Medicament for suppressing infection and human immune deficiency virus proliferation

FIELD: chemical engineering; pharmaceutical engineering.

SUBSTANCE: method involves applying substances of macrolide group showing high inhibition degree with respect to protein of tyrosine kinase.

EFFECT: enhanced effectiveness in suppressing infection and proliferation of human immune deficiency virus in macrophages.

4 cl, 25 dwg

 

Background of the invention

1. The scope of the invention

The present invention relates to an agent for suppressing the infection and proliferation of human immunodeficiency virus (HIV-1), which infects immunocompetent cells such as macrophages or dendritic cells, and causes damage to the immune system. More specifically, the present invention relates to the use of an agent for suppressing the infection and proliferation of human immunodeficiency virus, the agent that inhibits infection and proliferation of human immunodeficiency virus in macrophages type M-derived human monocytes.

2. Description of the prior art,

More than 40 million people infected with human immunodeficiency virus (HIV-1), and it was found that among these, 5 million people have developed the syndrome of acquired immunodeficiency syndrome (AIDS). AIDS really is a worldwide infectious disease. Highly active antiretroviral therapy (HAART) in patients with HIV-1 appeared in the early 90-ies. In the clinical application was received significant therapeutic effect, such as reducing the number of viruses in the blood plasma of patients with AIDS and installed the selection of CD4.

However, on the other hand, it is disturbing that the virus aimed to macrophages (Mϕ), remains in the reticulo endothelially tissues 1),2)and in patients receiving HAART, there has been the emergence of resistant strain to HAART during the period of time from about six months to several years after HAART. In addition, since the cost of treatment with HAART is high, treatment is mainly assigned to people in developed countries, and people in developing countries may not receive such therapy due to inequality in the economic situation in developed countries and developing countries, where time is more than 80% of patients infected with HIV in the world, and such treatment may not contribute to interrupt the international spread of the virus, which results in multiple combination therapy to suppress the emergence of resistant strains, and also reports about patients who do not receive treatment due to abdominal symptoms and damage to the hematopoietic the system, therefore, pointed out the limitations both in medical and social terms when using HAART.

HIV-1 is the virus that infects immunocompetent cells such as macrophages and dendritic cells, with the subsequent destruction of the immune system. In recent studies it is apparent that the infection and proliferation of HIV-1 in macrophages play an important role in maintaining the infection and the development of the pathogenesis of the ICH-1 3)therefore, it requires the development of new drugs that inhibit the infection and proliferation of HIV-1 in macrophages.

Despite conducting numerous experiments on infection of macrophages by HIV-1, many studies were performed using cell strains macrophages, but the results of experiments using cell strains do not always reflect the function of tissue macrophages in vivo. Akagawa et al. succeeded in differential induction of two types of macrophages, contributing to the proliferation of HIV-1, and macrophages that suppress the proliferation of HIV-1 from monocytes, in addition, they additionally showed that macrophages that suppress proliferation, can serve as a model of human alveolar macrophages, causing it became possible to study the infection, proliferation and the mechanism of suppression of proliferation of HIV-1 in the system, similar to macrophages in vivo4)-9).

In addition, it is known that macrolides are effective in the treatment of panbronchiolitis (DPB) and diseases in the field of otorhinology, and a known mechanism of action, in addition to antibiotic action, in relation to the induction of anti-inflammatory action10). In particular, there is a message in the result of the study, the accumulation of macrolides in the tissues, indicating that macrophages were observed in several with the-thousand times higher accumulation compared with peripheral lymphocytes, therefore, it is believed that the effect of macrolides on macrophages is important.

Based on the above assumptions, the inventors hypothesized that is useful for drug design and development, complementary disadvantages of HAART, suppress the infection and proliferation of HIV-1 in macrophages, and is aimed at the destruction of HIV-1, directed to macrophages from lymphoid reticuloendothelial system, particularly the development of cheap chemotherapeutic drugs from the point of view of therapeutic strategies against AIDS worldwide for the treatment of patients infected with HIV-1.

The inventors have investigated, or have no known macrolide derivative action against the suppression of the infection and proliferation of HIV-1 and suddenly discovered that they have a vast effect on the infection and proliferation of HIV-1 in macrophages and showed that the inhibitory effect on the proliferation manifested by suppression of expression of protein tyrosine kinase Hck in macrophages, which is necessary for growth of the virus and suppress activation RMA, and made the present invention.

The aim of the present invention is the enforcement agent to suppress the infection and proliferation of human immunodeficiency virus suitable for the treatment of patients infected with HIV-1, cheap tuberculosis treatment is autocheckin drug also suitable as an additional drug in the HAART.

Brief description of the invention

The present invention relates to the use of macrolide derivatives to suppress the infection and proliferation of human immunodeficiency virus in macrophages derived from human monocytes. Macrophages derived from human monocytes, macrophages are type M. Suppression of proliferation of the virus is based on the inhibitory effect of macrolides on protein tyrosinekinase Hck and the inhibition of activation MRC macrophages required for proliferation of the virus. Known macrolide derivatives having such an overwhelming effect on proliferation of HIV-1, included in the present invention.

Preferred examples of macrolide derivatives used in this invention are:

Oxacyclohexadecan-2,10-dione,4[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexopyranosyl)oxy]-14-ethyl-7,12,13-trihydroxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy]; 11-(1’-hydroxypropyl)-3-[[2,6-dideoxy-3-C-methyl-α-L-RIBO-hexopyranosyl]oxy]-5-[(3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl)oxy]-2,4,6,8,11,14-HEXAMETHYL-10,13,15-three-ocatillo[9.2.1.1.9.6]pentadecane-1-he; 6,15,16-trioxadecyl[10.2.1.11,4]hexadecan derived erythromycin; 4,1-dioxabicyclo [8.2.1]tridec-12-EN-5-Oh,7-[(2,6-dideoxy-3-C-methyl-α -L-RIBO-hexopyranosyl)oxy]-3-(1,2-dihydroxy-1-methylbutyl)-2,6,8,10,12-pentamethyl-9-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy]; oxazolo-tetradecane-2,10-dione,4-[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexopyranosyl)oxy]-14-ethyl-12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy]; de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de (3’-N-methyl)-3’-N-sulfonyl-8,9-anhydrotetracycline

And 6,9-hemiketal or its salt; de(3’-N-methyl)-[3’-N-(3-hydroxy-1-propyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-N-methyl)-3’-N-(2-acetoxyethyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-N-methyl)-3’-N-cyanomethyl-8,9-anhydrous-pseudoalteromonas And 6,9-hemiketal or its salt; de(3’-N-methyl)-3’-N-(2-foradil)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-ethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’,3’-N,N-diethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-allyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de (3’-N-methyl)-3’,3’-N,N-diallyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-m is Teal)-3’-N-propargyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’,3’-N,N-dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-propyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’,3’-N,N-dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-hexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’,3’-N,N-dihexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-benzyl-8,9-anhydrous-pseudoalteromonas And 6.9-

hemiketal or its salt; bis-de(3’-N-methyl)-3’,3’-N,N-dibenzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-(2-propyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de (3’-N-methyl)-3’,3’-N,N-di-(10-bromo-1-decanol)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)-3’-N-acetyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-dimethylamino)-3’-piperidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-dimethylamino)-3’-pyrrolidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-dimethylamino)-3’-morpholino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-dimethylamino)-3’-[hexahydro-1-(1H)-azepine]8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de (hidroxi)de[12-(1-hydroxypropyl)]-12-hydroxyacyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(12-hydroxy)-de[12-(1-hydroxypropyl)]-12-amino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3’-N-methyl)-de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; de(3-O-cladinose)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt and de(3-O-cladinose)-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt.

In order to understand the present invention, is illustrated

the mechanism of suppression of proliferation of HIV-1 focused on macrophages, macrophage-derived human monocytes.

[I] Experimental materials and methods

(1-1) Obtaining macrophages derived from human monocytes and alveolar macrophages

As previously reported11)human mononuclear cells from peripheral blood (RMS) was isolated from light layer of a blood clot from healthy human volunteers using a micro granules loaded with CD14 antibodies (Miltenyi Biotec, Germany), magnetic system for the separation of cells (MACS) (Miltenyi Biotec, Germany) and lifepart (Nycome Inc., Norway), then monocytes were isolated from RMS positive selection.

Macrophages derived monocytes were obtained by culturing the selected monocytes in the presence of M-CSF (colony stimulating factor is acroview) (10 4U/ml, obtained from Morinaga Milk Co.), granulocytes and colony-stimulating factor granulocyte, GM-CSF (500 u/ml, obtained from Japan, Scheling Plough Co., Osaka, or obtained from R & S Genzyme-TECHNE Inc.) in RPMI1640 medium with addition of 10% fetal calf serum (Nissui Seiyaku Co., Tokyo) for 7-8 days.

Macrophages, obtained by differential induction using M-CSF from human monocytes were identified as macrophages type M (Mϕ type M or M-Mϕ) and macrophages, obtained by differential induction using GM-CSF from human monocytes were identified as macrophages type GM (Mϕ type GM or GM-Mϕ). In addition, alveolar macrophages (alveolar Mϕ or a-Mϕ) was prepared as follows. Extracted cells from the alveolar washing liquid in the washing of the alveoli in healthy volunteers their suspended and adhesively to plastic, then washed neadgezivnye cells and the remaining adhered cells were identified as alveolar Mϕ.

