CZ287795A3 - The use of a peptide derivative, combination of derivatives and pharmaceutical preparation - Google Patents

Abstract

Disclosed herein is a method for treating acyclovir-resistant herpes infections in a mammal. The method comprises administering a peptide derivative or a combination of the peptide derivative and an antiviral nucleoside analog to the infected mammal. The peptide derivative used for the method is represented by the formula A-B-D-CH2CH{CH2C(O)R<1>}C(O)-NHCH{CR<2>(R<3>)COOH}C(O)-E wherein A is a terminal group, for example an optionally substituted phenylalkanoyl, and B is an N-methyl amino acid residue; or A and B together form a saturated alkylaminocarbonyl; D is an amino acid residue; R<1> is, for example, an alkyl, cycloalkyl, or a monosubstituted or a disubstituted amino; R<2> is, for example, hydrogen or alkyl and R<3> is alkyl, or R<2> is hydrogen and R<3> is phenylalkyl, or R<2> and R<3> are joined to form a cycloalkyl; and E is a terminal unit, for example, an alkylamino or a monovalent amino acid radical such as NHCH(alkyl)C(O)OH.

Description

Use of the Peptide Derivative and the Combination of Derivatives of the αS -. / Pi pharmaceuticals - /

Df tn

Technical field

The invention relates to the use of a peptide derivative having an antiviral effect and to the combination of this derivative with other antiviral agents as well as a pharmaceutical composition containing them.

BACKGROUND OF THE INVENTION

Acyclovir is the most widely used active substance used for the treatment of herpes virus infections. Recently, however, there have been more and more reports of herpesviruses that are resistant to the agent, for example in P. A. Chatis et al., N. Engl. J. Med., 320, 297 (1989) and S. Safrin, Res. Virol., 143, 125, 1992. Recently, this type of resistant virus has been increasingly found in patients with severe HIV infection. Because of their reduced resistance, these patients become very sandno infected with the herpes sipmlex virus,

HSV type 1 and especially type 2.

Much attention was paid to the biochemical properties that characterize acyclovir resistant HSV isolates. The findings are presented in a summary publication on acyclovir resistant HSV by C. S. Crumpacker, J. Am. Acad. Dermatol., 18, 190, 1988. Resistant strains have functional changes in one or both of the enzymes encoded by the virus, these enzymes being thymidine kinase TK and DNA polymerase POL.

When exposed to acyclovir, cells infected with wild-type HSV catalyze the TK produced by acyclovir monophosphorylation on a much larger scale than cellular enzymes. The resulting monophosphate is then converted by cellular enzymes to triphosphorylated acyclovir. This material is then reacted further with the addition of a polymerase or a DNA polymerase. This results in a reduction of DNA synthesis viruKterá the tax collection-ávaiíieuíraui Tuji ^5-2 ..- 't .77' · '-ZV''' / * this způso'oem 7a-then-k.tomu occurs. that this DNA is happening

Two mechanisms of resistance have been described using viral thymidine kinase: 1) selection of thymidine kinase-free mutants, so that the activity of the enzyme after infection is very low, and 2) selection of thymidine kinase-producing mutants with altered substrate specificity in which the enzyme is able to phosphorylate thymidine but not acyclovir.

A third mechanism by which resistance can be generated and which involves DNA polymerase is the result of selection of mutants that encode an altered enzyme resistant to acyclovir triphosphate inactivation.

^_ - ---- Na pupil TADEO-resistance mechanism of HSV isolates can be resistant protřacycíovi ether classified as thymidindeficientní (TK) strains, strains with altered enzyme (TK) or strains with modified DNA polymerase (POL).

Despite having some ´highlyHi5stacks-if-healing_in ^ _ ”results. Due to the fact that lower anti-acylcir virus is used when the antiviral agent Foscernet is used, infection with a resistant virus type is increasingly recorded. This is a serious problem especially in AIDS patients as described in K. S.

Erlich et al., N. Engl. J. Med., 1989, 320, 293 and S. Safrin,

J. Acquired Immun. Defic. Synd., 5, suppl. 1, p. 29, 1992.

Thus, there is a need to find a means to treat these conditions.

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SUMMARY OF THE INVENTION It is an object of the present invention to provide a relatively safe composition for the treatment of herpes infections caused by types of virus resistant to acyclovir.

The peptide derivatives described by P.L. Beaulieu, R. Deziel, w. Moss and R. Planner in co-pending International Patent Application No. PCT / CA / 93 / OO95, filed March 12, 1993. The application relates to the use of various substances for the treatment of herpes infections. However, it is known that antiviral agents effective against herpes infections may not be effective against acyclovir resistant herpes simplex virus, as described, for example, in J.J. ÓBrien and D. M. Campoli-Richards, Drugs 37, 233, 1989, pp. 252-253. The finding that said peptide derivatives are indeed effective against acyclovir-resistant herpes simplex virus infections was surprising.

SUMMARY OF THE INVENTION

The present invention provides peptide derivatives of the general formula 1 abd-ch ₂ ch / ch ₂ c (o) r ¹ / c (o) -nhch / cr ² (r ³ ) cooh / c (o) -e (1) wherein

N-represents phenylacetyl, phenylpropionyl '(4-aminophenyl) propionyl, (4-fluorophenyl) propionyl, (4-hydroxyphenyl) propionyl, (4-methoxyphenyl) propionyl, 2- (phenylmethyl) -3-phenylpropionyl, 2 - / ( 4-fluorophenyl) methyl-3- (4-fluorophenyl) propionyl, 2 - [(4-methoxyphenyl) methyl] -3- (4-methoxyphenyl) propionyl, or benzylaminocarbonyl,

Β denotes (N-Me) -Val or (N-Me) -Ile or forms together

A and B a saturated alkylaminocarbonyl group, namely butylaminocarbonyl, 1-methylethylaminocarbonyl, 1-methylpropylaminocarbonyl, 1-ethylpropylaminocarbonyl, 1 _; lΛ matřivlbutvlam i nokarbony11-e thylbu tylam i ngkarb ony

1-propylbutylaminocarbonyl, 1-ethylpentylaminocarbonyl; ii) t-i-n-phenylenediyl-carbonyl; 1-ethylpentylaminocarbonyl;

2-ethylpentylaminocarbonyl, 1-methyl-1-propylbutylaminocarbnyl, 1-ethyl-1-propylbutylaminocarbonyl, 1,1-dipropylbutylaminocarbonyl (1-propylcyclopentyl / amino-4-carbonyl or (1-propylcyclohexylcarbonyl) carbonyl

D is Val, Ile or Tbg,

· R ¹ is 1-methylethyl, 1,1-dimethylethyl, 1-methylpropyl,

1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-methylcyclopentyl, NR ⁴ R ⁴ , where R ⁴ is hydrogen or lower alkyl and R is lower alkyl or form R and R together with the atom nitrogen to which a pyrrolidine, piperidine, morpholine or 4-methylpiperazine ring is attached, *

R is hydrogen and

--q-L ________________

R is methyl, ethyl, 4-methylethyl-11,1-l-dimethylethyl, propyl, 2-propenyl or benzyl, the carbon atom to which the R groups are bonded, and R has the R configuration; or ___>!

R ^{@ 2} and R ^@ 3 independently represent methyl or methyl or form a carbon atom. They are bound, cyclobutyl, cyclopentyl or cyclohexyl and? A

E is NHR, wherein R is 2-methylpropyl,

2,2-dimethylpropyl, 1 (R), 2,2-trimethylpropyl, 1,1,2,2-tetra-methylpropyl, 1 (R) -ethyl-2,2-dimethylpropyl, 2- (R, S) -methylbutyl, 2,2-dimethylbutyl, 3,3-dimethylbutyl, 1 (R), 2,2-trimethylbutyl, 1 (R), 3,3-trimethylbutyl, 2-ethylbutyl, 2,2-diethylbutyl, 2-ethyl- 1 (R) -methylbutyl, 2 = ethyl-2-methylbutyl, 1 (R) -ethyl-3,3-

dimethylbutyl, 2,2-dimethylpentyl, cis- or trans-2-methylcyclohexyl, 2,2-dimethylcyclohexyl or 7 'c' · '· ^1' · '! 11 (0¼Viyi changed E skunlnnu NHCHÍR) -7.

wherein the carbon atom to which R is attached has the configuration S, R is 1,1-dimethylethyl, 1-methylpropyl, 2-methylpropyl, 2,2-dimethylpropyl or cyclohexylmethyl and Z is CH, OH, C (O OH, C (O) NHgg or C (O) OR, wherein R is methyl, ethyl or propyl, as well as pharmaceutically acceptable salts of these compounds for use in the treatment of acyclovir-resistant herpes simplex infections.

; In a preferred embodiment of the invention, the peptide derivatives of formula (I) are those in which:

A is phenylpropionyl, 2- (phenylmethyl) -3-phenylpropionyl or benzylaminocarbonyl,

B stands for (N-Me) Val,

D stands for Tbg,

R 1 represents 1-methylethyl, 1,1-dimethylethyl, 1-methylpropyl,

... 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclopentyl, cyclohexyl or 1-methylcyclopentyl,

R is hydrogen and

R is methyl, ethyl, 1-methylethyl, propyl or benzyl, the carbon atom to which the groups are attached

3

R and R have an R configuration, or

R ² a

R ² independently represent methyl or ethyl or together with the carbon atom to which they are attached form a cyclobutyl, cyclopentyl or cyclohexyl group and represents a group NHR ~, wherein R is _{2,2-dimethyipřo:} pyl.LÍRlj 2 _To 2- ^ rimethyl. propyl is 1 (R) -ethyl-2,2-dimethylpropyl, 2,2-dimethylbutyl or 1 (R) -ethyl-3, .alpha.-dimethyl-butyl or is E (NHCHCR ') - Z''-' - The atom that is bonded to the α-group. ŘKmá kqnf iguraq IT namenárz72 ⁷ ^ - ip Olurertfty ropylr-ar-2k general represents CH ^. ^ O-Cl-FCU ΑΧΩΗλ_ C (O) _NH2 or C (O) OR ^f ethyl or propyl,

Wherein R is methyl, as well as pharmaceutically acceptable salts of these compounds, for use in the treatment of acyclovir resistant herpes simplex infections.

In another preferred embodiment, they are peptide derivatives of Formula 1 wherein:

A and B together form saturated alkylaminocarbonyl from the group 1-ethylpropylaminocarbonyl, 1-ethylbutylaminocarbonyl, 1-propylbutylaminocarbonyl, 2-ethylpentylaminocarbonyl, 1 = methyl-1-propylbutylaminocarbonyl, 1-ethyl-1-propyl butylaminocarbonyl, 1-propylbutylaminocarbonyl) aminocarbonyl a

D _T R \ R ^2, R ² and E have the meanings given vpředchozím paragraph, as well as pharmaceutically, other "with ol f *" tray h to "ingredient teach e n FIN-beta for use in treating herpes simplex infectious resistant against acyclovir.

A combination of a peptide derivative of Formula 1 or a pharmaceutically acceptable salt thereof may also be used

A pharmaceutically acceptable nucleoside analogue having antiviral activity or a salt thereof.

In combination, a nucleoside analogue having an antiviral effect can be used which can be enzymatically converted (in vivo) and / or inflammatory and / or to an alternative substrate for virus DNA polymerase. The nucleoside analogue having antiviral activity may be selected from known nucleoside analogues. Preferred nucleoside analogs for combination with a peptide derivative of the invention are, for example, acyclovir and analogs thereof, for example compounds of formula 2

CH ₂ OCH ₂ CH ₂ OH wherein R is hydrogen, hydroxy or amino, as well as salts thereof, acceptable from a pharmaceutical aspect. The compound of formula 2 wherein R is hydroxy is acyclovir.

Other preferred nucleoside analogues with antiviral activity useful herein include, for example, vidarabine, idoxuridine, trifluridine, ganciclovir, edoxudine, brovavir, fiacitabine, penciclovir, famciclovir and rociclovir.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures 1 and 2 are graphically depicting the results obtained by applying the isobol method to demonstrate synergy in the assay, e.g. - a peptide derivative of the general formula a 1 against herpes sim virus pi pi <= x, - resistant to β-cyclovirus. ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~.

The compounds of formula 1 can also be represented by formula 1

The term "amino acid residue" or "amino acid derivative" means a residue derived from an amino acid or amino acid derivative. alpha-amino acids by tranlating the hydroxyl group of the carboxyl group and one hydrogen atom of the alpha-amino group.

In general, the abbreviations used to designate amino acids and protecting groups are in accordance with the IUPAC-iJB Biochemical Nomenclature Commission Recommendation as outlined in European Journal of Biochemistry 138, 9, 1984. For example, Val, Ile,

Asp and Leu are L-valine, L-isoleucine, L-aspartic acid and L-leucine residues.

Asymmetric carbon atoms in the basic linear axis, that is, in the backbone of the peptide derivative of formula 1, except for the terminal groups A and Z (in group E). however, including the carbon atom bearing R ^z when E is NHCH (R) -Z are of the S configuration. However, exceptions apply to the carbon bearing the side chain CHgClCOR1 when R 6 is lower alkyl or lower cycloalkyl as mentioned above. In the case of this exception, the carbon atom has the R configuration.

The asymmetric carbon atom in the side chain of an amino acid or amino acid derivative, in terminal group A and terminal group E when E is NHR may be S or R.

