CN104558018B - Ascorbic acid vanadyl complex - Google Patents

Abstract

The invention relates to an ascorbic acid vanadyl complex, which is characterized in that the vanadyl group-containing polyhydroxy compound has a molecular formula of Na[VO(C ₆ H ₇ O ₆ ) ₂ OH] and a molecular weight of 443.1. The vanadium compound of the present invention can significantly reduce blood sugar and insulin levels in type II diabetic mice, improve insulin resistance, increase serum adiponectin content, reduce resistin and leptin content, and regulate cholesterol, triglyceride, low-density lipoprotein, HDL content. In summary, the vanadyl ascorbate complex has a good hypoglycemic effect, and can be used to prepare drugs for preventing and treating diabetes, lipid metabolism disorder caused by diabetes and the like.

Description

A kind of vanadyl ascorbate complex

technical field

The invention belongs to the technical field of nutrition product preparation, and in particular relates to a vanadyl ascorbate complex.

Background technique

Diabetes is a complex metabolic disease affected by many factors, which originates from the interaction of environmental factors and genetic factors. The disease can induce a series of complications, such as obesity, cardiovascular and cerebrovascular diseases, hypertension, hyperlipidemia, kidney failure etc. Diabetes is divided into type Ⅰ and type Ⅱ. Most diabetic patients belong to type Ⅱ diabetes. One of the fundamental causes of diabetes is insulin resistance, which runs through the whole process of type Ⅱ diabetes.

Insulin resistance was first proposed by Yallow et al. in the 1950s. It refers to the decrease in the sensitivity of insulin to target organs, the decrease in the efficiency of glucose uptake and utilization, and the compensatory secretion of excessive insulin by the body to produce hyperinsulinemia. , in order to maintain the stability of blood sugar. Typical effector organs of insulin include liver, muscle and fat. Among them, the liver is the first to develop insulin resistance before other target tissues, which leads to the disorder of related enzyme activities involved in glucose metabolism and affects the body's blood sugar level. Cytokines secreted by adipose tissue are transported with the blood to the pancreas, liver, kidney and other organs, causing inflammation in various tissues, causing insulin resistance in various organs and hyperglycemia in the body.

Vanadium is one of the essential trace elements in the human body. It has the function of insulin simulation, and has important biological effects on the growth and development of organisms, glucose metabolism, fat metabolism, cardiovascular and kidney. Under the premise of complete absorption of vanadium, a trace supplement can produce good health care effects. Clinical research points out [Kurt O, Ozden T, Ozsoy N, Tunali S, Can A, Akev N, Yanardag R, Influence of vanadium supplementation on oxidative stress factors in the muscle of STZ-diabetic rats. Biometals: an international journal on the role of metal ions in biology, biochemistry, and medicine.2011,24:943-949], using 1% vanadyl sulfate (100mg/d) of the animal dose to treat diabetic patients for 60 days, effective hypoglycemic and improved Oxidative stress. Experimental results in mice show that vanadate can mimic most of the metabolic activities of insulin, and at relatively high concentrations, vanadate can promote the transport of hexose in fat and muscle tissue; promote the synthesis of glycogen in the liver and diaphragm , inhibit its glycogenolysis; vanadate can promote glucose oxidation and lipogenesis, and inhibit norepinephrine-mediated lipolysis; moreover, due to the decrease of phosphopyruvate kinase mRNA level and the inhibition of glucose-6-phosphatase The inhibition of liver glycogen synthesis caused by vanadium can also be relieved by preventing the decomposition of glycogen [Sakurai H, Tsuchiya K, Nukatsuka M, Sofue M&Kawada J. Insulin-like effect of vanadyl ion onstreptozotocin-induced diabetic rats. The Journal of Endocrinology. 1990, 126:451-459]. In conclusion, vanadium increases the utilization rate of glucose entering these tissue cells by inhibiting the activity of several key metabolic enzyme systems in liver, muscle and adipose tissue, and also reduces the activity of hormones that act oppositely to insulin. Therefore, it is of great significance to provide a vanadium-containing compound that can be stored stably.

Contents of the invention

The purpose of the present invention is to provide a vanadyl ascorbate complex and a preparation method thereof, which can be used for preventing and treating type Ⅱ diabetes caused by insulin resistance.

The vanadyl ascorbic acid complex of the present invention is a polyhydroxyl compound containing vanadyl group (VO ²⁺ ), and its molecular formula is Na[VO(C ₆ H ₇ O ₆ ) ₂ OH].

The preparation method of ascorbic acid vanadyl complex of the present invention is as follows:

Add L-ascorbic acid to acetic acid-sodium acetate buffer solution (pH=5.6), mix well and then add 0.1M pentavalent vanadium salt to react until the color changes from blue to green; then adjust the pH to neutral and add solvent The solution was precipitated, and the precipitate was washed with ethanol and dried to obtain the ascorbyl vanadyl complex.

