NO874852L - PHARMACEUTICAL MIXTURES. - Google Patents

Description

Pharmaceutical mixtures.

Description.

The present invention generally relates to mixtures

for oral administration of protein materials in biologically active form and methods and apparatus for producing these.

More specifically, the present invention relates to compositions for the treatment of diabetes by oral administration of insulin.

Many medicines, drugs and treatments are administered parenterally because they are broken down or not sufficiently absorbed in the stomach and gastrointestinal tract and therefore cannot be administered orally. For example, as discussed in detail below, insulin is administered by subcutaneous injection to many patients suffering from diabetes mellitus.

Diabetes mellitus is a chronic disorder that affects carbohydrate fat and protein metabolism. It is characterized by hyperglycaemia and glycosuria resulting from a deficient or insufficient insulin secretory reaction. Two main variants of the disorder exist. The number of patients diagnosed as diabetic is estimated to be 10 million in the United States alone and this number is expected to increase at a rate of 6% per year.

A variant, seen in approximately 10% of all idiopathic diabetics, is referred to as insulin-dependent diabetes mellitus ("IDDM") or juvenile diabetes. This variant often appears for the first time in youth and is characterized by a gradual loss of the insulin secretion function of the beta cells in the pancreas and thus a progressive "dependence" on insulin of foreign origin to maintain carbohydrate metabolism. (These characteristics are shared by the non-idiopathic, or "secondary", diabetics whose disorders originate in disorders of the pancreas. The other variety of idiopathic diabetes mellitus is referred to as non-insulin-dependent diabetes mellitus ("NIDDM") or age-related -diabetes mellitus and accounts for the rest of the idiopathic diabetic population.

All diabetics, regardless of their genetic and environmental background or the age at the onset of the disorder, have a clear lack of insulin or insufficient insulin function. Because the transfer of glucose from the blood to muscle and fat tissue is insulin-dependent, diabetics lack the ability to utilize the glucose adequately. Furthermore, because glycogenolysis is usually inhibited by insulin, the rate of glycogenolysis is increased in the diabetic. Both of these "deviations" from normal metabolic events lead to the accumulation of glucose in the blood (hyperglycaemia) to the point where the kidneys' reabsorption capacity for glucose is exceeded and glycoseria occurs. The main source of energy for the diabetic is thus fatty acids that come from triglycerides stored in adipose tissue.

In the liver, fatty acids are oxidized to ketone bodies which are circulated and used as an energy source by the tissues. In the IDDM patient, and sometimes the NIDDM patient, the rate of formation of ketone bodies may exceed the rate of their consumption and ketosis together with metabolic acidosis may occur. As the tissues appear to be starved of glucose, diet and tissue sources of protein are used in glucogenesis. Anabolic processes such as synthesis of glycogen, triglycerides and proteins are "sacrificed" for catabolic activities that include glycogenolysis, gluconeogenesis and mobilization of fat. Thus, the diabetic condition originating in a "simple" insulin defect results in extensive metabolic disturbances that have long-term pathological effects on almost all organs and tissues of the body. In fact, the diabetic state is one of the major contributors to deaths caused by myocardial infarction, renal failure, cerebrovascular disorders, arteriosclerotic heart disease and systemic infections.

The conditions regarding hyperglycaemia and glycoseria in the diabetic disorder can be corrected by changing the diet, controlling body weight and regulating physical activity. In some diabetics, especially those suffering from NIDDM, the conditions regarding hyperglycemia and glycoseria can be managed by oral administration of anti-hyperglycemic agents such as derivatives of sulfonylureas, sulfonamides, biguamides and other compounds. However, for diabetics suffering from IDDM and advanced NIDDM, treatment has focused on management

of insulin of foreign origin.

Insulin is a polypeptide that is formed in the islets of Langerhans

in the pancreas. The insulin molecule is synthesized to and

begin as a simple polypeptide chain, but it is further processed so that in its active form it consists of two amino acid chains linked together by two cysteine disulphide bonds. One of the two chains is folded back on itself as a result of a third disulfide bond. The entire molecule has a molecular weight of 5.734 and is dependent on the disulfide bonds to retain its biologically active conformation.

While insulin of foreign origin has previously mainly come from bulls and pigs, it has recently been obtained in "human" form as a result of recombinant DNA technology. Availability of "human" insulin that comes from recombinant sources has proven to be very beneficial for those diabetics who are hypersensitive to insulin that comes from animal sources.

Nevertheless, the biggest problem when it comes to insulin therapy is not associated with the source of the insulin, but rather the way in which it is introduced into the body. The most common way to administer insulin is the subcutaneous injection. This way is uncomfortable, painful and can even worsen the pathology of the disorder. Subcutaneous injection of insulin gives rise to relatively high insulin levels in peripheral tissues, and relatively low levels circulating through the liver, the main seat of endogenous insulin activity. High insulin levels in peripheral tissue are associated with blood vessel pathology (eg, blood vessel constriction and changes in flow) and pathological effects on associated peripheral tissue, eg, diabetic retinopathy. The "over-swimming" effect of subcutaneously administered insulin on peripheral circulatory tissue ultimately reduces the amount of insulin circulating to the liver, which in turn results in the need for increased doses to achieve the desired metabolic effect. Of

for these various reasons, an alternative to injection as a method of administering insulin has long been sought.

As an alternative to injecting insulin, some researchers have directed their efforts towards intrarectal administration of insulin by means of suppositories. U.S. Patent No. 2,373,625 issued to Brahn, mentions insulin suppositories containing weak organic acids such as lactic acid or citric acid together with a surfactant.

Also known in the field are insulin suppositories where a number of ingredients are used such as saponin, corn oil, polyoxyethylene-9-lauryl alcohol and polyoxyethylene lauryl ether. Insulin has also been encapsulated in acrylic acid-base water-soluble gels and in soft gel capsules containing surfactants. Although the procedure of intrarectal administration of insulin seems promising, the results of bioavailability tests have been inconclusive and the procedure is inconvenient.

Ishida et al., Chem. Pharm. Bull., 29, 810 mentions a procedure whereby insulin can be administered via the mucous membranes of the cheek in the mouth. Insulin was mixed with a cocoa fat base and a surfactant before administration to the buccal mucosa. Experiments with dogs show only poor bioavailability of insulin administered in this way.

Other researchers have directed their efforts to the intrapulmonary administration of insulin. Wigley, et al., Diabetes, 20, 552 (1971) discusses administration by nebulizer of insulin at 500 U/ml in particles 2 micrometers in diameter. A hypoglycaemic response was observed after 30 U/kg body weight indicating a total bioavailability of approx. 7 to 16%. Yoshida, et al., J. Pharm. Pollock. 68, 670 (1979) mentions an intrapulmonary introduction of insulin together with lactose and acetylglycerin monostearate dissolved in fluoroethane. The aerosol produced a hypoglycaemic response when administered at an amount of 2.5 U/kg body weight to rabbits.

Another procedure for administering insulin that has shown promise is the intranasal administration of insulin. Hirai, et al., Int. J. Pharm., 9, 165 (1981) discusses the intranasal administration of insulin together with a surfactant such as sodium glycocholate solution. Nagai, et al., Journal of Controlled Release, 1, 15 (1984) reports high bioavailability of insulin when administered intranasally to dogs. In this procedure, crystalline insulin was dissolved in a 0.1 N HCL solution to which a surfactant was added. The pH of the solution was then adjusted to 7.4 by adding 0.01 N NaHCL solution and freeze-dried. The solution was then mixed with crystalline cellulose prior to its nasal administration.

In the search for alternative procedures for the administration of insulin, it is the oral administration of insulin that has received the most attention. A procedure for the oral administration of insulin would be highly desirable for reasons of safety, convenience and comfort and would avoid many of the shortcomings that procedures involving the injection of insulin suffer from. Despite the apparent desirability of methods of oral delivery of insulin, two significant difficulties have limited the success of attempts to develop oral insulin therapy.

The first significant difficulty in preparing oral insulin therapy is that insulin polypeptide is inactivated in the gastrointestinal tract by enzymes such as trypsin, chymotrypsin and other lytic enzymes. The insulin polypeptide is relatively simple and with its two disulphide bonds it is easily degraded under the harsh conditions in the stomach and gastrointestinal tract.

The second difficulty with oral administration of insulin is that although the polypeptide avoids degradation in the stomach and gastrointestinal tract, it is poorly and unreliable absorbed through the membrane of the gastrointestinal tract. Due to its poor bioavailability, large oral doses must be given to ensure a hypoglycemic effect. Because unreliable amounts of insulin are made available, the advantages of an oral administration procedure may be outweighed by the fact that under- or overdoses of insulin may be a greater health risk than no insulin at all.

A number of approaches have been used in attempts to overcome the difficulties associated with oral insulin administration. Some of these approaches include attempts to either inactivate the lytic gastrointestinal enzymes responsible for inactivating insulin function or to provide insulin analogues that are resistant to inactivation by such enzymes. Sichiri, et al., To-Nyo-Byo (Japanese Diabetes Publication), 18:619, 1975, discloses an insulin analog (beta-naphthyl-azo-Polystearyl insulin) which when administered orally to rabbits at 150 IU/kg body weight gave a hypoglycemic response. Other researchers have attempted to make alkyl compounds with insulin by adding triethylamine HCL as well as a surfactant.

Teng, oral presentation at the American Diabetic Society

Meeting, May, 1983, San Antonio, Texas.

Attempts to prevent the breakdown of insulin by inactivating lytic enzymes in the gastrointestinal tract have met with some success. Danforth, et al., Endocrinology, 65, 118 (1959) discloses an insulin preparation administered orally together with isopropyl fluorophosphate (an inhibitor of trypsin and chymotrypsin) and indole-3-acetate (an inhibitor of the enzymes known as "insulinase " found in the liver). The preparation was found to produce a hypoglycaemic effect when administered orally to rats. Other researchers report weak (equivalent to 3% bioavailability of insulin) hypoglycemic response after oral administration of insulin together with agents that inactivate the pancreas (Laskowski, et al., Science, 127, 1115 (1958)). Other researchers have found that oral administration of insulin results in low bioactivity as a result of inactivation of insulin in the stomach by pepsin and due to poor absorption through the intestinal membrane (Crane, et al., Diabetes, 17, 625 (1968)).

Researchers have also attempted to increase the bioavailability of orally administered insulin by co-administering the insulin with surfactants. The use of surfactants such as polyethylene glycol-1000 monoacetyl ether and sodium lauryl sulfate together with triethylamine HCL has shown the ability to increase insulin bioavailability to some extent: although 35 units of insulin administered orally had an effect on blood glucose levels equivalent to only 4 units administered intravenously ( Touitou, et al., J. Pharm. Pharmacol., 32, 108 (1980)).

Efforts have also been directed towards oral administration of insulin together with an emulsification system. A group of researchers (Sichiri, et al., Acta Diabet. Lat., 15, 175 (1978)) report that they observed a hypoglycemic effect when administering a water/oil/water emulsion system with added insulin to rats. A dose of 250 IU/kg body weight was reported to produce an effect equivalent to a 10 IU/kg dose administered intramuscularly. Insulin has also been administered by means of other emulsion systems using fat-soluble vitamins although the hypoglycaemic effect in animals such as dogs has been weak and has not responded to the dose.

Some of the most promising attempts to increase the bioavailability of orally administered insulin lie in the attempt to microencapsulate the insulin material. Insulin has been microencapsulated in the acrylic acid ester (Sichiri, et al., Acta. Diabet. Lat., 15, 175

(1978)) and the use of high molecular weight polymers have also been discussed. Other researchers have microencapsulated insulin in liposomes of different lipid composition. Insulin containing liposomes have been formulated with compounds such as phosphoidyl choline, cholesterol and stearylamine; dimyristoyl phosphatidylcholine; dimyristoyl phosphatidylcholine and cholesterol as well as with lecithin and cholesterol and other materials.

