Dipolomatic glycyrrhizic acid methyl ester l - valine, stimulating primary immune response
(57) Abstract:
The invention relates to new biologically active compounds, namely diglyceride glycyrrhizic acid methyl ester L-valine of formula (I), stimulating primary immune response.
The inventive compound is moderately hazardous substance and stimulates antibody productive cells in 2 times in comparison with control. table 1.
The invention relates to new biologically active compounds, specifically to diglyceride glycyrrhizic acid methyl ester L-valine of formula (I), stimulating primary immune response.
The specified connection and its properties are not described in literature.
The task, which directed the claimed technical solution is to search for new compounds in the series of derivatives of glycyrrhizic acid (GA), with immunostimulatory activity. In the claimed technical solution synthesized a new molecule - dipolomatic Ledger with methyl ester of L-valine of formula (I), stimulating primary immune response (production of antibody productive cells, UCK).
The method of obtaining the proposed connection (1) Zaid (BCA) in dioxane at 0 - +5 ° C for 3 h with a molar ratio of chemicals VOC: HOSu: BCA=1:4:2 mmol, stand 8-10 hours in the refrigerator at +4 - +8C, the precipitate is filtered off, dichloraniline, and to the filtrate was added to 2.5-2.7 mmol of methyl ester hydrochloride, L-valine and an excess of a tertiary base (triethylamine) (1 ml). The mixture was kept at 20-22 ° C, with periodic stirring for 15-20 h, diluted with cold water, acidified with citric acid to pH 4, the precipitate is filtered off and dried. Purification of the product is carried out by chromatography on silica gel L (40/100 μm, Czech Republic), elwira mixture HCl3-MeOH-H2O, 200:10:1, 100:10:1 (50:10:1) (V%). The output of diglyceride (I) was 60%.
Acute toxicity of diglyceride (I) was determined on outbred mice of both sexes weighing 18-20 g at single intraperitoneal injection. The toxicity parameters were calculated by Litchfield and Wilcoxon signed [Belenky, M. L. elements of a quantitative evaluation of the pharmacological effect (2nd ed., Rev. and supplementary). Leningrad, 1963. S. 152].
Acute toxicity (LD50) was 1100 mg/kg of This compound belongs to the 3rd class moderately hazardous substances [Belenky, M. L. elements of a quantitative evaluation of the pharmacological effect (2nd ed., Rev. and supplementary). Leningrad, 1963. With the method of the NRN and Nordin [Jerne, N. K., Nordin E. A. Plaque formation in by single antibody-producing cells.//Science. 1963. V. 140. P. 405] in the modification Cunningham [Cunningham A. J., Czenberg A. Further improvement in the plaque technique for detecting single antibody forming cell.//Immunology. 1968. No. 2. P. 599]. Animals were immunized by intraperitoneal introduction of a 5% suspension of sheep red blood cells, 24 hours after immunization intraperitoneally injected glikopeptid (I) in a dose of 2 mg/kg On the effect of the studied compounds on humoral immunity was assessed by the number of antibody productive cells (AFC) in the entire spleen, which was estimated visually by the number of zones of hemolysis was then recalculated AOK 106the splenocytes. As the comparison drug used known immunostimulant class of glycopeptides, N-acetylaspartate (TIR) at a dose of 2 mg/kg [L. A. Baltina, G. A. Tolstikov. Muramylpeptide. Ekaterinburg: Ural branch of the Russian Academy of Sciences, 1998. 348 C.]. The results of the experiments presented in the table.
Found that intraperitoneal injection of dipolomatic (I) increases the number of AFC in the spleen in mice at a dose of 2 mg/kg ~ 2-fold compared with control. Dipolomatic (I) stimulate humoral immune response more effective than the reference drug TIR when introduced into equal doses.
The influence of glycopeptide (I) on the production of the KLA in outbred mouse is the cue properties of synthetic glycopeptides-stimulants immunogenesis of nonspecific resistance.//Zhur. microbiol., Epidemiol., immunol. 1981. No. 8. S. 13]. LD50for glycopeptide (I) 1100 (7851540) mg/kg (intraperitoneal, mouse), the connection class III (moderately hazardous substances.
The invention is illustrated by the following examples.
Example 1. The effect of compound (I) on a primary response was studied on 24 outbred white mice weighing 18-20 g according to the method of EPHE and Nordin [Jerne, N. K., Nordin, E. A. Plaque formation in afar by single antibody-producing cells.//Science. 1963. V. 140. P. 405] in the modification Cunningham [Cunningham A. J., Czenberg A. Further improvement in the plaque technique for detecting single antibody forming cell. // Immunology. 1968. No. 2. P. 599]. Animals were immunized 5% suspension of sheep erythrocytes (EB) intraperitoneally for 4 days prior experience. A day after immunization the animals intraperitoneally once entered the compound (I) in a dose of 2 mg/kg Over 4 days (5th) determined the amount of the KLA throughout the spleen. After incubation in a thermostat at t=C for 2.5 h was estimated visually the number of zones of hemolysis, corresponding to the number of the KLA, and recalculated the KLA on 106the splenocytes.
