CN112402445A - Application of chitosan oligosaccharide biguanide derivative in preparation of medicine for treating liver insulin resistance - Google Patents
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Abstract
The invention discloses an application of a chitosan oligosaccharide biguanide derivative in preparing a medicament for treating liver insulin resistance. In the invention, a type 2diabetes rat model is established, the chitosan oligosaccharide biguanide derivative is administrated in a gastric perfusion mode, and after the treatment of the derivative, the insulin level is increased, so that the sensitivity of peripheral tissues to insulin is increased; meanwhile, IRS-2 tryptophan site phosphorylation is activated, and then a PI3K-Akt insulin signal conduction pathway is activated, so that on one hand, the expression of downstream signal protein GLUT-2 is improved, and the glucose uptake of the liver is increased; on the other hand, the expression of PEPCK and G6Pase is inhibited, and then hepatic gluconeogenesis is inhibited, and finally, the therapeutic effect on hepatic insulin resistance is exerted.
Description
Technical Field
The invention belongs to the field of pharmacy, and particularly relates to a microwave synthesis preparation method of a chitosan oligosaccharide biguanide derivative and application of the chitosan oligosaccharide biguanide derivative in preparation of a medicament for treating liver islet deficiency.
Background
Insulin Resistance (IR) and pancreatic beta-cell injury are currently considered to be the pathogenesis of type 2diabetes (T2DM) as chronic metabolic diseases (Kim, E. -A., Lee, S. -H., Lee, J. -H., Kang, N., et al., A marine algal polyphenol, dieckol, intestinal blood glucose levels by Akt pathway in oxygenated diabetic hypertension induced hypertension model [ J ]. RSC Advances,2016,6, 78570-78575). The liver plays an important role in insulin resistance as a main organ of insulin action and energy balance in the body, and thus, intervening in liver insulin resistance can effectively prevent metabolic diseases.
It has been shown that insulin action in the liver is mainly manifested in the IRS/PI3K/Akt signaling pathway (Gao, Y. -f., Zhang, M. -n., Wang, T. -x., Wu, T. -c., et al., hyperglycemic effect of D-chiro-insulin in type 2diabetes mellitus series through the PI3K/Akt signaling pathway [ J ]. Molecular and cellular endocrinology,2016,433, 26-34). Insulin is the initiating factor of the signaling pathway, which upon binding to the hepatic insulin receptor phosphorylates the insulin substrate and activates the PI3K/Akt signaling pathway. PI3K is one of the key proteins mediating insulin resistance, which when activated mediates Akt activation and affects the expression of glucose transporter type 2 (GLUT-2) and glucose kinase (GCK) (Motoyoshi, S., Shirotani, T., Araki, E., Sakai, K., et al, Cellular characterization of structural adenosine line (AtT20cell) transformed with insulin, glucose transporter type 2(GLUT2) and glucose genes insulin secretion from physical uptake centers of glucose [ J ]. abenterology, 1998,41,1492 1501). Among them, GLUT-2 has no restriction effect on the entrance of glucose into liver, but is a key rate-limiting switch in the process of inputting and outputting blood to and from the liver of glucose (Cohen, M., Kitsberg, D., Tsytkin, S., Shulman, M., et al., Live imaging of GLUT2glucose-dependent influencing and inhibiting in polarized extracellular cultures [ J ]. Open biology,2014,4,140091), and after being regulated by upstream molecules, extracellular glucose can be transported to cells through GLUT-2 for storage, which can reduce blood glucose. Evidence suggests that normal insulin secretion is ensured only when GLUT-2 and GCK are normally expressed; in addition, activated Akt can promote phosphorylation of FoxO1 in the cytoplasm of hepatocytes, inhibit nuclear transfer, and cause inhibition of the interaction between the promoters of FoxO1 and phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase), and ultimately affect the expression of PEPCK and G6Pase, key enzymes for gluconeogenesis, thereby regulating hepatic gluconeogenesis (Kim, S.J., Quan, H.Y., Jeong, K.J., Kim, D.Y., et al, Beicia effect of beta acid on hyperglycemic vitamin a recycling of hepatic glucose product [ J ]. Journal of aggregate and food chemistry,2013,62, 434. 442). The above pathways act together to maintain the steady state of the liver blood sugar concentration, so that the intervention on the liver PI3K-Akt signal pathway has extremely important significance for effectively treating type 2 diabetes.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides the application of the chitosan oligosaccharide biguanide derivative in preparing the medicine for treating liver insulin resistance.
