JPS63132831A - Antiserotonin agent - Google Patents

Abstract

PURPOSE:To obtain an antiserotonin agent, containing a cyclohexane derivative as an active ingredient, having serotonin antagonistic action as well as low toxicity and useful for treating depression, schizophrenia, hemicrania, hypertension, peripheral cardiovascular disorder, etc. CONSTITUTION:An antiserotonin agent obtained by blending a cyclohexane derivative expressed by the formula (R is alkyl; R<1> and R<2> are H or organic residue) as an active ingredient with a carrier, excipient, diluent, etc., and formulating the resultant blend by a conventional method. The dose thereof is 0.1-1.5mg as the compound expressed by the formula per day for an adult administered once - in several divided portions. The compound expressed by the formula is obtained by adding ethyl ether to a hot water extract of cinnamon which is a herb medicine, distributing and fractionating the extract by a well-known method.

Description

[Detailed description of the invention] [Industrial application field] The present invention relates to novel antiserotonin agents.

[Prior art/problems to be solved by the invention] Serotonin (i.e., 5-hydroxytryptamine) is one of the autocoids (active substances in the living body) produced in the living body, and is used in platelets, serum, lungs, and lI. It is widely distributed in the walls and central nervous system (especially the hypothalamus, linea, medulla oblongata, pons, amygdala, etc.).

The pharmacological actions of serotonin include smooth muscle contraction (i.e., contraction of blood vessels, bronchi, ureters, and nictitating membranes, smooth muscle contraction of the intestinal tract and uterus, etc.), promotion of secretion, effects on the central nervous system, and effects on afferent nerves. It is known to have effects such as increased sensitivity to sensory stimuli, elicitation of reflexes from receptors, excitation of sensory nerve endings, histamine release, etc., and effects on the circulatory system (vasoconstriction or relaxation, myocardial stimulation, etc.).

Anti-serotonin drugs have an antagonistic effect on serotonin, so they are clinically used to treat depression, schizophrenia, migraines,
It is thought to be useful in the treatment of hypertension, peripheral circulation disorders, carcinoid syndrome, allergic diseases, etc.

On the other hand, cinnamon (Cinnamomum, Lauraceae, Cinna brume) has been traditionally used as a Chinese herbal medicine.
momua +, cassia Blume) bark]
It has been found that the water-soluble extract fraction of 1999 has anti-ulcer activity (Japanese Patent Application Laid-Open No. 59-65018), and the present inventors have discovered that one of the active substances is 2-(2-acetylvinylidene)- 1
,3,3-)Samethylcyclohexane-1,5-diol-5-〇-β-D-apiofuranosyl(l→6)-
It has been found that it is β-D-glucopyranoside [Compound (T-1)] (Japanese Patent Application No. 112837-1983).

[Problem that the invention seeks to solve]

The present inventors aim to provide a medicament that is attributable to pharmacological activities other than anti-ulcer activity of this compound (+-1) and compounds related thereto.

[Means for solving problems]

From this point of view, the present inventors have conducted extensive research on the pharmacological activity of the above-mentioned compound, and as a result found that the compound has a frame serotonin antagonistic effect, and after further research, they have completed the present invention.

The present invention relates to a cyclohexane derivative represented by the general formula (wherein R is alkyl, and R1 and R2 each represent a hydrogen atom or an organic residue) [hereinafter referred to as compound (1)]
)] is an active ingredient.

The active moiety in the antiserotonin agent of the present invention is a moiety represented by the formula: Therefore, the organic residue used herein is not particularly limited as long as it is pharmacologically acceptable. Examples of such organic residues include the following. That is, examples of R1 include alkyl, acyl, alkoxycarbonyl, alkoxycarbonylalkyl, carboxyalkyl, carboxyalkylcarbonyl, and cyclic acetal.

Regarding R8, oligosaccharide residues of sugar units 1 to 3,
Examples include alkyl, acyl, alkoxycarbonyl, alkoxycarbonylalkyl, carboxyalkyl, carboxyalkylcarbonyl, and cyclic acetal.

