53 results about "Dehydrocholesterols" patented technology

The invention discloses a preparation method of a 7-dehydrocholesterol, which comprises the following steps: a) protecting a hydroxyl on the 3 position of a cholesterol; b) oxidizing a carbon on the 7 position of the product made in step a) into a carbonyl; c) carrying out hydrazone reaction to the product made in step b) with hydrazine derivatives, the structure of the hydrazine derivatives being at most one H to be substituted by the following hydrazine; d) reacting the product made in step c) with a strong basic reagent having stronger alkaline than a sodium hydroxide; e) reacting the product made in step d) with water or an acid. In the preparation method, no halogen group elements such as Br element are participated, the side effects are less and the yield is high, and the prepared product is safe and innocuous.

The present invention relates to the field of pharmaceutical chemistry, and in particular to methods of preparing cholesterol,and derivatives and analogs thereof. The cholesterol derivatives include, but not limited to, 7-dehydrocholesterol, 25-hydroxycholesterol, 25- hydroxy7dehydrocholesterol and ergosterol. In the invention, phytosterol can be used as a raw material to prepare the compound shown in the formula I through microbial conversion, and then cholesterol and the derivatives and analogues thereof are prepared.

An implant to be used as medical or dental implant, comprising a metallic or polymeric base which is covered by the vitamin D precursor cholecalciferol. The implant can be obtained by direct covering of the polymeric or metallic base with a solution comprising cholecalciferol or also covering the base with the 7-dehydrocholesterol (7-DHC), and subsequently irradiated with UV light to induce the formation of cholecalciferol. Optionally, the coating of the implant may include an antioxidant such as vitamin E. This implant enhances osseointegration in compromised patients by means of the endogenous synthesis and activity of vitamin D in hard and mineralized tissue regeneration. Furthermore, a method to obtain these implants which comprises coating the surface of the implant directly with cholecalciferol or with a specific concentration of 7-DHC and irradiated with UV light to induce the formation of cholecalciferol.

The invention relates to a method for increasing the yield of 7-dehydrocholesterol in saccharomycetes by using compartmentalization. The method comprises the following steps of: by taking saccharomycetes as an original strain, expressing heterologous sterol delta 24-reductase and delta-cholestenase, wherein the yield of 7-DHC in saccharomyces cerevisiae S288C is detected to be 10.15mg / L. According to the method disclosed by the invention, partial enzymes in a 7-DHC synthesis path are positioned in each compartmentalization in the saccharomyces cerevisiae by using an oxidase and a mitochondrial positioning tag, and a relatively independent 7-DHC synthesis path is formed. Meanwhile, the storage space of precursor substances needed by 7-DHC synthesis is increased, the feedback effect is reduced, the conversion efficiency between enzymes is improved in the same compartment, the loss of the acting substrate is reduced, and finally the yield of the 7-DHC is improved by 4 times and reaches 53.31mg / L.

According to the present invention, there can be provided A process for producing 7-dehydrocholesterol (hereinafter, "7DHC"), comprising culturing, in a medium, a 7DHC-producing Labyrinthulea microorganism in which the 7DHC reducing activity is reduced or lost as compared to a parent strain through deletion, substitution, or addition of at least one base in a gene which is present in the chromosomal DNA of the parent strain and encodes a protein having 7DHC reducing activity, and the microorganism produces 7DHC, allowing 7DHC to be produced and accumulated in the culture, and collecting the 7DHC from the culture; and a process for producing vitamin D3, comprising irradiating, with ultraviolet light, the 7DHC produced by the production process.

The invention relates to belongs to 24-dehydrocholesterol reductase coded recombinant adenoviruses specifically expressed in tissue, which belongs to the Alzheimer's disease gene treatment method and the treatment field. More specifically, according to the invention, by aid of a gene adenovirus carrier, the coded gene of 24-dehydrocholesterol reductase (DHCR 24) is introduced in nerve cells, the expression of 24-dehydrocholesterol protein in nerve cells is regulated, and the nerve cells protection effect can be performed. The invention also relates to a construction of defective recombinant adenoviruses for replication comprising nerve specifically expressed DHCR 24 gene and an application of the carriers in DHCR 24 gene in cells and an application of the DHCR 24 protein in nerve cells specific expression. The prepared recombinant adenoviruses are expected to obtain positive curative effect in neurodegenerative diseases such as Alzheimer's disease or other gene treatments which require a DHCR 24 supplementary therapy.

