CN107266598B - Functional sugar prepared based on nanotechnology and application thereof in medical field - Google Patents
Functional sugar prepared based on nanotechnology and application thereof in medical field Download PDFInfo
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- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0087—Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof
- C08B37/009—Konjac gum or konjac mannan, i.e. beta-D-glucose and beta-D-mannose units linked by 1,4 bonds, e.g. from Amorphophallus species; Derivatives thereof
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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Abstract
The invention discloses a method for producing nanometer functional sugar, which comprises a plurality of steps, wherein the raw materials are crushed and processed in a crushing barrel for 60-180 minutes, the rotating speed of a stirring rod is controlled to 2000-6000 rpm, the frequency of ultrasonic waves is controlled to be about 30k-90kHz, and finally the raw materials are refined in a jet mill for 30-120 minutes; finally preparing the nanometer functional sugar with the particle size of 20-40nm, and controlling the molecular weight distribution of the nanometer functional sugar within 10,000-20,000 Da. The nano functional sugar is applied to the product for reducing blood sugar, can obviously reduce blood sugar after being eaten, and has the effect of reducing blood sugar which is within the dosage range of 0.5g/kg to 2.0g/kg and is improved along with the dosage; and carrying out sulfation modification on the nanometer functional sugar to obtain a sulfation derivative with good water solubility, wherein the sulfation derivative has anticoagulation and antithrombotic effects.
Description
Technical Field
The invention relates to a nanometer refining technology, and also relates to functional sugar prepared by the technology and application thereof in medicines.
Background
Nanometer (English: nanometre) is a unit of length, and the international unit system notation is nm. Originally called nano, i.e. 10-9Meters (10 billionth of a meter), i.e. 10-6Mm (1000000 mm). As with centimeters, decimeters, and meters, are measures of length. Corresponding to 4 atomic times in size, and smaller than the length of a single bacterium. The international common name is nanometer, abbreviated nm.
The present scientific research shows that the particle size of the granule can influence the distribution of the medicine in vivo, the particles with the particle size less than 5 μm can pass through the lung, the particles with the particle size less than 300nm can enter the blood circulation, the particles with the particle size less than 100nm can enter the bone marrow, and the nano-medicine can more easily pass through the gastric mucosa, the intestinal mucosa and the nasal mucosa, so that the bioavailability of the oral, nasal administration and transdermal absorption medicine is improved. The nanocrystallization of the particles can present a plurality of excellent properties, specifically represented by quantum size effect, small size effect, macroscopic quantum tunneling effect, surface effect and the like.
The current methods for preparing nanoparticles generally fall into two broad categories: physical methods and chemical methods. The physical method is also called as a crushing method, and is characterized in that solid materials are crushed from big to small to prepare nano powder particles; the chemical method is also called a construction method, and synthesizes the nanometer material by two stages of nucleation and growth of lower limit atoms, ions and molecules. A chemical-based method for producing nano-powder can produce powder of several nanometers. However, the production cost is sometimes high, and the scale-up is difficult, and the particle size distribution is also relatively uneven.
At present, the nano-crushing equipment mainly comprises a multi-dimensional swing type high-energy nano ball mill, a multi-level grading nano ball mill, a high-speed nano crusher, a high-speed shearing superfine crusher, an air flow crusher, an ultrasonic nano crusher and the like.
The main component of the refined powder after the konjak processing is konjak polysaccharide, also called Konjak Glucomannan (KGM), the konjak polysaccharide is natural high molecular polysaccharide with the highest viscosity in known vegetable gum, and is heteropolysaccharide formed by polymerization of glucose and mannose, the average molecular weight is 20-200 ten thousand, the appearance is white or cream to light brown yellow powder, and is formed by polymerization of β - (1, 4) -glycosidic bond of glucose and mannose residue with the molecular ratio of 1: 1.6, branched chains consisting of β (1, 3) glycosidic bond exist on some saccharide residue C-3, 1 branched chain exists on every 32-80 saccharide residues on the main chain, each branched chain has several to dozens of saccharide residues, and about every 19 saccharide residues on the main chain have acetyl group combined by ester bond.
