Double-layer micro-capsule granule product for soothing nerves and helping sleep and preparation method of double-layer micro-capsule granule product

Abstract

The invention discloses a double-layer microcapsule granule product for soothing nerves and helping sleep and a preparation method thereof, and relates to the technical field of traditional Chinese medicines. The double-layer micro-capsule granule product comprises the following raw materials: a double-layer micro-capsule coating layer and a double-layer micro-capsule granule, the double-layer micro-capsule coating layer comprises the following components: sodium alginate-chitosan and ethyl cellulose; the double-layer micro-capsule granules comprise the following raw materials in parts by weight: 150-200 parts of litsea coreana, 250-300 parts of gastrodia elata and 100-150 parts of American ginseng. According to the physicochemical characteristics of different traditional Chinese medicine effective components, corresponding extraction modes are selected in a targeted manner, and the stability of the medicine is improved; the extracted effective components are wrapped by double-layer wrapping materials, so that the sensory feeling of the traditional Chinese medicine is improved, and the palatability of the medicine and the taking compliance of a patient are improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, and particularly relates to a double-layer micro-capsule granule product for soothing the nerves and helping sleep and a preparation method thereof. BACKGROUND

[0002] In recent years, with the continuous increase of social life rhythm and work and study pressure, more and more people have sleep disorders to some extent. The main clinical manifestations are insomnia, dreaminess, difficulty falling asleep, light sleep, easy wake, short sleep time, circadian rhythm disorder and the like, and are accompanied by depression, anxiety, tension and the like. Insufficient sleep can cause memory function decline, attention function decline, and even affect the normal physiological activities of the cardiovascular system, the digestive system and the motor system, and cause the disorder of immune function and endocrine function. Epidemiological studies show that the population with sleep disorder related symptoms accounts for about 12.1% in the world. Sleep disorders not only affect people's normal life, but also further affect people's physical and mental health.

[0003] At present, the intervention and treatment of sleep disorders in modern medicine mainly include sedative and soothing drugs, and the use methods include oral use, aroma therapy, acupoint massage, acupoint sticking, external use of gel and the like. However, these methods are often slow in effect, long in treatment course, and improper use may cause other unnecessary damage. Therefore, it is necessary to find a treatment method with higher safety, faster effect and more convenient use method.

[0004] Gastrodia elata, Panax quinquefolius and Ilex hainanensis have the effects of suppressing liver yang, tonifying qi and nourishing yin, soothing the nerves and helping sleep. The effective components are extracted by different extraction technologies, and the effective components are wrapped by a coating layer to form a double-layer micro-capsule. The adaptability of oral taking of traditional Chinese medicine is improved, and the sensory perception is improved. In view of the defects of the current sleep aid products, a double-layer micro-capsule granule for soothing the nerves and helping sleep is invented. The effective components are extracted by a scientific extraction method, and the effective components of traditional Chinese medicine are treated by a double-layer coating. A double-layer micro-capsule granule is developed, the stability, palatability and convenience of taking of the medicine are significantly improved, and the compliance of patients is improved. SUMMARY

[0005] The present application aims at solving the defects in the prior art, and provides a double-layer micro-capsule granule product for soothing the nerves and helping sleep and a preparation method thereof.

[0006] In order to achieve the above-mentioned purpose, the present application adopts the following technical scheme: A double-layer micro-capsule granule product for soothing the nerves and helping sleep and a preparation method thereof, the raw materials of the double-layer micro-capsule granule product include a double-layer micro-capsule coating and a double-layer micro-capsule granule. The components of the double-layer microcapsule coating include: sodium alginate-chitosan, ethyl cellulose; The raw materials of the double-layer microcapsule granules include: 150-200 parts of Ilex asprella, 250-300 parts of Gastrodia elata, and 100-150 parts of Panax quinquefolium.

[0007] Preferably, the mass ratio of the raw materials in the double-layer microcapsule granules is Gastrodia elata: Panax quinquefolium: Ilex asprella = 5-7: 2-5: 3-6.

