CN117771212A - Resina Draconis perchlorate solid lipid nanoparticle, and preparation method and application thereof - Google Patents

Abstract

The invention provides a preparation method of solid lipid nanoparticles of resina Draconis perchlorate, which comprises the following steps: a) Mixing glyceryl monostearate, soybean phospholipid and resina Draconis perchlorate, and dissolving with solvent to obtain organic phase; b) An aqueous solution of poloxamer as the aqueous phase; c) The organic phase is added into the water phase and cooled to obtain the product. According to the invention, the glyceryl monostearate, the soybean lecithin and the poloxamer are used as coating materials to coat the resina Draconis perchlorate, so that the finally prepared resina Draconis perchlorate solid lipid nanoparticle has high stability, high encapsulation efficiency and high drug loading.

Description

Hematocritin perchlorate solid lipid nanoparticles and preparation methods and applications thereof

Technical field

The present invention relates to the technical field of pharmaceutical preparations, and in particular to a kind of hematocritin perchlorate solid lipid nanoparticles and preparation methods and applications thereof.

Background technique

As the main biologically active component of Dracula, it is a flavonoid compound and is an indicator compound for quality control of Dracula. However, its chemical properties are unstable and it is easily decomposed by external factors such as light, temperature and pH value. As a synthetic analog of hematocritin, hematocritin perchlorate is a standard for measuring hematocritin content stipulated in the "Pharmacopoeia of the People's Republic of China (2020 Edition)". Studies have reported that it has anti-tumor properties, promotes wound healing, and prevents and treats diabetes. , anti-leukemia, anti-ulcer, anti-inflammatory and other effects.

Therefore, it is very necessary to develop a hematocritin perchlorate solid lipid nanoparticle with high stability, encapsulation efficiency and drug loading capacity.

Contents of the invention

In view of this, the technical problem to be solved by the present invention is to provide a dracotoxin perchlorate solid lipid nanoparticle. The nanoparticle provided by the present invention has high stability, high encapsulation efficiency and high drug loading capacity.

The invention provides a method for preparing hedradin perchlorate solid lipid nanoparticles, which includes:

A) glyceryl monostearate, soybean lecithin and dracoside perchlorate are mixed and dissolved in a solvent to obtain an organic phase;

B) The aqueous solution of poloxamer serves as the water phase;

C) Add the organic phase to the water phase and cool to obtain.

Preferably, the mass ratio of the glyceryl monostearate and soybean lecithin is 1: (1-3);

The mass ratio of the monostearate glyceryl, soybean lecithin and dracoside perchlorate is (8-12):(18-22):(1-3).

Preferably, the concentration of the poloxamer aqueous solution is 10-40 mg/mL.

Preferably, the mass ratio of the glyceryl monostearate, soybean lecithin and poloxamer is (8-12): (18-22): (10-14).

Preferably, the solvent in step A) is ethanol; the dissolution is ultrasonic dissolution, and the power of the ultrasonic is 200-300 Hz; the temperature of the dissolution is 70-80°C;

Step B) the aqueous solution of poloxamer is obtained by dissolving poloxamer in water; the dissolution temperature is 70-80°C.

Preferably, the specific method of adding the organic phase to the water phase in step C) is: adding the organic phase to the water by slow dripping at 70-80°C and stirring, and stirring at 80°C for 20-30 minutes; the stirring speed is: 1000～1400r/min;

The cooling is specifically: cooling in ice water at 0 to 2°C, stirring for 50 to 70 minutes, and then continuing to cool at 25 to 30°C.

The invention provides a kind of hematocritin perchlorate solid lipid nanoparticles, which is prepared by the preparation method described in any one of the above technical solutions.

The present invention provides a hematocritin perchlorate drug, including the hematocritin perchlorate solid lipid nanoparticles described in the above technical solution.

Preferably, the dosage form of the drug is one or more of powder, suspension, tablet, capsule, granule or powder.

The present invention provides the application of the above-mentioned hedradin perchlorate solid lipid nanoparticles in the preparation of medicines for treating injurious neuralgia.

