JP2633609B2 - Ischemic disease therapeutic agent - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial applications]

TECHNICAL FIELD The present invention relates to a therapeutic agent for ischemic disease comprising a diterpene component tanshinone VI (Tanshinone VI) in a Chinese medicine “Dansan” as an active ingredient.

[Prior art]

(Background) Ischemic diseases such as angina and myocardial infarction are mainly caused by impaired blood circulation to the heart muscle due to stenosis of the coronary arteries.
It is important in fatal adult diseases along with cancer and heart failure. However, cholesterol-lowering drugs and vasodilators have been mainly used as drugs which have been used for the treatment of this disease so far, and drugs intended to suppress myocardial cell membrane damage and protect myocardium during ischemia have not been developed. In recent years, the effects of ancient Chinese and Chinese medicines have been reviewed, and among plant-derived Chinese medicines, recovery of cardiac function and subjective symptoms (dizziness, lightheadedness, cold limbs, etc.) based on remission of angina and dilation of coronary arteries ) Have been reported to be effective, but their pathophysiological and pharmacological evaluations are poor, and especially those active substances against ischemic diseases such as myocardial infarction and angina pectoris. Research on none is even. By the way, Chinese medicine "Dan ginseng" (or purple ginseng) is a Labiatae (Labi
atae), a dried product of the root of the plant Salvia miltiorrhiza Bge., which has been used as a physiologic agent (blood circulation improver) in Chinese medicine for the treatment of angina pectoris and myocardial infarction. Previously unknown. Therefore, as a result of the research, the present inventors previously invented Japanese Patent Application No. 62-102583, and obtained a phenanthrenequine compound, tanshinone I (Tanshinone I) (following formula), isolated from Dansan. Results of ischemia heart disease pathological model search, promoted recovery of myocardial contractility during renormalization, dilation of coronary arteries and ATP
It has been found that it has a significant pharmacological action against ischemic diseases such as inhibition of metabolite release, and that other isolated components are ineffective or have rather toxic effects such as coronary artery contraction.

However, as a result of the subsequent research, a novel phenanthrene compound having a novel pharmacological action on ischemic diseases, tanshinone V ( Tanshinone V) and Tanshinone VI (Tanshinone VI) (following formula) were obtained, and the fact that the latter, tanshinone VI, in particular, has a remarkable anti-ischemic effect was obtained.

[Problems to be solved by the invention]

Therefore, an object of the present invention is to develop an effective means for rescue, treatment and prevention of ischemic diseases using the active ingredient tanshinone VI in Danshen.

[Means for Solving the Problems]

