WO1993016091A1

WO1993016091A1 - New liponucleotides, their preparation and their use as antiviral medicaments

Info

Publication number: WO1993016091A1
Application number: PCT/EP1993/000294
Authority: WO
Inventors: Dieter Herrmann; Alfred Mertens; Harald Zilch
Original assignee: Boehringer Mannheim Gmbh
Priority date: 1992-02-12
Filing date: 1993-02-08
Publication date: 1993-08-19
Also published as: AU3453993A; DE4204032A1

Abstract

New phospholipid derivates of nucleosides having general formula (I) are disclosed. In the formula, R1 stands for a straight or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, which may if required be substituted one or several times by phenyl, halogen, C1-C6-alkoxy, C1-C6-alkylmercapto, C1-C6-alkoxycarbonyl, C1-C6-alkylsulfinyl or C1-C6-alkylsulfonyl groups; R2 stands for a straight or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, which may if required by substituted one or several times by phenyl, halogen, C1-C6-alkoxy, C1-C6-alkylmercapto, C1-C6-alkoxycarbonyl or C1-C6-alkylsulfonyl groups; X is a valency bond, oxygen, sulphur, sulfinyl or sulfonyl; Y has the same meaning as X, where both X and Y groups may be the same or different; Z can be oxygen or sulphur; A can be a methylene group or an oxygen atom; Nuc can be a residue derived from a nucleoside derivate. Also disclosed are the tautomers of these derivates and their physiologically tolerable salts of inorganic and organic acids or bases, as well as a process for preparing the same and medicaments containing these compounds, in particular for treating viral or retroviral infections.

Description

New liponucleotides, their production and their use as antiviral drugs

The present invention relates to new phospholipid derivatives of nucleosides of the general formula I,

CH2-Y-R2

in the

Rl is a straight-chain or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, optionally one or more times by phenyl, halogen, Ci-Cg-alkoxy, Cx-Cö-alkyl mercapto, Ci-Cg-alkoxycarbonyl -, Ci-Cg-alkylsulfinyl or Cj-Cs-alkylsulfonyl groups can be substituted,

2 a straight-chain or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, optionally one or more times by phenyl, halogen, Ci-Ce alkoxy, Ci-Cβ-alkyl mercapto, -C-C ₆ - alkoxycarbonyl - or Ci-Cg-alkylsulfonyl groups can be substituted, X represents a valence line, oxygen, sulfur, sulfinyl or sulfonyl,

Y has the same meaning as X, the two

Groups X and Y may be the same or different,

Z can be oxygen or sulfur,

A can represent a methylene group or an oxygen atom,

Nuc can be a residue derived from a nucleoside derivative, and

their tautomers and their physiologically tolerable salts of inorganic and organic acids or bases, as well as processes for their preparation and medicaments containing these compounds.

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

J. Biol. Chem. .265, 6112 (1990) and EP 0350 287 A2 describe the production and use of liponucleotides as antiviral drugs. However, only those known to nucleosides, such as e.g. AZT and ddC, coupled dimyristoylphosphatidyl and dipalmitoylphosphatidyl residues with their fatty acid ester structure.

J. Med. Chem. 3_3, 1380 (1990) describes nucleoside conjugates of thioether lipids with cytidine diphosphate which have an antitumor effect and could be used in oncology. Chem. Pharm. Bull. 1, 209 (1988) describes 5 '- (3-SN-phosphatidyl) nucleosides with antileukaemic activity and their enzymatic synthesis from the corresponding nucleosides and phosphocholines in the presence of phospholipase D with transferase activity .

The compounds of the present invention are new and also have valuable pharmacological properties. They are particularly suitable for the therapy and prophylaxis of infections caused by DNA viruses such as e.g. the herpes simplex virus, the cytomegalovirus, Papova viruses, the varicella zoster virus or Epstein-Barr virus or RNA viruses such as toga viruses or in particular retroviruses such as the oncoviruses HTLV-I and II, as well as the lentiviruses Visna and the human immunodeficiency virus HIV-1 and 2.

