JPS6330417A - Anti-allergic agent - Google Patents

Abstract

PURPOSE:To obtain an anti-allergic agent containing a specific compound existing in YAKAN (rhizome of Belameanda chinensis, a plant of Iridaceae family) such as tectorigenin as an active component and effective as a remedy for immediate allergy. CONSTITUTION:The compound of formula (R1 is OH or -O-D-beta-glucose; R2 is methoxy; R1 and R2 may together form methylenedioxy; R3 and R5 are OH or methoxy; R4 is H or methoxy; R6 is H, OH or methoxy) existing in YAKAN is used as an active component of the objective agent. Examples of the compound of formula are tectorigenin (R1, R3 and R5 are OH, R2 is methoxy and R4 and R6 are H), irigenin (R1, R3 and R6 are OH and R2, R4 and R5 are methoxy), etc. The agent is preferably administered usually at a dose of 150-300mg daily as oral drug and 3-60mg daily as parenteral drug for adult.

Description

[Detailed description of the invention] Yakan is a plant of the Iridaceae family, Bela.
mcanda chinensis), and is well known to have antimicrobial and anti-inflammatory effects. In order to develop an anti-allergic agent that is effective for the treatment of immediate-type allergies, the present inventors investigated the effects of various ingredients contained in this hydrangea, and found that the general formula (where R is a hydroxyl group or -D-β-glucose, R2 is a methoxy group, or R and R1 together are a methylenedioxy group, R1 and R6 are a hydroxyl group or a methoxy group, R6 is a hydrogen atom or a methoxy group, R
6 means a hydrogen atom, a hydroxyl group or a methoxy group. ) The present invention was completed by discovering that a compound represented by the formula (hereinafter referred to as a compound of the general formula) has an anti-allergic effect.

The compound of the general formula can be obtained, for example, by the following method. Bela, a plant of the Iridaceae family.
The dried rhizomes of Leman (mcanda chinensis Leman) are extracted using a single or mixed solvent selected from water, methanol, ethanol, ethanol, and acetone by heating from 0°C to a temperature below the boiling point of the solvent used. Alternatively, an extract is obtained by ultrasonic extraction at 0°C to room temperature. This extract can be used as it is, or concentrated, dried, diluted with water, and extracted with ethyl ether.
Partition extraction is performed using benzene, chloroform, methylene chloride, etc. to obtain a partitioned extract.

Further, the distribution residue is subjected to distribution extraction using ethyl acetate, n-butanol, methyl ethyl ketone, etc. to obtain a distribution extract.

The partitioned extract obtained as described above is subjected to column chromatography using a carrier such as silica gel, alumina, reversed-phase silica gel, etc. as it is, or after being concentrated or dried, and the eluate is fractionated. As the elution solvent, water or a single or mixed solvent selected from methanol, ethanol, acetone, acetonitrile, ethyl acetate, chloroform, methylene chloride, benzene, ethyl ether, n-hexane, etc. can be used. The thus obtained fractions can be used as they are, or can be concentrated and dried to obtain fluorescent agent-containing silica gel (manufactured by Merck & Co., Ltd., RP-18F□,
S, etc.) as a carrier of a thin laminate plate, and a developing solvent of water or a single or mixed solvent selected from methanol, acetonitrile, tetrahydrofuran, acetone, ethyl acetate, chloroform, benzene, methylene chloride, ethyl ether, and n-hexane. After being subjected to thin layer chromatography and developed, fractions containing the compound of the general formula identified by ultraviolet (254 nm) irradiation were collected using water or methanol, ethanol, tetrahydrofuran, ethyl acetate, ethyl ether, chloroform. The compound of the general formula can be obtained by recrystallization using one or more mixed solvents selected from , acetone, methylene chloride, benzene, and n-hexane.

A specific example of the method for producing the compound of the general formula is shown below.