Dobre, et al., Rev. Roumanian Med.- Encdocrinol 22, 253

(1984) discuss methods for encapsulating insulin in liposomes for oral administration. Various materials for forming liposomes are mentioned including: phosphatidylcholine from egg yolk, cholesterol, stearylamine and dipalmitoylphosphatidylcholine. However, even liposome coating fails to completely protect the contained insulin from the degrading effects of hydrochloric acid found in the stomach and enzymes such as pepsin, trypsin and chymotrypsin found in the gastrointestinal tract. Yoshida, et al., EPA 140,085 discloses insulin containing lipid vesicle preparations containing lipid vesicles consisting of an internal aqueous phase in the form of an aqueous solution or suspension of phospholipid. The present invention provides compositions adapted to administer pharmacologically effective amounts of protein compounds when ingested orally. It has surprisingly been found that pharmaceuticals which tend to be broken down in the gastrointestinal tract and lack the ability to be effectively absorbed into the body through this can be coated in such a way as to ensure their absorption into the body in pharmacologically effective amounts. The improved compositions comprise: (a) a pharmaceutical agent; and (b) lipid coating material; and may include (c) enteric coating material. The pharmaceutical agent coats with an inner coating consisting of the lipid coating material and with an outer coating consisting of the enteric coating material. The mixture may comprise or may be co-administered with lipolytic enzymes. The mixtures may additionally comprise water-soluble coating materials, binding materials, surface-active materials, emulsifiers, stabilizers, antimicrobial agents and enzyme inhibitors. In another embodiment of the present invention, the improved composition comprises: (a) a pharmaceutical agent; (b) lipoprotein materials; and (c) a lipid coating, and may comprise (d) a water-soluble coating and (e) a gastric juice resistant material. The pharmaceutical agent is bound to lipoproteins and the resulting product can be coated with a water soluble coating and then coated with a lipid inner coating which can be coated with an enteric outer coating. The mixture may comprise or may be co-administered with lipolytic enzymes. The mixtures may additionally include binders, surface-active materials, emulsifiers, stabilizers, antimicrobial agents and enzyme inhibitors. In one embodiment of the invention, insulin is mixed with a mixture consisting of emulsifier/dissolution aid, surfactant, enzyme inhibitor, anti-foaming agent and anti-microbial agent. The mixture is then coated with a lipid coating comprising the polyethylene glycol fatty acid ester. The material is then either further coated with an enteric coating and either filled into hard gel capsules or tablets or filled into a capsule or tablet which is then coated. Either tablet or capsule weighs 250 mg and contains 16 international units of crystalline insulin extracted from pigs. In another embodiment of that invention, insulin is mixed with a mixture consisting of at least one: lipid; amino acid; binder; enzyme inhibitor; anti-foaming agent and water-soluble material. The mixture is then coated with a lipid coating consisting of triglycerides, phospholipids and cholesterol. The resulting product is then filled into a gel capsule and is enteric coated. In a further design of the present invention, it comprises oral insulin preparation, which has a size of approx. 2.00 mm. Preferably, citric acid particles are coated with sodium bicarbonate to form a particle which is then coated with a lipid. The resulting particle is coated with an oral insulin preparation and then with an enteric solution. In yet another embodiment, insulin particles coated with a lipid coating layer and then enteric-coated are filled in a capsule with enteric-coated particles of lipase and bile salt. Preferably, sodium bicarbonate and citric acid are also added to the capsule. The apparatus for making the mixture consists of a vessel which has a chamber. At the bottom of the chamber there is a perforated rotating disc, equipment for air flow and a chopping knife. Nozzles are located above the bottom of the chamber. To make the mixture, particles are placed in the bottom of the chamber and chopped with the chopping knife. The air flow equipment causes the particles to be suspended inside the chamber. A pharmaceutical agent is sprayed through the nozzles so that the agent binds to the particles. After the agent has bound to the particles, the particles it has bound to are then coated with a lipid coating that is sprayed through the nozzles. After the lipid coating is applied, an enteric coating can be applied. Figure 1 shows, in section, a schematic representation of a "jet mill". Figure 2 shows a section view of the "jet mill" in Figure 1. Figure 3 shows a section view of a schematic view of a Spir-A-Flow. Figure 4 is a chromatograph of the mixture in Example 1. Figure 5 is a photomicrograph of the mixture in Example 1. Figure 6 is a photomicrograph of the mixture in Example 1. Figure 7 is a comparison between the venous serum sugar level in two pigs, one has received the mixture from example 1 and one has received aktrapid. The venous serum sugar level of the pig that has received example 1 is marked "o" and the venous serum sugar level of the pig that has received actrapid is marked "x". The X-axis is in minutes and the Y-axis is in millimeasures of serum glucose. Figure 8 is a comparison between Tong-Shin insulin, the oral insulin from Example 1 and Novo insulin, after the test in test 1-D. Figure 9 is a comparison between the oral insulin from example one and regular injected insulin after test 1-E. Figure 10 is a comparison between the oral insulin from example 1 and oral insulin from example 2 after test 2-B. Figure 11 is a comparison between the oral insulin from Example 1 and the oral insulin from Example 2 after 2-B. Figure 12 shows the effect of Example 6 on blood sugar levels. Figure 13 is a comparison of injectable insulin compared to Example 6. Figure 14 is a comparison of injectable insulin compared to Example 6. Figure 15 is a comparison of injectable insulin compared to Example 6. Figure 16 shows the effect of Example 6 on blood sugar levels . Figure 17 shows the effect of Example 6 on blood sugar levels. Figure 18 shows the effect of Example 6 on blood sugar levels. Figure 19 is a comparison between injectable insulin and example 6. Figure 20 is a comparison between injectable insulin and example 6. Figure 21 is a comparison between injectable insulin and example 6. Figure 22 is a comparison between oral insulin from example 7 and regular injectable insulin. Figure 23 schematically shows a section of a CF granulator. Figure 24 is a schematic depiction of an oral insulin granule from example 12. Figure 25 shows the insulin levels in the blood of dogs that have received insulin preparations from example 12. Figure 26 shows the effect of the oral insulin granules from example 12 on dogs' blood sugar levels. Figure 27 shows the effect of the oral insulin from Example 12 on dogs' plasma sugar levels. Figure 28 shows the effect on blood sugar of the oral insulin preparation from example 11 in relation to the oral insulin preparation from example 12. Figure 29 shows the effect on blood sugar of the oral insulin preparation from example 13. The present invention provides a mixture adapted for oral administration of a selected protein compound material in biologically active form. As previously discussed, many drugs, medications and treatments cannot be administered orally because they are broken down or not well absorbed in the stomach and/or gastrointestinal tract. Examples of these medicines include, among others: insulin - for the treatment of diabetes; urokinase - for the treatment of thrombosis; Factor VIII - for the treatment of hemophilia; leuprolide - for the treatment of prostate cancer; gangliosides - to improve neurotransmission; vincristine - for the treatment of cancer; belomyein - for the treatment of cancer; adriamycin - for the treatment of cancer; Lidocaine - for the treatment of cardiac arrhythmia; and cephalosporins - to treat infection. Other drugs and medications are poorly absorbed when administered orally, for example, gentamicin, erythromycin, bretylium tosylate, setiedil, cyclandelate, and chloramphenicol. For most patients, oral administration of these drugs would be more appropriate and for some more therapeutic. The present invention provides a mixture which allows protein compounds which previously had to be administered parenterally to be administered orally in pharmacologically effective amounts. With this purpose, in one embodiment of the invention, the mixture comprises particles consisting mainly of solid emulsifiers and surfactants where a selected biologically active protein material is bound to the surface of the particles with a binder. A lipid coating is then applied over the particles and protein material. The particles may have an average diameter of approx. 1 micron to approx. 1/2 millimetre, but preferably the particles have an average diameter of approx. 1 to around 100 microns. The lipid coating is preferably applied to the particles and protein material in a thickness of approx. 0.05 to 1.0 micron. The lipid coating can be coated with a gastric juice-resistant coating layer. In another embodiment of the invention, the protein material is bound to lipoprotein particles. The particles may have an average diameter of 1 micron to 1/2 millimeter, but preferably the particles have an average diameter of 1 to 100 microns. The particles are then coated with a water-soluble material and the resulting product is coated with a lipid coating. The lipid coating preferably has a thickness of approx. 0.1 to approx. 0.3 micron. The resulting product can be filled in a gel capsule and can then be enteric coated to a thickness of approx. 0.1 micron to about 0.3 micron. In another embodiment, the protein material is applied to the grains. The granules are formed by applying sodium bicarbonate and, if desired, a small amount of starch, to particles of citric acid (preferably 100 to 150 grams in size). A preparation comprising protein material is then applied to the grain. The resulting product is then enteric coated. The resulting grains preferably have a size of less than 2.0 millimeters. Physiologically, a grain size of less than 2.0 millimeters can be rapidly emptied into the duodenum. By encapsulating the grains in small hard gel capsules, as opposed to larger gel capsules, absorption in the intestines is also increased. The use of sodium bicarbonate and citric acid, either when the grains are made or used in preparation as a powder, accelerates a rapid bursting and opening of the capsules or grains and thus releases the protein material in the duodenum. Of course, other compounds can be used to accelerate this bursting effect, such as sodium citrate and other bicarbonates, such as potassium bicarbonate. The gastric juice-resistant coating ensures that the mixture passes down the intestinal tract without being broken down in the stomach. In the intestinal tract, the coating dissolves. The lipid coating ensures that the mixture passes into the user's lymphatic system. To this end, the lipid coating gives the mixture an asinity towards the tufts (the end openings of the lymphatic system in the intestinal membrane). If the pharmaceutical agent is bound to lipoprotein particles, these particles increase the agent's absorption into the tissue. The lipoprotein has been found to easily penetrate the cell membrane and therefore provides a carrier substance to bring the agent into the user's tissues. It is believed at this point that when the pharmaceutical agent is insulin, lipolytic enzymes should also be administered. The lipolytic enzymes can be combined with the mixture or given in a separate tablet or capsule. The lipolytic enzymes increase the absorption of the lipid-coated material in the tufts. The preferred ingredients for the mixture according to the present invention are now presented.

Protein material.

The biologically active protein materials or pharmaceutical agents useful in the present invention may include, but are not limited to, those agents which tend to be degraded in the gastrointestinal tract or are poorly absorbed into the body from the gastrointestinal tract. These agents include insulin, either from an animal or recombinant source, as well as other pharmaceutical agents which include, among others, urokinase, Factor VIII, leuprolide, gangliosides, vincristine, belomyein, lidocaine, cephalosporins, gentamicin, bretylium tosylate, seteidil, cyclandelate, erythromycin, chloramphenicol, adriamycin and streptokinase.

Lipid coating materials.

Lipid coating materials useful in the invention include the cyclomicron-like materials that are occasionally attracted to the tufts of the small intestine. Such materials include, but are not limited to, the polyethylene glycol fatty acid ester, glycero-phosphatides, foot fatidyl phosphates, egg yolk lecithin, oleic acid, stearic acid, palmitate, cholesterol, mono, di- and triglycerides, cholesterol esters, egg yolk lecithin containing 5 to 20% phosphatidic acid, linoleic acid , linolenic acid, lauric acid, phosphatidyl phosphate, glycerin, soybean oil, sesame oil and tromethane. These lipid coating materials, after being applied to the surface of the pharmaceutical agent, will not only seal the pharmaceutical agent, but also increase the "affinity" and "absorption" of such drug particles to and towards the tufts, that is, the end openings of the lymphatic vessels in the intestinal membrane, such that the agent is absorbed in the lymphatic system.

A lipid coating mixture that has been found to work satisfactorily is the following: 80 to 90% lipid can be in the form of mono-, di- and tri-glycerides; 6 to 9% phospholipids;

2% cholesterol esters and 1% free cholesterol; and

less than 1% free fatty acids (NEFA).

About. 2% low molecular weight protein/amino acid can also be added. The cholesterol esters are: 30% as cholesterol-linoleic acid ester;

20% as cholesterol-oleic ester; and

22% as cholesterol-palmitic acid ester.

Gastric acid-resistant cover materials.

enteric coating materials useful in the present invention include those coating materials which resist degradation in the stomach, but which will decompose in the environment in the intestinal tract and release the coated material. Such enteric coating materials include, but are not limited to, the following ingredients, either individually or in admixture: hydroxypropylmethylcellulose phthalate, polyethylene glycol-6000, and shellac, and they may be dissolved in solvents including chloromethane, ethanol, and water, cellulose phthalate or polyvinyl acetate phthalate.

A preferred enteric coating material for coating insulin comprises 4.5 parts by weight hydroxymethylcellulose to 0.5 parts shellac, to 0.5 parts polyethylene glycol-6000. This material is dissolved in 47.3 parts dichloromethane and 37.8 parts ethanol. The gastric juice-resistant coating material is then diluted with water to the optimum concentration and applied to the preparations according to the invention.

Lipoprotein materials.

The lipoprotein materials useful in the present invention include those lipids and proteins which can combine to form lipoproteins with a low specific gravity. Preferably, the low specific gravity lipoproteins mimic the profile of low specific gravity lipoproteins in humans.

The lipid components include, but are not limited to, cholesterol, oleic acid, stearic acid, sodium lauryl sulfate, lecithins, sodium lauryl sulfate with lecithins, phosphatides, palmitic acid, and phosphatide phosphate choline.

The protein components include, but are not limited to, all amino acids or low molecular weight proteins. The most preferred amino acids include arginine, lysine, histidine and aspartic acid, and the most preferred proteins include albumin and globulin.

Preferably, the lipids are mixed with low molecular weight protein or amino acids in a molecular weight ratio between 1:1 and 3:1. In a preferred embodiment, the lipids and proteins are mixed in a 1:1 ratio.

A lipid mixture comprising cholesterol esters (46%), free cholesterol (14%), phospholipids (25%), fat (14%) and NEFA (FFA) around 1% has been found to work satisfactorily when mixed with 20 to 25% by weight of amino acids.

Water-soluble coating materials.

Water-soluble coating materials useful in the present invention include those compounds which provide a hydrophilic coating, including but not limited to hydroxypropylmethylcellulose phthalate, polyethylene glycol-6000, hydroxypropylmethylcellulose, hydroxypropylcellulose, crystalline cellulose and shellac, dissolved in dichloromethane, ethanol and water, cellulose phthalate and polyvinyl acetate- phthalate. If a protein material is bound to the lipoproteins and coated with a water-soluble coating, they are preferably coated with a coating having a thickness of approx. 0.05 to approx. 0.5 microns.

Binding materials.

Binding materials useful in the present invention are those compounds which bind a peptide to a lipid or lipoprotein. These include, but are not limited to, microcrystalline cellulose, methyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methyl hydroxypropyl cellulose, gelatin, povidone, and polyethylene glycol fatty acid ester groups.

Lipolytic enzymes.

Lipolytic enzymes useful in the present invention are those enzymes which aid the absorption of lipids in the intestinal tract. These enzymes include, among others, lipase, including pancreatic lipase, anylase, protease, and bile salts. Absorption in the intestines of fatty acids and lipids is increased and assisted by pancreatic lipase in the presence of bile salts. Accordingly, when the protein material, when coated with lipoprotein pancreatic lipase in the presence of a bile salt, the absorption of the protein material will increase. If necessary, the lipolytic enzymes can be mixed with preparations or administered as a separate tablet or capsule. A lipolytic enzyme mixture that has been found to work satisfactorily when the pharmaceutical agent is insulin includes: pancreatic lipase, amylase, protease and bile salts.

Fillers.

If the protein material is bound to a lipoprotein, a filler can be used. The filler may or may not be a physical binder. The filler may include, but is not limited to, gum arabic, crystalline cellulose, hydroxypropyl cellulose and hydroxypropyl methyl cellulose.