Me, Et, Pr and Bu are alkyl, methyl, ethyl, propyl and butyl.

Groups Meet ETP ₂ C ^ C therefore represent for example 1-ethyl-l-methylpropyl and 1-ethyl-l-propyl-butyl.

The Tbg group is the (S) -2-amino-3,3-dimethylbutanoic acid residue. The gammaMeLeu group refers to the (S) -2-amino-4,4-dimethylpentanoic acid residue. The gamma-MeLeucinol group means (S) -2-amino-4,4-dimethylpentanol from which one hydrogen atom has been removed from the alpha-amino group.

Some other symbols are used during the application: (N-Me) Val for (S) -3-methyl-2- (methylamino) butanoic acid residue, (N-Me) Ile for (S) -3-methyl acid residue -2- (methylamino) pentanoic acid, (N-Me ^ Tbg for ~ ~ ... ... · · 10 (s) -2- (methylamino) -3,3-dimethylbutanoic acid, Asp (cyBu) for acid residue (S) -alpha-amino-1-carboxyeyclobutaneacetic and Asp (cyPh) for (S) -alpha-amino-1-carboxylic acid residue

Lower alkyl and lacquer are used in the lumen of the application as a single moiety or in combination with other moieties means a straight chain hydrocarbon moiety of 1 to 2 b atoms. The branched-chain residue contains azomethyl-1-methyl, 1-methylpropyl, 2-methylpropyl and 1,1-dimethylethyl.

1- (lower alkyl) - (lower cycloalkyl) means in the course of the application a lower cycloalkyl radical having a lower alkyl substituent at the 1-position, for example 1-ethylcyclopropyl, 1-propylcyclopentyl and 1-propylcyclohexyl.

Lower cycloalkyl, used as a single moiety or in combination with another moiety, means a saturated cyclic hydrocarbon moiety containing from 3 to 6 carbon atoms and is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

A pharmaceutically acceptable carrier is an inert carrier for the active ingredient which does not adversely affect it.

n ~ s kyy toxic active ingredient usually produced · - - --- Physiologically acceptable ,, ₌ nosič_ said backsheet pxijatelné J ₌ <osmetické vehicle comprising one or more non-toxic excipients which do not react with the active ingredient-ani- not reduce its effectiveness.

Effective amount means a predetermined antiviral amount of an antiviral agent sufficient for activity against viruses in vivo.

By a binding agent is meant an agent which aids in the formation of a dehydrative bond of a free carboxyl group in an amino acid or peptide with a free amino group of another amino acid or peptide to form an amide bond between the two reactants. Similarly, such an agent may assist in the formation of an acid-alcohol bond to form the corresponding esters. Said reagents induce or facilitate dehydrative coupling by activation of the carboxyl group. Suitable reagents of this type and groups activated by these reagents are described in conventional peptide chemistry textbooks, for example, in E. Schroder and KL Lubke, The Peptides, Vol. 1, Academic Press, New York, NY, 1965, pp. 2 to 128, and KD Kopple, Peptides and Minic Acids, WA 3enjamin, Inc., New York, NY, 1966, pp. 33-51. diphenylphosphoryl azide, 1,1'-carbonyldiimidazole, dicyclohexylcarbodiimide, N-hydroxysuccinimide or 1-hydroxybenzothiazole in the presence of dicyclohecylcarbodiimide. A very suitable and useful binding agent is (benzotriazol-1-yloxy) tris (dimethylamino) phosphonium hexafluorophosphate, described in B. Castro et al., Tetrahedron Letters, 1219, 1975, and also in D. Hudson, J. Org. Chem., 53, 617 (1988). The agent is used as such or in the presence of 1-hydroxybenzotriazole. Other useful and suitable agent is commercially available 2- (lH-benzotriazole-l-yl) -N, N, ^{N, N>} -tetramethyluroniumtetrafluorboritan.

Nucleoside analogues having antiviral activity and their pharmaceutically acceptable salts for use in the invention are known. As noted above, the individual compounds of this group are characterized in a manner that can mediate antiviral activity against herpes viruses, i.e., their ability to cause in vivo inhibition of DNA DNA polymerase of the virus. Important compounds of this group are acyclovir and analogs thereof, which have been described in U.S. Patent No. 4,199,574 (HJ Schaeffer) of April 22, 1980 and also in HJ Schaeffer et al., Nature (London), 272, 573. 1978 and T "A. Krenitsk al, Proc .-- ^^ ^ -Acad Natl. Sci .. _^> ^ US _81__ 3209 ,. 984. A compound of formula (2) wherein R @ 1 is hydroxy ^{- is} acylovirus 7 with the name chemistry O- (2-hydroxyethoxy). The compound of formula (2) in which R 'is hydrogen is 6-deoxyacylitrite, a chemical name of 2-amino-9 - [(1'-hydroxy)]. TBT (oxyoxymethylenediamine) ad-n-thiophenoxycyclo- _fc -Y from:

R 5n žnámenff ^ £ ^ Rio Riipi ^ A7 "j ^t er ^t čhe'm ^even! CKY 2w6 ^ ~ ^ h-SWI <> «5! ₌ í'Á.C2 .hxdrox3 ^^ ethoxy) methy1 / purine.

'' 9

It will be appreciated that the compound of formula 2 wherein R is hydroxy may exist in its tautomeric form, i.e., as 2-amino-1,9-dihydro-9 - [(2-hydroxyethoxy) methyl] -5H-purine- 6-one, the compound may be a mixture of both tautomeric forms, the percentage of each tau = tomer in the mixture depending on physical conditions. Tautomeric forms also occur with other antiviral antiviral analogs which contain an enolizable carbonyl group.

Other nuclides are exerted by the pivotiviral effect which can be used for the above-mentioned purpose, for example, dara-9 ', 9-beta-D-arabinofuranosadiene monohydrate according to R.J. Whitley et al., N. Engl. J. Med., 1982, 307, 971, idoxudine. 2'-deoxy-5-ioduridine. by_-W. Prusoff, Biochim. Biophys.

Acta, 32, 295, 1959, trifluridine, 2 ' -deoxy-5- (trifluoromethyl) - 'uridine according to US pateηίΈνέΚο ρ'Γ3ϋ ^ "' ^δΤ ί '" 3 "201,"^38; No. 7 - (= O -, - H -? - delber (R) - ger) dated August 17, 1955, ganciclovir, 9 - [(1,3-dihydroxy-2-propoxy) methyl] guanine, according to U.S. Pat. No. 4,355,032 (JP Verheyden and JC Martin) of October 19, 1982, edoxudine, 5-ethyl-2'-deoxyuridine according to U.S. Pat.

553,192 (K. K. Gauri) of January 5, 1971, brovavir, (E) -5- (2-bromovinyl) -2'-deoxyuridine according to Y. Benoit et al.

Eur. J. Pediatrics, 143, 198 (1985), fiacitabine, 2'-fluorodeoxy-5-iodiuridine according to B. Leyland-Jones et al., J. Infect. Dis., 154, 430, 1986, penciclovir, 9./4-hydroxy-3- (hydroxymethyl) butyl / guanine according to S. E. Fowler et al., Br. J. Clin. Pharmacol., 28, 236P, 1989, famciclovir, 9- [4-acetoxy-3- (acetoxymethyl) butyl] adenine according to R. A. V. Hodge et al., Antimicrob. Agents Chemotherap., 33, 1765, 1989 and Rococlovir,

9 - [(1,3-diisopropoxy-2-propoxy) methyl] adenine according to E. Winklemann et al., Arzneim-Forsch., 38, 1545, 1988.

Production of peptide derivatives of general formula 1

Peptide derivatives of formula (1) may be prepared by methods which involve conventional methods of peptide synthesis, for example by classical coupling of amino acid residues and / or peptide fragments in solution. These procedures have been described, for example, in E. Schrdder et al., Supra. Lubke, T. Peptides, Analysis, Synthesis, Biology, Textbooks, and E-Gross et al., Eds., Academic Press, New York, N.Y., 1979-1987, Vol. 1-8 and J. M. Stewart and J. D. Young, Solid Phase Peptide Synthesis,

2nd Edition, Pierce Chem. Co., Rockford, IL, USA, 1984.

A common feature of the above processes for the production of peptide derivatives is the protection of reactive groups in the side chain of various amino acid residues or derivatives thereof, or optionally non-peptide fragments of the peptide derivative using suitable protecting groups to prevent chemical reaction at said site. It is also common to protect the .alpha.-amino group on the amino or peptide fragment, with the remainder of the molecule reacting at the carboxyl group and, upon completion of the reaction, to selectively remove the .alpha.-amino protecting group so that further reactions can take place.

Another common procedure is the initial protection of the C-terminal carboxyl group in an amino acid or peptide fragment, a carboxyl group that is later to be the C-terminal function of the peptide derivative. K.protection .se. užije ,. A protecting group that prevents the chemical reaction at this point already after removal or deprotection of the desired peptide derivative.

The key is to be read in the form of the formula and the formula Ϊ is ^_ m.ezip'roduktdbecnéhb ' formula ~ 3 ³ ......

wd-ch ₂ ch (ch ₂ c (o) r ¹ ) c (o) oh (3) where

W represents an alpha-amino protecting group, for example tert-butyloxycarbonyl (Boc), benzyloxycarbonyl (Z) or fluoren-9-ylmethoxycarbonyl (Fmoc), and

D and are as defined above.

This material can be prepared by Michael addition of an allyl ester of the formula CH ₂ WD-C (0) 0CH ₂ CH = CH ₂ in fumaroylový derivative of the formula (lower alkyl) -OC (O) CH = CHC (O) R ''^'for _ to form a Michael addition compound of formula

WD-CH (C (0) _of 0CH = _CH) CH (CH, C (0) R ¹⁾ C (0) O-lower alkyl, d.

...... If it is, it is treated with tetrakistriphenylphosphine palladium and trienylphosphine in the presence of pyrrolidine according to the method of R-Deziel, Tetrahedron Letters, 28,

4371, 1987, hydrolysis followed by decarboxylation of the allyl ester to give the corresponding alkyl ester of the key intermediate. Hydrolysis of this ester in the presence of a base such as sodium hydroxide or lithium hydroxide provides the key intermediate as a mixture of diastereoisomers.

A key intermediate of formula 3 wherein W and D are as defined in the preceding paragraph and R ¹ is a R 5 R 5 group wherein R and R are lower alkyl radicals or form together with the nitrogen atom to which they are attached a pyrrolidine , the piperidine, morpholine or 4-methylpiperazine moiety may also be prepared by reacting the above allyl ester of the formula WD-CH ₂ C (O) OCH ₂ -CH = CH ₂ , wherein W and D are as defined above. reaction under Michael reaction conditions with a fumaroyl derivative of formula BzlOC (O) CH = CHC (O) OBzl, i.e. with (E) -2-butenedicarboxylic acid dibenzyl ester to give the corresponding Michael addition product of formula WD- (RS) -CH ( CH (C (O) OBz1) CH ₂ C (O) OBzZ) C (O) -OCH ₂ CH = CH ₂ .

When treated with tetrakistriphenylphosphine palladium and triphenylphosphine in the presence of the pyrrolidine according to Deziel, the corresponding dibenzyl ester of formula WD-CH ₂ - (R, S) -CH (CH ₂ C (O) OBzl) is obtained. C (O) OBzl. Subsequent hydrogenolysis in the presence of palladium hydroxide on charcoal gives the corresponding dicarboxylic acid, which is converted to the corresponding anhydride by heating with acetic anhydride in excess. This anhydride is reacted with the appropriate secondary amine in the presence of pyridine to give a mixture of isomers predominantly of the desired isomer of formula WD-CH ₂ - (RS) -CH (CH ₂ C (O) NR ⁴ R ⁵ ) -C (O ) -0H, wherein each symbol is as defined above. This material was then converted to the benzyl ester and some was separated by HPLC to give the desired product as the desired isomer. Hydrogenation of this product in the presence of palladium hydroxide on charcoal gives a key intermediate of formula 3 wherein W and D are as defined above and R is NRR as defined above.

In general, therefore, a peptide of formula (I) may be prepared by sequentially sequencing the amino acid residues or derivatives thereof or non-peptide fragments of the peptides, for example, key intermediates, in sequence, depending on the desired peptides. The closure of the vessel is increased. All of the protecting groups are then removed from the formula (1). In the following example, the compounds of formula (I) may be prepared in the form of a pharmaceutically acceptable salt. . Where a p-peptide contains a basic group, examples of such salts include salts with organic acids such as acetic, lactic, succinic, methanesulfonic or p-toluenesulfonic acid, or also with polymeric acids such as tannic acid or carboxymethylcellulose, in addition, they may be salts with inorganic acids such as hydrohalic acids, for example hydrochloric acid or sulfuric or phosphoric acid. If desired, the acid addition salt of a particular acid may be converted into an acid addition salt of another acid, for example, a nontoxic, pharmaceutically acceptable salt by treatment with an ion exchange resin as described by R. A. Boissonnas et al., Helv. Chim. Acta, 43, 1849, 1560.

In cases where the peptide content is determined. One or more free carboxyl groups may be exemplified by salts on the carboxyl group of salts with sodium, potassium or calcium cations, or salts with organic bases, for example triethylamine or N-methylbutyric acid. = - 17

Effectiveness against herpetic belief

The antiviral activity of the peptide derivatives of formula 1 can be demonstrated by biochemical, microbiological and biological assays for inhibition of the replication of acyclovir-resistant herpes simpex viruses type 1 and 2 (HSV-1, HSV-2).

