JPS5946927B2 - antiallergic agent - Google Patents

Description

[Detailed description of the invention] The present invention relates to a novel antiallergic agent.

More specifically, the present invention relates to an antiallergic agent containing the glycoprotein WENAC as an active ingredient.

Many anti-allergic agents have been known to date, but there are anti-allergic agents that specifically suppress the production of immunoglobulin E (IgE) antibodies, which are the main cause of immediate allergies, such as atopic asthma. At present, little is known about it and there are no drugs in practical use, and the appearance of such a drug has been eagerly awaited.

The antiallergic agent of the present invention that specifically suppresses the production of gE antibodies is extremely innovative.

That is, the antiallergic agent of the present invention specifically suppresses the production of IgE antibodies.

In addition to this effect, it blocks the attachment of IgE antibodies to target cells and suppresses the release of histamine upon antigen invasion.

That is, it suppresses the release of histamine based on antigen-antibody reactions.

Therefore, the antiallergic agent of the present invention is extremely useful in particular for the treatment and prevention of allergic diseases based on antigen-antibody reactions, that is, immediate allergies typified by atopic asthma.

The present invention was first filed by the inventors in Japanese Patent Application No. 50-122197.
This invention corresponds to a further development of the invention for which a patent application was filed in No. 52-47909.

That is, the invention of the previous Japanese Patent Application No. 122197/1983 is based on the antiallergic, anticancer, and immunostimulatory effects on lung cells and lymphocytes obtained by culturing a strain belonging to nonpathogenic aerobic Corynebacterium. The invention relates to a method for producing a water-insoluble culture having the following characteristics.

However, since the resulting culture (bacterial cells) is insoluble in water, when injected subcutaneously as a suspension, induration occurs similar to when BCG is injected subcutaneously, making continuous administration impossible. In addition, there were drawbacks such as strong toxicity, so
In reality, it has been difficult to use it as a medicine.

As a result of further research, the present inventors have discovered a water-soluble active substance that can be practically used as a pharmaceutical product from a culture (bacteria) of non-pathogenic aerobic Corynebacterium that has various useful pharmacological effects. We succeeded in separating and extracting it.

They confirmed that this separated extract was a single substance consisting of proteins, sugars, and nucleic acids, and named it WENAC.

That is, the glycoprotein WENAC of the present invention not only specifically suppresses the production of IgE antibodies, which are the main cause of immediate allergies, and has a strong antiallergic effect, but also has a strong antiallergic effect on the raw material culture (
It is also significantly less toxic than bacterial cells.

Since this glycoprotein WENAC is water-soluble, it can be administered continuously by subcutaneous injection, and can be used in accordance with desensitization therapy, which is important as a treatment for allergic diseases, and can be used practically as a medicine. I can do it.

First, to explain the method for producing the substance of the present invention, in the invention of Japanese Patent Application No. 50-122197, the bacteria used are all strains belonging to non-pathogenic aerobic Corynebacterium, such as Corynebacterium equii IIDKo- The target product was obtained by culturing 85 strains aerobically, and in the present invention, the target product is used as a starting material.

The strains belonging to non-pathogenic aerobic Corynebacterium used in the present invention include Corynebacterium quii (
Corynebacterium equi)IIDK
o-s5, Corynebacterium pyogenes (Cory
nebact, erium pyogenes) II
Corynebacterium xerosis
) I ID 53-1, Corynebacterium renale (Corynebacterium erium r.
enale) I I DUt, sunomiya-Us
An example is hi.

All of these bacterial strains can be easily obtained by those skilled in the art, and are stored as standard strains at the Institute of Medical Science, the University of Tokyo (JFCCCat, alogueo
f Cult, ures, 1962).

A particularly preferred strain is Corynebacterium equi IIIDK.

-85.

These non-pathogenic aerobic Corynebacterium cultures (bacterial cells) can be obtained by culturing aerobically using a culture method commonly used for aerobic culture of Corynebacterium.

The raw material culture thus obtained (bacterium 9, that is, the culture (bacteria) obtained by aerobically cultivating a non-pathogenic aerobic strain belonging to the genus Corynebacterium, is preferably treated at a low temperature. , the resulting crushed product is treated with ultrasonic waves to form a precipitate in the separated aqueous solution of the active ingredient within the range of pH 3.70 to 3.75, and this precipitate is converted into an approximately neutral aqueous solution and subjected to dialysis. The water-soluble active substance, the glycoprotein WENAC, is obtained by subsequent lyophilization if desired.