[0008] A preparation method of a double-layer microcapsule granule product for soothing the nerves and helping sleep, characterized in that it comprises the following steps: S1: preparing the raw materials in proportion; S2: extracting the effective components of Gastrodia elata, Panax quinquefolium, and Ilex asprella to obtain an extract; S3: reducing and concentrating the effective component extract under pressure, and freeze-drying to obtain a powder; S4: forming a sustained-release coating layer by ethyl cellulose to prepare microspheres; S5: preparing a quick-release coating material; S6: packaging the product.

[0009] Preferably, in S2, the effective component extraction method for Gastrodia elata is: Gastrodin is extracted by low-temperature enzymatic hydrolysis. After slicing, Gastrodia elata is pretreated with cellulase and pectinase at 30-50°C and pH 4.0-6.0 to destroy the cell wall and improve the dissolution rate of gastrodin.

[0010] Preferably, in S2, the effective component extraction method for Panax quinquefolium is: Panax notoginseng is extracted by supercritical CO2, and 60%-80% ethanol is used as a carrier agent to selectively extract ginsenosides Rb1 and Rg1 at 35-55°C and 20-30 MPa to avoid the destruction of heat-sensitive components by high temperature.

[0011] Preferably, in S2, the effective component extraction method for Ilex asprella is: The polyphenols and flavonoids in Ilex asprella are extracted by 30-50 kHz ultrasonic waves combined with 60%-80% ethanol.

[0012] Preferably, in S4, the sustained-release coating layer is formed by ethyl cellulose to prepare microspheres, and the specific preparation method is: The sustained-release coating liquid is prepared according to the prescription amount, the dried powder of the effective components of Panax quinquefolium prepared by extraction is placed in a fluidized bed, the parameters of the fluidized bed are adjusted, and after the preheating temperature reaches the high set value of the material temperature, the coating is started. After the coating is completed, drying is continued to obtain microspheres.

[0013] Preferably, in S5, the preparation method of the quick-release coating material is: The extracted Gastrodia elata and Ilex asprella effective component freeze-dried powder and the prepared microspheres are mixed with 3% to 12% sodium alginate and 2% to 8% glycerol to obtain a mixed solution, the mixed solution is prepared into microcapsules by a microcapsule instrument, is dripped into CaCl2 at a constant speed, is magnetically stirred until completely hardened, is rinsed with 0.85% to 0.9% physiological saline and then is placed in a 0.8% to 1% chitosan solution for stirring to form a film, sodium alginate / chitosan microcapsules are prepared, and finally the microcapsules are completely rinsed with 0.85% to 0.9% physiological saline, are frozen at -80°C for 24 hours, and are quickly placed in a vacuum freeze dryer for treatment and then taken out.

[0014] Preferably, the product is packaged in a single-dose independent package.

[0015] Preferably, the product is used for soothing the nerves, relieving depression, improving sleep quality, prolonging sleep time and preventing insomnia.

[0016] The present application has the following beneficial effects: 1. According to the physical and chemical properties of different traditional Chinese medicine effective components, the present application selects corresponding extraction methods, thereby improving the stability of the medicine.

[0017] 2. The effective components extracted by the present application are wrapped with double-layer coating materials, thereby improving the sensory experience of traditional Chinese medicine, and improving the palatability of the medicine and the compliance of patients.

[0018] 3. Unlike traditional decoctions, the double-layer microcapsule granules can be directly taken orally or brewed for taking, so that patients can select the taking mode according to the actual situation.

[0019] 4. The product of the present application is finally packaged independently, thereby improving the storage time of the medicine, facilitating transportation and having a long shelf life. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 FIG. 5 is a diagram for significant difference analysis of 5-HT content in the present application; Figure 2 FIG. 6 is a diagram for significant difference analysis of TNF-α content in the present application; Figure 3 FIG. 7 is a diagram for significant difference analysis of GABA content in the present application; Figure 4 FIG. 8 is a diagram for bacterial flora difference at the genus level in the present application; Figure 5 FIG. 9 is a diagram for bacterial flora difference at the door level in the present application; Figure 6 FIG. 10 is a diagram for HE staining of intestinal tissue in the present application. DETAILED DESCRIPTION

[0021] The technical solutions of the present application will be further described in detail in combination with specific embodiments.

