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WO1996040759A1 - Antiviral peptoid compounds - Google Patents

Antiviral peptoid compounds Download PDF

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Publication number
WO1996040759A1
WO1996040759A1 PCT/EP1996/002424 EP9602424W WO9640759A1 WO 1996040759 A1 WO1996040759 A1 WO 1996040759A1 EP 9602424 W EP9602424 W EP 9602424W WO 9640759 A1 WO9640759 A1 WO 9640759A1
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WO
WIPO (PCT)
Prior art keywords
lower alkyl
amino
phenyl
narg
bivalent radical
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Ceased
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PCT/EP1996/002424
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French (fr)
Inventor
Eduard Felder
François HAMY
Gerhard Heizmann
Thomas Klimkait
Janis Karlis Lazdins
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Novartis AG
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Ciba Geigy AG
Novartis AG
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Priority to EP96917496A priority Critical patent/EP0832110A1/en
Priority to AU60050/96A priority patent/AU6005096A/en
Publication of WO1996040759A1 publication Critical patent/WO1996040759A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/001—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof by chemical synthesis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to antiviral compounds comprising a peptoid structure, processes for the preparation of said compounds, pharmaceutical preparations comprising said compounds, the compounds for the use in the therapeutic (including prophylactic) or diagnostic treatment of the animal or especially human body, and the use of said compounds for the therapeutic or diagnostic treatment of the animal or especially human body or for the manufacture of pharmaceutical preparations.
  • HIV-1 a virus which is regarded as causative agent for the complex disease process leading to AIDS, encodes two regulatory proteins, Tat and Rev, which act through mechanisms the knowledge of which was unprecedented in the scientific community to determine both the quantity and quality of HIV-1 gene expression. These novel regulatory pathways are controlled at the level of protein-RNA interaction.
  • Two classes of HIV mRNAs can be distinguished. The first of these consists of a doubly spliced, 2 kb mRNA species that encodes the viral regulatory proteins, including Tat and Rev. The second class consists of the unspliced (9 kb) and incompletely spliced (4 kb) viral mRNAs that encode the virion structural proteins. Tat induces a marked increase in the steady-state level of viral mRNA.
  • RNA transcripts are fully spliced by the cellular RNA processing machinery prior to export to the cytoplasm.
  • Rev unspliced (Gag) or partially spliced (Env) viral mRNAs evade processing - instead they are exported directly to the cytoplasm.
  • Rev induces the efficient export of viral RNA species that are otherwise excluded from the cell cytoplasm.
  • Tat and Rev recognize regulatory elements on the viral mRNA. Tat function is mediated through a sequence termed TAR (for trans-activation response region) that comprises part of the 5 '-noncoding region of all HIV mRNAs.
  • Rev protein is highly sequence specific and requires recognition of an RNA target sequence, the Rev Responsive Element (RRE), a highly conserved region in the middle of the viral env gene.
  • RRE Rev Responsive Element
  • the RNA binding sites of both Tat and Rev map to protein areas which are highly arginine rich (see Calnan, B.J., et al., Science 252, 1167-1171 (1991) and Tiley, L.S., et al., Proc. Natl. Acad. Sci. USA 89, 758-62 (1992)).
  • Tat(l-86) The principle form of Tat (designated as Tat(l-86) herein) consists of 86 amino acids in known linear sequence (see Ratner et al., Nature 313, 277 (1985), which is inco ⁇ orated by reference herein).
  • Three domains in the protein have been shown to exist by structure/function analysis, including a proline-rich region spanning residues 1-18, a cysteine-rich region spanning residues 22-37, and a basic region of nine amino acid spanning residues 49-57 with the sequence 49 (L)-Arg-(L)-Lys-(L)-Lys-(L)-Arg-(L)-Arg-(L)-Gln-(L)-Arg-(L) -Arg-(L)-Arg 57 ,
  • basic region of the HIV Tat protein hereinafter.
  • the compounds of the present invention show very favourable and valuable pharmaceutical characteristics, especially with regard to the therapeutic and/or diagnostic treatment of retroviral infections, particularly AIDS.
  • the compounds of the invention comprising a peptoid structure provide synthetic structures with a comparatively low molecular weight which are effective in the treatment of HIV infection, especially HIV-1 infection, acting preferably on the basis of the mechanism described above.
  • the compounds of the invention comprising a peptoid structure represent a class of molecules with the ability to interfere with the Tat/TAR and Rev/RRE complex formations, inter alia with the following distinct advantageous characteristics:
  • the compounds of the invention act through a mechanism that provides them with an incomparable therapeutic potential to complement or replace existing, specific or less specific antiviral treatments, with particular value for the treatment against variants of HIV, especially such variants that have become resistant to other kinds of treatment.
  • An antiretroviral compound of the invention comprising a peptoid structure is preferably a peptoid compound of the formula I,
  • k 0 to 20
  • m 3 to 10
  • n 1 to 10
  • R ] represents hydrogen, acyl or an amino-substituent other than acyl
  • R 2 represents an OH group, a C-terminal protecting group or a primary, secondary or tertiary amino group
  • any A independently of the others being present represents a bivalent radical of an ⁇ -amino acid
  • any X independently of the others being present represents a bivalent radical of the partial formula II
  • each of R 3 , R 4 and R 5 independently of the others, represents hydrogen or a side chain of an ⁇ -amino acid other than glycine, or R 3 and R 4 together form an alkylene bridge and R 5 is hydrogen, or R 4 and R 5 together form an alkylene bridge and R 3 is hydrogen;
  • bivalent radicals X together form a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein, and
  • any B independently of the others being present represents a bivalent radical of an ⁇ -amino acid
  • the compounds of the present invention can exist as isomers or mixtures of isomers; for example, if one or more asymmetric carbon atoms are present, these carbon atoms can be in the (R)-, (S)- or (R,S)-configuration, independent of one another. It is thus possible to obtain isomeric mixtures, such as racemates or diastereomeric mixtures, or pure diastereomers or entantiomers, depending on the number of asymmetric carbon atoms and on whether isomers or isomeric mixtures are present. Preferred are pure isomers (enantiomers or diastereomers).
  • k is 0 to 20, preferably 0 to 12, most preferably 0 or 12; m is 3 to 10, preferably 4 to 6, most preferably 5; and n is 1 to 10, preferably 1 to 4.
  • lower defines a moiety with up to and including maximally 7, especially up to and including maximally 4, carbon atoms, said moiety being branched or straight-chained.
  • Lower alkyl for example, is methyl, ethyl, n-propyl, sec-propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl or n-heptyl.
  • Acyl has, for example, up to 25, preferably up to 19, carbon atoms and is especially the acyl group of a carboxylic acid or of a semiester of carbonic acid.
  • acyl groups of a carboxylic acid are unsubstituted or substituted alkanoyl having up to 19 carbon atoms, for example n-decanoyl, or preferably
  • lower alkanoyl such as formyl, acetyl, propionyl, butyryl or pivaloyl
  • substituted lower alkanoyl wherein preferably up to four, especially (except in the case of halogen which may be present up to three times as a substituent) up to two, substituents may be present, especially one substituent (except in the case of halogen which may be present up to three times as a substituent), the substituents being independently selected especially from
  • cycloalkyi with from 3 to 7 carbon atoms, especially in cycloalkyl-lower alkanoyl wherein lower alkanoyl is as defined above, for example cycloalkylcarbonyl, such as cyclopropyl-, cyclobutyl-, cyclopentyl- or cyclohexyl-carbonyl, or 2-cyclohexyl- or 2-cyclopentyl-acetyl,
  • aryl which has preferably from 6 to 14 ring carbon atoms, such as in phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, and may be unsubsti ⁇ tuted or mono- to tri-substituted especially by lower alkyl, for example methyl, ethyl or propyl, halo-lower alkyl, for example trifluoromethyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, for example methoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl- lower alkoxy or N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoyl amino, halogen, for example fluorine, chlorine or bromine, carb
  • heterocyclyl which is preferably a single or double ring system having from 3 to 10 ring atoms, is bonded via a carbon atom or, especially, via a nitrogen atom and contains up to 3 further hetero atoms selected from oxygen, nitrogen, sulfur, and sulfur linked to 1 or 2 oxygen atoms; which in addition may also be fused with 1 or 2 phenyl radicals or with 1 or 2 cycloalkyi radicals, cycloalkyi preferably having from 5 to 7 ring atoms; and which may be unsaturated or partially or fully saturated, for example thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl,
  • lower alkoxy especially in lower alkoxy-lower alkanoyl, for example lower alkoxy- acetyl or lower alkoxypropionyl, such as methoxyacetyl, ethoxyacetyl or 3-methoxypropionyl,
  • halo-lower alkanoyl containing up to 3 halogen atoms for example ⁇ -haloacetyl, such as ⁇ -fluoro-, ⁇ -chloro-, ⁇ -bromo-, ⁇ -iodo-, ⁇ , ⁇ , ⁇ -trifluoro- or ⁇ , ⁇ , ⁇ -trichloro-acetyl, or halopropionyl, such as ⁇ -chloro- or ⁇ - bromo-propionyl, and
  • aryl cycloalkyi
  • heterocyclyl are not restricted to the substitutents of lower alkanoyl, but are generally valid where these terms appear in the present specification, subject to the specific definitions of those moieties that, in each case, are characterized as being preferred.
  • a preferred acyl group of a semiester of carbonic acid has up to 19 carbon atoms and is selected preferably from the group comprising
  • lower alkoxycarbonyl for example methoxy-, ethoxy-, n-propoxy-, isopropoxy-, isobutoxy- or tert-lower alkoxy-carbonyl, or also or especially n-propoxycarbonyl, such as tert-butoxycarbonyl or isobutoxycarbonyl,
  • 2-halo-lower alkoxycarbonyl such as 2-chloro-, 2-bromo-, 2-iodo- or 2,2,2-trichloro- ethoxycarbonyl, aryl-lower alkoxycarbonyl, for example arylmethoxy-carbonyl, wherein aryl has from 6 to 14 carbon atoms and is, for example, phenyl, 1- or 2-naphthyl, fluorenyl, especially 9-fluorenyl, or phenyl mono- or poly-substituted by lower alkyl, for example methyl or tert-butyl, phenyl, hydroxy, lower alkoxy, for example methoxy, ethoxy or tert-butoxy, halogen, for example chlorine or bromine, and/or by nitro, for example phenyl-lower alkoxycarbonyl, such as benzyloxy carbonyl, 4-nitrobenzyloxycarbonyl, diphenylmethoxycarbon
  • heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is preferably a single or double ring system having from 3 to 10 ring atoms, is bonded via a carbon atom or, especially, via a nitrogen atom and contains up to 3 further hetero atoms selected from oxygen, nitrogen, sulfur, and sulfur linked to 1 or 2 oxygen atoms; which in addition may also be fused with 1 or 2 phenyl radicals or with 1 or 2 cycloalkyi radicals, cycloalkyi preferably having from 5 to 7 ring atoms; and which may be unsaturated or partially or fully saturated, for example thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, is
  • lower alkenyloxycarbonyl wherein preferably the lower alkenyl radical is bonded to the bonding oxygen atom via a saturated carbon atom, such as allyloxycarbonyl,
  • lower alkoxy-lower alkoxycarbonyl such as 2-methoxyethoxycarbonyl
  • (lower alkoxy-lower alkoxy )-lower alkoxycarbonyl such as 2-(2-methoxyethoxy)- ethoxycarbonyl
  • amino-substituent other than acyl is preferably an arylmethyl, etherified mercapto, 2-acyl-lower alk-1-enyl, a silyl group or an organic sulfonyl group.
  • each aryl radical has preferably from 6 to 14 ring carbon atoms, such as in phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, and may be unsubstituted or mono- to tri-substituted especially by lower alkyl, for example methyl, ethyl or propyl, halo-lower alkyl, for example trifluoromethyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, for example methoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy or N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkyl ⁇ amino,
  • acyl is, preferably, the corresponding radical of a lower alkanoic acid, of a benzoic acid that is unsubstituted or substituted, for example, by lower alkyl, such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen, such as chlorine, and/or by nitro, or especially of a carbonic acid semiester, such as a carbonic acid lower alkyl semiester.
