The phosphonolipids carboxylic acids and their salts and anti-virus tool based on them

 

(57) Abstract:

The invention relates to new phosphonolipids carboxylic acids of formula I and their salts, where R1denotes a linear alkyl WITH9-C13, R2denotes a linear alkyl WITH8-C12, R3denotes hydrogen, a linear alkyl WITH1-C6or benzyl, n = 0-2, 0-3 m, which have antiviral activity and can find application in medicine. Also described antiviral agent comprising as active substance a compound of the formula I. 2 C. and 5 C.p. f-crystals, 9 PL.

The invention relates to a new lipid derived phosphonocarboxylate acids and their esters of the General formula

< / BR>
in which R1denotes a linear or branched, saturated or unsaturated alkyl chain with from 9 to 13 carbon atoms

R2denotes a linear or branched, saturated or unsaturated alkyl chain with 8 to 12 carbon atoms;

R3denotes H, a linear or branched alkyl chain with 1-6 carbon atoms, in particular methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, neopentyl, Texel or phenyl, choline, ethanolamine, carnitine, C5-C7-recloak the R5and R6denote alkyl and n = 1, 2, or 3;

n is 0,1 or 2;

m is 0, 1-3, respectively,

their tautomers, their physiologically acceptable salts with inorganic and organic bases, and also to the way they are received and containing these compounds medicines.

As compounds of General formula I contain asymmetric carbon atoms, all optically active forms and racemic mixtures of these compounds are the subject of the present invention.

Under the compounds of the formula I below includes salts, tautomers, esters, optically active forms and racemic mixtures.

Therapy of malignant neoplasms (carcinoma, sarcomas, hematological neoplasia), inflammatory diseases or autoimmune diseases, and diseases caused by viruses or retroviruses, such as, for example, AIDS, ARC (AIDS related complex), infection cytomegaly and herpes or hepatitis, along with the lack of effectiveness of therapeutic active substances, often associated with their highly undesirable side effects. This effect is due to the low selectivity in vivo, respectively limited terap the active substances, show them in vitro, often rte can be transferred to conditions in vivo.

So many years of trying by modifying the chemical structure of pharmacologically active substances to create new materials with improved therapeutic spectrum of action. In addition, often get new dosage forms of pharmaceutical drugs with the intention to purposefully to transport active substances into the place in which they should exercise their therapeutic effect. In particular, it is necessary to prevent unwanted interaction with healthy cells. One possibility of improving therapeutic range is that by a slight modification of pharmacologically active substances, for example by obtaining salt accession acid or base, or by obtaining a pharmacologically acceptable esters [for example, esters of fatty acids; J. Pharm.Sci. 79, 531 (1990)] the physical properties of the active substance change so that was improved solubility or tolerability of the active substance. These slightly chemically modified compounds are frequently referred to as "prodrugs", as they are in contact in therapeutically active agents. Such "prodrugs" of the compounds of General formula I also includes the present invention.

To improve catabolic stability carry out chemical binding of nucleosides, for example, ara-C and Agha A, with phospholipids. The corresponding derivatives are less toxic and more stable in vivo in comparison with non-modified nucleosides. However, it did not exert any influence on the absorption and permeability of cells [J. Med. Chem. , 32, 367 (1989), Cancer Res., 37, 1640 (1977) and 41, 2707 (1981)] . Other phospholipid derivatives of nucleosides is described, for example, in the following sources:

In J. Biol. Chem., 265, 6112 (1990) described the preparation and use of liposuction as antiviral drugs. However, in this case, were investigated and synthesized only associated with the known nucleosides such as AZT and ddC, dimyristoylphosphatidylcholine and dipalmitoylphosphatidyl remains of their structure esters of fatty acids.

In J. Med. Chem., 33, 1380 (1990) described nucleoside conjugates of tiefenriede with cytidinediphosphocholine, which have antitumor activity and can find application in Oncology.

In Chem. Pharm. Bull., 36, 209 (1988) described 5'-(3-SN-phosphatidyl)-Idov and phosphocholine in the presence of phospholipase D transferase activity.

Also described, in particular, in Tetrahedron Lett., 28, 199 (1987) Chem. Pharm. Bull., 36, 5020 (1988) enzymatic synthesis of liposuction.

In W094/13324 described orally applied active substance with 1-O-alkyl-, 1-O-acyl, 1-S-acyl - and 1-S-alkyl-sn-glycero-3-phosphate as a carrier lipid.

In the application EP 418814, as well as in J. Med. Chem., 34, 1912 (1991) described isoprenylcysteine as inhibitors of squalene synthetase.

In Biochem. Biophys. Res. Coinmun, 171, 458 (1990) described the lipid conjugate antiretroviral foscarnet with palmitostearate, and in J. Med. Chem. , 20, 660 (1977) have shown anti-HIV activity (hexyloxy)-hydroxyphenylarsonic acid.

In principle, significant assistance was provided by the finding efficient ways to transport therapeutic concentrations of drugs to the appropriate authorities of the target or the target cell, in the case of AIDS, for example, in cells of the immune system and the lymphatic system, which is considered the main reservoir of viral replication.

Postemergence acid (PFA=phosphonoformic acid) and phosphonooxy acid (PAA= phosphonoacetic acid) show high antiviral activity against herpes simplex virus (HSV) 1 and 2, influenza, hepatitis b virus (HBV), varicella zoster virus (VZV), the virus e their derivatives are in some cases an effective alternative, accordingly, the addition of nucleosides, as they inhibit a wide range of DNA and RNA polymerases, as well as RT retroviruses with sufficient selectivity.

Themselves postemergence and phosphonooxy acid due to their similarity with the pyrophosphate show toxicity due to their accumulation in the bones.

