JPH02306919A - Nasal mucosal vascular penetrative inhibitor - Google Patents

Abstract

PURPOSE:To obtain a nasal mucosal vascular penetrative inhibitor, containing an N-acetylneuraminate as an active ingredient, capable of inhibiting vascular penetrativity accelerated by antigen antibody reaction and useful for rhinitis and allergic rhinitis, etc., such as improvement in swelling of nasal mucosa and hypersecretion of pituita into the nasal cavity, etc. CONSTITUTION:An inhibitor containing a compound expressed by the formula (Z is lithium, potassium, sodium, etc., when n is 1 and calcium barium or magnesium when n is 2) as an active ingredient. The compound expressed by the formula can be administered in a local administration form, e.g. converted into fine particles and sprayed in the nasal cavity, dissolved in a solution and dripped into the nasal cavity or contained in a disk and applied into the nasal cavity. The dose thereof is 0.1-50mg as the compound expressed by the formula, 0.02-0.5ml at a time in the case of an aqueous solution for spraying and 0.01-0.05ml per drop in the case of dripping or 5-50mg in the case of powdery spray.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention relates to a nasal mucosal vascular permeability inhibitor, particularly for suppressing vascular permeability in living organisms, and particularly for suppressing nasal obstruction due to swelling of the nasal mucosa and secretion of nasal secretions into the nasal cavity. The present invention relates to a vascular permeability inhibitor containing N-acetylneuraminate that has the effect of improving symptoms.

[Prior Art and Problems to be Solved] Rhinitis and allergic rhinitis are among diseases in the field of otorhinolaryngology. Rhinitis appears during climate changes and colds, and is triggered by chemical, microbiological or mechanical stimuli. Symptoms include rhinorrhea, nasal obstruction, and obstructive nasal voice.

In addition, allergic rhinitis is a nasal disease resulting from an allergic reaction in the nasal mucosa. Symptoms include paroxysmal and recurrent sneezing, watery nasal discharge, and nasal obstruction.The mechanism of occurrence of these rhinitis symptoms is that the above irritation causes hyperemia and congestion in the nasal mucosa, which increases vascular permeability. , swelling of the nasal mucosa, hypersecretion of nasal secretions, and finally symptoms of nasal obstruction.

Conventionally, the mechanism of action of the drugs for rhinitis and allergic rhinitis mentioned above is to stimulate α-receptors and excite sympathetic nerves, thereby constricting peripheral blood vessels, thereby causing hyperemia, congestion, and swelling of the nasal mucosa. was to be removed.

However, conventional drugs have problems such as the risk of increased blood pressure due to vasoconstriction and short-lasting drug effects. Furthermore, corticosteroids have also been used for their anti-inflammatory mechanism, but they have had safety issues.

In recent years, drugs developed for the treatment of allergic rhinitis have the effect of inhibiting the release of various chemical mediators.

By the way, various studies have been conducted on the effective pharmacology of N-acetylneuraminic acid, and it is currently reported that it has anti-inflammatory effects [Robert El Hirshiya, The Journal of Immunology -Volume 127, No. 5,
Pages 1740-1743, 1981; B. Goolug et al. Agents and Actions Vol. 8, No. 5, 543-
545 pages 1978; Hiromi Ito et al. Pharmacology and Treatment 13
Vol. 7, pp. 479-494, 1985] N-acetylneuraminic acid is neutralized with an alkali metal or alkaline earth metal hydroxide or a carbonate thereof, and then treated with a corresponding alkali metal salt or alkaline earth metal hydroxide, respectively. It can be obtained by separating the similar metal salt from the system (JP-A-62-T223123).

The applicant previously filed an application for an expectorant and cell tissue repair agent containing a salt of N-acetylneuraminic acid as an active ingredient (Japanese Patent Application Laid-open Nos. 61"-289037, 61-68418, and 64-3193). .

