1):

Wherein, _R represents at least one of -H, hydroxyl-containing alkyl or carboxyl-containing alkyl;

_R represents at least one of -H, hydroxyl-containing alkyl, carboxyl-containing alkyl or carbonyl-containing alkyl;

R ₃ , R ₄ , R ₅ , R ₆ , and R ₇ each independently represent at least one of -H, hydroxyl, carboxyl, alkenyl, substituted alkyl or unsubstituted alkyl;

The x and y independently represent positive integers.

Preferably, the substituted alkyl groups independently represented by R ₃ , R ₄ , R ₅ , R ₆ , and R ₇ include alkylcarbonyl, alkoxy, alkyl ester, and alkylhydroxy, such as -OCH ₃ , -COCH ₃ .

Preferably, the R ₁ is at least one selected from -H, -CH ₂ COOH, -CH ₂ CH ₂ OH or -CH ₂ CH ₂ (OH)CH ₃ .

Preferably, the R ₂ is at least one selected from -H, -COCH ₃ , CH ₂ COOH or CH ₂ CH ₂ (OH)CH ₃ .

Preferably, the R ₃ is at least one selected from -H, -OH or -CH(CH ₃ ) ₂ .

Preferably, the R ₄ is at least one selected from -H, -CH(CH ₃ ) ₂ , -CH═CH ₂ CH ₃ , -COCH ₃ or -COOH.

Preferably, the R ₅ is at least one selected from -H, -CH(CH ₃ ) ₂ , -CH ₂ CH=C(CH3) ₂ or -CH ₂ CH ₂ C(OH)(CH ₃ ) ₂ .

Preferably, the R ₆ is at least one selected from -H or -OH.

Preferably, the R ₇ is at least one selected from -H or -OCH ₃ .

Preferably, the value range of x is 1-50000; further preferably, the value range of x is 1-46300.

Preferably, the value range of y is 1-50000; further preferably, the value range of y is 1-46300.

Preferably, the raw material components for preparing the tropolone-chitosan derivatives include chitosan or chitosan derivatives, and tropolone or substances containing tropolone structural units.

Preferably, the deacetylation degree of the chitosan derivative is between 60%-100%, preferably between 80%-99%, and the molecular weight of the chitosan derivative can be between 100-10000000Da.

Preferably, the lower the amino substitution degree of the chitosan derivative, the better, preferably less than 10%. Because the lower the degree of amino substitution, more amino groups can be released, and the chitosan derivative can react with tropolone or a substance containing tropolone structural unit, thereby enhancing the antibacterial property of the final product.

Preferably, the degree of substitution of hydroxyl groups at the 3rd and 6th positions of the chitosan derivative can be 1-100%, preferably 60-100%. This helps to increase the water solubility of chitosan derivatives.

Preferably, the chitosan derivative is selected from at least one of carboxymethyl chitosan, carboxyethyl chitosan, carboxyethyl chitosan, potassium salt of chitosan or sodium salt of chitosan kind.

Preferably, the substance containing tropolone structural unit is selected from the group consisting of α-thujen, β-thujen, γ-thujen, thujol, 4-acetyl-tropolone, preconditions, 3,6-Dihydroxy-5-oxo-1,3,6-cycloheptatriene-1-carboxylic acid, β-axocenol, α-thukiol, iso-porphyrin or mannitol (or D - at least one of mannitol).

The second aspect of the present invention provides a preparation method of tropolone-chitosan derivatives.

Concrete, a kind of preparation method of tropolone-chitosan derivative, comprises the following steps:

Dissolving chitosan or chitosan derivatives in a solvent, then adding tropolone or a substance containing tropolone structural units, heating and reacting to prepare the tropolone-chitosan derivatives.

Preferably, the mass ratio of the chitosan or chitosan derivative to tropolone or the substance containing tropolone structural unit is 5:(0.8-5); further preferably, the chitosan or chitosan The mass ratio of the polysaccharide derivative to tropolone or a substance containing a tropolone structural unit is 5:(1-5). The less the amount of tropolone or the substance containing the tropolone structural unit is, the lower the preparation cost is.

Preferably, the solvent contains water of organic acid, which helps the dissolving of chitosan.

Preferably, the temperature of the heating reaction is 55-90°C; further preferably, the temperature of the heating reaction is 58-65°C.