[0022] Embodiment 1: A kind of tranquilizing sleep-aiding double-layer micro-capsule granule product, which is made of the following raw materials and their mass fractions: old eagle tea 150-200 parts, Gastrodia elata 250-300 parts, American ginseng 100-150 parts, sodium alginate-cystose, ethyl cellulose.

[0023] The preparation method of the tranquilizing sleep-aiding double-layer micro-capsule granule includes the following steps: S1. Gastrodia elata is subjected to low-temperature enzymatic extraction of gastrodin. After slicing, the Gastrodia elata is pretreated with cellulase and pectinase at 40℃ and pH 5.0 to destroy the cell wall and improve the dissolution rate of gastrodin.

[0024] S2. American ginseng is extracted by supercritical CO2. Ginsenosides Rb1 and Rg1 are selectively extracted at 45℃ and 30MPa with 70% ethanol as the entraining agent to avoid the destruction of heat-sensitive components by high temperature.

[0025] S3. 40kHz ultrasonic waves are used in combination with 60% ethanol to extract polyphenols and flavonoids from old eagle tea.

[0026] S4. The effective ingredient extract is reduced pressure concentrated and dried, and the powder is obtained after freeze-drying.

[0027] S5. The coating liquid (ethyl cellulose: polyethylene glycol 4000: 95% ethanol = 10:1:50) is prepared according to the prescription amount. The dried powder of the effective ingredient extracted from American ginseng is placed in a fluidized bed. The fluidized bed parameters are adjusted. After the preheating temperature reaches the high set value of the material temperature, the coating is started. After the coating is completed, the drying is continued.

[0028] S6. The freeze-dried powder of the effective ingredient extracted from Gastrodia elata and old eagle tea and the microspheres prepared in claim 8 are mixed with 4% sodium alginate and 6% glycerol. The mixture is prepared into microcapsules by a microcapsule instrument. The microcapsules are dropped into 0.2mol / L CaCl2 at a uniform speed, magnetically stirred for 10min until they are completely hardened. After being washed with 0.85% physiological saline, they are placed in a 0.8% chitosan solution and stirred for 10min to form a film. Sodium alginate / chitosan microcapsules are prepared. Finally, they are completely washed with 0.85% physiological saline, frozen at -80℃ for 24h, and then quickly placed in a vacuum freeze-drying machine for 72h (conditions: cold trap temperature -70℃, vacuum degree 1000mbar).

[0029] Embodiment 2: Study on the effect of the product prepared in embodiment 1 on sleep disorders Typical case: Example 1 comprehensive treatment effect of the product on sleep disorder mice Randomly divide 48 SPF BALB / c 6-8 weeks male mice into 6 groups, respectively: blank control group (intraperitoneal injection of normal saline, intraperitoneal injection amount: 0.1 mL / 10g), sleep disorder model group (intraperitoneal injection of PCPA to induce sleep disorder mouse model, intraperitoneal injection amount: 0.1 mL / 10g), experimental group: eagle tea group, American ginseng group, Gastrodia elata group, eagle tea+American ginseng+Gastrodia elata group. Gavage administration, the administration period is 14 days. 30 min after the last administration, the eyeball is taken to collect blood, and the brain tissue of the mouse is taken. The brain homogenate is determined by ELISA for the content of 5-HT, GABA and TNF-α in the brain. The results are shown in Tables 1-3, and the significance analysis is as follows Figures 1-3 .