Tictrigenin Among the compounds of the above general formula, R3 and R3 are hydroxyl groups,
A compound in which R3 is a methoxy group, R4 and R6 are hydrogen atoms, and R1 is a hydroxyl group is Tectorigenin (Tectorigenin).
genin) and can be obtained as follows.

Specific example 1 Belamcanda chinens
is Leman rhizome) 2.5 kg was crushed, extracted under reflux under heating with 7 volumes of ethanol-water (1:l) for 2 hours, and the extract was heated to remove the solvent under reduced pressure. water 4
9. 428.61 lyophilized extract and lyophilized.
g (yield 16.87%).

This freeze-dried extract was diluted and dissolved with 1.5 portions of water, partitioned and extracted 5 times with 2 portions of ethyl ether, and the partitioned extracts were combined and the solvent was removed under reduced pressure.
．． 20 g were obtained (yield!, 6%).

Furthermore, the distributed water-soluble portion was extracted 5 times with 2 portions of n-butanol, the distributed extracts were combined, the solvent was removed under reduced pressure, and 1 liter of water was added and lyophilized, and the lyophilized extract was 122.
69 g was obtained (yield 4.83%).

Next, 41.20 g of the ethyl ether extract was subjected to column chromatography using 400 g of silica gel (Kieselgel 60.70-230 mesh, manufactured by Merck & Co.), and the mixing ratio of ethyl acetate was sequentially adjusted using a mixed solvent of benzene and ethyl acetate. eluted in increasing amounts. The fraction eluted with a mixed solvent of benzene-ethyl acetate (9:1) was removed under reduced pressure, and was then diluted with n-hexane-ethyl acetate.
Recrystallized with a mixed solvent of tetrahydrofuran (1:1),
2.22 g of slightly yellow needle-like substances were obtained (yield 0.09%).

This pale yellow needle-like substance is described in the literature [Asa Kawase].

Naokazu 0hta, Kazuyoshi Y
Agishita.

Agr, Biol, Chem, 37(1) 1
45 (1973)].

Electron impact mass spectrometry (E
I-MS): 111/Z 300 (M”)
Point: 227-228℃ 4sOH Ultraviolet absorption spectrum λ nm (log ε): mαχ 267 (4,48), 338 (3,52) Nuclear magnetic resonance spectrum (699min d'-DMSO): 12.96 (I
H,s), 8.19 (l H9s).

7.37 (2H, d, J = 8.8 Hz).

6.83 (2H, d, J = 8.8 R2).

6.47 (IH, s), 3.79 (3H, s) + 3
0-Nuclear magnetic resonance spectrum (699min d'-DMSO): 180.2(s
), I 57.1(s).

157.0 (s), 1'53.1 (d).

152.8 (s), 152.4 (s).

131.3 (s), 129.6 (dx2).

121.7 (s), 121.0 (s).

1 14.8 (dX2), l 04.7 (s)
.

93.5(d), 59.5(Q) Irigenin Among the compounds of the above general formula, the compound in which R11R3 and R6 are hydroxyl groups and R1, R4 and R6 are methoxy groups is called irigenin. is,
It can be obtained as follows.

Specific Example 2 The fraction eluted with the mixed solvent of benzene-ethyl acetate (8:2) obtained in Specific Example 1 was subjected to solvent removal under reduced pressure, and the fraction was eluted with the mixed solvent of n-hexane-tetrahydrofuran (l 1). Recrystallization yielded 3.57 g of pale yellow needle-like substances (
Yield 0.14%). This pale yellow needle-like substance is described in the literature [A, 1
~． Ali, N.A., El-emary.

! 11. A, Eshimoghazi, F, M, Darw
ish, A.W., Frahm.

Phytochemistry, 22(9) 206
1 (1983), the physical and chemical properties of irigenin.

Electron impact mass spectrometry (E
I-MS): m/z 360 (M") Melting point: 180-183 °C Ultraviolet absorption spectrum A; Hr nm (log ε): 269 (4,47
), 336(3,59)B)--1° infrared absorption spectrum rnQ engineering α.

