Application of ursodesoxycholic acid in preparation of medicine for treating biliary tract diseases of pets

Abstract

The invention belongs to the technical field of application of veterinary chemical drugs, and provides application of ursodesoxycholic acid in preparation of drugs for treating biliary tract diseases of pets. The medicine for treating the biliary tract diseases of the pets is a medicinal preparation consisting of an active ingredient ursodesoxycholic acid and one or more medicinal carriers. The invention also provides an ursodesoxycholic acid self-microemulsion preparation. The ursodesoxycholic acid self-microemulsion preparation comprises the following components: ursodesoxycholic acid, a surfactant, a cosurfactant, an oil phase and a flavoring agent. After being orally taken, the self-microemulsion preparation is in contact with gastrointestinal fluid and spontaneously emulsified into O / W microemulsion with the particle size smaller than 50 nm, bitter fat-soluble medicine is wrapped in emulsion droplets, contact with taste buds is reduced, and palatability is improved. The self-microemulsion preparation can also cause secretion of bile and lipase of the pets, so that distribution of ursodesoxycholic acid in the bile is promoted, and the curative effect of ursodesoxycholic acid on the aspect of treating biliary tract diseases of the pets is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of veterinary drug application, and mainly relates to the use of ursodeoxycholic acid in preparing a medicine for treating pet biliary diseases. Background Art

[0002] Ursodeoxycholic acid is an odorless, bitter white powder, a stone dissolving agent, used to prevent and treat gallstones. It is also used to treat various liver pathological conditions such as hepatic cholestasis and primary biliary cirrhosis. Ursodeoxycholic acid is almost insoluble in water and is a poorly soluble drug, which limits its clinical application. The drug is easily soluble in ethanol, insoluble in chloroform; easily soluble in glacial acetic acid, and soluble in sodium hydroxide test solution. Ursodeoxycholic acid is a multifunctional hydrophilic dihydroxycholic acid, with the chemical name 3α,7β-dihydroxy-5β-cholestane-24-acid, which accounts for about 3% of the total bile acid in the normal human body. Ursodeoxycholic acid is the only non-surgical bile acid therapy approved by the US FDA for the treatment of bile acid, with the advantages of high efficacy and no side effects.

[0003] Ursodeoxycholic acid has good pharmacological and clinical effects. The pharmacological effects are mainly reflected in protecting cells, changing bile acid composition, regulating the immune system, and reducing lipids. In addition, because ursodeoxycholic acid can significantly reduce serum cholesterol and triglycerides. Ursodeoxycholic acid is an ideal drug for the treatment of hyperlipidemia. In recent years, research reports on the treatment of fatty liver with ursodeoxycholic acid have laid the foundation for its application and provided clinical basis. Studies have shown that ursodeoxycholic acid is similar to the cholesterol resin cholestyramine, and can significantly reduce serum cholesterol, triglycerides and LDL-C levels, increase serum HDL-C levels, and reduce lipid deposition in the liver, improve liver vitality, stabilize liver cell membranes and inhibit monocytes from producing cytokines. In addition, ursodeoxycholic acid can also play a certain protective role, promote cell metabolism, promote the transportation and removal of fat, and on the other hand, it can also reduce blood lipids, increase blood lipid excretion, and reduce lipid intake.

[0004] Ursodeoxycholic acid can alleviate the patient's clinical symptoms, and can effectively improve the CT signs of the liver, and has a good effect on fatty liver. At the same time, in terms of clinical effects, ursodeoxycholic acid shows better application prospects. There are many trials that have shown that ursodeoxycholic acid is beneficial for various liver pathologies, however, its effect on cholelithiasis is most significant. Currently, ursodeoxycholic acid has been confirmed to be an effective treatment for cholecystectomy in patients with gallstones. Ursodeoxycholic acid is usually used to treat patients with primary biliary cirrhosis, which is a biliary liver disease induced by the toxic effects of bile acid stasis and secondary bile duct destruction caused by immune abnormalities. However, the therapeutic effect of ursodeoxycholic acid on pet hepatobiliary diseases is still unclear.