Compounds according to the present invention have valuable pharmacological properties. In particular, they are suitable for therapy and prophylaxis of infections caused by DNA viruses, such as herpes simplex virus, the virus cytomegaly, papovavirus, varicella zoster virus, hepatitis virus, or Epstein-Barr, influenza virus or RNA virus, such as viruses Togga, or, in particular, retroviruses, such as oncoviruses HTLV-1 and II (virus human T-cell leukemia), and lentiviruses Visna virus human immunodeficiency HIV-1 and 2.

Particularly suitable compounds of the formula I for the treatment of clinical manifestations of retroviral HIV infection in humans, such as prolonged generalized lymphadenopathy (PGL), advanced stage AIDS related complex (ARC) and the full clinical picture of AIDS-associated viral infectino action foscarnet (trinacria salt phosphonopropionic acid/ phosphonomycin acid) anti-HIV patients with CMV-retinitis.

Antiviral effects in a mouse cytomegaly virus (CMV) is described in Antiviral Res., 26, 1 (1995).

Further, in JAMA 273, 1457 (1995) described the treatment of CMV-retinitis using phosphonopropionic acid.

The conjugates of postemergence and phosphonooxy acid with 2',3'-dideoxy-3'-thiacytidine inhibiting HIV-1 replication, as described in J. Med.Chem., 37, 2216 (1994), and in J. Pharm.Sci., 83, 1269 (1994) described a complex aryloxyalkyl esters foscarnet.

Of particular interest are the application of the U.S. 5194654, respectively PCT WO 94/13682. They describe lipid derivatives phosphonocarboxylate acids and their application in the liposomes with the formation of particularly stable liposomal complex. Along with overly broad claims as the object of the application is described 1-O-alkyl-sn-glycero-3-phosphonocrotonate acids, which are particularly well integrated into the double lipid layer liposom. The stated alkyl residues can contain from 2 to 24 carbon atoms, but they are not substituted.

In the example described and confirmed by the data about antivirus action only connection 1-O-octadecyl-sn-glycero-3 - phosphonoformate (batil-phosphonoformate). In the research, and when receiving the connection had what about the substances in the solution/suspension, but not in the liposomes.

Proposed according to the present invention compounds of General formula I in the same conditions for sustainable and have both in vitro and in vivo (in a mouse model cytomegaly virus) are obvious advantages. In particular, esters of carboxylic acids in oral introduction more stable and have better bioavailability compared with the corresponding free carboxylic acids.

In an unexpected way was established a very close relationship between the effect and structure in terms of chain length used saturated alkyl residues. Only two alkyl residues with a chain length in the range of 10-13 carbon atoms gave the best effect.

Compounds that are the subject of this application, are so interesting improvement WO 94/13682 and US 5194654, that was not to be expected, and though they are covered by these applications, however, are not the object of the application, not even explicitly named, respectively, neither mentioned by name, and their application also follows clearly from the above publications.

The compounds of formula I are new. Along with higher resistance (in substance in solution) of the claimed compounds is th way it was installed, what pharmaceutically active substances of the formula I possess in comparison with the pharmacologically active unbound, respectively non-modified substances, a wider therapeutic spectrum. In addition, they improve their time in the body, bioavailability and is often known as a critical factor permeability through the membrane (e.g., blood-brain barrier, the cell membrane, and so on). The compounds of formula I are thus as a carrier system for pharmacologically active substances. The conjugates of formula I can be identified from the point of view of their function as intracellular escrow system drugs, target designation system of medicine and drug delivery. They cause pharmacologically active substance after oral administration released inside the cell, and this release is preferably not in all cells, organs or tissues of the body, but only sighting and such cells that contain a specific enzyme. However, it is particularly striking that the cleavage takes place not during the transport of the substrate in body fluids such as blood, serum or lymph fluid, or in pechenochnogo acid kidney or cleavage of the conjugate in the liver, thanks a significantly higher percentage of the active substance is transported on or in appropriate target cells. Such cells, as already mentioned above, are, in particular, physiologically or pathophysiologically activated cells, which can be used as a target for delivery of pharmacologically active substances, such as blood leukocytes, lymphocytes, macrophages and other cell populations immunology of the lymphatic system. This, in particular, is activated cells (e.g. macrophages, granulocytes, lymphocytes, leukocytes, platelets, monocytes, and so on ), which play a pathophysiological or symptomatic role in specific pathological process. In addition, we have in mind also the cells infected with viruses, bacteria, fungi, or other microorganisms.

Unexpectedly, it was also found that therapeutic range of pharmacologically active phosphonocarboxylates acid and its esters is significantly improved, if the substance is associated with a very special lepidobalanus molecule-carrier. Thus obtained conjugate serves as a new active substance to obtain the dosage forms of the active phosphonocarboxylates acid in vivo, since the resulting system of escrow, shipping and transport of the drugs causes localization of pharmacologically active substances in the target cells and thus improves the efficacy and tolerability of pharmacologically active substances. This means that, on the one hand, prescribed dosage pharmacologically active phosphonocarboxylates acid can be reduced, and on the other hand, when the same effective amount is achieved strengthening of pharmacological action.

Pharmacologically active phosphonocarboxylate acid released from the conjugate by enzymatic hydrolysis of the conjugate.