The konjac glucomannan has various excellent characteristics such as gel property, edibility, film forming property and the like, so the konjac glucomannan has wide application in various production fields such as food, medicine, chemical industry and the like. However, KGM has characteristics such as low solubility and poor fluidity, and its application is limited to some extent, and in order to further improve the performance of KGM and expand its application range, it is generally degraded by means such as a physical method, a chemical method, and a biological method.
The existing nano-crushing technology is various, but the molecular weight of nano-particles after crushing cannot be accurately controlled, and measurement and screening are carried out after crushing is needed, so that a needed product is obtained. This consumes a lot of work. At present, due to unhealthy eating habits and living habits, most of human beings have higher and higher blood sugar and need to be treated by medicaments, but the medicaments are basically adverse reactions and are expensive.
In recent years, functional polysaccharides have been actively studied at home and abroad, and many developed countries have defined various polysaccharide products as specific health foods for preventing and treating obesity, hyperglycemia, hyperlipidemia, arteriosclerosis, coronary heart disease and other diseases. The dietary fiber has important significance for human health, and has physiological functions of improving the sensitivity of peripheral nerves to insulin, regulating the blood sugar level of diabetics, increasing the number and phagocytosis of macrophages, improving the disease resistance of human bodies and the like. The konjac mannan is an excellent water-soluble dietary fiber, is suitable for healthy people and diabetic patients to take for a long time, and can be developed and utilized as a health food and a medicine for preventing and treating diabetes.
Because acetyl and a large number of hydroxyl exist in the molecule of the konjac mannan, a series of chemical modifications can be carried out to prepare various derivatives, thereby greatly enriching the research and development and application of the mannan. The original characteristics of the mannan are improved through chemical modification, so that the mannan has better action effect, more importantly, a new derivative can be prepared, and a new attractive product is developed.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for producing nano functional sugar, which can accurately control the molecular weight of a product after being crushed and refined by controlling the particle size of the refined product, so as to obtain the nano functional sugar with concentrated molecular weight distribution; and the device and the method are used for producing nano functional sugar particles in a specified molecular weight range, and after the nano functional sugar particles are taken according to a specified dose, the blood sugar reducing effect is obvious.
In order to solve the technical problem, the invention is solved by the following technical scheme: a method for producing nanometer functional sugar comprises the steps of crushing konjac polysaccharide material in a crushing cylinder for 60-180 minutes, controlling the rotation speed of a stirring rod at 2000-6000 rpm, and controlling the frequency of ultrasonic wave at 30k-90 kHz; secondly, starting a discharging conveyor belt, and conveying the materials on the rotating belt of the stirring blades into a collecting hopper; thirdly, starting a pump to convey the materials in the aggregate bin to a jet mill through a first discharge pipe and to a first high-pressure pump through a second discharge pipe; step four, starting the first high-pressure pump, spraying the material into the jet mill through the first jet pipe to collide with the material output from the first discharge pipe, and then stopping the first high-pressure pump; step five, starting a second high-pressure pump, spraying the materials into the jet mill through a second jet pipe to collide with the materials in the jet mill, and then stopping the second high-pressure pump; step six, starting a third high-pressure pump, spraying the material into the jet mill through a third jet pipe to collide with the material inside, and then stopping the third high-pressure pump; step seven, repeating the steps three to six for 30-120 minutes; and step eight, the material transfer part transports the material collected from the discharge pipe to a finished product bin, and the material in the finished product bin is the nano functional sugar.
The nanometer functional sugar produced by the method has the particle size of 20-40nm and the molecular weight distribution range of 10,000-20,000Da (Dalton). Mannans in this particle size and molecular weight range are referred to as mannosidases or nanomannons.
