WO2013155994A1

WO2013155994A1 - Red yeast and evening primrose pharmaceutical composition for regulating blood lipids and preparation method therefor

Info

Publication number: WO2013155994A1
Application number: PCT/CN2013/074532
Authority: WO
Inventors: 段震文; 郭树仁; 刘曦
Original assignee: 北京北大维信生物科技有限公司
Priority date: 2012-04-20
Filing date: 2013-04-22
Publication date: 2013-10-24
Also published as: CN103372035A

Abstract

The present invention aims to disclose a pharmaceutical composition for regulating blood lipids (in particular, for treating hypertriglyceridemia) and a preparation method therefor. The present invention also aims to disclose the medicinal uses of the pharmaceutical composition. The red yeast and the evening primrose in the present pharmaceutical composition complement one other and generate an amplification effect, providing the effects of de-humidification, phlegm removal, blood circulation, blood stasis removal, spleen strengthening, and digestion assistance. The present pharmaceutical composition reduces the total cholesterol in blood, increases the high-density lipoprotein concentration, lowers the low-density lipoprotein concentration, and in particular, has substantial effects of lowering triglyceride and treating hypertriglyceridemia. The present invention has a logical manufacturing process that enhances the concentration of the effective component, lovastatin, in the red yeast, while maintaining the active ingredients of different medicine in the prescription, thereby fully using all medicines. The preparation of the present invention uses the concentration of lovastatin as the quality control indicator, and the preparation quality is stable and controllable.

Description

Red yeast evening primrose drug combination for regulating blood lipid and preparation method thereof

The invention relates to a pharmaceutical composition and a preparation method thereof, in particular to a pharmaceutical composition prepared by using a traditional Chinese medicine compound as a raw material and a preparation method thereof, and belongs to the field of traditional Chinese medicine. Background technique

The modern lifestyle has led to an increase in the intake of high-protein, high-cholesterol, and high-sugar foods in people's foods, leading to a marked increase in the incidence of hyperlipidemia. Hyperlipidemia is a pre-existing atherosclerosis and has a significant correlation with the incidence of cerebrovascular disease. Hyperlipidemia is also an important factor in inducing coronary heart disease, arteriosclerosis, fatty liver, diabetes, and obesity. Therefore, prevention and treatment of hyperlipidemia plays an important role in the prevention of cardiovascular and cerebrovascular diseases. It is very necessary to research and develop safe and effective blood lipid regulating drugs and health foods.

Hyperlipidemia refers to excessively high total cholesterol (TC) and/or triglyceride (TG) or high density lipoprotein cholesterol (HDL-C) in the blood. Clinically divided into: hypercholesterolemia (increased serum TC), hypertriglyceridemia (increased serum TG), mixed hyperlipidemia (increased TC, TG), low high density lipoprotein Blood (lower serum HDL-C levels).

The serum TC level in our population is significantly lower than that in the high-risk population of western coronary heart disease. Through a sample survey of 49,252 permanent residents of 18 provinces, autonomous regions and municipalities directly under the Central Government in China: The prevalence of dyslipidemia in 18-year-old residents in China is 18.6%. The prevalence of hypercholesterolemia was 2.9 %; the marginal rate of cholesterol was 3.9 %; the prevalence of hypertriglyceridemia was 11.9%; The rate is 7.4%. The abnormal types of blood lipid metabolism in our country are mainly high triglyceride and low-density lipoproteinemia, which is different from the high-cholesterolemia in western population. It is necessary to find a hypolipidemic drug for the abnormal type of lipid metabolism in Chinese people.

Summary of the invention

SUMMARY OF THE INVENTION It is an object of the present invention to provide a pharmaceutical composition for regulating blood lipids (especially for treating hypertriglyceridemia) and a process for the preparation thereof; and an object of the present invention is to provide a pharmaceutical use of the pharmaceutical composition.

The object of the present invention is achieved by the following technical solutions:

The composition of the drug substance of the pharmaceutical composition of the present invention is:

Red yeast 1 to 5 parts by weight evening primrose 1 to 5 parts by weight;

The drug substance composition of the pharmaceutical composition of the present invention is preferably:

Red yeast 2 parts by weight evening primrose 4 parts by weight;

Red koji 4 parts by weight of evening primrose 2 parts by weight; Red koji 1 part by weight of evening primrose 4 parts by weight;

Red yeast 4 parts by weight evening primrose 1 part by weight;

The pharmaceutical composition of the present invention can be formulated into a clinically acceptable oral preparation according to a conventional method by adding a conventional excipient, including but not limited to a pill, a powder, a tablet, a granule, a capsule, or an oral liquid preparation. The pharmaceutical preparation contains no more than 10 mg of lovastatin in the total amount of the preparation per day.

Preferably, the red yeast in the drug substance of the above pharmaceutical composition may be ordinary red yeast or functional red yeast rice.

Ordinary red yeast rice is red yeast rice (also known as red yeast rice, red yeast rice, red rice, and glutinous rice) which is inoculated on rice by Monascus anka Nakazawa et Sato. It can be purchased from the market or it can be prepared according to the method of GB 4926-2008.

The functional red yeast rice may be commercially available or may be prepared by the following method: (The preparation method is the content of claim 1 of Chinese Patent No. 97116744. 3, 97103970. 4) (1), preparing a culture solution: Glycerin (or wort or potato juice), a mixture of sugar and yeast (or beef extract or peptone) and water, of which glycerin (or wort or potato juice) accounts for 2 to 7 %, sugar accounts for 2 to 6 %, peptone 0~3 %, yeast powder accounts for 0~3 %, beef cream 0~3 %, a small amount of antifoaming agent into soybean oil or peanut oil 2~4%. 0。 The rest of the water, with acetic acid to adjust the pH value of 3. 0~5. 0. (2), per 100 g of glutinous rice, add 30~80 ml of culture solution, high temperature steam (12 灭菌 sterilized. After the temperature is cooled to below 40 ° ,, inoculate the strain of Monascus. The strain of Monascus is selected from the following One or more of the three strains: Aspergillus oryzae, the preservation number is CGMCC No. 0315; the red mold fungus, the preservation number is CGMCC No. 0316; the koji koji, the deposit number is CGMCC No. 0317, Fermentation culture at 15 to 35 ° C for more than 9 days, the strain can be cultured into a liquid strain or a solid strain. (3), the culture is sterilized at a temperature of 60 to 121 ° C, and the pressure is reduced by 60. Drying at ~80 °C to obtain functional red yeast rice.

