JPS5852273A - Aconitine-alkaloid, its preparation, and antiphlogistic anodyne containing it - Google Patents

Abstract

NEW MATERIAL:An aconitine-alkaloid shown by the formulaI(R1 is lower alkyl; R2 is H or OH; R3 is residue of linoleic acid, palmitic acid, stearic acid, or linolenic acid; B2 is benzoyl). EXAMPLE:A compound shown by the formula II. USE:An antiphlogistic anodyne. Showing improved suppressing action on carragenin edema and analgesic action. PROCESS:An alcohol extracted solution of bulb of Ranunculaceae Chinese aconite is separated by column chromatography, to give aconitine-alkaloid shown by the formulaI.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel aconidial alkaloid, a method for producing the same, and an anti-inflammatory analgesic containing the same.

Tsushi is the bulb of atricapt and its congenerous plants belonging to the family Ceramicaceae, and has been used as a treatment for neuralgia and rheumatism in traditional Chinese medicine since ancient times.

The true identity of this medicinal ingredient is believed to be the alkaloid component contained in tsuji, and aconidial alkaloids such as a]nithine, mesaconitine, hibaconitine□, and jusaaconitine have been isolated and studied in detail. However, these aconidial alkaloids are extremely toxic, as they have been used in poisonous arrows since ancient times, and the effective dose used clinically for the above-mentioned diseases is very close to the toxic dose. It has been extremely difficult to use Tsushi for these diseases because the amount of poison and the amount of toxicity varies depending on each individual and their symptoms and conditions.

As a result of repeated research on the anti-inflammatory and analgesic components of 1 g of tsuji, which is obtained by re-processing tsuji and raw shiwawa tsuji, the present inventors succeeded in isolating a new aconidial alkaloid that is less toxic and more effective than acoethyl. The present invention has now been completed.

The novel aconidial alkaloid of the present invention has the following general formula (
I).

(In the formula, R represents a lower alkyl group, R2 represents a hydrogen atom or a hydroxyl group, R3 represents each acid residue of linoleic acid, balmitic acid, oleic acid, stearic acid, or lylisic acid, and fjz represents a penlyl group.) General formula ( Preferred examples of the lower alkyl group for I) R□ include a methyl group, an ethyl group, and the like.

Preferred examples of the compounds of the present invention included in the above general formula (I) include compounds represented by the following formulas (n) to (■).

ocn3utn3 (R3 and BZFI in each formula have the same meanings as above.

) The aconidial alkaloid of the present invention can be obtained by separating an alcoholic extract of 100% of 100% of the total amount of alcoholic acid using a column method.

It is possible to use any of the known products obtained by reconditioning and reprocessing raw Tsuji used as raw materials, such as those obtained by soaking raw Tsuji in bittern liquid, boiling, soaking in sugar solution, and steaming and drying. The above-mentioned alcoholic extract of Hirofuji can be obtained by extracting the Hirofuji with, for example, methanol, and then partitioning and extracting with 00 form-methanol-water. The method of the present invention involves passing the above-mentioned extract through a conventional column, such as a column packed with silica gel, and using an appropriate eluent such as 00 form-ethyl acetate or 00 form-ethyl acetate-methanol-water. Elution is carried out by passing the eluate through a column filled with, for example, alumina, and eluting it with an eluent of beshizeshi or penzeshiacetoshihiro.

The details and various physical and chemical properties of the compound of the present invention obtained by the above method are as shown in the Examples below.

The compound of the present invention thus obtained has anti-inflammatory analgesic activity, low toxicity, and is useful as an anti-inflammatory analgesic.

The present invention also provides such anti-inflammatory analgesics.