Surfactant materials.

Surfactant materials useful in the present invention include those compounds that rapidly improve absorption in the intestinal tract and include, but are not limited to, cationic, anionic or nonionic surfactants such as sodium lauryl sulfate, stearylamine, polyglycerin fatty acid ester, polyethylene alkyl ether, polyoxyethylene alkyl vinyl -ether, fatty acid monoglycerides, sorbitan fatty acid ester, polyoxyethylene fatty acid amines and polyethylene glycol fatty acid ester.

Emulsifiers.

Emulsifiers useful in the present invention include those compounds that create an emulsion in vivo, and include, but are not limited to, the materials and mixture of materials, such as cholesterol, stearic acid, sodium stearate, palmitic acid, sodium palmitate, oleic acid, sodium oleate glycerin monostearate, polyethylene 50 stearate, polyoxy 40 stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, propylene glycol diacetate, and propylene glycol monostearate. The emulsifier may also include the chemical materials and constituents that make it the equivalent of "low specific gravity glypoproteins" found in human plasma, such as, arginine HCL, gum arabic, cholesterol, cholesterol esters, phospholipids and fatty acids.

Stabilizers and enzyme inhibitors.

Stabilizers and enzyme inhibitors which are useful in the present invention are those compounds which are capable of inactivating or preventing a degrading effect of mammalian enzymes such as peptidases, proteases, phosphorylases, glycation-insulin-transyhydrogenase and insulin-degrading enzyme, i.e. insulinase. The stabilizers include, but are not limited to, stearylamine, stearyl alcohol, citric acid, lactic acid, triethylamine HCL pyrophosphate, triethanolamine, ovomucoid, ethylenediaminetetraacetate, iodoacetamide, phenylhydrazine, hydroxylamine, and 8-hydroquinoline.

Antimicrobial agents.

Antimicrobial agents useful in the present invention are those compounds capable of preventing microbial contamination and degradation of the pharmaceutical compound and its accompanying binders, coatings and additives. When an antimicrobial agent is selected, the microbial action of the agent must be balanced against its compatibility with other components of the pharmaceutical preparation as well as its accompanying toxicity to the organism treated with it. Antimicrobial agents that generally meet these requirements include, but are not limited to, methylparaben, ethylparaben, propylparaben, butylparaben, phenol, dehydroacetic acid, phenylethyl alcohol, sodium benzoate, sorbic acid, thymol, thimerosal, sodium dehydroacetate, benzyl alcohol, butylparaben, cresol, p-chloro-m -cresol, chlorobutanol, phenyl mercuric acetate, phenyl mercuric borate, phenyl mercuric nitrate and benzyl alcohol chloride.

The invention will be more fully understood by reference to the following examples.

Example 1.

In this example, a mixture adapted for oral administration of insulin is prepared according to the invention. Particles that have an average diameter of approx. 50 micron was prepared from a mixture of cholesterol, sodium lauryl sulphate and methyl and propyl parabens (pH 6) as antimicrobial preservatives. The particles were prepared using a Jet Mill available from Freund International Ltd., Tokyo, Japan.

Figures 1 and 2 schematically show the Jet Mill 10. The particles to be processed are fed into the intake chamber 11 by a vibrating feeder. The particles in the intake chamber are fed into a mill chamber 12 by means of an air flow through an air nozzle 13. An opposing air flow is applied through air nozzle 14 which goes against the flow of air which carries the particles in suspension. The air currents are shown by arrows. This results in a grinding action of the particles within the circular mill chamber 12. The air flow through the nozzles is controlled to adjust the grinding action within the chamber. The particles are ground to the desired diameter size and then fed to an air discharge port 15 in the middle of the mill chamber. A separator 6 directs the ground particles so that they are stored in a collection flask 7 while the air is sucked out through the air outlet pipe opening 8.

Crystalline monocomponent porcine insulin (obtained from

Novo, Bagarvaerd, Denmark) was then bound to the particles using a solution containing 8% by weight hydroxypropyl cellulose and 0.005M sodium lauryl sulfate, 0.005M triethylamine HCL and 0.005M citric acid. The particles with insulin bound to their surface were then coated with a solution consisting of 6% by weight polyethylene glycol monostearate (EMANON-3199 obtained from KAO, Tokyo, Japan) so that a lipid coating was formed approx. 0.1 micron thick. An enteric coating solution was then prepared by dissolving 4.5 w/w% hydroxymethylcellulose phthalate (HPC-55), 0.5 w/w% shellac (Shin-Etsu Chemical, Japan) and 0.5 w/w% polyethylene glycol- 6000 (Shin-Wha Yakuhin, Japan) in a solution containing 47.3 parts by volume of dichloromethane and 37.8 parts by volume of ethanol. The solution is then diluted with water to an optimal concentration so that the lipid-coated particles are then coated with the gastric juice-resistant layer to a thickness of approx. 0.1 micron.

250 mg of the enteric coated particles are then filled by hand into hard gelatin capsules.

The mixture was made using a modified

"Freund" Spir-A-Flow, model SFC-Mini-S 220 V, 60HZ, 3P (available from Freund International, Tokyo, Japan). The standard Spir-A-Flow was modified as follows. The mesh holes in the rotor were found to be too large. Accordingly, additional maskings were added. The masks were also oriented so that the particles would not fall through.

The Spir-A-Flow was also modified so that the air being pumped was dried and had all oil droplets removed. The air was dried to 20% absolute humidity. The airflow pressure was also increased to 920 liters per minute and the airflow was oriented so that it was blown up through the bottom.

Also, instead of one air intake opening at the bottom, four openings were used. The openings were oriented in the bottom at 0°, 90°, 180° and 270°.

The bag filters and the pulsating filters were also shortened to half their length.

Also, instead of one nozzle placed at the bottom of the tub, three nozzles were placed at the bottom of the tub. A fourth overdraft nozzle was also placed on top of the vessel. The bottom nozzles were used for binding and the top nozzle for covering.

Figure 3 shows a Spir-A-Flow device 20. Briefly, the Spir-A-Flow device 20 works as follows. The particles are placed on the rotating disc 22. The rotating disc 22 comprises the mesh holes 31. A cutter 29 and stirrer 21 cooperate with the disc 22 to form particles of a given size. The air flow is then switched on and the particles are suspended in the vessel 26. Two types of air flow are formed, air flow in a gap and flowing air. The air stream in the gap enters through opening 33 while its flowing air enters through opening 32. The suspended particles can then be coated with a binder solution sprayed in from the low nozzles 27. Although only a low nozzle 27 is drawn in the modified Spir-A-Flow, three low spray nozzles 27 were used. When the particles are coated.

they fall back onto disc 22. When all the particles are coated with binder, the air flow is increased and the coated particles can be coated again, etc. As discussed here later, the particles can also be coated with solution sprayed on from the upper nozzle 28.

Due to the repeated movement with binding and coating of the powdered particles, the particles are well mixed, moistened, kneaded and mixed very efficiently and are transformed into high quality granules very quickly. A hot air flow through the perforated areas on the disc dries the grains effectively and efficiently. If dusty material is formed and lifted to the top, this dust is collected in the bag filters 24. Cleaning nozzles with air flow 25 are used to blow the dusty material collected in the filters back to the disc 22 at the bottom as needed.

The rotating disk 22 and the chopper 29 work together to form particles that have a round and smooth surface, with great mechanical strength, high bulk density and little dispersion of the particle size. To make the mixture in this example, the solid emulsion solvents, surfactants, antimicrobial preservatives were poured into the vessel through an intake. While the powdery raw materials were being filled into the vat, gentle currents of air were blown upwards, through a perforated part of the rotating disk and a surrounding gap between the rotating disk and the inner wall of the chamber (vessel). The air flow prevents the materials from falling down through the slit and perforated parts.

The rotating disc 22, agitator 21 and chopper 29 were started after the raw materials were charged in. To make the mixture of Example 1, the rotor (and thus the perforated disc) was rotated at a speed of 300 revolutions per minute (r.p.m.), the chopper at 2,000 r.p.m., and the stirrer rotated at 500 r.p.m. Air currents at a temperature of 32 to 35°C pass through the openings in the rotor ("flowing air") at a flow rate of 8 to 10 liters per minute, and through the gap between the rotor and the wall ("gap air") at 7 to 8 L/min. The collected particles were dried for 30 minutes.

Crystalline insulin (Novo Monocomponent Porchin Crystalline Insulin, Batch No. 833115, Lot No. 69195-01) was carefully dissolved in a solution containing hydroxypropyl cellulose, sodium lauryl sulfate, citric acid, and triethylamine HCL, pH 2.0.

While the particles were blown upwards inside the wall by the cover and the binding apparatus by "gap air" and "flowing air", the internal solution was sprayed as a fine mist through two spray nozzles with a spray air pressure of 3.0 kg per M2 and at an air flow rate of 10 L/min.

The insulin-coated solid emulsification particles were then coated with a lipid coating consisting of a solution of the polyethylene glycol fatty acid ester (KAO SEKKEN, Japan: EMANON 3199, Lot No. 12744Y) with a spray air flow rate of 10 L/min and a spray air pressure of 3.0 kg/M2.

The above coated insulin-coated solid emulsifying particles were then filled into hard gel capsules, these capsules were then coated with an enteric-resistant coating.

Each resulting 350 Mg capsule contained:

(350 mg Cap.)

Cholesterol (Fuluka, A.G.; Switzerland;

Lot no. 24657-784) -6.76 x 10"<4>M

Sodium lauryl sulfate (Yakuri, Kagaku,

Japan; S316-06) -1.377 x 10-<4>M Triethylamine HCL (Wako Sei-Yaku, Japan) -4.089 x 10"6M Citric acid (Yakuri Kagaku, Japan;

KA9105) -5.965 x 10"<6>M

Hydroxypropyl cellulose -L (Shin-Wa Yakuhin,

Japan; 191184) -1.234 x 10"<2>G

Polyethylene glycol fatty acid esters

(KAO Sekken, Japan; EMANON 3199;

12744Y) -1.543 x 10"<2>G

Insulin (NOVO) -8 units

Citric acid is used as an inhibitor for insulin-inactivating enzymes and as an insulin stabilizer and for pH adjustment.

Triethylamine HCL is used as an insulin stabilizer and as an anti-foaming agent.

Test 1-A

In this test, the oral insulin mixture from Example 1 was chemically compared to a sample of Actripide intravenous insulin solution (Novo). A capsule of the oral insulin preparation containing 8 international units (I.U.) Novo monocomponent crystalline insulin prepared from porcine, prepared in accordance with Example 1 was dissolved, by "sonification" using an "ultrasonicer" for five minutes in water. Half (1/2) of the solution — containing 4 I.U. of the oral insulin preparation

- was analyzed using reverse phase chromatography. 4 I.U. of the Novo actrapid insulin was also analyzed using reverse phase chromatography. The results indicated that both samples contain the same amount of chemically equivalent insulin. (See figure 4.)

Test 1- B

In this test, photomicrographs using a scanning electron microscope of the oral insulin preparation from Example 1 were taken. The gastric juice-resistant and lipid coatings on the particles were removed using a solvent which was 47.3 parts by volume of dichloromethane and 37.8 parts by volume of ethanol in water. Photomicrographs of the particles at 500X magnification showed relatively small particles (about 5 to 10 microns in average diameter) of insulin and hydroxypropyl cellulose binding materials attached to relatively large particles (about 50 microns in average diameter) of cholesterol and sodium lauryl sulfate materials. (see figure 5). At 3000 times original size, rod-shaped insulin crystals become visible impregnated with the hydroxypropyl cellulose particles. (See figure 6).

Test 1- C

In this test, the oral insulin preparation from the example was used

1 compared to actripide intravenous insulin solution (Novo) in a bioassay. 2 Yorkshire pigs fasted overnight and one pig received 8 I.U. actripide insulin intravenously. The second pig received an intravenous infusion over a period of 15 minutes of 8 I.U. of the oral insulin from example 1 which was dissolved in 50 ml of water. Blood was drawn from the pair of pigs and serum glucose levels were measured. The venous serum sugar level in the two pigs decreased to approximately the same extent, although the duration of the hypoglycemic effect for the preparation from Example 1 was somewhat longer than that for actripide insulin. (See figure 7).

Test 1-D

In this test, the oral insulin preparation from the example was used

1 compared with Novo monocomponent crystalline insulin prepared from porcine and a regular injectable insulin-40 (Tong-Shin Pharmaceutical Co.) in intravenous bioassay with rabbits. Three white rabbits each weighing 1.8 kg were fasted overnight and treated by intravenous infusion over 10 minutes of 4 I.U. of either Novo insulin, Tong-Shin insulin-40,

or an emulsion consisting of the oral insulin preparation from Example 1. A venous blood sample was collected from the ear vein of each of the rabbits 30 minutes before the insulin infusion and at time points 0, 15, 30, 45, 60, 75, 90 and 105 minutes after completion of the infusions.

Each of the three forms of insulin showed approximately the same hypoglycaemic effect and the onset of the hypoglycaemic effect occurred within 30 minutes after the start of the infusion. The time of peak hypoglycemic effect of Novo-insulin was determined to be within 30 minutes after completion of the infusion, while for Tong-Shin-insulin-40, it was 60 minutes. The emulsion with the oral insulin preparation of Example 1 peaked for hypoglycemic action between Novo and Tong-Shin insulin. (See figure 8).

Test 1- E

In this test, the oral insulin preparation from the example was used

1 administered orally to two healthy male volunteers. Administration of the oral insulin induced hypoglycemic effects comparable to those caused by subcutaneous injection of an equivalent amount of injectable insulin (Tong-Shin).