The pentavalent vanadium salt refers to sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O), ammonium metavanadate (NH ₄ VO ₃ ), sodium metavanadate (NaVO ₃ ·H ₂ O), potassium metavanadate Any one or more of (KVO ₃ ), sodium pyrovanadate (O ₇ V ₂ ·4Na).

The solvent liquid described in the present invention refers to any one in absolute ethanol, normal hexane, acetone, chloroform, sherwood oil, ether.

The adjustment of the pH to neutral is accomplished by adding 0.1M NaOH solution dropwise.

The application of the vanadyl ascorbate complex of the present invention in the preparation of products for treating diabetes and metabolic disorders caused by diabetes.

The vanadyl ascorbate complexes obtained in the present invention can significantly reduce blood sugar and insulin levels in type II diabetic mice, improve insulin resistance, increase serum adiponectin content, reduce resistin, leptin, and TNF-α content, and have The effect of lowering blood sugar.

The present invention uses L-ascorbic acid as a raw material, uses its unique carboxyl group to coordinate and complex it with pentavalent vanadium ions, and obtains an alcohol that can significantly reduce blood sugar and insulin levels in type II diabetic mice and improve insulin resistance. , Fat-soluble compounds. The product of the invention has a good oral effect, especially combines the advantages of ascorbic acid and vanadyl ions, has a good hypoglycemic effect, can be used to prepare medicines for preventing and treating diseases such as diabetes and lipid metabolism disorder caused by diabetes, and has broad market application prospects .

Description of drawings

Figure 1: UV spectrum of ascorbyl vanadyl complex; Ascorbyl vanadyl complex has a special absorption peak of VO ²⁺ at 782nm.

detailed description

The invention will be further described below in conjunction with implementation cases and experimental results. It should be understood that these experimental cases are only used to illustrate the present invention and do not limit the scope of the present invention. The methods for which specific conditions are not indicated in the following examples are carried out by the conditions described in the conventional methods, or according to the conditions described in the conventional methods, or according to the conditions suggested by the manufacturer. Percentages and parts are by weight unless otherwise indicated. Unless otherwise defined, all professional and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. In addition, any method and material that is equivalent to the content described herein can be applied to the present invention, and the preferred implementation methods described in the text are only for demonstration purposes.

Example 1

Weigh 0.88 g of L-ascorbic acid and dissolve it in 100 mL of acetic acid-sodium acetate (NaAc/HAc) buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add 50 mmol/L sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O); after heating at 40° C. for 20 min, the color of the mixture gradually changed from initial blue to green. The pH was adjusted to neutral by dropwise addition of 0.1 M NaOH solution. Anhydrous ethanol solution was slowly added to obtain a green precipitate. Suction filtration, repeated washing with ethanol, and vacuum drying at 55°C for 3 days to obtain the ascorbyl vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the product yield was 96.5%.

Example 2

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40° C. for 20 min, the pH was adjusted to alkaline (pH=9.5) by dropwise adding 0.1 M NaOH solution. Anhydrous ethanol solution was slowly added to obtain a green precipitate. Suction filtration, repeated washing with absolute ethanol, and vacuum drying at 55°C for 3 days to obtain the ascorbate vanadyl complex product. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 33.6%.

Example 3

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40° C. for 20 min, the pH was adjusted to neutral by adding 0.1 M NaOH solution dropwise. Petroleum ether solution was slowly added to obtain a green precipitate. Suction filtration, repeated washing with petroleum ether, and vacuum drying at 55°C for 3 days to obtain the ascorbyl vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 89.5%.

Example 4

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40°C for 20min, adjust the pH to neutral by adding 0.1M NaOH solution dropwise. Cyclohexane solution was slowly added to give a green precipitate. Suction filtration, repeated washing with cyclohexane, and vacuum drying at 55°C for 3 days to obtain the ascorbyl vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 75.4%.

Example 5

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40°C for 20min, adjust the pH to neutral by adding 0.1M NaOH solution dropwise. Chloroform solution was slowly added to give a green precipitate. Suction filtration, repeated washing with chloroform, and vacuum drying at 55°C for 3 days to obtain the ascorbyl vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 72.3%.

Example 6

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40°C for 20min, adjust the pH to neutral by adding 0.1M NaOH solution dropwise. Diethyl ether solution was slowly added to obtain a green precipitate. Suction filtration, repeated washing with ether, and vacuum drying at 55°C for 3 days to obtain the ascorbyl vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 72.3%.