The chitosan oligosaccharide biguanide derivative used in the invention adopts the technical scheme of chitosan oligosaccharide biguanide derivative and microwave synthesis method thereof (application No. 2017108673840, application date 2017, 9 and 22 days), the microwave synthesis method is used for introducing the biguanide structure of the classic diabetes treatment medicament metformin into chitosan oligosaccharide, and the product has the effect of treating the liver injury of diabetes with guanidino, has the naturalness, safety and unique biological activity of chitosan oligosaccharide, and can relieve the stimulation side effect of biguanide medicament on gastrointestinal tract.
The chitosan oligosaccharide biguanide derivative is prepared by taking chitosan oligosaccharide and dicyandiamide as raw materials and taking chitosan oligosaccharide side group NH2And dicyandiamide inReacting under hydrochloric acid and microwave conditions to form a biguanide group bonded to the chitosan oligosaccharide, as shown in the following chemical formula:
the chitosan oligosaccharide is the only basic amino oligosaccharide with positive charges in nature, has small molecular weight, good solubility and easy absorption and utilization by organisms, the weight average molecular weight is below 3000, preferably 1500-3000 Da, and the deacetylation degree is above 97%, preferably 97-99%.
In the chitosan oligosaccharide biguanide derivative, the substitution degree of biguanide is 40-60%, preferably 45-55%.
In the chemical formula, m and n are respectively the polymerization degrees of acetylation units and deacetylation units in the chitosan oligosaccharide.
The preparation method of the chitosan oligosaccharide biguanide derivative comprises the following steps: dispersing chitosan oligosaccharide in hydrochloric acid and stirring under the microwave condition of 160W-560W to perform the first step of microwave reaction; adding dicyandiamide aqueous solution, adjusting the pH value of a reaction system to 1-2 by using hydrochloric acid, continuously stirring under the microwave condition of 160-560W, and carrying out a second-step microwave reaction to react chitosan oligosaccharide and dicyandiamide to prepare the chitosan oligosaccharide biguanide derivative, wherein the specific formula is shown as follows:
in the first step of microwave reaction, the microwave power is 300-400 w, the reaction time is 3-7 min, preferably 5-7 min, and the stirring speed is 100-200 rpm.
In the second step of microwave reaction, the microwave power is 300-400 w, the reaction time is 10-20 min, preferably 15-20 min, and the stirring speed is 100-200 rpm.
And after the reaction is finished, cooling the reaction liquid to room temperature, carrying out alcohol precipitation on the mixed solution, drying and grinding to obtain chitosan oligosaccharide biguanide derivative powder.
The molar ratio of dicyandiamide to amino groups in the chitosan oligosaccharide is (0.5-3): 1, preferably (1-3): 1, and more preferably 1: 1.
Hydrochloric acid is 0.2-0.8 mol/L HCl aqueous solution, and the pH value of the reaction system is adjusted to 1 by using the hydrochloric acid.
The invention has the advantages of high production efficiency, simple operation process, clean and environment-friendly reaction process and good product performance. Animal experiments prove that the chitosan oligosaccharide biguanide derivative is administered by intragastric administration after a type 2 diabetic rat model is established, and after the chitosan oligosaccharide biguanide derivative is treated by the derivative, the insulin level is increased, so that the sensitivity of peripheral tissues to insulin is increased; meanwhile, the phosphorylation of the position 473 of IRS-2 tryptophan is activated, so that a PI3K-Akt insulin signal conduction pathway is activated, on one hand, the expression of downstream signal proteins GLUT-2 and GCK is improved, and the glucose uptake of the liver is increased; on the other hand, the expression of PEPCK and G6Pase is inhibited, and the hepatic gluconeogenesis is further inhibited. The chitosan oligosaccharide biguanide derivative has better treatment effect compared with the traditional diabetes treatment medicament metformin which is used alone, namely the application of the chitosan oligosaccharide biguanide derivative in preparing the medicament for treating the liver insulin resistance.
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FIG. 1 is a comparison spectrum diagram of infrared spectra of raw materials Chitosan Oligosaccharide (COS) and chitosan oligosaccharide biguanide hydrochloride (COSG) in the invention.