Preferably, the treatment includes reducing the expression of NMDAR 1 and/or increasing the pain threshold.

Compared with the existing technology, the present invention provides a method for preparing hedradin perchlorate solid lipid nanoparticles, including: A) glyceryl monostearate, soybean lecithin and hemodrin perchlorate Mix and dissolve with a solvent to obtain the organic phase; B) the aqueous solution of poloxamer is used as the water phase; C) the organic phase is added to the water phase and cooled to obtain. In the present invention, the above-mentioned glyceryl monostearate, soybean lecithin and poloxamer are used as coating materials to coat the above-mentioned hematocrit perchlorate, so that the finally prepared hematocritin perchlorate solid lipid nanoparticles are stable. High reactivity, high encapsulation efficiency and drug loading capacity.

Description of drawings

Fig. 1 is a diagram of dracoside perchlorate standard;

Figure 2 is the standard curve of hematocritin perchlorate;

Figure 3 Star point effect and contour map;

Figure 4 SLN particle size chart;

Figure 5: Effect of tincture on pathological changes of rat sciatic nerve (HE staining, 200×);

Figure 6 shows the in vitro drug release curves of hedradin original drug and hedradin emulsion.

Detailed ways

The present invention provides a hematocritin perchlorate solid lipid nanoparticle and its preparation method and application. Those skilled in the art can learn from the content of this article and appropriately improve the process parameters to achieve it. It should be pointed out in particular that all similar substitutions and modifications are obvious to those skilled in the art, and they all fall within the protection scope of the present invention. The methods and applications of the present invention have been described through preferred embodiments. Relevant persons can obviously modify or appropriately change and combine the methods and applications herein without departing from the content, spirit and scope of the present invention to implement and apply the present invention. Invent technology.

The present invention provides a method for preparing dracoside perchlorate solid lipid nanoparticles, comprising:

A) glyceryl monostearate, soybean lecithin and dracoside perchlorate are mixed and dissolved in a solvent to obtain an organic phase;

B) The aqueous solution of poloxamer serves as the water phase;

C) Add the organic phase into the aqueous phase and cool to obtain the product.

The preparation method of the hematocritin perchlorate solid lipid nanoparticles of the present invention first mixes glycerol monostearate, soybean lecithin and hematocritin perchlorate;

In some preferred embodiments of the present invention, the mass ratio of glyceryl monostearate to soybean lecithin is 1:(1-3);

Specifically, the mass ratio of glyceryl monostearate and soybean lecithin is 1:1, 1:2 or 1:3; or a point value between any two of the above.

In one of the preferred embodiments of the present invention, the mass ratio of the glyceryl monostearate, soybean lecithin and hematocritin perchlorate is preferably (8 to 12): (18 to 22): (1 to 3 );

In some preferred embodiments of the present invention, the mass ratio of glyceryl monostearate, soybean lecithin and dracotoxin perchlorate is preferably (9-11): (19-21): (1.1-2.8);

In one of the preferred embodiments of the present invention, the mass ratio of the glyceryl monostearate, soybean lecithin and hematocritin perchlorate is preferably 10:20:2.

In the present invention, a solvent is used to dissolve after mixing to obtain an organic phase; the solvent includes but is not limited to ethanol; the dissolution is ultrasonic dissolution, and the power of the ultrasonic is 200 to 300 Hz; the temperature of the dissolution is 70 to 80°C; more Preferably, the dissolution temperature is 80°C.

An aqueous solution of poloxamer served as the aqueous phase. The aqueous solution of poloxamer is specifically obtained by dissolving poloxamer in water; the dissolution temperature is 70-80°C; more preferably, the dissolution temperature is 80°C.

In certain embodiments, the concentration of the poloxamer aqueous solution is 10 to 40 mg/mL;

In certain specific embodiments, the concentration of the aqueous solution of poloxamer is 15-35 mg/mL.

In a specific embodiment, the concentration of the poloxamer aqueous solution is 20-30 mg/mL.

In some specific embodiments of the present invention, the mass ratio of the glyceryl monostearate, soybean lecithin and poloxamer is (8-12): (18-22): (10-14).

