NL2033485B1

NL2033485B1 - Rhizoma pinelliae-rhizoma atractylodis macrocephalae-rhizoma gastrodiae composite granules, as well as preparation method and application thereof

Info

Publication number: NL2033485B1
Application number: NL2033485A
Authority: NL
Inventors: Sun Yunting; Li Xinyue; Xu Nan; Ye Keke; Yang Feifei; Wang Shuling
Original assignee: Univ Hangzhou Normal
Priority date: 2022-04-22
Filing date: 2022-11-08
Publication date: 2023-11-07
Also published as: CN114748599A

Abstract

Disclosed are Rhizoma pinelliae—Rhizoma atractylodis macrocephalae—Rhizoma gastrodiae composite granules as well as a preparation method and an application thereof. The composition is prepared from. an aqueous extract of plants and pharmaceutical excipients by granulation using a high—speed stirring granulator, wherein the aqueous extract of plants is prepared from prepared Rhizoma pinelliae, Rhizoma gastrodiae, Poria, Exocarpium citri rubrum, Rhizoma atractylodis macrocephalae, Radix et rhizoma glycyrrhizae, Rhizoma zingiberis recens and Eîuctus jujubae. The Rhizoma pinelliae—Rhizoma atractylodis macrocephalae—Rhizoma gastrodiae granules not only retain the effects of the original prescription, such as drying dampness and resolving phlegm, and calming liver wind, but also overcome the disadvantages that a decoction is time—consuming and energy—consuming for temporary preparation, and, the decoction. is easy to go moldy and, go bad after long—term storage; the granules are smaller in dosage and more convenient in administration, carrying, storage and transport.

Description

RHISOMA PINELLIAE-RHIZOMA ATRACTYLODIS MACROCEPHALAE-RHIZOMA

GASTRODIAE COMPOSITE GRANULES, AS WELL AS PREPARATION METHOD AND

APPLICATION THEREOF

TECHNICAL FIELD

The present application belongs to the technical field of pharmaceutical preparations, and particularly relates to Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules, as well as a preparation method and an appli- cation thereof.

BACKGROUND ART

Hypertension is a major public health problem around the world. It is estimated that 1.39 billion people worldwide had been suffering from hypertension as of 2010. In China, the prevalence of hypertension is high, but the treatment management rate is low.

In the past decades, the incidence of hypertension in China has increased significantly, especially in rural areas. Obesity, age and living habits are the common risk factors of hypertension. Hy- pertension is also a major and independent risk factor of cardio- vascular diseases. Clinical trials have proved that lowering blood pressure can reduce cardiovascular diseases and premature death.

Many medicinal preparations have been approved for the treatment of hypertension, but many of them have potential side effects or are only used for individual patients. Moreover, hypertension has many complications and is difficult to treat. Hypertension may cause heart failure, stroke, vision problems, kidney disease and other problems.

A Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-

Rhizoma gastrodiae composition has the effects of drying dampness and resolving phlegm, and calming liver wind. The composition is mainly used for treating the syndromes of dizziness and headache induced by wind-phlegm invading upward, chest tightness and nau- sea, whitish glossy coating of the tongue, and slippery pulse.

This prescription, as a commonly used prescription for treating wind-phlegm syndrome, is still widely applied in the clinical practice of traditional Chinese medicine. For example, the pre- scription is used to treat hypertensive phlegm-dampness congestion type and phlegm-dampness type Meniere's syndrome, epilepsy caused by phlegm confusing the heart and orifices, meridian stroke and sequelae caused by wind-phlegm stagnation. The wind-phlegm syn- dromes are originated from phlegm produced by spleen dampness, phlegm stagnation and clearing of yang, as well as the liver wind agitation, and wind-phlegm invading upward. The predecessors said that “no phlegm, no dizziness” and “all wind syndromes such con- vulsion and dizziness attribute to the liver”. Due to the liver wind agitation, a person may feel objects are shaking due to diz- ziness; in addition to the wet phlegm invading upward and the tur- bid yin invading upward, the dizziness becomes even worse, and the person feels that both the sky and earth are spinning and then vomits and revolts. Resolving phlegm and calming wind are appro- priate for treatment. In the prescription, Rhizoma pinelliae is hot and warm in nature, and good for drying dampness and resolving phlegm, and calming adverse-rising energy to control nausea to treat the root causes; and Rhizoma gastrodiae is sweet in taste and neutral in nature, and good for calming liver wind and reliev- ing dizziness to treat the symptoms. The two medicinal materials are combined to treat both the root causes and the symptoms, and both are monarch medicines specially for resolving phlegm and calming wind. Rhizoma atractylodis macrocephalae is used as a min- ister medicine for strengthening the spleen and drying dampness, and has good effects on clearing dampness, resolving phlegm and relieving dizziness in compatibility with Rhizoma pinelliae and

Rhizoma gastrodiae. Poria as an adjuvant has the functions of strengthening the spleen and excreting dampness, and especially can treat the root cause for phlegm in compatibility with Rhizoma atractylodis macrocephalae; Exocarpium citri rubrum can regulate gi to resolve phlegm, so that gi becomes smooth to resolve phlegm.

