DE3018843A1 - INSULIN PREPARATION AND METHOD FOR PRODUCING THE SAME - Google Patents

Description

Insulin preparation and process for making the same

The invention relates to a new insulin preparation and more particularly to an insulin preparation for the treatment of diabetes and a method for producing the same. The preparation according to the invention comprises a carrier for preparations for absorption through the mucous membrane, insulin and a special acid.

Insulin is widely used to treat diabetes. Since insulin is a polypeptide, the Proteinase in the digestive fluid inactivated if administered orally. It is therefore general Common practice of injecting insulin intravascularly or intramuscularly. These insulin injections require substantial exercise and they also cause a variety of ailments. Attempts have therefore been made to avoid the injection to be replaced by other methods of administration. Into the-

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special preparations were developed for oral administration as well as Suppoaltorlen for rectal or vaginal administration of the insulin.

Recently, methods have attracted particular attention _t in which the insulin is absorbed through the mucous membrane and not through the digestive tract. It is of particular interest to create insulin preparations in the form of suppositories. With this form of administration, no special countermeasures have to be taken against inactivation of the insulin by proteinase. However, insulin is not satisfactorily absorbed from the intestinal tract when it is only in a substrate for suppositories. Additives to facilitate the absorption of insulin have been investigated and it has been proposed, for example, to use a surface-active agent together with insulin [Nishioka et al, "Pharmacology ¹¹ Nihon Yakugaku Kai, 2 £, 88 and 119 (1977) j JA-OSen 52 -87218, 53-6417, 53-18723 and 53-107408], an agent containing a phosphatide, bile acid or the like. Together with insulin (JA-OS 52-83923) and an agent containing a proteinase inhibitor together with insulin (JA-OS 53-15412). However, these known preparations require a large amount of insulin to ensure a sufficiently low blood sugar level and are therefore not used in practice.

Intensive studies have been carried out to overcome the difficulties mentioned. In the course of this Studies have shown that inaulin is excellently absorbed through the mucous membranes when it is in shape of a special preparation is available. This essentially consists of insulin, an acid and a carrier for preparations that are used for mucosal resorption are determined. In addition, you can also have a

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Proteinase inhibitor provide a long-term effect of insulin. This effect is especially noticeable when the preparation is in the form of suppositories for rectal or vaginal administration or in the form of a sublingual tablet. One achieves one significant decrease in blood sugar level, even if the insulin is only present in very small amounts, e.g. on the order of one to several tenths of the amount of insulin in conventional suppositories.

The specific acid of the preparation according to the invention is selected from the group of the following acids. It can only one acid can be selected or multiple acids can be selected.

(1) lower saturated fatty acids having 1 to 6 carbon atoms;

(2) fatty lower hydroxycarboxylic acids;

(3) inorganic acids, selected from the group consisting of hydrochloric acid, phosphoric acid, phosphotungstic acid and perchloric acid}

(4) aromatic carboxylic acids from the group consisting of benzoic acid, phthalic acid and pyromellitic acid;

(5) substituted acetic acids from the group monochloroacetic acid, dichloroacetic acid, trichloroacetic acid and suIfoeaic acid;

(6) unsaturated carboxylic acids from the group consisting of acrylic acid and maleic acid; and

(7) pyruvic acid, furoic acid and picric acid.

The first type of acid is saturated lower fatty acids with 1 to 6 carbon atoms, in particular Formic acid, acetic acid, propionic acid, butyric acid, valeric acid and caproic acid. Saturated are preferred lower fatty acids with 1 to 5 carbon atoms. From a commercial point of view, they are saturated

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lower fatty acids with 1 to 3 carbon atoms preferred, i.e. formic acid, acetic acid and propionic acid.

The second type of acid concerns lower fatty hydroxycarboxylic acids, such as tartaric acid, malic acid, citric acid, lactic acid, Glycolic acid or the like. Fatty hydroxycarboxylic acids of the type D-olucoronic acid and L-ascorbic acid (vitamin C), i.e., generally carboxylic acids derived from sugars are ineffective.

