Salts of 5'-h-phosphonate 3'- azido-3'-deoxythymidine, which are specific inhibitors of the production of human immunodeficiency virus hiv-1 and hiv-2

 

(57) Abstract:

The invention relates to Virology and relates to new biologically active compounds, namely salts 5 N-phosphonate 3'-azido-3'- deoxythymidine General formula given in the description. The new compounds possess the ability to specifically inhibit the production of human immunodeficiency virus, suggesting the possibility of their use in medicine for the treatment of AIDS. Obtaining them carried out by neutralization of 5'-H-phosphonate 3'-azido-3' deoxythymidine hydrate oxides of the respective metals in the water to a pH of 7 to 7.2. The resulting soluble salts evaporated to a small volume, which begins spontaneous crystallization and leave at 4oWith 5 to 6 hours before the end of crystallization, the amount of precipitation is filtered off, washed with small volumes of kladenets to 4oWith water and dried. The yield of the target substances is 95 - 98%. The new compounds inhibit the replication of HIV-1 and HIV-2 in cultures of human lymphocytes H9, MT-4 and PBL are little or no hygroscopic, which facilitates the creation of dosage forms based on them. 3 table.

The invention relates to Virology and relates to new biologically active compounds, and Kengo metal.

The new compounds possess the ability to specifically inhibit the production of human immunodeficiency virus, suggesting the possibility of their use in medicine for the treatment of AIDS.

Currently, there are various compounds that inhibit the production of human immunodeficiency virus. The most popular of them received the 3'azido-3'-deoxythymidine (AZT) [1].

Also known 5'-phosphonates 3'-azido-2',3'-dideoxynucleosides [2], including 5'-H-phosphonate 3'-azido-3'-deoxythymidine, which possess inhibitory activity against HIV-1 and HIV-2, but less toxic than AZT. However, the creation of dosage forms on the basis of 5'- phosphonates 3'-azido-2', 3'-dideoxynucleosides extremely difficult due to their high hygroscopicity.

The inventive salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine activity and toxicity are not inferior to the acid form 5'-H-phosphonate 3'-azido-C'-deoxythymidine, but at the same time they are characterized by a significantly lower hygroscopicity. In addition, new salt expand the range of compounds with inhibitory activity against HIV-1 and HIV-2.

New compounds are white crystalline substances, good RA is e conduct neutralization 5'-H-phosphonate 3'-azido-3'-deoxythymidine hydrate oxides of the respective metals in water to pH 7-7,2. The resulting soluble salts evaporated to a small volume, which begins spontaneous crystallization and leave at 4oC for 5-6 h before the end of crystallization, the amount of precipitation is filtered off, washed with small volumes cooled to 4oC water and dried. The yield of the target substances is 95-98%.

The purity and structure of the new compounds were confirmed by HPLC and UV - and NMR-spectroscopy.

Here are some examples of obtaining specific salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine.

Example 1. Synthesis of sodium salt of 5'-H-phosphonate 3'-azido-3'-deoxythymidine

To a solution of 166 g (0.5 moles) of 5'-H-phosphonate 3'-azido-3'-deoxythymidine in 200 ml of distilled water, cooling water with ice and stirring for 20 min, poured 100 ml (0.5 mol) of 5N. solution of hydrate of sodium oxide (qualification CHP) until the pH of the medium was 7.2. The resulting black solution was evaporated on a rotary evaporator with a bath temperature not exceeding 45oC 150-170 ml total volume and left for 6 h at 4oC. the Precipitated crystalline precipitate was separated by a glass filter, washed with 40 ml chilled to 4oC distilled water and dried in a vacuum desiccator. The output 164 g (95.4 percent) 120-7 NH2in a linear gradient KH2PO4(0.05 to 1 M) of 7.2 min, flow rate 1 ml/min

UV-spectrum (in water)max266 nm ( 9600), 1H NMR spectrum (D2O ), M. D.: 1,93 D. (3H, CH3I , JCH3-H60.5 Hz); 2.55 (2H, H2'); 4,60-3,95 m (4H, H3' + H4' + 2H5'); 6,15 t (1H, H1', JH1'H2'6 Hz); 7,66 d (1H,H6, JH6,CH31 Hz).31P-NMR-spectrum (D2O ), M. D.: 6.5 (1H, HPI , JH,P626 Hz; JP-H5'6.3 Hz, JP-H4'1,6 Hz).

Example 2. Synthesis of potassium salt of 5'-H-phosphonate 3'-azido-3'-deoxythymidine.