Radix et rhizoma glycyrrhizae is used as a guide to regulate and neutralize the medicinal materials, and the medicinal materials are combined to jointly play a role in resolving phlegm and calm- ing wind. Wind is calmed to resolve phlegm, and dizziness will be self-cured.

The Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-

Rhizoma gastrodiae granules have the advantages of rapid absorp- tion, fast onset of action and powerful effect, and also overcome the disadvantages that a decoction is time-consuming and energy- consuming for temporary preparation, and the decoction is easy to go moldy and go bad after long-term storage; the granules are smaller in dosage and more convenient in administration, carrying, storage and transport. The Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules reduce the labor inten- sity to a greater extent, and make up the market gap; moreover, the economic benefit is considerable and the advantage is obvious.

SUMMARY

One objective of the present invention is to provide Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules in order to solve the problems existing in the prior art, which provides a theoretical support for the subsequent mass production of the Rhizoma pinelliae-Rhizoma atractylodis mac- rocephalae-Rhizoma gastrodiae granules.

The present invention is realized by the following technical solution: the Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-

Rhizoma gastrodiae composite granules include an agueous extract of plants and pharmaceutical excipients, wherein the aqueous ex- tract of plants is an aqueous extract of the following ingredients based on parts by weight: 5-10 parts of prepared Rhizoma pinelli- ae, 5-10 parts of Rhizoma gastrodiae, 5-10 parts of Poria, 5-15 parts of Exocarpium citri rubrum, 10-20 parts of Rhizoma at- ractylodis macrocephalae, 1-5 parts of Radix et rhizoma glycyr- rhizae, 1-5 parts of Rhizoma zingiberis recens and 1-5 parts of

Fructus jujubae; the pharmaceutical excipients include a filler, a wetting agent and a binding agent; wherein the filler is one or more of sucrose, starch, dextrin, lactose, mannitol and sodium alginate; the wetting agent is 80%-95% ethanol (volume content) or water; wherein the filler accounts for 30%-60% of the weight of the Rhi-

zoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastro- diae composite granules; the wetting agent accounts for 5%-15% of the weight of the Rhizoma pinelliae-Rhizoma atractylodis macro- cephalae-Rhizoma gastrodiae composite granules; and the binding agent accounts for 0%-5% of the weight of the Rhizoma pinelliae-

Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules; preferably, the binding agent is one of povidone, polyeth- ylene glycol and hydroxy propyl cellulose; preferably, the specific dosages of the ingredients in the aqueous extract of plants are as follows based on parts by weight: 10 parts of prepared Rhizoma pinelliae, 5 parts of Rhizoma gastro- diae, 5 parts of Poria, 5 parts of Exocarpium citri rubrum, 10 parts of Rhizoma atractylodis macrocephalae, 3 parts of Radix et rhizoma glycyrrhizae, 2 parts of Rhizoma zingiberis recens and 1 part of Fructus jujubae.

Another objective of the present invention is to provide a preparation method for the Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules, including the following steps: (1) weighing prepared Rhizoma pinelliae, Rhizoma gastrodiae,

Poria, Exocarpium citri rubrum, Rhizoma atractylodis macrocepha- lae, Radix et rhizoma glycyrrhizae, Rhizoma zingiberis recens and

Fructus jujubae according to the proportions, respectively, pul- verizing separately and then mixing these medicinal materials to obtain a mixture of medicinal materials; (2) adding 1.0-2.0% enzyme reagent I (m/m) to the mixture of medicinal materials prepared in step (1), and pretreating the mix- ture for 12-24 h; (3) adding water of 10, 8 and 8 times in mass to the mixture of medicinal materials pretreated in step (2) to adjust a pH value to 1-6, 6-9 and 6-9, respectively, adding 0.1-1.0%enzyme reagent I (m/m) and 0.1-1.0% enzyme reagent II (m/m) to the first extraction and the third extraction, respectively, performing extraction at 37-60°C three times, 0.5-1 h for each extraction, filtering the extract, merging the filtrate, and concentrating the filtrate to