The conjugates of formula I have obvious advantages in comparison with unconjugated pharmaceutically active phosphonocarboxylates acid, respectively, with its esters. Specific covalently associated with the pharmaceutically active substance carrier improves the bioavailability of poorly rezorbiruetsa pharmaceutically active substances, tolerance to potentially toxic active molecules, time quickly removed or metabolisable medicines and penetration through the membrane compounds with poor membrane permeability of the in vivo occurs as a rule, not in the serum, but only inside cells. In addition, the component carrier improves its lecithinase structure, which is essential for the discussed effect, penetration through the membrane pharmaceutically active substances and contributes to the effect of the Deposit. In addition, gastrointestinal tolerability lipid conjugates many times better in comparison with pure pharmaceutically active phosphonocarboxylates acid. And if resorbtive lipid conjugate is better penetrates through the membrane structure and thereby overcomes better osteoclast barriers. The same applies to the penetration of, for example, through the blood-brain barrier.

Further, due to better binding of the conjugate to plasma and tissue proteins improves the distribution in vivo. In the normal biotransformation conjugate initially oxidized from tiefer (n=0) in the sulfoxide (n= 1), which, however, due to the equal action sulfoxide in comparison with tieferen is not a disadvantage. Slow release of pharmaceutically active phosphonocarboxylates acid of the conjugate provides a low, but constant for a long period of time the active substance, and thereby str substance in the form of monophosphate due to its high hydrophilic properties no longer leaves the cell.

The half-lives of pharmaceutically active substances in the organism as a whole and in cell bodies due to conjugation significantly increase because of longer residence time of the conjugate in the body. Because there is no splitting activity in serum and in various organs, it is almost not observed or observed only a very slight toxicity to bone marrow and organs. In particular, the advantage is that the conjugates of formula I specifically accumulate in various target organs, tissues or cells.

The compounds of formula I can be used as active substances to obtain drugs used to treat those diseases which are required or useful high levels of pharmaceutically active substances in cells, organs or tissues. Essential to this system, referred to as the escrow system, delivery and targeting of drugs, is that interesting in the sense of planned therapy cells have an enzyme cleavage, so that at the first stage, the active substance is bound and then transposed through the cell membrane into the cell, and the splitting of the military transport across the cell membrane, or later partly inside the cell. Intracellular cleavage occurs, in particular, in cases where the enzyme cleavage also localized within the cell.

Suitable target cells are, in particular, the immunological cells of the lymphatic system (e.g., peripheral blood leukocytes, monocytes, macrophages, lymphocytes) or infected cells.

Unexpectedly, it has also been found that compounds of General formula I inhibit the reproduction of DNA, respectively RNA viruses at the stage of virus-specific DNA, respectively RNA transcription. These substances can have through inhibition of the enzyme reverse transcriptase impact on the reproduction of retroviruses (see Proc. Natl. Acad.Sci USA 83, 1911, 1986 or Nature 325, 773, 1987). Special therapeutic interest inhibitory effect on HIV-the virus that causes the disease is human immunodeficiency (AIDS). For the treatment of AIDS currently admitted 3'-azido-3'- deoxythymidine (DE-A-3608606). However, toxic side effects of 3'-azido-3'-deoxythymidine on the bone marrow in approximately 50% of patients require a blood transfusion. Compounds of General formula I do not have these disadvantages. Their antiviral activity is not associated with cytotoxicity in the ical preparations can also be used in combination with other drugs for treatment and prophylaxis of the abovementioned diseases. Examples of these other drugs include tools that can be used for the treatment and prevention of HIV infections or diseases that accompany this disease, such as 3'-azido-3'- deoxythymidine, 2',3'-dideoxynucleoside, such as 2',3'- dideoxycytidine, 2', 3'-dideoxyadenosine and 2',3'- dideoxyinosine, acyclic nucleosides (e.g. Acyclovir), non-nucleoside RT inhibitors, protease inhibitors, such as, for example, invirase, interferons such as, for example, interferon, cytokines and interleukins (e.g. interleukin 16), chemokines, such as, for example, MIP1 , MIP1 , CCl, inhibitors of the renal secretions, such as, for example, probenecid, an inhibitor of the transport of nucleosides, such as, for example, dipyridamole, as well as immunomodulators such as interleukin II or stimulating factors, such as, for example, factors, colony stimulating granulocyte-macrophage (GM-CSF), factors, colony stimulating granulocyte (G-CSF, metropoitan), thrombopoetin and thrombopoietin-like factors. Compounds according to the present invention and the other drug can be used each separately, possibly at the same time, respectively, in a single dosage form or in two different lechenich salts of compounds of General formula I can be treated primarily salts of alkali and alkaline earth metals and ammonium carboxyl and phosphonate groups. As the alkali metal salts are preferred salts are lithium, sodium and potassium. As salts of alkaline earth metals can be considered, in particular, salts of magnesium and calcium. Under the ammonium salts according to the invention refers to salts containing ammonium ion, which can be substituted by up to four alkyl residues with 1 to 4 carbon atoms and/or Uralkalij residues, preferably benzyl residues. The substituents can be identical or different.

Under the esters of carboxylic acids of the lipid derivatives phosphonocarboxylate acids refers to a pharmacologically acceptable esters, preferably esters derived from benzyl, choline, ethanolamine, carnitine, C5-C7-cycloalkyl or a linear or branched alkyl residue containing 1-6 C-atoms of carbon, in particular, stands, ethyl, propylene, bootrom, Pentium, hexyl, isopropyl, ISO-bootrom, tert-bootrom, neopentyl or Taxila. Very particularly preferred methyl, ethyl, propyl, butyl, tert-butyl and benzyl.

Esters lepidopterology acids in vitro have the same efficiency as the corresponding carbon is ture carboxylic acid compounds of the formula I are subject to less degradation in the acidic environment in the decarboxylation and, consequently, the provide associated with this is the best bioavailability. In accordance with this prescribed dose can be significantly reduced in comparison with the free carboxylic acid. In addition, improving permeability through the membrane, for example, overcoming the blood-brain barrier and the passage through the cell membrane into the target cell. As an ester of the carboxylic acid must be cleaved by esterases only in vivo, the half-life in serum is lengthened.

