JPS59110621A - Diuretic - Google Patents

Abstract

PURPOSE:To prepare a diuretic useful for the prevention and remedy, etc. of edema and ascites caused by nephropathy, by using 1-kestose, nystose and 1F fructofuranosyl nystose as active components. CONSTITUTION:The objective diuretic contains the compound of formula composed of sucrose (GF) consisting of glucose (G) bonded with fructose (F) and 1- 3 molecules of fructose bonded to the fructose fragment of said sucrose through beta2-1 bond, i.e. 1-kestose (n=2), nystose (n=3) and 1F fructofranosyl nystose (n= 4), as active components. The present fructo-oligosaccharide diuretic exhibits strong diuretic effect by a mechanism different from that of the conventional diuretic, and ameliorates the edema caused by various diseases such as cardiopathy, nephropathy such as acute hepatitis, renal insufficiency, nephrosis, etc. The compound is also expected to prevent the hepatic coma and uremia.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a diuretic, and more particularly to a diuretic whose main component is fructooligosaccharide, in which one to four molecules of fructose are bound to sucrose. More specifically, the present invention provides sucrose (GF) in which glucose (G) and fructose (F) are bonded, and 1 to 3 molecules of fructose are bonded to the 7-lactose moiety of the sucrose through β, -1 bonds. The compound 1-kesdose<GF2), which is specified by the following molecular weight and structural formula,
The present invention relates to a diuretic containing fructofuranosylnisdose (GF4) as an active ingredient.

Name Molecular formula Molecular weight Abbreviation Structural formula nl-
Kesdose C1811820□6504 GF2
2nisdose CQ4H4□0゜1666 0F93 The method for producing the above fructooligosaccharide is disclosed in JP-A-56-15496.
No. 7, No. 57-12973, and can be obtained by reacting Aureobasidium pullulansferase with sucrose. The sugar composition of fructooligosaccharide varies depending on the manufacturing conditions, but for example, the enzyme is added to a 60% solution of sucrose at pH 5, 0, and 4°C at 60°C.
When allowed to act for 8 hours, a reaction product having the following composition is obtained.

That is, fructose (F) 3%, glucose (G)
35%, sucrose (GF) 10%, 1-quesdose (GF2) 23%, nisdose (GFII) 23%
, IF-fructofuranosylnisdose (GF4) 6
%.

To purify fructooligosaccharides from this reaction product,
For example, 1 kg of the reaction product was packed into a 3 ol active column, and then mixed with 20 On' of water and then 1 kg of 3% ethanol solution.
801 was passed through and F, G, and GF were eluted.

Remove. Thereafter, the fructooligosaccharides (a mixture of GF2, GFQ and OF4) are recovered with 50 fi of 20% ethanol solution. This fructooligosaccharide is 1jQj M
Fractionated using a high performance liquid chromatograph, OF2゜0
F8 and GF4 are each isolated.

Next, GF and GF8 are dissolved in methanol to a concentration of 70% and left at room temperature for 7 days to precipitate and collect their respective crystals.

GF4 is isolated and eluted using a preparative high-performance liquid chromatograph, and the solution is freeze-dried to form a powder.

With the above steps, GF21109, GF81209
.

GF440 is obtained.

Next, the effects of these fructooligosaccharides will be explained.

Edema occurs due to various diseases such as axillary disorders and renal disorders such as acute nephritis, renal failure, and nephrosis.

Edema often worsens the underlying disease and is also known to cause secondary disorders.

Currently, ziazide drugs are used to treat edema.

Ethacrylic acid, furosemide, subiranolactone, and osmotic diuretics are known and are frequently used to treat edema.

The diuretic of the present invention containing fructooligosaccharide as an active ingredient is
It has a strong diuretic effect with a different mechanism of action than any of the diuretics mentioned above. For example, in an experiment using rats, which are experimental animals, a diuretic effect was expressed with a dose of 1 μg of 5C when administered intravenously, and with a dose of so O, the amount of urine and the amount of sodium and chloride up to 3 hours after administration were invention 7
The increase was approximately 10 times that of the group without lactooligoglycodiuretics,
The amount of sodium and chloride in the blood is also significantly reduced. Its diuretic strength is 16 times that of mannitol, which is clinically used as a glucodiuretic.

Furthermore, in various cases of hypertension, diuretics are often used to excrete sodium from the body to improve hypertension, and thiazides are preferably used as diuretics in this case. By the way, since the fructooligoglycodiuretic of the present invention also has a strong sodium excretion effect as described above, it can also be used as a diuretic for the purpose of treating hypertension.

