122 results about "1-Deoxynojirimycin" patented technology

The present invention provides a method for increasing the activity of a mutant or wild-type α-glucosidase enzyme in vitro and in vivo by contacting the enzyme with a specific pharmacological chaperone which is a derivative of 1-deoxynojirimycin. The invention also provides a method for the treatment of Pompe disease by administration of chaperone small molecule compound which is a derivative of 1-deoxynojirimycin. The 1-deoxynojirimycin derivative is substituted at the N or C1 position. Combination therapy with replacement α-glucosidase gene or enzyme is also provided.

The invention relates to the field of the traditional Chinese medicine, in particular to a method for extracting and separating 1-deoxynojirimycin (DNJ) with the purity higher than 98% from natural products comprising mulberry leaves, mulberry twigs, white mulberry root-bark, mulberry fruit and silkworm litter and also preparing a 1-deoxynojirimycin crude product (with the purity more than 35%), protein feeds, crude flavone and refined polysaccharide (with the purity more than 90%). The method mainly comprises the following steps: extracting a raw material of dry powder by a hydrophilic solvent, soaking by anion exchange resin after acidizing and concentrating extracting liquid, carrying out chromatography by a cation exchange column after concentrating effluent liquid and water washing liquid, extracting by an organic solvent, recrystallizing, carrying out chromatography by a gel column and purifying. The content of the 1-deoxynojirimycin in the obtained product is not less than 98%,and the concrete execution already enters intermediate test.

The present invention relates to a mulberry leaf effective component composition for reducing blood sugar and its preparation method. Said mulberry leaf effective component composition contains 20-80% of mulberry leaf polysaccharide and 80-20% of mulberry leaf alkaloid in which 1-deoxynojirimycin content is greater than 50%. Said preparation method includes the following steps: removing impurity, drying, pulverizing to obtain mulberry leaf powder, using organic solvent to make extraction, filtering, removing residue, using hot water to make extraction, filtering, adding ethyl alcohol into filtrate to make its concentration be 40%-80%, filtering to obtain supernatant fluid and precipitate, using cation exchange resin and anion exchange resin to make separation and remove impurity, concentrating and drying so as to obtain mulberry leaf alkaloid.

The present invention provides a method for increasing the activity of a mutant or wild-type α-glucosidase enzyme in vitro and in vivo by contacting the enzyme with a specific pharmacological chaperone which is a derivative of 1-deoxynojirimycin. The invention also provides a method for the treatment of Pompe disease by administration of chaperone small molecule compound which is a derivative of 1-deoxynojirimycin. The 1-deoxynojirimycin derivative is substituted at the N or C1 position. Combination therapy with replacement α-glucosidase gene or enzyme is also provided.

The invention relates to a method for continuously extracting and separating 1-deoxynojirimycin (DNJ) and flavone from folium mori; the key points are that: folium mori and ethanol are mixed, and extracted and filtered centrifugally, and then flocculating agents are added for flocculation and precipitation after being concentrated; the precipitation is removed, and the filterate is added in a plurality of cation exchange resin columns which are communicated, and flows through a macroporous absorption resin column connected with the resin columns in series; after the filtrate adding is finished, the filtrate is washed until outflow liquid from the macroporous absorption resin column is colourless, and then the macroporous absorption resin column is disconnected, and then the 1-deoxynojirimycin (DNJ) is eluated with ammonia water or ammoniac ethanol solution from the cation exchange resin columns; the eluate is eluated by the cation exchange resin columns again, the operation is repeated, and then the eluate is collected, is concentrated and filtered with a 80D nanofiltration membrane and then is spray-dried, so as to obtain claybank powdery 1-deoxynojirimycin (DNJ). 3-10 percent ofalcohol is used for washing the macroporous absorption resin column to remove colloidal impurities, and then 50-70 percent of alcohol is used for washing flavone, the claybank eluate is collected, decompression and concentration are carried out to recover alcohol, and then spray-drying is carried out to obtain the claybank powdery folium mori general flavone.

