JPH11228432A - Influenza vacuum action potentiator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the subject potentiator be including 'Maoubushisaishinto' to stimulate the immune strength declined as a result of combined use of an influenza vaciine and have effect of raising the action of the vaccine. SOLUTION: This influenza vaciine action potentoiator contains 'Maoubushisaishinto' which is prepared by prescription of Ephedra sinica, aconiti tuber, and Asarum Sieboldi in a proportion of 4:1:3 by weight and has been out on the market as a mixed crude drug. In this potentiator, a dried extract afforded by subjecting the above mixed crude drug to extraction with water is usable as the active ingredient. This potentiator is orally administrable, the dialy dose of the 'Maoubushisaishinto' being 100-3.600 mg, pref. about 300-2,000 mg, more pref. about 500-1,500 mg. normally about 1,200 mg, in terms of the dried extract.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to an influenza vaccine action enhancer containing a Chinese herbal medicine-Mao-bushi-saishin-to.

[0002]

BACKGROUND OF THE INVENTION Influenza, also called epidemic cold, is the most important respiratory viral disease and is often pandemic. Since there are several types of influenza viruses, it is said that anticipating the epidemic in the winter and inoculating it at least two weeks before the epidemic will have a protective effect. However, the effects are unsatisfactory and lack permanence. Furthermore, even if a pandemic can be expected, vaccine production requires complicated operations and time, and production cannot keep up. In addition, the immune function of a living body gradually decreases with age. Getting flu can be fatal for older people. Therefore, a drug that enhances the action of the influenza vaccine has been demanded.

[0003] The inventors of the present invention have studied the effects of Kampo medicines on influenza virus (IFV), and have completed the present invention based on the finding that mao-bushi-saishin-to enhances the effect of influenza vaccine.

[0004]

A specific embodiment of the present invention is an influenza vaccine action enhancer comprising Mao-bushi-saishin-to.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION Mao-bushi-saibushi-to is also referred to as mao-bushi-shinbashi-to and is listed as one of the main Chinese medicines. The prescription consists of Mao 4, Spicy 3 and Bushi 1. This herbal medicine has general malaise, apathy, mild fever, chills, and has been used for colds and bronchitis.

[0006] Ephedrae herba is a species of Ephedra (Ep
hedraceae) ( E. sinica STAPF) and other stem plants of the same genus. The active principal components are (−)-ephedrine, (+)-pseudoephedrine, (−)-methylephedrine and the like. It is used as an antipyretic and antitussive.

[0007] Aconiti tuber has a root of a Chinese beetle, such as Chinese beetle, and a root attached to the root is a root, but both marketed products are called a root or a head. After salting and heat treatment such as steaming, it is called a gun attachment. It is used in Chinese medicine for the purpose of analgesia, diuresis, intenseness, and promotion of metabolism.

[0008] Hot spice (Asiasari radix) is a dried product of the roots and rhizomes of Usubasaicin or Keirinsaishin, and is used as an antitussive, analgesic, excretory, and diuretic.

Mao-bushi-saishin-to is a prescription of the above-mentioned mao-bushi, spicy spice in a ratio of 4: 1: 3, and is commercially available as a mixed crude drug. The water-based dried extract extracted from this mixed crude drug can be used for preparing the preparation of the present invention.

The influenza vaccine action enhancer of the present invention may be administered from about 14 days before the vaccine to about 3 days after the vaccine, preferably from 8 days before the vaccine to 1 day after the vaccine, more preferably from 6 days before the vaccine to 6 to 8 hours after the vaccine. You can start later. The administration is preferably performed continuously for 2 to 14 days, more preferably for 3 to 8 days, most preferably for 5 to 6 days.

The influenza vaccine enhancer of the present invention can be administered orally, and a water-based dry extract can be administered as it is. Capsules, fine granules, granules, tablets, soft jellies, drinks, dry syrups Agents, lozenges, and the like. In addition, it is possible to ingest commercially available Mao-bushi-saishin-to for drinking.

For the preparation of the above-mentioned preparations, commonly used additives such as excipients, diluents, sweeteners, flavors, corrigents, suspending agents and stabilizers can be used. Excipients include:
Lactose, corn starch, inorganic salts (magnesium aluminate metasilicate, perfiller, etc.), carrageenan, locust bean gum and the like can be mentioned. Examples of the dilution / mixing agent include sterilized purified water, a mixed solution of ethanol and sterilized purified water, and the like.

