Medicine for treating sepsis-related encephalopathy and preparation method thereof

Abstract

The invention discloses a medicine for treating sepsis-related encephalopathy and a preparation method thereof, and the medicine is prepared from the following components in parts by weight: 0.7-0.9 part of 2-hydroxyethyl disulfide, 3.2-3.6 parts of triethylamine, 0.6-0.8 part of methacryloyl chloride, 3.1-3.5 parts of succinic anhydride, 3.7-3.9 parts of 4-dimethyl iminopyridine, 3.6-4.1 parts of mangiferin, 0.1-0.3 part of N, N-dimethylformamide, 0.1-0.3 part of sodium dodecyl benzene sulfonate and the balance of water. The composition is prepared from the following components in parts by weight: 2.9 to 3.3 parts of N, N '-dicyclohexylcarbodiimide, 2.1 to 2.5 parts of modified apigenin and 1.4 to 1.6 parts of salidroside. The absorption effect is improved by modifying the apigenin and preparing the nano-micelle load, and meanwhile, the apigenin is compounded with the salidroside to achieve synergistic interaction, so that the treatment effect is improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of sepsis, and specifically relates to a drug for treating sepsis-related encephalopathy and a preparation method thereof. Background Art

[0002] Sepsis is a life-threatening organ dysfunction syndrome caused by an imbalance in the body's stress response to infection. After sepsis occurs, various inflammatory cells in the body, including macrophages and endothelial cells, become activated, producing large amounts of free radicals, which in turn damage the function of various organs. Approximately 31 million patients are diagnosed with sepsis each year worldwide. It is the leading cause of hospitalization and death in intensive care units worldwide. Approximately 70% of sepsis patients develop brain dysfunction.

[0003] Sepsis-related encephalopathy is a serious complication of the central nervous system caused by sepsis. It is a diffuse brain dysfunction caused by non-intracranial infection, also known as septic brain injury. It has high mortality and disability rates, mainly characterized by impaired consciousness. It involves pathological processes such as changes in blood-brain barrier permeability, neuroinflammation, and oxidative stress. The main manifestations are altered consciousness such as delirium and coma, and include attention deficit, anxiety, abnormal behavior, disorientation, confusion, paralysis, drowsiness, coma, extensor rigidity, and peripheral nerve dysfunction.

[0004] Neuroinflammation is an important part of sepsis-related encephalopathy. The body produces a large amount of lipopolysaccharide that acts on vascular endothelial cells, activates microglia, and produces proinflammatory cytokines. For sepsis-related pathological inflammation, the use of antibiotics is still the main means; apigenin is a flavonoid compound extracted from celery. It is widely found in some vegetables, fruits, plant-based beverages, condiments and some medicinal plants, especially in celery. Apigenin can downregulate the expression of proinflammatory cytokines and adhesion molecules, but apigenin only exists in food in the form of glycosides, has low water solubility, and poor oral bioavailability. After ingestion, most apigenin is excreted or not absorbed, or is rapidly metabolized after absorption and does not show activity in the body.

[0005] The main problems with the current existing technologies are as follows: antibiotics have obvious toxic side effects, plant extracts are poorly absorbed and have poor efficacy, and there are no safe and effective therapeutic drugs. Summary of the Invention

[0006] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a drug for treating sepsis-related encephalopathy and a preparation method thereof. In order to solve the problem of poor absorption and poor efficacy of plant extracts, the present invention proposes to improve the absorption effect by modifying apigenin and preparing nanomicelle loading, and at the same time compound it with salidroside to achieve synergistic enhancement, thereby achieving improved therapeutic effect.

