ITMI940356A1 - PHARMACOLOGICAL ACTIVITIES OF PURINA, THEIR USE AND PHARMACEUTICAL COMPOSITIONS THAT CONTAIN THEM - Google Patents

Description

"DERIVATIVES OF PURIN WITH PHARMACOLOGICAL ACTIVITY, THEIR USE AND PHARMACEUTICAL COMPOSITIONS THAT CONTAIN THEM"

The present invention relates to the use of purine derivatives with calcium antagonist and antiviral activity.

In particular, the present invention refers to the use of compounds of general formula (I)

NH

where is it

R is a linear or branched C ^ c ^ alkyl group, or a -Y-A group, in which Y is a linear or branched C ^ -c ^ g alkylene chain, optionally linked to group A through a sulfur atom;

A is a halogen atom, a -SR ^ 0 -NR-JR2 group, where

R ^ and R2 are independently a hydrogen atom, a phenyl group, a linear or branched C ^ -C ^ alkyl group, a C ^ -C ^ alkylcycloalkyl or alkylpolicycloalkyl group, or taken together with the nitrogen atom whose sine bonded form a nitrogenous heterocyclic group possibly containing in the heterocyclic ring one or more heteroathems selected from N, 0, S, said heterocyclic group being optionally substituted in any position by one or more groups independently selected from C ^ _ C4 linear or branched alkyl, C1 ~ C4 linear or branched alkoxy, hydroxy, armine, mono- (C ^^ alkyl armine, di- (C1-C4) alkyl armine; their salts with pharmaceutically acceptable acids,

their forms enant iomere, diastereomere and relative mixtures.

The general formula (I) described above also includes new compounds which constitute another object of the present invention, which therefore refers to the compounds of general formula (I) with the exclusion of the following compounds:

9 - (2-bromoethyl) adenine j

9- (2-benzyl) adenine;

9- (2-N, N-diethylaminoethyl) adenine.

Examples of linear or branched alkyl groups are methyl, ethyl, propyl, isoprcpyl, butyl, terbutyl.

C ^ -C ^ Q chain examples linear or branched alkylene methylene, dimethylene, trimethylene, tethramethylene, pentamethylene,

*

hexamethylene, decylene, 1-methylethylene, preferably dimethylene, trimethylene, tetramethylene, pentamethylene.

Examples of group C ^ -C4 linear or branched alkoxy methoxy, ethoxy, propoxy, iso-propoxy, butoxy, tert-butoxy, preferably methoxy.

Halogen atoms are chlorine, bromine, iodine.

Examples of heterocyclic groups sine heterocycles with 6 atoms in the ring, for example pyridine, pyrimidine, pyrazine, 1,3,5-triazine, paperi dina, piperazine, morpholine, thiairorpholine, preferably pyridine, piperidine; 5-atom heterocycles in the ring e.g. pyrrole, thiazole, oxazole, imidazole, pyrrolidine, thiazolidine, oxazolidine, imidazolidine, ben zimidazole, preferably imidazole, benz imidazole.

Examples of pharmaceutically acceptable acids are halogenhydric acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, oxalic acid, propionic acid, fumaric acid.

The use of the following compounds is particularly preferred:

9- (2-bromoethyl) adenine, (compound I);

9- (2-benzyl) adenine, (compound II);

9- (2-N, N-diethylanminethyl) adenine, (carpus III);

9- (2nmorpholinoethyl) adenine, (compound IV);

9- (2- (1-benzimidazolyl) ethyl) adenine, (compound V);

9- (2- (1-piperidinyl) ethyl) adenine, (compound VI);

9- (2-N- (1- (3-adamant yl) et ylarm n) ethyl) adenine, (compound VII);

9- {2- (5-1, 2,4-triazolyl) thioethyl) adenine, (compound VIII);

9- (2- (5- (1-phenyl) -1, 2,3,4-tetrazolyl) thioethyl) adenine, (compound IX).

It is currently the opinion of many researchers that the most effective means of preventing viral diseases is still active vaccination. However, it should be noted that there are several viral diseases which for different reasons can hardly be prevented by vaccination alone. There are, for example, infections caused by herpes virus, influenza virus and hepatitis.