(1-2) Infection of macrophages by HIV-1

Infection of macrophages by HIV-1 was carried out as follows. Viral strains aimed at macrophages, HIV-1JR-FLand HIV-1BALused to infect for 2 h in contact with cells M-MϕGM-Mϕ and a-Mϕ (drove up to 2.5×105/well, Flacon No. 3043: Becton Dickinson Labware, Inc., USA) at a low concentration(titer antigen P24 50 ng/ml, TCID50=3000/ml of the virus to a final concentration of 100 PG/ml), and neadsorbirovanne to macrophages, the virus was cleaned and cultivated adding fresh medium. In the case of a medium with M-Mϕ added M-CSF (104U/ml) and in the case of a medium with GM-Mϕ added GM-CSF (500 u/ml).

In these experiments the amount of virus used to contact the infection, brought up to a level of virus in patients available at the stage of carriage, because the level of virus in lung tissue from patients at the stage of carriage of HIV-1 is several tens of PG/ml - several hundred PG/ml, and the level of virus infection of alveolar Mϕ makes multiple copies of the 104cells12).

(1-3) analysis of the infection and proliferation of HIV-1

The proliferation of the virus was investigated by the number selected in the culture supernatant of viral particles after infection, using a sandwich ELISA with two types of anti-P24 antibodies (Nu24 and V13)). As LTR (long terminal repeat)-gag gene primer used the following JAM62 and JAM65. For group PCR used the following primers JAM63 and JAM64 as an internal primer.

JAM62:

5’-GCTTCAAGTAGTGTGTGCCCGTCTG-3’

JAM65:

5’-AATCGTTCTAGCTCCCTGCTTGCCC-3’

JAM63:

5’-GTGTGACTCTGGTAACTAGAGATCC-3’

JAM64:

5’-CCGCTTAATACTGACGCTCTCGCAC-3’.

(1-4) Analysis of expression of protein tyrosine kinase Hck protein and factor transcripts the and EURβ

The expression of protein tyrosine kinase Hck protein and transcription factor C/HEBβ in macrophages was investigated after solubilization of macrophages in the buffer for sample for SDC-PAGE, electrophoresis in 10% SDS-polyacrylamide gel, transfer of protein from gel to immobilon membrane P (Millipore Inc., USA) and analysis of antibodies against these proteins Western blot testing. Antibodies against protein tyrosine kinase Hck (N-30) and antibodies against C/HEBβ (C-19) were obtained from Santacluse Inc. (USA). The results of the Western blot were detected by reagent Amersham ELC (Amersham International plc, Buckinghamshire, UK). The intensity of the bands was expressed by the indicator PSL (photostimulable luminescence) (A/mm2).

(1-5) Treatment of macrophages antimuslim protein receptor Hck and protein transcription factor C/EVRV

Antisense oligonucleotide probe Hck protein and protein/HEBβ (AS), the corresponding sense oligonucleotide probe (S) and Nesmelova oligonucleotide probe that is not related to transcription and translation (NS)was used as follows.

Phosphorotioate-modified AS-Hck;

5’-TTCATCGACCCCATCCTGGC-3’

Phosphorotioate-modified S-Hck;

5’-GCCAGGATGGGGTCGATGAA-3’

Phosphorotioate-modified NS-Hck;

5’-CCATATTTCCCGCTCGCGTG-3’

Phosphorotioate-modified AS-C/EBPβ;

5’-CAGGCGTTGCATGAACGCGG-3’

Phosphorotioate-modified S-C/EURβ;

5 is-CCGCGTTCATGCAACGCCTG-3’

Phosphorotioate-modified NS-C/EBPβ;

5’-CCAGAGAGGGCCCGTGTGGA-3’.

These probes were dissolved in serum not containing RPMI1640 medium, in which was dissolved lipofectin (Life Technology

Inc., USA) at a concentration of 5 μm at room temperature for 30 min, added with RPMI1640 medium that contained 10% FCS, macrophages (final concentration 2 μm), and incubated at 37°C for 24 h After which cells were washed with medium, was added RPMI1640 medium containing 10% FCS, without oligonucleotides, were additionally cultured for 24 h, and then infected with HIV-1.

(1-6) Analysis of activation RMR and ERK1/2

Activation RMR and ERK1/2 was determined using antibodies against RAC, anti-ERK/2 antibody, anti-tyrosine phosphorylation RAC antibodies and anti-tyrosine phosphorylation ERK1/2 antibodies (New England Biolabs, Inc., USA) and the reaction of phosphorylation of these molecules were determined by Western blot testing.

[2] Results

(2-1) the Proliferative response of HIV-1 strain, directed by macrophages in M-Mϕ and GM-Mϕ

(2-1-1) protein P24 in HIV-1-infected M-Mϕ and GM-Mϕ in the culture supernatant

Cells M-Mϕ and GM-Mϕ were infected with HIV-1 strainJR-FLand strain of HIV-1BALand incubated for 14 days. The amount of protein P24 in the culture supernatant of macrophages was determined in the same periods of time. When Liu is the first of the strains of HIV-1 protein P24 was detected in the culture supernatant of M-Mϕ [see figure 1(a) and (b)].

(2-1-2) Cytopathy in HIV-1-infected M-Mϕ and GM-Mϕ

Morphological changes of cells M-Mϕ and GM-Mϕinfected with a strain of HIV-1JR-FLand strain of HIV-1BALobserved depending on time. Education klosterbraeu cells and fused cells was observed on the 2nd day of incubation only in producing virus M-Mϕ and 4-7 day celebrated the formation of multinuclear giant cells in culture [see figure 2(A)]. On the other hand, morphological changes were not observed in GM-Mϕin which there was no replication of the virus [see figure 2(B)]. These results indicate that the cytopathic effect such as the formation of clusters, cell fusion and the formation of MGC in macrophages during infection with HIV-1 focused on macrophages, can be used as an indicator to determine the proliferation of the virus.

(2-1-3) analysis of the intracellular distribution of the protein P24 in HIV-1-infected M-Mϕ and GM-Mϕ

Cells M-Mϕ and GM-Mϕ were infected with HIV-1 strainJR-FLand strain of HIV-1BALand immunologically stained with antibodies to P24 for 8 days after infection. In M-Mϕ watched the expression of the protein P24 in the MGC and similar cells [see figure 3(A)]. In GM-Mϕ did not observe expression of the protein P24 [see figure 3(B)]. Based on these results, it was found that the mechanism that suppresses output is s virus in GM-Mϕ can be overwhelming mechanism before intracellular stages of education and active replication of viral particles.

(2-1-4) Detection of viral DNA in HIV-1-infected M-Mϕ and GM-Mϕ

Cells M-Mϕ and GM-Mϕ were infected with HIV-1 strainJR-FLand strain of HIV-1BALand detected the formation of viral DNA at day 2 after infection group PCR using primers LTR-dad. The formation of viral DNA were detected in M-Mϕin which the observed proliferation of the virus, and GM-Mϕwhere not observed the formation of viral DNA [see figure 4(a) and (b)]. These results indicated that the infiltration of the virus into cells and the conversion of RNA into DNA occurred even in GM-Mϕ without detection of proliferation of the virus, it shows that the mechanism of suppressing the formation of viruses in GM-Mϕ takes place after the formation of viral DNA.

(2-1-5) Expression of protein tyrosine kinase Hck and protein/HEBβ M Mϕ and GM-Mϕ and changes in the expression under the action of the HIV-1

Since M-Mϕderived from human monocytes, effectively induce the proliferation of HIV-1, and GM-Mϕ inhibit the proliferation of the virus, the inventors studied depend or not differences in susceptibility to infection with HIV-1 from differences in the expression of the protein of the host. The results showed that the expression of protein tyrosine kinase Hck and the actor transcription/HEBβ is different in the two types of macrophages.

Without virus protein tyrosinekinase Hck expressively highly in M-Mϕ and to a lesser extent expressively in GM-Mϕ [see figure 5]. Despite the fact that the expression of protein tyrosine kinase Hck in M-Mϕ increased upon infection with HIV-1 strainBALat 2 days, by contrast, expression of protein tyrosine kinase in GM-Mϕ was reduced [see figure 5]. On the other hand, M-Mϕ when expression of the protein/HEBβ without virus expressively only high-molecular type (37 KD), but in GM-Mϕ was detected expression of both proteins of high molecular weight type and low-type (23 KD) [see Fig.6]. Although it has not been established changes in the expression of the protein/HEBβ M Mϕ at day 2 after infection with HIV-1 strainBALexpression of low molecular weight protein (HEBβ was significantly increased in GM-Mϕand there are changes in the ratio of high-molecular type and low-type ratio (L/S) [see Fig.6].

(2-1-6) study of the susceptibility to infection of human alveolar Mϕ HIV-1 and expression of protein tyrosine kinase and protein/HEBβ

It has been shown that GM-Mϕderived from human monocytes using GM-CSF, similar to the human alveolar macrophages in morphology, expression behavior is chastnogo marker the productivity of the formation of active oxygen species and expression of catalase13),14). The inventors have investigated similar or no susceptibility to contamination of GM-Mϕderived from human monocytes, and human alveolar Mϕ.