The sweetener includes reduced maltose, maltose, sugar, xylitol and the like. Flavors include orange flavor, apple flavor, grapefruit flavor, peppermint flavor and the like. Examples of the flavor include ascorbic acid, citric acid, β-cyclodextrin, lecithin and the like. Suspending agents include Neusilin, polyoxyethylene hydrogenated castor oil and the like. Examples of the stabilizer include methyl 4-hydroxybenzoate.

The daily dose of Mao-bushi-saishin-to is 100 to 3600 mg as a dry extract made of water, but it can be increased or decreased according to symptoms, age, physical strength and the like. Preferably, it is about 300 to about 2000 mg, more preferably about 500 to about 1500 mg. Usually about 1,200 mg.

The influenza vaccine which is the object of the present invention is called influenza HA vaccine in Japan, and is produced by culturing various influenza viruses in embryonated chicken eggs and decomposing and purifying virus particles collected from the allantoic fluid. It is a multivalent vaccine containing the obtained protective antigenic substance as a main component (all microorganisms are inactivated using formalin during the purification process). Usually, two subcutaneous inoculations at weekly to monthly intervals provide basal immunity. But,
It may be a live vaccine containing a live attenuated microorganism, which is not currently in practical use, or a component vaccine detoxified so as not to impair the immunogenicity of the toxin.

Pharmacological experiments The effect of mao-bushi-saishin-to to enhance the influenza vaccine action is shown by the increase in antibody titer. Test method Animals: C3H / HeN (Slc) female mice of 1 year old were used. The experimental groups consisted of a group of 0.0 mg, a group of 0.5 mg,
And 5.0 mg administration group, and 5 to 6 mice in each group were used.

Mao-bushi-saishin-to water-dried extract: Mao-bushi-saishin-to was used as an extract prepared as follows. Mao (Ephedrae Herba, Osaka market product), gun attachment (Ac
oniti Tuber, Osaka market product), spicy (Asiasari Radix,
Osaka market product), mixed at a weight ratio of 4: 1: 3, added with 10 times the volume of deionized water, and subjected to reflux extraction at about 100 ° C. for 1 hour. After the extract was filtered through a 100-mesh sieve, it was freeze-dried to prepare a mao-bushi-saishin-to extract. The extract yield was 15%.

Administration method: A dried extract made of mao-bushi-saishin-sui-water is suspended in deionized water to prepare a concentration of 5 mg / ml and 50 mg / ml. 25
Oral administration of 0.1 ml per gram (20 mg / kg / day x 5 times) was performed. In addition, a 50 mg / ml solution was administered to the 5.0 mg group,
Oral administration of 0.1 ml per 200 g (200 mg / kg / day x 5 times) was performed. All the administrations were performed once a day for 5 days using a gastric probe. The same amount of deionized water was orally administered to the control group, which was administered at 0.0 mg, in the same manner.

(1) In the experiment of administration before inoculation, Mao-bushi-saishin-to was orally administered for 5 days before the first vaccination. (2) In the post-inoculation administration experiment, administration of mao-bushi-saishin-to was started from 6 to 8 hours after the first and second vaccinations, and orally administered for 5 days.

Vaccination: Vaccination was carried out in '96.
Influenza HA vaccine "BIKEN HA" (A / Yamagata / 32/89 (H1N1) strain, A Wuhan / 359/95)
(H3N2 strain, B / Mie / 1/93 strain)
The animals were inoculated subcutaneously. Three weeks later, the same amount of the vaccine was subcutaneously administered again.

[0021] In each experiment, 6 of the first vaccination was performed.
Days before and the day before and 6 days after the second vaccination,
Blood was collected after 4 weeks, 4 weeks, and 8 weeks, and the change in anti-vaccine-IgG antibody titer in the serum was measured using an ELISA method.