As is known, [Piovi T. - Med. Biol. 1980 v. 58 N. 1. - p. 5-7; Collier L.H. , Ed. By L.H. Collier, J. Oxford, Landcn, Acad. Press. 1980, p. 273-285; Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158], the herpes virion contains 24 structural proteins that are difficult to purify from contaminating proteins and therefore it is difficult to prepare a highly effective vaccine, moreover it has not yet been well established which proteins induce the production of antiobodies active protective.

On the other hand, influenza viruses are less complex than herpes viruses in immunological terms, but have a wide range of highly variable serotypes and breasts, which makes seroprophylaxis difficult [Zhdanov V.M. Moscow, 1979. p. 3-5; Chetverikova L.K., Kramskaya T.A., Romeiko-Gurko Yu.R., Moscow, 1982, p. 18-22; Shotlissek K., Moscow, 1979, p. 34-42]. With regard to infections caused by the hepatitis virus, up to now there are no specific means for their treatment and the only measure to control the disease is active immunization with vaccines, but their production is also linked to the many difficulties. of obtaining highly pure antigens and anti-bodies. That is why the development of new synthetic preparations for the chemoprophylaxis and chemotherapy of viral infections is very topical.

It should be noted that during the development of chemotherapy of viral infections, many organic compounds of different chemical classes have been synthesized which are capable of inhibiting viral replication. However, only a few compounds have demonstrated potential antiviral activity and most of them consist of derivatives of nucleic bases (mainly altered nucleosides) and adamantine. Eseirpi of such derivatives are: adeninarabinoside (vidarabine), cytosinarabinoside, 5-iodine-2-deoxyuridine, 5-trifluoromethylthimidine, aràbinosiltimin, retrovire (zidovudine), amantadine (sirmietrele, midantane), rimantadine, and others. [Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158; Indulen M.K., Dzeguze D.R., Kalnberga R. Yu. et al. Minsk, 1981, p. 9-12; Chizhov N.P. Yershov F.I., Indulen M.K. Riga, Znatne, 1988 p. 171]. Over the past 10 years, new types of abnormal nucleosides have been synthesized which have been studied for their antiviral activities and introduced into medical practice. These sine acyclovir, ganciclovir, 9- (2,3-dihydroxypropyladenine and others in which the carbohydrate fragment is replaced by an aliphatic chain similar to the part of the ribose molecule or of other character [Indulen M.K., Dzeguze D.R., Kalnberga R.Yu, et al. Minsk, 1981, pages 9-12; Elicn G.B., Furman P.A., Fyfe J.A. et al. Proc. Nati. Acad. Sci. USA - 1977, v. 74 - N. 12 pages 5716-5720; Schaeffer H.J., Beauchamp L., Miranda P. et al. Nature 1978, v. 272 - n. 5654 -page 583-585; Scheiner P., Gear A., Bucknor A.M. et al. Nucleosides and Nucleotides 1989 v. 8 N . 8 - pages 1441-1451; Fletcher C.V. Invest. Drug Inform. 1988 v. 23 - n. 5 pages 5-12]. Up to now the list of antiviral compounds has increased, · it includes new compounds, such as carbovir , phamciclovir, penciclovir, AICA-ribosides, arbidol, ribamidyl, adapremin, deitiforina [Vince R., Hua M., Brownell J. et al. -Nucleosides and Nucleotides - 1989 - v. 8 - n. 5.6 pag. 1127 -1128; Anderson B.D., Chiang Ch. Y. - J. Pharm. Sc i. 1990 - No. 9 p. 787-790; Chizhcv N.P. - Leningrad, 1990 pages 4-9; Hodge R.A.V. Sutton D., Boyd M.