However, no drug having the effect of suppressing vascular permeability in the nasal mucosa containing N-acetylneuraminate as a main component has been known so far.

[Purpose of the invention]

An object of the present invention is to provide a novel nasal mucosal vascular permeability inhibitor containing N-acetylneuraminic acid salt as a main component.

[Means to solve the problem]

The present inventors have found that by using N-acetylneuraminate, vascular permeability, which is increased by antigen-antibody reactions, is suppressed, thereby improving swelling of the nasal mucosa and hypersecretion of nasal secretion into the nasal cavity. The inventors have discovered that nasal obstruction symptoms caused by accumulation of nasal secretions in the nasal cavity, difficulty in draining nasal secretions, etc. can be improved, and the present invention has been achieved.

That is, the present invention relates to a nasal mucosal vascular permeability inhibitor containing a compound represented by the general formula: as an active ingredient.

(However, when n=1, Z is lithium, potassium, sodium, ammonium or organic ammonium, and n
When is 2, Z represents calcium, barium or magnesium. ) In the present invention, the compound of the present invention can be administered in a topical administration form, for example, micronized and sprayed into the nasal cavity, dissolved in a solution and instilled into the body cavity, contained in a disk and applied to the nasal cavity, etc.

Hereinafter, the effects and formulation of the compound of the present invention will be explained in more detail with reference to Examples and Reference Examples.

Example 1 (1) Experimental Animals Male Wistar rats weighing 2OL-220g were used.

(2) Experimental material antiserum is rat anti-ovalbumin 1gε antiserum (antibody titer =
1/32 to 1/64), and ovalbumin was used as the antigen. The test agents were sodium salt of N-acetylneuraminic acid (manufactured by Mect) and DSCG (Sodium c
romoglycate) (manufactured by Fujisawa Pharmaceutical Co., Ltd.) was used.

(3) Experimental method Anti-ovalbumin IgB serum 0.5 mA! / 100
g was injected into the tail vein of rats to passively sensitize them, and experiments were conducted 24 hours later. Bentoparbital sodium (3Q
The animals were anesthetized by intraperitoneal administration of tng/kg) and fixed in the dorsal position. To maintain breathing, the trachea was incised, a cannula was inserted, and the esophagus was ligated.

Insert a cannula for nasal perfusion through the tracheostomy, connect it to a perfusion pump, and add 0.25% of physiological saline heated to 37°C.
Perfuse at a flow rate of m1/min, and remove the fluid flowing out from the body by 1
Sampled for 0 minutes. Next, 4% Blue IJ Ant Blue (5 mg/kg) was administered through the tail vein and collected for 10 minutes in the same manner. Perfuse antigen solution (ovalbumin) for 10 minutes,
Caused an allergic reaction. Irrigate with saline again,
Perfusate was collected every 10 minutes. The samples collected were 300
After centrifugation at 0 rpm for 10 minutes, the brilliant blue absorbance of the supernatant was measured at 620 nm.

In addition, as shown below, the test group was the following three groups, and the test drug was mixed in the perfusate and administered to rats in groups 1 to 2 before intravenous injection of the dye. The effects of this experiment were investigated.There were 10 rats in each group.

Group 1 2N-acetylneuraminic acid sodium salt 100μ
Administer g/ml.

■Group: DSCo 100 μg/mj! administered.

Group ■: Physiological saline administration (4) Results Figure 1 (5) Judgment From Figure 1, it can be seen that the sodium salt of N-acetylneuraminic acid has a stronger inhibitory effect on nasal mucosal vascular permeability than DSCG. I understand.

Example 2 Acute toxicity test Oral, subcutaneous, and intraperitoneal injection of N-acetylneuraminic acid sodium salt to mice, rats, and guinea pigs.
Acute toxicity tests by intravenous injection and inhalation were conducted as follows.

