JPH0222069B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel 1-substituted-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole derivatives and antithrombotic agents containing the derivatives as main ingredients. The compounds according to the invention have the general formula (In the formula, R means a linear or branched alkyl group having 1 to 5 carbon atoms, a phenyl group, or a substituted phenyl group substituted with a chlorine atom, nitro group, or methoxy group at the para position) The compound according to the present invention represented by the above general formula is a new substance that has not been described in any literature, and is It can be obtained by nitrosating a 1-substituted-2-[(N-morpholinoamino)methyl]-tetrazole represented by the formula (wherein R has the above-mentioned meaning). The compounds according to the present invention have antithrombotic effects and are therefore useful for the prevention and treatment of various diseases caused by the occurrence of thrombosis. When formulating the compound according to the present invention as an antithrombotic agent, there are no particular restrictions on the dosage form, and therefore, oral administration forms such as tablets, powders, fine granules, granules, capsules, and oral solutions can be used. Alternatively, it can also be made into an injection. In this case, formulation can be carried out by a conventional method. The dosage varies depending on the type of compound as the active ingredient, the severity of the disease, the dosage form, the age of the patient, etc., but when targeting adults,
Generally, the amount of the compound is 30-300 mg/day. Next, the present invention will be explained in more detail with reference to production examples, pharmacological test examples, and formulation examples. Production Example 1 a Synthesis of 1-phenyl-2-[(N-morpholinoamino)methyl]-tetrazole 7.8 g (0.040 mol) of 1-phenyl-5-chloromethyl-tetrazole was added to 30.9 g of N-aminomorpholine ( 0.303 mol), stirred for 3 hours at room temperature, added 30 ml of concentrated aqueous ammonia, and extracted with methylene chloride. The extract is dried over sodium sulfate, concentrated, and developed with ether using a silica gel column to obtain 3.0 g of the target substance in the form of an oil (yield 28.8%). IR spectrum (KBr) cm ^-1 : 3230 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.30 (methylene group) MS spectrum (CI/DI) : 261 (M+1) b 1-phenyl-2-[(N-morpholino-N
Synthesis of -nitrosamino)methyl]-tetrazole 1-phenyl-2- obtained by the method described in section a
Dissolve 1.93 g (7.4 mmol) of [(N-morpholinoamino)methyl]-tetrazole in 30 ml of water,
Add 0.74 ml of % hydrochloric acid dropwise. Cool this mixture on ice,
Sodium nitrite 0.53g (7.4 mmol) in water 13
ml and added dropwise to the above mixture and after stirring for 1 hour, the reaction mixture was extracted with methylene chloride, dried and concentrated. If recrystallized from methylene chloride/ether, the desired substance with a melting point of 101-3℃ can be obtained.
1.4 g is obtained (yield 65.4%). IR spectrum (KBr) cm ^-1
:1458 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:5.12 (methylene group) MS spectrum (CI/DI)
:261(M-28), 259(M-30) Production Example 2 a Synthesis of 1-p-nitrophenyl-2-[(N-morpholinoamino)methyl-tetrazole 1-p-nitrophenyl-5-chloromethyl-
16.0 g (0.067 mol) of tetrazole was cooled with N under ice cooling.
- Added to 34.0 g (0.334 mol) of aminomorpholine, then stirred at room temperature for 3 hours, and added to 100 ml of water.
Add 30 ml of concentrated ammonia water, and extract with chloroform. The extract is dried over sodium sulfate, concentrated, and developed with ether/ethyl acetate using a silica gel column to obtain 3.1 g of the target substance with a melting point of 135-7°C (yield 15.2%). IR spectrum (KBr) cm ^-1 : 3220 (amino group) NMR spectrum (chloroform-d ₁ ) δ
:4.33 (methylene group) MS spectrum (CI/DI): 306 (M+1) b Synthesis of 1-p-nitrophenyl-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole By the method described in section a The obtained 1-p-nitrophenyl-2-[(N-morpholinoamino)methyl]-
500 mg (1.7 mmol) of tetrazole is dissolved in 8 ml of water, and 0.17 ml of 36% hydrochloric acid is added dropwise thereto. This mixture was cooled on ice, 120 mg (1.7 mmol) of sodium nitrite was dissolved in 4 ml of water, and added to the above mixture, stirred for 1 hour, extracted with methylene chloride, and dried the extract with sodium sulfate. Concentrate. If recrystallized from methylene chloride/ether, melting point is 185.
