CN1221283C - Oral insulin granule and its preparation - Google Patents

Abstract

The present invention relates to an oral fine insulin granule and a preparation method thereof. The oral fine insulin granule taken orally performs the effect of lowering blood sugar. The oral fine insulin granule is an enteric capsule prepared from insulin gelatin composition and liposome precursor granule, and the encapsulation efficiency for insulin is more than 90%. The preparation method comprises: firstly, insulin and gelatin are prepared into a composition according to a certain proportion; then, the composition is prepared into a plastid particle dispersion system, and a proper quantity of support agent is added in the plastid particle dispersion system for freeze drying; finally, the plastid particle dispersion system with the support agent is pulverized, mixed with a proper quantity of gum arabic, ground and sieved. The present invention is suitable for diabetic patients and avoids side effects, such as lipoatrophy, etc., caused by long-term injection.

Description

Oral insulin microparticles and preparation method thereof

The invention relates to an oral insulin microgranule and a preparation method thereof. It is a new dosage form of the hypoglycemic drug insulin. To explore a new way for oral administration of biochemical drugs.

Insulin is a protein polypeptide drug, which can be degraded by proteolytic enzymes in the gastrointestinal tract, and is not absorbed. It is ineffective when taken orally, and it is generally administered by injection. Diabetic patients need long-term injections, which is very inconvenient and causes subcutaneous fat atrophy. Non-injection dosage forms such as oral administration are being studied at home and abroad. Oral administration dosage forms mainly focus on the following aspects: 1) Enteric-coated preparations: the British patent absorbs insulin on cellulose and glucose gel particles, and coats them with enteric coating. For oral administration (GB 1987, 2180746), Saffran coats insulin microparticles and packs them into enteric-coated capsules (Science, 1987, 233: 1081). 2) Adding enzyme inhibitors or protective agents: Zir et al. have developed insulin tablets containing soybean trypsin inhibitors (J Pharm Sic, 1994, 83(6): 792); Japanese patents introduce that mucin can protect insulin from intestinal Dissolving capsules (JP 1979, 54028807 (A)). 3) Adding absorption enhancers: Touitou et al. made a mixture preparation of Situ Mage 1000 and insulin (J Pharm Pharmacol1980, 32:108). 4) Insulin microcapsules and nanospheres: Insulin nanocapsules made by Fallouh et al. (IntJ Pharm, 1986, 28(2): 125), domestic Zhang Qiang et al. Certain hypoglycemic effect (Acta Pharmacologica Sinica, 1998, 32(2): 152). 5) Insulin liposome: Canadian patent uses liposome to encapsulate insulin to make oral preparation (CA, 1994, 1328401). 6) Insulin emulsion: Smithkline Beecham Company systematically developed insulin emulsion (WO, 1994, 9408603; US, 1993, 5179079; WO, 1994, 9408604); domestic Wu Qiongzhu and others made insulin into multilayer emulsion (W/O/ W) intragastric administration to mice at a dose of 70 U/kg has hypoglycemic effect (Chinese Journal of Pharmaceutical Industry, 1990, 21(10): 445). To sum up, although there are many research reports and patents, they have not been clinically applied so far. The main problem is that the oral administration dose is too high and the bioavailability is low, so it is difficult to apply it clinically.

The object of the present invention is to combine insulin gelatin complex and liposome preparation technology to prepare oral insulin microparticles and pack them into enteric-coated capsules for oral use.

The purpose of the present invention is achieved through the following scheme: the oral insulin microparticles are composed of insulin gelatin complex and liposome precursor microparticles, and the encapsulation rate of insulin is above 90%. First, insulin is made into a complex with gelatin. The consumption weight ratio of insulin and gelatin is 1: 1～5, and the solution concentration insulin is 5～10%, and gelatin is 1～6%, and pH value should be controlled at 5～6, and binding rate is 85%; Liposome microparticle dispersion system, preparation of phospholipid mixed surfactant: soybean lecithin 1.2-2.0, cholesterol 0-0.5, poloxamer 0.2-1.0, chlorobutanol 0.1-0.2, add ethanol to dissolve, reduce pressure, recover Ethanol, to make phospholipid mixed surfactant, the dosage ratio of insulin-gelatin complex and mixed surfactant is 1:5-10; then, make insulin-gelatin complex liposome dispersion system, add appropriate amount of support agent microcrystalline cellulose , Micropowder silica gel, etc. are then freeze-dried, preferably micropowder silica gel. The above-mentioned product was frozen at -18°C for 24 hours, then freeze-dried, and the dried particles were agglomerated. After crushing and adding an appropriate amount of micro-powdered silica gel, they were ground through a 32-mesh sieve.