[0005] In the prior art, ursodeoxycholic acid drugs are capsules and tablets. After oral administration, they are mainly absorbed in the upper jejunum by non-ionic passive diffusion and in the ileum by active transport. After absorption, they are mainly distributed in the hepatic and intestinal tissues and plasma, and the plasma protein binding rate is 96-99%. Its therapeutic effect mainly depends on the concentration of ursodeoxycholic acid in bile. Ursodeoxycholic acid tastes very bitter. In human medicine, it is generally made into capsules, or coated tablets, or a large amount of flavoring agents or sweeteners are added to meet the needs of patients. However, the dosage forms and specifications of existing human medicines are not suitable for clinical application in pets.

[0006] In the clinical application of pets, due to the large number of individual differences among pets, it is often necessary to develop products of different specifications to meet the needs of animals of different weights, which increases the costs of various stages such as research and development, production, quality control, and packaging. For example, if it is developed into an oral solution dosage form, it can be dispensed with a dispenser, and animals can be precisely dosed according to their weight, which has better clinical safety and effectiveness. However, ursodeoxycholic acid is a poorly soluble drug, which limits the possibility of developing an oral solution.

[0007] Therefore, it is necessary to study the therapeutic effect of ursodeoxycholic acid on pet hepatobiliary diseases and to develop an oral preparation of ursodeoxycholic acid that is suitable for clinical application in pets and convenient for administration. Summary of the invention

[0008] In order to solve the problems in the prior art and overcome the technical defects, the present invention provides the use of ursodeoxycholic acid in a drug for treating pet biliary diseases.

[0009] In a first aspect, the present invention provides use of ursodeoxycholic acid in preparing a medicament for treating biliary tract diseases in pets.

[0010] The medicine for treating pet biliary diseases is a pharmaceutical preparation composed of an effective ingredient, ursodeoxycholic acid, and one or more pharmaceutical carriers.

[0011] Preferably, the pharmaceutical preparation is an ursodeoxycholic acid self-microemulsification preparation.

[0012] Preferably, the pet biliary diseases include cholestasis, bile duct stones, bile duct obstruction, cholecystitis, and cholangitis.

[0013] More preferably, the pet biliary disease is cholecystitis.

[0014] In a second aspect, the present invention provides an ursodeoxycholic acid self-microemulsifying preparation, wherein the self-microemulsifying preparation comprises the following components: ursodeoxycholic acid, a surfactant, a co-surfactant, an oil phase, and a flavoring agent.

[0015] The dosage of each component in every 100 ml of the self-microemulsification preparation is: 5.0 g of ursodeoxycholic acid, 30-50 ml of surfactant, 30-50 ml of oil phase, 0-3 mg of flavoring agent, and the remainder is co-surfactant.

[0016] Preferably, the dosage of each component in every 100 ml of the self-microemulsification preparation is: 5.0 g of ursodeoxycholic acid, 35-45 ml of surfactant, 35-45 ml of oil phase, 1-2 mg of flavoring agent, and the remainder is co-surfactant.

[0017] Preferably, the ratio of the oil phase to the surfactant is (8-9):(7-8).

[0018] More preferably, the ratio of the oil phase to the surfactant is 1:1.

[0019] The surfactant is one or more of Tween 80, polyoxyethylene hydrogenated castor oil, caprylic capric macrogol glyceride, caprylic capric mono- and di-glyceride, polyoxyethylene glyceride, and propylene glycol monocaprylate.

[0020] Preferably, the surfactant is one or more of polyoxyethylene hydrogenated castor oil, caprylic capric macrogol glyceride, caprylic capric mono- and di-glyceride.

[0021] More preferably, the surfactant is polyoxyethylene 40 hydrogenated castor oil.

[0022] The co-surfactant is one or more of polyethylene glycol, propylene glycol, ethanol, and diethylene glycol monoethyl ether.

[0023] Preferably, the co-surfactant is selected from ethanol and diethylene glycol monoethyl ether.

[0024] More preferably, the co-surfactant is ethanol.

[0025] The oil phase is one or more of corn oil, soybean oil, cottonseed oil, modified corn oil, monoolein, monolinolein, medium chain triglyceride, and oleic acid.

