DE4103904A1 - NEW GAMMA PYRONS AND THEIR USE AS MEDICINAL PRODUCTS AND METHOD FOR THE PRODUCTION THEREOF - Google Patents

Abstract

The invention relates to novel gamma-pyrones, gamma-pyridones and gamma-thiopyrones and their use as medicaments against viral diseases, preferably for the treatment of retroviral diseases, comprising compounds of the formula (I), in which R1 and R2 stand for hydrogen, a branched-chain or unbranched-chain C1 to C26, preferably C1 to C6, alkyl, C1 to C26 alkenyl or C1 to C26 alkinyl group or combinations thereof; R3 stands for a branched-chain or unbranched-chain C1 to C26 alkyl group; and R4 stands for a C3 to C26 cycloalkyl, branched-chain or unbranched-chain alkyl, alkenyl or alkinyl group. The hydrogen atoms of the methylenic group in R1, R2 and R4 may be substituted by O alkyl (C1 to C6), O acyl, O aryl, O aralkyl and alkyl (C1 to C6) groups, and in R4 also by halogenated groups, oxo groups and hydroxylic groups with double links. The elements represented by Z are independently from each other O, S or NH.

Description

The invention relates to new gamma pyrones and their Use as a medicine to treat viral Diseases.

Spread viral diseases such as hepatitis B and -C or AIDS exponentially. Therefore, the development has been antiviral Active ingredients are of great economic importance acquired and is at the center of pharmacological Research. The current antiviral drugs are characterized by a low antiviral spectrum of activity and in most cases by a high one Toxicity. There are serious disadvantages poor applicability and rapid induction more resistant Virus strains in appearance.

As antiviral prophylactics, amantadine and Rimantadine for some influenza viruses and metisazone against smallpox viruses. Amantadine and Rimantadine have low toxicity. As another antiviral are idoxuridines, trifluorothymidine, ethyl deoxyuridine and iododeoxycytidine, which are particularly common in some local herpes infections only externally can be applied. However, these substances are sometimes highly toxic.

Adenine arabinoside and cytosine arabinoside are pronounced Immunosuppressive and very side effects. they cause, for example, changes in the chromosomes.  

Bromovinyl deoxyuridine works well against herpes simplex Type 1 virus and some other viruses, but not against Herpes simplex virus type 2. Acyclovir also works against some herpes viruses, but not against the cyto megalovirus. Dihydroxyproposymethylguanine works against this Epstein-Barr virus and the cytomegalovirus, however, is very toxic.

Phosphonoformate, suramin and zidovudine work among others against HIV viruses, but have a high toxicity.

Many of the above substances work against one viral thymidine kinase, a viral polymerase or a viral reverse transcriptase. For many of the substances is already developing resistance during therapy of the infecting virus has been observed.

Recently, beneficial substances have proven to be beneficial Proven to be polyanions such as dextran sulfate and pentosan polysulfate, because they are specifically against reverse transcriptase of retroviruses, especially HIV, seem to work. DE-OS 36 01 136 describes the use of organic Polymers, the inorganic anionic Groups included for prophylaxis and therapy of retroviral Infections in mammals.

EP-OS 02 93 826 describes therapeutic and prophylactic Effects of sulfated polysaccharides against AIDS.

The technical problem of the invention is one antiviral agent with low toxicity, high specific Efficacy, good applicability and if possible  low tendency to induce resistant virus strains to provide.

This problem is caused by gamma pyrones as new substances of formula (I)

solved, wherein R₁ and R₂ H, a branched or unbranched C₁ to C₂₆ alkyl, preferably C₁ to C₆, C₁ to C₂₆ alkenyl or C₁ to C₂₆ alkynyl group or combinations these are, R₃ is a branched or unbranched C₁ to C₆ is alkyl group and R₄ is a C₃ to C₂₆ cycloalkyl, branched or straight chain alkyl, alkenyl or Is alkynyl group, and the hydrogen atoms of the methylene groups in R₁, R₂ and R₄ can be replaced by O Alkyl (C₁ to C₆) -, O acyl, O aryl, O-aralkyl and Alkyl (C₁ to C₆) groups, and in R₄ additionally by Halogen groups, double bonded oxo groups and hydroxyl groups and Z each independently = O, S or NH, and when R₁ = H, R₂ ≠ H and when R₂ = H, R₁ ≠ is H and the following compounds are excluded are

and the following connections are also excluded are  

The object is further achieved by containing pharmaceuticals the compound of formula (I), wherein R₁ to R₄ and Z have the same meaning as previously described.

