JP6436475B2 - Vasospasm inhibitor - Google Patents

Description

  The present invention relates to an agent for suppressing vasospasm contained in each site of roots, trunks, branches, stems and leaves of plants belonging to the genus Salacia, and more specifically, dry shredding of roots, stems, branches, stems or leaves of plants belonging to the genus Salacia. The present invention relates to a vasospasm inhibitor obtained by extracting water with hot water.

  Myocardial infarction and cerebral infarction are not caused by dysfunction of organs such as the heart and brain itself, but are “vascular diseases” that develop when blood circulation is impaired due to abnormalities in blood vessels distributed there. This total vascular disease is the second leading cause of death in Japan, and according to the 2011 demographic statistics of the Ministry of Health, Labor and Welfare, it accounts for approximately 26% of the causes of death. There are two possible causes of so-called vascular disease. The first etiology is arteriosclerosis, which progresses over time due to risk factors such as high blood pressure and smoking. The second etiology is vasospasm.

The vasoconstriction was dependent on Ca ²⁺ concentration "normal shrinkage", vascular smooth muscle contracts independent of the Ca ²⁺ concentration "vascular abnormality shrinkage (vasospasm)" and is known, blood pressure maintenance, etc. As for the normal contraction responsible for the normal function of the blood vessels, a considerable part has been clarified including its mechanism. For this reason, for diseases such as cerebral infarction, angina pectoris, myocardial infarction and the like, firstly, a therapeutic method and a therapeutic drug by inhibiting normal contraction with obvious mechanism, particularly by inhibiting Ca ²⁺ channel have been reported ( For example, see Patent Document 1).

  However, the above-mentioned vasospasm associated with the onset of cerebral infarction, angina pectoris, myocardial infarction, etc., causing serious symptoms, is considered to occur by a mechanism different from normal contraction, and little details have been clarified. In patients with vasospasm, vascular smooth muscles suddenly contract abnormally without warning, causing severe blood circulation disorders. The existence of risk factors is unclear and suddenly occurs in “healthy” patients, which is feared as the main cause of sudden death (announced by the Japanese Society of Cardiology in 2009), where 60,000 Japanese people die each year . At present, the treatment method for the disease is an apparent treatment method that suppresses the vasoconstriction by suppressing the normal contraction that is less related to the disease. The cause investigation and countermeasures are anxious.

  In recent years, as a composition having an inhibitory action on vasospasm, an inhibitor of cerebral vasospasm comprising an anti-HMGB1 monoclonal antibody as an active ingredient (see, for example, Patent Document 2) or a Rho kinase inhibitor that can be used as an inhibitor of cerebral vasospasm Containing a drug (for example, see Patent Document 3), a preparation for implantation in the brain in which the substance for suppressing cerebral vasospasm is nicardipine, nimodipine or a pharmacologically acceptable salt thereof (for example, see Patent Document 4) In addition, an inhibitor of abnormal vasoconstriction (for example, see Patent Document 5) containing cyberestat as an active ingredient has been reported.

On the other hand, the present inventors have found that sphingosylphosphorylcholine (hereinafter abbreviated as SPC), which is a kind of sphingolipid, is a causative molecule of vasospasm. Furthermore, SPC contracts vascular smooth muscles independently of Ca ²⁺ concentration through activation of Rho kinase, but SPC activates another protein called Fyn, and activated Fyn is on the vascular smooth muscle cell membrane wall. It has been clarified that activating Rho kinase by binding to a structure called membrane raft is an important mechanism in vasospasm (for example, see Non-Patent Documents 1 to 4). Also, a food composition provided for the prevention of vascular disease caused by vasospasm, comprising eicosapentaenoic acid (hereinafter abbreviated as EPA), which is a kind of unsaturated fatty acid obtained from fish and the like, as an active ingredient. (See Patent Document 6).

  On the other hand, the plant of the genus Salacia is known as a vine perennial plant of the asteraceae family that mainly grows in South Asia. These plants of the genus Salacia have been used for diets and the like by drinking roots and stems as tea in the traditional medicine Ayurveda in India and Sri Lanka. Recently, salacinol and kotalanol contained in the root bark of the plant have been used. Has been confirmed to have a potent inhibitory activity against α-glucosidase, and a pharmaceutical composition for suppressing an increase in blood glucose level containing these compounds (see, for example, Patent Document 7) has been reported.

