JP2720899B2 - Transmucosal absorption enhancer and nasal administration agent using the same - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a glycyrrhetinic acid derivative, a transmucosal absorption enhancer of a poorly absorbable substance containing a medically acceptable salt thereof as an active ingredient, and the like.

[Prior art and problems] Among the conventionally used pharmaceuticals, poorly absorbable substances such as peptide hormones, for example, insulin, have been administered only by injection to diabetic patients and the like.

However, injection methods of these poorly-absorbable substances, such as insulin, have the drawbacks that they give pain at the time of injection, and that tissue is thickened at the injection site by continuous administration for a long period of time.

On the other hand, nasal drops and suppositories have been studied as preparations that avoid these situations and allow home administration.For example, insulin has a molecular weight of about 6,000, It is extremely difficult to be absorbed from mucous membrane or rectal mucosa.

Therefore, in recent years, substances that promote absorption of insulin and the like have been studied, and substances having a surfactant effect such as bile acids, medium-chain fatty acids, nonionic or ionic synthetic surfactants, and saponins have an absorption promoting effect. It has become known that there is.

The present inventors have a surfactant activity, glycyrrhizin, glycyrrhetinic acid and glycyrrhetinic acid as active ingredients of licorice active ingredients, which are used as topical drugs for ophthalmic ointments and ointments in addition to oral medicines, and are further added to foods as cosmetics and sweeteners. It has been discovered that the derivatives glycyrrhetinic acid 3β-O-hemisuccinate and salts thereof and glycyrrhetinic acid 3β-O-hemiphthalate and salts thereof have a transmucosal absorption promoting effect on insulin and glucagon.

Therefore, the present inventors have newly developed a derivative of glycyrrhetinic acid as 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, 18β-olean-9 (11), 12
-Diene-3β, 30-diol 3β, 30-di-O-hemiphthalate 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 found that the above compounds exert a remarkable effect on the promotion of transmucosal absorption of high molecular weight peptide hormones including insulin and glucagon, and have completed the present invention. It is.

In the present invention, a compound useful as a compound which promotes absorption is represented by the general formula (1): Wherein 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 18β-olean-9 (11), 1
2-diene-3β, 30-diol 3β, 30-di-O-hemiphthalate], and pharmaceutically acceptable salts thereof.

The compound included in the general formula (1) has a formula: Olean-11,13 (18) -diene-3β, 30- represented by
Diol 3β, 30-di-O-hemiphthalate (hereinafter referred to as Het
AD-DHPh) and its alkali addition salts, 18β-olean-12-ene-3β, 30-diol 3β, 30-di-O-hemiphthalate (hereinafter referred to as DG-DHP)
h) and its alkali addition salts, and the formula: 18β-Olean-12-ene-3β, 30-3β,
30-diol 3β, 30-di-O-hemiphthalate (hereinafter abbreviated as HomAD-DHPh) and its alkali addition salts.

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-olean-12-ene-3β, 30-diol or 18β-olean-9 (11), 12.
Phthalic anhydride to diene-3β, 30-diol,
It can be synthesized in an organic solvent using a weak base catalyst or a strong base catalyst. Here, pyridine, chloroform, methanol, ethanol and the like are used as the organic solvent, and diethylamine, triethylamine and the like can be used as the weak base catalyst, and 4-dimethylaminopyridine and the like can be used as the strong base catalyst.

While these compounds have a remarkable absorption promoting action, the side effects on living mucous membranes are extremely small. Therefore, poorly absorbable substances by the compound of the present application, for example, in the administration of high molecular weight substances such as peptide hormones and interferons, transmucosal administration, which had almost no pharmacological effect,
For example, even by nasal administration, sufficient absorbability can be obtained as in the case of conventional injections, and as a result, a predetermined therapeutic effect can be expected. In particular, insulin is greatly expected to provide an extremely useful preparation for treating diabetes.

When the compound of the present invention is used as an absorption enhancer, it can be administered as an aqueous solution dissolved with an applied amount of a poorly absorbable substance, and as a result, transmucosal absorption is promoted. It can be used as a spray or the like.

The compound of the present invention can also be added topically to a suppository or ointment by a conventional method after being added to an applied amount of the poorly absorbable substance.

In these cases, a sufficient transmucosal absorption promoting effect can be expected by using the compound of the present invention at a content in the range of 0.01 to 10%.