(1) Test animals: ICR mouse, 6 weeks old, SD rat, 6 weeks old, Hartley ()Iar
tley) strain guinea pig, 6 weeks old (2) Drug concentration 20% (W/V) Dissolved in distilled water (3) Number of Lebel animals: 10 (4) Observation period 14 days (5) L D s. Table 2 shows the results of the Probit calculation method.

Table 2 Acute toxicity test of N-acetylneuraminic acid sodium salt For inhalation, the compound of the present invention was exposed in the form of a mist for 1 hour.

95% confidence limits are in parentheses.

Example 3 Simple acute toxicity test A simple acute toxicity test by intravenous injection of the test agent into mice was conducted as follows.

1. Test materials and methods (1) Test agent lithium salt, potassium salt of N-acetylneuraminic acid,
Barium salt and magnesium salt (manufactured by MECT@) were used.

(2) Test animals DDL male mice Body weight at start of test: 17.7-21.1 g Number of Rubel animals = 3 (3) Room temperature: 23 ± 1°C Humidity: 55 ± 7% (4)
Administration route: Intravenous (5) Administration method and dosage Dissolve the above test agent in physiological saline, adjust the concentration so that the amount of solution administered is 0.2 ml per 20 g of mouse body weight, and inject into the tail vein. did. Dosage is 500.1000
and 3 doses of 2000 mg/kg.

(6) Observation of general symptoms and mortality status General symptoms and mortality were observed from immediately after administration until 7 days after administration.

2. Results (1) Mortality rate The mortality rate is shown in Table 3.

At 1000 and 2000 mg/kg, 3 out of 3 patients died with the magnesium salt, barium salt, and potassium salt of N-acetylneuraminic acid.

At 500 mg/kg, 3 out of 3 patients died with N-acetylneuraminic acid barium salt. There were no other deaths.

(2) General symptoms Death cases were accompanied by clonic convulsions and urinary incontinence, and most died within 1 minute immediately after administration. A small number of surviving cases showed suppression of locomotor activity, but recovered within 1 hour.

Next, we will discuss the dosage form of the nasal spray of the present invention.Administration method> The dosage may be adjusted as appropriate depending on the symptoms of the patient, but the recommended dose for one administration is 0.1 to 50 o+g of the compound of the present invention, and 3 to 5 g of this compound.
Spray 1 to 3 times or drop 2 to 4 drops into the nasal cavity every hour.

In the case of an aqueous solution, the spray amount is 0.02 to 0.5 ml per drop. O1 to 0.05 m1'. Moreover, in the case of a powder spray, it is 5-50a+g.

Note that the number of administrations per day is not limited based on the results of pharmacokinetic tests and acute toxicity tests.

Preparation> In the case of an aqueous solution, the proportion of the compound of the present invention in the preparation is 0.
5 to 10.0% by weight is suitable. N-acetylneuraminic acid sodium salt is most stable in the pH range of 5.5 to 6.0 and is not affected much by ionic strength, so approved additives can be used. However, it is not limited to this. This application also applies to powder spray formulations. The proportion of the light compound is not particularly limited, and the compound of the present invention may be used alone, or may be mixed with appropriate additives such as excipients. Examples of formulations are listed below, but other combinations of buffers, preservatives, excipients, tonicity agents, stabilizers, thickeners, various bases (generally referred to as carriers), etc. are also possible. Preferred embodiments of the present invention are shown below, but the scope of the present invention is not limited to these formulation examples.

Example 4 Potassium phosphate 44.028 g and dipotassium phosphate A
Dissolve 1.999 g of N~ruacetylneuramine sodium salt in about 4000 ml of distilled water, and add 100.0 g of N~ruacetylneuramine sodium salt to this.
and 500 mg of benzalkonium chloride were added thereto, and the mixture was made up to 5000 mj7 with distilled water. Next, this liquid was filtered with a membrane filter and dispensed into dedicated containers in 10 ml portions. This preparation has a single spray amount of 0.05 mβ, a single dose of 5i 1 mg, and a pH of 5.0 to 6.0 (0.0
67M phosphate buffer).