JP 2004-43374 A JP 2007-308436 A JP 2008-069174 A JP 2011-088931 A JP 2008-214309 A JP 2007-112784 A JP 2010-202597 A

Monthly Bioindustry November 2003 "Evasapentaenoic acid (EPA) prevents vasospasm" Nakao F, Kobayashi S. et al. (2002) Circ. Res. 91: 953-960 Shirao S, Kobayashi S. et al. (2002) Circ. Res. 91: 112-119 Somlyo A.V. (2002) Circ.Res. 91: 83-84

  The above-mentioned EPA, which is the only widely used specific ingredient for vasospasm, is difficult to administer oil-soluble fatty acids by injection into the blood due to fish odor, unstable supply due to marine pollution, etc. This is a problem.

  An object of the present invention is to provide a water-soluble vasospasm inhibitor that does not have an unpleasant odor and can be stably supplied.

  The present inventors have paid attention to plant-derived components that have recently attracted attention not only in the agricultural field but also in the medical field. Unlike fish-derived EPA, plant-derived components are easy to control the flavor, can be stably supplied, and water-soluble components obtained by hot water extraction are used as immediate-acting injections. Can stop sudden death.

  The present inventors have targeted vasospasm inhibitors and prepared extracts from various plants and screened them, but the fractions of Salacia oblonga root and hot water extract from the stem are blood vessels. It was confirmed that abnormal contraction (vasospasm) was strongly suppressed and only a slight inhibitory effect was exhibited on normal contraction of blood vessels maintaining blood pressure and blood flow. Furthermore, it has been found that all of the hot water extracts derived from roots, trunks, branches (stems) and leaves have an action of strongly suppressing abnormal vascular contraction (vasospasm). Furthermore, it was confirmed that salacinol and cotaranol, which are contained in plants of the genus Salacia and are known as diabetes-suppressing components, have no inhibitory action on abnormal blood vessel contraction (vasospasm). The present invention has been completed based on these findings.

That is, the present invention is as follows.
[1] It is contained in each part of the roots, stems, branches, stems and leaves of a plant belonging to the genus Salacia, and is obtained by hot water extraction of dried shredded products of the roots, stems, branches, stems or leaves of the plant belonging to the genus Salacia. Vasospasm inhibitor.
[2] The vasospasm inhibitor as described in [1] above, wherein the plant of the genus Salacia is Salacia oblonga.
[3] The vasospasm inhibitor as described in [1] or [2] above, wherein the drying is sun drying.
[4] The hot water extraction is performed by using 1500 to 2500 parts by mass of hot water with respect to 100 parts by mass of the dried shredded product, according to any one of the above [1] to [3] Vasospasm inhibitor.
[5] The vasospasm inhibitor according to any one of [1] to [4] above, wherein the hot water extraction is performed at 85 to 95 ° C. for 2 to 5 hours.
[6] The vasospasm inhibitor according to any one of the above [1] to [5], which is a cerebral vasospasm inhibitor.
In addition, as another aspect of the present invention, [7] a dry shred of roots, trunks, branches, stems or leaves of a plant belonging to the genus Salacia, which is contained in each part of the roots, trunks, branches, stems and leaves of the plant of the genus Salacia A method for suppressing vasospasm, comprising administering an effective amount of a hot water extract of a substance to a subject, and [8] a root, a trunk, a branch of a Salacia plant for use in suppressing vasospasm, A hot water extract of a dried shredded product of roots, stems, branches, stems or leaves of a plant belonging to the genus Salacia, [9] roots, stems, branches, stems and The present invention relates to the use of a hot water extract of a dried shredded product of roots, trunks, branches, stems or leaves of a plant belonging to the genus Salacia, which is contained in each part of a leaf, for the preparation of a vasospasm inhibitor.

  The vasospasm inhibitor of the present invention is highly safe because it can be extracted from edible plants, and exhibits an excellent effect as a vasospasm in that it hardly suppresses normal contractile action. Further, the vasospasm inhibitor of the present invention is highly practical in that it can be used not only for oral administration but also as an immediate-acting injection.