3440.1662. l 624.1580゜l 51
4 Proton nuclear magnetic resonance spectrum (δl) [)Ill in d'-DMSO): 12.
93 (IH, s), 8.26 (IH, s).

6.68 (l H, d, J = 2 Hz).

6. 7 3 (l H, d, J =
2) (Z).

6.48 (IH, s), 3.81 (3H1s).

3.79 (3H, s), 3.75 (3H, 5) 13
0-Nuclear magnetic resonance spectrum (699m in d8-DMSO): 179.9(s
), l 57.0(s).

153.9(d), 152.8(s).

152.5 (s), 152.3 (s).

1 49.8 (s), l 36', 7 (s
).

1 3 1.4 (s), 1 25.6 (s).

1 2 1.6(s), l l O,3(d
).

105.1(d), 104.7(s).

93.6(d), 59.5(qx2), 5
5.8(q) (blank below) Irisflorentin Among the compounds of the above general formula, the compound in which R1 and R7 together are a methylenedioxy group, R3, R, R6 and R6 are a methoxy group is irisflorentine. Chin (Irisr.
lorentin) and can be obtained as follows.

Specific Example 3 The fraction eluted with the mixed solvent of benzene-ethyl acetate (7:3) obtained in Specific Example 1 was removed under reduced pressure and recrystallized with n-hexane-ethyl acetate (1:1). 1.77 g of colorless fine needle-like material was obtained (yield 0.07%).
). This colorless fine needle-like substance is described in the literature [Naokata M
orita, Munehisa ArAr15a.

Yoshikazu Kondo, Tsunemat
Su Takemoto, Chem.

Pharm, Bull, 21(3) 600(1
The physicochemical properties of irisflorentin were consistent with those described in [973)].

Electron impact mass spectrometry (E
I-MS): n+/z 386 (M”)
Point: 167-168℃ Ultraviolet absorption spectrum λ g” nm (log
e): 266 (4, 41), 324 (3, 6
8) Infrared absorption spectrum is coming! ! -':1664
, l 630, 1 582.1 5 10 elemental analysis
Theoretical value C62,17H4,70 Measured value C62,3
6H4,61 Proton nuclear magnetic resonance spectrum (δppm in d'-DMSO): 8.19 (l
H,s), 6.88 (I H,s).

6.85 (2H, br, s), 6.14 (2H, s
).

3.93 (3H, s), 3.81 (6H, s).

3.73 (3H,5) R30-nuclear magnetic resonance spectrum (δppm in d''-DMSO): 173.3 (s
), l 53.5(s).

152.2 (sx3), 151.2 (d).

140.4 (s), 137.9 (s).

135.5 (s), 127.0 (s).

123.8(s), 113. Q(3).

107.3 (dx2), 102.1 (t).

92.9(d), 60.2(q), 59.
6(q).

55.9 (qx2) Iridine Among the compounds of the above general formula, the compound in which RI is -0-D-β-glucose, R8, R4 and R6 are methoxy groups, and R1 and R8 are hydroxyl groups is Iridine.
) and can be obtained as follows.

Specific example 4 n-butanol dry extract obtained in specific example 1 122.69
g to silica gel (Kieselgel 60, 70-
The mixture was subjected to column chromatography using 700 g of 230 mesh (manufactured by Merck & Co., Ltd.), and eluted with a mixed solvent of chloroform and methanol while increasing the methanol mixing ratio. The solvent of the fraction eluted with 5 volumes of a mixed solvent of chloroform-methanol (9:I) was removed under reduced pressure, and recrystallized with methanol-water (5:I) to obtain 900 g of pale yellow needle-like material (harvested rate 0.36%). This pale yellow needle-like substance is described in the literature [A, A, Ali, N, A, El-em
ary, M.A., El-moghazi.

F, M, Darwish, A, Buy, Frahm,
Phytochemistry.

22(9) 2061 (1983)].