et al. Antimicrob. Agents Chemother. - 1989 - v. 33 - No. 10, p. 1765-1773; Markin V.I., Evseev A.A., Bogatikov G.V. et al. Problems of virology, 1990 n. 2 pages 151-152; Markin V.A., Pashanin Yu. G., Markov V.I. et al. Problems of virology - 1990 - N. 3 - pag. 237-240]. Among the compounds mentioned above, acyclovir and retrovir are widely used against herpes and AIDS [Schaeffer H.J., Beauchanp L., Miranda P. et al. Nature - 1978 V. 272 N. 5654 - pag. 583-585; Collins P., Bauer D.J. - J. Antimicrobiol, Chemotherapy 1979 V. 5 pag. 431-436] as well as amantadine and rimantadine against influenza [Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158; Galegov G.A., Vladyko A.S., Linitskaya G.L. et al. Problems of medicai chemistzy - 1981, n. 2 p. 251-254]. As is known from the literature, [Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158; Furrnan P.A., Clair M.H. Fyfe J.A. et al. J. Virol. - 1979 - V. 32 N. 1 pag. 72-77], the action of acyclovir and its analogues is based on the inclusion of an abnormal fragment in place of native guanosine rxbofuranoside in the biosynthesis of viral nucleic acid, which stops further replication of the virus. In this process, an important role belongs to the lateral aliphatic chain of acyclovir, capable of being phosphorylated, which ensures the insertion of the molecular insert into the nucleic acid chain.

However, the nucleic base derivatives and heterocyclic compounds of other sine lineages are known for their antiviral activity, but lack the fragments capable of being phosphorylated. In this case, the antiviral properties of these compounds can be realized in other ways [Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158; Muller W.E.G. - New York, Raven Press - 1979 pag. 77-149].

Rimantadine and breast amantadine effective only in the initial stage of the disease. Many publications [Kalnynia V.A., Feldblum R.L., Indulen M.K., Riga, Znatne, 1984 p. 158; Chizhov N.P., Yershov F.I. Indulen M.K. Riga, Znatne, 1988 - page 171; Chizhov N.P. - Leningrad, 1990 pag 4-9] report studies of the antiviral mechanism of action. However, many aspects still remain obscure and no correlation between the structure and antiviral activity has yet been found. Since no other synthetic antiviral compounds more effective than aciclovir, rimantadine and amantadine are currently available, the search for new highly effective antiviral compounds is of current interest, not only in the nucleic base line and adamantane, but also among others. classes of heterocyclic compounds, particularly among those that have been studied to a lesser extent with respect to their antiviral activity.

It has now been found, and it is the object of the present invention, that the compounds of general formula (1) are endowed with antiviral activity.

Some compounds described in the general formula (I) are known in the literature, but none of these have been described as having antiviral activity.

9- (2-bromoethyl) adenine has been described as an intermediate for the synthesis of purine-structured Burrte salts (CA: 113: 211936z), of acyclic nucleotide analogues with antiviral activity (CA: 111: 23866m), of compounds with cardiovascular activity (CA: 96: 218190r), of polymers containing nucleic bases with biological activity (CA: 93: 137969 <η>; CA: 92: 215682J; CA: 92: 112263G; CA: 92: 17360; CA: 89 : 204134).

9- (2-N, N-diethylaminoethyl) adenine is described in a publication relating to triacanthin derivatives.

9- (2-benzyl) adenine is described in numerous patents, mostly relating to the agricultural sector (ZA 8600757; JP 7529346; JP 7418661) and as anti-invasion agents (US 3930005).

Compounds of formula (I) are prepared with methods known in the literature, as for example described in the references cited above.

Briefly, the compound of formula (II)

NH.

where Y is defined above and X is a halogen atom, preferably chlorine, bromine, iodine, more preferably bromine, is reacted with a compound of formula HA, where A is as defined above (possibly in the presence of acidity acceptors) according to methods known to the average expert.

Compounds according to the present invention were tested for their antiviral activity in experiments on chicken embryos subjected to influenza A / Leningrad / 134/72 virus and on mice subjected to influenza A / Bethesda / 10/63 influenza virus. 1 influenza A / Aichi / 2/68.

The results are reported in the following Table 1.

TABLE 1 - Antiviral activity of compounds in experiments on chicken embryos subjected to influenza virus

A / Leningrad / 134/72 on white mice subjected to influenza A / Bethesda / 10/63 influenza

A / Aichi / 2/68.

