JPS6258336B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a phosphodiesterase inhibitor containing a novel tetrazolylpropoxycarbostyryl derivative as an active ingredient. The tetrazolylpropoxycarbostyryl derivative contained as an active ingredient in the phosphodiesterase inhibitor of the present invention has the general formula [In the formula, R represents a lower alkyl group or a cycloalkyl group]. Conventionally, the compound described in JP-A-54-30183 has been known as a compound having a structure similar to the compound of general formula []. The compounds described in the above-mentioned publication have antibacterial effects, anti-inflammatory effects, platelet aggregation inhibiting effects, and effects of lowering the blood concentration of lipid-containing substances in the blood, especially cholesterol, phospholipids, triglycerides, etc., and are antibacterial agents and anti-inflammatory agents. It is known as a blood clot preventive drug and a treatment and prevention drug for arteriosclerosis. However, as a result of various studies, the present inventors discovered that the compound of the general formula [] has a phosphodiesterase inhibitory effect that was completely unexpected from the use of the compound described in the above-mentioned publication, and completed the present invention. That is, the present invention relates to a phosphodiesterase inhibitor characterized by containing a compound of the general formula [] as an active ingredient. The phosphodiesterase inhibitor of the present invention is cyclic adenosine monophosphate (hereinafter referred to as
It has specific inhibitory activity against phosphodiesterase, which decomposes G-AMP (abbreviated as G-AMP). Substances that exhibit inhibitory activity against phosphodiesterase (hereinafter abbreviated as PDE), which is a C-AMP degrading enzyme, are described in, for example, the Annual Review of Pharmacology and Toxicology.
Toxicology) Volume 17, pp. 441-477 (1977)
As described in , it is useful for the prevention or treatment of various diseases in which a decrease in C-AMP is observed due to metabolic abnormalities. That is, in general,
Intracellular C-AMP is mediated by the sympathetic nervous system transmitters catecholamines and various peptide hormones via adenylate cyclase.
It is known to be a biological component produced from adenosine triphosphate (ATP) and degraded by PDE. C-AMP not only transmits the effects of biogenic amines and peptide hormones into cells, but also contributes to cell division, fertilization, embryonic growth and differentiation, smooth muscle tone, cardiac contraction and metabolism, and central and autonomic nervous system control. C-AMP metabolic abnormality (reduction) affects C-AMP system function, immune response, reproductive function, release of substances contained in intracellular storage granules such as insulin, histamine, and serotonin, and lysosomal enzyme system. C-AMP is closely related to various diseases such as cancer development, bronchial asthma, diabetes, arteriosclerosis, circulatory failure, hypertension, psychosis, and psoriasis. Effective in treating or preventing disease. Among PDE inhibitors, those that have a specific effect on C-AMP are particularly effective, as described in, for example, Molecular Pharmacology, Vol. 13,
400-406 (1976). Therefore, the PDE inhibitor of the present invention is useful as a prophylactic and/or therapeutic agent for various diseases such as those listed below. Advances in Cyclic Nucleotide Research
Nucleotide Research) Volume 1, Page 175 (1972
) and Molecular Pharmacolngy, Vol. 6, p. 597 (1970), it is stated that when a PDE inhibitor acts on vascular smooth muscle, its C-AMP content increases.
It has been described that the smooth muscle is expanded and the blood flow condition can be improved. This shows that PDE inhibitors are useful as circulation-improving drugs, especially as cerebral circulation-improving drugs. In spontaneously hypertensive rats (SHR), C-AMP content is decreased in diseased arteries, and this is reported to be due to increased PDE activity and attenuation of adenylate cyclase activity in response to vasodilatory stimuli [ Science No. 179
Volume, No. 807 (1973)]. This is thought to worsen the relaxation response of vascular smooth muscle. Therefore, inhibiting PDE and increasing C-AMP can attenuate vascular resistance and treat hypertension. Furthermore, in cases such as renal hypertension, the use of diuretics reduces the total blood volume and exerts a hypotensive effect. Known diuretic benzothiazide derivatives include:
It has a PDE inhibitory effect [Annals of New York Academy of Science]
150, p. 256 (1968)], and C.
