H-phosphonates hydroxyl-containing steroids as a reagent for solid-phase synthesis of steroid derivatives of oligonucleotides or their analogs
(57) Abstract:Use: as a reagent for solid-phase synthesis of steroid derivatives of oligonucleotides. The inventive product - H-phosphonate hydroxyl-containing steroids total f-ly 1 , where X is the residue of cholesterol or testosterone. Exit 69 - 89% . table 1. The invention relates to bio-organic chemistry, namely to new chemical compounds - N-hydroxyl-containing phosphonates steroids of General formula I XO--H where X is the residue of the hydroxyl-containing steroid,
testosterone (TS) which can be used as source reagents for solid-phase synthesis of steroid derivatives of oligonucleotides and their analogs.Known just one example of solid-phase synthesis cholesterolaemia oligonucleotides based on the use of 2-(cholesteryloxycarbonyl)ethylamine of formula II  NH(CH2)2NH-O-cholesterol obtained from cholesterolaemia and Ethylenediamine (output - 76% ). Using this reagent produced a number of oligodeoxyribonucleotides with the rest of cholesterol associated with 31-end mezhnukleotidnyh phosphate. Output cholesterinspiegel of the oligonucleotide - 50% .Solid phase synthesis of oligonucleotides and their analogues is currently the most widely used method because of its speed and efficiency. Trials of steroid derivatives of oligonucleotides in cell culture systems have shown them to have promise as a means of suppressing the development of viruses. Therefore a very important task is to increase the output steroid derivatives of oligonucleotides and their analogs for solid-phase synthesis of these derivatives.This task is solved by the fact that, as a reagent for solid-phase synthesis steroidogenesis oligonucleotides and their analogues using new chemical compounds - N-hydroxyl-containing phosphonates steroids of formula 1, which is obtained from the appropriate steroid treatment timidation phosphorus, followed by hydrolysis with water and release by chromatography on silica gel.The synthesized H-phosphonates hydrox is the breaking of the oligonucleotides of deoxyribo-series the RIBO-some and 21-0-Metallbau-lost, pre-obtained solid phase H-phosphonate method. The obtained results confirm the possibility of solid-phase synthesis of steroid derivatives of oligonucleotides using this reagent, the yield of target products is 80-90% . Conducting the reaction in a solid-phase version greatly simplifies the procedure of excess reagents and can be easily automated.P R I m e R 1. Synthesis of H-phosphonates hydroxyl-containing steroids.a) Synthesis of H-phosphonate cholesterol.The flask with argon 1.2 g (17.5 mmol) of imidazole, previously dried over P2O5at 1-2 mm RT. Art. , dissolved in 60 ml of abs. acetonitrile and cooled with ice. Then added with stirring to 0.35 ml of 5.25 mmol) PCl3and 1.5 ml (18 mmol) of triethylamine. After 15 min added dropwise a solution of pre-dried cholesterol (0.5 g, 1.25 mmol) in 60 ml of abs. chloroform for 30 min, then the cooling is removed and continue stirring for another 2 hours Over the course of the reaction is monitored by thin layer chromatography (TLC) in the system chloroform - ethanol (9: 1). Then the reaction mixture is decomposed by adding 15 ml of water and evaporated to dryness. A mixture of rastaa2SO4, evaporated and chromatographic on silica gel (column 150 ml, a concentration gradient of ethanol (0 -> 50% ) in chloroform containing 1% of triethylamine). The fractions containing the product with Rf= = 0,23 (TLC, chloroform - ethanol 6: 4), evaporated to dryness. Yield 0.6 g (85% ).The product is homogeneous according to TLC and gives a positive ninhydrin test.The infrared spectrum, cm-1: 1000, 1060, 1090 (cycle), 1240 (P= O), 1340 (-CH), 1460 (S-CH3), 1600 (C= C), 2400, 2500 (R-OH), 2690 (P-H), 2860, 2950 (CH2-, -CH3WITH-N).31P-NMR,M. D.(CDCl3): 0,86 (J = 618 Hz)