Tweel Dobutsorption Mass Spectrometry (F
D-MS): m/z 523 (MH+) Melting point:
205-207℃ H30H Ultraviolet absorption spectrum λ nm (log ε): -256 (4,80), 320 (sh, 3.88)
1 in the infrared absorption spectrum ν. s −/・spit χ
3396, +658.1622, 1588° Proton nuclear magnetic resonance spectrum (δppm in d'-DMSO): 12.81 (I
H, s), 8.92 (IH, brs).

8.38 (I H, s), 6.87 (I H, s)
.

6.75 (IH, d, J=1.95H2).

6.70 (IH, d, J = 1.95 R2).

5.08 (tH, a, r = 9.0 Hz).

3.81 (6H, s), 3.74 (3H, s).

3.0-3.7(m) I30-Nuclear magnetic resonance vector (δpi) min d@-DMSO): 180.2(
s), 156.2(s).

154.5(d), 152.6(s2).

151.9 (s), 149.8 (s).

136.7 (s), 132.7 (s).

125.4 (s), I 21.8 (s).

110.3(d), lO6,4(s).

105.1(d), 100.4(d).

94.0(d), 77.0(d), 76.5(
d).

73.0(d), 69.7(d), 60.7(
t).

59.9(q), 59.6(q), 55.8(
q) Tectlizine Among the compounds of the above general formula, a compound in which R1 is -0-D-β-glucose, R7 is a methoxy group, R3 is a hydroxyl group, R4 and Ro are a hydrogen atom, and R1 is a hydroxyl group is called tectolizine. It can be obtained as follows.

Specific Example 5 The fraction eluted with the mixed solvent of chloroform-methanol (8:2) obtained in Specific Example 4 was removed from the solvent under reduced pressure, and recrystallized with water-methanol (1:1) to obtain colorless needles. 14.67 g of a similar substance was obtained (yield 0.59%). This colorless acicular substance has been described in the literature [Asa Kawase, Nao
Kazu 0hta, Kazuyoshi Yagis
hita, Agr, Biol, Chem,, 3
7 (1) 45 (1973)] described in Tectorizin (T
ectoridin).

Tweel Dobutsorption Mass Spectrometry (F
D-MS): m/z 485 (M+Na'').

462 (M") Melting point; 262-265°C Ultraviolet absorption spectrum λ': Xg" nm (log ε).

268 (4,54), 333 (3,53) infrared absorption spectrum ν to %2, -/.

3368.1658, 1 6 14, 1 5B2° Proton nuclear magnetic resonance spectrum (δppm in d@-DMSO): 12.82
(LH, br, s), 9.3 (lH, br, s).

8.3 1 (IH, s).

7.3 9 (2H, d, J = 8.5 Hz).

6.8 5 (IH, s).

6.8 3 (2H, d, J = 8.3 Hz).

5.06 (IH, d, J = 7.2 Hz).

3.8 1 (3H, s), 3.0-3.8 (m)
130-nuclear magnetic resonance spectrum (δppm in d''-DMSO): 180.9 (s
), 157.6(s).

156.6(s), 154.2(d).

153.0 (S), 152.4 (S).

133.0 (s), 130.0 (d2).

122.4 (s), I 21.2 (s).

115.2 (dX2), 106.8 (s).

100.8(d), 94.4(d).

77°5 (d), 76.9 (d), 73.
4(d).

70.1(d), 61.1(t), 60°3(
q) Next, the fact that the compound of the general formula, which is the active ingredient of the antiallergic agent of the present invention, has an antiallergic effect will be explained using experimental examples.

Experimental Example <PCA reaction inhibitory effect> Wistar male rats, whose hair had been shaved the previous day, were sensitized by intradermally administering 0.1-mg of anti-ovalbumin IgE serum, and 48 hours later, 0.1-ml anti-ovalbumin IgE serum was administered intradermally to male Wistar rats whose hair had been shaved the previous day.
5% Evans blue solution! - by intravenously administering P
CA (passive cutaneousan)
aphy laX iS passive cutaneous anaphylaxis) reaction was elicited. Thirty minutes later, the mice were sacrificed by exsanguination, and after the skin was dissolved, the amount of dye leaked was determined by colorimetry. Further, the compound of the general formula was intraperitoneally administered 30 minutes before the reaction induction, and the inhibition rate was calculated using the following formula.

