CN1596888A - Application of dihydromyricetrin - Google Patents

Abstract

An application of dihydromyricetin in preparing the antalgic and apophlegmatic medicine or health-care food is disclosed.

Description

The new purposes of dihydromyricetin

The invention belongs to the health care of treatment hepatitis, hepatoprotective, analgesia, blood fat reducing is eliminated the phlegm and the natural medicine field of anti-inflammation.

Document Walter Karrerr, Birkhauser Verlag, Bassel undstuttgart (1958), P.652, NO:1640 discloses the structural formula of dihydromyricetin,

But the activity of dihydromyricetin is not studied.

The applicant is surprisingly found out that dihydromyricetin has treatment hepatitis, the hepatoprotective health care, and anti-inflammation, analgesia, blood fat reducing is eliminated the phlegm and the effect of enhance immunity systemic immunity power.Dihydromyricetin can be made pharmaceutical composition with pharmaceutical acceptable carrier or mixed with excipients.Dihydromyricetin can obtain from vitaceae.Also can obtain by the chemical field conventional method.

An object of the present invention is to provide a kind of pharmaceutical composition that contains dihydromyricetin and pharmaceutical acceptable carrier or excipient for the treatment of hepatitis.

Another object of the present invention provides the pharmaceutical composition that contains dihydromyricetin and pharmaceutical acceptable carrier or excipient that contains anti-inflammation.

A further object of the present invention provides a kind of pharmaceutical composition that contains dihydromyricetin and health product of hepatoprotective health care.

Another object of the present invention provides the pharmaceutical composition that contains dihydromyricetin and the health product of enhance immunity systemic immunity power.

Another object of the present invention provides the pharmaceutical composition that contains dihydromyricetin or the medicine for external use of anti-inflammatory analgesic.

Another object of the present invention provides the pharmaceutical composition that contains dihydromyricetin and the health product of blood fat reducing.

Another object of the present invention provides pharmaceutical composition and the health product that expectorant contains dihydromyricetin.

Another object of the present invention is a dihydromyricetin in preparation treatment hepatitis, hepatoprotective effect, and antiinflammatory, analgesia, blood fat reducing is eliminated the phlegm or is improved purposes in the pharmaceutical composition of immunity.

Dihydromyricetin can obtain like this: decoct vitaceae by all kinds of solvents (water and/or organic solvent), decoct one or many, decoction liquor is through concentrating cooling, standing over night, detection precipitates the active ingredient with supernatant, gets to have active precipitation, obtains crystal after chromatographic isolation

Principle of the present invention is to follow the trail of the dihydromyricetin drug activity by setting up the pharmacological screening model, finds its pharmaceutical usage.The animal model test of the influence of the test of the influence of the mouse experiment liver damage that carbon tetrachloride is caused and mouse experiment liver damage that D-Gal is caused shows that dihydromyricetin has tangible hepatoprotective effect; Extracorporeal bacteria inhibitor test and and antivirus test show that dihydromyricetin has antibacterial and anti-inflammation functions; The test of serum agglutinin shows that dihydromyricetin has the effect of enhance immunity systemic immunity power.

Find that dihydromyricetin can be used for pharmaceutical field, field of health care products and field of food.

Dihydromyricetin can be directly medicinal or can be mixed with pharmaceutical composition with pharmaceutically acceptable carrier or mixed with excipients, and dihydromyricetin can also be as the effective ingredient of health product, and the present composition can also be mixed with beverage etc.

Pharmaceutical composition of the present invention can be mixed with pharmaceutical composition by conventional preparation technique of drug world and carrier.The present composition can be prepared into tablet, granule, and capsule, medicine for external use can also join in tea or the beverage, makes health tea or beverage.

The regular convention formula of tablet can be a dihydromyricetin: the 10%-90% weight ratio, and lactose 90%-10% weight ratio, sodium carboxymethyl cellulose 1.5g, magnesium stearate 0.5g, 50% ethanol is an amount of.The regular convention formula of electuary can be: dihydromyricetin 5-100% weight ratio, (sucrose+dextrin) 95-0% weight ratio, sucrose: dextrin=2: 1.Capsular regular convention formula can be: dihydromyricetin 10-100% weight ratio, starch 90-0% weight ratio.But those skilled in the art also can be mixed with other dosage form with technology well known in the art and carrier as required.

Describe the present invention in detail by the following examples.But should be appreciated that these embodiment just illustrate the present invention, rather than in office where face limits the scope of the invention.