  • lower alkyl such as methyl or tert-butyl
  • lower alkoxy such as methoxy
  • halogen such as chlorine
  • nitro or especially of a carbonic acid semiester, such as a carbonic acid lower alkyl semiester.
  • Corresponding groups are especially 1-lower alkanoyl-lower alk-l-en-2-yl, for example 1-lower alkanoylprop-l-en-2-yl, such as 1-acetyl- prop-l-en-2-yl, or lower alkoxycarbonyl-lower alk-l-en-2-yl, for example lower alkoxycarbonylprop-l-en-2-yl, such as l-ethoxycarbonylprop-l-en-2-yl.
  • a siiyl group is, for example, a tri-lower alkylsilyl group, for example trimethylsilyl or tert-butyl-dimethylsilyl.
  • a C-terminal protecting group is preferably an esterifying group, thus leading to an esterified C-terminal carboxy group. More preferred is a lower alkoxy group that is preferably branched in the 1 -position of the lower alkoxy group or substituted in the 1- or 2-position of the lower alkoxy group by suitable substituents.
  • a lower alkoxy group that is branched in the 1 -position of the lower alkoxy group is, for example, tert-lower alkoxy, for example tert-butoxy.
  • a lower alkoxy group that is substituted in the 1- or 2-position of the lower alkoxy group by suitable substituents is, for example, arylmethoxy having one or two aryl radicals, wherein aryl is preferably phenyl that is unsubstituted or mono-, di- or tri-substituted, for example, by lower alkyl, for example tert-lower alkyl, such as tert-butyl, lower alkoxy, for example methoxy, hydroxy, halogen, for example chlorine, and/or by nitro, for example benzyloxy, benzyloxy substituted by the mentioned substituents, for example 4-nitro- benzyloxy or 4-methoxybenzyloxy, diphenylmethoxy or diphenylmethoxy substituted by the mentioned substituents, for example di(4-methoxyphenyl)methoxy; 1-lower alkoxy- lower alkoxy, for example methoxymethoxy, 1 -methoxy
  • a C-terminal protecting group can furthermore be an organic silyloxy group.
  • An organic silyloxy group is, for example, a tri-lower alkylsilyloxy group, for example trimethylsilyloxy.
  • the silicon atom of the silyloxy group can also be substituted by two lower alkyl groups, for example methyl groups.
  • a C-terminal protecting group is preferably tert-lower alkoxy, for example tert-butyloxy, benzyloxy, 4-nitrobenzyloxy, 9-fluorenylmethoxy or diphenylmethoxy.
  • a primary, secondary or tertiary amino group is preferably a free amino group, a mono- or disubstituted amino group the substituents of which are preferably selected from the group comprising lower alkyl, e.g. methyl or ethyl, aryl-lower alkyl, such as phenyl-lower alkyl, e.g. benzyl, or heterocyclyl-lower alkyl, such as pyrrolidinyl-lower alkyl, e.g. 2-(l-pyrrolidinyl)-ethyl, pyridyl-lower alkyl, e.g.
  • a disubstituted amino group may also be N-containing heterocyclyl bonded via its nitrogen atom, such as e.g. 1-pyrrolidinyl or 4-mo ⁇ holinyl.
  • a bivalent radical of an ⁇ -amino acid is preferably bonded N-terminally by way of its ⁇ -amino group and C-terminally by way of its carboxy group and is preferably selected from the group comprising a bivalent radical of a natural ⁇ -amino acid having the L-conf ⁇ guration, such as those normally occurring in proteins, or an epimer of such an amino acid, that is to say having the unnatural D-configuration, or a D,L-isomeric mixture thereof; or a homologue of such an amino acid, for example wherein the amino acid side chain has been shortened by one or two methylene groups or lengthened to up to 10 carbon atoms, such as an ⁇ -amino alkanoic acid with 5 up to and including 10 carbon atoms in a linear chain, a substituted aromatic ( ⁇ -aryl or ⁇ -aryl lower alkyl) amino acid wherein the aryl radical has from 6 to 14 carbon atoms, for example a substituted phenylalanine or phen
  • the bivalent radical bonded via its ⁇ -amino and its ⁇ -carbonyl group, of an amino acid selected from glycine (H-Gly-OH), alanine (H-Ala-OH), valine (H-Val-OH), norvaline ( ⁇ -aminovaleric acid), leucine (H-Leu-OH), isoleucine (H-Ile-OH), norleucine ( ⁇ -aminohexanoic acid, H-Nle-OH), ⁇ -amino-n-decanoic acid, serine (H-Ser-OH), homoserine ( ⁇ -amino- ⁇ -hydroxybutyric acid), threonine (H-Thr-OH), methionine (H-Met-OH), cysteine (H-Cys-OH), proline (H-Pro-OH), trans-3- and trans-4-hydroxyproline, phenylalanine (H-Phe-OH), tyrosine (H-Tyr-OH),
  • each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form, preferably in the L- or in the D-form.
  • a side chain of an ⁇ -amino acid other than glycine and proline is the group other than the amino and the carboxy group bound to the ⁇ -carbon of the respective amino acid, that is a group R 6 that is bound to an amino acid structure of the formula III
  • R 6 is a side chain selected so as to give an ⁇ -amino acid (except for glycine where the residue corresponding to R 6 would be H and except for proline), more specifically one of the ⁇ -amino acids as defined above; more preferably, R 5 is a residue from the group comprising lower alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or sec-butyl, or substituted lower alkyl, such as aryl-lower alkyl wherein the aryl is preferably phenyl that may be mono- or poly-substituted by lower alkyl, for example methyl, lower alkoxy, for example methoxy, lower alkanoyloxy, for example acetoxy, amino, lower alkylamino, for example methylamino, di-lower alkylamino, for example dimethylamino, lower alkanoylamino, for example ace
  • alkyl preferably means lower alkyl, especially C r C 4 -alkyl, such a methyl or ethyl.
  • the term "comprising a peptoid structure” means that the compounds of formula I are oligoamidic compounds with at least one residue X of formula II in formula I wherein R 3 is not hydrogen, but a side chain of an amino acid other than glycine and proline.
  • TAR-binding perferably means binding as measured in the Tat-TAR gel-shift assay described below.
  • m is 4 to 9, preferably 4 to 6; and the basic side chains R 3 , R 4 and/or R 5 mentioned in the last paragraph above are preferred, with the exception of the X at position 4 of the residue -(X) m - (counted from the N-terminus, that is the underlined moiety in the following representation of the residue -(X) m -, namely -X-X-X-X- ...) where any of the side chains mentioned above, especially in the last paragraph, is possible; and more preferably
  • each X is a bivalent radical of the partial formula II
  • R 3 represents a side chain selected from the group comprising lower alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or sec-butyl, or substituted lower alkyl, such as aryl-lower alkyl wherein the aryl is preferably phenyl that may be mono- or poly-substituted by lower alkyl, for example methyl, lower alkoxy, for example methoxy, lower alkanoyloxy, for example acetoxy, amino, lower alkylamino, for example methylamino, di-lower alkylamino, for example dimethylamino, lower alkanoylamino, for example acetylamino or pivaloylamino, lower alkoxycarbonylamino, for example tert-butoxycarbonylamino, arylmethoxycarbonylamino wherein aryl preferably has from 6 to 14 carbon
  • each of R 4 and R 5 represents hydrogen.
  • a bivalent radical of an ⁇ -amino acid is preferably a bivalent radical of an amino acid selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present preferably in the L-form (where an asymmetric ⁇ -carbon atom is present).
  • a bivalent radical of an ⁇ -amino acid is preferably a bivalent radical of an amino acid selected from lysine, arginine and proline, each of which is present preferably in the D-form.
  • a and B are selected so as not to interfere with the binding of the respective compound of formula I to the TAR target.
  • Most preferred are sequences that are analogous (with small deviations, such as conservative substitutions of amino acids, e.g. up to three such substitutions ) or identical to those in the corresponding Tat (1-86).
  • Salts of compounds of formula I are especially acid addition salts, salts with bases or, where several salt-forming groups are present, can also be mixed salts or internal salts.
  • Salts are especially pharmaceutically acceptable salts of compounds of formula I.
  • Such salts are formed, for example, from compounds of formula I having an acid group, for example a carboxy group, a sulfo group, or a phosphoryl group substituted by one or two hydroxy groups, and are, for example, salts thereof with suitable bases, such as non-toxic metal salts derived from metals of groups Ia, lb, Ila and lib of the Periodic Table of the Elements, especially suitable alkali metal salts, for example lithium, sodium or potassium salts, or alkaline earth metal salts, for example magnesium or calcium salts, also zinc salts or ammonium salts, as well as salts formed with organic amines, such as unsubstituted or hydroxy-substituted mono-, di- or tri-alkylamines, especially mono-, di- or tri-lower alkylamines, or with quaternary ammonium compounds, for example with N-methyl-N-ethylamine, diethylamine, triethylamine, mono-, bis-
  • the compounds of formula I having a basic group, for example an amino group can form acid addition salts, for example with inorganic acids, for example hydrohalic acids, such as hydrochloric acid, sulfuric acid or phosphoric acid, or with organic carboxylic, sulfonic, sulfo or phospho acids or N-substituted sulfamic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methyl- maleic acid, fumaric acid, malic acid, tartaric acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosali- cylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, as well as with amino acids, for example the ⁇ -amino
  • the compounds of the invention have useful, in particular pharmacologically useful, properties.
  • the compounds of formula I are able to inhibit the propagation of HIV, especially HIV-1, in infected human lymphocytes and show a particularly potent, specific inhibition on the binding of the Tat protein to TAR, mainly by binding to TAR. They thus represent a totally new class of inhibitors and therapeutics.
  • the in vitro inhibition of the interaction between Tat and TAR can be shown by a competition Tat-TAR gel-shift assay.
  • the sequence of recombinant Tat can be found in Churcher et al., J. Mol. Biol. 230, 90-110 (1993) to the RNA (synthetic TAR duplex, Genset, Paris, France; the sequence can be found in Hamy et al., J. Mol. Biol. 230, 111-123 (1993)), the overall size and the charge/weight ratio of the formed duplex are changed, so that electrophoretic migration through a native polyacrylamide gel is affected.
  • RNA and complexes can be discriminated based on their relative positions in the gel (Hamy et al., J. Mol. Biol. 230, 111-123 (1993)). If the binding reaction with a substance able to prevent the protein binding to the radiolabelled RNA, this competition for binding can be visualized on the autoradiography as a decreased intensity of the retarded band.
  • compounds of the formula I are tested as follows:
  • the uninco ⁇ orated [ ⁇ -32P]ATP is removed by chromatography through a sephadex NAP- 10 column (Pharmacia, Uppsala, Sweden) equilibrated with water.
  • the labelled 14-mer is annealed to 1.5 equivalents of unlabelled 17-mer by heating to 90 °C for 3 min, followed by slow cooling down to 0 °C.
  • the binding reaction for protein and RNA which takes place in a volume of 25 ⁇ l contains approximately 10,000 cpm of the labelled duplex TAR-RNA and 20 nM recombinant Tat protein in TK buffer (Tris-HCl 20 mM pH 8.0, KCl 50 mM) with 10 mM DTT, 0.1 % Triton X-100 ((Alkylphenylpolyethylenglykol, Rohm & Haas, Darmstadt, Germany) in the absence or presence of varying concentrations of inhibitor.
  • TK buffer Tris-HCl 20 mM pH 8.0, KCl 50 mM
  • Triton X-100 (Alkylphenylpolyethylenglykol, Rohm & Haas, Darmstadt, Germany) in the absence or presence of varying concentrations of inhibitor.
  • the autoradiographies are quantified by Phosphorimager (Molecular Dynamics/Bucher, Basle, Switzerland).
  • a CD50 value is determined as the concentration of a compound of the formula I giving a 50 % decrease in the intensity of the retarded band (Tat-TAR complex).