Two healthy male volunteers, aged 37 and 38, weighing 52 and 54 kg, participated in the study. Each subject received a 16 IU dose of the oral insulin preparation of Example 1 during the course of the study. Along with the oral insulin preparation of Example 1, each participant received two capsules of lipolytic enzymes. Each capsule consisted of:

per 350 mg capsules, using corn starch as "filler".

The enzyme capsules were made as follows: Pancreatic lipase was coated with a 0.1 to 0.3 micron thick coating of hydroxypropyl cellulose and enteric coated with the coating from Example 1. Amylase and protease were then added. The resulting preparation was then coated with 0.1 micron hydroxypropyl cellulose. Bile salts coated with 0.1 micron hydroxypropyl cellulose were then added to a hard gel capsule along with the resulting pancreatic lipase product.

At 7:30 a.m. on the test day, both participants consumed a standard breakfast consisting of two fried eggs, two slices of bacon, two slices of toasted dark bread, a tablespoon of butter, and a cup of black coffee. The insulin preparation from Example 1 was administered orally at 10:00 in the morning to both subjects. The oral insulin was administered in capsules containing 16 I.U. Novo crystalline monocomponent insulin from porcine sources.

At 12:00 a.m., both subjects consumed a standard lunch consisting of 180 gr. fish, a green salad with oil and vinegar dressing, a piece of dark bread and black coffee.

Blood samples were taken from the objects at 10:00, 13:00 and

16:00 and was analyzed with regard to blood sugar levels. The tests indicated that the oral insulin preparation produced a strong hyperglycemic effect in both subjects after three hours, but that after six hours the blood sugar levels had returned to their pre-treatment levels.

Blood glucose levels had dropped dramatically for both subjects three hours after administration of the insulin. About. At 10:45 the first subject began to complain of blurred vision, inability to concentrate and nausea. 11:20, the same subject began to complain of cramping and trembling in arms and legs and cold sweat. The other object also complained about approximately

Example 6

The preparation from Example 5 was made by winnowing insulin (Novo monocomponent) into cholesterol-sodium lauryl sulfate particles as described in Example 1. The insulin-applied solid emulsifying particles were then coated with a coating solution containing the polyethylene glycol fatty acid ester and egg yolk lecithin with 5% phosphatidic acid (in an amount sufficient so that the final product of 350 mg capsule contained 0.309 x 10_ 2 g). Egg yolk lecithin containing 5% phosphatidic acid dissolved in ethanol was used (Sigmachemical, St. Louis, Mo.).

Test 6

Controlled investigations in a group of IDDM and NIDDM patients were carried out by a Korean research group and a research group from the United States of Korea. The patients received a single oral dose of the preparation of Example 6. The Korean Fosker Group's blood samples were analyzed in Korea and at the same laboratory in the United States as the tests from the United States Research Group's tests. Some of the patients also received ten (10) I.U. regular injectable insulin (Tong-Shin) every other

day. All patients fasted during the examination and the night before. All forms of diabetic therapy were withheld from the patients. Serum samples were analyzed for serum glucose levels. The results (see figures 12 to 21) show that the preparation from example 6 is an orally active insulin for both IDDM and NIDDM patients. Moreover, the results show that both laboratories had similar results.

Regarding figures 12 to 21, the solid lines are investigations carried out by the Korean research group and the dashed lines are the investigations carried out by the research group from the United States. The X-axis is time (h) and the Y-axis is blood sugar mg/dl. Figure 9 shows the results for a 46-year-old woman who received 6 capsules of Example 6. In Figure 10, the subject (38-year-old NIDDM male) received 10 units of regular injected insulin marked x, and then 6 capsules (24 units) from Example 6 .

In Figure 11, the subject, a 47-year-old IDDM male, received 10 units of injected insulin, marked with a ([]), and on day two, 6 capsules (24 units) from Example 6 (x). In Figure 11, symptoms ended, but of less intensity. 11:30, both subjects received 250 ml of pure orange juice and the first subject additionally consumed 250 ml of non-alcoholic drink.

One week later, both voluntarily received the standard breakfast (ie, the breakfast given on the first day of the test) at the same time. 10:00 both subjects received a subcutaneous injection of 16 I.U. Tong-Shin Regular Injectable Insulin-40. About. At 10:30 both subjects began to experience hypoglycemic effects that lasted approximately 2 hours. Both objects were served pure orange juice at 11:20 and were served a standard lunch at 12:00. Blood sugar levels measured at 10:00, 13:00 and 16:00 indicated that the injected insulin produced a hypoglycemic peak effect approx. 3 hours after administration and indicated that, like the oral insulin preparation from Example 1, the effect disappeared 6 hours after administration. (see figure 9) .

Test 1- F

In this test, the oral insulin preparation of Example 1 was administered to 5 subjects suffering from insulin-dependent diabetes mellitus (IDDM) and 5 subjects suffering from non-insulin-dependent diabetes mellitus (NIDDM). The IDDM patients were cared for in a hospital setting for a minimum of three days prior to testing and their blood sugar levels were controlled by a standard hospital diet and a series of subcutaneous injections of regular Tong-Shin insulin. The NIDDM patients had their blood sugar levels controlled by a hospital diet and by oral administration of oral hypoglycemic agents such as chlorpropamide (Diaminase manufactured by Pfizer Laboratories Division, New York, N.Y.).

On the morning of the first day of the test, a normal breakfast was consumed by each of the patients, but the oral hypoglycemic agents and the subcutaneous injections of insulin were withheld from the patients. Instead, the patients received an oral administration of the appropriate dose of the oral insulin preparation of Example 1 and two capsules of the lipolytic enzyme preparation shown in Test 1-E. Blood samples were taken from the subjects immediately before administration of the oral insulin preparation and at two and four hours after administration. These samples were then measured for blood glucose levels. A standard hospital lunch was served to each patient two hours after administration of the oral insulin preparation.

The results of the test are presented in table 1.

As shown in Table 1, blood glucose levels in all patients dropped significantly between administration of the oral insulin preparation and two hours later. However, the sugar levels in the IDDM patients rose again sharply between two and four hours, and in three out of five patients it was roughly equal to or exceeded the levels before treatment. On the other hand, blood sugar levels in those suffering from NIDDM continued to drop between two and four hours in four of the five patients. In the fifth patient, blood sugar levels increased between two and four hours, but did not reach pre-treatment levels.

Example 2

In this example, a prolonged-acting oral insulin preparation was prepared according to the general procedure of Example 1. Particles consisting of cholesterol, sodium lauryl sulfate, and methyl and propyl parabens were prepared and were coated with particles containing Novo monocomponent porcine insulin bound with hydroxypropyl cellulose (40 g hydroxypropyl cellulose in 600 ml water; 6.07% solution), sodium lauryl sulfate, triethylamine HCL, and citric acid. These particles with insulin bound to their surface were then coated with polyethylene glycol monostearate so that a lipid coating was formed around the particles of approximately 0.3

micron thickness in contrast to the approximately 0.1 micron thick coating on the preparations from example 1. The particles were then coated with a gastric juice-resistant coating consisting of the same materials that were used for the preparation in example 1 with a coating thickness of approx. 0.1 micron.

Test 2- A

In this test, the extended-acting oral insulin preparation of Example 2 was administered to three subjects suffering from IDDM and three subjects suffering from NIDDM (subjects A, D, E, G,

I and J from test 1-F). On the morning of the examination, a regular breakfast was consumed by each of the subjects, but the subjects did not receive oral hypoglycemic agents and the subcutaneous insulin injections that were part of their usual therapy. Instead, they were treated with orally administered doses of the preparation of Example 2 and two capsules of the lipolytic enzyme preparation presented in Test 1-E. Blood samples were taken at the time of administration of the oral insulin preparation and at various times thereafter. A standard hospital lunch was served to the subjects four hours after administration of the insulin preparation.

Analysis of the blood sugar levels in the different samples showed that the preparations from Example 2 give a more prolonged hypoglycemic effect than those from Example 1. The preparations with the thicker lipid and gastric juice-resistant coating produced peak hypoglycemic effects after 4 to 6 hours and had effects that lasted up to seven hours , which was the total duration of the survey (see table 2).

Test 2- B

In this test, comparative investigations were carried out on the oral insulin preparation from example 1 and the oral insulin preparation with prolonged action from example 2. Three IDDM patients and three NIDDM patients were administered either the preparation from example 1 or the preparation from example 2 on alternating days together with two capsules of the lipolytic enzymes presented in test 1-E. Two capsules were taken for each oral insulin dose. Each subject fasted overnight and was not served breakfast and was also denied their usual antidiabetic medications. Blood glucose levels for each subject were tested at the time of administration of the oral preparation and at various times thereafter.

The results of administration of the preparations from example

1 and 2 to the IDDM objects, show that the preparation from example 1 produced a strong hypoglycaemic effect within two hours after administration, but that the blood glucose levels returned to approximately their base levels after 4 hours. However, the preparations of Example 2 showed a more gradual effect and did not produce their peak hypoglycemic effect until after 4 to 6 hours. Even after 7 hours they produced a pronounced hypoglycemic effect.

The results of administration to NIDDM subjects show that the differences between the two preparations are less clear. The hypoglycaemic peak effect for both preparations seems to occur after approx. 4 hours, although the preparation from example 2 may have a more prolonged effect than that from example 1. Both preparations gave a more prolonged period of "Normoglycaemic" state in the NIDDM subjects than for the IDDM subjects.

It is suggested that the shorter duration of the hyperglycemic effect of the preparation from example 1 on IDDM patients may be due to the fact that insulin in the preparation is released at a faster rate than that of the preparation from example 2, but that it is also inactivated at a faster rate in IDDM patients due to the presence of anti-insulin antibodies which are often found in IDDM patients. (See figures 10 and 11).

Example 3

In this example, an oral insulin preparation with alternative prolonged action was prepared in accordance with the basic procedure of Example 1. Particles with an average diameter of approx. 50 micron was prepared from a mixture of cholesterol, sodium lauryl sulfate and methyl and propyl parabens. Novo crystalline monocomponent insulin was then bound to the particles with binding agents consisting of hydroxypropyl cellulose, sodium lauryl sulfate, triethylamine HCL and citric acid. The particles with insulin bound to their surface were then coated twice with a polyethylene glycol monostearate coating each 0.15 micron thick. The particles with the double lipid coating were then coated with an enteric coating according to the composition of Example 1 to a thickness of 0.1 micron.

Test 3-A

In this test, the extended-acting insulin preparations from example three were administered to three subjects suffering from IDDM. The treatment was according to the general procedure followed in clinical test 1 where the subjects fasted overnight and were denied their usual hypoglycaemic therapy. The subjects consumed a normal hospital breakfast and were administered an oral dose of the insulin preparation from Example 3 and two capsules of the lipolytic enzyme preparation as presented in test 1-E. The subjects consumed a standard hospital lunch three hours after administration of the oral insulin preparation of Example 3.

The blood glucose levels of the subjects were measured in venous blood samples collected from the subjects at various times after administration of the oral insulin. Data given in Table 3 indicate a prolonged hypoglycemic effect with peak hypoglycemic effect between 4 and 6 hours, but with hypoglycemic effect lasting as long as 9 hours.

A subject suffering from NIDDM was also treated with preparations from Example 3 and lipolytic enzymes as shown in test I-E.

This object fasted overnight before the test as well as during the duration of the test. The NIDDM subject was treated at 10:00 in the morning and her blood glucose level continued to fall during the total 5 1/2 hours that the test lasted.

Example 4

In this example, oral insulin having a composition similar to that of the oral insulin preparation of Example 1 was prepared. The preparation from example 1 was varied by varying the lipid coating. The lipid coating was varied so that in addition to polyethylene glycol monostearate, phosphotidyl phosphate (in an amount sufficient to make a final concentration of 0.046 x 10-<2>g), phospholipids (in an amount sufficient to make a final concentration of 0.046 x 10-<2>g) and cholesterol (an amount sufficient to make a final concentration of 2.628 x 10-<4>mol) added to the lipid coating materials. This lipid coating was coated on the insulin and particles to a thickness of 0.3 microns. 240 mg of the oral insulin product was then packed by hand into a hard gel capsule. The gel capsule was coated with a gastric juice-resistant coating from example 1.

Example 5

In this example, oral insulin corresponding to the oral insulin preparation from example 3 was prepared. The oral insulin is modified to the extent that a combination of cholesterol-arginine HCL was used instead of cholesterol in Example 3. Cholesterol-arginine HCL provides a mixture that mimics the low specific gravity lipoprotein mixtures in vivo. Also, the lipid coating materials were changed. After coating the mixture with a lipid coating corresponding to that in Example 4 in a thickness of 0.15 microns, a second lipid coating is applied. The second lipid coating comprises oleic acid (an amount sufficient to give a final concentration of 8.776 x IO-<6>mol) and tromethane (an amount sufficient to give a final concentration of 8.764 x IO-<6>

mol) as "vulcanizing agent" or "softening agent" of the liquid coating membranes.

Briefly, the oral insulin in this example consists of "low specific gravity lipoprotein-like" solid emulsifiers as a base. NOVO Monocomponent crystalline porcine insulin was bound to the emulsifier and the resulting product was coated with a 0.1 micron thick hydroxypropylmethylcellulose water-soluble coating, which was then coated twice with the lipid coating materials to a final membrane thickness of 0.3 micron (0.15 micron per layer of the lipid coating), a "stabilizing tromethane" was added, and the product was then packed by hand into hard gel capsules (so that each capsule contained 240 mg of the mixture). The capsule was then coated with the enteric-resistant material from Example 1. The water-soluble coating allows the insulin-cholesterol arginine HCL to be coated with a lipid coating.

Test 5- A

The bioavailability of insulin and its corresponding hypoglycemic effect in the oral insulin preparations from example 4 and example 5 were investigated on a group of IDDM and NIDDM. Oral insulin from Example 4 was also examined in two normal, healthy male volunteers. In all tests presented in 5-A, the subjects received with each dose of oral insulin two capsules of the lipolytic enzyme from test 1-E.