The effect of diglyceride (I) was compared with the effect of MIS on the development of the KLA (see table). at the dose of 2 mg/kg
On outbred mice dipolomatic (I) in a dose of 2 mg/kg increases the amount of AFC on ptx2">Example 2. 3-0-[2-0-N-(-D-glucopyranosyloxy)-L-valine methyl ester]-[N-(-D-glucopyranosyloxy)-L-valine methyl ester]-3,20)-11-oxo-30-norolean-12-EN-3-yl-30-carboxylic acid (I).
To a solution of 0.82 g (1 mmol) glitzirrizinova acid in 50 ml of dioxane at 0 - +5 ° C is added with stirring and 0.46 g (4 mmol) of N-hydroxysuccinimide and stirred at this temperature for 3 h, was kept over night in the refrigerator at +4 to+8C. The precipitate was filtered dicyclohexylmethane and the filtrate cooled to 0 - +5 ° C, was added 0.45 g (2.7 mmol) of methyl ester hydrochloride, L-valine and 0.5 ml (4.9 mmol) of triethylamine.
The mixture was stirred at 20 to 22 ° C for 20 h, diluted with cold water, acidified with citric acid to pH ~4, the precipitate was filtered and washed with water. Dry residue (1.0 g) was chromatographically on a column of silica gel L (40/100 μm), elwira mixture SNS3-Meon-N2ABOUT=200:0:1, 100:10:1, 50:10:1 (V%). Yield 60% (white powder). []20D+ 32 (0,04; Meon). IR spectrum , cm-1: 3600-3200 (HE, NH); 1740 (Sooma); 1660 (CII=0); 154=(CONH). UV spectrummaxMeon (lg ):250 nm (4,95). Found, %: N 2,72 C54H84N2O18. Calculated, %: N 2,67. An NMR spectrum13(CD3OD, 25 ° C, 75.5 MHz, 8, M. D.): 40,5 (C1); 90,6 (C3); 56,7 (C5); 34,1 (C7); 63,4 (C9); 202,9 (C11); to 129.2 (C12); 171,7 (C13); 46,8 (C14); 28,7 (C16)4’); 78,0 (C5’); 171,7 (C6’); 105,2 (C1”); 76,1 (C2”); 76,5 (C3”); 73,6 (C4); 77,3 (C5”); 171,7 (C6”). Fragments of amino acids: C1:53,04; 52,93; C2: 173,36; 173,17; C3: 59,29; 58,70; C4: worth 32.55; C5: 19,73; 19,61.
Thus, the claimed compound (1) is moderately hazardous substance that stimulates the KLA in 2 times in comparison with control. Immunostimulating effect of glycopeptide (I) is more pronounced than TIR.
Dipolomatic glycyrrhizic acid methyl ester L-valine General formula (I)
stimulate a primary immune response.
-D-galactopyranosyl 3-O-[2'-O-(
-D - glucuronidase)--D-glucuronidase) methyl ester of glycyrrhetic acid of formula IIB, showing antiulcer activity
-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
, 20-Olean-12-EN-29-OIC acid of the formula III, R4=H, R5--D-glucuronidase, R6=R7=H and R8= C(O)OH, or, if n= 7, R3=N, R4and R7- simple connection, R5and R6= H or (CH2CH(CH3)O)mH, where m=1 to 14, and R8=CH2OH or CH2O(CH2CH(CH3)O)mH, where m=1-14, generating nitric oxide and activating the soluble form of guanylate cyclase (RGC), antispasmodic, vasodilator and hypotensive means quick action and platelet aggregation inhibitors, method for their production and pharmaceutical compositions based on
means simple or double bond, provided that when
- double bond, R
< / BR>showing hepatoprotective and anti-HIV activity
where R is hydrogen, (C1-C6)alkyl, and the alkyl group optionally contains one phenyl substituent, which, in turn, optionally contains at least one Deputy, selected from the group comprising halogen, methoxy, ethoxy, (C1-C6)alkyl; R1means phenyl cycle containing at least one Deputy, selected from the group comprising (C1-C6)alkoxy, hydroxy, nitro, (C1-C6)alkoxycarbonyl one or fluorine, or R1represents the balance of the pyridine of the formula II
where the carbon atoms 2, 3 and 4 of the remaining pyridine optionally have the same or different substituents R5and R6and R5and R6denote (C1-C6)alkyl or halogen, or R1presents arylamination-2-methylprop-1-ilen group, or R and R1together with the nitrogen atom to which IGN="ABSMIDDLE">
where R7denotes phenyl or pyridinyl; R2means (C1-C6)alkyl, which optionally contains a phenyl residue, which, in turn, optionally substituted with halogen, methoxy group or ethoxypropane, or related to R2(C1-C6)alkyl group optionally substituted 2-, 3 - or 4-pyridinium residue; R3and R4are the same or different substituents and represent hydrogen, hydroxy, (C1-C6)alkoxy, (C1-C3)alkoxycarbonyl or (C1-C3)alkoxycarbonyl(C1-C3)alkyl, or R3is cyclopentanecarbonitrile; Z denotes Oh, and alkyl, alkoxy or alkylamino mean as an unbranched group, such as methyl, ethyl, n-propyl, n-butyl, n-hexyl and branched alkyl groups such as isopropyl or tert-butylene group; halogen means fluorine, chlorine, bromine or iodine and alkoxygroup means methoxy, propoxy, butoxy, isopropoxy, isobutoxy or phenoxypropan, and their pharmaceutically acceptable salts with acids
-oxidase from the fungus Trichoderma and method for producing this enzyme