The invention relates to the field of microbial metabolism, and particularly relates to saccharomyces cerevisiaes for producing 7-dehydrocholesterol and a construction method thereof. The invention discloses two recombinant saccharomyces cerevisiaes for producing 7-dehydrocholesterol, named as saccharomyces cerevisiae SyBE_Sc01250043 and saccharomyces cerevisiae SyBE_Sc01250045, or SyBE_Sc01250043 and SyBE_Sc01250045 for short, as well as a construction method thereof. Compared with the existing reported strains, the recombinant saccharomyces cerevisiaes of the present invention has the advantage of significantly improving yield of 7-dehydrocholesterol.

The invention relates to a method for virtually screening cholesterol degrading medicine with 24-dehydrocholesterol reductase (DHCR24) being a target point. The method includes the steps that 1, the molecular structure of DHCR24 is determined; 2, molecular docking software is adopted, the active center of macromolecular docking of the medicine is determined according to a combined pocket which is formed after desmosterol and DHCR24 are docked, and an active pocket is set; 3, a macromolecular ligand databse used for docking is arranged; 4, according to the set active pocket, by the utilization of the molecular docking software, macromolecular ligands in the macromolecular ligand database used for docking and the active pocket are docked in sequence; 5, results which are screened after primary docking are accurately docked, and the candidate medicine having the effect of cholesterol degrading are determined. By the adoption of the method, within short time, the macromolecular candidate medicine which has the effect of competitive restraining of DHCR24 and is developed into the novel cholesterol degrading medicine is obtained, in this way, the research and development efficiency is greatly improved, and the research period of new medicine is shortened.

A health-care fluorescent-lamp powder can prevent osteoporosis. It is a kind of alkali boron phosphate with expression formula ((Mo)1-3. (B203)1-4. (P205)0.1-1:Pb0.005-1. Y0.01-1), M belongs to one or above of calcium, magnesium, strontium and sulfone, and Y is one or above of rare-earth ion lanthanum, samarium and gadolinium. The process is carried out by mixing one or above of calcium carbonate, strontium carbonate and barium carbonate with one or above of magnesium oxide, boric acid, ammonium dibasic phosphate, lead oxide, lanthanum oxide, gadolinium oxide and samarium sesquioxide and solid-phase firing at high-temperature. It can transmit medium-wave ultraviolet with peak value=2970, convert human-body hypodermic 7-deoxycholesterol into vitaminD3, and improve human-body calcium-phosphorus metabolism. It can be used for health-care fluorescent-lamp in family and hospital.

The invention belongs to the technical field of biochemical engineering and enzyme engineering, and in particular relates to a method utilizing the microsomal enzyme of an animal liver to biosynthesize 7-dehydrocholesterol. The method includes the following steps: (1) preparation of microsomal enzyme solution; (2) enzymatic conversion reaction; (3) saponification reaction; and (4) extraction. The method can prepare the 7-dehydrocholesterol which is completely identical with that prepared by chemosynthesis; and meanwhile, the technology is simple and easy, and not only the production cost of vitamin D3 can be reduced, but also the environment pollution can be reduced.

The invention relates to a tobacco 7-dehydrocholesterol reductase gene and application thereof. The nucleotide sequence of the 7-dehydrocholesterol reductase gene is as shown in SEQ ID NO. 1. Based on the important influence of sterol on the safety of tobacco leaves, the tobacco sterol metabolism regulation gene is deeply studied, so that a new tobacco variety is constructed by utilizing genetic engineering, and a good application foundation is laid for improving the tobacco variety. According to the application, preliminary research on specific tobacco sterol 7-dehydrocholesterol reductase shows that the specific tobacco sterol 7-dehydrocholesterol reductase is highly related to the sterol content in tobacco leaves, when the gene is silenced in Bensi tobacco, the stigmasterol content and the campesterol content in the tobacco leaves are reduced, and the total sterol content is reduced by 24.8%. Based on the characteristic, a certain foundation and reference can be provided for tobacco quality regulation and control and new tobacco variety cultivation.