FIG. 2 is a nuclear magnetic carbon spectrum contrast line chart of Chitosan Oligosaccharide (COS) and chitosan oligosaccharide biguanide hydrochloride (COSG) as raw materials in the invention.
FIG. 3 is a graph of data on the effect of COSG on STZ-induced body weight, fasting plasma glucose and urine glucose in STZ-induced diabetic rats, where (a) is body weight, (b) is fasting plasma glucose, (c) is urine glucose, the data are mean. + -. standard deviation, and n is 6,#P<0.05,##P<0.01,###P<0.001 vs. blank control group,. P<0.05,**P<0.01,***P<0.001 compared to the model group.
FIG. 4 is a graph of the data for the effect of COSG on the insulin (a) and HOMA-IR (b) of STZ-induced diabetic rats of the present invention, where the data are mean. + -. standard deviation, n is 6,#P<0.05,##P<0.01,###P<0.001 vs. blank control group,. P<0.05,**P<0.01,***P<0.001 compared to the model group.
FIG. 5 is a Western blot analysis of IRS-2(a), Akt and p-Akt (ser473) (b) in STZ-induced diabetic rats of the present invention; histogram of data for quantitative analysis of protein expression of IRS-2(c), Akt (d), p-Akt (ser473) (e) and p-Akt (ser473)/Akt (f) in STZ-induced diabetic rats, where the data are mean ± standard deviation, n ═ 6,#P<0.05,##P<0.01,###P<0.001 vs. blank control group,. P<0.05,**P<0.01,***P<0.001 compared to the model group.
FIG. 6 is a graph of data for Western blot analysis (a), quantitative analysis (b) of GLUT-2 protein expression, and quantitative analysis (c) of GCK expression of STZ-induced diabetic rats in accordance with the present invention, wherein the data are mean. + -. standard deviation, n is 6,#P<0.05,##P<0.01,###P<0.001 vs. blank control group,. P<0.05,**P<0.01,***P<0.001 compared to the model group.
FIG. 7 is a bar graph of the effect of COSG on PEPCK (a), G6Pase (b) and liver glycogen (c) in STZ-induced diabetic rats according to the present invention, wherein the data are mean. + -. standard deviation, n is 6,#P<0.05,##P<0.01,###P<0.001 vs. blank control group,. P<0.05,**P<0.01,***P<0.001 compared to the model group.
Detailed Description
The following is a further description of the invention and is not intended to limit the scope of the invention. After the chitosan oligosaccharide biguanide derivative is prepared, the subject group successively tests and evaluates the medicinal value of the chitosan oligosaccharide biguanide derivative, such as the application of the previous Chinese patent application of ' application of the chitosan oligosaccharide biguanide derivative in preparing a medicament for treating diabetic nephropathy ' (application No. 2017108663660, application No. 2017, 9 and 22 days), ' application of the chitosan oligosaccharide biguanide derivative in preparing a medicament for treating lipid metabolism disorder ' (application No. 2017108917900, application No. 2017, 9 and 27 days), ' application of the chitosan oligosaccharide biguanide derivative in preparing a medicament for treating pancreatic islet deficiency ' (application No. 2018111171793, application No. 2018, 9 and 25 days), ' application of the chitosan oligosaccharide biguanide derivative in preparing a medicament for inhibiting apoptosis (application No. 2018111202043, application No. 2018, 9 and 25 days), ' application of the chitosan oligosaccharide biguanide derivative in preparing a medicament for treating skeletal muscle insulin resistance ' (application No. 2018111927902, application date 2018, 10 month 13). In the present invention, this subject group continues this test for the medicinal value of chitosan oligosaccharide biguanide derivatives, which is to prepare and characterize chitosan oligosaccharide biguanide derivatives first and study them for insulin resistance.
Example 11500 Da preparation of Chitosan oligosaccharide biguanide derivatives
The raw material ratio is 1:1, weighing 1.44g of dicyandiamide in a conical flask, adding 15mL of distilled water, and shaking and dissolving dicyandiamide in a constant-temperature shaking incubator at 50 ℃. After the chitosan oligosaccharide (1500Da) and the dicyandiamide are completely dissolved, pouring the dicyandiamide aqueous solution into a four-mouth bottle containing the chitosan oligosaccharide hydrochloric acid solution, and adjusting the pH value of the solution to 1 by using 1mol/L hydrochloric acid. The reaction was carried out under conditions of microwave power of 400W and microwave time of 15 min. After the reaction is finished, carrying out alcohol precipitation, suction filtration, drying and grinding on the mixed solution to obtain chitosan oligosaccharide biguanide derivative powder COSG with the degree of substitution of 55%.