In a particular embodiment, as in the drug containing compounds of the formula (II)

used, wherein R₁, R₂, R₅, X and Y have the following meaning to have  

These compounds can be used to make drugs for the treatment of viral diseases, preferred of retroviral diseases used in mammals will.  

Gamma-pyrone derivatives are available from Dehy dracic acid over 3,5-dioxo-hexanoic acid methyl ester. The Dehydroacetic acid is substituted by appropriately Acetoacetic acid derivatives formed. From dehydroacetic acid becomes a corresponding by reaction with magnesium methoxide substituted 3,5-dioxo-hexanoic acid methyl ester won, which can still be derivatized, for example by alkylation at CH-acidic positions. This is followed by a recycling step. Preferably is used as a DBU reagent for cyclization. This leads to modifications in the side chains the corresponding structure types (F. Wangemann, dissertation Berlin 1989, "Synthesis of gamma pyrones from Podelepis hieracioides).

Gamma-pyrone structures without alkoxy functions are obtained by condensation of carboxylic acids or their derivatives (A. N. Sagredos et. Al. Liebigs Ann. Chem. 706, 90 to 94, volume 697, pages 111 to 115). From Canadian Journal of Chemistry, 56, 1796-1799 (1978) become Methods for the alkylation of pyronones, the starting compounds for the structures claimed according to the invention are described.

From Tetrahedron lett. 2167 (1976) and for example J. Org. Chem. 43, 4966, 1878 is the representation of thiopyran 4-thio derivatives described. G. Voss and H. Describe Gerlach in Liebigs Ann. Chem., 1982, pages 1466 to 1477, the synthesis of nitrogen analogs Pyrones by reacting the corresponding pyrones with Ammonia in methanol by heating in a sealed Glass tube.  

The preparation of the compounds of formula (I) can also by isolating compound (II) from species the Australian podolepsis plant, particularly from Podolepsis longipedata A. and from the roots of Podolepsis rugata and from Podolepis hierakioides. The substances are obtained by extraction in organic solvents and subsequent cleaning and processing of the Extract by means of chromatography. The corresponding Methods are described in F. Bohlmann et. al., Phytochemistry, Vol. 26, pages 187 to 190, 1987 and J. Jakupovic et al. Phytochemistry, vol. 28, pages 3497 to 3501, 1989.

The substances thus obtained were tested in a test system checked for their antiviral effect. As a test system served the determination of virus replication in HIV-infected Blood cells. The system allows antivirals Detect substances associated with virus replication and interfere with the infection potential. Such substances could penetrate virus particles into the Prevent cell interior and those in the already infected Virus-propagated cells could not create new target cells find more. That way would be an effective one Prophylaxis against antiviral diseases possible and it Therapy could be initiated to prevent intrusion prevented by viruses in the cell interior. The consequence is that the infection stops and that therapeutic goal is achieved quickly and easily. Other intracellular targets for antiviral agents are conceivable, for example the so-called Uncoating or removing infectious virions.

The antiviral test system is designed to be used for the model virus (HIV) all antiviral substances displays without prior knowledge of their  to have a molecular target or mechanism of action. The strength of the test system lies in its sensitivity, Specificity and breadth of possible destinations for antiviral Substances. The test system shows protease inhibitors also to the RT inhibitors or blockers of virus uptake through the host cell. Also across virus classes Active substances, such as polyanions with their broad The spectrum of activity is displayed safely. The high sensitivity the use of non-transformed, human peripheral blood cells guaranteed.

The system is particularly suitable for spraying new ones antiviral substance classes with so-called lead structures, d. H. a basic structural motif, its chemical Derivatives the pharmacological, here antiviral, Affect properties. The lead structures are then Basis for a targeted synthesis of these derivatives.

The gamma pyrones according to the invention fell in this test due to their antiviral effectiveness. With a concentration of 10 µmol, which is less than the tenth part of the corresponds to half-maximum cytotoxic concentration, was four days after human HIV exposure Lymphocyte decreased HIV replication was observed. This is surprisingly a new antiviral Substance class has been discovered that the gamma-pyrone structure as a common structural characteristics.