The invention has the advantages of reducing the inactivation of insulin, appropriate oral administration dose, and high bioavailability. When the oral administration dose is 8 U/kg to model mice, the percentage rate of blood sugar reduction is 33%, and the effect on hyperglycemia rabbits is , Dogs with normal blood sugar all have hypoglycemic effect, so it has a wide range of clinical application prospects. Side effects such as subcutaneous fat atrophy caused by long-term injection administration in diabetic patients can be avoided.

The present invention will be further described in detail below in conjunction with the examples.

The preparation process steps of insulin microparticles are as follows:

1. Preparation of insulin-gelatin complex:

1) Add 10 ml of hydrochloric acid solution (pH1-2) to dissolve 1 g of insulin, and adjust the pH value to 4 with sodium hydroxide solution (10%-20%) after dissolving.

2) Add 1g-2g of gelatin to 50-100ml of distilled water to dissolve into a gelatin solution.

3) Add the insulin solution into the gelatin solution under stirring, mix evenly, adjust the pH to 5-8 with 10% NaOH solution, and the complex is precipitated to form an insulin-gelatin complex suspension to protect the stability of insulin.

2. Preparation of insulin-gelatin complex liposomes

1) Phospholipid mixed surfactants are added to the insulin-gelatin complex suspension, the purpose is to wrap the complex in liposomes, and disperse for 3 minutes at 10,000 rpm to form complex liposomes, which can further enhance the stability of insulin Sex and promote absorption.

2) Preparation of phospholipid mixed surfactant: add 2g cholesterol, 2g-5g poloxamer, 0.5-1g chlorobutanol to 12g soybean lecithin, then add absolute ethanol to dissolve into a solution, recover ethanol under reduced pressure, pump The phospholipid mixed surfactant is dry prepared, and the ratio of its dosage to the compound is 5-10:1.

3. Preparation of insulin microparticles

1) Insulin-gelatin complex liposomes are added with appropriate amount of support agent microcrystalline cellulose and micropowder silica gel, stirred and dispersed evenly, frozen in the freezer for 24 hours, and then freeze-dried.

2) Grind and pulverize the dried agglomerates in a mortar: add an appropriate amount of micropowder silica gel to absorb the residual water, pass through a 32-mesh sieve, put into enteric-coated capsules, seal, and freeze in the refrigerator.

Example 1:

1. Dissolve 0.1 g of insulin in 1.0 ml of pH 2 hydrochloric acid solution, and then adjust the pH to 4 with 10% NaOH solution to make an insulin solution.

2. Dissolve 0.1g of gelatin in 5.0ml of water to form a solution. Under stirring, add insulin solution and mix to form a solution. Adjust the pH to 5.8 with 10% NaOH solution. Precipitation occurs, which is an insulin-gelatin complex, forming a complex turbid solution.

3. Add 1.7g of phospholipid mixed surfactant to the insulin-gelatin complex suspension (composition: 1.2g of soybean lecithin, 0.2g of cholesterol, 0.2g of poloxamer, 0.1g of chlorobutanol, dissolve and mix with ethanol, reduce Press, reclaim ethanol to make phospholipid mixed surfactant), disperse for 3 minutes through the mash machine, add micropowder silica gel 1.0g again, stir and disperse the liposome dispersion system evenly.

4. Freeze the prepared insulin-gelatin complex and liposome dispersion in a freezer (-18° C.) for 24 hours, and then freeze-dry.

5. Grind the dried solid in a mortar, add desiccant micro-powdered silica gel to form microparticles, pass through a 32-mesh sieve to obtain insulin microgranules, and measure the potency. Seal and store in the refrigerator.

Example 2:

1. Dissolve 0.2 g of insulin in 1.0 ml of pH 2 hydrochloric acid solution, adjust the pH to 4 with 10% NaOH solution after dissolving, and make insulin solution.