The compounds of the formula I appear to be particularly suitable for the treatment of the clinical manifestations of retroviral HIV infection in humans, such as persistent generalized lymphadenopathy (PGL), the advanced stage of the AIDS-related complex (ARC) and the clinical picture of AIDS.

Surprisingly, it has now been found that compounds of general formula I multiply DNA or RNA viruses in vivo, e.g. B. in the FVL model on the mouse, better inhibit than known liponucleotides. The inhibitory effect on the HI virus, the cause of the immune deficiency disease AIDS, is of particular therapeutic interest. Only 3'-azido-3 ^' deoxythymidine (DE-A-3608606) is currently approved for the treatment of AIDS in AIDS patients. However, toxic side effects of 3'-azido-3'-deoxythymidine on the bone marrow require blood transfusions in about 50% of the patients treated. The connections of all General Formula I do not have these disadvantages. They have an antiviral effect without being cytotoxic in pharmacologically relevant doses.

The compounds of the present invention and their pharmaceutical preparations can also be used in combination with other medicaments for the treatment and prophylaxis of the above-mentioned infections. Examples of these further medicaments include agents which can be used for the treatment and prophylaxis of HIV infections or diseases accompanying this disease, such as 3 'azido-3' deoxythymidine, 2 ', 3' dideoxynucleosides such as B. 2 ', 3'-dideoxycytidine, 2', 3 '-dideoxyadenosine and 2', 3 '-dideoxy-inosine, acyclic nucleosides (z. B. Acyclovir) or non-nucleoside RT inhibitors, such as. B. HEPT, nevirapine or L-697,661 and corresponding derivatives. The compounds of the present invention and the other medicament can each be administered individually, simultaneously, if appropriate in a single or two separate formulations or at different times.

Possible salts of the compounds of the general formula I are, in particular, alkali metal, alkaline earth metal and ammonium salts of the phosphate group. Lithium, sodium and potassium salts are preferred as alkali salts. Magnesium and calcium salts are particularly suitable as alkaline earth metal salts. According to the invention, ammonium salts are understood to be salts which contain the ammonium ion, which can be substituted up to four times by alkyl radicals having 1-4 carbon atoms and / or aralkyl radicals, preferably benzyl radicals. The substituents can be the same or different.

The compounds of the general formula I can contain basic groups, in particular amino groups, which can be converted into acid addition salts using suitable acids. Examples of suitable acids for this purpose are: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, fumaric acid, succinic acid, tartaric acid, citric acid, lactic acid, maleic acid or methanesulfonic acid.

In the general formula I, Rj preferably denotes a straight-chain Cg-C ^ alkyl group which can also be substituted by a C ^ -Cg alkoxy or a Ci-Cs-alkyl mercapto group. Ri represents in particular a decyl, undecyl, dodecyl, tridecyl or tetradecyl group. Preferred Ci-Cg-alkoxy substituents of Ri are the methoxy, ethoxy, butoxy and the hexyloxy groups. If ^ is substituted by a Ci-Cs-alkyl mercapto residue, this means in particular the methyl mercapto, ethyl mercapto, propyl mercapto, butyl mercapto and hexyl mercapto residues.

R2 preferably means a straight chain

group, which can also be substituted by a Ci-Cg-alkoxy group or a Cι ~ Cg-alkylraercapto group. R ₂ represents in particular a decyl, undecyl, dodecyl, tridecyl or tetradecyl group. The C 1 -C 6 -alkoxy substituents of R ₂ are preferably the methoxy, ethoxy, propoxy, butoxy and the hexyloxy group.

If R2 is substituted by a Ci-Cs-alkyl mercapto residue, this means in particular the methyl mercapto, ethyl mercapto, butyl mercapto and hexyl mercapto residue.

X and Y preferably represent an oxygen or sulfur atom, Z is preferably an oxygen atom.

