WO1998020013A1 - New knoevenagel condensation products, method for their production and their use - Google Patents

Abstract

The present invention relates to new Knoevenagel condensation products, the method for their production and their use, specially for pharmaceutical and analytical purposes. Further, conjugates of Knoevenagel condensates with biomolecules are disclosed. The invention is characterized by the compound of formula (3).

Description

New Knoevenagel condensation products, process for their production and use

description

The present invention relates to new Knoevenagel condensation products, processes for their production and their use, in particular for pharmaceutical and analytical purposes. Furthermore, conjugates of the Knoevenagel condensates with biomolecules are disclosed.

A publication by Feigl (Z. Anal. Chem. 74 (1 928), 380-386) describes the use of p-dimethylaminobenzylidene rhodanine for the detection of silver. Escobar Godoy and Guiraum Perez (Analyst 1 1 1 (1 986), 1 297-1 299) describe the use of the pyridoxal analog (Z) - pyridoxylidene rhodanine for the spectroscopic quantification of silver.

The problem underlying the present invention was to provide new compounds which can be used in particular for pharmaceutical and analytical purposes. This object is achieved by the synthesis of the compound (Z) -5'-O-phosphonopyridoxidenidhhodanin (3), which is a derivative of the vitamin B6 coenzyme pyridoxal-5'-phosphate (1). Compound (3) is prepared by reacting (1) with the synthetic heterocyclic compound rhodanine (2) in a Knoevenagel condensation (cf. also Kesel et al., Tetrahedron 52 (1 8.1 1 .1 996), 14787-14800).

Compound 3 is named (Z) -5 - [[5-hydroxy-6-methyl-3- [(phosphonooxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4-thiazolidinone. The rhodanine portion of this molecule is able to bind to biomolecules such as single-stranded nucleic acids. The charged phosphate group ensures good water solubility. Compound 3 is subject to a (Z / E) stereoisomerism to form the (E) isomer 4. This cis / trans rearrangement is triggered, for example, by UV radiation with the wavelengths 254 nm and / or 366 nm.

In dipolar aprotic solvents the originally yellow compound 3 is present as red 3-pyridinolate 5 in the form of a free acid, e.g. in pyridine or dimethyl sulfoxide (DMSO).

Pyridoxal-5'-phosphate rhodanine (Z) -5'-0-phosphoruso 'pyridoxylidei

The yellow compound 3 forms a deep red monosodium salt, the structure of which could be elucidated by X-ray structure analysis. It was crystallized as hemiheptadecahydrate (8.5 hydrate) and is present as (Z) - stereoisomer 6. When exposed to UV radiation, the monosodium salt 6 is also subject to the cis / trans rearrangement to the red (E) stereoisomer 7.

Derivatives of compound 3 can be obtained by reaction with C ₁₂ -C ₁₈ fatty acid chlorides on the OH group.

The present invention further relates to compounds of the general formula (8).

wherein: X ¹ , X ² and X ^{3 are} each independently selected from O, S and NR ⁵ ;

Is YO or S; Z is CR ⁵ or N;

R \ R ² , R ³ , R ⁴ and R ^{5 are} each independently selected from any substituent; R ^{6 is} selected from optionally substituted hydrocarbon radicals and O and n is 0 or 1; and salts, for example salts with alkali metal, alkaline earth metal or ammonium ions, hydrates and stereoisomers thereof, with the proviso that the compound is not pyridoxylidene rhodanine.

In the compounds of the general formula (8), X ¹ is preferably O. X ² is preferably NR ⁵ , where R ^{5 is} hydrogen or CC ₄ alkyl and particularly preferably hydrogen. X ³ is preferably S. Y is preferably S. Z is preferably CR ⁵ , where R ^{5 is} hydrogen or C _r C ₄ alkyl and particularly preferably hydrogen.