Amount of leaked dye when compound of general formula A is not administered Amount of dye leaked when administering compound of general formula B The results are shown in Table 1.

Table 1 From the results, the compound of the general formula suppresses the PCA reaction,
It was found to be effective in treating allergies.

Next, an acute toxicity test of the compound of the general formula was conducted using ddY mice (each dose, IIRI 0 mice), and there was no death in any case at oral doses up to 2000 mg/kg.

Furthermore, the compounds of the general formula can be administered to animals and humans as such or together with conventional pharmaceutical carriers. The dosage form is not particularly limited and may be selected and used as appropriate, including oral preparations such as liquids, powders, granules, tablets, enteric-coated preparations and capsules, and parenteral preparations such as injections and suppositories. Can be mentioned.

In order to achieve the desired effect as an oral agent, the usual adult dosage is 150 to 300 mg of the compound of the general formula, divided into three doses per day, depending on the age, weight, and severity of the disease of the patient. It seems appropriate to take it.

In the present invention, oral preparations such as tablets, capsules, and granules are manufactured according to conventional methods. The tablet contains a compound of the general formula gelatin, starch, lactose, magnesium stearate,
Capsules are manufactured by mixing with pharmaceutical excipients such as talc and gum arabic and excipients.Capsules are prepared by mixing the compound of the general formula with an inert pharmaceutical filler or diluent, and forming hard gelatin capsules. Manufactured by filling soft gelatin capsules, etc. Syrups and elixirs are
The compounds of the present invention can be used as sweeteners such as sucrose, preservatives such as methylparaben and propylparabens, colorants, seasonings,
Manufactured by mixing with fragrances and adjuvants.

In order to exert the desired effect as a parenteral agent, although it varies depending on the patient's age, weight, and degree of disease, it is usually necessary for adults to intravenously inject 3 to 60 mg of the compound of the general formula per day. Subcutaneous and intramuscular injections are considered appropriate.

This parenteral preparation is manufactured according to a conventional method, and distilled water for injection, physiological saline, aqueous dextrose solution, vegetable oil for injection, propylene glycol, polyethylene glycol, etc. can generally be used as a diluent. Furthermore, if necessary, a bactericidal agent, an isotonizing agent, a stabilizer, a preservative, a soothing agent, etc. may be added as appropriate. In addition, from the viewpoint of stability, this parenteral preparation can also be filled into ampoules and the like, frozen, water removed by ordinary freeze-drying techniques, and a liquid preparation prepared from the freeze-dried product immediately before use.

Other parenteral preparations include liquid preparations for external use, liniments such as ointments, suppositories for rectal administration, and the like, which are manufactured according to conventional methods.

Next, the present invention will be explained in more detail with reference to examples, but the present invention is not limited thereto.

Example 1 2.5 g of tictrigenin obtained in Example 1 was made into a fine powder, mixed with 90.5 g of lactose and 7 g of magnesium stearate, and the mixture was compressed into tablets with a diameter of 20 mm using a single-shot slug tablet machine. A slug tablet weighing 2.3 g was prepared, crushed with an oscillator, sized, and sieved to obtain good granules with particles of 20 to 50 mesh.

Each gram of this granule contains 25 mg of tictrigenin, and the dosage is 2 to 4 g three times a day, depending on the symptoms.

Example 2 2.5 g of irigenin obtained in the above specific example 2 was made into fine powder, and this was mixed with 905 g of lactose and 7 g of magnesium stearate.
The mixture was then compressed using a single-shot slug tablet machine to make slug tablets with a diameter of 20 IIllT + 1 weight N 2.3 g, which were crushed using an onlator, sized, and sieved to form 20 to 50 mesh tablets. Granules with good particles were obtained.

