WO1998035673A1

WO1998035673A1 - Drugs for relieving peripheral circulation disorder

Info

Publication number: WO1998035673A1
Application number: PCT/JP1998/000587
Authority: WO
Inventors: Yuji Okada
Original assignee: Kirin Beer Kabushiki Kaisha
Priority date: 1997-02-14
Filing date: 1998-02-13
Publication date: 1998-08-20
Also published as: AU5880198A

Abstract

Drug having a vasodilative effect and being efficacious in preventing and treating peripheral circulation disorder. These drugs contain as the active ingredinet 5-amino-N-cyano-N'-[2-(2-chlorophenyl)ethyl]-3-pyridinecarboximidamide or salts thereof and are useful as drugs having, in addition to the above effect, the effects of relieving hyperinsulinemia, impaired glucose tolerance, insulin resistance and visceral fat accumulation.

Description

Description Peripheral circulatory disorder improver Technical field

The present invention relates to improvement of peripheral circulatory disorders, and more specifically, a peripheral circulatory disorder ameliorating agent, a pharmaceutical agent comprising a specific pyridinyl ruboximidamide compound or a pharmaceutically acceptable salt thereof as an active ingredient. The present invention relates to the use of the above-mentioned compound for the production of a compound, and a therapeutic method using the above-mentioned compound.

Background technology

Peripheral circulatory disorders are caused by stenosis of the blood vessel lumen due to arteriosclerosis, embolism due to atherosclerotic tissue debris, or blood clot formation, vasculitis, contraction of blood vessels or increased blood viscosity, etc. It is a condition in which peripheral tissues fall into an ischemic state. Disorders associated with peripheral circulatory disorders of the extremities include chronic arterial obstruction represented by obstructive arteriosclerosis and obstructive plug vasculitis (Buerger ^), progressive systemic sclerosis, systemic erythematosus, vibration disease, Reino's disease, aneurysm or various vasculitis are known. In the case of limb peripheral circulatory disorders, clinical symptoms include cold and numbness in the limbs, intermittent claudication, resting pain, ulcers and necrosis. Surgical treatment, exercise therapy and drug therapy or a combination thereof are currently being used to improve these symptoms or prevent the symptoms from becoming worse. The drugs used in pharmacotherapy are mainly antiplatelet drugs and vasodilators (pharmacotherapy for chronic arterial occlusive disease, medicine and drugs, Vol. 29, pp. 202-803, 199 3 Year) . At present, there are no drugs that directly improve vascular lesions, but vasodilators can be expected to improve symptoms by increasing blood flow in the collateral circulation based on vasodilation and improving circulation by reducing spasm. Can be expected to prevent the progression of the lesion (selection of treatment for chronic arterial occlusion) Cho, The rapeutic Research, Vol. 17, pages 2528-2532, 1996).

However, not all vasodilators are effective in treating peripheral circulatory disorders or chronic arterial occlusion, and drugs that increase blood flow at ischemic sites are suitable. In addition, vasodilators have a considerable effect on lowering blood pressure, and the degree of blood pressure reduction may decrease the blood flow at the ischemic site. May not be used to treat For example, nifedipine, a calcium antagonist, is ineffective in chronic arterial occlusion (Effects of nifedipine in intermittent claudication, J Vase Med Biol, 2, 94-99, 1990; peripheral arterial occlusion) Effect of nifedipine on patients with sickness, Angio 1 o gy. 34, 46-52, 1983).

Reports of potassium channel openers improve peripheral tissue blood flow and oxygen supply in rat chronic arterial occlusion models (increases in blood flow and oxygen partial pressure with force-ream channel openers in chronic arterial occlusion disease models, N aunyn—Schmi edebergs Arch Pharmacol, 337, pp. 341-346, 1988) and promotes high-energy phosphate recovery in rat arterial occlusion models using high-energy phosphate levels as indicators. Report (Therapeutic effects of force-rheum channel openers in peripheral vascular disease, Cardi 0 V asc Drugs Ther, 7, 555-563, 1993) There are no potassium channel openers used to treat obstruction.