REPLACEMENT BIAΓΓ (RΓ.GFL 26) The substituents R ¹ to R ⁴ can in themselves be any substituents provided that they are compatible with the overall structure. Optionally, R ¹ and R ² or / and R ³ and R ⁴ can be bridged. Examples of the substituents R ¹ to R ⁴ are hydrogen, halogen, hydroxyl, amine and optionally substituted alkyl radicals, for example C to C ₆ alkyl, aminoalkyl, hydroxyalkyl, alkoxy etc. Particularly preferably, at least one of the substituents R to R ⁴ contains a sub physiologically negatively charged group, for example an acid group such as phosphate, carboxylate, sulfonate etc.

The compounds of the general formula (8) are particularly preferably the (Z) -5-O-phosphonopyridoxylidene rhodanine (3), and salts, hydrates and stereoisomers thereof.

The present invention further provides a process for the preparation of a compound of the general formula (8), characterized in that a compound of the formula (9)

reacted in a Knoevenagel condensation with a compound of the formula (1 0)

REPLACEMENT BUTT (RULE 26) wherein X ¹ , X ² , X ³ , Y, Z, R ¹ -R ⁵ and R ^{6 are} as defined for the compounds 8. The reaction is preferably carried out in an essentially equimolar ratio of 9 to 10 in a suitable solvent or solvent mixture. The temperature is preferably from room temperature to the reflux temperature of the solvent.

The compounds of general formula (8) surprisingly have pharmaceutical activity. The invention thus furthermore relates to a pharmaceutical composition which contains as active ingredient a compound as stated above (preferably without taking into account the disclaimer in claim 1) and, if appropriate, pharmaceutically customary additives, auxiliaries, carriers and diluents.

A first important application of the compounds according to the invention is the treatment of infectious diseases, for example viral infections, parasitic diseases, fungal diseases or bacterial diseases. Particularly preferred is the use for the treatment of viral infections, in particular those viral infections which are caused by enveloped viruses, such as hepadnaviruses, herpes viruses or retroviruses, e.g. HIV. However, the compounds according to the invention have also proven suitable for the treatment of diseases which are caused by other viruses, such as, for example, influenza or papilloma viruses. Furthermore, the compounds according to the invention have proven to be very suitable for controlling parasites such as, for example, Leishmania, Plasmodia or Trypanosomes. The compounds are most preferably used in HIV therapy, for example in combination therapy with other agents such as AZT or DDI.

In addition, the compounds according to the invention are also used to combat tumor diseases, for example leukemia, in particular T-cell leukemia, or skin tumors such as malignant

Melanoma or Kaposi's sarcoma. Surprisingly, they show a selective cytotoxic effect on malignant cells, which does not occur in normal cells.

The compounds 8 also have immunomodulatory effects and are therefore suitable, for example, for the treatment of autoimmune diseases such as, for example, multiple sclerosis, Alzheimer's disease, lupus erythematosus, myasthenia gravis, chronic arthritis, type I diabetes, etc. and various neurodegenerative diseases.

Furthermore, the compounds 8 are suitable for the treatment of diseases which exist in connection with disorders of the vitamin B metabolism, e.g. as a vitamin B6 antagonist.

Diseases which are related to disorders of enzymatic reactions are yet another application. The compounds of the invention act as effectors, e.g. Activators, inhibitors or modifiers, on enzymatic reactions of the caspase system, and in particular as inhibitors of nitrogen monoxide synthases.

In addition, it was found that compounds 8 can influence the secretion of TNFσ in T lymphocytes. They are also able to bind to the cellular CD38 receptor, thereby inhibiting the binding of HIV gp41. In this way, the signal transmission path is decoupled from CD38, which leads to an inhibition of syncytia formation, NO release, T cell death by apoptosis, and single cell lysis.

The compounds according to the invention can be used as such for therapeutic purposes. However, they can also be used as chelates with polyvalent metal ions or as noncovalent and / or covalent conjugates with biomolecules, for example nucleic acids, proteins, lipids, fatty acids etc. The pharmaceutical compositions according to the invention can be in any form, for example as creams, ointments, injectable or orally administrable liquids, liposomal formulations, tablets, dragées, capsules etc. The application can be, for example, locally or systemically, topically, orally or by injection. The dosage can be varied within a wide range, for example from 0.01 μg to 1 mg per kg of body weight, depending on the type of disease and application.

