CN1288747A - Pharmaceutical composition containing dihydromyricetin - Google Patents

Abstract

The invention discloses a pharmaceutical composition and a health product containing dioxymyricetin, which have the effects of treating hepatitis, protecting the liver, resisting bacteria and reducing inflammation, relieving pain, lowering blood lipids, removing phlegm and enhancing the immunity of the immune system.

Description

Pharmaceutical composition containing dihydromyricetin

The invention belongs to the field of natural medicines for treating hepatitis, protecting liver and health care, relieving pain, lowering blood fat, eliminating phlegm, antibacterial and anti-inflammatory.

但是没有对二氢杨梅素的活性进行研究。The document Walter Karrerr, Birkhauser Verlag, Basselundstuttgart (1958), P.652, NO: 1640 discloses the structural formula of dihydromyricetin (Ampelopsin),

However, no studies have been conducted on the activity of dihydromyricetin.

The applicant surprisingly found that dihydromyricetin has the functions of treating hepatitis, protecting liver and health care, antibacterial and anti-inflammatory, analgesic, reducing blood fat, eliminating phlegm and enhancing immune system immunity. Dihydromyricetin can be mixed with pharmaceutically acceptable carriers or excipients to prepare pharmaceutical compositions. Dihydromyricetin can be obtained from grapes. It can also be obtained by conventional methods in the field of chemistry.

One object of the present invention is to provide a pharmaceutical composition containing dihydromyricetin and a pharmaceutically acceptable carrier or excipient for treating hepatitis.

Another object of the present invention is to provide an antibacterial and anti-inflammatory pharmaceutical composition containing dihydromyricetin and a pharmaceutically acceptable carrier or excipient.

Another object of the present invention is to provide a pharmaceutical composition and health product containing dihydromyricetin for liver protection and health care.

Another object of the present invention is to provide a pharmaceutical composition and a health care product containing dihydromyricetin for enhancing immunity of the immune system.

Another object of the present invention is to provide an anti-inflammatory and analgesic pharmaceutical composition or external medicine containing dihydromyricetin.

Another object of the present invention is to provide a pharmaceutical composition and health product containing dihydromyricetin for reducing blood lipid.

Another object of the present invention is to provide pharmaceutical compositions and health products containing dihydromyricetin for eliminating phlegm.

Another object of the present invention is the use of dihydromyricetin in the preparation of pharmaceutical compositions for treating hepatitis, hepatoprotection, anti-inflammation, analgesia, lowering blood fat, eliminating phlegm or improving immunity.

Dihydromyricetin can be obtained by decocting grape plants with various solvents (water and/or organic solvents), decocting one or more times, concentrating and cooling the decoction, standing overnight, and detecting the concentration of the precipitate and the supernatant. The medicinal ingredients are active precipitates, which are chromatographically separated to obtain crystals.

The principle of the invention is to trace the pharmacological activity of dihydromyricetin by establishing a pharmacological screening model, and to find its medicinal application. Experiments on the effect of carbon tetrachloride-induced experimental liver injury in mice and animal model experiments on the effect of D-galactosamine-induced experimental liver injury in mice show that dihydromyricetin has obvious hepatoprotective effects; Antibacterial and antiviral tests in vitro show that dihydromyricetin has antibacterial and anti-inflammatory effects; serum lectin tests show that dihydromyricetin has the effect of enhancing the immunity of the immune system.

It is found that dihydromyricetin can be used in the fields of pharmacy, health products and food.

Dihydromyricetin can be directly used as medicine or mixed with pharmaceutically acceptable carriers or excipients to form a pharmaceutical composition. Dihydromyricetin can also be used as an active ingredient in health care products. The composition of the present invention can also be formulated into beverages, etc. .

The pharmaceutical composition of the present invention can be prepared into a pharmaceutical composition by conventional preparation techniques and carriers in the pharmaceutical field. The composition of the invention can be prepared into tablets, granules, capsules and external medicines, and can also be added to tea or beverages to make health-care teas or beverages.

The common formula of the tablet can be dihydromyricetin: 10%-90% by weight, lactose 90%-10% by weight, 1.5g of sodium carboxymethylcellulose, 0.5g of magnesium stearate, and an appropriate amount of 50% ethanol. The general formula of the granule can be: 5-100% weight ratio of dihydromyricetin, (sucrose+dextrin) 95-0% weight ratio, sucrose:dextrin=2:1. The common formula of the capsule can be: dihydromyricetin 10-100% by weight, starch 90-0% by weight. However, those skilled in the art can also prepare other dosage forms with well-known techniques and carriers in the art as needed.

The invention is illustrated in detail by the following examples. However, it should be understood that these examples are only illustrative of the present invention, not limiting the scope of the present invention in any way.

