Method of extraction of alkaloids
(57) Abstract:The invention relates to a method by which you can lower cost and with greater purity to obtain alkaloids from plants. Describes the way in which the alkaloid extract or fraction of this extract, he still does not have the form of an aqueous solution, dissolved in water. Set the pH of the aqueous alkaloid extract to alkaline. Then carry out the extraction liquid in a liquid with simple ether and separate the organic phase from the aqueous phase. In the organic phase is almost completely alkaloid-lupain, while in the aqueous phase are present in the original extract, alkaloids. 13 C.p. f-crystals. The invention relates to a method of extraction of alkaloids, especially lupinine of alkaloids.steroid plant restrictive part of paragraph 1 of the claims.The prior art is still known to produce alkaloids extraction, for example, from flour obtained from the seeds of the plants with an organic solvent. While in the literature as extractants offer simple diethyl ethers or chlorinated hydrocarbons, such as chloroform or dichloromethane. Podobna, that simple ethyl esters have the tendency to form peroxides, which required large maintenance costs and equipment costs. To prevent such explosive peroxides add stabilizers simple ether. But they pollute the extracts, which is considered a disadvantage. Therefore, the purity of the obtained alkaloids is poor.Further, it is known, especially to reduce the bitterness of bitter Lipunov, obtaining the so-called water alkaloid extracts. Such water alkaloid extracts can be obtained in various ways. Example to obtain such water alkaloid extract disclosed in EP 0084547. In this patent the seeds of the lupine grind to flour with a specific grain size and then the flour in various technological operations is mixed with water and again separated.Proceeding from the aforementioned prior art, the present invention is to propose ways by which you can simply, with less cost and high purity to obtain alkaloids from plants.This problem is solved thanks to the distinctive characteristics of paragraph 1. The result is given in the dependent claims operations docno invention lies an unexpected discovery that lupain in pure form can be distinguished from simple esters from the aqueous alkaloid extract by the method of extraction liquid in the liquid.Therefore alkaloid extract according to the invention, since it is not presented as an aqueous solution, dissolved in water. Water alkaloid extract set at the alkaline pH value. If alkaloid extract has previously been dissolved in another way, you can also use hence the resulting fraction of this alkaloid extract. Then spend named the extraction fluid in the fluid with simple ether and separate the organic phase from the aqueous phase. Now in the organic phase is almost completely alkaloid lupain, while in the aqueous phase contains other available in the original extract alkaloids.From now presents the organic phase can be obtained respectively lupain, while the water phase can be used to obtain at least another, different from lupinine alkaloid.A particularly large output lupinine or particularly good separation lupinine from the aqueous phase is possible at pH values in the range of 10 to 13.5 during extraction. Very good results were achieved in the woods should not be used acid-resistant materials.As a simple ether used is preferably a simple tert.-butyl methyl ether (tBME). Special preference tBME before the usual simple diethyl esters is that this simple ether has no tendency to the formation of peroxide and as a result not a cause for concern with the technical application. Thanks to this process easier and, consequently, also more economical.Preferably to highlight lupinine containing loanin the organic phase to evaporate at least partially, for example, in a vacuum extractor, so that the organic extract at least concentrate.In a particularly pure form, you can get lupain from the organic phase, in which loanin or preferably narrowed organic extract is cooled until, until crystallizes loanin. In addition to the described high purity, this method provides the advantage that in addition to water and simple ether as the extracting agent does not use any other solvents to highlight lupinine. The fact that lupain can be obtained in crystalline, molecular form without the other operations directly from the extractant, is the following neozed is stylizacji lupinine recommended the addition of crystallization centers.Already remarkably high degree of purity obtained in this way lupinine increases, and the resulting crystals are recrystallized, preferably again in tM. Obtained in this way lupain has a purity of > 98%.As already mentioned, the aqueous phase remaining when conducted according to the method of the invention the extraction liquid in the liquid, can also be mined to get at least another lepinoy alkaloid. Here take into account, in particular, alkaloids sparteine, 13-oxylipin, 3-oxylipin, 