Method for preparing betulinic acid

FIELD: natural compounds, chemical technology.

SUBSTANCE: invention relates to the improved method for preparing biologically active substances from products of chemical processing vegetable biomass, in particular, to a method for preparing betulinic acid from betuline. Method is carried out by oxidation of betuline with chrome anhydride in acetic acid to betulonic acid, and reduction of betulonic acid with sodium boron hydride is carried out in diethyl ether solution being without its preliminary isolation. Invention provides enhanced yield, simplifying technology, decrease of the process time and decreasing set of solvents used.

EFFECT: improved preparing method.

1 cl, 1 ex

 

The invention relates to methods of producing biologically active substances from the products of chemical processing of plant biomass, in particular to a process for the preparation of betulin Betulinol acid formula

Betulin acid (3β-hydroxy-20(29)-lupen-28-OIC acid) is a natural bioactive substance, the overwhelming growth of melanoma and other cancer cells. Betulin acid found in the bark and leaves of many plants, but its allocation is little viable because of the low content (about 0.1% of dry weight).

There are several ways to get Betulinol acid from betulin content in bark reaches 35%. Chetyrehstoronny way to obtain Betulinol acid from betulin is blocking both hydroxyl groups by acetylation, selective hydrolysis of the ester group of the primary alcohol oxidation released hydroxymethylene groups to carboxyl and final removal of the acetyl protective group (Rucicka L., Lamberton, A.N., Christle C.W. // Helv. Chim. Acta. 1938. V.21. P.1706-1717).

The disadvantage of this method is the low yield of the target product (about 30%), multi-stage, low-tech, the duration of the process.

Known two-stage method of obtaining Betulinol acid (Darrick S.H.L. Kim, Z. Chen Nguyen V.T., Pezzuto J.M., Qiu, S., Lu, Z.Z. // Syn. Comm. 1997. V.27, No. 9. P.1607-1612), in which betulin oxidized with Jones reagent in betulonovoi acid, and then restore the sodium borohydride in tetrahydrofuran for 12 hours at 0°C. the Decomposition of the excess sodium borohydride spend diluted hydrochloric acid. The allocation of Betulinol acid is carried out by distillation, 50% of tetrahydrofuran under vacuum, followed by dilution of the reaction mixture with ethyl acetate, washing the organic solution, removal of the solvent under vacuum. Betulinol acid is obtained in the form of a mixture of 3α and 3β gidroxiizomera in the ratio 1:19.

The disadvantages of this method are insufficient output Betulinol acid (50% from the original betulin) and the duration of the whole process, especially the first stage separation and purification betulone acid. Due to the low solubility betulone acid and sodium borohydride during the process of recovery requires a significant amount of tetrahydrofuran, also uses the second solvent is ethyl acetate.

The closest technological entity and the result achieved by the present invention is a two-stage method of obtaining Betulinol acid, comprising the oxidation of betulin to betulone acid with chromic anhydride in acetic acid at temperature the round 15-20° C for 10 minutes, the extraction betulone acid diethyl ether, washing the ether solution of 10% NaCl, translation free betulone acid into its sodium salt when adding to the ethereal solution of 4 N NaOH solution, separation by filtration of the obtained sodium salt betulone acid, its drying in vacuum in the presence of phosphorus pentoxide, subsequent cleaning of the sodium salt betulone acid from traces of chromium compounds by dissolving it in methyl alcohol and passing the resulting solution through a layer of silica gel, followed by evaporation of the solution and obtaining the sodium salt betulone acid (yield 59%), translation betulone acid in the free state of decay sodium salt betulone acid 10%hydrochloric acid, extraction of the free betulone acid diethyl ether, washing the ether solution with water, separation of the ether solution, drying over sodium sulfate, evaporation and recrystallization from methyl alcohol. The second stage involves the restoration betulone acid to Betulinol in tetrahydrofuran with sodium borohydride at a temperature of 0°C for 10 hours, the decomposition of excess sodium borohydride solution of 2N hydrochloric acid and concentrating the solution by distillation under vacuum of 50% tetrahydrofuran, dilute the concentrate produced is by ethyl acetate, rinse it with water and 10% NaCl, Department of an ethyl acetate solution of Betulinol acid and drying over magnesium sulfate, distillation of the ethyl acetate under vacuum, the recrystallization of the resulting product from methanol. The output of Betulinol acid is less than 50% in terms of the initial betulin (Le Bang Sean, Kaplun A.P., ALEXANDER Shpilevsky, Andía-True YU.S., Alekseeva YEAR, Grigoriev B.B., Shvets V.I. // Bioorganic chemistry. 1998. 24. No. 10. S-793).