[0030] Table 1 5-HT content (pg / mL) Blank group Model group Eagle tea group American ginseng group Gastrodia elata group Eagle tea + American ginseng + Gastrodia elata group 207.75±0.33 111.18±2.79 153.05±7.48 152.63±1.58 181.58±0.74 228.56±2.59 Table 2 TNF-α content determination (ng / L) Blank group Model group Eagle tea group American ginseng group Gastrodia elata group Eagle tea + American ginseng + Gastrodia elata group 417.02±5.90 934.81±1.06 782.43±56.77 744.07±10.03 610.88±1.59 466.23±30.45 Table 3 GABA content determination (mol / L) Blank group Model group Eagle tea group American ginseng group Gastrodia elata group Eagle tea + American ginseng + Gastrodia elata group 4.28±0.04 2.74±0.03 3.35±0.23 3.55±0.07 3.73±0.02 4.75±0.02 From the above experimental results, it can be seen that the 5-HT and TNF-α contents of the eagle tea+American ginseng+Gastrodia elata group are reduced, indicating that the reduction of 5-HT content caused by PCPA is reversed, and the effect of the eagle tea+American ginseng+Gastrodia elata group is the most obvious; the TNF-α content is reduced, indicating that the inflammatory pathway is blocked; the GABA content is increased, and the inhibitory neurotransmitter content is increased, indicating that the neural activity is reduced. And each administration group has a significant difference (p<0.0001) with the model group. The sustained-release microcapsule of the application has the effects of soothing the nerves and helping sleep, and has a certain therapeutic effect on sleep disorders.

[0031] Example 3: Study of the effect of the product of example 1 on sleep disorders Typical case: Effect of example 1 product on intestinal flora structure of sleep disorder mice Randomly divide 48 SPF BALB / c 6-8 weeks male mice into 6 groups, respectively: blank control group (intraperitoneal injection of normal saline, intraperitoneal injection amount: 0.1 mL / 10g), sleep disorder model group (intraperitoneal injection of PCPA to induce sleep disorder mouse model, intraperitoneal injection amount: 0.1 mL / 10g), experimental group: eagle tea group, American ginseng group, Gastrodia elata group, eagle tea+American ginseng+Gastrodia elata group. Gavage administration, the administration period is 14 days. Two days before the last administration, the feces of the mice are collected, and 16s rRNA sequencing is performed to analyze the intestinal flora of the mice, as shown in the following figure.

[0032] Comparison of intestinal flora abundance at genus level. As can be seen from the figure, the number of bacteria with large abundance difference mainly includes Bacteroides, Glucomastix, Lactobacillus and Prevotella. Bacteroides exists in the human intestine and has a symbiotic relationship with humans, helps to decompose food and produce nutrients and energy needed by the body, and belongs to beneficial bacteria. Compared with the blank group, the abundance of Bacteroides in the hawk tea + American ginseng + Gastrodia elata group was improved. Glucomastix may cause animal infection, and compared with the blank group, the abundance of Glucomastix in the hawk tea + American ginseng + Gastrodia elata group was reduced. Prevotella can help decompose protein and carbohydrate food, and compared with the blank group and other administration type groups, the abundance of Prevotella in the hawk tea + American ginseng + Gastrodia elata group was increased. Different administration types can cause changes in the abundance of different genera, and in general, the abundance of beneficial bacteria is increased and the abundance of harmful bacteria is reduced.

[0033] Comparison of intestinal flora abundance at door level. As can be seen from the figure, the number of bacteria with large abundance difference mainly includes Bacteroidetes, Firmicutes, Actinobacteria and Ossipovia. The abundance of Bacteroidetes is increased, and the abundance of the rest of the bacterial door is reduced.

[0034] In summary, according to the experimental results, the hawk tea + American ginseng + Gastrodia elata group can change the composition of the intestinal flora of mice, increase the abundance of beneficial bacteria, reduce the abundance of harmful bacteria, promote the composition of the intestinal flora, improve the intestinal environment, and thus affect the health status of mice.

[0035] Example 4: Study on the effect of the product of example 1 on sleep disorders Typical case: Effect of example 1 product on intestinal structure of sleep disorder mice Forty-eight 6-8-week-old male SPF BALB / c mice were randomly divided into six groups: blank control group (intraperitoneal injection of normal saline, intraperitoneal injection amount: 0.1 mL / 10g), sleep disorder model group (intraperitoneal injection of PCPA to induce sleep disorder mouse model, intraperitoneal injection amount: 0.1 mL / 10g), experimental group: hawk tea group, American ginseng group, Gastrodia elata group, hawk tea + American ginseng + Gastrodia elata group. Gavage administration, the administration period is 14 days. Thirty minutes after the last administration, the colon tissue of the mice was taken. The colon tissue was sectioned, HE stained, and the changes in intestinal tissue structure were observed. The results are as follows Figure 6 . Among them: A: blank control group: the intestinal morphology is complete, the intestinal wall microstructure is basically normal, the intermuscular nerve plexus nerve cells (green arrow) are clearly visible, the crypt structure in the mucosa is normal, and the goblet cells are rich. The epithelial cells are arranged in order.