Compound Activity index Comparison of

on experimental activity models

chicken white mice

Depression index Amantadine index Rimantadine Virazole protection infectivity protection (%) (%) {%)

(%) (lg EID50) (%) i

the

VI 29.0 46.0 52.0 44.0

to c n H - «+ IX 53.0 1.0 41.0 65.0 73.0 62.0 SU C P cu n H-ero Amantadine 63.0 100.0 113.0 95.0 fl> rt n r + O H- 3 Rimantadine 70 , 0 1.8 56.0 89.0 100.0 85.0 U)

Ci C H- 1— 1 c ro Virazole 66.0 105.0 118.0 100.0 in es (D N Ό tu P ro «CD fD

0) (+ H * rtrt · c M tu l · * · I— 1 (0 [/] »h The same activity was proven as a function of the contaminated dose (influenza A / Leningrad / 134/72 virus). The breast results

shown in the following Table 2.

TABLE 2

Compounds Activity index

Depression of the protective index according to viral infectivity the contaminated dose (lgEID5 {)) lgEID ^ g)

6-60 0.6-6.0 Vili 0.0 0.0 24.0

IX 1.0 33.0 53.0

Furthermore, the activity of the compounds of the present invention was tested in the Herpes virus center under extremely drastic conditions, ie in the encephalitis induced by Herpes Simplex A virus in mice. The following Table 3 reports the results.

TABLE 3

Body sto Activity index

Deaths% Protection Index Coefficient% IV 60.0 1.3 25.0

II 70.0 1.14 12.0

V 70.0 1.14 12.0

VI 60.0 1.2 15.0 Placebo 100, 0

Quite surprisingly, the compounds according to the present invention have also been shown to possess calcium antagonist activity. This activity is represented by the results reported in. Therefore, the use of compounds of formula (I) as antiviral agents and calcium antagonists is a further object of the present invention.

Furthermore, a further object of the present invention is the use of compounds of formula (I) as active principles for the preparation of a medicament with antiviral activity.

A further object of the present invention is the use of compounds of formula (I) as active principles for the preparation of a medicament with calcium antagonist activity.

The present invention also comprises pharmaceutical compositions containing a compound of formula (I) in admixture with pharmaceutically acceptable carriers and excipients. The compositions according to the present invention are prepared with conventional methods and procedures known to those skilled in the art, as described for example in "Remington's Pharmaceutical Sciences Handbook", XVII ed. , Mack Pub. , N.Y. USA.

Examples of pharmaceutical compositions include injectable forms, such as sterile solutions or suspensions or restorable lyophilized powders, oral forms, such as tablets, capsules, tablets, dragees, powders, granulates, syrups and solutions; rectal and vaginal forms, such as suppositories, ovules, candles; topical forms, such as creams, ointments, lotions.

Dosages and posology will be established by the attending physician according to the severity of the pathology to be treated and the patient's condition (age, sex, weight). Dosages will generally be between 0.001 and 100 mg / kg

The following examples further illustrate the invention.

EXAMPLE 1

9- (2- (5-1, 2,4-Triazolyl) thioethyl) adenine, (compound VIII) 0.61 g (0.004 mol) of 9-hromoethyladenine, 0.61 g (0.006 mol) of 5-mercapto- 1, 2,4-triazole, 0.5 g of ^ CO ^, 0.02 g of KI and 10 ml of ethanol were mixed and steamed in a sealed vial for 8 hours. After which the vial is cooled, opened and the solvent is concentrated under vacuum. The residue is washed with water and recrystallized from ethanol. 0.73 g (70%) of the title compound (CQHIQNSS) are obtained with m.p. 196 ° C.

IR spectrum (cm-1): 3300, 2500 (NH2 and NH), 1610, 1550 (-CH ^ CH- bond in positions 5 and 6 of adenine).

NMR spectrum (DM90, £): 7.40 and 8.10 (s, H-2 and H-8 of adenine), 8.20 (s, H-4 of triazole), 3.7 and 4.5 ( t, j-3.5 Hz, -N-CH2-CH2-S-). Protons of NH and NH2 are not detected in the spectrum due to exchange reactions with dimethylsulfoxide in which the spectrum was made.

