CN113768896A

CN113768896A - Gardenia duodenum positioning preparation for treating cholestatic liver disease

Info

Publication number: CN113768896A
Application number: CN202111268224.7A
Authority: CN
Inventors: 梁爱华; 王锦玉; 田婧卓; 赵雍; 易艳; 仝燕; 李春英
Original assignee: Institute of Materia Medica of CAMS
Current assignee: Institute of Materia Medica of CAMS
Priority date: 2021-10-28
Filing date: 2021-10-28
Publication date: 2021-12-10
Anticipated expiration: 2041-10-28
Also published as: CN113768896B

Abstract

The invention relates to the field of traditional Chinese medicine preparations, in particular to a preparation method of a gardenia extract and a gardenia duodenum positioning preparation for treating cholestatic liver diseases. The preparation method of fructus Gardeniae extract comprises decocting fructus Gardeniae in water, passing the extractive solution through SP825L macroporous resin column at a speed of 2-4BV/h, collecting eluate, concentrating, and drying. The gardenia duodenal positioning preparation comprises a tablet core and an enteric coating wrapping the tablet core; the tablet core comprises fructus Gardeniae extract, lactose, microcrystalline cellulose and magnesium stearate; the enteric coating comprises No. I and No. II acrylic resin and diethyl phthalate. The active ingredients extracted by the preparation method are high in content, and the prepared gardenia duodenal positioning preparation is moderate in hardness, strong in moisture resistance, capable of rapidly disintegrating in duodenal fluid and being absorbed by organisms, and suitable for clinical emergencies such as cholestatic hepatitis and the like.

Description

Gardenia duodenum positioning preparation for treating cholestatic liver disease

Technical Field

The invention relates to the field of traditional Chinese medicine preparations, in particular to a preparation method of a gardenia extract and a gardenia duodenum positioning preparation for treating cholestatic liver diseases.

Background

Cholestasis refers to the pathological condition of bile formation, secretion and excretion disorder caused by various reasons inside and outside the liver, and the bile cannot normally flow into the duodenum and enter the blood, and clinically, the symptoms can be manifested as pruritus, hypodynamia, dark urine color, jaundice and the like. Early patients mainly show that serum ALP and GGT levels are increased, hyperbilirubinemia can appear after the disease condition is aggravated, and liver fibrosis, cirrhosis and even liver cancer can be caused if the treatment is not carried out in time.

The drugs for treating cholestasis are limited, ursodeoxycholic acid is the only drug which is currently approved in the United states and widely used for treating cholestasis, is hydrophilic dihydroxycholic acid, is separated from Chinese black bear bile at the earliest, exists in a small amount (about 1-3 percent of the whole bile acid pool) as secondary cholic acid in a human body, and is 7 b-isomer formed by chenodeoxycholic acid under the action of colonic bacteria. Oral administration enhances absorption of bile acids by dissolution. In addition, phenobarbital can enhance bile flow, bile acid excretion, and liver blood flow. Glucocorticoids can relieve inflammation of capillary bile ducts and improve bile flow, and are common medicines for treating cholestatic liver diseases. However, the chemical drugs mainly aim at the treatment of the etiology and the related complications, and because the pathogenesis of the disease is complex, no specific treatment drug is available at present, and the patients are seriously troubled by the accompanied systemic pruritus and the expensive medical cost.

The disease without cholestasis in the traditional Chinese medicine belongs to the category of jaundice in the traditional Chinese medicine according to the clinical manifestations of yellow body and eye, pruritus of whole body, yellow and red urine and grey and white stool. The traditional Chinese medicine treatment has many attempts, the pathogenesis of damp-heat evil is further deeply researched, the traditional heat clearing and dampness eliminating treatment principle and treatment method are broken through, diversified and multi-path administration is gradually carried out, the curative effect is better, the side effect is small, and the advantages and the characteristics of the traditional Chinese medicine are shown.