  • the CD50-values that are obtained are preferably in the range of from 1 x 10" 9 to 1 x 10" 6 M. It is possible to show that similar binding affinity can be found when wild-type unlabelled TAR-RNA is used as competitor, thus suggesting that the compounds of the present invention have affinities comparable to that of the high molecular weight full-length Tat protein in vitro.
  • the compounds of the present invention can also be shown to inhibit Rev/RRE interaction in a competition Rev-RRE gel-shift assay (see Kjems, J., et al., EMBO J. 11, 1119-1129 (1992) and Tan, R., et al., Cell 73, 1031-40 (1993).
  • PBLs Peripheral Blood Mononuclear Lymphocytes
  • Cells (1 x 10 6 /ml) are cultured for 2 days in RPMI-1640 (Gibco), supplemented with 10 % heat-inactivated fetal calf serum (Gibco), 50 ⁇ g ml streptomycin, 50 U/ml penicillin (Amimed), 2 nM glutamine and 10 mM hepes buffer (Gibco).
  • Stimulated lymphocytes are obtained by culturing in the presence of PHA (0.25 ⁇ g/ml; Wellcome diagnostics, Templehill, Dartford, England). PHA-lymphocyte stimulation is confirmed by the increase in cell size (Scattergram, FACS analysis).
  • RT determination is possible as follows: The RT activity is determined in 50 mM of tris ( ⁇ , ⁇ , ⁇ -tris(hydroxymethyl)methylamine, ultra pure, Merck, Federal Republic of Germany) pH 7.8; 75 mM of KCl, 2 mM of dithiothreitol, 5 mM of MgCl2; 0.05 % Nonidet P-40 (detergent; Sigma, Switzerland); 50 ⁇ g/ml of polyadenylic acid (Pharmacia, Sweden); 1.6 ⁇ g/ml of dT(12-18) (Sigma, Switzerland). The mixture is filtered through an Acrodisc filter (0.45 ⁇ : Gellman Science Inc, Ann Arbor) and stored at -20°C.
  • Acrodisc filter (0.45 ⁇ : Gellman Science Inc, Ann Arbor
  • the RT activity is a measure of the reproduction of HIV-1.
  • the compounds of formula I according to the invention inhibit virus reproduction when administered in the micromolar range, for example during 18 days after infection practically no increase in RT activity can be determined in the presence of preferably 5 to 50 ⁇ M concentrations of an inhibitor of the present invention (for example 0 to 100 counts per minute), while in the control high increase of RT activity can be found (for example more than 2000 counts per minute on day 18).
  • the compounds of the present invention can thus be used in the treatment of retroviral infections in warm-blooded animals, especially HIV, such as HTV-l, infections, and more specifically for the treatment of AIDS in humans.
  • treatment of infected cells, e.g. lymphocytes, outside the body is possible in order to reintroduce healthy cells by transplantantion or injection, for example in order to improve the lymphocyte titer in patients with advanced AIDS.
  • the compounds of the present invention can also be used in the treatment of commercially valuable cell, such as lymphocyte, cultures against retroviral infections, especially against HIV, such as HTV-l, infections.
  • k is 0 to 20, preferably 0 to 12, most preferably 0 or 12, m is 3 to 10, preferably 4 to 6, most preferably 5, n is 1 to 10, preferably 1 to 4;
  • substituted lower alkanoyl wherein the substituents are independently selected from - cycloalkyi with from 3 to 7 carbon atoms; - aryl selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, each of which is unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, nap
  • heterocyclyl selected from thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, chromanyl, cyclohexa[b]pyrrolyl, cyclohexa- [bjpyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, mo ⁇ holinyl, thiomo ⁇ holinyl, S,S-dioxo-thiomo ⁇ holinyl
  • aryl-lower alkoxycarbonyl wherein aryl is phenyl, 1- or 2-naphthyl, fluorenyl or phenyl mono- or poly-substituted by lower alkyl, phenyl, hydroxy, lower alkoxy, halogen or nitro,
  • heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, - pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, iso- quinolyl, 3,1-benzofuranyl, cyclohexa[b]pynolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, mo ⁇ holinyl, thiomo ⁇ holinyl, S,S- dio
  • aryl-lower alkyl with up to three aryl groups, wherein each aryl is selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl which are unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di- lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl
  • arylthio or aryl-lower alkylthio, wherein aryl is phenyl that is unsubstituted or substituted by lower alkyl, lower alkoxy, halogen or nitro,
  • arylsulfonyl wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups,
  • arylmethoxy having one or two aryl radicals, wherein aryl is phenyl that is unsubstituted or mono-, di- or tri-substituted by lower alkyl, lower alkoxy, hydroxy, halogen or nitro,
  • a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyrrolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, mo ⁇ holinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyrrolidinyl an 4-mo ⁇ holinyl;
  • any A being present and any B being present are independently selected from the group comprising a bivalent radical, bonded via its ⁇ -amino and its ⁇ -carbonyl group, of an amino acid selected from glycine alanine, valine, norvaline, leucine, isoleucine, norleucine, ⁇ -amino-n-decanoic acid, serine, homoserine, threonine, methionine, cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine, tyrosine, 4-amino- phenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenyl- alanine, ⁇ -phenylserine,phenylglycine, ⁇ -naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan, indoline-2-carboxylic
  • each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form, preferably in the L-form (more preferred in the case of the radical A) or in the D-form (more preferred in the case of the radical B);
  • the moiety A being a bivalent radical of an ⁇ -amino acid selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present preferably in the L-form (where an asymmetric ⁇ -carbon atom is present), the moiety B being a bivalent radical of an ⁇ -amino acid selected from lysine, arginine and proline, each of which is present preferably in the D-form;
  • each X being present is a bivalent radical of the partial formula II,
  • R 3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
  • - amino-lower alkyl such as 4-aminobutyl or 6-aminohexyl
  • N-lower alkylamino- such as N-methylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, N,N-di(phenyl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or guanidino-lower alkyl, such as 3-guanidinopropyl (lower alkyl preferably being methyl, ethyl or propyl),
  • - pyridyl-lower alkyl for example (2-pyridyl)-lower alkyl, such as 2-(2- ⁇ yridyl)ethyl,
  • - pyrrolidinyl-lower alkyl for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pynolidinyl)ethyl,
  • - mo ⁇ holinyl-lower alkyl for example (4-mo ⁇ holinyl)-lower alkyl, such as 2-(4-mo ⁇ holinyl)ethyl,
  • residue X which is in position 4 of the bivalent residue -(X) m - when counted from the N-terminus may in addition be a side chain selected from
  • aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
  • - furyl-lower alkyl for example (2-furyl)-lower alkyl, such as (2-furyl)methyl, and
  • indolyl-lower alkyl for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl;
  • R 4 and R 5 each represent hydrogen
  • k is 0 to 12, more preferably 0 or 12, most preferably 0, m is 4 to 6, most preferably 5, n is 1 to 4;
  • R 2 is amino, or further
  • a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyrrolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, mo ⁇ holinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyrrolidinyl an 4-mo ⁇ holinyl;
  • any A being present is a bivalent radical of an ⁇ -amino acid bound at its N-terminus via its ⁇ -amino group and at its C-terminus via its ⁇ -carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or preferably in the L-form (where an asymmetric ⁇ -carbon atom is present);
  • any B being present is a bivalent radical of an ⁇ -amino acid bound at its N-terminus via its ⁇ -amino group and at its C-terminus via its ⁇ -carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
  • R 3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
  • - amino-lower alkyl such as 4-aminobutyl or 6-aminohexyl
  • N-lower alkylamino- such as N-methylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, N,N-di(phenyl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or guanidino-lower alkyl, such as 3-guanidinopropyl (lower alkyl preferably being methyl, ethyl or propyl),
  • - pyridyl-lower alkyl for example (2-pyridyl)-lower alkyl, such as 2-(2-pyridyl)ethyl,
  • - pyrrolidinyl-lower alkyl for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pyrrolidinyl)ethyl, and
  • - mo ⁇ holinyl-lower alkyl for example (4-mo ⁇ holinyl)-lower alkyl, such as 2-(4-mo ⁇ holinyl)ethyl, or, in the case of the residue X which is in position 4 of the bivalent residue -(X) m - when counted from the N-terminus (that is the underlined moiety in the following representation of the bivalent radical -(X) m -, namely -X-X-X-X- 7), may in addition be a side chain selected from
  • aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
  • - furyl-lower alkyl for example (2-furyl)-lower alkyl, such as (2-furyl)methyl, and
  • indolyl-lower alkyl for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl;
  • - amino-lower alkyl such as 4-aminobutyl or 6-aminohexyl, or
  • aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, aryl ⁇ methoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro; phenyl-lower alkyl, especially benzyl, being most preferred in that position;
  • R 4 and R 5 each represent hydrogen
  • k is 0 or 12, most preferably 0, m is 4 to 6, most preferably 5, n is 1 to 4;
  • R is ammo
  • any A being present is a bivalent radical of an ⁇ -amino acid bound at its N-terminus via its ⁇ -amino group and at its C-terminus via its ⁇ -carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or preferably in the L-form (where an asymmetric ⁇ -carbon atom is present);
  • any B being present is a bivalent radical of an ⁇ -amino acid bound at its N-terminus via its ⁇ -amino group and at its C-terminus via its ⁇ -carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
  • R 3 represents a side chain selected from the group comprising
  • - amino-lower alkyl such as 4-aminobutyl or 6-aminohexyl
  • guanidino-lower alkyl such as 3-guanidinopropyl
  • residue X which is in position 4 of the bivalent residue -(X) m - when counted from the N-terminus that is the underlined moiety in the following representation of the bivalent radical -(X) m -, namely -X-X-X-X- ...), may in addition be a phenyl-lower alkyl side chain, such as benzyl;
  • R and R 5 each represent hydrogen
  • the compounds of the present invention can be synthesized according to known procedures, especially by a process comprising
  • R 3 is a residue from the group comprising lower alkyl or substituted lower alkyl (preferably as defined above for R 6 in formula III), while R 2 , R 4 , R 5 , A, B, X, m and n have the meanings given for compounds of the formula I,
  • R 2 ' has the same meaning as R 2 in compounds of formula I or is a resin for solid phase synthesis and R 4 , R 5 , X, B, m and n have the meanings given for compounds of formula I, under nucleophilic substitution, in the mentioned starting materials free functional groups with the exception of those that participate in the reaction if required being present in protected form; and removing any protecting groups and cleaving from any resin for solid phase synthesis being present;
  • R j , R 2 , R 3 , R 4 , R 5 , A, B, X, k, m and n have the meanings given for compounds of the formula I, if not mentioned otherwise.
  • the compounds of the present invention preferably can be readily prepared according to well-established, standard liquid or. preferably, solid-phase peptide synthesis methods, general descriptions of which are broadly availabe, for example in J.M. Stewart and J.D. Young, Solid Phase Peptide Synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984), in M. Bodanzsky and A. Bodanzsky, The Practice of Peptide Synthesis, Springer Verlag, New York (1984); and Applied Biosystems 430A Users Manual, ABI Inc., Foster City, California, but may also be prepared in solution or by a combination of solid-phase and solution chemistry.
  • a fragment with a free carboxy group can be an amino acid (if required, in suitably protected form) or a di-, tri- or other oligopeptide (the term "peptide” here also comprising peptoid structures) or also, e.g. in the case of the synthesis of derivatives of formula I with acylated terminal amino group, the acylating carbonic acid, especially acetic acid.
  • a fragment that has an amino group with at least one free hydrogen atom can also be a single amino acid, a di-, tri- or oligopeptide or, in the case of preparation of peptamides, ammonia or mono- or disubstituted ammonia.
  • Reactive derivatives of carbonic acids are preferably reactive esters or reactive anhydrides, or reactive cyclic amides. Reactive carbonic acid derivatives can also be formed in situ.