[0007] In order to achieve the above-mentioned purpose, the technical solution adopted by the present invention is as follows: The present invention provides a drug for treating sepsis-related encephalopathy, which is prepared from the following components in parts by weight: 0.7-0.9 parts of 2-hydroxyethyl disulfide, 3.2-3.6 parts of triethylamine, 0.6-0.8 parts of methacryloyl chloride, 3.1-3.5 parts of succinic anhydride, 3.7-3.9 parts of 4-dimethyliminopyridine, 3.6-4.1 parts of mangiferin, 2.9-3.3 parts of N,N'-dicyclohexylcarboximide, 2.1-2.5 parts of modified apigenin and 1.4-1.6 parts of salidroside.

[0008] Preferably, the modified apigenin is prepared from the following components in parts by weight: 2.5-2.7 parts of apigenin, 5.5-6.1 parts of triethylamine and 3.5-4.1 parts of propionyl chloride.

[0009] Preferably, the preparation method of the modified apigenin specifically comprises the following steps:

[0010] (1) Add apigenin to dichloromethane, add triethylamine under nitrogen protection, stir at 50 rpm in a 0°C ice-water bath for 30 min, add propionyl chloride, mix well, and stir in a water bath to obtain a reaction solution;

[0011] (2) Saturated ammonium chloride solution was added dropwise to the reaction solution obtained in step (1) to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine three times, dried with anhydrous sodium sulfate to remove moisture, and then concentrated under reduced pressure and purified to obtain modified apigenin.

[0012] Preferably, in step (1), the amount of apigenin added to dichloromethane is 25-30 mg / mL;

[0013] Preferably, in step (1), the water bath stirring temperature is 15-20°C, the speed is 60-80 rpm, and the time is 1-2 h.

[0014] Preferably, in step (2), the volume of the ammonium chloride solution is 0.5-0.6 times the volume of the reaction solution obtained in step (1).

[0015] The present invention also provides a method for preparing a drug for treating sepsis-related encephalopathy, which specifically comprises the following steps:

[0016] S1. Add 2-hydroxyethyl disulfide to tetrahydrofuran, add triethylamine, stir in a water bath, add dropwise 0.25 wt% methacryloyl chloride, raise the temperature to 20°C, continue stirring for 24 h, then wash with petroleum ether and purify to obtain an intermediate product;

[0017] S2. Add the intermediate product obtained in S1 to tetrahydrofuran, then add succinic anhydride and 4-dimethyliminopyridine, mix well, stir in a water bath, add 0.1 times the volume of deionized water to stop the reaction, extract with dichloromethane and saturated sodium chloride solution in sequence, collect the organic layer, dry, remove the solvent by rotary evaporation, wash with petroleum ether, and purify to obtain a carboxyl derivative;

[0018] S3. The carboxyl derivative obtained in S2 was added to dichloromethane at an addition amount of 18 mg / mL, followed by the addition of mangiferin and N,N'-dicyclohexylcarboximide, and stirred at room temperature for 24 h. The mixture was extracted, dried, and evaporated to remove the solvent to obtain a mixture. The mixture was added to dimethyl sulfoxide at an addition amount of 90 mg / mL, and 1.8% of azobisisobutyronitrile by weight of the mixture was added. The mixture was reacted at 80°C under nitrogen protection for 6 h. The solution was then cooled to room temperature, and 9 times the volume of methyl tert-butyl ether was added for precipitation. The mixture was filtered, dialyzed, and dried to obtain nanomicelles.

[0019] S4. The nanomicelles obtained in S3 were added to dimethyl sulfoxide, modified apigenin and salidroside were added, and the reaction was carried out in a light-locked state for 3 hours. Subsequently, 3 times the volume of deionized water was added and stirred for 20 hours. Subsequently, the mixture was dialyzed with deionized water for 20 hours, and finally freeze-dried to obtain a drug for treating sepsis-related encephalopathy.

[0020] Preferably, in S1, the amount of 2-hydroxyethyl disulfide added to tetrahydrofuran is 0.7-0.9 mg / mL;

[0021] Preferably, in S1, the stirring temperature of the water bath is 0-3°C, the speed is 60-100 rpm, and the time is 30-40 min.