It is a schematic explanatory drawing of (a) suppression of abnormal blood vessel contraction (spasm) and (b) suppression of normal blood vessel contraction depending on the Ca ²⁺ concentration by the Salacia extract. It is a graph which shows the inhibitory effect of the abnormal shrinkage | contraction (vasospasm) and normal contraction of the blood vessel by the hot water extract of all the site | parts of Salacia oblonga root, a trunk, a branch, a stem, and a leaf. The vertical axis shows the inhibition rate. It is a graph which shows the inhibitory effect of the abnormal shrinkage | contraction (vasospasm) of a blood vessel and normal contraction by the stem hot water extract of Salacia oblonga. The vertical axis shows the inhibition rate. The result of having performed the separation process by the column chromatography method of Salacia oblonga root hot water extract is shown.

The vasospasm inhibitor of the present invention is contained in the roots, trunks, branches, stems and leaves of the plants of the genus Salacia, and dried shredded products of the roots, trunks, branches, stems and / or leaves of the plants of the genus Salacia As long as it is a vasospasm inhibitor obtained by hot water extraction, vasospasm means abnormal contraction of vascular smooth muscle that does not depend on cytoplasmic Ca ²⁺ concentration, and depends on the Ca ²⁺ concentration. Shrinkage that does not fall under “normal contraction”.

  The plant belonging to the genus Salacia is not particularly limited as long as it is a plant belonging to the genus Salacia, which naturally grows in Sri Lanka, India and Southeast Asia. Salacia oblonga, S. reticulata, S. reticulata, S. chinensis, S. prinoides, S. latifolia, S. burunoniana, S. grandiflora, S. grandiflora (S. grandiflora) S. macrosperma) can be exemplified, and the above-mentioned Salacia plants can be used singly or in combination of two or more.

  As a method for the drying treatment, the moisture content of the roots, trunks, branches, stems, leaves, or a mixture of two or more of the above-mentioned Salacia plants after harvesting and harvesting is reduced and suitable for storage and transportation. If it is a processing method for preparing a dried Salacia plant, it is not particularly limited, and it is sun-dried by drying the Salacia plant using sunlight; Natural drying to dry the plants; heating and drying by exposing the plants of the genus Salacia under a ventilation of, for example, 40 to 100 ° C., or drying at a room temperature by exposing the plants of the genus Salacia under a normal temperature of ventilation; vacuum drying Or drying by microwave irradiation; etc., and the like, but sun drying is preferable in terms of cost.

  The method for preparing the dried shredded product is not particularly limited as long as it is a method for preparing the dried Salacia plant to a size suitable for hot water extraction. For example, the shredded product is manually shredded or shredded by a grinder. Or by pulverizing with a coarse pulverizer and then finely pulverizing with a fine pulverizer, chopping by passing through a sieve of about 10 mesh after fine pulverization, etc. A method for preparing a dried shredded product of a root, trunk, branch, stem or leaf of a genus plant can be mentioned. Examples of the block piece of the shredded material include 1 to 100 mm, preferably 1 to 10 mm, and particularly 6 mm.

  As a method for performing hot water extraction of the plants of the genus Salacia, the roots and stems of the plants of the genus Salacia are soaked by immersing the dried shredded material of the roots, stems, branches, stems or leaves of the plants of the genus Salacia in hot water. The extraction temperature is not particularly limited as long as it is a method capable of extracting water-soluble components contained in branches, stems, and leaves, and the extraction temperature is 0 ° C. to 100 ° C. at 1 atm or about 1 atm. Preferably, hot water of 60 ° C. to 100 ° C., more preferably 70 ° C. to 98 ° C., more preferably 85 ° C. to 95 ° C. can be mentioned, and under pressure, 100 ° C. or higher, preferably 100 ° C. to 200 ° C. As necessary, stirring treatment and heating reflux can be used in combination.

  The treatment time for performing the hot water extraction is not particularly limited as long as an active ingredient having a vasospasm inhibitory action can be efficiently extracted, and is 30 minutes to 48 hours, preferably 1 to 24 hours, more preferably 10 -20 hours, more preferably 2-5 hours. In addition, the second extraction can be performed using the residue extracted in the first time, and the second liquid of the second time and the first liquid of the first time can be combined into an extraction liquid. In the first time and the second time, 15 minutes to 24 hours, preferably 30 minutes to 12 hours, more preferably 1 to 10 hours, and especially 2 to 5 hours.