The peptide derivatives of formula (1) using cell cultures have been described by T. Spector et al. v Why. Nati. Acad. SCi. USA, 82, 4254 (1985).

The therapeutic effect of the peptide derivatives can be exemplified by the test on inoculated animals (e.g. mice) with herpes simplex virus, causing eye herpes disease and used in testing antiviral agents. This assay has been described by C.R.Brandt et al. in J. Virol. Meth., 36, 209 (1992).

The peptide derivatives of the invention or salts thereof with therapeutic effects are administered locally or totally to warm-blooded animals (pigs, horses or humans) bound to a vehicle containing an amount of one or more carriers corresponding to the solubility and chemical properties of the peptide derivative, route of administration and current practice. . 0.1-5% and 0.5-2%, respectively, of the active ingredient are bound to the pharmaceutically acceptable carrier when administered topically in a dosage form. Suitable topical dosage forms are solutions, ointments and lotions.

For total administration, the peptide derivatives are administered either orally or intravenously, subcutaneously, intramuscularly bound to pharmaceutically acceptable carriers or carriers. For injection, a peptide solution in sterile water containing other solutes such as buffers,

18 preservatives and the corresponding amount of β-pharmaceutical-viral-viral-moieties or glucose-isotonic solution.

Suitable vehicles or carriers in the aforementioned dosage forms are described in Remington with a pharmaceutical composition which is incorporated by reference; Ptiblishing ^^. Company / Easton? Perm., 1990.

^ ~ ceštŠ pddániú 'on-the selection of the active substance contained in a dosage form ^^^ ^ lo ^ a ^ to ^ ^ vyyolstěieAonemociiSrií type ₌ Pro ~~ jcinarpdfflalýšr-raising dávek- ^ ^ ^ to ^ until it reaches the desired effect. The peptide derivative is administered at a concentration that provides a sufficient antiviral effect and does not yet cause unwanted side effects.

When administered topically, the dosage forms are applied directly to the infected area of the body, i. skin, eye, mouth or genital landscape in an amount sufficient to cover the entire infected area. This procedure is repeated after 4-6 hours until the lesion disappears

For total administration, the daily dosage amount of the peptide derivative of Formula 1 is in the range of 1.0 mg to 10 mg per kilogram of body weight per day. The optimal range is 1.0 mg to 5 mg per kilogram of body weight per day.

For the treatment of herpes viral diseases, the above preparations are safe and sufficiently effective. Concomitant administration of other anti-viral preparations, such as antiviral nucleosides, surface-active agent additions, or antiviral interferons, as described by S.S.Asculai and F.Rapp in US Patent 4, 507, 281, March 26, 1985, is also possible.

In the treatment of acyclovir-resistant herpes viral diseases, it has been found that concomitant administration

synergistic enhancement of the active ingredient of the present patent without undesirable toxic side effects. Therefore, in the treatment of acyclovir-resistant herpes infections in mammals, a combination of analogs is coupled to a pharmaceutically and veterinarily acceptable carrier. of anti-viral neocyclic derivatives of the active salts with the peptide derivatives of the formula I or their therapeutically active salts.

The term synergistic effect is used for the relationship antiviral and antiherpetic efficiency aforementioned combination nucleoside analog and the peptide derivative and _<i of formula 1 means that the antivirus and antiher15 Petický effect of the combination is higher than the expected effect of the sum of the actions of the individual components of combination - application of the active compound combination in the treatment of acyclovir-resistant herpetic diseases to warm-blooded animals (horses, pigs or humans) using a vehicle composed of one or more carriers in an amount determined by the desired solubility, chemical properties of the nucleoside analogue and peptide derivative of formula 1 and a relative amount of both active ingredients guaranteeing their synergistic antiviral effect. A preferred route of administration is the topical administration of a combination of both active ingredients (i.e., an antiviral nucleoside analogue and a peptide derivative of Formula 1 or a therapeutically active salt thereof) in the form of solutions, emulsions, ointments or lotions in pharmaceutically acceptable carriers. Such dosage forms may contain 0.01-1.0% by weight of the nucleoside analogue or therapeutically active salt thereof, or about 0.05 to 1% by weight of its therapeutically active salt.

In any other case, the active ingredients are in combination in an amount that guarantees a synergistic melting rate.

The following examples further describe the present invention. Temperatures are indicated in the Celsius scale. --n-- Q2toků _r ^ J ^ At the same uvedáná objeínová'-percent · - or. _their.

unless otherwise specified. Spectrum; The nuclear magnetic resonance was detected on a Bruker spectrometer at 200 MHz or 400 MHz (data was obtained at 400 MHz). The chemical shift (δ) is reported in ppm. Abbreviations used: BOC - tert-butyloxycarbonyl; Bu-butyl; Bzl - benzyl; DMF - dimethylformamide;

Et - ethyl; EtOH-ethanol; EtOAc-ethyl acetate;

Et ₂ O-diethyl ether; Me-methyl; MeOH-methanol;

Pr-propyl; TLC - thin layer chromatography;

THF - tetrahydrofuran.

* Example 1

General Procedure for Binding Reaction * (see also R. Korrorr et al., Tetrahedron Letter, 30, 1927 (1989)).

The first reactant, i.e. a free amine (or its hydrochloride) is dissolved in methylene chloride or an ACE TNI tril ~ and V it kl y 'extended' ťoJc ~ the "Tčft tunes"π; and ^ tep 1 ol * "4 ⁹ C To a stirred of solution under nitrogen was added 4 equiv. N-methylmorpholine. After 20 minutes, 1 eq. a second reagent, i.e. free carboxylic acid, and 1.05 eq. of a binding agent. (Suitable and effective binders in this case are (benz21 triazol-1-yloxy) tris (dimethylamino) phosphonium hexafluorotetramethyluronium tetrafluoroborate. The reaction is monitored by thin layer chromatography. After completion of the reaction, the methylene chloride (or acetonitrile) is evaporated under reduced pressure. Dissolve in ethyl acetate and wash the solution sufficiently with 1N aqueous cicrondic acid solution, aqueous sodium unxicican solution and saturated sodium chloride solution.

The organic portion was dried over magnesium sulfate, filtered, and concentrated to dryness under reduced pressure.

The residue, after absorption on silica gel, is purified by flash chromatography according to the procedure of W. C. Still et al. In J. Org. Chem., 43, 2923 (1978).

in

Example 2

Preparation of intermediate Η-Asp (cyPn) (Bzl) -NH- (S) -CH (CH _g CMe ₂ ) CH 2 OBzl ²⁰ Part a): (S) -g-Azido-1 - ((phenylmethoxy) carbonyl) - cyclopentaneacetic acid The compound was prepared from 2-oxospiro [4.4] nonane-1,3-dione according to the procedure of MN Aboul-Enein et al. in Pharm. Acta Helv., 55, 50 (1980) and using the asymmetric azidation described by DAEvans et al. J. Amer. Chem. Soc., 112, 4011 (1990).

A specific procedure is to add dropwise 469 mL (750 mmol) of a 1.6 M solution of butyllithium in hexane to a solution of 96.8 g (750 mmol) of solution 4 (S) - (1-

3 ° -methylethyl) -2-oxazolidinone in tetrahydrofuran (described by L. Prgen and J. Prol in J Org. Chem., 54, 3231 (1989)) under an argon atmosphere at -40 ° C. The mixture was stirred at -40 ° C for 30 minutes and then cooled to -40 ° C

- ..................... I

- 22 - 1 temperature -78 ° C. - 3-Dione was added dropwise to the cooled mixture and the mixture was stirred at 0 ° C for 1 hour. To the mixture was added (600 mL), 20% by volume aqueous solution of citric acid, the organic layer was separated and · the aqueous extracted with t tote formate rt ^^ ^ Sloučenéřjorganické ppdily- _± washed with brine, dried over 'sodium hořecnaxým and concentrated under -se · - -------- form-3G'0-g ~ rů2ové pevné- substance-3-42- _(1-M karboxycyklp- _____ _ _3S l'g _Mn = _psntyl cxcet yiiXsil ⁷ -Ethyl-4'-oxazole idinone,

- - - - - - - - - - - - - - - - - - -

1 acetonitrile and after addition of 128.3 g (89.2 ml, 750 mmol) of benzyl bromide and 114 g (112 ml, 750 mmol)

1,8-diazobicyclo [5.4.0] -undec-7-ene was stirred under argon for 16 hours. The volatiles were removed under reduced pressure and the residue was dissolved in a mixture of water and ethyl acetate. The organic portion was separated, washed with 10% aqueous citric acid solution, saturated sodium chloride solution, dried over magnesium sulfate and concentrated to dryness under reduced pressure to give an oil. Crystallization of the oil from a mixture of hexane and ethyl acetate gave 204 g of a white solid corresponding to benzyl ester. A solution of 70 g (187 tetrahydrofuran was cooled in a cooled solution was 0.66 M solution (189 mmol) in a yield of 73%). in 200 mL at -78 ° C. Within 15 minutes, 286 ml _

1,1,3,3,3-hexamethyldisilazane in tetrahydrofuran containing a 6% cumene solution. The resulting mixture was stirred at temperature for 45 minutes

-78 ° C and a solution of g (216 mmol) of 2,4,6-triisopropylbenzenesulfonylazide in 100 ml of anhydrous tetrahydrofuran was added in one portion to the cooled solution, and after 2 minutes 50 ml (860 mmol) of glacial acetic acid was added. The mixture was warmed and stirred for 1 hour at 35-45 ° C. Share

The volatiles were removed from the reduced pressure and the resulting yellow residue was triturated with 1.7 L of 4: 1 hexane: ethanol to give a white solid which was collected by filtration. The filtrate is mixed with 230-240 mesh silica. The volatiles were removed under reduced pressure and the resulting solids dried under reduced pressure at 30 ° C to remove the cumene. After absorption onto a silica column, the solid was eluted with a 9: 1 mixture of hexane and ethyl acetate. Concentration of the eluate yielded 66 g of 3- ((2 (S) -azido-1-oxo-2- (1- ((phenylmethoxy) carbonyl) cyclopentyl) ethyl) -4 (S) - (1-methylethyl-2-oxazolidinone) yield 86%.

A solution of 13.42 mg (32.4 mmol) of this compound in 608 mL of 3: 1 tetrahydrofuran / water was cooled to 0 ° C. To the cooled solution was added 16.3 mL (518 mmol of hydrogen peroxide) of a 3: 7 mixture of hydrogen peroxide and then 2.86 g (68.2 mmol) of a mixture of lithium hydroxide and water. After stirring at 0 ° C for 45 minutes, the reaction is quenched by addition of 400 mL of 10% aqueous sodium sulfite. After addition of 1.93 g of sodium bicarbonate, the mixture was concentrated under reduced pressure. The chiral excipient is recovered for 20 hours by continuous extraction from a mixture of aqueous sodium bicarbonate and chloroform. After cooling the aqueous portion to 0 ° C, it is brought to acidic pH by addition of concentrated hydrochloric acid and then extracted with ethyl acetate. The extract was washed with a saturated sodium chloride solution, dried over magnesium sulfate and concentrated under reduced pressure to give a white solid.

8.2 g of the title compound as a white solid in a yield

84%. ¹ H NMR (CDCl 3): δ 1.6-1.8 (m, 5H), 1.95-2.05 (m, 2H), 2.20-2.30 (m, 1H), 4.55 (s, 1H), 5.12 (s, 2H) and

24.4 (m, SH). This compound was used in the synthesis of ^ sequent _{slGUČe-i-Nina} šůstl - GM hQto_přikladu ^ ₁ _

Part b): NH _; - (S) -CH (CH ₃ CMe) CH ₂ OBzl

The reduction of H-γMeLeu-OH is carried out using a mixture of lithium borohydride (LiBH ₄ ) and trimethylsilyl chloride (Me ₃ SiCl) according to the procedure of A.Giannis et al., AnhWin in Αηηβν. chí ^ m. Tnt ... Fd - η.ησί. 28, 218 (1989) to give the aminoalcohol of formula NH ₂ - (S) -CH (CH ₂ CMe) CH, OH. A mixture of 812 mg (6.2 mmol) of the above

- '-.

Of compound (I), 659 mg (/ (51 mmol)) of triethylamine and 1.42 g (6.51 mmol) of di-tert-butyl bicarbonate in 15 ml of tetrahydrofuran are stirred under nitrogen for 15 minutes at 4 °. After evaporating the tetrahydrofuran under reduced pressure, the resulting residue was dissolved in ethyl acetate and the solution was washed with 10% aqueous citric acid, 5% aqueous sodium bicarbonate and saturated sodium chloride. over magnesium sulphate and concentrated to dryness under reduced pressure, and the residue is purified by flash chromatography on silica, eluting with hexane:

2: 1 ethyl acetate to give 1.23 g of Boc-NH- (S) -CH (CH ₂ CMe ₃ ) CH ₂ OH in 86% yield.