In some specific embodiments of the present invention, the mass ratio of the glyceryl monostearate, soybean lecithin and poloxamer is (9-11): (19-21): (11-13).

In some specific embodiments of the present invention, the mass ratio of the glyceryl monostearate, soybean lecithin and poloxamer is 10:20:12.

After obtaining the organic phase and the aqueous phase respectively, the organic phase is added to the aqueous phase. The above steps are preferably specifically as follows: under the condition of 70-80°C and stirring, the organic phase is slowly added to the water by dripping, and stirred at 80°C for 20-30 minutes; the stirring speed is 1000-1400r/min; more preferably 1000-1300r/min; most preferably 1000-1200r/min.

In some specific embodiments, the above steps are preferably specifically as follows: adding the organic phase to water by slow dripping at 75-80° C. and stirring at 80° C. for 25-30 min;

In some specific embodiments, the above steps are preferably as follows: at 80°C, under stirring conditions, the organic phase is slowly added to water by dripping, and stirred at 80°C for 30 minutes.

The above-mentioned slow drip specifically means that the infusion speed is controlled at 60-80 drops/minute.

The organic phase is added to the aqueous phase and cooled to obtain the product; the cooling is specifically: cooling in 0-2°C ice water, stirring for 50-70 minutes, and then continuing to cool at 25-30°C. More preferably, it is specifically: cooling in 0-2°C ice water, stirring for 60 minutes, and then continuing to cool at 25-30°C.

The invention provides a kind of hematocritin perchlorate solid lipid nanoparticles, which is prepared by the preparation method described in any one of the above technical solutions.

The above preparation method has been clearly described in the present invention and will not be described in detail here.

The present invention provides a hematocritin perchlorate medicine, including the hematocritin perchlorate solid lipid nanoparticles described in the above technical solution.

The drug of the present invention includes the dracotin perchlorate solid lipid nanoparticles described in the above technical solution, and also includes conventional excipients in the pharmaceutical field well known to those skilled in the art, and the inventors do not limit this.

The dosage form of the drug includes but is not limited to one or more of powder, suspension, tablet, capsule, granule or powder.

The present invention preferably prepares the above-mentioned drug into a lyophilized powder.

The present invention also provides a method for preparing dracotin perchlorate freeze-dried powder: the dracotin perchlorate solid lipid nanoparticles described in the above technical solution are mixed with a glucose solution, pre-frozen, and then freeze-dried to obtain the lyophilized powder.

Among them, the pre-freezing parameter is -80°C for 12 hours; the freeze-drying parameter is specifically: -80°C freeze-drying for 10 to 12 hours.

The concentration of the glucose solution is preferably 9% glucose solution.

The present invention provides the application of the above-mentioned hedradin perchlorate solid lipid nanoparticles in the preparation of medicines for treating injurious neuralgia.

Wherein, the treatment includes reducing the expression of NMDAR 1 and/or increasing the pain threshold.

The present invention also provides a method for treating traumatic neuralgia, which includes administering the above-mentioned hedradin perchlorate drug. Preferably, the method of administering the drug is oral administration.

The invention provides a method for preparing hemodacin perchlorate solid lipid nanoparticles, which includes: A) mixing glycerol monostearate, soybean lecithin and hemodacin perchlorate, and dissolving them with a solvent to obtain The organic phase; B) the aqueous solution of poloxamer is used as the water phase; C) the organic phase is added to the water phase and cooled to obtain. In the present invention, the above-mentioned glyceryl monostearate, soybean lecithin and poloxamer are used as coating materials to coat the above-mentioned hematocrit perchlorate, so that the finally prepared hematocritin perchlorate solid lipid nanoparticles are stable. High reactivity, high encapsulation efficiency and drug loading capacity.

In order to further illustrate the present invention, the hedradin perchlorate solid lipid nanoparticles provided by the present invention and its preparation method and application are described in detail below in conjunction with the examples.