Example 5 2.787 g of acetic acid and 37.208 g of sodium acetate were dissolved in about 4000 +++ f of distilled water, and 50.0 g of N-acetylneuraminic acid sodium salt and 15.0 g of chlorobutanol were added thereto. 50 in distilled water
00ml. Next, filter this liquid through a membrane filter.
The mixture was filtered and dispensed into special containers in 10 ml portions. This preparation is administered once at a spray volume of 0.05 mj7 It 0.5
mg, and the pH is 5.0 to 6.0 (0.1M acetate buffer).

Example 6 Sodium phosphate dihydrate? 1.765 g and 5.67 g of disodium phosphate in about 4000 m of distilled water.
1, and to this methyl hydroxybenzoate 1.
30 g and rose propyl hydroxybenzoate 0,70
To this, add 400.0 g of N-acetylneuraminic acid sodium salt, and dissolve the entire amount in distilled water.
000mf. Next, this liquid was filtered with a membrane filter, and 10 ml each was dispensed into special containers. Furthermore,
This was sterilized using high-pressure steam according to a conventional method. This preparation has a one-time spray amount of 0.05ff11, a one-time dose of 4 mg, and a pH of 6.0 to 7.0 (0.1M phosphate buffer).

Example 7 Nasal Disc Formulation Sodium N-acetylneuraminate in a diameter of 3II 1m
It was soaked in a round filter paper and dried. One disk contains 0.1 to 50 mg of the compound of the present invention. One crystal is used by attaching it to the inferior nasal turbinate mucosa.

Example 8 Formulation of Nasal Disk Agent N-acetylneuraminic acid lithium salt was impregnated into a circular filter paper having a diameter of 3 mm and dried. One disk contains 0.1 to 50 mg of the compound of the present invention. One crystal is used by attaching it to the inferior nasal turbinate mucosa.

Example 9 Potassium phosphate 44.028g and dipotassium phosphate 1
．． 999g and about 4000+nj of distilled water! dissolved in
To this, N-acetylneuraminic acid lithium salt 100°O
g and 500 mg'r of penzurkonium chloride were added, and the total volume was made up to 5000 ml with distilled water. Next, add this liquid to
It was filtered with a membrane filter and dispensed into special containers in 10 ml portions. This preparation has a single spray amount of 0.05 mj7, a single dose of 1 mg, and a pH of 5.0 to 6.0 (0,
067M phosphate buffer).

Example 10 2.787 g of acetic acid and 37.208 g of sodium acetate
was dissolved in about 4000 mj2 of distilled water, 50.0 g of N-acetylneuraminic acid lithium salt and 15 g of chlorobutanol were added thereto, and the solution was further diluted with distilled water to 5000 mj2.
And so. Next, this liquid is filtered with a membrane filter, and 10+y+j! Dispense each into special containers. This preparation is 1
When sprayed 10.05 ml, the dose per dose is 0.5 mg, and the pH is 5.0 to 6.0 (0.1M acetate buffer).

Example 11 71.765 g of sodium phosphate dihydrate and 5.67 g of disodium phosphate were dissolved in about 4000 ml of distilled water, and 1.30 g of methyl parahydroxybenzoate was dissolved in the solution.
g and 0.70 g of propyl parahydroxybenzoate were added and dissolved, 400.0 g of N-acetylneuraminic acid lithium salt was added thereto, and the total amount was diluted to 500.0 g with distilled water.
On+j! Closed. Next, this liquid was filtered with a membrane filter, and 10 ml each was dispensed into special containers. Furthermore, this was sterilized using high-pressure steam according to a conventional method. This preparation has a pH of 6.0 to 7.0 (0.1M phosphate buffer) with a single spray volume of 0.05 ml and a single dose of 4 mg.