To a solution of 1.23 g (5.35 mmol) of Boc-NH- (S) -CH (CH ₂ CMe ₃ ) CH ₃ OH in 13 mL of benzyl chloride was added successively 106 mg of tetrabutylammonium bisulfate and 3 mL of 50% aqueous solution. The hydroxide was stirred at 35-40 ° C for 90 minutes, diluted with ethyl acetate and washed with water and saturated sodium chloride solution. The organic portion was dried over magnesium sulfate and concentrated to dryness under reduced pressure. After the residue is dissolved in hexane, the resulting solution is poured onto a silica column and the column is eluted with hexane to

In this case, the benzyl chloride is removed. Elution with 2: 1 hexane: ethyl acetate gave Boc-NH- (S) -CH2-.

- (CH ₂ CMe ₃ ) CH ₂ OBzl. ¹ H NMR (CDCl ₃ ) of this compound: δ 0.95 (s, 9H), 1.42 (s, 9H), 1.30-1.55 (m, 2H), 3.42 (d, J = 4 Hz, 2H), 3.88 (br. 1H), 4.54 (m, 3H),

7.23 - 7.4 (Μ, 5H).

In 10 ml of a 6N solution of hydrochloric acid in dioxane, J2O ZptlS was added. χ μ \ u / J? The solution was stirred under nitrogen at 4 ° C for 45 minutes.

Removal of the solvent by evaporation gave the title compound (1.05 g), which was used in the next section of this Example without further purification.

Part c): Preparation of the resulting compound of this example ₁₅ H-Asp (cyPn) (Bzl) -NH- (S) -CH (CH ₂ CMe ₃ ) CH ₂ O Bzl

The compound is prepared by a coupling reaction wherein the starting compound is the resulting compound prepared in Example 1, the first reagent is NH ₂ - (S) -CH (CH ₂ CMe ₃ ) CH ₂ OBzl and the second reagent is the resulting "compound of part a) of this of the example, ie (S) -? - Azido-1 - ((phenylmethoxy) carbonyl) cyclopentaneacetic acid. N - ((S) -1-benzyloxymethyl-3,3-dimethylbutyl) is formed -

- (S) -? - azido-1 - ((phenylmethoxy) carbonyl) cyclopentane acetamide. Reduction of this compound in the presence of stannous chloride in methanol gives the following procedure by N.Maiti et al. in Tetrahedron Letters, 27, * -, 1423 (198-6), the resulting compound of this example. ^{Χ Η} NMR (CDCl) of this compound: δ 0.98 (s, 9H), 1.22 to 2.25 (s, 12H), 3.4 (d, J = 4 Hz, 2H), 3.64 ( s, 1H), 4.18 (br; m, 1H), 4.52 (m, 2H), 5.12 (s, 2H), 7.18 (d, J = 730)

Hz, 1H), 7.22-7.38 (br _; m, 10H).

AND

- 26 Example 3

-Preparation of Intermediate Boc-THG-FRS-Cří7 ^ ~) ~ CH7-GH ^ e (Q) -CMe ₃₎ C (O) OH A) magnesium salt monoalylmalonatu ..... Ro ztok. 10 0. g. (0.69 mol) of 2,2-dimethyl-1,3-dioxane-4,6-dione and 47 ml (0.69 mol) of allyl alcohol 0C, -xl

-> Λ 2ϊ_ΊΤί · ^; After removal of the solvent under reduced pressure, any residue is distilled under reduced pressure to give ^ 71% of mono-ammonium malonate in a yield of ?1% and a melting point of 123-127 ° C at 7.02 kPa. 71 g (0.48 mol) of this ester are dissolved in 300 ml of anhydrous tetrahydrofuran and 28.5 g (0.245 mol) of magnesium ethoxide are added. The mixture was stirred under argon at room temperature (20-22 ° C) for 4 hours, the solvent was removed by evaporation under reduced pressure, and the residue was triturated with ethyl ether to give a tan solid. After grinding the solid to fine particles and drying under reduced pressure, 56 g of the monoalyl malonate magnesium salt are obtained in a yield of 73%. This compound was used in the following part of the example.

Part b) Boc-Tbg-CHgC (O) OCH? CH _F

12.6 g (78 mmol) of 1,1'-carbonyldiimidazole are added to a solution of 15 g (64 mmol) of Boc-Tbg-OH in 150 ml of anhydrous acetonitrile. '

24 g (78 mmol) of monoalyl malonate magnesium salt and 100 mg of 4- (N, N-dimethylamino) pyridine were added to the mixture at room temperature under argon. The mixture was heated under reflux for 1 hour and stirred at room temperature for 18 hours. The mixture was concentrated under reduced pressure and the resulting residue was dissolved in 300 mL of ethyl acetate.

The solution was washed twice with 100 ml of 10% aqueous citric acid solution and twice with 100 ml of saturated solution

The sodium chloride is dried over sodium sulphate and concentrated to dryness by evaporation. The resulting residue was purified by flash chromatography on silica eluting with 3: 1 hexane / ethyl acetate to give the title compound as a brown oil (19.4 g), which crystallized on standing in 96% yield.

1 H NMR (CDCl ₃ ): The compound is a mixture of ceco-enolytic tautomers in chloroform in a ratio of 3 ° 1. Δ 0.96 (s,

9H, enol form), 1.04 (s, 9H), 1.46 (s, 9H), 3.65 (s,

2H), 3.90 (d, J = 8.5 Hz, 1H, enol form), 4.20 (d, J =

7.5 Hz, 1H), 4.65 (s, 2H), 5.10 (br * d, J = 7.5 Hz, 1H), 5.20-5.40 (m, 2H), 5.80- 6.05 (m, 1 H), 12 (s,

1H, enol form). This allyl ester was used to prepare the title compound in part d) of this example.

Part c): (E) -5,5-Dimethyl-4-oxo-2-hexenoic acid ethyl ester

The synthesis procedure is described by S. Manfredini et al. in Tetrahedron Letters, 29, 399-7 (1988). The crude product as an oil was purified after absorption on silica by flash chromatography eluting with hexane to give a yellow oil of the resulting ethyl ester, which was used in the next section of this example.

1 H NMR (CDCl 3) of this compound: δ 1.21 (s, 9H), 1.33 (t, J = -7.5 Hz, 3H), 4.28 (q, J = 7.5 Hz, 2H) 6.78 (d, J =

15.5 Hz, 1H), 7.51 (d, J = 15.5 Hz, 1H).

Part d): Boc-Tbg-CH; - (RS) -CH (CH; C (O) - CMe ₃ ) C (O) OH

the resulting compound of this example

0.67 g (2.1 mmol) was dissolved in 25 ml of anhydrous tetrahydrofuran under argon.

Boc-Tbg-CH.C (O) O-CH ₂ = CH ₂ (resulting compound of part b)).

To the solution was added 0.095 g (2.4 mmol) of a 60% dispersion of sodium hydride in oil. The mixture was stirred at rt for 30 min

(4-oxo-2-hexenoic acid ethyl ester) (described in Part c) of this (E) -5,5-dimethyl-4-oxo-2-hexenoic acid ester (described in part c) of this example). The reaction mixture is stirred until the reaction is complete (about δ hours), which is graphically thin-layered.

terminate by the addition of a 10% aqueous citric acid solution, ocθΰΙάΓΑγΌΙΟΙίχΙαίχ oc OQo ΰΙάηΖ 'Ζά ο'ΠχΖ'Θ'ίΤΘΤχΟ ΰΧα-Κΐϊ S ^- > λγ · 7η7v · Ί vs ° —vJc — álx - γ-xsJmxjs 25 s xsJmxjs 25 The state of the formula in H 2 O 4 is less than 5%. »X-i m λ. t-. -, x-4 «-» 4 · ** «« i

- Ur of *. ’Cš * IC * V i> -_ * ~ _Λ ~ - ~ V * ί. —A— · ~ ίΓ; - £ ΛθΛ-Α— -A. The residue is purified after absorption on silica, and the residue is purified after absorption into silica. flash chromatography eluting with hexane: ethyl acetate 9: 1 to give 1.06 g of the Michael addition product in 100% yield.

Conversion of the addition product of Michael 's reaction to

Boc-Tbg-CH- (RS) -CH (CH ₂ C (O) -C Me ₃ ) C (O) OH:

0.20 g (0.18 mmol) of tetrakistriphenylphosphine palladium (0) and 0.060 g (0.23 mmol) of triphenylphosphine are dissolved in 10 ml of methylene chloride under argon.

After addition of 20 ml of acetonitrile, the solution is cooled to 10 DEG C. and steam is added to OTSB3 (277 mmol of pyrrolidine and 1.06 g of Michael addition product) and the mixture is allowed to warm to room temperature over 1 hour. and then stirred for 20 hours under an argon atmosphere, and the mixture ^is refluxed for 1 hour.

The solvent was removed by evaporation and the resulting residue was purified by flash chromatography on silica eluting with hexane / ethyl acetate (9: 1) to give 0.77 g (1%, m.p. 7 mmol) Boc-Tbg-CH- (RS) -CH (CH ₂ C (O) CMe ₃ ) C (O) OEt in 83% yield. The entire amount of this compound was dissolved in 10 ml of a 1: 1 mixture of ethylene glycol dimethyl ether and water, and then 0.31 g (7.4 mmol) of sodium hydroxide-lithium monohydrate was added to the solution. The solution was then brought to acidic pH by addition of 20 ml of a 10% aqueous citric acid solution and extracted three times with 25 ml of ethyl acetate. The extract was dried over magnesium sulfate and concentrated under reduced pressure to give 0.69 g of a tan solid, yielding 80% (based on Boc-Tbg-CH ₂ C- (O) O-CH ₂ CH = CH ₂ o). according to NMR, is a mixture of diastereomers in a ratio of 55: 45. ^X 1 HNMR (CDCl ₃₎ of this compound: δ 0.97 (s, 9H, second isomer) ,. 0.99 (s, 9H, major isomer), 1, 14 (s, 18h), 1.48 (s, 18H), 2.68-3.12 (m, 8H), 3.30 (m, 2H), 4.08 (d, J = 9Hz) , 1H, second isomer), 4.10 (d,

J = 9 Hz, 1H, major isomer), 5.11 (d, J = 9 Hz, 2H).

Example 4

Preparation of intermediate H-Tbg-CH ₂ CH (CH ₂ C (O) CMe ₂ ) C (O) -Asp (cyPn) (Bzl) -NH- (S) -CH (CH ₂ C (O) MeJCH ₂ OBzl

The compound was prepared by the coupling reaction described in Example 1 from 3.42 g (7.13 mmol) of the title compound of Example 2 as the first reagent and 2.50 g (6.48 mmol) of the title compound of Example 3 as the second reagent. . The resulting crude compound was purified by flash chromatography after absorption onto silica. elution with 4: 1 hexane / ethyl acetate gave 4.64 g of the title compound N-Boc derivative in 84% yield (Rf = 0.21; hexane: ethyl acetate 7: 3). A solution of 4.64 g (5.44 mmol) of this derivative in 50 ml of a 6 N solution of hydrochloric acid in dioxane was stirred at room temperature for 1 hour and then concentrated under reduced pressure. The resulting residue was dissolved in ethyl ether and washed

A saturated aqueous solution of sodium bicarbonate, saturated with sodium fat, was dried over magnesium sulfate and concentrated to give a yellow oil containing two diastereomers. Flash chromatography on silica eluting with hexane / ethyl acetate. and methanol -V- ratio -5 4.5. ; -0.5 separates two .., -. ...

isomers, resulting isomer of higher polarity (Rf = 0.18;

- a mixture of cyclohexylation, hexane-methanol-4-monomer V-: a-z y

....____, Q, .5). colorless oil. ,with. yield.

In this example, the best-known compound of the invention is used without further purification. . -------- .-------- = The following example,

Example 5

Preparation of Boc- (N-Me) Val-Tbg-CH ₂ (R) CH (CH ₂ C (O) CMeJ C (O) -Asp (cyPn) (Bzl) -NH- (S) -CH (CH ₂ CMe) .) CH ₂ 0Bzl

The resulting compound was prepared by the coupling reaction described in Example 1 from 4.45 g (5.95 mmol) of the title compound of Example 4 as the first reagent and _from 4.41 g (17.9 mmol) of N-methylvaline as the second reagent. reagents. Flash chromatography of the crude product on silica eluting with hexane / ethyl acetate in a ratio? 3 'arises · 4-, 02 -g-resultant ·

SI < tb &gt;&lt; tb &^gt;&lt; tb &gt; Votti &lt; tb &gt; J &lt; 4 &gt; A &lt; 1 &gt; H, NMR (CDCl3): [delta] 0.87 (d, J = 7 Hz, 6H), 0, 90 (s,

18H), 1.10 (s, 9H), 1.48 (s, 9H), 1.5-5.0 (m, 10H).