In addition, the compounds according to the invention are also suitable as detection reagents. On the one hand, they can be used in the form of radioactive, for example with ³ H, ¹⁴ C, ³² P or ³⁵ S-labeled derivatives. However, use in unlabelled form is preferred, since its strong coloring or fluorescent properties make it easy to detect in biological systems. In particular, the compounds according to the invention are therefore suitable for the detection of biomolecules, for example in diagnostic analysis methods, for example for the detection of single-stranded nucleic acids. The compounds are also suitable for structure elucidation, for example for sequencing proteins or nucleic acids.

In addition, the compounds according to the invention can also be used for electrochromic applications, e.g. can be used as molecular digital switch substances or indicators for pH value, temperature and solvent.

Furthermore, the compounds or derivatives thereof according to the invention are suitable as absorbers for ions, organic substances etc. or due to their intense colors and reversible color changes for decorative applications. In this context, particular mention should be made of the alkaline earth metal salts, for example Mg or Ca salts of the compounds, which are in the form of gels. Yet another area of application is nanotechnology, where, for example, individual fluorescent dye molecules and biomolecule derivatives excited with laser systems are detected and detected by sensitive detection systems, for example CCD cameras or avalanche detectors and optical techniques on thin-layer plates, in membranes, cells, drops, capillaries, etc. can be located. Further possibilities for single-molecule localization are optical scanning near-field microscopy on mini chips, confocal microscopy in polyacrylamide gels, for example 2-dimensional or multidimensional gel electophoresis of nucleic acids.

Finally, the present invention also relates to derivatives of the compounds (8) which result from hydrolytic degradation and have, for example, the general formulas 11, 12 or 13:

ERSA1ZBUTT (RULE 26)

wherein X ¹ , X ² , X ³ , Y, Z, R ¹ -R ⁴ , R ⁵ and R ^{6 are} as defined for the compounds 8.

The invention is further illustrated by the following examples and illustrations. Show it:

Fig. 1: the protection of cells against HIV infection mediated by compound (3).

Fig.2: a schematic representation of hydrolysis by-products that arise during the synthesis of compound (3);

Examples

Manufacturing process of 3

Production of the raw product:

3.83 g of 2-thioxo-4-thiazolidinone (rhodanine) (M = 133, 18 g / mol; n = 28.76 mmol) are dissolved in 150 ml of absolute ethanol in a 50 ml round-bottomed flask with a reflux condenser, and 7.63 g of pyridoxal-5'-phosphate monohydrate (M = 265.16 g / mol; n = 28.78 mmol). The pyridoxal 5'-phosphate monohydrate almost does not dissolve in the cold.

REPLACEMENT BUTT (RULE 26) The mixture is boiled on a water bath with stirring under reflux. The pyridoxal-5'-phosphate monohydrate now slowly goes into solution and an initially yellow, then orange, then red suspension is formed. After boiling for 20 minutes, 1 50 ml of water are added through the cooler because the suspension begins to belch. The mixture is heated for a further 10 min, during which the red reaction product precipitates and leads to belching. Then refrigerate in the freezer at a temperature of - 18 ° C for no more than 2 hours.

Processing of the raw product

The orange product is filtered off, transferred with the mother liquor and with 100 ml of ice-cold abs. Washed ethanol. Vacuum drying in a desiccator over anhydrous calcium chloride; Yield: 9.85 g of orange-yellow crude product (Z) -5 - [[5-hydroxy-6-methyl-3- [phosphonooxy) methyl] -4-pyridinyl] - methylene] -2-thioxo-4-thiazolidinone. The product contains educts, by-products and, despite vacuum drying, ethanol and water.