To a solution of 166 g (0.5 moles) of 5'-H-phosphonate 3'-azido-3'-deoxythymidine in 200 ml of distilled water, cooling water with ice and stirring for 20 min, poured 100 ml (0.5 mol) of 5N. solution of potassium hydroxide (qualification CHP) until the pH of the medium was 7.2. The resulting black solution was evaporated on a rotary evaporator with a bath temperature not exceeding 45oC 150-170 ml total volume and left for 6 h at 4oC. the Precipitated crystalline precipitate was separated by a glass filter, washed with 40 ml chilled to 4oC distilled water and dried in a vacuum desiccator. Exit 172 g (97.1 per cent), so pl. > 300oC (charring). The substance is not hygroscopic.

Retention time on the column Nucleosil 120-7 NH2in the linear gradient is ECTR (D2O ), M. D.: 1,93 D. (3H, CH3I , JCH3-H60.5 Hz); 2.55 (2H,H2'); 4,60-3,95 m (4H, H3'+H4'+2H5'); 6,15 t (1H, H1', JH1'H2'6 Hz); 7,66 d (1H,H6, JH6,CH31 Hz).31P-NMR-spectrum (D2O ), M. D.: 6.5 (1H,HPI , JH,P626 Hz; JP-5H'6.3 Hz, JP-H4'1,6 Hz).

Example 3. The synthesis of the calcium salt of 5'-H-phosphonate 3'-azido-3'-deoxythymidine.

To a solution of 166 g (0.5 moles) of 5'-H-phosphonate 3'-azido-3'-deoxythymidine in 200 ml of distilled water, cooling water with ice and stirring for 20 min, poured in 1000 ml of 0.25 mol) of 0.5 N. the pre-filtered solution of hydrate of calcium oxide (qualification CHP) until the pH of the medium to 7.0. The resulting black solution was evaporated on a rotary evaporator with a bath temperature not exceeding 45oC to about 200 ml total volume and left for 6 h at 4oC. the Precipitated crystalline precipitate was separated by a glass filter, washed with 40 ml chilled to 4oC distilled water and dried in a vacuum desiccator. The output 174 g (97.1 per cent), so pl. > 300oC (charring). The substance is not hygroscopic.

Retention time on the column Nucleosil 120-7 NH2in a linear gradient KH2PO4(0.05 to 1 M) of 7.2 min, flow rate 1 ml/min

UV-spectrum (in water)max266 nm ( 9600) 6 Hz); 7,66 d (1H,H6, JH6,CH31 Hz).31P-NMR-spectrum (D2O ), M. D.: 6.5 (1H,HPI , JH,P626 Hz; JP-5H'6.3 Hz, JP-H4'1,6 Hz). Example 4. Synthesis of barium salt of 5'-H-phosphonate 3'-azido-3'-deoxythymidine.

To a solution of 166 g (0.5 moles) of 5'-H-phosphonate 3'-azido-3'-deoxythymidine in 200 ml of distilled water, cooling water with ice and stirring for 20 min, poured in 1000 ml of 0.25 mol) of 0.5 N. the pre-filtered solution of hydrate of oxide of barium (qualification CHP) until the pH of the medium to 7.0. The resulting black solution was evaporated on a rotary evaporator with a bath temperature not exceeding 45oC to about 3300 ml total volume and left for 6 h at 48oC. the Precipitated crystalline precipitate was separated by a glass filter, washed with 40 ml chilled to 4oC distilled water and dried in a vacuum desiccator. The output 226 g (98.5 per cent), so pl. > 300oC (charring). The substance is not hygroscopic.

Retention time on the column Nucleosil 120-7 NH2in a linear gradient KH2PO4(0.05 to 1 M) of 7.2 min, flow rate 1 ml/min

UV-spectrum (in water)max266 nm ( 9600), 1H NMR-spectrum (D2O ), M. D.: 1,93 D. (3H, CH3I , JCH3-H60.5 Hz); 2.55 (2H,H2'); 4,60-3,95 m (4H, H3'+H4'+2H5'); 6,15 t (1H, H1', JH1'H2' 6.3 Hz, JP-H4'1,6 Hz).

NMR spectra were recorded on a spectrometer Bruker 250 MHz. UV-spectra were determined on a spectrophotometer Specord UV-vis M-40. HPLC was performed using column Nucleosil 120-7 NH2.