The disadvantage of this method is the low output (less than 50%), and the complexity and duration of the process, as each stage includes a large set of different types of processing (washing, cleaning, drying, separation, isolation, repeated distillation of the solvents and the like). Recovering the solvent used is expensive and scarce tetrahydrofuran.

The objective of the invention is to increase the yield of the target product, simplifying technology, shortening the duration of the process and to reduce the set of used solvents.

This object is achieved in that in the method of obtaining Betulinol acid, comprising the oxidation of betulin oxide chromium (VI) in acetic acid in Betulinol acid purification through sodium salt with subsequent transfer to the free betulonovoi acid, its extraction in diethyl ether is recovering betulone acid with sodium borohydride to Betulinol acid, according to the invention restoring betulone acid to Betulinol carried out using a solution betulone acid in diethyl ether, obtained after translation of the sodium salt betulone acid in free betulonovoi acid.

Comparative analysis with the prototype shows that the distinctive features of the prototype feature is the restoration of the sodium borohydride in the solution of diethyl ether, obtained after translation of the sodium salt betulone acid in free betulonovoi acid, without its separation from the solution, i.e. the second stage is carried out in the same solvent used in the first stage of the process. By conducting recovery betulone acid with sodium borohydride in the solution of diethyl ether were able to increase the output of Betulinol acid up to 65% of the original betulin (prototype - 50%), as well as to decrease the number of used solvents (prototype - diethyl ether, methyl alcohol and ethyl acetate). In addition, in the present invention were able to reduce several operations, so when cleaning betulone acid chromium compounds in the prototype, the sodium salt is dried in the presence of phosphorus pentoxide, dissolved in methyl alcohol and passed through silica gel and then evaporated solution. In the proposed method, these operations are simplified.

After recovery of the sodium salt of the free betulonovoi acid is not isolated from the solution, i.e. reduced operation evaporation and recrystallization from methyl alcohol. By reducing these operations were able to simplify the process and shorten its duration.

An example of a specific implementation.

In flask 2 l load 15 g of betulin, add 600 ml of glacial acetic acid. Then to the mixture in one portion was added a solution containing 15 g of chromic anhydride in 20 ml of acetic acid and 22 ml of water, with stirring and at a temperature of 15-20°C. all betulin is dissolved. After 10 min the reaction mixture was diluted with 600 ml of 10% aqueous solution of NaCl. The reaction mass is extracted in a separating funnel diethyl ether 2 times in 600 ml. of Ether extract is washed 4-6 times with 500 ml of 10% aqueous solution of NaCl. In a separating funnel with a volume of 2 l to ethereal extract was added 500 ml of 10% NaCl solution and 300 ml of 4 N NaOH solution at the interface of water and the ether layer is formed flocculent precipitate sodium salt betulone acid. The aqueous and ether layers merge from a separating funnel, and the remaining sodium salt betulone acid washed 2-3 times in a separating funnel 500-600 ml of diethyl ether, then to the sodium salt betulone acid add 1000 ml of diethyl ether and 200 ml of 10% solution is hydrochloric acid, shaken to dissolve betulone acid. The ether layer containing betulonovoi acid, is separated, washed with water, dried over sodium sulfate.