The third type of acid is inorganic acids such as hydrochloric acid, phosphoric acid, nitric acid, phosphotungstic acid (P ₂ O ₅ .24WO ₅ .nH ₂ 0) and perchloric acid. Other inorganic acids, such as sulfuric acid or perbromic acid, have no effect.

The fourth acid group is aromatic carboxylic acids and in particular benzoic acid and phthalic acid and pyromellitic acid.

The fifth group of acids are substituted acetic acids, such as monochloroacetic acid, dichloroacetic acid, trichloroacetic acid and sulfoacetic acid (HO ₃ S-CH ₂ COOH).

The sixth group is unsaturated carboxylic acids such as acrylic acid and maleic acid.

The seventh group includes pyruvic acid, furoic acid and Plcrinic acid.

The desired effect can only be achieved if the acids mentioned are selected. Amino acids or saturated lower ones Fatty acids with amino groups, such as glutamic acid, phenylalanine, aspartic acid, γ-amino-n-butyric acid, 6-amino-n-

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caproic acid or the like., show only a low effectiveness. In practice it is necessary to use the desired additional compounds in the form of the free acids. They are not effective in salt form.

All of the acids described above are suitable for achieving the aim of the invention. However, the following acids are preferred for their effectiveness: (1) lower fatty acids having 1 to 5 carbon atoms; (2) tartaric acid and glycolic acid; (3) hydrochloric acid and nitric acid; and (4) monochloroacetic acid. Among these deliver the lower fatty acids, in turn, give the best results. Formic acid has an irritating effect. on the other hand higher fatty acids above propionic acid show an unpleasant odor. Therefore, acetic acid is particularly preferred.

It cannot yet be explained how these acids promote the absorption of insulin. It will, however assumed that the mechanism of action is not only based on a lowering of the pH value, as with the subsequent experiments can be shown.

The acids can be added to the preparations in the respectively effective amount in pure form or in the form of an aqueous solution can be added. If the amount of acid is too small, there remains a notable effect on the absorption of the Insulin. If the amounts are too large, the acid has too strong an effect on the mucous membranes. Hence there is a concentration of the acid in the total preparation in the range from 0.01 to 0.5% by weight and preferably 0.02 to 0.1% by weight.

If you also provide a proteinase inhibitor, so can maintain the effect on increasing the absorption of insulin through the mucous membranes over a longer period of time will.

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Han can use all common proteinase inhibitors, as long as they only fulfill the desired function. In particular, all inhibitors are suitable, which of Natural substances are derived, in particular inhibitors of animal or vegetable origin. There come various synthetic materials can also be used as proteinase inhibitors. A natural proteinase inhibitor can be obtained from beef lungs; the synthetic proteinase inhibitors are preferably low molecular weight ones sold under the trademark FOY (Ono Pharmaceutical Co., Ltd.) are sold and the main ingredient is dimethylcarbamoylmethyl-p- (p-guanidinobenzoyloxy) -phenylacetat.Mesitylat contain.

The source of insulin is not critical. Any type of insulin can be used as long as it is only used for lowering the blood sugar level are suitable, e.g. B. bovine insulin, pig insulin, whale insulin or the like., Or but also synthetic insulin. The insulin is used in an effective amount of generally 0.1 to 100 I.U. and preferably 0.5 to 50 IU / g of the preparation.

The insulin preparation according to the invention can be used in the conventional manner in the field of preparations for mucosal resorption Way to be made.

The insulin and the acid can be added separately to the carrier for the preparation for mucosal resorption. However, it is preferred to first dissolve the insulin in an aqueous solution of the acid and then this Add the solution to the carrier. In this case it is it is particularly preferred to melt the carrier material prior to mixing. The improvement of insulin absorption

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tion is particularly pronounced if the insulin is only added to the preparation after it has been in an aqueous solution the acid was dissolved. Therefore, this mode of operation is in contrast to a separate incorporation of the insulin and the acid is preferred.

If insulin is previously dissolved in an aqueous solution of an acid, care must be taken that the total added amount of water is small enough to keep the final preparation in a solid state.

The expression "preparation for mucosal resorption ¹¹ denotes solid preparations for parenteral mucosal resorption and in particular suppositories which are suitable for rectal or vaginal use, as well as sub-lingual tablets. The expression" carrier for a preparation for mucosal resorption ¹¹ denotes a carrier or excipient, which is commonly used in the manufacture of solid preparations for parenteral administration and mucosal resorption.