Currently, most vasodilators used in the treatment of chronic arterial occlusion are oral staglandin preparations. However, prostaglandin preparations have an antiplatelet effect, and as a side effect, bleeding tendency is observed, and the combined use with other antiplatelet drugs ゃ anticoagulants is considered difficult. Therefore, it is useful for chronic arterial occlusion, etc. If effective and pure vasodilators are provided to treatment sites, it will be easier to use them in combination with drugs such as antiplatelet drugs and anticoagulants, enabling more effective, less side effects, and pharmacotherapy. . On the other hand, arteriosclerosis, which is a cause of chronic arterial occlusion, is more likely to occur due to hyperlipidemia, hypertension, hyperinsulinemia, impaired glucose tolerance, insulin resistance, visceral fat accumulation, or a combination thereof. << Known (Risk factor accumulation in adult diseases,

Mebio, Volume 13, pages 22-27, 1996). In addition, about 40% of patients with chronic arterial occlusion have diabetes (Therapy for Chronic Arterial Occlusion, Therapeutic Res, 17, 2528-2532, 1996). It is known to have impaired glucose tolerance, insulin resistance, and visceral fat accumulation. Therefore, if there is a drug that has the effect of suppressing the risk factors of arteriosclerosis and the cause of diabetes, it is more effective for preventing chronic arterial occlusion, preventing or treating the progress of the disease state, but such drugs are not known. Not been.

W093Z15057 discloses that 5-amino-N-cyano-N '-[2- (2-chlorophenyl) ethyl] -3-pyridinecarboxyimidamide has a blood pressure lowering effect, Its action is based on the opening of the force channel (KRN4884 relaxation in the coronary vessels of the septum, Genera 1 Pharmaco 1 ogy, 27, 985-989, 1996). Japanese Patent Application Laid-Open No. 08-165239 discloses 1) suppression of cultured vascular smooth muscle cell proliferation, 2) suppression of intimal hyperplasia after endothelial injury of rat carotid artery by balloon catheter, and 3) reduction of serum triglyceride. It is disclosed that it is effective in preventing or treating arterial sclerosis.

There is no specific description about the improvement of arterial occlusion and its risk factors such as hyperinsulinemia, impaired glucose tolerance, insulin resistance, and internal fat accumulation. Disclosure of the invention

The present invention is intended to solve the above-mentioned problems, and an object of the present invention is to improve the symptoms of peripheral circulatory disorders of the limbs by vasodilating action, and to prevent and prevent the progression of peripheral circulatory disorders. The goal is to provide a drug that is both therapeutic and prophylactic.

As a result of intensive studies to achieve the above object, the present inventor has found that a specific pyridine-functional lipoxyimidamide derivative, that is, 5-amino-1-N-cyano— [2- (2-chlorophenyl) ethyl] _3— We found that pyridinecarboxyimidamide had an improving effect in arterial occlusion model rats, and also found an effect of increasing skeletal muscle blood. Therefore, this pyridinecarboximidamide derivative is a drug that improves the symptoms of limb peripheral circulation disorder by vasodilating action. Furthermore, the pyridine carboximidamide derivative has also obtained new findings having an effect of improving hyperinsulinemia, impaired glucose tolerance, insulin resistance, and visceral fat accumulation. The present invention has been completed based on the above findings. That is, the drug according to the present invention comprises the pyridinecarboximidamide compound described above.

(5-amino-N-cyano-N '— [2- (2-chlorophenyl) ethyl) —3 —pyridinecarboxyimidamide) or a salt thereof as an active ingredient In other aspects, it is also a therapeutic or ameliorating agent for hyperinsulinemia, impaired glucose tolerance, insulin resistance or endocrine fat accumulation.

The present invention also relates to the use of the above compound for producing an agent for improving peripheral circulatory disorders or treating arterial occlusion.

Further, the present invention provides a method for improving peripheral circulatory disorder or a method for treating arterial obstruction, which comprises administering an effective amount of the above compound to a patient in need of improving peripheral circulatory disorder or treating arterial obstruction. Also concerns. BEST MODE FOR CARRYING OUT THE INVENTION

Pyridine-powered ruboximidamide compound

The pyridinecarboximidamide compound used in the present invention is represented by the following formula (I): 5-amino-N-cyano-l -'- [2- (2-chlorophenyl) ethyl] -3-1-pyridinecarboximidamide Or a salt thereof, which has an effect of improving peripheral circulatory disorder or hyperinsulinemia, impaired glucose tolerance, insulin resistance, and visceral fat accumulation, as confirmed in the Examples below. It can be used as a prophylactic, ameliorating or therapeutic agent for these disorders.

(I)

The compound represented by the formula (I) as described above can be produced by any suitable method, for example, by the method described in the above-mentioned WO93-15757. Can be manufactured. This method can be summarized as follows.

In this method, an alkoxydionion is allowed to act on a cyanopyridine compound to produce an imidate compound, which is reacted with cyanamide to convert it into Ν-cyano-pyridine carboximide, which is then converted into various amines. This is a method for obtaining the desired (Ν-cyano-N′-substituted) pyridinecarboximimidamide compound by reacting with a compound. The specific method is also described for reference in the preparation examples of Examples below.