[0022] Preferably, in S2, the amount of the intermediate product obtained in S1 added to tetrahydrofuran is 0.11-0.13 g / mL;

[0023] Preferably, in S2, the water bath stirring temperature is 20-25°C, the speed is 80-100 rpm, and the time is 15-20 h.

[0024] Preferably, in S4, the amount of the nanomicelles obtained in S3 added to dimethyl sulfoxide is 8-10 mg / mL.

[0025] The beneficial effects achieved by the present invention are as follows: the chemical structure of apigenin is modified by propionyl chloride, an acetyl group is introduced, the lipid-water partition coefficient of the compound in the body is improved, the dissolution and absorption of apigenin in the body are enhanced, the lipophilicity of the apigenin derivative is significantly increased, and the penetration ability of the apigenin to the internal membranes and tissues of the organism is improved, thereby increasing the absorption degree and transfer speed of the modified apigenin in the organism; further, a polymer with a disulfide bond is prepared by 2-hydroxyethyl disulfide, and the modified apigenin and salidroside are loaded by self-assembly to obtain drug-loaded micelles with active oxygen response ability and targeted release ability. Mangiferin has anti-inflammatory and neuroprotective effects, apigenin has multiple functions such as anti-inflammatory, anti-oxidative stress, and anti-apoptosis, which can significantly inhibit the production of reactive oxygen species and down-regulate the expression of pro-inflammatory cytokines and adhesion molecules. Salidroside has anti-inflammatory, anti-hypoxia, blocking inflammatory pathways, brain-boosting and blood-activating effects, and can reduce the concentration of S100β protein in the patient's serum and cerebrospinal fluid, thereby improving the patient's intelligence, memory and living ability. The three are used together to synergistically enhance the effect and improve the recovery rate of the mobility and cognitive abilities of patients with sepsis-related encephalopathy. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The results of the cytotoxicity tests of Examples 1-3 and Comparative Example 1 are shown;

[0027] Figure 2 The results of the response test of Examples 1-3 are shown;

[0028] Figure 3 The results of the mouse tests of Examples 1-3 and Comparative Examples 1-3 are shown;

[0029] Figure 4 This is a flow chart of preparing modified apigenin according to Example 1 of the present invention.

[0030] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention. DETAILED DESCRIPTION

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0032] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the present invention. The preferred embodiments and materials described herein are for illustrative purposes only and are not intended to limit the scope of this application.

[0033] The experimental methods in the following examples are conventional methods unless otherwise specified; the experimental materials used in the following examples are purchased from commercial channels unless otherwise specified.

[0034] Example 1

[0035] A drug for treating sepsis-related encephalopathy is prepared from the following components in parts by weight: 0.7 parts of 2-hydroxyethyl disulfide, 3.2 parts of triethylamine, 0.6 parts of methacryloyl chloride, 3.1 parts of succinic anhydride, 3.7 parts of 4-dimethyliminopyridine, 3.6 parts of mangiferin, 2.9 parts of N,N'-dicyclohexylcarboximide, 2.1 parts of modified apigenin, and 1.4 parts of salidroside.

[0036] The modified apigenin was prepared from the following components in parts by weight: 2.5 parts of apigenin, 5.5 parts of triethylamine and 3.5 parts of propionyl chloride.

[0037] The preparation method of modified apigenin specifically comprises the following steps:

[0038] (1) Apigenin was added to dichloromethane at a dosage of 25 mg / mL, triethylamine was added under nitrogen protection, and the mixture was stirred at 50 rpm in a 0°C ice-water bath for 30 min. Propionyl chloride was added and mixed evenly, and the mixture was stirred at 60 rpm in a 15°C water bath for 1 h to obtain a reaction solution;

[0039] (2) 0.5 times the volume of saturated ammonium chloride solution was added dropwise to the reaction solution obtained in step (1) to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine three times, dried with anhydrous sodium sulfate to remove moisture, and then concentrated under reduced pressure and purified to obtain modified apigenin.