Embodiment 1: tablet: dihydromyricetin 25g, lactose 53g, sodium carboxymethylcellulose 1.5g, magnesium stearate 0.5g, 50% ethanol 5ml

The above components are mixed in a mixer and compressed into tablets.

Embodiment 2: 1 part weight of dihydromyricetin syrup, 2.5 parts weight of sucrose, 1.25 parts weight of dextrin, 5 parts weight of ethanol

Mix the above ingredients to get a syrup.

Embodiment 3: electuary dihydromyricetin 115g, powdered sugar 345g, dextrin 145g, ethanol 5ml

The above components are mixed and then dried to obtain granules.

Embodiment 4: capsule dihydromyricetin 100g, starch 20g,

After the above components are mixed evenly, put into capsules. Example 5: Effect of dihydromyricetin on experimental liver injury in mice caused by carbon tetrachloride:

The animals were randomly divided into six groups: normal control group, positive control group, model control group and three groups of dihydromyricetin: 100mg/kg, 200mg/kg, 400mg/kg. The normal control group is that the animals are not given any medicine, only the solvent is given; the positive control group is bifendate 200mg/kg, qdx6, po, bifendate is a commonly used hepatitis treatment drug, and the model animals are treated with bifendate, Observing the curative effect, as a control; the model control group is to test the drug effect, the animal must be made into a hepatitis model first, so that the animal suffers from hepatitis; the dihydromyricetin group is based on the above model, and then given dihydromyricetin to observe the curative effect. All three groups are Bidx6 po. Animals in each group were administered in groups, 10 mice in each group, twice a day, taking a volume of 0.5ml/20g mice, and taking it for six consecutive days, intraperitoneally injecting 1% carbon tetrachloride oil solution 0.2 ml/mouse, after fasting for 16 hours, take blood from orbital vein, separate serum, measure serum SGPT (ALT), SGOT (AST) content and serum total bilirubin (T-BIL sodium benzoate caffeine colorimetric method) content , and compared with the positive control group, normal control group, and model control group, the model control group took the same amount of corresponding solvent, and the positive control group took bifendate 200mg/kg, pox6, once a day. The results are shown in Table 1.

Table 1. Changes of Dihydromyricetin in Serum SGOT of Mice Hepatic Injury Induced by Carbon Tetrachloride group Dose mg/kg Serum SGPT(U)X±SD Serum SGOT(U)X±SD T-BIL Corresponding solvent Model control group Corresponding solvent 109.2_9.8 83.7±4.6 5.78±2.91 Dihydromyricetin 50 Bidx6po 108.3_10.12 79.4±10.3 Dihydromyricetin 100 Bidx6po 85.3_4.34 71.0±10.4 4.24±2.11 Dihydromyricetin 200 Bidx6po 68.3_4.89** 68.4±9.2** 3.42±1.32 Bifendate 200 qdx6po 63.6_4.10** 61.2±4.2** 3.12±1.24 *P<0.05**P<0.01 Compared with the model group, it can be seen from the data in Table 1 that dihydromyricetin has an excellent effect on treating hepatitis. As well as reducing the content of T-BIL, it shows that dihydromyricetin has obvious hepatoprotective effect of lowering enzymes and reducing jaundice. Example 6: Protective effect of dihydromyricetin on D-galactosamine-induced liver injury in mice

The animals were randomly divided into six groups, the positive control group, the model control group, the normal control group, and the dihydromyricetin component were divided into three groups: 100mg/kg, 200mg/kg, 400mg/kg, the significance of each group is the same as the above-mentioned embodiment 4. The normal control group and the model control group took corresponding solvents, and the positive control group was intraperitoneally injected with 200mg/kg, pox6, qd, 1 hour after the last administration, D-galactosamine 100mg/kg, 16 hours later, blood was taken from the orbit, and the serum was separated , Determination of serum SGPT, changes in serum SGOT, the results are shown in Table 2

Table 2 Dihydromyricetin on the changes of serum SGPT and SGOT of D-galactosamine-induced liver injury in mice group Dose mg/kg Serum SGPT(U)XD Serum SGOT(U)XD normal control group Corresponding solvent <40 33.7_4.7 Model control group Corresponding solvent 90.7_17.3 58.3_15.5 Dihydromyricetin 50 Bidx6po 48 50 Dihydromyricetin 100 Bidx6po 34_12.8 41.2_11.0 Dihydromyricetin 200 Bidx6po 22.4** 31.8* Bifendate 200 qdx6po 23.6_3.9** 33.8_15.2* *P<0.05, Material P<0.01 Compared with the model group, it can be seen from the data in Table 2 that dihydromyricetin has an excellent hepatoprotective effect. Embodiment 7: Antibacterial and antiviral experiments of dihydromyricetin in vitro:

Isolation and Identification (Titration) of Influenza Virus

At present, chicken embryos are commonly used to isolate influenza virus. The route of inoculation is the allantoic cavity of chicken embryos. After 3 days, the lesion characteristics of post-chicken embryo death and hemagglutinin appear. For this, we use this method to detect dihydromyricetin Identification. We use a certain concentration of extract and 4 hemagglutination units of influenza virus to mix 1:1, let the two react at room temperature for 20 minutes and then inject chicken embryos, the injection volume is 0.2ml per embryo. In the experiment, chicken embryos aged 9 to 11 days were used to inoculate the allantoic cavity. After culturing for 48 hours, they were placed in the liquid at 4°C, and then the urine was collected aseptically. After being centrifuged at 1000rpm for 10 minutes, the supernatant was taken and packed. Store at -20°C for testing purposes. There are two types of urine testing, hemagglutination and hemostatic. The red blood cells used are Leghorn roosters, and the method is conventional.

Antiviral experiment: antiviral experiment was carried out in chicken embryos with different concentrations of dihydromyricetin Table 3 Influenza virus hemagglutination inhibition test serum dilution sample concentration stock solution 10× 20× 40× 80× 160× 320× 640× 1280× stock solution 1 ++++ ++++ ++++ ++++ ++++ ++++ ++ stock solution 2 ++++ ++++ ++++ ++++ ++++ ++++ ++ stock solution 3 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ stock solution 4 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 10 ^-5 a ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 10 ^-5b ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 10 ^-5 c ++++ ++++ ++++ ++++ ++++ ++++ ++ ++++ 10 ^-5 d ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 10 ^-6 a ++++ ++++ ++++ ++++ ++++ ++++ ++ ++++ 10 ^-6 b ++++ ++++ ++++ ++++ ++++ ++++ ++ ++++ 10 ^-6 c ++++ ++++ ++++ ++++ ++++ ++++ +++ ++ 10 ^-6 d ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ Remarks: 1. The original solution refers to the initial concentration of dihydromyricetin 4mg/ml

2. 10 ^-5 and 10 ^-6 refer to the dilution of the drug from the stock solution of dihydromyricetin

3. When judging the results, according to the hemagglutination intensity, respectively, ++++, +++, ++, - means that 50% of the red blood cells have agglutination, and the hemagglutination dilution degree of ++ is its hemagglutination decrease.

The red blood cells are fine sand-like and invalid at the bottom of the main hole (tube) is +++, that is, 100% agglutinated; the red blood cells are evenly spread on the bottom of the hole (tube), but the edges are not neat and slightly concentrated toward the bottom of the hole (tube) is ++ +, that is, 75% agglutination; the red blood cells form a ring at the bottom of the hole (tube), and those with small clots around it are ++, that is, 50% agglutination; the red blood cells form a round group at the bottom of the hole (tube), but the edge is not smooth enough, The one with a little clot around it is +, that is, 25% agglutination; the red blood cell forms a round group at the bottom of the hole (tube), and the one with smooth and neat edge is one, that is, no agglutination. Embodiment 8: in vitro antibacterial effect test: dihydromyricetin in vitro antibacterial effect test:

The antibacterial effect of the extracts was determined by plate punching method. After trying the test tube method and the plate punching method, the plate punching method, which is simple, easy and time-consuming, is determined. Five experimental strains were used: Staphylococcus aureus, Escherichia coli, pneumococcus, streptococcus hemolyticus, and catarrhal cocci in Table 4 for the effect test.

Experimental group strain name stock solution 1-fold dilution of the extract Polyclonal resistance (penicillin, streptomycin, nystatin) Staphylococcus aureus 9.5mm 8mm 17-20mm Escherichia coli 8.5mm 6mm 11mm pneumococcus - - 10mm Streptococcus hemolyticus - - 10-11mm Catharella - - 11mm Stock solution refers to the initial concentration of dihydromyricetin 4mg/ml Remarks: mm refers to the diameter of the inhibition zone

Embodiment 9:

1. Effect of dihydromyricetin on humoral immunity (serum lectin) in mice

1. Negative control group

2. Aspirin group

3. Dihydromyricetin large dose group of the present invention (0.15g/kg, maximum -1.35g/kg, dose group 0.45g/kg)

Observe the degree of agglutination, divide into five groups (0-4), calculate the antibody results for statistical processing, and calculate the volume of antibodies according to the formula (volume of antibodies Σ=S1+2S2+3S3+4S4...NSn)

Table 5, the influence of dihydromyricetin of the present invention on mouse lectin number of animals Anti-volume P value Blank control group 10 93.4±13.22 aspirin 10 115.00 ±17.31 <0.01 Dihydromyricetin small dose group 10 103±11.31 Middle dose group 10 115.92 ±19.38 <0.01 high dose group 10 103±7.01 <0.05 The results confirmed that: the middle and high dose groups significantly increased the anti-sheep hemagglutinin value in the serum, and there was a significant difference.