Although the details of the constituent components of the glycoprotein WENAC of the present invention are unknown, it is known to be a glycoprotein substance containing a protein part, a sugar part, and a nucleic acid part in a certain ratio.

The glycoprotein WENAC of the present invention has the following physical and chemical properties.

a) Ratio of protein part to sugar part: The ratio of protein part/sugar part calculated from the protein content by the copper folin method and the sugar content by the phenol sulfur method is 3.9.
70.52 to 5.510.41 or 7.50 to 13.4
It is 2.

The protein content by the copper foreign method was determined by calf serum albumin (
The sugar content determined by the phenol-sulfuric acid method was determined as a converted value of glucose (BSA).

b) Nucleic acid moiety: The ratio of optical density (OD) at 260 nm and 280 nm (260 nm/280 nm) is 1.56 to 1.62.

This ratio means the quantitative relationship between the nucleic acid part and the protein part.

(Japan Biochemistry Synthesis Biochemistry Experiment Course 5 Enzyme Research Methods Part 1 Tokyo Kagaku Doujin Publishing Published August 20, 1975 No. 24
- Page 26) C) Isoelectric point: 3.75 to 3.80 The isoelectric point determined by the crystal precipitation method is within the range of 3.5 to 4.1, but when the apex is determined, it is approximately 3.75 to 3.80. It becomes 3.80.

d) Molecular weight: Elution curve in Sephadex G200 gel: As shown in FIG.

Elution curve in Urtogel AcA34: Figure 1-B
As shown below.

e) Elemental analysis: The relative ratio of CHN calculated from the elemental analysis value is as follows.

C71 section HIO Chi N19 section The elemental analysis value varies depending on whether dialysis is performed using physiological saline during the manufacture of this product, etc., but the relative ratio of CHN calculated from the elemental analysis value is as shown above. constant.

f) Infrared absorption spectrum: As shown in FIG. 5, it has characteristic absorption.

g) Ultraviolet absorption spectrum: As shown in Figure 6, it has an absorption maximum and an absorption minimum.

h) Color and shape of the substance: The aqueous solution (0.5%) of slightly yellowish white powdered milk crystal is a light yellowish brown, clear to slightly cloudy liquid.

i) Distinction between basic, acidic and neutral: An aqueous solution of quartz (0.5%) exhibits a pH of 6.2 to 7.8.

j) Color reaction: Viillet reaction (protein qualitative reaction), Annexon reaction (sugar qualitative reaction) positive k) Specific rotation: [α']D-50,32±9.44 (C20.2 .water)
υ Constituent Amino Acids: The following amino acids are confirmed by hydrolysis with 6N hydrochloric acid.

Alanine, glutamic acid, glycine, leucine.

Aspartic acid, valine, lysine, threonine.

Arginine, serine, florine, isoleucine.

Phenylalanine, histidine, methionine.

Tyrosine m) Constituent sugars: At least the following sugars are identified by hydrolysis of 2.5N trifluoroacetic acid.

Production method of rehose, mannose, galactose, glucose raw material culture (box) Corynebacterium equii IIDKo-85 as culture medium (meat extract log, peptone 10g, glucose 10g)
G2 1 g of common salt and 1000 yd of distilled water (adjusted to pH 7.4 with an aqueous sodium hydroxide solution) were used to culture at 37° C. for 5 days.

Next, the flow rate was 50rrII! The mixture is centrifuged at 10,000 revolutions per minute, and the precipitate (box 9 is washed three times with physiological saline) is used as a raw material culture (bacteria).

The yield of cultured cells is 17.0g per 10t of culture medium used.
(wet weight).

Production Example of WENAC 1 50 g of the raw material culture (bacterial cells) is sufficiently washed with physiological saline, subjected to low temperature treatment at -20°C for 48 hours or more, and then crushed by a vibrogen mill.

The crushed material is suspended in an appropriate amount of water or aqueous sodium hydroxide solution, and subjected to 20 kilocycle (KC) sonication in ice water six times for 10 minutes each, followed by shaking extraction.

150 g of the obtained extract is centrifuged at 1105000X for 60 minutes to remove destroyed bacterial cell residue and insoluble matter.

This supernatant liquid was adjusted to a concentration equivalent to 25 cm of bacterial cells at 8-10°C.
The precipitate is aged for at least 48 hours at a pH of 3.70 to 3.75 using IN hydrochloric acid.

The precipitate was collected by centrifugation, adjusted to pH 7.2, and dissolved in water.
Perform dialysis overnight and correct the pH to 7.0 to 12,000.
After centrifugation at xg for 60 minutes, obtain a supernatant (extract).

