KR20060006353A - Composition for lowering blood glucose comprising hexose monophosphate - Google Patents

Abstract

The present invention relates to a composition comprising hexose monophosphate that orally administered hexose monophosphate increases the amount of pancreatic beta-cell insulin and lowers blood sugar by increasing the concentration of insulin in the blood.

Hexose monophosphate, glucose-1-phosphate, fructose-6-phosphate, hypoglycemic, insulin

Description

Composition for lowering blood sugar comprising hexose monophosphate {Composition for lowering blood glucose comprising hexose monophosphate}             

1 is a graph showing the change in blood glucose by oral administration of D-glucose-alpha-1-phosphate in the oral D-glucose loading test system of normal mice (VC is a control, ●: 30 mg / kg, ▲: 100 mg / kg, ▼: 300 mg / kg dose).

Figure 2 is a diagram showing the hypoglycemic effect of D-glucose-alpha-1-phosphate in streptozotocin-induced diabetic model mice (a: normal group, b: administration of streptozotocin only, c: streptozotocin + sterile distilled water) , d: 30 mg / kg, e: 100 mg / kg, f: 300 mg / kg, 1 to 8 represent mouse numbers).

3 is a bar graph measuring serum insulin concentrations of streptozotocin-induced diabetic model mice by administration of D-glucose-alpha-1-phosphate or D-fructose-6-phosphate.

4 is a diagram showing the effect of inhibiting pancreatitis and insulin secretion in streptozotocin-induced diabetic mice by administration of D-glucose-alpha-1-phosphate (control: normal group, STZ: streptozotocin only) Administration, vehicle control: streptozotocin plus sterile distilled water).

5 is a graph showing the last day blood glucose levels by repeated oral administration of D-glucose-alpha-1-phosphate in type 2 diabetes model mice (db / db).

6 is a graph showing the hypoglycemic effect of D-glucose-alpha-1-phosphate by oral glucose loading test system in type 2 diabetic model mice (db / db) (vehicle control: control, ●: 30 mg / kg, ▲: 100 mg / kg, ▼: 300 mg / kg dose).

7 is a graph showing the change in blood glucose by oral administration of D-fructose-6-phosphate in the oral glucose loading test system of normal mice (VC is a control, ●: 30 mg / kg, ▲: 100 mg / kg, ▼: 300 mg / kg administration).

8 is a graph showing the hypoglycemic effect of D-fructose-6-phosphate in streptozotocin-induced diabetic model mice (a: normal group, b: administration of streptozotocin only, c: streptozotocin + sterile distilled water) d: 30 mg / kg, e: 100 mg / kg, f: 300 mg / kg, 1 to 8 represent mouse numbers).

9 is a diagram showing the effect of inhibiting pancreatitis and insulin secretion of streptozotocin-induced diabetic model mice by the administration of D-fructose-6-phosphate (control: normal group, STZ: streptozotocin) Only administration, vehicle control: streptozotocin + sterile distilled water).

Figure 10 is a bar graph showing the last day blood glucose levels by repeated oral administration of D-fructose-6-phosphate in type 2 diabetes model mice (db / db).

FIG. 11 is a graph showing the hypoglycemic effect of D-fructose-6-phosphate by oral glucose loading on type 2 diabetic model mice (db / db) (vehicle control: control, ●: 30 mg / kg) , ▲: 100 mg / kg, ▼: 300 mg / kg administration).

The present invention relates to a blood sugar lowering composition comprising hexose monophosphate, a derivative thereof or a salt thereof.

Diabetes mellitus (DM) is a complex chronic disorder of carbohydrate, fat, and protein metabolism, mainly due to partial or complete lack of insulin by the pancreatic beta cells or lack of cellular insulin receptors. In normal people, hormones called insulin and glucagon are made in vivo to complement and lower or increase blood sugar to maintain proper blood glucose levels in vivo. Insulin is produced by beta-cells and stored, and when the blood sugar rises, insulin is secreted into the blood. The secreted insulin then enters the cells by binding to insulin receptors on the surface of muscle cells or hepatocytes, and then D- Glucose is introduced into the body and glucose metabolism is activated.

In Korea, diabetic patients accounted for 0.5% of the total population before the 1970s, so there was little interest in the medical community, but in the 1980s, 2-3%, and in the 1990s, 4-6% of the total population (about 1.5-2 million) Are estimated to be diabetic, and a significant number of people live without knowing that they have diabetes.

Symptoms of diabetes are diverse, but the most common ones are polyuria, Daum, and Dasik. In the case of diabetics, glucose is not used as an energy source, and thus the already stored protein and fat are consumed as energy sources. This results in weight loss.

Diabetes is largely divided into insulin-dependent diabetes (type 1 diabetes) and insulin-independent diabetes (type 2 diabetes). Type 1 diabetes is a condition in which the pancreatic β-cells are depleted due to genetic causes, viral infections, etc., and thus little insulin is secreted. It occurs mainly in the 10s and 20s, and is juvenile-onset. It is known as brittle or ketosis diabetes. Type 2 diabetes is unclear as to the cause of the disease, but due to the family history of diabetes, obesity, stress, etc. are well seen after 40s. In type 2 diabetes, the insulin secretion in the pancreas is sufficient but blood glucose is not normalized despite hyperinsulinemia due to insulin resistance and glucose utilization not being normal, and maturity development, adult development, ketosis resistance or stable diabetes It is said to be.

Diabetes is not a big disease in itself, but a complication that develops when the treatment of diabetes is delayed and becomes chronic, for example diabetic retinopathy (causes blindness, blindness, retinal hemorrhage, retinopathy and cataracts), diabetic nephropathy Peripheral neurosis, heart and circulatory disorders (caused angiopathy), periodontitis, bone deficiency, skin diseases, and the like are problematic. These pathological complications of diabetes are known to be fundamentally proportional to hyperglycemia (Porte, Jr. et al., 1996).