  • a reactive derivative of an "amino group with at least one free hydrogen” is preferably derivatized by the reaction with a phosphite, such as diethyl-chlorophosphite, 1,2-phenylene-chlorophosphite, ethyl-dichlorophosphite, ethylene-chlorophosphite or tetraethyl-pyrophosphite; or is present in the form of a carbamic acid chloride wherein the amino group participating in the reaction is subtituted by halocarbonyl, such as chlorocarbonyl.
  • a phosphite such as diethyl-chlorophosphite, 1,2-phenylene-chlorophosphite, ethyl-dichlorophosphite, ethylene-chlorophosphite or tetraethyl-pyrophosphite
  • a phosphite
  • the reactions normally run in the presence of a condensing agent or, when activating the carboxylic acids in the form of anhydrides, of an agent that binds the carboxylic acid formed. In some cases it is also possible to add chaotropic agents such as LiF in NB-methylpynolidone.
  • the reactions are especially carried out in a temperature range from -30 to +150 °C, preferably from +10 to +70 °C, and, most preferably, from +20 to +50 °C, if appropriate, in an inert gas atmosphere, e.g. under nitrogen or argon.
  • unreacted amino groups can be acylated after a reaction cycle, e.g. by acetylation of unreacted amino groups with 10 ml acetic anhydride/pyridine/DMA (1:1:8), thus facilitating later purification of the final product.
  • a suitably protected amino acid an ⁇ -amino acid or (for the introduction of peptoid structures) an amino acid of formula lib
  • R 3 , R 4 and R 5 have the meaning given under formula II above, R 3 not being hydrogen] as a ligand is attached via its carboxyl group (-COOH) to a derivatized, insoluble polymeric support, e.g. a cross-linked styrene or polyamide resin, such as a 4-(2',4'-dimethoxyphenyl-[hydroxy- or amino-]methyl)-phenyoxymethyl-polystyrene resin by a condensation reaction.
  • "Suitably protected” refers to the presence of protecting groups on the ⁇ -amino group ( ⁇ -NH 2 or ⁇ -NHR 3 ) and any side-chain functional group (if present) of the amino acid. Di-, tri- or other oligopeptides can be used instead of the amino acids as building blocks (fragments).
  • Synthesis proceeds in a stepwise, cyclical fashion by successively removing the ⁇ -NH or ⁇ -NHR 3 protecting group and then coupling an activated fragment (e.g. an amino acid, di-, tri- or oligopeptide) to the deprotected ⁇ -NH 2 or ⁇ -NHR 3 .
  • an activated fragment e.g. an amino acid, di-, tri- or oligopeptide
  • activation of the ⁇ -COOH group of the amino acid to be attached by the condensation reaction is effected (i) directly with a carbodiimide, e.g.
  • DCC dicyclohexylcarbodiimide
  • a carbonyl compound such as carbonyldiimidazole
  • 1,2-ox- azolium compounds such as 2-ethyl-5-phenyl-l,2-oxazolium-3 '-sulfonate and 2-tert- butyl-5-methylisoxazolium perchlorate
  • acylamino compounds such as 2-ethoxy- l-ethoxycarbonyl-l,2-dihydroquinoline
  • an uronium compound such as 2-(lH-benzo- triazol-l-yl)-l,l,3,3-tetramethyluronium tetrafluoroborate (TBTU
  • an "active ester” e.g. a hydroxybenzotriazole (HOBT), pentafluorophenyl, 4-nitrophenyl or N-hydroxysuccinimide ester.
  • Useful acid binding agents that can be employed in the condensation reactions are, for example, alkaline metals, carbonates or bicarbonates, such as sodium or potassium carbonate or bicarbonate (if appropriate, together with a sulfate), or organic bases such as sterically hindered organic nitrogen bases, for example tri-lower alkylamines, such as N,N-diisopropyl-N-ethylamine,
  • Reactive groups in the monomers of ligands or in the resin-bound or free intermediates resulting from one or more coupling steps can be protected by third groups as protecting groups that are customarily used in peptide synthesis.
  • third groups protecting groups that are customarily used in peptide synthesis. Examples of protecting groups, their introduction and their removal are, for example, described in standard works such as "Protective groups in Organic Chemistry", Plenum Press, London, New York 1973; “Methoden der organischen Chemie”, Houben-Weyl, 4. edition, Vol. 15/1, Georg-Thieme Verlag, Stuttgart 1974; Th. W.
  • protecting groups comprises also resins used for solid phase synthesis, preferably those specifically mentioned above and below.
  • hydroxy protecting groups are acyl radicals, such as tert-lower alkoxycarbonyl radicals, for example tert-butoxycarbonyl, etherifying groups, such as tert-lower alkyl groups, for example t-butyl, or silyl- or tin radicals, such as tert-butyl-dimethylsilyl or the tri-n-butyltin radical.
  • acyl radicals such as tert-lower alkoxycarbonyl radicals, for example tert-butoxycarbonyl
  • etherifying groups such as tert-lower alkyl groups, for example t-butyl
  • silyl- or tin radicals such as tert-butyl-dimethylsilyl or the tri-n-butyltin radical.
  • Carboxy groups can be protected by groups as defined above for the C-terminal protecting groups R 2 , preferably by esterifying groups selected from those of the tert-butyl type, from benzyl, from trimethylsilylethyl and from 2-triphenylsilyl groups .
  • Amino or guanidino groups can be protected by removable acyl groups or by arylmethyl, etherified mercapto, 2-acyl-lower alk-1-enyl, a silyl group or an organic sulfonyl group (preferably as defined above for an "amino substituent other than acyl" R j ) or tin amino protecting groups; tert-butoxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl, 4-nitro- benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2-bromobenzyloxycarbonyl (especially the tyrosine OH group), diphenylmethoxycarbonyl, nitrophenylsulfenyl, 2,2,2-trichloro- ethoxycarbonyl, 2,2,5,7,8-pentamefhylchroman-6-sulfonyl (PMC), 2,2,4,6,7-pentamethyl- dihydrobenz
  • Imino groups (e.g. in imidazole) can be protected by 2,4-dinitrophenyl or p-toluene- sulfonyl, or (e.g. in indole) by formyl.
  • Mercapto groups can be protected, e.g., by acetamidomethyl, by trityl or by p-methylbenzyl.
  • protective groups are usually removed after the complete synthesis of the resin-bound molecule by conventional methods of peptide chemistry, conveniently by treatment with 95 % trifluoroacetic acid.
  • strong nucleophiles such as 1,2-ethanedithiol, may be additionally added to capture the generated compounds resulting from the protecting groups.
  • Groups of the ⁇ -elimination type are typically protective groups of the fluorenylmethyl type.
  • the two prefe ⁇ ed methods of solid phase peptide synthesis are the Boc and the Fmoc methods, which are named with reference to their use of the tert-butoxycarbonyl (Boc) or 9-fluorenylmethyloxycarbonyl (Fmoc) group, respectively, to protect the ⁇ -NH 2 or ⁇ -NHR 3 of the amino acid residue to be coupled.
  • TFA trifluoroacetic acid
  • Prefe ⁇ ed third groups as protecting groups (for functional groups in side chains) are relatively stable in weak acid, e.g. TFA. Most can be cleaved by strong acids such as hydrofluric acid (HF) or trifluoromethanesulfonic acid.
  • HF hydrofluric acid
  • a small number of side chain groups e.g. 2,4-dinitrophenyl protected imino in the histidyl side chain, may require a separate deprotection step, e.g. treatment with thiophenol or ammonolysis.
  • the product is typically cleaved from the resin and simultaneously deprotected by HF treatment at low temperature (e.g. around 0 °C).
  • the Fmoc-group can be cleaved off preferably in the presence of a mild nitrogen base, preferably piperidine, in an inert solvent, preferably dimethyl acetamide, thereby allowing the use of side-chain protecting groups which are labile to milder treatment, e.g. TFA.
  • a mild nitrogen base preferably piperidine
  • an inert solvent preferably dimethyl acetamide
  • An acid labile ether resin such as HMP-resin (p-hydroxymethylphenoxymethyl poly ⁇ styrene), 4-(2',4'-dimethoxyphenyl-hydroxymethyl)-phenoxymethyl-polystyrene or preferably a resin with a benzyloxy- or alkyloxy linker (see Wang, J. Amer. Chem. Soc. 95, 1328 (1973); or, for the synthesis of compounds with a C-terminal amino group R 2 (in amide bond) which are prefe ⁇ ed, 4-(2',4'-dimethoxyphenyl-aminomethyl)-phenoxy- methyl-polystyrene (Rink et al., Tetrahedr. Lett. 28(33), 3787-90 (1987) is used as the solid support, permitting simultaneous cleavage/deprotection in TFA.
  • HMP-resin p-hydroxymethylphenoxymethyl poly ⁇ styrene
  • Z is preferably a nucleofugal group, preferably aryl- sulfonyloxy, such as toluenesulfonyloxy, lower alkanesulfonyloxy, such as methane- sulfonyloxy, or especially halogen, such as chlorine, bromine or iodine, more especially chlorine or iodine and most especially bromine.
  • aryl- sulfonyloxy such as toluenesulfonyloxy
  • lower alkanesulfonyloxy such as methane- sulfonyloxy
  • halogen such as chlorine, bromine or iodine, more especially chlorine or iodine and most especially bromine.
  • aprotic solvents such as ketones, for example a di-lower alkyl ketone, such as acetone, nitriles, for example a lower alkylnitrile, such as acetonitrile, carboxylic acid amides, for example a di-lower alkyl-lower alkanoylamide, such as dimethylformamide or dimethylacetamide, di-lower alkyl sulfoxides, such as dimethyl sulfoxide, hexamethyl- phosphoric acid triamide or ethers, such as di-lower alkyl ethers, for example diethyl ether, or cyclic ethers, such as tetrahydrofuran or dioxane, or also in protic solvents, such as alcohols, especially lower alkanols, for example methanol or ethanol, or mixtures of two or more of the mentioned
  • aprotic solvents such as ketones, for example a di-lower alkyl ket
  • the protecting groups, their introduction, their removal, the resins used for solid phase synthesis and methods of cleavage from them are preferably analogous to those described under process b).
  • Salts of compounds of formula I having at least one salt-forming group may be prepared in a manner known er se.
  • salts of compounds of formula I having acid groups may be formed, for example, by treating the compounds with metal compounds, such as alkali metal salts of suitable organic carboxylic acids, e.g. the sodium salt of 2-ethyl- hexanoic acid, with organic alkali metal or alkaline earth metal compounds, such as the co ⁇ esponding hydroxides, carbonates or hydrogen carbonates, such as sodium or potassium hydroxide, carbonate or hydrogen carbonate, with co ⁇ esponding calcium compounds or with ammonia or a suitable organic amine, stoichiometric amounts or only a small excess of the salt-forming agent preferably being used.
  • metal compounds such as alkali metal salts of suitable organic carboxylic acids, e.g. the sodium salt of 2-ethyl- hexanoic acid
  • organic alkali metal or alkaline earth metal compounds such as the co ⁇ espond
  • Acid addition salts of compounds of formula I are obtained in customary manner, e.g. by treating the compounds with an acid or a suitable anion exchange reagent.
  • Internal salts of compounds of formula I containing acid and basic salt-forming groups, e.g. a free carboxy group and a free amino group, may be formed, e.g. by the neutralisation of salts, such as acid addition salts, to the isoelectric point, e.g. with weak bases, or by treatment with ion exchangers.
  • Salts can be converted in customary manner into the free compounds; metal and ammonium salts can be converted, for example, by treatment with suitable acids, and acid addition salts, for example, by treatment with a suitable basic agent.
  • diastereoisomers can be separated, for example, by partitioning between polyphasic solvent mixtures, recrystallisation and/or chromato ⁇ graphic separation, for example over silica gel, and racemates can be separated, for example, by the formation of salts with optically pure salt-forming reagents and separation of the mixture of diastereoisomers so obtainable, for example by means of fractional crystallisation, or by chromatography over optically active column materials.
  • the present invention relates also to novel starting materials and/or intermediates and to processes for their preparation.