After infection with HIV-1 strainBALalveolar Mϕ (a-Mϕidentified the viral DNA PCR group and analyzed the proliferation of the virus protein P24 in the culture supernatant using ELISA. Viral DNA was found in all periods of time after infection [see Fig.7(B)], but after 14 days of incubation was not detected neither protein P24, nor the products of the virus [see Fig.7(A)].

As a result of studying the expression of protein tyrosine kinase Hck and protein/HEBβ alveolar Mϕ Western blotting was established decrease in the expression of protein tyrosine kinase Hck and changes in the expression of protein isoforms WITH/HEBβand it was significantly increased expression of low molecular weight type protein (HEBβ (23 KD), and has been a decrease in the ratio of L/S [see Fig]. Based on these results it can be assumed that the mechanism of suppression of proliferation of the virus in human alveolar Mϕ and GM-Mϕderived from human monocytes, can largely be similar.

(2-1-7) decrease in the expression of protein tyrosine kinase Hck in processed antimic the new oligonucleotide-M-Mϕ to the squirrel tyrosinekinase Hck and the suppression of cell proliferation in HIV-1

Cells M-Mϕ processed antimyeloma oligonucleotide (Hck-AS) to the protein of tyrosinekinase Hck within 24 h, and M-Mϕ incubated for 24 h and determined the expression of protein tyrosine kinase Hck. In the result, it was found that the expression of protein tyrosine kinase Hck was significantly reduced compared with the control group (L) [see figure 9]. However, in M-Mϕprocessed semantic probe (Hck-S) or an unrelated probe (Hck-NS), was found to suppress the expression of protein tyrosine kinase Hck [see figure 9].

These cells M-Mϕpre-treated with different oligonucleotides were infected with HIV-1 strainBALand evaluated the expression of protein tyrosine kinase Hck 2 days after infection, decrease in the expression of protein tyrosine kinase Hck was found only in the group with addition of antisense probe (Hck-AS), and suppression of expression of protein tyrosine kinase Hck was not detected in the M-Mϕprocessed semantic probe (Hck-S) or an unrelated probe (Hck-NS) [see figure 10]. Protein content was determined P24 in the culture supernatant after 4, 7 and 10 days after infection. In cells M-Mϕprocessed antimuslim probe (Hck-AS), in all periods of time was found significant amounts of protein P24, but M-Mϕprocessed with ikovym probe (Hck-S) and an unrelated probe (Hck-NS) it was found time-dependent increase of P24 protein level similar to the control group treated only with lipofectin (L) [see figure 10]. These results suggest that the expression of protein tyrosine kinase in M-Mϕ required for proliferation of HIV-1.

(2-1-8) Expression of C/HEBβ and growth response of HIV-1 in GM-Mϕprocessed antimyeloma oligonucleotide WITH/HEBβ

Cells GM-Mϕ processed antimuslim probe to/HEBβ (C/EBPβ-AS) for 24 h and cultured for another 24 h, and GM-Mϕ two days after infection with HIV-1 strainBALrelatively preserved expression of high molecular weight isoforms (37 KD), but the expression of low molecular weight isoforms (23 KD) was significantly reduced with increase of the ratio L/S [see 11]. In addition, HIV-1 cells GM-Mϕ, pre-treated C/EBPβ-AS induced proliferation of the virus, and in the culture supernatant was detected protein P24 [see Fig]. Contributing to the growth of HIV-1 action and changes in the expression of C/HEBβ not found in groups, add/HEBβ-S and C/EBPβ-NS [see 11 and Fig].

The above results of the experiments clearly indicate that cells M-Mϕ and GM-Mϕderived from human monocytes, have different susceptibility to HIV-1, the direction is pushed to the macrophages, and M-Mϕ significantly enhanced the proliferation of the virus, while on the contrary, GM-Mϕ the proliferation of the virus suppressed. Furthermore, the difference in the nature of proliferation in M-Mϕ and GM-Mϕ coincided with the difference in expression of high molecular weight isoforms and low molecular weight forms of the protein tyrosine kinase Hck protein and transcription factor C/HEBβ in these cells. In the specifically regulated the expression of isoforms of the protein tyrosine kinase Hck and protein/HEBβ using antisense oligonucleotides, the replication of the virus in cells M-Mϕ fully regulated and, on the contrary, the reproduction of the virus can be induced in GM-Mϕ.

Based on these results, it clearly follows that tyrosinekinase is required for the proliferation of HIV-1 in M-Mϕand low-molecular type isoforms WITH/HEBβ plays an important role in the suppression of proliferation of HIV-2 GM-Mϕ. In addition, since the results of the study of susceptibility to HIV-1 in human alveolar Mϕ and expression of protein tyrosine kinase Hck and EURβand because the mechanism of inhibition of growth of HIV-1 in human alveolar Mϕ and GM-Mϕderived human monocytes are identical, the analysis in GM-Mϕ may be highly associated with the analysis of the alveolar Mϕ in vivo. The results of these experiments indicate the, the analysis of the mechanism of proliferation of HIV-1 in M-Mϕderived from human monocytes, suitable as the main experimental system for drug development, with overwhelming effect on the growth of HIV-1.

Based on the above results, it can lead to preferable examples of macrolide derivatives having the property of inhibiting the expression of protein tyrosine kinase in M-Mϕderived from human monocytes, and inhibition of activation RMR, which is required for proliferation of the virus, in the form of the following compounds, and these compounds are industrially available or easily obtained by methods described in the literature.

Oxacyclohexadecan-2,10-dione,4[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexopyranosyl)oxy]-14-ethyl-7,12,13-trihydroxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]hydroxy (Sigma Inc., USA); in the following denoted as EAT.

11-(1’-Hydroxypropyl)-3-[2,6-dideoxy-3-C-methyl-α-L-RIBO-hexopyranosyl]oxy]-5-[(3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl)oxy]-2,4,6,8,11,14-HEXAMETHYL-10,13,15-three-ocatillo [9.2.1.1.9.6]pentadecane-1-he (see P.Kurath et al. Exoperimentia, 27, 362, 1971); in the following designated as EM.

6,15,16-Trioxadecyl[10.2.1.11,4]hexadecan, a derivative of erythromycin or 6,9,12-microeletronics (see R. Kurath et al. Exoperimentia, 27, 362, 1971); in the following designated as EM.

4,13-Dioxabicyclo [8.2.1] tridec-12-EN-5-Oh,7-[(2,6-dideoxy-3-C-methyl-α-L-RIBO-hexopyranosyl)oxy]-3-(1,2-dihydroxy-1-methylbutyl)-2,6,8,10,12-pentamethyl-9-[[3,4,6-trideoxy-3-(dimethylamino)-β-3-D-Xylo-hexopyranosyl]oxy] (see JP-A-7-247299); in the following denoted by as EM.

Oxacyclohexadecan-2,10-dione,4-[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexopyranosyl)oxy]-14-ethyl-12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy] (see S.Morimoto et al. J Antibiotics, 43, 286, 1990 or the product Apin Chemicals Ltd., Britain and the product of Wako Pure Chemicals, Inc. Japan); in the following designated as HIMSELF.

Examples of derivatives of erythromycin are:

de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-N-methyl)-3’-N-sulfonyl-8,9-anhydrous-pseudoalteromonas And G,9-hemiketal or its salt, in the following denoted as EM; de(3’-N-methyl)-[3’-N-(3-hydroxy-1-propyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-N-methyl)-3’-N-(2-acetoxyethyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-N-methyl)-3’-N-cyanomethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in sleduushem designated as EM; de(3’-N-methyl)-3’-N-(2-foradil)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-ethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, subsequent designated as EM; bis-de(3’-N-methyl)-3’,3’-N,N-diethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-allyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de (3’-N-methyl)-3’,3’-N,N-diallyl-8, 9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-propargyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’,3’-N,N-di-propargyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-propyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’,3’-N,N-dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de (3’-N-methyl)-3’-N-hexyl-8,9-anhydrous-pseudoalteromonas And 9-hemiketal or its salt, in the following designated as EM; bis-de(3’-N-methyl)-3’,3’-N,N-dihexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-benzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de (3’-N-methyl)-3’,3’-N,N-dibenzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-(2-propyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’,3’-N,N-di-(10-bromo-1-decanol)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; bis-de(3’-N-methyl)-3’-N-acetyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-dimethylamino)-3’-piperidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-dimethylamino)-3’-pyrrolidino-8,9-the anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de (3’-dimethylamino)-3’-morpholino-8, 9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3’-dimethylamino)-3’-[hexahydro-1(1H)-azepine]8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the PEFC is blowing designated as EM; de(12-hydroxy)-de[12-1-(hydroxypropyl)]-12-hydroxyacyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de[12-(hydroxypropyl)]8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(12-hydroxy)-de[12-(1-hydroxypropyl)]-12-amino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, later identified as EM; de(3’-N-methyl)-de[12-1-(hydroxypropyl)]8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM; de(3-O-cladinose)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt, in the following denoted as EM and de(3-O-cladinose)-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or salt, in the following denoted as EM.

Various of the above derivatives of erythromycin were established by the authors of the present invention Satoshi Omura et al., and filed as an international patent application, international publication WO 02/14338 A1, including pseudoalteromonas, which, as expected, is a new anti-inflammatory agent. In the description of WO 02/14338 A1 detailed methods of synthesis and chemical structure of erythromycin derivatives, and descriptions of methods of synthesis of each derivative of erythromycin are explained below.