[0036] B: Model group: The intestinal morphology was complete, the intermuscular nerve plexus nerve cells (green arrow) were clearly visible, the crypt structure in the mucosa was normal, the goblet cells were rich, and the epithelial cells were arranged in order; a large number of lymphocytes were visible (black arrow) in the local mucosa, and the mucosal muscle layer was broken through.

[0037] C: Eagle tea group: The intestinal morphology was complete, the intermuscular nerve plexus nerve cells (green arrow) were clearly visible, the crypt structure in the mucosa was normal, the goblet cells were rich, and the epithelial cells were arranged in order; a few individual epithelial cells were visible (orange arrow), a large number of lymphocytes were visible (black arrow) in the local mucosa, and the mucosal muscle layer was broken through.

[0038] D: American ginseng group: Part of the intermuscular nerve plexus nerve cells was pyknosis (green arrow); the mucosal part of the intestinal gland structure was damaged, the crypt was damaged and degenerated with inflammatory cell infiltration (red arrow), and part of the epithelial cells were pyknosis (orange arrow); a large number of lymphocytes were visible (black arrow) in the local mucosa, and the mucosal muscle layer was broken through.

[0039] E: Gastrodia elata group: The intestinal morphology was complete, the intestinal wall thickness was relatively thin, the intermuscular nerve plexus nerve cells (green arrow) were clearly visible, the crypt structure in the mucosa was normal, and the goblet cells were rich; a large number of lymphocytes were visible (black arrow) in the local mucosa, and the mucosal muscle layer was broken through.

[0040] F: Eagle tea + American ginseng + Gastrodia elata group: The intestinal morphology was complete, the crypt structure in the mucosa was normal, and the goblet cells were rich; a large number of lymphocytes were visible (black arrow) in the local mucosa, and the mucosal muscle layer was broken through.

[0041] From the above results, it can be seen that the eagle tea + American ginseng + Gastrodia elata group can prevent the deterioration of the intestinal structure caused by sleep disorders in mice, and has a protective effect on the intestine.

[0042] Example 5: Study on the release performance detection of the product in Example 1 The product in Example 1 was placed in 20 mL of PBS, and a dialysis membrane was added, and placed in a 37°C shaking bed at a speed of 75 r·min~1. 1 mL of PBS liquid was taken out at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h, respectively. After taking out 1 mL of liquid each time, 1 mL of the same temperature medium 1mL PBS was immediately supplemented to ensure that the total volume of the liquid was 20 mL. The sample was filtered through a 0.22 μm filter membrane and detected by HPLC.

[0043] Chromatographic conditions: Hypersil C18 (4.6 mm x 250 mm, 5 µm); mobile phase A was acetonitrile, mobile phase B was 0.1% phosphoric acid water, gradient elution was used (0-7 min, 6% A; 7-8 min, 6%-10% A; 8-10 min, 10%-18% A; 10-15 min, 18% A; 15-40 min, 18%-21% A; 40-42 min, 21%-26% A; 42-46 min, 26%-32% A; 46-66 min, 32%-33.8% A; 66-71 min, 33.8%-38% A; 71-77 min, 38%-49% A; 77-78 min, 49%-50% A; 78-82 min, 50% A; 82-83 min, 50%-51% A; 83-88 min, 51%-60% A; 88-90 min, 60%-65% A; 90-97 min, 65% A; 97-114 min, 65%-85% A; 114-117 min, 85%-6% A; 117-140 min, 6% A); flow rate 1.0 mL / min; column temperature 35°C; detection wavelength 200 nm, injection volume 10 µL.

[0044] After HPLC detection, the release performance was summarized to obtain Table 4 below.

[0045] Table 4 Release performance of granules

[0046] The above results show that the release performance of the product of Example 1 of the present application is good, and can achieve the state of rapid falling asleep and long-term stable sleep.