Example 7

Weigh 0.88g of L-ascorbic acid and dissolve it in acetic acid-sodium acetate buffer solution (pH=5.6), stir in a water bath at 40°C for 10 min, and gradually add the same molar amount of sodium orthovanadate (Na ₃ VO ₄ ·12H ₂ O ). The color of the mixture gradually changed from initially blue to green upon initial reaction. After heating at 40°C for 20min, adjust the pH to neutral by adding 0.1M NaOH solution dropwise. Acetone solution was slowly added to give a green precipitate. Suction filtration, repeated washing with acetone, and vacuum drying at 55°C for 3 days to obtain the ascorbate vanadyl complex product. The molecular formula C ₁₂ H ₁₅ O ₁₄ NaV of the ascorbic acid vanadyl complex was calculated according to the elemental analyzer. 50 mg of the sample was dissolved in 1 mL of methanol solution, and vanadyl sulfate solutions with different VO ²⁺ concentrations were used as a standard curve. The absorbance of 782 VO ²⁺ characteristic peaks was measured by spectrophotometry, and the yield was 71.1%.

Effects of the preparations prepared in Examples 1-7 of the present invention on insulin resistance, glucose metabolism and lipid metabolism in high-fat and high-glucose-induced type 2 diabetic mice. The effects of blood glucose levels, insulin levels, adipocytokines (adiponectin, resistin, leptin), and lipid metabolism factors (triglycerides, cholesterol, low-density lipoprotein, high-density lipoprotein) were evaluated. The results showed significant hypoglycemic effect, improvement of insulin resistance and regulation of lipid metabolism. The experimental results are as follows:

Effects of Ascorbyl Vanadyl Complex on Body Weight and Fat Accumulation

Using high-fat and high-sugar diet to induce insulin resistance mouse model in C57BL/6 mice, by comparing with rosiglitazone and measuring the changes of body weight and fat mass to evaluate the effect of improving diabetes and lipid metabolism. As shown in Table 1, the body fat weight of the mice in the model group was significantly higher than that in the normal control group (P<0.01), indicating that the high-fat and high-sugar diet led to fat accumulation. After ingesting ascorbic acid vanadyl complex, compared with the model control group, the body weight of the mice decreased, and the weights of perirenal, epididymis and subcutaneous fat decreased significantly (P<0.01). The results suggest that ascorbyl vanadyl complex can reduce the accumulation of fat in the body.

Table 1 Effects of vanadyl ascorbate complexes on weight gain and fat in insulin resistant mice induced by high fat and high glucose.

^## P<0.01, compared with the normal group; ^* P<0.05, compared with the model group; ^** P<0.01, compared with the model group.

Table 2 Effect of Ascorbyl Vanadyl Complex on Fasting Blood Glucose and Insulin

Using high-fat and high-sugar diet to induce insulin resistance mouse model in C57BL/6 mice, by comparing with rosiglitazone and measuring blood sugar and insulin content to evaluate the effect of improving insulin resistance. High blood sugar and insulin resistance are hallmarks of type 2 diabetes. In the early stages of type 2 diabetes, higher insulin levels indicate more severe insulin resistance. HOMA-IR is an index used to evaluate the level of insulin resistance of an individual. It can be seen from Table 2 that the high-fat and high-sugar diet significantly increased the serum insulin level of the mice in the model group (P<0.01), indicating that the model control group had severe symptoms of diabetes, suggesting that the type 2 diabetes model was successfully established. Compared with the model control group, the serum insulin content and fasting blood glucose of the mice in the ascorbate vanadyl complex group decreased significantly. The results showed that ascorbate vanadyl complex can lower blood sugar and improve insulin resistance in type 2 diabetic mice.

^## P<0.01, compared with the normal group; ^** P<0.01, compared with the model group.

Table 3 Effect of Ascorbyl Vanadyl Complex on Leptin

Leptin is mainly a specific protein secreted by white adipose tissue, which can inhibit the synthesis of fat and is positively correlated with the degree of insulin resistance. It can be seen from Table 3 that after the intervention of the vanadyl ascorbate complex, the leptin content was significantly lower compared with the model group, suggesting that the vanadyl ascorbate complex can inhibit the synthesis of leptin by adipocytes and inhibit insulin resistance.

^## P<0.01, compared with the normal group; ^** P<0.01, compared with the model group.

Table 4 The effect of ascorbate vanadyl complex on adiponectin

Adiponectin is a protein secreted specifically by adipocytes. Serum adiponectin concentration is positively correlated with insulin sensitivity and negatively correlated with the degree of insulin resistance in the body. It can be seen from Table 4 that, compared with the normal group, the serum adiponectin level of the mice in the model group was significantly lower (P<0.01). After the intervention of vanadyl ascorbate complexes, compared with the model group, the level of adiponectin increased, suggesting that vanadyl ascorbate complexes can promote the secretion of adiponectin from adipose tissue and improve insulin resistance.

