KR102129366B1 - An infusion solution for treating inflammation - Google Patents

Abstract

Disclosed is a fluid composition for treating inflammation containing a specific sugar.

Description

Infusion solution for treating inflammation

The present invention relates to a fluid composition for the treatment of inflammation.

The main materials covered in the life sciences are proteins, amino acids, nucleic acids, fats, fatty acids, carbohydrates, polysaccharides and sugars. Life science and medical researchers have studied and researched protein as a major communication molecule in vivo through a long period of research, but came to the conclusion that protein alone is not sufficient to deliver all messages inside cells, and accordingly, protein molecule and carbohydrate molecule Began to study the bound glycoproteins.

Glucose (Glu), Galactose (Gal), Mannose (Man), L-Fucose (Fuc), Xylose (Xy), N-acetylglucosamine (N-Acetylglucosamine: GlcNAc), N-Acetylgalactosamine (N-Acetylgalactosamine: GalNAc), N-Acetylneuraminic acid (NANA; or Sialic acid) Plays a very important role in

The present inventors confirmed that the infusion composition containing 8 kinds of sugar as described above has an effect of treating inflammation, and completed the present invention.

Korean Patent Publication No. 10-2011-0131699

The present invention is to provide an infusion composition for the treatment of inflammation comprising a specific sugar.

In order to solve the above problems, the present invention discloses a solution composition for treating inflammation containing a specific sugar.

Specifically, the infusion composition of the present invention includes glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine, and sugars of N-acetylneuraminic acid.

The infusion composition of the present invention may further include one or more amino acids.

Here, the amino acid is at least one selected from the group consisting of isoleucine, leucine, lysine acetate, methionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, proline, serine, tyrosine, and glycine. Can be.

Alternatively, as the amino acid, one or more of 20 amino acids capable of constituting a human protein may be selected.

The infusion composition according to the present invention exhibits an excellent inflammatory treatment effect.

1 is a graph showing the swelling rate of the foot 3 hours after the administration of the inflammatory agent in the edema inhibition experiment.
2 to 4 are graphs showing the results of experiments in suppressing the effects of psychokines.

The present invention will be described in detail based on the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1: Preparation of sugar-containing solution

Glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine, and N-acetylneuraminic acid from Sigma-Aldrich (St. Louis, Missouri, USA) 8 sugars were dissolved in water for injection (foreign pharmaceutical). As a stabilizer, aspartic acid (L-Aspartic acid), which is a type of amino acid, was used.

In order to adjust the pH of the prepared sugar-containing aqueous solution, a 1N NaOH solution (Samjeon pure chemical Co., Ltd.) was used. The pH was adjusted to 7.0 using a pH meter (METTLER TOLEDO Seven Compact pH/Ion).

Table 1 shows the composition of the sugar-containing solution.

ingredient Sugar content Infusion Sugar (mg) L-(-)-Fucose 31 D-(+)-Galactose 31 D-(+)-Xylose 31 D-(+)-Glucose 31 D-(+)-Mannose 31 N-Acetylneuraminic acid 31 N-Acetyl-D-galactosamine 31 N-Acetyl-D-glucosamine 31 Stabilizer (mg) Aspartic acid 20 solvent Water for injection q.s. pH adjuster (μL) 1N NaOH q.s. Total 10 mL

* Total 10mL standard composition table (actually 50mL was prepared)

Example 2: Preparation of sap solutions containing sugars and amino acids

Glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine, and N-acetylneuraminic acid from Sigma-Aldrich (St. Louis, Missouri, USA) After dissolving 8 kinds of sugars in water for injection (foreign pharmaceutical), it was mixed with commercially available amino acid infusion solution, 89.4% furoamine (Proamin injection, Hanol Biopharma). The furoamine strain contains amino acids such as isoleucine, leucine, lysine acetate, methionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, proline, serine, tyrosine, and glycine. Aminoguanidine (Sigma-Aldrich) was used as a stabilizer.

To adjust the pH of the prepared aqueous solution containing sugar and amino acids, a 1N NaOH solution (Samjeon Pure Chemical Co., Ltd.) was used. The pH was adjusted to 7.0 using a pH meter (METTLER TOLEDO Seven Compact pH/Ion).