At least 12 or 24 hours before the examination, the patients' usual anti-diabetic treatment agents were withheld from the patients. After overnight fasting and fasting throughout the study period, oral insulin preparations from Example 4 or Example 5 were orally administered.

A venous sampling catheter was placed in a vein, usually in the anti-cubital vein and blood samples were taken at time points 0, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0 and/or 5.0 hours after drug administration. Serum glucose was measured on a Gilfordx400, and serum insulin was analyzed using an "immuno-enzymatic o-phenylenediamine (OPD)" method and insulin-B test kit from WAKO (Japan). All measurements were made in duplicate.

As indicated above, two healthy male volunteers, aged 38 and 31 years, participated in the study. A subject received the oral insulin of Example 4 twice over a two-week period, once each week, receiving 0.20 and 0.27 I.U. per kilos of the oral insulin. The subject's serum glucose level decreased from a baseline of approx. 111.0 mg/dl to an average of 80.5 mg/dl 3 hours after the time of drug administration. The serum insulin level rose to 20 micro-U per ml, which occurred 1 1/2 hours after taking the drug in both cases. 45 minutes after administration of the oral insulin, the subject complained of blurred vision, cold sweats, irritability and poor ability to concentrate.

The second volunteer received 10 I.U. regular injectable insulin-40 (from Tong-Shin Pharm. Co. of Seoul, Korea), subcutaneously. Typical insulin-induced shock symptoms were observed 72 minutes after injection of regular insulin, serum glucose was 47 mg/dl at that time. He was given a glass of fresh orange juice and the serum glucose level rose to va. 115 mg/dl within 1 hour. 5 IDDM patients have completed all three phases of the examination in test 5-A, i.e. they received 10 I.U. subcutaneous injection of regular injectable insulin-40 (from Tong-Shin Pharm. Co. of Seoul, Korea) in the first week; they received a single oral dose of Example 4 (0.544 I.U./kg mean dose) one week later; and the following week they received 9 consecutive doses of Example 4 (average dose was 0.544 I.U./kg) given 6 hours apart.

The oral Insulin from Example 4 was administered to a total of 8 IDDM patients (10 trials) (2 repeated the study) and 7 NIDDM patients (11 trials) (4 patients repeated the study). 7 IDDM patients completed the study after oral administration of a single dose of the oral insulin preparation of Example 5.

The investigations showed that oral insulin from example 4 apparently has a later onset of action, but shows a longer duration of "hypoglycemic" action for both IDDM and NIDDM patients, compared to regular insulin. The bioavailability for insulin from the preparation in example 4 was approx. 3 times that of regular insulin; however, when adjusting for insulin dosage, it was about the same (See Table 4).

The investigations also showed that the insulin preparation from

example 5 produced an onset of hypoglycemia and had an overall duration of hypoglycemia that was broadly similar to that observed after subcutaneous injections of regular insulin. The bioavailability of insulin was greater when Example 5 was taken orally than Example 4.

Co = Basal value for serum glucose; Cm in = Minimum value for glucose; Tc -min = time for observation of the minimum glucose level; Tioo = time to reach 100 mg/dl g; serum glucose; dC/dT = rate of decrease in serum glucose levels. Io = basal value of serum insulin; Ima x = maximum value for insulin; Ti-max = time to reach the maximum serum insulin level; Ti -o = the time to reach the baseline value of serum insulin level; dl/dT = rate of rise of serum insulin level; AUC = the area under the serum insulin level curve over the (examination) time. (<*>two peaks in serum insulin levels).

Example 6

The preparation from Example 5 was made by winnowing insulin (Novo monocomponent) into cholesterol-sodium lauryl sulfate particles as described in Example 1. The insulin-applied solid emulsifying particles were then coated with a coating solution containing the polyethylene glycol fatty acid ester and egg yolk lecithin with 5% phosphatidic acid (in an amount sufficient so that the final product of 350 mg capsule contained 0.309 x IO-<2>g). Egg yolk lecithin containing 5% phosphatidic acid dissolved in ethanol was used (Sigmachemical, St. Louis, Mo.).

Test 6

Controlled investigations in a group of IDDM and NIDDM patients were carried out by a Korean research group and a research group from the United States of Korea. The patients received a single oral dose of the preparation of Example 6. The Korean Fosker Group's blood samples were analyzed in Korea and at the same laboratory in the United States as the tests from the United States Research Group's tests. Some of the patients also received ten (10) I.U. regular injectable insulin (Tong-Shin) every other day. All patients fasted during the examination and the night before. All forms of diabetic therapy were withheld from the patients. Serum samples were analyzed for serum glucose levels. The results (see figures 12 to 21) show that the preparation from example 6 is an orally active insulin for both IDDM and NIDDM patients. Moreover, the results show that both laboratories had similar results.

Regarding figures 12 to 21, the solid lines are investigations carried out by the Korean research group and the dashed lines are the investigations carried out by the research group from the United States. The X-axis is time (h) and the Y-axis is blood sugar mg/dl. Figure 9 shows the results for a 46-year-old woman who received 6 capsules of Example 6. In Figure 10, the subject (38-year-old NIDDM male) received 10 units of regular injected insulin marked x, and then 6 capsules (24 units) from Example 6 .

In Figure 11, the subject, a 47-year-old IDDM male, received 10 units of injected insulin, marked with a ( []), and on day two, 6 capsules (24 units) from Example 6 (x). In Figure 11, the subject, an IDDM 45-year-old male, received ten units of injectable insulin ([]) and on day two 8 capsules (32 units) from Example 6 (x).

In Figure 12, a 47-year-old NIDDM woman received 6 capsules, 24 units from Example 6. In Figure 13, an IDDM patient received 14 capsules, 56 units from Example 6. In Figure 14, an IDDM patient received 12 capsules, 48 units from Example 6. In Figure 15, a 38-year-old IDDM male patient received 10 units of injectable insulin ([]), and on day two, 10 capsules, 40 units from Example 6 (x). In Figure 16, a 47-year-old man with IDDM received 10 units of regular injected insulin ([]) and on day two 8 capsules (32 units) from Example 6 (x). In Figure 17, a 49-year-old male IDDM patient received ten units of regular injected insulin ([]) and on day two, 6 capsules (24 units) from Example

6 (x) .

All subjects received two capsules of the lipolytic enzymes from Test 1-E with each dose of the preparation from Example 6.

Example 7

To prepare the preparation of Example 7, cholesterol (Fuluka, A.G.; Switzerland) and L-arginine HCL (WAko Jun-Yaku, Japan; KWJ 3778) were thoroughly mixed in a ratio of two to one in a vessel. Gum arabic (Yakuri-Kagaku, Japan; A001 = 03) was used as a filler.

A lipid (Cholesterol) and an amino acid (L-arginine HCL) were linked to hydroxypropyl cellulose containing insulin (Novo monocomponent Porchin Crystalline insulin). Citric acid was used as an inhibitor of the insulin-deactivating enzyme and for pH adjustment. Triethylamine HCL was used as an insulin stabilizer and as an anti-foam agent.

The procedure for combining the insulin with the particles was the same as Example 1 with the spray air flow rate set to 12 L/min and the spray air pressure to 2.5 kg/M<2>.

The lipid and amino acid particles combined with the crystalline insulin were coated with a water-soluble membrane consisting of hydroxypropylmethylcellulose (Shin-Etsu-Kagaku, Japan). The above lipid-amino acid particles combined with insulin and coated with a water-soluble membrane were coated with a lipid coating consisting of the polyethylene glycol fatty acid ester, egg yolk lecithin containing 5% phosphatidic acid and oleic acid (Sigma Chem., St. Louis). Tromethamine was used as a vulcanizing agent.

The resulting preparation was filled into a hard gel capsule and coated with a gastric juice-resistant coating as in example 1.

Each of the resulting 240 milligram capsules contained:

The preparation from example 7 produces an effect equivalent to a subcutaneous injection of regular insulin in humans.

It is believed that the following limits can be used for a preparation made in accordance with Example 7. Each resulting 240 mg capsule can consist of:

Novo monocomponent Porchin Crystalline insulin or other insulin is added so that each capsule contains between 4 and 40 international units/capsule of insulin.

Methylparaben (3.179 to 7.803) x 10"<5>M, propylparaben (0.809 to 1.681) x 10-<5>M and silica gel (0.275 to 0.641) x 10_<2>6 may be added to the final powdered coated oral insulin particles.

The final, coated oral insulin powder can be filled in a hard gel capsule, soft gel capsule or in a hard pressed one

tablet form. These forms can be given an enteric-resistant coating.

Example 8

A preparation was made following the recipe from Example 7, however, the resulting recipe was modified as follows:

Each of the above capsules containing 240 mg can consist of:

Test 8

Seven insulin-dependent diabetic patients (IDDM) 6 men and one woman between 24 and 47 years old (mean 34.4) weighing between 48 and 68 kg each (mean 58.6 kg) took between 32 and 48 (mean 42.6) international units oral insulin preparation from example 7, orally after fasting overnight and fasting during the examination.

5 male IDDM patients received 0.172 IU/kg regular injectable insulin (Tong-Shin) subcutaneously after fasting overnight and during the examination in new 5 male IDDM patients, the total effect of the oral insulin preparation from example 7 in terms of reducing blood sugar levels is similar to that of regular injectable insulin (see Figure 22).

CO = baseline value for glucose;

<c>min = minimum glucose value observed;

<T>c-min = The time of observation of the minimum

glucose level;

T100 = time to reach 100 mg/dl;

dC/dT = rate of hypoglycaemic action.

In two IDDM patients, the bioavailability of insulin (insulin levels in serum) was measured after oral administration of the oral insulin preparation from Example 7 (0.63 IU/kg).

Compared with 0.172 IU/kg of regular insulin, subcutaneous injection to 5 IDDM patients, the "pharmacokinetics and bioavailability" of insulin are as follows:

Insulin-

preparation 0914

Io = basal insulin value;

Ima x = maximum value for serum insulin;

Ti-max = time for observation of maximum value of serum insulin;

Ti -o = Time to reach baseline value of serum insulin;

dl/dT = rate of rise of serum insulin (absorption rate);

AUC = the area under the curve for serum insulin level over the examination period.

Each subject in test 8 received two capsules of the lipolytic enzyme preparation presented in test i-E with each dose of oral insulin.

The clinical investigation showed that the oral insulin preparation from Example 7 at a single oral dose of 0.630 IU/kg insulin produced an identical "hypoglycemic" (or effect by reducing serum glucose) effect in diabetics as that after subcutaneous injection of 0.172 IU/kg insulin normally injectable insulin (Tong-Shin) to another group of insulin-dependent diabetics. 4 of our 7 investigated insulin-dependent diabetics experienced an insulin shock corrected by oral intake of sugar water and fresh orange juice.

However, pharmacokinetic and bioavailability data after 0.630 IU/kg oral insulin preparation of Example 7 compared with 0.172 IU/kg subcutaneous injection regular injectable insulin (Tong Shin) indicated that the absorption rate of insulin was more than twice as fast after oral insulin as after

injection of regular insulin. The bioavailability of insulin (AUC) after oral insulin (0.630 IU/kg) is almost 4.7 times the AUC after subcutaneous injection of regular insulin (0.172 IU/kg). AUC after adjusting the administered insulin dose according to

oral insulin is 2,264.8/IU and it is 1,175.0/IU after subcutaneous injection of regular insulin. The bioavailability of the oral insulin preparation from example 7 is equal to or greater than that of ordinary injectable insulin, per given insulin dose.

The effect of the oral insulin preparation from example 7 in terms of reducing serum glucose levels in diabetics and the bioavailability of insulin for diabetics after oral administration of the oral insulin preparation from example 7 is excellent and is equivalent to that of subcutaneously injected ordinary injectable insulin in insulin-dependent diabetics.

Example 9

In this example, the lipolytic enzymes are lipid and enteric coated and can be combined with an oral insulin preparation in a single capsule or tablet.

A. Pancreatic lipase.

Pancreatic lipase particles were prepared as follows. The following components were filled in the modified Spir-A-Flow previously described.

The components were placed on the disc in Spir-A-Flow1 and dried at a temperature that varied between approx. 30 and 32°C while the rotor was rotated at 300 rotations per minute. minute (rpm), the stirrer at 500 rpm, and the chopper at 2,000 rpm. Fine dried particles of the components were thereby formed and were blown from the bottom of the lower part by air heated to a temperature of approx. 30 to 32°C. The heated air was blown through the gap created between the rotor and the wall of the vessel (gap air) and through the fine mesh openings of the rotating disk (air flow). Both the slit air and the airflow were at an air pressure that varied between 100 and 450 mmH2 0. The fine particles of the components were suspended inside the vessel of the Spir-A-Flow by the slit air and airflow. The particles of the components were then coated with the following coating solution:

To coat the particles with the coating solution,

the coating solution was sprayed onto the particles of the components after they were suspended inside the vessel, at a rate of 10 ml/min and with a pressure of between 1.0 and to 5.0 kg/M<2> for 120 minutes (the spray nozzle was alternating "on" for 20 to 40 seconds and "off" for 60 to 90 seconds).

This resulted in the pancreatic lipase particles being coated and thereby shielded by a thin layer of water-soluble HPMC-L membrane. These particles of pancreatic lipase coated with an HPMC-L membrane were then enteric coated using the following enteric coating solution:

The gastric juice-resistant coating solution had a pH of approx. 5.2 to approx. 5.6.

Alternatively, a commercial grade acrylic resin called EUDRAGIT<R>L-100-55 (or L-100) or EUDRAGIT"-L30-D-55 can be used to coat the particles.