2. Add 0.2g of gelatin to 5.0ml of water and heat to dissolve, let it cool to 20°C, add the prepared insulin solution under stirring, mix evenly to form a solution, adjust the pH to 5.8 with 10% NaOH solution, precipitate and form insulin gelatin compound things.

3. Add 3.4 g of phospholipid mixed surfactant (composition: 2.4 g of soybean lecithin, 0.4 g of cholesterol, 0.4 g of poloxamer, and 0.2 g of chlorobutanol) into the suspension of insulin-gelatin complex, and grind it into After dispersing at 10000rpm for 3 minutes, add 1g of microcrystalline cellulose, stir and disperse to form a uniform liposome dispersion with low encapsulation efficiency.

4. Freeze the uniformly dispersed system in a freezer (-18°C) for 24 hours, then freeze-dry.

5. Grind and pulverize the dried insulin-gelatin compound particles in a mortar, add an appropriate amount of micropowder silica gel to absorb the residual water into particles, pass through a 32-mesh sieve to make insulin particles, measure the potency, mark, seal, Store in the refrigerator.

Example 3:

1. Add 1 g of insulin to 5 ml of pH 2 hydrochloric acid solution to dissolve. After dissolving, adjust the pH to 4.5 with concentrated NaOH solution to make an insulin solution.

2. Dissolve 1g of gelatin in 50ml of water to form a solution, add the insulin solution dropwise under stirring, mix evenly, adjust the pH to 5.8 with concentrated NaOH solution, and precipitate out to form an insulin-gelatin complex, forming a complex turbid solution.

3. Add 17g of phospholipid mixed surfactant to the insulin gelatin turbid solution (composition: 12g of soybean lecithin, 2g of cholesterol, 2g of poloxamer, 1g of chlorobutanol, add ethanol to dissolve and mix, recover ethanol, and make phospholipid mixed surface Active agent) was dispersed by a pounder at 10,000rpm for 3 minutes, then 10g of micropowdered silica gel was added, stirred, dispersed and evenly mixed to form a dispersion system of insulin gelatin particles.

4. Disperse the microparticles in a freezer (-18°C), freeze for 24 hours, and freeze-dry.

5. Grind and pulverize the dry dispersed particles in a mortar, add appropriate amount of micropowder silica gel to absorb residual water to form particles, pass through a 32-mesh sieve to make insulin microparticles, mark the potency, seal, and store in the refrigerator.

Claims (3)

1. Oral insulin microparticles, characterized in that: the oral insulin microparticles are composed of insulin gelatin complex and liposome precursor microparticles, and the encapsulation rate of insulin is between 90% and 100%. It is an enteric-coated capsule. 2. A method for preparing oral insulin microgranules as claimed in claim 1, characterized in that: 1) Make a complex of insulin and gelatin, add 1g of insulin into 10ml of hydrochloric acid solution with a pH value of 1~2, adjust the pH value to 4 with 10%~20% sodium hydroxide solution after dissolving, and add 1g~2g of gelatin Add 50-100ml of distilled water to dissolve into a gelatin solution, add the insulin solution into the gelatin solution under stirring, mix well, adjust the pH value to 5-8 with 10% sodium hydroxide solution, the complex is precipitated, and an insulin-gelatin complex suspension is formed ; 2) The complex is made into a liposome particle dispersion system, and the phospholipid mixed surfactant is prepared: 12 g soybean lecithin is added with 2 g cholesterol, 2 g ~ 5 g poloxamer, 0.5 ~ 1 g chlorobutanol, and then added absolute ethanol Dissolve into a solution, recover ethanol under reduced pressure, drain to prepare phospholipid mixed surfactant, the ratio of its dosage to the complex is 5-10:1; 3) For the insulin-gelatin complex liposome dispersion system, add an appropriate amount of support agent microcrystalline cellulose and micropowder silica gel, and then freeze-dry. 3. The preparation method of oral insulin microgranules according to claim 2, characterized in that: the finished product is frozen at -18°C for 24 hours, then freeze-dried, the dried particles are agglomerated, crushed and added with appropriate amount of micro-powdered silica gel and ground for 32 hours. Mesh sieve.