JPH08165237A - Agent for improving blood flowability - Google Patents

Abstract

PURPOSE: To provide an agent for improving blood flowability useful for preventing and/or curing diseases with pathologically reduced blood flowability, having an activity for improving deformation performance of red cells and capable of improving blood flowabilit in the microcircular region. CONSTITUTION: This blood flowability-improving agent contains an isoflavon, preferably daidzein or orobol of the formula (R<1> is OH; R<2> is H or OH; R<3> is OH or a protected OH; R<4> is H or OH) as an effective ingredient and is a curative agent for preventing and/or curing diseases accompanied with a reduction of blood flowability. The effective ingredient compound is a usual compound known as isoflavon or easily available from known raw material compounds or plant raw materials by usual methods. E.g. daidzein is in wide varieties of plants, included in soy beans, Puerariae Radix, etc., as a glycoside (daidzin) in a large amount and easily available. A dose per oral of the agent is 0.02-10mg/kg per adult per day, preferably 0.02-10mg/kg per adult per day as an effective amount.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the invention of a pharmaceutical useful for the prevention and / or treatment of diseases in which blood fluidity is reduced. More specifically, the present invention has the effect of improving the fluidity of blood in the microcirculation region by improving the deformability of red blood cells, and prevents and / or prevents diseases in which blood fluidity is impaired as a pathological condition. The present invention relates to an invention of a medicine useful for treatment.

[0002]

2. Description of the Related Art Heretofore, it has been pointed out that the involvement of hemodynamic factors is involved in the occurrence of an early lesion of arteriosclerosis which frequently occurs in a specific site of the vascular system. Vascular endothelial cells are present in a single layer in the lumen of blood vessels and are constantly exposed to the hemodynamic effects of blood flow. On the other hand, vascular endothelial cells are deeply involved in the regulation of blood vessel growth and tonus through the production of various physiologically active substances.

It is known that atheroma, which is characterized by subcutaneous lipid deposition and proliferation of smooth muscle cells, is likely to occur in a portion such as a branch or a curved portion where the geometrical shape of the blood vessel changes rapidly. ing. Blood flow is changing in such a part, stagnation and turbulence of blood flow occur, and wall shear stress (wa
ll shear stress) intensity and direction change with time.

The wall shear stress is defined by the product of the blood viscosity and the velocity gradient of the blood flow that maximizes the center of the blood vessel. Blood viscosity is governed by various factors, the main ones of which include hematocrit, plasma composition and viscosity, red blood cell deformability, red blood cell aggregate formation, blood coagulation, and red blood cell and platelet aggregation. be able to. By changing these factors, it may be possible to improve blood flow at vascular bifurcations, reduce pressure at stenotic sites in blood vessels, and improve blood fluidity at peripheral circulation sites such as cerebral blood vessels. There is.

Conventionally, pentoxifylline (3,7-dihydro-3,3) has been used as a drug having an effect of improving the deformability of red blood cells.
7-dimethyl-1- (4-acetylbutyl) -1H-purine-2,6-dione:
Sold by Hoechst Japan KK as "Trental") and sarpogrelate ((±) -1- [o-
[2- (m-methoxyphenyl) ethyl] phenoxy] -3- (dimethylamin
o) -2-propyl hydrogen succinate hydrochloride: "Amprag" sold by Tokyo Tanabe Seiyaku Co., Ltd.) is clinically used. Further, vinpocetine, nicergoline, cinepazide maleate, trapidil, dilazeb hydrochloride, flunarizine hydrochloride, cinnarizine and the like have been reported to have erythrocyte deformability.

Of these, pentoxifylline is a drug that improves red blood cell fluidity in the microcirculation region by improving red blood cell deformability and improves blood fluidity. Sarpogrelate is a serotonin 5HT ₂ receptor. It is known as an antagonist to erythrocytes and has a tens of times more potent erythrocyte deformability-improving effect than pentoxifylline. Pentoxifylline is used for the improvement of sequelae due to cerebral thrombosis, and sarpogrelate is used for ischemic symptoms.