400 mg of target substance at ~9°C is obtained (yield 73.0
%). IR spectrum (KBr) cm ^-1
:1440 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:5.03 (methylene group) MS spectrum (CI/DI)
:306 (M-28), 304 (M-30) Production example 3 a Synthesis of 1-p-methoxyphenyl-2-[(N-morpholinoamino)methyl]-tetrazole 1-p-methoxyphenyl-5 16.0 g (0.071 mol) of -chloromethyl-tetrazole is added to 36.0 g (0.356 mol) of N-aminomorpholine under ice-cooling, and the mixture is then brought to room temperature and stirred overnight. Add 50 ml of concentrated ammonia water and 100 ml of water to the reaction mixture,
Extract with chloroform. The extract is dried over sodium sulfate, concentrated, and developed with ether/ethyl acetate using a silica gel column to obtain 4.5 g of the target substance as an oil (yield 21.7%). IR spectrum (KBr) cm ^-1 : 3230 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.23 (methylene group) MS spectrum (CI/DI) : 291 (M+1) b Synthesis of 1-p-methoxyphenyl-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole As described in section a 3.0 g (9.8 mmol) of 1-p-methoxyphenyl-2-[(N-morpholinoamino)methyl]-tetrazole obtained by the method was added to 40 g of water.
ml, and 0.98 ml of 36% hydrochloric acid is added dropwise to this.
Keep this mixture below 5°C and add sodium nitrite.
A solution of 0.68 g (9.8 mmol) dissolved in 30 ml of water was added dropwise to the above mixture, and after stirring for 2 hours, it was extracted with methylene chloride. The extract was dried over sodium sulfate, concentrated, and diluted with methylene chloride/ether. When recrystallized, 2.2 g of the target substance with a melting point of 115-6° C. is obtained (yield 66.7%). IR spectrum (KBr) cm ^-1
:1440 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:4.95 (methylene group) MS spectrum (CI/DI)
:291(M-28), 289(M-30) Production example 4 a Synthesis of 1-p-chlorophenyl-2-[(N-morpholinoamino)methyl]-tetrazole 1-p-chlorophenyl-5-chloromethyl-
17.0g (0.0742mol) of tetrazole under ice cooling,
Add 37.9 g (0.371 mol) of N-aminomorpholine, then allow to come to room temperature and stir overnight. 50 ml of concentrated ammonia water was added to the reaction mixture, extracted with chloroform, the extract was dried over sodium sulfate, concentrated, developed with ether/ethyl acetate using a silica gel column, and further extracted with chloroform/ethyl acetate.
Recrystallization from ether yields 5.9 g of the target substance with a melting point of 100-1°C (yield 27.0%). IR spectrum (KBr) cm ^-1 : 3210 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.25 (methylene group) MS spectrum (CI/DI) : 295 (M+1) b Synthesis of 1-p-chlorophenyl-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole By the method described in section a The obtained 1-p-chlorophenyl-2-[(N-morpholinoamino)methyl]-
3.0 g (93 mmol) of tetrazole is dissolved in 30 ml of water, and 0.93 ml of 36% hydrochloric acid is added dropwise thereto. While cooling the resulting mixture on ice, add 0.67 g of sodium nitrite.
(93 mmol) dissolved in 10 ml of water was added dropwise to the above mixture, stirred for 1 hour, and then extracted with methylene chloride. The extract was dried over sodium sulfate, concentrated, and recrystallized from chloroform/ether. As a result, 2.2 g of the target substance with a melting point of 123°C (decomposition) is obtained (yield 73.3%). IR spectrum (KBr) cm ^-1
:1440 (Nitroso group) NMR spectrum (DMSO-d ₆ ) δ
:5.20 (methylene group) MS spectrum (CI/DI)
:295(M-28), 293(M-30) Production example 5 a 1-methyl-2-[(N-morpholinoamino)
Synthesis of methyl]-tetrazole 1-methyl-5-chloromethyl-tetrazole
13.5 g (0.102 mol) was added to 78.5 g (0.770 mol) of N-aminomorpholine under ice cooling, and then the mixture was allowed to reach room temperature and stirred for 13 hours. 60 ml of concentrated ammonia water was added to the reaction mixture, extracted with chloroform, the extract was dried over sodium sulfate, concentrated, and purified with ethyl acetate/n- using a silica gel column.