EXAMPLE 2

9- (2-N- (1- (3-Adamantyl) ethylanmin) ethyl) adenine (compound VII) 0.61 g (0.004 mol) of 9-bromoethyladenine, 1.34 g (0.012 mol) of rimantadine and 5 ml of ethanol are mixed and steam heated in a sealed vial for 8 hours, after which the vial is cooled, opened and the solvent is concentrated in vacuo. Then 1 ml of 5% KQH is added and the whole is concentrated again. The residue is extracted with benzene, then the organic phase is dried over KOH, after which the benzene layer and the base are transformed into sulphate following Table 4, where the compounds according to the present invention are

were tested for their calcium channel blocker activity according to the model

study of CaClj-induced arrhythmia in mice. The members

of the invention were compared with known calcium antagonists.

TABLE 4

COMPOUND DOSE, TIME OF INTRODUCTION- ^ 50 LD50 WIDTH (rng / Kg) NE BEFORE CaCl- (min) mg / kg mg / kg THERAPEUTIC

NO. OF MICE 90PRAVIS-

SUTI%

2 min 5 min 15 min

CaC12 330.0 0.0

Finoptina 0.1 0.0 0.0 0.0

1.0 0.0 0.0 0.0

10.0 50.0 30.0 0.0 10.0 41.0 4.0 Diltiazem 0.01 40.0 40.0 20.0

0.01 60.0 60.0 40.0 0.1 181.0 1810.0 1.0 70.0 80.0 60.0

Nifedipine 0.01 0.0 0.0 0.0

0.1 20.0 20.0 0.0

1.0 10.0 20.0 0.0 1.0 190.0 190.0 Vili 0.01 40.0 20.0 30.0

0.1 90.0 60.0 30.0 0.1 890.0 8900.0 1.0 70.0 50.0 30.0

VII 0.01 100.0 50.0 30.0 0.01 100.0 10000.0 0.1 70.0 50.0 50.0

1.0 80.0 60.0 20.0

III 0.01 40.0 45.0 20.0

0.1 50.0 50.0 20.0 0.1

1.0 30.0 20.0 40.0

From the above, the compounds of the present invention are

endowed with pharmacological activity, in particular antiviral and calcium

antagonist.

by solution of sulfuric acid in ethanol. The residue is separated, washed with ethanol and dried. 1.72 g (98%) of the title compound (3⁄43⁄43⁄4) are obtained ∞n m.p. 231 ° C.

IR spectrum (cm-1): 3380, 2470 (NH2 and NH), 1610 (bond -CH = CH- in adenine).

NMR spectrum (CDC1, £ "): 7.40 and 7.90 (s, H-2 and H-8 of adenine), 6.30 (s, NH and ^ 2 ^ '^, 90 and 3.80 ( m, -N-Ci ^ -Ci ^ -N-), 1.50-2.0 (m, protons of the adamantane nucleus), 1.10 (d, j-3.0 Hz -CH ^).

EXAMPLE 3

9- (2- (5- (1-Phenyl) -1, 2,3,4-tetrazolyl) thioethyl) adenine (compound IX) 0.61 g (0.004 mol) of 9-brcmoethyadenine, 1.07 g (0.006 moles) of 1-phenyl-5-mercapto-1, 2,3,4-tetrazole, 0.5 g of I ^ COj, 0.02 g of KI and 10 ml of ethanol are mixed and steam heated in a sealed vial for 8 hours. The vial is then cooled, opened and the solvent is concentrated in vacuo. The residue is washed with water and recrystallized from ethanol. 1.0 g (74%) of the title compound (C14H13N9S) are obtained with m.p. 213eC.

IR spectrum (cm-1): 3340, 2510 (NH2), 1600 (bond -CH * = CH- in acfenine), 700 (C-S).

NMR spectrum (DM90, ÌT): 7.20 and 8.10 (s, H-2 and H-8 of adenine), 7.65 (m, aromatic protons), 3.85 and 4.60 (t, j -3.5 Hz, -N-CH2-CH2-S-).

Similarly, the following compounds were prepared

9- (2-morph olinoethyl) adenine (compound IV);

9- (2- {1-benzimidazolyl) ethyl) adenine (oonposite V);

9- (2- (1-piperidinyl) ethyl) adenine (compound VI).