The method for reducing the blood sugar by using the nano functional sugar has the advantages that the nano functional sugar with the molecular weight distribution of 10,000-20,000Da is applied to a blood sugar reducing product, the blood sugar can be obviously reduced after the nano functional sugar is eaten, and the blood sugar reducing effect is improved along with the dosage within the dosage range of 0.5g/kg to 2.0 g/kg.
The application of the nanometer functional sugar in preparing heparin sodium and heparinoid is characterized in that the nanometer functional sugar with the molecular weight distribution of 10,000-20,000Da is subjected to sulfation modification to obtain a sulfation derivative with good water solubility, and the sulfation derivative has anticoagulant and antithrombotic effects. Because KGM has a skeleton structure similar to heparin and-OH on C2, C3 and C6 in the molecular structure has stronger reaction activity, nano functional sugar with molecular weight distribution of 10,000-20,000Da is subjected to hydroxypropylation and sulfation modification in sequence to obtain KGM propyl aldehyde sulfate sodium salt (heparin sodium) with good water solubility, has anticoagulation and antithrombotic effects similar to heparin, and is an important raw material for preparing heparin sodium and heparinoid.
The invention utilizes a nanometer crushing device to crush the konjac polysaccharide into nanometer particles, controls the particle size of the particles, thereby obtaining the nanometer functional sugar with the molecular weight distribution within 10,000-20,000Da, and has obvious hypoglycemic effect after being taken according to the specified dose; in addition, after derivatization treatment, the derivative has anticoagulation and antithrombotic effects, and is an important raw material for preparing heparin sodium and heparinoid.
Drawings
FIG. 1 is a schematic view showing the structure of a milling cylinder of a nano-milling apparatus used in the present invention.
FIG. 2 is a schematic view showing the structure of a jet mill of the nano-size reduction apparatus used in the present invention.
FIG. 3 is a schematic view of the jet mill of the nano-size reduction apparatus used in the present invention.
Detailed Description
The invention is described in further detail below with reference to the following detailed description and accompanying drawings: as shown in figures 1 to 3, a method for producing nano functional sugar comprises a first step of crushing konjac polysaccharide material in a crushing cylinder 1 for 60-180 minutes, wherein the rotating speed of a stirring rod 13 is 2000-6000 rpm, and the frequency of ultrasonic wave is controlled at 30k-90 kHz; step two, starting the discharging conveyor belt 15, and conveying the materials conveyed by the rotating stirring blades 131 into the aggregate bin 31; step three, starting the pump 3 to convey the material in the collecting hopper 31 to the jet mill 4 through the first discharge pipe 321, and to convey the material to the first high-pressure pump through the second discharge pipe 322; step four, starting the first high-pressure pump, spraying the material into the jet mill 4 through the first gas injection pipe 5 to collide with the material output from the first material discharge pipe 321, and then stopping the first high-pressure pump; step five, starting a second high-pressure pump, spraying the materials into the jet mill 4 through a second gas injection pipe 6 to collide with the materials in the jet mill, and then stopping the second high-pressure pump; step six, starting the third high-pressure pump, spraying the materials into the jet mill 4 through the third air injection pipe 42 to collide with the materials in the jet mill, and then stopping the third high-pressure pump; step seven, repeating the steps three to six for 30-120 minutes; and step eight, the material transfer part transports the material collected from the discharge pipe 41 to a finished product bin, and the material in the finished product bin is the nano functional sugar.
