JPH01233230A - Percutaneous absorption promoter and nasal drop containing the same - Google Patents

Abstract

PURPOSE:To obtain a percutaneous absorption promoter for slightly absorbable peptide hormones including insulin and glucagon, by using, as an active ingredient, glycyrrhizic acid and its derivative and their medically permissible salts. CONSTITUTION:Glycyrrhizic acid of formula I (R is Ha HOOCCH2CO, formula II), its derivative and their medically permissible salt, is used as an active ingredient to prepare a percutaneous absorption promoter for hard absorbable substance such as high molecular weight peptide hormones, for example, insulin or glucagon. The promoter can be applied in the form of an aqueous solution together containing a dose amount of slightly absorbable substance by rhinenchysis or spraying. In case of insulin, nasal administration results in the same level of satisfactory absorption as in parenteral injection.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a transmucosal absorption enhancer for poorly absorbed substances containing glycyrrhetinic acid, derivatives thereof, and pharmaceutically acceptable salts thereof as active ingredients.

[Prior Art and Problems] Among currently used pharmaceuticals, poorly absorbed drugs such as peptide hormones, such as insulin, could only be administered to diabetic patients by injection.

However, the injection method of these poorly absorbed substances, such as insulin, has the disadvantage that it causes pain during injection, and that continuous administration over a long period of time may cause tissue thickening at the injection site.

On the other hand, research has been completed on nasal sprays and layered preparations that avoid these situations and enable home administration, but insulin, for example, has a molecular weight of 6.000, It is extremely difficult to absorb through the nasal mucosa or rectal mucosa.

Therefore, in recent years, there has been no investigation into substances that promote the absorption of insulin, etc., and substances with surfactant effects such as bile acids, medium-chain fatty acids, nonionic or ionic synthetic surfactants, and saponins have been shown to have absorption-promoting effects. It has come to be known that there is.

Under these circumstances, the present application U114 etc. has investigated whether the active ingredients of licorice, such as G1,1 tyrrhizin, glycyrrhetinic acid, and their tW) MZ forms and salts, have transmucosal absorption promoting effects on insulin, glucaco, etc. We considered whether These licorice ingredients have surfactant effects,
These are used in a wide range of applications, including internal medicines, external medicines such as eye drops and ointments, and additions to cosmetics and foods as sweeteners.

As a result of various studies, the inventors of the present application have discovered that the above compound exhibits a remarkable effect on promoting transmucosal absorption of high molecular weight peptide hormones including insulin and glucagon, and has completed the present invention. It is.

Compounds useful in the present invention as those that promote absorption have the general formula (1): [wherein R is H-, HOOCCII CIl C
Glycyrrhetinic acid represented by 0-5, its derivatives, and its pharmaceutically acceptable salts.

The compounds included in the above general formula (1) are glycyrrhetinic acid, glycyrrhetinic acid 3β-3-〇-hemisuccinate, glycyrrhetinic acid 3β-3-〇-hemitarate, and alkali addition salts thereof.

The skeletons of these compounds are active ingredients contained in licorice and are well-known compounds, but among these, glycyrrhetinic acid 3β-3-0-hemitarate disodium salt is
It is manufactured by the following method.

That is, it can be produced by using glycyrrhetinic acid (manufactured by Minophagen Pharmaceutical Co., Ltd., Ltd.) as a raw material, adding phthalic anhydride, and performing heating synthesis using a weak or strong base catalyst in an organic solvent. Here, pyridine, chloroform, methanol, ethanol, etc. can be used as the organic solvent, diethylamine, triethylamine, etc. can be used as the weak base catalyst, and 4-dimethylaminopyridine etc. can be used as the strong base catalyst.

In addition, glycyrrhetinic acid 3β-3-0-hemisuccinate is commercially available as the generic name rubenoxolone sodium.

While these compounds have a remarkable absorption promoting effect,
The side effects on biological mucous membranes are extremely small. Therefore, the compound of the present application can be used for transmucosal administration of poorly absorbed substances, for example, high molecular weight substances such as peptide hormones and interferons, which have conventionally had almost no pharmacological effect.
For example, nasal administration can provide sufficient absorption as in the case of conventional parenteral injections, and as a result, the desired therapeutic effect can be expected. In particular, great expectations are held regarding insulin as it will provide a preparation that is extremely useful for the treatment of diabetes.