ELISA method: 0.05M Carbonate Bu
Influenza virus vaccine diluted to the optimal concentration with ffer (pH 9.6)
Dispense μl and leave it at 37 ° C for 2 hours to immobilize it. 0.05
PBS (-) containing% Tween 20 (hereinafter Tw-PBS)
After washing with, 100 µl of the test serum diluted with PBS (-)
And allowed to stand overnight at 4 ° C. to react with the antigen-specific antibody. After washing with Tw-PBS, Peroxi diluted to the optimal concentration
dase-Rabbit anti-mouse IgG (H + L) (ZYM
ED, Code No. 61-6520) Add 100 μl, 37 ° C
And react for 1 hour. After washing with Tw-PBS, 100 μl of the substrate solution is added, and the reaction is carried out at room temperature for 30 minutes while shielding from light. Substrate solution is 40mg o-phenylene diamine and 40μ
l 30% H ₂ O ₂ was added to 0.01M Citrate Buffer (pH
5.0) to make 100 ml. 2N H ₂
The reaction was stopped by adding 100 μl of SO ₄ and then MI
4 at CRO PLATE READER (TOSOH)
The absorbance at 92 nm was measured. The absorbance value of the test serum was defined as the difference from the well containing no serum, corrected based on the standard value of the control serum on the same plate, and the anti-vaccine-IgG antibody titer was determined using the dilution ratio of the serum. And

Experimental Results (1) Table 1 shows the effect of mao-bushi-saishinto on IgG antibody titers after influenza vaccination by administration before influenza vaccination (the numerical values indicate IgG antibody titers).

[0024]

[Table 1] Pre-vaccination administration 1st vaccination 2nd vaccination 2nd vaccination 2nd vaccination 2nd vaccination 2nd vaccination 6 days after day 6 days after 2 weeks after 4 weeks after 8 weeks Number of specimens 15 14 14 14 14 13 0.0mg 1 5110 9067 14742 18635 12104 8426 Administration 2 6052 (Control) 3 4343 4 4163 15178 26236 24881 25444 21457 5 5411 12261 32373 41093 65497 23553 Average 5016 12169 24450 28203 34348 17812 SD 696 2496 7308 9465 22689 6692 0.5mg 1 4273 33725 44 336 86 198 49782 29549 Administration 2 3772 59851 70744 59252 45617 28466 3 5320 18703 33993 22751 17310 4 3222 57001 100 118 87925 67157 47552 5 2971 15992 36613 27543 19815 34771 Average 3912 37054 57161 56734 39936 35085 SD 836 18488 25113 27764 18905 7584 5.0mg 74734 122981 102570 94943 80544 Administration 2 7075 28421 24398 30069 25348 20061 3 11926 47817 56958 35659 35847 27785 4 3949 15200 26677 24035 18711 20080 5 4257 7521 14947 20280 12187 14080 Average 9482 34739 49192 42523 37407 27093 SD 6077 24220 3949 6 30478 29812 24408 FIG. 1 shows the transition of the average value of the three groups.

Results As shown in FIG. 1, in the pre-vaccination experiment, the antibody titer in the 0.5 mg group was increased from the day before the second vaccination as compared to the control group. The antibody titer peaks from 6 days to 2 weeks later. Clearly there is an effect of enhancing the action of the vaccine. Even in the 5.0 mg dose group, 0.5
Results similar to those seen in the mg dose group were obtained.

(2) Table 2 shows the effect of mao-bushi-saishin-to on IgG antibody titer after administration of influenza vaccine.
(The numerical values indicate the IgG antibody titer).

[Table 2] Administration after vaccination 1st vaccination 2nd vaccination 2nd vaccination 2nd vaccination 2nd vaccination 2nd vaccination 6 days before day 6 days after 2 weeks after 4 weeks after 8 weeks number of specimens 15 15 13 11 10 10 0.0mg 1 21754 19600 Administration 2 8073 12530 (Control) 3 4814 14562 16488 26270 23789 11472 4 5140 17736 13877 20276 30780 19383 5 5378 27213 37235 36 122 11296 57234 Average 9032 18328 22533 27556 21955 29363 SD 6466 5074 10450 6533 8059 19971 0.5mg 1 13060 18382 22549 Administration 2 6643 15864 22741 17038 27839 7233 3 4049 15497 45479 56970 52776 29563 4 7520 20955 19947 12696 35506 53893 5 1125 12087 11095 Average 6479 16557 24362 28901 38707 30230 SD 3975 2975 11378 19927 10429 19055 5.0mg 1 11021 29688 17243 8810 20582 35488 Administration 2 13285 30724 39708 61522 82625 53704 3 8480 16916 27067 67044 74623 51813 4 5308 15649 52379 78964 82811 45258 5 6763 17075 27754 82019 105701 Average 8971 22010 32830 59672 73268 46566 SD 2876 6718 12095 26519 28313 7122 Further FIG. 2 shows the transition of the average value of the three groups.