The Chinese patent application CN105944032A discloses a traditional Chinese medicine composition for treating intrahepatic cholestatic jaundice and an application thereof, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight: 9 parts of radix bupleuri, 30 parts of oriental wormwood, 9-15 parts of cape jasmine fruit, 9-15 parts of prepared rhubarb, 30 parts of red sage root, 15 parts of radix curcumae, 30 parts of plantain herb, 60-90 parts of red paeony root, 30-45 parts of bighead atractylodes rhizome, 30-60 parts of barbed skullcap herb, 60 parts of lalang grass rhizome and 9 parts of raw liquorice. The prescription has more medicines, has certain curative effect on intrahepatic cholestatic jaundice, has pending further improvement on the utilization ratio and the efficacy of medicinal materials, is not suitable for being prepared into Chinese patent medicines, and has poor applicability especially to patients with jaundice clinical emergency.

The Chinese patent application CN111138557A discloses a gardenia polysaccharide and a preparation method and application thereof, wherein the gardenia polysaccharide comprises the following raw materials in percentage by mass: the total sugar content is 41-56%, uronic acid content is 34-45%, and protein content is 7-10%. The preparation method of the gardenia polysaccharide comprises the steps of crushing gardenia medicinal materials, removing fat, adding water, extracting, precipitating by ethanol, repeatedly freezing and thawing for protein removal, dialyzing, and freeze-drying. The invention researches the extraction and curative effect of gardenia polysaccharide, but the extraction and the medicinal effect of a large amount of active ingredients except saccharide in gardenia are needed to be researched and developed, and the medicinal material resources are more fully utilized.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides a method for extracting a gardenia extract and a gardenia duodenum positioning preparation for treating cholestatic liver diseases. The fructus Gardeniae duodenal locating preparation has high content of active ingredients, moderate hardness, strong moisture resistance, no splintering phenomenon in artificial gastric juice within 120min, rapid disintegration in artificial duodenal juice, rapid disintegration after entering intestinal tract, and absorption by organism, and quick action, and is suitable for clinical emergency such as jaundice, cholestatic hepatitis, etc.

The purpose of the invention is realized by the following technical scheme:

a method for preparing Gardenia jasminoides Ellis extract for treating cholestatic liver disease comprises the following steps:

(1) decocting fructus Gardeniae in water to obtain decoction;

(2) passing the water decoction through SP825L macroporous resin column at a speed of 2-4BV/h, washing with water, eluting with ethanol, collecting ethanol eluate, concentrating, and drying.

Preferably, the mass ratio of the volume of the added water to the gardenia in the step (1) is 10-15:1 mL/g; the decoction is carried out for 2 times, and the decoction time is 0.5-2 h.

Preferably, the water washing in the step (2) is water washing with 1-3BV, and the ethanol elution is elution with 3-5BV of 30-70% ethanol; preferably 30% ethanol.

Preferably, the diameter-height ratio of the SP825L macroporous resin column in the step (2) is 1:1.3-1:9.2, and the mass ratio of the crude drug in the water decoction to the SP825L macroporous resin column is 1: 1-3.

Preferably, the concentration in the step (2) is to concentrate the alcohol eluent to an extract with the relative density of 1.21-1.25 at 60 ℃; the drying is spray drying or vacuum drying.

Preferably, the inlet temperature of the spray drying is 85-190 ℃, and the outlet temperature is fixed at 75-100 ℃;

preferably, the vacuum drying is 60-80 ℃;

preferably, the vacuum drying is 70 ℃.

Preferably, the content of geniposide, genipin gentiobioside and hydroxy-isogeniposide in the obtained gardenia extract is respectively 400-750mg/g, 90-150mg/g and 10-50 mg/g.

The invention also aims to provide a gardenia duodenal positioning preparation which comprises a tablet core and an enteric coating wrapping the tablet core.

Preferably, the tablet core comprises the following components in parts by weight: 35-45 parts of gardenia extract, 50-70 parts of lactose, 15-25 parts of microcrystalline cellulose and 0.15-0.35 part of magnesium stearate;

preferably, the enteric coating comprises acrylic resin No. I, acrylic resin No. II and diethyl phthalate.