Example 23000 Da preparation of Chitosan oligosaccharide biguanide derivatives
The raw material ratio is 1:1 weighing 2.88g of dicyandiamide in a conical flask, adding 15mL of distilled water, and shaking and dissolving dicyandiamide in a constant-temperature shaking incubator at 50 ℃. After the chitosan oligosaccharide (3000Da) and the dicyandiamide are completely dissolved, pouring the dicyandiamide aqueous solution into a four-mouth bottle containing the chitosan oligosaccharide hydrochloric acid solution, and adjusting the pH value of the solution to 1 by using 1mol/L hydrochloric acid. The reaction was carried out under conditions of microwave power of 400W and microwave time of 15 min. After the reaction is finished, carrying out alcohol precipitation, suction filtration, drying and grinding on the mixed solution to obtain chitosan oligosaccharide biguanide derivative powder COSG with the degree of substitution of 55%.
The chitosan oligosaccharide biguanide derivative and the raw material chitosan oligosaccharide obtained in the embodiment 1 of the invention are identified by infrared spectroscopy, nuclear magnetic resonance spectroscopy and the like, as shown in attached figures 1 and 2.
In the infrared spectrum of COS, 3414cm-1O-H and N-H stretching vibration with strong absorption peaks near the vicinity of the vibration and the hydrogen bond associationDynamic absorption peaks are partially overlapped to widen multiple absorption peaks. Because a large number of intermolecular and intramolecular hydrogen bonds exist in COS molecules, and the lengths and the strengths of the hydrogen bonds are different, the stretching vibration peak of the COS molecules appears in a wider frequency range. 2918cm-1The left and right sides are C-H stretching vibration absorption peaks. 1626cm-1The vicinity is the N-H stretching vibration peak of secondary amine. Furthermore, 1060cm of the fingerprint area-1An N-H in-plane bending vibration absorption peak of secondary amine is close to the point, because COS deacetylation is not complete and a small amount of-NH-CO-structure is contained in the molecule, an absorption peak is formed at the point; 602cm-1The vicinity corresponds to the out-of-plane rocking vibration absorption band of N-H. Compared with COS, COSG 1626cm-1At 1060cm-1The absorption peak near the molecule is obviously enhanced, and both the peaks are characteristic peaks of secondary amine groups, which indicates that secondary amine groups are increased in the molecule, and indicates that the guanidination reaction is performed on COS amino groups. In addition, 3414cm-1The nearby absorption peak is a multiple absorption peak widened by partial overlapping of O-H and N-H stretching vibration absorption peak parts associated with hydrogen bonds, because a large number of intermolecular and intramolecular hydrogen bonds exist in COS molecules, and the length and the strength of the hydrogen bonds are different, the stretching vibration peak appears in a wider frequency range, and the peak is obviously enhanced relative to COS, which also shows that the content of N-H in the molecule is increased, thereby indicating that the guanidination reaction occurs on COS amino.
The chemical shift of each C on the chitosan ring is: c1: 97.03, C4: 76.03, C5: 75.01, C3: 71.49, C6: 60.23, C2: 56.23. the chitosan oligosaccharide is obtained by degrading chitosan, so that the chemical environment and chemical shift of each C on a sugar ring of the chitosan oligosaccharide are consistent with those of chitosan. Further determining the structural change of the modified chitosan oligosaccharide by using a 13CNMR analysis technology, wherein the chemical shifts of the chitosan oligosaccharide guanidine salt are respectively as follows: c7: 165.21, C8: 155.13, C1: 98.91, C4: 78.17, C5: 77.03, C3: 69.13, C6: 60.17, C2: 56.43. wherein C7: 165.21 and C8: 155.13 the chemical shift of the newly emerging carbon determines the presence of the guanidine group.