When the compound of the present application is used as an absorption enhancer, it can be administered as an aqueous solution in which it is dissolved together with an applied amount of a poorly absorbed substance, and as a result of promoting transmucosal absorption, its network shape can be used as a nasal spray, It becomes possible to use it as a spray agent or the like.

The compound of the present invention can also be added to an applied amount of a poorly absorbable substance and applied topically or topically as an ointment using a conventional method.

In these cases, a sufficient effect of promoting transmucosal absorption can be expected by using the compound of the present invention in a content in the range of 0.1 to 10%.

Formulation Examples and Pharmacological Test Examples are listed below, but it goes without saying that the present invention is not limited to these Examples and Test Examples.

[Formulation Example K Manufacture of Nasal Drops Bovine insulin in a dosage unit per 0.1 ml of administration solution is dissolved in a 0.1 to 10% aqueous solution of disodium glycyrrhetinic acid 3β-3-0-hemitarate. Prepare.

Current insulin injection formulations come in two specifications: 40U and 100U, and when administering the same dose as the injection, the amount of solution administered through the nose is 0.4 to 1ml, which is sufficient for use as a nasal spray for humans. It is the amount of liquid.

In addition, isotonizing agents and other preservatives may be added as necessary.

Preparation of spray The solution for spray is prepared in the same manner as the nasal spray described above, and the container is filled with air or preon.

In this case, a container is used that can be administered in the form of a mist by manually pressing the container or compression device. The amount of spray is
Each dose is 0.1-0.2 ml, and 2-5 sprays are sufficient to administer a therapeutic amount of insulin.

[Pharmacological test example - Absorption promoting effect (1) Preparation of insulin administration solution Bovine insulin was prepared at a pH of 7.4 and 0.5%. OIM phosphate buffer ■ Glycyrrhetinic acid 3β-3-0 hemipthalate disodium at 0.
Insulin solutions of 100 U/ml were prepared by dissolving the insulin in the same buffer containing 1% am-so-hemiphthalate disodium glycyrrhetinic acid.

(2) Test animals: 3 to 6 Wistar rats (weighing around 250 g) per group.
The animals were placed in a dorsal position under bendoparbital anesthesia, and a cannula was inserted into the trachea to ensure breathing. A cannula was also inserted into the esophagus to prevent leakage of the administered solution.

(3) In the blood ei measurement experiment example, insulin was 2.5 and 10
Ulo was administered in an amount of 17 kg per ml, blood was collected from a cannula inserted into the femoral artery, and the heparin-pretreated blood was centrifuged (3,000 rpm, 10 minutes) to obtain plasma. Insulin was measured using an enzyme immunoassay (Insulin B-Test Wako, manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.). Blood sugar levels were measured using the mutarotase-glucose oxidase method (Glufus C-Test Wahoo, manufactured by Wako Tsumugi Pharmaceutical Co., Ltd.).

(4) Calculation method In addition, the absorption rate of insulin and the rate of decrease in blood sugar level were calculated using the following formula.

■Insulin absorption rate (%) Nasal: 0 of insulin during nasal administration of insulin
Area under the blood concentration time curve up to 4 hours (h・μU/mZ) Funtrol: O of insulin when insulin is not administered
Area under the blood concentration time curve up to 4 hours (h・μU/m) Intravenous injection: 0 of insulin during intravenous insulin injection
Area under the blood concentration time curve until the 4th hour (h・μU/mri) ■Decrease rate of blood sugar level (0%) m@gminority<0%)=! -Nasal (or intravenous) X1o.

Control nasal: Blood sugar level of 0 to 4 during nasal administration of insulin
Area under the blood concentration time curve (h - mg/di > Control: Area under the blood concentration time curve from 0 to 4 hours of blood glucose level when insulin is not administered (h - mg/d l) , ) Intravenous injection: Blood sugar level θ~4 during intravenous insulin injection
Area under the blood concentration time curve (h - mg/dl, ) (5) Results Figure 1 shows the results for non-insulin administration (control), insulin administration alone (alone, 1007 kg), and 1% glycyrrhetinic acid. Administration of insulin solution containing 3β-3-0-hemitarate disodium (hereinafter referred to as GAHP) (2,5
and 1007 kg), respectively.

