JP3275183B2 - Infusion solution and method for producing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an infusion solution and a method for producing the same, and more particularly to an infusion solution containing neotrehalose as an active ingredient and a method for producing the same.

[0002]

2. Description of the Related Art Infusions containing glucose or fructose have been known for a long time and have been used as an energy source for living organisms.

[0003] However, infusions containing monosaccharides such as glucose and fructose are about 5 W / V% isotonic with blood, and are usually used at this concentration. There is a drawback that the patient must be administered a very large dose, causing long hours of pain for the patient.

In order to improve this, for example, Japanese Patent Publication No. Sho 4
In JP-A-9-48724, an infusion solution containing maltose is proposed.

[0005] Maltose is a disaccharide, and an infusion containing it is isotonic with blood at about 10 W / V% and can supply energy at twice the concentration of a monosaccharide. Can be halved.

[0006] However, maltose is a reducing disaccharide and has higher storage stability than reducing monosaccharides such as glucose and fructose, but is inherently unstable due to its reducing property. In particular, it is unstable in the coexistence of other nutrients such as amino acids and vitamins, which makes it difficult to realize a complex infusion using other nutrients in combination.

[0007] Therefore, it has been desired to realize an infusion solution containing the stable disaccharide maltitol, trehalose, etc., which does not show a reducing property.

[0008] However, maltitol is
As described in JP-A-7-13699, it is difficult to be metabolized and used in a living body, and is actually used as a diet sweetener.

On the other hand, trehalose is also disclosed in
As is clear from the description of JP-A-3-240758, "It is hardly digested and absorbed by the human body and has a low calorie." And "It is hardly susceptible to the enzymatic action of amylase and the like." It is understood as a carbohydrate that does not become an energy source in living organisms. In addition, it is difficult to obtain this carbohydrate in large quantities, and its detailed study has not been conducted.

[0010] Therefore, an infusion preparation which is isotonic with blood at a high concentration, has high storage stability, and contains a carbohydrate which is an energy source in a living body has not yet been established.

[0011]

SUMMARY OF THE INVENTION There is a strong demand for the establishment of an infusion solution containing a carbohydrate which is an energy source in a living body, isotonic with blood at the highest possible concentration, and has high storage stability, and a method for producing the same. I have.

[0012]

Means for Solving the Problems The present inventors have continued research on carbohydrates which can be an energy source for living organisms. In particular, for the use as an infusion agent, we have intensively studied a stable disaccharide, trehalose, and its related substances which do not show a reducing property. As a result, surprisingly, neotrehalose (O-α-D-glucopyranosyl β-D-glucopyranoside, also known as α, β-trehalose) can be converted to trehalose (also called α, α-trehalose) or isotrehalose (also called β, β- Unlike trehalose), it is found that it is easily metabolized and used as an energy source in living organisms, and is a stable saccharide that is isotonic with blood at high concentrations and does not show reducing properties. The present invention was found to be extremely convenient as a saccharide for use, and an infusion solution containing neotrehalose as an active ingredient and a method for producing the same were established, thereby completing the present invention.

In the infusion of the present invention containing neotrehalose as an active ingredient, since neotrehalose is a disaccharide, energy can be replenished at twice the concentration of a monosaccharide, and the restraint time of a patient is reduced by half. be able to.

In addition, since the neotrehalose does not show a reducing property and has high storage stability, the infusion solution of the present invention, together with neotrehalose as an active ingredient, contains amino acids, vitamins, other saccharides, lipids, minerals and the like. Other nutritional substances,
It is also easy to use a physiologically active substance such as an enzyme, a hormone, or a cytokine in combination to obtain a more comprehensive nutritional infusion or an infusion with a higher therapeutic effect.

An infusion solution containing neotrehalose as an active ingredient and a method for producing the same are described by the present invention.

The method for producing neotrehalose used in the present invention is not limited. For example, JP-A-63-216492
JP-A No. 307054 discloses a method of reacting cyclodextrin synthase with starch or partially degraded starch described in Japanese Patent Application Publication No. 307054, and adding β-galactosidase to lactone neotrehalose described in Japanese Patent Application No. 307054. It can be produced by a method such as acting.

For use in the infusion solution of the present invention, neotrehalose is preferably purified as much as possible, and usually has a neotrehalose content of 80 W / W% or more, preferably 90 W / W% or more.
A pyrogen-free material having a W / W% or more is preferable.

The infusion solution of the present invention may contain neotrehalose as an active ingredient, and even if it is a single agent which is an aqueous solution of neotrehalose alone, it may contain amino acids, vitamins, A complexing agent may be used in which one or more substances selected from other nutrient substances such as other carbohydrates, oils and fats, and minerals, and physiologically active substances such as antibiotics, enzymes, hormones, and cytokines are used in combination.