The compound represented by the general formula (I) used as an active ingredient in the present invention is a basic compound It has a nitrogen atom and can form acid addition salts. Examples of the acid to form an acid addition salt include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid, or acetic acid, propionic acid, maleic acid, and oleic acid. Organic acids such as lumitic acid, citric acid, succinic acid, tartaric acid, fumaric acid, glutamic acid, tontoic acid, methanesulfonic acid, toluenesulfonic acid, and radialsulfonic acid can be given. When the acid addition salt is used as a medicine, it goes without saying that the acid must be pharmaceutically acceptable.

The drug of the present invention

The pharmaceutical agent according to the present invention contains 5-amino-N-cyano-N '-[2- (2-chlorophenyl) ethyl]-3-pyridinecarboxyimidamide or a salt thereof as an active ingredient. As shown in the examples below, peripheral circulatory circulatory disease reduces peripheral tissue obstruction by reducing tissue damage around the occluded site, and improves these disorders by increasing blood flow at the ischemic site. It can be used as a disorder ameliorating drug or as a cure for arterial obstruction.

In addition, the pharmaceutical agent according to the present invention suppresses the increase in insulin without increasing the blood glucose level and prevents the decrease in insulin sensitivity in hyperinsulinemia and insulin resistance as shown in the Examples below. In addition, hyperinsulinemia improves glucose intolerance by suppressing excessive rise in blood glucose level during Darcos challenge and reducing visceral fat accumulation by lowering fat tissue weight. Can be used as a drug for improving, preventing, or treating sickness, impaired glucose tolerance, insulin resistance, and internalization of fat accumulation.

Therefore, in the present invention, the term “ameliorating agent” or “therapeutic agent” includes any of a preventive agent (agent), an improving agent (agent) and a therapeutic agent (agent).

In view of the above, in the present invention, the improvement of peripheral circulation (or improvement of arterial occlusion) increases the blood flow power of skeletal muscle, and increases glucose and triglyceride intake to skeletal muscle. Increase in the amount of cerebral hypertension, impaired glucose tolerance, insulin resistance, and improvement of visceral fat accumulation. In other aspects, it can be said to be a drug for improving, preventing or treating hyperinsulinemia, impaired glucose tolerance, insulin resistance and visceral fat accumulation. That is, the above-mentioned agent for improving peripheral circulation or therapeutic agent for arterial obstruction according to the present invention also includes an agent for improving, preventing or treating hyperinsulinemia, impaired glucose tolerance, insulin resistance and visceral fat accumulation. You.

The pharmaceutical agent of the present invention can be administered orally or parenterally (intramuscularly, subcutaneously, intravenously, transdermally).

When the pharmaceutical agent of the present invention is administered orally, it is administered as a tablet, granule, powder, capsule or the like, and when administered parenterally, it is injected, suspension, sublingual tablet, suppository, patch , Lotions or ointments. To prepare these preparations, excipients, binders, disintegrants, lubricants, stabilizers and the like can be added. As an excipient, for example, lactose, starch, crystalline cellulose, mannitol, maltose, calcium hydrogen phosphate, light gay anhydride, calcium carbonate, etc., as a binder, for example, starch, polyvinylpyrrolidone, hydroxypropyl cellulose , Ethyl cellulose, carboxymethyl cellulose, gum arabic, etc., as disintegrants, for example, starch, carboxymethyl cellulose, etc., as lubricants, for example, magnesium stearate, talc, hardened oils, etc., as stabilizers, for example, lactose, Mannitol, maltose, polysorbates, polyoxyethylene hard castor oil and the like.

These components can be used to produce tablets, granules, capsules, injections and the like.

When applying the pharmaceutical agent of the present invention to humans, it should be administered intravenously, intramuscularly or orally.

1 Prefer to do more. The method of administration and the dosage or therapeutically effective amount of the agent of the present invention will vary depending on the circumstances of the individual patient to be treated, for example, body weight, sex, susceptibility, time of administration, concomitant agent, patient or degree of the disease. Needless to say, the appropriate dose and the number of doses under certain conditions must be determined by a dosage determination test by a specialist based on the above guidelines, but it is usually oral administration per adult per day The amount is about 0.001 to 1.0 mg mg as an active ingredient, and the intravenous dose is about 0.1 to 100 gZ as an active ingredient.

