The method of obtaining 18-carboxy-20-(carboxymethyl)-8-ethynyl-13 - ethyl-2,3-dihydro-3,7-12,17-tetramethyl-n, norfin-2 - propionic acid or its salts
(57) Abstract:Usage: biotechnology, chemical and pharmaceutical industry. The inventive receiving 18 carboxy-20(carboxymethyl)-8-ethynyl-13 - ethyl-2,3-dihydro-3,7,12,17 - tetramethyl-N,N porphin-2 - propionic acid or its salts. Getting chlorin e6carry out when using as a raw material biomass of cyanobacteria, for example, of the genus Spirulina, obtained by cultivation in the conditions of artificial or solar light on a nutrient medium having a pH of 8.0 to 12.0, with pre-treatment of biomass alcoholic solution of alkali. The resulting alcoholic extract of chlorophyll treated with acid to obtain pheophytin a. Pheophytin washed with a nonpolar solvent such as hexane and subjected to saponification to the formation of the target product, which was precipitated by treatment with hydrochloric acid. 4 C.p. f-crystals. The invention relates to organic chemistry, in particular to an improved process for the preparation of porphyrins, in particular to a method for producing chlorin e6(18-carboxy-20-(carboxymethyl)-8-ethynyl-13-ethyl-2,3-dihyd - ro-3,7,12,17 - tetramethyl-n, n. porphin-2-propionic acid) or its salts, which can find application in chemical farmacity the basis of chlorophyll . It is known that vegetable porphyrins, including chlorophyll and its derivatives are pharmacologically active compounds and can be used in medical practice, including as photosensitizers for photodynamic diagnosis and therapy of oncological diseases. Currently known methods for producing chlorin e6based on chemical transformations of chlorophyll in the target product.In particular, the most common method of producing chlorin e6by processing the chlorophyll extract derived from soybeans, 30% hydrochloric acid with a mixture of pheophorbide and which is then treated with 0.5% methanol solution of alkali in a vacuum. The target product is removed from the reaction mixture with ether.There is also known a method of producing chlorin e6from nettle leaves. For this nettle leaves pour ethanol. The result: an extract containing a mixture of chlorophylls and . The extract is treated with concentrated hydrochloric acid, resulting from each form of chlorophyll contained in the extract, is formed corresponding to the shape of pheophytin and . Subsequent alkaline saponification of different forms of pheophytin PR is osenia these products is 4:1, respectively. Along with these major products in the reaction mixture present their esters, as well as ficinia remains.To highlight chlorin e6of a mixture of products, the reaction mixture is first neutralized with concentrated hydrochloric acid to pH 7.0, and then add additional concentrated hydrochloric acid in an amount corresponding hydrochloric number of chlorin e6. Thus from the reaction mixture extract 2/3 dissolved chlorine e6.Consumption of concentrated hydrochloric acid for cleaning of chlorin e6120 ml per 1 l of the reaction mixture, and just to receive 1 g of chlorine consume 2.5 liters of concentrated hydrochloric acid.At the specified way we obtain 1 g of chlorin e6from 1 kg of dried nettle leaves, with the purity of the product is from 30% to 60% of the basic substance. For more pure target product using the method of column chromatography of alumina (eluent-methylene chloride:benzene (2: 1)).Thus, in known methods for producing chlorin e6in the first stage of the technological cycle are used extracts of chlorophyll obtained from chloropalladite parts of higher plants or Zelenyi the ratio of the individual forms of chlorophyll at different plants varies and depends on the adaptability of plants to different light conditions. "Sun-loving" plants the ratio of chlorophyll in the leaves is approximately 3:1, "shade" it is decreased.When processing green plants organic solvents retrieves all forms of chlorophyll ( and ) and the subsequent processing of these extracts is formed or a mixture of pheophorbide and (first method), or a mixture of pheophytins and (as noted in the second method, which ultimately leads to the formation of a mixture of chlorin e6and the birthplace of the g7.The disadvantages of the known methods for producing chlorin e6are the complexity of the technological process, the use of large volumes of concentrated hydrochloric acid (2.5 l per 1 g of chlorin e6), a small yield and low purity of the target product (from 1 kg of dried nettle leaves receive 1 g of chlorin e6, the purity of which is not higher than 60%), the use of chromatographic method for purification of the target