During severe liver damage and renal failure, components and waste products that are not normally found in the body, such as ammonia, creatinine, and urea nitrogen, increase or accumulate in the body, leading to general fatigue, weakness, lethargy, and coma. be done. By applying the fructooligoglycodiuretic of the present invention to such symptoms, the above-mentioned blood waste products can be significantly reduced. For example, when administered intravenously to rats, which are experimental animals, the effect appears from 501μ%, and so
The amount of yAg increased the amount of urea nitrogen excreted into the urine by about 5 times compared to rats not treated with fructooligoglycodiuretics.
It also increases the urinary excretion of creatinine by about 7 times.

It is known that hypercalcemia develops due to diseases such as primary, sexual hypoparathyroidism, pseudoparathyroidism, and hyperthyroidism. As the disease progresses, hypercalcemia causes general malaise, polydipsia, polyuria, constipation, poor eating habits, vomiting, drowsiness, and weakness.

Symptoms worsen in the order of confusion, first, and then coma, so treatment is necessary. Currently, EDT is used as a treatment for high calcium.
A, sodium sulfate, mitalamycin, furosemide, steroids, calcitonin, etc. are known.

The fructooligoglycodiuretic of the present invention has a mechanism of action different from that of any of the above-mentioned drugs, and is capable of strongly excreting calcium into the urine and lowering blood calcium levels. For example, in experiments using rats, intravenous administration of 50
Hiroshi ~ The effect is expressed by the amount 1. ．． When administered at a dose of 500 m9A, i9, the average blood calcium level of rats without drug administration was 9.
6 my/n to 8.7 mQAg average.

In addition, in the clinical use of fructooligosaccharide, it is possible to excrete it into the urine by a route other than the oral route, preferably intravenously. It must be administered within an unlimited amount of time.Since the LD5o value of GF2, GFa, and GF4 after a single intravenous administration in rats at 100°C is 4 or more, it can be administered without limit. It is.

In addition, the above-mentioned GF2 has the strongest effect among fructooligosaccharides, so it is preferable to use GF2 for clinical use.

For practical use, it is desirable to administer 20 to 30 doses once or twice a day.
00M or should be done slowly as a concentrated solution. In addition, blood and cormorant ``11-''?i pressure is approximately 2
It is 0%.

Fructooligosaccharide, especially GF, has a diuretic effect (because it is strong, it causes the excretion of not only sodium, chloride, and waste materials, but also calcium and, although weakly, water, so it is used for the treatment and prevention of edema and the promotion of waste excretion from the body. If the diuretic of the present invention is used in a significantly large amount, calcium and potassium supplementation may be required.Also, the diuretic of the present invention may be administered in a significantly large amount for the purpose of treating and preventing hypercalcemia. In some cases, supplementation of sodium and potassium may be necessary for the same purpose as above.

As described above, the fructooligoglycodiuretic of the present invention is a diuretic that exhibits a strong diuretic effect through a mechanism of action that is different from that of conventional diuretics. In the future, it is expected to be effective in the treatment and prevention of patients with edema due to various diseases, as well as in the prevention and treatment of hepatic coma and uremia, and is also expected to be effective in the treatment of hypertension in hospitalized patients. It is an extremely useful substance for maintaining human health.

The present invention will be explained below with reference to Examples.

Example 1 Wistar rats (SPF
) were used in groups of 3 to 6 males.

The animals were orally administered saline warmed to 37°C for 3 d, immediately followed by GF, 2. GF, , GF
a and 500 μm and 100 μm of mannitol as a control.
A single dose of 0 IV/ky was administered intravenously. All test substances were dissolved in distilled water for injection, and the concentration was adjusted so that the volume administered per animal was 1 ml.

Immediately after the completion of intravenous administration, the animals were placed in individual urine collection cages to collect and collect urine from 0 to 3 hours and 3 to 6 hours. still,
During the urine collection period, the subjects were fasted and provided with water.

After the urine collection was completed, the animals were subjected to blood collection by cutting the cervical artery and vein. Serum was immediately separated from the collected blood.

The following items were measured for serum and urine.

Urine: Urine volume, sodium, potassium, chloride, calcium, urea nitrogen, osmolarity, and creatinine were measured.

Serum: The same items as above except for urine volume and osmotic pressure were measured.

The results are shown in Tables 1 and 2 below.