[0007]

SUMMARY OF THE INVENTION An object of the present invention is to provide a pharmaceutical useful for the prevention and / or treatment of diseases in which blood fluidity is reduced. From another perspective, the present invention is
It is an object of the present invention to provide a drug having an action of improving the deformability of red blood cells. More specifically, the object of the present invention is to improve the deformability of erythrocytes, and by improving the fluidity of blood in the microcirculation region,
It is an object of the present invention to provide a pharmaceutical which enables the prevention and / or treatment of a disease whose pathological condition is a decrease in blood fluidity.

[0008]

As a result of diligent efforts to solve the above problems, the present inventor has found that daidzein (daidzei)
It was found that isoflavones represented by n) significantly improve the deformability of red blood cells. Further, they have found that these compounds improve blood fluidity, particularly blood fluidity at microcirculation sites, and are therefore useful for the prevention and / or treatment of diseases in which blood fluidity decreases, and complete the present invention. Came to.

That is, the present invention has the following formula:

Embedded image (Wherein R ¹ represents a hydroxyl group, R ² represents a hydrogen atom or a hydroxyl group, R ³ represents a hydroxyl group or a protected hydroxyl group, and R ⁴ represents a hydrogen atom or a hydroxyl group). The present invention provides a preventive and / or therapeutic agent for diseases in which blood fluidity decreases.

According to a preferred embodiment of the present invention, R ¹ is a hydroxyl group, R ² is a hydrogen atom, R ³ is a hydroxyl group, and R ^{4 is}
The preventive and / or therapeutic agent containing a compound in which is a hydrogen atom as an active ingredient; the preventive and / or therapeutic agent containing daidzein or oloball as an active ingredient; the preventive and / or therapeutic agent for improving red blood cell deformability; and The above prevention and / or the disease in which blood fluidity is reduced is a sequelae of cerebral thrombosis
Alternatively, a therapeutic agent is provided.

In the above general formula, R ¹ represents a hydroxyl group and R ² represents a hydrogen atom or a hydroxyl group. When R ² represents a hydrogen atom,
The hydroxyl group of R ¹ may be substituted at any of the 2′-position, the 3′-position, or the 4′-position on the phenyl ring that substitutes at the 3-position of 4H-1-benzopyran-4-one. Good. When R ² represents a hydroxyl group, the two hydroxyl groups are 2'-position and 3'-position, 2'-position and 4'-
Rank, 2'- and 5'-, 2'- and 6'-, 3'- and 4'-
It may be substituted at the position, or at any of the 3'-position and the 5'-position. Of these, R ¹ is preferably a hydroxyl group and R ² is preferably a hydrogen atom, R ¹ represents a hydroxyl group at the 4′-position,
It is particularly preferred that R ² is a hydrogen atom.

In the above general formula, R ³ represents a hydroxyl group or a protected hydroxyl group, and R ⁴ represents a hydrogen atom or a hydroxyl group. When R ⁴ represents a hydrogen atom, R ³ may be substituted at any of 5-position, 6-position, 7-position or 8-position of 4H-1-benzopyran-4-one. When R ³ represents a protected hydroxyl group and R ⁴ represents a hydroxyl group, the substitution positions of R ³ and R ⁴ are 5-position and 6-position, respectively.
5th and 7th, 5th and 8th, 6th and 5th, 6th and 7th
Positions, 6-position and 8-position, 7-position and 5-position, 7-position and 6-position, 7-position and 8-position
It may be any of the positions, 8-position and 5-position, 8-position and 6-position, or 8-position and 7-position. When R ³ and R ⁴ both represent a hydroxyl group, 5-
5th and 7th, 5th and 8th, 6th and 7th, 6th and 8th, or 7th and 8th . Among these, it is preferable that R ³ is hydroxyl R ⁴ is a hydrogen atom, and particularly preferably R ³ is 7-position hydroxyl group R ⁴ is a hydrogen atom.

When R ³ represents a protected hydroxyl group, examples of the hydroxyl protecting group include an alkyl group such as a methyl group, an acyl group such as an acetyl group, a carbonyloxy group such as a t-butoxycarbonyl group, and a trimethylsilyl group. And alkylsilyl groups. However, the protective group for the hydroxyl group is not limited to these,
Any available to those skilled in the art may be used. Further, as such a protective group, those which are rapidly hydrolyzed in vivo after administration to give a compound in which R ³ is a hydroxyl group are preferable. The compound having a protected hydroxyl group may be useful as a so-called prodrug, although it may be useful as an active ingredient of the drug of the present invention.