In this specification, alkyl preferably has 1 to 6 carbon atoms, particularly 1 to 4 carbon atoms, and may be either linear or branched, specifically methyl, ethyl, n-propyl, 1so- propyl, n-butyl, t-butyl, n
-Pentyl etc. are exemplified. Furthermore, the alkoxy group preferably has 1 to 6 carbon atoms, particularly 1 to 3 carbon atoms, and may be either linear or branched, and specifically includes methoxy, ethoxy, n-propoxy, 1so-propoxy,
Examples include n-butoxy and t-butoxy. moreover,
The acyl may be either aliphatic or aromatic,
Examples of the aliphatic type include those having 1 to 6 carbon atoms, particularly 2 to 5 carbon atoms, such as acetyl, propionyl, butyryl, and valeryl, and examples of the aromatic type include benzoyl.

Further, the alkoxy moiety in alkoxycarbonyl is as described above, and examples of the group include methoxycarbonyl, ethoxycarbonyl, and the like. The alkoxy moiety and alkyl moiety in alkoxycarbonylalkyl are as described above, and the group includes:
For example, methoxycarbonylmethyl is exemplified. The alkyl moiety in carboxyalkyl is as described above, and examples of the group include carboxymethyl. The alkyl moiety in carboxyalkylcarbonyl is as described above, and an example of the group is carboxyethylcarbonyl. Examples of the cyclic acetal include tetrahydropyranyl.

Furthermore, the oligosaccharide residues of sugar units 1 to 3 refer to mono-, di- or tri-glycoside residues. The constituent sugars are not particularly limited, and specifically, monoglycoside residues include glucosyl groups, arabinosyl groups, galactosyl groups, mannosyl groups, fructosyl groups, xylosyl groups, ribosyl groups, apiosyl groups, glucosamine groups, and diglycoside residues. Examples of triglycoside residues include apiosyl glucosyl group, scrosyl group, maltosyl group, lactosyl group, and gentiobiosyl group; examples of triglycoside residues include apiosyl gentiobiosyl group, gentianosyl group, and ruffinosyl group. The hydroxyl groups of these sugar moieties may be partially or entirely substituted with lower alkyl, acyl, alkoxycarbonyl, alkoxycarbonylalkyl, carboxyalkyl, carboxyalkylcarbonyl, or cyclic acetal.

This compound (1) can be obtained, for example, as follows.

First, an aqueous solution fraction derived from cinnamon is prepared by a known method (for example, JP-A-59-65018).

That is, ethyl ether is added to a hot water extract of cinnamon to perform partitioning and fractionation, and a water-soluble fraction is recovered.

The hot water extracted fraction of cinnamon was subjected to normal phase adsorption chromatography (silica gel, activated alumina, etc. as a packing material, chloroform + methanol, chloroform + methanol decahydrate, ethyl acetate + ethanol decahydrate, hexane + acequine, etc. as an eluent). , reversed-phase partition chromatography [filling materials such as C11 (octadecylsilane), Off (octylsilane), etc., elution solvents such as methanol tap water, acetonitrile decaqueous, tetrahydrofuran decaqueous, etc.], other chromatography (amber as a packing material) A compound represented by the formula [compound (1-1)] is obtained by fractionation using an appropriate combination of light XAD-2 (eluent such as methanol and tap water).

By hydrolyzing this compound (1-1) by a conventional method, a compound represented by the formula [compound (r-2)] is obtained.

The compound (1-1) or compound (1-2) can be derivatized by a conventional method to form another compound (+).

For example, oligo I of compound (■-2)! The derivative is produced by reacting compound (1-2) with an activated glycoside (eg, bromoglycoside).

At that time, it is preferable to carry out the reaction in the presence of a solvent inert to the reaction (e.g., chloroform, dichloromethane, tetrahydrofuran, etc.), the reaction temperature is about 0°C to 80°C, and the reaction time is about 0.5 to 5 hours. is preferred.