1H-NMR,M. D.(CDCl3): OF 0.82 (S, 3H, 13-CH3), TO 0.88 (S, H, -(CH3)2), OF 0.95 (S, 3H, 20-CH2), is 1.31 (t, N, CH3CH2-N), from 0.8 to 2.40 (m, 3H, cycle), 3,03 (g, 6H, CH3CH2-N) of 3.94 (m, 3H, 10-CH3), 5,27 (Sm, 1H, 3-H), 6,98 (d, 1H, P-H (J = 618 Hz)), 8,35 (S, 1H, N-H).The signals involved in phosphorus-containing part of the molecule in the infrared,31P and1H-NMR spectra correlate well with the literature data  . Signals steroid part of the molecules in the spectra of IR and1H-NMR spectrum is also consistent with reference data  .b) Synthesis of H-phosphonate testosterone.The synthesis are similar to those described above. The yield of 0.49 g (69% ). Rf= 0,29 (TLC, chloroform-ethanol 6: 4)CH2, -CH3).UV-spectrum (ethanol): max241 nm, lg = = 4,18.31P-NMR,M. D.(CDCl3): 3,43 m D. (J = 608 Hz).1H-NMR,M. D.(CDCl3): OF 0.68 (S, 3H, 13-CH3), OF 1.34 (S, 3H, 10-CH3), to 1.21 (t, 9H, CH3-CH2-N), 0,80-2,1 (m, 17H, cycle), 2,94 (g, 6H, CH3-CH2-N), 3,95 (m, 1H, 10-CH3), 5,57 (Sm, 1H, 17-H), of 6.68 (d, 1H, P-H (J = 608 Hz)), 8,11 (S, 1H, N-H).The data of IR, UV and NMR are consistent with literature data [2-3] .P R I m m e R 2. The synthesis of steroid derivatives of oligonucleotides.For the synthesis used by solid state method for the automatic synthesizer "Victoria"  protected oligonucleotides in the form of H-phosphonates, connected on 31-end with a polymeric carrier and having on 51-the end free oxygraph.a) Synthesis of cholesterol derived destimulate
ChSpT(pT)9< / BR>To the dried sample (8 mg, capacity 70 µmol/g at the first nucleotide of the link, i.e., 0.5 mmol), polymer-bound destimulate with H-phosphonate mezhnukleotidnyh links, add 50 ál of abs. pyridine, 25 µl of abs. acetonitrile, 25 µl of 0.1 M solution (25 mmol) of H-phosphonate cholesterol in chloroform and 6 ml (75 mmol) of pivaloate. The mixture was incubated 15 min Emese pyridine - water (98: 2), incubated 30 min and washed with acetone. Then the carrier is treated with 1 ml conc. NH4OH for 1 h, the solution is drained, evaporated to dryness, dissolved in 1 ml of water and applied to a column of ion exchange sorbent. Selected fractions again evaporated, dissolved in water and applied on the column with reversed phase. Conditions of chromatography are shown in table. After separating the fraction containing the target product, evaporated to dryness, dissolved in 100 ál of water and added dropwise to 1 ml of 2% aqueous solution of LiClO4in acetone. The precipitation of the lithium salt of the oligonucleotide is separated by centrifugation, washed with acetone and dried under 14-2mm RT. Art. the Output and characteristics of the cholesterol derived destimulate shown in the table.b) Synthesis of testosterone derived destimulate.TSpT(pT)9< / BR>Synthesis and secretion are similar to those described in example 2A. The output and characteristics of the target product are listed in the table.C) Synthesis of cholesterol derived decorrelate.ChSpU(pU)9< / BR>The synthesis are similar to those described in example 2A, the procedure for allocation impose additional step: resulting from the protective groups in the 21-position of the ribose treated with 1 ml of 0.01 M HCl for 2 h at 50aboutC. Then the solution is neutralized conc. NH4OH, evaporated to dryness and dissolved in 1 ml of water. Then put on a column of reversed phase (chromatography conditions shown in the table) and after chromatography produce the target product similar to that described in example 2A. The output and characteristics of the product are shown in table.g) Synthesis of testosterone derived decorrelate.TSpU(pU)9< / BR>Synthesis and secretion are similar to those described in example 2B. The results are shown in the table.d) Synthesis of cholesterol derived DECA(21-0-methyluridine)
ChSpUm(pUm)9< / BR>Synthesis and secretion are similar to those described in example 2A. The results are shown in the table.e) Synthesis of testosterone derived DECA(21-0-methyluridine)
TSpUm(pUm)9< / BR>Synthesis and secretion are similar to those described in example 2A.The results are shown in the table. (56) Letsinger, R. L. , Zhang, G. , Sun, D. K. , Ikeuchi , T., P. S. Sarin PNAS, 1989, v. 86, N 17, p. 6553-6556.P. J. Garegg , Regberg , T., Stawinski, J. Chem. Ser. 1986, v. 26, N 1, p. 59-62.The Sadtler Standard Spectra. Philadelphia Sadtler research Lab. 1966-1988.Veniaminova A. G. , Levin Aeriosa steroids of General formula
where X is the residue of cholesterol or testosterone,
as a reagent for solid-phase synthesis of steroid derivatives of oligonucleotides or their analogs.
FIELD: organic chemistry, steroids, medicine, pharmacy.