The 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 Examples] Production of nasal drops HetAD-DHPh-2Na salt, DG-DHPh-2Na salt and HomA
D-DHPh-2Na salt 0.01 to 10% aqueous solution, administration solution 0.1ml
It is prepared by dissolving per unit dose of bovine insulin.

The current insulin injection formulation is two standards of 40U and 100U, and when the same dose as the injection is administered, the amount of liquid administered in the nasal drop is 0.4 to 1 ml, which is a sufficient amount as a nasal drop for humans. is there.

In addition, if necessary, pharmaceutically acceptable isotonic agents, stabilizers and preservatives are added.

Preparation of Spray Preparation A solution for spraying is prepared in the same manner as in the above nasal drops, and the container is filled with air or freon.

In this case, a container or a container which can be administered as a mist by pressing the device for compression by hand or the like is used. The nebulization volume is 0.1-0.2 ml per dose, and two to five nebulizations can sufficiently administer a therapeutic amount of insulin.

[Pharmacological test example] Absorption-promoting action (1) Preparation of insulin administration solution HomAD-DHPh-2Na salt containing bovine insulin, pH 7.4, 0.01M phosphate buffer HetAD-DHPh-2Na salt containing 0.25% The same buffer containing 1% DG-DHPh-2Na salt was dissolved in the same buffer containing 0.01 to 1% to prepare an insulin solution of 100 U / ml each.

(2) Test animals Wistar rats (body weight: about 250 g) were grouped into groups of 3 to 6 rats under pentobarbital anesthesia, dorsal position was fixed, and respiration was ensured by inserting a cannula into the trachea. A cannula was inserted into the esophagus to prevent leakage of the administered solution.

(3) Measurement of blood concentration In this experimental example, insulin was administered at 10 U / 0.1 ml / kg, blood was collected from a cannula inserted into the femoral artery, and heparin-pretreated blood was centrifuged (3,000 rpm, 1
0 min) to obtain plasma. Insulin was measured using an enzyme immunoassay (insulin B-Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.). The blood glucose level was measured using the mutarotase / glucose oxidase method (glucose C-Test Wako, manufactured by Wako Pure Chemical Industries, Ltd.).

(4) Calculation method In addition, the absorption rate of insulin and the reduction rate of blood glucose level were calculated from the following equations.

Relative absorption rate of insulin (%) Nasal: Area under the blood concentration-time curve from 0 to 4 hours after nasal administration of insulin (h · μU / ml) Control: 0 to insulin without insulin administration
Area under the blood concentration time curve up to 4 hours (h · μU / ml) Intravenous injection: Area under the blood concentration time curve from 0 to 4 hours of insulin at the time of intravenous insulin injection (h · μU / ml) Blood sugar reduction rate (D%) Nasal: The area under the blood concentration time curve from 0 to 4 hours of the intranasal insulin administration (h · mg / dl) Control: The blood glucose level from 0 to 4 hours without the administration of insulin Area under the blood concentration-time curve (h · mg / dl) IV: Area under the blood concentration-time curve (h · mg / dl) from 0 to 4 hours of the blood glucose level during intravenous insulin injection (5) Results FIG. 1 shows insulin non-administration (control), insulin alone administration (alone, 10 U / kg), 0.01-1% DG-DHPh
FIG. 2 shows the blood concentration-time curves of the respective insulins after administration of an insulin solution containing −2Na salt (10 U / kg).

Even when insulin alone was administered, the plasma insulin concentration significantly (p <0.01) increased 15 minutes after administration compared to the control,
The time to reach the maximum blood concentration (t max) was 1 hour. In addition, the area under the blood concentration time curve up to 4 hours after administration (AU
C _0-4 ) was about 6 times that of the control, but did not reach the point where the blood glucose level was reduced.

DG-DHPh-2Nasa for a dose of 10 U / kg of insulin
The change in blood concentration of insulin at the lt addition concentration is 0.5
%, Marked absorption was observed at 30 minutes after administration.
A maximum blood concentration of 75 mU / ml was obtained.

Further, at a concentration of 0.1 to 1% DG-DHPh-2Na salt added, tmax is 30 minutes, at 0.05%, tmax is extended to 1 hour, and at 0.01%, insulin is increased even at the 4th hour at the end of the experiment. In the blood increased. Therefore, the compound of the present invention has a difference in insulin absorption behavior depending on the added concentration, and when the concentration is high, a marked increase in blood concentration within a short time, which is a property of a conventional absorption enhancer, is obtained, and the concentration is increased. When it was low, a sustained blood concentration was obtained.