......... 2.30 (m, 1H), 2.5-3.1 (m, 5H), 2.80 (s, 3H), 3.30 (m,

2H), 4.0 (d, J = 12.5 Hz, 1H), 4.20 (m, 1H), 4.32 (d, H = 8.5 Hz, 1H), 4.49 (d, J = 4.5Hz, 2H), 4.64 (d, J = 11Hz, 1H), 5.18 (d, J = 5Hz, 2H), 6.78 (br d, J =

8.5 Hz, 1H), 7.11 (broad d, J = 8.5 Hz, 1H), 7.1-8 (br. D, J = 11 Hz, 1H), 7.2-7.45 (m, 10 H).

Example 6

-P-ip-r-ava-PhCHgCHgCTOWN-MefVarl-Tbg-CHi-fEtt- -CH (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -yMeLeucinol

2025

A solution of 4.02 (4.25 mmol) of the title compound of Example 5 in 30 ml of a 6N solution of hydrochloric acid in dioxane was stirred at room temperature for 1 hour, and then stirred at room temperature for 1 hour. ηόν'.Λ ř-.Ί "Λ V" * * i -'T Τ '? yh Ί In T 1 7Ί ob O The N-terminal amino derivative of the title compound of Example 5 as its hydrochloride. After the coupling reaction described in Example 1 using this derivative as the first reagent and 2.00 g (13.3 mmol) of benzenepropionic acid as the second reagent, the crude product was purified after absorption on silica by flash chromatography eluting with hexane / ethyl acetate 3: 2 to give 4.00 g of white foam PhCH, CH ₂ C (O) - (N-Me) Val-Tbg-CH ₂ - (R) -CH (CH.C (O) -CMe ₃ ) C ( O) -Asp (cyPn) (OBzl) -NH- (S) -CH (CH ₂ CMe ₃ ) CH.OBz 1 (Rf = 0.35; hexane / ethyl acetate 1: 1) in 94% yield.

In the presence of 200 mg of 20% palladium hydroxide on charcoal, 4.00 g (4.03 mmol) of this compound are hydrogenolysed in an ethanolic solution at 50 psi hydrogen for 5 hours. After completion of the reaction, the reaction mixture was filtered through a 45 gm membrane.

The filtrate was concentrated under reduced pressure to give a clear oil which was then dissolved in 100 mL of ethyl ether. The solution was evaporated to dryness under reduced pressure. Dissolution and evaporation were repeated until a white solid formed. This material was triturated with hexane, filtered and dried under reduced pressure to give 3.12 g of the title compound in a yield of 95%.

^X H-NMR (d _s -DMSO), 400MHz: Compound in DMSO is a mixture of two rotamers in a ratio of 50: 50th

Δ 0.71-C5 H5 L5H), 1.06 (s,

4.5H), 1.20-1.78 (m, 10H), 1.93-2.16 (m, 2H), 2.48-2.88 (m, 6H), 2.84 (s, 1.5H), 2.92 (m, 1.5H), 2.96 3.06 (m, 1H), 3.10-3.23 (m, 2H), .delta. 4 -14 ^ (m-H) _{t. ±} 4 _r 22, _I (d, ^H_ = _± ,

4.62 (broad m, 1H), 4.73-4.81 (m (m, 6rf, 1H, tG., .Alpha .-) - ± -S-t, xy; ____0.5H) -, 8, 32 (d, J = 8.5 Hz, 5 Hz).

-i -Ϊ J-i

J, ií

3,

Hz, 0.5H), 4.54, 1.5 Η), 7.12 to 7.29 '''' essentially by Procedures ^R Ii guided "in the P * s object 6 'and to be only ϊ t1' 2- ( (PhCH ₂ ) ₂ CHC (O) - (N-Me) Val-Tbg-CH ₂ - (R) -CH (CH.C (O) CMe) Prepared Phenylmethyl) -3-phenylpropionic (dibenzylacetic acid) instead of benzenpropionic acid ₂ ) C (O) -Asp (cyPn) -MeLeucinol.

and

Example 7

Preparation _F Et ChNHC (O) _-Tbg-CH2 - (R) -CH (CH ₂ C (O) CMe,) CO-Asp (cyPn) -yMeLeucinolu ______

To a solution of the title compound of Example 4 (23 mg, 0.036) and 23 mg (0.057 mmol) of triethylamine in anhydrous methylene chloride was added 1-ethylpropyl isocyanate (28 mg, 0.248 mmol). The reaction mixture was stirred at 0 ° C for 1 hour under argon, followed by 18 hours at room temperature. The completion of the reaction is demonstrated>

thin layer chromatography eluting with a mixture of ethyl acetate: hexane (1: 1).

The residue was purified by flash chromatography after absorption on silica eluting with hexane / ethyl acetate (6: 4) to give 16 mg of the title compound dibenzyl derivative. ¹ H NMR (400 MHz, CDCl 3): δ 0.9 (t, J = 7 Hz, 3H),

0.92 (t, J = 7Hz, 3H), 0.94 (s, 9H), 0.95 (s, 9H),

1.10 (s, 9H), 1.25-1.90 (m, 14H), 2.52 (m, 1H),

2.68 3.27 (m-1-H -) - 2 -. - 3 - 1 - (- m - 1H), 2.94 - (dd, J = 7.2, 9 Hz, 1 H), 3.36 _3 ^ Q7__im ^ _2H -) _, - (dd, J = 5.5, 9 Hz, 1H), 3.46 (m, IH), 4.07 (d, J = 9) Hz, 1 H), 4.23 (m, 1 H) 4.28 (d, J = 9Hz, 1H), 4.48 (dd, J = 10Hz, 2H) 4, 66 (d, J = 10Hz, 1H), 4.74 (d, J = 9 Hz, 1 H), 5.17 (dd, J = 14Hz) , 2H) , 7.10 (d, J = 8.5 Hz, 1H), 7.2 -  /, 45 (111, xxi / '.' - ----------------- -------........

Hydrogenolysis of this compound at 100 psi in an ethanol solution in the presence of 10% palladium on charcoal gave the title compound. Mass spectrum: 703 (M + Na) +.

Example 8

Preparation of other intermediates for the preparation of C-terminal radicals of the peptides of formula 1:

Part a) _g NH - (R) -CH (Et) CMe _r

To a cooled solution of 0 ° C, 106 g (0.92 mmol) of 4,4-dimethyl-3-pentanone and 111 g (0.92 mmol) of (R) -α-methylbenzylamine vil benzene was added 50.5 ml ( 0.46 mmol) of titanium tetrachloride in 200 ml of benzene such that the temperature of the mixture did not rise above 10 ° C during the addition. The mixture was stirred at 40 ° C for 3 hours, cooled to room temperature, and filtered through diatomaceous earth, which was then washed with ethyl ether. The combined filtrate and washings were concentrated and the resulting residue was dissolved in 2 L of anhydrous methanol. The solution is cooled to 0 ° C and then 20 g (0.53 mmol) of sodium borohydride are added in portions so that the temperature of the mixture does not rise above 5 ° C. After evaporation of the methanol, the resulting residue was dissolved in ethyl ether and the solution was washed with saturated sodium chloride solution, dried over

This was evaporated to dryness to give the title compound

The oil was purified by flash chromatography on silica, eluting with ethyl acetate / hexane (7: 93) to give ~ 10-g * liquid-N = HT (-RT-phenethyl) - (R, R) ethyl-n-2. Of 2-dimethylpropylamine in yield. 54%. this liquid is dissolved in T / hexanes and 90 ml of a 6N acid solution of hydrogen chloride in dioxane are added to the resulting solution over 15 minutes. The foam was filtered and washed with hexane to give (TT5 (phenylethyl) -1- (R) -ethyl-2,2-dimethylpropyl hydrochloride).

¹ H NMR (CDCl 3):

δ 0.55 (t, J = 7.5 Hz, 3H),

1.14 (m, 8H) 15 Dec 3H), 2,36 - 7.48 (m, 4H) Solution 41.5

The mixture was stirred with a 10% palladium-on-charcoal solution and stirred in a Pan hydrogenation apparatus for 48 hours at room temperature and a hydrogen pressure of 50 psi. The resulting mixture was filtered and the filtrate was concentrated to give 25 g of the addition of hydrogen chloride, the title compound NH ₂ -R 1 -CH (Et) 2 - as a white solid in 100% yield.

1 H NMR (CDCl ₃ ): δ 1.10 (s, 9H), 1.22 (t, J = 7 Hz, 2H), 1.58-1.90 (m, 2H), 2.70-2 85 (m, 1 H),

8.10-8.40 (br-s, 3H)

Following the same procedure, but using 3,3-dimethyl-3-butanone instead of 4,4-dimethyl-3-pentanone,

NH- (R) -CH (Me) CMe ₃ . HCl.

Example 9

The preparation of the three cl-limit products for the preparation of the N-terminus of the peptides of the general formula 1 according to the procedure of Example 7 is as follows:

Part a): 1-propylbutyl isocyanate

This compound is prepared from the conventional πλivedο iivedPUPhf!

V.S.Goldesmidt and M.Wick in Liebigs Ann. Chem., 575, 217 (1952).

Part b): 1-ethyl-1- (2-propenyl) -3-butenylisocyanate

To 880 mL of a 1.0 M solution of allyl bromide in ethyl ether was added dropwise a solution of 14.5 g (264 mmol) of propionitrile in 40 mL of anhydrous ethyl ether. The reaction mixture was refluxed and stirred with a mechanical stirrer for 2 hours, and then cooled to 0 ° C. 320 ml of saturated aqueous ammonium chloride solution are carefully added to the cooled mixture. The organic layer was separated, dried over magnesium sulphate, cooled to 0 ° C and treated at the same temperature with 200 ml of a 1 M solution of hydrochloric acid in ethyl ether. The resulting solid was collected and dried under reduced pressure (about 27 g). The resulting product was dissolved in 200 ml of methylene chloride and the solution was washed twice with 10% aqueous sodium carbonate solution and then with saturated sodium chloride solution. After drying over magnesium sulphate and concentrating to dryness, a yellow oil is obtained which is distilled at 82-85 ° C and 52 kPa to give 11.06 g of a colorless liquid of 1-ethyl-1- (2-propenyl) -3-butenylamine with a yield of 34%.

¹ H NMR (400 MHz, CDCl ₃ ) of this compound: δ 0.89 (t, J =

Hz, 3H), 1.39 (q, J = 7Hz, 2H), 2.11 (d, J = 7Hz,

2H), 5.06-5.14 (m, 4H), 5.80-5.89 (m, 2H).

The above compound was converted to 1-ethyl-i- (2ιτορ-erryt) ^- -3-drums L-isojcyaj; iát_jvý.še__uyed.enýiiL_.pc5bupem · by VSGoldsmidta and M. Wick,

First part of the previous part b

-fttvl - ** j -

-1- (2-propenyl-3-butenylamine), starting from the method described

By G. Alvernh and A. Laurent in Tetrahedron Lett., 1057 (July 1, 2010). procedure, using.

Other reactive reagents with unsaturated, unsaturated substitution can be prepared, i.e., unsaturated alkylaminocarbonyls such as 1-methyl-1- (2). (propenyl) -3-butenyl. Note that if isocyanate intermediates prepared according to the procedure of Example 7 are needed, the final product will correspond to the peptide derivative of Formula 1 having the N-terminus saturated. In addition, according to the procedure of Example 7, the corresponding peptides such as Pr ₂ CHNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) -C (O) - Asp (cyPn) -NH- (R) -CH (Et) CMe ₃ , FAS mass spectrum (m / z): [M + H] ⁺ - the resulting peptide derivative mentioned in Example 10.

Using suitable intermediates, the compounds of formula (I), some of which are listed in the table of the following example, can be prepared according to the procedures of Examples 1 'to 7'. No precipitate is formed. In some cases, the compound can be purified by reverse phase semi-preparative HPLC on a C-18 column, eluting with a gradient of acetonitrile and water containing a 0.06% trifluoroacetic acid solution. In this case, before purification, the crude product is dissolved in 0.1 M aqueous ammonium hydroxide solution and converted to pH = 7 by adding 0.1

M aqueous acetic acid solution. If appropriate, the diastereomeric mixtures are separated as described above.

Examples of other compounds of formula 1 that can be prepared as described herein include (PhCK ₂ ) g CHC (O) (N-Me) Val-Tbg-CH ₂ - (R) ₁ CH (CH ₂ C (O) CMe _g ) C (O) -Asp (cyPn) -NHCH ₂ CMe ₃ and (PhCH ₂ ) ₂ CHC (O) - (N-Me) Val-Tbg-CH ₂ (R) -CH (CH ₂ C (0) CMe ₃ ) C (O) -Anf Λ ,, Ο / r '. Μλ λ '· ·' - ₂ ·

Further examples of compounds are provided below. 1:

Pr ₂ CHNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -NHCH ₂ CMe FAB (mass spectrum m / z: 665) M + H / ⁺

EtPr ₂ CNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -NH- (R) -CH (Et) CMe ₃ ,

FAB / MS m / z 722 (M + H) &lt; + &^gt;

E tP r ₂ CNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPh) -NHCK ₂ CMe ₃ ,

FAB / MS m / z 693 (M + H) @ ⁺ .

Example 10

Experiments in which the effect of a peptide derivative of the formula I according to the invention was investigated,

Pr ₂ CHNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) -CMe ₃ ) -C (O) -Asp (cyPn) -NH- (R) -CH (Et) CMe ₃ , and a combination of this agent with acyclovir against a wild type HSV-1 clinical isolate and two acyclovir resistant clinical isolates HSV-1.

AND

- 38 Viruses

Clinical isolates of bvlv are described in S.L. Sacks et al, Annals of Internal Medicine, - · 111, 893, 1989. Iso ⁼ methacrylates 'were "zísxány ^ OA' in 2ény ^ ⁼ ^ 25 ^ age et" 5 ^ & kutrrí niyeiogen ^ ~ ^ her leukemia. during the first relaxation of the allogeneic bone marrow transplant, the initial culture of the virus was excreted from the material taken from the eighth. days after transplantation. .. 'cone marrow. Positive (isolate 294 defined as but the viral cultures were still positive at day 36 (isolate 615, an HSV-1 that is already resistant to acyclovir). It has also been shown that subisolate 615.9 (ACV TK) Later, it was shown that subisolate 615.9 was a TK-deficient mutant, and in contrast to this isolate, subisolate 615.8 (ACV POL) was shown to be acyclovir resistant to both acyclovir and forcarnet. forcanet, but the thymidine kinase activity is close to that shown for the original isolate Qd Snirinate siE-fl or as far as delayed

«.V v - _ _ _ _. _ v ~ r u ---- ~ ~ w ·

DNA polymerases.