〔Example〕

Hereinafter, the pharmacological effects and formulation examples of the pyridinecarboximidamide compound used in the present invention will be described.

Test compound

(1) Compound 1 (compound used in the present invention): 5-amino-N-cyano N '— [2- (2-chlorophenyl) ethyl] -13-pyridinecarboximimidamide

(2) Compound 2 (control compound): N-cyano-N '-(2-ditroxityl) — 3-pyridinecarboxyimidamide

Preparation of test compound

The method for producing compound 1 is described, for example, in WO 93-15057, and the method for producing compound 2 is described, for example, in Japanese Patent Publication No. 6-62567.

Hereinafter, a specific method for synthesizing Compound 1 will be described for reference.

(1) Synthesis example of compound 1

First, 5-amino-3-cyanopyridine is prepared.

5-Amino-3-cyanopyridine is a known compound, and is described in, for example, Journal of Medicinal Chemistry, Vol. 10, No. 2, 149-154 (1967). It can be prepared by a method.

Dissolve 5-amino-3-cyanopyridine (380 mg, 3.19 ol) in 1-propanol (15 ml) and blow hydrogen chloride gas at 0-5 ° C for 15 minutes. Stir at room temperature for 22 hours. After the reaction, the solvent is distilled off under reduced pressure, and the concentrate is made basic by adding a saturated aqueous solution of sodium hydrogen carbonate (5 Oml), and then extracted with a chloroform port (50 mIX3). The pore-form layer was dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure, and then subjected to silica gel column chromatography (Wakogel C-200, 25 g). 5-Amino-3-pyridinecarboximide (526 mg, 92% yield) is obtained as a colorless oil.

IR (neat) cm " ¹ 3330, 3200, 1630, 1590, 1080.

^{1 H-NMR (90 MHz,} CDC1 3): 5 (ppm) 8, 37 (1H, br s, H-2), 8.14 (1H, d, J = 2.6 Hz, H-6), I 31 (1H , m, H-4), 4.18 (2H, t, 1-6.6 Hz,

CH ₂ CH ₂ CH ₃ ), 1.11 (2H, m, 0CH ₂ CH ₃ ), 1.03 (3H, t, J-7.0 Hz, OCH CHg),

Subsequently propyl = 5-amino 3_ pyridine carboxyanhydride Consequences Mi de one preparative (11 Omg, 0. 6mmol) was dissolved in Asetonitoriru _{(1 ml), NaH 2 P0} 4 - 2H n 0 (375mg, 2.4m mol), Dissolve Na ₂ HPO ₄ (85 mg, 0.6 bandol) and NH ₂ CN (50 mg, L 2miiiol) in water (2 ml) and stir at room temperature for 19 hours. After the reaction, the reaction mixture was extracted with chloroform (20 ml x 3). Carboxymidate (10 Omg, 82% yield) is obtained as a crude oil.

IR (neat) cm ^-1 3350, 2220, 1610, 1320, 760.

¹ H-NMR (90 MHz, CDCl ^): o (ppm) 8.51 (1H, d, 1 = 2. Q Hz, H-2), 8.27 (1H, d, 1 = 2.6 Hz, H-6), 7.72 (1H, dd, 1-2.fl, 2.6Hz, H-4.39 (2H, t, J = 6.6Hz, QCH ₂ CH ₂ CH ₃ ), 4.12 (2H, brs, NH ₂ ), 1.83 (2H, m, 0CH ₂ CH ₃ ), 1.05 (3H, t,} = 1, 7 Hz, 0CH ₂ CH ₂ CHg).

Propyl = 5-amino-1-N-cyano-3-pyridinecarboximidate (100 mg, 0.49 mmol) was dissolved in methanol (1 ml), and 2- (2-chlorophenyl) ethylamine (155 mg, 1. Ommol) And stir at room temperature for 20 hours. After the reaction, the reaction solution is concentrated under reduced pressure, and the obtained residue is crystallized from getyl ether Z-ethyl acetate to give the title compound (10 Omg, 0.33 ol, yield 68%) as colorless crystals.

p 184 ° C

IR (neat) cm " ¹ 3200, 2 Π 0, 1570.

FD-MS m / z 299 ( M, C 15 H 5 CI).

^{1 H-NMR (90MHz, CD} 3 0D): δ (pm) 8.08 (1H, d, J = 2.6 Hz, H-6), 7.83 (1H, br s, H-2), 1.5-7.2 (5H, H-4, C _g CI), 3.73 (2H, t, J 3 Hz,

NHCH ₂ CH ₂ Cg H ₄ CI), 3.13 (H, t,]-7.3 Hz, NHCH ₀ CH ₂ C _g H ₍ CI).