As an excipient for sub-lingual tablets, all excipients commonly used in this field can be used Question, e.g. sorbitol, mannitol, lactose, glucose, sucrose, glycerin, crystalline cellulose, gum arabic or a Mixture of the same.

Lipophilic or hydrophilic suppository auxiliaries are suitable as excipients for suppositories. As lipophilic Suppository clay excipients come individually or in combination natural oils and fats in question, such as coconut oil, peanut oil, olive oil, soybean oil, rapeseed oil, cottonseed oil, Sesame oil, corn oil, rice bran oil, camellia oil, cocoa butter oil, lard, wool fat, beef tallow or the like. There are also hydrogenated products made from it,

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-β -

acetylated products and extracts in question. Synthetic esters of higher fatty acids and glycerin can also be used Question or of higher fatty acids and alcohols with 2 to β carbon atoms.

As hydrophilic aid materials for suppositories are PoIyäthylenglykol propylene glycol _f Glocellogelatine or the like. In question. If a lipophilic suppository excipient is used as an excipient and the insulin is previously dissolved in an aqueous acid solution and then incorporated into the substrate, the aqueous acid solution with the insulin dissolved therein is in the form of fine aqueous droplets in the substrate, so that a relatively small amount of acid is sufficient to achieve the desired effect.

In addition to the essential components mentioned, the preparation according to the invention can contain a number of other components Contain additives that are usually included in preparations of this class. These additives can are used in amounts which do not interfere with the effect intended by the present invention.

A surface-active agent can be added to the preparations according to the invention in the form of suppositories to add. Nonionic, cationic, anionic and amphoteric surfactants are suitable. Examples of the nonionic surfactants are monoglycerides or diglycerides of higher fatty acids, sorbitol anhydride fatty acid ester, polyoxyethylene sorbitol anhydride fatty acid ester, Polyoxyethylene fatty acid ester, polyoxyethylene higher alcohol ether, Polyoxyethylene-alkylphenyl ether, castor oil-ethylene oxide adducts, saccharode fatty acid esters or the like. * Quaternary ammonium salts are suitable as cationic surfactants, e.g.

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-Sf-

long chain monoalkyl ammonium halide and short chain Trialkylammonium halides, long-chain dialkylammonium halides and short chain dialkyl ammonium halides, long chain alkyl benzyl ammonium halides and short-chain alkylbenzyl-ammonium halides, long-chain Alkyl pyridinium halides, benzethonium halides or the like. A chloride or a bromide is particularly suitable as the halide. Examples of the anionic Surface-active agents are fatty acid salts (soaps), Alkyl sulfates, alkylbenzenesulfonates, polyoxyethylene alkyl ether sulfates, Alkyl sulfosuccinates, N-acylamino acid salts, Alkyl phosphates, polyoxyethylene alkyl ether phosphates or the like .. Suitable examples of the counterion of the Surfactants are alkali metals, such as sodium, potassium or the like., Alkanolamine salts, such as monoethanolamine, Diethanolamine, triethanolamine or the like. The anionic surfactant can be in the form of the free Acid or in the form of a salt can be used. Any amphoteric surfactants can be used Question, e.g. agents of the betaine type, of the sulfobetaine type or of the type of basic amino acids with an N-acyl group. Lecithin can also be used.

Other additives for suppositories are bile acids, in particular cholic acid, taurocholic acid, glycolic acid, deoxycholic acid, taurodeoxycholic acid, glycodeoxycholic acid, Chenodeoxycholic acid, taurochenodeoxycholic acid, glycochenodeoxycholic acid and their alkali metal salts with Sodium, potassium or the like.

The insulin preparation according to the invention is suitable for rectal or vaginal administration, if it is in the form of In suppositories or for administration in the form of sub-lingual tablets. It is extremely effective for Lowering blood sugar levels even when the insulin

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is only present in small amounts of one to several tenths of the insulin content in conventional suppositories.

In the following the invention is illustrated by means of examples explained in more detail.