Currently used diabetic drugs are roughly divided into oral hypoglycemic agents and insulin injections. In general, insulin injection is given to insulin-dependent diabetic patients who do not have insulin secretion in the body, gestational diabetic patients, or insulin-independent diabetes patients whose blood glucose control is poorly controlled by oral hypoglycemic agents. It is common practice to take oral hypoglycemic drugs for non-controlled diabetic patients.

Insulin injections are usually given intravenously or intramuscularly, but subcutaneous injections are most common when long-term administration is required. Subcutaneous injection does not increase insulin rapidly and insulin secretion is reduced when ingesting food, and the effect of insulin is decreased by peripheral circulation rather than liver circulation (Goodman et al. The Pharmacological basis of Therapeutics, p1692). In addition, when genetically engineered insulin is injected, antibodies to insulin occur in the body due to structural defects, and in the case of patients receiving insulin for a long time, the administration of insulin continuously occurs because the generated antibodies neutralize insulin. There is a problem to increase the amount. Attempts have been made to develop insulin for oral administration, but insulin is structurally unstable, degrades in the gastrointestinal tract, and has a high molecular weight, resulting in low absorption of the peptide, which is still in experimental stage.

Commonly used oral hypoglycemic agents may be classified into sulfonylurea drugs, biguanide drugs, and alpha-glucosidase inhibitors (Deruiter's Endocrine Pharmacology Module, Spring, 2003). Sulfonylurea-based drugs include glypizide, glyclazide, glyquidone, glybenclamide, chlorpropamide, and the like, which promote insulin secretion in the pancreas. Therefore, they cannot be used in insulin-dependent diabetics who have no insulin secretion in the pancreas, and are used in patients with insulin-independent diabetes who have relatively reduced insulin secretion in the pancreas but are concerned about birth defects, abortion, and stillbirth. Therefore, there is a disadvantage that can not be used for women of childbearing age. In addition, these drugs can cause hypoglycemia when administered overdose or on an empty stomach, and also have side effects such as skin rash, jaundice, anorexia, nausea (nausea) and diarrhea. Biguanide-based drugs include metformin and the like, and have a weaker blood glucose lowering effect than sulfonylurea-based drugs, and are less likely to cause hypoglycemia. However, due to the high incidence of side effects of the digestive system, nausea, vomiting, diarrhea and rash appear at the beginning of treatment, and cause life-threatening side effects by causing lactic acidosis. have.

As you can see, today's methods for controlling diabetes are limited to only a few methods, and their approaches have a negative effect on the human body, unlike their natural glycemic control. It is necessary to develop drugs to replace.

The present invention relates to a blood sugar lowering composition comprising hexose monophosphate, a derivative thereof or a salt thereof.

The present invention provides a composition for lowering blood sugar, comprising hexose monophosphate, a derivative thereof, or a salt thereof.

The term "hexose monophosphate" used in the present invention is a sugar having a phosphate group at one position of carbon of hexose, and includes sugars that are present in nature or synthetically produced sugars.

The hexose constituting the hexose monophosphate of the present invention includes sugars having 6 carbon atoms or derivatives thereof, for example, glucose, fructose, galactose, gulose, Rhamnose, mannose, sorbose, allose, altrose, adiose, tagatose, talose, fucose or derivatives thereof And the like, and preferably glucose or fructose.

In the case of saccharides having one or more asymmetric carbons, there are D or L forms according to the absolute arrangement of the asymmetric carbon atoms furthest from the aldehyde or ketone group, and the hexose sugar of the present invention can be D or L form, preferably D form. .

Monophosphoric acid constituting the hexose monophosphate of the present invention refers to phosphate bonded to one position of the carbon hexose (phosphate), it is possible to bind to any one carbon of positions 1 to 6, preferably 1 Position 2, 6 or 6.

Examples of the hexose monophosphate of the present invention include D-glucose-alpha-1-phosphate, D-glucose-beta-1-phosphate, D-glucose-6-phosphate, D-brown sugar-1-phosphate, and D-brown sugar- 6-phosphate, D-mannose-alpha-1-phosphate, D-mannose-6-phosphate, D-allose-1-phosphate, D-alose-6-phosphate (D-allose -6-phosphate), D-altrose-1-phosphate, D-altrose-6-phosphate, D-ramrose-1-phosphate ( D-rhamnose-1-phosphate), D-diose-1-phosphate, D-diose-6-phosphate, D-fudose-6-phosphate (D-fucose-6-phosphate), D-gulose-1-phosphate, D-gulose-6-phosphate, L-Solbo-2 L-sorbose-2-phosphate, D-tagatose-2-phosphate, D-talose-1-phosphate, D-desugar -6-phosphate (D-talose-6-phosphate) or salts thereof, and the like.

The hexose sugar derivative of the present invention includes an unsubstituted or substituted amino hexose sugar or deoxygenated hexose sugar, and the amino hexose sugar includes amine glucose, glucosamine, galactosamine, mannosamine and the like. D-amine glucose-1-phosphate, D-amine glucose-6-phosphate, N-acetyl-D-amine glucose-6-phosphate, D-galactosamine-1-phosphate , D-galactosamine-6-phosphate, N-acetyl-D-galactosamine-1-phosphate, N-acetyl-D N-acetyl-D-galactosamine-6-phosphate, D-galactosamine-1-phosphate, D-amine-manganese-6-phosphate galactosamine-6-phosphate), N-acetyl-D-amine-mannose-1-phosphate, N-acetyl-D-amine-mandose-6-phosphate D-galactosamine-6-phosphate) or salts thereof, and the like.