When the invention is used for nano grinding, a nano preparation device is used, which comprises a crushing barrel 1, a stirring rod 13 is arranged in the crushing barrel 1, an upper spiral stirring blade 131 and a lower spiral stirring blade 131 are arranged on the stirring rod 13, a motor 11 is arranged at the bottom of the crushing barrel 1, an output shaft 12 of the motor 11 is connected with the stirring rod 13, a feed pipe 14 is arranged at the bottom of the crushing barrel 1, a discharge conveyor belt 15 is arranged at the top of the crushing barrel 1, the discharge conveyor belt 15 is close to the uppermost end of the stirring blade 131, three circles of ultrasonic parts are annularly arranged on the outer wall of the crushing barrel 1, 7 ultrasonic generators 23 are arranged at each circle of ultrasonic parts, a cooling barrel 2 is arranged outside the crushing barrel 1, a water inlet pipe 21 is arranged at the bottom of the cooling barrel 2, a water outlet pipe 22 is connected on the upper side wall, the other end of the discharge conveyor belt, the discharge part 32 is provided with a first discharge pipe 321 and a second discharge pipe 322, the first discharge pipe 321 is connected with a jet mill 4, the side wall of the jet mill 4 is provided with a first jet pipe 5 and a second jet pipe 6, the top of the jet mill 4 is provided with a third jet pipe 42, the central axes of the first jet pipe 5, the second jet pipe 6 and the third jet pipe 42 are intersected at one point, the second discharge pipe 322 is connected with a first high-pressure pump and a second high-pressure pump, the output end of the first high-pressure pump is connected with the first jet pipe 5, the output end of the second high-pressure pump is connected with the second jet pipe 6, the third jet pipe 42 is connected with a third high-pressure pump, the bottom of the jet mill 4 is provided with a discharge pipe 41, the discharge pipe 41 is connected with a material transfer part, the material transfer part is connected with the first high-pressure pump, the second high-pressure pump and the third high-pressure pump, the material transfer part is also connected with a, a drainage inclined plane 44 is arranged on the flow blocking part 45, an arc-shaped drainage cambered surface 43 is arranged between the drainage inclined plane 44 and the top of the airflow crusher 4, the end part of the drainage cambered surface 43 is connected with a third air jet pipe 42, and grinding balls are arranged in the crushing barrel 1.
The nanometer functional sugar produced by the method has the particle size of 20-40nm and the molecular weight distribution range of 10,000-20,000 Da. Mannans in this particle size and molecular weight range are referred to as mannosidases or nanomannons.
The method for reducing the blood sugar by using the nano functional sugar has the advantages that the nano functional sugar with the molecular weight distribution of 10,000-20,000Da is applied to a blood sugar reducing product, the blood sugar can be obviously reduced after the nano functional sugar is eaten, and the blood sugar reducing effect is improved along with the dosage within the dosage range of 0.5g/kg to 2.0 g/kg.
The application of the nanometer functional sugar in preparing heparin sodium and heparinoid is characterized in that the nanometer functional sugar with the molecular weight distribution of 10,000-20,000Da is subjected to sulfation modification to obtain a sulfation derivative with good water solubility, and the sulfation derivative has anticoagulant and antithrombotic effects. Because KGM has a skeleton structure similar to heparin and-OH on C2, C3 and C6 positions in a molecular structure has stronger reaction activity, nano functional sugar with molecular weight distribution of 10,000-20,000Da is subjected to hydroxypropylation and sulfation modification in sequence to obtain KGM propyl aldehyde sulfate sodium salt (heparin sodium) with good water solubility, and the KGM propyl aldehyde sulfate sodium salt has anticoagulation and antithrombotic effects similar to heparin.
In the prior art, the detection of the molecular weight of polysaccharide is a very complicated and tedious process, and particularly, the detection of the molecular weight of nano functional sugar obtained by a chemical or biological mode is a very difficult operation. According to the device provided by the scheme, the overall particle size distribution of the produced nanometer functional sugar products is similar to normal distribution. After multiple times of detection and comparison, the particle size of the nanometer functional sugar produced by the device through the method is in the range of 20-40nm, and meanwhile, the corresponding molecular weight is distributed between 10,000-20,000 Da. The nanometer functional sugar produced by the device has a corresponding relationship between the particle size and the molecular weight. This greatly simplifies the detection process, as long as the particle size of the particles is detected, thereby saving a large amount of working time. That is, the nano-functional sugar produced by the device can ensure the molecular weight distribution between 10,000 and 20,000Da as long as the particle size is controlled within the range of 20-40 nm.