[0040] The present invention also provides a method for preparing a drug for treating sepsis-related encephalopathy, which specifically comprises the following steps:

[0041] S1. 2-Hydroxyethyl disulfide was added to tetrahydrofuran at a dosage of 0.7 mg / mL, triethylamine was added, and the mixture was stirred at 60 rpm in a 0°C water bath for 30 min. 0.25 wt% methacryloyl chloride was added dropwise, and the mixture was heated to 20°C and stirred for 24 h. The mixture was then washed with petroleum ether and purified to obtain an intermediate product.

[0042] S2. The intermediate product obtained in S1 was added to tetrahydrofuran at an addition amount of 0.11 g / mL, followed by the addition of succinic anhydride and 4-dimethyliminopyridine, mixed evenly, stirred in a 20°C water bath at 80 rpm for 15 h, and 0.1 times the volume of deionized water was added to stop the reaction. The mixture was extracted with dichloromethane and saturated sodium chloride solution in sequence, the organic layer was collected, dried, and the solvent was removed by rotary evaporation, washed with petroleum ether, and purified to obtain a carboxyl derivative;

[0043] S3. The carboxyl derivative obtained in S2 was added to dichloromethane at an addition amount of 18 mg / mL, followed by the addition of mangiferin and N,N'-dicyclohexylcarboximide, and stirred at room temperature for 24 h. The mixture was extracted, dried, and evaporated to remove the solvent to obtain a mixture. The mixture was added to dimethyl sulfoxide at an addition amount of 90 mg / mL, and 1.8% of azobisisobutyronitrile by weight of the mixture was added. The mixture was reacted at 80°C under nitrogen protection for 6 h. The solution was then cooled to room temperature, and 9 times the volume of methyl tert-butyl ether was added for precipitation. The mixture was filtered, dialyzed, and dried to obtain nanomicelles.

[0044] S4. The nanomicelles obtained in S3 were added to dimethyl sulfoxide at an addition amount of 8 mg / mL, and modified apigenin and salidroside were added. The reaction was blocked from light for 3 hours, and then 3 times the volume of deionized water was added and stirred for 20 hours. The mixture was then dialyzed with deionized water for 20 hours, and finally freeze-dried to obtain a drug for treating sepsis-related encephalopathy.

[0045] Example 2

[0046] A drug for treating sepsis-related encephalopathy is prepared from the following components in parts by weight: 0.9 parts of 2-hydroxyethyl disulfide, 3.6 parts of triethylamine, 0.8 parts of methacryloyl chloride, 3.5 parts of succinic anhydride, 3.9 parts of 4-dimethyliminopyridine, 4.1 parts of mangiferin, 3.3 parts of N,N'-dicyclohexylcarboximide, 2.5 parts of modified apigenin, and 1.6 parts of salidroside.

[0047] The modified apigenin was prepared from the following components in parts by weight: 2.7 parts of apigenin, 6.1 parts of triethylamine and 4.1 parts of propionyl chloride.

[0048] The preparation method of modified apigenin specifically comprises the following steps:

[0049] (1) Apigenin was added to dichloromethane at a dosage of 30 mg / mL, triethylamine was added under nitrogen protection, and the mixture was stirred at 50 rpm in a 0°C ice-water bath for 30 min. Propionyl chloride was added and mixed evenly, and the mixture was stirred at 80 rpm in a 20°C water bath for 2 h to obtain a reaction solution;

[0050] (2) 0.6 times the volume of saturated ammonium chloride solution was added dropwise to the reaction solution obtained in step (1) to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine three times, dried with anhydrous sodium sulfate to remove moisture, and then concentrated under reduced pressure and purified to obtain modified apigenin.