Even when insulin was administered alone, the plasma insulin concentration increased significantly (p<0.01) 15 minutes after administration compared to Funtrol, and the time to reach the maximum blood concentration (t max) was 1 hour. Furthermore, the area under the blood concentration time curve (AUC6-4) up to 4 hours after administration was about 6 times that of Funtrol, but it did not lead to a decrease in blood sugar levels.

When adding 1% GARP, the change in blood insulin concentration with respect to the insulin dose shows that significant absorption was observed at 10 U/kg administration, but at 207 kg and 5 U/kg.
When administered, an increase in absorption was observed compared to when administered alone, but no significant difference was observed. Regarding the decrease in blood sugar level, a significant difference was observed between each blood collection time for any administration compared to the amount of Funtrol. FIG. 2 shows the time course of the blood sugar level at this time. No significant difference was observed between the control and insulin solution alone, but 1% GA) IF-containing insulin solution was added to
When 00/kg was administered, the blood sugar level was 45 minutes after administration.
mg, decreased to 7 dl, and 1 hour after administration, 22
It decreased to mg/dj2. Furthermore, the blood sugar level reducing effect of insulin was maintained until the end of the experiment.

On the other hand, at 207 kg and 5 υ/kg administration, 10 U
Although the degree of decrease in blood sugar level was smaller than that when administered at a dose of 100 mg/kg, it was significantly decreased compared to the control and when administered alone.

In addition, Table 1 below shows GA)iP, sodium glycyrrhetinate (hereinafter referred to as GA), and glycyrrhetinic acid 3β-
The results for 3-0-hemisuccinate (hereinafter referred to as GAH5) were compared with intravenous injection and subcutaneous injection.

It is the standard error above the mean of 3 to 6 rats.

In the case of no insulin administration, the AUC-a of insulin up to 4 hours is 28.7 fr-μU/ml, but when insulin is administered alone, it is 172 h·μU/ml.

In this way, insulin is absorbed even when given alone, but
The rate of decrease in blood sugar level (D%>) was 3.9%, and the pharmacological effect could not be fully demonstrated.

In contrast, 10 U of insulin containing 1% GAHP
/kg administration, the AUG of insulin, -4 is 1571
h・μU/m 1 is about 55 times the Funtrol value,
It was also 1.6 times that of GAI5. The blood sugar reduction rate at this time was 81.6%, which was almost the same value as that of intravenous injection.

In addition, the absorption rate of insulin with GAHP was significantly improved to 21.7% compared to 2.02% when insulin was used alone, and it had the best absorption promoting effect among the glycyrrhizin and glycyrrhetinate salts examined so far. It was something.

The intranasal administration of insulin in this test example is 2 to 10 times the amount in human terms, but judging from the blood sugar reducing effect described above, sufficient effects can be expected even with lower doses than these.

Acute toxicity test In the case of oral administration, LDsa is GAI (P is 1320
mg/kg.

GAI5 is 520 mg, 7 kg, and has low toxicity.

[Brief explanation of the drawing]

Figure 1 shows the plasma insulin concentration time curves after intranasal administration of insulin solutions containing 1% GAHP (2, 5 and 10 U/kg) to rats; , 5 and 10 U/kg) in rats after intranasal administration. Patent applicant Minophagen Pharmaceutical Co., Ltd. 2rA Hours, h Figure 1'' Hours, h

Claims (4)

[Claims] (1) General formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [However, in the formula, R is H-, HOOCCH_2CH_2C
Represents O- or ▲There are mathematical formulas, chemical formulas, tables, etc.▼, respectively. ] A transmucosal absorption enhancer for poorly absorbable substances containing glycyrrhetinic acid and its derivatives as shown in the following, and pharmaceutically acceptable salts thereof as active ingredients. (2) General formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [However, in the formula, R is H-, HOOCCH_2CH_2C
Represents O- or ▲There are mathematical formulas, chemical formulas, tables, etc.▼, respectively. ] A peptide hormone transmucosal absorption enhancer containing glycyrrhetinic acid and its derivatives as shown in the following, and their pharmaceutically acceptable salts as active ingredients. (3) General formulas for poorly absorbable substances: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [However, in the formula, R is H-, HOOCCH_2CH_2C
Represents O- or ▲There are mathematical formulas, chemical formulas, tables, etc.▼, respectively. ] A nasal spray characterized by adding glycyrrhetinic acid and its derivatives represented by these, and their pharmaceutically acceptable salts. (4) The nasal spray according to claim 3, wherein the poorly absorbed substance is a peptide hormone.