Embodiment 1: tablet:

Dihydromyricetin 25g,

Lactose 53g,

Sodium carboxymethyl cellulose 1.5g,

Magnesium stearate 0.5g,

50% ethanol 5ml

Said components is mixed the back compacting in flakes in mixer.

Embodiment 2: syrup

1 part of weight of dihydromyricetin,

2.5 parts of weight of sucrose,

1.25 parts of weight of dextrin,

5 parts of weight of ethanol

Mix mentioned component and obtain syrup.

Embodiment 3: electuary

Dihydromyricetin 115g,

Icing Sugar 345g,

Dextrin 145g,

Ethanol 5ml

Said components is mixed after drying, obtains electuary.

Embodiment 4: capsule

Dihydromyricetin 100g,

Starch 20g,

Behind the said components mix homogeneously, incapsulate.

Embodiment 5: the influence of the mouse test liver damage that dihydromyricetin causes carbon tetrachloride:

Animal is divided into six groups at random: three groups of normal control groups, positive controls, model control group, dihydromyricetin group: 100mg/kg, 200mg/kg, 400mg/kg.The normal control group is that animal is not given any medicine, only gives solvent; Positive controls is bifendate 200mg/kg, qdx6, and po, bifendate is treating hepatitis medicine commonly used, with bifendate treatment animal pattern, observe the curative effect, in contrast; Model control group is the check drug effect, and animal must be made hepatitis model earlier, makes animal suffer from hepatitis; The dihydromyricetin group is on above-mentioned model basis, gives dihydromyricetin again, and with observe the curative effect, three groups all is Bidx6 po.Each treated animal grouping administration, every group of 10 mices, twice on the one, take volume 0.5ml/20g mice, took continuously six days, 0.2ml/ mice of 1 hour lumbar injection 1% carbon tetrachloride oil solution after the last administration, on an empty stomach after 16 hours by the eye socket venous blood collection, separation of serum, the SGPT (ALT) of mensuration serum, SGOT (AST) content and serum total bilirubin (T-BIL caffeine sodium benzoate colorimetry) content, and and positive controls, the normal control group, model control group relatively, model control group is taken the equivalent coordinative solvent, positive controls adopts bifendate 200mg/kg, pox6, once a day.The results are shown in Table 1.

Table 1. dihydromyricetin causes the variation of the serum SGOT of mouse liver injury to carbon tetrachloride

Group	Dosage mg/kg	Serum SGPT (U) X ± SD	Serum SGOT (U) X ± SD	?T-BIL
						Coordinative solvent
Model control group	Coordinative 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

* compare with model group P＜0.05 * * P＜0.01

From the effect of dihydromyricetin with excellent treatment hepatitis as can be seen of the data of table 1.And reduce T-BIL content, show the hepatoprotective effect of dihydromyricetin with tangible reducing enzyme and treating jaundice.

Embodiment 6: dihydromyricetin causes the protective effect of mouse liver injury to D-galactosamine

Animal is divided into six groups at random, and positive controls, model control group, normal control group, dihydromyricetin component are three groups: 100mg/kg, and 200mg/kg, 400mg/kg, the meaning of each group is with above-mentioned embodiment 4.Normal control group, model control group are taken corresponding solvent, positive controls lumbar injection 200mg/kg, pox6, qd, 1 hour D-galactosamine 100mg/kg gets blood by eye socket after 16 hours, separation of serum after the last administration, measure serum SGPT, the variation of serum SGOT the results are shown in Table 2

Table 2 dihydromyricetin causes the serum SGPT of mouse liver injury, the variation of SGOT to D-galactosamine

Group	Dosage mg/kg	Serum SGPT (U) X Xu D	Serum SGOT (U) X Xu D
				The normal control group	Coordinative solvent	＜40	33.7_4.7
Model control group	Coordinative 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, * * P＜0.01 and model group comparison

From the data of table 2 as can be seen dihydromyricetin have fabulous hepatoprotective effect.

Embodiment 7: dihydromyricetin is tested at external antibacterial, antiviral:

Separation of influenza virus and mirror (dripping) are fixed

Embryo Gallus domesticus at present commonly used separates and carries out influenza virus, and route of inoculation is the allantoic cavity of Embryo Gallus domesticus, occurs the characteristics of lesion of back chicken embryo death after 3 days and hemagglutinin occurs, and to this, we adopt the method to carry out evaluation to dihydromyricetin.We adopt the influenza virus of certain density extracting solution and 4 HAUs to mix at 1: 1, allow the two inject Embryo Gallus domesticus after 20 minutes again in the room temperature effect, and injection volume is every embryo 0.2ml.The Embryo Gallus domesticus of 9 to 11 ages in days is adopted in experiment, carry out the allantoic cavity inoculation, cultivate after 48 hours, placed liquid in 4 ℃, aseptic then its urine of getting, get supernatant through 1000rpm after centrifugal 10 minutes, packing is deposited for-20 ℃, the usefulness that is equipped with detection, the detection of urine divides blood clotting and blood to press down two kinds, and the erythrocyte of use comes prosperous cock, and method is a conventional method.