(2) Preparation of compound 2

Compound 2 can be synthesized by the method described in the above-mentioned JP-B-6-62567 according to the method for synthesizing compound 1 described above.

Test Example 1: Effect on lauric acid-induced arterial occlusion rat (1)

[^ Method] Wi star according to the method of Ashida et al. (Protective effect of chic-mouth vidin on experimental rat peripheral arterial occlusion, Thrombosis Resear ch ^ 18, 55-67, 1980) Lactic acid (1 mg) was injected into the femoral artery of a 10-week-old (Nippon-Chain Rivals, Inc.) rat to cause thrombotic arterial occlusion, and the condition of the hind paws was observed. Compound 1, beraprost sodium as a positive control (therapeutic agent for chronic arterial occlusion) and its vehicle (0.5% Shi methylcellulose solution) twice daily from lauric acid injection 30 ^__ minutes before and 1 to 3 days after, administration of the 4 after 10 days once a day, to 6 m LZk gavage g (the stomach by oral sonde ).

The condition of the hind paws was observed 10 days later, and the degree of lesion of the paws was classified into the following 0 to 5 to determine the effect. The judgment was based on the following evaluation. Score 0: no change, 1: edema or redness of the foot, 2: mummification or necrosis of the nail, 3: mummification or necrosis of the finger, 4: mummification or necrosis up to half of the instep, 5: whole of the instep Mummification or necrosis. The significance test was performed by the U test of Mann-Whitney.

[Results] The results are shown in Table 1.

table 1

Number of drugs Case standard error of score average Comparison with vehicle group Media 7 4.00 Sat I 38

Compound 1 (1 mg / kg) 8 1.00 ± 0.54 ρ <0.05 Compound 1 (3 mg / kg) 11 0.91 ± 0.34 ρ <0.01 Beraprostonadium 10 1.70 ΰ.45 ρ <0.01

(0.3 mg / kg) As shown in Table 1, Compound 1 reduced foot injury due to arterial occlusion Test Example 2: Effect on lauric acid-induced arterial occlusion rat (2)

[Method] Wistar (9-week-old, Japan Charles River, Inc.) Injected lmg of lauric acid into the femoral artery of a rat to induce thrombotic arterial occlusion and observe the condition of the hindpaw . Compound 1, diluvadipine, a calcium antagonist, as a comparator, beraprost sodium (a therapeutic agent for chronic arterial occlusive disease) and a vehicle (0.5% carboxymethylcellulose solution) as a positive control 1 hour before lauric acid injection and After 1 to 10 days, gavage administration of 6mlZkg twice a day (by oral probe) Administration into the stomach).

The condition of the hind paws was observed 10 days later, and the degree of lesion of the paws was classified into the following 0 to 5 to determine the effect (score 0: no change, 1: foot edema or redness, 2 : Mummification or necrosis of the nail; 3: mummification or necrosis of the finger; 4: mummification or necrosis of up to half of the instep; 5: mummification or necrosis of the entire instep. The significance test was performed by the U test of Mann-Whitney.

[Results] The results are shown in Table 2.

Table 2

Number of drug cases Score average soil standard error Comparison with vehicle group Medium 3.55 Sat 0.28

Compound 1 (0.3 mg / kg) 2.55 C 0.41

Compound 1 (1 mg / kg) 2.21 ± 0.41 p <0.05 Compound 1 (3 mg / kg) 2.21 ± 0.41 p <0.05 Nirvadipine (3Gfflg / kg) 3.91 Sat 0.25

Beraprost Natrium 2.73 Sat 0.36

(0.3 mg / kg) As shown in Table 2, Compound 1 reduced foot injury due to arterial occlusion _c Test Example 3: Effect on lauric acid-induced arterial occlusion rat (3)

[Method] Wistar (13-week-old, Japan Charles River, Inc.) Inject 1 mg of lauric acid into the femoral artery of a rat to cause thrombotic arterial occlusion and observe the condition of the hindpaw did. Compound 1, beraprost sodium (a therapeutic agent for chronic arterial occlusion) as a positive control and their medium (0.5% carboxymethylcellulose solution) 6 ml / kg twice a day from 1 day to 3 days after lauric acid injection Was administered by oral gavage (administration into the stomach by oral sonde). The condition of the hind paws was observed 3 days later, and the degree of foot lesions was classified into the following 0 to 5 to determine the effect (score 0: no change, 1: edema or redness of the foot, 2: nail) Mummification or necrosis, 3: mummification or necrosis of fingers, 4: mummification or necrosis of up to half of the instep, 5: mummification or necrosis of the entire instep). The significance test was performed by the Mann-Whitney U test. [Results] The results are shown in Table 3. Table 3 Number of drug cases Score mean standard error Comparison with vehicle group Medium 2.89 ± 0.20