  • the starting materials used and the reaction conditions selected are preferably those that result in the compounds described as being prefe ⁇ ed.
  • starting materials are known, can be prepared according to processes known per se and/or are available commercially.
  • D-, D,L- or L- amino acids, di-, tri- or oligopeptides, derivatized and/or preloaded resins the ancillary reagents and solvents required for either Boc or Fmoc peptide synthesis are commercially available from various suppliers or can be prepared readily according to standard procedures.
  • di-, tri- or oligopeptoids can be prepared readily according to standard procedures.
  • automated peptide synthesizers with optimized, preprogrammed Boc and Fmoc synthesis cycles are available from numerous sources.
  • the compounds of formula V can be prepared by condensation of a carbonic acid of the formula (VI),
  • R 2 ' has the same meaning as R 2 in compounds of formula I or is a resin for solid phase synthesis, while X, B, m and n have the meanings given in the definition of compounds of formula I. If necessary, functional groups that are not to be reacted are present in protected form. The condensation conditions, protective groups, their introduction etc. are analogous to those described for the synthesis of compounds of formula I under process a).
  • H-Nahg-Narg-Narg-Nphe-Narg-NH 2 (SEQ ID NO:2) which is very active in the above-mentioned indications, or a salt thereof.
  • protecting groups in starting materials the reaction of which is to be avoided can be protected by suitable protect ⁇ ing groups (conventional protecting groups) which are customarily used in the synthesis of peptide compounds, and also in the synthesis of cephalosporins and penicillins as well as nucleic acid derivatives and sugars.
  • protect ⁇ ing groups conventional protecting groups
  • These protecting groups may already be present in the precursors and are intended to protect the functional groups in question against undesired secondary reactions, such as acylation, etherification, esterification, oxidation, solvolysis, etc.
  • the protecting groups can additionally cause the reactions to proceed selectively, for example stereoselectively.
  • protecting groups that they can be removed easily, i.e. without undesired secondary reactions taking place, for example by solvolysis, reduction, photolysis, and also enzymatically, for example also under physiological conditions, and, especially, that they are not present in the end products.
  • the protecting groups can be so selected that more than one such group can be removed simultaneously, for example by acidolysis, such as by treatment with trifluoroacetic acid, or with hydrogen and a hydrogenation catalyst, such as a palladium-on-carbon catalyst.
  • the groups can also be so selected that they cannot all be removed simultaneously, but rather in a desired sequence, the co ⁇ esponding inte ⁇ nediates being obtained.
  • any reference hereinbefore and hereinafter to a free compound or a salt thereof is to be understood as meaning also the corresponding salt or free compound, respectively, where appropriate and expedient.
  • All the above-mentioned process steps can be carried out under reaction conditions that are known per se, preferably those mentioned specifically, in the absence or, customarily, in the presence of solvents or diluents, preferably solvents or diluents that are inert towards the reagents used and are solvents therefor, in the absence or presence of catalysts, condensation agents or neutralising agents, for example ion exchangers, such as cation exchangers, e.g.
  • mixtures of isomers that are formed can be separated into the individual isomers, for example diastereoisomers or enantiomers, or into any desired mix ⁇ tures of isomers, for example racemates or mixtures of diastereoisomers, for example ana ⁇ logously to the methods described under "Additional process steps”.
  • solvents from which those solvents that are suitable for any particular reaction may be selected include, for example, water, esters, such as lower alkyl-lower alkanoates, for example ethyl acetate, ethers, such as aliphatic ethers, for example diethyl ether, or cyclic ethers, for example tetrahydrofuran, liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, such as methanol, ethanol or 1- or 2-propanol, nitriles, such as aceto ⁇ nitrile, halogenated hydrocarbons, such as methylene chloride, acid amides, such as dimethylformamide, bases, such as heterocyclic nitrogen bases, for example pyridine, carboxylic acid anhydrides, such as lower alkanoic acid anhydrides, for example acetic anhydride, cyclic, linear or branched hydrocarbons, such as cyclohexane, hexane or iso
  • the compounds, including their salts, may also be obtained in the form of hydrates, or their crystals may, for example, include the solvent used for crystallisation.
  • protected starting materials may be used in all process steps and the protecting groups may be removed at suitable stages of the reaction.
  • the invention relates also to those forms of the process in which a compound obtainable as intermediate at any stage of the process is used as starting material and the remaining process steps are ca ⁇ ied out, or in which a starting material is formed under the reaction conditions or is used in the form of a derivative, for example in protected form or in the form of a salt, or a compound obtainable by the process according to the invention is produced under the process conditions and processed further in situ.
  • a starting material is formed under the reaction conditions or is used in the form of a derivative, for example in protected form or in the form of a salt, or a compound obtainable by the process according to the invention is produced under the process conditions and processed further in situ.
  • reaction conditions that are analogous to those mentioned in the Examples.
  • the invention relates also to pharmaceutical compositions comprising compounds of formula I.
  • the pharmacologically acceptable compounds of the present invention may be used, for example, for the preparation of pharmaceutical compositions that comprise an effective amount of the active ingredient together or in admixture with a significant amount of inorganic or organic, solid or liquid, pharmaceutically acceptable carriers.
  • compositions according to the invention are those for enteral, such as nasal, rectal or oral, or parenteral, such as intramuscular or intravenous, administration to warm-blooded animals (humans and animals), that comprise an effective dose of the pharmacological active ingredient, alone or together with a significant amount of a pharmaceutically acceptable carrier.
  • the dose of the active ingredient depends on the species of warm-blooded animal, the body weight, the age and the individual condition, individual pharmacokinetic data, the disease to be treated and the mode of administration.
  • the invention relates also to a method of treating diseases caused by viruses, especially by retroviral infections, for example HIV infection, including AIDS, which comprises administering a prophylactically or especially therapeutically effective amount of a compound of formula I according to the invention, especially to a warm-blooded animal, for example a human, who on account of one of the mentioned diseases, especially HTV-infection, including AIDS, requires such treatment.
  • the dose to be administered to warm-blooded animals for example humans of approximately 70 kg body weight, is from approximately 3 mg to approximately 3 g, preferably from approximately 10 mg to approximately 1.5 g, for example approximately from 100 mg to 1000 mg per person per day, divided preferably into 1 to 3 single doses which may, for example, be of the same size. Usually, children receive half of the adult dose.
  • compositions comprise from approximately 1 % to approximately 95 %, preferably from approximately 20 % to approximately 90 %, active ingredient.
  • Pharmaceutical compositions according to the invention may be, for example, in unit dose form, such as in the form of ampoules, vials, suppositories, dragees, tablets or capsules.
  • compositions of the present invention are prepared in a manner known per se, for example by means of conventional dissolving, lyophilising, mixing, granulating or confectioning processes.
  • Solutions of the active ingredient, and also suspensions, and especially isotonic aqueous solutions or suspensions are preferably used, it being possible, for example in the case of lyophilised compositions that comprise the active ingredient alone or together with a ca ⁇ ier, for example mannitol, for such solutions or suspensions to be produced prior to use.
  • the pharmaceutical compositions may be sterilised and/or may comprise excipients, for example preservatives, stabilisers, wetting and/or emulsifying agents, solubilisers, salts for regulating the osmotic pressure and/or buffers, and are prepared in a manner known per se, for example by means of conventional dissolving or lyophilising processes.
  • the said solutions or suspensions may comprise viscosity-increasing substances, such as sodium carboxymethylcellulose, carboxymethylcellulose, dextran, polyvinylpy ⁇ olidone or gelatin.
  • Suspensions in oil comprise as the oil component the vegetable, synthetic or semi- synthetic oils customary for injection pu ⁇ oses.
  • liquid fatty acid esters that contain as the acid component a long-chained fatty acid having from 8 to 22, especially from 12 to 22, carbon atoms, for example lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidic acid, behenic acid or co ⁇ esponding unsaturated acids, for example oleic acid, elaidic acid, erucic acid, brasidic acid or linoleic acid, if desired with the addition of anti ⁇ oxidants, for example vitamin E, ⁇ -carotene or 3,5-di-tert-butyl-4-hydroxytoluene.
  • anti ⁇ oxidants for example vitamin E, ⁇ -carotene or 3,5-di-tert-butyl-4-hydroxytoluene.
  • the alcohol component of those fatty acid esters has a maximum of 6 carbon atoms and is a mono- or poly-hydric, for example a mono-, di- or tri-hydric, alcohol, for example methanol, ethanol, propanol, butanol or pentanol or the isomers thereof, but especially glycol and glycerol.
  • fatty acid esters are therefore to be mentioned: ethyl oleate, isopropyl myristate, isopropyl palmitate, "Labrafil M 2375” (polyoxyethylene glycerol trioleate, Gattefosse, Paris), "Miglyol 812” (triglyceride of saturated fatty acids with a chain length of C 8 to C ]2 , H ⁇ ls AG, Germany), but especially vegetable oils, such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and more especially groundnut oil.
  • vegetable oils such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and more especially groundnut oil.
  • the injection compositions are prepared in customary manner under sterile conditions; the same applies also to introducing the compositions into ampoules or vials and sealing the containers.
  • compositions for oral administration can be obtained by combining the active ingredient with solid carriers, if desired granulating a resulting mixture, and processing the mixture, if desired or necessary, after the addition of appropriate excipients, into tablets, dragee cores or capsules. It is also possible for them to be inco ⁇ orated into plastics ca ⁇ iers that allow the active ingredients to diffuse or be released in measured amounts.
  • Suitable carriers are especially fillers, such as sugars, for example lactose, saccharose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, and binders, such as starch pastes using for example corn, wheat, rice or potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or polyvinyl ⁇ py ⁇ olidone, and/or, if desired, disintegrators, such as the above-mentioned starches, also carboxymethyl starch, crosslinked polyvinylpy ⁇ olidone, agar, alginic acid or a salt thereof, such as sodium alginate.
  • fillers such as sugars, for example lactose, saccharose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphat
  • Excipients are especially flow conditioners and lubricants, for example silicic acid, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol.
  • Dragee cores are provided with suitable, optionally enteric, coatings, there being used, inter alia, concentrated sugar solutions which may comprise gum arabic, talc, polyvinylpy ⁇ olidone, polyethylene glycol and/or titanium dioxide, or coating solutions in suitable organic solvents, or, for the preparation of enteric coatings, solutions of suitable cellulose preparations, such as ethylcellulose phthalate or hydroxypropylmethylcellulose phthalate.
  • Capsules are dry-filled capsules made of gelatin and soft sealed capsules made of gelatin and a plasticiser, such as glycerol or sorbitol.
  • the dry-filled capsules may comprise the active ingredient in the form of granules, for example with fillers, such as lactose, binders, such as starches, and/or glidants, such as talc or magnesium stearate, and if desired with stabilisers.
  • the active ingredient is preferably dissolved or suspended in suitable oily excipients, such as fatty oils, paraffin oil or liquid polyethylene glycols, it being possible also for stabilisers and/or antibacterial agents to be added.
  • suitable oily excipients such as fatty oils, paraffin oil or liquid polyethylene glycols, it being possible also for stabilisers and/or antibacterial agents to be added.
  • Dyes or pigments may be added to the tablets or dragee coatings or the capsule casings, for example for identification pu ⁇ oses or to
  • Analytical HPLC is performed on a Shimadzu system (Kyoto, Japan) with product monitoring at 215 n by a Bischoff Lambda 1000 UV-detector from Metrohm (Wallisellen, Switzerland).
  • a C 18 reversed phase column 250 x 4.6 mm, Nucleosil C 18 , 5 ⁇ m; Macherey & Nagel, Duren, Germany) is used. The flow rate is 1 ml/min.
  • the linear gradient is from 10 % B/90 % A to 90 % B/10 % A in 30 min.
  • the analytical data are presented as retention times t R .
  • Matrix-assisted laser deso ⁇ tion ionization (MALDI) mass spectrometry is carried out on a Linear Scientific LDI 1700 (Reno, NV, USA): the sample and 1,5-dihydroxybenzoic acid are co-crystallized, i ⁇ adiated with laser light (337 nm), and the masses of the ions produced are measured in a time-of-flight detection system.