In the General formula I, the residue R1preferably denotes a linear C10-C12is an alkyl group,

R1in particular, is a decile, undecyloxy, dodecyloxy or tridecanol group

n preferably denotes one of the numbers 0 or 1,

R2preferably denotes a linear C9-C122-alkyl group,

R2in particular, is a decile, undecyloxy or dodecyloxy group.

Preferred paired phosphonocarboxylate acids, respectively, their esters in the proposed conjugates of General formula I are the following acids and their esters:

- postemergence acid

- phosphono what you include n=0 and the combination of R1decyl/R2= dodecyl, R1=undecyl/R2=undecyl or R1=dodecyl/R2=decyl and, in addition, R1=undecyl/R2=decyl, R1=tridecyl/R2=decyl, R1=dodecyl/R2=undecyl.

Compounds of General formula I can be obtained because:

1) compound of General formula II

< / BR>
in which R1, R2and n have the above meanings, is subjected to the interaction with the compound of General formula III

< / BR>
in which m has the above meaning and R represents one of the above esters, in the presence of, if necessary substituted acid chloride arylsulfonic acid in an organic basis, respectively, in the presence of a base in an inert organic solvent, and optionally then transferred ester carboxylic acid alkaline saponification in a derived formula I, respectively in its physiologically acceptable salt, or

2) get mixed anhydride of the compound of formula III and the acid chloride alkyl - or arylsulfonic acid and injected it into reaction with the alcohol of formula II in the presence of a base in an inert organic solvent, respectively, directly at the base, and in the case neophocaena acid of the formula III, in which R denotes hydrogen, with an alcohol of the formula II in the presence of a base and, if necessary substituted acid chloride arylsulfonic acid and optionally transferred to a physiologically acceptable salt, or

4) the mixed anhydride of the compound of formula III in which R denotes hydrogen, and the acid chloride alkyl - or arylsulfonic acid is injected into reaction with the alcohol of formula II in the presence of a base, respectively, in an inert organic solvent, and if necessary, the conjugate was transferred to a physiologically acceptable salt, or

5) dichlorohydrin phosphonic acids of General formula IV

< / BR>
which can be obtained according Bangle al. (Synthetic Commun., 17, 1071 (1987)), based on bis-trimethylsilyl ester phosphonic acid, by subsequent interaction with oxalylamino, is subjected to interaction with alcohol of General formula II in the presence of a base in a molar ratio of 1:1, or

6) the compound of formula III is transferred using oxalicacid, as described in Tetrahedron Letters 33, 7473 (1992), in the appropriate dichlorohydrin phosphonic acid of the formula IV, which is then subjected to interaction with alcohol of formula II in the presence of a base in a molar ratio of 1:1. Prodat in ester carboxylic acids by alkaline saponification in a derived formula I, accordingly, in its physiologically acceptable salt.

Free acid lipid derivatives phosphonocarboxylate acids can be optionally converted into the corresponding esters.

Obtaining compounds of General formula II are described in the examples in EP 0545966.

Containing the compounds of formula I of the medicinal product for the treatment of, for example, viral infections may be applied in liquid or solid form interline or parenteral. An acceptable common forms of application, such as tablets, capsules, pills, syrups, solutions or suspensions. As injection medium preferably use water containing conventional in injection solutions of additives, such as stabilizers, agents dilution buffer and mix. Such additives are, for instance, tartrate and citrate buffer, ethanol, complexing agents such as ethylenediaminetetraacetic acid and its non-toxic salts, high molecular weight polymers such as liquid polyethylene oxide to regulate viscosity.

Liquid carriers for injection solutions must be sterile and preferably, they poured into ampoules. Solid carriers are, for example, starch, lactose is you, such as stearic acid, gelatin, agar-agar, calcium phosphate, magnesium stearate, animal and vegetable fats, solid high molecular weight polymers, such as polyethylene glycols, etc. Preparations suitable for oral administration, optionally, may contain flavorings and sweeteners.

The compounds of formula I fundamentally can be administered to the patient orally, intratrahealno, rectal, nasal, vaginal, sublingual, intravenously, intraarterially, intramuscularly, intradermally or subcutaneously. The dosage may depend on various factors such as method of application, type of disease, age and individual condition of the patient. Compounds according to the invention is usually used in an amount of from 0.1 to 1000 mg, preferably from 2 to 800 mg, especially preferably from 30 mg to 250 mg per day and kg body weight. Daily dose is preferably divided 2-5 techniques, and at each visit take 1-2 tablets containing the active substance from 0.5 to 3000 mg. Tablets can also be retardirani, so the number of doses per day is reduced to 1-3. The content of the active substance in retardirovannah tablets can range from 20 to 5000 mg of Active VESDA 10000 mg per day.