Preferably, the mass ratio of lactose to microcrystalline cellulose is 2-4: 1.

Preferably, the mass ratio of the No. I acrylic resin to the No. II acrylic resin is 1: 4.5-5.5.

Preferably, the mass of the enteric coating is 8-12% of the mass of the tablet core;

preferably, the ratio of the mass of the diethyl phthalate to the total mass of the No. I acrylic resin latex and the No. II acrylic resin is 0.15-0.25: 1.

the invention also aims to provide application of the gardenia extract obtained by the preparation method or the gardenia duodenum positioning preparation in preparing a medicament for treating cholestatic liver diseases.

Compared with the prior art, the invention has the beneficial effects that:

(1) the active ingredients extracted by the extraction method are high in content, the prepared gardenia duodenum positioning preparation is moderate in hardness and strong in moisture resistance, does not crack in artificial gastric juice within 120min, can be rapidly disintegrated in artificial duodenal juice, can be rapidly disintegrated after entering intestinal tracts and absorbed by organisms, has quick-acting effect, and is suitable for clinical emergencies such as jaundice and cholestatic hepatitis.

(2) The invention adopts a specific resin column for separation and purification, obviously improves the content of active ingredients, and the known effective ingredient proportion is 60-95 percent, which is better in the aspect of treating cholestatic hepatitis;

(3) according to the invention, through research on enteric coating materials, the specific acrylic resin polymer compound is found to achieve a better oriented duodenal disintegration effect.

Detailed Description

The present invention will be further described with reference to the following embodiments.

Example 1

(1) Preparing a gardenia extract:

1250g of gardenia medicinal material is taken, 15 times of water is added for decoction for the first time, 10 times of water is added for decoction for the second time, the water decoctions are combined, the mixture is cooled to room temperature and filtered, and processed SP825L resin is taken to be packed into a column by a wet method, the diameter-height ratio is 1:2.7, and the weight ratio of the raw materials is as follows: the resin amount is 1:2 sample application of the gardenia liquid medicine, 3BV/h sample application, 2BV distilled water washing for removing impurities, 4BV 30% ethanol elution, collection of alcoholysis liquid absorption, reduced pressure recovery of ethanol to an extract with a relative density of 1.21(60 ℃), and vacuum drying at 70 ℃ to obtain the gardenia liquid medicine. In the obtained extract, the content of geniposide is 742mg/g, the content of genipin gentiobioside is 150mg/g, and the content of hydroxygardenia glycoside is 48 mg/g.

(2) Gardenia duodenum positioning preparation

Tablet core prescription: 40 parts of gardenia extract, 20 parts of microcrystalline cellulose, 60 parts of lactose and 0.25 part of magnesium stearate

Preparation of the tablet core: mixing fructus Gardeniae extract, microcrystalline cellulose, lactose, and magnesium stearate 0.125 parts, granulating by dry method, adding magnesium stearate 0.125 parts, mixing, and tabletting to obtain tablet core;

the prescription of the enteric layer: the tablet comprises No. I acrylic resin, No. II acrylic resin and diethyl phthalate, wherein the mass of an enteric coating layer is 8% of that of a tablet core, the mass ratio of the No. I acrylic resin latex to the No. II acrylic resin is 1:5, and the mass ratio of the diethyl phthalate to the sum of the mass of the No. I acrylic resin latex and the No. II acrylic resin is 0.2: 1.

Preparing enteric-coated layer coating liquid: dissolving polyacrylic resin I and polyacrylic resin II in 95% ethanol according to the mass ratio of 1:1, and uniformly stirring to obtain a phase A; adding plasticizer diethyl phthalate into 95% ethanol at a mass ratio of 1:0.5, and homogenizing with a high shear homogenizer for 10min to obtain phase B. Slowly pouring the suspension of the B into the A, and sieving by a 100-mesh sieve to obtain the compound.