- Increase in AMP enhances diuresis (Journal of Clinical Investigation, No. 41)
Vol., p. 702 (1968)]. PDE inhibitors are thus useful as antihypertensive agents and diuretics. Proceedings of the National
Academy of Sciences of the United States of America (Proceedings
of the National Academy of Science of
As described in United States of America, Vol. 68, p. 425 (1971), theophylline, a known PDE inhibitor, has the effect of normalizing cancerous and tumorigenic cells. This shows that the PDE inhibitor of the present invention is effective as a cancer therapeutic agent. Journal of Clinical Investigation
Investigation) Volume 52, Page 48 (1973),
Relaxation of bronchial smooth muscle has been noted to increase in C-AMP content due to the β-action of catecholamines, which are sympathetic neurotransmitters, and the C-AMP content in bronchial smooth muscle of patients with bronchial asthma generally tends to decrease. It is written that it shows. PDE inhibitors inhibit the degradation of C-AMP, thereby making it possible to increase the content of C-AMP and thereby relax bronchial smooth muscle. In fact, theophylline, a known PDE inhibitor, is known as a therapeutic agent for bronchial asthma due to this mechanism of action.
Therefore, the PDE inhibitor of the present invention is also useful as a therapeutic agent for bronchial asthma. Journal of Immunology
As reported in Immunology, Vol. 108, p. 695 (1972), allergic asthma patients
Generally, bronchial smooth muscle contraction is observed due to the release of histamine, a chemical mediator, from mast cells. The above-mentioned release of histamine from mast cells occurs due to a decrease in C-AMP.
If the content is increased, the above-mentioned histamine release can be suppressed, and as a result, it is effective in treating or preventing allergic asthma. Cyclic Nucleotides in Disease
Baltimore, Univ. Parkpress) p. 211 (1975
As shown in 2010, insulin is the most important hormone that lowers blood sugar. The secretion of insulin is enhanced by glucose, catecholamines, glucagon, etc., and prior to this secretion, an increase in C-AMP occurs. An increase in C-AMP in the pancreas has the effect of promoting insulin secretion. Therefore, the action of PDE inhibitors is effective in increasing C-AMP, promoting insulin secretion, and thereby treating diabetes. Japanese Heat Journal, Volume 16, Page 76 (1975) states that in vascular endothelial cells undergoing arteriosclerosis, the C-AMP content decreases; An increase in fat enhances the lipolytic effect. PDE inhibitors increase C-AMP content,
As a result of promoting lipolysis in vascular endothelial cells, it is useful for preventing arteriosclerosis. As stated in Federation Proceedings, Vol. 30, p. 330 (1971), pharmacological psychotic drugs have a primary site of action, the central nervous system.