For example, a sugar such as glucose can be used as the protecting group. For example, a compound such as glucoside, galactoside, or rhamnoside in which a sugar is added to the 7-position or 5-position hydroxyl group may be used as an active ingredient of the medicament of the present invention. Such compounds are well known to those skilled in the art as glycosides of isoflavone compounds. Since such a compound exhibits extremely high water solubility, it is possible to easily produce an injection drug and the like, and R ³ gives a compound having a hydroxyl group by being rapidly hydrolyzed after being administered in vivo. It is a preferred compound as an active ingredient of the medicament of the invention.

A particularly preferred compound as an active ingredient of the medicament of the present invention is daidzein: 7-hydroxy-
3- (4-hydroxyphenyl) -4H-1-benzopyran-4-one; R ¹ : 4'-
OH; R ² : H; R ³ : 7-OH; R ⁴ : H), daidzein 7-glucoside daidzin, orobol
5,7-dihydroxy-3- (3,4-dihydroxyphenyl) -4H-1-benzopy
ran-4-one; R ¹ : 4'-OH; R ² : 3'-OH; R ³ : 7-OH; R ⁴ : 5-O
H) and the like.

However, the active ingredient of the medicament of the present invention is not limited to the above-mentioned preferred compounds. For example, the compounds included in the above general formula can be obtained by the test methods described in the following examples. It is possible to appropriately select the active ingredient according to the above. The compound of the active ingredient of the present invention is a compound known per se as isoflavone, or can be easily obtained from a known raw material compound or plant raw material by a method well known to those skilled in the art. For example, daidzein is widely present in plants and the like, and is contained in soybeans, kudzu roots, and the like in a large amount as a glycoside (daidzin), so that daidzein is easily available.

As the medicament of the present invention, one or more of the above compounds which are active ingredients may be directly administered to a patient, but a pharmaceutical composition containing one or more of the above compounds as an active ingredient. It is preferable that the product is manufactured using a formulation additive and is administered to a patient in the form of a pharmaceutical composition. As the active ingredient of the medicament of the present invention, in addition to the free form compounds included in the above general formula, salts such as acid addition salts and base addition salts, and any hydrates thereof may be used. The dosage form of the medicament of the present invention is not particularly limited, and it can be administered orally or parenterally. Examples of pharmacologically and pharmaceutically acceptable additives used in the production of the above-mentioned pharmaceutical composition include, for example, excipients, disintegrating agents or disintegrating aids, binders, lubricants, coating agents, dyes, Diluent,
Base, solubilizer or solubilizer, isotonicity agent, pH adjuster,
A stabilizer, a propellant, an adhesive, etc. can be mentioned.

Examples of formulations suitable for oral administration include tablets, capsules, powders, fine granules, granules, liquids,
Alternatively, a syrup and the like can be mentioned. Further, as a formulation suitable for parenteral administration, for example, injections, drops,
Examples thereof include suppositories, inhalants, patches and the like. Pharmaceutical preparations suitable for oral administration include pharmacologically and pharmaceutically acceptable additives such as excipients such as glucose, lactose, D-mannitol, starch or crystalline cellulose; carboxymethyl cellulose, starch, or Disintegrators or disintegration aids such as carboxymethylcellulose calcium; binders such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, or gelatin; lubricants such as magnesium stearate or talc; hydroxypropylmethylcellulose, sucrose, polyethylene glycol or Coating agents such as titanium oxide; petrolatum, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water,
Alternatively, a base such as hard fat can be used.

Formulations suitable for transdermal or transmucosal administration include, for example, propellants such as Freon, diethyl ether, or compressed gas; adhesives such as sodium polyacrylate, polyvinyl alcohol, methylcellulose, polyisobutylene, polybutene; cotton cloth. Alternatively, it can be produced by using a pharmaceutical additive such as a base cloth such as a plastic sheet. Formulations suitable for injection or infusion include, for example, distilled water for injection, physiological saline, and solubilizers or solubilizers that can compose aqueous or injection-soluble injections such as propylene glycol; glucose, sodium chloride, D-mannitol. Isotonic agents such as glycerin; inorganic acids, organic acids, inorganic bases or organic bases
It may be produced by adding a pharmaceutical additive such as a pH adjuster.