Performing the test

BHK-21 / C13 cells were incubated for two days in T-flasks on a 150 cn area (1.5 x 10 cells / flask) using alpha-MEM medium (Gibco Canada Inc, Burlington, Ontario, Canada) supplemented with 8% by volume fetal calf serum (FBS,

Gí bco? 0 requests'! well? ) T "Mater Buhěchý ^ ^r IAR" albeit "in wild tTpů sobe'rrxm ^ trypsin and then the cells were transferred to fresh culture medium to 24-well plates in each well was 2.5 x 10 ^ cells in 750 .mu.l of culture medium . The cell material was incubated for 6 hours at 37 ° C so that the cells could adhere to the support. Then, the cell material was washed once with 500 microliters of medium and 0.6% by volume FBS and then incubated in 750 microliters of the same low serum medium for a total of 3 days. After this time, the low serum medium is removed and the cell material is incubated in 500 microliters of BBMT medium for a total of 2-3 hours. The BBMT environment is described1 “- ''« <* <? !! $ 'η —λ «« ι

USA, 79, 7909, 1982. The cells are then infected with HSV-2 using 0.02 PFU / cell in 100 microliters of BBMT medium. The HSV-2 strain used was the HG-52 strain according to Y. Larigelier and G. Buttin, J. Gen. Virol., 57, 21, 1981, the virus was stored at -80 ° C. After one hour of adsorption of the virus at 37 ° C, the medium was removed and the cell material was washed with x 250 microliters of BBMT. The cell material in each well was incubated in the presence or absence (control) of a certain concentration of the test substance dissolved in 200 microliters of BBMT medium. After incubation for 29 hours at 0 ° C, infected cells were separated by freezing the plate at -80 ° C followed by thawing. Cell material from each well was separated from the surface by melting ice fragments. After complete thawing, the cell suspension was separated and each well was rinsed with 150 microliters of BBMT medium. The virus sample (suspension and washing liquid) was cautiously sonicated for 4 minutes at 4 ° C. The cell debris was collected by centrifugation (1000 g, 10 min at ° C). The supernatant was collected and stored at -80 ° C until virus titer was determined.

Virus titration was performed using a colorimetric assay modification according to M. Langlois et al., Journal of Biological Standardization, 14, 201, 1986.

In a similar manner to the above, BHK-21 / C13 cellular material is trypsinized and transferred to fresh medium in a 96-well microtiter plate, one

----- 40 · = wells contain 20,000 cells in 100 microliters of culture medium. The cell material was incubated at 37 ° C for 2 hours. During this time, the virus sample was thawed and fractionated by sonication for a total of 15 seconds, and then the logarithmic dilutions of the sample were prepared.

- · liter sample 200..mikrolitrů BBMT medium, sequential dilutions were held vícekánáíovou pipětóujv '- - - ^ incubation cell function ^ ^ ^ BNK -aiÁClO ^^^ ^ p ~ h 2ivňe přosťře'dl ⁼ Ti ^{- II} ^ ahraďí by means of alpha-MEM supplemented with 4% v / v FBS. Now the cell material can be infected with different virus samples at different dilutions. Aliquots of various dilutions of 50 microliters are transferred to the appropriate plate wells. The resulting infected cells were incubated at 37 ° C for two days. Then 50 microliters of a solution containing 0.15% neutral red (in% by volume) in Hank's balanced salt solution (pH 7.3, Gibco Canada Inc) is added to each well. The plate thus prepared is incubated for 45 minutes at 37 ° C. The culture medium is removed from each well and the cell material is washed with 200 microliters of Hank's balanced salt solution. After washing, the dye is released from the cells by addition of 100 microliters of a 0.1 M Soren-Esenate Buffer Buffer in an amount of 1-1. _

Sorencen citrate buffer was prepared by first preparing a 0.1 M citric acid disodium salt solution by dissolving 21 g of citric acid monohydrate in 200 mL of IM. aqueous sodium hydroxide solution and sufficient water was added to 1 liter. 61.2 ml of a 0.1 M solution of sodium tartaric acid were then mixed with the water. The hydrochloric acid solution was adjusted to pH 4.2, if necessary. The mixture in the wells is then mixed gently with a stirrer and the wells are read using a spectrophotometer at 540 nm to determine the number of viable cells. In this way, it is possible to determine the inhibition of viral growth in percent for various concentrations of the test substance and to calculate a value of ΙΟ ^ θ, that is, the concentration of the test substance which causes 50% inhibition of virus replication.

This procedure was used to evaluate acyclovir and the peptide derivative individually and then in various combinations. Therefore, the efficacy of the two compounds against the individual isolates was demonstrated. In addition, the synergistic effect of both compounds could be evaluated and confirmed using the isobol method to evaluate the results obtained in these assays, as described in J. Suhnel, J. Antiviral Research, 13, 23, 1990.

With regard to the isobol method, this method requires experimental data from two test substances to be eaten sensitively and in different dose combinations with approximately the same level of efficacy. Selected concentrations of peptide derivative _? (IC ^Ic I0 '- ^ 20 ^and ICso) therefore combined with various DAV * · ..kami acyclovir was measured and found ICCR .. putting a lot _t> for the peptide derivative and the same values for acyclovir were derived from curves obtained in preliminary experiments.

Isobologram can be created using the value marked

FICgQ (acyclovir), is the ratio of acyclovir concentration required to inhibit HSV replication by 60% in the presence of a certain concentration of peptide derivative to that required in the absence of the peptide derivative.

*. This value is plotted against a value that is a ratio of a fixed concentration of peptide derivative to that concentration of peptide derivative that causes 60% inhibition of HSV replication in the absence of acyclovir. The following equations can be used to calculate:

AND

X-axis:

- 42 fixed concentration of peptide derivative

IC p r o_p ep class_jd_e r i weight

ICC ', - (acyclovir * X-micromol peptides 60 * /

FIC _en (acyclovir)

IC _An for 'acybtovirus alone'—

Results:

Table 1 shows the results obtained when acyclovir and the peptide derivative were examined individually using a wild-type HSV-1 clinical isolate and the two acyclovir resistant clinical isolates HSV-1 mentioned above.

__________Table I compound isolate

IC. «

294 isolate 615.8 _{/ U} M IC ₅₀ / UM isolate 615.9 ^IC ₅₀ 'acyclovir. peptide derivative

-2.0

6.2 The results of Table I show that the compounds of formula 1 are active against wild type HSV-1 and have similar activity against acyclovir-resistant HSV-1, for example against POL-mutants and TK-deficient mutants, where acyclovir loses much of its effectiveness and is against it

- these mutated strains are 20x to 40x less effective.

The following Tables II, III and IV show: - bvi in the combination of acvlo virus and peptide derivative against the clinical isolate of wild-type HSV-1 and the two above-mentioned clinical isolates of HSV-1, resistant to acyclovir.

Table II '^; Antiviral activity - tests for synergistic effect

Clinical isolate 294 fixed concentration of peptide derivative (u µM) acyclovir IC ^ ¹ in combination with peptide derivative (UM)

0.3 0.85 0.5 0.70 0.7 0.35 0.9 0.30 1.0 0.30 1.2 ’ 0.22 1.4 0.10 1.6 0.13

= 44 _in -

T-α-b-xr-t-kProtiviral activity - assays for synergistic effect.

X X á ~ 6 15 15 15 15 15 15 15 15 15 AG AG AG AG AG AG y y y y y y y y y y y

„5. _ a ~ - ^ t-4- ·· Jx.rAR .- *

9. ^Λ »Λ \ ην | Γ Tf '. ,,' * in combination, 'Z- ..... -. . ·; with peptide derivative (γuM) »-Ak3» _

0.5

0.7

0.9

1.0

1,2

1.4

1.6 i5; o *

1370 ’

7.0

8.0

5.3 5.0 1.7

2.4

Table IV

Antiviral activity - tests for synergistic effect

Isolate 615.9

AO and X _ _{=, r,} _ a fixed peptide concentration derivátu- (.mu.M) θ ^ ΙΟ acyclovir in combination with a peptide derivátem'- (yum) 0 '33TČT ~ "~ L_

17.0

5.5

5.6 2.0 0.9 £ 0.02 <0.02

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

Comment:

Values ΙΟ, -θ are not corrected for solubility.

All stock solutions were centrifuged in an ultra centrifuge at 100,000 g for 30 minutes at 4 ° C before use.

The solubility of the peptide derivative at a concentration of 20 micromoles is 85%.

The cytotoxicity of the peptide derivative is 89 micromolar (value is shown without correction).

1) IC 50 values obtained from a concentration-response curve for acyclovir ranging from 7.6 x 10 ^-4 to

1.5 x 10 .mu.mol in the presence of a fixed concentration of the peptide derivative in the left column.

2) IC 50 values obtained from the response curve

- -4 2 dose for acyclovir ranging from 1.5 x 10 to 1.5 x 10 micromoles in the presence of a fixed concentration of the peptide derivative in the left column.

The results shown in Tables II, III and IV show that the peptides of Formula 1 can potentiate acyclovir activity against wild type HSV-1 and acyclovir resistant HSV-1 mutants when both are used. in combination. The results show that there is a proportional decrease in the IC 50 value for acyclovir as the ratio of the concentration of the peptide derivative of the invention to acyclovir increases.

BRIEF DESCRIPTION OF THE DRAWINGS

Ρ »Fig. 1 graphically represented 1sou positive results, demonstrating the synergistic effect of the substances used and the obtained appli ^zi ^ ff ^ S ^^ ⁱ ^ ^^^ him R¹ = F! <^ Škach; siaoyoláviren _{l ;;;} and a peptide derivative against a clinical SIS sub-isolate. 8 ........

(ACV ^r POL) ~. . .. ..........

2 ', positive results are shown, acyclovir was used in combination with a peptide derivative against the clinical subisolate 615.9 (ACV ^r TK).

Represented by:

Ο <J Γ “70>

C · ζ '3 *. <' ./·> >> ο c Ό

1 * 0

C7 ο '* χ

X

C3

LD σ>

---, / w ?? - i

OC

Claims (16)