In addition to the compounds mentioned in the examples and the resulting combination of all mentioned in the claims of the values of the substituents in the scope of the present invention can be considered, the following compounds of formula I:

1. (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovaniya acid

2. (3-dodecyloxyphenyl-2-decyloxy)propoxy - fosforilirovaniya acid

3. (3-dodecylsulfonate-2-decyloxy)propoxy - fosforilirovaniya acid

4. (3-undecylenate-2-decyloxy)propoxy - fosforilirovaniya acid

5. (3-decelerate-2-decyloxy)propoxy - fosforilirovaniya acid

6. (3-tridecylamine-2-decyloxy)propoxy - fosforilirovaniya acid

7. (3-undecylenate-2-undecyloxy)propoxy - fosforilirovaniya acid

8. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovaniya acid

9. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovaniya acid

10. (3-dodecylmercaptan-2-undecyloxy)propoxy - fosforilirovaniya acid

11. (3-decelerate-2-dodecyloxy)propoxy - fosforilirovaniya acid

12. (3 undecylenate - fosforilirovaniya acid

14. (3-dodecylmercaptan-2-nonyloxy)propoxy - fosforilirovaniya acid

15. (3-undecylenate-2-nonyloxy)propoxy - fosforilirovaniya acid

16. (3-dodecylmercaptan-2-octyloxy)propoxy - fosforilirovaniya acid

17. (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovanija acid

18. (3-dodecyloxyphenyl-2-decyloxy)propoxy - fosforilirovanija acid

19. (3-dodecylsulfonate-2-decyloxy)propoxy - fosforilirovanija acid

20. (3-undecylenate-2-decyloxy)propoxy - fosforilirovanija acid

21. (3-decelerate-2-decyloxy)propoxy - fosforilirovanija acid

22. (3-tridecylamine-2-decyloxy)propoxy - fosforilirovanija acid

23. (3-undecylenate-2-undecyloxy)propoxy - fosforilirovanija acid

24. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovanija acid

25. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovanija acid

26. (3-dodecylmercaptan-2-undecyloxy)propoxy - fosforilirovanija acid

27. (3-decelerate-2-dodecyloxy)propoxy - phosphenelectrostimulation (3-dodecylmercaptan-2-dodecyloxy)propoxy - fosforilirovanija acid

30. (3-dodecylmercaptan-2-nonyloxy)propoxy - fosforilirovanija acid

31. (3-undecylenate-2-nonyloxy)propoxy - fosforilirovanija acid

32. (3-dodecylmercaptan-2-octyloxy)propoxy - fosforilirovanija acid

33. (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovaniya acid

34. (3-dodecyloxyphenyl-2-decyloxy)propoxy - fosforilirovaniya acid

35. (3-dodecylsulfonate-2-decyloxy)propoxy - fosforilirovaniya acid

36. (3-undecylenate-2-decyloxy)propoxy - fosforilirovaniya acid

37. (3-decelerate-2-decyloxy)propoxy - fosforilirovaniya acid

38. (3-tridecylamine-2-decyloxy)propoxy - fosforilirovaniya acid

39. (3-undecylenate-2-undecyloxy)propoxy - fosforilirovaniya acid

40. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovaniya acid

41. (3-undecylenoyl-2-undecyloxy)propoxy - fosforilirovaniya acid

42. (3-dodecylmercaptan-2-undecyloxy)propoxy - fosforilirovaniya acid

43. (3-decelerate-2-dodecyloxy)propoxy - fosforilirovaniya acid

44. (3 undecyloxy - fosforilirovaniya acid

46. (3-dodecylmercaptan-2-nonyloxy)propoxy - fosforilirovaniya acid

47. (3-undecylenate-2-nonyloxy)propoxy - fosforilirovaniya acid

48. (3-dodecylmercaptan-2-octyloxy)propoxy - fosforilirovaniya acid

49. methyl ether (3-dodecylmercaptan-2-decyloxy)propoxy - phosphenelectrostimulation acid

50. methyl ether (3-undecylenate-2-decyloxy)propoxy - phosphenelectrostimulation acid

51. methyl ether (3-tridecylamine-2-decyloxy)propoxy - phosphenelectrostimulation acid

52. methyl ether (3-undecylenate-2-undecyloxy)propoxy - phosphenelectrostimulation acid

53. methyl ether (3-dodecylmercaptan-2-undecyloxy)propoxy - phosphenelectrostimulation acid

54. methyl ether (3-decelerate-2-dodecyloxy)propoxy - phosphenelectrostimulation acid

55. methyl ether (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovanii acid

56. methyl ether (3-undecylenate-2-decyloxy)propoxy - fosforilirovanii acid

57. methyl ether (3-tridecylamine-2-decyloxy)propoxy - fosforilirovanii acid

58. methyl ether (3-undecylenate-2-undeci is poxi - fosforilirovanii acid

60. methyl ether (3-decelerate-2-dodecyloxy)propoxy - fosforilirovanii acid

61. ethyl ester (3-dodecylmercaptan-2-decyloxy)propoxy - phosphenelectrostimulation acid

62. ethyl ester (3-undecylenate-2-decyloxy)propoxy - phosphenelectrostimulation acid

63. ethyl ester (3-tridecylamine-2-decyloxy)propoxy - phosphenelectrostimulation acid

64. ethyl ester (3-undecylenate-2-undecyloxy)propoxy - phosphenelectrostimulation acid

65. ethyl ester (3-dodecylmercaptan-2-undecyloxy)propoxy - phosphenelectrostimulation acid

66. ethyl ester (3-decelerate-2-dodecyloxy)propoxy - phosphenelectrostimulation acid

67. ethyl ester (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovanii acid

68. ethyl ester (3-undecylenate-2-decyloxy)propoxy - fosforilirovanii acid

69. ethyl ester (3-tridecylamine-2-decyloxy)propoxy - fosforilirovanii acid

70. ethyl ester (3-undecylenate-2-undecyloxy)propoxy - fosforilirovanii acid

71. ethyl ester (3-dodecylmercaptan-2-undecyloxy)propoxy - fosforilirovanii acid

72. this is the fir (3-dodecylmercaptan-2-decyloxy)- propoxy-phosphenelectrostimulation acid

74. isopropyl ether (3-undecylenate-2-decyloxy)- propoxy-phosphenelectrostimulation acid

75. isopropyl ether (3-tridecylamine-2-decyloxy)- propoxy-phosphenelectrostimulation acid