Example 10 Analgesic experiment of dihydromyricetin 1. Animal subject: Kunming mice 18-22 g male and female 2. Grouping and solution preparation:

The mice were divided into 5 groups with 10 mice in each group: ①Normal saline group; ②Water extraction group (0.26g/ml); ③Alcohol extract (0.21g/ml); ④Total flavonoids group (0.15g/ml) ; ⑤ dihydromyricetin (0.04g/ml). 3. Administration method

Each group of mice was weighed, administered by each 0.1ml/10g, administered by intragastric administration for three consecutive days, and opened 60min after administration for every 3 days, intraperitoneally fed 0.2ml/ of acetic acid (0.5%) solution, Observe the animal body count within 5-30 minutes. 4. Results and Statistics Drug Name Dosage Pain Relief Rate P Value Normal Saline 0.2ml/piece 0 Water Extraction Group 2.6g/kg 88.1% P＜0.05 Alcohol Extraction Group 2.1g/kg 93.5% P＜0.05 Total Flavonoids Group 1.05 /kg 88.0% P＜0.05 dihydromyricetin 0.4g/kg 54.1% P＜0.05 group results prove that dihydromyricetin has obvious analgesic effect.

Example 11 The lipid-lowering effect of dihydromyricetin on blood lipids in hyperlipidemic mice

Dose TC TG HDL-C group mg/kg (mmol/L) (mmol/ (mmol/L

                                                                                                               

0.78 0.37 0.28 Model control group 11.24± 2.11± 1.35±

                                                                                     

                                                       

1.90 0.2 0.56

The animals were randomly divided into 4 groups, normal control group, positive control group, model control group, and extract group. Ten mice in each group were given medicine once a day for 10 consecutive days, and the normal control group was not given any medicine. In fact, mice in each group were given 0.5ml/mouse of high-fat emulsion to form experimental hyperlipidemia. Fasted overnight after 10 days of medication. The next day, venous blood was collected from the orbits of the mice. Serum total cholesterol (TC), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) were detected by enzymatic method. The results showed that the serum TC and TG values of the model control group were significantly increased, and the HDL-C value was decreased. compared with the model control group. The dihydromyricetin group can significantly reduce serum TC and TG values, and also slightly increase HDL-C. It shows that the extract can inhibit the increase of blood lipid in mice caused by high-fat emulsion, and has a lipid-lowering effect.

Example 12 Dihydromyricetin to mice phenol red expectorant test experimental method:

Animals were orally administered test samples once a day for three consecutive days. The dose of the test product was 100/kg, the administration group was administered at 20ml/kg, and the negative control group was orally administered with the same volume of normal saline. Fasting 1 day before the last administration, intraperitoneal injection of 25% phenol red solution 500 mg/kg 0.5 hours after the last administration, and then 0.5 hours later, the animals were killed by neck, the trachea was separated, the injection needle was inserted, and 2ml of normal saline was used for flushing. 1M NaOH 0.1ml for color development, use a 722-type spectrophotometer to measure color at a wavelength of 546mm, and directly read the content of phenol red.

Bottom of the table: According to the test results, the dihydromyricetin sample increased the secretion of phenol red in Xiaozhi's respiratory tract after oral administration for 3 days, suggesting that the sample has an expectorant effect.

Table 1. Effect of dihydromyricetin on phenol red expectorant test in mice Sample Dose mg/kg Phenol red content increase percentage (ug/ml) (X±SD) ％ Dihydromyricetin 20ml/kg 0.68±0.22 -Dihydromyricetin 100 0.70± -0.17* *P>0.05, **P<0.05, ***P<0.01 compared with normal saline group.

Claims (9)

1. the pharmaceutical composition that contains dihydromyricetin and pharmaceutical acceptable carrier and excipient.

2. the purposes of dihydromyricetin in the medicine of preparation treatment hepatitis and/or hepatoprotective.

3. dihydromyricetin is in the purposes of preparation in the antibacterial-anti-inflammatory drug.

4. dihydromyricetin is in the medicine of preparation enhance immunity systemic immunity power and the purposes in the health product.

5. dihydromyricetin is in preparation analgesia, blood fat reducing, the purposes in the expectorant medicine.

6. the compositions of claim 1, it is a tablet.

7. the compositions of claim 1, it is an electuary.

8. the compositions of claim 1, it is a capsule.

9. the compositions of claim 1, it is a syrup.