Deoxygenated hexose sugar also refers to a sugar in which at least one hydroxy group of hexose carbon is replaced with hydrogen, for example, 2-deoxyoxy-D-glucose-1-phosphate (2-deoxy-D-glucose- 1-phosphate), 3-deoxygen-D-glucose-1-phosphate, 4-deoxygen-D-glucose-1-phosphate, 6-deoxygen-D-glucose-1-phosphate, 2,3-double Oxygen-D-glucose-1-phosphate, 2,4-double deoxygenation-D-glucose-1-phosphate, 2,6-double deoxygenation-D-glucose-1-phosphate, 3,4-double deoxygenation- D-glucose-1-1-phosphate, 3,6-double deoxygenation-D-glucose-1-phosphate, 4,6-double deoxygenation-D-glucose-1-phosphate, 1-deoxygenation-D-fructose -6-phosphate, 3-deoxygen-D-fructose-6-phosphate, 4-deoxygen-D-fructose-6-phosphate, 1,3-double-deoxygen-D-fructose-6-phosphate, 1,4 Double deoxygenation-D-fructose-6-phosphate, 3,4-double deoxygenation-D-fructose-6-phosphate or salts thereof.

The carbon of the hexose sugar may be substituted with a C ₁ to C ₆ straight or branched alkyl group, for example, the carbon at positions 1 to 6 is methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, One or more may be substituted with isopentyl, hexyl, isohexyl, heptyl, isoheptyl and the like.

The composition containing hexose monophosphate of the present invention may include one kind of hexose monophosphate or a mixture of several kinds thereof.

Hexose monophosphate of the present invention may be prepared in the form of a salt to which a cation is bound. The cation is sodium salt, potassium salt, magnesium salt, calcium salt, lithium salt, rubidium salt, cesium salt, belium salt, strontium salt, barium salt, aluminum salt, boron salt, gallium salt, copper salt, silver salt, zinc salt, Cations such as cadmium salts, mercury salts, scandium salts, nickel salts, manganese salts, chromium salts, barium salts, titanium salts.                     

The hexose monophosphate of the present invention can be preserved in itself or in an aqueous solution, and is chemically stable as an oxide of carbohydrates, so that there is little concern about degradation or alteration than insulin, which is a peptide even when stored for a long time.

In a preferred embodiment, the present invention provides a hypoglycemic composition comprising glucose monophosphate or fructose monophosphate, derivatives thereof, salts thereof. The composition of the present invention may comprise a single or a mixture of glucose monophosphoric acid or fructose monophosphate.

In a more preferred embodiment, the present invention provides a hypoglycemic composition comprising D-glucose-alpha-1-phosphate or D-fructose-6-phosphate, derivatives thereof, salts thereof.

Alpha-D-glucose-1-phosphate or D-fructose-6-phosphate used in the present invention can be obtained by synthesis from enzymes using starch, glucose or fructose as a raw material or purchased from a manufacturer.

D-glucose-alpha-1-phosphate is generally used to convert alpha-glucan (linear amylose, dextrin, starch, amylopectin, glycogen, etc.) in the presence of inorganic phosphoric acid (orthophosphoric acid, orthophosphate) to alpha-glucan phosphorylase (α-glucan). It can be synthesized through the enzymatic reaction of phospholyase). The enzyme reaction is preferably performed at a high temperature to reduce microbial infection in the sample and to increase the solubility of starch, which is a substrate, and for this, alpha-glucan phospho derived from thermophilic bacteria (eg, Thermus caldophilus GK24) that can react at 70 ° C. Preference is given to treating lilase.

In addition, D-fructose-6-phosphate first adds alpha-glucan phosphorylase to alpha-glucan (starch, glycogen, amylopectin, etc.) to produce alpha-D-glucose-alpha-1-phosphate, followed by glucose phosphate variation. The enzyme can be used to synthesize D-glucose-6-phosphate and finally produced via glucose-6-phosphate isomerase. Alternatively, it is possible to synthesize fructose by fructose kinase in the presence of ATP. Enzymes used in the production of D-fructose-6-phosphate are also needed at high temperatures to prevent contamination of the reactor from microorganisms at large scale and to increase the solubility of substrates in polymer form such as starch. It is preferred that it is a heat-resistant enzyme that acts.

The reaction product may be separated from impurities using chromatography or other methods known in the art.

The present invention relates to a method for enhancing production and secretion of insulin and lowering blood sugar with a composition comprising hexose monophosphate, a derivative thereof or a salt thereof.

In order for a drug to function in a living body, it must be absorbed into the body, and in the gastrointestinal system, the absorption of the drug depends on the absorption surface area, the blood flow of the absorption site, and the physical state of the drug formulation or solubility. Since the drug is passively absorbed at this site, the drug is nonionic and the higher the fat soluble, the higher the absorption. However, the hexose monophosphate of the present invention has a feature of exerting a biological function such as being absorbed into the body upon oral administration due to the phosphate group and promoting blood sugar drop and insulin secretion. Glycolic metabolites with two phosphate groups, such as D-fructose-1,6-double acid, appear to be difficult to absorb in vivo because they do not affect blood sugar control, but D-glucose-alpha-1-phosphate and D-fructose -6-Phosphate shows significant biofunctional control such as insulin production and secretion of pancreas, increase of insulin concentration in blood and hypoglycemia, so that hemosaccharide monophosphate can be infused orally by oral. .