Embodiment

The gamma pyrones of Table 1 were as from Australian Podolepis herakioides obtained by extraction and then purified by chromatography.  

Table 1

Table 1 shows the substances obtained and their antiviral activity measured as follows:  

A substance-induced inhibition of HIV multiplication in Cell culture is determined by the quantitative determination of a viral structural protein (p24) and viral nucleic acid (HIV-RNA) determined. For the virus multiplication one finds Culture of human lymphocyte application.

The substance to be tested is after a 10 mM solution by serial 1:10 dilutions in the culture medium to one in the previous cytotoxicity test approximate non-toxic concentration set. Human lymphocytes are made from donor blood isolated. For this purpose, the "buffy coats" of the preserved blood the lymphocytes by means of density gradient centrifugation won.

In a safety laboratory that complies with the L3 guidelines cells with human immunodeficiency Virus (HIV) infected. The infected cells are then at 37 ° C and a water vapor saturated Atmosphere containing 5% CO₂ in the presence of those to be tested Incubated substance for four days.

The concentration of HIV-specific core protein p24, that after infection of the lymphocytes by the virus producing Cells are secreted into the culture supernatant, is determined using a "Sandwich Elisa" test. The evaluation is done photometrically. From one Calibration curve and the absorption values obtained, which the Cultures of HIV-infected lymphocytes result in The presence of the examined dance has grown, the concentration of protein p24 is calculated. The substance induced Inhibition of p24 production is caused by Comparison with an untreated control culture as a percentage calculated.  

The HIV RNA formed in the infected lymphocytes becomes detected by nucleic acid hybridization. The evaluation takes place photometrically. From a calibration curve and the absorption values obtained, which the cultures HIV-infected lymphocytes result in the presence the substance to be examined is washed the concentration of HIV-RNA is calculated. The substance-induced Inhibition of HIV RNA production is by comparison with an untreated control culture as a percentage calculated.

The viability of the infected cells is checked using the Trypan blue method determined. Infected cells that are in Presence of the substances to be examined for four days cultured are mixed with a trypan blue solution. In a microscopic analysis is done by counting the proportion of blue-colored, dead cells is determined. The vitality is determined from the difference between total cells and dead cells and is also in percent specified.

Fig. 1 shows the dose-response relationship for inhibition of protein 24, inhibition of HIV-RNA and viability for use 4 from Table 1.

Claims (6)

1. Gamma-pyrones of the formula (I) wherein R₁ and R₂ are hydrogen, a branched or unbranched C₁ to C₂₆ alkyl, preferably C₁ to C₆, C₁ to C₂₆ alkenyl or C₁ to C₂₆ alkynyl group or combinations thereof, R₃ is a branched or unbranched C₁ to C₆ alkyl group and R₄ is a C₃ to C₂₆ cycloalkyl, branched or straight-chain alkyl, alkenyl or alkynyl group, and the hydrogen atoms of the methylene groups in R₁, R₂ and R₄ can be replaced by O alkyl (C₁ to C₆) -, O acyl, O aryl, O -Aralkyl and alkyl (C₁ to C₆) groups, and in R₄ additionally by halogen groups, double-bonded oxo groups and hydroxyl groups and Z are each independently of one another = O, S or NH, where when R₁ = hydrogen, R₂ ≠ is hydrogen and when R₂ = Hydrogen, R₁ ≠ is hydrogen and the following compounds are excluded: if and the following connections are also excluded: 2. Medicament containing the compound of the formula (I), where R₁ to R₄ and X have the same meaning as in claim 1. 3. Medicament according to claim 2, characterized in that the following compounds are used as compound (I) where R₁, R₂, R₅, X and Y have the following meaning 4. Process for the preparation of a compound of formula (I), characterized in that the compounds (II), wherein R₁, R₂, R₅, X and Y have the same meaning have as in claim 3, from the plant species Podolepsis be isolated and by conventional methods be derivatized.   5. Use of compounds of formula (I) for the preparation a drug used to treat viral Diseases. 6. Use according to claim 5 for the treatment of retroviral Diseases.