This granule contains 25 mg of irigenin in 1 g, but depending on the symptoms, take 2 to 4 g at a time, three times a day.

Example 3 70 g of microcrystalline cellulose and 5 g of magnesium stearate were added to 25 g of iris florentin obtained in Example 3 above.
was added and mixed, and this mixture was compressed using a single-shot tablet machine to produce tablets with a diameter of 9 mm and a weight of 1200 mg.

Each tablet contains 50 mg of irisflorentin, and the dosage is 1 to 2 tablets, 3 times a day, depending on the symptoms.

Example 4 70 g of microcrystalline cellulose and 5 g of magnesium stearate were added to 25 g of irisine obtained in Example 4 and mixed, and the mixture was compressed into tablets with a single-shot tablet machine to form tablets with a diameter of 9 m.
Tablets with a weight of 200 mg were produced.

Each tablet contains 50 mg of irisine, and depending on the symptoms, take 1 to 2 tablets at a time, three times a day.

Example 5 The irigenin log obtained in Example 2 was made into a fine powder, mixed with 90 g of lactose, and 500 mg each of the powder was filled into hard capsules to obtain capsules.

This capsule contains 50 mg of irigenin per capsule, and the dose is 1 to 2 capsules taken 3 times a day depending on the symptoms.

Example 6 10g of iris florentin obtained in Example 3 was made into a fine powder, mixed with 95 g of lactose, and 500 mg each of the powder was filled into hard capsules to obtain capsules.

This capsule contains 5 irisflorentin in each capsule.
It contains 0mg, but depending on your symptoms, take 1 to 2 capsules at a time, 3 times a day.

Example 7 50 g of tictrigenin obtained in Example 1 was added to 1 volume of distilled water for injection heated to 60°C according to the usual method for injections, turbid, made isotonic with sodium chloride, and then sealed in an ampoule. .

This injection contains 50 mg of tictrigenin in the IJ. This injection is administered intravenously or intramuscularly in 2 to 4 doses at a time, depending on the symptoms.

Example 8 Distilled water for injection prepared by heating 50 g of iristhrofluorentin obtained in Example 3 to 60°C according to the usual method for injections! In addition, the mixture was suspended, made isotonic with sodium chloride, and then sealed in ampoules.

This injection contains irithrofluorentin 50B in l-. This injection is administered intravenously or intramuscularly at a dose of 2 to 4 doses depending on the symptoms.

Claims (6)

[Claims] (1) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (However, R_1 is a hydroxyl group or -O-D-β-glucose, R_2 is a methoxy group, or R_1 and R_2
together represent a methylenedioxy group, R_3 and R_
5 means a hydroxyl group or a methoxy group, R_4 means a hydrogen atom or a methoxy group, and R_6 means a hydrogen atom, a hydroxyl group or a methoxy group. ) An anti-allergic agent containing the compound represented by as an active ingredient. (2) The antiallergic agent according to claim 1, wherein in the above general formula, R_1 and R_3 are a hydroxyl group, R_2 is a methoxy group, R_4 and R_6 are a hydrogen atom, and R_5 is a hydroxyl group. (3) The anti-allergic agent according to claim 1, wherein in the above general formula, R_1 and R_2 together represent a methylenedioxy group, and R_3, R_4, R_5 and R_6 represent a methoxy group. (4) In the above general formula, R_1, R_3 and R_
The anti-allergic agent according to claim 1, wherein 6 is a hydroxyl group, and R_2, R_4 and R_5 are methoxy groups. (5) In the above general formula, R_1 is -O-D-β-glucose, R_2, R_4 and R_5 are methoxy groups,
The antiallergic agent according to claim 1, wherein R_3 and R_6 are hydroxyl groups. (6) In the above general formula, R_1 is -O-D-β-glucose, R_2 is a methoxy group, R_3 is a hydroxyl group, R_
4 and R_6 are hydrogen atoms, and R_5 is a hydroxyl group, the antiallergic agent according to claim 1.