Example 1 Establishment of a method for determining the content of dracoside perchlorate

1.1 Chromatographic conditions

Chromatographic conditions: Chromatographic column: Agilent ZORBAX SB-C18 (5μm, 4.6×250mm); mobile phase: acetonitrile-pure water aqueous solution;

Isocratic elution: 0-10min, 80% acetonitrile; 20% pure water;

Flow rate: 1mL·min-1; injection volume: 10μL; sample chamber temperature: 4℃; column temperature: 40℃;

Detection wavelength: Hedradin perchlorate 440nm.

1.2 Preparation of solution

Precisely weigh 9.06 mg of the hematocritin high phosphate reference substance and dissolve it in a 10 ml volumetric flask to prepare a methanol reference substance stock solution with a concentration of 0.906 mg/ml.

Preparation of the test solution: Accurately pipette 0.1 mL of the sample solution, add 1 mL of methanol, and filter with a 0.22 μm microporous filter membrane after ultrasonic treatment.

1.3 Exclusiveness inspection

As can be seen from Figure 1, hematocritin perchlorate can be detected under the chromatographic conditions under item 1.1 with good specificity. Figure 1 Diagram of the hematocritin perchlorate standard.

1.4 Drawing of standard curve

Precisely pipette a certain amount of hematocritin perchlorate reference substance stock solution and accurately prepare it with methanol to obtain hematocritin perchlorate concentration of 0.00906mg·mL-1, 0.01812mg·mL-1, and 0.03624mg·mL. -1, 0.07248mg·mL-1, 0.14496g·mL-1 samples, detected according to chromatographic conditions. Use the peak area of the component to be measured (Y) to perform curve fitting regression on the sample concentration (X). The standard curve of hematosparin perchlorate is Y=2E+07X-33740R2=0.9988, and the linear range is: 0.00906~0.14496 mg/ml. Figure 2 Hedradin perchlorate standard curve.

1.5 Precision inspection

The results of the precision experiment of drug components are shown in Table 1. The RSD value of drug peak area was 1.94%, which was less than 2.0%, indicating that the precision of this method was good.

Table 1 Precision test results

Example 2

Solid lipid nanoparticles were prepared using a high-temperature emulsification-low-temperature solidification method. Weigh 100 mg of glyceryl monostearate, 200 mg of soybean lecithin, and 20 mg of hematocritin perchlorate and dissolve them in 2 ml of ethanol. Dissolve with ultrasonic to form an organic phase. Keep it at 80°C for later use. Another 120 mg of poloxamer 407 (F127) was dissolved in 4 ml of deionized water, and heated in a water bath at 80°C to dissolve it as the water phase. With stirring at 80°C and 1000r/min, slowly drip the organic phase into the water phase, continue stirring at 80°C for 30 minutes to form colostrum, pour into 20mL (0~2°C) ice water, continue stirring for 1 hour, and cool to room temperature. That is, SLN suspension is obtained. (Add 50 mg sodium dodecyl sulfate to the water phase to prepare negatively charged solid lipid nanoparticles).

Preparation of hematocritin perchlorate solid lipid nanoparticles freeze-dried powder: add 9% glucose solution of the same volume as the solid lipid nanoparticles into a vial, shake gently, and pre-freeze at -80°C for 12 hours , and then freeze-dried at -80°C for 12 hours to obtain SLN freeze-dried powder, which was sealed and dried for storage.

Example 3 Investigation of Poloxamer Dosage

Examine the effect of the added amount of F127 on the stability and formability of SLN. Keep other components unchanged. Weigh 30 mg, 60 mg, 120 mg, 180 mg, and 240 mg of F127 respectively. Prepare blank SLN according to the method of Example 2, and measure its stability. and particle size, the results are shown in Table 4.

Table 4 Effect of different dosages of poloxamer on preparations

Embodiment 4 Investigation on the feeding ratio of glyceryl monostearate and soybean lecithin

The effect of the feed ratio of monostearate to soybean lecithin on the preparation of SLN was investigated. The other components were kept unchanged, and the feed ratios of monostearate to soybean lecithin were 2:1, 1:1, 1:2, 1:3, and 1:4, respectively. Blank SLN was prepared according to Example 2, and its stability and particle size were measured. The results are shown in Table 5.