Neotrehalose is isotonic with blood at about 10 W / V%. The neotrehalose concentration of the infusion solution may be appropriately selected depending on various purposes, such as whether it is a single agent, a complex agent, or at the same time, is combined with another drug, an infusion agent, and the like. , From about 1 to 40 W / V%, more preferably from about 2 to 30 W / V%.

In the preparation of an infusion containing neotrehalose, crystalline neotrehalose is usually dissolved in water such as distilled water for injection, and then decolorized, desalted, and treated in a conventional manner.
Make it pyrogen-free by microfiltration, etc., mix other nutrients, physiologically active substances, etc. as necessary, and further, aseptically fill containers such as bottles and bags whose capacity is selected from, for example, about 10 to 500 ml. Sealing is performed, and if necessary, sterilization is performed.

The infusion thus obtained is administered parenterally, for example, intravenously, intraperitoneally, intramuscularly, by infusion or injection once or several times a day, without concern for toxicity and side effects. It is well-metabolized and can be advantageously used for replenishing energy to living organisms. The dose of neotrehalose is appropriately selected from the range of about 1 to 500 g, more preferably about 10 to 300 g, per adult day.

The infusion solution containing neotrehalose of the present invention, unlike glucose, requires almost no insulin for its metabolic utilization, and thus lacks insulin secretion as well as healthy subjects. Diabetics, the elderly with relatively low insulin levels,
For example, it can be advantageously used for energy supply to a diabetic or the like who is temporarily damaged due to a fracture, a cut, a burn, an operation, and has a shortage of insulin.

The infusion solution of the present invention can be used not only for humans but also for domestic animals such as cattle and horses, and pets such as dogs and cats, if necessary.

Hereinafter, the present invention will be described in detail by experiments.

(Experiment 1) In vitro digestibility test Using crystalline neotrehalose prepared by the method of Reference Example 1-2, Okada et al., Journal of the Japan Nutrition and Food Society, Vol. 43, No. 1, In accordance with the method reported on pages 23 to 29 (1990), neotrehalose was used as an aqueous solution, and an in vitro (in vitro) digestibility test was performed on the neotrehalose. As a decomposition rate (%).

[0026]

(Equation 1) The results are summarized in Table 1.

[0027]

[Table 1]

As evident from the results in Table 1, neotrehalose is well digested by small intestinal mucosal enzymes.

Next, digestion tests of various disaccharides were similarly performed using the small intestinal mucosal enzyme. The results are summarized in Table 2.

[0030]

[Table 2]

As is evident from the results in Table 2, it was surprisingly found that neotrehalose was digested by maltose in the intestinal mucosa enzyme second only to a greater extent than sucrose.

(Experiment 2) In Vivo Utilization Test (Experiment 2-1) Using crystalline neotrehalose prepared by the method of Reference Example 1-2, Atsuharu et al., Clinical Nutrition, Vol. 41, No. 2, 20th
According to the method reported on pages 0 to 208 (1972), 30 g of neotrehalose was used as a 20 W / V% aqueous solution, which was used by three volunteers (healthy, 26 years old, 39
Age, 52-year-old man) orally, blood is collected over time,
Blood glucose and insulin levels were measured. Glucose was used as a control.

As a result, neotrehalose showed the same behavior as that of glucose, and showed the same maximum value of both blood glucose level and insulin level about 0.5 to 1 hour after oral administration. Therefore, it was found that neotrehalose is easily digested, absorbed, metabolized, and used as an energy source.

(Experiment 2-2) Neotrehalose prepared by the method of Reference Example 1-2 was made pyrogen-free according to a conventional method, and this was replaced by Matsuzaki in Pharmaceutical Therapy, Vol. 6, No. 2, 65- Page 71 (1973)
Neotrehalose 5 according to the method reported in
0 g of a 10 W / V% aqueous solution was administered intravenously to two volunteers (healthy 37-year-old and 49-year-old men) by intravenous drip, blood was collected over time, and blood glucose level, insulin level, and neonatal level were measured. Urinary excretion of trehalose was measured. Glucose and maltose were used as controls.

As a result, while glucose showed a rapid increase in blood sugar and insulin levels, neotrehalose showed a similar tendency to that of maltose, that is, a slight increase in blood sugar levels, indicating that insulin levels were lower. There was little climb. The amount of urinary excretion is 10% of the dose
, Whereas neotrehalose and maltose were less than 20% of the dose. Therefore, neotrehalose is a sugar that is frequently metabolized and used, and is suitable for use as an infusion solution.

(Experiment 3) Acute toxicity test The crystalline neotrehalose prepared by the method of Reference Example 1-2 was orally administered to a 7-week-old dd mouse, and an acute toxicity test was performed. No deaths were observed up to 5 g per dose, and further administration was difficult. Therefore, the toxicity of this substance is extremely low.