The medicament of the present invention has an effect of improving the deformability of red blood cells and improves the fluidity of blood in the microcirculation region, and therefore prevents various diseases in which blood fluidity is impaired as a pathological condition. And / or is useful in therapy. Examples of such diseases include peripheral circulatory insufficiency, arteriosclerosis, diabetes, sequelae of cerebral thrombosis, ischemic symptoms during cold sensation, etc. It is not limited to one. Further, it may be used in combination with other drugs used for treatment or prevention of the above diseases.

The dose of the drug of the present invention is not particularly limited,
It may be appropriately selected depending on the administration form, age, body weight, symptoms and the like. For example, in the case of oral administration, per adult day,
0.02-10 mg / Kg, preferably 0.05
Approximately 1.0 mg / Kg may be administered. The administration may be performed once or divided into several times per day, and the administration period can be arbitrarily determined according to age, symptoms and the like. The acute toxicity of daidzein, which is an example of the active ingredient of the medicine of the present invention, is
It is 1,000 mg / kg or more (mouse, po), and it is a compound with extremely low toxicity.

[0022]

【Example】

1. Method for measuring the speed of blood passing through the membrane For in vitro, add 10 μl of a sample dissolved in dimethylformamide to 990 μl of blood, and add at room temperature (2
After incubating at 3 ° C, the raid (Reid, HL,
et al., J. Clin. Path., 29, pp.855-856, 1976), after passing the filter at a constant pressure (ΔP: 200 mm H ₂ O) for 10 seconds, Except
The blood that had passed through the membrane was diluted with 50 ml of water to hemolyze it.
After thoroughly stirring this sample, the absorbance at a wavelength of 506 nm was measured to quantify hemoglobin. In the case of exo-vivo, the blood was left at room temperature for 30 minutes and then directly filtered. All measurements were performed within 6 hours after blood collection. Blood was collected from humans (healthy boys over 20 years old) or rats using 0.1% EDTA, and Nucleopore Hemafil membrane (pore size) was used as a filter.
4.7 to 5.0 μm, diameter 13 mm) was used.

2. Experimental Results (1) In Vitro Membrane-Passing Ability of Blood In order to evaluate blood fluidity, the blood passage rate through the membrane was measured.
As a drug of the present invention, using daidzein and oloball, pentoxifylline reported as a drug for improving erythrocyte deformability as a control (Oguro Masao et al., Basic and Clinical, 13 (6), pp.2033-2038, 1979. ) And a comparative test. The results are shown in Table 1. The rat blood passage rate is 11.6
The concentration was increased by about 9% by μM daidzein and about 10% by 360 μM pentoxifylline, which indicates that daidzein has about 30 times the activity of pentoxifylline. Also, the concentration dependence of daidzein,
Table 2 shows the results of comparison of the effects on human and rat blood. Concentration-dependent increase in blood transmembrane velocity was up to 15.7 μM in human blood and 31.5 μM in rat, which was 1.2 to 1.3 times that of the control. No further increase in passage rate was observed at higher concentrations.

[0024]

[Table 1] ────────────────────────────────── Sample name Absorbance (AU) Ratio control (%) ─ ───────────────────────────────── Control (n = 5) 0.411 ± 0.025 − Daidzein 3.9 μM (n = 5) ) 0.406 ± 0.020 98.8 〃 5.8 μM (n = 5) 0.427 ± 0.023 103.9 〃 11.6 μM (n = 5) 0.447 ± 0.043 108.8 Oroball 5.7 μM (n = 5) 0.453 ± 0.008 109.2 Pentoxy 180 μM (n = 5) 0.418 ± 0.026 101.7 Philin 360 μM (n = 5) 0.450 ± 0.026 109.5 〃 720 μM (n = 5) 0.457 ± 0.045 111.2 ─────────────────────── ───────────

[0025]