The enteric coating step is as follows:

coating with rolling method at a coating pan inclination angle of 45 deg. and rotation speed of (30 + -1) r.min^-1The air inlet temperature is (40 +/-1) DEG C, the atomization pressure is 0.4kg cm^-2The flow rate of the spray liquid is 5.5 ml/min^-1The concentration of the coating solution was 6.25% (g/100 mL). Curing for 10 hours to obtain the product.

Example 2

(1) Preparing a gardenia extract:

taking a gardenia medicinal material, adding 12 times of water for decocting for 1 hour for the first time, adding 12 times of water for decocting for 2 hours for the second time, combining the water decoctions, cooling to room temperature, filtering by using non-woven fabrics, taking the treated SP825L resin, packing into a column by a wet method, wherein the diameter is 1:9.2, and the weight is calculated as the amount of the crude drugs: loading fructus Gardeniae liquid at a resin amount of 1:1, loading 2BV/h, washing with 1BV distilled water to remove impurities, eluting with 3BV 70% ethanol, collecting alcoholysis solution, recovering ethanol under reduced pressure to obtain extract with relative density of 1.25(60 deg.C), and vacuum drying at 60 deg.C. The obtained extract contains geniposide 435mg/g, genipin gentiobioside 140mg/g, and hydroxygeniposide 39mg/g

(2) Gardenia duodenum positioning preparation

Tablet core prescription: 35 parts of gardenia extract, 15 parts of microcrystalline cellulose, 50 parts of lactose and 0.15 part of magnesium stearate

Preparation of the tablet core: mixing fructus Gardeniae extract, microcrystalline cellulose, lactose, and magnesium stearate 0.075 part, granulating by dry method, adding magnesium stearate 0.075 part, and tabletting to obtain tablet core;

the prescription of the enteric layer: the tablet comprises No. I acrylic resin, No. II acrylic resin and diethyl phthalate, wherein the mass of an enteric-coated layer is 10% of that of a tablet core, the mass ratio of No. I acrylic resin latex to No. II acrylic resin is 1:4.5, and the ratio of diethyl phthalate to (the sum of the mass of the No. I acrylic resin latex and the mass of the No. II acrylic resin) is 0.15: 1.

Preparing enteric-coated layer coating liquid: dissolving polyacrylic resin I and polyacrylic resin II in 95% ethanol according to the mass ratio of 1:0.5, and uniformly stirring to obtain a phase A; adding plasticizer diethyl phthalate into 95% ethanol at a mass ratio of 1:0.5, and homogenizing with a high shear homogenizer for 10min to obtain phase B. Slowly pouring the suspension of the B into the A, and sieving by a 100-mesh sieve to obtain the compound.

The enteric coating step is as follows:

coating with rolling method at a coating pan inclination angle of 45 deg. and rotation speed of (30 + -1) r.min^-1The air inlet temperature is (40 +/-1) DEG C, the atomization pressure is 0.4kg cm^-2The flow rate of the spray liquid is set to be 5 ml/min^-1The concentration of the coating solution is 6% (g/100 mL). Curing for 10 hours to obtain the product.

Example 3

(1) Preparing a gardenia extract:

taking a gardenia medicinal material, adding 15 times of water for decocting for 2 hours for the first time, adding 12 times of water for decocting for 1 hour for the second time, combining water decoctions, cooling to room temperature, filtering by using non-woven fabrics, taking processed SP825L resin, packing into a column by a wet method, wherein the diameter-height ratio is 1:1.3, and taking the crude drugs: loading fructus Gardeniae liquid at a resin amount of 1:3, loading at a rate of 4BV/h, washing with 3BV distilled water to remove impurities, eluting with 5BV 50% ethanol, collecting alcoholysis solution, recovering ethanol under reduced pressure to obtain extract with relative density of 1.25(60 deg.C), spray drying at inlet temperature of 85 deg.C and outlet temperature of 75 deg.C. In the obtained extract, the geniposide content is 627mg/g, the genipin gentiobioside content is 96mg/g, and the hydroxygardenia glycoside content is 17 mg/g;