Example 2 Wistar rats (SPF
) were used in groups of 8 males.

The animals were orally administered 3N physiological saline solution warmed to 37°C. Immediately after that, GF2's 50.100°200,
400. 800 m9/Ig and, as a control, 9t of mannitol was administered once intravenously at 800 m9/Ig. In each case, the concentration of the test substance was adjusted with distilled water for pharmacopoeial injection so that the amount was 1 d/rat.

Immediately after administration of the test substance, the animals were placed in individual urine collection cages, and urine was collected for 0-3 hours and 3-6 hours.

In addition, fasting water was provided during the urine collection period.

After the completion of urine collection for 6 hours, the cervical artery and vein of the test animal was cut and blood was collected. Blood was immediately spun down and serum was collected.

For urine, osmotic pressure, urea nitrogen, creatinine, urinary volume, sodium, potassium, and chloride were measured, and for serum, sodium, potassium, and chloride were measured.

The results are shown in Table-3 and Table-4.

Control 8 100 100 100 100100GF
250 8 101 92 100 98
1000F, 2100 8 99 105 10
0 97 1000F2200 8 101 9
7 100 97 990F2400 8 99
97 100 95 990F2800 8
95 119 99* 93 96** (Note) Significant difference compared to control *...P<0.0.5
**...P<0.01 Example 3 Test animals were Wistar rats with an average weight of 224 mm) (,
5PF) males were used in groups of 2-3.

GF was dissolved in distilled water for pharmacopoeial injection at a concentration of 44.8%. The dose was 2gAg and 49/ig, and the drug was intravenously administered once into the tail vein. As a control, a group administered with 2 ml of distilled water was provided.

General conditions were observed up to 6 hours after administration and before and after body weight measurements. Body weight measurements were carried out at 1.3 and 9 days after administration.

The results are shown below.

(a) - Membrane condition: Immediately after administration, the animal became depressed, and 1 hour after administration, piloerection was observed, and from 15 to 2 hours after administration, a marked diuretic state (a large amount of urine was stored under the cage) was observed. The same thing happened at the time of kenken 6 hours later. 29/k
In group g, these symptoms recovered the next day, but 49/
In the kg group, it took 2 days to return to normal. Thereafter, the trend was similar to that of the control group.

(b) Weight change and fatal cases Before administration Day 1 Day 3 Day 9 Dead cases control group
100 106 116 133 0/2OF229
7kg 100 96 105 121
0/3GFa 4 gAg 100 90
95 111 0/3 Patent applicant Meiji Seika Co., Ltd. Procedural amendment (voluntary) October 19, 1980 Commissioner of the Patent Office Kazuo Wakasugi Tono 1, Indication of case Patent application 1982-220167 Z Name of the invention Diuretic 5 Amendment Relationship with the case of the person making the amendment Patent applicant Meiji Seika Co., Ltd. 4, agent 〒104 5 Column 6 for detailed explanation of the invention in the specification subject to amendment, Contents of the amendment (1) Page 8, line 9 of the specification [Hypercalcemia treatment agent] for eyes has been corrected to [Hypercalcemia treatment agent]! Ru.

(2) "A, 7 mg/Yu" in the fifth line from the bottom of page 8 is corrected to 18,7 mg/di J.

(3) Replace “ay, 1o o j” with “GF410o
o” and correct it.

(4) Delete all columns of uric acid (0 to 5 hours) in Table 2 on page 15.

(that's all)

Claims (1)

[Claims] 1. Sucrose (GF) in which glucose (G) and 7-lactose (F) are combined, and 7-lactose of the sucrose
Fructose is attached to the lactose moiety by β2-1 bond.
1-kesdose < GF
2), Nisdose (GFa). A diuretic containing 1F fructofuranosylnisdose (CGF4) as an active ingredient. Name Molecular formula Molecular weight abbreviation Structural formula ○1-
Kesdose Cl8H8Q0□65040F22 Varnishdose 024H4□0. □ 666 GF, 2. For the prevention and treatment of edema and ascites accumulation caused by various pathological conditions such as liver, kidney, and heart diseases, cancer, and for lowering cerebral pressure during head trauma. The diuretic according to claim 1, which is used for lowering intraocular pressure in glaucoma. 3. Accumulation of waste products in the body and liver damage due to kidney damage, etc.
The diuretic according to claim 1, which is used for the prevention and treatment of all disorders caused by an increase in blood of components that are constant in the body or components that are not normally found in the body due to diseases such as cancer.