In order to achieve the above object, the gist of the present invention resides in a therapeutic agent for ischemic disease containing tanshinone VI as an active ingredient. Hereinafter, important matters related to the invention will be described. [Isolation of tanshinone VI] Tanshinone VI is commercially available dansin (Salvia miltiorrhiza)
Bge. Dried product) as a raw material, for example, by pre-extracting this with hexane to remove impurities, extracting with ethyl acetate, and then purifying using conventional purification means such as silica gel chromatography. (See FIG. 1). In addition to the desired tanshinone VI, tanshinone V having a similar structure was isolated and identified. §2 [Structure, physical and chemical properties of tanshinone V and VI] (A) Reagents and equipment used TLC-1 (chloroform), TLC-2 (hexane: ethyl acetate = 8: 2), TLC-3 (chloroform: Methanol = 1:
1; Sephadex LH20 (Pharmacia). Silica gel: Kieselgel 60 (70-230 mesh) by Merck; melting point: Yanagimoto melting point apparatus, ¹ HNMR JEOL PMX60; MS
Measurement: Shimadzu mass spectrometer GOKS-6020, IR measurement: Shimadzu IR
-420, UV measurement: Shimadzu spectrophotometer UV-260. (B) Chemical Structure As shown in the above formula, both are compounds in which quinone oxygen is bonded to the C ring of the phenanthrene ring, and the official chemical name of tanshinone VI is 1-hydroxy-2- (α-hydroxymethylethyl) ) -8-Methylphenanthrene-3,4-dione. (C) Physical and chemical properties Tanshinone V Property: orange-red powdery Optical rotation: ▲ [α] ³⁰ _D ▼ + 5.0 ゜ Mass spectrum: m / z: 314 (M ⁺ ) Nuclear magnetic resonance spectrum: ¹⁺ HNMR (CDCl ₃ ) δ: 1.25 (6H, ge
_{m, CH 3), 1.64 (} 3H, d, J = 6, CH-CH 3), 3.19 (2H.m, CH 2 O
H), 3.87 (1H, m , CH-CH 3), 7.60 (1H, d, J = 8, C 9 H), 7.9
0 (1H, d, J = 8, C 10 H) 13 C-NMRδ: 65.3 (CH 2 OH), 142.4
(-C-OH). Tanshinone VI Properties: orange-red needles Melting point: 206-208 ° C Optical rotation: ▲ [α] ³⁰ _D ▼ + 47.0 ゜ Mass spectrum: m / z: 296 (M ⁺ ) Nuclear magnetic resonance spectrum: ¹⁺ HNMR (CDCl ₃ ) δ: 1.50 (3H, d,
_{J = 6, CH-CH 3} ), 2.75 (3H, s, C 8 -CH 3), 3.20 (2H.m, CH 2
OH), 3.80 (1H, m , CH-CH 3, 7.02-7.20 (3H, m, C 6, 7,
_{9 H), 8.20 (1H,} d, J = 8, C 10 H), 9.20 (1H, d, J = 8, C 5 H).
^{13 C-NMRδ: 63.7 (CH} 2 OH), 146.9 (-C-OH). §3 [Tanshinone V and VI for hypoxic myocardial model
(1) Experimental method In the experiment, rats (Wistar strain, ♂, body weight 150-200)
g) Using the isolated heart, Langendorff
Perfusion was performed by the method, and the beating state was maintained. The perfusate was Krebs-Henseueit solution (1.20 mM
KH ₂ PO ₄ , 1.25 mM CaCl ₂ , 1.20 mM MgSO ₄ , 4.80 mM KCl, 120 mM
NaCl, 25 mM NaHCO ₃ ), 11 mM glucose,
% Oxygen and 5% carbon dioxide or 11 mM mannitol was used, and the mixture was saturated with 95% nitrogen and 5% carbon dioxide, respectively. The former solution was used for preperfusion and reoxygenation, and the latter solution was used for hypoxia load. The isolated heart was perfused at 7 ml / min and driven at a rate of 270 beats / min. Parameters such as cardiac contractility and perfusion pressure were recorded by a Nihon Kohden polygraph. Test drug administration method: Each drug (1.80 × 10 ⁴ M to 10 ³ M)
It diluted 10 to 10 ⁶ fold and administration of these dilutions into 30~100μ increments perfusate. The drug was dissolved in 10 to 50% of propylene glycol, which was further diluted with water. Infusion pump (Termo model STC521) for administration
Was administered into the perfusate at a rate of 0.1 ml / min. Hypoxia load and reoxygenation method: Hypoxia load to the isolated heart was performed by perfusion with the above mannitol-added, 95% nitrogen and 5% carbon dioxide saturated Krebs-Henseleit solution for 15 minutes. Cardiac function recovery by reoxygenation was performed for 45 minutes. Monitoring: During hypoxic load and reoxygenation, cardiac contractility, resting tension and coronary perfusion pressure were continuously recorded. In addition, release of creatine kinase enzyme and release of ATP metabolite from myocardial tissue were determined as indicators of increased permeability of cardiomyocyte membrane. For this reason, the effluent from the heart was collected over time, and the creatine kinase activity and the absorbance at 250 nm of the perfusate were measured using a spectrophotometer. (2) Pharmacological action of tanshinone V and VI (A) Direct action of drug on isolated perfused heart Tanninone V and tanshinone VI both have a slight suppression of cardiac contractility and an increase in perfusion pressure (both within 10%) at a concentration of 10 nmol. Admitted. At 100 nmol, a 20% reduction in perfusion pressure and strong suppression of cardiac contractility (35-50%) were observed. In general, some suppression of cardiac contraction may be preferable for the treatment of ischemic heart disease, but if the degree of suppression is too strong, it is not therapeutically preferable. Therefore, clinically, it appears that both may be used at concentrations of 10 nanomolar or less. (B) Pharmacological action on hypoxia-loaded heart When hypoxia is applied to the isolated perfused heart, the systolic force rapidly decreases immediately after the load is applied, and at the same time, the resting tension starts to increase. After disappearance, a sharp increase in resting tension and perfusion pressure was observed. Although reoxygenation tends to return the perfusion pressure to levels before hypoxia, cardiac contractility and resting tension hardly recover after 45 minutes of reoxygenation, and cardiac function remains arrested. In this pathological model, the systolic force after 45 minutes of reoxygenation was 21% of the initial value, the perfusion pressure was also 106%, and the resting tension was also 150.0%. Also, the amount of UV absorbers from myocardial tissue during hypoxia and reoxygenation increased significantly. This increase has been determined to be due to the release of ATP metabolites (S. Takeo et al., J. Pharm. Exp. Ther. 24
3,1131,1987). However, when tanshinone VI is added to the perfusate at the same time as hypoxia, significant recovery of myocardial contractility is observed during subsequent reperfusion. This is considered to indicate the cardioprotective effect of this drug during hypoxic load. That is, tanshinone VI has a 48-mole contractile force after reoxygenation by continuous infusion during a hypoxic load period of 300 pmol / min.
% Recovery (FIG. 2) and significantly suppressed the release of ATP metabolites into the perfusate as compared to no administration (FIG. 3).
Figure). However, no increase in static tension was observed (FIG. 4). Perfusion pressure was unaffected by administration of tanshinone VI during hypoxic loading, but during the reoxygenation period there was a clear tendency to promote the restoration of the original level (FIG. 5). In addition, the amount of acetin kinase released from the myocardium, which is an index of cell membrane permeability enhancement, was suppressed by 17% at 20 minutes of hypoxia and 12% at 45 minutes of reoxygenation. The content of high-energy phosphate compounds (ATP and creatine phosphate) 45 minutes after the start of reoxygenation was significantly recovered by administration of tanshinone VI (No. 6).
Figure). It has been reported by Takeo et al. That the magnitude of cardiomyocyte damage due to hypoxic load is estimated by the amount of ATP metabolite in the perfusate (S. Takeo and M, Sakanishi, J. Mol. Ce).
ll Cardiol. 15,577-594,1083 and Takeo et al. Can. JC
ardiol. 1988, printing). ). Probably, Tanshinone VI
By acting directly on the cell membrane during hypoxia load, it prevents the increase in membrane permeability and suppresses the release of the substrate required for ATP regeneration, thereby promoting the energy production of the myocardium during reoxygenation. Will play. (C) Conclusion From the above experimental facts, the drug of the present invention is expected to be effective as a drug having a prophylactic, therapeutic or rescue effect against ischemic conditions due to chronic stenosis or rapid spasm of coronary artery blood vessels. . As an administration method, injection into an artery or infusion into a vein are appropriate.