[0047] The above describes only the preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, any person skilled in the art can make equivalent replacement or change according to the technical solution and inventive concept of the present application within the technical range disclosed by the present application, which should be covered in the protection scope of the present application.

Claims

1. A double-layered microcapsule granule product for calming the nerves and aiding sleep, characterized in that, The raw materials included in the bilayer microcapsule granule product are a bilayer microcapsule coating and bilayer microcapsule granules. The components of the bilayer microcapsule encapsulation include: sodium alginate, chitosan, and ethyl cellulose; The raw materials for the double-layer microcapsule granules include: 150-200 parts of eagle tea, 250-300 parts of gastrodia elata, and 100-150 parts of American ginseng.

2. The double-layer microcapsule granule product for calming the nerves and aiding sleep, and its preparation method, as described in claim 1, is characterized in that... The mass ratio of raw materials in the double-layer microcapsule granules is Gastrodia elata: American ginseng: eagle tea = 5~7: 2~5: 3~6.

3. A method for preparing the sedative and sleep-aiding double-layer microcapsule granule product according to claim 1 or 2, characterized in that, Includes the following steps: S1: Prepare the raw materials according to the proportions; S2: Extract the effective components from Gastrodia elata, American ginseng, and eagle tea to obtain an extract; S3: The active ingredient extract is concentrated and dried under reduced pressure, and then freeze-dried to obtain a powder; S4: Microspheres were prepared by forming a sustained-release coating using ethyl cellulose; S5: Preparation of immediate-release coating material; S6: Product packaging.

4. The method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 3, characterized in that, In S2, the extraction method of the effective components of Gastrodia elata is as follows: Gastrodin was extracted from Gastrodia elata by low-temperature enzymatic hydrolysis. After slicing, Gastrodia elata was pretreated with cellulase and pectinase at 30-50℃ and pH 4.0-6.0 to disrupt the cell wall and improve the dissolution rate of gastrodin.

5. The method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 3, characterized in that, In S2, the extraction method of the effective components of American ginseng is as follows: Supercritical CO2 extraction was used to extract American ginseng, with 60%–80% ethanol as an entrainer. Ginsenosides Rb1 and Rg1 were selectively extracted at 35–55℃ and 20–30 MPa to avoid damaging their heat-sensitive components at high temperatures.

6. The method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 3, characterized in that, In S2, the extraction method of the effective components of Eagle Tea is as follows: Polyphenols and flavonoids of Lao Ying tea were extracted using 30-50kHz ultrasound combined with 60%-80% ethanol.

7. The method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 3, characterized in that, In step S4, a sustained-release coating is formed using ethyl cellulose to prepare microspheres. The specific preparation method is as follows: Prepare the sustained-release coating solution according to the prescription, place the dried powder of the active ingredients of American ginseng obtained by extraction into a fluidized bed, adjust the fluidized bed parameters, and start coating after the preheating temperature reaches the set value of the material temperature. After coating is completed, continue drying to obtain microspheres.

8. The method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 7, characterized in that, In step S5, the method for preparing the immediate-release coating material is as follows: The freeze-dried powders of the effective components of Gastrodia elata and Eagle Tea, along with the prepared microspheres, were mixed with 3%–12% sodium alginate and 2%–8% glycerol to obtain a mixture. The mixture was then used to prepare microcapsules using a microencapsulation apparatus. The microcapsules were added dropwise into CaCl2 at a uniform rate and magnetically stirred until completely hardened. After rinsing with 0.85%–0.9% physiological saline, the microcapsules were placed in a 0.8%–1% chitosan solution and stirred to form a film, thus obtaining sodium alginate / chitosan microcapsules. Finally, the microcapsules were completely rinsed with 0.85%–0.9% physiological saline, frozen at ~80℃ for 24 hours, and then quickly placed in a vacuum freeze dryer for further processing before being removed.

9. A method for preparing a double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 3, characterized in that, The product is packaged in single-dose, individually packaged form.

10. A double-layer microcapsule granule product for calming the nerves and aiding sleep according to claim 1 or 2, characterized in that, The product is used to calm the nerves, improve sleep quality, prolong sleep time, and prevent insomnia.

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