Nuc is a nucleoside derivative represented by the 5 ^1-position to the phosphonic acid of the lipophilic part of the Formula I is bound. The following residues are suitable as nucleosides or nucleoside analogues:

in which

R _{3 is} hydrogen or a hydroxyl group,

₄ , R ₅ each represent hydrogen or one of the radicals 4 and R _{5 is} halogen, a hydroxyl, a cyano or an azido group and, moreover, R ₃ and R4 can represent a further bond between C-2 'and C-3',

B means one of the following compounds:

(IIIC)

(Illd) being

Rg can be hydrogen, an alkyl chain with 1-4 carbon atoms or halogen, R ₆ 'can be hydrogen or a benzyl or phenylthio radical,

R _{7 can be} hydrogen, an alkyl chain with 1-4 carbon atoms or halogen,

Rg can be hydrogen, an alkyl chain with 1-4 carbon atoms, halogen, or a hydroxy or an amino group,

Rg can be hydrogen or an amino group, and

Rio hydrogen, halogen, C -Cg-alkoxy, Ci-Cg-alkylmercapto, or an amino group which can be mono- or disubstituted by C-Cg-alkyl-, Ci-Cg-alkoxy-, hydroxy-C2 ~ Cg-alkyl - and / or Cs-Cg-cycloalkyl, aryl, hetaryl, aralkyl or hetarylalkyl groups, which may optionally be substituted in the aryl or hetaryl radical by one or more hydroxyl, methoxy or alkyl groups or halogen, or Allyl, which can optionally be substituted with mono- or dialkyl or alkoxy groups.

Nuc can also be of the carbocyclic type

B

or a cyclobutane, oxetanozine residue or a residue of the type -CH2-CH2-O-CH2-B or -CH ₂ -0-CH ₂ -CH2-B derived from seconucleoside derivatives, such as in WO90 / 09998 or WO90 / 09999, wherein R ₃ , R, R ₅ and B have the meanings given above.

In particular, such nucleosides or nucleoside analogues come into question for Nuc, which differ from the known antiviral acting connections, such as. B. carbovir, HEPT, gangciclovir, AZT or acyclovir.

In the nucleosides Nuc of the formula II, ₄ and R _{5 are} preferably each hydrogen or one of the two radicals is preferably cyano, azido or halogen, such as fluorine, chlorine, bromine or iodine.

Compounds in which R ₃ and R4 represent a hydrogen atom and R5 is cyano, azido or fluorine, or R5 is hydrogen and R ₃ / R _{4 represent} a further bond between C-2 'and C-3' are particularly preferred .

In the bases B of the formula III, R or R ₇ preferably denote a hydrogen atom, a methyl, ethyl, propyl or butyl radical, or a halogen atom, such as fluorine, chlorine, bromine or iodine. For _β or R7, a hydrogen atom, the methyl or ethyl radical and a chlorine or bromine atom are particularly preferred.

The radical R ₈ is preferably a hydrogen atom, a methyl, ethyl, propyl or butyl radical, an amino group or a halogen atom such as fluorine, chlorine, bromine or iodine, preferably chlorine or bromine.

Rio preferably means a hydrogen, fluorine, chlorine or bromine atom, a Ci-Cg-alkoxy group, in particular a methoxy-ethoxy, propoxy, butoxy or hexyloxy group, a C ~ Cg-alkyl mercapto group, in particular a methyl mercapto group , Ethylmercapto, butylmercapto or hexyl mercapto group, or an amino group which can be mono- or disubstituted by a C -Cg alkyl group, such as. B. the methyl, ethyl, butyl or hexyl group, by a hydroxy-C2-C ₆ alkyl group, such as. B. the hydroxyethyl, hydroxypropyl, hydrox - butyl or hydroxyhexyl group, by a C ₃ -C ₅ cycloalkyl rest, such as B. the cyclopropyl, cyclopentyl or cyclohexyl radical, preferably by aryl phenyl, by an aralkyl radical, such as in particular benzyl, which may also have one or more hydroxyl or methoxy groups, by Ci-Cg-alkyl groups, such as. B. the methyl, ethyl, propyl, butyl or hexyl group or by halogen atoms such as fluorine, chlorine or bromine may be substituted. The amino group can also be substituted by a heterarylalkyl or hetaryl radical, such as in particular z. B. the thienyl, furyl or pyridyl. The heterarylalkyl radical is preferably understood to mean the thienylmethyl, furylmethyl or pyridylmethyl radical.