The preparation method of the pharmaceutical composition of the present invention may also be the following method:

Preparing red yeast extract I and evening primrose extract II, and extracting extract I and extract II according to a conventional method, and adding a conventional adjuvant to prepare a clinically acceptable oral preparation, including but not limited to pills, powders, tablets, Granules, capsules, oral liquid preparations;

The red yeast extract I can be prepared by any of the following methods:

A, red koji 1 part by weight, each time adding 2~10 parts by volume of 50~90% ethanol, methanol or ethyl acetate, heating under reflux for 1-3 hours, extracting 2~3 times; extracting the filtrate, combining the filtrate, recycling The solvent is concentrated to 55~60 ° C to measure the relative density of 0. 95~1. 06 thick paste, to obtain extract I;

B, red koji 1 part by weight, each time adding 2~10 parts by volume of 50~90% ethanol, methanol or ethyl acetate heated under reflux for 1~3 hours, extracting 2~3 times; extracting the filtrate, combining the filtrate, recovering the solvent 5〜2. 0倍脱离子水混合,, room temperature or refrigerated place 2~, concentrated to 55~60 ° C, the relative density of 0.95~1.06 thick paste, concentrated in 0. 5~2. 1 hour, centrifugation, collecting precipitate, drying, to obtain extract I;

The evening primrose extract II is prepared by any of the following methods:

(1), taking the moon to see 1 part by weight of the herbal material, adding 4 to 20 parts by volume of water for steam distillation l~8h; to obtain extract II.

(2), the evening primrose is pulverized into a coarse powder, passed through a 10 to 80 mesh sieve, and placed in an extraction vessel for extraction, the extraction pressure is 7. 0~45Mpa, the extraction temperature is 30 to 80 ° C, and the extraction time is 30 min to 8 h. The extracting supercritical CO 2 is introduced into the separation vessel for analytical separation, and the analytical pressure is 4. 0~7. OMpa, the desorption temperature is 20 to 80 ° C, and the analysis time is 30 min to 8 h, and the extract II is obtained.

(3), smash the evening primrose into a coarse powder, add 3~15 parts by volume of 10~100% ethanol each time for 1~3 hours, extract 2~4 times, combine the extracts, recover the solvent, and concentrate A thick paste having a relative density of 0.95 to 1.06 was measured at 55 to 60 ° C to obtain Extract II.

Preferably, the method A described in the preparation method of the present invention comprises: taking 1 part by weight of red yeast medicine, adding 3 parts by volume of 75% ethanol to reflux for 3 hours, and extracting the extract; and adding the residue to 2 parts by volume of 75% ethanol for reflux for 2 hours. The extract is filtered, the filtrate is combined twice, the ethanol is recovered under reduced pressure, and concentrated to 55~60 ° C to measure the relative density of 0.95~1.06 thick extract to obtain extract I;

The method B described in the preparation method of the present invention is preferably: 1 part by weight of Monascus var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. var. Filtration, combining two filtrates, recovering ethanol under reduced pressure, concentrating to 55~60 ° C to measure the relative density of 0.95~1.06 thick paste, adding 1 times deionized water to the concentrate, mixing, room temperature or refrigerating for 8 hours . After centrifugation, the precipitate was collected, dried at 80 ° C for 8 small tests, and pulverized through a 60 mesh sieve to obtain extract I.

The evening primrose extract II described in the preparation method of the present invention is preferably prepared by the following method:

(1), taking 1 part by weight of the herbal material, adding 12 parts by volume of water for steam distillation for 4 hours; extract II.

(2) The evening primrose is pulverized into coarse powder, passed through a 40 mesh sieve, and placed in an extraction kettle for extraction. The extraction pressure is 25 MPa, the extraction temperature is 40 ° C, the extraction time is 4 h, and the supercritical CO 2 dissolved with the extract enters. The separation was carried out in a separation vessel, and the pressure was 5.0 Mpa, the desorption temperature was 50 ° C, and the analysis time was 4 h to obtain extract II.

(3), pulverize evening primrose into coarse powder, add 10 parts by volume of 50% ethanol for reflux for 2 hours, extract 3 times, combine the extracts, recover the solvent, and concentrate to 55~60 °C to measure the relative density. A thick paste of 0.95 to 1.06 gives Extract II.

The relationship of parts by weight per part by volume of the present invention is: gram per milliliter. The red yeast rice of the present invention may be a common red yeast medicine, or may be a commercially available functional red yeast rice or a high content red yeast rice.

The pharmaceutical composition of the present invention, red yeast and evening primrose, is compatible with each other, and synergistic effect is produced. With dehumidification, blood circulation The effect of phlegm, spleen and digestion. The pharmaceutical composition of the present invention is capable of lowering total cholesterol in blood, increasing high-density lipoprotein, lowering low-density lipoprotein content, and particularly exhibiting a lowering of triglyceride, and can significantly treat hypertriglyceridemia. The preparation process of the invention is reasonable, which not only improves the content of the active ingredient lovastatin in the red yeast rice, but also retains the active ingredients of each drug in the prescription, and fully exerts the effects of each drug. The preparation of the invention has the content of lovastatin as a quality control index, and the preparation quality is stable and controllable.

The effects of the present invention are demonstrated by pharmacodynamic tests below.

Test Example 1 Effect of the pharmaceutical composition of the present invention on hypertriglyceridemia in mice fed a high-fat word feeding model

1. Experimental materials:

(1) Preparation of test drugs:

Red yeast extract preparation: The preparation method is the same as the preparation method of extract I in Example 4.

Preparation of evening primrose extract: The preparation method is the same as the preparation of extract II in Example 4.

Preparation of compound medicine: See Example 4 for the preparation method.

All the above drugs were prepared by adding 1% CMC before use to make lml (liquid) I (raw drug).