The composition of the present invention serves to regulate blood glucose by activating the body's own glucose metabolism control function. In the case of administration of D-glucose-alpha-1-phosphate and D-fructose-6-phosphate, insulin is continuously produced in beta-cells for several days in streptozotocin-induced diabetic mice. It can be inferred that phosphoric acid and D-fructose-6-phosphate act as signaling agents to increase the expression of the insulin gene. Therefore, hexose monophosphate, such as D-glucose-alpha-1-phosphate and D-fructose-6-phosphate of the present invention, causes the pancreas to increase the expression of insulin genes in beta-cells and to properly secrete it into the blood. Blood glucose is regulated in a manner similar to the body's glucose metabolism control function, in which the concentration is increased and thus the blood glucose level is decreased.

As a specific example, oral administration of a composition comprising D-glucose-alpha-1-phosphate or D-fructose-6-phosphate to normal mice, type 1 diabetes mice and type 2 diabetes model mice (db / db) Results In all cases, insulin production of pancreatic beta-cells increased, followed by a decrease in blood glucose with high insulin levels in the blood.

Type 1 diabetic mice can be induced by administering to mice mice streptozotocin, a substance that can selectively destroy beta cells of the pancreas. In an embodiment of the present invention, when D-glucose-alpha-1-phosphate and D-fructose-6-phosphate are respectively administered to streptozotocin-induced diabetic mice, both have a hypoglycemic effect, and a small amount is administered. In one case, D-fructose-6-phosphate had a good suppression of blood glucose elevation, but when the dose was large, it was found that both had similar suppression of blood glucose rise or that D-glucose-alpha-1-phosphate was rather good (Example 2 and Example 5).

In addition, blood samples of streptozotocin-induced diabetic mice treated with hexose monophosphate were measured for insulin concentration. As a result, serum insulin was increased in both D-glucose-alpha-1-phosphate and D-fructose-6-phosphate. The concentration of serum insulin increased proportionally with the dose of monophosphate hexose, thereby lowering the level of blood sugar drop to the same level (Examples 2 and 5). D-fructose-6-phosphate tends to have a higher rate of increase in serum insulin and higher glucose-lowering ability than D-glucose-alpha-1-phosphate. Therefore, it can be seen that hexose monophosphate, such as D-glucose-alpha-1-phosphate and D-fructose-6-phosphate, can be used as an alternative medicine for insulin in diabetics.

After administration of the composition of the present invention, only insulin was selectively stained in pancreatic tissue samples of normal mice and streptozotocin-induced diabetic mice, and the amount of glucose drop in the pancreatic beta cells was observed under a microscope. Consistent with the increase in insulin concentrations in blood, it was confirmed that the distribution of beta-cell populations containing insulin in the pancreas was increased. Thus, the hexose monophosphate composition of the present invention promotes insulin production and secretion by the pancreatic beta-cells.

The composition of the present invention has a hypoglycemic effect even in type 2 diabetes. For example, model mice with type 2 diabetes (db / db) were treated with oral administration of D-glucose-alpha-1-phosphate and D-fructose-6-phosphate for 2 weeks or both. It was found that the blood glucose was strong (Examples 3 and 6).

As described above, for the purpose of lowering blood sugar, the composition including hexose monophosphate of the present invention is applicable to type 1 diabetic patients as well as type 2 diabetic patients.

Accordingly, the present invention provides a method of treating diabetes and related symptoms thereof with the compositions of the present invention. Diabetes and its related symptoms include insulin dependent diabetes mellitus (type 1 diabetes), insulin independent diabetes mellitus (type 2 diabetes), insulin resistance, hyperinsulinemia and hypertension caused by diabetes. Other diabetes-related symptoms include obesity and damage to blood vessels, eyes, kidneys, nerves, autonomic nervous system, skin, connective tissue, reproductive system and immune system.

As used herein, the term “treatment” means preventing, inhibiting and alleviating diabetes or related symptoms thereof. The composition of the present invention may itself be administered in a therapeutic dose or with a therapeutic dose of insulin for the purpose of lowering blood sugar. Or other hypoglycemic agents to treat damage to type 2 diabetes, insulin resistance, hyperinsulinemia, hypertension caused by diabetes, obesity, blood vessels, eyes, kidneys, nerves, autonomic nervous system, skin, connective tissue and the immune system. May be administered together.

As used herein, the term “therapeutic dose” may be a dose necessary to prevent diabetes or its related symptoms or may be a dose necessary to treat or alleviate the disease. In general, when administering a composition of the present invention together with insulin and / or a blood glucose lowering agent, the insulin and / or blood glucose lowering agent may be administered at a lower dose than when administered alone. Low doses of insulin and / or hypoglycemic agents can reduce the side effects of these drugs and delay the induction of complications resulting from diabetes or its related diseases.

Glucose lowering agents that can be used with the compositions of the present invention include metformin, acarbose, acetohexamide, glymepyride, tolazamide, glyphide, glyburide, tolbutamide, chloropropamide, thiazolidinedione, Alpha-glucose inhibitors, biguanide derivatives, troglitazones and mixtures thereof.

The composition of the present invention may be preferably administered in the form of injections such as oral, aerosol sprays, ointments, subcutaneous, muscle, intravenous, etc. For this purpose, tablets, capsules, powders, granules, microgranules, microspheres, suspensions, emulsion syrups Or in unit dosage forms or multiple dosage forms of pharmaceutical preparations, such as injections and the like. The composition of the present invention can be absorbed even during oral administration, and can also be preferably administered orally for ease of administration, in which case it can overcome the inconvenience and side effects of continuous insulin injection.

