JPH03170438A - Biologically active protein composition for oral and local administration - Google Patents

Abstract

PURPOSE:To obtain the subject composition, containing a biologically active lecithoprotein, having a high absorption rate and also the high maximum absorbed concentration and capable of sustaining the concentration in the body for a long period. CONSTITUTION:A composition containing a biologically active lecithoprotein alone or together with a proteolytic enzyme inhibitor (e.g. aprotinin, urinastatin or soybean protease inhibitor) without inhibiting the aforementioned biologically active protein. Insulin, calcitonin, interferons, kallidinogenase, superoxide dismutase, elastase, immunological immunopeptides, drugs having a protein in a substituent group, etc., are cited as the biologically active protein. The above-mentioned protein which can be conventionally administered merely by injection can be used through routes of oral and local administration.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to bioactive protein compositions for oral and topical administration. [Prior Art] With the exception of a few, biologically active proteins such as calcitonin, insulin, interferons, and immune immunopeptides are administered parenterally, and are generally administered orally and locally. do not have. It would be very advantageous from various points of view if administration could also be carried out orally and locally. [Summary of the Invention] The present inventor has discovered that by first converting a biologically active protein into lecithin, excellent effects such as enhancing the pharmacological activity of the biologically active protein and reducing side effects can be achieved. Furthermore, it has been found that oral and local administration is possible when this lecithinized bioactive protein is used alone or in combination with a protease inhibitor that does not inhibit the bioactive protein. That is, the present invention relates to bioactive protein compositions for oral and topical administration comprising lecithinated bioactive protein alone or a protease inhibitor that does not inhibit the bioactive protein. The biologically active protein used in the present invention may be any protein as long as it has biological activity. For example, insulin, calcitonin, interferons, kallidinogenase, superoxide dismutase, elastase,
Examples include, but are not limited to, urokinase, immunoimmunobutide, and drugs having protein substituents.
In the present invention, lecithinized products of these biologically active proteins are used. In the lecithinized bioactive protein, the bioactive protein is bound to lecithin of the following formula (1) through chemical crosslinking. CHzOCO(CHi). CHs 1 (wherein n is an integer from 1 to 20, m is an integer from 1 to 20, and 2 is -Nut or -GOOR) formula (
In the compound of 1), m is preferably an integer of 3 to 20, most preferably an integer of 14 to 20. The alkyl group in which two groups are bonded may be optionally substituted with one or more alkyl groups having 1 to 4 carbon atoms. Furthermore, in order to stabilize the compound, a compound in which two groups are converted to t-Boc may be used. Examples of methods for chemical crosslinking include the following. In the compound of formula (1), the Z group is one COOH
When the frequency is -MHz, for example, the carbodiimide method and the cyanuric chloride method are used, and when the frequency is -MHz, for example, SPDP[N-
Succinimidyl 3-(2-pyridyldithio)propionate] method, carbodiimide method, and cyanuric chloride method. In the carbodiimide method, for example, l-ethyl-3-(dimethylaminoprobyl)carbodiimide (EDC) or dicyclohexylcarbodiimide (
DCC), the biologically active protein and the compound of formula (1) are bonded through a -CONH bond. In addition, in the cyanuric chloride method, cyanuric chloride is used and the two are bonded together through a bond. Furthermore, in the SPDP method, −N
H-Co-CHtCHt-S-S-CHzC}licO
Both are bonded by an NH bond. In these methods,
The reaction is carried out under conventional reaction conditions or conditions close to them.