The invention relates to an isolated, genetically modified, living non-mammal organism, having increased HMG-CoA-reductase activity compared to the wild type, and having reduced C24-methyltransferase and / or delta22-desaturase activity compared to the wild type. The invention is characterized in that the organism has increased dehydrocholesterol-delta70-reductase activity compared to the wild type. The invention further relates to different uses of such an organism, to a test kit comprising such an organism, and to a membrane extract of such an organism.

The invention relates to the field of organic synthesis, and discloses a method for preparing acetyl cholesterol-7-one p-toluenesulfonylhydrazone. The method comprises the following steps: carrying out a contact reaction on acetyl cholesterol-7-one and p-methylbenzenesulfonhydrazide under a condensation reaction condition in a solvent containing cation exchange resin. According to the method for preparing the acetyl cholesterol-7-one p-toluenesulfonylhydrazone, acetyl cholesterol-7-one p-toluenesulfonylhydrazone with a relatively high content can be obtained under mild reaction conditions, sothat the yield of subsequent preparation of 7-dehydrocholesterol is improved. Meanwhile, a catalyst in the method is easy to recycle after the reaction, so that overall cost is reduced.

The invention discloses a liquid 25-hydroxyvitamin D calibrator diluent, which comprises the following components: a buffer solution, a protein stabilizer, a preservative and a cyclopentane polyhydrophenanthrene stabilizer. The main innovation of the invention lies in that a) 7-dehydrocholesterol is applied to the 25-hydroxyvitamin D calibrator for the first time for improving the stability of 25-hydroxyvitamin D under a liquid aerobic condition; b) all the components in the calibrator diluent are optimal formulas, so that the stability of the 25-hydroxyvitamin D calibrator is powerfully guaranteed, and the 25-hydroxyvitamin D calibrator can be stably stored at 8 DEG C for 12 months; and c) the liquid 25-hydroxyvitamin D calibrator is applied to detection of 25-hydroxy vitamin D in the serum sample, the calibrator does not need to be redissolved during detection, and the repeatability of a calibration curve which is calibrated for multiple times is superior to that of the calibration curve of the freeze-dried calibrator.

The present invention discloses a method for synthesizing 25-hydroxy cholesterol. The method is as below: subjecting 24-dehydrocholesterol derivative as a raw material, which undergoes an addition reaction with a hydroxyl containing reagent in an organic solvent under catalysis, and then hydrolyzing the reaction product and separating to obtain 25-hydroxy cholesterol. The present invention adopts hydroxyl containing reagents such as water, formic acid, acetic acid, propionic acid, butyric acid, benzoic acid, p-methyl benzoic acid to replace the commonly used extremely toxic reagents such as Cr reagent, Hg reagent and polyfluorinated acetone in the prior art. The raw materials of the present invention are easily available, and have low effect on environment. The operation and post treatment are convenient. Moreover, the method has the advantages of mild reaction conditions, simple operation, good selectivity, high efficiency, high yield, simple post treatment, easy product separation, less three wastes and easy industrialization.

The present invention relates to a 25-hydroxy-7-dehydrocholesterol synthetic method, a 25-hydroxy-7-keto-cholesterol derivative is used as a raw material for mechanical milling reaction in a ball mill pot under the effect of sulfonyl hydrazide and a base in the absence of an organic solvent, and then hydrolysis reaction is performed to obtain 25-hydroxy-7-dehydrocholesterol synthetic method. According to the method, in the absence of a solvent, mechanical milling reaction is performed, no dust is spilled over in the grinding process, reaction controllability is good, reaction efficiency is high, the solvent recovery operation in the post-processing process can be avoided, operation is simplified, environmental pollution is reduced, the reaction selectivity is good, yield is high, operation is simple, cost is low, post-treatment is simple, three-waste is less, product is easy to separate, purity is high, the yield is up to 89%, the melting process is short, and the product purity is high.

The invention provides clone and expression aiming at 7,8-dehydrogenase gene. The gene encoding 7,8-dehydrogenase is acquired from a constructed drosophila cDNA library, wherein the 7,8-dehydrogenase can transform cholesterol into 7-dehydrocholesterol, thereby providing a new method for synthesizing the 7-dehydrocholesterol by a biological process, acquiring a process route capable of shortening vitamin D3, reducing production cost, and reducing environmental pollution.