[0051] The present invention also provides a method for preparing a drug for treating sepsis-related encephalopathy, which specifically comprises the following steps:

[0052] S1. 2-Hydroxyethyl disulfide was added to tetrahydrofuran at a concentration of 0.9 mg / mL, triethylamine was added, and the mixture was stirred at 100 rpm in a 3°C water bath for 40 min. 0.25 wt% methacryloyl chloride was added dropwise, and the mixture was heated to 20°C and stirred for 24 h. The mixture was then washed with petroleum ether and purified to obtain an intermediate product.

[0053] S2. The intermediate product obtained in S1 was added to tetrahydrofuran at an addition amount of 0.13 g / mL, followed by the addition of succinic anhydride and 4-dimethyliminopyridine, mixed evenly, and stirred in a 25°C water bath at 100 rpm for 20 h. 0.1 times the volume of deionized water was added to stop the reaction, and the mixture was extracted with dichloromethane and saturated sodium chloride solution in sequence. The organic layer was collected, dried, and the solvent was removed by rotary evaporation. The mixture was washed with petroleum ether and purified to obtain a carboxyl derivative.

[0054] S3. The carboxyl derivative obtained in S2 was added to dichloromethane at an addition amount of 18 mg / mL, followed by the addition of mangiferin and N,N'-dicyclohexylcarboximide, and stirred at room temperature for 24 h. The mixture was extracted, dried, and evaporated to remove the solvent to obtain a mixture. The mixture was added to dimethyl sulfoxide at an addition amount of 90 mg / mL, and 1.8% of azobisisobutyronitrile by weight of the mixture was added. The mixture was reacted at 80°C under nitrogen protection for 6 h. The solution was then cooled to room temperature, and 9 times the volume of methyl tert-butyl ether was added for precipitation. The mixture was filtered, dialyzed, and dried to obtain nanomicelles.

[0055] S4. The nanomicelles obtained in S3 were added to dimethyl sulfoxide at a dosage of 10 mg / mL, and modified apigenin and salidroside were added. The reaction was carried out in a light-locked state for 3 hours, and then 3 times the volume of deionized water was added and stirred for 20 hours. The mixture was then dialyzed with deionized water for 20 hours, and finally freeze-dried to obtain a drug for treating sepsis-related encephalopathy.

[0056] Example 3

[0057] A drug for treating sepsis-related encephalopathy is prepared from the following components in parts by weight: 0.8 parts of 2-hydroxyethyl disulfide, 3.4 parts of triethylamine, 0.7 parts of methacryloyl chloride, 3.3 parts of succinic anhydride, 3.8 parts of 4-dimethyliminopyridine, 3.9 parts of mangiferin, 3.1 parts of N,N'-dicyclohexylcarboximide, 2.4 parts of modified apigenin, and 1.5 parts of salidroside.

[0058] The modified apigenin was prepared from the following components in parts by weight: 2.6 parts of apigenin, 5.9 parts of triethylamine and 3.8 parts of propionyl chloride.

[0059] The preparation method of modified apigenin specifically comprises the following steps:

[0060] (1) Apigenin was added to dichloromethane at a dosage of 27 mg / mL, triethylamine was added under nitrogen protection, and the mixture was stirred at 50 rpm in a 0°C ice-water bath for 30 min. Propionyl chloride was added and mixed evenly, and the mixture was stirred at 70 rpm in a 18°C ​​water bath for 1.5 h to obtain a reaction solution;

[0061] (2) 0.55 times the volume of saturated ammonium chloride solution was added dropwise to the reaction solution obtained in step (1) to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine three times, dried with anhydrous sodium sulfate to remove moisture, and then concentrated under reduced pressure and purified to obtain modified apigenin.

[0062] The present invention also provides a method for preparing a drug for treating sepsis-related encephalopathy, which specifically comprises the following steps:

[0063] S1. 2-Hydroxyethyl disulfide was added to tetrahydrofuran at a dosage of 0.8 mg / mL, triethylamine was added, and the mixture was stirred at 80 rpm in a 2°C water bath for 35 min. 0.25 wt% methacryloyl chloride was added dropwise, and the mixture was heated to 20°C and stirred for 24 h. The mixture was then washed with petroleum ether and purified to obtain an intermediate product.