Table 2 shows the composition of the aqueous solution containing sugars and amino acids.

ingredient Sugars and amino acids
contain Infusion Sugar (mg) L-(-)-Fucose 31 D-(+)-Galactose 31 D-(+)-Xylose 31 D-(+)-Glucose 31 D-(+)-Mannose 31 N-Acetylneuraminic acid 31 N-Acetyl-D-galactosamine 31 N-Acetyl-D-glucosamine 31 Amino acid (mg) L-Isoleucine 28 L-Leucine 62.5 L-Lysine Acetate 62 L-Methionine 17.5 L-Phenylalanine 46.75 L-Threonine 32.5 L-Tryptophan 6.5 L-Valine 22.5 L-Alanine 31.25 L-Arginine 39.5 L-Aspartic Acid 19 L-Cysteine 5 L-Glutamic Acid 32.5 L-Histidine 30 L-Proline 16.5 L-Serine 11 L-Tyrosine 1.75 Aminoacetic Acid 53.5 Stabilizer (mg) Aminoguanidine 40 solvent Water for injection q.s. pH adjuster (μL) 1N NaOH q.s. Total 10 mL

* Total 10mL standard composition table (actually 50mL was prepared)

Example 3: Anti-inflammatory Effect test

end. Experimental method

Male rats (6-week-old SD rats, each weighing about 200g) are purchased so that solid feed and water can be freely consumed. 12 hours are brightened and 12 hours are darkened to manage light and constant temperature (22± 2°C).

Prior to the experiment, mice were divided into 8 groups of 3 animals per each experimental group and anesthetized with ether.

Before the experiment, the rat's foot thickness was measured with a Dial Caliper (Mitutyo, Japan).

In the control group, 100 μL of Normal Saline was administered subcutaneously to the sole of the foot.

As a positive control, a mixed solution with PBS solution was prepared so that the anti-inflammatory drug indomethacin was administered at a dose of 5 mg/kg, and 100 μL was administered subcutaneously to the bottom of the foot.

Six sample solutions, i.e., sugar and amino acid-containing solution stocks, sugar and amino acid-containing solution stocks 10-fold dilutions, sugar and amino acid-containing solution stocks, 100-fold dilutions, sugar-containing solution stocks, sugar-containing solution stocks, 10-fold dilutions, sugar-containing solution stocks Each 100-fold dilution was administered by subcutaneous injection to the soles of rats at 100 μL each.

The sample materials were pretreated, and after 1 hour, 100 μL of a 1% solution of carrageenan (a solvent: normal saline), an inflammation-inducing substance, was injected subcutaneously into the soles of the feet.

After carrageenan administration, the rat's foot thickness was measured. The volume of the rat foot was 1 hour before carrageenan administration, immediately after carrageenan administration, 1 hour after carrageenan administration, 2 hours, 3 hours, and 5 hours, respectively, using dial caliper (Mitutyo, Japan), respectively. The severe edema was measured.

I. Experiment result

As a result of rat foot edema evaluation, negative control group, positive control group (indomethacin), 6 sample substances (i.e., sap solution containing sugar and amino acid, 10x dilution, 100x dilution, sugar containing sac, 10x dilution) , 100-fold dilutions) Table 3 shows the results of the measurement of the foot thickness of the rats showing the effect of suppressing edema.

Rat foot bottom thickness (average value, in mm) R L R L R L R L R L R L 1 hour before carrageenan administration Immediately after carrageenan administration 1 hour after carrageenan administration 2 hours after carrageenan administration 3 hours after carrageenan administration 5 hours after carrageenan administration Negative control
(Control group) 0.514 0.529 0.608 0.613 0.683 0.676 0.711 0.645 0.727 0.724 0.686 0.678 Positive control
(Indomethacin group) 0.585 0.550 0.599 0.615 0.617 0.624 0.641 0.634 0.624 0.679 0.627 0.694 Infusion solution containing sugar and amino acid 0.595 0.581 0.702 0.616 0.621 0.612 0.633 0.676 0.611 0.651 0.611 0.618 10-fold diluent of sugar and amino acid-containing solutions 0.622 0.623 0.631 0.62 0.626 0.635 0.659 0.640 0.681 0.663 0.637 0.663 100-fold dilution of sugar and amino acid-containing solutions 0.578 0.593 0.699 0.682 0.651 0.638 0.669 0.649 0.664 0.687 0.636 0.622 Sugar-containing sap
Undiluted 0.524 0.542 0.712 0.635 0.612 0.722 0.684 0.654 0.659 0.657 0.677 0.651 Diluted 10 times the sugar-containing solution 0.610 0.585 0.685 0.634 0.672 0.697 0.648 0.671 0.663 0.694 0.672 0.662 100-fold dilution of sugar-containing sap solution 0.595 0.581 0.702 0.616 0.621 0.612 0.633 0.676 0.611 0.651 0.611 0.618