Preferably, the time for the heating reaction is more than 1 hour; more preferably, the time for the heating reaction is 1-3 hours.

Preferably, after the heating reaction is finished, the product is freeze-dried to obtain a powdery product.

Preferably, after the heating reaction is finished, the product is formulated into a solution with a mass fraction of 2-35%; further preferably, the product is formulated into a solution with a mass fraction of 3-30%.

Preferably, after the heating reaction is finished, the tropolone-chitosan derivative can be purified by washing with alcohol (such as ethanol). Tropolone-chitosan derivatives are insoluble in ethanol. Chitosan or chitosan derivatives, and tropolone or substances containing tropolone structural units are easily soluble in ethanol.

The third aspect of the present invention provides the application of a tropolone-chitosan derivative.

Application of the above-mentioned tropolone-chitosan derivatives in the preparation of antibacterial substances.

The application of the tropolone-chitosan derivative in the fields of medicine, cosmetics or food.

Compared with the prior art, the beneficial effects of the present invention are as follows:

(1) The present invention takes chitosan or chitosan derivatives, and tropolone or the material containing tropolone structural unit as reaction raw materials, and these two types of reactants react to obtain tropolone-shell by electrostatic interaction. The polysaccharide derivatives, the tropolone-chitosan derivatives in the present invention have good antibacterial properties, and have good antibacterial effects on Gram-positive bacteria, Gram-negative bacteria, yeast and mold.

(2) The tropolone-chitosan derivatives described in the present invention have good high temperature stability (such as 150°C, 180°C), and good solubility, which is very beneficial to the application of the tropolone-chitosan derivatives .

(3) The preparation process and purification process of the tropolone-chitosan derivative described in the present invention are simple, the preparation cost is low, and can be widely used in the fields of medicine, cosmetics or food.

Description of drawings

Fig. 1 is the proton nuclear magnetic resonance spectrogram of the tropolone-chitosan derivative that embodiment 1 makes.

Detailed ways

In order to make those skilled in the art understand the technical solution of the present invention more clearly, the following examples are listed for illustration. It should be pointed out that the following examples do not limit the protection scope of the present invention.

Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial channels, or can be obtained by existing known methods.

Embodiment 1: the preparation of tropolone-chitosan derivative

The structural formula of the tropolone-chitosan derivative that present embodiment 1 makes is as shown in formula (2):

Wherein R ₁ represents -H, R ₂ represents carboxymethyl, the value of x is 970, and the value of y is 330.

The preparation method of above-mentioned tropolone-chitosan derivative, comprises the following steps:

Take by weighing 10g sodium carboxymethyl chitosan (the molecular weight of sodium carboxymethyl chitosan is 280000Da, the degree of deacetylation is 85%, the degree of O substitution is 75%, and the degree of N substitution is 5%), dissolved in 1L of water, And add 2g of β-thujen, stir magnetically at 150 rpm, and heat the reaction at 60°C for 1 hour, then freeze-dry, and grind the product into powder, wash with ethanol for 5 times, then freeze-dry, and grind into powder again , to prepare tropolone-chitosan derivatives.

Fig. 1 is the proton nuclear magnetic resonance spectrogram of the tropolone-chitosan derivative that embodiment 1 makes. What " 1 " in Fig. 1 represents is the hydrogen nuclear magnetic resonance spectrum of beta-thujenin, and what "2" represents is the hydrogen nuclear magnetic resonance spectrum of the tropolone-chitosan derivative that embodiment 1 makes. The upper left corner in Figure 1 is an enlargement of chemical shifts of 6.8-7.5 ppm.

¹ H-NMR analysis of the tropolone-chitosan derivative prepared in Example 1 confirmed the structure of the tropolone-chitosan derivative product prepared in Example 1. The signals in the ¹ H-NMR spectrum represent protons in different electronic environments and thus can be used to identify specific chemical groups, as well as changes in electron density.

The H NMR spectrum of β-thujenin ("1" in Fig. 1) shows signals of different protons associated with the carbon atoms of this molecule. The proton distribution is as follows (ppm): 1.17-1.18 (CH ₃ , isopropyl group of β-thujenin), 2.9 (CH, isopropyl group of β-thujenin), 7.19-7.5 (CH, aromatic ring carbon 3 , 4, 5, 7).

