JPH0228121A - Transmucosal absorbefacient and pernasal administering agent using the same absorbefacient - Google Patents

Abstract

PURPOSE:To obtain a transmucosal absorbefacient, containing a specific glycyrrhetinic acid derivative as an active ingredient and having remarkable transmucosal absorbefacient action on hardly absorbable medicines, such as peptide hormone, with extremely minute side effects on mucous membranes in living bodies. CONSTITUTION:A transmucosal absorbefacient, containing a compound expressed by the formula [X is residue of olean-11,13(18)-dine-3beta,30-diol 3beta,30-di-O- hemiphthalate, 18beta-olean-12-ene-3beta,30-diol 3beta,30-di-O-hemiphthlate or 18beta- olean-9(11),12-diene-3beta-30-diol 3beta,30-di-O-hemiphthalate reside] and a pharmaceutically acceptable salt thereof as an active ingredient, promoting transmucosal absorption of the afore-mentioned medicines, e.g., insulin (remedy for diabetes), hardly absorbed from nasal mucous membranes due to a high molecular weight and administered merely by injection and enabling transmucosal, especially pernasal administration.

Description

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

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

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, nasal sprays and suppositories are being researched as preparations that avoid these situations and enable home administration, but insulin, for example, has a molecular weight of about 6,000, Therefore, it is extremely difficult to absorb from the nasal mucosa or rectal mucosa.

Therefore, in recent years, substances that can promote the absorption of insulin and other substances have been investigated, and substances with surface-active effects such as bile acids, medium-chain fatty acids, nonionic or ionic synthetic surfactants, and saponins have been found to have an absorption-promoting effect. Something has become known.

The inventors of the present application have discovered the active ingredients of licorice, glycyrrhizin and glycyrrhetinic acid, which have surfactant action and are used not only as internal medicines but also as external medicines such as eye drops and ointments, and are added to cosmetics and foods as sweeteners. It has been discovered that the derivatives glycyrrhetinic acid 3β-O-hemisuccinate and its salts and glycyrrhetinic acid 3β-O-hemisuccinate and its salts have an effect of promoting transmucosal absorption of insulin, glucagon, etc.

Therefore, the inventors of the present invention developed a new derivative of glycyrrhetinic acid, oleane-11.13(18)-diene-
3β, 30-diol 3β, 30-di0-hemiphthalate, 18β-ocyan-12-ene-3β, 30-diol 3β, 30-di0-hemiphthalate, 18β-
Olean-9(H), tz-diene-3β, 30-diol 3β, 30-di-0-hemitarate, and their alkali addition salts were investigated for promoting transmucosal absorption of insulin.

As a result of various studies, the inventors of the present application have discovered that the above-mentioned compound has 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 something.

Compounds useful as absorption enhancers in the present invention have the general formula (1): where X is oleane-11.13(18)-diene-
3β, 30-diol 3β, 30-diO-hemitarate, 18β-ocyan-12-ene-3β, 30-diol 3β, 30-diO-hemitarate, or 18
β-diene-9 (11), 12-diene-3β,
It is a residue of 30-diol 3β, 30-diO-hemitarate] and a pharmaceutically acceptable salt thereof.

The compound included in the above general formula (1) is represented by the formula: Formula: [However, in the formula, R and R1 are C;
β.

30-diol 3β, 30-di○-hemitarate (
Hereinafter abbreviated as HetAD DHPh) and its alkali addition salt, 18β-odoan-12-ene-3β,3Q
- diol 3β, 30-di-0-hemitarate (hereinafter abbreviated as DG-DHPh) and its alkali addition salt,
and the formula: [wherein Il and R1 are 18β-orian-12-ene-3β, 30
-3β,30-diol 3β,30-di-O-hemiphthalate (hereinafter abbreviated as HomAD-DHPh) and its alkali addition salt.

These compounds can be obtained by the method described in JP-A-60-260540. That is, Olean-11.13
(18)-diene-3β, 30-diol, 18β-orian-12-ene-3β, 30-diol or 18β
-Olean-9(11), 12-diene-3β, can be synthesized by adding phthalic anhydride to 30-diol and 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, tonoethylamine, etc. can be used as the weak base catalyst, and 4-dimethylamine, nobiridine, etc. can be used as the strong base catalyst.

While these compounds have a remarkable absorption promoting effect,
The side effects on biological mucous membranes are extremely small. Therefore,
When administering difficult-to-absorb substances such as peptide hormones and high-molecular-weight substances such as interferon, the present compound can be administered by transmucosal administration (for example, intranasal administration), which previously had no pharmacological effects, or by conventional injections. Similarly, sufficient absorbability can be obtained, and as a result, desired therapeutic effects can be expected. In particular, there are great expectations 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 appropriate amount of a poorly absorbed substance, and membrane absorption is promoted, so that the dosage form is a nasal spray. , a spray, etc.