76. isopropyl ether (3-undecylenate-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

77. isopropyl ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

78. isopropyl ether (3-decelerate-2-dodecyloxy)- propoxy-phosphenelectrostimulation acid

79. isopropyl ether (3-dodecylmercaptan-2-decyloxy)- propoxy-fosforilirovanii acid

80. isopropyl ether (3-undecylenate-2-decyloxy)- propoxy-fosforilirovanii acid

81. isopropyl ether (3-tridecylamine-2-decyloxy)- propoxy-fosforilirovanii acid

82. isopropyl ether (3-undecylenate-2-undecyloxy)- propoxy-fosforilirovanii acid

83. isopropyl ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-fosforilirovanii acid

84. isopropyl ether (3-decelerate-2-dodecyloxy)- propoxy-fosforilirovanii acid

85. neopentylene ether (3-dodecylmercaptan-is yloxy)- propoxy-phosphenelectrostimulation acid

87. neopentylene ether (3-tridecylamine-2-decyloxy)- propoxy-phosphenelectrostimulation acid

88. neopentylene ether (3-undecylenate-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

89. neopentylene ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

90. neopentylene ether (3-decelerate-2-dodecyloxy)- propoxy-phosphenelectrostimulation acid

91. neopentylene ether (3-dodecylmercaptan-2-decyloxy)- propoxy-fosforilirovanii acid

92. neopentylene ether (3-undecylenate-2-decyloxy)- propoxy-fosforilirovanii acid

93. neopentylene ether (3-tridecylamine-2-decyloxy)- propoxy-fosforilirovanii acid

94. neopentylene ether (3-undecylenate-2-undecyloxy)- propoxy-fosforilirovanii acid

95. neopentylene ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-fosforilirovanii acid

96. neopentylene ether (3-decelerate-2-dodecyloxy)- propoxy-fosforilirovanii acid

97. benzyl ether (3-dodecylmercaptan-2-decyloxy)propoxy - phosphenelectrostimulation acid

98. benzyl ether (3-undecylenate-2-decyl what epoxy - phosphenelectrostimulation acid

100. benzyl ether (3-undecylenate-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

101. benzyl ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-phosphenelectrostimulation acid

102. benzyl ether (3-decelerate-2-dodecyloxy)propoxy - phosphenelectrostimulation acid

103. benzyl ether (3-dodecylmercaptan-2-decyloxy)propoxy - fosforilirovanii acid

104. benzyl ether (3-undecylenate-2-decyloxy)propoxy - fosforilirovanii acid

105. benzyl ether (3-tridecylamine-2-decyloxy)propoxy - fosforilirovanii acid

106. benzyl ether (3-undecylenate-2-undecyloxy)- propoxy-fosforilirovanii acid

107. benzyl ether (3-dodecylmercaptan-2-undecyloxy)- propoxy-fosforilirovanii acid

108. benzyl ether (3-decelerate-2-dodecyloxy)propoxy - fosforilirovanii acid

Example 1

The disodium salt of R,S-(3-dodecylmercaptan-2-decyloxy)propoxy - phosphenelectrostimulation acid (DMDOP-PFA) and methyl ester (DMDOP-PFA-OMe)

18.2 ml complex trimethylboron ether phosphonopropionic acid are dissolved in 140 ml of dichloromethane and mixed at peremeshivanija dissolved in methanol and the solution every time again evaporated. The residue is dissolved in 30 ml of absolute pyridine and mixed with a solution of 48.7 R,S-(3 - dodecylmercaptan-2-decyloxy)-propan-1-ol. The mixture is evaporated to dryness, the residue under stirring mix from 47.1 g of 2,4,6-tri - isopropylbenzenesulfonyl and 150 ml of absolute pyridine. First, a thick suspension in approximately 30 min becomes thinner, and it is stirred for 25 h at room temperature.

The precipitate is sucked off and washed with a small amount of pyridine. The filtrate is mixed with 150 ml of water with stirring, the mixture stirred for 30 min at room temperature, evaporated and mixed with air. Again precipitated precipitate is filtered off and the ethereal filtrate is shaken with 0.5 N. HCl. The ether phase is well washed with water, dried and evaporated.

The remainder (84,2 g) purified by chromatography on silica gel in the solvent system dichloromethane/methanol/glacial acetic acid (9:0,5:0,5). Containing the product fractions evaporated. Get to 45.4 g of the corresponding complex ester of R,S-(3-dodecylmercaptan-2 - decyloxy) propoxy-phosphenelectrostimulation acid (DMDOP - PFA-OMe).

Thin layer chromatography on silica gel:

Rf= 0,3 (ether acetic acid/acetone/glacial acetic acid/water (10: slots 5 g of the product obtained above are dissolved in 70 ml of tetrahydrofuran and mixed with a 6.7 ml of 2 N. NaOH. Stirred for 4 h and left overnight. The reaction mixture through the buffer 2 - ethylhexanoic acid adjusted to pH 8 and evaporated. The residue is stirred with acetone and precipitated precipitated product is sucked off. Obtain 4.1 g of the acid with TPL. 242-246oC (decomposition).

Thin layer chromatography on silica gel:

Rf=0,31 (isopropanol/butyl acetate/water/conc.ammonia 10:6:3:1);

13C-NMR in D2O:COOH (d, 175 M. D., Jp-c-231,4 Hz)

Example 2

The disodium salt of R, S-(3-dodecylmercaptan-2-decyloxy) propoxy-fosforilirovanii acid (DMDOP-PAA) and methyl ester (DMDOP-PAA-OMe)

Analogously to example 1, on the basis of complex methyl ester phosphonooxy acid in the form of a waxy product, and (3-dodecyl-mercapto-2-decyloxy)-propan-1-ol, receive specified in the header of the connection with TPL=358-360oC (decomposition).