Experimental example 1

Male rabbits, each weighing 2.5 kg, are kept without food for 24 hours and then laid on their backs. A solution shown in each case in Table 1 is administered rectally to each rabbit at a depth of about 2.5 cm from the anus. A cannula is then inserted into the femoral vein of the rear leg of each rabbit in order to withdraw about 0.2 ml of blood at certain time intervals. The blood sugar value is determined by the oxygen method [Kanal et al., "Handbook of the Clinical Inspection ¹ *, 25th Ed., Kanehara Shuppan, VII-51 (1968)] and the change in the blood sugar value over time is determined before treatment it is assumed with 100 # The results are shown in Table 1.

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Approved solution

Table 1

Percentage change based on the blood sugar level before administration

15th
min

30th
min

45
min

60
min

90 min

comparison

physiological saline solution 1 g insulin 0.7 IU / kg

aqueous 0.6 # acetic acid solution physiological salt solution 0.9 g FOY ⁺ 0.1 g

Hw 0.7 IU / kg

+9.1 +15.2 +12.1
+0.5 +11.4 +2.9

-8.0 -12.0

+12.1 +4.5

-15.0

+21.2 +2.8

-13.8

^ο present invention ~ j aqueous 0.3 # acetic acid solution 1 g ■ ^ insulin 0.7 IU / kg

aqueous 0.6 # acetic acid solution 1 g insulin 0.7 I.U./kg

aqueous 3% acetic acid solution 1 g insulin 0.7 IU / kg

-12.0 -25.5 -20.0 -23.3 -20.0 -33.4 -33.7 -20.0 -15.2 -31.2 -50.9 -33.0

-26.7 -16.2 -8.0 -18.2

120

min

+27.0 +7.2

-6.7

-27.6 -13.3 -3.6 -18.2

aqueous 0.6 # acetic acid solution 0.9 g _ _18f2 _ _13f3 " _34f5 ^ ₃
Insulin 0.7 IU / kg

⁺ Trademark for dimethylcarbamoylmethyl-p- (p-guanidinobenzoyloxy) -phenyl acetate.Mesitylate,
a synthetic low molecular weight proteinase inhibitor manufactured by
Ono Pharmaceutical Co., Ltd.

The values in the table are based on a 1OOJ6 assessed blood glucose level before administration. If the blood sugar level increases, there is a "+" sign preceded by a "-" sign if the blood sugar level has fallen.

The results according to Table 1 can be interpreted as follows will. If the physiological saline solution and insulin or the aqueous acetic acid solution for comparison are administered, the blood sugar level rises above the level determined before the treatment. This is on the Attributed to the stress that rabbits are subject to during treatment. It is a common phenomenon. The results of the experiments according to the invention show that in the case of conventional rectal administration a An amount of insulin in the range of several IU / kg to several 10 IU / kg is required to achieve a significant reduction to achieve the blood sugar level (above about 15%), while, on the other hand, the preparation according to the invention has a significant effect on the blood sugar level even at one shows a small amount of only 0.7 IU / kg. It can also be seen that when using a proteinase inhibitor together with acetic acid, the desired effect is maintained for a longer period of time.

Experimental example 2

Nan carries out a similar test as in Test Example 1. However, there are now various acids in terms of investigated their effect on lowering blood sugar levels. The results are shown in Table 2. In all cases, insulin is used in an amount of 0.7 IU / kg. The various, in Table 2 The acids indicated are used in such an amount that 1 g of an aqueous 0.1N solution is obtained.

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Table 2

acid Percentage change in blood sugar
value, based on the value before the
Treatment X%) 40 min 60 min 120 min 20 min according to the invention -44.4 -33.4 -16.7 Formic acid -31.5 -40.9 -44.6 -13.6 Propionic acid -31.9 -27.8 -38.9 -43.4 Butyric acid -13.9 -40.8 -37.5 +5.3 Valeric acid -21.1 -14.3 +3.6 -2.4 Caproic acid -7.1 -17.9 -10.7 0 Tartaric acid -7.1 -19.1 -24.3 +10.8 Malic acid -8.9 -32.2 -33.4 -40.8 Glycolic acid -23.7 -31.9 -18.2 -9.1 hydrochloric acid -27.3 -29.6 -26.2 -18.5 Phthalic acid -26.2 -36.8 -37.6 -38.4 Monochloroacetic acid -24.3 -27.0 -29.2 -11.6 Acrylic acid -7.7 -19.0 -34.8 -9.5 Pyruvic acid -4.8 comparison +14.3 +7.1 - Adipic acid +21.4 0 +21.7 - Fumaric acid +13.0 +11.1 +10.6 +10.9 sulfuric acid 0 +35.0 +30.0 +22.1 Glutamic acid +30.0 +25.6 +28.2 +24.3 D-phenylalanine -3.8 +16.3 0 - D-glucuronic acid +6.3