The present invention discloses a method for synthesizing 25-hydroxy cholesterol. The method is as below: subjecting 24-dehydrocholesterol derivative as a raw material, which undergoes an addition reaction with a hydroxyl containing reagent in an organic solvent under catalysis, and then hydrolyzing the reaction product and separating to obtain 25-hydroxy cholesterol. The present invention adopts hydroxyl containing reagents such as water, formic acid, acetic acid, propionic acid, butyric acid, benzoic acid, p-methyl benzoic acid to replace the commonly used extremely toxic reagents such as Cr reagent, Hg reagent and polyfluorinated acetone in the prior art. The raw materials of the present invention are easily available, and have low effect on environment. The operation and post treatment are convenient. Moreover, the method has the advantages of mild reaction conditions, simple operation, good selectivity, high efficiency, high yield, simple post treatment, easy product separation, less three wastes and easy industrialization.

The invention discloses saccharomyces cerevisiae engineering bacteria for producing campesterol and a construction method, and belongs to the field of genetic engineering and bioengineering. An expression cassette element of 7-dehydrocholesterol reductase is introduced into a yeast cell body, and is integrated into a yeast genome by utilizing a CRISPR / Cas9 gene operating system, C-22 sterol desaturase is knocked out, competition of an ergosterol branch path is relieved, and yeast synthesis from glucose to campesterol is realized. The method is simple in process, green and environment-friendly, and can be used for producing campesterol through fermentation. The yield of campesterol prepared by adopting the saccharomyces cerevisiae engineering strain BY4742-delta erg5-TEF1p-dhcr7-ADH1t provided BY the invention can reach 253.35mg / L in a shake flask stage, and the horizontal yield of a fermentation tank can reach 916.88mg / L, which is increased by 2.6 times compared with the yield of campesterol in the shake flask stage of the strain.

The present invention relates to a PEGylated 7-dehydrocholesterol derivative, and a composition for wrinkle alleviation and antiaging, containing the same. The PEGylated 7-dehydrocholesterol derivative, according to the present invention, has improved water solubility and stability, and thus can be effectively used, as an excellent vitamin D source, in a cosmetic composition, a pharmaceutical composition, a dietary supplement and the like for wrinkle alleviation and antiaging.

The invention discloses a preparation method of vitamin D. The preparation method is characterized in that the method comprises the following steps: dissolving provitamin D in an organic solvent to obtain a solution as a raw material, and carrying out a two-step photocatalytic reaction process, separating and purifying to obtain the vitamin D, wherein the provitamin D is 7-dehydrocholesterol, ergosterol, 22, 23-dehydroergosterol, 7-dehydro-beta-sitosterol, 7-dehydro-stigmasterol or 7-dehydro-campesterol, a lamp used in the first-step photocatalytic reaction is an LED ultraviolet lamp with thewavelength range of 250-260nm, and the lamp used in the second-step photocatalytic reaction is an LED ultraviolet lamp with the wavelength range of 350-380nm. The preparation method of the vitamin D is high in yield.

The present invention provides, in certain aspects, novel 7-dehydrocholesterol (7DHC) derivatives that are useful in treating or preventing cancer, as well as in treating or preventing uncontrolled angiogenesis, in a subject. In certain embodiments of the present invention, the subject is a human. In other aspects, the present invention provides a method of preparing compounds of the invention, or a salt or solvate thereof.

The invention relates to the technical field of purification of intermediates, in particular to a method for purifying 24-dehydrocholesterol, which comprises the following steps: (1) taking a mixture containing a compound as shown in a formula (I) and a compound as shown in a formula (II) as a raw material to be purified, and obtaining a first purified product in a mode of step (1A) or (1B); (1A) in the presence of a first solvent, contacting the raw material to be purified with MXn to obtain a complex, and then decomplexing in the presence of water and a second solvent; (1B) carrying out column chromatography separation on the raw material to be purified; (2) crystallizing the first purified product by using a third solvent to obtain a second purified product, wherein R in the formula (I) and R in the formula (II) are the same and are H or C1-C4 acyl groups. The method provided by the invention has the advantages of low cost, simple technical process and capability of effectively purifying 24-dehydrocholesterol.