Compound 1 (0.3 oig / kg) 2.22 ± Q.15 P-0.05 Compound 1 (1 mg / kg) 2.00 + P-0.01 Compound 2 (1 mg / kg) 2.11 Sat 3.11 P <0.05 Beraprostonadium 2.33 Sat] Π

As shown in (0.3 mg / kg) Table 3, Compound 1 improved the failure of the foot portion due to arterial occlusion _c

Test Example 4: Effect on lactate-induced arterial occlusion rat

[Method] 0.1 ml of 5% lactic acid was injected into the femoral artery of a Wistar system (13-week-old, Japan SLC, Inc.), thrombotic arterial occlusion was observed, and the condition of the hind legs was observed. Compound 1, Compound 2 and vehicle were administered by gavage at 1 ml before lactate injection (pre-dose) or 3 hours after lactate (post-dose) and twice a day from the following day until the 7th day, 2 ml / kg (oral sonde). Administration into the stomach).

The condition of the hind paws was observed 7 days later, the degree of lesion for each finger was classified into 0 to 4 graduations, and the sum of the grades of the 5 fingers was calculated as the lesion index (grade 0: No change, 1: Blackening is limited to toe, 2: Blackening is limited to finger, Five

3: finger necrosis, 4: finger shedding). The significance test was performed by U-test of Mann-Whitney. Ticlovidine, a drug for treating chronic arterial occlusion, was used as a positive control and administered orally 3 hours before lactate injection.

[Results] The results are shown in Table ⁴ .

Table 4

Drug Dose (mg / kg) Number of subjects Score average soil standard error Comparison with vehicle group Vehicle 11 11.2 Sat 1.1

Compound 1 (pre-dose) 0.5 11 1 3

Compound 1 (pre-administration) 1. 5 11 4.1 Sat 1. 8 Ρ <0.01 Compound 1 (pre-administration) 4.5 10 3.0 0 1.3 3 0.01 0.01 Compound 1 (post-administration) 1.5 5 3.8 Sat 1.5 Ρ <0.01 Compound 1 (post-dose) 4.5 5 1.5 1.5 Sat 0.9 ρ <0.01 Compound 2 (pre-dose) 1. 0 11 5. 2 Sat 1.9 ρ <0.05 Compound 2 (pre-dose) 5.0 119.1 Sat 2.0

Chic Mouth Vidin 300 10 1.6 Soil 0.9 ρ <0.01 As shown in Table 4, Compound 1 and Compound 2 alleviated foot injury due to arterial occlusion. In particular, at higher doses (ί5 mg / kg for compound 1 and 5.0 mg / kg for compound 2), compound 1 was more effective than compound 2. Test Example 5: Effects on adrenergic ergomin-induced arterial occlusion in rats

[^ Method] Wister (13-week-old, Japan SLC Co., Ltd.) 0.1 mg of adrenaline is administered subcutaneously to the tail of the rat in two separate doses, and at the same time 5 mg / kg of ergoquinine subcutaneously to the back Was administered, and thrombotic arterial occlusion occurred, and the condition of the tail was observed. 2 hours before (pre-administration) or 5 hours (post-administration) of Lactate 1 and its vehicle and 2 times daily from the next day to the 13th day by gavage administration of 2 ml Zkg g Administration into the stomach).

The tail condition was observed after 14 days, and the maximum length of the lesion was measured. The significance test was performed by a Student t test. Ticlovidine, a treatment for chronic arterial obstruction, was used as a positive control, and was administered orally 3 hours before adrenaline and ergoymin.

[Results] The results are shown in Table 5.