[0064] S2. The intermediate product obtained in S1 was added to tetrahydrofuran at an addition amount of 0.12 g / mL, followed by the addition of succinic anhydride and 4-dimethyliminopyridine, mixed evenly, and stirred in a 22°C water bath at 90 rpm for 18 h. 0.1 times the volume of deionized water was added to stop the reaction, and the mixture was extracted with dichloromethane and saturated sodium chloride solution in sequence. The organic layer was collected, dried, and the solvent was removed by rotary evaporation. The mixture was washed with petroleum ether and purified to obtain a carboxyl derivative.

[0065] S3. The carboxyl derivative obtained in S2 was added to dichloromethane at an addition amount of 18 mg / mL, followed by the addition of mangiferin and N,N'-dicyclohexylcarboximide, and stirred at room temperature for 24 h. The mixture was extracted, dried, and evaporated to remove the solvent to obtain a mixture. The mixture was added to dimethyl sulfoxide at an addition amount of 90 mg / mL, and 1.8% of azobisisobutyronitrile by weight of the mixture was added. The mixture was reacted at 80°C under nitrogen protection for 6 h. The solution was then cooled to room temperature, and 9 times the volume of methyl tert-butyl ether was added for precipitation. The mixture was filtered, dialyzed, and dried to obtain nanomicelles.

[0066] S4. The nanomicelles obtained in S3 were added to dimethyl sulfoxide at a dosage of 9 mg / mL, modified apigenin and salidroside were added, and the reaction was carried out in a light-locked state for 3 hours. Subsequently, 3 times the volume of deionized water was added and stirred for 20 hours. Subsequently, the mixture was dialyzed with deionized water for 20 hours, and finally freeze-dried to obtain a drug for treating sepsis-related encephalopathy.

[0067] Comparative Example 1

[0068] This comparative example provides a drug, which differs from Example 1 only in that the drug does not contain nanomicelles, and the remaining components and component contents are the same as those in Example 1.

[0069] Comparative Example 2

[0070] This comparative example provides a medicine, which differs from Example 1 only in that apigenin is used instead of modified apigenin in the components, and the other components and component contents are the same as those in Example 1.

[0071] Comparative Example 3

[0072] This comparative example provides a medicine, which differs from Example 1 only in that salidroside is replaced by apigenin in the components, and the other components and component contents are the same as those in Example 1.

[0073] Experimental example

[0074] 1. Cytotoxicity Test

[0075] Zebrafish embryos fertilized for 3 hours were randomly divided and transferred to 24-well plates, with 50 eggs per well. The volume of the culture medium was 2 mL. Examples 1-3 and Comparative Example 1 were added to form the experimental group, and the concentration of salidroside reached 100 μg / mL. An equal concentration of physiological saline was added to form the blank group. Three wells were set up in each group. The embryos were treated and cultured in a constant temperature and humidity incubator at 28.5° C. for 3 days. The survival rate of the eggs on the third day was then calculated using the following formula:

[0076] Survival rate = number of surviving eggs in the experimental group / number of surviving eggs in the blank group × 100%.

[0077] Figure 1 The results of the cytotoxicity test of Examples 1-3 and Comparative Example 1 are shown in the figure. As shown in the figure, after 3 days, the survival rates of Examples 1-3 and Comparative Example 1 are 91%, 93%, 92%, and 63%, respectively; the survival rate of Comparative Example 1 is significantly lower than that of Examples 1-3, indicating that the preparation of nanomicelles reduces the toxicity of high-concentration drugs to cells.

[0078] 2. Response Testing

[0079] 5 mg of each of Examples 1-3 was added to 5 mL of 0.1% H2O2 and deionized water, and quickly transferred to a dialysis bag and immersed in 50 mL of deionized water. The mixture was shaken at 37°C and 100 rpm. After 24 h, the release amount of salidroside in deionized water was measured by spectrophotometry, and the release rate was calculated using the following formula:

[0080] Release rate = release amount / initial drug loading × 100%.