The anti-inflammatory and analgesic agent of the present invention contains an aconidial alkaloid represented by the above general formula (I) as an active ingredient, and is prepared into a suitable pharmaceutical composition according to a conventional method using an arbitrary pharmaceutical carrier or excipient. prepared and ready for use. In view of the above, various commonly used pharmaceutical carriers and excipients can be used depending on the form of the pharmaceutical composition to be obtained. For example, in order to obtain tablets for oral administration, 55 cell granules, powders, etc., excipients commonly used in the art, such as calcium carbonate, calcium ricinate, decide, Sucrose, lactose, talc, magnesium stearicate, ze5tish, polyvinylpyrrolidoshi, arabicum,
Sorbit, microcrystalline cellulose, polyethylene diglycol, carbo+dimethyl cellulose, silica, polyvinyl acetal, diethyl chloride! Noacetate, methylcellulose, shellac, etc. can be used.

Tablets may be coated by methods well known in the art.

In addition, liquid preparations for oral administration, specifically aqueous or oil-based suspensions, solutions, drops, intravenous pills, etc., and injections for parenteral administration, specifically aqueous or oil-based Suspensions, solutions, powder fillers that dissolve before use, lyophilizers, etc. are prepared by conventional methods. When preparing the form of a second solution, diluents commonly used in the art such as distilled water, ethyl alcohol, propylene glycol, isostearyl ethyl ocyanide, polyol + isostearyl sulfide alcohol, etc. are used. Alcohol, etc. can be used.

Injectable preparations are sterilized and are isotonic with blood, so salt, glucose, etc. may be added to make them an isodominant solution.

Furthermore, the pharmaceutical composition may be prepared in the form of a suppository for rectal administration or in the form of an external preparation such as an ointment, cream, poultice, etc. for topical administration. Preparation into a 2-isomorphic form can be carried out according to a conventional method, and a conventional carrier can be used. For example, examples of carriers that can be used to prepare the form of suppositories include polyethylene cyclohexylcol, 5-nolysiloxane, cacao butter, gelatin, fatty acid chloride, and the like. It is administered by various routes depending on the form. The dose, of course, depends on the route of administration (
(Dosage form) can be determined as appropriate depending on the patient's medical condition, age, etc., but usually the dosage per adult per day in terms of active ingredient compound is approximately .
The dosage may range from 0.5 to 100011 F, preferably from about 10 to 500 capes, and may be administered in 1 to 3 doses.

Examples of the production of the compounds of the present invention are listed below as examples.

Example 1 25 tlk of methanol was added to 18.411 of the powder of 1 child, left to wrinkle at room temperature for 24 hours, and then heated. Add methanol 251 to the residue and operate in the same manner. This operation is repeated 10 times, the ν liquid is collected, and the solvent is distilled off under reduced pressure at 60° C. or lower to obtain methanol extraction product 850f as a residue. This methanol extraction process is performed using 00 form-methanol-
Partition extraction was carried out with water (5:6:4), the lower form-methanol layer was separated, the solvent was distilled off under reduced pressure, and the resulting residue was suspended in 500 d of dihydrochloric acid aqueous solution. Extract with lum 2t. Is the 00 form layer ``nat bicarbonate''?
After washing with an aqueous solution of chloroform and then with water, the solvent was distilled off under reduced pressure to obtain chloroform extraction process 1332 as a residue.

The obtained OO form extractor 13o2 was dissolved in ethyl acetate 200R1, sprinkled on silica gel (60-230 mesh, manufactured by Merck & Co., Ltd.) + 5Of, the solvent was distilled off under reduced pressure, and the mixture was added to a silica gel column (silicage Jl/: 1. 5K
R160-230 mesh (manufactured by Merck & Co., Ltd.) was layered with 00 form-ethyl acetate (1:l), 900 form-acetic acid 1 pre-methanol (5:6:l), and then OQ form-ethyl acetate-methanol. - Elute sequentially with water (5:6:l:1). Next, the OO form-ethyl acetate-methanol-water elution fraction +7f was added to the alumina powder [neutral aluminum oxide, activity II].
(ALrnina Woml Associate N, Akt ] [[)
1, 7 kg, 1 F'oa
Elute sequentially with a solvent system of 0:1.50:1.40; 1.30:1, 20:1 and 10:1). Each eluent was divided into two parts based on the thickness of the thin layer, and the solvent was distilled off under reduced pressure.