To prepare pharmaceutical formulations, the compositions of the present invention may be mixed with one or more inert, non-toxic, solid or liquid pharmaceutical carriers. As the pharmaceutical carrier, physiological saline, Ringer's solution, phosphate buffered saline and the like can be used. Pharmaceutical compositions may also contain stabilizers, antioxidants, excipients, dyes, binders, precipitants, dispersants, diluents, surfactants, preservatives, and the like. For example, inorganic substances such as calcium carbonate, magnesium carbonate, tricalcium phosphate, magnesium phosphate, kaolin, talc, magnesium stearate, silicon dioxide, titanium dioxide, zirconium dioxide or colloidal silica can be used, cellulose and its derivatives, Alginates, carrageates, chitosan derivatives, vegetable rubbers such as tragacanth rubber, guar rubber and derivatives thereof, organics of xane, rubber, starch, maltodextrin and vegetable oil can be used.

Alternatively, the composition of the present invention may be prepared by ingesting a solid or liquid food or prepared in cosmetic form.

The daily dosage of monophosphoric acid per hexane of the present invention may be administered in an amount of 30 to 300 mg per kg of body weight in mice and 10 to 100 mg per kg of body weight in humans. Can be. Dosages for a particular patient may vary depending on the patient's weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion, severity of the disease, and the like. The composition containing hexose monophosphate of the present invention showed no acute toxicity at all even when administered orally in large amounts in mice. Since the composition of the present invention does not produce antibodies to the body even when administered in the body, it is possible to overcome the problem of insulin resistance due to antibodies that may occur when administering insulin synthesized genetically or biochemically synthesized.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are merely to aid the understanding of the present invention, and the scope of the present invention is not limited to the following examples.

Example 1 . Measurement of the hypoglycemic effect of D-glucose-alpha-1-phosphate in the oral D-glucose load test.

Oral D-glucose load tester is an experimental system that measures insulin secretion ability and insulin action by elevated blood sugar, and comprehensively predicts the function of pancreatic beta cells and D-glucose absorption in target cells. .

Normal mice (ICR, Daehan Biolink Co., Ltd.) used about 22 g of 6-week-old females, and 6 animals per experimental group were fasted for 16 hours. D-glucose-alpha-1-phosphate (Sigma Co.) was prepared in sterile distilled water at a concentration of 30 mg / ml, diluted and orally administered to mice at concentrations of 30, 100 and 300 mg / kg (body weight), respectively. Administered. After 90 minutes, 2 g / kg of D-glucose dissolved in saline was orally administered to the mice in a liquid amount of 10 ml / kg. Blood glucose was measured immediately prior to dosing D-glucose (at 0 hours) and at 15, 60 and 120 minutes after dosing. Blood for blood glucose measurement was collected from the orbital vein of mice using a capillary tube. The collected blood was measured for blood glucose using Acculand Alpha (Boehringer mannheim Co.). The control group was administered sterile distilled water in the same amount instead of D-glucose-alpha-1-phosphate.

In all experimental groups, the administration of D-glucose increased blood glucose levels to a peak at 15 minutes and then decreased to a normal level after 120 minutes. 15 minutes after the highest blood glucose level, the control group increased 261.5 mg / dl compared to the blood glucose level before D-glucose administration. However, in the 30, 100 and 300 mg / kg group treated with D-glucose-alpha-1-phosphate. Inhibition of blood glucose increase was 38.6 (p <0.01), 37.2% and 29.3% (p <0.05), respectively, compared to the control group. These results indicate that D-glucose-alpha-1-phosphate has a hypoglycemic effect by oral administration (Table 1, Fig. 1).

* p <0.05, ** p <0.01

^a % inhibition = [(sample / solvent control group) increase in blood glucose x 100]-100

G1P: D-glucose-alpha-1-phosphate

Example 2 . Measurement of D-glucose-alpha-1-phosphate Lowering Glucose and Insulin Levels in Streptozotocin-induced Diabetic Test Systems

Streptozotocin (streptozotocin) is a substance that can cause insulin-dependent diabetes and is known to selectively destroy pancreatic beta cells. Streptozotocin (Sigma Co.) was used by dissolving in citric acid buffer (citrate buffer, pH4.2) just before use. Streptozotocin was induced three times through the abdominal cavity in mice (ICR) at a concentration of 50 mg / kg. Eight mice were administered to each experimental group and the experimental values were averaged.

On the 11th day, the blood glucose level was 175.3 mg / dl in normal mice without streptozotocin, but the blood glucose level increased to 219.0 mg / dl compared to day 0 in mice treated with streptozotocin and sterile distilled water (ICR). It can be seen that diabetes was induced (Table 2).

Blood glucose levels of 237.8, 174.2 and 82.7 mg / dl, respectively, when streptozotocin (50 mg / kg, three intraperitoneal administrations) and 30, 100 and 300 mg / kg of D-glucose-alpha-1-phosphate were simultaneously administered Showed an increase. In the case of administration of D-glucose-alpha-1-phosphate at the concentration of 30 mg / kg, the effect of suppressing blood glucose elevation was not observed, but in the group of 100 and 300 mg / kg, 20.5 compared to the control group treated with streptozotocin and sterile distilled water. Inhibition of blood glucose elevation was shown in% and 62.2% (Table 2, Figure 2).