Preferred coupled nucleosides in the claimed liponucleotides of the general formula I are:

3'dideoxy-3 'azidouridine

3 'dideoxyinosine

3 • -Dideoxyguanosine

3 • -Dideoxycytidine

3'-dideoxyadenosine

Deoxythy idin

3'-Dideoxy-2 ', 3'-didehydro-N ⁶ - (o-methylbenzyl) adenosine

3'-Dideoxy-2 ^• , 3'-didehydro-N ⁶ - (2-methylpropyl) adenosine

3'-dideoxy-3'-azidoguanosine

Deoxy-3 'azidothymidine

3 ^» -dideoxy-3'-fluoro-5-chlorouridine

^• deoxy-3 ^• fluorothymidine

3 • -Dideoxy-3'-fluoradenosine

3 • -Dideoxy-3'-fluoro-2,6-diaminopurine riboside

3'-dideoxy-2 •, 3'-didehydrocytidine

Deoxy-2 ^• , 3'-didehydrothymidine

The compounds of general formula I can be prepared by a compound of the general formula V,

R! -X-CH ₂

CH-A-P-OH (V),

II ¹ OH

R ₂ -Y-CH ₂

in which R, R, X, Y, Z and A have the meanings given, with a compound of the general formula VI,

Nuc - OH (VI)

in which Nuc has the meaning given above, preferably denotes a compound of the formula Via,

in which R3 'represents hydrogen or a hydroxy group protected by an oxygen protective group familiar to the person skilled in the art and R ₄ ' u. R5 'in each case represents hydrogen, halogen, an azido, a cyano or one of the radicals 4' and R5 'is a hydroxyl group protected by an oxygen protective group familiar to the person skilled in the art, or R3' and R4 'represent a further bond and B has the meanings given, in the presence of a condensing agent such. B. an optionally substituted benzenesulfonic acid chloride, preferably 2,4,6-triisopropylbenzenesulfonic acid chloride and a tert. Nitrogen base, e.g. As pyridine or lutidine, in an inert solvent such as. B. toluene, or reacted directly in pyridine and, after hydrolysis, optionally cleaves the oxygen-protecting groups in accordance with the methods customary in nucleoside chemistry, or

2. a compound of the general formula VII

R ¹ -X-CH ₂ CH ₃

Z.

CH- - IPI- -0-CH ₂ ~ CH ₂ -N-CH ₃ (VII)

R ² -Y-CH ₂ 0- CH ₃

in which R, R2, X, Y, Z and A have the abovementioned meanings, with a compound of the general formula VI or Via, in which R ₃ ', R4 •, Rs' and B have the meanings given, in Presence of phospholipase D in an inert solvent, such as. As chloroform, brings in the presence of a suitable buffer to the reaction and, if appropriate, splits off the oxygen protecting group after the reaction, in accordance with the methods customary in nucleoside chemistry.

The preparation of the compounds of the general formulas V and VII are described in DE-OS 39 29 217.7 and WO 91/05558. The compounds of general formula V with A = -CH2- are prepared from the corresponding halomethyl derivatives by reaction with trialkyl phosphite according to Michaelis-Arbusow under well-known conditions.

The phosphoric acid esters of the general formula V with A = 0 are prepared from the corresponding secondary alcohol by reaction with phosphoric acid amide chlorides or phosphoric acid ester chlorides and subsequent hydrolysis to give the free phosphoric acid monoesters, as described, for example, in J. Chem. Soc. 1963 (1958), Synthesis 32 (1982), Synthesis 737 (1977), J. Am. Chem. Soc. 71, 3822 (1949) or Nucl. Acid Chem., Ed. L.B. Townsend + R.S. Tipson, Wiley & Sons 1978.

The preparation of the compounds of general formula VI or Via are described for. B. in EP-A 0 286 028 and WO 90/08147.

Compounds similar to general formula I are described in EP-A-0350287. The corresponding 1,2-diesters of glycerol are described there.