(2) Preparation of high-fat emulsion: cholesterol 10g, lard 40g, sodium cholate 2g, propylthiouracil lg, Tween-80 lml, propylene glycol 15ml, distilled water added to 100ml

(3), reagent: Triglyceride (TG) kit, Beijing Zhongsheng Beikong Biotechnology Co., Ltd.

(4) Animals: ICR mice, male, weighing 25~30g, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

(5), instrument: Versa max tunable, Molecular Devices, centrifuge: (Anke LXJ-

Π Β).

2, methods and results

(1), Methods: 60 ICR mice were taken and served for 5 days. After confirming the health, the mice were randomly divided into 6 groups, 10 in each group. Blank control group, high fat model group (ig high fat emulsion 20ml/kg. d water), positive control group (ig high fat emulsion 20ml/kg. d ten fenofibrate 300mg/kg), red yeast group (ig High fat emulsion 20ml/kg. d + red yeast concentrate), evening primrose group (ig high fat emulsion 20ml/kg. d October primrose concentrate), compound group (ig high fat emulsion 20ml/kg. d + compound concentrate). At 8:30 am, give high-fat milk and its blank solution; at 15:30 in the afternoon, give water, positive control drugs, and each test drug. The cells were administered continuously for 20 days, and blood was taken at the 21st day. Plasma was prepared by anticoagulation with EDTA. The triglyceride content was determined by the kit method.

(2), the results are shown in Table 1.

Table 1 Effect of Chinese herbal compound composition on TG in high fat mice (X Shi SD, mg/dl) Group _t , high fat milk volume (ml/kg) n (only) triglyceride (mg/dl)

(g original medicine / kg body weight) Blank control group 20 (water) 10 110±22.3 high fat model group 20 10 195±43.5 ΔΔ positive control group 300 mg/kg 20 10 104±24.1**

Red yeast group 20 20 10 141±34.5* evening primrose group 20 20 10 170±45.1 compound group 20 20 10 120±30.5** 八八表示 compared with blank control group (t test), P<0.01; *, * * indicates comparison with the high fat model group (t test), P < 0.05, 0.01

Red yeast rice, evening primrose and compound can all reduce the serum TG content in high fat milk model mice. However, in the same dose, the red yeast group was 28% lower than the model group, the evening primrose group was 13% lower than the model group, and the compound group was 38% lower than the model group. The compound group mice had a greater TG reduction than the red yeast group. Group, evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 2 Effect of the pharmaceutical composition of the present invention on hypertriglyceridemia in a mouse fed with high fat vocabulary

1. Experimental materials:

(1) Preparation of test drugs:

Red yeast extract preparation: The preparation method is the same as the preparation method of extract I in Example 5.

Preparation of evening primrose extract: The preparation method is the same as the preparation of extract II in Example 5.

Preparation of compound medicine: See Example 5 for the preparation method.

All the above drugs were prepared by adding 1% CMC before use to make lml (liquid) / g (raw drug).

(2) Preparation of high fat emulsion: Same as Experimental Example 1.

(3), reagent: same as experimental example 1.

(5), Instrument: Same as Experimental Example 1.

2, methods and results

(1), Method: Same as Experimental Example 1.

(2) The results are shown in Table 2.

Table 2 Effect of Chinese herbal compound composition on TG in high fat mice (X Shi SD, mg/dl) Group _t , high fat milk volume (ml/kg) n (only) triglyceride (mg/dl)

(g original medicine / kg body weight)

Blank control group - 20 (water) 10 110 ± 22.3 high fat model group - 20 10 195 ± 43.5 Δ Δ positive control group 300 mg / kg 20 10 104 ± 24.1**

Red yeast group 20 20 10 137 ±35.7* evening primrose group 20 20 10 167±49.5 Compound group 20 20 10 124±32.6**

Eighth eight compared with the blank control group (t test), P <0.01; *, ** indicates comparison with the high fat model group (t test), P <0.05, 0.01

Red yeast rice, evening primrose and compound can all reduce the serum TG content in high fat milk model mice. However, in the same dose, the red yeast group was 30% lower than the model group, the evening primrose group was 14% lower than the model group, and the compound group was 36% lower than the model group. The TG of the compound group was lower than that of the red group. Group, evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 3 Effect of the pharmaceutical composition of the present invention on hypertriglyceridemia in a high fat diet-fed model mouse

1. Experimental materials:

(1) Preparation of test drugs:

Red yeast extract preparation: The preparation method is the same as the preparation method of extract I in Example 6.

Preparation of evening primrose extract: The preparation method is the same as the preparation of extract II in Example 6.

Preparation of compound medicine: See method example 6

All the above drugs were prepared by adding 1% CMC before use to make lml (liquid) lg (raw drug).

(2) Preparation of high fat emulsion: Same as Experimental Example 1.

(3), reagent: same as experimental example 1.

(5), Instrument: Same as Experimental Example 1.

2, methods and results

(1), Method: Same as Experimental Example 1.

(2) The results are shown in Table 3.

Table 3 Effect of Chinese herbal compound composition on TG in high-fat mice (X±SD, mg/dl) Group High-fat milk volume (ml/kg) n (only) Triglyceride (mg/dl)

(g original medicine / kg body weight)

Blank to control group 2200 ((water)) 1100 111100±±2222..33

High fat model group 20 10 195±43.5ΔΔ

Positive control group 300mg/kg 20 10 104±24.1**

Red song group 20 20 10 130±31.9*

Evening primrose group 20 20 10 183±42.5

Compound group 20 20 10 115±30.2** Eight-eighth comparison with blank control group (t test), P<0.01; *, ** indicates comparison with high-fat model group (t test), P<0.05, 0.01

Red yeast rice, evening primrose and compound can reduce the serum TG content in high fat milk model mice. But at the same dose In the case, the red yeast group was reduced by 33% compared with the model group, the evening primrose group was reduced by 6% compared with the model group, and the compound group was reduced by 41% compared with the model group; the TG reduction of the compound group was greater than that of the red yeast group and the evening primrose group. . It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 4 Experimental study on the hypolipidemic effect of the pharmaceutical composition of the present invention

1. Experimental materials:

(1), drugs

Preparation of compound medicine: See Example 4 for the preparation method.