Thus, the compound 600q of the formula (II), the compound 630■ of the formula @), the compound 62011F of the formula (to), and the compound 62011F of the formula (
Compound 48011F of V) is obtained.

The physical property values and analysis results of each compound of the present invention obtained are shown.

■ Compound of formula <TI) 1) (a) J): +12.4 (C-1,94, Rio
2) Infrared absorption spectrum (RI00 form) analysis 2 3503 (Ar), 2925.1717.1282
, -1096 cm shows the absorption maximum.

3) Ultraviolet absorption spectrum (ethanol) analysis: 227%
[0=16800] indicates the maximum of K absorption.

4) 1H nuclear magnetic resonance spectrum (cnct3) analysis: The following signal (JPPIII) is shown.

H) 5) 13C nuclear magnetic resonance spectrum (cnct3) analysis: The following shiftal is shown.

'174.8 (C of carbonyl derived from fatty acid
) 78.9 (C at positions 14 and 15) 6) FD quality mass spectrum 1 analysis N/Z-851 (molecular ion of a compound in which R3 is an oleic acid residue), Tomi/Z-849 (R3 is a linoleic acid residue) The molecular ion of a compound in which R3 is an acid residue) shows a peak at m/Z=825 (the molecular ion of a compound in which R3 is a pallic acid residue).

7) Properties and solubility: - Colorless, odorless oily substance, ether, lyoform,
Soluble in benzene, ethanol, methanol, acetone, ethyl acetate, pyridyl, dimethyl sulfonate, and insoluble in water.

8) Thin layer 0 Matoku 5 Fi 1 analysis Nijiri Kagel 6QF25. (pre-coated,
Q, 25 Proverbs, manufactured by Merck & Co., Ltd.) in a thin layer with a strong acidity water saturated ether: Hechusashi (
3:l) shows an Rf value of 0.5.

9) When hydrolyzed with 10% sodium chloride and dry methanol, dioxycyanine, fatty acid methyl [methyl linoleate (615G), methyl balmitate (1
9,5%), methyl oleate (19,5%), methyl stearate (image weight), methyl linoleate (fine f)) and methyl benzoate.

10) Geo+Sun-Water (1:1) 120°, 4
When heated for 0 minutes, besylyyldeo+cyaconine and fatty acids (linoleic acid, valmitic acid, oleic acid, stearic acid, lylisic acid) are obtained.

■ Compound of formula @) 1) [α], +6.0 (C=5.14, ri00
2) Infrared absorption spectrum (RI00Nelm) analysis: 3946 (Ay, ), 2922.1721.1277
The maximum absorption is shown at 1097α.

3) Ultraviolet absorption spectrum (ethanol) analysis: 227a
The absorption maximum is shown at m, (g=21000).

4) 1H nuclear magnetic resonance spectrum (cnct3.) analysis:
Indicates the next shifnal.

(H at position 14) s,) 13C nuclear magnetic resonance spectrum (CDC13)
Analysis: The following shifnal (J) is shown.

174.7 (C of carbonyl derived from fatty acid)
165.6.132.9,”29.9.129.6.1
28.3 (C derived from besilyyl group) C) 91.4.89.8.83.2.82.1.75.7.
73.9.70.4 (8.16, summer, 6.18.
13.3rd C) 78.7 (14, I5 C)
) 6) Fl) Mass spectrum analysis: m/Z=867 (molecular ion of a compound in which R3 is an oleic acid residue), no/Z-865 (molecular ion of a compound in which R3 is a linoleic acid residue) ,ML/Z=841C
The molecular ion of a compound in which R is a valmitic acid residue) shows a peak.

7) Properties and solubility: Colorless and odorless oily substance, containing ether, ethyl acetate, tetrachloride, carbon tetrachloride, benzene, ethanol, methanol, acetone, pyri! It is soluble in dimethyl sulfate and insoluble in water.