17-oxoguanine, n-medillin, angustifolii, tetrahydrobiopterin, lupinine, multiflori, Albin, ammodendron and alkaloids of ester formed from 13 - and 3-oxylipin.Now for the further identification of such alkaloids aqueous phase from the extraction according to the method of the invention, the liquid in the liquid set to a pH value of more than 12.5. Especially good results were achieved in the range of pH-values between 13 and 13.5. Now again conduct the extraction liquid in a liquid organic solvent. For use as solvents are particularly suitable was chlorinated hydrocarbons and particularly dichloromethane or chloroform, is here the organic phase can also separate other alkaloids, for example, chromatography, and should encourage the preceding contraction, for example, in a vacuum. For the separation we should mention especially the use of so-called instant chromatography, the application of which in this regard still was not known.Obtaining different from lupinine alkaloid can be made through distillation. This method can be applied, in particular, directly to the aqueous phase. First of all you can select by distillation alkaloid sparteine.As starting material for the present method is suitable all alkalotolerant plants, especially plants with alkaloids finalizada and especially their seeds. These plants are mainly in the field pea, among which should again highlight lupine, for example, Lupinus albus (white Lupin), Lupinus luteus (yellow Lupin), Lupinus mutabilis, Lupinus angustifolius (narrow-leafed Lupin), Lupinus polyphyllus (overlapping lupine), Lupinus varius, Lupinus micrantus or Luinus cosentinii. The high number lupinine contain Lupinus albus, Lupinus angustif olius and Lupinus mutabilis. Next, Lupinus luteus should mention great content sparteine. Other alkaloids found in various combinations in all lupine, primarily in the so-called bitter it must however be below 0.05 wt.%.Rich luprinol extract as the source of the extract method according to the invention can be obtained through supercritical CO2extraction from plants, especially from the flour of the seeds of plants, which should be processed further, as described, by way of the extraction liquid in the liquid.An example embodiment of the invention explained in detail on the basis of the following description.As a starting point can be obtained by applying cited in the prior art EP 0084547 water alkaloid extract or above is obtained by CO2-extraction of the extract. However, the original extract can also be obtained through the following extraction flour.Extraction flour
Flour Lupinus albus or Lupinus angustifolius is mixed with water and long mix. This establishes the equilibrium concentration of alkaloids in the flour and liquid. During the initial quantities of 10 kg of flour and 90 kg of water can be obtained after separation of the phases (for example, with the help of decanter or filtering) about 75 kg of an aqueous extract containing alkaloid about 2.5 g/l to get from here to a clear solution, the extract should is this loss solution extract or loss due to the separation of solids comprise between 15 and 25%. For extraction of the fluid in the fluid use about 60 kg of extracts containing alkaloids 2.5 g/l In the extract contains 150 g of alkaloids.Selection lupinine
The extraction liquid in a liquid
This aqueous extract using sodium liquor set to a pH value of about 11 and extracted with simple tert.-butylmethylamine ether (tBME) in the ratio of 1:1 (volume/volume). When multistage extraction of loanin almost completely transformed into the organic phase. For a constant pH-value during the extraction of the required pH-regulation, so as to increase the extraction pH-value drops and leaves for optimal extraction lupinine range. Consequently 105 g lupinine and even the rest of 14-15 g other alkaloids can be extracted in the organic phase. The organic extract contains about 120 g of alkaloids with the number lupinine from 88 to 90%. Aqueous extract after extraction contains about 30 g of alkaloids with number 13-oxylipin above 50%.The organic extract
The organic phase is concentrated to approximately 1/100 of its original volume (e.g., by rotary evaporation apparatus in a vacuum of about 100 mbar, T = 40oC), and obtained in this way is to -12o. At this time of the solution precipitates already white crystals lupinine. The process can be accelerated by the persecution of the cooled solution with crystal lupinine.Obtained, for example, by decantation of the solvent, the crystals contain about 93% lupinine (94,5 g) and about 7% of impurities (about 7 g). In the organic supernatant are about 10% of the total lupinine and residual alkaloids and other substances (dyes).Recrystallization