  • MALDI Matrix-assisted laser deso ⁇ tion ionization
  • the Examples 1 to 6 are obtained as trifluoroacetate salts.
  • Example 1 H-Nahg-Narg-Narg-Nphe-Narg-(D-Lvs -(D-LvsV(D-ArgV(D-ProVNH
  • Resin 2 (see example lb)) is washed with ethylene chloride (3 x) and treated with 5 ml 95 % TFA/EDT (8:2) for 15 min. The treatment is repeated twice, while all filtrates are collected and pooled. The resin is washed with DCM (2 x) and TFE (2 x) and the filtrates are added to the pool, which is concentrated to a 5 ml volume in vacuo. 60 ml PE/DIPE (1:1) are added, and the filtrate is isolated and washed with PE/DIPE (1:1). The material is treated with 95% TFA/EDT (8:2) for 120 min, then precipitated again and washed with PE/DIPE (1:1).
  • the material eluted at the main peak is collected, lyophilized and obtained as a white powder.
  • the starting material is prepared as follows: l a) Fmoc-Narg( ' PmcVNarg(Pmc)-Nphe-Narg(Pmc)-D-Lvs(BocVD-Arg(PmcVD-Pro- amide of 4-(2 , ,4'-dimethoxyphenyl-aminomethyl)-phenoxymethyl-polvstyrene resin (Resin 1)
  • the resin is washed 5 times with isopropanol and dried under vacuum.
  • Example 7 (SEQ ID NO:7) Nahg-Narg-Narg-Npeg-Narg-(D-Lys)-NH ? :
  • Resin 1' is washed with ethylene chloride (3x) and treated with 5 ml 95% TFA/EDT (8:2) for 15 min. The treatment is repeated twice, while all filtrates are collected and pooled. The resin is washed with DCM (2x) and TFE (2x) and the filtrates are added to the pool, which is concentrated to a 5 ml volume in vacuo. 60 ml PE/DIPE (1:1) are added and the precipitate is isolated and washed with PE/DIPE (1:1). The material is treated with 95% TFA/EDT (8:2) for 120 min, then again precipitated ans washed with PE/DIPE (1:1). This procedure is repeated with a diminished reaction time of 60 min.
  • the last precipitate is isolated, washed with PE/DIPE (1:1), dissolved in water and lyophilized.
  • the product (title compound) is characterized by analytical HPLC with an analogous chromatography, using a Nucleosil 7C18 (5 ⁇ m) reversed phase column (4.6 x 250 mm, Macherey & Nagel, Duren, FRG), linear gradient 1% B to 90 % B in 30 min; flow 1 ml/min; detection at 215 nm; as well as by mass spectrometry (MALDI on an instrument from Linear Scientific, Reno, NV: model LDI 1700). HPLC-retention time: 11.3 min; MALDI-MS: molecule peak 934 (co ⁇ esponds to theory).
  • the starting material is prepared as follows:
  • MALDI-MS molecule peak 954 (co ⁇ esponds to theory);
  • MALDI-MS molecule peak 992 (corresponds to theory);
  • the starting material is prepared as follows:
  • Boc-protected 2-(4-(2-aminoethyl)phenoxy)ethylbromide is obtained by simple reaction of dibromoethane with Boc-protected 4-(2-aminoethyl)phenol which is obtained easily by Boc-protection of 4-(2-aminoethyl)phenol (Aldrich, Buchs, Switzerland) are reacted with 18.5 g sodium azide at 100 °C during 1 h.
  • the reaction mixture is passed onto alkaline ice water.
  • the suspension is extracted with ether/petrol ether (2 x), and the organic phases are washed with water. Evaporation leads to the title compound, melting point 56-65 °C.
  • MALDI-MS molecule peak 948 (corresponds to theory);
  • the starting material is prepared as follows:
  • the title compound is obtained by catalytic hydrogenation of Boc-protected 2-(amino)-2-(phenyl)-acetonitrile (obtainable by first reacting benzaldehyde cyanhydrin (Aldrich, Buchs, Switzerland) in ethanol with 5 M ammonia at room temperature for 8 days, then evaporation, take up residue in ether, extract with diluted HCl, neutralisation of the extract with NaOH-solution which leads to precipitation, take up product again in ehter and repeat the extraction/precipitation which leads to crystalline 2-(amino)-2-(phenyl)acetonitrile, which is then protected by reaction with Boc-anhydride in tetrahydrofurane) at 105 °C/70 bar in tetrahydrofurane/triethylamine (30:1): melting point 89-92 °C.
  • MALDI-MS molecule peak 1048 (corresponds to theory);
  • the starting material is preapared as follows
  • a sterile-filtered aqueous solution, with 20 % cyclodextrins as solubilisers, of one of the compounds of formula I mentioned in the preceding Examples (e.g. Example 1) as active ingredient, is so mixed under aseptic conditions, with heating, with a sterile gelatine solution containing phenol as preservative, that 1.0 ml of solution has the following composition:
  • Example 13 Sterile dry substance for injection:
  • 500 mg of finely ground ( ⁇ 5.0 ⁇ m) powder of one of the compounds of formula I men ⁇ tioned in the preceding Examples is suspended as active ingredient in a mixture of 3.5 ml of Myglyol 812® and 0.08 g of benzyl alcohol.
  • the suspension is introduced into a container having a metering valve.
  • 5.0 g of Freon 12® are introduced under pressure into the container through the valve.
  • the "Freon” is dissolved in the Myglyol/benzyl alcohol mixture by shaking.
  • the spray container contains approximately 100 single doses which can be administered individually.
  • active ingredient 1000 g corn starch 680 g colloidal silica 200 g magnesium stearate 20 g stearic acid 50 g sodium carboxymethyl starch 250 g water quantum satis
  • a mixture of one of the compounds of formula I mentioned in the preceding Examples (e.g. Example 1) as active ingredient, 50 g of corn starch and the colloidal silica is processed with a starch paste, made from 250 g of corn starch and 2.2 kg of demineralised water, to form a moist mass. This is forced through a sieve having a mesh size of 3 mm and dried at 45° for 30 min in a fluidised bed drier.
  • the dry granules are pressed through a sieve having a mesh size of 1 mm, mixed with a pre-sieved mixture (1 mm sieve) of 330 g of corn starch, the magnesium stearate, the stearic acid and the sodium carboxymethyl starch, and compressed to form slightly biconvex tablets.
  • Xaa Phe Thr Thr Lys Ala Leu Glv lie Ser Tyr Gly Xaa Xaa XaaXaa

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Abstract

Described are peptoid compounds of the formula (I): R1-(A)k-(X)m-(B)n-R2 wherein k, n, R1, R2, A and B have the meanings given in the specification, m is 3 to 10, and any X independently of the others being present represents a bivalent radical of partial formula (II), wherein each of R3, R4 and R5, independently of the others, represents hydrogen or a side chain of an α-amino acid other than glycine, or R3 and R4 together form an alkylene bridge and R5 is hydrogen, or R4 and R5 together form an alkylene bridge and R3 is hydrogen; with the proviso, that a) at least one of these bivalent radicals has an R3 which is a side chain of an α-amino acid other than glycine and proline; b) that at least one R3 other than alkyl or benzyl is present; and c) that the bivalent radicals X together form a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein, and any B independently of the others being present represents a bivalent radical of an α-amino acid, or salts thereof if a salt-forming group is present. The compounds act, for example, as inhibitors of HIV growth by inhibition of the Tat/TAR-interaction.

Description

Antiviral Peptoid Compounds
The present invention relates to antiviral compounds comprising a peptoid structure, processes for the preparation of said compounds, pharmaceutical preparations comprising said compounds, the compounds for the use in the therapeutic (including prophylactic) or diagnostic treatment of the animal or especially human body, and the use of said compounds for the therapeutic or diagnostic treatment of the animal or especially human body or for the manufacture of pharmaceutical preparations.
Background of the invention
The genome of HIV-1, a virus which is regarded as causative agent for the complex disease process leading to AIDS, encodes two regulatory proteins, Tat and Rev, which act through mechanisms the knowledge of which was unprecedented in the scientific community to determine both the quantity and quality of HIV-1 gene expression. These novel regulatory pathways are controlled at the level of protein-RNA interaction. Two classes of HIV mRNAs can be distinguished. The first of these consists of a doubly spliced, 2 kb mRNA species that encodes the viral regulatory proteins, including Tat and Rev. The second class consists of the unspliced (9 kb) and incompletely spliced (4 kb) viral mRNAs that encode the virion structural proteins. Tat induces a marked increase in the steady-state level of viral mRNA. Evidence suggests that this increase can be explained by an increase in the rate of HIV transcription ("transcription booster"). On the other hand, in the absence of Rev, viral RNA transcripts are fully spliced by the cellular RNA processing machinery prior to export to the cytoplasm. In the presence of Rev unspliced (Gag) or partially spliced (Env) viral mRNAs evade processing - instead they are exported directly to the cytoplasm. Rev induces the efficient export of viral RNA species that are otherwise excluded from the cell cytoplasm. Both Tat and Rev recognize regulatory elements on the viral mRNA. Tat function is mediated through a sequence termed TAR (for trans-activation response region) that comprises part of the 5 '-noncoding region of all HIV mRNAs. This region forms a stable stem-loop structure in vitro. Recent evidence indicates that Tat binds directly to the TAR RNA sequence and that this binding is independent of the nucleotide sequence in the loop but dependent on the integrity of the upper stem. The action of Rev protein is highly sequence specific and requires recognition of an RNA target sequence, the Rev Responsive Element (RRE), a highly conserved region in the middle of the viral env gene. The RRE corresponds to a predicted RNA secondary structure of great stability. The RNA binding sites of both Tat and Rev map to protein areas which are highly arginine rich (see Calnan, B.J., et al., Science 252, 1167-1171 (1991) and Tiley, L.S., et al., Proc. Natl. Acad. Sci. USA 89, 758-62 (1992)).
According to estimations by the WHO, more than 20 million people are infected with HIV, a virus, this infection usually leading to the death of the infeted persons due to the development of AIDS and secondary diseases, such as infections, e.g. by Pneumocystis carinii, and/or tumor diseases.
The Tat/TAR interaction has been investigated in more detail in the literature (see Calnan, B.J., et al., Science 252, 1167-1171 (1991) and Calnan, B.J., et al., in: Peptides: Chemistry and Biology, Proc. XII Amer. Peptide Symp., ed. by J.A. Smith and J.E. Rivier, ESCOM Science, Leiden 1992, pp. 685-7). Towards the carboxyl terminus of Tat a highly basic region (residues 48 - 57) is present which appears to be involved in RNA binding. It is of high therapeutic interest to identify new chemical entities with the ability to bind HIV RNA at sites of specific regulatory elements (TAR and RRE), and compete in this process with the viral Tat and Rev proteins. Disruptions of Tat TAR and Rev/RRE complexes corrupt the regulatory systems essential for viral replication and provide a powerful basis for therapeutic intervention in AIDS patients. Different strains of HIV produce different forms of Tat; however, the the N-terminal amino acid sequence of 72 amino acids is common to all forms. The principle form of Tat (designated as Tat(l-86) herein) consists of 86 amino acids in known linear sequence (see Ratner et al., Nature 313, 277 (1985), which is incoφorated by reference herein). Three domains in the protein have been shown to exist by structure/function analysis, including a proline-rich region spanning residues 1-18, a cysteine-rich region spanning residues 22-37, and a basic region of nine amino acid spanning residues 49-57 with the sequence 49(L)-Arg-(L)-Lys-(L)-Lys-(L)-Arg-(L)-Arg-(L)-Gln-(L)-Arg-(L) -Arg-(L)-Arg57,
the latter basic region being designated as "basic domain" of the HIV Tat protein hereinafter.