In addition, by treating the compound (r-1) by means known per se, the general formula (wherein R and R1 have the same meanings as above, R1, R4, R
', R'', R7 and R8 are each an organic residue, preferably alkyl, acyl, alkoxycarbonyl, alkoxycarbonylalkyl, carboxyalkyl, carboxyalkylcarbonyl, cyclic acetal, etc. [Compound H-3] ) is manufactured.

For example, in compound (1-3), R is methyl, R
A compound in which 1, R, R4, R1, R1, R7 and R1 are each alkyl is produced by alkylating the compound ([-1) by a conventional method.

Preferred alkylating agents in the alkylation include alkyl halides (eg, methyl iodide, etc.), alkyl sulfates (eg, dimethyl sulfate), and the like. The alkylation is preferably carried out in the presence of a base (e.g. sodium hydride,
One reaction time carried out in the presence of silver oxide, potassium carbonate, sodium carbonate, etc.) is about 0.5 to 5 hours, and the reaction temperature is about θ°C to 80°C.

Further, R is methyl, R1, R3, R4, 8%, Rh.

A compound in which R' and R8 are each acyl is produced by acylating compound (1-1). The acylating agent in the acylation preferably includes carboxylic acids and reactive derivatives thereof. Examples of the reactive derivatives include those known per se, such as acid halides (eg, acid chloride, acid bromide), acid anhydrides, and active esters. When using a carboxylic acid as the acylating agent, it is preferable to carry out the reaction in the presence of a condensing agent such as dicyclohexylcarbodiimide, and when using other acylating agents, it is preferable to carry out the reaction in the presence of a tertiary condensing agent such as pyridine or triethylamine. It is preferable to carry out the reaction in the presence of an amine, and in any case, it can also be carried out in the presence of a solvent inert to the reaction (chloroform, dichloromethane, tetrahydrofuran), etc. The reaction time is 0.5 to 5 hours. The reaction temperature is 0°C to 25°C.
That's about it.

Compound (1) obtained in this way is subjected to dissolution, recrystallization,
It can be isolated and purified by any conventionally known method such as chromatography.

Next, the methods and results of experiments conducted to confirm the pharmacological effects, acute toxicity tests, administration methods, etc. of the compound of the present invention will be shown.

(Pharmacological effect) m Central serotonin antagonism 5-hydroxytryptophan (5
The effect was determined using the number of head convulsions (light exposure for 5 minutes) induced by -HTP) 200 μ/kg as an index. Test drug (compound (+-1)) and physiological saline (
5Pl) were administered into the fourth ventricle immediately after administration of 5-HTP. The dose of compound (I-1) was 0.3 pg/
rats, 4 rats per group.

(Margin below) ” p < o, os °P < 0.01 ””
r'<0.001Student t-test
(against control value)
(2) Central serotonin antagonism DL-5HTP200/ktr was subcutaneously administered to the back of male ddY mice (body weight 25-30 g) to induce head spasticity, and the number of head spasticity produced in 10 minutes was determined. The effectiveness was determined using this as an indicator. Test drug [compound (1-2)) and physiological saline (10a7/kg each)
(Administered orally 1 hour before TP administration. Compound (1-2
) dose was 6.5 pg/kg, 8 animals per group.

(Results) a. Regarding the effect on JR@convulsion-induced response latency, as described above, compound (1-2) delayed the convulsion-induced response by about 90 seconds.

b, Number of convulsions in the g region As mentioned above, the number of convulsions vI induced was approximately 33 for the compound (+-2).
% suppressed.

(3) Peripheral serotonin antagonism Wistar male rats (weighing 200-300 g) were killed by talking, the stomach was removed, and the gastric contents were washed. 3+nX1.5
Cut into cm rectangles. This specimen was placed in a lO− water bath (
The muscle contractions were captured using a force displacement transducer and recorded on a recording needle. The nutrient solution in the water bath was Krebs' solution, and 95% O and +5% COt were aerated therein. A solution of serotonin creatine sulfate in distilled water was used as a smooth muscle contraction stimulant, and the concentration in a water bath (final concentration) was 10-9 to 101 M.
Samples were used up to.