-Change AMP content. For example, phenothiazine antipsychotics inhibit activation of PDE by calcium-dependent activator proteins in the brain, increasing intracellular C-AMP concentration,
Furthermore, benzodiazepines, which are tricyclic antidepressants, have a PDE inhibitory effect on the brain, and are therefore effective in treating psychosis. Therefore, PDE
Inhibitors, like these psychotic drugs, can be used to treat psychosis caused by decreased C-AMP levels. Journal of Investment
Dermatology (Journal of Investigative
Dermatology, Vol. 64, p. 124 (1975), psoriasis spreads rapidly in the skin, and in these psoriatic lesions the C-AMP content is reduced;
PDE activity increased approximately three times the normal value, and cyclic guanine monophosphate (C-
GMP) content is increasing. Furthermore, glycogen retention is also observed. C-AMP suppresses cell proliferation and differentiation and promotes glycogenolysis. Papaverine, a clinically known PDE inhibitor, is actually used for the treatment of psoriasis, and it has been found that PDE inhibitors are effective as this therapeutic agent. In the above general formula [], the lower alkyl group includes, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, etc., and the cycloalkyl group includes, for example, cyclopropyl, cyclobutyl, Included are cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Representative compounds of the present invention include the following. 6-[3-(1-methyltetrazol-5-yl)propoxy]carbostyryl, 6-[3-(1-ethyltetrazol-5-yl)propoxy]carbostyryl, 6-[3-(1-propyltetrazole) -5-yl)propoxy]carbostyryl, 6-[3-(1-isobutyltetrazole-5-
yl)propoxy]carbostyryl, 6-[3-(1-cyclopentyltetrazole-)
5-yl)propoxy]carbostyryl, 6-[3-(1-cyclohexyltetrazole-
5-yl)propoxy]carbostyryl, 6-[3-(1-cycloheptyltetrazole-
5-yl)propoxy]carbostyryl, 6-[3-(1-cyclopropyltetrazole-)
5-yl)propoxy]carbostyryl, 6-[3-(1-cyclooctyltetrazole-
5-yl)propoxy]carbostyryl. Compounds of general formula [] can be produced by various methods.
As described in No.-107869 (Japanese Unexamined Patent Publication No. 55-35019), the following reaction formula [In the formula, X represents a halogen atom, and R is the same as above] As shown in the formula, a known 6-hydroxycarbostyryl [] and a tetrazole derivative [] are subjected to a dehydrohalogenation reaction by a conventional method. Manufactured by The compound of general formula [] can be administered to animals and humans as is or together with conventional pharmaceutical carriers. The dosage unit form is not particularly limited and may be appropriately selected and used as required. Examples of such dosage unit forms include oral preparations such as tablets, capsules, granules, and various oral liquid preparations, and parenteral preparations such as injections and suppositories. The amount of the active ingredient to be administered is not particularly limited and can be appropriately selected from a wide range, but in order to achieve the desired effect, it is preferably 0.06 to 10 mg per kg of body weight per day. Further, it is preferable that the dosage unit form contains 1 to 500 mg of the active ingredient. In the present invention, oral preparations such as tablets, capsules, and oral liquid preparations are manufactured according to conventional methods. That is, tablets contain the compound of the present invention, gelatin, starch, lactose,
It is mixed and shaped with pharmaceutical excipients of magnesium stearate, talc, and gum arabic. Capsules are prepared by mixing the compound of the present invention with an active pharmaceutical filler or diluent, and filling the mixture into hard gelatin capsules, soft capsules, and the like. Oral liquid syrups and elixirs are prepared by mixing the compound of the present invention with sweetening agents such as sucrose, preservatives such as methyl- and propylparabens, coloring agents, flavoring agents, and the like. In addition, parenteral preparations are manufactured according to conventional methods, for example, by dissolving the compound of the present invention in a sterilized liquid carrier. The preferred carrier is water or saline. Solvents with the desired clarity, stability and suitability for parenteral use are prepared by dissolving about 1 to 500 mg of the active ingredient in water and an organic solvent and then in polyethylene glycol having a molecular weight of 200 to 5000. Such liquids include sodium carboxymethyl cellulose, methyl cellulose,
A lubricant such as polyvinylpyrrolidone or polyvinyl alcohol is preferably blended. Furthermore, benzyl alcohol, phenol,
A bactericidal agent and a fungicide such as thimerosal, and if necessary, an isotonic agent such as sucrose and sodium chloride, a local anesthetic, a stabilizer, a buffer, and the like may be included. In addition, from the viewpoint of stability, drugs for parenteral administration may be filled into capsules, etc., frozen, water removed using normal freeze-drying techniques, and liquid preparations prepared from the freeze-dried powder immediately before use. can. Furthermore, the phosphodiesterase inhibitor of the present invention has excellent long-lasting action, extremely low toxicity, and has extremely low side effects such as increased heart rate, cardiovascular hypertrophy, and myocardial damage. Next, the present invention will be explained in more detail with reference to production examples, pharmacological tests, acute toxicity data, and formulation examples of the compound of general formula []. Production Example 1 30.6 g of N-γ-chlorobutyrylcyclohexylamine is added to 200 ml of dry benzene, and 36 g of PCl ₅ is added with stirring while cooling the outside with ice and keeping the internal temperature below 20°C. After the addition, the reaction solution was stirred at room temperature for 2 hours, and then concentrated to about half its volume using an evaporator at a bath temperature of 50° C. or lower. 10% in concentrated liquid
140 ml of benzene containing HN ₃ was heated to an internal temperature of 15 ml under stirring.