Hereinafter, a production example of neotrehalose used in the present invention will be described with reference to a method described in Japanese Patent Application No. 307054 by the present inventors as a reference example.

(Reference Example 1) Preparation of Neotrehalose (Reference Example 1-1) Preparation of Lactonetrehalose 50 parts by weight of commercially available lactose and dextrin (DE8, sold by Matsutani Chemical Industry Co., Ltd., trade name: Paindex # 1)
Fifty parts by weight were dissolved by heating in 150 parts by weight of water, the solution was adjusted to a temperature of 60 ° C. and a pH of 6.0, and cyclomaltodextrin glucanotransferase derived from Bacillus stearothermophilus (Hayashibara Biochemical Laboratory Co., Ltd.) Was added and reacted for 20 hours and then heated to 100 ° C. for 30 minutes to inactivate the enzyme. The solution was adjusted to a temperature of 55 ° C. and a pH of 5.0, and glucoamylase (trade name: Glucoteam, sold by Nagaya Seikagaku Co., Ltd.) was added at a rate of 1 g / dextrum rum.
Add 5 units and allow to react for 16 hours.
The enzyme was inactivated by heating for minutes. This solution contains a novel oligosaccharide, lactone neotrehalose, at about 24 W /
W% was contained. This solution is decolorized with activated carbon, desalted and purified with an ion exchange resin (H type and OH type), concentrated to a concentration of about 45 W / W%, and subjected to column chromatography to contain a high content of lactone neotrehalose. Fractions were collected. The resin for fractionation is a salt type strongly acidic cation exchange resin (available from Organo Corporation, trade name: Amberlite XT-10)
16Na type), and packed in a water-suspended form into a jacketed stainless steel column having an inner diameter of 5.4 cm. At this time, four columns having a resin layer length of 5 m were continuously connected to each other to reduce the total resin layer length by about
m. While maintaining the temperature in the column at 55 ° C., the sugar solution of the raw material was added at 5 V / V%, and hot water at 55 ° C. was flowed at an SV of 0.3 to fractionate the fraction, and a lactone neotrehalose-rich fraction was collected. did. A part of the lactone neotrehalose-rich liquid (containing about 67 W / W% lactone neotrehalose per solid) collected by repeating this method was taken, concentrated to a concentration of 75 W / W%, and allowed to stand at 20 ° C. overnight. A crystal was deposited. The crystals were added as a seed crystal to a solution obtained by concentrating the lactone neotrehalose-rich solution to a concentration of 70 W / W%, and the resulting solution was subjected to auxiliary crystallization with slow stirring.
The resulting mass kit is disintegrated and the crystals are sprayed with a small amount of water and washed to obtain high purity crystals, which are dissolved in water, treated similarly and recrystallized to give a purity of 99.8 W / W
%, And about 3 parts by weight of very high purity crystals were collected.

Reference Example 1-2 Preparation of Neotrehalose 1 part by weight of the crystalline lactone neotrehalose obtained by the method of Reference Example 1-1 was dissolved by heating in 30 parts by weight of water.
H4.5, β-galactosidase (available from K-I Kasei Co., Ltd., trade name: Lactase LP) was added in an amount of 10 units per gram of lactone neotrehalose, and allowed to react for 20 hours. Deactivated. This solution contains neotrehalose and galactose at about 66 W / W% and 33 W / W per solid, respectively.
%. This solution was decolorized, desalted, purified and concentrated in the same manner as in Reference Example 1-1, and concentrated, and subjected to column chromatography using a salt type strongly acidic cation exchange resin to collect a neotrehalose-rich fraction.

A portion of the obtained neotrehalose-rich liquid (containing about 88 W / W% of neotrehalose per solid) was taken, concentrated to a concentration of 75 W / W%, and allowed to stand at 20 ° C. overnight. Deposited. The crystals were added as seed crystals to a solution obtained by concentrating the above-mentioned neotrehalose-rich solution to a concentration of 70 W / W%, and the crystals were auxiliary-crystallized with slow stirring. The resulting mass kit was separated, and a small amount of water was sprayed on the crystals. And washed to obtain high-purity crystals. The crystals are dissolved in water, treated in the same manner and recrystallized, and about 0.15 parts by weight of extremely high-purity crystals having a purity of 99.8 W / W% or more is obtained. Collected.

Hereinafter, embodiments of the present invention will be described.

[0042]

Example 1 Crystalline neotrehalose was added to water at a concentration of about 10 W
/ V%, then microfiltration according to a conventional method,
A plastic bottle was aseptically filled and plugged to obtain a product.