The particles were enteric coated using the first mentioned enteric coating solution by spraying the enteric coating solution on the HPMC-L coated particles of pancreatic lipase, while the particles were again suspended in the vessel, at a spray rate of 10 ml/min and a pressure of 2 .5 kg/m<2>in 3 (three) to 4 (four) hours. The spray nozzle was "on" for 20 to 40 seconds and "off" for 60 to 120 seconds. The resulting product provides the desired dissolution time in "gastric dissolution" and in "duodenal dissolution".

B. Bile salt.

Bile salt particles were formed by a Spir-A-Flow and coated bile salts and avicel with the coating solution and the aforementioned gastric juice resistant coating solution used to coat the pancreatic lipase above. The bile salt and avicel were coated as shown in A above. Bile salt and the avicel components used were the following:

C. Oral insulin preparation

The oral insulin preparation was made in the following way. The following components were mixed in Spir-A-Flow.

The components were carefully mixed and dried in the tub of the Spir-A-Flow. The following parameters were used in the Spir-A-Flow: The rotor was set at 350 rpm; the stirrer was set to 500; the chopper was set at 2,000 rpm; the gap air pressure was between 100 and 300 mmH2 0; the airflow pressure was between 100 and 350 mmH2 0; the air was at a product temperature of 24 to approx. 31°C; The components were mixed and dried in Spir-A-Flow1 with these parameters for approx. 30 minutes. Then, carefully mixed and dried components suspended in the air in the vessel were combined with the following binder solution.

To coat the particles formed above, the binder solution was sprayed through the spray nozzles located at the top of the Spi-A-Flow1 one, see 28 in Figure 3, the binder solution was placed in a cylinder and then cooled by immersing in an ice-cold beaker before and during the spraying process. The binding solution was applied to the fine air-suspended particles at a rate of 10 to 14 ml/min, at an air pressure of 1.0 to 2.0 kg/m<2>and an air volume of 15 to 25 L/min, for approximately 2 hours; until all the binding solution has been used. The spray nozzle was alternately turned "on" for 20 to 40 seconds and "off" for 30 to 90 seconds. During the process, the product temperature was kept between approx. 25 and approx. 32°C by keeping the air used to suspend the particles at this temperature. The resulting particles with the crystalline insulin were then instantly dissolved in water, which has a temperature of approx. 24°C, to create an emulsion with insulin bound to the fine particles. The emulsion has a pH of approx. 4.2 to about 5.4.

Immediately thereafter, the insulin-bound particles in the emulsion were coated with a lipid coating solution prepared as follows. The following components make up the lipid coating that was sprayed onto the particles with insulin on them.

The lipid coating solution was also sprayed through spray nozzle 28 located on top of the vessel. The parameters of the Spir-A-Flow1 one were as follows: the rotor was set at 250 to 300 rpm; the stirrer of 500; and the chopper at 2,500 to 3,000 rpm. The spray nozzle and its spray rate were set at 12.0 ml/min at an air pressure of 1.2 to 1.5 kg/m2 , an air volume of 15 to 20 L/min and the product temperature was between approx. 30 and 34°C. Again, the product temperature is maintained by setting the temperature regulator on the Spir-A-Flow1 so that the air flowing into the tub is at the desired temperature. The lipid overcoating solution was sprayed for 60 minutes. The spray nozzle is alternately turned "on" for 20 seconds and "off" for between 60 and 90 seconds.

The resulting lipid coated particles are then recoated with a lipid coating solution for recoating. The lipid recoating solution for recoating contains the following components.

The lipid solution for recoating was sprayed from the side of the Spir-A-Flow tub at a spray rate of 12 ml/min at an air pressure of 1.0 to 1.5 /m<2> and with an air volume of 15 to 20 L/min. The rotor was set at 300 to 350 rpm, the stirrer at 500, the chopper at 3,000 to 3,500 rpm and the temperature of the product was maintained at approx. 31 to approx. 35°C. The spray nozzle was alternately turned "on" for 20 to 40 seconds and shot "off"

for 60 to 90 seconds.

If desired, trometamol can be omitted from the lipid solution for renewed coating. However, if trometamol is omitted, drying time will be increased as a result and extreme care will have to be taken to prevent "too greasy, moist" particles. Tromethamine (trometamol) is listed in the U.S. and European Pharmacopoeia.

After the repeated lipid coating solution step, the coated dried particles are either filled into hard gel capsules or pressed into tablet form. The oral insulin-linked coated particles can be made into capsules or tablets alone or can be mixed with the enteric particles of lipase and bile salts previously described. The resulting capsules or tablets can be enteric coated.

Alternatively, the lipid-coated binding oral insulin particles themselves can be given a gastro-resistant coating. The gastric juice-resistant coating solution for coating the lipid-coated oral insulin particles comprises the following components:

To form enteric coated lipid coated oral insulin particles, the enteric coating solution was sprayed through spray nozzle 28 located at the top of the vessel on the Spir-A-Flow at a spray rate of one gram kilogram per minute. minute to about 500 grams of the lipid-coated binding oral insulin particles. The particles were sprayed for 5 seconds and allowed to dry for five seconds, alternating until the desired thickness of the enteric coating was achieved.

To determine the amount of enteric coating solution to be applied in percent dry coating substance (enteric coating solution) ("Q"), the surface area of the particle to be coated (SA) in mm<2> was multiplied by mg of dry varnish (L) per . Cm<2>desired in mg/Cm<2>(thickness of the coating), this is divided by the weight of oral insulin-binding particles (W) in mg. In accordance with this, the percent dry lacquer substance "Cj" is calculated by the following equation:

Preferably, the enteric coated particles of the oral insulin have a diameter between approx. 0.1 and approx. 0.8 mm (the surface area thus varies between approx. 0.314 and approx. 2.0 mm<2>). It has been found that by spraying approx. 120 to approx. 150 ml of the above enteric coating solution on oral insulin particles weighing approx. 450 grams, the resulting coated particles will not dissolve when placed in a solution having a pH of about 1.2 (a gastric solution) for at least 10 minutes; in the meantime, the coated particles will dissolve and disperse in a solution having a pH of approx. 6.8 (a duodenal solution) within 30 seconds. In accordance with this, an ideal gastric juice-resistant coating has been achieved for the oral insulin particles.

D. Oral insulin particle/ lipase/ bile salt capsule

In this example, capsules have been made which comprise the oral insulin-coated particles, coated lipase and coated bile salt. To this end, to each #l size hard-gel capsule, size one, are mixed the enteric-coated oral insulin particles (prepared as described above in C), enteric-coated lipase particles (prepared as described above in A) and enteric-coated bile salt particles prepared as described above in B) together and filled into hardgel capsules in the following proportions:

200 Mg (12 IUS) oral insulin

28 Mg (1,000) units of pancreatic lipase

25 Mg of bile salt

50 Mg sodium bicarbonate (oven dried);

and

50 Mg citric acid as "filler".

Sodium bicarbonate and citric acid produce a bursting effect that causes the hard gel capsule to open in the stomach, releasing the enteric-coated particles of oral insulin, lipase and bile salts in sizes less than 1 mm. Because of their size and composition, these particles quickly pass through the pylorus into the duodenum. The sodium bicarbonate and citric acid also act to adjust the pH in the intestine towards alkalinity and increase the activity of the pancreatic lipase in the presence of the bile salt.

E. Comparative clinical examination of the oral insulin preparation from C compared to the oral insulin preparation from D.

Six diabetic subjects (three insulin-dependent and three non-insulin-dependent; 5 men and 1 woman) participated in a comparative study between the oral insulin preparation from C given together with 4 capsules of the lipolytic enzyme, each containing 1,000 units of lipase and 25 mg of bile salt, and the oral insulin preparation from D, i.e. the oral insulin preparation from C together with the coated lipase from A, coated bile salts from B, sodium bicarbonate and citric acid in a hard gel capsule.

Each patient received each preparation on different days. For this purpose, each patient orally received 48 IU of oral insulin with 4 capsules containing 4,000 units of lipase and 100 mg of bile salt on the first day of the study and 4 capsules containing 48 IU of oral insulin and 4,000 units of lipase and 100 mg of bile salt on the second day of the study. On the third and fourth day of the examination, the routine was repeated.

In general, whether the lipolytic enzyme capsules were given separately but at the same time as the oral insulin (48 IU) capsules or the lipolytic enzymes were contained in the same capsule as the oral insulin and given as a combined capsule, the overall effect on blood glucose and hypoglycemic course/effect was of the oral insulin the same.

F. Oral insulin/ lipase/ bile tablet:

The non-enteric coated particles described in Sections A, B and C can be collected and made into a hard pressed tablet. For this purpose, the following components were mixed. Insulin/lipase/bile salt particle (in the same

The above mixture was prepared in hard pressed tablets of approximate size 220 mg (if desired the size can be more or less). If crystalline insulin is substituted in the preparation from section C, each tablet may contain 6 IU of insulin; The insulin dose can be increased up to 40 IU if desired.

The resulting tablets were coated with EUDRAGIT"-E-100

in a final "film coating" thickness of approx. 0.1 to approx. 0.15

etc. Tablets coated in this manner were found to dissolve in gastric juice solution having a pH up to 5.0 within 30 seconds or less. The particles thereby released, i.e. oral insulin, lipase and bile salts, because they have become enteric coated, dissolve only easily in a solution having a pH of 6.0 or more. Moreover, the particles will have a size of less than 1 mm and will therefore easily pass through the pylorus and dissolve in the duodenum. In the duodenum, the particles are emulsified and suspend the microlipospheric particles that have insulin bound to them (having a size of less than 1 micron) and/or "lipid layer coating" - micro-lipospheric-bound insulin (having a size of less than 50 microns). In accordance with this, the particles will be suspended in an emulsion that is formed inside the cavity of the duodenum. These particles are thereby preferentially absorbed from the tufts on the intestinal mucosa and pass by the liver.

Clinical investigations of the tablet form of the oral insulin/lipase/bile salts were carried out on three insulin-dependent diabetics and the results were as follows:

The use of a stomach-resistant coating solution such as EUDRAGIT"-L 100 or L-30 to coat the oral insulin particles, bile salt particles and lipase particles produces coated particles with the following characteristics; 1. Resistance to gastric juice; 2. Protects the inside of the stomach against possible etching caused of the particles 3. Isolates incompatible drugs 4. Improves shelf life of products;

5. isolates hydroscopic cores and

6. protects the particles from atmospheric influences.

As presented above, EUDRAGIT" protects the wall of the stomach from any corrosive effect that the bile salts may have and also improves the oral insulin preparations which have poor shelf life and are extremely labile and hydroscopic.

Example 10

According to this example, an oral insulin preparation was prepared. The following components were placed in the tub of the Spir-A-Flow.

These powdered components were ground using the Jet Mill previously described (see Figures 1 and 2) to a particle size of less than 50 microns, and carefully mixed and dried in the vessel of the Spri-A-Flow using the following parameters:

The carefully mixed and dried components (particles) were then suspended in the vessel by increasing the gap air pressure and the air flow pressure to approx. 50 to approx. 100 mmH2 0. The suspended, fine, dried particles were then coated with the following solution: The binding solution is sprayed from the spray nozzles located on top of the vessel so that it covers the particles. The binding solution containing the insulin crystals is bound to the fine, dried particles by using the following parameters;

The particles were sprayed alternately by an approx. 90 to a approx. 120 minute period, until the entire total solution was used. The particles were sprayed for approx. 5 seconds and dried for approx. 22 seconds alternating during the period.

The particles coated with crystalline insulin solution (binding particles) were then coated with a first coating solution. The first cover solution included the following:

The above coated particles were then suspended in the Spir-A-Flow and were coated with the first coating solution above under the following conditions:

Bound particles were sprayed for approx. 5 to approx. 10 seconds, and dried for approx. 20 to approx. 22 seconds.

The coated bonded particles were then recoated with a different coating solution. The second coating solution contained the following:

The second coating solution was sprayed onto the suspended coated bound particles from the spray nozzle located on the side of the vessel using the following parameters:

The resulting coated bound particles, comprising insulin and coated with lipids, can either be prepared as fine granules having a size of less than 2 mm in diameter (using NaHCO 3 : citric acid in a 3:1 ratio as a "seed" coated with starch, which is coated with a coating layer, and insulin-coated chemical particles described above as the outer layer, etc., and then coated with an enteric coating), or mixed with sodium bicarbonate, 20 to 30 percent by weight; citric acid 10-20 percent by weight and silica gel 3 to 6 grams per 500 - 650 grams final weight of the preparation, and is filled in size no. 4 hard gel capsules. The resulting capsules were then enteric-resistant coated with the following enteric-resistant coating solution: Shellac 0.2 g per 100 ml of the above mixed solution is added and dissolved (dark brown color) pH = 5.2 to 5.6; mixed at a temperature of approx. 30.0 to approx. 32.5°C Alternatively, an enteric coating solution containing the following components can be used;

The first mentioned enteric coating solution was used to enteric coat the size #4 capsule filled with the coated insulin particles. The coated insulin particles in the size #4 capsule so coated were found to dissolve only in a gastric juice solution having a pH of about 1.2 after exposure for two or more hours. However, the capsules dissolve in duodenal fluid which has a pH of approx. 6.8 within 15 (fifteen) minutes or less, (as specified in the U.S. Pharmacopea). If the capsules coated with the alternative enteric coating solution were also found to have similar properties.

Example 11

To prepare the oral insulin preparation in this example, the following components were filled into the vessel of the Spir-A-Flow;

After the components were mixed and dried in the vessel of the Spir-A-Flow, using the parameters described in Example 10 (above), these particles were bound with an insulin-containing binding solution. The insulin-containing binder solution contained the following:

The mixed and dried components were suspended in the vessel and the insulin-containing binding solution was sprayed from spray nozzle 28 located at the top of the vessel. The binder solution was sprayed while the Spir-A-Flow was operating at conditions set forth in Example 10 above.