The invention discloses anextract of Chinese herbs containing alpha-glycoside enzyme inhibitor, which comprises cytidine, 2-(1',2',3',4'-tetrahydroxylbutyl)-5-(2'',3'',4''-trihydroxybutyl)-pyrazine and 1-deoxynojirimycin. The extract can be obtained from leaves, branches and peels of mulberries. The extract can be used for preparing medicament for treating diabetes.

The invention discloses a method for extracting and separating 1-deoxynojirimycin from mulberry leaves, which comprises the following steps of: pretreating a raw material, namely crushing a mulberry leaf raw material; performing enzymolysis; extracting by using alkali-alcohol, cooling, filtering to obtain extracting solution; concentrating, and performing acid dissolution and ultrafiltration to obtain ultrafiltrate; removing impurities by using cation resin, namely making the obtained ultrafiltrate pass through the cation resin, eluting by using ammonia alcohol solution, collecting eluent; making the eluent pass through macroporous adsorption resin, eluting by using ethanol solution, and collecting eluent; and concentrating the eluent, adding a methanol-saturated ammonia water mixed solvent, regulating temperature, cooling for crystallization to obtain crystals, and drying. A product with the purity of over 95 percent can be extracted and separated from the mulberry leaf raw material,and the 1-deoxynojirimycin is effectively separated; requirements on equipment are low, and the operation is safe; the process is simple, the used solvents can be recycled, and the production cost isreduced; and the used solvents are low in toxicity, environment-friendly and safe.

The invention provides mulberry leaf buccal tablet and a preparation method thereof. The components of the mulberry leaf buccal tablet are as follows: mulberry leaf instant powder, thinner, flavoring agent and forming agent. The preparation method has reasonable design, simple and convenient operation, and quite low production cost. The prepared mulberry leaf buccal tablet contains nutrient, such as mulberry leaf water extracted component flavone, 1-deoxynojirimycin, amino acid, mulberry leaf amylase, etc. with the added auxiliary materials suitable to be eaten by diabetic patients, has the functions of dispelling wind-heat, clearing lung-heat and moistening dryness, clearing liver and brightening eyes, inhibiting higher blood glucose, etc., is soluble right after being taken, is good for keeping oral cavity sanitary, has good taste, is rich in nutrient and has good health care effect.

The invention provides a composition of plant extract and a method for regulating and controlling blood sugar by using the same. The composition of a plant extract comprises a glycosidase inhibitor used for blocking digestion and absorption of sugar by organism, and a sodium glucose transporter inhibitor used for inhibiting sodium glucose transporter so as to inhibit the reabsorption of the glucose by kidney. The glycosidase inhibitor comprises L-arabinose, 1-deoxynojirimycin, phaseolin or chlorogenic acid; the sodium glucose transporter inhibitor is extracted from phlorizin; the phlorizin is extracted from lithocarpus polysachyus rehd, branches or leaves of a fruit tree. On the one hand, the composition provided by the invention can be used for blocking excessive absorption of the sugar, on the other hand, the composition can be used for inhibiting the sodium glucose transporter in the kidney to control the reabsorption of the glucose so as to ultimately lower blood sugar and balance the blood glucose level within the scope of health.

The invention relates to a purification technology for extracting a compound from plants, particularly for extracting and separating high-purity 1-deoxynojirimycin from natural products, namely mulberry leaves, and simultaneously refining mulberry leaf polysaccharide. The method mainly comprises the following steps: pretreating raw materials, then dissolving the raw materials in an ethanol solution, performing ultrasonic extraction, concentrating extract liquid, then enabling the extract liquid to pass through cationic resin, concentrating eluate, and then extracting with an organic solvent, recrystallizing and carrying out gel filtration chromatography to prepare a refined product of the 1-deoxynojirimycin with the purity not lower than 98.5%. The process is simple to operate, low in cost, low in environmental pollution and typical in equipment and the materials are easy to obtain; therefore the process is suitable for large-scale production.