(2) gardenia duodenum positioning preparation

Tablet core prescription: 45 parts of gardenia extract, 25 parts of microcrystalline cellulose, 70 parts of lactose and 0.35 part of magnesium stearate

Preparation of the tablet core: mixing fructus Gardeniae extract, microcrystalline cellulose, lactose, and magnesium stearate 0.175 parts, granulating by dry method, adding magnesium stearate 0.175 parts, and tabletting to obtain tablet core;

the prescription of the enteric layer: the tablet comprises a No. I acrylic resin latex, a No. II acrylic resin and diethyl phthalate, wherein the mass of an enteric coating layer is 12% of that of a tablet core, the mass ratio of the No. I acrylic resin latex to the No. II acrylic resin is 1:5.5, and the ratio of the diethyl phthalate to (the sum of the mass of the No. I acrylic resin latex and the mass of the No. II acrylic resin) is 0.25: 1.

Preparing enteric-coated layer coating liquid: dissolving polyacrylic resin I and polyacrylic resin II in 95% ethanol according to the mass ratio of 1:0.6, and uniformly stirring to obtain a phase A; adding plasticizer diethyl phthalate into 95% ethanol at a mass ratio of 1:0.6, and homogenizing with a high shear homogenizer for 10min to obtain phase B. Slowly pouring the suspension of the B into the A, and sieving by a 100-mesh sieve to obtain the compound.

The enteric coating step is as follows:

coating with rolling method at a coating pan inclination angle of 45 deg. and rotation speed of (30 + -2) r.min^-1The air inlet temperature is (40 +/-2) DEG C, the atomization pressure is 0.4kg cm^-2The flow rate of the spray liquid is 6 ml/min^-1The concentration of the coating solution is 5% (g/100 mL). Curing for 10 hours to obtain the product.

Comparative example 1

The difference between the comparative example and the example 1 is that HPD100 is adopted in the column in the step (1), the column is soaked for 24 hours by adding 95% ethanol before the column is used, distilled water is eluted to be neutral, and the rest is consistent with the example 1.

Comparative example 2

The difference between the comparative example and the example 1 is that HPD600 is adopted in the column in the step (1), the column is soaked for 24 hours by adding 95% ethanol before being used, distilled water is eluted to be neutral, and the rest is kept consistent with the example 1.

Comparative example 3

The difference between the comparative example and the example 1 is that NKAII is used as the column in the step (1), the NKAII is soaked in 95% ethanol for 24h, and distilled water is eluted to neutrality, and the rest is the same as the example 1.

Comparative example 4

The difference between the comparative example and the example 1 is that HPD400 is used as the column in the step (1), the column is soaked in 95% ethanol for 24h, and distilled water is eluted to be neutral, and the rest is consistent with the example 1.

Comparative example 5

The difference between the comparative example and the example 1 is that the diameter-height ratio of the wet column packing in the step (1) is 1: 12, the rest remaining in accordance with example 1. The obtained extract contains geniposide 416mg/g, genipin gentiobioside 125mg/g, and hydroxygeniposide 12 mg/g.

Comparative example 6

The difference between this comparative example and example 1 is that the lactose, the prescription raw material for the tablet core in step (2), is replaced by a compressible starch, the rest remaining the same as in example 1.

Comparative example 7

The difference between the comparative example and the example 1 is that the mixture ratio of the raw materials of the tablet core in the step (2) is different, specifically 40 parts of gardenia extract, 10 parts of microcrystalline cellulose, 70 parts of lactose and 0.25 part of magnesium stearate, and the rest is consistent with the example 1.

Comparative example 8

The difference between the comparative example and the example 1 is that the mixture ratio of the raw materials of the tablet core in the step (2) is different, specifically 40 parts of gardenia extract, 40 parts of microcrystalline cellulose, 40 parts of lactose and 0.2 part of magnesium stearate, and the rest is consistent with the example 1.