Synthesis AM

The solution erythromyc is on (12.4 g) in glacial acetic acid was stirred at room temperature for 2 h, was slowly added an aqueous sodium bicarbonate solution and neutralized. The reaction mixture was extracted with chloroform, the organic layer was obezvozhivani the mirabilite, was filtered mirabilite and the solvent was removed by evaporation to obtain crude solid. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (7.7 g). Then a methanol solution EM (7.6 g) was added potassium carbonate (1.4 g) and boiled under reflux for 2 hours After removal of the solvent the residue was dissolved in aqueous sodium bicarbonate solution and was extracted with chloroform. The mixture was obezvozhivani the mirabilite, filtered and removed the mirabilite, then the resulting crude solid was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.5) with obtaining EM (5.9 g, white powder).

Synthesis of de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Sodium acetate (3.9 g) and iodine (2.5 g) were added in that order to a solution EM (6,9 g) in methanol (52,0 ml) - water (13,0 ml) at room temperature and was stirred at 50°C for 3 hours while stirring was added 1 N. aqueous solution of sodium hydroxide to maintain pH 8-9. After confirming completion of the reaction p is TLC, the reaction mixture was diluted with a mixture of aqueous ammonia (7.5 ml) - water (200 ml) and was extracted with dichloromethane. After dehydration of the organic layer by mirabilite mirabilite was removed by filtration and the solvent drove away with getting wet substances. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (4.8 g, white powder).

Synthesis of bis-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Sodium (4.5 g) was added to methanol (15 ml) to obtain a methanol solution of sodium methylate was added EM (195,4 mg) and iodine (353,6 mg) in this order at 0°and was stirred for 3 hours After confirming the completion of reaction by TLC) was added sodium thiosulfate (0.8 g), aqueous ammonia (0.5 ml) and water (80 ml) and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and the solvent drove away with getting wet substances. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (166,3 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-ethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (26,6 μl) and ethyliodide (12,2 ml) was added to a solution of AM (21,0 mg) in dimethylformamide (1.0 ml) and stirred at room temperature is within 4 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (7,0 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-diethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (26,6 μl) and ethyliodide (12,2 ml) was added to a solution of AM (21,0 mg) in dimethylformamide (1.0 ml) and stirred at room temperature for 4 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to)- receiving EM (10,3 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-allyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Allylbromide (148,3 ml) was added to a solution of AM (117,8 mg) and N,N-diisopropylethylamine (298,6 μl) in dichloromethane (5,7 the l) at 0° C and was stirred at room temperature for 2 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (to 21.9 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-diallyl-8,9-anhydrous-pseudoalteromonas A 6,9-hemiketal (EM)

Allylbromide (148,3 ml) was added to a solution of AM (117,8 mg) and N,N-diisopropylethylamine (298,6 μl) in dichloromethane (5.7 ml) at 0°and was stirred at room temperature for 2 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol: aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (64,3 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-acetyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Acetic anhydride (8,4 ml) was added the solution UM (34.8 mg) in dichloromethane (1.6 ml) at 0° C, was stirred for 10 min and then stirred at room temperature for 30 minutes After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol = 100:1,0→20:1) to obtain EM (of 33.4 mg, white powder).

Synthesis of de(3’-N-methyl)-3’-N-sulfonyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Methanesulfonanilide (9,3 ml) was added to a solution of AM (87,6 mg) in dichloromethane (4,2 ml) at 0°and was stirred for 3 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol = 100:1→20:1) to obtain EM (37,2 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-propargyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

3-Bromopropene (137,8 ml) was added to a solution of AM (106,3 mg) and N,N-diisopropylethylamine (269,3 μl) in di is loretana (5,2 ml) and stirred at room temperature for 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (41.3 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

3-Bromopropene (137,8 ml) was added to a solution of AM (106,3 mg) and N,N-diisopropylethylamine (269,3 μl) in dichloromethane (5.2 ml) and stirred at room temperature for 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01→10:1:0.05 to) obtaining EM (27.9 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-propyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (59,6 μl) and 1-jumproping (33,3 μl) were added in that order to a solution EM (23,5 m is) in acetonitrile (2.3 ml) and boiled under reflux at 80° With over 20 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (5.7 mg, white powder).

Synthesis of bis-de (3’-N-methyl)-3’,3’-N,N-dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (59,6 μl)and 1-jumproping (33,3 μl) were added in that order to a solution EM (23,5 mg) in acetonitrile (2.3 ml) and boiled under reflux at 80°C for 20 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (12.0 mg, white powder).

Synthesis of bis-de (3’-N-methyl)-3’-N-benzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Benzylchloride (297,3 ml) was added to a solution of AM (88,8 mg) and N,N-diisopropylethylamine (of 450.1 µl) in chlormethine (4.3 ml) at room temperature and was stirred for 96 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (to 49.9 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-dibenzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (135,9 μl) and benzylchloride (89,7 μl) were added in that order to a solution EM (26,8 mg) in acetonitrile (1.3 ml) and boiled under reflux at 80°C for 60 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (19.6 mg, white powder).

Synthesis of de(3’-dimethylamino)-3’-piperidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (42,5 mm) and 1.5-dibromethane (33,2 μl) were added in that order to a solution EM (16,8 mg) in acetonitrile (4,9 ml) and boiled under reflux at 80° With within 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (13.3 mg, white powder).

Synthesis of de (3’-dimethylamino)-3’-pyrrolidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (40,2 ml) and 1,4-dibromobutane (27,6 μl) were added in that order to a solution EM (15,9 mg) in acetonitrile (4.6 ml) and boiled under reflux at 80°within 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:1:0,1) obtaining EM (to 11.9 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-(2-propyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (459,2 μl) and 2-bromopropane (of 247.5 μl) were added in this order to the Astaro EM (90,6 mg) in acetonitrile (4,4 ml) and stirred at 80° With within 72 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:1:0,1) obtaining EM (to 25.3 mg, white powder). Got to 47.1 mg of a crude substance EM.

Synthesis of de(3-O-cladinose)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

p-Toluensulfonate acid monohydrate (80,3 mg) was added to a solution of AM (201,6 mg) in dimethylformamide (5.6 ml) and stirred at 50°within 8 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water, brought to pH 8.0 by the addition of saturated aqueous sodium bicarbonate solution and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (84,7 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’-N-hexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (408,5 ál and 1 bromhexin (328,7 μl) were added in that order to a solution EM (80,6 mg) in acetonitrile (3.9 ml) and stirred at 60° With within 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (33,7 mg, white powder). Got to 24.6 mg of a crude substance EM.

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-dihexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (116,0 μl) and 1-bromhexin (93,6 μl) were added in that order to a solution EM (22.9 mg) in acetonitrile (1.1 ml) and stirred at 60°within 72 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (20,1 mg, white powder).

Synthesis of de (3’-N-methyl)-3’-N-(2-foradil)8.9bn-anhydro-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (347,7 μl) and 1-bromo-2-foraten (of 148.6 ml) was added to the races the thief EM (70.0 mg) in dimethylformamide (3.3 ml) at room temperature and was stirred for 48 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (36,0 mg, white powder). Got to 25.5 mg of a crude substance EM.

Synthesis of de (3’-N-methyl)-3’-N-cyanomethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (320,9 μl) and bromoacetonitrile (128,3 ml) was added to a solution of AM (64,6 mg) in dimethylformamide (3.1 ml) at room temperature and was stirred for 4 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (53,1 mg, white powder).

Synthesis of de(12-hydroxy)-de[12-(1-hydroxypropyl)]-12-oxo-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

The leads to compounds, which lead (508,0 mg) was added to a solution of AM (508,0 mg) dichlormid is not (24,0 ml) and stirred at room temperature for 40 minutes After confirming the completion of reaction by TLC, the reaction mixture was diluted with a mixture of a saturated solution of salt - water saturated solution of sodium bicarbonate (1:1, vol/about.) and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:0,5:0,01) obtaining EM (282,7 mg, white powder).

Synthesis of de(12-hydroxy)-de[12-(1-hydroxypropyl)]-12-hydroxy-oxime-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Pyridine (0.9 ml) was slowly added at 0°With the solution UM (116,5 mg) and hydroxylamine hydrochloride (32,0 mg) in ethanol (0.9 ml) and was stirred for 3 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia 10:0,5:0,01→10:1:0.05 to) obtaining EM (114,5 mg, white powder).

Synthesis of de(3’-N-methyl)-[3’-N-(3-hydroxy-1-propyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-aminobutiramida is Tramin (338,3 μl) and 3-bromo-1-propanol (covers 175.6 ml) was added to a solution of AM (68,1 mg) in dimethylformamide (3.3 ml) at room temperature and was stirred for 48 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (27.7 mg, white powder). Got to 22.5 mg of a crude substance EM.

Synthesis of de(3’-N-methyl)-3’-N-(acetoxyethyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (106,8 μl) and 2-Bromeliaceae (67,6 ml) was added to a solution of AM (21,5 mg) in dimethylformamide (1.0 ml) at room temperature and was stirred for 48 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (6.0 mg, white powder).

Synthesis de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Sodium borohydride (21,8 ml) was added to a solution of AM (37,7 mg) in methanol (2.9 ml) at -78°and lane is massively within 30 minutes The temperature of the reaction mixture was raised to 0°and then was stirred for 30 minutes After confirming the completion of reaction by TLC, the reaction was stopped by addition of acetone (0.5 ml). The reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (35,8 mg, white powder).