The invention can obtain the konjac polysaccharide particles with the particle size of 20-40nm by adjusting the pulverizing processing time of the konjac polysaccharide material in the pulverizing barrel 1 to be 60-180 minutes, setting the rotating speed of the stirring rod 13 to be 2000-. The mechanical force forces the macroscopic system of KGM to be cut evenly and orderly, so that the original macromolecular chains and hydrogen bonds are broken to obtain the functional sugar with lower molecular weight.
Experimental embodiment of the present invention 1:
the experimental method comprises the following steps: selecting 100 Kunming adult mice (each half of male and female), wherein the average weight is (25 +/-2.5) g, randomly selecting 10 mice as normal control groups according to the principle of each half of male and female, feeding the rest 40 mice with high-fat feed for 2-3 months, introducing the mice with small dose of streptozotocin to induce diabetes, wherein the fasting blood glucose value is more than 15mmol/L into the experiment, and then randomly dividing the mice into a type 2 diabetes model group, a metformin administration group, a mannan low dose group (0.5g/kg), a mannan high dose group (2.0g/kg), a normal control group and a type 2 diabetes model group, and administering equal amount of distilled water. The experimental period is 4 weeks, and every other week, the blood of each group of mice is collected from the fasting orbit of the eye, and the blood sugar content is measured.
Specific test results are shown in the following table.
TABLE 1 Effect of mannan on blood glucose in type 2 diabetic mice
mmol/L
△ P <0.001, compared to blank group,. P <0.05, compared to model group
As can be seen from Table 1, blood glucose levels of mice in the pre-dose model and each dose group were significantly higher than those in the blank group (P < 0.001); the blood sugar of the metformin administration group is obviously reduced, the blood sugar is obviously reduced at the 1 st week of the administration (P <0.05), and the blood sugar is continuously reduced at the 3 rd week later; the time-dependent relationship of the high-dose administration group of mannan is obvious along with the prolonging of the administration time, the reduction within 4 weeks of administration has obvious difference, the blood sugar change of the low-dose administration group of mannan is not obvious within 3 weeks, and the blood sugar reduction at 4 weeks has obvious difference (P < 0.05). The metformin is a clinical hypoglycemic drug and has an obvious hypoglycemic effect, and comparison shows that the mannan has the hypoglycemic effect and the hypoglycemic effect has an obvious time-effect and dose-effect relationship.
Experimental embodiment of the present invention 2:
48 Kunming adult mice (each half of male and female) are selected, the average weight is (25 +/-2.5) g, the mice are randomly divided into 4 groups according to the principle of each half of male and female, and each group comprises 12 mice. And (3) performing intragastric administration according to the volume of 25ml/kg, respectively administering 10mg/kg of normal saline, 10mg/kg of warfarin, 10mg/kg of low dose of mannan derivatives and 1000g/kg of high dose of mannan derivatives, and continuously performing intragastric administration for 30 d. Blood was collected from the posterior canthus ball venous plexus of mice 1h after administration on day 30, and the clotting time of each group of mice was measured according to the capillary method: and (3) taking blood until the height of a blood column reaches 5cm, breaking off a section of capillary tube every 30s, checking whether the blood coagulation filaments exist, and taking the time from the blood taking to the appearance of the blood coagulation filaments as the blood coagulation time.