Table 5 Effect of the feeding ratio of glyceryl monostearate and soybean lecithin on the preparation

Example 5 Water Phase Volume Investigation

To examine the effect of aqueous phase volume on the preparation and stability of SLN, F127 was dissolved in 2ml, 4ml, 6ml, 8ml, and 10ml deionized water respectively, keeping other components unchanged, and preparing SLN according to Example 2, and measuring its stability. properties and particle size, the results are shown in Table 6.

Table 6 Effect of water phase volume on preparation

Example 6 Investigation of Stirring Speed

SLN was prepared according to the method of Example 2, and the effect on the stability of the preparation at different stirring speeds of 500r/min, 1000r/min, and 1500r/min was investigated. The stabilization time of the preparation at 1000r/min and 1500r/min is longer than that at 500r/min. However, the rotational speed at 1500r/min is too high, prone to liquid splashing, sample loss, and high energy consumption, so the rotational speed of 1000r/min is selected.

Table 7 Effect of stirring rate on preparation process

Example 7 Star Point Effect Design Optimization SLN

6.1 Taking the feeding ratio of glyceryl monostearate to soybean lecithin (A), the dosage of poloxamer 408 (B), and the volume of the aqueous phase (C) as factors, determine the investigation level of each factor based on the single factor results, and use the particle size as an indicator. The factor level design table is shown in Table 2, and the experimental design and results are shown in Table 3.

Table 2 Design experimental factors and levels

Table 3BBD experimental design and results

6.2 Best process verification experiments

According to the best formula obtained through BBD optimization (100 mg of glyceryl monostearate, 200 mg of soybean lecithin, and 120 mg of poloxamer in 4 ml of water), prepare 3 copies in parallel, and measure the particle size and dispersion.

6.3 Determination of drug loading capacity

Weigh 10 mg, 15 mg, 20 mg and 25 mg of hematocritin perchlorate standard respectively and dissolve them in the oil phase to prepare perchlorate-SLN and measure the encapsulation efficiency and drug loading capacity.

6.4BBD Design Optimization SLN

It can be seen from Table 3 that the primary and secondary factors affecting SLN particle size are A>C>B respectively. Use Design-Expert.V8.0.6 software to perform equation fitting on the data in Table 3. The results are shown in Table 8. ^The equation is Y=+7029.9588-6016.59835A-34.88524B-880.88321C+1.95274AB+303.35821AC+1.32292BC+3325.23947A 2 +00.10602B ² +48.41875C ² R ² =0.92 44. Figure 3 Star point effect and contour lines picture.

Table 8 Variance analysis of quadratic polynomial regression model

6.5 Determination of encapsulation efficiency by low-speed centrifugation

Accurately measure 2ml of SLN into a centrifuge tube, centrifuge at 4000r/min for 10min, accurately transfer the supernatant and dilute it 10 times with methanol to break the emulsification, pass through a 0.22μm filter membrane, and measure the hematocrit content by liquid phase detection, recorded as W1, this is The content of the drug encapsulated in the lipid nanoparticles; in addition, an equal amount of SLN was accurately pipetted, diluted with methanol and demulsified by ultrasonic, and the drug content was measured and recorded as W2 respectively. This is the encapsulated and The total amount of unencapsulated drug. Specific encapsulation rate calculation formula: Encapsulation rate = W1/W2 × 100%

The calculation formula for drug loading is: drug loading = W1/W _total × 100% W _total : total mass of SLN

The results are shown in Table 9. When 25 mg of hematocritin perchlorate standard was added, the encapsulation rate was 81.97%. Therefore, it was finally determined to add 20 mg of the drug and the drug loading amount was 0.072%.

Table 9 Determination of encapsulation efficiency and drug loading capacity

6.6 Verification test

According to the optimal recipe optimized by BBD, three portions were prepared in parallel and their particle sizes were measured. The average value was 599 and RSD was 8.72. The particle size diagram is shown in Figure 4. Figure 4 SLN particle size diagram.