It takes 90 minutes to drip while keeping the temperature below ℃. After dropping, the reaction solution is left overnight at room temperature. The mixture was refluxed for 3 hours with stirring and then concentrated. Add 200 ml of chloroform to the obtained concentrate for extraction. The chloroform layer is washed with 5% _NaHCO3 water and water and dried _{over Na2SO4} . After removing the desiccant,
The mother liquor was concentrated, and the residue was recrystallized from aqueous isopropanol to give colorless needle-like crystals of 1-cyclohexyl-5.
-28 g of γ-chloropropyltetrazole are obtained.
Melting point: 82-85°C Production Example 2 The following compound was obtained in the same manner as in Production Example 1 above. 1-Ethyl-5-γ-chloropropyltetrazole, colorless liquid, boiling point 160-163°C/2.0mmHg. Production Example 3 3.2 g of 6-hydroxycarbostyryl, 3.3 g of potassium carbonate and 7.7 g of 1-cyclohexyl-5-γ-chloropropyltetrazole are added to 200 ml of dimethylformamide, and the mixture is stirred at 70 to 80°C for 4 hours. After the reaction, dimethylformamide is distilled off under reduced pressure, 300 ml of chloroform is added to the residue for extraction, and the organic layer is washed with dilute NaOH water and water, and dried over Na ₂ SO ₄ . After removing the desiccant and concentrating the mother liquor, the residue was recrystallized from chloroform to give colorless needle crystals of 6-[3-(1-cyclohexyltetrazol-5-yl)propoxy]carbostyryl.
Obtain 3.54g. Melting point: 211-212°C Production Example 4 The following compound was obtained in the same manner as in Production Example 3. 6-[3-(1-ethyltetrazol-5-yl)propoxy]carbostyryl, pale yellow powder crystals, melting point 179-181.5°C 6-[3-(1-cyclopentyltetrazole-)
5-yl)propoxy]carbostyryl, colorless needle crystals, melting point 196.5-197.5℃ 6-[3-(1-cyclooctyltetrazole-)
5-yl)propoxy]carbostyryl, colorless needle crystals, melting point 220-220.5℃ 6-[3-(1-isopropyltetrazole-5)
-yl)propoxy]carbostyril, colorless needle-like crystals, melting point 202-203°C [Pharmacological test 1] Cyclic AMP phosphodiesterase inhibitory effect: The cyclic AMP phosphodiesterase inhibitory effect of the compound of the present invention was evaluated by Biochimica et Biophysica Acta. Biophysica
429, pp. 485-497 (1976) and Biochemical Medicine (1976)
10, pp. 301-311 (1974). That is, the rabbit PRP sample used in Pharmacology Test 1 was further centrifuged at 3000 rpm for 10 minutes, and the platelets were then mixed with Tris-HCl buffer (50 mmol).
A solution of MgCl ₂ (1 mmol) (PH7.4,
10 ml) was added, the platelets were ground using a homogenizer, then frozen and thawed twice, and then sonicated.