The second stage of recovery betulone acid. The solution betulone acid in diethyl ether (volume of about 1000 ml)obtained after translation of the sodium salt betulone acid in free betulonovoi acid, is transferred into a three-neck 2 l flask equipped with stirrer and reflux condenser, then gradually in the course of 2 hours with vigorous stirring contribute in the reaction mixture of 8 g of sodium borohydride at a temperature of 15-20°C and stirred for 8 hours. The process of recovery betulone acid control by TLC on Silufol plates. Then add 20 or 30 ml of 10% hydrochloric acid. A solution of Betulinol acid in diethyl ether was washed with 400 ml of 10% NaCl solution and 400 ml of water. The ether layer is separated, dried over sodium sulfate. After removal of the ether are Betulinol acid with a yield of 65%. After recrystallization from methanol receive white needle crystals of Betulinol acid TPL296-298°C.

Thus, the proposed method for Betulinol acid can improve the yield of the target product to 65% in relation to the original betulina, as well as to simplify the process, excluding a few article is Dios. In addition, the method is only used diethyl ether, i.e. reduced set of used solvents.

The method of obtaining Betulinol acid,

including the oxidation of betulin oxide chromium (VI) in acetic acid in betulonovoi acid purification through sodium salt with subsequent transfer to the free betulonovoi acid, its extraction in diethyl ether and recovery betulone acid with sodium borohydride to Betulinol acid, characterized in that the recovery betulone acid to Betulinol carried out using a solution betulone acid in diethyl ether, obtained after translation of the sodium salt betulone acid in free betulonovoi acid.



 

Same patents:

FIELD: medicine; pharmaceutical industry; perfumery-cosmetic industry; methods production of betulin.

SUBSTANCE: the invention is pertaining to the method of extraction of betulin from the wastes products of wood-processing, in particular, a birch bark processing and may be used in medicine, pharmaceutical and perfumery-cosmetic industries. The method provides for activation of a birch bark by the shock-acoustic impulses, an alkaline hydrolysis and an extraction of betulin by an alcohol. The operations are conducted simultaneously. The technical result of the invention is simplification of the method of the process, reduction of its duration and power input.

EFFECT: the invention ensures simplification of the method of the process, reduction of its duration and power input.

1 cl, 2 ex

FIELD: chemical technology.

SUBSTANCE: invention relates to methods for preparing oleanolic acid used as a standard sample (comparison samples) in carrying out standardization of medicinal vegetable raw and phytopreparations comprising triterpene saponins - derivatives of oleanolic acid. For preparing oleanolic acid method involves sum of saponins extracted by alkaline extraction from sugar or table beet root crops followed by reprecipitation in acid medium and extraction of precipitate with ethanol, chloroform. Method provides preparing oleanolic acid of high purity degree from inexpensive and available raw.

EFFECT: improved preparing method.

2 cl, 3 ex

FIELD: chemical technology, natural materials, medicine, pharmacy.

SUBSTANCE: invention relates to the improved method for preparing betulin from betulinic acid that can be used in preparing anti-tumor and anti-HIV medicinal preparations. Method for preparing betulinic acid involves oxidation of betulin with chrome (VI) oxide in acetic acid to betulonic acid and reduction with sodium boron hydride to betulinic acid. Betulonic acid sodium salt is reduced to betulinic acid and reduction reaction is carried out at room temperature at the concentration of sodium boron hydride 1.0-6.0 wt.-%. Invention provides simplifying method for preparing betulinic acid, reducing its cost and enhancing ecological safety of the process of it producing.

EFFECT: improved preparing method.

1 cl, 4 ex

The invention relates to new biologically active compounds, namely diglyceride glycyrrhizic acid methyl ester L-valine of formula (I), stimulating primary immune response

The invention relates to new chemical substance, specifically to biologically active compounds having immunostimulatory and antiviral activity (anti-HIV and anti-herpes), - N'-{N-[3-oxo-20(29)-lupen-28-oil] -9-aminopentanoic} -3-amino-3-phenylpropionic acid of the formula I

The invention relates to methods of isolating biologically active substances from waste wood, namely the allocation method Betulinol of the outer layer of birch bark (bark)