Purification of the raw product by ion exchange reprecipitation:

9.85 g of crude product are suspended in 610 ml of water in a 2000 ml round bottom flask. 200 ml of a solution [β (NaHCO ₃ ) = 100 mg / ml] of anhydrous sodium hydrogen carbonate [n (NaHCO ₃ ) = 238, 10 mmol] saturated at room temperature (22 ° C.) are added in several portions. This now red solution is titrated with 238, 10 ml (238, 10 mmol HCl) of 1, 0 M hydrochloric acid. The resulting mixture is stirred for a further 15 minutes. At the end of the titration (pH 2) the color changes from red to yellow and the acid 3 fails. It is suctioned off and dried for 24 hours at a pressure of 4 mbar and a temperature of 70 ° C. Then it is dried for 24 h at a pressure of 0.001 mbar in an oil pump vacuum at room temperature. Yield: 8.70 g (83%, based on the total synthesis) of yellow, amorphous powder (Z) -5 - [[5-hydroxy-6-methyl-3- [(phosphonooxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4-thiazolidinone, content w> 96% (1H-NMR).

The analysis of compound 3 comprises ¹ H-NMR spectroscopy, ¹³ C-NMR spectroscopy, ³¹ P-NMR spectroscopy, RP-18 HPLC, fast atom bombardment mass spectrometry (FAB MS), IR spectroscopy, UV spectrophotometry and fluorescence spectrophotometry. Typical analytical data of compound 3 follow.

C _1l H ₁₁ N ₂ O ₆ PS _2, M = 362.31 g / mol. F.198 ° C - 201 ° C (with decomposition, uncorrected).

FABMS (MH ⁺ calculated for m / z 362.99): m / z (τe \. Int.) 363.0 (12.0%, MH ⁺ ), 227.4 (18.1%), 270.4 (8.7%), 171.2 (8.3%), 282.4 (7.1%), 267.1 (2.8%), 283.4 (2.1%), 265.1 (1.5%).

¹ H NMR (TFA-cZ): "52.91 (s, 3 H, 2'-CH ₃ ), 5.42 (d, | ³ ( ³¹ P, ¹ H) | = 8.0 Hz, 2 H, 5'-CH ₂ -OPO ₃ H ₂ ), 7.76 (s, 1 H, 4'-CH), 8.47 (s, 1 H, 6-CH). ¹ H NMR (DMSO-tf6): δ 2.42 (s, 3 H, 2'-CH ₃ ), 4.89 (d, | ³ ( ³¹ P, ¹ H) | = 8.0 Hz, 2 H, 5'-CH ₂ -OPO ₃ H ₂ ), 7.55 (s, 1 H, 4'-CH), 7.84 (s, 1 H, 6-CH). 1HNMR (pyridine-c / ₅ ): δ 2.72 (s, 3 H, 2'-CH ₃ ), 5.62 (d, | ³ J ( ³¹ P, ¹ H) | = 7.5 Hz, 2 H, 5'CH ₂ -OPO ₃ H ₂ ), 8.36 (s, 1 H, 4'CH), 8.53 (s, 1 H, 6-CH). ¹³ C NMR, BB-decoupled (TFA-c /): δ 16.83 (s, 2'-CH ₃ ), 65.75 (s, 5'-CH ₂ - OPO ₃ H ₂ ), 121.04, 122.13 (2 s, C -4 '), 133.34 (s, C-6), 136.85, 136.91 (2 s, RR'C-SR "), 138.54 (s, C-5), 142.05 (s, C-4), 147.64 (s, C-2), 153.03 (s, C-3), 171.93 (s, C = O), 193.39 (s, C = S). ³¹ P NMR (TFA-tY): rf-4.24 (s, R-OPO ₃ H ₂ ).

FT IR (KBr): 3430 (v NH, m), 1713 (v HN-C = O, s), 1213 (v P = O in /? O (HO) PO ₂ -, s), 1046 (vC = SinS-CS-N, s), 1024 (v P = O in /? O (HO) PO ₂ ^" , s). UV / VIS (H ₂ O): Λ _maXιl = 232 nm W (1% / 1 cm ) = 373], Λ _{max, 2} = 308 nm [A (1% / 1 cm) = 271], Λ _{max, 3} = 353 nm [A (1% / 1 cm) = 413], λ _maxA = 454 nm [A (1% / 1 cm) = 227]. US / VIS (MeOH): λ _maXι1 = 291 nm [A (1% / 1 cm) = 245], λ _maXι2 = 347 nm

REPLACEMENT BUTT (RULE 26) 11a

[A (1% / 1 cm) = 489].