Table 5

Drug dose (rag / kg) Number of patients Length of lesion () Comparison with vehicle group Vehicle 12.9 +

Compound 1 (pre-administration) 0.5 10.6 g ρ く 0.01 Compound 1 (pre-administration) 1.5 9.1 Sat 13Ρ 0.01 Compound 1 (post-administration) 0.5 9.9 Sat ί 6 p <0.01 Ticlovidine 300 10.4 Sat 0.2 Ρ < 0.01 As shown in Table 5, Compound 1 reduced tail injury due to arterial occlusion. Test Example 6: Effect on lower limb motor dysfunction in arterial occlusion rat

[Method] Wistar rats (10-week-old, Nippon Charls River Co., Ltd.) were ligated just before the bifurcation of the abdominal aorta to the iliac artery, and one week later, the lower limb motor function was measured by forced swimming. . Lower limb motor function was measured by the time the rat was swimming by moving the lower limb in the pool for 5 minutes. Compound 1 or its vehicle was orally administered by gavage (administration into the stomach by oral sonde) at a dose of 0.3 mg / kg twice a day after the arterial ligation for 1 week until the measurement of motor function. The final dose was given 40 to 80 minutes before the motor function measurement. The significance test was performed by a t-test. [Results] The results are shown in Table ⁶ .

Table 6

Drugs Number of swims with lower limbs (minutes)

Simulated cattle rats (normal rats)

Medium 6 1 5 2. 2 ± 20.1 Compound 1 6 1 5 1.7 ± 3 3.5

Arterial occlusion rat

+ Medium 6 99.0 ± 15.9 Compound 16 1 62.5 ± 27.0

+

: P <0.05 compared with sham-operated rat vehicle administration group,

*: P <0.05 Comparison with arterial occlusion rat vehicle administration group

As shown in Table 6 above, the arterial occlusion rat showed a decrease in motor function compared to the normal rat. The administration of Force ^ Compound 1, which was recognized, improved the motor function of the arterial occlusion rat. Compound 1 did not affect the motor function of normal rats.

Test Example 7: Effect on skeletal muscle blood flow

[Method] Wistar (8-11 weeks old, Nippon Charles Riva Co., Ltd.) After ligating the left femoral artery of rats 6-9 weeks later, Compound 1 was intravenously administered under anesthesia. The blood flow of skeletal muscle in the left stomach was measured with a laser Doppler blood flow meter. Blood pressure was also measured at the same time. The significance test was performed by a paired t test. [Results] The results are shown in Table ⁷ c

Table 7

Drug dose (g / kg) No. of patients Blood pressure change (%) Blood flow change (%) Compound 1 1.-4.8 Sat L 3 ** 24.9 ± 15.8 Compound 1 3. -16.4 ± 2.9 93.1 ± 44.0 * Compound 1 10.-31.4 ± 2.mt 140.6 ± 49.4 *

*: P <0.05, **: p * 0.01, ***: ρ * 0.001 Comparison with the value before administration of Compound 1 As shown in Table 7, Compound 1 increases blood at the ischemic site due to arterial occlusion I let it. Test Example 8: Effect on high fructos fed rats

[^ Method] SD rats (8 weeks old) were fed a diet containing 66% fructose for 2 weeks, and administered compound 1 and vehicle for 14 weeks, 2 ml / kg by gavage twice daily.

(Administration into the stomach by oral sonde) was performed.

The high fructose diet was started immediately after administration. Serum glucose and insulin levels and adipose tissue weight around epididymis were measured 4 hours after the last administration of compound 1. The significance test was performed by Dunnett's multiple comparison test or Student's t test.

[Results] The results are shown in Table 8 in / T ヽ

Table 8

Pharmaceutical dosage Number of subjects Glucose--

(mg / kg) (mg / dl) (ϋ / m!) (mg / g) Normal diet

Medium 9 145.2 Sat 7.0 24. is 5.9 4.45 ± 0 32 Fructose diet

Medium 10 165.8 Sat? .8 + 47.2 Sat 6.5 + 4.90 ± 0.25 Compound 1 0.3 9 155.8 ± 4.8 32.5 * 8.1 4.03 ± 0.31 * Compound 1 1.0 10 162.4 Sat 5.1 23.3 * 3.8 * 3.5 is 0.17 ** Compound 1 3.0 9 164.1 * 18 21. 3.1 * 3.69 ± 0.19 **

+: Ρ <0.05 Comparison with normal diet group,

*: Ρ <0.05, **: ρ 0.01 Comparison of fructose-fed rats with vehicle administration group As shown in Table 8, rats with high fructose diet had higher serum insulin levels than rats with normal diet. He presented with hyperinsulinemia. In addition, in the high fructose diet rat, the glucose level was slightly higher than that in the normal diet rat, despite the high insulin level, suggesting a decrease in insulin sensitivity (insulin resistance). Compound 1 was found to be effective in preventing these hyperinsulinemias and insulin resistance. Furthermore, Compound 1 was found to have an effect of reducing the fat weight around the epididymis and suppressing the accumulation of visceral fat. Test Example 9: Effect of high fructose diet rat on glucose tolerance

[Method] SD (8-week-old) rats were fed a diet containing 66% fructose for 2 weeks, and Compound 1 was administered twice daily from the day when the high fructose diet was started until the 14th day. , Performed orally.