Summary of the Invention
Surprisingly, it has been found that the compounds of the present invention show very favourable and valuable pharmaceutical characteristics, especially with regard to the therapeutic and/or diagnostic treatment of retroviral infections, particularly AIDS. More specifically, the compounds of the invention comprising a peptoid structure provide synthetic structures with a comparatively low molecular weight which are effective in the treatment of HIV infection, especially HIV-1 infection, acting preferably on the basis of the mechanism described above. The compounds of the invention comprising a peptoid structure represent a class of molecules with the ability to interfere with the Tat/TAR and Rev/RRE complex formations, inter alia with the following distinct advantageous characteristics:
- Protease resistance and useful stability in biological fluids;
- low molecular weight;
- in vivo activity in two different cellular systems at non-cytotoxic concentrations (epitheloid cells co-cultured with constitutively expressing lymphoid cells, as well as human peripheral lymphocytes following an HIV-1 adsoφtion phase); and/or
- the ability to permeate into cells, which can be demonstrated by a Fusion Induced Gene Stimulation Assay that excludes cell surface effects as the underlying mechanism of action.
The compounds of the invention act through a mechanism that provides them with an incomparable therapeutic potential to complement or replace existing, specific or less specific antiviral treatments, with particular value for the treatment against variants of HIV, especially such variants that have become resistant to other kinds of treatment.
Detailed description of the invention
An antiretroviral compound of the invention comprising a peptoid structure is preferably a peptoid compound of the formula I,
R1-(A)k-(X)m-(B)n-R2 (I)
wherein
k is 0 to 20, m is 3 to 10, n is 1 to 10,
R] represents hydrogen, acyl or an amino-substituent other than acyl, R2 represents an OH group, a C-terminal protecting group or a primary, secondary or tertiary amino group,
any A independently of the others being present represents a bivalent radical of an α-amino acid,
any X independently of the others being present represents a bivalent radical of the partial formula II,
Figure imgf000006_0001
wherein
each of R3, R4 and R5, independently of the others, represents hydrogen or a side chain of an α-amino acid other than glycine, or R3 and R4 together form an alkylene bridge and R5 is hydrogen, or R4 and R5 together form an alkylene bridge and R3 is hydrogen;
with the proviso, that a) at least one of these bivalent radicals has an R3 which is a side chain of an α-amino acid other than glycine and proline; b) that at least one R3 other than alkyl or benzyl is present; and c) that the bivalent radicals X together form a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein, and
any B independently of the others being present represents a bivalent radical of an α-amino acid,
or a salt thereof if a salt-forming group is present.
The compounds of the present invention can exist as isomers or mixtures of isomers; for example, if one or more asymmetric carbon atoms are present, these carbon atoms can be in the (R)-, (S)- or (R,S)-configuration, independent of one another. It is thus possible to obtain isomeric mixtures, such as racemates or diastereomeric mixtures, or pure diastereomers or entantiomers, depending on the number of asymmetric carbon atoms and on whether isomers or isomeric mixtures are present. Preferred are pure isomers (enantiomers or diastereomers).
Unless otherwise indicated, the general terms and names used in the description of the present invention preferably have the following meanings:
In compounds of formula I, k is 0 to 20, preferably 0 to 12, most preferably 0 or 12; m is 3 to 10, preferably 4 to 6, most preferably 5; and n is 1 to 10, preferably 1 to 4.
The term "lower" defines a moiety with up to and including maximally 7, especially up to and including maximally 4, carbon atoms, said moiety being branched or straight-chained. Lower alkyl, for example, is methyl, ethyl, n-propyl, sec-propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl or n-heptyl.
Acyl has, for example, up to 25, preferably up to 19, carbon atoms and is especially the acyl group of a carboxylic acid or of a semiester of carbonic acid.
Preferred acyl groups of a carboxylic acid are unsubstituted or substituted alkanoyl having up to 19 carbon atoms, for example n-decanoyl, or preferably
lower alkanoyl, such as formyl, acetyl, propionyl, butyryl or pivaloyl; or
substituted lower alkanoyl wherein preferably up to four, especially (except in the case of halogen which may be present up to three times as a substituent) up to two, substituents may be present, especially one substituent (except in the case of halogen which may be present up to three times as a substituent), the substituents being independently selected especially from
cycloalkyi with from 3 to 7 carbon atoms, especially in cycloalkyl-lower alkanoyl wherein lower alkanoyl is as defined above, for example cycloalkylcarbonyl, such as cyclopropyl-, cyclobutyl-, cyclopentyl- or cyclohexyl-carbonyl, or 2-cyclohexyl- or 2-cyclopentyl-acetyl,
aryl which has preferably from 6 to 14 ring carbon atoms, such as in phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, and may be unsubsti¬ tuted or mono- to tri-substituted especially by lower alkyl, for example methyl, ethyl or propyl, halo-lower alkyl, for example trifluoromethyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, for example methoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl- lower alkoxy or N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoyl amino, halogen, for example fluorine, chlorine or bromine, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, such as benzyloxycarbonyl, lower alkanoyl, sulfo, lower alkanesulfonyl, for example methanesulfonyl (CH3-S(O)2-), phosphono (-P(=O)(OH)2), hydroxy-lower alkoxy- phosphoryl or di-lower alkoxyphosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro and/or by cyano,
carbamoyl,
carbamoyl substituted at the nitrogen atom by one or two radicals selected from lower alkyl, carboxy-lower alkyl or lower alkoxycarbonyl-lower alkyl,
heterocyclyl which is preferably a single or double ring system having from 3 to 10 ring atoms, is bonded via a carbon atom or, especially, via a nitrogen atom and contains up to 3 further hetero atoms selected from oxygen, nitrogen, sulfur, and sulfur linked to 1 or 2 oxygen atoms; which in addition may also be fused with 1 or 2 phenyl radicals or with 1 or 2 cycloalkyi radicals, cycloalkyi preferably having from 5 to 7 ring atoms; and which may be unsaturated or partially or fully saturated, for example thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, chromanyl, cyclohexa[b]pyrrolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa- [b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S-dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydro- indolyl, 1, 2,3,4- tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, with heterocyclyl, for example one of the last-mentioned radicals, being unsubstituted or substituted by one or more substituents selected from lower alkyl, for example methyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, for example benzyl, hydroxy-lower alkyl, for example hydroxymethyl or 2-hydroxyethyl, hydroxy, lower alkoxy, for example methoxy or ethoxy, amino, lower alkylamino, for example methyl-, ethyl- or tert-butyl-amino, di-lower alkylamino, for example dimethyl- or diethyl-amino, carboxy, lower alkoxycarbonyl, for example methoxy-, isopropoxy-, sec-butoxy- or tert-butoxy-carbonyl, phenyl- or naphthyl-lower alkoxycarbonyl, for example benzyloxy carbonyl, halogen, for example fluorine, chlorine, bromine or iodine, especially chlorine or bromine, lower alkanoyl, for example acetyl or pivaloyl, nitro, oxo and/or by cyano;
hydroxy, especially in hydroxy-lower alkanoyl,
lower alkoxy, especially in lower alkoxy-lower alkanoyl, for example lower alkoxy- acetyl or lower alkoxypropionyl, such as methoxyacetyl, ethoxyacetyl or 3-methoxypropionyl,
lower alkanoyloxy, especially in lower alkanoyloxy-lower alkanoyl
up to 3 halogen atoms, especially in halo-lower alkanoyl containing up to 3 halogen atoms, for example α-haloacetyl, such as α-fluoro-, α-chloro-, α-bromo-, α-iodo-, α,α,α-trifluoro- or α,α,α-trichloro-acetyl, or halopropionyl, such as β-chloro- or β- bromo-propionyl, and
carboxy, especially in carboxy-lower alkanoyl.
The above-mentioned general definitions of the terms "aryl", "cycloalkyi" and "heterocyclyl" are not restricted to the substitutents of lower alkanoyl, but are generally valid where these terms appear in the present specification, subject to the specific definitions of those moieties that, in each case, are characterized as being preferred.
A preferred acyl group of a semiester of carbonic acid has up to 19 carbon atoms and is selected preferably from the group comprising
lower alkoxycarbonyl, for example methoxy-, ethoxy-, n-propoxy-, isopropoxy-, isobutoxy- or tert-lower alkoxy-carbonyl, or also or especially n-propoxycarbonyl, such as tert-butoxycarbonyl or isobutoxycarbonyl,
2-halo-lower alkoxycarbonyl, such as 2-chloro-, 2-bromo-, 2-iodo- or 2,2,2-trichloro- ethoxycarbonyl, aryl-lower alkoxycarbonyl, for example arylmethoxy-carbonyl, wherein aryl has from 6 to 14 carbon atoms and is, for example, phenyl, 1- or 2-naphthyl, fluorenyl, especially 9-fluorenyl, or phenyl mono- or poly-substituted by lower alkyl, for example methyl or tert-butyl, phenyl, hydroxy, lower alkoxy, for example methoxy, ethoxy or tert-butoxy, halogen, for example chlorine or bromine, and/or by nitro, for example phenyl-lower alkoxycarbonyl, such as benzyloxy carbonyl, 4-nitrobenzyloxycarbonyl, diphenylmethoxycarbonyl or 4-methoxybenzyloxycarbonyl,
heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is preferably a single or double ring system having from 3 to 10 ring atoms, is bonded via a carbon atom or, especially, via a nitrogen atom and contains up to 3 further hetero atoms selected from oxygen, nitrogen, sulfur, and sulfur linked to 1 or 2 oxygen atoms; which in addition may also be fused with 1 or 2 phenyl radicals or with 1 or 2 cycloalkyi radicals, cycloalkyi preferably having from 5 to 7 ring atoms; and which may be unsaturated or partially or fully saturated, for example thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, cyclohexa[b]pyrrolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S-dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4- tetrahydroisoquinolyl, with heterocyclyl, for example one of the last-mentioned radicals, being unsubstituted or substituted by lower alkyl, for example methyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, for example benzyl, hydroxy-lower alkyl, for example hydroxymethyl or 2-hydroxyethyl, hydroxy, lower alkoxy, for example methoxy or ethoxy, amino, lower alkylamino, for example methyl-, ethyl- or tert-butyl-amino, di-lower alkylamino, for example dimethyl- or diethyl-amino, carboxy, lower alkoxycarbonyl, for example methoxy-, isopropoxy-, sec-butoxy- or tert-butoxy-carbonyl, phenyl- or naphthyl-lower alkoxycarbonyl, for example benzyloxycarbonyl, halogen, for example fluorine, chlorine, bromine or iodine, especially chlorine or bromine, lower alkanoyl, for example acetyl or pivaloyl, nitro, oxo and/or by cyano;
lower alkenyloxycarbonyl wherein preferably the lower alkenyl radical is bonded to the bonding oxygen atom via a saturated carbon atom, such as allyloxycarbonyl,
lower alkoxy-lower alkoxycarbonyl, such as 2-methoxyethoxycarbonyl, and (lower alkoxy-lower alkoxy )-lower alkoxycarbonyl, such as 2-(2-methoxyethoxy)- ethoxycarbonyl.
An amino-substituent other than acyl is preferably an arylmethyl, etherified mercapto, 2-acyl-lower alk-1-enyl, a silyl group or an organic sulfonyl group.
In arylmethyl, for example a mono-, di- or especially tri-arylmethylgroup, each aryl radical has preferably from 6 to 14 ring carbon atoms, such as in phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, and may be unsubstituted or mono- to tri-substituted especially by lower alkyl, for example methyl, ethyl or propyl, halo-lower alkyl, for example trifluoromethyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, for example methoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy or N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkyl¬ amino, lower alkanoylamino, halogen, for example fluorine, chlorine or bromine, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, such as benzyloxycarbonyl, lower alkanoyl, sulfo, lower alkanesulfonyl, for example methanesulfonyl (CH3-S(O)2-), phosphono (-P(=O)(OH)2), hydroxy-lower alkoxy¬ phosphoryl or di-lower alkoxyphosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro and/or by cyano; such groups are, for example, benzyl, diphenylmethyl or especially trityl.