UV / VIS (DMSO): Λ _max = 518 nm [A (1% / 1 cm) = 98].

Fluorescence excitation spectrum (ex) and emission spectrum (em) (DMSO):

Λ _em = 575 nm: Λ _{ex max ,,} = 355 m, Λ _{ex, max, 2} = 397 nm, λ _{e ma 3} = 451 nm, λ _{eXι max 4} = 467 nm, Λ _{ex, maX 5} = 483 nm, λ _{eXι max, 6} = 493 nm, Λ _{ex, max, 7} = 518 nm; Λ _ex - 490nm: Λ _{em, max} = 575 nm.

Fluorescence excitation spectrum (ex) and emission spectrum (em) (pyridine):

Λ _em = 575 nm: Λ _{ex, max ,,} = 451 nm, Λ _{ex, max, 2} = 467 nm, λ _{e max, 3} = 471 nm, Λ _βχ , max, 4 = 483 nm, λ _{eXι max} , ₅ = 493 nm, Λ _ex , _max , ₆ = 518 nm; λ _ex = 490nm: Λ _{em, max} - 575 nm.

REPLACEMENT BUTT (RULE 26) - 1 2 - 2. Manufacturing process of 6

2.1

A mass of 363 mg (Z) -5 - [[5-hydroxy-6-methyl-3 - [(phosphonooxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4-thiazolidinone (3) ( 1.0 mmol) is mixed with 20.0 ml of 0.10 M sodium hydroxide solution (2.00 mmol of NaOH). After mixing with 10.0 ml abs. Ethanol the solution is cooled for 24 h at a temperature of -1 8 ° C. The fine crystalline needles are suctioned off. Yield: 340 mg (63%) red (Z) -5 - [[3-hydroxy-2-methyl-5- [(phosphonooxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4-thiazolidinone monosodium salt -hemiheptadeca (8 1/2) hydrate (6). Before an elemental analysis, this product was dried in a water gun at a pressure of 0.1 mbar in a water jet pump vacuum at a temperature of 70 ° C. for two days: deep red (Z) -5 - [[5-hydroxy-6-methyl-3- [ (phosphonoxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4-thiazoIidinone monosodium salt hemipenta (2 1/2) hydrate (6).

2.2

8.70 g of dry compound 3 (24.00 mmol) are mixed with 240.00 ml of a 0.1 M sodium hydroxide solution (24.00 mmol NAOH) and then mixed with 1 24.00 ml of ethanol. The mixture is kept at a temperature of -1 8 ° C for 24 h. The precipitated red crystals are filtered. 7.55 g (73%) of compound 6 are obtained.

F.205 ° C-208 ° C (with decomposition, uncorrected).

Calculated for CH ₁₀ N ₂ Na O ₆ PS ₂ o 2 1/2 H ₂ O, M = 429.33 g / mol; C 30.77% H 3.52% N 6.52%, found C 30.97% H 3.43% N 6.35%. FT IF (KBr): 3277 (v OH water, m, broad), 1 700 (v HN-C = O, w), 1 229 (v P = O in RP (HO) PO ₂ \ s), 1 1 98 (v HN-C = S, s), 1,090 (v C = S in S-CS-N, s), 973 (v POC in PO-CH2-aryl, 2). - 1 3 - 3. Detection method for single-stranded nucleic acids

Compounds 3 and 6 can be used in radiolabelled form, for example labeled with ³ H, ¹⁴ C, ³² P or ³⁵ S, as probes for the detection of single-stranded nucleic acids.

The substances 3/6 can also be used for the fluorescence detection of single-stranded DNA. The (E) stereoisomer 4 fluoresces in DSMO, the (Z) stereoisomer in water significantly less. When the (E) stereoisomer 4 is bound to single-stranded DNA, single-stranded DNA can be detected when fluorescence occurs after extraction with DMSO. The wavelength used for excitation is in the range from 420 to 500 nm, the emission in DMSO or pyridine is 575 nm.