One embodiment of the composition of the present invention includes, in addition to the lecithinated biologically active protein, a protease inhibitor that does not inhibit the biologically active protein. These protease inhibitors include a variety of natural and synthetic compounds, including the following: Abrotinin, ulinastatin, hirudin, soybean protease inhibitor
Lima bean protease inhibitors, corn protease inhibitors, gabexate mesylate, camostat mesylate, nafamostat mesylate, etc. These inhibitors are well known and can be briefly explained as follows. Abrotinin is a basic protein with a molecular weight of approximately 6,500 extracted from cow pancreas and lungs, ulinastatin is a glycoprotein with a molecular weight of approximately 67,000 extracted from human urine, and hirudin is extracted from leech salivary glands. An acidic protein with a molecular weight of about 7000?
Soybean protease inhibitor is a protein with a molecular weight of about 22,000 extracted from soybeans, and lima bean protease inhibitor is a protein with a molecular weight of about 9,000 extracted from lima beans.
+JzJsOtS) is a white crystal or crystalline powder with a melting point of 90-93°C,
i■HiJJaS) is an odorless white crystal or crystalline powder with a melting point of 150-155°C, and nafamostat mesylate is a white crystalline powder with a melting point of about 260°C (decomposed). Some of these inhibitors specifically inhibit the biologically active proteins used in the present invention, so
It cannot be used with such biologically active proteins. For example, nafamostat mesylate strongly and selectively inhibits trypsin, kallidinogenase, and thrombin and cannot be used with these proteins. However, nafamostat mesylate does not inhibit insulin and can be used together with insulin. This can be easily determined by a person skilled in the art. In the present invention, the effect of the present invention can be further enhanced by containing such an inhibitor. The compositions of the present invention are administered in common oral and topical forms. In the present invention, local administration means transdermal, intraoral, nasal, rectal, and intrauterine administration. Its forms include, for example, tablets, capsules, powders, granules,
Examples include troches, suppositories, ointments, creams or lotions, gels, spray aerosols, and the like. For oral administration, the compound must be in enteric-coated form.
These dosage forms can be manufactured by methods well known to those skilled in the art. Taking tablets, capsules, for example, conventional excipients such as binders such as sugar syrup, gum arabic, gelatin, sorbitol, gum tragacanth or polyvinylpyrrolidone; fillers such as lactose, sugar, maize starch, calcium phosphate, sorbitol or Glycine; lubricant for tableting, such as magnesium stearate; disintegrant, such as starch, polyvinylpyrrolidone,
It can be produced by operations such as blending, filling, tabletting, enteric coating, etc. using sodium starch glycolate or microcrystalline cellulose; or a wetting agent such as sodium lauryl sulfate. The composition of the present invention contains an effective amount of 0.1 to 99% by weight. The compositions of the present invention can be used to treat diseases for which the biologically active proteins they contain are effective. For example, insulin
It is used to treat diabetes for which insulin therapy is indicated.
Interferon α is used to treat renal cancer and multiple myeloma, calcitonin is used to improve pain and bone loss in osteoporosis, hypercalcemia, and Paget's disease of the bone, and kallidinogenase is used to treat kidney cancer and multiple myeloma. , mainly used to treat circulatory disorders. The dosage of the composition of the present invention varies depending on the severity of the disease, the weight of the patient, etc., but is generally an amount at which each biologically active protein contained in the composition is effective. For example, for insulin, the dose is 4 to 60 units at a time, and 200 units or less per day. For synthetic inhibitors, the amount of protease inhibitors is generally 10 to 100 mg per dose and 20 to 1000 Ing per day.
is preferable. Administration is preferably carried out one or more times a day, for example several times a day. The compound of the present invention is not toxic within the above-mentioned dosage range.

The composition of the present invention allows the use of oral and topical administration routes for biologically active proteins, which could previously only be administered by injection. When administered, the absorption rate is fast and the maximum absorption concentration is also high. Furthermore, it has become possible to achieve excellent effects by maintaining the concentration in the body over a long period of time. In addition, even for only a small portion of biologically active proteins that can be administered orally or locally, the composition of the present invention has a faster absorption rate, a higher maximum absorption concentration, and a longer concentration in the body than conventional products. It has the advantage of being long-lasting. These advantages are not available with conventional products.

[Example] Next, examples will be shown.

Example 1 Aminolecithin of formula (1) (in the formula, Z is t-Bo
cNH, n is 5) 30 mg (0.04 mm
BOC L/, TFA
After completely distilling off, it was dissolved in distilled water and neutralized with Na2COa. Insulin 5mg (0. O O
1-ethyl 3-(3-dimethylaminobrobyl)carbodiimide (EDC) was added to a final concentration of 0.1M, and the mixture was stirred overnight at room temperature. A rat (Wister species, male, 250 g) was opened and the duodenum was injected with 30 or 100 g of lecithinated insulin.
Units/kg and a mixture of nafamostat mesylate 10-3H were administered. Separately, as a control, a mixture of 100 units/kg of insulin and 10-3M of nafamostat mesylate was administered. Administration 0, 301 60
, 901 Blood sugar levels were measured after 120 and 180 minutes.
The results are shown in Table 1. Everyone go 0 61 60
6530 116 101
11380 112
57 11490 8
9 85 105120
83 84
111180 84 79
119 As can be seen from Table 1, the blood sugar level of the control hardly changes and no effect is observed, but with the composition of the present invention, a sufficiently excellent effect is observed even at one-third the amount of the control. ．． Furthermore, the effects can be sustained over a long period of time. Example 2 Lecithinated kallidinogenase was obtained in the same manner as in Example 1, using 5 μg of aminolecithin of formula (1) used in Example 1 and 250 μg of [ ”H]-kallidinogenase. In the same manner as in 1, [3H] lecithinated kallidinogenase 60 μg/10 μl was administered to rats at a concentration of 200 μg/10 μl per rat.
μl was administered. In addition, [3H] kallidinogenase was similarly administered as a control. The concentration of [3H] kallidinogenase in the blood was determined by administration 30,
Measurements were taken after 60, 90, 120 and 180 minutes. The results are shown in Table 2. Table 2 30 2. 20
0. 5060 2. 68
1. 3290
2.81 2.06120
1. 98 1. 68180
1. 48 1.
35 Table 2 shows that kallidinogenase is conventionally administered orally as an enteric preparation in addition to an injection, but the composition of the present invention has a fast absorption rate, a high maximum absorption concentration, and a long-lasting concentration in the body. It turns out that there are advantages.

Claims (2)

[Claims] (1) A bioactive protein composition for oral and topical administration containing a lecithinized bioactive protein. (2) A bioactive protein composition for oral and topical administration, comprising a lecithinized bioactive protein and a protease inhibitor that does not inhibit the bioactive protein.