The medicaments containing compounds of the formula I for the treatment of viral infections can be administered enterally or parenterally in liquid or solid form. The usual forms of application are possible, such as tablets, capsules, dragees, syrups, solutions or suspensions. Water is preferably used as the injection medium, which contains the additives customary for injection solutions, such as stabilizers, solubilizers and buffers. Such additives are, for example, tartrate and citrate buffers, ethanol, complexing agents, such as ethylene-dia-tetraacetic acid and their non-toxic salts, high-molecular polymers, such as liquid polyethylene oxide for crime regulation. Liquid carriers for injection solutions must be sterile and are preferably filled into ampoules. Solid carriers are, for example, starch, lactose, mannitol, methyl cellulose, talc, highly disperse silicic acids, higher molecular fatty acids, such as stearic acid, gelatin, agar-agar, calcium phosphate, magnesium stearate, animal and vegetable fats, solid high-molecular polymers, such as polyethylene glycols, etc. Preparations suitable for oral applications can, if desired, contain flavoring or sweetening agents.

The dosage can depend on various factors, such as the mode of application, species, age or individual condition. The compounds according to the invention are usually applied in amounts of 0.1-100 mg, preferably 0.2-80 mg per day and per kg of body weight. It is preferred to distribute the daily dose over 2-5 applications, 1-2 tablets with an active ingredient content of 0.5-500 mg being administered with each application. The tablets can also be delayed, which reduces the number of applications per day to 1-3. The active substance content of the retarded tablets can be 2 - 1000 mg. The active ingredient can also be given by continuous infusion, the amounts of 5-1000 mg per day normally being sufficient.

For the purposes of the present invention, in addition to the compounds mentioned in the examples and the combination of all the meanings of the substituents mentioned in the claims, the following compounds of the formula I are suitable:

1. (2 ', 3'-Dideoxy-3' -fluoro-5-chlorouridine) -5 • -phosphoric acid- (3-dodecylmercapto-l-decyloxy) -2-propyl ester

2. (3'-Deoxy-3'-azido-thymidine) -5 • -phosphoric acid- (3-dodecylsulfinyl-1-decyloxy) -2-propyl ester 3. (3'-Deoxy-3'-azido-thymidine) -5'-phosphoric acid- (3-dodecylsulfonyl-1-decyloxy) -2-propyl ester

4. (2 ', 3'-Dideoxycytidine) -5'-phosphoric acid- (3-dodecyl-mercapto-1-decyloxy) -2-propyl ester

5. (2 ', 3' -dideoxyinosine) -5'-phosphoric acid- (3-dodecyl-mercapto-1-decyloxy) -2-propyl ester

6. (2 ', 3'-Dideoxyguanosine) -5'-phosphoric acid- (3-dodecyloxy-1-decyloxy) -2-propyl ester

7. (2 ', 3'-Dideoxyadenosine) -5'-phosphoric acid- (3-dodecyl-mercapto-l-decyloxy) -2-propyl ester

8. (3'-Deoxythymidine) -5'-phosphoric acid- (3-dodecylmercapto-1-decylmercapto) -2-propyl ester

9. (3'-Deoxy-2 ', 3'-didehydrothymidine) -5'-phosphoric acid- (3-dodecylmercapto-l-decyloxy) -l-propyl ester

10. (3'-Deoxy-3'-fluorothymidine) -5'-phosphoric acid- (3-dodecylmercapto-1-decyloxy) -2-propyl ester

11. (2 ', 3' -Dideoxy-3 '-azidoguanosine) -5'-phosphoric acid- (3-dodecylmercapto-1-decyloxy) -2-propyl ester

12. (2 *, 3'-Dideoxy-3'-fluoro-2,6-diaminopurine riboside) -5'-phosphoric acid (3-dodecylmercapto-1-decyloxy) -2-proyl ester

13. [2 ', 3' -dideoxy-2 •, 3'-didehydro-N ⁶ - (2-ethylpropyl) adenosine] -5'-phosphoric acid- (3-dσdecylmercapto-1-decyloxy) -2- propyl ester 93/16091