Compounds 3, 4 and 6 show a noncovalent association with single-stranded DNA. This non-covalent bond can be fixed photochemically by irradiation of the UV light of the wavelengths 254 and / or 366 nm. 3 and 4 react with a thymine nucleobase methyl group photochemically with the elimination of H ₂ S.

14

4. Inhibition of isoforms of the enzyme nitric oxide synthase

Substance 6 inhibits the neuronal, endothelial and immunologically / - inducible isoforms of nitrogen monoxide synthase (NOS). The inhibition constant (IC ₅₀ ) for nNOS is 61.25 µM. For eNOS and iNOS, the inhibition constant (IC ₅₀ ) is 50μM. These values are determined by measuring the conversion of ( ³ H) arginine to ( ³ H) citrulline using recombinant human NOS isoforms isolated from CHO cells.

REPLACEMENT BUTR (RULE 26) - 1 5 - 5. Two-dimensional nucleic acid electrophoresis

Substance 3 can be used in radioactively labeled or unlabelled form for the two-dimensional electrophoretic separation of single-stranded nucleic acids. In the first dimension there can be a separation according to the charge and in the second dimension a separation according to the molecular size. Compound 3 can be associatively or covalently coupled to the nucleic acid.

6. Therapeutic application

Substance 3 and derivatives thereof are effective as therapeutic agents, in particular for influencing the propagation processes of infectious agents, e.g. from those that occur in the host with intermediates as single-stranded DNA, such as retroviruses, herpes viruses or hepatoviruses.

6.1 HIV inhibition

Myeloid HUT78 cells are exposed to HIV infection in the absence or in the presence of 1, 87 mM or 0, 1 87 mM compound 3. The HIV detection is carried out by photometric determination of the p24 antigen. From Figure 1 it can be seen that no p24 was detectable in the cells treated according to the invention, whereas in the control the p24 test was positive from the seventh day.

6.2 Treatment of herpes

One patient (1 7 years, male) with labial herpes is treated with one

Olive oil-water emulsion of compound 3 (about 1 mg / ml) treated. - 1 6 -

Total remission is found a few hours after local application to the affected skin.

6.3 Treatment of influenza

A patient (51 years, male) with virus flu is orally treated with a few drops of an H ₂ O / DMSO solution of compound 3 (approx. 1 μg / ml). The disease symptoms disappeared overnight.

7. Temperature / solvent indicator

The (Z / E) stereoisomerism of compound 3 occurs depending on the temperature (thermochromism) and depending on the solvent (solvatochromism). 3 can therefore be used as a temperature indicator and / or solvent indicator.

8. Digital switching function

The (Z / E) stereoisomerism of compounds 3 and 4 occurs depending on the application of an electric field (electrochromism). Therefore 3 can be used as a digital switching element due to the different fluorescence spectroscopic properties compared to 4 e.g. when excited by laser light with a wavelength of 480 nm. Thereby, after applying an electric field to 3, rearrangement in 4 occurs and thus fluorescence emission up to 575 nm after excitation at 480 nm.

9. Vitamin B6 antagonism

Compound 3 can act in metabolism as an antagonist of the coenzyme pyridoxal-5'-phosphate by binding to the enzymes which require this coenzyme and therefore have tumor-inhibiting properties. - 1 7 -

10. Proliferation control

Compound 3 can be used for the proliferation control / chemotherapy / cytostatic therapy of malignant tumors in local application to skin tumors (malignant melanoma, Karposi sarcoma), possibly in combination with UV radiation, and / or for the proliferation control of other skin diseases with an increased cell division rate (e.g. Psoriaris) can be used. The binding of compound 3 to replication-active single-stranded DNA regions can be photochemically fixed by UV radiation and leads to locally limited cytotoxicity.

1 1. Structure elucidation

Compound 3, due to its coloring and fluorescent properties, as well as its ability to couple to biomolecules, e.g. Proteins are used for structure elucidation, e.g. for amino acid sequence determination of proteins or for DNA sequencing.