The invention relates to new chemical compounds, in particular saponin glycyrrhetic acid - 6',6"-di--D-galactopyranosyl 3-O-[2'-O-(-D - glucuronidase)--D-glucuronidase) methyl ester of glycyrrhetic acid of formula IIB, showing antiulcer activity

The invention relates to new chemical compound, namely the amide glycyrrhizic acid with 5-aminouracil formula I exhibiting anti-HIV activity in cell culture MT-4, with high efficiency by inhibiting the accumulation of virousspecificakih protein P24 (ID50= 55 ág/ml or 52,8 μm), the total viral antigen (ID50= 75 μg/ml or 72,0 μm), which reduces the activity of the reverse transcriptase (reverts) (RT) (ID50= 55 ág/ml or 52,8 μm)

The invention relates to new chemical compounds, specifically to glycopeptide glycyrrhizic acid S-benzyl-L-cysteine [I]: 3-O-{2-O-[N-(-D-glucopyranosyloxy)-L-cysteine(S-benzyl)-N-(-D-glucopyranosyloxy)-L-cysteine(S-benzyl)]}-(3,20)-11-oxo-Olean-12-EN-30-(N-carbonyl-L-cysteine(S-benzyl)-3-yl of the formula (IB), exhibiting anti-HIV activity

FIELD: medicine; pharmaceutical industry; perfumery-cosmetic industry; methods production of betulin.

SUBSTANCE: the invention is pertaining to the method of extraction of betulin from the wastes products of wood-processing, in particular, a birch bark processing and may be used in medicine, pharmaceutical and perfumery-cosmetic industries. The method provides for activation of a birch bark by the shock-acoustic impulses, an alkaline hydrolysis and an extraction of betulin by an alcohol. The operations are conducted simultaneously. The technical result of the invention is simplification of the method of the process, reduction of its duration and power input.

EFFECT: the invention ensures simplification of the method of the process, reduction of its duration and power input.

1 cl, 2 ex

FIELD: organic chemistry, steroids, chemical technology.

SUBSTANCE: invention describes a method for preparing 3-keto-7α-alkoxycarbonyl-substituted ▵4,5-steroid of the formula (I): wherein is taken among or R3 means hydrogen atom (H), lower alkyl, lower alkoxy-group or cyano-group (CN); R21 means hydrogen atom (H) or alkyl; R26 means (C1-C4)-alkyl; R8 and R9 form in common heterocyclic ring system. Method involves interaction of an alkylating agent with 4,5-dihydro-5,7-lactone steroid of the formula (II): wherein R18 means (C1-C4)-alkyl or R18O-group taken in common form O,O-oxyalkylene bridge or keto-group and R3, R8 and R9 have above given values in the presence of a base. Compounds of the formula (I) are used as intermediate compounds in improved methods for synthesis of epoxymexerone.

EFFECT: improved preparing method.

56 cl, 42 tbl, 30 sch, 5 dwg, 89 ex

FIELD: chemical technology.

SUBSTANCE: invention relates to methods for preparing oleanolic acid used as a standard sample (comparison samples) in carrying out standardization of medicinal vegetable raw and phytopreparations comprising triterpene saponins - derivatives of oleanolic acid. For preparing oleanolic acid method involves sum of saponins extracted by alkaline extraction from sugar or table beet root crops followed by reprecipitation in acid medium and extraction of precipitate with ethanol, chloroform. Method provides preparing oleanolic acid of high purity degree from inexpensive and available raw.

EFFECT: improved preparing method.

2 cl, 3 ex

FIELD: organic chemistry, steroids, medicine, pharmacy.

SUBSTANCE: invention relates to steroid compounds of the formula (1)

wherein --- means optional double bonds; R6 means hydrogen atom (H), =CH, -CH3 or -CH2-CH3; R7 means hydrogen atom (H), (C1-C4)-alkyl, (C2-C5)-alkenyl, or (C2-C5)-alkynyl; R11 means hydrogen atom (H), (C1-C4)-alkyl, (C2-C4)-alkenyl, (C2-C4)-alkynyl, (C1-C4)-alkylidene; E means 5-7-memberd ring formed with 16 and 17 carbon atoms at α,cis-position relatively to steroid structure and comprising possibly up to two double bonds. Compounds can be used in therapy and in methods for selective modification of activity of estrogen receptors.