-15-

14. [2 ', 3 • -dideoxy-2', 3'-didehydro-N ⁶ - (o-methylbenzyl) adenosine] -5'-phosphoric acid- (3-dodecylmercapto-l-decyloxy) -2- propyl ester

15. (2 ', 3'-Dideoxy-2', 3 '-dehydehydrocytidine) -5'-phosphoric acid- (3-decylmercapto-1-dodecyloxy) -2-propyl ester

16. (2 ', 3'-Dideoxy-3'-fluoroadenosine) -5'-phosphoric acid- (3-undecyloxy-1-dodecyloxy) -2-propyl ester

17. (2 ^" , 3'-Dideoxy-3'-azidouridine) -5'-phosphoric acid (3-decylsulfonyl-1-dodecyloxy) -2-propyl ester

18. (2 •, 3 ^• -dideoxycytidine) -5'-phosphoric acid- (3-decyl-mercapto-1-decyloxy) -2-propyl ester

19. (2 •, 3'-Dideoxyinosine) -5'-phosphoric acid- (3-dodecyl-mercapto-l-dodecyloxy) -2-propyl ester

20. (3'-Deoxy-3'-azidothymidine) -5'-phosphoric acid (3-tetra-decylmercapto-l-decyloxy) -2-propyl ester

21. (3'-Deoxy-3'-azidothymidine) -5'-phosphoric acid- (3-penta-decylmercapto-l-decyloxy) -2-propyl ester

22. (2 ', 3'-Dideoxyinosine) -5'-phosphoric acid- (3-tridecyl-mercapto-1-decyloxy) -2-propyl ester

23. (2 •, 3'-dideoxyinosine) -5'-phosphoric acid (3-dodecyl mercapto-l-octyloxy) -2-propyl ester

24. 3-Dodecylmercapto-2-decyloxymethyl-l-phosphonic acid 5'- (3'-deoxy-3'-azidothymidine) ester _25th ₃ - _U ndecylmercapto-2-undecyloxymethyl-l-phosphonic acid-5 '(3' -deoxy-3 ^'-fluorthy idin) ester

26. 3-Dodecylmercapto-2-dodecylmercaptomethyl-1-phosphonic acid 5 '- (3', 3 '-dideoxyinosine) ester

example 1

.3 '-Desoxy-3' -azidothymidine) -5'-phosphoric acid- (3-dodecyl-mercapto-1-decyloxy) -2-propyl ester

1.45 g (3 mmol) of phosphoric acid (3-dodecylmercapto-l-decyloxy) - 2-propyl ester and 800 mg (3 mmol) of AZT were extracted twice with 20 ml of abs. Pyridine added and evaporated. The residue was abs in 20 ml. Pyridine was added, 2.7 g (8.5 mmol) of 2,4,6-triisopropylbenzenesulfonyl chloride were added under nitrogen and the mixture was stirred for 24 hours at 40 ° C. Then 10 ml of water were added, the mixture was stirred for a further 2 hours at room temperature and the solvent was removed on a rotary evaporator .

The oily residue was freed of pyridine residues by evaporation with toluene and purified by column chromatography on silica gel 60 using a linear gradient from dichloromethane to dichloromethane / methanol 7.5 / 2.5 as the eluent. Yield 0.94 g (43% of theory), oil. Rf = 0.27 (CH ₂ Cl ₂ / MeOH 8/2), Rf = 0.63 (CH ₂ Cl2 / MeOH / H ₂ 0 6.5 / 2.5 / 0.4) on TLC plates Merck 5715 silica gel 60 F.

The phosphoric acid (3-dodecylmercapto-1-decyloxy) -2-propyl ester was converted from the corresponding alcohol (WO 91/05558) by reaction with POCI ₃ and subsequent hydrolysis produced and used as a raw product in the above reaction.

Example 2

(3'-Deoxy-3 '-azidothymidine) -5'-phosphoric acid-1,3- bis (dodecylmercapto) -2-propyl ester was prepared analogously to Example 1 in 29% yield, oil, R _F = 0.24 (CH ₂ Cl ₂ / MeOH 8/2), Rf = 0.61 (CH ₂ Cl ₂ / MeOH / H2θ 6.5 / 2.5 / 0.4) on Merck 5715 TLC plates, silica gel 60.