* p <0.05

^a % inhibition = [(sample / control) blood glucose increase x 100]-100

STZ: streptozotocin

G1P: D-glucose-alpha-1-phosphate

In addition, all mice in the experimental group were examined on day 11, and serum was isolated, and then the amount of insulin in the blood was measured using a mouse insulin elisa kit (ELISA Kit, Shibayagi, Japan). In other words, the anti-insulin coated plate was washed four times with washing buffer (Shibayagi buffer), followed by 100 uL of biotin conjugated anti-insulin and 10 uL of plasma sample. To each well of the anti-insulin test plate (well) and left for 2 hours at room temperature. After washing four times with washing buffer (I), 100 ul of streptavidin solution (HRP conjugated streptavidin solution) was added to each test tube and allowed to stand at room temperature for 30 minutes. The test plate was washed four times with a wash buffer, and then 100 uL of substrate chromogen reagent was added and reacted at room temperature for 30 minutes. The reaction was stopped by adding 100 uL of reaction stopper. When the absorbance of each test cell was measured at 450 nm using a precision microplaste reader (Molecular Devices, USA), 300 mg / kg of D-glucose-alpha-1-phosphate was coadministered with streptozotocin. , It can be seen that the amount of serum insulin is up to 6 times higher than that of streptozotocin and annihilated distilled water (Table 3, Figure 3).

* STZ: streptozotocin

On the other hand, the histopathological examination was performed to investigate the inhibitory effect of pancreatitis induced by streptozotocin-induced beta-cell destruction and the pancreatic insulin production and / or secretion effect. In insulin-dependent diabetes, pancreatitis is caused by immune cells, such as macrophages and T cells, invading the pancreas and destroying beta cells.

Pancreas present between duodenum and spleen after sacrifice of mice treated with D-glucose-alpha-1-phosphate to observe pancreatitis inhibitory effect and pancreatic beta-cell insulin production by the administration of hexose monophosphate Was removed with surgical scissors and fixed in 10% neutral formalin, and then embedded in paraffin according to the usual method, and then cut into a thickness of 5 um with an ultra-thin cutting machine to make a tissue specimen. The anti-human guinea pig using ABC kit (Vector) Insulin immunostaining was performed with insulin antibodies.

The pathologic findings showed that the number of beta cells positive for insulin was significantly decreased in the group treated with streptozotocin alone, whereas the group positive in proportion to the dose was administered in the group treated with D-glucose-alpha-1-phosphate. Increasing the number of cells and staining of the stain, it was shown that not only the effect of inhibiting pancreatitis (insulitis) but also the effect of promoting insulin secretion (FIG. 4, 3 pictures per experimental group). From these results, it can be seen that D-glucose-alpha-1-phosphate effectively lowers blood glucose levels and promotes insulin production / secretion in diabetes caused by administration of streptozotocin.

Example 3 . Therapeutic Effect of Oral Administration of D-Glucose-alpha-1-phosphate in Type 2 Diabetes (Insulin-Independent Diabetes) Model db / db Mice

D-glucose-alpha-1-phosphate was orally administered to measure hypoglycemic effect in type 2 diabetes (insulin-independent diabetes) model db / db mice. Type 2 diabetes is caused by an increase in resistance to insulin in the peripheral tissues, such as obesity, resulting in hyperinsulinemia. Beta cells of type 2 diabetic patients continue to secrete insulin without recognizing the excretion of glucose into the urine due to decreased response to insulin secretion due to elevated blood glucose. Model mice exhibiting these symptoms (db / db) are the best genetic diabetes model animals for studying human insulin-independent diabetes.

Mice (db / db, 21-31 g, SLC Co.) were orally administered once daily at concentrations of 30, 100 and 300 mg / kg of D-glucose-alpha-1-phosphate for 2 weeks from 5.5 to 7.5 weeks of age. Afterwards, the amount of glucose in the blood was measured on the last day of administration.

The control model mice (db / db) who received the same amount of sterile distilled water instead of D-glucose-alpha-1-phosphate showed 700.8 mg / dl of blood glucose on the last day. On the other hand, in the model mice (db / db) of the experimental group administered with D-glucose-alpha-1-phosphate at concentrations of 30, 100 and 300 mg / kg, 711.1, 662.4 and 543.1 mg / dl, respectively, It can be seen that alpha-1-phosphate has a hypoglycemic effect in type 2 diabetes model mice (db / db) (Table 4, Figure 5).

G1P: D-glucose-alpha-1-phosphate

In addition, type 2 diabetes (insulin-independent diabetes) model mice (db / db) using the oral glucose tolerance test (Oral glucose tolerance test) in the same manner as in Example 1 was further confirmed the hypoglycemic effect.

D-glucose-alpha-1-phosphate (Sigma Co.) was orally administered to model mice (db / db) fasted for 16 hours at concentrations of 30, 100 and 300 mg / kg body weight. Thereafter, D-glucose was orally administered to the mice in a liquid amount of 10 ml / kg so that 2 g / kg was administered after 90 minutes. Blood glucose was measured immediately prior to dosing (0 hour) and at 15, 30, 60, 120 and 180 minutes after dosing D-glucose. The control group was orally administered sterile distilled water in the same amount instead of D-glucose-alpha-1-phosphate.

In all experimental groups, D-glucose administration resulted in an increase in blood glucose levels, which peaked after 15-30 minutes. In the case of the control group, the time to return to the initial blood glucose level after administration of D-glucose was about 180 minutes, and 180, 30, 100 and 300 mg / kg of D-glucose-alpha-1-phosphate, respectively, It took about 120 and 60 minutes (Table 5, Figure 6). These results indicate that D-glucose-alpha-1-phosphate has a strong hypoglycemic effect in type 2 diabetes model mice (db / db).

Example 4 Hypoglycemic Effects of D-Fructose-6-Phosphate in Oral D-Glucose Tolerance Test

Normal mice [ICR, Daehan Biolink Co., Ltd.] used about 22 g of 6-week-old females and fasted six animals per experimental group for 16 hours. D-fructose-6-phosphate (Sigma Co.) was prepared in sterile distilled water at a concentration of 30 mg / ml, diluted and orally administered to mice at concentrations of 30, 100 and 300 mg / kg (body weight), respectively. . After 90 minutes, 2 g / kg of D-glucose dissolved in saline was orally administered to the mice in a liquid amount of 10 ml / kg.                     