product. In addition, given the complex environmental conditions, does not exclude the possibility of contamination of vegetable raw materials of pesticides and other toxic chemicals, and the use of spontaneously grown biomass of green plants as a source of chlorophyll can lead ultimately to the pollution of Mecca, significant reduction of corrosive hydrochloric acid and obtain the desired product with a purity not less than 90% without the use of chromatographic purification.This is achieved by the proposed method of producing chlorin e6implying that as a source of raw materials use biomass of cyanobacteria usually of the genus Spirulina, mainly grown in the light in controlled conditions on an artificial nutrient medium having a pH ranging from 8.0 to 12.0. The cyanobacteria biomass treated with an aqueous-alcoholic solution of alkali, mainly when the ratio of water:alcohol of 1 to 2-3 and the concentration of alkali 10-2-10-3N. filtered and extracted chlorophyll "a" ethyl alcohol. The combined organic extracts are treated or hydrochloric acid formed pheophytin filtered off, washed with a nonpolar organic solvent, usually hexane.Alkaline hydrolysis of pheophytin carried out in the absence of oxygen in the alcohol medium when heated. The formation of the alkali salt of chlorin. After cooling, the reaction mixture was adjusted with hydrochloric acid to a pH of 6.0-7.0 (consumption of acid 50-55 ml per 1 l of the reaction mass, obrazovanii transferred to salt conventional techniques.The use of the proposed method can improve the yield of the target product at least 3 times to obtain a product with a purity of not lower than 90% in Addition, the consumption of concentrated hydrochloric acid was reduced approximately 10 times.To remove water-soluble proteins, hydrocarbons and other impurities, the cyanobacteria biomass pre-treated with an aqueous-alcoholic solution of alkali, for example, when the ratio of water and alcohol of 1 to 2-3 and the alkali concentration of 10-2-10-3N. To increase the yield of the target product, it is expedient according to the claimed invention to use the biomass of cyanobacteria containing (dry residue) of 2.0 to 4.5 wt. chlorophyll forms , 46-62 wt. proteins, 4-9 wt. lipids, 10-20 wt. carbohydrates, 10-12 wt. ash elements.Further objectives and advantages of the invention will become apparent from the subsequent detailed description of the method of producing chlorin e6and specific examples of implementation of this method.In accordance with the claimed invention receive 18-carboxy-20-(carboxymethyl)-8-ethynyl-13-ethyl-2,3-dihydro-3,7,12, 17 - tetramethyl-n.N. porphin-2-propionic acid, called chlorin e6carry out from a substance containing x is lnyh environments in terms of solar or artificial light.Monoculture of cyanobacteria grown in a special fermenters closed, eliminating the possibility of green algae, and therefore getting into the biomass of chlorophyll forms .Use chloropalladite plant materials standard composition grown in algological and clean environment ensures high purity of chlorin e6free from traces of heavy metals and other impurities caused by pollution. This fact is especially important in the manufacture of medical products.The biomass of cyanobacteria, it is advisable to handle hot water-alcohol solution of alkali, for example, at a temperature of 40-60aboutC, the ratio of water and alcohol of 1 to 2-3 and the alkali concentration of 10-2-10-3N. When this treatment is the destruction of cell membranes, removed low molecular weight fractions of carbohydrates and soluble proteins, which makes it difficult for subsequent extraction of chlorophyll and contaminate the extract, and therefore, the target product. Biomass after processing is extracted with ethyl alcohol, extracting the chlorophyll . On polucheniya in the reaction mass pheophytin , which is marked by filtration.Received pheophytin , the output of which is 10 g of 1 kg of biomass of cyanobacteria, washed with a nonpolar solvent such as hexane, to remove carotenoids and lipids from the surface of the crystals pheophytin .Then cleaned pheophytin dissolved and subjected to saponification with alkali in the absence of oxygen, for example, in a stream of nitrogen. In the reaction mass is formed target product chlorin e6. To highlight chlorin e6the reaction mixture is neutralized with hydrochloric acid, taken according to the invention in quantities of 50-55 ml per 1 l of the reaction mixture until crystals deposition of chlorin e6that is separated by filtration.Thus, the proposed method allows to obtain 1 kg of biomass of cyanobacteria 3.5 g of chlorin e6with a purity of at least 90% at a flow rate of concentrated hydrochloric acid in the amount of 220 ml per 1 g of the target product, which is 10 times less than the amount of acid used in the method-analogue.P R I m e R 1. The biomass of cyanobacteria of the genus Spirulina is grown in a closed glass fermenter under conditions of sunlight or artificial lighting on liquid nutrient medium, representing Vodniy. The culture medium has a pH ranging from 8.0 to 12.0. The carbon source is carbon dioxide, periodically introduced into the nutrient medium. After reaching the suspension of certain cell concentration (2.5-3.0 g/l) biomass of cyanobacteria is separated from the nutrient medium by filtration. 