^## P<0.01, compared with the normal group; ^** P<0.01, compared with the model group.

Table 5 Effect of ascorbyl vanadyl complexes on resistin

Resistin is a specific peptide hormone produced and secreted by adipocytes. It acts on skeletal muscle cells, adipocytes, etc., and can reduce their sensitivity to insulin and improve insulin resistance. It can be seen from Table 5 that after the intervention of the vanadyl ascorbate complex, the resistin content of the mice was significantly lower than that of the model group, indicating that the vanadyl ascorbate complex can reduce the ability of adipocytes to secrete resistin and improve insulin resistance.

^## P<0.01, compared with the normal group; ^** P<0.01, compared with the model group.

Table 6 Effect of Ascorbyl Vanadyl Complex on Liver Total Cholesterol and Triglyceride

It can be seen from Table 6 that after the intervention of the vanadyl ascorbate complex, the total cholesterol content and triglyceride content of the mice were significantly reduced compared with the model group, indicating that the vanadyl ascorbate complex can reduce blood lipids and regulate lipid metabolism.

^## P<0.01, compared with the normal group; ^* P<0.05, compared with the model group; ^** P<0.01, compared with the model group.

Table 7 Effect of Ascorbyl Vanadyl Complex on Liver Low Density Lipoprotein and High Density Lipoprotein

It can be seen from Table 7 that after the intervention of vanadyl ascorbate complexes, compared with the model group, the low-density lipoprotein content of mice was significantly reduced, and the high-density lipoprotein content was significantly increased, indicating that ascorbate vanadyl complexes can regulate cholesterol synthesis and improve the body's Lipid metabolism ability.

^## P<0.01, compared with the normal group; ^* P<0.05, compared with the model group; ^** P<0.01, compared with the model group.

After long-term research by the inventor, it was revealed for the first time that the vanadyl ascorbate complex has remarkable effects on improving insulin resistance, lowering blood sugar and regulating lipid metabolism.

The medicament may be presented in unit dosage form, each containing a predetermined amount of each active ingredient. Such a unit may be adapted to provide 0.5-10 mg/day of the compound, preferably 0.5-1.5 mg/day, 1.0-3.0 mg/day, 2.5-5.0 mg/day, 4.0-8.0 mg/day or 6.0-10.0 mg/day sky. The final dosage will depend on the condition being treated, the route of administration and the age, weight and condition of the patient, and will be at the discretion of the physician. The compounds described herein are most preferably administered in the form of suitable compositions. As suitable compositions there may be mentioned all compositions customary for systemic or local administration. A pharmaceutically acceptable carrier should be substantially inert so as not to interact with the active ingredient. Suitable inert carriers include water, alcohols, polyethylene glycols, gels, and the like. The pharmaceutical preparations may be formulated for administration in any suitable manner, for use in human or veterinary medicine. The pharmaceutical compositions of the present invention may be specifically formulated to be administered in solid or liquid form, including those in the following manner: (1) oral administration, such as drenches (aqueous or non-aqueous solutions or suspensions), tablets , bolus, powder, granule; (2) parenteral administration, such as subcutaneous, intramuscular or intravenous injection, such as sterile solution or suspension. However, in certain embodiments, the host ingredient may simply be dissolved or suspended in sterile water.

The product of the invention is low in production cost and high in safety, can play a good role in preventing and treating insulin resistance and improving glucose and lipid metabolism disorders, and provides a new approach for the development of this type of medicine.

Claims (3)

1. a kind of preparation method of ascorbic acid vanadyl coordination compound, the molecular formula of described ascorbic acid vanadyl coordination compound is na [vo(c₆h₇o₆)₂oh]；It is characterized in that, described preparation method is as follows:

Add l- ascorbic acid in Acetic acid-sodium acetate buffer, after mix homogeneously, add 0.1m pentavalent vanadic salts to be reacted, extremely Color becomes green by blueness；Again ph is adjusted to neutrality, adds solvent liquid to be precipitated, after precipitation washing with alcohol, do Dry obtain ascorbic acid vanadyl coordination compound；Described solvent liquid is dehydrated alcohol, normal hexane, acetone, chloroform, petroleum ether, ether In any one.

2. the method for claim 1 is it is characterised in that described pentavalent vanadic salts is sodium vanadate, ammonium metavanadate, inclined vanadium In sour sodium, potassium metavanadate, sodium pyrovanadate any one or several.

3. the method for claim 1, it is characterised in that described is adjusted to neutrality by ph, is by Deca 0.1m Naoh solution completes.