  Examples of the extraction water used in the extraction include tap water, well water, distilled water, and deionized water. Distilled water and deionized water are preferable. Moreover, it can replace with water and alcohol aqueous solution can also be used.

  The mass ratio of water (hot water) to the mass of Salacia plant dry shredded product in the water extraction is not particularly limited as long as it is a mass ratio capable of efficiently extracting water-soluble components. For example, dry shredded product 100 Hot water 500 to 5000 parts by mass, preferably 1000 to 3000 parts by mass of hot water, more preferably 1500 to 2500 parts by mass of hot water, and after extraction with hot water, A hot water extract of a Salacia plant can be obtained by removing the hot water extraction insoluble matter by filtration or centrifugation. Further, the Salacia plant hot water extract can be used as a Salacia plant dry extract powder by removing moisture by spray drying or the like.

  The above Salacia plant hot water extract can be used as it is as a vasospasm suppressant, but the extract can be further purified by a suitable purification means such as column chromatography, normal phase chromatography, reverse phase chromatography. It can also be fractionated into a fraction having a high vasospasm inhibitory activity by high performance liquid chromatography, centrifugal liquid chromatography, thin layer chromatography, size exclusion chromatography or the like. A fraction having a high vasospasm inhibitory activity is preferably a fraction having a molecular weight of 25 to 10,000, preferably 50 to 4000, more preferably 100 to 3000, and even more preferably 100 to 2000. can do.

In the present invention, the inhibition rate of abnormal vasoconstriction (vasospasm) can be measured by the same method as the inhibition rate of normal contraction. For example, an arterial mesenteric strip is suspended in an organ chamber in a Magnus tube. After fixing one end and connecting the other end to the transducer, add SPC to the Magnus tube to artificially cause abnormal vasoconstriction (vasospasm), and add another potassium solution to the Magnus tube to remove it. Normal contraction due to polarization (Ca ²⁺ concentration-dependent contraction) is caused, and when the tension becomes plateau and stabilized, each test sample is added to measure abnormal blood vessel contraction (vasospasm) or the rate of suppression of normal contraction. A method can be mentioned.

  As the vasospasm inhibitory ability of the vasospasm inhibitor of the present invention, it is possible to mention that the inhibition rate of the vascular abnormal contraction (vasospasm) is 50% or more, preferably 60% or more, more preferably 70% or more. Preferably, 80% or more is more preferable, 90% or more is particularly preferable, and the inhibition rate of normal contraction of blood vessels is 60% or less, more preferably 50% or less, still more preferably 40% or less, and more preferably 30% or less. Even more preferable, 20% or less is particularly preferable, and 10% or less is very preferable.

  As an ideal form of the vasospasm inhibitor of the present invention, for example, a vasospasm inhibitor having a suppression rate of vascular abnormal contraction (vasospasm) of 75% or more and a suppression rate of normal contraction of 0 can be mentioned, An example of an ideal form of change in tension when a vasospasm inhibitor is added is shown in FIG. FIG. 1 (a) is a graph showing an ideal form of change in tension when a vasospasm inhibitor is administered to a blood vessel that is abnormally contracting (spasm), and FIG. 1 (b) is normal contracting. It is a graph which shows the ideal form of the change of the tension | tensile_strength at the time of administering a vasospasm inhibitor to the blood vessel.

  Since the vasospasm inhibitor of the present invention has a cerebral infarction preventing action, a myocardial infarction preventing action, a cerebral vasospasm inhibiting action after subarachnoid hemorrhage, an angina pectoris preventing action and the like, Produce supplements that prevent or improve vasospasm, as pharmaceuticals such as infarction-related disease treatments, cerebral vasospasm inhibitors, as well as preventive and symptom-improving conditions related to vasospasm Therefore, it can be advantageously used as a pharmacological composition material.

  As the administration form of the vasospasm inhibitor of the present invention, dosage forms such as solutions, emulsions, suspensions and granules are orally administered, and dosage forms such as solutions are administered subcutaneously, intravenously, intramuscularly, and abdominal cavity. Although the form administered parenterally by an internal injection etc. can be mentioned, the administration form which injects a solution etc. intravenously can be illustrated especially suitably.