Synthesis of de(3’-dimethylamino)-3’-morpholino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (45,8 μl) and bis(2-bromacil)ether (33,1 μl) were added in that order to a solution EM (18,1 mg) in acetonitrile (2.6 ml) and stirred at 80°within 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (12.0 mg, white powder).

Synthesis of de(3’-dimethylamino)-3’-[hexahydro-1-(1H)-azepine]8.9bn-anhydrous-pseudoe is ithromycin And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (49,5 mm) and 1,6-dibromohexane (43,6 μl) were added in that order to a solution EM (19.5 mg) in acetonitrile (2.8 ml) and stirred at 80°within 24 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (11.7 mg, white powder).

Synthesis of bis-de(3’-N-methyl)-3’,3’-N,N-di-(10-bromo-1-decanol)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

N,N-diisopropylethylamine (45,6 μl) and 1,10-dibromodecane (58,9 μl) were added in that order to a solution EM (18,0 mg) in acetonitrile (2.6 ml) and boiled under reflux at 80°within 36 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 20:1:0,1) obtaining EM (14,9 mg, white powder).

Synthesis of de(12-g is droxi)de[12-(1-hydroxypropyl)]-12-amino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

The oxide of molybdenum (IV) (10.0 mg) and sodium borohydride (10.5 mg) was added to a solution of AM (15,5 mg) in ethanol (2.3 ml) at 0°and was stirred for 4 hours After confirming the completion of reaction by TLC, the reaction was stopped by addition of acetone (0.5 ml) and the reaction mixture was diluted with a mixture of a saturated solution of salt - water saturated solution of sodium bicarbonate (1:1, vol/about.) and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 10:1:0,1) obtaining EM (13,4 mg, white powder).

Synthesis of de(3’-N-methyl)-de(12-hydroxy)-de-[12-(1-hydroxypropyl)]-12-oxo-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

The leads to compounds, which lead (508,0 ml) was added to a solution of AM (508,0 mg) in dichloromethane (24,0 ml) and stirred at room temperature for 40 minutes After confirming the completion of reaction by TLC, the reaction was stopped by addition of acetone (0.5 ml) and the reaction mixture was diluted with a mixture of a saturated solution of salt - water saturated solution of sodium bicarbonate (1:1, vol/about.) and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and deleted Rast is oritel with getting wet substances. The crude substance was purified column chromatography on silica gel (mixture of chloroform: methanol:aqueous ammonia = 10:0,5:0,01) obtaining EM (71,6 mg, white powder).

Synthesis of de(3’-N-methyl)-de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

Sodium borohydride (22.9 ml) was added to a solution of AM (38,8 mg) in methanol (3.0 ml) at 0°and was stirred for 1 h After confirming the completion of reaction by TLC, the reaction was stopped by addition of acetone (0.5 ml) and the reaction mixture was diluted with a mixture of a saturated solution of salt - water saturated solution of sodium bicarbonate (1:1, vol/about.) and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (of 31.4 mg, white powder).

Synthesis of de (3-O-cladinose)-de (3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal (EM)

p-Toluensulfonate acid monohydrate (53,9 mg) was added to a solution of AM (132,4 mg) in dimethylformamide (3.8 ml) and stirred at 50°C for 6 hours After confirming the completion of reaction by TLC, the reaction mixture was diluted with water, brought to pH 8.0 by addition of a saturated aqueous solution of bicarbona the sodium and was extracted with dichloromethane. The organic layer was obezvozhivani the addition of mirabilite, filtered to remove mirabilite and solvent was removed to obtain the crude material. The crude substance was purified column chromatography on silica gel (mixture of chloroform:methanol:aqueous ammonia = 15:1:0,1) obtaining EM (50.2 mg, white powder).

Brief description of figures

Figure 1(A) shows the protein P24 in the culture supernatant in M-Mϕ and GM-Mϕinfected with a strain of HIV-1JR-FLand (C) shows the protein P24 in the culture supernatant of M-Mϕ and GM-Mϕinfected with a strain of HIV-1BAL.

Figure 2(A) shows cytopathy M-Mϕinfected with a strain of HIV-1JR-FLand strain of HIV-1BALand (C) shows cytopathy GM-Mϕinfected with a strain of HIV-1JR-FLand strain of HIV-1BAL.

Figure 3(a) shows the intracellular distribution of the protein P24 in M-Mϕinfected with a strain of HIV-1JR-FLand strain of HIV-1BALand (C) shows the intracellular distribution of the protein P24 in GM-Mϕinfected with a strain of HIV-1JR-FLand strain of HIV-1BAL.

Figure 4(A) shows the detection of viral DNA in M-Mϕ and GM-Mϕinfected with a strain of HIV-1JR-FLand (C) shows the detection of viral DNA in M-Mϕ and GM-Mϕinfected with a strain of HIV-1BAL.

Figure 5 shows the changes in the expression of protein tyrosine kinase Hck in M-Mϕ and M-Mϕ and expression when HIV-1BAL.

Figure 6 shows the changes in the expression of protein/HEBβ M Mϕ and GM-Mϕ and expression in M-Mϕ or GM-Mϕinfected with a strain of HIV-1BAL.

Figure 7(a) shows the growth response of the detection of the protein P24 in the culture supernatant after infection with HIV-1 strainBALin human alveolar Mϕ (a-Mϕand (b) shows the detection of viral DNA.

On Fig shows the changes in the expression of protein tyrosine kinase Hck and protein/HEBβ alveolar Mϕ and the expression upon infection with HIV-1 strainBAL.

Figure 9 shows the expression of protein tyrosine kinase Hck in M-Mϕprocessed antimyeloma oligonucleotide to protein tyrosine kinase Hck before and after infection with HIV-1 strainBAL.

Figure 10 shows the inhibition of proliferation of HIV-1 in M-Mϕprocessed antimyeloma oligonucleotide to protein tyrosine kinase Hck.

Figure 11 shows the expression of protein/HEBβ GM-Mϕprocessed antimyeloma oligonucleotide to protein/HEBβ before and after infection with HIV-1 strainBAL.

On Fig shows the proliferation of HIV-1 in GM-Mϕprocessed antimyeloma oligonucleotide to protein/HEBβ.

On Fig shows the effect of macrolide derivatives on the proliferation of HIV-1 in M-Mϕ adding macrolide is derived, such as EATING, EM, EM, EM or HIMSELF, in comparison with the addition of DMSO.

On Fig shows the effect of macrolide derivatives on the proliferation of HIV-1 in GM-Mϕ adding a macrolide derivatives, such as EATING, EM, EM, EM or HIMSELF, in comparison with the addition of DMSO.

On Fig shows the effect of macrolide derivatives on the proliferation of HIV-1 in M-Mϕreceived from different people (three subjects) adding a macrolide derivatives, such as EATING, EM, EM, EM or HIMSELF, in comparison with the addition of DMSO.

On Fig shows the effect of the concentration of macrolides, such as EATING, EM, EM, EM HIMSELF, on the proliferation of HIV-1 in M-Mϕ compared with the addition of DMSO.

On Fig shows the effect of macrolide derivatives on cytopathy M-MϕHIV-1, adding EAT EM, EM, EM or compared with the addition of DMSO.

On Fig shows the effect of macrolide derivatives, such as EATING, EM, EM, EM or HIMSELF, on the expression of protein tyrosine kinase Hck in M-Mϕinfected with HIV-1.

On Fig(A) shows the effect of macrolide derivatives, such as EATING, EM, EM, EM or HIMSELF, to activate RAC in M-MϕHIV-1, compared with DMSO and (B) shows the effect of macrolide derivatives, such as EATING, EM, EM, EM or HIMSELF, on the activation of ERK1/2 in M-MϕHIV-1, compared with DMSO.

On pig show is but the effect of the inhibitor MRK (SB203580) and an inhibitor of ERK1/2 (PD98059) on the proliferation of the virus in M-Mϕ infected with a strain of HIV-1BLcompared with the control group.

On Fig shows the effect of different concentrations of macrolides, such as EM, EM, EM or EM, on the proliferation of HIV-1 in M-Mϕ compared with DMSO.

On Fig shows the effect of different concentrations of macrolides, such as EM, EM, EM or EM, on the proliferation of HIV-1 in M-Mϕ compared with DMSO.

On Fig shows the effect of different concentrations of the macrolide, such as EM, on the proliferation of HIV-1 in M-Mϕ compared with DMSO.

On Fig shows the effect of different concentrations of macrolides, such as EM, EM or EM, on the proliferation of HIV-1 in M-Mϕ compared with DMSO.

On Fig shows the effect of different concentrations of the macrolide, such as EM, on the proliferation of HIV-1 in M-Mϕ compared with DMSO.

A detailed description of the preferred embodiments

The present invention will be explained specifically with the help of the following examples, but it is not limited to these examples.

To show the inhibitory effect of the present invention on the growth, macrolide derivatives, EAT, EM, EM, EM ITSELF, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM or EM was dissolved in DMSO, each at a concentration of 100 mm and used at a dilution medium. The control group was M-Mϕprocessed only by the MCO, used for dissolution of macrolide derivatives.

M-Mϕ and GM-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, was added 30 μm EAT EM, EM, EM ITSELF, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM or EM, were incubated for 14 days and determined the amount of protein P24 in the culture supernatant, depending on the time.