EFFECT: improved method for modifying, valuable medicinal properties of compounds.

10 cl, 1 sch, 1 tbl, 1 ex

FIELD: chemical technology, natural materials, medicine, pharmacy.

SUBSTANCE: invention relates to the improved method for preparing betulin from betulinic acid that can be used in preparing anti-tumor and anti-HIV medicinal preparations. Method for preparing betulinic acid involves oxidation of betulin with chrome (VI) oxide in acetic acid to betulonic acid and reduction with sodium boron hydride to betulinic acid. Betulonic acid sodium salt is reduced to betulinic acid and reduction reaction is carried out at room temperature at the concentration of sodium boron hydride 1.0-6.0 wt.-%. Invention provides simplifying method for preparing betulinic acid, reducing its cost and enhancing ecological safety of the process of it producing.

EFFECT: improved preparing method.

1 cl, 4 ex

FIELD: organic chemistry, steroids, pharmacy.

SUBSTANCE: invention describes unsaturated 14,15-cyclopropanoandrostanes of the general formula (I):

wherein R1 means hydrogen atom (H), hydroxy-group (OH); R2 means hydroxy-group (OH), hydrogen atom (H); R3 means hydrogen atom (H), (C1-C10)-alkyl at α- or β-position; R4 means halogen atom (F, Cl, Br) or pseudohalogen group (azide, rhodanide), hydroxy-group (OH), perfluoroalkyl; R5 means (C1-C4)-alkyl; if double bond is at 1,2-position then R4 can mean hydrogen atom (H). Also, invention relates to a method for preparing these compounds and pharmaceutical compositions containing these compounds. Compounds of the formula (I) are compounds eliciting gestagenic and/or androgenic effect.

EFFECT: improved preparing method, valuable medicinal properties of compounds.

11 cl, 1 tbl, 9 ex

The invention relates to steroids of General formula I

where R1- Oh, (H,H), (H,OR), NOR, where R is H1-6alkyl, C1-6acyl; R2- H or C1-6alkyl, R3- H, or R3- C1-6alkyl, C2-6alkenyl,2-6quinil, possibly substituted with halogen, R4- H, C1-6alkyl or C2-6alkenyl; R5- C1-6alkyl, R5- H, R7- H, C1-6alkyl, R8Is H, OH, halogen;

R9and R10independently H, or R9and R10independently C1-6alkyl, possibly substituted C1-4alkoxy or halogen;

R11- H, SO3H1-15acyl, dashed line indicates a possible link from4,5(10)or4,9-diene system

The invention relates to new chemical substance, specifically to biologically active compounds having immunostimulatory and antiviral activity (anti-HIV and anti-herpes), - N'-{N-[3-oxo-20(29)-lupen-28-oil] -9-aminopentanoic} -3-amino-3-phenylpropionic acid of the formula I

FIELD: organic chemistry, steroids, pharmacy.

SUBSTANCE: invention describes unsaturated 14,15-cyclopropanoandrostanes of the general formula (I):

wherein R1 means hydrogen atom (H), hydroxy-group (OH); R2 means hydroxy-group (OH), hydrogen atom (H); R3 means hydrogen atom (H), (C1-C10)-alkyl at α- or β-position; R4 means halogen atom (F, Cl, Br) or pseudohalogen group (azide, rhodanide), hydroxy-group (OH), perfluoroalkyl; R5 means (C1-C4)-alkyl; if double bond is at 1,2-position then R4 can mean hydrogen atom (H). Also, invention relates to a method for preparing these compounds and pharmaceutical compositions containing these compounds. Compounds of the formula (I) are compounds eliciting gestagenic and/or androgenic effect.

EFFECT: improved preparing method, valuable medicinal properties of compounds.

11 cl, 1 tbl, 9 ex

Up!