The present invention discloses a method for synthesizing 25-hydroxy cholesterol. The method is as below: subjecting 24-dehydrocholesterol derivative as a raw material, which undergoes an addition reaction with a hydroxyl containing reagent in an organic solvent under catalysis, and then hydrolyzing the reaction product and separating to obtain 25-hydroxy cholesterol. The present invention adopts hydroxyl containing reagents such as water, formic acid, acetic acid, propionic acid, butyric acid, benzoic acid, p-methyl benzoic acid to replace the commonly used extremely toxic reagents such as Cr reagent, Hg reagent and polyfluorinated acetone in the prior art. The raw materials of the present invention are easily available, and have low effect on environment. The operation and post treatment are convenient. Moreover, the method has the advantages of mild reaction conditions, simple operation, good selectivity, high efficiency, high yield, simple post treatment, easy product separation, less three wastes and easy industrialization.

The invention discloses a recombinant yeast engineering strain for producing 7-dehydrocholesterol and application of the recombinant yeast engineering strain, and belongs to the technical field of genetic engineering and biological engineering. According to the invention, endogenous genes ERG5 and ERG6 of saccharomyces cerevisiae are knocked out, delta (24)-sterol reductase from Gallus gallus is expressed, synthesis of 7-dehydrocholesterol is promoted by regulating expression of genes of endogenous related pathways, the yield of 7-dehydrocholesterol after fermentation for 72 hours is 440.92 mg / L, and the yield of 7-dehydrocholesterol after fermentation for 96 hours is 520.03 mg / L, so that a foundation is laid for subsequent biosynthesis of steroid compounds; the method has potential value and significance for the development of synthetic biology.

The invention belongs to the field of gene engineering, and particularly relates to a prawn Y-organ 7,8-dehydrogenase gene and application thereof. The invention provides a gene for coding 7,8-dehydrogenase, which is obtained from a constructed Metapenaeus ensis Y-organ cDNA (complementary deoxyribonucleic acid) library; and the nucleotide sequence of the gene is disclosed as SEQ ID NO.1. The application method comprises the following steps: cloning the 7,8-dehydrogenase coding gene, and connecting with an expression plasmid to construct a recombinant expression plasmid; after linearizing and purifying the recombinant expression plasmid, converting a receptor bacterium, and screening to obtain a gene engineering bacterium; and carrying out induction expression, purification and identification to obtain the 7,8-dehydrogenase of which the amino acid sequence is disclosed as SEQ ID NO.2. The 7,8-dehydrogenase can be used for converting cholesterol into 7-dehydrocholesterol by a one-step enzyme process, thereby enhancing the conversion efficiency, shortening the process route for producing vitamin D3, lowering the production cost and reducing the environmental pollution.

The invention belongs to the field of inorganic luminescent materials, and particularly relates to lamp powder generating ultraviolet rays B as well as a preparation method and application thereof. The lamp powder is alkaline-earth metal boron chloro aluminate, the lamp powder is applied to a lamp tube, under the excitation of 254 nm ultraviolet rays, the lamp powder can generate the ultraviolet rays B with the wavelength of 297 nm to illuminate the skin, so that subcutaneous 7-dehydrogenated cholesterol can be converted into vitamin D3, the metabolism of calcium and phosphorus of the human body can be promoted, and an effect for preventing children rickets and adult osteoporosis can be realized. In addition, the lamp powder does not contain harmful substances to the human body and is highin safety.

The invention discloses a food vitamin enhancer and a preparing method thereof. The food vitamin enhancer can be obtained by stirring and then drying ergosterol, cyclohexane, livers, isopropanol, milk, 7-dehydrocholesterol, yolk, a neutralizer, soybean oil, phosphoric acid, yeast, magnesium oxide, chloroform and auxiliaries. The melting point of the product is 100-120 DEG C. The weight loss on drying is 0.1-0.3%, and the product comprises 0.01-0.1% of residue on ignition, 0.001-0.003% of heavy metal and 0.0001-0.0003% of arsenic on the basis of the content of 98-105%.