1 kg of the thus obtained biomass of cyanobacteria with a humidity of 75-80% pour 2.5 liters heated to 60aboutWith alkaline ethanol solution obtained by dissolving 4 g of NaHCO3in 1 l of distilled water and subsequent mixing with ethanol in the ratio 1:3, respectively.The mixture of biomass with the specified solution is stirred for 3-5 min, and then filtered. Biomass on the filter is shaken out three times with ethyl alcohol, extracts containing chlorophyll , summarize and treated with glacial acetic acid at the rate of 1.5 ml per 1 l of the extract. The resulting crystals pheophytin filtered and washed with hexane. The output of pheophytin is 10,Pheophytin in an amount of 10 g was dissolved in 3 l of alcohol by boiling in a stream of nitrogen, add 400 g of KOH and carry out the saponification reaction for 10 minutes then the mixture is neutralized with a solution containing 45 ml of concentrated soles filtered. The crystals are dried in the air without light. The purity of the obtained product 92%
P R I m m e R 2. Use the cyanobacteria biomass obtained in conditions similar to those specified in example 1.1 kg of the above biomass of cyanobacteria pour 1.5 liters of alkaline ethanol solution, which is obtained by dissolving 40 ml of 1 N. NaOH in 1 l of distilled water and subsequent mixing with ethanol in a ratio of 1 to 2, respectively.The mixture of biomass with the specified solution is stirred for 3-5 min, filtered, biomass extracted three times with ethyl alcohol to extract the chlorophyll . The extracts are combined and add 1.2 ml of concentrated HCl per liter of extract. The resulting crystals pheophytin filtered and washed on the filter with hexane. The output of pheophytin is 10 g 10 g pheophytin dissolved in 3.3 liters of ethyl alcohol boiling, then add 400 g of KOH and carry out the saponification reaction for 10 min in vacuum. After that the reaction mixture is neutralized with hydrochloric acid to pH 6-7. Consumption of concentrated hydrochloric acid is 55 ml.Crystals of chlorin e6from the reaction mixture is filtered and dried. Weight obtained Karlovich, such as the one outlined in example 1, when the cultivation of the strain of Spirulina species Fr-A 127.Alcohol extract of chlorophyll receive, as described in example 1. To 1 liter of the extract add 2 ml of 1.0 N. oxalic acid. The resulting crystals pheophytin filtered and washed on the filter with hexane. The output of teofilina of 9.7 gReceived pheophytin omelet, as described in examples 1 and 2. The output of chlorin e6is 3.3 g with a purity of 90% 1. The METHOD of OBTAINING 18-CARBOXY-20-(CARBOXYMETHYL)-8-ETHYNYL-13-ETHYL-2,3-DIHYDRO-3,7-12,17-TETRAMETHYL-N, NORFIN-2-PROPIONIC ACID OR ITS SALTS, including extraction of chlorophyll from plant materials, treatment of the extract with acid, followed by alkaline hydrolysis and isolation of the target product in the form of acids or salts, characterized in that the feedstock used the biomass of cyanobacteria, which is pre-treated with an aqueous-alcoholic solution of alkali, after treatment of the extract chlorophyll acid product obtained is filtered off and washed with a nonpolar organic solvent and subjected to alkaline hydrolysis in a vacuum.2. The method according to p. 1, characterized in that is used as raw material of biobattery of the genus Spirulina, grown on the light under controlled conditions on a nutrient medium having a pH of 8.0 to 12.0.4. The method according to p. 1, characterized in that as the use of alkali hydroxides or carbonates of alkali metals.5. The method according to p. 1, characterized in that as a non-polar organic solvent used hexane.
FIELD: organic chemistry.
SUBSTANCE: invention relates to new derivatives of metalloporphyrazine of the general formula (I): wherein M means Cu, Co. These compounds can be used as dyes, catalysts in different processes and materials of sensitive members of gas sensor.
EFFECT: valuable properties of compounds.