The invention discloses a solid or liquid medicinal composition for non-gastrointestinal administration. The medicinal composition contains 1-deoxynojirimycin component and / or pharmaceutically acceptable salt thereof, and the 1-deoxynojirimycin component and / or the pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient are prepared into the medicinal composition for non-gastrointestinal administration together. The medicinal composition for non-gastrointestinal administration can be injection, pills, sublingual tablets, oral patches, buccal tablets, respiratory inhalation aerosol, powder for inhalation, suppository and capsules for rectums and transdermal preparations. The medicinal composition can be used for preventing and / or treating virus infection of mammals comprising human in a non-gastrointestinal administration mode, and the composition has good broad-spectrum antivirus effect and high safety.

The invention provides a bacillus amyloliquefaciens engineering strain for producing 1-deoxynojirimycin at high yield and a fermentation method. The preservation number is CCTCC NO.M2017849, and the bacillus amyloliquefaciens engineering strain is named bacillus amyloliquefaciens HZ-15, and the invention further provides a construction method of bacillus amyloliquefaciens HZ-15 and a method for producing 1-deoxynojirimycin (DNJ) at high yield. By the adoption of a molecular biological technique, on the basis of preserved bacillus amyloliquefaciens LX-12 for producing DNJ in a laboratory, the engineering strain HZ-15 for producing the DNJ at high yield is constructed successfully through a homologous recombination method. With soybeans being raw materials, solid fermentation is performed; by adding 1% of malt extract powder and 2% of lactose into a solid fermentation culture medium, the yield of the DNJ is up to 1117.2 mg / kg.

The invention discloses a method for extracting high purity 1-deoxynojirimycin. The method comprises the following steps: pretreating raw materials; conducting enzymolysis digestion; acidifying a filtrate; filling the filtrate into a strong acidic cationic resin column; eluting with ammonia to obtain an ammonia eluate; conducting membrane separation on the ammonia eluate to remove impurities, and concentrating; conducting column chromatography on the concentrated solution by a weak acidic cationic resin column; eluting by ammonia water; collecting the eluate by segment; and concentrating, crystallizing and drying to obtain a 1-deoxynojirimycin refined product. The enzymolysis digestion step uses a combined enzyme comprising cellulase, glucanase, pectinase in a weight ratio of 3-5:2-4:1. The method for extracting high purity 1-deoxynojirimycin provided by the invention uses an enzymolysis method, which is organically combined with the technologies of impurity removal and purification; therefore, the extraction method provided by the invention has the advantages of high extraction rate and purity higher than 98% of a target product, no environmental pollution and improved product quality and safety.

The invention relates to a method for continuously extracting and separating polysaccharides and 1-deoxynojirimycin from mulberry leaves. The method comprises the following steps: grinding the mulberry leaves, carrying out supercritical CO2 extraction under certain conditions and collecting mulberry leaf dregs; after drying the mulberry leaf dregs, carrying out extraction through ethanol refluxing; concentrating the extracting solution, standing and drying the precipitate, thus obtaining a polysaccharide compound with the content being 60-80%; concentrating the supernatant and adsorbing active ingredients on resins via cation exchange resins; eluting the resins with water-containing ethanol and ammonia water; and concentrating the ammonia water eluent with a reversed-phase C18 silica gel based chromatographic column and drying the concentrated products, thus obtaining alkaloids in the mulberry leaves, wherein the content of 1-deoxynojirimycin in the alkaloids in the mulberry leaves is 80-86%. The detailed method refers to the specification. The method has the advantages that the process is simple, the extraction rate is high, the obtained products have higher purity, and the resins and silica gel used for purification are renewable, low in costs and beneficial to industrial popularization and application.