Each of the above drugs was prepared by adding 1% CMC to 1 ml (liquid) / _g (raw drug).

(2) Animals: Wistar male rats weighing 150-200 g. Purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

(3), high fat feeding: cholesterol 2%, sodium cholate [pig] 0.5%, lard 15% (home made), propyl thiouracil 0.2%, basal feed powder 82.3%. The lard was melted in a 40 ° C water bath. Sodium cholate, cholesterol, and propylthiouracil are pulverized and mixed with a pulverizer, and the base is mixed, and then the melted lard is added and mixed well. Add a total of lOOOOg of raw material to 50ml of water and mix well. Press it into a block with a dough press (one cake per 30 g of mixed material). Dry and sterilize to get high fat.

(4), reagent: Triglyceride (TG) kit, Beijing Zhongsheng Beikong Biotechnology Co., Ltd.

(5), Instruments: Instruments: Versa max tunable, Molecular Devices, Centrifuge: (Anke LXJ-IIB).

2, methods and results

(1), hyperlipidemia modeling:

Fifty rats were divided into a blank control group (10) and a high-fat model (40), which were given normal basal support and high-fat support for 4 weeks. Fasting on the evening of the 28th, the next morning to cut the tail to take blood. Plasma was prepared by anticoagulation with EDTA. The triglyceride content was determined by the kit method. The results are shown in Table 2.

(2), Method: The normal basal rats were fed as a blank control group: continue to feed the normal basal feeding, and 1% CMC aqueous solution was administered by daily gavage; the high-fat model animals were randomly divided into 4 groups according to the blood lipid TG level. , high-fat model group: fed high-fat diet, daily administration of 1% CMC aqueous solution; red yeast group: daily administration of red yeast extract, the dose is 20g (original medicine) / kg body weight; evening primrose Group: The evening primrose extract was administered by daily gavage, and the dosage was 20 g (original medicinal material) / kg body weight; Compound group: The traditional Chinese medicine compound extract of the present invention was administered by gavage every day, and the dosage was 20 g (original medicinal material) /kg body weight. Continuous feeding for 4 weeks, fasting for 12 hours after the last administration, and taking blood at the end of the next morning. Plasma was prepared by anticoagulation with EDTA. The triglyceride content was determined by the kit method.

(3) The results are shown in Table 4.

Table 4 Effect of Chinese herbal compound composition on blood lipid content (X Shi SD, mg/dl)

Dosage

Group ((g original drug / kg body weight) ^η (only) triglyceride ( ^m g ^{/ dl)} blank control group - 10 93 ± 23.5

High fat model group - 10 160 ± 45.7 ΔΔ

Red yeast group 20 10 120±33.9* evening primrose group 20 10 143 ±37.1

Compound group 20 10 97 ± 25.4 ** VIII indicates comparison with blank control group (t test), P < 0.01; *, ** indicates comparison with high fat model group (t test), P < 0.05, 0.01

The results showed that red yeast rice, evening primrose and compound could reduce the serum TG content in high TG model rats induced by high fat feeding. Compared with the model group, the difference was significant P<0.05 and extremely significant difference. <0.01. From the TG reduction percentage: In the same dose, the red yeast group was reduced by 25% compared with the model group, the evening primrose group was reduced by 11% compared with the model group, and the compound group was reduced by 39% compared with the model group; The reduction was greater than that of the red yeast group and the evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 5 Experimental study on the hypolipidemic effect of the pharmaceutical composition of the present invention

1. Experimental materials:

(1), drugs

Preparation of compound medicine: See Example 5 for the preparation method.

(3), high fat feed: the same as experimental example 4.

(4), reagent: same as experimental example 4.

(5), Instrument: Same as Experimental Example 4.

2, methods and results

(1), hyperlipidemia modeling: the same as experimental example 4.

(2) Experimental method: Same as Experimental Example 4. (3), the results are shown in Table 5

Table 5 Effect of Chinese herbal compound composition on blood lipid content (X±SD, mg/dl) Group ((g original drug Z material/kg body weight) ^η (only) triglyceride ( ^mg/dl )

Blank control group - 10 93 ± 23.5

High fat model group - 10 160 ± 45.7 ΔΔ

Red yeast group 20 10 114±31.7* evening grass group 20 10 150±36.4

Compound group 20 10 95±24.8**

The results showed that red yeast rice, evening primrose and compound could reduce the serum TG content in high TG model rats induced by high fat feeding. Compared with the model group, the difference was significant P<0.05 and extremely significant difference. <0.01. From the percentage of TG reduction: At the same dose, the red yeast group was 29% lower than the model group, the evening primrose group was reduced by 6% compared with the model group, and the compound group was 41% lower than the model group; The reduction was greater than that of the red yeast group and the evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 6 Experimental study on the hypolipidemic effect of the pharmaceutical composition of the present invention

1. Experimental materials:

(1), drugs

Preparation of compound medicine: See Example 6 for the preparation method.

(3), high fat feed: the same as experimental example 4.

(4), reagent: same as experimental example 4.

(5), Instrument: Same as Experimental Example 4.

2, methods and results

(1), hyperlipidemia modeling: the same as experimental example 4.

(2) Experimental method: Same as experimental example 4.