The peak at 4.7 ppm is the solvent peak. The H NMR spectrum of the tropolone-chitosan derivative ("2" in Fig. 1) prepared in Example 1 shows the proton signal combined with carboxymethyl chitosan and β-orientinin. The protons (ppm) bound to carboxymethyl chitosan are: 1.98 (CH, non-deacetylated), 2.68 (CH, glucosamine ring carbon 2), 3.31-3.86 (CH, glucosamine ring carbon 3, 4 and 6), 3.91 (CH, carboxymethylated). The number of protons combined with β-thujen is arranged in the following order (ppm): 1.14-1.1 (CH ₃ , isopropyl group of β-thujen), 2.78 (CH, isopropyl group of β-thujen) .

Figure 1 confirms that both β-orientinin and carboxymethyl chitosan structures exist in tropolone-chitosan derivatives, and no covalent modification occurs. Figure 1 also confirms that there is an interaction between β-thujen and carboxymethyl chitosan, because the protons of the aromatic ring of β-thujen (7.19-7.5ppm in its natural form) have moved to the front field (6.80-7.24ppm in the tropolone-chitosan derivative prepared in Example 1). This shift can be seen in the upper left corner of Figure 1 in the zoomed-in section for chemical shifts of 6.8-7.5 ppm.

Changes in the H NMR spectrum are related to changes in electron density. Figure 1 shows that the electron density of the aromatic ring of β-orientinin is increased. The β-orientinin structure forms an anion that delocalizes and stabilizes the negative charge through its resonance structure.

Embodiment 2: the preparation of tropolone-chitosan derivative

A preparation method of tropolone-chitosan derivatives, comprising the following steps:

Weigh 10g carboxymethyl chitosan (the molecular weight of carboxymethyl chitosan is 300000Da, the degree of deacetylation is 85%, the degree of O substitution is 75%, the degree of N substitution is 5%), dissolve it in 1L of water, and add 1.5g gamma-thujenin, magnetically stirred at 150 rpm, and heated at 70°C for 1.5 hours, then freeze-dried, and the product was ground into powder, washed 5 times with ethanol, then freeze-dried, and ground into powder again, Tropolone-chitosan derivatives were prepared.

Embodiment 3: the preparation of tropolone-chitosan derivative

A preparation method of tropolone-chitosan derivatives, comprising the following steps:

Take by weighing 100g carboxybutyl chitosan (the molecular weight of carboxybutyl chitosan is 300000Da, the degree of deacetylation is 90%, the degree of O substitution is 75%, the degree of N substitution is 6%), dissolve in 1L of water, and add 40g D-mannitol, in 150 rev/min magnetic stirring, and 1.2 hours at 75 ℃ of heating reactions, prepared tropolone-chitosan derivatives, prepared tropolone-chitosan derivatives were reacted The mass concentration in the final solution was 14%.

Embodiment 4: the preparation of tropolone-chitosan derivative

A preparation method of tropolone-chitosan derivatives, comprising the following steps:

Weigh 10g carboxyethyl chitosan (the molecular weight of carboxyethyl chitosan is 350000Da, the degree of deacetylation is 80%, the degree of O substitution is 75%, the degree of N substitution is 5%), dissolve it in 1L of water, and add 5g of β-axacine, stirred magnetically at 150 rpm, and heated at 80°C for 1 hour, then freeze-dried, and the product was ground into powder, washed with ethanol for 5 times, then freeze-dried, and ground into powder again to prepare Tropolone-Chitosan Derivatives.

Embodiment 5: the preparation of tropolone-chitosan derivative

A preparation method of tropolone-chitosan derivatives, comprising the following steps:

Weigh 10g of chitosan (the molecular weight of chitosan is 250,000Da, the degree of deacetylation is 92%, the degree of O substitution is 75%, and the degree of N substitution is 5%), dissolve it in 1L of water, add 10mL of acetic acid, and then add 8g of α-thugophenol, magnetically stirred at 150 rpm, and heated at 60°C for 1.5 hours, then freeze-dried, and the product was ground into powder, washed with ethanol for 5 times, then freeze-dried, and ground into powder again to prepare Tropolone-Chitosan Derivatives.

Embodiment 6: the preparation of tropolone-chitosan derivative

Compared with Example 5, in Example 6, tropolone was used to replace α-Thukiol, and the rest of the preparation process was the same as in Example 5.