The separated crystals are dissolved with approximately 3 - to 5-fold amount of weight in tBME and again cooled. After a new crystallization, phase separation, simple ether and determine the purity of lupinine the latter has a purity > 98%. The total number of detachable lupinine is about 85 grams, which corresponds to a yield of about 80%.Repetition of this procedure, you can achieve pureness of above 99%. By recrystallization or washing of the crystals losses lupinine be about 20%.Selection 13-oxylipin
Aqueous extract after the extraction lupinine differentiates 13-oxylipin (contains about 15 g of 13-oxylipin). For this pH-value is set to >12.5 and extracted with dichloromethane in Rel is torono distilled off. The remaining oily concentrate is served on a column of silica gel.Due to the varying means to elution with increasing polarity (toluene ---> tBME ---> CHCl3--> CH2Cl2---> CH2Cl2/MeOH) one after another wash out impurities and dyes from the column, so that, finally, with a mixture of CH2C2/methanol can be obtained relatively pure fraction 13-oxylipin.After separation of the solvent in this way it is possible to allocate about 10 g of 13-hydroxy-lupinine as oily liquid (purity > 80%).Accordingly it is possible to allocate the remaining alkaloids Lupino, and particularly suitable turned out to be an instant chromatography. 1. Method of extraction of alkaloids, in particular lupinine, alkaloids.steroid plants such as lupine, by simple extraction with ether, characterized in that the aqueous extract of alkaloids from alkaloids.steroid plants, parts of such plants, in particular of their seeds or flour, and bring to alkaline pH values, b) the extraction is carried out, the liquid in the liquid with a simple ester, and C) separating the organic phase from the aqueous phase.2. The method according to p. 1, characterized in that lupain separated from the body of the 2, characterized in that the pH value under method (a) is in the region of 10 to 13.5.4. The method according to one of paragraphs.1 to 3, characterized in that the pH value under method (a) is in the area of 10,5 - 11,5.5. The method according to one of paragraphs.1 to 4, characterized in that a simple live on stage method b) represents a tert.-butyl methyl simple ether.6. The method according to one of paragraphs.1 to 5, characterized in that the organic phase is at least partially evaporated.7. The method according to one of paragraphs.1 - 6, characterized in that the organic phase is cooled at least as long until crystallizes loanin.8. The method according to one of paragraphs.1 to 7, characterized in that the crystallization lupinine add the crystallization centers.9. The method according to one of paragraphs.1 to 8, characterized in that lupain purified by recrystallization.10. The method according to one of paragraphs.1 to 9, characterized in that the aqueous phase in an alkaline medium is subjected to further extraction liquid in a liquid organic solvent to highlight 13-hydroxydopamine away from the resulting organic phase.11. The method according to p. 10, characterized in that the pH value of the extraction is boob on PP.10 - 11, wherein the organic solvent is a chlorinated hydrocarbon.14. The method according to p. 12, characterized in that the organic solvent is dichloromethane.
SUBSTANCE: there is recovered a fragment of genomic DNA Pseudomonas fluorescent A2-2, including full-size gene cluster of biosynthesis of safracin (A and B) analysis of which showed the presence of several "OPC" arranged in two operons: sacABCDEFGH and sacIJ. Expression of nucleic acid containing full gene cluster of safracin in a heterosystem enabled producing recombinant forms of natural safracins A and B.
EFFECT: removal or malfunction of separate genes found in operons, and applications of the produced modified forms of nucleic acid in the recombinant DNA technology have resulted in synthesised analogues of safracin to be used as an antimicrobial or antitumour agent, and also in synthesis of ecteinascedine compounds.
20 cl, 9 dwg, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: there are described new N-cyclic sulphonamido-compounds and their pharmaceutically acceptable salts of formula : of formula 1a of formula 1b of formula 1c where the ring A means phenyl, thienyl which can be substituted by halogen, or pyridinyl; a value of the ring B is presented in the patent claim, and also a pharmaceutical composition containing them, and a method of treating Alzheimer's disease.
EFFECT: compounds are gamma-secretase inhibitors and can be used for treating Alzheimer's disease.
16 cl, 98 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to fluorinated catharantine derivatives of general formula I in which broken line represents possibility of double bond presence, when substitution X is absent, or simple bond, when X stands for substitution with another group: H, R1, R2 and R3 independent on each other represent a fluorine atom or methylated group, and n=2.
EFFECT: invention also relates to method of obtaining said derivative and to its application as intermediate compound for synthesis of fluorated dimeric Vinca alkaloids, in particular vinflunine, which in its turn is used as anti-cancer agent.