TABLE 2 Effect of mannosan derivatives on clotting time in mice
| Group of | Dosage (mg/kg) | Number of animals | Clotting time(s) |
| Physiological saline | 0 | 12 | 222.1±72.3 |
| Hualing Fanhua | 10 | 12 | 295.6±80.3* |
| Product Low dose group | 10 | 12 | 258.7±66.1 |
| High dose group of products | 1000 | 12 | 310.5±53.7* |
P <0.05 compared to saline group
As can be seen from table 2, the clotting times of the low and high dose mannan derivatives were prolonged as compared to the saline group, and increased with the increase of the dose; compared with a control group, the blood coagulation time of the high-dose group mouse of the product has obvious difference, and P is less than 0.05; the warfarin is an anticoagulant medicament used in clinic, has the function of preventing and treating thromboembolic diseases, and shows that the mannan derivative has the function of anticoagulation through comparison. And the anticoagulation has obvious relationship of time effect and dose effect.
The scope of the present invention includes, but is not limited to, the above embodiments, and the present invention is defined by the appended claims, and any alterations, modifications, and improvements that may occur to those skilled in the art are all within the scope of the present invention.
Claims (4)
1. A method for producing a nano-functional sugar, comprising: comprises the steps of crushing and processing the konjac polysaccharide material in a crushing cylinder (1) for 60-180 minutes, controlling the rotation speed of a stirring rod (13) at 2000-6000 rpm, and controlling the frequency of ultrasonic waves at 30k-90 kHz; secondly, starting the discharging conveyor belt (15) and conveying the materials conveyed by the rotating stirring blades (131) into the aggregate bin (31); thirdly, starting a pump (3) to convey the materials in the aggregate bin (31) to a jet mill (4) through a first discharge pipe (321) and to a first high-pressure pump and a second high-pressure pump through a second discharge pipe (322); step four, starting the first high-pressure pump, spraying the material into the jet mill (4) through the first gas injection pipe (5) to collide with the material output from the first material discharge pipe (321), and then stopping the first high-pressure pump; step five, starting a second high-pressure pump, spraying the materials into the jet mill (4) through a second air spraying pipe (6) to collide with the materials in the jet mill, and then stopping the second high-pressure pump; step six, starting a third high-pressure pump, spraying the materials into the jet mill (4) through a third air spraying pipe (42) to collide with the materials in the jet mill, and then stopping the third high-pressure pump; step seven, repeating the steps three to six for 30-120 minutes; step eight, material transfer portion will be from the material transportation of discharging pipe (41) collection to finished product storehouse, and the material in the finished product storehouse is nanometer function sugar, pump (3) are provided with discharge portion (32) are provided with first discharging pipe (321) and second discharging pipe (322), and fluid energy mill (4) lateral wall is provided with first jet-propelled pipe (5) and second jet-propelled pipe (6), and fluid energy mill (4) top is provided with third jet-propelled pipe (42), and the axis of first jet-propelled pipe (5), second jet-propelled pipe (6) and third jet-propelled pipe (42) intersects in a bit, and fluid energy mill (4) bottom is provided with discharging pipe (41), and discharging pipe (41) are connected with material transfer portion, and first high-pressure pump, second high-pressure pump and third high-pressure pump are connected to material transfer portion, material transfer portion still is connected with the finished product storehouse.
2. A nano-functional sugar produced by the method of claim 1, characterized in that: the particle size of the nanometer functional sugar is 20-40nm, and the molecular weight distribution range is 10,000-20,000 Da.
3. Use of the nano-functional sugar according to claim 2 for preparing a hypoglycemic product, wherein: the nanometer functional sugar with the molecular weight distribution of 10,000-20,000Da is applied to the blood sugar reducing product, the blood sugar can be reduced after the product is eaten, and the blood sugar reducing effect is improved along with the dosage within the dosage range of 0.5g/kg to 2.0 g/kg.
4. Use of the nano-functional saccharide according to claim 2 for the preparation of anticoagulant and antithrombotic products, characterized in that: and carrying out sulfation modification on the nanometer functional sugar with the molecular weight distribution of 10,000-20,000Da to obtain a sulfation derivative with good water solubility, wherein the sulfation derivative has anticoagulation and antithrombotic effects.
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