Comparative example 9

The difference between this comparative example and example 1 is that the prescribed lactose material for the tablet core in step (2) is replaced by sodium carboxymethyl starch, the rest remaining the same as in example 1.

Comparative example 10

The difference between the comparative example and the example 1 is that the mass ratio of the No. I acrylic resin latex and the No. II acrylic resin in the enteric layer in the step (2) is 1:4, and the rest is consistent with the example 1.

Comparative example 11

The difference between the comparative example and the example 1 is that the mass ratio of the No. I acrylic resin latex and the No. II acrylic resin in the enteric layer in the step (2) is 1:3, and the rest is consistent with the example 1.

Comparative example 12

The difference between the comparative example and the example 1 is that the mass ratio of the No. I acrylic resin latex and the No. II acrylic resin in the enteric layer in the step (2) is 1:10, and the rest is consistent with the example 1.

Comparative example 13

The difference between this comparative example and example 1 is that the acrylic resin No. I in the enteric layer in step (2) is replaced with acrylic resin No. III, and the rest is the same as example 1.

Comparative example 14

The difference between this comparative example and example 1 is that the ratio of the mass of diethyl phthalate to the sum of the mass of acrylic resin No. i and acrylic resin No. ii in the enteric layer in step (2) was 0.1:1, and the rest was kept the same as example 1.

Comparative example 15

The difference between this comparative example and example 1 is that the ratio of the mass of diethyl phthalate to the sum of the mass of acrylic resin No. i and acrylic resin No. ii in the enteric layer in step (2) was 0.3:1, and the rest was kept the same as example 1.

Comparative example 16

The difference between this comparative example and example 1 is that the mass of the enteric layer of step (2) is 6% of the mass of the tablet core, and the rest is in agreement with example 1.

Test example 1

Respectively loading 175ml of the macroporous adsorption resin treated in the example 1 and the comparative examples 1-4 into a column by a wet method, respectively adding an equal amount of gardenia liquid medicine, loading the liquid medicine at a flow rate of 7ml/min, washing 350ml of the liquid medicine with water to remove impurities, eluting 700ml of 60% ethanol at a flow rate of 7ml/min to obtain 60% ethanol elution parts, and determining by UPLC. Then, 350ml of 95% ethanol was added for elution to obtain 95% ethanol-eluted fractions, which were subjected to UPLC measurement. The content determination method comprises the following steps:

a chromatographic column: welch C18 (2.1X 100mm, 1.8 μm);

flow rate: 0.4mL/min, column temperature: 30 ℃; sample introduction volume: 1 μ L, detection wavelength: 238 nm;

acetonitrile (A) -water (B) is used as a mobile phase, 0min, 6% of A; 2.3-4.2min, 6% -12% A; 4.2-5.8min, 12% -17% A; 5.8-8.1min, 17% -20% A; 20-27% A for 8.1-10.4 min; 10.4-15 min, 27% -32% A; 15-16.2 min, 32-36% A; 16.2-16.7 min, 36% -55% A; 16.7-17.8 min, 55% -70% A; 17.8-18 min, 70% -6% A; 18-20 min, 6% A. The results of the content measurement are shown in Table 2.

The results are shown in Table 1.

TABLE 1 macroporous adsorbent resin test results

Test example 2

The tablet cores of examples 1-3 and comparative examples 6-9 were subjected to disintegration time test, and each group of tablet cores was placed in a closed container having an air humidity of 95% +/-2% at 25 ℃ with reference to the Chinese pharmacopoeia, and the results of measuring the moisture absorption rate on day 5 are shown in Table 2.

TABLE 2 disintegration time test results

Test examples	hardness/N	Disintegration time	5 days the core moisture absorption rate%
				Example 1	95	5.0min	3.2
Example 2	95	4.5min	3.8
				Example 3	95	4.8min	4.0
Comparative example 6	107	9.8min	5.8
				Comparative example 7	102	8.0min	3.0
Comparative example 8	98	9.3min	6.1
				Comparative example 9	97	6.5min	7.2

Test example 3

The gardenia duodenal targeting formulations prepared in examples 1 to 3 and comparative examples 10 to 16 were subjected to a release degree test in artificial gastric juice and artificial small intestinal juice.