Example 3

Efficacy and tolerability in the friend virus leukemia model

Female Balb / c mice, 6-8 weeks old (Iffa Credo), were given 0.2 ml of a virus-containing spleen supernatant per day i.p. inoculated. The animals were i.p. daily from day 0 (start: 1 h after virus inoculation) to day 13. treated with the substance to be examined in doses of 6.25 mg, 12.5 mg, 25 mg and 50 mg per kg.

Before the start of therapy and on day 13, the parameters body weight and small blood count (WBC, RBC, Hb, Hkt, Plt) and on day 14 after killing the animals, the individual spleen weights were determined as parameters for viremia. Table: Influence of the test substances on the FV leukemia in vivo: Average spleen weight on day + 14 after virus inoculation

Substance dose (mg / kg / day) ¹⁾ Spleen weight (g) ²⁾ n

* ⁾ Therapy daily ip day 0 (+ lh) - day + 13; Day 14;

²⁾ X ^" + SEM, n = number of animals / group

The substances according to the invention are investigated according to the same scheme as for AZT. The results obtained show that the substances examined have a dose-dependent effect on virus-related splenomegaly and can therefore be used in the therapy of retroviral infections. Example 4

Efficacy in HIV-infected cell culture

Routine trials in the MT2 system in microtiter plates with at least 4 concentrations were carried out largely automatically (Biomek from Beckman) (standard deviation <5%). In parallel, both the toxicity (cells + substance) and the antiviral effect (cells + substance + virus) were determined.

MT2 cells were pre-incubated with the substance to be examined and infected with HIV-1 (HTLV-III-B, MOI 0.03). The supernatant was removed, replaced with medium (including substance) and incubated for 7 days.

This was followed by an evaluation according to the cytopathic effect (syncytia), MTT test (vitality of the cells) and transfer of the supernatant for new infection.

Claims

1. phospholipid derivatives of nucleosides of the general formula I,

CH ₂ -YR ₂

in the

R is a straight-chain or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, which may be substituted one or more times by phenyl, halogen, C -Cg-alkoxy, C -Cg-alkylmercapto, Ci-Cg-alkoxycarbonyl, C - Cg-alkylsulfinyl or C -Cg-alkylsulfonyl groups can be substituted,

R _{2 is} a straight-chain or branched, saturated or unsaturated alkyl chain with 1-20 carbon atoms, which optionally one or more times by phenyl, halogen, C -Ce alkoxy, C -Cg alkyl ercapto-, C -Cg alkoxycarbonyl- or Ci-Cg-alkylsulfonyl groups can be substituted,

X represents a valence line, oxygen, sulfur, sulfinyl or sulfonyl,

Y has the same meaning as X, where the two groups X and Y can be the same or different, Z can be oxygen or sulfur,

A can represent a methylene group or an oxygen atom,

Nuc can be a residue derived from a nucleoside derivative, and

their tautomers and their physiologically tolerable salts of inorganic and organic acids or bases.

2. Compounds of formula I according to claim 1, characterized in that Nuc is a nucleosidic radical of formula II or IV

is where

B is hydrogen or a hydroxy group,

R4, ^{R5 are} each hydrogen or one of the radicals R4 and R5 is halogen, a hydroxyl, a cyano or an azido group and also mean

R ₃ and R4 can represent a further bond between C-2 'and C-3'.

B represents a basic group of formula III

(purple)

CHIb)

(III)

(IIIc) lllld)

in which

Rg can be hydrogen, an alkyl chain with 1-4 carbon atoms or halogen,

Rg 'can be a hydrogen atom or a benzyl or phenylthio radical,

R7 can be hydrogen, an alkyl chain with 1-4 carbon atoms or halogen,

Rs can be hydrogen, an alkyl chain with 1-4 carbon atoms, halogen, or a hydroxy or an amino group,