It can be seen from Table 2 that the incorporation of specific acids in the sense of a stronger lowering of the blood sugar value works while other acids do not give these results.

Experimental example 3

An experiment similar to that in experimental example 1 is carried out carried out. Insulin (0.7 IU / kg) is dissolved in a pH buffer solution and the change in blood sugar

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value is determined after administration of this solution. The results are shown in Table 3.

Table 3

PH Buffer solution Change in blood sugar level,
based on the value before the
abreichun " (%) 40 min 50 min 20 min 0 +5.5 2.0 Glycine hydrochloric acid +4.5 -3.6 0 4.0 Acetic acid-NaOH -7.1 +13.3 +16.7 7.0 Tris (hydroxymethyl) -
aminomethane hydrochloric acid +20.0 +12.7 +27.5 9.0 dito +32.4 +15.8 +26.3 11.0 Glycine-NaOH +15.8

It can be seen from the results in Table 3 that the effect of the acid on the acceleration of the absorption or absorption of the insulin is not based solely on a lowering of the pH value.

9 g of isocacao (trademark for higher fatty acid triglycerides, manufactured by Kao Soap Co., Ltd.) are added at 42 to 43 ° C and then treated with 20 IU insulin, dissolved in 1 g of a 0.6 # acetic acid solution. then is stirred sufficiently to make a dispersion. Hit this dispersion with each suppository weighing 1 g in a mold.

The suppositories are administered to male rabbits weighing about 3 kg, which in the course of the received no food for the previous 24 h. and on the Backs were set. The insulin in the plasma is then quantitatively determined by radio-immunoassay methods, using a two antibody method. A preparation is used for comparison in its preparation instead of the above acetic acid solution a physiological saline solution was used. The results are shown in Table 4.

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Table 4

Concentration of the insulin in the preparation

Plasma (/ uE / ml)

O 20 405Ö90 TH) 180 min min min min min min min

comparison

physiological salt solution 0.1 g '

Insulin 0.7 IU / kg 4 10 16 16 15 15 14 isocacao 0.9 g

invention

0.6 # acetic acid solution 0.1 g

Insulin 0.7 IU / kg 8 336 290 118 92 68 31 Isocacao 0.9 g

example 1

10 g Isocacao (tradename for higher fatty acid glyceride of the Kao Soap Co., Ltd.) are melted at 42 to 43 ⁰ C. Then 20 IU of insulin are dissolved in 0.1 ml of an aqueous oxygenated acetic acid solution and this solution is added to the melt with stirring and dispersion. 1 g of the dispersion is placed in a suppository mold and solidified and then removed.

Example 2

Example 1 is repeated, 0.1 ml of an aqueous 4.5 # strength formic acid solution being used. Receives one insulin suppository.

Example 3

Example 1 is repeated, 0.1 ml of an aqueous 8.5 # propionic acid solution used.

Example 4

Example 1 is repeated using 0.1 ml of an aqueous T5 # tartaric acid solution.

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example

Example 1 is repeated using 0.1 ml of an aqueous 3 »5 strength hydrochloric acid solution.

Example 6

9 g isocacao (of the type specified in Example 1) are melted at 42 to 43 ° C and this melt is with 20 IU insulin, dissolved in a 6 # aqueous acetic acid solution, added as well as 1 g of a fine powder from FOY (proteinase inhibitor with low molecular weight according to test example 1). The mixture is sufficiently stirred and dispersed. 1 g of the mixture is filled into a suppository mold and solidified.