Peptide Derivatives of Formula 1 A-3-D-CH 2 .CK / CW., C (O) 2 / O (0) -NKC 1; CR ² (R ³ ) COOH / C (O) -1 (1) where A is phenylacetyl, phenylpropionyl, (4-aminophenes Doropionyl, (4-fluorophenyl) propionyl, (4-hydroxyphenyl) propionyl, (4-methoxyphenyl) propionyl, 2- (phenylmethyl) -3- * phenylpropionyl, 2- 1- (4-fluorophenyl) methyl] -3- (4-fluorophenyl) propionyl, 2 - [(4-methoxyphenyl) methyl] -3- (4-methoxyphenyl) propionyl, or benzylaminocarbonyl, B represents (N-Me) -Val or (N-Me) -Ile or forms together A and B a saturated alkylaminocarbonyl group, butylaminocarbonyl, 1-methylehylaminocarbonyl, 1-methylpropylaminocarbonyl, 1-ethylpropylaminocarbonyl, 1,1-dimethylbutylaminocarbonyl, 1-ethylbutylaminocarbonyl, 1-propylbutylaminocarbonyl, 1-propylbutylaminocarbonyl, 1-methylbutylaminocarbonyl, 1-butylpentylaminocarbonyl, 1-ethylbutylaminocarbonyl, 2-ethylpentylaminocarbonyl, 1-methyl-1-propylbutylaminocarbnyl, 1-ethyl-1-propylbutylaminocarbonyl, 1,1-dipropylbutylaminocarbonyl, (1-propylcyclopentyL) aminocarbonyl or (1-propylcyclohexyl) aminocarbonyl, D is Val, Ile or Tbg, R ¹ is 1-methylethyl, 1,1-dimethylethyl, 1-methylpropyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclooentyl, cyclohexyl, 1-methylcyclopentyl, a group NR ⁴ R ⁵ wherein R ⁴ represents a hydrogen atom or a lower alkyl a - R represents lower alkyl or forms R and R together with the nitrogen atom to which they are attached a pyrrolidine, piperidine, morpholine or 4-methylpiperazine ring, AND - 48 R ‘denotes a hydrogen atom and Sx denotes methyl, ethyl, 1-methylethyl, 1,1,1-methyl-propyl, 2-propyl or benzyl, wherein the? In addition, the groups R @ 1 are bound and R @ ³ has the configuration R @ 1 or - _and - and is not dependent Ie it bears methyl-1-th e-yl-1- naho.- t.vo rx sp.o ___ greedily · -s-atomea. .uhlíku, Ji they are attached form a cyclobutyl, cyclohexyl or _ evkiešehtvV and NKR represents ^S, wherein R ⁹ is 2-methylpropy1, 2,2-dimethylpropyl. 1 (R), 2,2-trimethylpropyl, 1,1,2,2-tetylpentyl oil 1,1,1 (R) -ethyl 1-2,2-dimethypropyl, 2- (R, S) -methylsulfonyl, 2,2-dimethylbutyl, 3,2-dimethylbutyl, 1 (R), 2,2-trimethylsulfonyl, 1 (R), 3,3-trimethylbutyl, 2-ethylbutyl, 2,2-methylbutyl, 2-ethyl-1H-methylbutyl, 2-ethyl-2-methylbutyl, 1 (R) -ethyl-3,3-dimethylbutyl, 2,2-dimethylpentyl, cis- or trans-2- methylcyclohexyl, 2,2-dimethylcyclohexyl or cyclohexylmethyl; or represents a radical NHGH Ξ (R the carbon atom to which is bonded group R 'has the configuration 5, R ^z is i, 1-dimethylethyl L-mezhyl? rspy1 2-methy1p ropy1, 2,2-dimethylpropyl or cyclohexyl. 2-methyl, a. 2 is CK-GK, C (O) OK, C (O) 1, 3 '' 3 '' or 3 (0) 02, wherein R ³ is methyl, ethyl or propyl as well as pharmaceutically acceptable salts of these compounds, for use in the treatment of acyclovir-resistant herpes simplex infections. Peptide derivatives of the general formula I according to claim 1, in which A represents phenylpropionyl, 2- (phenylmethyl) -3-phenylproponylonyl or benzylaminocarbonyl, -3- (N-Me) Val, 0 stands for 7b g, R ¹ is X-methylethyl _f 1,1-dimethylethyl, 1-methylpropyl, "» n »e» Ow / «<ς i -""_W .. y. C 1-4 alkyl, cyclopentyl, cyclohexyl, or 1-methylcyclopentyl, 3 represents a hydrogen atom; R is methyl, ethyl, 1-methylethyl, propyl, or benzyl, wherein the carbon atom to which the groups are attached is 3 R and R has the R configuration; R @ 1 and R ^{@ 2} independently represent methyl or ethyl or form together with the carbon atom to which they are attached cyclobutyl, cyclopentyl or cyclohexyl and Ξ represents a group ΝΚΗθ, where R 6 represents 2,2-dimethylpropyl; 1 (R), 2,2-trimethylpropyl, 1 (R) -ethyl-2,2-dimethylpropyl, 2,2-dimethylutyl or 1 (R) -ethyl-3,3-dimethylbutyl or is znamená NHCK (R ^/) -Z wherein the carbon atom to which the R group is bound has an S configuration, R ⁷ represents 2,2-dimethyloropropyl and Z represents CK-OK, C (O) OK, and C (O) NH ₂ or C (C) OR, wherein R is methyl, ethyl or propyl, as well as pharmaceutically acceptable salts thereof, for use in the treatment of acyclovir-resistant herpes simplex infections. AND - SO 3. Peptide derivatives according to claim 1, of the general formula wherein A and S are taken together to form a saturated alkylaminocarboxylic acid selected from the group consisting of C1-4-thiol-4-carboxylic acid and phenylcarbonyl-1,1-propylthutylaminocarbonyl. R-ethylpentylaminocarbonyl! _5-1-L-meShyl propylbutylam'inb'karbdhyl l-Ethyl-t-supersaturate. ~ '· BUtylamiňakarbonyl --1 <L diaropylbutylaminpkarbany! and cygicycyclopentyl <mincicarbonyl, __ 1. _ 0 means Tbg, R 1 represents 1-methylethyl 1,1,1-dimethylethyl, 1-methylpropyl, 1,1-dimethylphenyl, 2,2-dimethylpropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 1-methylcyclocentyl, R is hydrogen and R ^ represents methyl, ethyl, 1-methylethyl, prooyl or benzyl, the carbon atom to which the groups are attached ^; * 3 R 'a and has an R configuration, or? 3 R and R independently represent methyl or ethyl or form ____________ together with the carbon atom to which they are attached a cyclo butyl ........ ,, .cyklopentyL or cyklohexyKaT ^---: £ NKR represents a group wherein R is 2,2 (petroleum), (KR), 2,2-trimethyloropropyl, (KR) -ethyl-2,2-dimethyl. propyl, 2,2-dimethylbutyl or 1 (R) -ethyl-3,2-dimethylbutyl; represents S or a group NHCH ^(R7) -Z '' wherein the carbon "acWrnarhě / SW ^ e ^ linked-acoust-ina-R ^ -has konflguracl JS R ⁷ is 2,2-dimethylpropyl; and 2 is CH 2 OK, C (Q) 0K, C (O) NH ³ , or C (O) OR ³ , wherein R ³ is methyl, ethyl; or propyl, as well as pharmaceutically acceptable salts of these compounds, for use in the treatment of acyclovir resistant herpes simplex infections. Peptide derivatives of the general formula 1 according to claim 1 selected from the group: PhCH ₂ CH ₂ C (O) - (N-MeVai-Tbg-CHg- (R) -CH (CHgC (O) -CMe ₃ ) C (O) -Asp (cyPn) -gammaMeLeucinoL " CHtgCHNHC (0) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (O) -As? (CyPn) -gammaMeLeucinol, Pr ₂ CHNHC (O) -Tbg-CH ₂ - (R) -CH- (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -NK- (R) -CH (Ht) CMe ₃ , Pr-CHNHC (O) -Tbg-CH, - (R) -CH (CHC (O) CMe-) C (O) -As ₂ (cyPn) -NHCH ₂ CMe ₃ , ₂ t r ₂ CNHC (O) -T 3 g-CH ₂ - (R) -CH (CK ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -NH- (R) -CH (Zt) CMe a P tP ₂ CNHC (O) -Tbg-CH _? (R) -CH (CH 3 C (O) CMe) C (O) -As ₂ (cyPn) -NHCH ₂ CMe ₃ , as well as a pharmaceutically acceptable salt thereof, for use in the treatment of acyclovir-resistant herpes simplex infections. - 52 5. Use of the peptide derivatives of the general formula I according to claim 1: *, - 3-0 ^ CH 2 CH / CH 2 C (Q) S. ¹ / C (O) -NHCK / CR ² (R ³ ) CQOK / C (O) -E (i) 'where *'' - - - - - - - - _______ ... _________ A ⁼ ^ znam means acac © ty ifM if if ifionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionionion (4-fluorophenyl) propionyl 4-methoxyphenyl) proplenyl, 2- (phenylmethyl) -2-phenylpropionyl, 2 - [(4-fluorophenyl) methyl] -2- (4-fluorophenyl) propionyl 1,2-f (4-methoxyphenyl) methyl -3- (4-methoxyphenylpropionyl or benzylaminocarbonyl), and is (N-Me) -Vai or (N-Me) -lle or together form A and 3 saturated alkylaminocarbonyl groups are butylaminocarbonyl, 1-methylethylaminocarbonyl, 1-methylpropylasinocarbonyl, 1-ethylpropylaminocarbonyl, 1,1-dimethylbutylaminocarbonyl, L-ethylbutylaminocarbonyl, 1-propylbutylaminocarbonyl-1-propylbutylamine. ---- Bu-butylene amine. Cocbonyl 1-ethylbutylaminocarbonyl, 2'-ethyl-p-ethylaminocarbonyl> 1-methyl-1-propylbutylaminocarbonyl, I- ethyl-1-propylbutylaminocarhonyl, 1,1-dipropylbutylaminocarbonyl, (1-propylcyclopentyl) aminocarbonyl or (1-procylcyclohexyl) aminocarbonyl, D is Val, Is or Tbg, 6 is 1-methylethyl, 1,1-dimethyisthyl, 1-methylpropyl, 1, L-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclooentyl, cyclohexyl, 1-methylcyclopentyl, group 4 5 4 NR R, wherein R is hydrogen or lower alkyl; 3 at 5 - ~ R is lower alkyl or forms. R and R together with the nitrogen atom to which they are attached a pyrrolidine, piperidine, morpholino or 4-methylpiperazine ring, And R 'is methyl, ethyl, 1-methylethyl, 1,1-dimethylethyl, propyl, 2-propenyl or benzyl, wherein the carbon atom to which R' and R are bonded ^ has the configuration R, or Leona with the carbon atom to which they are attached cyclobutyl, cyclopentyl 'or cyclohexyl, and 5 5. '. £ represents an NHR group wherein R represents 2-methyxpropyx, 2,2-dimethylpropyl, 1 (R), 2,2-trimethylpropyl, 1,1,2,2-tetramethylprooyl, 1 (R) -ethyl-2,2-dimethylpropyl, 2- (R, S) -methylbutyl, 2,2-Dimethyl-butyl, 2,3-dimethylbutyl, L (R), 2,2-trimethylcutyl, 1 (R), 3,2-trimethylcutyl, 2-ethyl-butyl, 2,2-diechylbutyl, 2- ethyl-1 (R) -methylbutyl, 2-ethyl-2-mechylcyl, (1R) -ethyl-2,2-dimethylbutyl, 2,2-dimethylpertyl, cis- or trans-2-methylcyclohexyl, 2,2 -dimethyloyclohexyl or cyclohexylmethyl or is NH NHCH (R) -Z, wherein the carbon atom to which R is attached has the configuration Ξ, R? means 1,1-dimethylethyl, 1-mechlorooyl, 2-mechyloropropyl, 2,2-dimethylpropyl or cyclohexylmethyl; and Z is CH-OK, C (O) OK, C (O) NH 3 9 or C (O) CR, wherein R is methyl, ethyl or propyl, or a pharmaceutically acceptable salt thereof for the manufacture of a pharmaceutical composition for treatment acyclovir-resistant herpes simplex virus infection in mammals. θ. Use of peptide derivatives of the general formula 1 in which: A is phenylpropionyl 2- (phenylmethyl) -3-phenyloropio; nonyl or benzylaminocarbonyl; 3 stands for (N-Me) Val, 0 means Thg, 1, l-dimethyl thyIpropyl @ 2 ₅ 2 ₅ ° dímethylproeyl cyclobutyl, ._ ..ycyx fc / _{C QúčÁy Ry .or 4 "IOE kiope.ii * y <· t and • a * represents a hydrogen atom and represents 'methyl IV' of methyl 1-methylethyl-1-or enyl, the carbon atom to which the groups 2 are bound R and R have the configuration R 1. or and R ³ independently represent methyl or ethyl or form together with the carbon atom to which they are attached a cyclobutyl, cyclopentyl or cyclohexyl group and represents an NHR ³ group wherein R 0 represents 2,2-dimethylpropyl, 1 (R), 2,2-trimethylpropyl, 1 (R) -ethyl-2,2-dimethynylpropyl, 2,2-dimethylbutyl or 1 (R) -ethyl-3,3-dimethylbutyl, or is fused NHCK (R R) - Z, wherein the carbon atom to which the R group is attached has the S configuration, and the ^7- substituted 2,2-dimethylamino-2-is -CH-OH-r -C-CO-) -OH, c (o ) NH, or- inu Grp C (O) oR ³ wherein R ³ represents methyl _l: _ ethyl or propyl. or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a herc-es virus simplex virus infection in a mammal. 