Product Effect Test

1. Antibacterial effect test

Get the tropolone-chitosan derivative that embodiment 1-6 makes, and carboxymethyl chitosan, β-thujen (carboxymethyl chitosan, β-thujen as contrast), test It is effective against bacteria (including Gram-positive bacteria and Gram-negative bacteria, wherein Gram-positive bacteria include Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Lactobacillus plantarum, and Gram-negative bacteria include large intestine Bacillus, Pseudomonas aeruginosa), yeast (Candida albicans), mold (A. , 300ppm).

The cultured system is a commercially available nutrient broth (provided by Guangdong Huankai Microbial Technology Co., Ltd., model 022010), the pH of the cultured system is 6, and the culture conditions of the bacteria are as follows: at a temperature of 36 ° C for 7 days, The culture conditions of the yeast and the mold were cultured at 28° C. for 7 days, and the results are shown in Table 1.

Table 1: antibacterial effect (data in table 1 represent MIC, unit ppm)

Remarks: "/" in Table 1 means no antibacterial effect.

It can be seen from Table 1 that the tropolone-chitosan derivatives prepared in Examples 1-6 of the present invention have better antibacterial effects than carboxymethyl chitosan. From Table 1, it can be seen that β-abrevitin also has good antibacterial properties, but compared with the antibacterial properties of the tropolone-chitosan derivatives obtained in Examples 1-6 of the present invention, β-orientinin The antibacterial performance of the antibacterial property is still relatively weak (except Pseudomonas aeruginosa), and the commercially available price of β-thujen is more than 3 times of the price of the tropolone-chitosan derivative that the embodiment of the present invention 1-6 makes , so the tropolone-chitosan derivatives prepared in Examples 1-6 of the present invention are cost-effective.

2. High temperature stability test

Get the tropolone-chitosan derivative that embodiment 1 makes, test it and process through different temperature conditions (different temperature conditions are specifically divided into: normal temperature 25 ℃ process 1 hour, high-pressure wet heat treatment (101KPa, 121 ℃, 15 minutes ), the antibacterial effect after 150° C. of normal pressure oil bath for 1 hour, and 180° C. of normal pressure oil bath for 1 hour), and then the stability of the tropolone-chitosan derivatives obtained in Example 1 can be known. The antibacterial results are shown in Table 2.

The cultured system is a commercially available nutrient broth (provided by Guangdong Huankai Microbial Technology Co., Ltd., model 022010), the pH of the cultured system is 6, and the culture conditions of the bacteria are as follows: at a temperature of 36 ° C for 7 days, The culture conditions of the yeast and mold were cultured at 28° C. for 7 days.

Table 2: antibacterial effect (data in table 2 represent minimum inhibitory concentration MIC, unit is ppm)

It can be seen from Table 2 that after treatment under conditions such as high-pressure wet heat treatment for 1 hour, 150°C treatment in an atmospheric pressure oil bath for 1 hour, and 180°C treatment in an atmospheric pressure oil bath for 1 hour, the tropolone-chitopolymer prepared in Example 1 The antibacterial effect of sugar derivatives on Gram-positive bacteria, Gram-negative bacteria, yeast and yeast is consistent with normal temperature at 25°C, showing that the tropolone-chitosan derivatives prepared in Example 1 of the present invention have good High temperature stability.

The tropolone-chitosan derivatives prepared in the remaining examples have similar high-temperature stability as the tropolone-chitosan derivatives prepared in the above-mentioned Example 1.

3. Dissolution performance test

Take the tropolone-chitosan derivatives prepared in Examples 1-6, as well as chitosan and β-orientinin, and test their solubility in water at a temperature of 20° C. The results are shown in Table 3.

Table 3: Solubility Test Results

As can be seen from Table 3, the tropolone-chitosan derivative that the embodiment of the present invention 1-6 makes has better water solubility relative to chitosan and β-thujen. The application and effect of chitosan derivatives have a very important influence.

4. Antiseptic effect test of tropolone-chitosan derivatives in cosmetics

Test strains: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa; fungi: Aspergillus niger, Candida albicans.

Medium: TSB solid medium (tryptone soy medium); SDB solid medium (Sabouraud dextrose medium); lecithin Tween 80 nutrient agar medium, tiger red medium.