Claims (7)

CLAIMS 1. Compounds of general formula (I): where is it: R is a linear or branched C ^ _c ^ alkyl group; yet a -Y-A group, in which Y is a linear or branched C - ^ - C- ^ Q alkylene chain, possibly linked to group A through a sulfur atom; A is a halogen atom, a -SR ^ 0 -NR-jI ^ group, where R ^ and R2 are independently a hydrogen atom, a phenyl group, a linear or branched alkyl group, a C3-C12 alkylcycloalkyl or alkylpolicycloalkyl group, or taken together with the nitrogen atom to which they are bonded they form a heterocyclic nitrogen group possibly containing in the 'heterocyclic ring one or more heteroatoms selected from N, 0, S, said heterocyclic group possibly being substituted in any position by one or more groups independently selected from linear or branched C ^ _ C4 alkyl, linear or branched C ^ -C ^ alkoxy , hydroxy, armine, mono-) alkylairmin, alkylithmino; their salts with pharmaceutically acceptable acids, their enantiomer, diastereoiscmere and related mixtures forms; with the exclusion of the following compounds: 9- (2-bromoethyl) adenine; 9- (2-benzyl) adenine; 9- (2-N, N-diethylaminoethyl) adenine. 2. Compounds according to claim 1 which are: 9- (2-morpholinoethyl) adenine; 9- (2- (1-benzimidazoles) letyl) adenine; 9- (2- (1-piperidinyl) ethyl) adenine; 9- (2-N- (1- (3-adamantyl) ethylamine) ethyl) adenine; 9- (2- (5-1,2,4-triazolyl) thioethyl) adenine; 9- (2- (5- {1-phenyl) -1,2,3,4-tetrazolyl) thioethyl) adenine. 3. Use in dogs with antiviral agents of the compounds of formula (I), NH 2 R. where is it: R is a linear or branched C ^ -c4 alkyl group; yet a -Y-A group, in which Y is a linear or branched C - ^ - C ^ Q alkylene chain, possibly linked to group A through a sulfur atom; A is a halogen athem, a -SR ^ ° -Ν3⁄43⁄4 '<^ group where RjL and R2 are independently a hydrogen atom, a phenyl group, a linear or branched C ^ -C ^ alkyl group, a C ^ -C ^ alkylcycloalkyl or alkylpolicycloalkyl group, or taken together with the nitrogen atom whose sine bond they form a nitrogenous heterocyclic group possibly containing in the heterocyclic ring one or more heteroatoms selected from N, 0, S, said heterocyclic group possibly being substituted in any position by one or more groups independently selected from linear or branched C ^ -alkyl, C ^ - C ^ linear or branched alkoxy, hydroxy, airmin, mono- (C ^ -c ^) alkylimino, di- (Cj-C4) alkylamino; their salts with pharmaceutically acceptable acids, their forms enanticmere, diastereoiscmere and related mixtures. 4. Use as agents with calcium antagonist activity of the compounds of formula (I), where is it: R is a linear or branched C ^ _c ^ alkyl group; or a group -Y-A, in which Y is a linear or branched C ^ -C ^ Q alkylene chain, optionally linked to group A through a sulfur atmosphere; A is a halogen atom, a -SR ^ 0 -NR-JR2 group, ctove 3⁄4 and R2 are independently a hydrogen atom, a phenyl group, a linear or branched alkyl group, a C3 ^ C12 alkylcycloalkyl or alkylpolicycloalkyl group, or taken together with the nitrogen atom to which bonded form a nitrogenous heterocyclic group possibly containing in the heterocyclic ring one or more heteroathems selected from N, O, S, said heterocyclic group possibly being substituted in any position by one or more groups independently selected from C ^ C4 linear or branched alkyl, C2-C4 linear or branched alkoxy, hydroxy , armino, mono- (C ^ ^ alkylanimino, di- (C1-C4) alkylamino; their salts with pharmaceutically acceptable acids, their enantiomer, diastereoiscmere and related mixtures. 5. Use of the compounds according to claims 1-3 for the manufacture of a medicament with antiviral activity. 6. Use of the compounds according to claims 1-2 and 4 for the manufacture of a medicament with calcium antagonist activity. 7. Pharmaceutical compounds containing one of the compounds of the preceding claims as an active principle.