8) Thin layer 0 matoku 5 fi 1 analysis Nijirica gel 60F254 (Pra-coatgtL,
Thin layer 0 matoku 5 using 0.25 ink (manufactured by Merck & Co.)
When developed with strong asimonia water saturated ether, it shows an Rf value of 0.5.

9) Hydrolysis with 10% sodium methocide and dry methanol yields a small amount of fatty acid methyl (methyl linoleate (64%), methyl baltrinate (20%), methyl oleate (169G), methyl stearicate (
(trace amount), methyl linoleate (trace amount)] and methyl benzoate are obtained.

10) Dio + Sun - Water (1:I) at 120℃, 4
When heated for 0 minutes, besiriylaconine and fatty acids (linoleic acid, valmitic acid, oleic acid, stearic acid,
riluic acid) is obtained.

■ Compound 1) (α]ka, + 13.5 (C= 4.3
3, Ri00form) 2) Infrared absorption spectrum (Ri00*
Lum) Analysis 2 3511 (Jr,), 2929.1'
The absorption maximum of 718.1094cHI-1 is shown.

3) Ultraviolet absorption spectrum (ethanol) analysis: 227n
The maximum absorption is shown at vm (a-17000).

4) 1H Nuclear Magnetic Resonance Spectrum (CDC1,J) Analysis: The following signal is shown.

H) (H at position 6) 2.33 (3H#) (-N-CH,OH)5
) C nuclear magnetic resonance spectrum (chct3) analysis:
The following signal (Jlc) is shown.

174.7 (C of carbonyl derived from fatty acid)9
1.6.90.1.85.0.83.2.80.0.7
4.0 (C at positions 8, 16, 1, 6, 18, 13) 78.
9 (C at positions 14 and 15) 6) FD mass spectrometry analysis: wt/Z-837 (0 molecular ion of a compound where R is an oleic acid residue), m/Z = 835 (R3 is a linoleic acid residue) molecular ion of the compound that is the group), +a/Z=
811 CR, is the molecular ion of the compound in which valmitic acid residue is present).

7) Properties and solubility: A colorless, odorless oily substance that can be dissolved in nitrogen, ethyl acetate, carbon tetrachloride, ethanol, tetrahydrofuran, methanol, acetate, pyridine, and dimethylsulfate. It is soluble and insoluble in water.

8) Thin layer 0 mad j rough 1 analysis:　　.

Silica gel 60F25. (Pry-coated
, 0.25, manufactured by Merck & Co., Ltd.), and shows an Rf value of 0-5 developed with strong ammonia water saturated ether:Hejisashi (5:1).

9) 10tinodium meth + side - When hydrolyzed with dry methanol, Shibakonishi, fatty acid methyl [methyl linoleate (58), methyl paltrisitate (23),
Methyl oleate (19T), methyl stearate (I
I-1), methyl linoleate (trace amount)] and methyl benzoate are obtained in equal amounts.

■ Compound of formula (V) 1) [a]i: + 13.8 <c-+, 26
, Ri00*lum) 2) Infrared absorption spectrum (Ri00*lum) analysis: 3495(7r,), 2924.1717.
It shows an absorption maximum at 1280.1099clII.

3) Ultraviolet absorption spectrum (ethanol) analysis = 227
The maximum absorption is shown at nor (#=18700).

4) 1H nuclear magnetic resonance spectrum (chct, ) analysis:
The following signal (Jppv*) is shown.

2.31 (3H' I) (>A' -C
H30H) 5) 13C nuclear magnetic resonance spectrum (cD
ct3) Analysis: Shows the following signal (δC).

174.9 (fatty acid wild vegetable carbonyl C)
165.8.133.2.129.7.129.4.1
28.4 (C derived from benlyyl group) 78.8 (C at positions 14 and 15) 6) FD mass spectrometry analysis: m/Z = 853 (molecular ion of a compound where Rs is an oleic acid residue), III/ Z=851 (molecular ion of a compound in which R3 is a linoleic acid residue), w, /Z=82
7 (molecular ion of a compound in which R is a valmitic acid residue) shows a peak.