(3), the results are shown in Table 6

Table 6 Effect of Chinese herbal compound composition on blood lipid content (X±SD, mg/dl) Group ((g original drug Z material / kg body weight) ⁿ (only) triglyceride (mg ^{/ dl} ) blank control group - 10 93 ± 23.5

High fat model group - 10 160 ± 45.7 ΔΔ

Red yeast group 20 10 110±30.1* evening primrose group 20 10 155±37.2

Compound group 20 10 101±25.7**

The results showed that red yeast rice, evening primrose and compound could reduce the serum TG content in high TG model rats induced by high fat feeding. Compared with the model group, the difference was significant P<0.05 and extremely significant difference. <0.01. From the percentage of TG reduction: At the same dose, the red yeast group was 31% lower than the model group, the evening primrose group was reduced by 3% compared with the model group, and the compound group was reduced by 37% compared with the model group; The reduction was greater than that of the red yeast group and the evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood. Test Example 7 Experimental study on the hypolipidemic effect of the pharmaceutical composition of the present invention

1. Experimental materials:

(1), drugs

Preparation of red yeast extract: 1 part by weight of Hongqu and 1 part by weight, added with 6 parts by volume of 75% ethanol, and refluxed for 3 hours, and the extract was filtered; the residue was added to 6 parts by volume of 75% ethanol and refluxed for 2 hours, and the extract was filtered. The filtrate was combined twice, and ethanol was recovered under reduced pressure, and concentrated to 55 to 60 ° C to measure a relative density of 0.95 to 1.06. dry.

Preparation of evening primrose extract: 1 part by weight of herbal material, 6 parts by volume of 75% ethanol was added for reflux for 3 hours, and the extract was filtered; the residue was added to 6 parts by volume of 75% ethanol and refluxed for 2 hours, and the extract was filtered. The filtrate was combined twice, and ethanol was recovered under reduced pressure, and concentrated to 55 to 60 ° C to measure a relative density of 0.95 to 1.06. dry.

Preparation of compound medicine: See Example 7 for the preparation method.

(3), high fat feed: the same as experimental example 4.

(4), reagents: triglyceride (TG), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (LDL-C), using Beijing Zhongshengbei control organism Technology Co., Ltd. kit determination.

(5), instrument: Versa max tunable, Molecular Devices, centrifuge: (Anke LXJ-II Β).

2, methods and results (1) Modeling for hyperlipidemia: Same as Experimental Example 4.

(2) Experimental method: Same as experimental example 4.

(3), the results are shown in Table 7

Table 7 Effect of Chinese herbal compound composition on blood lipid content (X±SD)

Dosage dose Triglyceride TC HDL-C LDL-C group n (only)

(g original drug kg body weight) (mg / dl) (mg / dL) (mg / dL) (mg / dL) blank control group 10 104 101.74 ± 16.25 45.11 ± 9.33 62.61 ± 16.72 high fat model group 10 173 ± 43.1 ^ΔΔ 234.82±23.60 ^ΔΔ 28.39±9.06 ^Δ 202.61 ±22.94 ^Δ红曲组20 10 119±33.0** 140.04± 15.09** 40.83±8.17* 107.33 ±15.94** Evening primrose group 20 10 170±40.2 228.63 ±29.40 32.67 ± 8.56 195.61±22.17 Compound group 20 10 102±29.6** 132.30±20.12** 43.56±11.28* 100.72± 10.50**

△, 八八表示 compared with the blank control group (t test), P <0.05, P <0.01;

*, ** indicates comparison with the high-fat model group (t test), P < 0.05, P < 0.01.

The results showed that red yeast rice, evening primrose and compound could reduce the serum TG content in high TG model rats induced by high fat feeding. Compared with the model group, the difference was significant P<0.05 and extremely significant difference. <0.01. From the TG reduction percentage: In the same dose, the red yeast group was 31% lower than the model group, the evening primrose group was 2% lower than the model group, and the compound group was 41% lower than the model group; The reduction was greater than that of the red yeast group and the evening primrose group. It shows that the traditional Chinese medicine compound composition has a more obvious effect of lowering TG in blood.

From the results of TC, HDL-C, and LDL-C, the evening primrose group had no significant effect on reducing TC, LDL-C, and increasing HDL-C, but it had a certain effect. After compounding with red yeast, it enhances the reduction of TC, LDL-C and the effect of HDL-C. The combination of the two drugs has a synergistic effect. Experimental Example 8 Formulation screening test

1. Tested drugs: Water extracting and alcohol extracting sites, two monomeric compounds, and four effective extracts of 25 kinds of medicinal materials were screened, and one Chinese herbal medicine preparation and red yeast extract prepared on the market were compared. Into the compound. See Table 8 for details. Among them, all the medicinal materials were tested for the combination of the water extract and the red yeast extract compound, the ethanol extract and the red yeast extract.

2, preparation method:

(1) Preparation of extract when A is a medicinal material:

• Water extraction method: A medicinal material 100g plus 8 times the amount of water, soaked for 30min, heated to boiling and kept slightly boiling for 30min;

100 mesh sieve filtration; filter residue added 5 times the amount of water, heated to boiling and kept slightly boiling for 20 min _; 100 mesh sieve filtration; the filtrate was combined, concentrated, and dried to obtain a water extract of the medicinal material.

• Ethanol extraction method: A medicinal material 100g plus 8 times 75% ethanol, soaked for 30min, refluxed for 30min; 100 The sieve was filtered; the residue was further added with 5 times of 75% ethanol, refluxed for 20 min; 100 mesh sieve; the filtrate was combined, concentrated, and dried to obtain an ethanol extract of the drug.

(2) Preparation of red yeast extract of B medicine: Take 100g of red yeast medicine, add 3 times 75% ethanol and reflux for 3 hours, and extract the extract; filter residue is added to 2 times 75% ethanol for reflux for 2 hours, and the extract is filtered. The filtrate was combined twice, and the ethanol was recovered under reduced pressure, concentrated, and dried to give a red yeast extract.

(3), preparation method of compound:

The water extract of the A medicinal material prepared by the method of 2. (1) and the red yeast extract are combined to form a compound; or the ethanol extract of the medicinal material and the red yeast extract are combined to form a compound. Prepare 100 ml of 0.5% aqueous sodium carboxymethylcellulose solution immediately before use.

(4), a method for preparing a test compound when A is a commercially available extract

Take 25g of extract and add the extract of B medicine. (For the extraction method of B medicine, see 2. (2) above). Before use, 100 ml of 0.5% sodium carboxymethylcellulose solution was prepared.

(5), a method for preparing a test compound when A is a commercially available monomer

Commercially available monomers curcumin or take matrine 3g (600mg / kg dose group), lg (200mg / kg dose _{group), 0.5g (100m g / k} g dose group), B is added herbs extracts (B For the extraction method of medicinal materials, see 2. (2) above. 100 ml was prepared with 0.5% aqueous sodium carboxymethylcellulose solution immediately before use.