The test drug [compound (I-1)] was applied at a dose of 0.1 m into the water bath immediately before serotonin administration. Each contraction height was expressed as a ratio to the maximum number contraction in these dose ranges of serotonin as 100%.

The amount of compound (1-1) added (final Ilk concentration) was 30 nM.

(Results) As shown in FIG. 1, the dose-response curve was shifted to the right, suggesting that serotonin antagonism occurred.

(4) Peripheral serotonin antagonism method is based on (3) Compound (+-1). Compound (1
-2) The added N (final concentration) was 3.30.300 nM.

(Results) As shown in FIG. 2, the dose-response curve was shifted to the right, suggesting that serotonin antagonism occurred.

(Toxicity) In an acute toxicity experiment using the compound (1) of the present invention using ddYi mice, 25
No lethal toxicity was observed even at doses up to 0 kg/kg.

(Dosage and administration method) The antiserotonin agent containing the compound (H) of the present invention as an active ingredient can be administered to mammals (humans, horses, dogs, mice, rats, etc.)
It is useful for depression, schizophrenia, migraines, vascular headaches, peripheral circulation disorders, hypertension, carcinoid syndrome, dumping syndrome, allergic diseases, etc.

The antiserotonin agent of the present invention can be administered orally or parenterally, but in the case of oral administration, compound (1) is mixed with appropriate pharmaceutically acceptable additives (carriers, excipients, diluents, etc.), and a powder is prepared. It is used as tablets, capsules, troches, solutions, syrups, and granules. In the case of parenteral administration, it is used as an aqueous solution or non-aqueous suspension, an injection such as intravenous injection, intramuscular injection, or subcutaneous injection, or a suppository.

The dosage may vary depending on the patient's symptoms, body weight, age, etc., but it is usually 0.1 to 0.1 to 0.1 to 0.1 as compound (1) per day for adults.
The dose is preferably 1.5 μm, which is administered once or divided into several doses.

[Action/Effect]

The anti-serotonin agent consisting of compound (1) has an extremely excellent serotonin antagonistic effect, has extremely low toxicity, and exhibits remarkable pharmacological effects.

〔Example〕

Example 1: Tablet + 11 compounds (1) 0.5■
(2) Fine particles for direct impact m 209 (manufactured by Fuji Chemical Co., Ltd.)
46.6 ■Magnesium aluminate metasilicate 20% corn starch 30% milk @
50% (3) Crystalline cellulose 24. ON+41 CM C Calcium 4. OLIf (5) Magnesium stearate 0.4 ■ (11) Pass all of (3) and (4) through a 100-mesh sieve in advance, and dry this full, (3), (4), and (2), respectively. After lowering the moisture content to a certain level, mix using a mixer at the above weight ratio.Add (5) to the evenly distributed mixed powder, mix for a short time (30 seconds), and mix the mixed powder. Locks (Pestle: 6
．． 3-Kasumi φ, 6.0 Dragon R) to make tablets each weighing 75.5 cm.

The tablets may be coated with a commonly used gastric soluble film coating agent (eg, polyvinyl acecure diethylamino acetate) or an edible coloring agent, if necessary.

Example 2: Capsule (1) Compound N) 2.5 g (
2) Milk 11 935 g
(3) Magnesium stearate 15
(g) The above components were weighed and mixed in a backward direction, and the mixed powder was filled into hard gelatin capsules in an amount of 190 square centimetres.

Example 3: Injection fi + compound (1) 0.5■
(2) Grapes ¥9 100
■(3) Physiological saline 10
After filtering the above mixture with a membrane filter, sterilization filtration was performed again, and the filtrate was aseptically dispensed into vials, which were filled with nitrogen gas and sealed to give an intravenous injection.