7) Properties and solubility: A colorless, odorless oily substance that contains ether, diform, ethyl acetate, tetrachloride, carbon tetrachloride, benzene, ethanol, methanol, acetate, pyridine, dimethyl sulfate. It is soluble in raw meat and insoluble in water.

8) Thin layer 0 mattoku 5 fi 1 analysis Nijirica gel 6Q F254 (Pre-coated
, 0.25 U, manufactured by Merck & Co., Ltd.) in a thin layer O-magnetic acid graphite with strong ammonia water saturated ether, it shows an Rf value of 0.4.

9) SOS sodium meth + side - Hydrolysis with dry methanol produces mesaconine, fatty acid methyl [methyl linoleate (57%), methyl valmiticate (32%)
, methyl oleate (11), methyl stearate (
1! (amount), methyl lylisinate (trace amount)] and methyl benzoate are obtained in equal amounts.

Song 10) Geo+Sun-Water (g:I) -t'120℃
, heating for 40 minutes yields beslyyl mesaconic acid and fatty acids (linoleic acid, paltritic acid, oleic acid, stearic acid, liluic acid).

Next, examples of formulations of the present invention will be shown.

Formulation Example I Compound of formula av) 50wIg Lactose 650!
280 l
Iv Human 0+Siv 0 Pill Cell 0-S 20q
10,100 O per 1 pack Granules are prepared using the above blending.

Formulation example 2 Compound of formula (V) 0.5 q milk
Sugar 90.5 W crystal t
Ruth 0-su 54sv Shido + Shizu 0 Pill Starch 30 Misaki Taru Ri
4
q Magnesium stearate 1 drop per 1 tablet 180q Am tablets at the above mixing ratio.

Formulation example 3 Compound of formula (Y)　　　　　10℃Product name) 1 piece 1000 Misaki Prepare full opening with the above mixing ratio.

Formulation Example 4 Compound of Formula (II) 1. Of white petrolatum 24. Of stearyl alcohol 22.0 f 0 pyrene glycol 12° Of 5 sodium uryl sulfate 1.5 f ethyl benzoate 0.0025 t Palau + 0 pill 0.0015 F purified water
Appropriate amount Total amount: 100 f Prepare an ointment with the above mixing ratio.

Next, experimental examples will be shown regarding the acute toxicity and pharmacological effects of the compounds of the present invention and aconitine and mesaconitine as controls.

Experimental Example 1 (Acute Toxicity) The test drug was subcutaneously administered to 0ddy male mice weighing around 22f, and LD5o was determined from the number of deaths 1 hour after administration. The results are shown in Section 1H below.

Table 1 The same tests were carried out on the compounds of formulas (2) to (2) of the present invention, and as a result, all of them showed the same LD5 value as the compound of formula (v) above.

The above results show that the compound of the present invention is 20 to 200 times less toxic than the control aconitine and mesaconitine.

Experimental Example 2 (Strength 5 Genishi Edema Suppression Effect) After measuring the volume of the right leg of male Wistar (t'zz tαr) rats weighing around 130, the test drug was immediately administered. 0.5
After an hour, 0.1 d of charazanidi (1%) as an inflammatory agent was injected subcutaneously into the right heel to induce swelling, and the volume of the paw was measured 2 and 3 hours later.

The edema rate (%) was determined as follows from the constant volume (CVt) 2 or 3 hours after the injection on the inflammation side and the footfall volume (Fa) before the injection on the inflammation side.

Edema rate (*) = -X 100 r road The results are shown in Table 2 below.

Experimental Example 3 (Analgesic effect) The test drug was subcutaneously administered to dcLy male mice weighing around 22f, and 30 minutes later, the test drug was 0.61! l'l-acid is Q, l
ml/10 was administered intraperitoneally, and after 0 minutes, 1
The number of times of writing (W'ritki) for 0 minutes was measured.゛The results are shown in Table 2 below.

