JPS626688B2 - - Google Patents

Description

[Detailed description of the invention]

TECHNICAL FIELD The present invention relates to a vascular wall metabolism improving agent whose main component is unsaponifiable matter of soybean oil. Today, among cardiovascular drugs, various antilipidemic drugs have been developed for the purpose of treating hyperlipidemia. Examples of drugs that suppress
Only a few examples are known, such as Medical Tribune, Vol. 10, No. 45, 1977). Among these, elastase is an elastin-degrading enzyme that exists in the pancreas of mammals or the cells of specific bacteria, and there are difficult problems in obtaining raw materials, extraction, purification, stabilization, etc. accompanied by. Furthermore, there are still questions regarding whether long-term administration of Cetaben, whose chemical name is sodium p-dexadecylamino-e-benzoate, is possible due to side effects. As a result of intensive research in order to solve the above-mentioned problems, the present inventors have developed an unsaponifiable product of soybean oil derived from a natural product that is extremely cheap, easy to obtain raw materials, easy to manufacture, and has no particular side effects. Improves metabolism of arterial walls with effects on physical properties, i.e., improves metabolism of elastin, collagen, acidic mucopolysaccharides, glycoproteins, smooth muscle cell fibers, and smooth muscle cell nuclear DNA associated with sclerotic lesions of arterial tissues. In addition to having an improving effect,
It was discovered that it also has the effect of suppressing calcium deposition, and as a result, aging of blood vessels and arteriosclerosis can be effectively prevented, and based on this knowledge, the present invention was completed. The unsaponifiable soybean oil according to the present invention is a semi-solid or viscous oily substance that is obtained as a pot residue after molecular distillation by esterifying a deodorized distillate of soybean oil and has a unique odor and a slight sweetness. It exhibits a serum lipid effect (Takashi Kaneda et al., Nutrition and Food, 19, 439 (1967)), and usually contains 40 to 50% by weight of plant sterols (β-sitosterol, stigmasterol, campesterol) (hereinafter referred to as weight%). %), 18-22% tocopherols, and about 30% higher unsaturated fatty acids (lauric acid, myristic acid, palmitic acid, linoleic acid, etc.). One example of purification of the above-mentioned soybean oil unsaponifiables is as follows:
Soybean oil is heated to approximately 230℃ using a guard type deodorizing device.
A deodorized distillate of soybean oil is obtained by blowing 5% steam for 1 hour under a reduced pressure of 2 to 3 mmHg. 1740g of this deodorized distillate, 3000g of methanol and concentrated sulfuric acid.
After adding 34 g of the mixture and refluxing at 68°C for 3 to 4 hours, excess methanol was distilled off under reduced pressure. Pour the residue into 5 kg of hot water.
After washing with water, completely distill off the moisture, and then
When fatty acid methyl is distilled off at 170-190℃ under a reduced pressure of μHg, plant sterols (β-sitosterol, stigmasterol, campesterol) 42
~43%, tocopherols (α-, γ, δ-tocopherol) 18-22%, and higher unsaturated fatty acids (lauric acid, myristic acid, palmitic acid, linoleic acid, etc.) approximately 30%. The unsaponifiable matter of soybean oil is obtained as a pot residue. The specific conditions for the above molecular distillation and the specific results corresponding to the conditions,
The results are shown in Table 1 below, and the infrared absorption spectrum of the purified product No. 1 in Table 1 is shown in FIG.