2 cl, 6 sch, 1 dwg, 5 ex
SUBSTANCE: invention relates to method for production of synthetic chlorophyll (Chl) or bacteriochlorophyll (Bchl) derivatives of general formula I , wherein X is O;. Claimed method includes interaction under anaerobic conditions of Chl, Bchl derivatives containing COOCH3-group in C-132-position and COOR3-group in C-172-position in presence of tetraethyl orthotitanate. Further compounds of formula I wherein R1 and R2 are different radicals are obtained in aproton solvent such as peroxide-free tetrahydrofurane and dimethyl formamide, and compounds of formula I wherein R1 and R2 are the same ones are produced by using R1OH as a solvent. Derivatives of present invention are useful as stabilizers, linkage/spacer for binding another acceptable molecules to Chl/Bchl macrocycle.
EFFECT: simplified method for production of various chlorophyll or bacteriochlorophyll derivatives.
13 cl, 3 ex, 2 tbl, 8 dwg
SUBSTANCE: invention relates to method for production of porphyrinopeptides satisfying the formula I , wherein R1 and R2 independently from one another represent amino acids or peptides comprising 2-15 of amino acid residues, wherein α-carboxylic groups of amino acids or peptides may be modified by C1-C8-alkyl ester and side functional groups of amino acids or peptides may be protected; in particular R1 is ArgOMe; R2 is -OH (III); R1 is LeuHisOMe; R2 is -OH (IV); R1 is LeuLeuValPheOMe; R2 is -OH (V); porphyrin carboxylic group may be modified by methyl or other C1-C9-ester or pharmaceutically acceptable salt; Y- represents Cl-; Me represents Zn, Cu, Fe, Mn. Claimed method includes activation of porphyrin carboxylic group with N-oxy-5-norbornene-2,3-dicarboxyimede in molar ratio of 1:1 in presence of N,N'-dicyclohexylcarbodiinide; or with diphenylphosphorylazide (DPPA) in equimolar ratio of porphyrin/DPPA in presence of base. Then porphyrin with activated carboxylic group is brought into reaction with amino component (amino acid or peptide) in form of mineral acid salt, which is neutralized with base. Also disclosed are methods for application of compounds (I) as nucleotic agents.
EFFECT: new nucleotic agents.
4 cl, 7 ex, 1 tbl
FIELD: chemistry of metalloorganic compounds.
SUBSTANCE: invention relates to new derivatives of metalloporphyrazines of the general formula (I):
wherein M means Cu, Co. These compounds can be used as dyes, catalysts of different processes and material for sensitive members of gas pickups.
EFFECT: valuable properties of complexes.
4 fig, 1 dwg, 5 ex
FIELD: organic chemistry, medicine, oncology.
SUBSTANCE: sensitizing agent for photodynamic disruption of malignant neoplasm cells represents a new compound - 18-hydroxy-substituted cycloimide of chlorine p6 or 7,8-dihydroxybacteriochlorine of the formula (I):
. Compounds of the formula (I) are used as an agent in treatment of malignant neoplasm. Invention expands assortment of sensitizing agents used in carrying out the photodynamic therapy.
EFFECT: valuable medicinal properties of agent.
6 cl, 1 dwg
FIELD: organic chemistry, chemical technology.
SUBSTANCE: invention relates to 2,3-dicarboxy-6,7-dimethylanthraquinone of the formula (I):
that is characterized by melting point value Tm 207°C. This compound can be used as the parent substance in synthesis of metal complexes tetra(-di-6,7-carboxy)-anthraquinonoporphyrazine used as dyes and catalysts. Also, invention describes methods for synthesis and using compound of the formula (I).
EFFECT: value properties of compound and complexes.
4 dwg, 6 ex
FIELD: organic chemistry.
SUBSTANCE: invention relates to 2,3-dicarboxy-5,6,8-trimethylanthraquinone of the formula (I):
. This compound can be used in synthesis of metal complexes of tetra-(tri-5,6,8-carboxy)-anthraquinonoporphyrazine used as dyes and catalysts. Compound of the formula (I) is characterized by melting point value (Tm) 207°C. Also, invention describes methods for synthesis and using compound of the formula (I).
EFFECT: valuable properties of compound.
4 cl, 6 ex
FIELD: organic chemistry of complex compounds.
SUBSTANCE: invention relates to new derivatives of metalloporphyrazines, namely: to metal complexes of tetra-(tri-5,6,8-methyl)-anthraquinonoporphyrazine of the general formula:
. These compounds can be used as parent substances for synthesis, respectively, copper and cobalt complexes of tetra-(tri-5,6,8-carboxy)-anthraquinonoporphyrazine used as dyes, catalysts in different processes, materials in sensitive members of gas transducers.
EFFECT: valuable properties of complexes.
2 cl, 7 dwg, 8 ex
FIELD: organic chemistry of complex compounds.