FIG. 2 shows a time transition of the blood sugar level at this time.
No significant difference was observed between the control and the single administration, but decreased to 30 mg / dl 1 hour after administration in the 0.05, 0.5 and 1% DG-GHPh-2Na salt added groups, and 1 hour after administration in the 0.1% added group. Reduced to 50mg / dl. On the other hand, no decrease in blood glucose level was observed in the 0.01% addition group, but it decreased over time as compared with the control or the single administration group.

In addition, Table 1 below shows the ratio of insulin to nasal absorption and blood glucose reduction after intranasal administration of insulin to rats.
FIG. 4 shows the effects of the glycyrrhetin derivative according to the present invention compared with intravenous injection and subcutaneous injection.

In the case of non-administration of insulin, the AUC _0-4 of insulin up to the fourth hour is 28.7 h · U / ml, whereas that of insulin alone is 172 h · U / ml.

In contrast, in the case of administration of 10 U / kg of insulin to which 0.5% of DG-DHPh-2Na salt was added, AUC _0-4 of insulin was 7092 h.
-With μU / ml, the control value was about 247 times, and the blood glucose reduction rate at this time was 74%. This increases the rate of intravenous absorption
When it was set to 100%, it was equivalent to 99.6%, and there was no significant difference between the two in the blood glucose reduction rate. In addition, 0.01% DG
With the addition of -DHPh-2Na salt, the absorption rate was 10.7% and the blood glucose reduction rate was 12%, indicating a slight but pharmacological effect.

As described above, DG-DHPh-2Na salt has the best absorption promoting effect among the glycyrrhetinic acid derivatives studied so far, and exhibited a sufficient absorption promoting effect even at a low concentration.

In addition, with administration of 10 U / Kg of insulin to which HomAD-DHPh-2Na salt was added at 1%, AUC _0-4 of insulin was 6922 h · μU.
/ ml showed almost the same absorption as the 0.5% DG-DHPh-2Na salt group. On the other hand, the absorption rate was about 7% in the group of 0.25% HetAD-DHPh-2Na salt.

When the nasal dose of insulin in this test example is converted to human, it is 10 times the amount.However, the absorption rate is comparable to that of intravenous injection, and is 2.5 times that of subcutaneous injection, which is the usual administration method. Sufficient effects are expected even with administration of a low dose equal to or less than that of an injection.

Acute toxicity testing the claimed compounds, the acute toxicity test by oral administration (LD ₅₀₎ result of measuring are as follows: low toxicity.

Animals used: ICR strain mice 7-week-old male body weight 33~35g LD ₅₀ calculated: Van der Waerden's method DG-DHPh-2Na salt: 1257~1520mg / kg HetAD-DHPh-2Na salt: 628~793mg / kg HomAD-DHPh −2Na salt: 1283-1499mg / kg

[Brief description of the drawings]

FIG. 1 is a diagram showing plasma insulin concentration-time curves after intranasal administration of various concentrations of DG-DHPh-2Na salt-containing insulin solution (10 U / kg) to rats, and FIG. 2 is a diagram showing various concentrations of DG-DH.
It is a figure which shows the change of the blood glucose level after the intranasal administration of the insulin solution containing Ph-2Na salt (10U / kg) to a rat.

Claims (4)

(57) [Claims] (1) a general formula: Wherein 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 18β-olean-9 (11), 1
2-diene-3β, 30-diol 3β, 30-di-O-hemiphthalate], a glycyrrhetinic acid derivative represented by the formula: Transmucosal absorption enhancer. 2. The general formula: Wherein 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 18β-olean-9 (11), 1
Transgenic mucosa of a peptide hormone containing a glycyrrhetinic acid derivative represented by the formula (1): Absorption promoter. 3. The poorly-absorbable substance has a general formula: Wherein 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 18β-olean-9 (11), 1
Which is a residue of 2-diene-3β, 30-diol 3β, 30-di-O-hemiphthalate], and a pharmaceutically acceptable salt thereof. Dosing agent. 4. The composition for nasal administration according to claim 3, wherein said poorly-absorbable substance is a peptide hormone.