[Table] Each component of the above purified product was quantified by the following method. (A) Plant sterols It was determined from the weight of dichitonide produced by saponifying a sample with sodium hydroxide and reacting with dichitonin, and the relative ratio of campesterol, stigmasterol, and β-sitosterol obtained from a gas chromatogram. (B) Tocopherols Dissolve the sample in chloroform, add a solution of ferric chloride in anhydrous ethanol, and then add α, α′-dipyridyl to form a red complex salt.
The absorbance at 520 mμ was measured, and the absorbance was determined from each absorbance using α-tocopherol as a standard solution. (C) Higher unsaturated fatty acids Calculated from the formula (saponification value/neutralization value) x 100 (%). However, since the fatty acids in the sample do not always have a constant composition, the neutralization value of linoleic acid was used as a substitute for convenience. According to the experiments conducted by the present inventors, the above-mentioned unsaponifiable soybean oil has a particularly excellent effect of improving blood vessel wall metabolism, and is also capable of extremely effectively preventing calcium deposition on the blood vessel wall. it is obvious. Originally, blood vessels under living body pulsating flow are constantly undergoing physiological, pathological, and compensatory changes, that is, the quantity and
Qualitative transformation is forced. In this case, elastin, collagen, smooth muscle fibers, and smooth muscle cell nuclei, which are components of vascular wall construction.
If it improves the metabolism of DNA, acidic mucopolysaccharides, and glycoproteins associated with sclerotic lesions, and prevents calcium deposition on the vascular wall, which is closely related to the physical properties of blood vessels, especially the viscoelastic properties of blood vessels, it will become a component of vascular wall construction. This is the reason why it suppresses the deformation of blood vessels and prevents aging of blood vessels and arteriosclerosis. From this point of view, the present inventors used Japanese white adult male rabbits, 36 months old and weighing 2.4 to 4.5 kg, to create a healthy group of 3 rabbits each, as shown below.
Experiments related to the present invention were conducted by forming a curing group and a curing dosage group. (A) Formation of cured group and cured drug group The following methods (1), (2), and (3) were used in combination to create cured rabbits. (1) Tissue damage method 0.1% adrenaline 0.13ml/Kg intramuscularly,
Administer once a day, 5 days a week, for 16 consecutive weeks. (2) Hypoxic method Nitrogen gas is spewed out from the inlet of a sealed box,
Air is removed from the outlet and the rabbit undergoes clonic convulsions due to lack of oxygen. Twice a day, 5 times a week
day, for 4 weeks. (3) Intima production method (vegetative vascular occlusion method) Cholesterol is administered orally at 0.5 g per day for 16 consecutive weeks. Next, in parallel with the production of cured rabbits, the cured administration group received 200 mg of unsaponifiable soybean oil containing 48% plant sterols, 18% tocopherols, and 30% higher unsaturated fatty acids orally. The drug was administered once a day for 22 consecutive weeks. (B) Preparation of Specimens and Measurement and Observation of Components of Blood Vessel Walls The healthy group, cured group, and cured drug group were sacrificed after 22 weeks, and each thoracic aorta was sliced into 3-5 mm wide slices, and then wrapped in epoxy resin. The specimen was embedded and sliced laterally using a JB-4A automicrotome to prepare specimens. Staining method of each component and histochemical microspectroscopy (Shozo Abe, Journal of Tokyo Jikei University School of Medicine, 90 , 1)
(1975)) are shown in Table 2 below.
Shown below.