Were studied the biological properties of new compounds, namely their ability to specifically inhibit the production of human immunodeficiency virus and toxicity. The studies were performed on cultures of human lymphocytes - human lymphoblastoid cell lines of human H9/B, MT-4 and PBL. Production of virus cells was monitored in the reactions indirect immunofluorescence assay and enzyme immunoassay. The activity of compounds characterized by using the value of ED is the effective concentration that reduces the production of HIV-1 and HIV-2 by 50% toxicity using the value of TD is the concentration that reduces cell growth by 50%. Index selectivity was calculated as the ratio of AP to ED. Were used for comparison of the acid form 5'-H-phosphonate 3'-azido-3'-deoxythymidine. Experimental data are given in table. 1-3.

Thus the new compounds are the salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine-inhibit the replication of HIV-1 and HIV-2 in cultures of human lymphocytes H9, MT-4 and PBL, and around that is was mentioned inventive salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine is also within the toxicity of the acid form 5'-H-phosphonate 3'-azido-3'-deoxythymidine. At the same time, new connections are little or no hygroscopic, which facilitates the creation of dosage forms based on them.

Salts of 5'-H-phosphonate 3'-azido-3'-deoxythymidine General formula:

< / BR>
where R is the atom of alkali or alkaline earth metal, which are specific inhibitors of the production of human immunodeficiency virus HIV-1 and HIV-2.

 

Same patents:

The invention relates to new biologically active compounds-nitroxyl derivatives of azidothymidine General formula

< / BR>
where R1radical containing nitroxyl group >NO, and R2=R1or H, which possess antiviral activity against RNA-containing viruses (human immunodeficiency virus and vesicular stomatitis virus) and DNA-containing virus (cytomegalovirus)

The invention relates to the field of organic chemistry, and in particular to methods of obtaining derivatives of nucleosides, in particular to obtain 5'-0-benzoyl-2,3'-anhidrosis-thymidine
The invention relates to the chemistry of nucleosides, in particular, to obtain 3'-azido-2', 3'-dideoxythymidine (azidothymidine, AZT), used in medicine as an antiviral drug for the treatment of acquired immunodeficiency syndrome (AIDS)

The invention relates to organic chemistry, and in particular to methods for the preparation of 3'-azido-3'-deoxythymidine, having the name 1-(3'-azido-2,3'-deoxy-D-Erythro-pentofuranose)-thiamine, azidothymidine (AZT)

The invention relates to medicine, namely to kardiologii, and can be used in the treatment of chronic ischemic heart disease
The invention relates to chemical-pharmaceutical industry, namely the method for producing a non-narcotic analgesic-analgin, analgesic, anti-inflammatory and antipyretic action, hard gelatin capsules
The invention relates to a composition medicines outdoor applications and is used in medicine for the treatment of infectious and chronic diseases of the upper respiratory tract, sore throat, bronchitis, sinusitis, tonsillitis, and as a vasodilator, keratoplastics and antipruritic tools

The invention relates to medicine, specifically to therapy and for treatment of viral hepatitis B drug lamivudine, (-)-CIS-1-(2-(2-hydroxymethyl)-1,3-oxathiolan-5-yl-cytosine

The invention relates to new derivatives of 4-aseankorea General formula (1),

< / BR>
in which R is hydrogen or C1-3alkyl group; X is chlorine, bromine or iodine and- single or double bond

The invention relates to new biologically active compounds-nitroxyl derivatives of azidothymidine General formula

< / BR>
where R1radical containing nitroxyl group >NO, and R2=R1or H, which possess antiviral activity against RNA-containing viruses (human immunodeficiency virus and vesicular stomatitis virus) and DNA-containing virus (cytomegalovirus)
The invention relates to medicine, namely to methods of treatment of male infertility

FIELD: medicine.

SUBSTANCE: method involves carrying out hernia removal in intralaminar way. Posterior longitudinal ligament defect is covered with Tacho-Comb plate after having done disk cavity curettage. Subcutaneous fat fragment on feeding pedicle is brought to dorsal surface of radix and dural sac.

EFFECT: enhanced effectiveness of treatment; reduced risk of traumatic complications.

1 dwg

FIELD: medicine, obstetrics, gynecology.

SUBSTANCE: at the background of therapy conducted one should introduce derinate immunomodulator into the body of pregnant woman additionally nasally per 1-2 drops of 0.25%-solution into each nasal canal 5-8 times daily for 3-5 d and - parenterally per 5.0 ml of 1.5%-solution once daily for 3-8 d along with preparation that improves microcirculation and along with antioxidant at a certain sequence, moreover, derinate should be introduced 30-40 min after application of microcirculation-improving preparation, and antioxidant - 20-30 min after derinate's introduction. The present innovation favors decreased edemas, decreased body weight, stabilization of Macluer-Aldrich test that in its turn enables to avoid perinatal losses, decrease the risk for the development of fetoplacental insufficiency and intrauterine fetal infection.