The media were formulated as follows:

artificial gastric juice (pepsin-free): adding 16.4ml of dilute hydrochloric acid, adding water to 1000ml, shaking up, and degassing by an ultrasonic method after the preparation is finished to obtain the product.

Artificial small intestine juice (without pancreatin): taking 250mL of 0.2mol/L potassium dihydrogen phosphate solution, adding 118mL of 0.2mol/L sodium hydroxide solution, diluting with water to 1000mL, shaking up, and degassing by an ultrasonic method after preparation is finished to obtain the potassium dihydrogen phosphate.

The experimental method comprises the steps of adjusting an instrument according to experimental requirements, after the temperature of a dissolution medium is constant at (37 +/-1) DEG C, respectively placing 6 test samples in each experimental example group into a glass tube of a hanging basket, checking in artificial gastric juice for 2 hours, continuously taking out the hanging basket, washing with a small amount of water, adding 1 baffle plate into each tube, and checking in the artificial small intestine juice according to the method. The disintegration time and appearance results are shown in Table 3.

TABLE 3 Effect of disintegration time (%)

Test example 4 pharmacodynamic test

Experimental methods

(1) Laboratory animals and groups

SD rats, SPF grade, male. After arrival, the test was first carried out 3 days after acclimatization observation in the animal house (barrier system). Animals were randomized into 9 groups of 10 animals each: 10ml/kg (dose of 6.35ml/kg) of Yinzhihuang oral liquid for a control group, a model group and a positive group, a low dose group (dose of 42.5mg/kg) in example 1, a medium dose group (dose of 85.0mg/kg) in example 1, a higher dose group (dose of 170.0mg/kg) in example 1, a high dose group (dose of 340.0mg/kg) in example 1, a comparative example 1 (dose of 42.5mg/kg) and a comparative example 12 (dose of 42.5 mg/kg).

(2) Content of the experiment

Except for the model group and the control group, the other groups are subjected to intragastric administration for 1 time per day according to the test substances and the dosage in the groups, and the administration volume is 10 ml/kg; the model group and the control group are administered with 10ml/kg purified water by intragastric administration. Each group was administered by continuous gavage for 5 days. The control group and the model group were given equal volume of purified water. After 1h on day 3, each group was administered with a single gavage of 8mg/ml α -naphthalene isothiocyanate (ANIT) peanut oil solution 40mg/kg cholestatic liver disease (16 h before ANIT gavage and 4h after gavage) except for the control group. The control group was given the same volume of peanut oil. After 1h of the last administration, 1.0% sodium pentobarbital 50mg/kg was anesthetized by intraperitoneal injection, blood was collected from the abdominal aorta, allowed to stand at room temperature, centrifuged at 3500rmp for 15min, the serum was separated, and alkaline phosphatase (ALP), gamma-transglutaminase (GGT), Total Bile Acid (TBA), and Total Bilirubin (TBIL) in the serum were detected. The results are shown in tables 4 and 5.

TABLE 4 influence of Gardenia duodenum localization preparation on ALP and GGT in ANIT-induced cholestatic liver disease rats: (

n＝10)

Note: the different letters in the same column represent the corresponding groups with significant statistical significance

TABLE 5 Effect of Gardenia duodenum localization preparation on TBA and TBIL in ANIT-induced cholestatic liver injury rats: (

n＝10)

Note: the different letters in the same column indicate that the corresponding groups have significant statistical significance.