The specific steps of the anti-corrosion challenge experiment are as follows:

(1) Inoculation of bacterial strains: each bacterial strain is activated and subcultured for 3 generations to make a certain concentration of bacterial suspension. The amount of bacteria contained in the sample is 1×10 ⁶ cfu/mL or 1×10 ⁶ cfu/g, and the amount of fungi contained is 1×10 ⁴ cfu/mL or 1×10 ⁴ cfu/g;

(2) Preparation of samples for inspection: Weigh 10 g of samples for inspection, add them to a conical flask equipped with glass beads and 90 mL of sterilized normal saline, fully oscillate and mix, let stand for 15 minutes, and take the supernatant as 1: 10 of the test solution, and then make a 10-fold dilution series concentration gradient test solution for use. Hydrophobic samples: Weigh 10g of the sample for testing, put it in a sterilized mortar, add 10mL of sterilized liquid paraffin, and grind it into a viscous Thick, then add 10mL sterilized Tween 80, grind until dissolved, add 70mL sterilized normal saline, mix thoroughly in a 45°C water bath to make a 1:10 suspension, and then make a 10-fold dilution series concentration The gradient test solution is ready for use;

(3) Detection of samples to be tested: Sampling and testing were carried out according to the above sampling method on 0d, 4h, 1d, 7d, 14d, 21d, and 28d ("d" means "day") after adding the bacterial solution, and the above-mentioned preparation Add 1mL of the 10-fold dilution series test solution to a sterilized plate, then pour an appropriate amount of lecithin Tween 80 nutrient agar medium or tiger red medium cooled to 45°C, shake counterclockwise to mix evenly, and do 2 for each dilution gradient. Invert the plates in parallel, and after the plates are solidified, the bacteria are counted after being incubated in an incubator at 37°C for 48 hours, and the fungi are counted after being incubated in an incubator at 28°C for 72 hours;

(4) Efficacy evaluation criteria of the anti-corrosion system:

(a) On the 28th day, if the sample contained bacteria or mold >10 ³ cfu/g (or cfu/mL), the sample could not pass the challenge test of microbial attack, indicating that the preservative system of the sample could not effectively inhibit microorganisms The product is easily contaminated by microorganisms during production, storage and use;

(b) On the 28th day, the sample contains bacteria at 10 ² cfu/g-10 ³ cfu/g (or cfu/mL), and the sample can pass the challenge test conditionally, that is, when the product contains protein or other animal and plant materials The composition is not particularly high, and the hygienic environment of the production meets the requirements, and the packaging is not prone to secondary pollution, the anti-corrosion system can be used, otherwise it cannot;

(c) On the 28th day, the sample contains bacteria at 10cfu/g-100cfu/g (or cfu/mL), indicating that the antiseptic system of the sample has a strong inhibitory effect on microorganisms. Through the challenge test, the product is in production, Not susceptible to microbial contamination during storage and use;

(d) From the 7th day, the bacteria in the sample are <10cfu/g (or cfu/mL), indicating that the antiseptic system of the sample has a strong inhibitory effect on microorganisms. It is not easy to be contaminated by microorganisms during use.

(5) Preparation of samples for inspection:

The ingredients of the test sample cream are shown in Table 4.

Table 4: Ingredients of cream for testing samples

The composition of the sample essence for inspection is shown in Table 5.

Table 5: Component list of the sample essence for inspection

The antiseptic challenge results of the tropolone-chitosan derivatives prepared in Example 1 applied to creams are shown in Table 6.

Table 6: Antiseptic challenge results of creams containing tropolone-chitosan derivatives prepared in Example 1

The preservative challenge results of the blank control cream (ie without the tropolone-chitosan derivative prepared in Example 1) are shown in Table 7.

Table 7: Antiseptic Challenge Results of Placebo Creams

The preservative challenge results of the essence containing the tropolone-chitosan derivative prepared in Example 1 are shown in Table 8.

Table 8: Antiseptic challenge results of the essence containing tropolone-chitosan derivatives prepared in Example 1

Table 9 shows the antiseptic challenge results of the blank control essence (that is, without the tropolone-chitosan derivative prepared in Example 1).