The invention discloses a response surface method optimized method for extracting 1-deoxynojirimycin (1-DNJ) in mulberry leaves with an acidic ethanol solution. The method is characterized in that the acidic ethanol solution is taken as the extraction solvent for extracting the 1-DNJ in the mulberry leaves, and the extraction condition is optimized by the response surface method so that the optimal condition is obtained, namely, the solution-to-material ratio is 25, and in other words, 25mL of extraction solvent is used for each gram of crude mulberry leaf powder; the method comprises the steps of extracting for 2.7h under the condition of 80 DEG C, re-extracting once, purifying the filtrate, and then concentrating the filtrate to obtain the 1-DNJ extract; the method is capable of obtaining about 0.2% of mulberry leaf 1-DNJ extraction rate, which is remarkably higher than the traditional ethanol extraction rate 0.00139% and the diluted acid extraction efficiency 0.00147%; obviously, the diluted acid and the ethanol have synergistic effect in the mulberry leaf 1-DNH extraction process and unexpected effect is generated, and therefore, theoretical support and technical guidance are provided for development and utilization of the mulberry leaf 1-DNH in the aspects of medicine and healthcare products.

An edible product for normalizing blood sugar levels is provided. The product includes a mulberry extract having a (w / w) flavonoids concentration of at least 10%, a 1-deoxynojirimycin (DNJ) concentration of about 2% and a GABA concentration of about 1% of the total amino acid content of said mulberry extract.

The present invention provides a method for increasing the activity of a mutant or wild-type a-glucosidase enzyme in vitro and in vivo by contacting the enzyme with a specific pharmacological chaperone which is a derivative of 1- deoxynojirimycin. The invention also provides a method for the treatment of Pompe disease by administration of chaperone small molecule compound which is a derivative of 1-deoxynojirimycin. The 1-deoxynojirimycin derivative is substituted at the N or Cl position. Combination therapy with replacement a-glucosidase gene or enzyme is also provided.

The present invention relates to a polypeptide associated with the biosynthesis of 1-deoxynojirimycin which is a substance that inhibits a-glucosidase activity, to a polynucleotide coding therefor, to a vector comprising the polynucleotide, to a transformant comprising the vector, and to a method in which the transformant is used in order to produce either a polypeptide associated with the synthesis of 1-deoxynojirimycin or 1-deoxynojirimycin. The polynucleotide of the present invention can be used in the volume production of 1-deoxynojirimycin, or can be used in studies searching for novel 1-deoxynojirimycin derivatives using recombinant technology, and hence is useful in developing pathogenic virus inhibitors and hypoglycaemic agents for diabetic patients.

The invention relates to a technological method for extracting 1-deoxynojirimycin from mulberry leaves, which is a technological method for extracting and separating 1-deoxynojirimycin with content being more than 70 percent from mulberry leaves. The technological method comprises the following steps of: conducting water extraction and alcohol precipitation on mulberry leaves, removing impurities by using macroporous resin, enabling mulberry leaf alkaloid to be enriched by cations, conducting gradient elution through silica gel column chromatography, and conducting vacuum drying to finally obtain the 1-deoxynojirimycin product. The technological method provided by the invention has the advantages that the technological process is simple, the obtained product does not contain residual organic solvent, the fillers for silica gel column chromatography are low in cost and can be repetitively used, the cost is reduced, special equipment is not needed in the entire technological process, the operation is simple and feasible to conduct and the technological method is very suitable for industrial production.

The invention relates to the field of separation and purification of active ingredients of a natural product, in particular to a method for preparing high-purity 1-deoxynojirimycin with a combined membrane separation and column chromatography technology. According to the method, raw materials of silkworm and white mulberry is subjected to water extraction in aid of ultrasonic waves; an ultrafiltration feeding liquid is obtained through centrifugation and micro-filtration membrane treatment; and a 1-deoxynojirimycin product with purity larger than 80% is obtained through steps of secondary ultrafiltration, nanofiltration, cation exchange resin chromatography and dextrane gel chromatography. The method has the advantages of simple process, high efficiency, safety and environment protection.