SUBSTANCE: invention relates to improved method of obtaining pyridine compounds (AA),(BB) and (CC) of respective formulas:
said compounds possess inhibiting action with respect to HIV-integrase. method consists in carrying out the following stages: P-1) bromination of compound of formula (I-I) with obtaining bromine-compound of formula (II-II)
where value R represents -CHO, -CH(OH)2, -CH(OH)(OR4), -CH(OH)-CH2OH or -CH(OR5)(OR6); P1 represents benzyl; P3 represents H or protective group of carboxyl; R4 represents lower alkyl; R5 and R6 independently represent lower alkyl or R5 and R6 can represent alkyl and be connected with formation of 5-, 6- or 7-member ring, P-2) formation of side chain of 2,4-di-fluorophenyl-CH2-NH-C(O)- with application of reagents 2,4-di-fluorophenyl-CH2-NH2 and carbon monoxide, stage of formation of Q ring by means of respective amine, selected from 3-amino-butan-1-ol, 2-amino-propan-1-ol and 2-pyrrolidinyl methylamine, and stage of debenzylation with obtaining compound of formula (AA), (BB) or (CC), where said stage P-2 is carried out after formation of Q ring.
EFFECT: method makes it possible to simplify obtaining target compounds due to carrying out regioselective bromination at the first stage.
6 cl, 3 ex, 7 dwg
SUBSTANCE: invention relates to field of organic chemistry, namely to method of obtaining 4a,5b,10,12-tetraazaindeno[2,1-b]fluorene, which consists in the fact that interaction of pyridine with 1,5-dichloro-2,4-dinitrobenzene is carried out at temperature 20°C in acetone and molar ratio 1,5-dichloro-2,4-dinitrobenzene:pyridine = 1:4 for 4 hours, reduction of alcohol salt solution 1,1'-(4,6-dinitro-1,3-phenylene)dipyridinium is realised with solution SnCl2·2H2O in 3% hydrochloric acid and molar ratio salt 1,1'-(4,6-dinitro-1,3-phenylene)dipyridinium : SnCl2·2H2O = 1:6 at temperature 40°C for 0.25 h.
EFFECT: method of binding, which is characterised by reduction of synthesis time and increase of purity and target product output, has been elaborated.
SUBSTANCE: invention relates to compound of formula I or its pharmaceutically acceptable salts, which are inhibitors of Trk kinases and are useful for treating pain, malignant oncological disease, inflammation, neurodegenerative diseases and Trypanasoma Crusi infections. Invention also describes versions of methods of producing compound of formula I. In Formula I (I) ring A is selected from A-1, A-2, A-3, having structure , where wavy line marked 1 indicates connection point of ring A to ring B, and wavy line marked 2 indicates point of connection ring A to W; X is N or CH; Y is H or F; R1 is H or halogen; ring B is selected from rings B-1 and B-2, having structure , where wavy line marked 3 indicates point of connection to ring A, and wavy line marked 4 indicates point of connection to pyrazolo[1,5-a]pyrimidine ring of formula I; W is O, NH or CH2, wherein, when ring A is A-2, then W is CH2; m is 0, 1 or 2; D is carbon, R2 and R2a independently represent H, F, (1-3C)alkyl or OH (provided that R2 and R2a are not simultaneously OH), and R3 and R3a are independently H or (1-3C)alkyl, or (D) is carbon or nitrogen, R2 and R3 are absent, and R2a and R3a together with atoms to which they are bonded, form 5-6-member heteroaryl ring containing 1-2 heteroatoms ring; Z is *-NR4aC(=O)-, *-ONHC(=O)-, *-NR4bCH2- or *-OC(=O)-, where asterix indicates connection point of Z to containing carbon R3; R4a is H or (1-6C)alkyl; R4b is H, (1-6C)alkyl, ((1-6C)alkyl)C(O)-, HOCH2C(O)-, ((1-6 C)alkyl) sulfonyl, HO2CCH2- or ((1-6C)alkyl)NH(CO)-; and R-5 and R6 independently represent H, halogen, OH or (1-6C)alkyl.
EFFECT: useful for treating pain, malignant oncological disease, inflammation, neurodegenerative diseases and Trypanasoma Crusi infections.
73 cl, 1 tbl, 45 ex
SUBSTANCE: invention relates to a compound of general formula
or , or pharmaceutically acceptable salts thereof, R is methyl, ethyl, propyl or benzyl; * - X - * is selected from the group consisting of , and ; Y is (-CH2-)3 or (-CH2-)4. The invention also relates to a method for preparation of compounds of general formula 1 or 2.
EFFECT: new compounds are obtained that have the properties of protein-protein interactions inhibitors.
4 cl, 2 tbl, 3 ex