This product is then freeze-dried to obtain the glycoprotein WENAC.

Yield: 500 ~ (dry weight). Production example of WENAC 2 After 152 g of the raw material culture (bacterial cells) was low-temperature treated at -20°C for 48 hours, it was suspended in distilled water together with glass beads with a diameter of 0.1 m, and Vibrogen Cell Mill (VCH- 1)
The microbial cells were disrupted by performing the treatment 10 times at 450 rpm for 30 minutes.

The supernatant was then centrifuged at 1100 Orp for 10 min, followed by 1 20 kilocycle (KC) sonication in ice water.
5 minutes, 3000 rpm for 20 minutes, 16000 rpm
Centrifuge at rpm (20,000 G) for 40 minutes to remove destroyed bacterial cell residue and insoluble matter.

This supernatant liquid was made into a concentration equivalent to 25 cells of bacterial cells, and was incubated at 4°C.
Adjust the pH to exactly 3.70 using hydrochloric acid and leave overnight to ripen the precipitate.

The precipitate was collected by centrifugation at 5000 rpm for 30 minutes, adjusted to pH 7.0 with INN nanatrirum hydroxide, dissolved in distilled water, and dialyzed overnight against distilled water at 4°C and then against physiological saline for 2 days. Adjust to pH 7.0, 16
After centrifugation at 2,000 rpm (2,000 G) for 40 minutes, the supernatant was filtered through a membrane filter (Millipore HA) to obtain 500 ml of extract.

Next, this extract was freeze-dried to obtain the glycoprotein WENAC.
get.

A Physical and chemical properties of this extract The appearance of this extract is a pale yellow and clear liquid.

When the pH of this extract is adjusted to 3.75 to 3.80, precipitation occurs.

According to the above manufacturing example, 13 samples (lots) were prepared (extract liquid).
were individually adjusted, and for each sample expressed with AI = "13, *1 protein content by copper folin method, *2 sugar content by phenol-sulfuric acid method, ratio of protein part / sugar part calculated from these contents, *3 Regarding the nucleic acid part 2
The results of measuring the ratio of optical density (OD) at 60 nm and 280 nm (260 nm/280 nm) and the isoelectric point by the precipitation method are shown below.

*l Protein content (copper foreign method) Add water to this extract to make a 10-fold dilution, and dilute it with 0.6y.
Add solution C 3- to d and allow to react at room temperature for 10 minutes.

After the reaction, the phenol solution (commercially available product diluted with water) was 0.37
After adding nl and leaving it at room temperature for 30 minutes, 500nm
Measure the OD of.

Calibration curve by BSA 0.2-1.077%'/m/
! The protein content of the test solution (B
SA conversion value).

Liquid A: 0. Add sodium carbonate to the IN sodium hydroxide solution to make the concentration 2.

Solution B: Add copper sulfate to 10% sodium citrate solution to make the concentration 0.5.

Solution C: 50d of solution A: 1m of solution B (50:1) *2 Sugar content (
Phenol sulfuric acid method) Add water to this extract to make a 10-fold dilution, and
Immediately add 1 rnl of 5-thiphenol solution and 5 ml of sulfuric acid to the solution.

After leaving it for 20 minutes, measure the OD at 490 nm.

Calibration curve with glucose aqueous solution 0.02-0.21n
The sugar content (glucose equivalent value) of the test solution is determined from the calibration curve prepared within the concentration range of fI/d.

*3 Add water to the nucleic acid partial extract to dilute it to an appropriate concentration, and analyze it at 260 nm.
and 280 n r, the optical density (OD) at 1 is measured and the ratio thereof is determined.

B Physical and chemical properties of glycoprotein WENAC ■ The extract A I = 13 is individually dispensed into 2- vials and freeze-dried to obtain glycoprotein WENAC.

For each specimen expressed as Al to I3, the protein content by the copper Folin method, the sugar content by the phenol-sulfuric acid method, the ratio of protein part/sugar part calculated from these contents,
The results of measuring the ratio of optical density (OD) at 260 nm and 280 nm (260 nm/280 nm) and isoelectric point by precipitation method for the nucleic acid portion are shown below.

For the measurement of copper, protein content using the 7-ohlin method, sugar content using the phenol-sulfuric acid method, and nucleic acid content, add 2 ml of water to each vial of the sample. After adding and dissolving WENAC, the above *l, *2. *It was carried out in the same manner as in 3.