SUBSTANCE: invention relates to new derivatives of metalloporphyrazines, namely to metal complexes of tetra-(tri-5,6,8-carboxy) -anthraquinonoporphyrazine of the general formula:
. These compounds can be sued as dyes, catalysts in different processes and materials for sensitive members of gas transducers.
EFFECT: valuable properties of compounds.
2 cl, 5 dwg, 6 ex
FIELD: chemistry of metalloorganic compounds, chemical industry.
SUBSTANCE: invention relates to preparing compounds of tetrapyrazinoporphyrazine order, namely, to cobalt octasulfooctaphenyltetrapyrazinoporphyrazine of the formula:
that can be used as a catalyst in oxidation reactions of sulfur-containing compounds, in particular, cysteine and thioureas, and diethylamine also being both in acid and neutral media.
EFFECT: valuable properties of compound.
2 cl, 2 dwg, 4 ex
FIELD: biotechnology, agriculture, pharmaceutical industry.
SUBSTANCE: claimed method includes Chlorella vulgaris cultivation, double freezing thereof, cell juice separation and solid cut extraction with catholyte. Then obtained extract, cell juice and liquid cut, produced by Chlorella vulgaris cultivation are blended.
EFFECT: bioactive substance complex containing enzymes and antibiotics.
SUBSTANCE: method provides stocking with fish the pond with herbivorous fishes, namely, white and particolored silver carps and addition of the microalgae suspension to the pond (algolization). Method involves using suspension of chlorella of the strain Chlorella vulgaris BIN in the cell density 50-60 x 106 cells/ml in the amount 20-40 l. The suspension is added in 2-4 points in upper and median parts of the pond and for enhancing the effect the pond is stocked with fish with herbivorous fishes, namely, two-aged white and parti-colored silver carps in the amount 1-10 fishes per ha. The proposed protection method of pond prevents water "flowering" with blue-green algae and creates the favorable ecological situation.
EFFECT: improved control method.
1 tbl, 1 ex
FIELD: microbiological industry; equipment for growing of microseaweeds.
SUBSTANCE: the invention is pertaining to microbiological industry, to the equipment for growing of microseaweeds, in particular, to the installation for growing of a chlorella. The installation for growing of microseaweeds, in particular, a chlorella includes a capacity located on a skeleton used for suspension of microseaweeds, in which there are the upright mounted cylindrical glass shells with the stationary placed in them electric bulbs. The capacity is supplied with the fans, installed under the shells and used for the air supply inside the shells at the moment the temperature of the suspension exceeds the optimal temperature of the cultivation. The installation is supplied with a suspension temperature detector located inside the capacity connected with a temperature controller connected to the fans. The invention ensures a reliable control over the temperature during cultivation of microseaweeds and keeping the temperature within the optimal limits for raising the productivity of the installation.
EFFECT: the invention ensures a reliable control over the temperature during cultivation of microseaweeds and keeping the temperature within the optimal limits for raising the productivity of the installation.
FIELD: phytobiotechnology, microbiology, medicine, food processing industry.
SUBSTANCE: method for preparing zinc-enriched spirulina biomass (Spirulina platensis) involves by simultaneous addition of zinc nitrate or sulfate in the concentration 10-30 mg/l and seeding inoculum in the concentration 0.1-0.3 mg of dry mass/l to the cultural medium. Method provides preparing spirulina biomass in the concentration 3.3-4.30 mg/g of dry mass and the yield of biomass 2.5-3.6 g/l for short period (4-5 days of cultivation).
EFFECT: improved preparing method.
4 tbl, 2 dwg, 6 ex
FIELD: agriculture, in particular, presowing treatment of farm crop seeds.
SUBSTANCE: method involves treating seeds with mixture containing biologically active composition and seed disinfectant before sowing. Biologically active composition is suspension of green algae Chlorella Vulgaris having density of 60-80 million cells per ml in an amount of 12-16 l/ton of seeds. Seed disinfectant is preparation corresponding to the given crop, consumption norm making 60-70% of recommended value.
EFFECT: increased yield and biological value of farm crops.
FIELD: biotechnology, in particular process of microalgae culturing.
SUBSTANCE: method for culturing of thermophile cyanobacteria belonging to genus Phormidium includes substrate saturation with carbonic acid, controlling of illumination, temperature and substrate chemical composition. Controlling of illumination, temperature and substrate chemical composition is carried out by feeding of natural thermal water providing flowage thereof.
EFFECT: simplified culturing method with decreased energy consumption.