Results As shown in FIG. 2, in the post-vaccination administration experiments, the antibody titer peaks 4 weeks after the second vaccination in both the 0.5 mg and 5.0 mg groups. No remarkable effect was observed in the 0.5 mg group, but the antibody titer increased in the 5.0 mg group from 6 days to 8 weeks after the second vaccination as compared with the control group. There is a strengthening effect.

2. Acute toxicity test (LD ₅₀ mg / kg: using a water-based dry extract) Test method Maho-bushi-saishin-to: Mao-bushi-saishin-to used a water-made dry extract prepared in the same manner as described above.

The animal species route of administration LD ₅₀ BDF ₁ mice ♂ oral> 5,000 common symptoms: 2,000mg / kg, but was seen napped phenomenon at 5,000mg / kg administration, the other no abnormal findings, No deaths. Species route LD ₅₀ F344 mice ♂ oral> 5000 F344 mice ♀ oral> 5,000 General symptoms: 5,000 mg / kg hyperreflexia with respect to sounds both sexes in administration, locomotor hyperactivity and salivation and lacrimation It was seen, but no other abnormalities were found and no deaths occurred.

3. Subacute toxicity test F344 rats were treated with 50, 250
It was administered at 1,250 mg / kg / day for 13 weeks. as a result,
In both males and females, the body weight loss and slight changes in blood biochemistry were observed in the 1,250 mg / kg administration group.
In the group administered at 0 mg / kg or less, there was no effect associated with the administration of the drug substance extract bulk powder. No deaths were observed in any of the groups.

[0031]

EXAMPLES Preparation Example Preparation of Mao-bushi-saishin-to extract Mao-bushi, batsushi and spicy, all of the Osaka market products, were minced and mixed at a weight ratio of 4: 1: 3, and mixed in a 10-fold amount. Deionized water was added, and the mixture was refluxed at about 100 ° C. for 1 hour.
After the extract was filtered through a 100-mesh sieve, it was freeze-dried to prepare a mao-bushi-saishin-to extract. The extract obtained at a yield of 15% was used in the following examples.

Example 1 Extract Capsule Mao-Bushi-Saishin-Water Dry Extract 120g Magnesium Aluminate Metasilicate 6g Filler 18g Corn Starch 18g Light Silicic Anhydride 3g Magnesium Stearate 9g Carmellose Calcium 3g Total 177g Therefore, they were mixed uniformly, and 295 mg thereof was filled in a No. 2 hard capsule. In one capsule,
It contains 200mg of dried extract made of Mao-bushi-saishin-sui-yu, and 2 to 9 adults a day is divided into several doses.

Example 2 Fine granules of extract Mao-bushi-saishin-sui-dried water extract 120 g Corn starch 129 g Lactose 335 g Magnesium metasilicate aluminate 8 g Magnesium stearate 8 g Total 600 g According to the above-mentioned formula, mix uniformly, and use a compression molding machine. After compression molding, the mixture was crushed with a crusher and passed through a No. 30 sieve to obtain fine granules. 1 g of this fine granule contains 200 mg of dried extract made of mao-bushi-saishin-sui-water, and 2 to 9 g of an adult is taken several times a day.

Example 3 Extract Granules Mao-Bushi-Saishin-Shoyu Dried Extract 120 g Corn Starch 131 g Lactose 335 g Magnesium metasilicate aluminate 7 g Magnesium stearate 7 g Total 600 g According to the above formula, mix uniformly and compress with a compression molding machine. After molding, the mixture was crushed with a crusher and passed through a No. 12 sieve to obtain granules. 1 g of this fine granule contains 200 mg of dried extract made of mao-bushi-saishin-sui-water, and 2 to 9 g of an adult is taken several times a day.