EFFECT: higher efficiency of therapy.

1 ex, 2 tbl

FIELD: medicine, otolaryngology.

SUBSTANCE: the present innovation deals with introducing neomycin sulfate antibiotic in granules prepared by the following technique. Tablet of neomycin sulfate 1.0g should be put into a vial with 100 ml distilled water till tablet's decomposition. Then vial's content should be shaken and kept till suspension sedimentation. In a day one should take 1 ml of supernatant liquid to be put into another vial and diluted with distilled water at 1:100 ratio. This procedure should be repeated 4 times more, moreover, during the last procedure one should apply alcohol for dilution. Then one should transfer the drop of alcoholic solution into a vial with granules out of milk sugar to then shaken and kept open for 1 d till granules" drying up. The suggested preparation should be applied per 1 granule under the tongue, moreover, multiplicity and duration of the above-suggested intake should be matched individually by patient's sensitivity and obtaining the clinic effect. The method enables to improve the value of tonic threshold audiometry by about 30-50 dB, decrease perception threshold of vocal range frequencies and widen the range towards high frequencies.

EFFECT: higher efficiency of therapy.

1 dwg

FIELD: medicine, oncology.

SUBSTANCE: invention relates to a method for treatment of chronic lympholeukosis. Method involves intravenous drop and jet administration of antitumor chemopreparations and carrying out the autochemotherapy. At the 1-st and 8-th day of treatment cyclophosphan in the dose 750 mg/m2, vincristine in the dose 1.4 mg/m2 and doxorubicin in the dose 30 mg/m2 incubated with 200 ml of autoblood are administrated to patients. From the 1-st to 14-th day of treatment prednisolone is used every day in the therapeutic dose. The treatment course is repeated in 30-35 days depending on blood indices and patient state. The total treatment of courses is 4-5. Method provides reducing cardiotoxicity of doxorubicin and cumulative toxicity of chemopreparations that allows carrying out administration of antitumor chemopreparations in the full volume to patients of elderly age groups.

EFFECT: improved method for treatment.

1 ex

FIELD: medicine, oncohematology.

SUBSTANCE: the present innovation deals with treating elderly patients with chronic lympholeukosis accompanied with cardiovascular failure. The method deals with applying chemopreparations and cytoprotector. Moreover, 1 wk before the onset of chemotherapeutic therapy one should prescribe preductal at the dosage of 105 mg daily. At this background one should sample blood out of elbow vein at the volume of 200 ml into a vial with glugicir to centrifuge it, isolate plasma, divide into two portions, add into the 1st vial - cyclophosphan 600-800 mg/sq. m, vincristin 1.4 mg/sq. m, into the 2nd vial - adriamycin 50 mg/sq. m to be incubated for 30 min at 37 C and intravenously injected by drops for patients. Simultaneously, the intake of prednisolone should be prescribed at the dosage of 60 mg/sq. m since the 1st d and during the next 5 d and preductal at the dosage of 105 mg daily during a week, and then 2 wk more at the dosage of 60 mg daily. All the procedures should be repeated in above-mentioned sequence 4-6 times. The method enables to decrease toxic manifestations of chemotherapy while applying adequate dosages of cytostatics, anthracycline antibiotics, among them, at no great manifestations of their toxicity due to preductal's cardioprotective action.

EFFECT: higher efficiency of therapy.

1 ex, 5 tbl

FIELD: medicine, immunology, nucleic acids.

SUBSTANCE: invention relates to a method for stimulation of the immune response using nucleic acids-containing immunostimulating compositions, oligonucleotide-containing composition and to a method for treatment or prophylaxis of allergy or asthma. For stimulation of the immune response thymidine-enriched nucleic acid comprising poly-T sequences and/or comprising above 60% of thymidine-containing nucleotide residues is administrated. Invention provides the development of new method for stimulation of the immune response due to administration of the proposed immunostimulating nucleic acid.

EFFECT: valuable medicinal properties of nucleic acid.

27 cl, 12 dwg, 12 ex

FIELD: veterinary science.

SUBSTANCE: the suggested method should be performed under conditions of experimental modeling dystrophic process due to intraosseous introduction of glucosamine hydrochloride at the dosage of 15-25 mg/kg 1-2 times weekly during a month. The method provides high local concentrations of medicinal preparation, at its steady entering the circulation by leaving aside hepatic barrier and, as a result, optimization of chondroprotector action of glucosamine hydrochloride and better treatment of articular alterations induced in the course of an experiment in animals.

EFFECT: higher efficiency of correction.

8 dwg, 1 ex

Up!