In addition, the sample A14 was obtained by adding water 2- to 13.8277v of A5 WENAC (freeze-dried product) and dissolving it in the same manner as above.

(2) The results of measuring the relative ratio and specific rotation of CHN calculated from the elemental analysis values for each of the above specimens (WENAC) are shown below.

Specific optical rotation was measured by dissolving 10 μm of each sample in water, adding an appropriate amount of urea to make a total volume of 5 μg, and centrifuging the solution if necessary.

■ Molecular weight: The elution curve of this product in Sephadex G200 gel shows a single peak as shown in Figure 1.

Head volume is 3.0 x 60 cm, developing liquid is 0.01
The flow rate is 20.8 rnl/h with M phosphate buffer.

The vertical axis shows the optical density (OD) at 280 nm, and the horizontal axis shows the elution amount.

The elution curve of this product in Ultrogel AcA34 is shown in Figure 1-B.

Column size is 2.8X84z, developing solution is 0.01 MI
J acid buffer (containing 0.85% saline) with a flow rate of 14
．． 0m/hour.

The vertical axis shows the optical density (OD) at 280 nm, the horizontal axis shows the elution amount (-),
160,000), indicates the elution position of twinbin trypsin inhibitor (molecular weight: 21,700).

■ Infrared absorption spectrum: The infrared absorption spectrum of this product measured by the potassium bromide tablet method has characteristic absorption as shown in Figure 5.

■ Ultraviolet absorption spectrum The ultraviolet absorption spectrum of this product has an absorption maximum and an absorption minimum as shown in Figure 6.

■ Color and shape of substance: Slightly yellowish white powder.

The aqueous solution (0.5%) of this product is a pale yellow-brown clear to slightly cloudy liquid.

■ Distinction between basic, acidic, and neutral: The aqueous solution (0.5%) of this product exhibits a pH of 6.2 to 7.8.

■ Color reaction: Viillet reaction (protein qualitative reaction) positive.

That is, when adding 0.57 nl of sodium hydroxide reagent to 0.5 me aqueous solution (0.5%) of this product to make it alkaline, and then adding 3 drops of copper acid solution (1→l 00 ), the liquid turned purple. exhibits.

Anthrone reaction (sugar qualitative reaction positive).

That is, anthrone test solution 1- is added to 0.5 liters of an aqueous solution (0.5%) of this product, and when heated in a water bath, the solution takes on a green color.

■ Constituent amino acids: The following amino acids are identified by hydrolysis with 6N hydrochloric acid.

That is, a 6N hydrochloric acid solution containing 10 mg of this product was placed in an ampoule, degassed and sealed, and then heated and hydrolyzed at 110°C for 24 hours.
The following amino acids are identified when analyzed by automatic amino acid analysis.

Alanine, glutamic acid, glycine, leucine, aspartic acid, valine, lysine, threonine, arginine,
serine, proline, inleucine, phenylalanine,
Histidine, methionine, tyrosine, [Phase] Constituent sugars: 2.5N) At least the following sugars are confirmed by hydrolysis of lifluoroacetic acid.

That is, 10 ml of a 2.5 N refluoroacetic acid solution containing 10 mg of this product is placed in an ampoule, degassed and sealed, heated and hydrolyzed at 100° C. for 7 hours, and then heated and dried.

Then, it was dissolved in 0.25% of water, and Amberly) 120(
H) and Amberlite 4B (-COO-), heat-dry the sugar eluted portion, dissolve it in 0.1 ml of water, and analyze it using a reducing sugar automatic analysis system, at least the following sugars are confirmed.

Biological properties of ribose, mannose, galactose, glucose C glycoprotein WENAC ■ Immunoglobulin E (IgE) antibody production inhibitory effect 2
μg of DNP-OA was suspended in physiological saline together with aluminum hydroxide gel and administered intraperitoneally to BALB/C mice, and then 600 μg of WENAC was administered intraperitoneally on the 2nd and 7th day.

Five days after immunization, the serum from the mouth on days 10, 15, 20, 25, and 30 was pooled and sensitized to the dorsal skin of SD rats, and 4 hours later, antigen DNP containing 0.5 Evans blue was injected from the tail vein. -BSA solution was injected, and IgE activity was measured based on the dye leaching reaction that occurred.

DNP-OA sensitized BALB/C mice were divided into three groups, and the mice of the first group (vertical axis in Fig. 2) received WENAC, and the mice of the second group received WENAC.
Group mice (vertical axis △ in Figure 2) were sensitized with killed anaerobic Corynebacterium parvum cells, and group 3 mice (vertical axis 〇 in Figure 2 were sensitized with physiological saline as a control). Day 2 and 7
Inoculated on day → - day.