The supernatant was subjected to ultracentrifugation and used as a crude enzyme solution. Add 10 ml of this crude enzyme solution to Tris-HCl buffer (PH
The column was passed through a DEAE-cellulose column buffered with 6.0, 50 mmol) and washed and eluted with 30 ml of the same buffer. This was eluted using a sodium acetate-Tris-HCl buffer solution in fractions of 5 ml each at a flow rate of 0.5 ml/min using a linear gradient method (total eluate volume, approximately 300 ml). This results in 100
A fraction with a weak activity of less than 2 nanomoles/ml/min at a high cyclic AMP substrate concentration of micromolar and a strong activity of more than 100 pmol/ml/min at a low cyclic AMP substrate concentration of 0.4 micromolar was obtained. , cycle this
It was used as AMP phosphodiesterase. Tris-HCl buffer (PH8.0,
40 mmol. Bovine serum albumin 50μg and
Mix the substrate solution (containing 4 mmol of _MgCl2 ) with 0.2
ml was prepared. 0.2 ml of the cyclic AMP phosphodiesterase solution was added to the above substrate solution, and the mixture was reacted at 30°C for 20 minutes to convert tritiated cyclic AMP to tritiated 5'-AMP. The reaction solution was immersed in boiling water to stop the reaction, and then cooled in ice water. Add 0.05 ml of snake venom (1 mg/ml) to this and react at 30°C for 10 minutes to generate tritium 5'-
AMP was further changed to tritiated adenosine, and the reaction solution was passed through a cation exchange resin to adsorb tritiated adenosine, washed with distilled water, and eluted with 1.5 ml of 3N aqueous ammonia. The phosphodiesterase activity of this eluate was measured by measuring the tritium adenosine produced using a liquid scintillation counter in a conventional manner. From this result, the phosphodiesterase activity value (Vs) of the test compound at each concentration was determined, and from the activity value [Vs) of the control (water not containing the test compound), the phosphodiesterase inhibition rate (%) was calculated using the following formula. Calculated. Phosphodiesterase inhibition rate (%) =Vc-Vs/Vcx100 The following compounds were used as test compounds. Compound A: 6-[3-(1-cyclohexyltetrazole-
5-yl)propoxy]carbostyryl compound B: 6-[3-(1-isopropyltetrazole-5
-yl)propoxy]carbostyryl compound C: 6-[3-(1-cyclooctyltetrazole-
5-yl)propoxy]carbostyryl As a control, known papaverine and 1
-Methyl-3-isobutylxanthine was also measured in the same manner. The results are shown in Table 1.

[Pharmacology test 2]

Cerebral blood flow increasing effect: The cerebral blood flow increasing effect of the compound of the present invention was investigated in the Journal of Surgical Research.
Research), Vol. 8, No. 10, pp. 475-481 (1968
It was measured according to the method described in 2010). That is, using a mixed dog (male, weight 12-20 kg),
The animal was fixed in a prone position under bentobarbital sodium anesthesia, and forced respiration was performed at 20 ml/Kg and 20 times/min. The skull is exposed and removed with a grinder to expose the superior sagittal sinus, and venous blood is guided externally by cannulation. The amount of blood flowing out was measured using an electromagnetic blood flow meter, and then the number of drops per 10 seconds was measured using a drop counter. The rate of increase was calculated by comparing the number of drops in 30 seconds at the peak of increase before and after drug administration. Drugs were dissolved in dimethylformamide, diluted with saline, and administered via a cannula inserted into the femoral vein. Papaverine was also used as a control drug. The results obtained are shown in Table 2.

[Pharmacology test 3]

Hypotensive effect: The hypotensive effect of the compounds of the present invention was examined by non-invasively measuring systolic blood pressure under anesthesia using the Tail Cuff method. The experiment was conducted using the following two types of rats. 1 Gold blatt direnal hypertensive rats (RHR); Wistar male rats weighing 160 to 180 g were placed under ether anesthesia with a silver clip with an internal diameter of 0.2 mm attached to the left renal artery, and the right kidney was left intact. Leave it as is. Four weeks after surgery, subjects with systolic blood pressure of 150 mmHg or higher were fasted overnight and subjected to experiments. 2 Deoxycorticosterone acetate (DOCA)/salt hypertensive rats (DHR); body weight
The left kidney of a male Wistar rat weighing 150 to 170 g was removed under ether anesthesia, and 10 mg/Kg of DOCA was subcutaneously administered once a week from the first week after surgery, and 1% saline was given as drinking water. Five weeks after surgery, subjects with systolic blood pressure of 150 mmHg or higher were fasted overnight and subjected to experiments. Drugs were administered orally, and blood pressure was measured before and 1, 2, 4, 6, and 8 hours after drug administration.