* SD Rat 6 weeks Male 3 experiments per sample, total 24 animals * R: Rat right foot L: Rat left foot

Foot edema rate (%) value after 3 hours after administration of the inflammatory substance carrageenan is shown in Table 4 and Figure 1 below:

sample Control group Indomethacin group number 3
Sugar/amino acid solution Number 4
Sugar/amino acid solution 10 times dilution Number 5
Sugar/amino acid solution 100 times dilution No. 6
Sugar sap
Undiluted Number seven
Sugar sap
10-fold dilution Eight
Sugar solution 100 times dilution foot
Edema rate (%) 6.5% 20.1% 2.7% 9.5% 14.9% 6.3% 12.3% 21.3%

As a result of the experiment, the effect of inhibiting the edema of the sugar and amino acid sap solution was much better than that of indomethacin, and the 10-fold and 100-fold dilutions of the sugar and amino acid sap also had better inhibitory effects than indomethacin.

The edema-inhibiting effect of the sugar-containing sap solution was much better than indomethacin, and the 10-fold dilution and 100-fold dilution of the sugar-containing sap also had better inhibitory effect than indomethacin.

In addition, the sap and sugar-containing sap and the sac-containing sap showed inflammation inhibitory effect in proportion to the concentration.

Therefore, it can be confirmed that the sap composition according to the present invention has an excellent anti-inflammatory effect.

Example 4: IL- 1βIL -6, PGE2 Expression inhibition effect experiment

end. Experimental method

Inhibition of IL-1βIL-6 and PGE2 expression suppression experiments on the sugar and amino acid sap extracts and sugar-containing sap extracts with the lowest paw edema rates, and the negative control sample (normal saline) and the positive control sample (indomethacin). It was carried out.

Measurement of IL-1βIL-6 and PGE2 was performed by collecting the plantar inflammation site 3 hours after administration with the greatest inflammatory effect of carrageenan, using the ELISA kit from R&D System, and testing according to the manual of the vendor.

I. Experiment result

The experimental results of IL-1βIL-6 and PGE2 are shown in Table 5 and Figures 2 to 4 below.

Measurement result value IL-1β (pg/g) IL-6 (pg/g) PGE2 (pg/mL) carrageenan +Normal Saline 2521 7315 1442.667 carrageenan + Indomrthacin 1456.67 2699 549.667 carrageenan + Sugar and
Amino acid-containing solution 980 2021.33 400.667 carrageenan + Contains sugar
Sap 1211.67 2361.667 484

As a result of the experiment, the sugar and amino acid sap and sugar-containing sap solutions have lower cytokine levels compared to the control group administered with the carrageenan solution, which is an inflammation-inducing substance, and lower cytokine levels than indomethacin, an inflammation-reducing substance.

Therefore, it can be confirmed that the sap composition according to the present invention has an excellent anti-inflammatory effect.

The infusion composition of the present invention is excellent in suppressing inflammation.

Claims (4)

delete Infusion solution composition for intravenous treatment for inflammation, which contains glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine and N-acetylneuraminic acid, and further contains amino acids. . The method of claim 2, wherein the amino acid is isoleucine, leucine, lysine acetate, methionine, phenylalanine, threonine, tryptophan, valine, alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, proline, serine, tyrosine, and glycine Infusion composition characterized in that one or more selected. The composition of claim 2, wherein the amino acid is at least one of 20 amino acids capable of constituting a protein.

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