Example 12 Formulation of Nasal Disk Agent Potassium N-acetylneuraminic acid salt was impregnated into a circular filter paper with a diameter of 3 mm and dried. (2) The disk contains 0.1 to 50 mg of the compound of the present invention. Apply one piece of this product to the inferior nasal turbinate mucosa.

Example 13 44.028 g of potassium phosphate and 1,999 g of dipotassium phosphate were mixed with about 4000+y+j of distilled water! N-acetylneuraminic acid potassium salt l Q
0. Add Og and 500mg of penzalphnium chloride, and add 5000mji with distilled water! Closed. Next, this liquid was filtered with a membrane filter, and 10 mji!
Dispense each into special containers. This formulation has a single spray amount of 0.05
mj! The total dose is 1 mg, and the pH is 5.0~
6.0 (0,067M phosphate buffer).

Example 14 2.787 g of acetic acid and 37.208 g of sodium acetate
and 50.0 g of N-acetylneuraminic acid potassium salt and 1 chlorobutanol were dissolved in about 4000 ml of distilled water. Add Og and further add distilled water for 5 minutes.
000mA. Next, this liquid was filtered with a membrane filter, and 10 mj! Dispense each into special containers. This formulation has a single spray amount of 0.05+nj! One-time administration to,
5 mg, and the pH is 5.0 to 6.0 (0.1M acetate buffer).

Example 15 71.765 g of sodium phosphate dihydrate and 5.67 g of disodium phosphate were added to about 4000 ml of distilled water.
1.3 of methyl parahydroxybenzoate
0 g and 0.70 propyl parahydroxybenzoate
400.0 g of N-acetylneuraminic acid potassium salt was added thereto, and the total volume was made up to 5000 ml with distilled water. Next, this liquid was filtered with a membrane filter and dispensed into dedicated containers in 10 ml portions. Furthermore, this was sterilized using high-pressure steam according to a conventional method. This preparation has 1
If the amount of spraying is 0.05 ml, the amount per dose will be 4 g, and the pH will be 6.0 to 7.0 (0.1 M phosphate buffer).

Example 16 Formulation of Nasal Disk Agent N-acetylneuraminic acid barium salt was impregnated into a circular filter paper having a diameter of 3 mm and dried. One disc of this product containing 0.1 to 50 mg of the compound of the present invention is applied to the inferior nasal turbinate mucosa.

Example 17 44.028 g of potassium phosphate and 1.999 g of dipotassium phosphate were dissolved in about 4000+y+1 of distilled water, and 100 g of barium N-acetylneuraminic acid salt was dissolved therein.
．． 0 g and 500 mg of benzalkonium chloride were added, and the volume was made up to 5000 ml with distilled water.

Next, this liquid was filtered with a membrane filter, and
The mixture was dispensed into special containers in amounts of mj2. This formulation has a single spray amount of 0.
．． 05+nj! The total dose is 1 mg, and the pH
5.0 to 6.0 (0,067M phosphate buffer).

Example 18 2.787g of acetic acid and 37.208g of sodium acetate were mixed with about 4000+y+j! of distilled water! Dissolved in this, N-
50.0 g of acetylneuraminic acid barium salt and 1 chlorobutanol 5. Add Og and add distilled water i:r
50 Q Omll. Next, this liquid was filtered with a membrane filter, and 10 ml each was dispensed into special containers. This preparation has a spray volume of 0.05 mft, a single dose of 0.5 mg, and a pH of 5.0 to 6.0 (0.1 M acetate buffer).

Example I9 71.765 g of sodium phosphate dihydrate and 5.67 g of disodium phosphate were added to about 4000 ml of distilled water.
1.3 methyl hydroxybenzoate dissolved in this
0 g and 0.70 propyl parahydroxybenzoate
400.0 g of barium N-acetylneuraminic acid salt was added thereto, and the total amount was made up to 5000 mf with distilled water. Next, this liquid was filtered with a membrane filter, and 10 mβ portions were dispensed into special containers. Furthermore, this was sterilized using high-pressure steam according to a conventional method. This preparation has a single spray volume of 0.05 ml, and a single dose of 4 mg.
The pH becomes 6.0 to 7.0 (0.1M phosphate buffer).