When developed with hexane/ethanol, 3.0 g of oily target substance is obtained (yield 15.0%). IR spectrum (KBr) cm ^-1 : 3230 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.26 (methylene group) MS spectrum (CI/DI) : 199 (M+1) b 1-Methyl-2-[(N-morpholino-N-
Synthesis of nitrosamino)methyl]-tetrazole 1-methyl-2- obtained by the method described in section a.
1.68 g (8.5 mmol) of [(N-morpholinoamino)methyl]-tetrazole is dissolved in 10 ml of water, and 0.87 ml of 36% hydrochloric acid is added dropwise thereto. Cool this mixture on ice and use 0.60 g (8.7 mmol) of sodium nitrite.
A solution of dissolved in 5 ml of water was added dropwise to the above mixture,
Stir for 1 hour, extract with methylene chloride, dry the extract with sodium sulfate, concentrate, and recrystallize with methylene chloride/n-hexane.
1.38 g of the target substance at 9°C is obtained (yield 72.0%). IR spectrum (KBr) cm ^-1
:1445 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:5.10 (methylene group) MS spectrum (CI/DI)
: 199 (M-28), 197 (M-30) Production example 6 a 1-ethyl-2-[(N-morpholinoamino)
Synthesis of methyl]-tetrazole 1-ethyl-5-chloromethyl-tetrazole
12.0 g (0.082 mol) is added to 41.8 g (0.410 mol) of N-aminomorpholine under ice cooling, brought to room temperature, and then stirred overnight. 50 ml of concentrated aqueous ammonia and 50 ml of saturated brine were added to the resulting reaction mixture, extracted with methylene chloride, dried over sodium sulfate, concentrated, and then developed with ether/ethyl acetate using a silica gel column. , melting point
2.3g of target substance at 91-3℃ is obtained (yield 13.2
%). IR spectrum (KBr) cm ^-1 : 3230 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.30 (methylene group) MS spectrum (CI/DI) : 213 (M+1) b 1-ethyl-2-[(N-morpholino-N-
Synthesis of nitrosamino)methyl]-tetrazole 1-ethyl-2- obtained by the method described in section a.
2.0 g (94 mmol) of [(N-morpholinoamino)methyl]-tetrazole is dissolved in 10 ml of water, and 0.94 ml of 36% hydrochloric acid is added dropwise thereto. This mixture was cooled on ice, and a solution of 0.67 g (94 mmol) of sodium nitrite dissolved in 4 ml of water was added dropwise to the above mixture, stirred for 1 hour, extracted with methylene chloride, and the extract was dried over sodium sulfate. If concentrated, then recrystallized from methyl ethyl ketone/ether, the melting point
1.3g of target substance at 50-1°C is obtained (yield 60.0
%). IR spectrum (KBr) cm ^-1
:1450 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:5.04 (methylene group) MS spectrum (CI/DI)
: 213 (M-28), 211 (M-30) Production example 7 a Synthesis of 1-isopentyl-2-[(N-morpholinoamino)methyl]-tetrazole 1-isopentyl-5-chloromethyl-tetrazole 14.0 g (0.074 mol) was added to 38.0 g (0.373 mol) of N-aminomorpholine under ice cooling,
It is then brought to room temperature and stirred overnight. 50 ml of concentrated aqueous ammonia and 50 ml of saturated brine were added to the resulting reaction mixture, extracted with methylene chloride, the extract was dried over sodium sulfate, concentrated, and then diluted with ether/ethyl acetate using a silica gel column. 2.4g of oily target substance is obtained (yield:
12.7%). IR spectrum (KBr) cm ^-1 : 3230 (amino group) NMR spectrum (chloroform-d ₁ ) δ
: 4.27 (methylene group) MS spectrum (CI/DI) : 255 (M+1) b Synthesis of 1-isopentyl-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole Obtained by the method described in section a. 2.0 g (79 mmol) of 1-isopentyl-2-[(N-morpholinoamino)methyl]-tetrazole is dissolved in 20 ml of water, and 0.79 ml of 36% hydrochloric acid is added dropwise thereto. While cooling this mixture on ice, a solution of 0.56 g (79 mmol) of sodium nitrite dissolved in 4 ml of water was added dropwise to the above mixture, and the mixture was stirred for 1 hour. The resulting reaction mixture is extracted with methylene chloride, the extract is dried over sodium sulfate, concentrated, and crystallized from methyl ethyl kentone/ether to obtain 1.4 g of the target substance with a melting point of 50-1°C. (yield 61.6%). IR spectrum (KBr) cm ^-1
:1450 (nitroso group) NMR spectrum (chloroform-d ₁ ) δ
:5.04 (methylene group) MS spectrum (CI/DI)
:213 (M-28), 211 (M-30) The elemental analysis results of the nitroso compounds obtained in each of the above production examples were as follows.