[0026] Preferably, the oil phase is one or more of monoolein, monolinolein, medium chain triglycerides, and oleic acid.

[0027] More preferably, the oil phase is medium chain triglycerides.

[0028] The flavor corrector is one or more of oil-soluble meat flavor, oil-soluble milk flavor, oil-soluble seafood flavor, animal hydrolyzed protein, brewer's yeast, sucrose, and sucralose.

[0029] Preferably, the flavoring agent is an oil-soluble meat flavoring.

[0030] A more preferred formulation is: per 100 ml of the self-microemulsification preparation, the dosage of each component is 5.0 g of ursodeoxycholic acid, 40 ml of polyoxyethylene 40 hydrogenated castor oil, 40 ml of medium chain triglyceride, 1 mg of flavoring agent, and the balance is ethanol.

[0031] The third aspect of the present invention provides a method for preparing an ursodeoxycholic acid self-microemulsification preparation, the steps of which are as follows:

[0032] (1) Add ursodeoxycholic acid and flavoring agent to the oil phase and stir to dissolve evenly;

[0033] (2) adding a surfactant and a co-surfactant, stirring slightly at room temperature until dissolved, and filtering to obtain a liquid;

[0034] (3) Fill the liquid obtained in step (2) into a medicinal glass bottle, add a stopper, and screw the cap.

[0035] Preferably, in step (4), after the liquid is filled into a pharmaceutical glass bottle, the headspace is filled with nitrogen for protection.

[0036] In a fourth aspect, the present invention provides a use of an ursodeoxycholic acid self-microemulsification preparation in the preparation of a medicament for treating pet biliary diseases.

[0037] The pets include but are not limited to dogs, cats, rabbits, and horses.

[0038] Preferably, the pet is selected from dogs and cats.

[0039] Beneficial effects:

[0040] 1. The self-microemulsification prescription of the present invention is a lipid prescription. The drug contains unsaturated long-chain fatty acids. After contacting with body fluids, chylomicrons are formed to combine the drug with lipoproteins, promote lymphatic absorption, and reduce the first-pass effect of the liver. Secondly, the intake of lipid prescriptions can cause the secretion of bile and lipase in pets, thereby promoting the distribution of ursodeoxycholic acid in bile and improving the efficacy of the drug.

[0041] 2. The self-microemulsification formulation of the present invention can be spontaneously emulsified into an O / W microemulsion with a particle size of less than 50 nm after contacting with saliva and gastrointestinal fluid in the oral cavity and after being taken. The bitter fat-soluble drug is encapsulated in the emulsion droplets, which reduces the contact with taste buds and improves the compliance of pets. The ursodeoxycholic acid self-microemulsion prepared by the present invention is easier for pet dogs and cats to take when treating hepatobiliary diseases than tablets and capsules, and has better palatability.

[0042] 3. The present invention adopts a self-microemulsification formulation to dissolve ursodeoxycholic acid in the oil phase or as a surfactant. Under the action of the surfactant, ursodeoxycholic acid spontaneously forms emulsion particles after encountering gastrointestinal fluid, thereby increasing the absorption of the drug; at the same time, ursodeoxycholic acid is encapsulated in the emulsion, which has better stability.

[0043] 4. The ursodeoxycholic acid self-microemulsion prepared by the present invention has a relatively simple preparation process, and the raw and auxiliary materials are relatively easy to obtain, and the requirements for production equipment are not high, and the feasibility of large-scale production is high. DETAILED DESCRIPTION

[0044] Example 1

[0045] A ursodeoxycholic acid self-microemulsification preparation is prepared by the following method:

[0046] Element Single dose (100ml / bottle) Ursodeoxycholic acid 5.0g Medium Chain Triglycerides 45ml Polyoxyethylene 40 Hydrogenated Castor Oil 35ml Oil-soluble meat flavor 1mg Ethanol Refill to 100ml

[0047] Under stirring conditions, ursodeoxycholic acid and oil-soluble meat flavor are first dissolved in medium-chain triglycerides, and then the prescribed amount of polyoxyethylene 40 hydrogenated castor oil is added and mixed evenly. Finally, ethanol is used to make the volume constant, mixed evenly, filtered, and packaged.