Validation example: Pharmacodynamic evaluation of hematocritin perchlorate solid lipid nanoformulation

The hedradin perchlorate solid lipid nanoparticle preparation prepared in Example 2 of the present invention improves the pain threshold of neuropathic rats by reducing the expression of NMDAR 1, and has a therapeutic effect on injurious neuralgia.

1 Experimental Materials

1.1 Animals

SD rats, male, weight range 200-220 g, certificate number SCXK (Beijing) 2016-0002, purchased from Beijing Sibeifu Experimental Animal Technology Co., Ltd., and kept in the animal room of Beijing University of Chinese Medicine.

1.2 Reagents and drugs

Hematin perchlorate; Hematin perchlorate solid lipid nanoparticles; Absolute ethanol; Normal saline; Chloral hydrate; 10% neutral formalin fixative; 4% paraformaldehyde; penicillin ;NMDAR1 antibody;Fenbid phenol coffee tablets.

1.3 Instruments

OLYMPUSBX51 microscope (Beijing Ruichi Hengye Instrument Technology Co., Ltd.); FA1204B analytical balance (Shanghai Precision Instrument Co., Ltd.); electric thermostatic water bath (Tianjin Taist Instrument Co., Ltd.), etc.

2Experimental methods

2.1 Establishment of CCI animal model

The CCI model was constructed according to the method of Bennett et al. ^[5-6] : anesthesia was administered intraperitoneally with 10% chloral hydrate (0.3ml/100g). The rat was fixed prone on the rat board, depilated and disinfected. The skin was incised about 1cm below the femur of the right hind limb parallel to the femur. The subcutaneous tissue and muscles were bluntly separated to expose the sciatic nerve. About 7mm was freed and made of No. 4.0 chromium. Make 4 loose ligations on the catgut. The distance between ligations is about 1mm. Pay attention to the tightness during ligation. The muscle will twitch slightly when tying. Use 0.9% sodium chloride injection to flush the incision. After ligation, proceed gradually. The layers were sutured, and 400,000 U of penicillin was injected intramuscularly to prevent infection. After the rats woke up, they were raised in separate cages. The animals in the sham operation group only exposed the sciatic nerve but did not ligate it, and the rest of the operations were the same as described above.

2.2 Animal grouping and administration

Seventy SD rats were randomly divided into model group (Model group), sham operation group (Sham group), hematocritin perchlorate group (1.2mg/200g, DP group), hematocritin perchlorate group Solid lipid nanoparticle group (1.2mg/200g, DP-NM group) and positive drug (Positive group), 12 animals in each group. The sham operation group and the model group were given normal saline by gavage. Administration was started the next day after the modeling was completed, once a day until the end of the experiment.

2.3 Drug efficacy evaluation

2.3.1PWMT measurement

The paw withdrawal mechanical threshold (PWMT) of the rats was measured 1 day before surgery, 7 days, 14 days, and 21 days after surgery. After the rats adapted to the environment, the Von Frey probe was used to vertically stimulate the rats in the order of increasing quality. From the skin in the middle of the sole of the rat's foot to the hind limb of the rat, the rat twitches or lifts its foot to escape, and record the value at this time (unit: g) ^[7] . Measure 5 times continuously with an interval of 10 minutes.

2.3.2PWTL determination

The paw withdrawal thermal latency (PWTL) of the rats was measured 1 day before surgery, 7 days, 14 days, and 21 days after surgery. After the rats were stable, a thermal pain meter was used to measure the response of the skin in the middle of the left hind limb to thermal stimulation. The PWTL threshold is the time from irradiation to the paw withdrawal reaction of lifting the paw, avoiding or licking the paw ^[8] . The measurement was repeated 5 times at intervals of 10 minutes.

2.3.3 Specimen collection

A section of the sciatic nerve in the ligated area was taken from the injury site on the side of the rat model, and conventional paraffin-embedded sections were prepared, HE-stained sections were mounted, and light microscopy was performed.