1/3-2/3 of the original amount to obtain an aqueous extract con- centrate; (4) adding alcohol to the aqueous extract concentrate until an alcohol concentration reaches 60-80% (v/v), placing the mixture 5 in a refrigerator at 4°C for 24 h, evenly mixing and centrifugating the mixture, collecting the supernatant, concentrating the super- natant to 1/3-2/3 of the original amount, and performing drying under reduced pressure and pulverization to obtain dry paste pow- der; (5) opening a high-speed stirring granulator, setting parame- ters, adding the dry paste powder and a filler into a mixing tank, starting a stirring blade for dry mixing, uniformly adding a bind- ing agent and a wetting agent during wet mixing, starting a cut- ter, discharging granules after granulation is completed, and dry- ing the granules at a constant temperature of 70°C for 30-60 min; and (6) packaging; wherein the enzyme reagent I is one or more of cellulase, hemicellulase and pectinase, the enzyme reagent I is one or more of pepsin, trypsin and papain, and the enzyme reagent I is one or more of pepsin, trypsin and papain; preferably, the parameters of the high-speed stirring granu- lator in step (4) are as follows: a dry mixing time is 3-5 min, a wet mixing time is 5-15 min, a granulating time is 1-4 min, a stirring speed for dry mixing is 400-600 rpm, a stirring speed for wet mixing is 400-600 rpm, a stirring speed for granulation is 1,000-1,500 rpm, and a cutting speed is 2,000-3,000 rpm.

The third objective of the present invention is to provide an application of the Rhizoma pinelliae-Rhizoma atractylodis macro- cephalae-Rhizoma gastrodiae composite granules in preparing a drug for treating or improving phlegm-dampness stagnation type

Meniere’s syndrome.

The fourth objective of the present invention is to provide a drug for treating or improving phlegm-dampness stagnation type

Meniere's syndrome, including Rhizoma pinelliae-Rhizoma at- ractylodis macrocephalae-Rhizoma gastrodiae composite granules.

The present invention has the advantages that:

The Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-

Rhizoma gastrodiae granules prepared by the preparation method of the present invention have been proved to be safe and effective by quality inspection and pharmacodynamic experiment. In the present invention, based on the full consideration of practicability and effectiveness as well as previous researches, the Rhizoma pinelli- ae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules not only retain the effects of the original prescription, such as drying dampness and resolving phlegm, and calming liver wind, but also overcome the disadvantages that a decoction is time-consuming and energy-consuming for temporary preparation, and the decoction is easy to go moldy and go bad after long-term storage; the Rhi- zoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastro- diae granules are smaller in dosage and more convenient in admin- istration, carrying, storage and transport. The Rhizoma pinelliae-

Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules re- duce the labor intensity to a greater extent, shorten the labor time, greatly lessen people’s burden, and make up the market gap; the economic benefit is considerable and the advantage is obvious.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a finished product of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules.

DETAILED DESCRIPTION OF THE EMBODIMENTS

As mentioned above, in view of the deficiency in the prior art, the inventor proposed the technical solutions in the present invention after long-term study and a plenty of practice. The main basis at least includes: 1) in the prescription, Rhizoma pinelliae is hot and warm in nature, and good for drying dampness and re- solving phlegm, and calming adverse-rising energy to control nau- sea to treat the root causes; and Rhizoma gastrodiae is sweet in taste and neutral in nature, and good for calming liver wind and relieving dizziness to treat the symptoms. The two medicinal mate- rials are combined to treat both the root causes and the symptoms, and both are monarch medicines specially for resolving phlegm and calming wind. Rhizoma atractylodis macrocephalae is used as a min- ister medicine for strengthening the spleen and drying dampness, and has good effects on clearing dampness, resolving phlegm and relieving dizziness in compatibility with Rhizoma pinelliae and

Radix et rhizoma glycyrrhizae is used as a guide to regulate and neutralize the medicinal materials, and the medicinal materials are combined to jointly play a role in resolving phlegm and calm- ing wind. Wind is calmed to resolve phlegm, and dizziness will be self-cured. It is especially effective in preparing a drug for treating or improving phlegm-dampness stagnation type Meniere’s syndrome. 2) The technologies of mixing and granulation are inte- grated by using a high-speed stirring granulator, thereby both saving time and meeting GMP requirements, reducing cross contami- nation, and greatly improving the efficiency. 3) The Rhizoma pi- nelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules of the present invention not only retain the effects of the original prescription, such as drying dampness and resolving phlegm, and calming liver wind, but also overcome the disad- vantages that a decoction is time-consuming and energy-consuming for temporary preparation, and the decoction is easy to go moldy and go bad after long-term storage; and moreover, the Rhizoma pi- nelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules are smaller in dosage and more convenient in administra- tion, carrying, storage and transport.