[0048] Example 2

[0049] A ursodeoxycholic acid self-microemulsification preparation is prepared by the following method:

[0050]

[0051]

[0052] Under stirring conditions, ursodeoxycholic acid and oil-soluble meat flavor were first dissolved in medium-chain triglycerides, and then polyoxyethylene 40 hydrogenated castor oil was added and mixed evenly. Finally, ethanol was used to make the volume constant, mixed evenly, filtered, and packaged. One part of the samples was directly packaged as a non-nitrogen-filled group, and another part of the samples was packaged and the headspace was filled with nitrogen for protection as a nitrogen-filled group.

[0053] Example 3

[0054] A ursodeoxycholic acid self-microemulsification preparation is prepared by the following method:

[0055] Element Single dose (100ml / bottle) Ursodeoxycholic acid 5.0g Medium Chain Triglycerides 40ml Polyoxyethylene 40 Hydrogenated Castor Oil 40ml Oil-soluble meat flavor 1mg Ethanol:polyethylene glycol (1:1) Refill to 100ml

[0056] Under stirring conditions, ursodeoxycholic acid and oil-soluble meat flavor are first dissolved in medium-chain triglycerides, and then polyoxyethylene 40 hydrogenated castor oil is added and mixed evenly. Finally, an ethanol: polyethylene glycol (1:1) mixed solution is used to make the volume, mixed evenly, filtered, and packaged.

[0057] Example 4

[0058] An ursodeoxycholic acid self-microemulsion is prepared by the following method:

[0059] Element Single dose (100ml / bottle) Ursodeoxycholic acid 5.0g Medium Chain Triglycerides 35ml Polyoxyethylene 40 Hydrogenated Castor Oil 45ml Oil-soluble meat flavor 1mg Ethanol Refill to 100ml

[0060] Under stirring conditions, ursodeoxycholic acid and oil-soluble meat flavor are first dissolved in medium-chain triglycerides, and then polyoxyethylene 40 hydrogenated castor oil is added and mixed evenly. Finally, an ethanol: polyethylene glycol (1:1) mixed solution is used to make the volume, mixed evenly, filtered, and packaged.

[0061] Comparative Example 1

[0062] Ursodeoxycholic acid capsules are now available for human use. The specification is 250 mg / capsule and the trade name is Ursof.

[0063] Comparative Example 2

[0064] An ursodeoxycholic acid self-microemulsion is prepared by the following method:

[0065] Element Single dose (100ml / bottle) Ursodeoxycholic acid 5.0g Monolinoleylglycerol 40ml Caprylic / capric macrogol glycerides 40ml Oil-soluble meat flavor 1mg Ethanol Refill to 100ml

[0066] Under stirring conditions, ursodeoxycholic acid and oil-soluble meat flavor are first dissolved in monolinoleyl glyceride, and then caprylic acid capric acid macrogol glyceride is added and mixed evenly. Finally, ethanol is used to make up the volume, mixed evenly, filtered, and packaged.

[0067] Comparative Example 3

[0068] An ursodeoxycholic acid self-microemulsion is prepared by the following method:

[0069] Element Single dose (100ml / bottle) Ursodeoxycholic acid 5.0g Monolinoleylglycerol 50ml Polyoxyethylene 40 Hydrogenated Castor Oil Refill to 100ml

[0070] Under stirring conditions, first dissolve ursodeoxycholic acid in monolinoleylglycerol, then add polyoxyethylene 40 hydrogenated castor oil to make up the volume, mix evenly, filter and package.

[0071] Example 5 Investigation of emulsification rate and droplet size

[0072] The emulsification phenomenon and the droplet particle size after emulsification were investigated for Example 1, Example 2, Example 3, Example 4 and Comparative Example 2 and Comparative Example 3.

[0073] Table 1 Investigation of emulsification rate and droplet size

[0074]

[0075] The test shows that the droplet size in Example 1 is relatively large, and it is not clear after dilution with water, and is a slightly turbid liquid. The reason for this may be that the amount of surfactant used is small. In Comparative Example 2, caprylic acid capric acid macrogol glyceride is used, and the emulsification effect is poor, and the prepared droplet size is relatively large. In Comparative Example 3, because no co-surfactant is added, the droplet stability is poor, and stratification occurs after being placed for 24 hours.