The insulin-bound particles are then coated with a first coating solution. The first coating solution contained the following chemicals;

The coated insulin bound particles were coated with a first coating solution as described in Example 10 (above).

The resulting coated particles were then recoated with a different coating solution. The second coating solution had the following components;

The second coating solution was sprayed from the spray nozzles 27 placed on the side of the vessel under the conditions described in example 10 (above).

The resulting coated particles can be prepared as fine grains less than 2 mm in diameter containing sodium bicarbonate and citric acid, in a 3:1 or 1:1 ratio mixture, as germ; starch such as "nonpareil"; coated with a coating layer; insulin-binding particles as the outer layer; and gastric juice-resistant coating or mixed with approx.

20 to approx. 30 weight percent sodium bicarbonate, approx. 10 to approx. 20 percent by weight citric acid and approx. 0.5 to approx. 2.0% by weight of the silica. The resulting mixtures can then be filled into No. 4 hardgel capsules, and coated with an enteric coating.

Example 12

In this example, the oral insulin preparations made in examples 10 and 11, which consist of grains with a diameter of less than 2.0 mm, are coated. A CF granulator (See figure 23) is used and the oral insulin preparation from either 10 or 11 was mixed with sodium bicarbonate/citric acid in a ratio of approx. 3:1 that has a size that has a size of less than 0.5 mm is used as "nonpareil". The resulting grain was then enteric coated with one of the previously described enteric coating solutions. It was found that the resulting granules would not dissolve for at least a two hour period in gastric juice solution having a pH of about 1.2, but will dissolve within 15 minutes or less in duodenal solution which has a pH of approx. 6.8.

The CF granulator commercially available through Freund Industries, Inc., Tokyo, Japan, is designed to collect, granulate and coat using a centrifugal system and is shown in Figure 23. Briefly, the CF granulator comprises a power supply 42, rotor 44, air supply 46 and spray nozzle 48.

To form products in the CF granulator, the temperature of the air supply was set between approx. 28 and approx. 34°C, the rotor speed was set between approx. 130 and 200 rotations per minute, the air supply flow rate was 30 to 50 L/minute, the spray nozzle's spray rate was 20 to 50 ml/min at a spray pressure of 10 to 30 L/min.

To form the sodium bicarbonate/citric acid mixture, citric acid, 110 to ca. 150 g, as "seed" sodium bicarbonate powder, approx. 400 to approx. 450 g, and a small amount of "alph" starch powder, approx. 10 to approx. 50 g, covered with citric acid and dried for thirty minutes in Spir-A-Flow.

Using citric acid as seed, and sodium bicarbonate and starch as coating (both as "nonpariel"), oral insulin preparations of Examples 10 and 11 as the powder are premixed with approx. 30 weight percent starch, coated on a granular "nonpareil" using about 6 to about 20% (preferably about 10%) hydroxypropyl cellulose-L (HPC-L) in alcohol at a rotor speed of about 100 to approx. 200 rpm, an air supply of 20 to 50

L/min, a spray nozzle flow rate of 15 to 40 ml/min at

a pressure of 10 to 30 L/min for 20 to 40 minutes (the temperature was about 25 to about 30°). The resulting grains were dried in a Spir-A-Flow for 30 minutes at a temperature of approx. 30 to approx.

35° C.

The resulting granular preparation had grains with a diameter of between approx. 0.490 and approx. 1,450 mm and was gastric juice resistant coated using a gastric juice resistant solution of 5% HPC/methyl phthalate and polyethylene glycol-6000 (0.5%) dissolved in ethanol. Other enteric coating solutions indicated above could also have been used. The grains were coated with the enteric solution at a rotor speed of approx. 150 to 250 rpm, air supply of 30 to 50 l/min, spray flow of 20 ml/min at pressure of 15 to 30 L/min, and at a temperature of approx. 25 to approx. 30°C. The resulting grains have a structure roughly similar to that shown in Figure 24. The grain 50 comprises a core or seed 52, a "nonpareil" 54, an HPC-L coating 56, a layer of insulin 58, and an enteric coating 59. These grains will does not dissolve for at least two hours in gastric juice solution with a pH of approx. 1.2, but will dissolve within 15 minutes or less in duodenal solution with a pH of approx. 6.8.

Test 12 a

The resulting product, approx. 0.9 (0.69 to 1.07) IU per kg, was dissolved in 10 to 20 ml of saline, in vitro, and administered to the duodenum of three dogs via a Wallace-Diamond double-cavity tube. Series of blood samples from the femoral vein were collected at time 0, 60, 80, 100, 180, 240, and 300 minutes after administration of the preparation to three dogs. The dogs, one male and two females, had an average weight of 4.0 (approx.

3.0 to approx. 5.6) kilos each. Plasma glucose levels and insulin levels

(by radioimmunoassay and Wako's enzyme immunoassay methods)

was measured.

A marked and significant degree of hypoglycaemia was observed after approx. 80 minutes, which lasted approx. 300 minutes, while a maximum plasma insulin level was observed at approx. 100 minutes after the intraduodenal administration of the preparation (See figures 25 and 26).

Test 12 b

The preparation from example 11 changed as indicated in example

12, approx. 1.0 IU per kg, was administered intraduodenal to a 6.0 kg female dog. The hypoglycaemic effect of the preparation was compared with the intraduodenal administered regular injectable insulin (Novo aktrapid insulin, 1.0 IU per kg) and intravenously administered regular injectable insulin (Novo aktrapid insulin, 0.5 IU per kg). After infusion of 1.26 IU regular injectable insulin intravenously, plasma glucose level fell from 102 to 23 mg/ml within 20 minutes and returned to 100 mg/ml within three hours. Then, 6 IU of regular injectable insulin was given intraduodenal. No change in blood glucose levels was observed for more than a three-hour period. Six (6) IU of the preparation was given intraduodenal, and a significant and persistent reduction in blood sugar level was observed. The animal died 2 hours after administration of the preparation due to severe hypoglycaemia (See Figure 27).

Test 12 c

After a 24-hour observation period, without insulin or oral hypoglycaemic agents, and fasting overnight, three insulin-dependent and two non-insulin-dependent patients took approx. 0.64 to approx. 0.73 IU per kilograms of the insulin preparation from Example 10 changed as indicated in Example 12. Blood sugar levels were measured every hour for 5 hours. To one IDDM and one NIDDM patient, however, due to the lower fasting blood sugar levels observed before the examination, a standard breakfast was given and the examination was continued, after the meal. A clinically significant degree of "hypoglycemia" was observed in these patients (see Table 5 below).

6 new patients, 4 IDDM and 2 NIDDM patients, took 0.49 to 0.63 IU per kilo of the preparation from example 11 changed as indicated in example 12, orally. One patient, because lower fasting blood sugar levels were observed, received a standard breakfast and the examination was continued. In the remaining 5 cases, the examination was carried out under fasting conditions. A significant degree of hypoglycaemic effect was observed after the oral administration of the preparation (see table below).

12 d clinical investigations with granular preparations i

diabetic examination.

A group of diabetics participated in investigations with testing

of the preparations from examples 10 and 11 twice during a two-week period. During the first week, each patient orally took either oral insulin from Example 10 or Example 11 in size #4 enteric-coated capsules, and one week later the same patient took an alternative granular preparation of either Example 10 or Example 11 orally . The granular preparation from Example 11 orally administered an average insulin dose of 0.145 units per kilogram of body weight (0.11 to 0.18 units per kilogram of body weight) and produced a more rapid reduction in blood glucose levels over a measured two-hour period compared to the oral administration of the preparation of Example 11 in size #4 capsules which had an average insulin dose of 0.583 units per kilogram of body weight. kilogram of body weight (0.49 to 0.63 units per kilogram of weight); which is almost 4 (four) times the dose of the example 11 preparation in granular form. Meanwhile, the formulation of Example 11 in size #4 capsules lasted longer and continued to exhibit its hypoglycemic action and efficacy beyond the two hour time point after dosing (see Figure 28). The degree of hypoglycemia experienced 1 hour after dosing the preparation from Example 10 was somewhat greater when the oral insulin preparation was given in granular form than in size #4 hardgel capsules. However, the oral insulin preparation of Example 10 in size #4 hardgel capsules has a relatively longer lasting hypoglycemic effect than the granular form of the oral insulin preparation of Example 10 (see Figure 28).

Thus, ideally a mixture of the preparation from example 10 in granular form and in size #4 hard gel capsule form, or alternatively a mixture of the preparation from example 11 in granular form and in size #4 hard gel capsule form can provide a rapid and yet longer lasting hypoglycemic effect on diabetics . In fact, by administering the oral insulin preparation of Example 11 in granular form (average 0.164 units per kg) and in size #4 hardgel capsule preparation (average 0.566 units per kg) a more rapid onset and longer lasting hypoglycemic effect was observed in these diabetics (see figure 28).

Alternatively, CF-granulated and non-granular forms of Example 10 or Example 11 (or any other insulin preparations discussed herein) preparations with oral insulin can be proportionately mixed and prepared in hard-pressed tablets so as to achieve the combined effect of a rapid onset and long-lasting hypoglycemic effect in diabetics.

Example 13

An oral insulin preparation which had a very good hypoglycemic effect on a group of diabetics was made as follows.

Chemical (lipospheric) fillers, (listed above in Example 11) were added to fresh soybean powder (to prevent trypsin, which inactivates insulin). An ovomucoid freshly prepared from egg white, according to E. Fredrikcq and H.F. Deutsch (J. Biol. Chem. 181: 499, 1949); was thus combined and dissolved with crystalline insulin in a water and alcohol mixture (50 - 50 ratio) at a pH of 7.2. Calcium chloride, 2 mm, was added to improve the physical stability of neutral insulin solution (J. Brange and S. Havelund; Properties of Insulin Solution: In Artificial Systems for Insulin Delivery, Ed. by_

P. BRUNETTI, et al., Raven Press, New York, pp. 83-88). The ovomucoid from egg white is a strong trypsin inhibitor. Alternatively, fresh egg white dissolved in alcohol can be used.

The following components are placed in the vessel for Spir-A-Flow.

These powdered components were ground by the Jet Mill into particles with a size of less than 50 microns. The particles were carefully mixed and dried in the vessel for Spir-A-Flow under the conditions described in Example 10. The components which were the dried powder were suspended in the vessel and bound with a binding solution containing insulin, ovomucoid (or egg white), and citric acid and a surfactant. The binding solution was prepared as follows:

These chemicals were dissolved in ethanol (100 - 200 ml).

These chemicals were dissolved in approx. 100 to approx. 200 ml of a phosphate buffer, with a pH of 8.0, the ethanol solution and the phosphate buffer solution, dissolved in chemicals were mixed and insulin was added. The pH of the solution was adjusted to 7.2.

The resulting binder solution was sprayed onto the fine, dried, air-suspended lipospheric powders consisting of cholesterol, surfactant, soybean powder and anti-microbial preservative as described in Example 10.

After careful drying of the insulin-binding "lipospheric" powders, these powders were coated with the following coating solution.

The above chemicals were dissolved in approx. 150 to approx. 400 ml of ethanol, sufficient ethanol to allow the chemicals to dissolve.

The above-mentioned lipid coating solution was sprayed onto the insulin-bound lipospheric powder as described in Example 12.

Test 13 clinical hypoglycaemic effect of oral insulin preparation.

Four diabetics, two men and two women, average age 46.0 (36 - 56) years old, with a weight an average of 60.3 (50 - 74) kg, and with a height of 164.3 (158 - 168) Cm, received a average of 0.573 (0.46 to 0.68) IU per kg. of the oral insulin preparation from example 13 after fasting. A marked and clinically significant degree of lowering of blood sugar levels was observed (see figure 29).

It must be clear that the various changes and modifications to the preferred designs described herein will be obvious to those skilled in the art. Such changes and modifications may be made without departing from the spirit and scope of the present invention and without diminishing its attendant advantages. It is therefore intended that such changes and changes are covered by the attached requirements.