After determining that the chitosan oligosaccharide reacts with dicyandiamide to generate guanidine groups, the invention titrates the product by using NaOH standard solution to measure the guanidine substitution of the productThe degree DS is used as a judgment standard. NH in NaOH and Chitosan oligosaccharide guanidine2 +Cl-Reaction, titration, equivalence point, pH mutation, NaOH mole equal to NH in product2 +Cl-The number of moles. The DS of chitosan oligosaccharide guanidine can be calculated by the following formula:
in the formula:
n1-moles of building blocks substituted with guanidino groups;
n2-moles of building blocks not substituted with guanidino groups;
Δ v — the volume of NaOH consumed to reach the breakthrough point (pH 9.1)/ml;
cNaOHconcentration/mol. L of NaOH Standard solution-1；
G-product mass/G;
318-relative molar Mass of units substituted by guanidino groups/g.mol-1；
161-relative molar masses of structural units which are not substituted by guanidino groups/g.mol-1；
DS- (bis) guanidine degree of substitution/%.
Example 3 the chitosan oligosaccharide biguanide derivative powder COSG prepared in examples 1, 2 was selected for animal pharmacodynamic experiments.
The effect of the present invention will be further described by the following animal experiments.
The experimental animals selected SD adult male rats 70, 10 in each group, 7 in total, cleaning grade, body weight of about 500 gAnimal experiment center in Tianjin. The animals are raised in cages, and 4-5 animals are raised in each cage. The animal feeding laboratory conforms to the national standard clean level and has a space of 30m2The indoor illumination is controlled at the light-dark period rhythm of 12h/12 h. The indoor average temperature is 23 ℃ and the relative humidity is 50-60%.
70 rats were divided into the following 7 groups: control group (labeled C, blank), model group (labeled M, diabetes-inducing and water-filling), metformin group (labeled Met, diabetes-inducing and metformin-administering), chitosan oligosaccharide 1 group (labeled COS1, diabetes-inducing and chitosan oligosaccharide-administering, 1500Da), chitosan oligosaccharide 2 group (labeled COS2, diabetes-inducing and chitosan oligosaccharide-administering, 3000Da), chitosan oligosaccharide guanidine 1 group (labeled COSG1, diabetes-inducing and chitosan oligosaccharide biguanide derivative COSG prepared in example 1), chitosan oligosaccharide guanidine 2 group (labeled COSG2, diabetes-inducing and chitosan oligosaccharide biguanide derivative COSG prepared in example 2). The control group was fed a normal diet and the other groups were fed a high fat and high sugar diet for 8 weeks, followed by fasting for 12 hours, followed by injection of STZ at a dose of 25mg/kg into the tail vein, to construct a type 2 diabetic rat model. When the random blood sugar is larger than 16.7mmol/L, the molding is considered to be successful. In the next 8 weeks, the normal control group was fed with basal diet, the drug-dry group was added with the corresponding drug on the basis of high-fat high-sugar diet, and the drug was gavaged at a dose of 500mg/kg (solvent: distilled water, pH 7), the blank group was gavaged with the corresponding dose of distilled water, and each group of rats was gavaged once a day in the afternoon at the same time.
2. Specimen collection
According to the indexes to be detected in animal experiments, the serum and the tissues of experimental rats are treated as follows for the following detection.
1) Collecting a blood sample: at the end of the experimental period, the rats were fasted for 12 hours and then sacrificed by bleeding under ether anesthesia by femoral artery puncture and blood samples were collected. Standing for 1h, centrifuging at 2500r/min for 20min, and collecting supernatant to obtain serum. Serum samples were stored in a-20 ℃ refrigerator for further analysis.
2) Weight: the weight of the rats was measured at the same time chosen weekly in units: g.
3) liver specimen: after the experiment was completed, rats in each group were exsanguinated and sacrificed, and livers were immediately removed for further analysis.
3. Study conditions
1) Effects on the general condition of diabetic rats:
diabetic rats develop typical symptoms of weight loss. All diabetic rats had significant weight loss (P <0.001) compared to normal rats. The body weight of all other groups of mice gradually went normal during the course of the experiment as compared to the diabetic rats, as shown in figure 3 a. Diabetic rats developed hyperglycemia within 8 weeks and the abdominal blood glucose levels were significantly elevated at the end of the experiment. As can be seen in FIG. 3b, fasting blood glucose levels were significantly increased in group M and decreased after treatment with metformin, COS and COSG (P <0.01) compared to group C. Among them, the COSG2 group with 3000Da performed better in lowering fasting glucose. The effect of metformin, COS and COSG on urine glucose levels is shown in figure 3 c. Urine glucose content in group M was about 1.25 times that in group C (P <0.001), and the drug group exhibited significantly reduced urine glucose levels in diabetic rats (P <0.01) compared to group M. Additionally, the COSG1 group performed best.