[0076] Example 6 Stability Study

[0077] Under accelerated conditions (40°C ± 2°C, RH 75% ± 5%), samples of Examples 2, 3, 4 and Comparative Example 2 were respectively packaged in 100 ml glass bottles, sealed, and placed for 6 months. The properties and content changes are shown in Table 2:

[0078] Table 2 Acceleration conditions results

[0079]

[0080] The test results show that during the acceleration process, the properties of each sample changed, among which the color of Comparative Example 2 changed significantly. The reason may be that the monolinoleic acid glyceride used in the oil phase is unstable and easily oxidized; the properties of Example 2 (non-nitrogen-filled group), Example 3, and Example 4 groups changed slightly, which shows that the medium-chain triglyceride in the oil phase used in Examples 2, 3, and 4 has better oxygen stability. The properties of Example 2 (nitrogen-filled group) did not change significantly because the nitrogen filling of its headspace isolated the oxidation range of oxygen on the oil phase and had better stability.

[0081] Example 7 Pharmacokinetic Test

[0082] Example 2 and Comparative Example 1 were selected to compare the pharmacokinetics in beagle dogs.

[0083] Experimental animals: Twelve healthy male beagle dogs were selected and randomly divided into two groups, with 6 dogs in each group.

[0084] Dosage: Oral administration at 10 mg / kg body weight, fasting.

[0085] Sampling points: Blood was collected for testing before administration and 15min, 30min, 45min, 1h, 2h, 4h, 6h, 8h, 12h, 24h, 36h, 48h, 72h, 96h, and 120h after administration, and bile was collected at 2h, 6h, 12h, 24h, and 48h to determine the drug content in bile. The main pharmacokinetic parameters are shown in Table 3.

[0086] Table 3 Pharmacokinetic parameters and bile concentration data of Example 2 group and Comparative Example 1 group after administration

[0087] Project Name unit Example 2 Group Comparative Example 1 <![CDATA[C max ]]> μg / ml 5.76±6.35 5.42±2.62 AUC (μg / ml)·h 1113±279 726±478 Drug content in bile μmol / L 2.53 1.41

[0088] The results show that the AUC of Example 2 group is higher than that of Comparative Example 1 group, indicating that ursodeoxycholic acid has higher bioavailability after being prepared into self-microemulsion compared with ordinary capsules. It is particularly noteworthy that the drug concentration in bile of Example 2 is 179% of that of Comparative Example 1, which may be due to the fact that the self-microemulsion prescription can increase the bioavailability of the drug, or the lipid excipients in the formula can promote the secretion of bile.

[0089] Example 8 Palatability Test

[0090] The capsules of Example 2, Comparative Example 3 and Comparative Example 1 of the present invention were compared in terms of administration palatability.

[0091] Administration method: In Example 2 and Comparative Example 3, a certain dose was drawn with a syringe and directly injected into the pet's mouth; in Comparative Example 1, the capsule shell was removed, suspended in water, and then administered, drawn with a syringe and directly injected into the pet's mouth. Judgment criteria: If the pet shows resistance, salivation, head shaking, etc., it is recorded as resistance. If the above phenomena do not occur, the dose is accepted.

[0092] In our animal house, stray base and pet hospital, we collected palatability data of 50 dogs and 50 cats. The results are as follows:

[0093] Table 4 Palatability test of ursodeoxycholic acid self-microemulsion

[0094]

[0095]

[0096] This experiment shows that Example 2 of the present invention has better palatability than Comparative Example 1, and Comparative Example 3 without flavoring agent also has good palatability, but is slightly worse than Example 2 containing flavoring agent.