[0081] Figure 2 This is the result of the response test of Examples 1-3. As shown in the figure, after 24 hours, the release rates of Examples 1-3 in H2O2 solution and deionized water are 65.9%, 66.3%, 66.1% and 21.6%, 22.5%, 21.9%, respectively; the release rate of Examples 1-3 in H2O2 solution is significantly higher than that in deionized water, and the prepared drugs have good responsiveness.

[0082] 3. Mouse Testing

[0083] Ten-week-old C57BL / 6 wild-type mice were selected, and the cecum was ligated with a No. 3 suture at a position approximately 1 / 2 of the length of the cecum. After ligation, a No. 8 needle was used to penetrate the end of the cecum twice, a small amount of intestinal contents was squeezed out at the needle hole, and the cecum was returned to the abdominal cavity to prepare a sepsis mouse model. The mice were randomly divided into groups, with 50 mice in each group, and treated with Examples 1-3, Comparative Examples 1-3, and normal saline intravenous injection, respectively. The neurobehavioral changes of the mice were evaluated. The evaluation content included five contents: escape reflex, corneal reflex, righting reflex, auricular reflex, and tail flick reflex. 0 was considered to be no reflex, 1 was considered to be a weakened reflex (if no reflex appeared within 10 seconds), and 2 was considered to be normal reflex. A neurobehavioral reflex score of ≤6 was recorded as a neurobehavioral change, and the neurobehavioral change rate of each group was calculated using the following formula:

[0084] The rate of neurobehavioral changes = the number of mice with neurobehavioral changes / the total number of mice × 100%.

[0085] Figure 3 The results of the test on mice in Example 1-3, Comparative Example 1-3 and the normal saline group are shown in the figure. As shown in the figure, after 3 weeks, the neurobehavioral change rates of Example 1-3, Comparative Example 1-3 and the normal saline group were 12%, 14%, 10%, 43%, 51%, 56% and 76%, respectively; the neurobehavioral change rate of Example 1-3 was significantly lower than that of Comparative Example 1, indicating that the preparation of nanomicelles effectively reduced the neurobehavioral change rate, the neurobehavioral change rate of Example 1-3 was significantly lower than that of Comparative Example 2, indicating that the modification of apigenin effectively reduced the neurobehavioral change rate, the neurobehavioral change rate of Example 1-3 was significantly lower than that of Comparative Example 3, indicating that the use of salidroside improved the therapeutic effect of the drug.

[0086] Figure 4 This is a flow chart of preparing modified apigenin in Example 1 of the present invention. As shown in the figure, the modified apigenin is obtained by modifying the chemical structure of apigenin with propionyl chloride.

[0087] While the embodiments of the present invention have been shown and described, it will be apparent to those skilled in the art that various changes, modifications, substitutions, and alterations can be made to the embodiments without departing from the principles and spirit of the invention.

[0088] The present invention and its embodiments are described above. Such description is not restrictive. The drawings show only one embodiment of the present invention, and actual applications are not limited thereto. In short, if a person skilled in the art is inspired by the above, and does not deviate from the purpose of the present invention, any method and embodiment similar to the technical solution without creative design shall fall within the scope of protection of the present invention.

Claims

1. A drug for treating sepsis-related encephalopathy, characterized in that: Prepared from the following components in parts by weight: 0.7-0.9 parts of 2-hydroxyethyl disulfide, 3.2-3.6 parts of triethylamine, 0.6-0.8 parts of methacryloyl chloride, 3.1-3.5 parts of succinic anhydride, 3.7-3.9 parts of 4-dimethyliminopyridine, 3.6-4.1 parts of mangiferin, 2.9-3.3 parts of N,N'-dicyclohexylcarboximide, 2.1-2.5 parts of modified apigenin and 1.4-1.6 parts of salidroside; The modified apigenin is prepared from the following components in parts by weight: 2.5-2.7 parts of apigenin, 5.5-6.1 parts of triethylamine and 3.5-4.1 parts of propionyl chloride.