The results are shown in FIG. In FIG. 2, the vertical axis shows the IgE antibody titer, and the horizontal axis shows the number of days after DNP-OA sensitization.

From FIG. 2, it is clear that the WENAC of the present invention specifically suppresses IgE antibody production, which is the main cause of immediate allergy.

■ Inhibition of passive anaphylaxis (PCA) SD rats were treated with the WENAC of the present invention in advance, and 2 hours later they were inoculated with anti-DNP (hapten) IgE antibody, resulting in a typical <IgE antibody reaction. The results of the passive anaphylaxis reaction (PCA) are shown in Table 1: As shown in Table 1, the WENAC of the present invention inhibits the attachment of IgE antibodies to target cells (mast cells, salophil cells, etc.). block and inhibit histamine Ω release during antigen invasion.

■ IgE antibody adhesion blocking effect on target cells and I
The effect of blocking gE antibody adhesion to target cells was more pronounced when treated with the WENAC of the present invention before IgE antibody sensitization than when treated after sensitization.

The results are shown in Table I-B.

In the same table, the processing amount of WENAC of the present invention is 1.0'InI
! The PCA value is expressed by the amount of extracted dye.

The above experimental results demonstrate that the WENAC of the present invention blocks the binding of IgE antibodies sensitized to target cells to target cells, regardless of the type of antigen, and suppresses a wide range of subsequent allergic reactions.

■ Enhancement of anti-hapten IgG antibody production in 7-week-old BAs
LB/C mice were intravenously injected with WENACO017IIg of the present invention, and 7 and 14 days later, they were intravenously sensitized with 100 μg of DNP-OA.

Furthermore, after 3 days, the number of antibody-producing cells in the lung cells was determined by DNP-
It was measured by Cunningham's indirect method using BSA-conjugated sheep red blood cells.

The results are shown in Table H.

It is clear that the WENAC of the present invention enhances the production of anti-hapten IgG antibodies, as shown in Table 2.

Therefore, it is clear that the WENAC of the present invention has an immunostimulatory effect, and even though the ability to produce antibodies other than allergic antibodies is promoted by administration according to the basic principle, it does not decrease.

■ Common antigenicity Common antigenicity was investigated by in-gel precipitation reaction (Ophthalony method).

Using anti-Corynebacterium equii rabbit serum and anti-tuberculosis rabbit serum and using the WENAC of the present invention and tuberculin protein solution (TA2) adjusted to 1101 n/ml as antigens, an in-gel precipitation reaction was performed by the Ophthalony method. .

The results are shown in FIG.

In the figure, the left figure shows anti-Corynebacterium rabbit serum (top ○
) against tuberculin protein hemolysis (TA2) (bottom left ○
) and WENAC of the present invention (lower cloth ○) are shown.

The right figure shows the common antigenicity of TA2 (bottom left ○) and WENAC of the present invention (bottom cloth ○) with anti-tuberculosis rabbit serum (top ○).

From the sedimentation line shown in FIG. 3, it is clear that WENAC of the present invention has common antigenicity with tuberculin protein.

■ Anti-complement effect After reacting SD rat serum with various concentrations of WENAC of the present invention at 37°C for 1 hour, residual hemolytic activity of complement was measured using sheep red blood cells sensitized with anti-sheep red blood cell antibodies. did.

The results are shown in FIG.

In the figure, the horizontal axis shows the added amount (μg) of WENAC of the present invention, and the vertical axis shows the inhibition rate (%) of the sheep hemolytic reaction.

As is clear from FIG. 4, it was confirmed by the inhibition rate of the hemolytic reaction of sheep red blood cells that the WENAC of the present invention has a significant anti-complement effect.

This anti-complement effect has also been confirmed by reducing complement levels in the serum of animals treated with the WENAC of the present invention.

Therefore, the WENAC of the present invention is considered to suppress non-specific allergic reactions.

■ Polymyxin B and Compound 4
Inhibitory effect of 8/80 on histamine release from rat mast cells a)) Squirrel buffer TACM [0,02M) Squirrel, 0
-12M Na C1t O-003MK CI, 0
．． 1% glucose (1)t O-001M0-0O1
and 0-4 rn M Mg Cl 2 , pH 7-
2XIO rat intraperitoneal mast cells suspended in W
After acting with ENAC for 30 min at 37°C, Polymynin B or Com1pound 4
The table below shows the inhibition rate of histamine release from mast cells when reacted with 8/80.