Blood glucose was measured immediately prior to dosing D-glucose (at 0 hours) and at 15, 60 and 120 minutes after dosing.

Blood for glucose measurement was collected from the orbital vein of mice using capillaries. The collected blood was measured for blood glucose using Acculand Alpha (Boehringer mannheim Co.). The control group was administered sterile distilled water in the same amount instead of D-fructose-6-phosphate.

In all experimental groups, D-glucose administration increased blood glucose levels to the highest level at 15 minutes, and then decreased to normal levels after 120 minutes. At 15 minutes, blood glucose levels peaked at 256.1 mg / dl in the control group compared to pre-dose glucose, but in the 30, 100 and 300 mg / kg groups treated with D-fructose-6-phosphate, In comparison, 25.0, 48.4% (p <0.001) and 66.5% (p <0.001) inhibition of blood sugar elevations were shown. These results indicate that D-fructose-6-phosphate has a hypoglycemic effect by oral administration (Table 6, Figure 7).

*** p <0.01

^a % inhibition = [(sample / control) blood glucose increase x 100]-100

F6P: D-Fructose-6-Phosphate

Example 5 . Effect of D-Fructose-6-Phosphate on Lowering Glucose and Increasing Blood Insulin in Streptozotocin-induced Diabetes Mellitus

In the same mice and methods as in Example 2, the effect of D-fructose-6-phosphate was measured on the hypoglycemic and blood insulin levels in the streptozotocin-induced diabetes test system.

In the mice without streptozotocin (ICR), the blood glucose level was 175.3 mg / dl on the last day, and the mice with the streptozotocin and sterile distilled water (ICR) showed an increase in blood glucose level of 219.0 mg / dl compared to the initial blood glucose level. . On the other hand, sterile distilled water was administered at 203.1, 176.0, and 89.2 mg / dl, respectively, in the model mice (db / db) in which the streptozotocin and D-fructose-6-phosphate were administered at concentrations of 30, 100, and 300 mg / kg. Inhibition of 7.3%, 19.6% and 59.3% compared to one solvent control group shows that D-fructose-6-phosphate has a hypoglycemic effect in type 2 diabetes model mice (db / db) (Table 7 8). Compared to the case of administering D-glucose-alpha-1-phosphate of Example 2, the lower the blood glucose suppression rate of D-fructose-6-phosphate when the dose is administered, but the blood glucose inhibition rate is similar when the dose is high Or D-glucose-alpha-l-phosphate.

** p <0.01

^a % inhibition = [(sample / solvent control group) increase in blood glucose x 100]-100

STZ: streptozotocin

F6P: D-Fructose-6-Phosphate

In addition, in the same manner as in Example 2, the mice of the experimental group were autopsied on day 11, and serum was separated, and then the amount of insulin in the blood was measured using a mouse insulin elisa kit (ELISA Kit, SHABAYAGI, Japan). When 300 mg / kg of D-fructose-6-phosphate is administered with streptozotocin, the amount of insulin in the blood is up to 9 times higher than that of streptozotocin and sterile distilled water. At this time, the amount of insulin in blood was higher than that of the normal group mice (Table 8, Figure 3). As a result, it can be seen that administration of D-fructose-6-phosphate is better for the increase rate of serum insulin and hypoglycemic effect than D-glucose-alpha-1-phosphate of Example 2.

* STZ: streptozotocin

On the other hand, after oral administration of D-fructose-6-phosphate, histopathological examination was performed to inhibit pancreatitis (insulitis) induced by the destruction of beta-cells by streptozotocin and to promote insulin production and / or secretion. Learned about.

After administering D-fructose-6-phosphate in the same manner as in Example 2, the pancreas of the mice were isolated, fixed with formalin, and tissue samples were prepared for insulin immunostaining.

Pathologically, the number of beta cells positive for insulin was significantly decreased in the group treated with streptozotocin alone, whereas the number of insulin-positive cells in the group treated with D-fructose-6-phosphate was proportional to the dose. Increasing the staining and staining of the dye, as well as the inhibitory effect of pancreatitis (insulitis), especially insulin secretion promoting effect was shown to be strong (Fig. 9, three photos per experimental group). From these results, it can be seen that D-fructose-6-phosphate effectively lowers blood glucose levels and promotes insulin production / secretion in diabetes induced by administration of streptozotocin.

Example 6 Therapeutic Effect by Oral Administration of D-Fructose-6-Phosphate in Type 2 Diabetes (Insulin-Independent Diabetes) Model Mice (db / db)

D-fructose-6-phosphate was orally administered in the same manner as in Example 3 to determine the effect of hypoglycemia in type 2 diabetes (insulin-independent diabetes) model db / db mice.

The control model mice (db / db) who received the same amount of sterile distilled water instead of D-fructose-6-phosphate showed 700.8 mg / dl of blood sugar at the last day. On the other hand, the model mice (db / db) of the experimental group administered D-fructose-6-phosphate at concentrations of 30, 100 and 300 mg / kg showed 663.1, 611.6 and 603.0 mg / dl, respectively. It can be seen that 1-phosphate has a hypoglycemic effect in type 2 diabetes model mice (db / db) (Table 9, FIG. 10).

F6P: D-Fructose-6-Phosphate

In addition, type 2 diabetes (insulin-independent diabetes) model db / db mice using the Oral glucose tolerance test (Oral glucose tolerance test) as in Example 1 was further confirmed the hypoglycemic effect.