Example 1

Effect EAT EM, EM, EM or HIMSELF on the proliferation of HIV-1 in M-Mϕ and GM-Mϕ.

Compared with the control group (DMSO) adding AM or AM protein P24 in M-Mϕ almost not detected, and the replication of the virus was significantly suppressed on day 14 of cultivation (see Fig). In M-Mϕ adding ITSELF was established almost half suppressed, and in the group with addition of EAT or EM was also observed suppression of reproduction, although not as strong compared to EM and EM (see Fig). In GM-Mϕ in the group with addition of EAT, EM, EM, EM or almost not been established whether the reproduction of the virus in all study periods, similar to the control group (DMSO) (see Fig). In the experience of using M-Mϕderived human monocytes taken from three adult volunteers received the same results as shown on Fig (see Fig).

The inhibitory effect EM and EM in the HIV-1 BALin M-Mϕ concentration dependent, and the replication of the virus was completely suppressed at a concentration of 3 μm or above (see Fig). Although the proliferation of the virus was observed at a concentration of 300 μm, not only for EM and EM, but also to EAT, EM ITSELF, and because it was observed in the control group (DMSO), possibly due to the cytotoxicity of DMSO used as a solvent, further experiments were performed at a concentration of 30 ám.

Example 2

Effect EAT EM, EM, EM HIMSELF on cytopathy M-Mϕinfected with HIV-1.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, was added 30 μm EAT EM, EM, EM or HIMSELF, incubated and examined the morphology of the cells. In the control group (DMSO) and the group with the addition of EAT, EM or MYSELF have found little education MGC, coinciding with the increase in the number of protein P24 as shown in example 1, but in the group with addition of EM and IM not observed manifestations of the cytopathic effect, such as education MGC (see Fig).

Example 3

Effect EAT EM, EM, EM HIMSELF on protein tyrosinekinase Hck in M-Mϕinfected with HIV-1.

As described previously, M-Mϕ effectively expressed protein to tyrosinekinase Hck, and regulation of expression of protein tyrosine kinase Hck by using antisense oligonucleotides can inhibit the proliferatio HIV-1 M-Mϕ . This result indicated that the expression of protein tyrosine kinase Hck is required for the proliferation of HIV-1 in M-Mϕ. Because EM and EM inhibited the proliferation of HIV-1 in M-Mϕ, it has been suggested that the expression of protein tyrosine kinase Hck was suppressed in M-Mϕtreated with these agents.

Therefore, M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, was added 30 μm EAT EM, EM, EM or HIMSELF, and incubated for 2 days after infection was assessed by the expression of protein tyrosine kinase Hck Western blot testing. In the group with making EATING, EM and MYSELF who were not very strong overwhelming effect on the proliferation of the virus was observed reduced levels of protein tyrosine kinase Hck, although not as strong compared with the control group (treated only DMSO) (see Fig). In experiments with EM and EM, which showed a strong inhibitory effect on the proliferation of the virus, expression of the protein tyrosine kinase Hck was largely suppressed (see Fig).

Example 4

Effect EAT EM, EM, EM or activation RMR and ERK1/2 in M-Mϕinfected with HIV-1.

RMR and ERK1/2 (R42 cable line/ARC) participate in various reactive mechanism of intracellular signal transduction. Phosphorylation of tyrosine RMR and ERK1/2 in M-Mϕinfected with a strain of HIV-1 , were evaluated by Western blotting. In the result, it was found that the phosphorylation of tyrosine RMR was significantly enhanced under the influence of virus infection, but the tyrosine phosphorylation ERK1/2 was weak. These results indicate that activation RAC is important for the proliferation of the virus in M-Mϕ.

Therefore, studied the effect of EATING, EM, EM, EM ITSELF, which have a overwhelming effect on the proliferation of the virus in M-Mϕon activation RMR and ERK1/2.

30 μl EAT EM, EM, EM or added to M-Mϕinfected with a strain of HIV-1BALthe phosphorylation of tyrosine RMR was determined on 2 days of cultivation Western blot testing. In the group with EATING, EM and MYSELF who were not very strong overwhelming effect on the proliferation of the virus was observed reduced phosphorylation of tyrosine RMR, although not as strong compared with the control group (DMSO). In the group with addition of EM and EM, which showed a strong inhibitory effect on the proliferation of the virus, the phosphorylation of tyrosine RMR was significantly reduced [see Fig(A)]. In all groups the total number REC (the total amount of phosphorylated and nefosfaurilirovanna RMR) was equal. Based on these results it can be assumed that the suppression of cell proliferation of the virus under the influence of EAT, EM, M, EM HIMSELF in M-Mϕ associated with lower levels RMR.

On the other hand, in the group with addition of EAT, EM and MYSELF who were not very strong overwhelming effect on the proliferation of the virus, the tyrosine phosphorylation ERK1/2 weakly detected, as in the control group (DMSO), but in the group with addition of EM and EM, which show a strong overwhelming effect on the proliferation of the virus, found increased tyrosine phosphorylation ERK1/2. In all groups the total amount of ERK1/2 (total amount of phosphorylated and nefosfaurilirovanna RMR) was equal to [see Fig ()].

Example 5

The effect of inhibitor MRK on the proliferation of the virus in M-MϕHIV-1, the suppression of the addition of EAT, EM, EM, EM ITSELF, used in the present invention.

Based on the results of example 4 it has been suggested that activation RAC is important for the proliferation of HIV-1 in M-Mϕand the overwhelming influence of EAT, EM, EM, EM HIMSELF on the proliferation of the virus is the result of the inhibitory effect of these compounds on the activation of RAC.

Therefore, the inhibitor MRK, SB203580, (4-[4-forfinal]-2-[4-methylsulfinylphenyl]-5-[4-pyridyl]-1H-imidazole) and the inhibitor of ERK1/2, PD98059, (2-[2-amino-3-methoxyphenyl]-4H-1-benzopyran-4-one), at a concentration of 10 μm, was added to M-Mϕinfected with a strain of HIV-1BAL , and studied the effects on the virus.

When 10 μm SB203580 was added to M-Mϕinfected with a strain of HIV-1BALand incubated protein P24 almost did not even 14 days, and proliferation of the virus was suppressed (see Fig). On the other hand, when added to 10 μm inhibitor of ERK1/2, PD98059, the amount of protein P24 did not differ from the control group, and was discovered proliferation of the virus (see Fig). From the results of these experiments suggest that, for the proliferation of HIV-1 strain-directed to macrophages, basically, activation of protein RAC, and it is believed that the involvement of ERK1/2 is negligible.

Example 6

Influence EM, EM, EM and EM used in the present invention, on the proliferation of HIV-1 in M-Mϕ.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, added EM, EM, EM or EM in various concentrations, were incubated for 10 days and determined the protein P24 in the culture supernatant, depending on time. The group treated with only DMSO used to dissolve the macrolide derivatives, served as a control group.

The increase in the number of protein P24 was observed in the culture supernatant in the control group when adding only DMSO depending on the duration of cultivation in the day, and was discovered Strait is ferocia HIV-1. However, compared with the control group in the group with addition of EM, EM, EM or EM, the formation of a protein P24 was reduced depending on the concentration of the agent, and was found to suppress proliferation of the virus. In particular, it was found that at a concentration of 30 μm any agents EM, EM, EM and AM almost completely inhibited the proliferation of the virus (see Fig). With agents EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM and EM got similar results as Fig. In addition, experience with the use of M-Mϕderived human monocytes taken from three adult healthy volunteers received the same results as Fig.

Example 7

Influence EM, EM, EM and EM used in the present invention, on the proliferation of HIV-1 in M-Mϕ.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, added EM, EM, EM and EM in various concentrations, were incubated for 10 days and determined the protein P24 in the culture supernatant, depending on time. The group treated with only DMSO used to dissolve the macrolide derivatives, served as a control group.

The increase in the number of protein P24 was observed in the culture supernatant in the control group when new is only DMSO depending on the duration of cultivation in the day, and was detected proliferation of HIV-1. However, compared with the control group in the group with addition of EM, EM, EM and EM, the formation of a protein P24 was reduced depending on the concentration of the agent, and was found to suppress proliferation of the virus. In particular, it was found that at a concentration of 30 μm any agents EM, EM, EM and AM almost completely inhibited the proliferation of the virus (see Fig). In addition, experience with the use of M-Mϕderived human monocytes taken from three adult healthy volunteers received the same results as Fig.

Example 8

Influence EM used in the present invention, on the proliferation of HIV-1 in M-Mϕ.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, added EM in various concentrations, were incubated for 10 days, and protein content was determined P24 in the culture supernatant, depending on time. The group treated with only DMSO used to dissolve the macrolide derivatives, served as a control group.

The increase in the number of protein P24 was observed in the culture supernatant in the control group when adding only DMSO depending on the duration of cultivation in the day, and was detected proliferation of HIV-1. However, compared to control who enoy group with the addition of EM the formation of a protein P24 was reduced depending on the concentration of the agent, and was found to suppress proliferation of the virus. In particular, it was found that at a concentration of 30 μm AM almost completely inhibited the proliferation of the virus (see Fig). In addition, experience with the use of M-Mϕderived human monocytes taken from three adult healthy volunteers received the same results as Fig.

Example 9

Influence EM, EM and EM used in the present invention, on the proliferation of HIV-1 in M-Mϕ.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, added EM, EM and EM in various concentrations, were incubated for 10 days and determined the protein P24 in the culture supernatant, depending on time. The group treated with only DMSO used to dissolve the macrolide derivatives, served as a control group.