DMDOP-PAA:

Thin layer chromatography on silica gel:

Rf= 0,53 (n-butanol/glacial acetic acid/water 2:1:1);

Rf= 0,07 (dichloromethane/glacial acetic acid/water 9:0,5:0,5)

DMDOP-PAA-OMe:

Thin layer chromatography on silica gel:

Rfor = 0.6 (n-butanol/glacial acetic acid/water 2:1:1)

Rf= 0,1 (itro (CFU-GM Assay=test for granulocyte-macrophage colony-forming unit)

Conducted tests on GM-CFU, as described by Seidel and Kreja (Seidel H. und J. Kreja, L. , Blut 47, 139-145, 1983). Bone marrow cells (1105cells/ml) in Balb/c mice were cultured in Iscove medium, which contains 0.8% of methylcellulose, 20% horse serum, 10-4M - diglycerin and the optimal amount (12.5 or 25 ml) mouse serum, activated by endotoxin, which was obtained from Balb/c mice 4 h after intravenous injection of 50 μg of endotoxin per animal (Salmonella abortus equi; Sigma, Deisenhofen, Germany). After 6-day incubation for 24 h, colonies were stained with 2 (p-itfeel)-3-(p-nitrophenyl)-5-phenyl-tetrasilicate hydrate (INT, Sigma) and then counted using automatic image processor (Artek 982, Biosys GmbH, Karben, Germany).

In table. 1 shows the IC50-concentration of several conducted by the concentration dependent experiments with phosphonopropionic acid, DMDOP-PFA, phosphonooxy acid, DMDOP-PAA complex ethyl ester (3-octadecyl-hydroxy-2-hydroxy) - propoxy-phosphenelectrostimulation acid (OOHP-PFAE) and (3 octadecylamine-2-hydroxy)-propositionalization acid (OOHP-PFA) in comparison with the cytostatic agent cisplatin (Cis-DDP) and doxorubicin. As follows from the table. 1, DMDOP-PFA and DMDOP-PAA up to high power the on/macrophage series. Although this also applies to postemergency acid, phosphonooxy acid and conjugates OOHP-PFAE and OOHP-PFA is more toxic than DMDOP-PFA and DMDOP - PAA.

Example 4

Determination of oral bioavailability in a murine model of cytomegaly virus (MCMV)

Immunocompromising females mouse Balbus/e were injected intraperitoneal dose 8105plaque-forming units (PFU). Survival of animals was increased in a series of untreated<treated with foscarnet<DMDOP-PFA
Appendix 1

1. Determination of the activity depending on structure: Sn3-the options

Foscarnet-conjugates, i.e. aviaprovoda in Sn3component (exception: HDP 99.0008, see table. 2), were tested in vivo for their antiviral activity in animal models of mousepox cytomegaly (MCMV).

1.1 Methods

The MCMV model described in Appendix A.

1.2 Results

The data presented in the table. 3, illustrate the effectiveness of the compounds tested on the model of MCMV. The results show the percentage of surviving animals (30 days or more) in this system acute viral infection. All values expressed as% of the surviving rats compared with the control group with simplerow the derivative has been very active in vivo in animal models of mousepox cytomegaly after intraperitoneal administration,

suddenly found a clear correlation between the structure of the compound and its activity. Antiviral activity of the tested compounds to a large extent depends on the length of alkyl chain foscarnet,

suddenly most of these efiromaslichnyh also active after oral administration,

unexpectedly oral bioavailability also depends on the length of alkyl chain foscarnet,

"unexpectedly, in contrast to the HDP 99.0005 derivatives of acids HDP 99.0008 show only moderate activity after intraperitoneal administration and are completely inactive after oral administration.

Appendix a

Testing of conjugates of afrolatino and foscarnet model in vivo murine virus cytomegaly (MCMV)

Various conjugates of afrolatino and foscarnet were investigated in vivo in a model of murine virus cytomegaly. In this system, the degree of survival after infection with a virus murine cytomegaly was determined after 30 or more days after infection compared to the control treated with placebo.

Animals were infected (except for control animals) intraperitoneally 2105doses PFU per animal on day 0. All animals except the control b which has been created compounds were injected once daily intraperitoneally or orally in the amount of 30 mgkg-1day-1(0 days +1 h after infection) through day +8. The number of surviving animals was determined after 30 days or more.

Annex 2

2.1 the Relationship structure - activity: lengths Sn1-chain conjugates esters and foscarnet, i.e. derivatives with different length Sn1- chain (table. 4) were tested for antiviral activity in animal models of MCMV in vivo.

2.1.1 Methods

The MCMV model described in Appendix A.

2.1.2 Results

The data presented in the table. 5, illustrate the effectiveness of the compounds tested on the model of MCMV. The results represent the percentage of surviving animals (30 days or more) in this system acute viral infection. All values expressed as% of the surviving rats compared with the control group with simulated treatment. The data presented in table 5, indicate a clear relationship between structure and activity, depending on the length Sn1-alkyl side chain.

Annex 3

Antiviral activity of conjugates of foscarnet on animal models of mousepox cytomegaly (MCMV) after intraperitoneal administration: a comparison of various compounds

3.1 Methods

Connection t is the chemical structure of compounds tested in this series of experiments.

Table. 7 illustrates the effectiveness of various compounds (HDP 99.0046, HDP 99.0047, HDP 99.0015, HDP 99.0016) compared to HDP 99.0005, HDP 99.0008 and HDP 99.0040.