Reference Example 1: Preparation of Compound (1-1) Hot water at 60 to 100°C is applied to 50 kg of cinnamon for 1 to 2 hours, and the resulting extract is adsorbed onto 100 to 140 m+ of Amberlite XAD-2 resin.

After washing the resin with water, using a 40-80% methanol aqueous solution as an eluent, the resulting eluate fraction was concentrated under reduced pressure and then freeze-dried in vacuo to obtain 148 g of crudely purified powder. From this crudely purified powder, preparative normal phase high performance liquid chromatography (
Compound (1-1) was obtained as a single peak component by sequentially repeating fractionation by both HPLC (C1) and preparative reverse-phase HPLC using 6 to 10% acetonitrile aqueous solution as a mobile phase solvent in the column. , that is, 2-(2-acetylvinylidene')-1,3,3-()tumethylcyclohexane-1,5-diol-5-〇-β-D-apiofuranosyl(1-6)-β- D-glucopyranoside 4
1.05■ was obtained.

The physical properties of compound (1-1) were as follows.

Properties: White amorphous powder [α)! ": -88,6" (MeOH,cO
, 5%) UV (MeOH): λwax 23I
ns (log = 4.07)IR(neat,
cm-'): 3350.2930,1940,1665,1610.
864,821' HN M R (MeOH-Da,
δ111111) :5.83 (18,s).

5.00 (IH, d, J = 2.4Hz).

4.42 (LH, d, J = 7.9Hz).

4.31 (IH, w+).

3.98 (2■+1lL 3.89 (IH, d, J = 2.4Hz).

3.77 (1B, d, J = 9.7Hz).

3.62 (LH, dd, J=5.5. 11.
4Hz).

3.58 (2H, s).

3.45-3.39 (IH, m).

3.36-3.25 (2) 1. Fog).

3.14 (1)1. dd, J=7.9.
8.4Hz).

2.35 (IH, d, J = 14.2Hz).

2.19 (3H, s).

2.13 (LH, d, J=14.2Hz).

1.52-1.43 (2H, s).

1.40 (3H, s).

1.39 (3H, s).

1.16 (3)1. s) '3CNMR(Meal(-Da, δpp+*
) :211.9(s), 201.2(s),
120.8(s), 111.4(d).

103.6(d), 101.7(d), 81.0(
s), 78.7(d).

78.7(d), 77.3(d), 75.7(
d), 75.6(t).

73.7(d), 72.9(s), 72.4(
d), 69.1(t).

66.6(t), 48.8(t), 47.4(
t), 37.5(s).

32.8(q), 31.3(q), 30.0(
q), 27.0(q) Reference Example 2: Compound (1-2)
Preparation of 20 ml of compound (1-1) was dissolved in a mixture of 2-l methanol and 2N hydrochloric acid, and reacted at 39°C for about 6 hours. After concentrating the reaction solution under reduced pressure, the reaction solution was purified and fractionated using preparative reverse-phase HPLC using a C1s column and a 6 to 10% acetonitrile aqueous solution as a mobile phase solvent to obtain a yellow oily aglycone compound (+-2), part 4-(2,4-dihydroxy-2,6゜6-drimethylcyclohexylidene)-
7.5 parts of 3-buten-2-one were obtained.

I R (neat, cm-'): 3620, 2920.1945.167'0' H-N
M R(MeOH-D4. δI'P11+ 2
00MHz): 5.80 (IH, s).

4.28 (IH, +*).

2.40-2.10 (2H, m).

2.15 (3H, s).

1.50-1.45 (2H, m).

1.40 (38,s).

1.36 (3H, s).