In order to make the objectives, technical solutions and ad- vantages of the present invention more clearly, the present inven- tion will be further explained below in detail in combination with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are used only for explaining, rather than limiting, the present invention. In addi- tion, the technical features involved in various embodiments of the present invention described below may be combined with each other, as long as there is no conflict.

In the first aspect, a preparation method for Rhizoma pinel- liae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae compo- site granules includes the following steps: (1) weighing 5-10 parts by weight (the same below) of Rhizoma pinelliae, 5-10 parts of Rhizoma gastrodiae, 5-10 parts of Poria, 5-15 parts of Exocarpium citri rubrum, 10-20 parts of Rhizoma at- ractylodis macrocephalae, 1-5 parts of Radix et rhizoma glycyr- rhizae, 1-5 parts of Rhizoma zingiberis recens and 1-5 parts of

Fructus jujubae, respectively, pulverizing separately and then mixing these medicinal materials to obtain a mixture of medicinal materials; (2) adding 1.0-2.0% enzyme reagent I to the mixture of me- dicinal materials prepared in step (1), and pretreating the mix- ture for 12-24 h; wherein the enzyme reagent I is one or more of cellulase, hemicellulase and pectinase; (3) adding water of 10, 8 and 8 times in mass to the mixture of medicinal materials pretreated in step (2) to adjust a pH value to 1-6, 6-9 and 6-9, respectively, adding 0.1-1.0%enzyme reagent I and 0.1-1.0% enzyme reagent II to the first extraction and the third extraction, respectively, performing extraction at 37-60°C three times, 0.5-1 h for each extraction, filtering the extract, merging the filtrate, and concentrating the filtrate to 1/3-2/3 of the original amount to obtain an aqueous extract concentrate; wherein the enzyme reagent II is one or more of pepsin, trypsin and papain, and the enzyme reagent II is one or more of pepsin, trypsin and papain; (4) adding alcohol to the aguecus extract concentrate until an alcohol concentration reaches 60-80%, placing the mixture in a refrigerator at 4°C for 24 h, evenly mixing and centrifugating the mixture, collecting the supernatant, concentrating the supernatant to 1/3-2/3 of the original amount, and performing drying under re- duced pressure and pulverization to obtain dry paste powder; (5) opening a high-speed stirring granulator, setting parame- ters, adding the dry paste powder and a filler into a mixing tank, starting a stirring blade for dry mixing, uniformly adding a bind-

ing agent and a wetting agent during wet mixing, starting a cut- ter, discharging granules after granulation is completed, and dry- ing the granules at a constant temperature of 70°C for 30-60 min; wherein the filler is one or more of sucrose, starch, dextrin, lactose, mannitol and sodium alginate, and the filler accounts for 30%-60% of the weight of the Rhizoma pinelliae-Rhizoma at- ractylodis macrocephalae-Rhizoma gastrodiae composite granules; and the binding agent accounts for 0%-5% of the weight of the Rhi- zoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastro- diae composite granules; and (6) packaging.

Preferably, the wetting agent is 80%-95% ethancl or water.

Preferably, the wetting agent accounts for 5%-15% of the weight of the Rhizoma pinelliae-Rhizoma atractylodis macrocepha- lae-Rhizoma gastrodiae composite granules.

Preferably, the binding agent is one of povidone, polyeth- ylene glycol and hydroxy propyl cellulose.

Preferably, the parameters of the high-speed stirring granu- lator in step (5) are as follows: a dry mixing time is 3-5 min, a wet mixing time is 5-15 min, a granulating time is 1-4 min, a stirring speed for dry mixing is 400-600 rpm, a stirring speed for wet mixing is 400-600 rpm, a stirring speed for granulation is 1,000-1,500 rpm, and a cutting speed is 2,000-3,000 rpm.

In the second aspect, Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules are prepared by the above-mentioned method, including an aqueous extract of plants and pharmaceutical excipients, wherein the aqueous extract of plants is an aqusous extract of the following ingredients based on parts by weight: 5-10 parts of prepared Rhizoma pinelliae, 5-10 parts of Rhizoma gastrodiae, 5-10 parts of Poria, 5-15 parts of

Exocarpium citri rubrum, 10-20 parts of Rhizoma atractylodis mac- rocephalae, 1-5 parts of Radix et rhizoma glycyrrhizae, 1-5 parts of Rhizoma zingiberis recens and 1-5 parts of Fructus jujubae; and the pharmaceutical excipients include a filler, a wetting agent and a binding agent.

In the third aspect, an application of the Rhizoma pinelliae-

Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules in preparing a drug for treating or improving phlegm- dampness stagnation type Meniere’s syndrome is provided.