[Table] Notes (1) Values for each group (a), (b), and (c) are the average value of three birds ± standard error% Extinction, however, the unit of (d) is
AU. Values in parentheses are indexes based on the healthy group. (2) ※...p<0.2, ※※...p<0.05, ※※※...p<0.001 (all t-test) Blood vessel wall building components in the sclerosing medication group shown in Table 2 above The comparison between the values for and those of other groups clearly shows the vascular metabolism improving effect of the vascular wall metabolism improving agent of the present invention. Next, using each of the specimens, calcium deposits and elastic fibers, which are most closely related to the physical properties of arteriosclerotic blood vessels, were observed using an optical microscope, and the results are shown in FIGS. 2 and 3. FIG. 2 is an optical micrograph of a Kossa-Nishiyama stained specimen for calcium, in which (A) shows a healthy group, (B) shows a cured group, and (C) shows a cured group. FIG. 3 is an optical micrograph of a Weigert-stained specimen of elastic fibers, in which (A) shows a healthy group, (B) shows a hardened group, and (C) shows a hardened group. As is clear from Figure 3, in the hardened group, calcium deposition, disorder of elastic fibers, thinning, rupture,
In contrast, in the sclerosing treatment group, there was almost no difference from the healthy group.
It is extremely clear that the agent of the present invention has a special effect on suppressing calcium deposition and maintaining the viscoelastic properties of blood vessels. In short, the above experimental results show that the agent of the present invention has an unprecedented effect as a vascular wall metabolism improving agent, and together with its calcium deposition prevention effect, it is effective in preventing vascular aging and arteriosclerosis. It is shown that it has a significant effect. Moreover, since the agent of the present invention is mainly composed of unsaponifiable matter of soybean oil, it can be provided at an extremely low cost, and in this respect, it has great industrial applicability. The agent of the present invention is used as an oral preparation such as an emulsion, a powder, a tablet, a granule, or a capsule.
Since unsaponifiable soybean oil is a semi-solid or viscous oily substance, it is convenient to use it in solid form. In other words, hard encapsulation or soft encapsulation is appropriate. Next, examples of the present invention will be shown. Example 1 As a method for non-invasively measuring arteriosclerosis, the numerical value of aortic wave velocity (hereinafter abbreviated as PWV) well reflects the vascular physical properties derived from the construction status of the vascular wall material, and the PWV value The finding that the amount increases with the progression of sclerotic lesions (Shozo Yoshimura et al., Angiology, 18,
742 (1978)) by the PWV method (Genji Hasegawa, Tokyo Jikei University School of Medicine Journal, 85 , 742 (1970)).
42% plant sterols, 22 tocopherols
A clinical experiment was conducted using encapsulated unsaponifiable soybean oil containing 31% of higher unsaturated fatty acids. The hardened group was aged 45 to 81 years (average 65.3 years) and male.
A total of 35 patients, 19 cases and 16 women, were in the hardening treatment group, aged 33 and up.
A total of 34 patients, 79 years old (average 61.3 years), 17 males and 17 females, were orally administered the above-mentioned soybean oil unsaponifiables at a dose of 400 mg three times a day for 28 consecutive months. PWV was measured once every 4.4 months for both the hardening group and the hardening medication group, and the changes in the average values were examined, and the results are shown in Figure 4. As is clear from Fig. 4, there was no difference between the hardening group and the hardening treatment group for less than 10 months, P < 0.055 for 10 to 20 months, and P < 0.01 for 20 to 30 months. A significant difference was observed, and it was revealed that unsaponifiable soybean oil exhibits the effect of suppressing or inhibiting human arteriosclerotic lesions when administered for a period of more than 10 months. In addition, Figure 4 shows the average at month 0 for each group.
It is displayed based on the PWV value. Example 2 45% plant sterols, 20% tocopherols
%, unsaponifiables of soybean oil containing 30% of higher unsaturated fatty acids are dissolved in sesame oil, and the weight is 12~12% at 5 weeks of age.
18g DD mouse (female, male) and weight 60-90g
An acute toxicity test was conducted using 5 Wistar rats (female and male) in groups of 5 each. In mice, the soybean oil unsaponifiables were administered orally.
8.0g/Kg, subcutaneous injection 4.0g/Kg, intraperitoneal injection
For rats, the same unsaponifiable soybean oil as for mice was administered orally at 8.0 g/Kg, subcutaneously at 2.0 g/Kg, and intraperitoneally at 1.0 g/Kg. Life or death was determined 72 hours after administration, and observation was continued for 10 days. As a result, as determined 72 hours after administration, all of the animals survived, and there were no deaths during the subsequent 10 days, making it impossible to calculate _LD50 . Furthermore, no differences were observed in the toxic symptoms and behavior after administration compared to the normal animal group. Reference example: Soybean oil unsaponifiables 470g, vitamin C 20g, citric acid 10g, cellulose calcium glycolate 40g
g, sodium laurate 20g, polyoxyethylene monostearate 10g and chloroform 600g
ml and stir well to form a suspension. 390 g of Aerosil 200-400 (trade name) is added thereto, mixed and stirred, and then dried at a temperature of about 50-60 ℃, and the obtained lumps are crushed to obtain an adsorbed powder of soybean oil unsaponifiables. Add 600 ml of chlorocene-ethanol solution containing 40 g of polyvinylpyrrolidone to this adsorbed powder, and after kneading,
Granules were granulated and dried according to a conventional method to obtain granules with good solidification. A small amount of magnesium stearate is added to this product as a lubricant, and each capsule contains saponified soybean oil.
Fill into hard capsules to contain 200mg.

[Brief explanation of the drawing]

Figure 1 is an infrared absorption spectrum diagram of unsaponifiable soybean oil according to the present invention, and Figure 2 is an infrared absorption spectrum diagram of unsaponifiable soybean oil according to the present invention.
Optical micrographs of Kossa-Nishiyama stained specimens, in which (A) shows the healthy group, (B) shows the cured group, and (C) shows the cured group. Figure 3 shows elastic fibers.
These are optical micrographs of Weigert-stained specimens, in which (A) shows a healthy group, (B) shows a cured group, and (C) shows a cured group. FIG. 4 is a diagram of changes in aortic pulse wave velocity obtained by the PWV method.

Claims (1)

[Scope of Claims] 1. A vascular wall metabolism improving agent whose main component is unsaponifiable matter of soybean oil. 2. Soybean oil unsaponifiables are made into capsules,
The vascular wall metabolism improving agent according to claim 1.