The antiviral experiment: the dihydromyricetin with variable concentrations carries out the antiviral experiment in Embryo Gallus domesticus

Table 3 hirst's hemagglutination inhibition test

The serum dilution sample concentration	Stock solution	10 ×	20 ×	40 ×	80 ×	160 ×	320 ×	640 ×	128 0×
										Stock solution 1	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	++
Stock solution 2	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	++
										Stock solution 3	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	+++ +	++
Stock solution 4	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	+++ +	++
										10 -5a	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	+++ +	+++ +	++
10 -5b	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	+++ +	+++ +	++
										10 -5c	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	++	+++ +
10 -5d	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	+++ +	++
										10 -6a	+++ +	++ ++	++ ++	+++ +	++ ++	+++ +	++	+++ +

10 -6b	+++ +	??++ ??++	??++ ??++	??+++ ??+	??++ ??++	??+++ ??+	??++	??+++ ??+
										10 -6c	+++ +	??++ ??++	??++ ??++	??+++ ??+	??++ ??++	??+++ ??+	??+++ ??+	??++
10 -6d	+++ +	??++ ??++	??++ ??++	??+++ ??+	??++ ??++	??+++ ??+	??+++ ??+	??++

Remarks: 1, stock solution refers to the dihydromyricetin 4mg/ml of initial concentration

2,10 ^-5With 10 ^-6Finger carries out the dilution factor of medicine from dihydromyricetin stock solution

3, during result of determination by blood clotting intensity respectively with ++ ++, +++, ++ ,-expression coagulation appears with 50% erythrocyte ++ the blood clotting dilution factor be its blood clotting decline

Erythrocyte is the invalid main aperture of fine sand grain sample (pipe) end person +++, i.e. 100% coagulation; Erythrocyte evenly is laid on the end, hole (pipe), but the edge is whole to hole (pipe) end concentrator is not slightly +++, i.e. 75% coagulation; (pipe) end, form a ring-type to erythrocyte in the hole, has little coagulation piece person to be all around ++, i.e. 50% coagulation; (pipe) end, form wad to erythrocyte in the hole, but the edge is smooth inadequately, have slightly all around coagulation piece person for+, i.e. 25% coagulation; (pipe) end, form wad to erythrocyte in the hole, and the smooth neat person in edge is one, does not promptly have coagulation.

Embodiment 8: external bacteriostasis test:

The external inhibitory effect test of dihydromyricetin:

Adopt dull and stereotyped punch method to measure the antimicrobial effect of extract.After test tube method and dull and stereotyped punch method are attempted, determined this short method simple and easy to do and consuming time of dull and stereotyped punch method.Adopt five kinds of experimental strains: staphylococcus aureus, escherichia coli, streptococcus pneumoniae, hemolytic chain bacterium,

Table 4 micrococcus catarrhalis carries out effect test.

The experimental group strain name	Stock solution	1 times of dilution of extracting solution	Multi-resistance (penicillin and streptomycin, nystatin)
				Staphylococcus aureus	????9.5mm	?8mm	?17-20mm
Escherichia coli	????8.5mm	?6mm	?11mm

Streptococcus pneumoniae	-	-	????10mm
				The hemolytic chain bacterium	-	-	????10-11mm
The mucositis fungus ball	-	-	????11mm

Stock solution refers to the dihydromyricetin 4mg/ml of initial concentration

Remarks: mm refers to the diameter of inhibition zone

Embodiment 9:

Dihydromyricetin influences mouse humoral immune (serum agglutinin)

1, negative control group

2, aspirin group

3, dihydromyricetin of the present invention is heavy dose of organizes (0.15g/kg, big-1.35g/kg, dosage group 0.45g/kg)

Observe the coagulation degree, divide five groups (0-4), calculating antibody result carries out statistical procedures, is calculated as follows the antibody product

(antibody product ∑=S1+2S2+3S3+4S4 ... NSn)

Table 5, dihydromyricetin of the present invention are to the influence of mice agglutinin

	Number of animals	The antibody product	The P value
				The blank group	10	93.4±13.22
Aspirin	10	115.00± 17.31	＜0.01
				The dihydromyricetin small dose group	10	103±11.31
Middle dosage group	10	115.92± 19.38	＜0.01
				Heavy dose of group	10	103±7.01	＜0.05

The result confirms: middle and high dosage group obviously increases anti-sheep red blood cell agglutinin value in the serum, and significant difference is arranged, and the dihydromyricetin higher dosage has potentiation to system's phagocytic function in the body dictyosome.