Table 9: Antiseptic challenge results of blank control essence

As can be seen from Table 6-9, in the antiseptic test of cream and essence to Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, Candida albicans, from the 7th day onwards, containing the preparation of Example 1 Bacteria and fungus in the sample of the tropolone-chitosan derivative obtained are all＜10cfu/g (or cfu/mL), illustrate the sample containing the tropolone-chitosan derivative that embodiment 1 makes The anti-corrosion system has a particularly strong inhibitory effect on microorganisms, which shows that the products added with the tropolone-chitosan derivatives of the present invention are not easily polluted by microorganisms during production, storage and use. Replacing the tropolone-chitosan derivative prepared in Example 1 with the tropolone-chitosan derivative prepared in Example 2-6 also has a similar antiseptic effect.

It can thus be proved that the tropolone-chitosan derivative of the present invention has better inhibitory and killing effects, broad spectrum and high efficiency, and has broad application prospects.

5. Application effect in oral daily products

Oral daily products mainly include toothpaste, mouthwash, tooth powder, etc. The application of tropolone-chitosan derivatives of the present invention in such oral daily products can not only solve the above-mentioned antiseptic problems, but also act on oral cavity Anaerobic bacteria, inhibit the reproduction of oral anaerobic bacteria in the oral cavity. As we all know, the most obvious symptom of oral anaerobic infection is bad breath after getting up in the morning and gradually aggravating, followed by symptoms such as dry mouth, pulpitis, periodontitis, gingivitis, nasal congestion or gingival swelling and pain. And the anaerobic bacteria that cause this series of problems include Fusobacterium nucleatum subsp. polynucleatum, Actinomyces viscosus, Actinobacillus actinomycetes, Streptococcus farus, Streptococcus mutans etc., the embodiment of the present invention 1-6 produces The obtained tropolone-chitosan derivatives all have strong inhibitory effects on the above oral anaerobic bacteria, and the antibacterial results are shown in Table 10 below.

Table 10: Minimum inhibitory concentration (MIC, unit ppm) of tropolone-chitosan derivatives to oral anaerobic bacteria

It can be seen from Table 10 that the tropolone-chitosan derivatives prepared by the present invention have a strong inhibitory effect on oral anaerobic bacteria, thereby effectively reducing the symptoms of oral anaerobic bacteria infection.

Claims (10)

1. A tolfenidone-chitosan derivative, which is characterized in that the structural formula of the tolfenidone-chitosan derivative is shown as a formula (1):

wherein R is ₁ Represents at least one of-H, hydroxyl-containing alkyl or carboxyl-containing alkyl;

R ₂ represents at least one of-H, hydroxyl-containing alkyl, carboxyl-containing alkyl or carbonyl-containing alkyl;

x and y each independently represent a positive integer;

the material containing the structural unit of the tolterone is replaced by mannitol.

2. The tolfenidone-chitosan derivative according to claim 1, wherein R ₁ Selected from-H, -CH ₂ COOH、-CH ₂ CH ₂ OH or-CH ₂ CH ₂ (OH)CH ₃ At least one of them.

3. The tolfenidone-chitosan derivative according to claim 1, wherein R ₂ Selected from-H, -COCH ₃ 、CH ₂ COOH or CH ₂ CH ₂ (OH)CH ₃ At least one of them.

4. A tolfenidone-chitosan derivative according to any of claims 1-3, wherein the raw material components for preparing the tolfenidone-chitosan derivative comprise chitosan or chitosan derivatives, and mannitol.

5. A process for the preparation of a tolfenidone-chitosan derivative according to any one of claims 1-4, characterized in that it comprises the steps of:

dissolving the chitosan or chitosan derivative in a solvent, then adding mannitol, and heating for reaction to obtain the tolphenol ketone-chitosan derivative.

6. The method according to claim 5, wherein the chitosan derivative is carboxybutyl chitosan.

7. The method according to claim 6, wherein the molecular weight of the carboxybutyl chitosan is 300000Da, the degree of deacetylation is 90%, the degree of O substitution is 75%, and the degree of N substitution is 6%.

8. The method of claim 5, wherein the temperature of the heating reaction is 55-90 ℃; the heating reaction time is 1-3 hours.

9. Use of a tolfenidone-chitosan derivative according to any one of claims 1-4 for the preparation of an antibacterial substance.

10. Use of the tolfenidone-chitosan derivative of any one of claims 1-4 in the medical, cosmetic or food field.