The increase in the number of protein P24 was observed in the culture supernatant in the control group when adding only DMSO depending on the duration of cultivation in the day, and was detected proliferation of HIV-1. However, compared with the control group in the group with addition of EM, EM and EM the formation of a protein P24 was reduced depending on the concentration of the agent, and was found to suppress proliferation of the virus. In particular, it was the mouth of the lished, that at a concentration of 30 μm agents EM, EM and AM almost completely inhibited the proliferation of the virus (see Fig). In addition, experience with the use of M-Mϕderived human monocytes taken from three adult healthy volunteers received the same results as Fig.

Example 10

Influence EM used in the present invention, on the proliferation of HIV-1 in M-Mϕ.

M-Mϕ was subjected to contact exposure to HIV-1 strainBALwithin 2 h, added EM in various concentrations, were incubated for 10 days and determined the protein P24 in the culture supernatant, depending on time. The group treated with only DMSO used to dissolve the macrolide derivatives, served as a control group.

The increase in the number of protein P24 was observed in the culture supernatant in the control group when adding only DMSO depending on the duration of cultivation in the day, and was detected proliferation of HIV-1. However, compared with the control group in the group with addition of EM the formation of a protein P24 was reduced depending on the concentration of the agent, and was found to suppress proliferation of the virus. In particular, it was found that at a concentration of 30 μm AM almost completely inhibited the proliferation of the virus (see Fig). The AOC is e, in the experience of using M-Mϕderived human monocytes taken from three adult healthy volunteers received the same results as Fig.

As described above, it was found that macrolide derivatives used in the present invention, have an overwhelming impact on the proliferation of HIV-1, directed to the macrophages, M-Mϕderived from human monocytes. In particular, EM and EM, which were found to have activity effectively suppress the proliferation of HIV-1, directed to the macrophages, M-Mϕalmost completely inhibited the proliferation of the virus even at a concentration of 3 μm. It was found overwhelming impact EM and EM on the proliferation of the virus with the overwhelming manifestation of a high efficiency, equal to 95% or more, even at 14 days after incubation, only when making EM or EM in primary culture medium after contact infection.

In addition, EAT, EM and he also showed an overwhelming impact on the proliferation of the virus, depending on the concentration of the drug, and was installed the same overwhelming effect EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM, EM and EM at a concentration of 30 μm with nearly complete inhibition. Proceeding from this fact, it was found that the inhibitory effect is included in farmakologicheskie properties of macrolides, accumulating in tissue macrophages with a long validity period.

As described previously, the expression of protein tyrosine kinase Hck is important for the proliferation of HIV-1 in M-Mϕand because different macrolide derivatives according to the present invention inhibited the expression of protein tyrosine kinase in M-Mϕ, it has been suggested that this inhibitory effect is one of the mechanisms of the overwhelming action of these compounds on the proliferation of HIV-1. In addition, since various macrolide derivatives according to the present invention, showing the inhibitory effect on the proliferation of the virus, inhibited phosphorylation of tyrosine RMA, it has been suggested that the suppression of cell proliferation in HIV-1-based suppression of activation RMA data connections.

Industrial applicability

As described above, the present invention relates to the use of macrolide derivatives to suppress the infection and proliferation of human immunodeficiency virus in macrophages derived from human monocytes. The present invention is not only suitable as drugs to suppress the infection and proliferation of human immunodeficiency virus, but it is also possible their use in the clinic as an additional drug in HAART.

References

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1. Drug to suppress the infection and proliferation of human immunodeficiency virus in macrophages derived human monocytes, including derivative of erythromycin.

2. The drug according to claim 1, characterized in that the macrophages derived human monocytes represent macrophages M type M

3. The drug according to claim 1, characterized in that the inhibitory effect on the proliferation of the virus is a suppression of the expression of protein tyrosine kinase Hck and suppression of activation RMR in macrophages M type M

4. The drug according to claim 1, wherein the erythromycin derivative is any one of oxacyclohexadecan-2,10-dione,4[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexo iranoil)oxy]-14-ethyl-7,12,13-trihydroxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β -D-Xylo-hexopyranosyl]oxy]; 11-(1’-hydroxypropyl)-3-[[2,6-dideoxy-3-C-methyl-α-L-RIBO-hexopyranosyl]oxy]-5-[(3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl)oxy]-2,4,6,8,11,14-HEXAMETHYL-10,13,15-three-ocatillo [9.2.1.1.9.6]pentadecane-1-it;

6,15,16-trioxadecyl [10.2.1.11.4]hexadecane; 4,13-dioxabicyclo [8.2.1]tridec-12-EN-5-it,7-[(2,6-dideoxy-3-C-methyl-α-L-abovecaptionskip)oxy]-3-(1,2-dihydroxy-1-methylbutyl)-2,6,8,10,12-pentamethyl-9-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy]; oxazolo-tetradecane-2,10-dione; 4-[(2,6-dideoxy-3-O-methyl-α-L-RIBO-hexopyranosyl)oxy]-14-ethyl-12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy];

de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(3’-N-methyl)-3’-N-sulfonyl-6,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(3’-N-methyl)-[3’-N-(3-hydroxy-1-propyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(3’-N-methyl)-3’-N-(2-acetoxyethyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(3’-N-methyl)-3’-N-cyanomethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(3’-N-methyl)-3’-N-(2-foradil)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6,9-hemiketal or gasoli;

bis-de(3’-N-methyl)-3’-N-ethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3’-N-methyl)-3’,3’-N,N-diethyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3’-N-methyl)-3’-N-allyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3’-N-methyl)-3’,3’-N,N-diallyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3’-N-methyl)-3’-N-propargyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3’-N-methyl)-3’,3’-N,N--dipropyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3'-N-hexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt; bis-de(3'-N-methyl)-3',3'-N,N-dihexyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3'-N-benzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3',3'-N,N-dibenzyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3'-N-(2-propyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3',3'-N,N-di-(10-bromo-1-decanol)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

bis-de(3'-N-methyl)-3'-N-acetyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal Il its salts; de(3'-dimethylamino)-3'-piperidino-8,9-ang the draw pseudoalteromonas And 6.9-hemiketal or its salts; de(3'-dimethylamino)-3'-pyrrolidino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de(3'-dimethylamino)-3'-morpholino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de(3'-dimethylamino)-3'-[hexahydro-1-(1H)-azepine]8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de(12-hydroxy)-de[12-(1-hydroxypropyl)]-12-hydroxyacyl-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts; de(12-hydroxy)-de-[12-(1-hydroxypropyl)]-12-amino-8,9-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de(3’-N-methyl)-de[12-(1-hydroxypropyl)] - 8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salt;

de(3-O-cladinose)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts and de(3-O-cladinose)-de(3’-N-methyl)8.9bn-anhydrous-pseudoalteromonas And 6.9-hemiketal or its salts.

Convention priority from 07.03.2002 (JP 2002-61788) according to claims 1-3 and 4 in terms of signs: compounds listed from the beginning to “oxazolo-tetradecane-2,10-dione-4-[2,6-dideoxy-3-O-methyl-α-L-ribono-hexopyranosyl)oxy]-14-ethyl-12,13-dihydroxy-7-methoxy-3,5,7,9,11,13-HEXAMETHYL-6-[[3,4,6-trideoxy-3-(dimethylamino)-β-D-Xylo-hexopyranosyl]oxy]” inclusive, and all other compounds according to claim 4 have priority from 06.09.2002(JP 2002-261024).



 

Same patents:

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new aminobenzophenones of the formula (I):

or their pharmaceutically acceptable salts. These compounds elicit properties of inhibitors of cytokines secretion, in particular, 1β-interleukin (IL-1β) and tumor necrosis α-factor (TNF-α) and to secretion of polymorphonuclear superoxide that are useful for treatment of inflammatory diseases, for example, skin diseases, such as psoriasis, atopic dermatitis. In the formula (I) R1 is taken among the group consisting of halogen atom, hydroxy-, mercapto-group, trifluoromethyl, amino-group, (C1-C3)-alkyl, (C2-C3)-olefinic group, (C1-C3)-alkoxy-, (C1-C3)-alkylthio-, (C1-C6)-alkylamino-group, (C1-C3)-alkoxycarbonyl, cyano-group, carbamoyl, phenyl or nitro-group under condition that when R1 means a single substitute then it at ortho-position, and when R1 means more one substitute then at least one substitute of R1 is at ortho-position; R2 means one substitute at ortho-position being indicated substitute is taken among the group consisting of (C1-C3)-alkyl, (C1-C3)-alkoxy-group; R3 means hydrogen, halogen atom, hydroxy-, mercapto-group, trifluoromethyl, amino-group, (C1-C3)-alkyl, (C2-C3)-olefinic group, (C1-C3)-alkoxy-, (C1-C3)-alkylthio-, (C1-C6)-alkylamino-group, (C1-C3)-alkoxycarbonyl, phenyl, cyano-, carboxy-group or carbamoyl; R4 means hydrogen atom or (C1-C3)-alkyl; Q means a bond or -SO2-; Y means (C1-C15)-alkyl, (C3-C10)-carbocyclic group or phenyl being each of them can be substituted optionally with one or some similar or different substitutes designated by the formula R5; R5 means halogen atom, (C1-C4)-alkyl, amino-, (C1-C3)-alkoxy-group, (C1-C3)-alkoxycarbonyl or -COOH; X means oxygen or sulfur atom. Also, invention relates to a pharmaceutical composition and to a method for treatment and/or prophylaxis of inflammatory diseases.