Embodiment 10

The dihydromyricetin analgesic experiment

1, animal target: Kunming white mice 18～22g male and female half and half

2, grouping and solution preparation:

With 10 every group of white mice, be divided into 5 groups: 1. normal saline group; 2. water extraction group (0.26g/ml); 3. alcohol extract (0.21g/ml); 4. total flavones group (0.15g/ml); 5. dihydromyricetin (0.04g/ml).

3, administering mode

Weigh to each group white mice, by every 0.1ml/10g administration, continuous three days gastric infusions, and open 60min after the administration in per 3 days, the abdominal cavity only feeds acetic acid (0.5%) solution 0.2ml/, observes in 5～30min animal and sweeps the body number.

4, result and statistics

Medicine name dosage pain relieving rate P value

Normal saline 0.2ml/ only 0

Water is carried group 2.6g/kg 88.1% P＜0.05

Alcohol extraction group 2.1g/kg 93.5% P＜0.05

Total flavones group 1.5g/kg 88.0% P＜0.05

Dihydromyricetin 0.4g/kg 54.1% P＜0.05

Group

The result proves: dihydromyricetin has tangible analgesic effect.

Embodiment 11

Dihydromyricetin is to the effect for reducing fat of hyperlipidemia mice blood fat

Dosage TC TG HDL-C

Group mg/kg (mmol/L (mmol/ (mmol/L

)??????????L)????????)

Normal control group 3.91 ± 1.41 ± 1.85 ±

0.78???????0.37??????0.28

Model control group 11.24 ± 2.11 ± 1.35 ±

5.79???????0.41??????0.52

Clofibrate 310 5.12 ± 1.76 ± 1.72 ±

1.14???????0.11??????0.64

Dihydromyricetin 400 6.11 ± 1.58 ± 1.621 ±

1.90???????0.2???????0.56

Animal is divided into 4 groups at random, normal control group, positive controls, model control group, extracting solution group.Every group of 10 mices, once-a-day, successive administration 10 days, the normal control group is not given any medicine.All only respectively organize mice in fact, form experimental hyperlipidemia to high lipoprotein emulsion 0.5ml/.Overnight fasting after medication in 10 days.Next day is from the mouse orbit extracting vein blood.Press enzyme process and detect serum total cholesterol (TC), triglyceride (TG), HDL-C (HDL-C) content.The result shows, the model control group serum TC, and the TG value obviously raises, and the HDL-C value descends.Compare with model control group.The dihydromyricetin group can obviously reduce serum TC, and the TG value also makes HDL-C slightly raise.Illustrate that this extract can suppress the mice blood fat rising that high lipoprotein emulsion causes, has effect for reducing fat.

Embodiment 12

Dihydromyricetin is to the phenol red expectorant test of mice

Experimental technique:

Animal oral test every day sample 1 time, continuous oral three days.Being subjected to test product dosage is 100/kg, and the administration group is pressed the 20ml/kg administration, and negative control group is oral with the volume normal saline.Last administration fasting in preceding 1 day, 0.5 hour lumbar injection 25% phenol red solution 500mg/kg after the last administration, press neck again after 0.5 hour and put to death animal separation trachea, insert entry needle, use the 2ml normal saline flushing, flushing liquor adds 1M NaOH 0.1ml colour developing, in 546mm wavelength colorimetric, directly reads phenol red content with 722 type spectrophotometers.

The table below: represent that from result of the test the dihydromyricetin sample increases the phenol red secretory volume of little will respiratory tract after oral 3 days, the prompting sample has phlegm-dispelling functions.

Table 1, dihydromyricetin are to the influence of the phenol red expectorant test of mice

Sample dose mg/kg	Phenol red amount (increasing percentage rate ug/ml)
		(X±SD)?????％
	Dihydromyricetin 20ml/kg dihydromyricetin 100		0.68±0.22??- 0.70±??????- 0.17*

* P＞0.05, * * P＜0.05, * * * P＜0.01 are with physiology saline group relatively.

Claims (2)

1. the purposes of dihydromyricetin in preparation analgesia, expectorant medicine.

2. the purposes of dihydromyricetin in preparation analgesia, expectorant health product.