7. Use of peptide derivatives of the general formula I in which they form a saturated saturated amine carbonyl of 1-ethyl-propyl-aminocarbonyl, L-ethyl-butylaminocarbonyl, 1-propyl-butylaminocarbonyl, 2-ethyl-pentylaminocarbonyl, i-methyl-i-propylbutylaminocarbonyl, 1-ethyl-L-propylbutylaminocarbonyl, 1,1-iiproputyutylaminocarbonyl; and (L-p Crude Oil CyclopentsUminocarbonylL, D stands for Tbg, 8 * is 1-methyl-ethyl, 1,1-dimethyl-ethyl, L-methyl-propyl L, L-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclopentyl, cyclohexyl, or 1-methylcyclopentyl, R @ 1 is hydrogen and R is methyl, ethyl, L-methylethyl, propyl or benzyl, the carbon atom to which the groups are attached 2 3 R and R have an R configuration, or 2 3 R 'and R' are independently methyl; or ethyl or together with the carbon atom to which they are attached cyclobutyl, cyclopentyl or cyclohexyl, and Ξ represents a NKR group wherein R is 2,2-dimethylpropyl, 1 (R), 2,2-trimethylpropyl, 1 (R) -ethyl-2,2-dimethylprooyl, 2,2-dimethylutyl; or L (R) -ethyl-3,3-dimethyl-octyl or NK is NKCH (R ⁷ ) -Z, wherein the carbon atom to which R ⁷ is attached has the configuration 2, R ⁷ is 2,2-dimethyl-propyl, and 2 is CH 3 O, C (O) OK, C (O) NH 3, or a C (O) QR 4 group, wherein R 6 is methyl, ethyl or propyl, or a pharmaceutically acceptable salt thereof. compounds for the manufacture of pharmaceutical compositions for the treatment of acyclovir resistant herpes simplex virus infections in mammals. Use according to claims 5 to 7, wherein the peptide derivative is selected from the group PhCH ₂ CH ₂ C (O) - (N-MeVal-Tbg-CH ₂ - (R) -CH (CH ₂ C (o) -UMe ₃ ) C (O) -Asp (cyPn) -gammaMeLeucinol, Et ₂ CHNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (O) -Asp (cyPn) -Lucin 1 '. ^'41 ~. ϊ,. ' ^K -o *. ' --- J ** · ♦ * «<“ - W f ~ Λ. ϋ ií * T.Λ ³ \ ** 2 * \ '·' · ~ '*''V - J - · * r ______-. NH * (.R.) - CH (Et) CMe _3. ,, , __________ _ ZZpSShIi) ^ TAs p (cy P rv-NHCH ^ CMe ^), E tPr ₂ .CNHC. (O) -TBg-CH ₂ - (R) -CH (CHgC (O) CMe ₃ ) C (O) -Asp (cyPn) -NH- (R) -CH (Et) CMe ₃ and EtPr ₂ CNHC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe) C (O) -Asp (cyPn) -NHCH ₂ CMe ₃ , Use of a peptide derivative according to claim 1, or a pharmaceutically acceptable salt thereof and a nucleoside of an antiviral effect, or a pharmaceutically acceptable salt thereof, in combination for the manufacture of a pharmaceutical composition for treating acyclovir-resistant herpes simplex virus infection in ssavs. Use of a combination according to claim 9, wherein the component of the combination is a peptide derivative of the general formula I, in ng 2 A is phenylpropionyl, 2- (phenylmethyl) -3-phenylpropionyl or benzylaminocarbonyl, B stands for (N-Me) Val, stands for Tbg, R is 1-methylethyl, 1,1-dimethylethyl, 1-methyleropropyl 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclooutyl, cyclopentyl, cyclohexyl, or 1-methylcyclopentyl, ~ 2 R is hydrogen and R is methyl, ethyl, 1-methylethyl, prvi or benzyl, wherein the carbon atom to which R ² and R ³ are attached is of the R configuration; or R ² and R ³ independently represent methyl or ethyl or form together with the carbon atom to which they are attached cyclobutyl, cyclopentyl or cyclohexyl; and δ represents NHR °, where R ° represents 2,2-dimethylpropyl, 1 (R), 2; , 2-trimethylpropyl, 1 (R) -ethyl-2,2-dimethylpropyl, 2,2-dimethylbutyl or I (R) -ethyl-3,3-dimethyl-7-butyl or is NH NHCH (R) -Z, where carbon 7 the atom to which the group R is attached and having the configuration 2, R ⁷ is 2,2-dimethylpropyl and Z is CH-OK, C (O) OK, o 3 C (O) _NH2 or C (O) OR where R is methyl, ethyl or propyl, c, or a pharmaceutically acceptable salt thereof. Use of a combination according to claim 9, wherein the component of the combination is a peptide derivative of the general formula I, in which A and 3 are taken together to form saturated alkylaminocarbonyl for the groups L-ezhylaaropylaminocaroline L, L-echylbutylaminocarboxylic acid, L-propyl acylaminocarboxylic acid, L-propyl amine, L-methyl amine, L-propylamine , 1, L-diaropyLbenzylaminocarbanyl and (1-propylcyclopenzyl) amino carbones L, - - = ~ 58 - = · - means Tbg, and ^L means 1-mechylechyl, 1Λ-dime chyle chyl, 1-methyl? 1,1,1-dimethylethyl, 1,2-dihydro-1,2,3-diol-2-ol. Crude oil, cyclobucyl, γ-hexyl-Fnl-g-methyl-cyclopentyl--Mi-ZS.iZ, i-OS-zinc-i-2. . .... ... .......... and ^3- denotes mechyl, ethyl, methyl, methyl, p-methyl or benzene. _ ^"= ^ ^T ^ * '^ ^and syrr ricémž * aC3m- ^with uhííkuT» on. _s ne jS- „j ow jásány _jkug iny ~ ———' —— .—- - --- ____.... ______ ---- '~ · 3. ~ a R ^J has a configuration ^ di ^ RT ^ or '^^ · --... and ² and ² independently represent methyl or echoes; or, together with the carbon atom to which they are attached, form a cyclopercyl; or cyclohexyl a S is the NKR group ^S , where? ⁵ is 2,2-dimethyl-petroleum, 1 (R), 2,2-dimethylethyl, 1 (R) -ethyl-2,2-dimethylpropyl, 2,2-dimethylbutyl or (a) -Ethyl-2,3-dimethylbutyl; or Z represents NKCHCR ⁷⁾ - !, where acoa carbon to which it is attached and group ⁷ has a configuration 5, A 'is Z, 2-dimethyl Chýle? oil! and Z is CH-OK, C (JGK), and 3 C (G) _NH2 sky C (O) O ', wherein A represents mosses !. echy! or propyl ___ _______ _L or its salts acceptable from pharmaceutical viewpoint. Use of the combination according to claim 9, wherein the component of the combination is a peptide derivative of the general formula I from the group - -------- PhCH ₂ CH ₂ C (O) - (N-MeVal-Tbg-CH ₂ - ( R) -CHCCH 2 C (O) -CMe ₃ ) C (O) -Asp (cypr.) - gammaMeLeucinol, Ξ C ₂ ChNHC (0) -THG _CK-2 - (A) -CH (CK ₂ C (0) CMe ₃₎ C (0) -As? (CyPn) -gammaMeLeuc ino1, ΐ ? R CHNHC (C) -Tbg-CK- (R) -CH- (CH ₂ C (O) CMe ₃ ) C (O) -As- (cylene) -NK- (R) -CH (Et) ) CMe ,, EXAMPLE ₂ ChNHC (O) -Tbg-CH - (R) CH _(CH? C (O) CMe _Ί) C (O) -As? (Cy? N) -NHCH CMe ^., E t r ₂ CNHC (O) -T 3g-Crt ₂ - (a) -CH (CK ₃ C (O) CMe 2) C (O) -As- (cylene) -NH- (R) -CH (Et) C14e- and xa>> E t R ₂ CNHC (O) -Tbg-C 1 - (R) -CH (CH-C (C) CMe) C (O) -As (cyPn) -NHCH-CMe; 2 3 · 13. Use of a combination according to claim 9, wherein the components of the combination nucleoside analogue of formula 2 ock ck _{2 2} c - ₂ ok wherein R ⁹ is hydrogen, hydroxy or amino, or a pharmaceutically acceptable salt thereof. Use of the combination according to claim 9, wherein the component of the combination is a nucleoside analogue selected from the group of vidarabine, idoxuridine, trifluoridine, ganciclovir, edoxudine, broavir, fiacitabine, penciclovir, famciclovir and rociclovir. Use of a combination according to claim 9, wherein the component of the combination is a peptide derivative 26 groups FhCH-CH-C (O) - (N-Me Val -Tbg-CH- -CR) -CK (C 0) - Me ₃ ) u (O) (o ' Aza cyPn) _r -CH -gassmaMeLeucínol c <0 J cMet) c eo-Gly-Asp ÝeyPaO- - - - '^ gáSagMeúeuc amino - N w -.- ..,. P R ₂ CHNKC (O) -Tbg-CH - (R) -CH- (CH ₂ C (O) CMe 1) -C (O) -Asp (cyPn) -NK- (R) -CH ( Ú) CMe ₃ , β ₂ CHNKC (O) -Tbg-CH ₂ - (R) -CH (CH ₂ C (O) CMe ₃ ) C (OJ-As? (Cyne) -NHCH-CMe, , Zt? R ₂ CNHC (O) -T 3 g-CH - (R) -CH (CH ₂ C (O) CMe ₃ ) C (C 1 -C 5? (Cylene) -NH. - (R) -CH ( Zt) CMe ₃ a Zc? R _? CNKC _(Oj-Tbg-CH? - (R) -CH (CH ₂ C (0) CMe) C (0) -A 5? (Cy? N) -NHCH ₂ CMe ,, weighing -nukle.os.idovýnr is analogen acyc lovir. A pharmaceutical composition for the treatment of acyclovir-resistant human herpes simplex virus, characterized in that it comprises, as its active ingredient, a peptide-denaturing formula. _in ____________ _ A-3-O-CH-CH / CH ₂ C (C) R 4 / C (O) -NKCH / C 2. ² (V) C3QH / C (O) - (U Ρ; ' ►where A is phenylacetyl, phenylpropionyl, (4-aminophenyl) propionyl, (4-fluorophenyl) propionyl, (4-hydroxyphenyl) propionyl, (4-methoxyphenyl) propionyl, 2- (phenylmethyl) -3-phenylpropionyl, 2 - / (4- fluorophenyl) methyl] -3- (4-fluorophenyl) propionyl, 2 - [(4-methoxyphenyl) methyl] -3- (4-methoxyphenyl) propionyl or benzylaminocarbonyl, B represents (N-Me) -Val or (N-Me) -Ile or forms together A and B are saturated alkylaminocarbonyl, butylaminocarbonyl, 1-methylethylaminocarbonyl, 1-methylpropylaminocarbonyl, 1-ethylpropylaminocarbonyl, 1,1-dimethylbutylaminocarbonyl, 1-ethylbutylaminocarbonyl, 1-propylbutylaminocarbonyl, 1-ethylpentylaminocarbonyl, 1-ethylpentylaminocarbonyl; outylaminocarbonyl, 2-ethylpentylaminocarbonyl, 1-methyl-1-propylbutylaminocarbonyl, 1-ethyl-1-propylbutylanubicarbonyl, 1,1-dipropylbutylaminocarbonyl, (1-propylcyclopentyl) aminocarbonyl or (1-propylcyclohexyl) aminocarbonyl, D stands for Val, Ile or Tbg, r! means 1-methylethyl, 1,1-dimethylethyl, 1-methylpropyl, 1,1-dimethylpropyl, 2,2-dimethylpropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-methylcyclopentyl, skudne NR ⁴ R ⁵ , where R ⁴ is hydrogen or lower 5 „.45 alkyl and R are lower alkyl or form R and R - together with the nitrogen atom to which they are attached a pyrrolidine, piperidine, morpholine or 4-methylpiperazine ring; R represents a hydrogen atom and q R is methyl, ethyl, 1-methylethyl, 1,1-dimethylethyl, propyl, 2-propenyl or benzyl, the carbon atom To which R and R groups are attached and in the configuration R, or R 3 represents a methyl group or a carbon atom to which the cyclobutyl, cyclopentyl or cyclohexyl are bonded and represents an NHR group wherein R is 2-methylpropyl; 1,2-Dimethylpropyl-1-Rf2T2 = trimethylpentyl, X1,22-1,16% of petroleum 1,1 (R) -ethyl 1- = 2,2 = dimethylpropyl. <A, - i - Me * weed i, l i-uinictiiy uutý Ϊ.Τ úxiíie ins ________ ^ -butyl., "L <R)., Exp.,. 2vbríme.thylbutyl · _r _ .. l. ( .R5 _..; .3.3-trimethyl .-, _ butyl, 2-ethylbutyl2,2-diethylbutyl. 2-ethyl-1 (R) -methylbutyl, 1,2-ethyl-2-methylbutyl, 1 (R) -ethyl-3,3-dimethylbutyl, 2,2-dimethylpentyl, cis- or trans-2-methylcyclohexyl 2,2-dimethylcyclohexyl or cyclohexylmethyl, or E is NHCH (R) -Z, wherein the carbon atom to which R is attached is S, R is 1,1-dimethylethyl, 1-methylpropyl, 2-methylpropyl, 2,2-dimethylpropyl or cyclohexylmethyl; and Z is CH-OH, C (O) OH, C (O) NH 88 or C (O) OR, wherein R is methyl, ethyl or propyl, or a pharmaceutically acceptable salt thereof, with a pharmaceutically acceptable carrier. 17. A pharmaceutical composition according to claim 16, characterized in that it comprises a combination of a pesticidal derivative of the general formula I according to claim 23, or a pharmaceutically acceptable salt thereof, and a nucleoside analogue having an anti-glidant effect, or a pharmaceutically acceptable salt thereof. ..... - - 18 ♦ armacauSičký proa.ťfe F ^ 'IFP <Jdrě ^- harokurí6 ^R 7 "y = 7 characterized in that as active component comprises a combination of a peptide derivative of one of the group vtarse« ι Γ HCS _CH.C P (Q) - (N-MeVal-Tb g-CH ₂ - (A) CS (CS? C (O) -CMe ₃₎ C (O) -As? (Cy? N) -gammaMeLeucínol , SS ₂ CHNHC (G) -7bg-CH ₂ - (a) -CS (CK ₂ C (G) CMe ₃ ) C (O) -Asp (cyne) -gammaMeucuino, P r, CSNHC (O) -Tb g-CS., - (R) -CS- (CH 2 C (O) CMe ₃ ) C (O) -As 2 (cyPn) -NK - ('a) -CK (& e) CM @ ₃ P r ^ CHNHC (O) -Tb g-CS ₂ - (R) -CS (CS ₂ C (O) CMe ₃ ) C (O) -Asp (cyne) -NKCS ₂ CMe ₃ , EC? * ₂ CNKC (O) -C ₂ -73 g - (A) CS (CS ₂ C (O) CMe) C (O) - As? (cyne) -NH- (R) -CS (Et) CMe ^a Ξ t? r ₂ CNKC (O) -Tb g-CS, - (R) —NSCS-CSe., 24 (CS-C (O) CH6) C (O) -AS2 (cyne) and acyclovir as a nucleoside analog. Represented by: ////%! * ¹ 0.8 '\ \ \ (Acyclovir) 0.6 0.4 AND\ 1/2 ti o tc o 0.2 0, 0.8 0 7 0.2 ‘0.4 7 0.6 fixed concentration of peptides from example TO EC _{eo of} peptides of Example 10