[0097] Example 9 Clinical Effect Trial on Treatment of Cholecystitis in Dogs

[0098] 20 dogs with gallstones were selected and randomly divided into two groups, one group took ursodeoxycholic acid self-microemulsion, and the other group took ursodeoxycholic acid capsules; the administration method is shown in Table 5. The cure rate and symptoms before and after administration were statistically analyzed. Records were made on the day before administration, the first day of administration, the third day of administration, and the seventh day of administration. From the cure rate, it can be seen that 100% cure can be achieved after 7 days of administration. The improvement and recovery of the ursodeoxycholic acid capsule treatment group were slightly slower than those of the ursodeoxycholic acid self-microemulsion treatment group. The results are shown in Table 6.

[0099] Table 5 Symptoms and cure rates of dogs before and after medication

[0100] Serial number Ursodeoxycholic acid self-microemulsion treatment group (n=10) Ursodeoxycholic acid capsule treatment group (n=10) Dosing frequency Dosage 1 Ursodeoxycholic acid self-microemulsion oral solution 15mg / kg Ursodeoxycholic acid capsules 15mg / kg 1 time / day oral 2 Amoxicillin clavulanate potassium 25mg / kg Amoxicillin clavulanate potassium 25mg / kg 1 time / day oral 3 Urinary Tract Inhibitor 500mg Urinary Tract Inhibitor 500mg 1 time / day oral 4 Danos 20mg / kg Danos 20mg / kg 1 time / day oral 5 Omeprazole injection 1.5mg / kg Omeprazole injection 1.5mg / kg 1 time / day Subcutaneous injection 6 Kotezhuang injection 1ml Kotezhuang injection 1ml 1 time / day Subcutaneous injection

[0101] Table 6 Symptoms and cure rates of dogs before and after medication

[0102]

Claims

1. Use of ursodeoxycholic acid in the preparation of a medicament for treating pet biliary tract diseases, characterized in that: The medicine for treating pet biliary diseases is a pharmaceutical preparation composed of an effective ingredient, ursodeoxycholic acid, and one or more pharmaceutical carriers.

2. The use according to claim 1, characterized in that The pharmaceutical preparation is an ursodeoxycholic acid self-microemulsification preparation.

3. The use according to claim 1, characterized in that The pet biliary diseases include cholestasis, bile duct stones, bile duct obstruction, cholecystitis, and cholangitis.

4. An ursodeoxycholic acid self-microemulsification preparation, characterized in that: The self-microemulsification preparation comprises the following components: ursodeoxycholic acid, a surfactant, a co-surfactant, an oil phase and a flavoring agent.

5. The self-microemulsification preparation according to claim 4, characterized in that The dosage of each component in every 100 ml of the self-microemulsification preparation is: 5.0 g of ursodeoxycholic acid, 30-50 ml of surfactant, 30-50 ml of oil phase, 0-3 mg of flavoring agent, and the remainder is co-surfactant.

6. The self-microemulsification preparation according to claim 5, characterized in that The dosage of each component in every 100 ml of the self-microemulsification preparation is: 5.0 g of ursodeoxycholic acid, 35-45 ml of surfactant, 35-45 ml of oil phase, 1-2 mg of flavoring agent, and the remainder is co-surfactant.

7. The self-microemulsification preparation according to any one of claims 4 to 6, characterized in that The surfactant is one or more of Tween 80, polyoxyethylene hydrogenated castor oil, caprylic capric macrogol glyceride, caprylic capric mono- and di-glyceride, polyoxyethylene glyceride, and propylene glycol monocaprylate.

8. The self-microemulsification preparation according to any one of claims 4 to 6, characterized in that The co-surfactant is one or more of polyethylene glycol, propylene glycol, ethanol, and diethylene glycol monoethyl ether.

9. The self-microemulsification preparation according to any one of claims 4 to 6, characterized in that The oil phase is one or more of corn oil, soybean oil, cottonseed oil, modified corn oil, monoolein, monolinolein, medium-chain triglycerides, and oleic acid; the flavoring agent is one or more of oil-soluble meat flavor, oil-soluble milk flavor, oil-soluble seafood flavor, animal hydrolyzed protein, brewer's yeast, sucrose, and sucralose.

10. Use of the ursodeoxycholic acid self-microemulsification preparation according to any one of claims 4 to 6 in the preparation of a medicament for treating biliary tract diseases in pets.

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