2. A method for preparing the drug for treating sepsis-related encephalopathy according to claim 1, characterized in that: The specific steps include: S1. Add 2-hydroxyethyl disulfide to tetrahydrofuran, add triethylamine, stir in a water bath, add dropwise 0.25 wt% methacryloyl chloride, raise the temperature to 20°C, continue stirring for 24 h, then wash with petroleum ether and purify to obtain an intermediate product; S2. Add the intermediate product obtained in S1 to tetrahydrofuran, then add succinic anhydride and 4-dimethyliminopyridine, mix well, stir in a water bath, add 0.1 times the volume of deionized water to stop the reaction, extract with dichloromethane and saturated sodium chloride solution in sequence, collect the organic layer, dry, remove the solvent by rotary evaporation, wash with petroleum ether, and purify to obtain a carboxyl derivative; S3. The carboxyl derivative obtained in S2 was added to dichloromethane at an addition amount of 18 mg / mL, followed by the addition of mangiferin and N,N'-dicyclohexylcarboximide, and stirred at room temperature for 24 h. The mixture was extracted, dried, and evaporated to remove the solvent to obtain a mixture. The mixture was added to dimethyl sulfoxide at an addition amount of 90 mg / mL, and 1.8% of azobisisobutyronitrile by weight of the mixture was added. The mixture was reacted at 80°C under nitrogen protection for 6 h. The solution was then cooled to room temperature, and 9 times the volume of methyl tert-butyl ether was added for precipitation. The mixture was filtered, dialyzed, and dried to obtain nanomicelles. S4. The nanomicelles obtained in S3 were added to dimethyl sulfoxide, modified apigenin and salidroside were added, and the reaction was carried out in a light-locked state for 3 hours. Subsequently, 3 times the volume of deionized water was added and stirred for 20 hours. Subsequently, the mixture was dialyzed with deionized water for 20 hours, and finally freeze-dried to obtain a drug for treating sepsis-related encephalopathy.

3. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 2, characterized in that: The preparation method of the modified apigenin specifically comprises the following steps: (1) Add apigenin to dichloromethane, add triethylamine under nitrogen protection, stir at 50 rpm in a 0°C ice-water bath for 30 min, add propionyl chloride, mix well, and stir in a water bath to obtain a reaction solution; (2) Saturated ammonium chloride solution was added dropwise to the reaction solution obtained in step (1) to quench the reaction, and the mixture was extracted with ethyl acetate three times. The organic phases were combined, washed with saturated brine three times, dried with anhydrous sodium sulfate to remove moisture, and then concentrated under reduced pressure and purified to obtain modified apigenin.

4. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 3, characterized in that: In step (1), the amount of apigenin added to dichloromethane is 25-30 mg / mL; the water bath stirring temperature is 15-20°C, the speed is 60-80 rpm, and the time is 1-2 h.

5. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 4, characterized in that: In step (2), the volume of the ammonium chloride solution is 0.5-0.6 times that of the reaction solution obtained in step (1).

6. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 2, characterized in that: In S1, the amount of 2-hydroxyethyl disulfide added to tetrahydrofuran is 0.7-0.9 mg / mL; the water bath stirring temperature is 0-3°C, the speed is 60-100 rpm, and the time is 30-40 min.

7. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 6, characterized in that: In S2, the intermediate product obtained in S1 is added to tetrahydrofuran in an amount of 0.11-0.13 g / mL; the water bath stirring temperature is 20-25°C, the speed is 80-100 rpm, and the time is 15-20 h.

8. The method for preparing the drug for treating sepsis-related encephalopathy according to claim 7, characterized in that: In S4, the nanomicelles obtained in S3 were added to dimethyl sulfoxide in an amount of 8-10 mg / mL.

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