To further investigate the effect of chitosan and its derivatives on insulin resistance, insulin and HOMA-IR levels were measured after the end of the experiment. FIG. 4 shows that the insulin content in group M is significantly lower than in group C (p < 0.01). The mean insulin levels in the Met group rats were 26.25% higher than in the M group (p < 0.01). In addition, insulin levels were 30.04% higher in the COSG2 group than in the M group (p < 0.01). The HOMA-IR was significantly increased in the M group rats compared to the C group. However, after 8 weeks of drug treatment, the HOMA-IR decreased to varying degrees, with a COSG of 3000Da having the best therapeutic effect.
2) Influence on IRS-2 expression and Akt phosphorylation in liver of diabetic rat
IRS-2 is a key signaling protein for hepatic insulin resistance, and FIGS. 5a and 5C show the IRS-2 protein expression levels in group C, group M and drug treatment groups. The expression of IRS-2 protein in M group is reduced by 56.26% (p <0.001) compared with C group, but the expression level of IRS-2 protein is increased after treatment. IRS-2 phosphorylation further activates the PI3K/Akt signal transduction pathway. Fig. 5b and 5d show that there was little difference in Akt expression levels between the groups. FIGS. 5b and 5f show that P-Akt (Ser473) protein expression levels were significantly lower than group C (P < 0.01). After Met, COS and COSG treatment, Akt phosphorylation degrees are increased in different degrees; among the treatments, 3000Da COSG had the best therapeutic effect.
3) Influence on the expression level of GLUT-2 and GCK in the liver of diabetic rat
Since insulin resistance is characterized by impaired glucose uptake and since GLUT-2 and GCK play a major role in maintaining hepatic glucose homeostasis, we next examined the effect of COSG on hepatic GLUT-2 and GCK expression levels. As shown in FIGS. 6a and 6b, GLUT-2 expression was significantly reduced in the M group compared to the C group (p < 0.01). Expression of GLUT-2 was up-regulated in each group following metformin, COS, and COSG treatment. Both molecular weight COSG can increase the expression of T2DM rat GLUT-2 protein, but 1500Da COSG has slightly better effect than 3000Da and better effect than Met.
As shown in fig. 6c, GCK expression in the liver of the M group rats showed a somewhat decreased expression (p < 0.01). After the treatment of metformin, COS and COSG, the tendency of the reduction of GCK expression of each group is inhibited, and the intervention effect of the COSG is superior to that of other treatment methods. The therapeutic effects of 1500Da and 3000Da COSG were similar.
4) Influence on the expression level of PEPCK, G6Pase and hepatic glycogen content of diabetic rat liver
Insulin is also known to maintain glucose homeostasis by inhibiting gluconeogenesis, the major target organ for gluconeogenesis. Gluconeogenesis is an anabolic pathway for the formation of glucose, a non-hexose precursor, and is an important cause of glucose production and also an important mechanism for maintaining circulating blood glucose levels. PEPCK and G6Pase are important gluconeogenic enzymes, and their overexpression may lead to the known impairment of glycogen synthesis that affects systemic insulin responsiveness.
As shown in fig. 7a, G6Pase levels were increased in the M group compared to the C group (p < 0.001). The drug-treated group had significantly reduced G6Pase levels compared to group M; in treatment, COSG2 was most effective, slightly better than metformin (p < 0.05). As shown in fig. 7b, PEPCK levels increased by 57.67% (p <0.01) in group M compared to group C. Over-expression of PEPCK was inhibited in the drug-treated group compared to the M group. For liver glycogen content (fig. 7C), liver glycogen content was reduced 33.64% (p <0.01) in group M compared to group C. However, liver glycogen content was significantly increased in the metformin, COS and COSG treated groups compared to the M group. In conclusion, COSG significantly reduced PEPCK and G6Pase levels and increased liver glycogen content. These results indicate that COSG has the ability to reduce hepatic gluconeogenesis, which may further contribute to improving hepatic insulin resistance.