2.3.4 Determination of NMDAR1 in spinal dorsal root ganglia by immunohistochemistry

After measuring the pain threshold on day 21, the rat was deeply anesthetized, its limbs were fixed, the abdominal cavity was opened under the xiphoid process, the transverse septum was passed through, the heart was exposed, a perfusion needle was inserted into the left ventricle and fixed, the right atrial appendage was cut, and the heart was perfusion. First, quickly perfuse with 4°C sterile saline until the outflowing liquid becomes clear, and then perfuse quickly and then slowly with 4% paraformaldehyde to fix it for about 30 minutes. Then find the sciatic nerve at the center of the right hind limb femur, and go up the sciatic nerve to find the L4-6 spinal ganglion connected to it, separate and remove it, fix it in 10% neutral formaldehyde for 48 hours, embed it in paraffin, and slice it into 4mm serial sections ^[7] . Staining was performed according to the instructions of the immunohistochemistry kit, and the analysis results were observed using the image-pro plus 6.0 image analysis system. The results were expressed as average optical density.

3Experimental results

3.1PWMT measurement results

Compared with the sham operation group, the rats in the CCI model group showed significant mechanical pain from day 1 to day 21; compared with the model group, the PWMT of the Positive group, DP group and DP-NM group increased significantly (p< 0.01), there was also an increase in the DP-NM group (p<0.05), indicating that a certain dose of hematocritin perchlorate can effectively inhibit mechanical pain in the CCI model; the DP-NM group and the DP-L group had the same dosage, but For the analgesic effect of mechanical pain, the DP-NM group was better than the DP group (p<0.05 at 14 days), indicating that the solid lipid nanoparticle drug delivery system can help improve the anti-mechanical pain effect of hematosine perchlorate. See Table 1 for specific results.

Table 1 Comparison of PWMT in rats of each group at different time periods (g, `x ± s, n = 6)

Table 1Comparison of PWMT in different time periods of rats in each group

Note: Compared with the model group, ^* p＜0.05, ^** p＜0.01; compared with the DP group, ^# p＜0.05

3.2 Results of thermal stimulation paw withdrawal latency measurement

Compared with the sham operation group, rats in the CCI model group showed significant thermal allergy from day 1 to day 21. Compared with the model group, the PWTL in the Positive group and the DP-NM group increased significantly (p < 0.01), and the DP-M group also increased (p < 0.05), indicating that a certain dose of dracoside perchlorate can effectively inhibit mechanical pain in the CCI model.

Table 2 Comparison of PWTL of rats in each group at different time periods (s, `x±s, n=6)

Table 2Comparison of PWTL in different time periods of rats in each group

Note: Compared with the model group, ^* p<0.05, ^** p<0.01.

3.3 Pathological sections of rat sciatic nerve tissue

The structure of the interstitium and nerve fibers around the sciatic nerve of animals in the Sham group was intact and neatly arranged, with no abnormal changes. Compared with the Sham group, animals in the Model group had vacuoles in the cytoplasm, irregular nuclei, and many mast cells. Compared with the Sham group, the degree of sciatic nerve edema and vacuolar degeneration in the administration group was reduced. The specific results are shown in Figure 5. Figure 5: Effect of tincture on pathological changes of rat sciatic nerve (HE staining, 200×).

3.4 Immunohistochemistry results

There were significant differences between the DP-H group, DP-M group and Model group (p<0.05). This shows that an appropriate amount of hemodrin perchlorate can reduce the pain sensitivity of sciatica rats and inhibit the expression of NMDAR 1 in the spinal cord.

Table 3 Expression of NMDAR 1 in spinal cord of rats in each group (n=6,`x±s)

Table 3The expression of NMDAR-1 in spinal cord of rats in each group

Note: Compared with the model group, ^* p<0.05

4.1 Liquid phase conditions of hematocrit sample

Chromatographic column: ACQUITY UPLCTM BEH C18 (1.7μm, 2.1×50mm); mobile phase: acetonitrile-water; gradient elution: 0min, 20% acetonitrile; 6min, 35% acetonitrile; 6.1min, 20% acetonitrile; column temperature 35°C , sample temperature 4℃;

Detection wavelength: 440nm.