(6), a method for preparing a test compound when A is a commercially available preparation

Take 75 tablets of Gynostemma pentaphyllum (equivalent to 1500 mg of Gynostemma pentaphyllum) and add the extract of B. (The extraction method of B drug is shown in 2. (2) above). Prepare 100 ml of 0.5% aqueous sodium carboxymethylcellulose solution immediately before use.

3, the experimental drug dose: are the following doses

(1) When A is a medicinal material, the dose of the compound drug to be tested is:

The doses of A and B herbs are 20 g of medicinal material / k _g body weight (the concentration is 1 g of medicinal material / m 1 ), which is equivalent to 10 times of clinical dosage;

(2) When A is a commercially available extract (such as Salvia miltiorrhiza, Seabuckthorn, Ginkgo biloba), the dose of the compound to be tested is: A dose of 5 g of extract I kg body weight, dose of B containing drug is 20 g medicinal material / kg body weight (prepared concentration: compound containing A extract 0.25 g / m 1, containing Bl g medicinal material / m 1);

(3) When A is a commercially available monomer (such as curcumin or matrine), the dose of the compound to be tested is:

The dose containing A is 600~100mg monomer/kg body weight, and the dose containing B medicine is 20g medicine/kg body weight (preparation concentration: compound A contains 30mg / m 1~5mg/ml, contains Bl g medicine / m 1 )

(4) When A is a commercially available preparation, the dose of the compound to be tested is: The dose containing the drug A is 300 mg/kg body weight, and the concentration is 15 mg/ml; the dose containing the drug B is 20 g drug material I kg body weight (the concentration is 1 g drug/m 1 ).

4. Model establishment:

(1) Fructose-induced high TG mouse model:

Male ICR mice, about 30 g. Mice were dosed without fasting. The blank group and the model group were intragastrically administered with a 0.5% aqueous solution of sodium carboxymethylcellulose in a volume of 20 ml/kg, and the fenofibrate group (300 mg/kg) was given fenofibrate in an equal volume. Continuous administration for 3 days. On the 2nd to 3rd day, the water was forbidden for 24 hours, and the drinking water was replaced with 20% fructose solution for 20 hours. The blood was cut at 20 hours after administration. TG was measured. Plasma was prepared by anticoagulation with EDTA (centrifugation 2400 rpm X 10 min). The TG value was measured.

(2) Triton WR-1339 high TG mouse TG model

Triton WR-1339 (Sigma): A concentration of 50 mg/mL was prepared using 0.9% sodium chloride solution. It was administered at a dose of 250 mg/kg.

Male ICR mice, about 30 g. The blank group and the model group were administered with 0.5% CMC solution, the fenofibrate group was administered at a dose of 300 mg/kg, and the test drug was administered at a dose of 20 g/kg. TG was measured 2 days after continuous administration of i. g. On the third day, blood was taken from each group, i. g. was administered, i.v. was modeled (Triton 0. 05 ml/10 g was injected into the tail vein, and physiological saline was injected into the blank group), and blood was taken at 24 h after modeling to determine the TG value.

(3) Effects of TG on multiple doses of normal mice

Male ICR mice, about 30g. Mice were dosed without fasting. Blank control group to 20ml volume / k _g of orally administered 0.5% aqueous sodium carboxymethyl cellulose, fenofibrate positive drug group (300mg / kg) and each test drug administration group in equal volume of fenofibric Specifically, continuous administration for 4 days, blood was taken at the 5th tail. Plasma was prepared by anticoagulation with EDTA. The TG value was measured.

5. Mode of administration: i. g

6. Dosing volume: 20m 1 / k _g

7. Positive control drug: fenofibrate 300m g / k g body weight

8. Mathematical statistics: t test, P < 0. Q 5 has significant difference, P < 0. Q 1 has extremely significant difference

9. Results: See Table 4

Table 8 preliminary screening of drug grouping

vine

Salvia miltiorrhiza extract compounded extract **

Seabuckthorn Extract Compound Commercial Extract

Curcumin compound Commercial monomer * * Matrine compound Commercial monomer Toxicity Toxicity Gynostemma total glycoside tablet compound Commercial preparation

Note:

* indicates that TG was significantly lower than the model control group (P < 0.05);

** indicates that compared with the model control group, TG has a significant decrease (P<0.01);

"There was a trend" indicating that the TG value was lower than that of the model control group, but the t test was not statistically significant, indicating that the drug had a decreasing trend.

(1) Fructose model: Each group was compared with the model control group and repeated experiments were performed. The experimental results are summarized as follows:

**: Salvia miltiorrhiza, Gynostemma pentaphyllum, Alisma, Rhizoma Curcumae, Licorice, Hawthorn, Chuanxiong;

*: Curcumin, Seabuckthorn, Evening Primrose Oil, Shayuanzi, Sophora flavescens;

There are trends: Ginkgo biloba, Atractylodes, Sedum

(2), Triton model: Each group was compared with the model control group, and repeated experiments were carried out. The experimental results are summarized as follows:

**: turmeric, Sophora flavescens, Alisma, Rhizoma, Yujin

*: Gynostemma, Evening Primrose Oil, Pueraria,

There are trends: Ginkgo biloba, cooked rhubarb, sea buckthorn, milk thistle, capillaris, hawthorn, Chuanxiong, red oak

(3) After repeated administration of normal mice, there are extremely significant reductions in TG: 莪2, the second stage is to screen the above preferred drug regrouping (administration method, dosage, etc. are the same) See Table 9)

Table 9 Summary of Compound Remarks for Drug Regrouping Screening

Pueraria Compound Ethanol* *

See you in the moon

Evening Primrose Oil Complex * *

Grass oil

From the above results, it can be seen that different drug combinations show different results in reducing TG in various models, possibly due to different mechanisms of action. However, the end result is that it shows a better effect of reducing TG.

detailed description:

Example 1: Preparation of the pharmaceutical composition dropping pellet of the present invention

6 kg of ordinary red yeast rice was added, and 24 L of 80% ethyl acetate was added and heated under reflux for 2 hours, and extracted three times; the extract was filtered, the filtrate was combined, the solvent was recovered, and concentrated to a thick paste to obtain extract I.