  The dose of the vasospasm inhibitor of the present invention is not particularly limited as long as it is an amount that can improve a pathological condition involving vasospasm in a subject in need thereof. For example, 1 kg of the above-mentioned dried plant of Salacia plant When a hot water extract extracted with 100 L, preferably 50 L, more preferably 20 L of hot water is administered, 0.1 to 1000 mg / kg body weight, preferably 1 to 100 mg / kg body weight per day However, depending on the symptom, sex, age, etc., the intake can be adjusted as appropriate, and the dose is a dose for preventing the pathological condition involving the vasospasm. Can also be applied for convenience.

  When the vasospasm inhibitor of the present invention is used as a pharmaceutical or a supplement, it is a usual pharmaceutically acceptable carrier, binder, stabilizer, excipient, diluent, pH buffer, disintegrant, solubilizer. In addition, various preparation compounding components such as a solubilizing agent and an isotonic agent can be added, and it can be used in combination with other therapeutic agents for pathological conditions involving vasospasm.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.

[Example 1]
[Preparation of Salacia plant hot water extract]
Salacia oblonga was used as a plant belonging to the genus Salacia. The hot water extract from Salacia oblonga was prepared at both Yamaguchi University and Maruzen Pharmaceutical Co., Ltd. At Yamaguchi University, 2 L of water was added to 200 g of dried shredded products obtained by roughly cutting each part of Salacia oblonga [roots, trunks, branches, stems, leaves] into 6 mm block sections, and extracted at 90 ° C. for 20 hours. At Maruzen Pharmaceutical Co., Ltd., 10 kg of dry shredded material obtained by roughly cutting a Salacia oblonga trunk into 6 mm block slices was added with 10 L of water and extracted at 90 ° C. for 1 hour, followed by filtration to separate the residue from the extract (most solution). The residue was further added with 10 L of water, extracted at 90 ° C. for 1 hour, and filtered to obtain an extract (second solution) that was combined with the previous extract (first solution) to obtain the entire extract.

  Inhibition of vasospasm using Salacia stem hot water extract, Salacia branch and stem hot water extract, Salacia root hot water extract and Salacia leaf hot water extract obtained by hot water extraction at Maruzen Co., Ltd. The experiment was conducted.

[Example 2]
[Vessel spasm suppression experiment]
The vasoconstriction experiment was performed with an 8-channel contraction experiment apparatus using a porcine coronary artery (anterior descending branch). The porcine coronary artery (left anterior descending coronary artery) was cut from the porcine heart processed in the slaughterhouse about 3 cm at a distance of 1 cm from the main trunk branch, and the fat around the blood vessel was removed. The membrane was removed, the endothelium was removed with a cotton swab, and a coronary artery medial strip width 1 mm × length 3 to 4 mm was prepared using a razor.

Krebs solution (123 mM NaCl, 4.7 mM KCl, 15.5 mM NaHCO ₃ , 1.2 mM MgCl ₂ , 1.25 mM CaCl ₂ , 11.5 mM D− having a composition similar to the ionic composition of the extracellular fluid in the Magnus tube the porcine coronary artery mesenteric strip is suspended in a 5 mL organ chamber and fixed at one end, and the other end is connected to a transducer (tension detector) (TB-612T, manufactured by Nihon Kohden Co., Ltd.) Then, isometric tension was measured and the blood vessel tension was evaluated.

Add 30 μM SPC to the Magnus tube to cause abnormal contraction (spasm), add 40 mM high potassium solution to another Magnus tube to cause normal contraction (Ca ²⁺ concentration dependent contraction) by depolarization, and tension When the plateau became plateau and stabilized, each test sample was added to a Magnus tube to which SPC was added and a Magnus tube to which a potassium solution was added so as to have a final concentration of 0.5 mg / mL. Compared. The results are shown in FIG.