4.2 Preparation release method

Precisely transfer 3 mL of each portion into 100 mL of 0.5% SDS release external solution. Add 10 mL of release medium to the pretreated dialysis bag and immerse it in the release external solution. Shake in a constant temperature water bath at 37°C. The two groups were respectively at 0.167h, 0.5h, 1h, 2h, 3h, 4h, 6h, 8h, 10h, 12h, 1d, 2d, 3d, 6d, 9d, 12d, 15d, 18d, 21d, 24d, 27d, 30d. Draw 2 mL of release medium from the dialysis bag (and add an equal amount of release medium at the same temperature), measure the content of hematocrit by HPLC, calculate the cumulative release percentage of the drug, and fit the release kinetic model.

The cumulative release percentage Qn at different time points was calculated according to the following formula.

Qn is the cumulative release percentage at the nth point (%); V is the release medium volume (mL); Cn is the drug mass concentration measured at the nth point (mg·mL-1); 2 is sampling 2mL; W is administration Total amount (mg).

4.3 Experimental Results

Taking the cumulative release percentage at each time point as the ordinate and the sampling time as the abscissa, draw the in vitro drug release curves of the hematocrit original drug and the hematocrit emulsion. The results are shown in Figure 6. Figure 6 shows the in vitro drug release curves of the hematocritin original drug and the hematocritin emulsion. In vitro drug release profile of hematocritin emulsion. It can be seen from Figure 6 that the cumulative release rate of the hematocrit emulsion is higher than that of the hematogenin original drug group.

The above are only the preferred embodiments of the present invention. It should be pointed out that those of ordinary skill in the art can also make several improvements and modifications without departing from the principles of the present invention. These improvements and modifications can also be made. should be regarded as the protection scope of the present invention.

Claims (10)

1. A method for preparing dracoside perchlorate solid lipid nanoparticles, comprising: A) Mix glyceryl monostearate, soybean lecithin and hematocritin perchlorate, and dissolve them with a solvent to obtain an organic phase; B) an aqueous solution of poloxamer as the aqueous phase; C) Add the organic phase to the water phase and cool to obtain. 2. The preparation method according to claim 1, characterized in that the mass ratio of the glyceryl monostearate and soybean lecithin is 1: (1-3); The mass ratio of the glyceryl monostearate, soybean lecithin and hematocritin perchlorate is (8-12): (18-22): (1-3). 3. The preparation method according to claim 1, characterized in that the concentration of the aqueous solution of poloxamer is 10 to 40 mg/mL. 4. The preparation method according to claim 1, characterized in that the mass ratio of the glyceryl monostearate, soybean lecithin and poloxamer is (8~12): (18~22): (10~ 14). 5. The preparation method according to claim 1, characterized in that the solvent in step A) is ethanol; the dissolution is ultrasonic dissolution, and the power of the ultrasonic is 200~300HZ; the temperature of the dissolution is 70~300HZ. 80℃; Step B) the poloxamer aqueous solution is obtained by dissolving poloxamer in water; the dissolution temperature is 70-80°C. 6. The preparation method according to claim 1, characterized in that step C) The specific steps of adding the organic phase into the water phase are as follows: under stirring conditions at 70-80°C, the organic phase is slowly dripped into the water, and stirred at 80°C for 20-30 minutes; the stirring speed is 1000-1400 r/min; The cooling is specifically: cooling in ice water at 0 to 2°C, stirring for 50 to 70 minutes, and then continuing to cool at 25 to 30°C. 7. A hematocritin perchlorate solid lipid nanoparticle, characterized in that it is prepared by the preparation method described in any one of claims 1 to 6. 8. A dracotin perchlorate drug, characterized in that it comprises the dracotin perchlorate solid lipid nanoparticles according to claim 7. 9. The medicine according to claim 8, wherein the dosage form of the medicine is one or more of powder, suspension, tablet, capsule, granule or powder. 10. Application of the hedradin perchlorate solid lipid nanoparticles according to claim 7 in the preparation of medicines for the treatment of injurious neuralgia.