Example 4 Extracted Tablets Dry extract made of mao-bushi-saishin-shin-water 120 g Carmellose calcium 22 g Carplex 24 g Magnesium aluminate metasilicate 7 g Magnesium stearate 7 g Total 180 g To give a tablet of 180 mg per tablet. One tablet contains 120 mg of dried extract made of mao-bushi-saishin-sui-water, and 3 to 15 tablets of an adult are taken several times a day.

Example 5 Extract soft jelly agent Mao-bushi-saishin-hot water dried extract 120 g Carrageenan 64 g Locust bean gum 18 g Reduced maltose liquid sugar 300 g Xylitol 50 g Apple flavor 3 g Lecithin 40 g Citric acid 5 g Total 600 g According to the above-mentioned recipe After mixing, the mixture was added to 9400 g of heated sterilized purified water with stirring, and after cooling, a soft jelly agent was obtained. 1 g of this soft jelly agent contains 12 mg of dried extract made of mao-bushi-saishin-sui-water, and it is 3 to 1 per day for an adult.
Take 50g in several divided doses.

Example 6 Extract drink agent Mao-bushi-saishin-sui-water dried extract 120 g Reduced maltose liquid sugar 300 g Orange flavor 5 g Ascorbic acid 5 g Lecithin 40 g Polyoxyethylene hydrogenated castor oil 20 g 4-hydroxymethyl benzoate 10 g Total 500 g After uniformly mixing according to the prescription, the mixture was dissolved in heated purified water to make a total volume of 900 mL to obtain a drink. 1 mL of this drink preparation contains 133 mg of dried extract made of mao-bushi-saishin-sui-yu, and it is 30 to 135 per day for an adult.
Take g in several divided doses.

Example 7 Extract Dry Syrup Preparation Dry extract made of mao-bushi-saishin-sui water 120 g Magnesium aluminate metasilicate 29 g Reduced maltose 400 g Grapefruit flavor 6 g β-cyclodextrin 40 g Ascorbic acid 5 g Total 600 g , Summed up. After granulation by an extrusion granulator, the mixture was dried and passed through a No. 12 sieve to obtain a dry syrup. This dry syrup 1
g contains 200 mg of dried extract made of Mao-bushi-saishin-sui-water, and 2 to 9 g of an adult is taken several times a day.

Example 8 Extract Loach Mao-Bushi-Saishin-Water Dry Extract 120g Magnesium Aluminate Metasilicate 30g Polyvinylpyrrolidone 30g Magnesium Stearate 15g Reduced Maltose 618g Peppermint Flavor 12g β-Cyclodextrin 40g Ascorbic Acid 5g Total of 870g According to the prescription, and, and a part of and are uniformly mixed, after compression molding with a compression molding machine,
The mixture was pulverized with a pulverizer, and the remaining amount was added, mixed uniformly, and compressed with a tableting machine to obtain 1.45 g of a troche per tablet. One lozenge contains 200 mg of dried extract made of mao-bushi-saishin-sui-water, and 2 to 9 adults daily are taken in several divided doses.

Effect of dried extract made of mao-bushi-saishin-sui-yu-water Since the immunity of elderly people has generally declined, even when infected with influenza virus, symptoms such as fever are unlikely to appear. Therefore, even if infected with influenza, there are many cases in which death occurs without knowing until pneumonia and the like occur simultaneously. In addition, even if the vaccine is vaccinated, the antibody titer does not increase so much that the effect of the vaccine is hardly recognized. Mao-bushi-saishin-to, when used in combination with a vaccine, activates the weakened immunity and has the effect of increasing the action of the vaccine.

[Brief description of the drawings]

FIG. 1 is a graph showing the effect of mao-bushi-saishin-to on IgG antibody titer before influenza vaccination.

FIG. 2 is a graph showing the effect of Mao-Bushi-Saishin-To on IgG antibody titers after influenza vaccination.

 ──────────────────────────────────────────────────続 き Continuation of front page (72) Inventor Natsumi Higashi 24-1-260 Kita-Ohi-cho, Takatsuki-shi, Osaka

Claims (1)

[Claims] 1. An influenza vaccine action enhancer comprising Mao-Fushi-saishin-to.