The technical solutions of the present invention will be fur- ther explained in combination with some preferred embodiments, but the experimental conditions and setting parameters in the embodi- ments should not be considered as a limitation to the basic tech- nical solutions of the present invention. Besides, the protection scope of the present invention is not limited to the following em- bodiments.

Example 1 10 g of prepared Rhizoma pinelliae, 5 g of Rhizoma gastrodi- ae, 5 g of Peoria, 5 g of Exocarpium citri rubrum, 10 g of Rhizoma atractylodis macrocephalae, 3 g of Radix et rhizoma glycyrrhizae, 2 g of Rhizoma zingiberis recens and 1 g of Fructus jujubae were weighed respectively, pulverized separately and then mixed to ob- tain a mixture of medicinal materials. 1.5% sacroiliac enzyme rea- gent I (cellulase) was added to the mixture of medicinal materi- als, and the mixture was pretreated for 24 h. Water of 10, 8 and 8 times in mass was added to adjust a pH value to 5, 7 and 8, re- spectively, 1.0% enzyme reagent I (pepsin) and 0.5% enzyme reagent

I (trypsin) were added to the first extraction and the third ex- traction, respectively, extraction was performed at 50°C three times, 1 h for each extraction, the extract was filtered, the fil- trate was merged and concentrated to 1/3 of the original amount to obtain an aqueous extract concentrate. Alcohol was added to the aqueous extract concentrate until an alcohol concentration reached 60%, the mixture was placed in a refrigerator at 4°C for 24 h and then evenly mixed and centrifugated, the supernatant was collected and concentrated to 2/5 of the original amount, and the concen- trate was dried under reduced pressure and pulverized to obtain dry paste powder. A high-speed stirring granulator was started, parameters (a dry mixing time was 4 min, a wet mixing time was 8 min, a granulating time was 4 min, a stirring speed for dry mixing was 400 rpm, a stirring speed for wet mixing was 500 rpm, a stir- ring speed for granulation was 1,500 rpm, and a cutting speed was 2,000 rpm) were set, 2 kg of dry paste powder and 1 kg of filler

(starch : mannitol = 3:2) were added into a mixing tank, a stir- ring blade was started for dry mixing, 0.3 kg of povidone ethanol solution (a mass ratio of povidone to 90% ethanol was 5:95) was uniformly added during wet mixing, a cutter was started, granules were discharged after granulation was completed, and dried at a constant temperature of 70°C for 60 min.

According to the appearance and the granularity, moisture and solubility test items of granules in the Chinese Pharmacopoeia (2020 Edition), the Rhizoma pinelliae-Rhizoma atractylodis macro- cephalae-Rhizoma gastrodiae composite granules prepared in example 1 were tested for the indicators. The results are as shown in Ta- ble 1, and the picture of finished products is as shown in FIG. 1.

Table 1 Test Results oe een

Granularity (95.21+ 1.41%

Example 2

The extraction method (SBEE) used in the present invention, alcohol extraction (AE) method and water extraction (WE) method were optimized, respectively, and extracting solutions were pre- pared by the three methods. The gastrodin (HPLC method), glycyr- rhetinic acid (HPLC method), extract with a molecular weight of S 1,000 Da (constant weight method), HPLC integral total area, total polysaccharide (UV spectrophotometry), and dry extract yield (con- stant weight method) were taken as indicators for comprehensive evaluation, the SBEE solution, the WE solution and the AE solution were compared to preliminarily determine a better extraction meth- od. The test results are as shown in Table 2- Table 7.

Table 2 Test Results of Gastrodin Content

Gastrodin Content (mgeg )

Solution ee ee Lc LL a

Table 3 Test Results of Glycyrrhetinic Acid Content

Solution

Table 4 Test Results of Extract with a Molecular Weight of < 1,000 Da

Solution 1,000 Da (geg’}

Table 5 Test Results of HPLC Integral Total Area

HPLC Total Area ¥ ac RSD (%)

Solution

Table € Test Results of Total Polysaccharide

Extracting Total Polysaccharide {Absorbancy)

Solution

SBEE 0.276 0.281 0.279 027900025 | 090 5 Table 7 Test Results of Dry Extract Yield

Solution

The data of various indicators were standardized according to the following formulas TD) and 2), and the standardized X', ; values were weighted and then summed to obtain the Y values of the com- prehensive evaluation indicators (the results are as shown in Ta- ble 8).

EE XS D

Fo ome EERE where X, ; is a content of a component j in a sample solution i, Xs is a mean value of the component j in the sample solution i, 34 is a standard deviation of the component j, and X';, is a standardized value.