Claims (75)

1. Preparation adapted for oral administration of a selected protein material in biologically active form, which preparation is characterized by (a) particles with an average diameter of approx. 1 to approx. 100 microns consisting mainly of a solid emulsifier and a surfactant; (b) a selected biologically active protein material bound to the surface of said particles with a binding agent; and (c) a lipid coating with a thickness of approx. 0.05 to approx. 1.0 micron surrounding said particles with said protein materials bound to their surfaces. 2. Mixture according to claim 1 further characterized by ; (d) an enteric coating surrounding said lipid coating on said particles. 3. Mixture according to claim 2 characterized in that the protein material is insulin. 4. Mixture according to claim 1 characterized in that the solid emulsifier is selected from the group consisting of cholesterol, glycerol monostearate, oleic acid, polyethylene 50 stearate, polyoxy-40 stearate, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, propylene glycol diacetate, propylene glycol monostearate, arginine HCl , gum arabic, cholesterol esters, phospholipids and fatty acids. 5. Mixture according to claim 1 characterized in that the surfactant is selected from the group consisting of sodium lauryl sulfate, sodium stearate, stearic acid, sorbitan monolaurate, sorbitan monostearate, and emulsifying wax. 6. Mixture according to claim 1 characterized in that the biologically active protein mixture is selected from the group consisting of insulin, urokinase, Factor VIII, leuprolide, gangliosides, vincristine, belomyein, lidocaine, gentamycin, bretylium tosylate, cetiedil, cyclandelate, erythromycin, chloramphenicol, adriamycin, streptokinase, and cephalosporin . 7. Mixture according to claim 1 characterized in that the binder consists of materials selected from the group consisting of sodium carboxymethyl cellulose, microcrystalline cellulose, ethyl cellulose, gelatin, hydroxypropyl methyl cellulose, methyl cellulose, povidone and hydroxypropyl cellulose. 8. Mixture according to claim 1 characterized in that the lipid coating consists of materials selected from the group consisting of polyethylene glycol fatty acid esters, glycerol phosphatides, phosphatidyl phosphates, egg yolk lecithin, oleic acid, mono-, di- and triglycerides, stearic acid, palmitate, cholesterol, cholesterol ester and tromethane. 9. Mixture according to claim 2 characterized in that the gastric juice-resistant coating consists of materials selected from the group consisting of hydroxypropylmethylcellulose phthalate, polyethylene glycol-6000 shellac, cellulose phthalate and polyvinyl acetate phthalate. 10. Mixture according to claim 1 characterized in that said particles further comprise anti-microbial agents. 11. Mixture according to claim 12 characterized in that the antimicrobial agents are selected from the group consisting of dehydroacetic acid, methylparaben, ethylparaben, phenol, phenylethyl alcohol, propylparaben, sodium benzoate, sorbic acid, thymol, thimerosal, sodium dehydroacetate, benzyl alcohol and butylparaben. 12. Mixture according to claim 1 characterized in that it comprises a lipolytic enzyme. 13. Mixture according to claim 12 characterized in that the lipolytic enzyme is selected from the group consisting of lipase, pancreatic lipase, amylase, protease, and bile salts. 14. Mixture according to claim 1 characterized in that the mixture further comprises enzyme inhibitors. 15. Mixture according to claim 14 characterized in that the enzyme inhibitors are selected from the group consisting of stearylamine, stearyl alcohol, triethylamine HC1, citric acid, lactic acid, pyrophosphate, triethanolamine, ethylamine tetraacetate, iodoacetamide, phenylhydrazine, hydroxylamine 3 and 8 hydroquinoline. 16. Mixture according to claim 1 characterized in that the mixture further comprises antifoam agents. 17. Mixture according to claim 16 characterized in that the antifoam agents are selected from the group consisting of stearyl alcohol and silicones. 18. Preparation adapted for oral administration of insulin in biologically active form, characterized in that the preparation includes; (a) particles with an average diameter from approx. 1 to approx. 100 microns consisting mainly of cholesterol, sodium lauryl sulfate and methyl and propyl parabens; (b) insulin bound to the surface of said particles with a solution consisting essentially of hydroxypropyl cellulose, sodium lauryl sulfate, triethylamine HCl and citric acid; (c) a lipid coating with a thickness of approx. 0.1 to 0.3 microns; and (d) an enteric coating layer consisting of hydroxypropylmethylcellulose phthalate, shellac and PEG-6000 of 0.1 micron thickness surrounding said polyethylene glycol monostearate coating on said particles. 19. Procedure for the production of preparations adapted for oral administration of selected proteinaceous material in biologically active form characterized by the steps: (a) formation of particles with an average diameter from ca. 1 micron to 1/2 millimeter and consisting mainly of a solid emulsifier and a surfactant; (b) binding a selected biologically active protein material to the surface of said particles with a binding agent; and (c) coating said particles with said protein materials bound to their surface with a lipid coating layer having a thickness of 0.05 to 1.0 micron. 20. Method according to claim 19 further characterized by the steps (d) coating said particles with a gastric juice-resistant coating layer surrounding said lipid coating layer. 21. Method according to claim 19 further characterized by the step (d) coating the particles with a water-soluble coating prior to applying the lipid coating. 22. The product from the method in claim 19. 23. The product from the method in claim 20. 24. The product from the method in claim 21. 25. Preparation adapted for oral administration of insulin in biologically active form characterized by ; particles with an average diameter of approx. 1 to approx. 100 microns consisting mainly of an emulsifier and a surfactant; insulin bound to the surface of the particles; a lipid coating surrounding the particles having protein materials bound to their surfaces; and a gastric juice-resistant coating layer that surrounds the lipid coating. 26. Method for oral administration of insulin characterized by; that a preparation for oral intake is given which consists of insulin bound to a surface on a solid emulsifying/dissolving aid particle, where insulin and particles are coated with a lipid coating, the particles have an average diameter of approx. 1 micron to approx. 100 microns, and the lipid coating is coated with an enteric coating. 27. Procedure for oral administration of insulin preparations characterized in that a preparation for oral intake is produced which comprises insulin bound to a surface of lipoprotein particles, insulin and particles are coated with a lipid coating, and the lipid coating is coated with a gastric juice-resistant coating. 28. Procedure according to claim 27 characterized in that it comprises the step of administering a lipolytic enzyme. 29. Procedure according to claim 27 characterized in that it comprises oral administration of at least one other capsule containing at least one lipolytic enzyme. 30. Preparation adapted for oral administration of a selected protein material in biologically active form, characterized in that this preparation comprises; (a) a lipoprotein mixture; (b) a selected biologically active protein material bound to the surface of the lipoprotein mixture "and" (c) a lipid coating surrounding said lipoprotein mixture and protein material. 31. Mixture according to claim 30 characterized in that it includes a water-soluble coating that surrounds the lipoprotein and the protein material, where the lipid coating covers the water-soluble coating. 32. Mixture according to claim 30 characterized in that the lipoprotein is a lipoprotein with a low specific gravity. 33. Mixture according to claim 30 characterized in that the lipoprotein comprises at least one amino acid. 34. Mixture according to claim 30 characterized in that it comprises a gel capsule, where lipid coating material is packed in the gel capsule. 35. Mixture according to claim 34 characterized in that the gel capsule is coated with a coating resistant to gastric juices. 36. Mixture according to claim 30 characterized in that the biologically active protein mixture is from the group consisting of insulin, urokinase, factor VIII, leuprolide, gangliosides, vincristine, belimyolin, lidocaine, gentamicin, bretylium toslate, cetiedil, cyclandelate, erythromycin, chloramphenicol, adriamycin, streptokinase, and cephalosporin. 37. Mixture according to claim 30 characterized in that it includes a lipolytic enzyme. 38. Mixture according to claim 37 characterized in that the lipolytic enzyme is selected from the group consisting of lipase, pancreatic lipase, amylase, protease and bile salts. 39. Mixture according to claim 38 characterized in that the lipolytic enzyme comprises lipase and bile salts. 40. Mixture according to claim 30 characterized in that the lipid coating comprises materials selected from the group consisting of the polyethylene glycol fatty acid ester, glycerol phosphatides, phosphatidyl phosphates, egg yolk lecithin, oleic acid, mono-, di- and tri-glycerides, stearic acid, palmitate, cholesterol, cholesterol ester and tromethane. 41. Preparation according to claim 35 characterized in that the gastric juice-resistant coating consists of materials selected from the group consisting of hydroxypropylmethylcellulose phthalate, polyethylene glycol-6000, shellac, cellulose phthalate and polyvinyl acetate phthalate. 42. Mixture according to claim 30 characterized in that said particles further comprise antimicrobial agents. 43. Mixture according to claim 42 characterized in that the antimicrobial agents are selected from the group consisting of dehydroacetic acid, methylparaben, ethylparaben, phenol, phenylethyl alcohol, propylparaben, sodium benzoate, sorbic acid, thymol, timersal, sodium dehydroacetate, benzyl alcohol and butylparaben. 44. Mixture according to claim 30 characterized in that the mixture further comprises enzyme inhibitors. 45. Mixture according to claim 44 characterized in that the enzyme inhibitors are selected from the group consisting of stearylamine, stearyl alcohol, triethylamine HC1, citric acid, lactic acid, pyrophosphate, triethanolamine, ethylamine tetraacetate, iodoacetamide, phenylhydrazine, hydroxylamine, 3- and 8-hydroquinoline. 46. Mixture according to claim 30 characterized in that the mixture further comprises anti-foaming agents. 47. Mixture according to claim 30 characterized in that the lipoprotein comprises a lipid and an amino acid. 48. Mixture according to claim 47 characterized in that the lipid is selected from the group consisting of cholesterol, oleic acid, stearic acid, sodium lauryl sulfate, lecithin, phosphatides, sodium lauryl sulfate and lecithin, palmitic acid and phosphatide phosphate choline. 49. Mixture according to claim 47 characterized in that the amino acid is selected from the group consisting of arginine, lysine, histidine and aspartic acid. 50. Mixture according to claim 46 characterized in that the anti-foam agents are selected from the group consisting of stearyl alcohol and silicones. 51. Preparation adapted for oral administration of insulin in biologically active form, characterized in that the preparation comprises (a) particles that have an average diameter from approx. 1 micron to approx. 1/2 millimeter and comprises a mixture of low specific gravity lipoprotein; (b) insulin bound to the surface of said particles with a binding agent; (c) a water-soluble coating layer for coating the particles and insulin; and (d) a lipid coating for coating the water-soluble layer, particles and insulin. 52. Preparation according to claim 51 characterized in that it comprises a lipolytic enzyme. 53. Preparation according to claim 51 which comprises a gastric juice-resistant coating layer as a coating for the lipid layer. 54. Procedure for the preparation of the preparation adapted for oral administration of selected protein material in biologically active form characterized by the steps: (a) formation of low specific gravity lipoprotein particles having an average diameter from about 1 micron to 1/2 millimeter; (b) binding a selected biologically active protein material to the surface of said particles with a binding agent; (c) coating particles and proteinaceous materials with a water-soluble coating; and (d) coating the water-soluble coated particles having said proteinaceous material bound to their surface with a lipid coating layer having a thickness of 0.05 to 1.0 microns. 55. Method according to claim 54 further characterized by the step (e) coating said particles with a gastric juice resistant coating layer surrounding said lipid coating layer. 56. Product of the method in claim 54. 57. Product of the method in claim 55. 58. Preparation adapted for oral administration of insulin in biologically active form characterized by : low specific gravity lipoprotein particles; insulin bound to the surface of the particles; a water-soluble coating surrounding the particles and insulin; a lipid coating surrounding the water-soluble coating and the particles having insulin bound to their surfaces; a gel capsule to surround the lipid coating; and a gastric juice-resistant coating layer that surrounds the gel capsule. 59. Procedure for treating diabetes characterized by the steps: oral administration of at least one capsule having a composition comprising a low specific gravity lipoprotein, insulin bound to the lipoprotein, a water-soluble coating over the insulin and lipoprotein, and a lipid coating over the water-soluble coating; and oral administration of at least 1 capsule with a composition comprising lipolytic enzymes and bile salts. 60. procedure for treating diabetes characterized by the steps: orally administering at least one capsule with a composition comprising particles of emulsifier/dissolving agent, insulin bound to the particles and a lipid coating over the particles; and oral administration of at least 1 capsule of a mixture comprising lipolytic enzymes. 61. Preparation useful for oral administration of a protein compound characterized by lipolytic enzymes selected from the group comprising pancreatic lipase, amylase, protease and bile salts. 62. Mixture according to claim 61 characterized in that the lipolytic enzymes selected are pancreatic lipase and bile salts. 63. Mixture adapted for oral administration of insulin characterized by the following components: (a) particles of pancreatic lipase; (b) particles of bile salts; (c) lipid-coated insulin-bound lipid particles; (d) sodium bicarbonate; and (e) citric acid. 64. Mixture according to claim 63 characterized by the fact that the components are filled in hard gel capsules. 65. Mixture according to claim 63 characterized in that the components are pressed into tablets. 66. Mixture according to claim 63 characterized in that the weight percentage ratio between sodium bicarbonate and citric acid is between 3:1 and 1:1. 67. Mixture according to claim 63 characterized in that the insulin-bound particles have a ball size of less than 1.5 millimetres. 68. Mixture according to claim 63 characterized in that the pancreatic lipase and bile salt particles are coated with a gastric juice resistant coating. 69. Mixture adapted for oral administration of insulin characterized by a sodium bicarbonate and citric acid mixture in a ratio of approx. 3:1 to 1:1, where the mixture forms a seed which is coated with a layer of an insulin-lipid mixture, and a resulting particle is then coated with an enteric coating. 70. Method for the production of a preparation suitable for oral administration of insulin in biologically active form characterized by the steps; formation of low specific gravity lipoprotein particles; binding of insulin to the surface of the particles; coating the particles and insulin with a water-soluble coating; coating the water-soluble coating with a lipid coating; filling lipid- and water-soluble coated insulin particles into a gel capsule; and coating the gel capsule with a gastric juice-resistant coating layer. 71. Procedure according to claim 19 characterized by the step of providing a biologically active protein mixture selected from the group consisting of insulin, urokinase, factor VIII, leuprolide, gangliosides, vincristine, belomyein, lidocaine, gentamicin, betrylium toselate, setiedil, cyclandelate, erythromycin, chloramphenicol, adriamycin, streptokinase, and cephalosporin . 72. Procedure according to claim 54 characterized by the step of making the mixture so that it comprises a lipolytic enzyme. 73. Method for producing a preparation adapted for oral administration of insulin characterized by the steps (a) formation of pancreatic lipase particles; (b) formation of bile salt particles; (c) formation of lipid particles; (d) binding of insulin to lipid particles; (e) coating the insulin-bound lipid particles with a lipid coating; (f) formation of sodium bicarbonate; (g) formation of citric acid; and filling the particles of pancreatic lipase, bile salt, sodium bicarbonate, citric acid and lipid-coated insulin-binding lipid particles in hard gel capsules. 74. Procedure in accordance with requirement 73 characterized by the components being pressed into tablets instead of being filled in gel capsules. 75. Procedure in accordance with requirement 73 characterized in that sodium bicarbonate and citric acid are used in a weight percentage ratio between sodium bicarbonate and citric acid between 3:1 and 1:1. SUMMARY A composition adapted to administer a pharmacologically effective amount of a pharmaceutical agent when taken orally, comprising (a) a pharmaceutical agent; (b) a lipid coating material and (c) a gastric juice resistant coating material and optionally comprising water soluble coating materials; emulsifying and dissolving aids; binders; stabilizers and enzyme inhibitors, surfactants and antimicrobials. The mixture may also comprise a low specific gravity lipoprotein mixture and lipolytic enzymes.