The invention relates to a method of cultivation of Cordyceps Militaris of 1-deoxynojirimycin enrichment. The method of cultivation of Cordyceps Militaris containing rich effective active components of Mulberry leaves comprises the steps that 1, raw materials rich in active Mulberry leaves ingredients are added into a culture medium, sterilization is conducted, and enrichment culture medium is prepared; 2, the fruiting body of Cordyceps Militaris is taken, after the cultivation, Cordyceps fungus liquid is prepared; 3, Cordyceps fungus is inoculated into enrichment culture medium, light cultivating is conducted for 3-5 days; culturing is conducted for 2.5-3.5 days; sprouting management is conducted for 14-26 days; harvest is obtained. According to the method of cultivation of Cordyceps Militaris of 1-deoxynojirimycin enrichment, through the addition of Mulberry leaves or Mulberry leaf extract in traditional wheat culture substrate of Cordyceps, cultivation of Cordyceps Militaris is conducted, Cordyceps Militaris of 1-deoxynojirimycin enrichment with active ingredients from mulberry leaves is obtained; after the testing, the contained 1-deoxynojirimycin proportion meets the requirement of traditional Chinese medicine medication, the amount of drug dose is reduced, and the medicinal value of Cordyceps Militaris is dramatically improved.

The invention belongs to the field of detection of quality of traditional Chinese preparations, and particularly relates to a method for detecting a pharmaceutical preparation for treating the xiaoke disease. The pharmaceutical preparation comprises the active pharmaceutical ingredients, by weight, 6000-8000 parts of silkworm feces and 100-200 parts of radix glycyrrhizae. According to the method for detecting the pharmaceutical preparation, the pharmaceutical preparation is detected by measuring the content of 1-Deoxynojirimycin through an HPLC method. The method for detecting the pharmaceutical preparation is rapid, comprehensive and highly targeted, the quality of the preparation can be effectively controlled, and use safety and stability of the preparation can be improved.

The invention discloses a medicine composition and application thereof, and a medicament containing the medicine composition. The medicine composition comprises moraceae morus plant total alkaloids and balsam pear total saponins, wherein the mass content ratioof the moraceae morus plant total alkaloids and the balsam pear total saponins is 1:2-7:1; the moraceae morus plant total alkaloids are used in the form of moraceae morus plant extracts in the medicine composition, and the balsam pear total saponins are used in the form of balsam pear extracts in the medicine composition; the moraceae morus plant total alkaloids comprise 1-deoxynojirimycin, and the mass content of the 1-deoxynojirimycin in the moraceae morus plant extracts is 2-50%. Target components are extracted from natural plants; the medicine composition has excellent effects of decreasing blood-glucose, losing weight, and reducing lipid in a synergetic manner, and is particularly suitable for the treating and preventing type 2 diabetes.

A determination method of the 1-deoxynojirimycin content in silkworm powder relates to silkworm powder. The method comprises the following steps: weighing two 1-deoxynojirimycin control samples used as control liquid 1 and 2; sucking the control liquid 1 and 2, injecting the samples; weighing silkworm powder to be determined, adding petroleum ether, performing heating reflux and extraction in water bath, filtering, removing the filtrate, volatilizing the petroleum ether to dryness to obtain a product A; adding methanol in A, performing ultrasonic extraction, filtering, concentrating the filtrate, adding in a neutral alumina column, performing elution, collecting the eluted solution, evaporate to dryness, dissolving by methanol and obtaining a constant volume so as to used the solution as a test solution; performing HPLC analysis of the test solution, calculating the 1-deoxynojirimycin content in the silkworm powder to be determined by an external standard two-point method according to a peak area value. The method has the advantages of high precision, good repeatability and stability, qualified sample recovery rate, simplicity and practicality, and the like, can be popularized in general laboratories, and is applicable to the quality control of 1-deoxynojirimycin in silkworm powder products.