SUBSTANCE: invention relates to 3-methylene-steroid derivative of the general formula (1):
wherein R1 means hydrogen atom (H), or in common with R3 it forms β-epoxide; or R1 is absent in the presence of 5-10-double bond; R2 means (C1-C5)-alkyl; R3 means βH, βCH3 or in common with R1 it forms β-epoxide; either R3 is absent in the presence of 5-10-double bond; R4 means hydrogen atom, lower alkyl; Y represents [H, H], [OH, H], [OH, (C2-C5)-alkenyl], [OH, (C2-C5)-alkynyl] or (C1-C6)-alkylidene, or =NOR5 wherein R5 means hydrogen atom (H), lower alkyl; dotted lines represent optional double bond. Compound can relate also to its prodrug used for treatment of arthritis and/or autoimmune diseases.
EFFECT: valuable medicinal properties of compounds, improved method for treatment.
38 cl, 1 tbl, 18 ex
FIELD: organic chemistry, medicine, pharmacy.
SUBSTANCE: invention represents new derivatives of 17,20-dihydrofusidic acid of the formula (Ia)
wherein Q1 and Q2 are similar or different and mean -CO-, -CHOH-, -CHRO- wherein R means (C1-C4)-alkyl; Q3 means -CH2-; Y means hydrogen atom (H); A means -O- or -S-; R1 means (C1-C4)-alkyl, (C2-C4)-olefin, (C1-C6)-acyl, (C3-C7)-cycloalkylcarbonyl, benzoyl. These derivatives are used in pharmaceutical compositions for treatment of infectious diseases, in particular, in composition for topical applying for treatment of infectious diseases of skin and eyes.
EFFECT: valuable medicinal properties of compounds.
22 cl, 7 tbl, 41 ex
FIELD: organic chemistry, chemistry of steroids.
SUBSTANCE: invention relates to a new method for synthesis of 6β-formyl-B-norcholestane-3β,5β-diol of the formula (I): by constricting six-membered B-ring of cholesterol. Method involves photooxidation of cholesterol with air oxygen at irradiation by visible light in the presence of porphyrine photosensibilizing agent immobilized on low-molecular fraction of copolymer of tetrafluoroethylene and perfluoro-3,6-dioxo-5-methyl-6-sulfonylfluoride octene-1 in the mass ratio porphyrine photosensibilizing agent : cholesterol = 1:(12-15). As porphyrine photosensibilizing agent 5,10,15,20-tetraphenylporphyrine can be used. Method shows technological simplicity, it doesn't require rigid conditions and provides the high yield of the end product.
EFFECT: improved preparing method.
2 cl, 3 ex
FIELD: medicinal industry, sterols.
SUBSTANCE: invention relates, in particular, to the improved method for producing sterols - lanosterol and cholesterol from wooly fat that can be used in preparing medicinal and cosmetic preparations. Method is carried out by alkaline hydrolysis of raw, extraction of unsaponifiable substances, removal of solvent and successive isolation of lanosterol and cholesterol. Alkaline hydrolysis of raw is carried out with a mixture of ethanol, sodium hydroxide, pyrogallol and water at temperature 70°C for 4 h at stirring in the following ratio of components: raw : ethanol : sodium hydroxide : pyrogallol : water = 100.0:(300.0-350.0):(30.0-35.0):(0.01-0.05):(7.5-12.0), respectively, with the indicated mixture with addition of toluene in the following ratio: raw : ethanol : sodium hydroxide : pyrogallol : toluene : water = 100.0:(220.0-255.0):(30.0-38.0):(0.05-0.12):(100.0-137.0):(2.5-7.0), respectively, and lanosterol is isolated by precipitation from mixture of methylene chloride and ethanol in the ratio = 1:1. Before removal of solvent unsaponifiable substances are extracted at temperature 50°C for 2-3 h at stirring. Invention provides increasing yield of the end product, enhancing qualitative indices and reducing cost of production.
EFFECT: improved producing method.
2 cl, 3 ex
SUBSTANCE: polyaminosteroid branched derivatives of general formula I are described, where R1 is saturated or unsaturated C2-C10alkyl (conjugated or branched) or methyl, R2 is COOH or branched polyamine fragments, R3 is H, OR19, where R19 is H or C1-6acyl, R4 is H, R5 is H, CH3, R6 is H, CH3, R7=R8=R9=H, R10 is H, CH3, R11 is OH,-OSO3, - O-acyl, -(Z)n-(NR-Z)p-N(R)2, Z is linear hydrocarbon diradical, n=0, 1, p=1, R-H, C1-6alkyl, C1-6aminoalkyl, possibly substituted by C1-6alkyl, R12=R13=R15=H, R16 is H, OH, R17 is H, R18 is H, CH3, possible double bond. Compounds possess bactericidal activity and can be used for prevention of bacterial infections.