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

In these cases, the present compound can be expected to have a sufficient effect of promoting transmucosal absorption by using the compound in a content of Q, (11 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: Manufacture of nasal spray HetAD-DHPh-2Na 5alt, DG
-DHPh -2Na 5alt and HomAD -
It is prepared by dissolving maggot insulin in a dosage unit per 0.1 ml of the administration solution in a 0.01-10% aqueous solution of DHPh-2Na 5alt.

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, tonicity agents, stabilizers, preservatives, etc., which are acceptable for formulation, are added as necessary.

Manufacture of spray solution The solution for spray is prepared in the same manner as the nasal spray mentioned above, and if the container is filled with air or Freon, it is administered as a mist by manually pressing the container or compression device. Use containers that can be used. Spray amount is 0.1-0.2m at a time
1, and 2 to 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 ■HetAD
-DHPh -2Na 5alt containing 0.25% HomAD -DHPh -2Na 5alt
The same buffer containing 1% DG-DHPh-2Na 5
Alt was dissolved in the same buffer containing 0.01-1%, and 1% of each
00 U/ml insulin solutions were prepared.

(2) Test animals: 3 to 6 Wistar rats (weighing around 250 g) per group.
The animals were placed in a dorsal position under ventobarbital 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) Measurement of blood concentration In this experimental example, the amount of insulin was 10 U10.1 ml.
/kg, blood was collected from a cannula inserted into the femoral artery, and the heparin-pretreated blood was centrifuged (
3,000 rpm for 10 minutes) to obtain plasma. Insulin was measured using an enzyme immunoassay (Insulin B-Test Wafu, Wako Tsumugi IM). Blood sugar levels were measured using the mutarotase-glutose oxidase method (Glufus C-Test Wako, 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.

■Relative absorption rate of insulin (%) Nasal: Bottom line of the blood concentration time curve from 0 to 4 hours when insulin is administered nasally (h・μU/ml) Control: Insulin concentration when insulin is not administered O~
Area under the blood concentration time curve up to the 4th hour (h・μU/ml) Intravenous injection: Insulin θ during intravenous insulin injection
Bottom line of blood concentration time curve up to 4 hours (h・μU/ml) ■Decrease rate of blood sugar level (DX> Nasal: 0 to 4 of blood sugar level during nasal administration of insulin)
Lower end of the blood concentration time curve up to the 4th hour (h - mg/dl) Control: Lower area of the blood concentration time curve from 0 to 4th hour of the blood glucose level when insulin is not administered (h - mg/dl) Intravenous injection: Blood sugar level during intravenous insulin injection 00-4
The lower end of the blood concentration time curve (h - mg/dl) (5> Results Figure 1 shows the results of insulin non-administration (control), insulin alone administration (!Ji-Germany, to U/kg), 0 ．．
2 shows the blood concentration time curves of each insulin after administration of an insulin solution containing 01-1% DG-DHPh-2Na5alt (10 U/kg).

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

DG-DHP versus dose of insulin IOU/kg
Regarding changes in the blood concentration of insulin when h-2Na 5alt was added at 1 degree, remarkable absorption was observed when 0.5% was added, and a maximum blood concentration of 4.75 mU/ml was obtained 30 minutes after administration.

Also, 0.1-1% DG-DHPh-2Na 5al
At an addition concentration of t, t maw is 30 minutes and 0.05
%, t and max extended to 1 hour, and at 0.01%, the blood insulin concentration increased even at the 4th hour at the end of the experiment.

Therefore, the insulin absorption behavior of the compound of the present invention differs depending on the added concentration, and when the concentration is high, a significant increase in blood fIR is obtained within a short period of time, which is the property of conventional absorption enhancers, and the concentration increases. When the concentration was low, a sustained blood concentration was obtained.

FIG. 2 shows the time course of the blood sugar level at this time. No significant difference was observed between control and single administration, but 0.05, 0.5 and 1% DG-DHPh-2
In the Na 5Alt addition group, 30 mg/d 1 hour after administration.
1 hour after administration in the 0.1% addition group.
It decreased to mg/dl. Although no decrease in blood sugar level was observed in the -10.01% addition group, it decreased over time compared to the control and single administration group.

Furthermore, Table 1 below compares the effect of the glycyrrhetin derivative according to the present invention on the nasal absorption rate and blood glucose reduction rate of insulin after intranasal administration of insulin to rats, compared with intravenous injection and subcutaneous injection. .

Table 1 f of insulin after rat intraascending administration of insulin! Effect control of glycyrrhetinic acid TS conductor on nasal absorption force and blood glucose reduction rate Nasal alone DC-DIIPh-2N engineering 5alt 11o-^D-DHPh-28 wealth s+altflat
＾Ko DHPh-2Nm 5alt Intravenous Subcutaneous Enhancer Concentration % Dose ＾UC,.