Four hours after the administration of compound 1 on day 15, glucose 5 g / kg was orally administered, and the glucose and insulin concentrations in serum immediately before and 15 minutes after the administration were measured. The rats were fasted for at least 18 hours before the measurement. The significance test was performed by the t-test of Student or the test of Aspine-We1ch.

[Results] The results are shown in Table ⁹ .

Table 9

Inverted fiber Glucose (mg / dl) Insulin (U / ml) mm Negative

Normal diet

Rise 10 102.0 ± 2.1 17.9.9 ± 8.7 55.5 Sat 1.2 33.2 2F 4.1 Funorectic meal

Flame ^{9 120. 8 ± 6. 0 + 234.} i ± 5. 7 +++ 18. 4 ± 3. 6 ++ 58. The ^"++ of ^ II 10 113. 8 Sat 4. 1 216. 8 ± 6.5 12.5 Sat 2.5 40.0 ± 6.0 11.0 (rag / kg)

+: P <o. 05, ++: p <o. 01, +++: Fp <o. 001 Normal: ^

*: P <0.05 with fructose-fed rats

As shown in Table 9, rats with a high fructose diet had higher serum glucose levels than normal rats, and abnormal glucose tolerance was observed. It was clarified that administration of Compound 1 lowered serum darcos levels and improved glucose intolerance. Insulin levels were also higher in high fructose diet rats than in normal diet rats, indicating a decrease in insulin sensitivity. Compound 1 suppressed this increase in insulin without increasing the glucose level, indicating that Compound 1 prevented the decrease in insulin sensitivity. As a result, it became clear that Compound 1 has a preventive effect on impaired glucose tolerance and insulin resistance. The preparation examples of the present invention are shown below.

Formulation Example 1

Compound 1 2.Omg

Lactose 80.2mg

Microcrystalline cellulose 31.2 mg

CMC sodium 6.Omg

0.6 mg of magnesium stearate

Total 120.Omg

The above ingredients are mixed and pressurized to make a tablet.

Formulation Example 2

Compound 1 2.0 mg

Lactose 81.8 mg

Corn Nada Flour 55.85 mg

0.35mg magnesium stearate

Total 140.Omg

The above components are mixed and filled into a capsule to form a capsule.

Industrial applicability

As described above, the agent for improving peripheral circulatory disorders according to the present invention has an effect of improving peripheral arterial occlusion and also has an effect of improving hyperinsulinemia, impaired glucose tolerance, insulin resistance, and visceral fat accumulation. It is useful as a drug capable of ameliorating the effects of peripheral circulatory disorders and preventing its progress, that is, a drug having both therapeutic and preventive effects on peripheral circulatory disorders.

Claims

The scope of the claims

1. 5-Amino-N-cyano N '-[2- (2-chlorophenyl) ethyl] — peripheral circulatory disorder characterized by containing 3-pyridinecarboxyimidamide or a salt thereof as an active ingredient Improver.

2. Arterial occlusion characterized by containing 5-amino-N-cyano-N '-[2- (2-chlorophenyl) ethyl] -13-pyridinecarboxyimidamide or a salt thereof as an active ingredient.治治 ■.

3. 5-amino-N-cyano N '-[2- (2-chlorophenyl) ethyl]-3-pyridinecarboxyimidamide or a salt thereof as an active ingredient, An agent for improving hyperinsulinemia, impaired glucose tolerance, insulin resistance or visceral fat accumulation.

4. 5-amino-N-cyano N '-[2- (2-chlorophenyl) ethyl] —Requires an effective amount of 3-pyridinecarboxyimidamide or a salt thereof to improve peripheral circulatory disorders A method for improving peripheral circulatory disorders, which comprises administering to a patient.

5. 5-Amino-N-cyano N '-[2- (2-chlorophenyl) ethyl] —a patient in need of an effective amount of 3-pyridinecarboxyimidamide or a salt thereof to treat arterial occlusion A method for treating arterial occlusion, which is administered to a subject.

6. 5-amino-N-cyano N '-[2- (2-chlorophenyl) ethyl] -3-Use of 3-pyridinecarboxyimidamide or a salt thereof for producing an agent for improving peripheral circulatory disorders.

7. Use of 5-amino-N-cyano-N '-[2- (2-chlorophenyl) ethyl] -3-pyridinecarboxyimidamide or a salt thereof for producing arterial occlusion .