Take the moon to see the herbs lkg: add 20L of water for steam distillation, collect the distilled evening primrose oil, that is, the extract

II.

The extracts I and II are added to the preparation auxiliary materials, and the clinically accepted dropping pills are prepared according to a conventional process. Example 2: Preparation of a pharmaceutical composition tablet of the present invention

Take 4kg of functional red yeast rice, add 8L of 90% ethanol and heat to reflux for 1 hour, and filter; add 8L of 90% ethanol and heat to reflux for 1 hour, and filter; combine the filtrate, recover the solvent, concentrate to thick paste, and obtain extract I.

The herbs were pulverized into coarse powder and placed in an extraction kettle for extraction. The extraction pressure was 30 MPa, the extraction temperature was 35 ° C, the extraction time was 2 h, and the supercritical solution of the extract was dissolved (: 0 ₂ into the separation kettle for analysis) Separation, analytical pressure 6. OMpa, desorption temperature 40 ° C, analysis time 30 min, extract II.

Extracts I and II were added to the preparation excipients, and clinically accepted tablets were prepared according to a conventional procedure. Example 3: Preparation of oral liquid of pharmaceutical composition of the present invention

Take 1kg of red yeast medicine, add 4L of 50% ethanol and reflux for 3 hours, and extract the extract; the filter residue is added to 2L of 50% ethanol for reflux for 3 hours, and the extract is filtered. The filtrate was combined twice, and ethanol was recovered under reduced pressure, and concentrated to 55 to 60 ° C to measure a relative density of 0.95 to 1.06. Add 0.2 volume of deionized water to the concentrate and mix for 12 hours. After centrifugation, the precipitate was collected to obtain a red yeast extract I.

Take 6kg of herbal medicine, add 10 times of 95% ethanol reflux extraction 1. 5h, filter; add 10 times 95% ethanol reflux extraction 1. 5h, filter; combine the filtrate, concentrate, get extract II.

Extracts I and II were added to the preparation excipients, and a clinically accepted oral solution was prepared according to a conventional procedure. Example 4: Preparation of granules of the pharmaceutical composition of the invention Take ordinary red yeast lkg, add 4L of 80% ethanol each time and heat to reflux for 2 hours, extract 2 times; extract the filtrate, combine the filtrate, recover the solvent, concentrate to thick paste, and obtain extract I.

Take the moon to see the herbs lkg: add 10L of water for steam distillation, collect the distilled evening primrose oil, that is, the extract

II.

The extracts I and II are added to the preparation excipients, and the clinically accepted granules are prepared according to a conventional process. Example 5: Preparation of the pharmaceutical composition capsule of the present invention

2 kg of functional red yeast rice was added, and 6 L of 70% ethanol was added and heated under reflux for 3 hours, and filtered; 4 L of 75% ethanol was further added and heated under reflux for 2 hours, and filtered; the filtrate was combined, the solvent was recovered, and concentrated to a thick paste to obtain an extract I.

Take the moon and see the herbal powder lkg into coarse powder, pass through a 20 mesh sieve, and put it in the extraction kettle for extraction. The extraction pressure is 20Mpa, the extraction temperature is 40 °C, the extraction time is 3h, and the supercritical ∞ ₂ dissolved in the extract enters. Analytical separation was carried out in a separation vessel, and the pressure was 6.0 Mpa, the desorption temperature was 45 ° C, and the analysis time was 30 min to obtain extract II.

The extracts I and II are added to the preparation excipients, and the clinically accepted capsules are prepared according to a conventional process. Example 6: Preparation of the pharmaceutical composition capsule of the present invention

Take the functional red yeast medicine lkg, add 3L75% ethanol reflux extraction for 3 hours, the extract is filtered; the filter residue is added to 2L75% ethanol reflux extraction for 2 hours, and the extract is filtered. The filtrate was combined twice, and ethanol was recovered under reduced pressure, and concentrated to 55 to 60 ° C to measure a relative density of 0.95 to 1.06. Add 0.2 volume of deionized water to the concentrate and mix for 8 hours. After centrifugation, the precipitate was collected to obtain extract I.

Take the moon to see the herbs 6k _g: add 10 times the volume of 75% ethanol and reflux for 2 times, the extraction time is lh. The filtrate was separately filtered, and the filtrate was combined and concentrated under reduced pressure to give the extract II.

The extracts I and II are added to the preparation excipients, and the clinically accepted capsules are prepared according to a conventional process. Example 7: Preparation of the pharmaceutical composition capsule of the present invention

Take the red yeast medicinal material lkg, the moon-seeing herbal material lkg into 6L75% ethanol reflux extraction for 3 hours, the extract is filtered; the filter residue is added to 6L75% ethanol reflux extraction for 2 hours, and the extract is filtered. The filtrate was combined twice, and the ethanol was recovered under reduced pressure, concentrated, and dried. The preparation auxiliary is added, and a clinically accepted capsule is prepared according to a conventional process.

Example 8: Preparation of the pharmaceutical composition of the invention

Take the red yeast medicinal material lkg, the moon-seeing herbal material lkg into 6L75% ethanol reflux extraction for 3 hours, the extract is filtered; the filter residue is added to 6L75% ethanol reflux extraction for 2 hours, and the extract is filtered. The filtrate was combined twice, and the ethanol was recovered under reduced pressure, concentrated, and dried. The preparation auxiliary is added, and a clinically accepted soft capsule is prepared according to a conventional process.

Claims

Claim

A pharmaceutical composition having a blood lipid regulating effect, characterized in that the pharmaceutical composition of the pharmaceutical composition is: 1 part by weight of red yeast 1 part by weight of evening primrose.

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition of the pharmaceutical composition is any one of the following:

Red yeast 2 parts by weight evening primrose 4 parts by weight;

Red yeast 4 parts by weight evening primrose 2 parts by weight;

Red yeast 1 part by weight evening primrose 4 parts by weight;

Red yeast 4 parts by weight Evening primrose 1 part by weight.