EFFECT: valuable medicinal properties of compounds and composition.

9 cl, 2 sch, 2 tbl, 29 ex

FIELD: organic chemistry, medicine, virology.

SUBSTANCE: invention relates to technology of organic compounds, namely, to 5'-aminocarbonylphosphonates d4T that are inhibitors of the human immunodeficiency virus reproduction. Invention describes 5'-aminocarbonylphosphonates d4T of the general formula: wherein R' means hydrogen atom (H), alkyl, aryl; R'' means hydrogen atom (H), alkyl, aryl; R', R'' mean cyclic alkyl; R means alkyl. These compounds are inhibitors of the human immunodeficiency virus reproduction. Invention provides preparing new compounds eliciting valuable biological properties.

EFFECT: valuable medicinal properties of compounds.

2 dwg, 1 tbl, 5 ex

FIELD: organic chemistry, biochemistry, medicine, pharmacy.

SUBSTANCE: invention relates to new bis-tetrahydrofuranbenzodioxolyl sulfonamide compounds of the formula (I): , its salts, stereoisomers and racemates that are effective inhibitors of protease activity. Also, invention relates to pharmaceutical preparations, methods for inhibition of retrovirus proteases, in particular, to resistant retrovirus proteases, to many drugs, methods for treatment and prophylaxis of infection or disease associated with retrovirus infection in mammals and to methods for inhibition of retrovirus replication. Invention provides preparing new derivatives of bis-tetrahydrofuranbenzodioxalyl sulfonamides eliciting the valuable pharmaceutical properties.

EFFECT: valuable medicinal properties of compound and composition, improved treatment method.

16 cl, 2 dwg, 3 tbl

FIELD: organic chemistry, biochemistry, medicine.

SUBSTANCE: invention relates to phosphoramidates of nucleoside analogs comprising 2',3'-dideoxy-2',3'-didehydrothymidine 5'-phosphodimorpholidate of the formula (I) and phosphoramidates of 3'-azido-3'-deoxythymidine of the formula (II) and the formula (III) that inhibit activity in reproduction of human immunodeficiency virus (HIV). Compounds are resistant to effect of dephosphorylating enzymes and able to penetrate into cells and elicit the selective activity in inhibition of DNA biosynthesis catalyzed by HIV-reverse transcriptase.

EFFECT: valuable medicinal and biochemical properties of nucleoside analogs.

4 dwg, 1 tbl, 5 ex

and tnf-" target="_blank">

The invention relates to new aminobenzophenone General formula (I)

where R1and R3designate one or more identical or different substituents selected from the group consisting of halogen, (C1-C3)-alkyl, (C1-C3)-alkoxy; provided that, if R1denotes one Deputy, he is in the ortho-position, and if R1refers to several substituents, at least one substituent R1located in the ortho-position; and R2denotes one substituent in the ortho-position, and this Deputy is selected from the group consisting of halogen, (C1-C3)-alkoxy; and R3can additionally denote hydrogen; R4represents hydrogen; X represents oxygen; Q represents -(CO)- or a bond; Y represents (C5-C15)alkyl, (C2-C15)olefinic group; and any of these groups may be optionally substituted by one or more identical or different substituents selected from the group consisting of substituents of formula R5defined below, except that when Q represents a bond, then Y appears lcil, substituted by one or more substituents selected from the group R5; or a group of formula - (Z-O)n- Z, where Z is a (C1-C3)alkyl, n is an integer >1, and the number of atoms in a continuous linear sequence of atoms in the group Y does not exceed 15; R5denotes halogen, hydroxy, amino, (C1-C6)-alkylamino, (C1-C3)alkoxycarbonyl, -COOH, -CONHR' or-COONR'R' R' means (C1-C3)alkyl; or its pharmaceutically acceptable salt

The invention relates to a derivative of hemin or their pharmaceutically acceptable salts and inhibitors of proteolytic enzymes, which are the compounds of General formula (I)

where R1and R2- substituents, which may represent amino acids, derivatives of amino acids, peptides, consisting of 1-15 amino acid residues, derived peptides consisting of 1-15 amino acid residues, and-carboxyl group of amino acids or peptides and side groups of amino acids or peptides can be modified, and it is possible that R1=R2or R1R2=OH; carboxyl group of the porphyrin can be modified methyl or other C2-C8-ester or a physiologically acceptable salt; Y-represents Cl-CH3SOO-; Me represents Fe, with the exception of compounds where

Me=Fe3+, Y-=Cl-,

R1=-LeuLeuValPheOMe, R2=-OH; R1=-ValPheOMe, R2=-OH; R1=-LeuHisOMe,

R2=-OH; R1=-LeuHisAlaOMe, R2=-OH; R1=-LeuHisNHC10H20COOMe, R22=-LeuHisNHC10H20COOMe; R1=-Lys(Tfa)AlaAlaOMe, R2=-OH;

R1=-ValPheOMe, R2=-LeuHisOMe; R1=-LeuLeuValPheOMe, R2=-LeuHisOMe;

R1=-LeuLys(Tfa)LeuOMe, R2=-OH; R1=-LeuLys(Tfa)LeuOMe, R2=-LeuHisOMe;

R1=-Lys(Tfa)AlaAlaOMe, R2=-AlaHisLys(Cbz)LeuOMe; R1=-GlyOBzl,

R2=-GlyOBzl; R1=-HisOMe, R2=-HisOMe; R1=-LeuHisOMe, R2=-LeuHisOMe;

R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-LeuHisLeuGlyCys(Bzl)OBzl;

R1=-LeuHisOMe, R2=-OEt; R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-OEt; R1=-OBzl,

R2=-OBzl; R1=-OBzl, R2=-OH; R1=-AlaOMe, R2=-OBzl; R1=-HisOMe, R2=-OBzl;

R1=-LeuHisOMe, R2=-OBzl; R1=-LeuHisLeuGlyCys(Bzl)OBzl, R2=-OBzl;

R1=-LeuHisAlaLys(Cbz)GlyCys(Bzl)OBzl, R2=-OBzl; R1=-LeuHisLys(Cbz)OMe,

R2=-OH; R1=-LeuHis(Bzl)Lys(Cbz)OMe, R2=-OH; R1=-LeuHisOMe, R2=-OMe;

R1=-LeuHis(Bzl)Lys(Cbz)OMe, R2=-OMe; R1=-AlaLeuAlaPheAlaCys(Bzl)OMe,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-AlaLeuAlaPheAlaCys(Bzl)OBzl,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-LeuHisAlaLys(Cbz)Cys(Bzl)OBzl,

R2=-LeuHis(Bzl)Lys(Cbz)OMe; R1=-LeuHisOMe, R2=-OMe;

R1=-GlyProArgGlyGlyOMe, R2=-OH;

R1=-ArgProProGlyPheSer(Bzl)PheArgGlyGlyOMe, R2=-OH,

two ways to get hemin derivatives of General formula I, hemin derivatives of the formula I, formerly known above, as inhibitors of proteolytic enzymes: the HIV protease, pepsin, trypsin, chymotrypsin

The invention relates to pharmaceutical industry and relates to the creation of a means for inhibiting reproduction enveloped viruses

The invention relates to the field of medicine and pharmaceutics and relates to a composition for the prevention and treatment of HIV-1 infection, including nucleoside reverse transcriptase FROM HIV-1, representing heterocyclics(ACS/thio)anilide, a second inhibitor of HIV-1, which does not choose the same HIV-1 mutant strain, or strains, select the first compound is an inhibitor of HIV-1

The invention relates to bicyclerelated analogues of General formula (1) and the oligonucleotides on the basis thereof, containing one or more structural units of the General formula (1A), where R1represents a hydrogen atom, a protective group, the group of phosphoric acid, etc., R2is sidegroup or amino group which may be substituted, represents a purine-9-ilen, or 2-oxo-1,2-dihydropyrimidin-1-ilen group which may be substituted by one or more substituents

FIELD: medicine, pulmonology.

SUBSTANCE: one should apply lycopid to study initial level of mature T-cells (CD3+) in % and solve following discriminant equation: D = 0.840 · (CD3+), and at D value being above 29.24 one should predict positive curative effect as a result of lycopid application, and at D value being below 29.24 on should predict no positive curative effect.

EFFECT: higher efficiency of lycopid-including therapy.

2 ex

The invention relates to enriched troxerutin containing at least 92 wt.% 7,3',4'-trihydroxyethylrutoside, from 2 to 4 wt.% 5,7,3',4'-tetrahydrochloride and from 1 to 3 wt.% 7,4'-dihydroxytoluene and method thereof

The invention relates to the pharmaceutical field, in particular to solid dosage forms of the drug, with bacteriostatic action, and can be used in a variety of infections, especially respiratory, urogenital, skin and soft tissues

The invention relates to medicine, infectious diseases and can be used for the treatment of brucellosis
The invention relates to medicine, in particular to the venereal diseases, urology and gynecology

The invention relates to medicine, infectious diseases and can be used for the treatment of brucellosis

The invention relates to medicine, namely to experimental pharmacology and cardiology

The invention relates to medicine, specifically to antibacterial drug roxithromycin
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