Table 8 Contents of Various Indicator Components and Results of Standardization Processing

Test Gastrodin | Glycyr- Extract with | HPLC Total | Total Polysac- Dry Ex- Y

Solution (mgeg’) rhetinic a Molecular | Area charide (Ab- tract Val

Acid Weight of s sorbancy) gg’) ue (ugg?) 1,000 Da (gg)

SBEE 1.0643 50.6280 0.1536 14,474.6 0.279 0.4930 9.79

EEE EE Er

WE 0.9510 (- | 26.5298 0.0757 {- 13,513.0{- | 0.220 0.4182 (- - 0.3928) | (-0.1793) 0.9476) 0.5244) (-0.0899) 0.1307) 4.29 11

AE 0.8135 16.3314 0.1017 {- 13,181.2(- | 0.123 0.3339 (- - (1.0666) | (-0.8635) 0.1611) 1.0965) {-1.3810) 1.3445) 8.70 14

Notes: (1) The “g” in the unit of measurement for each compo- nent is equivalent to the mass of a TCM in the prescription; 2) The data in the brackets are standardized values; 3) Y = (gastrodin + glycyrrhetinic acid + extract with a mo- lecular weight of < 1,000 Da} /3x7+{polysaccharide + HPLC total peak area}/2x2+ dry extract x1.

The values of the comprehensive evaluation was listed in a descending sequence: Ysssz>Ygz>Yas, preliminarily indicating that the

SBEE method was optimal.

Example 3

SD rats were selected, comnon feed and a certain amount of kaolin feed were put into a cage at the same time, and the rats were adaptively fed for two weeks. At Week 3, the rats were ran- domly divided into five groups, 8 rats in each group, namely a normal control group, a blank control group, a high dose group, a medium dose group and a low dose group, respectively. The rats in the normal control group and the blank control group were perfused with plain boiled water once, and the rats in the administration groups were administrated intragastrically with an aqueous solu- tion of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-

Rhizoma gastrodiae granules at 2 g/kg (low dose group), 4 g/kg (medium dose group) and 8 g/kg (high dose group), respectively.

After 0.5 h, except the normal control group, the rats in the oth- er four groups were centrifugated at 250 rpm in a low-speed cen- trifuge for 5 min and then taken out. The above operations were repeated after 2 min until the time for the rats in the centrifuge added up to a total of 20 min. Then the kaolin intake amount of the rats in the five groups within 3 days was observed. Average values were calculated as the experimental results. An inter-group significance test of average values and an inter-group comparison (t test) were performed for the rats. The results are as shown in

Table 9. The average value of the kaolin intake amounts of the rats was processed to obtain R;' as an evaluation value. 50 mice were selected (including 25 males and 25 females) and divided into five groups, namely a normal control group, a blank control group, a high dose group, a medium dose group and a low dose group, respectively. The mice in the normal control group and the blank control group were perfused with plain boiled water, and the mice in the other groups were administrated intragastrically with an aqueous solution of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules at 2 g/kg (low dose group), 4 g/kg (medium dose group) and 8 g/kg (high dose group), respectively. After 0.5 h, except the normal control group, the mice in the other four groups were centrifugated at 250 rpm in a low-speed centrifuge for 5 min, and then immediately placed on a 60 cm-long horizontal bar with a diameter of 0.8 cm, and the shortest time from standing steadily on the bar at the end of cen- trifugation to being capable of smoothly walking on the bar was recorded for each mouse. Average values were calculated as the ex- perimental results. An inter-group comparison (t test) was per- formed. After the experiment, the mice trembled with erecting hair and made counterclockwise circular movements. The mice could not stand stably on the 60 cm-long horizontal bar with a diameter of 0.8 cm, and fell down from the bar freely; afterwards, the mice could hold the bar, the body of each mouse was pendulous without falling down, the hair erected, and the mice still trembled; and finally, the mice could stand steadily and walk freely on the bar without hair erecting and trembling symptom. An inter-group sig- nificance test of average values and an inter-group comparison (t test) were performed for the results. The results of the shortest time for the mice in each group to stand steadily on the rod after the rotation experiment are as shown in Table 10. The average val- ue of the shortest time for the mice to stand steadily on the rod was processed to obtain R,' as an evaluation value.

Ry’ and R;’ were processed according to the relational expres- sion: Rg=(R;'+R;*)/2. The Rs value was obtained as an indicator for evaluating the influence on the rats and mice with phlegm-dampness stagnation type Meniere’s syndrome, whichever is greater. The re- sults are as shown in Table 11.

Table 9 Comparison of Kaolin Intake Amount of Rats in the

Groups after the Rotation Experiment (qg) (#54) "Animal No.