The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is produced by the following method: Step one, red yeast extract I preparation

Red yeast extract I is prepared by any of the following methods:

B, red koji 1 part by weight, each time adding 2~10 parts by volume of 50~90% ethanol, methanol or ethyl acetate heated under reflux for 1~3 hours, extracting 2~3 times; extracting the filtrate, combining the filtrate, recovering the solvent Concentrating to a thick paste of 0.55 to 1.06, and concentrating it to a concentration of 0.95 to 1.06, and adding 0. 5~2. 0 times deionized water to mix, room temperature or refrigerated for 2 to 1 hour, centrifugation, collection Precipitating, drying, obtaining extract I;

Step 2, evening primrose extract II preparation

Evening Primrose Extract II is prepared by any of the following methods:

(3), pulverizing evening primrose into coarse powder, adding 3~15 parts by volume of 10~100% ethanol each time for 1~3 hours, extracting 2~4 times, combining the extracts, recovering the solvent, and concentrating 55~60 ° C to measure the relative density of 0.95~1.06 thick paste, to obtain extract II;

Step three, preparation The extract I and the extract π are added to a conventional excipient according to a conventional method to prepare a clinically accepted pill, powder, tablet, granule, capsule or oral liquid preparation.

The pharmaceutical composition according to claim 3, wherein the method A in the first step is: taking 1 part by weight of the red yeast medicine, adding 3 parts by volume of 75% ethanol and refluxing for 3 hours, and extracting the extract; The residue was extracted by refluxing with 2 parts by volume of 75% ethanol for 2 hours, and the extract was filtered, and the filtrate was combined twice. The ethanol was evaporated under reduced pressure, and concentrated to a thick paste of a density of 0.95 to 1.06 at 55 to 60 ° C to obtain an extract I.

The pharmaceutical composition according to claim 3, wherein the method B in the first step is: 1 part by weight of red yeast medicine, and 3 parts by volume of 75% ethanol is added for reflux extraction for 3 hours, and the extract is filtered; Add 2 parts by volume of 75% ethanol and reflux for 2 hours, filter the extract, combine the two filtrates, recover the ethanol under reduced pressure, concentrate to 55~60 ° C to measure the relative density of 0.95~1.06 thick paste, add 1 to the concentrate Mix with deionized water and leave it at room temperature or refrigerate for 8 hours. After centrifugation, the precipitate was collected, dried at 80 ° C for 8 hours, and pulverized through a 60 mesh sieve to obtain Extract I.

6. The pharmaceutical composition according to claim 3, wherein the second step is any one of the following methods:

The method for preparing a pharmaceutical composition according to claim 1 or 2, wherein the method is as follows: Step 1: Preparation of Monascus extract I

Red yeast extract I is prepared by any of the following methods:

B, red koji 1 part by weight, each time adding 2~10 parts by volume of 50~90% ethanol, methanol or ethyl acetate heated under reflux for 1~3 hours, extracting 2~3 times; extracting the filtrate, combining the filtrate, recovering the solvent Concentrating to a thick paste of 0.55 to 1.06, and concentrating it to a concentration of 0.95 to 1.06. The mixture is added with a concentration of 0. 5~2. 0 times deionized water, mixed at room temperature or refrigerated for 2 to 12 hours, centrifuged, collected. Precipitating, drying, obtaining extract I;

Step 2, evening primrose extract II preparation

Evening Primrose Extract II is prepared by any of the following methods: (1), taking 1 part by weight of the herbal material, adding 4 to 20 parts by volume of water for steam distillation 1~8h; extract II.

(3), smash the evening primrose into a coarse powder, add 3~15 parts by volume of 10~100% ethanol each time for 1~3 hours, extract 2~4 times, combine the extracts, recover the solvent, and concentrate 55~60 ° C to measure the relative density of 0.95~1.06 thick paste, to get extract II; Step three, the preparation

The extract I and the extract II are added to a conventional excipient according to a conventional method to prepare a clinically accepted pellet, powder, tablet, granule, capsule or oral liquid preparation.

The method for preparing a pharmaceutical composition according to claim 7, wherein the methods for preparing the red yeast extract I in the A and B methods are:

Method A: 1 part by weight of red yeast medicine, 3 parts by volume of 75% ethanol was added to reflux for 3 hours, and the extract was filtered; the residue was added to 2 parts by volume of 75% ethanol and refluxed for 2 hours, and the extract was filtered, and the filtrate was combined twice. The ethanol was recovered by pressure, and concentrated to 55 to 60 ° C to measure a thick paste having a relative density of 0.95 to 1.06 to obtain extract I.

Method B: 1 part by weight of Monascus sinensis, 3 parts by volume of 75% ethanol was added for reflux for 3 hours, and the extract was filtered; the residue was added to 2 parts by volume of 75% ethanol and refluxed for 2 hours, and the extract was filtered, and the filtrate was combined twice. The ethanol was recovered, concentrated to 55~60 ° C to measure the relative density of 0.95~1.06 thick paste, and the concentrate was mixed with 1 times of deionized water, and allowed to stand at room temperature or refrigerated for 8 hours. After centrifugation, the precipitate was collected, dried at 80 ° C for 8 hours, and pulverized through a 60 mesh sieve to obtain Extract I.

The method for producing a pharmaceutical composition according to claim 7, wherein the evening primrose extract II is prepared by any one of the following methods:

The pharmaceutical composition according to claim 1 or 2, wherein the red yeast rice is ordinary red yeast or Functional red yeast.

The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is added to a conventional excipient according to a conventional method to prepare a clinically acceptable pill, powder, tablet, granule, capsule or oral liquid. preparation.

The use of the pharmaceutical composition according to any one of claims 1 to 6 for the preparation of a medicament for treating hyperlipidemia.

13. The use according to claim 12, wherein said treating hyperlipidemia means lowering blood triglyceride content.

14. The use according to claim 12, wherein said treating hyperlipidemia means lowering blood LDL content, and/or increasing blood high density lipoprotein content.

15. The use according to claim 12, wherein said treating hyperlipidemia means reducing the total cholesterol content in the blood.