【table】

[Table] Pharmacology test examples [] Action on vascular smooth muscle a Experimental method Mesenteric arterial blood vessels were removed from rabbits, and the connective tissue and adventitia were immediately removed using the method of Lewis et al.
J.Hand Koessler, KK “Arch.Intern.Med.” No.
39, pp. 182-7, 1927).
A test strip is prepared by cutting it spirally at a 45° angle.
A test strip of an appropriate length was suspended in a Krebs-Henseleite nutrient solution kept at 37°C while aerating 95% O ₂ -5% CO ₂ gas. acement) and record the isometric tension changes with a recorder. Allow test strip specimens to equilibrate for at least 1 hour prior to beginning the experiment. During this time, a constant tension (1 g) is applied in advance. First, KCl and CaCl ₂ , serotonin, histamine,
Norepinephrine or the like is added to contract the strip to approximately 40-60% of its maximum contraction, and after the tension becomes constant, administration of the drug is started. Drugs are administered cumulatively at approximately three times the concentration (1×10 ^-8 M, 3×
10 ^-8 M, 1×10 ^-7 M, 3×10 ^-7 M…). lastly
Administer 10 ^-4 M papaverine to induce relaxation.
The degree of relaxation at each concentration is expressed as %, with 100%. The horizontal axis shows the drug concentration, and the vertical axis shows the degree of relaxation (%).
and calculate the 50% relaxation concentration (ED ₅₀ ). b Experimental Results The results were as shown in Table 1 below. Table 1 Nitroso compounds (manufacturing examples) ED ₅₀ (μM) 1 0.8 2 0.15 3 0.13 4 0.09 5 0.11 6 0.10 7 0.14 [] Effect on human platelet aggregation a Experimental method Citrated blood (0.38% sodium citrate vs. human blood) Using platelet-rich plasma (PRP) prepared from 1:9), the method of Born et al. (Born, GVR "Nature"
194, pp. 927-9, 1962), platelet aggregation was measured. Sample solutions of various concentrations were prepared by diluting the compound according to the present invention with physiological saline, and 0.03 ml of the sample solution was diluted with physiological saline.
Add to 0.27 ml of PRP, incubate for 3 minutes at 37°C, then add appropriate amounts of various aggregation-inducing substances, and replace the resulting aggregation with the light transmittance process, that is, the light transmittance when PRP is 0 to 100%. Record. b Experimental Results The results were as shown in Table 2 below.

[Table] [] Acute toxicity a Experimental method A constant temperature and humidity breeding room with a room temperature of 23 ± 1°C and a humidity of 55 ± 5% was used.
g) were divided into 6 groups as test animals, and among the compounds according to the present invention, the compound obtained in Production Example 1 was used as a representative test substance, and this was suspended in a 5% gum arabic solution. The drug is orally administered to animals, followed by observation for one week, and the acute toxicity value is determined from the mortality rate. b Experimental Results The results were as shown in Table 3 below.

[Table] Formulation Example Tablets were manufactured using the following ingredients in a conventional manner (1000 tablets). Nitroso compound (Production Example 1) 30.0 (g) Crystalline cellulose 15.6 Carboxymethyl cellulose (Ca) 8.4 Corn starch 12.0 Lactose 53.4 Magnesium stearate 0.6 120.0 (g)

Claims (1)

[Claims] 1. General formula (In the formula, R means a linear or branched alkyl group having 1 to 5 carbon atoms or a phenyl group, or a substituted phenyl group substituted at the para position with a chlorine atom, nitro group or methoxy group) A novel 1-substituted-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole derivative shown in 2 General formula (In the formula, R means a linear or branched alkyl group having 1 to 5 carbon atoms or a phenyl group, or a substituted phenyl group substituted at the para position with a chlorine atom, nitro group or methoxy group) An antithrombotic agent containing a novel 1-substituted-2-[(N-morpholino-N-nitrosamino)methyl]-tetrazole derivative as a main component.