[Table 2] ───────────────────────────────── Sample name Absorbance (AU) Ratio control (%) ── ─────────────────────────────── Control 0.347 ± 0.014 − Human blood 3.9 μM 0.360 ± 0.019 103.7 〃 7.9 μM 0.392 ± 0.036 113.0 〃 15.7 μM 0.438 ± 0.016 126.2 〃 31.5 μM 0.430 ± 0.012 123.9 〃 63.0 μM 0.443 ± 0.010 127.7 ──────────────────────────── ─────Control 0.370 ± 0.021 − Rat blood 3.9 μM 0.372 ± 0.010 100.5 〃 7.9 μM 0.400 ± 0.013 108.1 〃 15.7 μM 0.433 ± 0.013 117.0 〃 31.5 μM 0.452 ± 0.018 122.2 〃 63.0 μM 0.450 ± 0.016 121.6 ──── ─────────────────────────────

(2) Exo-vivo blood transmembrane ability Rats fasted for 18 hours (std: Wister / ST strain 10-week-old male, 303-337 g, purchased from Japan LSC Co., Ltd.) 0 .
A sample suspended in 5% CMC was orally administered. Pentobarbital sodium from Dynabot Co., Ltd. 3 hours later
Blood was collected under anesthesia with 50 mg / ml, the blood was stored in ice, and then left at room temperature for 30 minutes to measure the blood membrane passage rate. Five rats were used for each concentration, and one rat was measured twice. Administration of 0.4-10 mg / kg daidzein showed a dose-dependent increase in passage rate, 30 mg / kg
An increase of about 30% was observed in the treated group. The results are shown in Table 3.
Shown in

[0027]

[Table 3] ───────────────────────────────── Sample name Absorbance (AU) Ratio control (%) ── ─────────────────────────────── Control 0.187 ± 0.008 − Daidzein 0.4 mg / kg 0.195 ± 0.017 104.2 〃 2.0 mg / kg 0.230 ± 0.015 123.0 〃 10.0 mg / kg 0.249 ± 0.009 133.2 ──────────────────────────────────

10 mg / kg of daidzein suspended in CMC was orally administered to rats, and blood was collected under anesthesia at 1, 2, and 3 hours after anesthesia, respectively. It examined (it measured twice about 4 each). As a control, a CMC solution was administered, and blood was collected and measured in the same manner (each three mice were measured twice). The results of absorbance (AU) are shown in Table 4. The 0-hour data was obtained by collecting blood under anesthesia without drug administration and measuring it in the same manner. In the control, no significant change in passage speed was observed until after 3 hours, but in the daidzein administration group, a significant increase (about 20%) was observed after 2 hours.

[0029]

[Table 4] ────────────────────────────────── Elapsed time after administration (hours) Control daidzein ─── ─────────────────────────────── 0 0.326 ± 0.028 1.0 0.329 ± 0.021 0.339 ± 0.013 2.0 0.312 ± 0.022 0.383 ± 0.011 3.0 0.327 ± 0.023 0.400 ± 0.014 ──────────────────────────────────

[0030]

The drug of the present invention has an action of improving the deformability of red blood cells, and can improve the fluidity of blood in the microcirculation region, and therefore is a disease in which blood fluidity is reduced as a pathological condition. It is useful for the prevention and / or treatment of

Claims (5)

[Claims] 1. The following formula: (Wherein R ¹ represents a hydroxyl group, R ² represents a hydrogen atom or a hydroxyl group, R ³ represents a hydroxyl group or a protected hydroxyl group, and R ⁴ represents a hydrogen atom or a hydroxyl group). As a preventive and / or therapeutic agent for diseases in which blood fluidity is reduced. 2. The preventive and / or preventive agent according to claim 1, which comprises a compound in which R ¹ is a hydroxyl group, R ² is a hydrogen atom, R ³ is a hydroxyl group and R ⁴ is a hydrogen atom as an active ingredient. Therapeutic agent. 3. The preventive and / or therapeutic agent according to claim 2, which contains daidzein or oloball as an active ingredient. 4. The preventive and / or therapeutic agent according to any one of claims 1 to 3, which improves red blood cell deformability. 5. The preventive and / or therapeutic agent according to any one of claims 1 to 4, wherein the disease in which blood fluidity is reduced is a sequelae of cerebral thrombosis.