As is clear from the table, histamine release by Polymyxin B was more significantly suppressed than WENACl 2.5 μg/Tll (more than 40), and suppression of histamine release by Compound 48/80 was observed at WENAC 50 μg/Tll or more.

b) Polymyxin B and Compo in a)
WENA showed sufficient effect on both und48/80
Using 100 μg/− of C, the influence of pretreatment time on rat mast cells on the inhibitory effect of WENAC on histamine release was confirmed in the same experimental system as in a).

The results are shown in the table below. As is clear from the table, P
o lymyx in B and Compound 48
/ 80, an inhibition rate of 50 or more was observed with WENAC pretreatment for 15 minutes or more.

■ Inhibitory effect on histamine release from rat mast cells by simultaneous use of concanavalin A and phosphatidylserine Intraperitoneal obesity of 2 x 105 rats suspended in Tris buffer TACM (same as test example ■a)) The table below shows the rate of histamine release from rat mast cells when the cells were treated with concanavalin A (conA), phosphatidylserine (PS) and/or WENAC at 37°C for 30 minutes.

As is clear from the table, con A in rat mast cells
It was confirmed that only when IOOpg/me and 50 μg/- of PS were applied simultaneously, histamine was significantly released at a level of 67, but in other cases, almost no release was observed.

It was confirmed that the release of histamine due to the simultaneous use of conA and PS was significantly suppressed from 67% to 10% by the use of WENA Clooo μg/ml. Culture (bacterium), dd
The acute toxicity when administered intraperitoneally to N mice (body weight 20 g) is as follows.

In addition, since the raw material culture (bacteria cells) is insoluble in water, it was administered as an aqueous suspension.

WENAC of the present invention LD5o: 20■ or more/mouse raw material culture (bacterial cells) LD5o: 5■/mouse or more From the experimental results, the WENAC of the present invention is the raw material culture (bacterial cells)
It has been shown that the toxicity is significantly lower than that of

In addition, no abnormalities were observed in the mouse leukocyte reduction test and mouse weight loss test according to the general test method of the WENAC "biological product standards" of the present invention.

When using the WENAC of the present invention as a medicine, it is used in a form convenient for obtaining the desired medicinal effect.

WENAC may be a medicament in its pure state, but it may also be provided in a composition as a mixture with pharmaceutically acceptable diluents and/or other pharmacological agents in accordance with pharmaceutical practice.

The medicament of the invention can be applied orally or parenterally.

Therefore, the medicament of the present invention can be appropriately administered in a form for oral or parenteral administration.

For example, powder. 2 tablets of granules, sugar-coated tablets, capsules, pills,
Suppositories, suspensions, solutions, emulsions, injections, and aerosols.

The dosage of the medicament of the present invention has a sensitivity difference of 2 years and gender.

Body weight, administration method, 2 administration intervals, medical condition 2 physical condition.

It varies depending on various factors such as the nature of the pharmaceutical preparation and the type of preparation.

Therefore, doses less than the minimum dosages indicated below may be sufficient, and in some cases it may be necessary to administer more than the dosages indicated below.

Dosage for adult patients is Long ~ 500 per day
It is μg.

Prescription example l (injection) WENAC (freeze-dried product) of the present invention at 57nf! /77
Z71! and further diluted with physiological saline to 100%
Dilute twice and fill 1 ml each into ampoules.

The standard usage of this injection solution is to subcutaneously inject the same amount of 0.1 to 0.8 mA once or twice a week in accordance with normal desensitization therapy.

Prescription example 2 (injection 9 WENAC (lyophilized product) of the present invention 5.0 per rnl
An aqueous solution containing μg and mannitol 1107n, 1
Fill ml into vials and lyophilize.

[Brief explanation of drawings]

Figure 1 shows WENAC's Sephadex G200 of the present invention.
The elution curve in the gel is shown. FIG. 1-B shows the elution curve of WENAC in Ultrogel AcA34. FIG. 2 shows the IgE antibody production suppressing effect of WENAC of the present invention. FIG. 3 shows the common antigenicity of WENAC, and FIG. 4 shows the anti-complement effect. 2 to 4 show the effects of the present invention. FIG. 5 shows the WENACQ infrared absorption spectrum. FIG. 6 shows the ultraviolet absorption spectrum of WENAC.

Claims (1)

[Claims] 1. An anti-allergic agent containing the glycoprotein WENAC as an active ingredient.