Blood pressure was measured using a recorder (Reclihoriz type 8S,
San-ei instrument) and electrosthygmomanometer PE-
300 (Narco bio-Systems, houston). The results obtained are shown in Table 3.

[Table] Acute toxicity Test compounds A, B, and C were each orally administered to mice and their acute toxicity (LD ₅₀ ) was measured, and they were all 1000 mg or more. Formulation example 1 Preparation of tablets Amount (g) 6-[3-(1-cyclohexyltetrazole-
5-yl)propoxy]carbostyryl 5 Lactose (Japanese Pharmacopoeia) 50 Cornstarch (Japanese Pharmacopoeia) 25 Crystalline Cellulose (Japanese Pharmacopoeia) 25 Methylcellulose (Japanese Pharmacopoeia) 1.5 Magnesium Stearate (Japanese Pharmacopoeia) Product) 1 The above compound of the present invention, lactose, cornstarch, and crystalline cellulose are thoroughly mixed, granulated with a 5% aqueous solution of methylcellulose, carefully dried through a 200-mesh sieve, and then tableted by a conventional method. Prepare 1000 tablets. Formulation example 2 Preparation of capsules Amount (g) 6-[3-(1-cyclohexyltetrazole-
5-yl)propoxy]carbostyryl 10 Lactose (Japanese Pharmacopoeia) 80 Starch (Japanese Pharmacopoeia) 30 Talc (Japanese Pharmacopoeia) 5 Magnesium stearate (Japanese Pharmacopoeia) 1 Finely powder the above ingredients, After thoroughly stirring the mixture to obtain a homogeneous mixture, it is filled into gelatin capsules for oral administration having the desired dimensions.
Prepare 1000 pieces. Formulation example 3 Preparation of tablets Amount (g) 6-[3-(1-cyclohexyltetrazole-
5-yl)propoxy]carbostyryl 1 Polyethylene glycol (molecular weight: 4000) (Japanese Pharmacopoeia) 0.3 Sodium chloride (Japanese Pharmacopoeia) 0.9 Polyoxyethylene sorbitan monooleate (Japanese Pharmacopoeia) 0.4 Metabisulfite Sodium 0.1 Methyl-paraben (Japanese Pharmacopoeia) 0.18 Propyl-paraben (Japanese Pharmacopoeia) 0.02 Distilled water for injection 100 (ml) Add the above parabens, sodium metabisulfite and sodium chloride at 80°C with stirring. Dissolve in about half the volume of distilled water, cool the solution to 40°C, dissolve the compound of the present invention, polyethylene glycol and polyoxyethylene sorbitan monooleate in the solution, and add distilled water for injection to the solution. The final volume is adjusted to the final volume and sterilized by sterilization using a suitable filter paper to prepare an injection.

Claims (1)

[Claims] 1. General formula [wherein R represents a lower alkyl group or a cycloalkyl group] A phosphodiesterase inhibitor comprising a tetrazolylpropoxycarbostyryl derivative represented by the following as an active ingredient. 2 General formula [In the formula, R represents a lower alkyl group or a cycloalkyl group] A circulation improving agent characterized by containing a tetrazolylpropoxycarbostyryl derivative represented by the following as an active ingredient. 3 General formula [wherein R represents a lower alkyl group or a cycloalkyl group] An antihypertensive agent characterized by containing a tetrazolylpropoxycarbostyryl derivative represented by the following as an active ingredient.