1.16 (3H, s) Reference Example 3 Preparation of near-glycone derivative (glucose glycoside) Compound (1-2) 4.5■, 2. 3. 4. 6-tetra-0-acetyl-α-D・-bromohexose 12
．． A mixture of 5 ml of chloroform and 0.5 st 7 of a 1.25N aqueous sodium hydroxide solution containing 5 ml of benzyltriethylammonium bromide and 8.5 ml of benzyltriethylammonium bromide was heated under reflux on a 60° C. oil bath for about 3 hours. After the reaction, the chloroform phase was concentrated and purified and fractionated on a silica gel column using a preparative normal phase system PLO using a mixed solvent of chloroform-methanol-water (40 to 70:10:1) as the mobile phase. 9.6 ml of tetraacetyl glucoside was obtained. Furthermore, dry methanol-benzene (2:1) 2.5aZ of this acetyl form
After adding 5 μ of water-cooled anhydrous potassium carbonate to the solution, it was stirred at room temperature for about 1.5 hours, and the reaction solution was concentrated under reduced pressure.
Purification and fractionation was performed by preparative reverse-phase HPLC using a 6-10% acetonitrile aqueous solution as a mobile phase in a column to obtain 5.7 mg of a monoglycoside compound as a white amorphous powder.

The physical properties of this monoglycoside are as follows.

I R (neat, cm-'): 3400, 2940.1945.1670.1600'
HNMR(MeOH-Da, δppm>:
5.81 (III, s).

4.40 (18, d, J=8Hz).

4.29 (1) 1. ■).

4.00 (1) 1. d, J=11; 2Hz).

3.63 (IH, dd, J=5.2.11.2Hz
).

3.45-3.25 (38,s).

3.15 (1B, dd, J-8,8,4H
z).

2.35 (1B, d, J = 14.1Hz).

2.18 (311, s).

2.12 (IL d, J=14.IHz).

1.55-1.45 (2H, n+).

1.40 (311, s).

1.37 (311, s).

1.16 (3H, s) Reference Example 4: Preparation of hebutaacetate of compound ([-1) Compound (1-1) 12■ was reacted with acetic anhydride 2@1 and pyridine 4I117 for 30 minutes at room temperature After concentrating the liquid under reduced pressure, it was purified and fractionated on a silica gel column using preparative normal-phase HPLC using a mixed solvent of chloroform-methanol-water (40-70:10:1) as the mobile phase, and a pale yellow oil was obtained. 12.2 ml of hebutaacetate was obtained.

IR (neat, Cm-'): 1940, 1750.1670.1600 (Of (no absorption observed)'H-NMR (MeOH-Ro4. δppm):
2.00 - 2.07 (3Hx 7.s, Ac) Reference Example 5: Preparation of hebutamethylate of compound (1-1) Under a nitrogen atmosphere, 25 μm of sodium hydride was dissolved in 70 μg of dimethyl sulfoxide. Stir at ℃ for about 1 hour, then cool to room temperature. In this solution, compound (1-1)
A solution of 8W dissolved in dimethyl sulfoxide 0.5@I was added, and after stirring for 1 hour, methyl iodide 0.1-
was added and stirred for an additional 4 hours. Approximately 5 a7 grams of ice water was added to this reaction solution, and then extracted twice with three portions of chloroform. The extract was concentrated and purified and fractionated using the preparative normal phase HPLC described above to obtain 7 parts of yellow oily hebutamethylate.

r R (neat, cm-'): 1945, 1660.1610.1580 (O)l absorption not observed)' HN MR (Meaty-Da, δPpII
): 3.34~3.95 (3Hx 7.s, O
Me)

[Brief explanation of the drawing]

Figure 1 shows a concentration response curve with serotonin concentration on the horizontal axis and muscle fiber contraction rate on the vertical axis. The solid line represents the physiological saline addition group (control), and the dotted line represents the compound (1-1 ) represents the addition group. FIG. 2 also shows a concentration-response curve in which the horizontal axis represents the serotonin concentration and the vertical axis represents the contraction rate of muscle fibers. -・- indicates the saline addition group (control), others indicate compound (1-2)
Represents the addition group. Procedural Amendments (issued by E.) April 1, 1986

Claims (1)

[Claims] A cyclohexane derivative represented by the general formula ▲ Numerical formulas, chemical formulas, tables, etc. An anti-serotonin agent containing as an active ingredient.