Percent TBA reduction ═ model group TBA-experimental group TBA)/model group TBA × 100

Percent reduction of TBIL ═ model group TBIL-experimental group TBIL)/model group TBIL × 100

And (4) analyzing results: serum ALP, GGT, TBA and TBIL of the model group rats are obviously increased, which indicates that the model group rats have obvious cholestatic liver injury. The gardenia duodenum positioning preparation can reduce the level of serum Total Bilirubin (TBIL) and Total Bile Acid (TBA) to different degrees when 4 dosage groups are continuously administrated for 5 days. The percentage of TBIL reduction of 42.5, 85.0, 170.0 and 340.0mg/kg dose groups of gardenia preparation is 63.29 percent, 57.85 percent, 64.19 percent and 89.89 percent respectively; the percentage reduction in TBA was 25.49%, 38.63%, 45.34%, 87.38%, respectively. The results suggest that duodenal localization preparation can significantly reduce cholestasis, thereby reducing TBIL and TBA in serum. In addition, 4 dosage groups of the duodenum positioning preparation can reduce ALP and GGT levels in serum to different degrees, which shows that the gardenia preparation can obviously reduce liver injury. The experimental result suggests that the duodenum positioning preparation has a significant improvement effect on cholestatic liver diseases. Comparative example 1 was purified using different resins and comparative example 12 group used different enteric coating formulations, and the final formulation was less effective than the low dose group of example 1.

The above description is only for the preferred embodiments of the present invention, but these embodiments are only exemplary and do not limit the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.

Claims

1. A preparation method of gardenia extract for treating cholestatic liver diseases is characterized by comprising the following steps:

(1) adding water into fructus Gardeniae to obtain water decoction;

(2) passing the water decoction through SP825L macroporous resin column at the speed of 2-4BV/h, washing with water, eluting with ethanol, collecting ethanol eluate, concentrating, and drying.

2. The preparation method according to claim 1, wherein the mass ratio of the volume of the added water to the gardenia in the step (1) is 10-15:1 mL/g; the decoction is carried out for 2 times, and the time for each decoction is 0.5-2 h.

3. The method according to claim 1, wherein the water washing in the step (2) is water washing with 1-3BV, and the ethanol elution is 30-70% ethanol elution with 3-5 BV.

4. The preparation method of claim 1, wherein the diameter-height ratio of the SP825L macroporous resin column in the step (2) is 1:1.3-1:9.2, and the mass ratio of the crude drug in the water decoction to the SP825L macroporous resin column is 1: 1-3.

5. The preparation method according to claim 1, wherein the concentration in the step (2) is to concentrate the alcohol eluent to an extract with a relative density of 1.21-1.25 at 60 ℃; the drying is spray drying or vacuum drying, the inlet temperature of the spray drying is 85-190 ℃, and the outlet temperature is fixed at 75-100 ℃.

6. The method as claimed in claim 1, wherein the content of geniposide, genipin gentiobioside and hydroxygardenianoside in the obtained gardenia extract is respectively 400-750mg/g, 90-150mg/g and 10-50 mg/g.

7. The gardenia duodenum positioning preparation is characterized by comprising a tablet core and an enteric coating wrapping the tablet core, wherein the tablet core comprises the following components in parts by weight: 35-45 parts of gardenia extract according to any one of claims 1 to 6, 50-70 parts of lactose, 15-25 parts of microcrystalline cellulose, 0.15-0.35 part of magnesium stearate; the enteric coating comprises No. I acrylic resin, No. II acrylic resin and diethyl phthalate.

8. The gardenia duodenal localization preparation as claimed in claim 7, wherein the mass ratio of lactose to microcrystalline cellulose is 2-4:1, and the mass ratio of the acrylic resin I to the acrylic resin II is 1: 4.5-5.5.

9. The gardenia duodenal positioning preparation as claimed in claim 7, wherein the mass of the enteric coating is 8-12% of that of the tablet core; the ratio of the mass of diethyl phthalate to the total mass of the No. I acrylic resin and the No. II acrylic resin is 0.15-0.25: 1.

10. use of the gardenia extract obtained by the preparation method according to any one of claims 1 to 6 or the gardenia duodenal localization preparation according to any one of claims 7 to 9 in preparation of a medicament for treating cholestatic liver disease.