Gow 12 3 4 5s 6 7 8 LR

HighDose 2.18 256 0.87 115 212 108 0.98 122 15240.6569% 191

Group

Medium Dose 1.47 1.78 2.01 2.43 1.02 2.73 1.13 2.52 1.89+0.647"*" 1.54

Group

Low Dose 264 27 2.24 2.12 3.04 2.31 2.56 207 2.46+0.333°* 1.18

Group

Normal Con- 0.05 001 0.12 0.07 0.13 0.09 0.08 0.02 0.07+0.0434 trol Group

Blank Control 3,32 341 2.06 2.37 3.19 2.98 3.51 2.44 2.91+0.547

Group

Notes: D Compared with the blank control group, 4P<0.001, ’P <0.01, 'F»0.05; compared with the normal control group,

p< 0.001.

GRE, Wywas the average kaolin intake amount of the rats in the blank control group, and x was the kaolin intake amount of the rats in each group.

Table 10 Shortest Time for the Mice in Each Group to Stand

Steadily on the Rod after the Rotation Experiment (min, zE)

CAnimalNo.

Gow 2346

HighDoseGroup 10 9 9 8 12 15 8 11 1025:2.385% 346

Medium Dose Group 12 13 16 15 12 11 17 14 13.75+2.12V% 2.58

Low Dose Group 21 17 19 15 16 18 17 18 17.63+1.85" # 2.01

Normal Control 04 05 0 0.7 0.3 0.9 0.2 0.5 0.44+0.28À

Group

Blank Control Group + 37 + 40 + + 35 30 35.50+4.20

Notes: (1) “+” represents that the animal died. @ Compared with the blank control group, 4P< 0.001, 'P<0.01; compared with the normal control group, fp<0.001. 3) Ral, “gas the average shortest time for the mice in the blank control group to stand steadily on the rod, and * was the shortest time for the mice in each group to stand steadily on the rod.

Table 11 Comprehensive Evaluation Ry; Values for the Animals in Each Group After the Rotation Experiment mlm fw fw

Note: Rg = (R{"+R;")/2

The results of the rotation stimulating experiment of the rats showed that as compared with the blank control group, the ka- olin intake amount of the rats in the other groups were obviously decreased, wherein the decrease in the kaolin intake amount was obvious in the high dose group and the medium dose group but not obvious in the low dose group, and there was no significant dif-

ference (P>0.05), indicating that high-dose and medium-dose Rhi- zoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastro- diae granules had stronger effect against the rotation stimulated dizziness of rats.

The results of the rotation stimulating experiment of the mice showed that as compared with the blank control group, the shortest time for the mice in all the other groups to stand stead- ily on the bar was significantly decreased. The sequence of the R,’ value was high dose group > medium dose group > low dose group, indicating that the Rhizoma pinelliae-Rhizoma atractylodis macro- cephalae-Rhizoma gastrodiae granules had stronger effect against the rotation stimulated dizziness of mice.

In the rotation stimulating experiment of the mice, the re- sults in the experimental groups were obviously different from those in the normal control group, and part of the animals in the blank control group died, indicating that the method of rotation modeling for inducing dizziness to animals had too strong physical stimulation, and the time and speed of the rotation need to be further optimized; but no animal died in the other experimental groups, indicating that the Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae granules had stronger effect against the rotation stimulated dizziness of mice.

Finally, it should be noted that the above-mentioned embodi- ments are only the preferred embodiments of the present invention, and not intended to limit the present invention. Although the pre- sent invention is explained in detail by reference to the above- mentioned embodiments, modification to the technical solutions recorded in the above-mentioned embodiments, or equivalent substi- tution to a part of the technical features may be made by those skilled in the art. Any modification, equivalent substitution or improvement within the spirit and principle of the present inven- tion should fall into the protection scope of the present inven- tion.