Through the above experiments and analysis, it can be seen that the chitosan oligosaccharide biguanide derivatives of the present invention can control weight loss and reduce fasting blood glucose and urine glucose levels. In addition, it can also increase fasting insulin levels to improve insulin resistance and increase the sensitivity of peripheral tissues to insulin. The research also shows that the COSG can promote phosphorylation activation of position 473 of IRS-2 tryptophan, further activate a PI3K-Akt insulin signal conduction path, on one hand, the expression of downstream signal protein GLUT-2 is improved, and the glucose uptake of the liver is increased; on the other hand, the expression of PEPCK and G6Pase is inhibited, and the hepatic gluconeogenesis is further inhibited. Finally, the chitosan oligosaccharide biguanide derivative has better treatment effect on liver insulin resistance compared with the single use of the traditional diabetes treatment medicament metformin. Namely the application of the chitosan oligosaccharide biguanide derivative in preparing the medicine for treating liver insulin resistance.
The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.
Claims (5)
1. The application of the chitosan oligosaccharide biguanide derivative in preparing the medicine for treating liver insulin resistance is characterized in that the chitosan oligosaccharide biguanide derivative is prepared by taking chitosan oligosaccharide and dicyandiamide as raw materials and taking chitosan oligosaccharide side group NH2Reacting with dicyandiamide under hydrochloric acid and microwave conditions to form a biguanide group bonded with chitosan oligosaccharide, wherein m and n are respectively B in chitosan oligosaccharideDegree of polymerization of the acylation units and deacetylation units.
2. Use of a chitosan oligosaccharide biguanide derivative according to claim 1 in the manufacture of a medicament for the treatment of hepatic insulin resistance, wherein the chitosan oligosaccharide has a weight average molecular weight of 3000 or less, preferably 1500 to 3000 Da.
3. The use of a chitooligosaccharide biguanide derivative according to claim 1 in the preparation of a medicament for the treatment of hepatic insulin resistance, characterized in that the chitosan oligosaccharide deacetylation degree is above 97%, preferably 97-9%.
4. Use of a chitosan oligosaccharide biguanide derivative according to claim 1 in the manufacture of a medicament for the treatment of hepatic insulin resistance, wherein the chitosan oligosaccharide biguanide derivative has a biguanide substitution of 40 to 60%, preferably 45 to 55%.
5. Use of the chitosan oligosaccharide biguanide derivative according to any one of claims 1 to 4 in the preparation of a medicament for treating hepatic insulin resistance, wherein the chitosan oligosaccharide biguanide derivative promotes phosphorylation activation of IRS-2 tryptophan 473, which in turn activates PI3K-Akt insulin signaling pathway, on the one hand, increases expression of downstream signaling protein GLUT-2 and increases glucose uptake by the liver; on the other hand, the expression of PEPCK and G6Pase is inhibited, and the hepatic gluconeogenesis is further inhibited.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003238602A (en) * | 2002-02-20 | 2003-08-27 | Marino Forum 21 | Material for suppressing elevation of blood-sugar level |
| US20110118200A1 (en) * | 2008-06-17 | 2011-05-19 | Zhejiang University | A pegylated and fatty acid grafted chitosan oligosaccharide, synthesis method and application for drug delivery system |
| CN109535279A (en) * | 2017-09-22 | 2019-03-29 | 天津大学 | Chitosan oligosaccharide Biguanide derivative and its microwave synthesis method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003238602A (en) * | 2002-02-20 | 2003-08-27 | Marino Forum 21 | Material for suppressing elevation of blood-sugar level |
| US20110118200A1 (en) * | 2008-06-17 | 2011-05-19 | Zhejiang University | A pegylated and fatty acid grafted chitosan oligosaccharide, synthesis method and application for drug delivery system |
| CN109535279A (en) * | 2017-09-22 | 2019-03-29 | 天津大学 | Chitosan oligosaccharide Biguanide derivative and its microwave synthesis method |
Non-Patent Citations (2)
| Title |
|---|
| LIWANG: "《Microwave-assisted synthesis of chitooligosaccharide guanidine and its effect on GLUT4-dependent glucose uptake through an Aktactivated protein kinase signaling pathway in L6 skeletal muscle cells》", 《RSC ADVANCES》 * |
| 张圣圣: "微波合成壳寡糖双胍对T2DM大鼠胰岛β细胞的修复作用研究", 《中国优秀博硕士学位论文全文数据库（硕士）医药卫生科技辑》 * |
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