Example 7

To 9.9 g Isocacao (as in Example 1), 0.1 g PoIyoxyäthylen-sorbitanhydridfensäureester (Tween 80, trade name of Kao Atlas Co., Ltd.) are and the mixture is melted at 42 to 43 ⁰ C. 20 IU of insulin, dissolved in 0.1 ml of an aqueous 6% acetic acid solution, are added to the melt. Thereafter, sufficient agitation is used to produce a dispersion. 1 g of the mixture is poured into a suppository mold and made to solidify.

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Claims (14)

30188A3 - vT claims 1. Insulin preparation, consisting essentially of an excipient for preparations for mucosal resorption, Insulin and 0.01 to 0.5 percent by weight, based on the total preparation, of one or more acids selected from following group: (1) lower saturated fatty acids having 1 to 6 carbon atoms ; (2) fatty lower hydroxycarboxylic acids; (3) an inorganic acid selected from the group consisting of hydrochloric acid, phosphoric acid, nitric acid, phosphotungstic acid and perchloric acid; (4) an aromatic carboxylic acid selected from the group consisting of benzoic acid, phthalic acid and pyromellitic acid; (5) a substituted acetic acid selected from the group consisting of monochloroacetic acid, dichloroacetic acid, Trichloroacetic acid and sulfoacetic acid; (6) an unsaturated carboxylic acid selected from the group consisting of acrylic acid and maleic acid; and (7) pyruvic acid, furoic acid and picric acid. 2. Insulin preparation according to claim 1, characterized in that that the acid is a lower saturated fatty acid having 1 to 5 carbon atoms. 3. insulin preparation according to claim 2, characterized in that the acid is a lower saturated fatty acid having 1 to 3 carbon atoms. 4. insulin preparation according to claim 1, characterized in that that the acid is one or more lower fatty hydroxycarboxylic acids selected from the group of tartaric acid, Malic acid, citric acid, lactic acid or glycolic acid. 030047/0947 . ordinal inspected 5. insulin preparation according to claim 4, characterized in that the acid is tartaric acid or glycolic acid. 6. insulin preparation according to claim 1, characterized in that the acid is hydrochloric acid or monochloroacetic acid is. 7. insulin preparation according to claim 1, characterized in that the acid is monochloroacetic acid. 8. insulin preparation according to claim 1, characterized in that the excipient for preparations for mucosal absorption is an excipient for suppositories. 9. insulin preparation according to claim 1, characterized in that it also contains a surface-active agent contains. 10. Insulin preparation according to one of claims 1 to 9, characterized by a content of a proteinase inhibitor. 11. insulin preparation according to claim 10, characterized in that the acid is a lower saturated fatty acid having 1 to 5 carbon atoms. 12. insulin preparation according to claim 10, characterized in that the acid is one or more lower fatty hydroxycarboxylic acids, selected from the group of tartaric acid, malic acid, citric acid, lactic acid or glycolic acid, is. 13. Insulin preparation according to claim 10, characterized by the content of a surface-active agent. 030047/0947 14. Method 2 for the production of an insulin preparation, characterized by the following steps: (I) dissolving insulin in an aqueous solution of one or more acids selected from the group (1) lower saturated fatty acids having 1 to 6 carbon atoms; (2) fatty lower hydroxycarboxylic acids; (3) an inorganic acid selected from the group consisting of hydrochloric acid, phosphoric acid, nitric acid, phosphotungstic acid and perchloric acid; (4) substituted acetic acid selected from the group consisting of monochloroacetic acid, dichloroacetic acid and trichloroacetic acid and sulfacetic acid; (5) aromatic carboxylic acids selected from the group consisting of benzoic acid, phthalic acid and pyromellitic acid; (6) unsaturated carboxylic acids selected from the group consisting of acrylic acid and maleic acid; and (7) pyruvic acid, furoic acid, and pieric acid; and (II) incorporating said aqueous insulin solution in an adjuvant for preparations for Schleimhautresorption, wherein a preparation containing 0.01 to 0.5 wt.% Of the acid, based on the total preparation, is obtained. 15 · The method according to claim 14, characterized in that the auxiliary substance before the incorporation of the insulin solution is melted. 030047/0947