Shown:

all compounds tested in vivo antiviral activity, are not active and/or toxic,

- opposite the connection HDP 99.0005, HDP 99.0008 and HDP 99.0040 are unusually active in animal models of MCMV. In particular, the comparison of results for HDP 99.0016 compounds and compounds HDP 99.0040 showed that thioester derived HDP 99.0040 is unusually active in contrast to O-alkyl analogue.

Annex 4

4. Determination of the toxicity of compounds HDP 99.0005 compared with Hostetler connection HDP 99.0015 intravenously female mouse NMRI: lost/total number of animals in the group.

4.1 Method

The female mouse NMRI aged 6-8 weeks were treated intravenously a single dose on day 0. Blood parameters were determined on a blood analyzer on day +1 and +4. Animals were killed on day +4.

4.2 Results

In table. 8 shows the tolerance depending on the dose of the compounds HDP 99.0005 and HDP 99.0015 after intravenous administration to female mice NMRI. In producenci effect depending on the dose connection HDP 99.0015 with a 100% fatal after 10 min after intravenous injection of 100 mg/kg of this compound. In contrast, the connection HDP 99.0005 was well tolerated by all animals in the highest tested dose, i.e., 150 mg/kg

Such toxicity depending on the dose HDP 99.0015 was observed already at the minimum level of the test dose, i.e. at 25 mg/kg, the number of white blood cells through one day after the introduction, the data are given in table. 9.

1. The phosphonolipids carboxylic acids of General formula I

< / BR>
where R1denotes a linear alkyl (C9- C13;

R2denotes a linear alkyl (C8- C12;

R3denotes hydrogen, linear alkyl, C1- C6in particular, methyl, ethyl, propyl, butyl, pentyl, hexyl or benzyl;

n = 0, 1, or 2;

m = 0 - 3,

their physiologically acceptable salts with inorganic and organic bases.

2. The compounds of formula I under item 1, in which R1means decile, undecyloxy or tridecanol group.

3. The compounds of formula I according to one of p. 1 or 2, in which R2means decile, undecyloxy or dodecyloxy group.

4. The compounds of formula I according to one of paragraphs.1 to 3, in which n = 0 or 1.

5. The compounds of formula I according to one of paragraphs.1 to 4, in which m = 0, 1, or 2.

what do benzyl.

7. Means exhibiting antiviral activity and contains the active substance is a compound of formula I on PP.1 - 6 and pharmaceutical excipients and carriers.

Priority on the basis of R3- the hydrogen atom from 15.12.1995.

Priority on the basis of R3- alkyl or benzyl from 22.10.1996.

 

Same patents:

The invention relates to 1H-indol-3-acetamide General formula I where X is oxygen; R1selected from groups (i), (iii), where (i)6-C20-alkyl, C4-C12-cycloalkyl; (iii) - (CH2)n-(R80), where n is 1-8 and R80is the group specified in (i); R2is hydrogen, halogen, C1-C3-alkyl, C1-C2-alkylthio,1-C2-alkoxy; R3each independently is hydrogen or methyl; R4- R7each independently - C1-C10-alkyl, C2-C10alkenyl,3-C8-cycloalkyl,1-C10-alkoxy,

WITH4-C8-cycloalkane, phenoxy, halogen, hydroxy, carboxyl, -C(O)O(C1-C10-alkyl), hydrazide, hydrazino, NH2, NO2, -C(O)NR82R83where R82and R83independently is hydrogen, C1-C10-alkyl or a group of formula (a), where R84and R85independently is hydrogen, C1-C10-alkyl; p= 1 to 5; z is a bond, -O-, -NH-; Q is-CON(R82R83), -SO3H, phenyl, a group of formula b), C), (d), where R86independently selected from hydrogen, C1-C10-alkyl, and their pharmaceutically acceptable salts or their esters, or Amida

The invention relates to a method for producing phosphonic compounds of the formula I

< / BR>
by reaction of phosphite formula II

< / BR>
with a compound containing a carbon-carbon double bond, of the formula III

X-CH=CH-Y

where R1and R2independently of one another denote phenyl or C1-C4-alkyl which may be substituted by one or more chlorine atoms or bromine, X represents hydrogen or methyl and Y represents-COOR1, -CONH2, -CONHR1-CONR21or-CN, where R1above

The invention relates to the chemistry of organophosphorus compounds, namely to a new method of obtaining S-dialkyl-, alkylphenyl and diphenylarsinic esters of 4-methoxyphenylacetone acids of General formula I

< / BR>
where

Ar = 4-MeOC6H4;

R is lower alkyl, phenyl;

R' and R ' = lower alkyl, phenyl

The invention relates to new tizamidine pyridinylmethyl acids f-ly R2-Z-Q-(CR1R1)n-CH[P(O)(OH)2]2(I) where R1-H, -SH, -(CH2)mSH or-S-C(O)-R3, R3- C1-C8-alkyl, m = 1 - 6, n = 0 to 6, Q is a covalent bond or-NH-, Z - pyridinyl, R2- H, -SH, -(CH2)mSH, -(CH2)mS-C(O)R3or-NH-C(O)-R4-SH, where R3and m have the above meaning, R4- C1-C8-alkylen, or their pharmaceutically acceptable salts or esters

The invention relates to polypeptides, used as immunological reagents for the identification, prevention, and treatment of infections caused by HCV

The invention relates to biochemistry, namely to obtain biologically active substances with antiviral activity

The invention relates to medicine, in particular to pharmacology relates to antiviral agents in the form of solid dosage forms containing acyclovir, microcrystalline cellulose, salt of stearic acid, low molecular weight polyvinylpyrrolidone, and, optionally, a dye in a certain ratio, and method of its production
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