R _{9 can be} hydrogen or an amino group, and

R o is hydrogen, halogen, Ci-Cg-alkoxy, C -Cg-alkyl mercapto, or an amino group, which can be mono- or disubstituted by Cι ~ Cg-alkyl, C -Cg- Alkoxy, hydroxy-C -Cg-alkyl and / or C ₃ -Cg-cycloalkyl, aryl, hetaryl, aralkyl or hetarylalkyl groups, which in the aryl or hetaryl radical may also be substituted by one or more hydroxy, methoxy or or alkyl groups or halogen may be substituted, or allyl, which may optionally be substituted with mono- or dialkyl or alkoxy groups,

or Nuc, a cyclobutane, Oxetanozin- or a nucleoside of seco-derivatives derived radical of the type -CH -CH ₂ - 0 CH ₂ -B, -CH ₂ -0-CH ₂ -CH ₂ -B or -CH ₂ -CH (CH ₂ OH) -0-CH ₂ -B, where B has the meaning given above.

3. Compounds of formula I according to claim 1 or 2, characterized in that Ri is a straight-chain C -Ci4- alkyl group which can be substituted by a Ci-Cg-alkoxy or a C -Cg-alkyl mercapto group.

4. Compounds of formula I according to claim 1, 2 or 3, characterized in that R _{2 is} a straight-chain C ₉ -C 14 alkyl group which can be substituted by a Ci-Cg-alkoxy group or a Ci-Cg-alkyl ercapto group.

5. Compounds of formula I according to any one of claims 2-4, characterized in that R ₃ and R4 each represent a hydrogen atom or R ₃ and R4 together represent a bond.

6. Compounds of formula I according to any one of claims 2-5, characterized in that R5 is a hydrogen or fluorine atom or represents a cyano or azido group.

7. Compounds of formula I according to any one of claims 2-6, characterized in that Rg represents a hydrogen atom or a C -C4 alkyl group and Rg represents a hydrogen atom.

8. Compounds of formula I according to any one of claims 2-6, characterized in that Rg represents a hydrogen or halogen atom or an amino group.

9. Compounds of formula I according to any one of claims 2-6, characterized in that R o represents a hydrogen or halogen atom, a Ci-Cg alkoxy group or an amino group.

10. A process for the preparation of compounds of formula I according to claims 1-9, characterized in that

a) a compound of the general formula V,

R ₂ -Y-CH ₂

in which R 1, R ₂ , X, Y, Z and A have the meanings indicated, with a compound of the general formula VI,

Nuc - OH (VI) in which Nuc has the meaning given above, preferably means a compound of the formula Via.

in which R ₃ 'represents hydrogen or a hydroxy group protected by an oxygen protective group familiar to the person skilled in the art and R4' u. R5 ¹ each represents hydrogen, halogen, an azido, a cyano or one of the radicals R4 'and R5' is a hydroxyl group protected by an oxygen protective group familiar to the person skilled in the art, or R ₃ • and R4 'is a further bond and B has the meanings given,

in the presence of a condensing agent and a tert. Nitrogen base, e.g. As pyridine or lutidine, in an inert solvent such as. B. toluene, or directly in pyridine to the reaction and after hydrolysis, if appropriate, splits off the oxygen protecting groups in accordance with the methods customary in nucleoside chemistry, or

a compound of the general formula VII

R ² -Y-CH ₂ CH ₃ in which R 1, R ₂ , X, Y, Z and A have the meanings mentioned above, with a compound of the general formula VI or Via, in which R ₃ ', R', Rs' and B have the meanings given , in the presence of phospholipase D in an inert solvent, such as. As chloroform, brings in the presence of a suitable buffer to the reaction and, if appropriate, splits off the oxygen protecting group after the reaction, in accordance with the methods customary in nucleoside chemistry.

11. Medicament containing at least one compound of formula I according to any one of claims 1-9 in addition to pharmaceutical adjuvants or excipients.

12. Use of compounds of formula I according to any one of claims 1-9 for the manufacture of medicaments for the treatment of viral or retroviral infections or of diseases caused by these infections.

13. A process for the preparation of medicaments according to claim 11, characterized in that a compound of the formula I is processed with customary pharmaceutical auxiliaries or excipients to give pharmaceutical dosage forms.