EFFECT: production of polyaminosteroid derivatives, possessing bactericidal activity which can be used for prevention of bacterial infections.
27 cl, 31 ex, 1 tbl, 2 dwg
SUBSTANCE: claimed invention describes paroxetine cholate or salt of cholic acid derivative and composition, which contains paroxetine and cholic acid or its derivative. Also described is pharmaceutical composition for treatment of depressive states, containing paroxetine salt or composition. Pharmaceutical composition can be part of peroral medication, swallowed without water, on form of disintegrating in mouth paroxetine tablet.
EFFECT: obtaining paroxetine cholate or salt of cholic acid derivative, which can be used in pharmacology.
19 cl, 38 ex, 12 tbl
SUBSTANCE: invention refers to synthesis of biologically active substances, in particular specifically, to improved method of producing 2,3-monoacetonide 20-hydroxyecdysone of formula found in very small amounts in some plants, e.g. Rhaponticum carthamoides. Method is implemented by interaction of 20- hydroxyecdysone (1.0 g, 2.08 mmole) and acetone with phosphomolybdic acid (PMA) added. As suspension is effected by mother compound in PMA acetone (0.3 g, 0.16 mmole), after 5 min homogenisation of reaction mixture is observed to be steamed by neutralisation with 0.1% sodium hydrocarbonate solution with following ethyl acetate and chromatography extraction of the end product, thus resulting in isolation of the end 2,3-monoacetonide 20- hydroxyecdysone of 32% yield.
EFFECT: method is highly selective and single-stage.
2 cl, 1 ex
SUBSTANCE: claimed invention relates to novel fusidic acid derivatives of general formula [I], where X represents halogen, trifluoromethyl, C1-C7alkyl, substituted with phenyl, C2-C9alkenyl, optionally substituted with C1-C7alkyl, halogen or phenyl, phenyl, optionally substituted with one or two similar or different substituents, selected from group consisting of halogen, C1-C7alkyl, C2-C9alkenyl, phenyl, C1-C6alkoxy, nitro, C1-C6alkyltio, trifluoromethyl and cyano; or X represents naphtyl; Y and Z both represent hydrogen or together with bond C-17/C-20 form double bond between C-17 and C-20 or together represent methylene and form cyclopropane ring in combination with C-17 and C-20; A represents O, S or S(O); B represents C1-6alkyl, C2-6alkenyl, C1-6acyl, phenyl or benzoyl, where C1-6alkyl is optionally substituted with one or more halogens, hydroxy, C2-6alkenyl, phenyl, C1-4heteroaryl or C1-6alkoxy; Q1 represents -(CHOH)-, or -(CHW)-, where W represents halogen or azido; Q2 represents -(CHOH)-; to their pharmaceutically acceptable salts and easily hydrolysed esters and to pharmaceutical compositions, including said derivatives, as well as to their application in therapy.
EFFECT: application in therapy.
31 cl, 127 ex, 5 tbl
FIELD: production processes.
SUBSTANCE: invention refers to wood working and wood chemical industries. Birch bark is broken down, mixed with liquid, the mixture is held at temperature higher than mixture freezing temperature, then triterpene compounds are separated from lingo-adipic residue with the following filtration and drying. Birch bark is additionally broken down by method of impact-abrasing and/or abrasing effect till obtaining birch bark flour. Birch bark flour is mixed with liquid with density of 0.999-0.958 kg/m3. Mixture is held for 0.1-10 hours and then separated by flotation to hydrophobic and hydrophilous fraction. Solution remaining after separation is condensed and dried. Obtained hydrophobic fraction - mixture of triterpene compounds - is exposed to recrystallisation in ethanol with activated charcoal and then betuline, solution of triterpene compounds in ethanol and mixture of triterpene and polyphenol compounds at carbon matrix is obtained. Or triterpene compounds mixture is separated to fractions in carbon-dioxide extractor and betuline, dry mixture of triterpene and polyphenol compounds are obtained. Hydrophilous fraction - lingo-adipic flour - is separated from liquid and dried out.
EFFECT: increase of environmental safety and method effectiveness.
6 cl, 4 ex, 3 dwg
SUBSTANCE: present invention presents a preparation to reduce insulin resistance. The preparation contains 3-O-v-D-glucopyranosyl-4-methylergost-7-ene-3-ole, or an extract made with using an organic solvent, or an extract made with using hot water, or a drained liquid of a plant of Liliaceae family, or fraction thereof which contains this compound as an active component.
EFFECT: production of the preparation which is suitable for inhibition of adipocytokine production, particularly adipocytokine which cause insulin resistance, and for prevention of pathological conditions caused by insulin resistance, or simplification of clinical course of said pathological conditions.
9 cl, 3 ex