This product is a stable infusion without change over time,
It is suitable for intravenous or intraperitoneal administration for energy supplementation.

[0044]

Example 2 Crystalline neotrehalose was added to water at a concentration of 20 W /
V%, followed by microfiltration according to a conventional method, aseptically filling a plastic bag and plugging to obtain a product.

This product is a stable infusion without change over time,
For energy supplementation, it is convenient to mix with other drugs, infusions and the like at the same time and administer it at an appropriate concentration.

[0046]

Example 3 Crystal neotrehalose and crystalline sorbitol were mixed and dissolved in water at 5 W / V% and 2.5 W / V%, respectively, and then filled and sealed in a bottle as in Example 1. And got the product.

This product is a stable infusion without change over time,
It is suitable for intravenous or intraperitoneal administration for energy supplementation.

[0048]

Example 4 Crystalline neotrehalose and an amino acid compound having the following composition were mixed and dissolved in water at 5 W / V% and 3.0 W / V%, respectively, and then placed in a bag as in Example 2. The product was obtained by filling and plugging. Composition of amino acid formulation L-isoleucine 180 (milligram / 100
ml) L-leucine 410 L-lysine hydrochloride 620 L-methionine 240 L-phenylalanine 290 L-threonine 180 L-tryptophan 60 L-valine 200 L-arginine hydrochloride 270 L-histidine hydrochloride 130 glycine 340

This product is a stable infusion with no change over time because neotrehalose does not show a reducing property despite the complex infusion of carbohydrate and amino acid. It is suitable for intravenous and intraperitoneal administration for amino acid supplementation and energy supplementation before and after surgery.

[0050]

Example 5 Neotrehalose 10W / V% aqueous solution 10
A mixture obtained by adding 5 parts by weight of soybean oil and 1.5 parts by weight of egg yolk lecithin to 0 parts by weight was subjected to a mixer to obtain a coarse emulsion, which was then subjected to a pressurized injection emulsifier (manton-Gaulin Co., Ltd.) in a nitrogen stream at 600 kg / kg. The mixture was homogenized in cm ² to obtain an oil-in-water type fine particle emulsion having an average particle diameter of 0.2 μm or less, and then filled and plugged in the same manner as in Example 2 to obtain a product.

This product is a stable infusion that does not change over time, despite being a complex infusion of carbohydrates and lipids. It is suitable for intravenous, intraperitoneal, etc. administration for high energy supplementation. It is.

[0052]

Example 6 In a 10 W / V% aqueous solution of neotrehalose,
As a mineral, sodium dihydrogen phosphate 0.136W
/ V%, potassium acetate 0.098 W / V%, magnesium chloride hexahydrate 0.031 W / V%, calcium chloride
0.022 W / V% dihydrate and 1.5% sodium lactate
The resulting aqueous solution was adjusted to pH 5.5 by adding and dissolving it to 9 W / V%, and then filled and plugged in the same manner as in Example 2 to obtain a product.

This product is a stable infusion without change over time.
It is suitable to be administered intravenously, intraperitoneally, etc. for energy supplementation and mineral supplementation.

[0054]

Example 7 Neotrehalose 20W / V% aqueous solution
Human serum albumin 0.1 W / V% and human interferon-α were contained at 100,000 international units per ml, microfiltered, and then aseptically filled into 10 ml bottles and stoppered to obtain the product.

In this product, neotrehalose contributes to the improvement of the stability of the physiologically active substance interferon-α.
If stored in a cool, dark place, activity will be maintained for a long time. This product may be used in combination with distilled water for injection or other infusions for business purposes, and can be advantageously used to achieve the purpose of replenishing energy together with the therapeutic effect of the physiologically active substance.

[0056]

As is evident from the above, the infusion solution containing neotrehalose as an active ingredient of the present invention is:
Neotrehalose is isotonic with blood at a concentration of 10 W / V% and can supply energy at twice the concentration of monosaccharides, so the administration time can be halved and the patient's restraint time is greatly reduced. is there.

In addition, since the neotrehalose is stable without showing reducing properties, the infusion of the present invention can be used in combination with other nutrients and physiologically active substances together with neotrehalose to provide a more comprehensive nutritional infusion. Or an infusion with a higher therapeutic effect can be advantageously implemented.

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int.Cl. ⁷ , DB name) A61K 9/08 A61K 47/26

Claims (2)

(57) [Claims] 1. A method for producing an infusion solution containing neotrehalose as an active ingredient, comprising purifying crystalline neotrehalose as an aqueous solution, filling the solution into a container, and obtaining an infusion solution. 2. Nutrition along with neotrehalose
Contains at least one substance selected from the group consisting of
The active ingredient according to claim 1, wherein the active ingredient is provided.
To manufacture an infusion solution containing neotrehalose
Law .