The invention provides a cultivation method for increasing the yield of vegetable mulberry tender shoot and improving the quality of the vegetable mulberry tender shoot. Compared with the traditionalmulberry cultivation methods, the cultivation method has the advantages that growth habit of mulberries and quality requirements on pollution-free vegetables are combined with one another, the yield of the mulberry tender shoot can be obviously increased by the aid of the cultivation method, and the eating quality of mulberry shoot can be improved by the aid of the cultivation method; special mulberry varieties are selected, mulberry tree form development modes are changed, and special fertilizers are applied; the yield of the mulberry shoot can be obviously increased, taste of the mulberry shoot can be improved, and the content of DNJ (1-deoxynojirimycin) in the mulberry shoot can be increased.

The invention discloses a method used for high throughput screening of bacterial strains capable of generating 1-deoxynojirimycin (DNJ). The method comprises following steps: 1, recombinant vectors used for expressing DNJ biosynthetic pathway and original strains used for expressing beta-galactosidase are constructed; 2) error-prone PCR products of corresponding DNJ synthetic genes are obtained; 3, the error-prone PCR products of corresponding DNJ synthetic genes are taken as primers, the recombinant vectors are taken as templates, and MEGAWHOP PCR is carried out so as to obtain an MEGAWHOP PCR plasmid mutant library; and 4, the MEGAWHOP PCR plasmid mutant library is inserted into the original strains, plate coating is carried out so as to obtain a mutant library plate, bacterial colonies on the mutant library plate with color lighter than that of bacterial colonies on a wild bacterial plate are collected, and the collected bacterial colonies are high DNJ yield bacteria. The method can be used for screening bacteria strains high in DNJ yield, low in DNJ yield, or producing no DNJ; screening efficiency is increased; time is saved; and screening steps are simplified.

The present invention discloses an application of 1-deoxynojirimycin in preparation of medicine for curing diabetes and nephropathy. Said invention also discloses medicine composition for effectively curing diabetes and nephropathy. It contains 1-deoxynojirimycin with safe effective dose and pharmaceutically-acceptable carrier or excipient.

The invention relates to a blood glucose adjusting health food taking mulberry seed powder as an effective component, which comprises the following components in parts by weight: 4-99.9 parts of mulberry seed powder, 0-10 parts of flavor agent and 0.1-96 parts of accessory substance. The mulberry seed contains 1-deoxynojirimycin (DNJ) with high concentration, can inhibit the activity of small intestine alpha-glycosidase and can adjust blood glucose, thereby having obvious effect on treating human diabetes and obesity. In the invention, the mulberry seed powder is combined with the flavor agent, the accessory substance and natural products are processed into different types of health foods with the function of adjusting human blood glucose without any toxic or side effect, the safety is high, the taste flavor is good, the edibility is strong so as to control the blood glucose of a diabetic patient in the normal diet process, and simultaneously, natural resources are developed.

The invention mainly relates to a method for extracting 1-deoxynojirimycin and resveratrol from mulberry leaf, and provides a whole set of method for separating and extracting high-purity 1-deoxynojirimycin and resveratrol from mulberry leaf. By the method, the yield and purity of 1-deoxynojirimycin and resveratrol are greatly increased. The method comprises the following steps of: carrying out alcohol reflux extraction, film filtration, concentration, extraction and activated carbon decolorizing, and then controlling crystallization and recrystallization conditions to separately obtain 1-deoxynojirimycin and resveratrol pure products. The method has the advantages that: in the whole separation process, the requirements on the ambient conditions and equipments are low, the separation timeis shortest, the product purity is highest, and the comprehensive utilization rate of raw materials is highest; and moreover, the separation materials are obtained easily and inexpensive, the separation operation process is simple and easy to control, crystallization and recrystallization are adopted, the purification efficiency is high, used reagents are nontoxic and safe, and the cost is low.