Table 2 The same test was conducted on the compounds of formulas (1) to (5) of the present invention, and as a result, they showed approximately the same edema rate (-) and analgesic effect.

From the results in Section 1, it is clear that the compounds of the present invention have excellent edema-suppressing effects and analgesic effects. This 21st again! l! Considering the results shown in Table 1 and the results shown in Table 1 above, it can be seen that the compound of the present invention can be used as a significantly superior analgesic drug compared to aconitine and mesaconitine as controls.

(Above) Procedural amendment (own ship 1, indication of the case Patent Application No. 150134 of 1982 3, person making the amendment Relationship with the case Patent applicant Taiho Pharmaceutical Co., Ltd. (and 1 other person) 4 representatives Person 5 Date of amendment order Issue 6. Number of inventions increased by amendment Contents of amendment 1 The statement in the "Claims" section of the specification is corrected as shown in the two papers.

2 "General formula (I)" stated on page 4 of Specification 1iF is corrected as per Geto.

" 3. "Formula (I)" to "Formula 1 (V)" described on pages 5 and 6 of the specification are respectively corrected as follows.

” 4 On page 12, line 6 of the specification, [Almima Wotl
m J Toaru wo 1-Alumima Wazlm
Correct it to J.

5. On the bottom line of page 27 of the specification, the phrase “1 hour” was replaced with “24
"Time," he corrected.

6 The phrase “intraperitoneal administration” in line 12 of specification 29N is replaced with “
Corrected to ``intraperitoneal administration.''

7 In line 12-13 of specification No. 29ji, the phrase “Writkisu” is replaced by “Writkisu”.
kigy ) Correct it as J.

(The above) ■ General formula (in the formula, R□ is a halogen group, R2 is a hydrogen atom or a hydroxyl group, R3 is each acid residue of linoleic acid, valmitic acid, oleic acid, stearic acid, or liluic acid, and B1 indicates a penlyyl group.) Aconine alkaloid represented by.

■ The alcoholic extract of rotsutsuji was separated by column chromatography to obtain the general formula (in the formula, R is lower alkyl, R2 is a hydrogen atom or a hydroxyl group, R3 is linoleic acid, balmitic acid, oleic acid,
Each acid residue of stearic acid or lyluic acid and Hg represent a benlyyl group. ) A method for producing an aconine alkaloid, which comprises obtaining an aconine alkaloid represented by:

■ General formula (in the formula, R1 is a lower alkyl group, R2 is a hydrogen atom or a hydroxyl group, R3 is a linole group, each acid residue of valmitic acid, oleic acid, stearic acid or linolenic acid, and Bz is a benryyl group) An anti-inflammatory analgesic containing an aconine alkaloid represented by

Claims (1)

[Claims] ■ General formula (wherein R□ is a lower alkyl group, R2Fi hydrogen atom or hydroxyl group, R3 is linoleic acid, pallic acid, oleic acid,
Each acid residue of stearic acid or lyluic acid and lJx represent a besilyl group. ) is an aconidial alkaloid. ■ The alcoholic extract of Huafuji is separated by the column column method and the general formula (in the formula, R□ is a lower alkyl group, R2 is a hydrogen atom or a hydroxyl group, R3 is linoleic acid, pallic acid, oleic acid) A method for producing aconidial alkaloid, which comprises obtaining an aconidial alkaloid represented by the following formula: each acid residue of stearic acid or lylisic acid, and 13z represents a benlyyl group. ■ General formula (in the formula, R□ is a lower alkyl group, R2 is a hydrogen atom or a hydroxyl group, R3 is each acid residue of linoleic acid, valmitic acid, oleic acid, stearic acid, or liluic acid, and Bz is a besidiyl group) ) An anti-inflammatory analgesic containing aconidial alkaloid.