【Example】

According to the Japanese Pharmacopoeia General Rules for Preparations, Injection (A-107), 10 ml containing 2% tanshinone VI for injection and 20 ml
Each drug for infusion was prepared. The above-mentioned drugs are usually administered by injecting a single tube into a vein for ischemic disease in the case of injection, or one or two tubes in the infusion for infusion.

【The invention's effect】

As described above, the present invention has been used as a physiologic agent (blood circulation improving agent) in traditional Chinese medicine for the treatment of madness and myocardial infarction. The elucidation of the unknown active ingredient of ginseng and the possible clinical application of only the medicinal ingredient excluding its toxic ingredient can contribute to the maintenance or promotion of human health.

[Brief description of the drawings]

FIG. 1 is a process chart showing an example of the production process of tanshinone V and tanshinone VI, FIG. 2 is a graph showing the recovery of myocardial contractility by tanshinone VI, and FIG. Graph showing suppression of absorption amount, fourth
The figure shows the effect of tanshinone VI on cardiac resting tension, FIG. 5 shows the effect of tanshinone VI on the recovery of cardiac perfusion pressure, and FIG. 6 shows the effect of tanshinone on the high-energy phosphate compound content in the dried myocardium. 6 is a graph showing the influence of VI.

Claims (1)

(57) [Claims] [1] A therapeutic agent for ischemic disease, comprising tanshinone VI as an active ingredient.