D-fructose-6-phosphate was orally administered to model mice (db / db) fasted for 16 hours at concentrations of 30, 100 and 300 mg / kg body weight. D-glucose was orally administered to the mice in a liquid amount of 10 ml / kg so that 2 g / kg was administered after 90 minutes. Blood glucose was measured immediately prior to dosing (0 hour) and at 15, 30, 60, 120 and 180 minutes after dosing D-glucose. As a control, sterile distilled water was used instead of D-fructose-6-phosphate.

In all experimental groups, D-glucose administration resulted in an increase in blood glucose levels, which peaked after 15-30 minutes. In the case of control group administered sterile distilled water, the time required to return to the initial blood glucose level after D-glucose administration was 180 minutes, and 180, 30, 100 and 300 mg / kg of D-fructose-6-phosphate, respectively. It took about 180 and 120 minutes (Table 10, Figure 11). These results indicate that D-fructose-6-phosphate has a strong hypoglycemic effect in type 2 diabetes model mice (db / db).

F6P: D-Fructose-6-Phosphate

Compositions comprising hexose monophosphate, derivatives thereof or salts thereof of the present invention can lower blood sugar by increasing the amount of pancreatic beta-cell insulin and increasing the concentration of insulin in the blood.

Claims (14)

A composition for lowering blood sugar comprising hexose monophosphate, a derivative thereof, or a salt thereof. The composition according to claim 1, wherein the hexose sugar is glucose, fructose, brown sugar, oyster sugar, lamb's sugar, man sugar, solvo sugar, egg sugar, alt sugar, disaccharide, taga sugar, de-sugar or fu sugar. The composition of claim 1 wherein the hexose monophosphate is hexose-1-phosphate, hexose-2-phosphate or hexose-6-phosphate. The composition of claim 1 wherein the hexose monophosphate is glucose monophosphate or fructose monophosphate. The composition of claim 1, wherein at least one carbon of the hexose sugar is substituted with methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, hexyl, isohexyl, heptyl or isoheptyl groups. The salt according to claim 1, wherein the salt is sodium salt, potassium salt, magnesium salt, calcium salt, lithium salt, rubidium salt, cesium salt, belium salt, strontium salt, barium salt, aluminum salt, boron salt, gallium salt, copper salt , Silver salt, zinc salt, cadmium salt, mercury salt, scandium salt, nickel salt, manganese salt, chromium salt, Valium salt or titanium salt. The method of claim 1, wherein the hexose monophosphate is D-glucose-alpha-1-phosphate, D-glucose-beta-1-phosphate, D-glucose-6-phosphate, D-brown sugar-1-phosphate, D-brown sugar -6-phosphate, D-mannose-alpha-1-phosphate, D-mannose-6-phosphate, D-saccharide-1-phosphate, D-saccharide-6-phosphate, D-altsugar-1-phosphate, D- Alt sugar-6-phosphate, D-ram sugar-1-phosphate, D-disaccharide-1-phosphate, D-disaccharide-6-phosphate, D-fudang-6-phosphate, D-gulose-1-phosphate, D-gulose -6-phosphoric acid, L-Solbosaccharide-2-phosphate, D- tagasugar-2-phosphate, D-desugar-1-phosphate or D-desugar-6-phosphate. 8. The composition of claim 7, wherein the hexose monophosphate is D-glucose-alpha-1-phosphate. 8. The composition of claim 7, wherein the hexose monophosphate is D-fructose-6-phosphate. The composition of claim 1, wherein the derivative of hexose monophosphate is aminohexose monophosphate or deoxygenated hexose monophosphate. The derivative of hexose monophosphoric acid according to claim 10, wherein the derivative of hexose monophosphate is D-amine glucose-1-phosphate, D-amine glucose-6-phosphate, D-fructose-6-phosphate, D-fructose-2-phosphate, N-acetyl D-amineglucose-1-phosphate, N-acetyl-D-amineglucose-6-phosphate, D-amine brown sugar-1-phosphate, D-amine brown sugar-6-phosphate, N-acetyl-D-amine brown sugar- 1-phosphoric acid, N-acetyl-D-amine brown sugar-6-phosphate, D-amine man sugar-1-phosphoric acid, D-amine mandose-6-phosphoric acid, N-acetyl-D-amine mandose-1-phosphoric acid or N- The composition is acetyl-D-amine mannose-6-phosphate. The derivative of hexose monophosphoric acid according to claim 10, wherein the derivative of hexose monophosphoric acid is 2-deoxygen-D-glucose-1-phosphate and 3-deoxygen-D-glucose-1-phosphate, 4-deoxygen-D-glucose-1- Phosphoric Acid, 6-Deoxygen-D-Glucose-1-phosphate, 2,3-Double Deoxygen-D-Glucose-1-phosphate, 2,4-Double Oxygen-D-glucose-1-phosphate, 2,6 -Double Deoxygen-D-Glucose-1-Phosphate, 3,4-Double Deoxygen-D-Glucose-1-1-Phosphate, 3,6-Double Deoxygenation-D-Glucose-1-Phosphate, 4,6 -Double Deoxygen-D-Glucose-1-Phosphate, 1-Deoxygen-D-Fructose-6-Phosphate, 3-Deoxygen-D-Fructose-6-Phosphate, 4-Deoxygen-D-Fructose-6- Phosphoric Acid, 1,3-Dual Deoxygenation-D-Fructose-6-Phosphate, 1,4-Double Deoxygenation-D-Fructose-6-Phosphate or 3,4-Dual Deoxygenation-D-Fructose-6-Phosphorus Composition. The composition of claim 1 in the form of an oral, aerosol spray, ointment or injection. 13. A composition according to any one of the preceding claims in the form of a food or cosmetic in solid or liquid form.

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