Ship to U/ h・μU/m 28.7±5.1 +72±14.2 2059*447 1543±197 7092±1416 5181±966 568f17.8 3576±507 1456±260 blood sugar Decrease rate % 3.9±11,0 12.0 71.3±6.5 60.1±42 73.6±4.6 75.8±3.3 61.8±2.0 67.1±4.1 81.8±1,0 76.2±4.4 Each value is the mean main standard error of 3 to 6 rats.

Relative absorption rate % 2.02 10.71 28.61 21.34 99, 55 72, 133 97, 16 40.2 In the case of no insulin administration, the AUC of insulin up to the 4th hour was 28.7 h・μU/ml, but when insulin is administered alone, it becomes 172 h・μU/ml.

On the other hand, when administering 10 U/kg of insulin containing 0.5% DG-DHPh-2Na 5alt,
AUCo-4 of insulin is 7092h・μIJ/ml
It was about 247 times the control value, and the blood sugar reduction rate at this time was 74%.

This is 99, assuming the absorption rate of intravenous injection to be 100%.
6X, and no significant difference was observed between the two in blood sugar reduction rate. Also, o, oi%DG-DHPh-2
With the addition of Na 5alt, the absorption rate was 10.7% and the blood sugar reduction rate was 12%, indicating a slight pharmacological effect.

As mentioned above, DG -DHPh -2Na 5alt
Of the glycyrrhetinic acid derivatives examined so far, it has the best absorption promoting effect, and even when added at a low concentration, it sufficiently exerted its absorption promoting effect.

In addition, 1 HomAD-DHPh-2Na 5alt
% added insulin 100/Kg dose, the AUCo -4 of insulin was 0 at 6922 h-μtJ/ml.
．． 5% DG-DHPh-2Na 5alt group, however, DGDHPh-28@5alt is the average value of 2 animals.

It showed almost the same level of absorption. On the other hand, 0.25% HetAD
-DHPh -2Na The absorption rate was about 7% in the 5alt group.

If the nasal administration of insulin in this test example is compared to humans, it will be 10 times the amount, but the absorption rate is comparable to intravenous injection and 2.5 times that of subcutaneous injection, which is the usual administration method. from,
Adequate effects are expected even when administered at a low dose equal to or lower than that of commercially available injections.

Acute toxicity test Acute toxicity test (L
The results of measuring D6゜〉 are as follows, indicating low toxicity.

Animal used: ICR mouse, 7 weeks old, male, weight 33-3
5gLDao calculation: Van der Waerden
Dimensions: DG-DHPh-2Na 5alt: 1257
~1520mg/kgHetAD-DHPh-2Na
5alt: 628-793mg/kg1 (omAD
-DHPh-2Na 5alt: 1283-149
9mg/kg

[Brief explanation of the drawing]

Figure 1 shows various concentrations of DG -DHPh -2Na 5
Figure 2 shows the plasma insulin concentration time curve after intranasal administration of alt-containing insulin solution (10 U/kg) to rats.
FIG. 2 is a diagram showing changes in blood glucose levels after intranasal administration of an lt-containing insulin solution (10 U/kg) to rats.

Claims (4)

[Claims] (1) General formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, X is olean-11,13(18)-diene-
3β,30-diol 3β,30-di-O-hemiphthalate, 18β-olean-12-ene-3β,30-diol 3β,30-di-O-hemiphthalate, or 18
β-oleane-9(11),12-diene-3β,30
- a residue of diol 3β,30-di-O-hemiphthalate] A transmucosal absorption enhancer for a poorly absorbable substance containing a glycyrrhetinic acid derivative represented by the following and a pharmaceutically acceptable salt thereof as active ingredients. (2) General formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, X is olean-11,13(18)-diene-
3β,30-diol 3β,30-di-O-hemiphthalate, 18β-olean-12-ene-3β,30-diol 3β,30-di-O-hemiphthalate, or 18
β-oleane-9(11),12-diene-3β,30
- a residue of diol 3β,30-di-O-hemiphthalate] A peptide hormone transmucosal absorption enhancer containing a glycyrrhetinic acid derivative represented by the following and a pharmaceutically acceptable salt thereof as active ingredients. (3) General formulas for poorly absorbable substances: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, X is olean-11,13(18)-diene-
3β,30-diol 3β,30-di-O-hemiphthalate, 18β-olean-12-ene-3β,30-diol 3β,30-di-O-hemiphthalate, or 18
β-oleane-9(11),12-diene-3β,30
- a residue of diol 3β,30-di-O-hemiphthalate], and a pharmaceutically acceptable salt thereof. (4) The nasal preparation according to claim 3, wherein the poorly absorbed substance is a peptide hormone.