Example 20 Formulation of nasal disc preparation N-acetylneuraminic acid magnesium salt with a diameter of 3 mm
It was soaked in a round filter paper and dried. One disk contains 0.1 to 50 mg of the compound of the present invention.

One crystal is used by attaching it to the inferior nasal turbinate mucosa.

Example 21 44.028 g of potassium phosphate and 1.999 g of dipotassium phosphate were mixed with about 4000 mj of distilled water! 10% of N-acetylneuraminic acid magnesium salt is dissolved in
Add 0.0 g and 500 mg of benzalkonium chloride,
Further, it was adjusted to 5000 mf with distilled water.

Next, this liquid was filtered with a membrane filter, and
Each ITll was dispensed into a dedicated container. This preparation has a single spray volume of 0.05 mft, a single dose of 1 mg, and a p
H5.0 to 6.0 (0,067M phosphate buffer).

Example 22 2.787 g of acetic acid and 37.208 g of sodium acetate
was dissolved in about 4000 ml of distilled water, 50.0 g of N-acetylneuraminic acid magnesium salt and 15.0 g of chlorobutanol were added thereto, and further dissolved with distilled water for 500 ml.
It was set to 0mj7. Next, filter this liquid through a membrane filter.
The mixture was filtered and dispensed into special containers in 10 ml portions. This preparation has a single spray amount of 0.05 + t + 1, and a single dose of 0.5
mg, and the pH is 5.0 to 6.0 (0.1M acetate buffer).

Example 23 71.765 g of sodium phosphate dihydrate and 5.67 g of disodium phosphate were added to about 4000 ml of distilled water.
1.3 methyl hydroxybenzoate dissolved in this
0 g and 0.70 g of propyl parahydroxybenzoate were added and dissolved, and 400.0 g of N-acetylneuraminic acid magnesium salt was added thereto, and the total amount was diluted with distilled water for 50 g.
000n+j7. Next, this liquid was filtered with a membrane filter, and 10 ml each was dispensed into special containers. Furthermore, this was sterilized using high-pressure steam according to a conventional method. This formulation has a single spray amount of 0.05mji! as a single dose of 4
mg, and the pH is 16.0 to 7.0 (0.1M phosphate buffer).

Example 24 500.0 g of lactose and 500.0 g of N-acetylneuraminic acid sodium salt were mixed in a powder mixer, and then 40 mg of mixed powder was filled into gelatin capsules in a capsule filling machine. The drug was manufactured. The compounds of the invention are administered into the nasal cavity using a dedicated powder nebulizer;
In this case, the one-time dose of the compound of the present invention was 2 omg.

[Brief explanation of drawings]

FIG. 1 is a diagram showing the effect of N-acetylneuraminate of the present invention on suppressing nasal mucosal vascular permeability.

Claims (4)

[Claims] (1) General formula; ▲There are mathematical formulas, chemical formulas, tables, etc.▼ A nasal mucosal vascular permeability inhibitor containing a compound shown as an active ingredient. (However, when n is 1, Z is lithium, potassium, sodium, ammonium or organic ammonium, and n
When is 2, Z represents calcium, barium or magnesium. ) (2) The nasal mucosal vascular permeability inhibitor according to claim (1), wherein the agent is administered to the nose as a spray, a drop, or a disc. (3) The agent for suppressing nasal mucosal vascular permeability according to claim (2), which comprises an effective amount of the compound and a pharmaceutically acceptable carrier. (4) The nasal mucosal vascular permeability inhibitor according to claim (3), wherein the carrier is a stabilizer, a preservative, a buffer, an isotonic agent, or an excipient.