(result)
As is clear from FIG. 2, the effect of suppressing abnormal blood vessel contraction was confirmed in the hot water extract of all parts of the roots, trunks, branches, stems, and leaves of Salacia oblonga. The hot water extract of Salacia oblonga stems, branches, and stems has a higher abnormal shrinkage suppression effect than the root hot water extract, and the normal shrinkage suppression effect is equivalent to the root hot water extract. The following was shown. For the hot water extract of Salacia oblonga leaves, the abnormal shrinkage inhibitory effect was slightly lower than the hot water extract of other sites, but it showed a very low inhibitory effect on normal shrinkage. Salacia leaf extract is judged to contain a substance that suppresses abnormal shrinkage and does not affect normal shrinkage, but leaves of plants belonging to the genus Salacia are secured with a certain quality. Therefore, the experiment was continued using a Salacia trunk hot water extract that was easy to secure the raw material and showed a relatively excellent value.

[Example 3]
[Vessel spasm suppression experiment]
Maruzen Pharmaceutical Co., Ltd. was entrusted with hot water extraction treatment from the dried shredded material of Salacia oblonga stem and separation treatment using a column by the column chromatography method of the Salacia stem hot water extract. To an open column packed with Diaion HP20 (Mitsubishi Chemical Co., Ltd.), a sample prepared by dissolving and preparing Salacia stem hot water extract with ultrapure water is added, and the liquid passing / water washing part, 30% ethanol elution part, 50% Separated into an ethanol elution part and a 100% ethanol elution part. A hydrophilic vinyl polymer Toyopearl HW-40C (manufactured by Tosoh Corporation) capable of further molecular weight fractionation was used to separate the liquid passage / water washing portion into fractions 1 to 6. FIG. 3 shows the result of the same vasoconstriction experiment as in Example 2 described above, using each fraction of the obtained Salacia trunk hot water extract.

(result)
Fractions 1 to 3 showed a high inhibitory effect on abnormal contraction and a very low inhibitory effect on normal contraction compared to other fractions (fractions 4 to 6). These fractions mainly contain a compound having a molecular weight of about 150 to 2,000, have a higher polarity than other fractions, and are water-soluble fractions. Therefore, it can be said that there is a high possibility that the active ingredient is also water-soluble. Although the ethanol elution part showed a high inhibitory effect on abnormal contraction, it also showed a high inhibitory effect on normal contraction depending on the ethanol concentration.

[Example 4]
[Vessel spasm suppression experiment]
Separation treatment of the Salacia oblonga root hot water extract prepared in Example 1 using a column chromatography method was performed at Yamaguchi University. To an open column packed with Diaion HP20 (Mitsubishi Chemical Corporation), add 30 mL of the sample with the extract adjusted to about 100 mg / mL with ultrapure water, and separate the water content (fractions 1 and 2 from the difference in color tone). And methanol fraction (separated into fractions 3, 4 and 5 due to the difference in color tone.) Using the obtained fractions of Salacia root hot water extract, the same vasoconstriction experiment as in Example 2 was performed. went.

(Results) Fractions 1 to 5 all showed a high inhibitory effect on abnormal contraction. Among them, fraction 2 did not affect the normal contraction at all and showed a strong effect of suppressing abnormal contraction. Since fraction 2 is a water-soluble fraction, it can be said that there is a high possibility that the active ingredient is also water-soluble. The experimental results of fraction 2 are shown in FIG.

  The vasospasm inhibitor of the present invention can be administered by intravenous injection, and is useful in the field of pharmaceuticals as a therapeutic or prophylactic agent having an immediate effect on serious vascular diseases such as cerebral vasospasm. In addition to contributing to a healthy life, it can be expected to contribute to reducing medical costs in an aging society.

Claims (6)

Vasospasm inhibitor comprising as an active ingredient a hot water extract of Sa roots of Rashia genus plant stem, branches, stems or dry shred the leaves. The vasospasm inhibitor according to claim 1, wherein the plant of the genus Salacia is Salacia oblonga. The vasospasm inhibitor according to claim 1 or 2, wherein the drying is sun drying. The vasospasm inhibitor according to any one of claims 1 to 3, wherein the hot water extraction is performed by using 1500 to 2500 parts by mass of hot water with respect to 100 parts by mass of the dried shredded product. The vasospasm inhibitor according to any one of claims 1 to 4, wherein the hot water extraction is performed at 85 to 95 ° C for 2 to 5 hours. It is a cerebral vasospasm inhibitor, The vasospasm inhibitor in any one of Claims 1-5 characterized by the above-mentioned.