Claims

CONCLUSIONS

1. Preparation method for Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules, comprising the following steps: (1) weighing out a weight of 5 to 10 parts by weight (same below) of Rhizoma pinelliae, 5 to 10 parts Rhizoma gastrodiae, 5 to 10 parts Poria, 5 to 15 parts Exocarpium citri rubrum, 10 to 20 parts Rhizoma atractylodis macrocephalae, 1 to 5 parts Radix et rhizoma glycyrrhizae, 1 to 5 parts Rhizoma zingiberis recens and 1 to 5 parts Fructus jujubae, it separately pulverizing and then mixing these medicinal materials to obtain a mixture of medicinal materials; (2) adding 1.0 to 2.03 enzyme reagent I to the mixture of medicinal materials prepared in step (1), and pretreating the mixture for 12 to 24 hours; wherein the enzyme reagent I is one or more of cellulase, hemicellulase and pectinase; {3) adding water with a mass of 10, 8 and 8 times to the mixture of medicinal materials pretreated in step (2) to adjust the pH value to 1 to 6, 6 to 9 and 6 respectively to 9, adding 0.1 to 1.0% enzyme reagent II and 0.1 to 1.0% enzyme reagent III to the first extraction and third extraction, respectively, performing extraction three times at 37 to 60 °C, 0 .5 to 1 hour before each extraction, filtering the extract, combining the filtrate and concentrating the filtrate to 1/3 to 2/3 of the original amount to obtain an aqueous extract concentrate; wherein the enzyme reagent II is one or more of pepsin, trypsin and papain, and the enzyme reagent III is one or more of pepsin, trypsin and papain; (4) adding alcohol to the aqueous extract concentrate until an alcohol concentration reaches 60 to 80%, placing the mixture in a refrigerator at 4°C for 24 hours, mixing evenly and centrifuging the mixture, collecting the supernatant, concentrating the supernatant to 1/3 to 2/3 of the original amounts, and performing dry-

mixing under reduced pressure, and pulverizing to obtain dry pasta powder; (5) opening a high-speed stirring granulator, setting parameters, adding the dry paste powder and a filler into a mixing tank, starting a dry mixing stirring blade, adding a binder and a wetting agent evenly during wet mixing, starting a cutter, discharging granules after granulation is completed, and drying the granules at a constant temperature of 70°C for 30 to 60 minutes; wherein the filler is one or more of sucrose, starch, dextrin, lactose, mannitol and sodium alginate, and the filler constitutes 30 to 60% of the weight of the composite granule of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae; and the binder represents 0 to 5% by weight of the composite granules of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae; and (6) packaging.

Preparation method for Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules according to claim 1, wherein the humectant is 80% to 95% alcohol or water.

3. Preparation method for Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules according to claim 1 or 2, wherein the wetting agent constitutes 5% to 15% of the weight of the Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules.

Preparation method for composite granules of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae according to claim 1, wherein the binder is one of povidone, polyethylene glycol and hydroxypropyl cellulose.

5. Preparation method for composite granules of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae vol-

according to claim 1, wherein the parameters of the high speed stirring granulator in step (5) are as follows: a dry mixing time is 3 to 5 min, a wet mixing time is 5 to 15 min, a granulating time is 1 to 4 min, a stirring speed for dry mixing is 400 to 600 rpm, a wet mixing stirring speed is 400 to 600 rpm, a granulating stirring speed is 1,000 to 1,500 rpm, and a cutting speed is 2,000 to 3,000 rpm.

Preparation method for composite granules of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae according to claim 1, wherein the specific dosages of the ingredients in the aqueous extract of plants are as follows, based on parts by weight: 10 parts prepared Rhizoma pinelliae, 5 parts Rhizoma gastrodiae, 5 parts Peoria, 5 parts Exocarpium Citri rubrum, 10 parts Rhizoma atractylodis macrocephalae, 3 parts Radix et rhizoma glycyrrhizae, 2 parts Rhizoma zingiberis recens and 1 part Fructus jujubae.

7. Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules prepared according to the method according to any one of claims 1 to 6, wherein the composite granules comprise an aqueous extract of plants and pharmaceutical excipients, wherein the aqueous extract of plants is an aqueous extract of the following ingredients on a parts by weight basis: 5 to 10 parts prepared Rhizoma pinelliae, 5 to 10 parts Rhizoma gastrodiae, 5 to 10 parts Poria, 5 to 15 parts Exocarpium citri rubrum, 10 to 20 parts Rhizoma atractylodis macrocephalae, 1 to 5 parts Radix et rhizoma glycyrrhizae, 1 to 5 parts Rhizoma zingiberis recens and 1 to 5 parts Fructus jujubae; and the medicinal excipients include a filler, a wetting agent and a binder.

8. Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules according to claim 7, wherein the aqueous extract of plants is an aqueous extract of the following ingredients on a parts by weight basis: 10 parts prepared Rhizoma pinelliae, 5 parts Rhizoma gastrodiae , 5 parts Poria, 5 parts

Exocarpium citri rubrum, 10 parts Rhizoma atractylodis macrocephalae, 3 parts Radix et rhizoma glycyrrhizae, 2 parts Rhizoma zingiberis recens and 1 part Fructus jujubae.

9. Use of the Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae composite granules according to claim 7 or 8 in the preparation of a medicament for treating or improving Meniere's mucous fluid stagnation syndrome.

A medicament for treating or improving Meniere's mucous fluid stagnation syndrome, wherein the medicament comprises the composite granule of Rhizoma pinelliae-Rhizoma atractylodis macrocephalae-Rhizoma gastrodiae according to claim 7 or 8.