12. Detection of hydrolysis by-products

The hydrolysis by-products that result from the synthesis of compound 3 are examined by mass spectroscopy.

The corresponding connections are listed in Fig. 2.

Claims

18th

Expectations

Compounds of the general formula (8)

wherein:

X ¹ , X ² and X ^{3 are} each independently selected from O, S, and NR ⁵ ;

Y is O or S;

Z is CR ⁵ or N;

R ¹ , R ² , R ³ , R ⁴ and R ^{5 are} each independently selected from any substituent;

R ^{6 is} selected from optionally substituted hydrocarbon radicals and O and n is 0 or 1; and salts, hydrates and stereoisomers thereof, with the proviso that the compound is not pyridoxylidene rhodanine.

2. Compounds according to claim 1, wherein X ^{1 is} 0.

REPLACEMENT BUTT (RULE 26) - 1 9 -

3. Compounds according to any one of claims 1 or 2, wherein X ^{2 is} NR ⁵ .

4. Compounds according to any one of the preceding claims, wherein X ³ S is.

5. Compounds according to any one of the preceding claims, wherein Y is S.

6. Compounds according to any one of the preceding claims, wherein Z is CR ⁵ .

7. Compounds according to any one of the preceding claims, wherein one of the substituents R ¹ -R ^{4 contains} a negatively charged group.

8. Compounds according to one of the preceding claims, selected from (Z) -5 [[5-hydroxy-6-methyl-3 - [(phosphonooxy) methyl] -4-pyridinyl] methylene] -2-thioxo-4- thiazolidinone of formula (3)

and salts, hydrates and stereoisomers thereof. - 20 -

9. A process for the preparation of a compound of general formula (8), characterized in that a compound of formula (9)

reacted in a Knoevenagel condensation with a compound of the formula (10)

(10)

[il

wherein X ¹ , X ² , X ³ , Y, Z, R ¹ -R ⁵ and R ^{6 are} as defined in claim 1.

REPLACEMENT BUTT (REGFI 26) - 21 - 0. A process for the preparation of a compound according to claim 8, characterized in that pyridoxal-5'-phosphate is reacted with rhodanine in a Knoevenagel condensation.

1 . Pharmaceutical composition, characterized in that it contains, as active ingredient, a compound according to one of Claims 1 to 8 and, optionally, pharmaceutically customary additives, auxiliaries, carriers and diluents.

2. Use of a compound according to any one of claims 1 to 8 for the manufacture of a pharmaceutical composition for the treatment of infectious diseases, tumors and autoimmune diseases.

3. Use according to claim 1 2 for the preparation of an antiviral agent.

4. Use according to claim 1 3, for the preparation of an agent for the treatment of viral infections caused by hepatoviruses, herpes viruses or retroviruses.

5. Use of a compound according to any one of claims 1 to 8 for the manufacture of a pharmaceutical composition for the

Treatment of diseases related to disorders of the vitamin B metabolism.

6. Use of a compound according to any one of claims 1 to 8 for the manufacture of a pharmaceutical composition for the

Treatment of diseases related to disorders of the immune system. - 22 -

1 7. Use of a compound according to any one of claims 1 to 8 for the manufacture of a pharmaceutical composition for the treatment of diseases which are associated with disorders of enzymatic reactions.

1 8. Use according to claim 1 7 as an effector of the enzymatic activity of nitric oxide synthases.

1 9. Use of compounds according to any one of claims 1 to 8 as detection reagents.

20. Use according to claim 1 9 for the detection of biomolecules.

21. Use according to claim 20 for the detection of single-stranded nucleic acids.

22. Use of compounds according to one of claims 1 to 8 for electrochromic applications.

23. Conjugates of compounds according to one of claims 1 to 8 with biomolecules.

24. Derivatives of compounds according to one of the preceding claims of the general formulas (1 1), (1 2) or (1 3):

23 -

YH

R6

wherein X ¹ , X ² , X ³ , Y, Z, R ¹ -R ⁴ , R ⁵ and R ^{6 are} as defined in claim 1.

REPLACEMENT BUTT (RULE 26)