In etherified mercapto the mercapto group is especially in the form of substituted arylthio or aryl-lower alkylthio, wherein aryl is, for example, phenyl that is unsubstituted or substituted, for example, by lower alkyl, such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen, such as chlorine, and/or by nitro, for example 4-nitrophenylthio (= 4-nitrophenylsulf enyl) .
In 2-acyl-lower alk-1-enyl, acyl is, preferably, the corresponding radical of a lower alkanoic acid, of a benzoic acid that is unsubstituted or substituted, for example, by lower alkyl, such as methyl or tert-butyl, lower alkoxy, such as methoxy, halogen, such as chlorine, and/or by nitro, or especially of a carbonic acid semiester, such as a carbonic acid lower alkyl semiester. Corresponding groups are especially 1-lower alkanoyl-lower alk-l-en-2-yl, for example 1-lower alkanoylprop-l-en-2-yl, such as 1-acetyl- prop-l-en-2-yl, or lower alkoxycarbonyl-lower alk-l-en-2-yl, for example lower alkoxycarbonylprop-l-en-2-yl, such as l-ethoxycarbonylprop-l-en-2-yl. A siiyl group is, for example, a tri-lower alkylsilyl group, for example trimethylsilyl or tert-butyl-dimethylsilyl.
An organic sulfonyl group is preferably selected from the group comprising arylsulfonyl, wherein aryl is preferably phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups, such as 4-methylphenylsulfonyl (= tosyl), (mono-, di- or tri-aryl)-lower alkylsulfonyl, wherein aryl is preferably phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups, such as benzylsulfonyl, 4-methoxy-2,3,6-trimethylbenzylsulfonyl (Mtr) or 4-methylbenzylsulfonyl, and hetero- cyclyl-sulfonyl, such as 2,2,5,7,8-pentamethylchroman-6-sulfonyl or 2,2,4,6,7-penta- methyldihydrobenzofuran-5-sulfonyl (Pbf).
A C-terminal protecting group is preferably an esterifying group, thus leading to an esterified C-terminal carboxy group. More preferred is a lower alkoxy group that is preferably branched in the 1 -position of the lower alkoxy group or substituted in the 1- or 2-position of the lower alkoxy group by suitable substituents.
A lower alkoxy group that is branched in the 1 -position of the lower alkoxy group is, for example, tert-lower alkoxy, for example tert-butoxy.
A lower alkoxy group that is substituted in the 1- or 2-position of the lower alkoxy group by suitable substituents is, for example, arylmethoxy having one or two aryl radicals, wherein aryl is preferably phenyl that is unsubstituted or mono-, di- or tri-substituted, for example, by lower alkyl, for example tert-lower alkyl, such as tert-butyl, lower alkoxy, for example methoxy, hydroxy, halogen, for example chlorine, and/or by nitro, for example benzyloxy, benzyloxy substituted by the mentioned substituents, for example 4-nitro- benzyloxy or 4-methoxybenzyloxy, diphenylmethoxy or diphenylmethoxy substituted by the mentioned substituents, for example di(4-methoxyphenyl)methoxy; 1-lower alkoxy- lower alkoxy, for example methoxymethoxy, 1 -methoxy ethoxy or 1-ethoxyethoxy, 1-lower alkylthio-lower alkoxy, for example 1-methylthiomethoxyl or 1-ethylthioethoxy, aroylmethoxy wherein the aroyl group is preferably benzoyl that is unsubstituted or substituted, for example, by halogen, such as bromine, for example phenacyloxy, 2-halo- lower alkoxy, for example 2,2,2-trichloroethoxy, 2-bromoethoxy or 2-iodoethoxy, as well as 2-(tri-substituted silyl)-lower alkoxy wherein the substituents are each independently of the others selected from lower alkyl, phenyl-lower alkyl, cycloalkyi or phenyl each of which is unsubstituted or substituted as above, for example 2-tri-lower alkylsilyl-lower alkoxy, such as 2-tri-lower alkylsilylethoxy, for example 2-trimethylsilylethoxy or 2-(di-n-butyl-methyl-siiyl)-ethoxy, or triphenylsilylethoxy.
A C-terminal protecting group can furthermore be an organic silyloxy group. An organic silyloxy group is, for example, a tri-lower alkylsilyloxy group, for example trimethylsilyloxy. The silicon atom of the silyloxy group can also be substituted by two lower alkyl groups, for example methyl groups.
A C-terminal protecting group is preferably tert-lower alkoxy, for example tert-butyloxy, benzyloxy, 4-nitrobenzyloxy, 9-fluorenylmethoxy or diphenylmethoxy.
A primary, secondary or tertiary amino group is preferably a free amino group, a mono- or disubstituted amino group the substituents of which are preferably selected from the group comprising lower alkyl, e.g. methyl or ethyl, aryl-lower alkyl, such as phenyl-lower alkyl, e.g. benzyl, or heterocyclyl-lower alkyl, such as pyrrolidinyl-lower alkyl, e.g. 2-(l-pyrrolidinyl)-ethyl, pyridyl-lower alkyl, e.g. 2-(2-pyridyl)-ethyl, furyl-lower alkyl, e.g. 2-furylmethyl, moφholinyl-lower alkyl, e.g. 2-(4-moφholinyl)-ethyl, and indolyl-lower alkyl, e.g. 2-(3-indolyl)-ethyl. A disubstituted amino group may also be N-containing heterocyclyl bonded via its nitrogen atom, such as e.g. 1-pyrrolidinyl or 4-moφholinyl.
A bivalent radical of an α-amino acid is preferably bonded N-terminally by way of its α-amino group and C-terminally by way of its carboxy group and is preferably selected from the group comprising a bivalent radical of a natural α-amino acid having the L-confϊguration, such as those normally occurring in proteins, or an epimer of such an amino acid, that is to say having the unnatural D-configuration, or a D,L-isomeric mixture thereof; or a homologue of such an amino acid, for example wherein the amino acid side chain has been shortened by one or two methylene groups or lengthened to up to 10 carbon atoms, such as an α-amino alkanoic acid with 5 up to and including 10 carbon atoms in a linear chain, a substituted aromatic (α-aryl or α-aryl lower alkyl) amino acid wherein the aryl radical has from 6 to 14 carbon atoms, for example a substituted phenylalanine or phenylglycine wherein the phenyl may be mono- or poly-substituted by lower alkyl, for example methyl, lower alkoxy, for example methoxy, lower alkanoyloxy, for example acetoxy, amino, lower alkylamino, for example methylamino, di-lower alkylamino, for example dimethylamino, lower alkanoylamino, for example acetylamino or pivaloyl- amino, lower alkoxycarbonylamino, for example tert-butoxycarbonylamino, arylmethoxy- carbonylamino wherein aryl preferably has from 6 to 14 carbon atoms, for example benzyloxycarbonylamino or 9-fluorenylmethoxycarbonylamino, halogen, for example fluorine, chlorine, bromine or iodine, carboxy and/or by nitro, a benzo-fused phenyl¬ alanine or phenylglycine, such as α-naphthylalanine, and a hydrogenated phenylalanine or phenylglycine, such as cyclohexylalanine or cyclohexylglycine, or an α-amino heterocyclyl-lower alkanoic acid wherein heterocyclyl preferably is a single or double ring system having from 3 to 10 ring atoms, is bonded via a carbon atom or via a nitrogen atom and contains up to 3 further hetero atoms selected from oxygen, nitrogen, sulfur, and sulfur linked to 1 or 2 oxygen atoms, and may be unsaturated or partially or fully saturated, for example furyl, pyrrolyl, pyrrolidinyl, moφholinyl, pyridyl or indolyl, said homologue being present in the L-, D- or (D,L)-configuration.
Especially preferred is the bivalent radical, bonded via its α-amino and its α-carbonyl group, of an amino acid selected from glycine (H-Gly-OH), alanine (H-Ala-OH), valine (H-Val-OH), norvaline (α-aminovaleric acid), leucine (H-Leu-OH), isoleucine (H-Ile-OH), norleucine (α-aminohexanoic acid, H-Nle-OH), α-amino-n-decanoic acid, serine (H-Ser-OH), homoserine (α-amino-γ-hydroxybutyric acid), threonine (H-Thr-OH), methionine (H-Met-OH), cysteine (H-Cys-OH), proline (H-Pro-OH), trans-3- and trans-4-hydroxyproline, phenylalanine (H-Phe-OH), tyrosine (H-Tyr-OH), 4-amino- phenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenylalanine, β-phenylserine (β-hydroxyphenylalanine), phenylglycine, α-naphthylalanine (H-Nal-OH), cyclohexylalanine (H-Cha-OH), cyclohexylglycine, tryptophan (H-Tφ-OH), indoline- 2-carboxylic acid, l,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aspartic acid (H-Asp-OH), asparagine (H-Asn-OH), aminomalonic acid, aminomalonic acid mono- amide, glutamic acid (H-Glu-OH), glutamine (H-Gln-OH), histidine (H-His-OH), arginine (H-Arg-OH), lysine (H-Lys-OH), Ne-benzyl-N€-methyl-lysine, Ne,Ne-dibenzyl-lysine, δ- hydroxylysine, ornithine (α,δ-diaminovaleric acid), α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyrrolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino- γ-(4-moφholinyl), α-amino-γ-(3-indolyl)-butyric acid, α,γ-diaminobutyric acid and α,β- diaminopropionic acid, especially preferably the bivalent radical of an α-amino acid, bonded via its α-amino and its α-carbonyl group, selected from glycine, alanine, valine, leucine, isoleucine, α-amino-decanoic acid, serine, threonine, proline, phenylalanine, tyrosine, 4-nitrophenylalanine, glutamic acid, arginine, lysine,
Ne-benzyl-Ne-methyl-lysine, Ne,Ne-dibenzyI-lysine, α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyrrolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino- γ-(4-moφholinyl)-butyric acid and α-amino-γ-(3-indolyl)-butyric acid;
it being possible for each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form, preferably in the L- or in the D-form.
In a bivalent radical of the partial formula II, a side chain of an α-amino acid other than glycine and proline is the group other than the amino and the carboxy group bound to the α-carbon of the respective amino acid, that is a group R6 that is bound to an amino acid structure of the formula III
Figure imgf000015_0001
wherein R6 is a side chain selected so as to give an α-amino acid (except for glycine where the residue corresponding to R6 would be H and except for proline), more specifically one of the α-amino acids as defined above; more preferably, R5 is a residue from the group comprising lower alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or sec-butyl, or substituted lower alkyl, such as aryl-lower alkyl wherein the aryl is preferably phenyl that may be mono- or poly-substituted by lower alkyl, for example methyl, lower alkoxy, for example methoxy, lower alkanoyloxy, for example acetoxy, amino, lower alkylamino, for example methylamino, di-lower alkylamino, for example dimethylamino, lower alkanoylamino, for example acetylamino or pivaloylamino, lower alkoxycarbonylamino, for example tert-butoxycarbonylamino, arylmethoxycarbonylamino wherein aryl preferably has from 6 to 14 carbon atoms, for example benzyloxycar- bonylamino or 9-fluorenylmefhoxycarbonylamino, halogen, for example fluorine, chlorine, bromine or iodine, carboxy and/or by nitro, for example benzyl or 4-nitrobenzyl; (basic) amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl; (basic) N-lower alkylamino-, such as N-methylamino-, (basic) N,N-di-lower alkylamino-, (basic) N-(aryl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, (basic) N,N-di(aryl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or (basic) guanidino-lower alkyl (lower alkyl preferably being methyl, ethyl or propyl), such as 3-guanidinopropyl; carboxy-lower alkyl, such as 2-carboxyethyl, carbamoyl-lower alkyl (H2N-C(=O)-lower alkyl), such as 2-carbamoylethyl or 3-carbamoylρropyl, and heterocyclyl-lower alkyl, especially (basic) pyridyl-lower alkyl, for example (2-pyridyl)-lower alkyl, such as 2-(2-pyridyl)ethyl, (basic) pyrrolidinyl-lower alkyl, for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pyrrolidinyl)ethyl, furyl-lower alkyl, for example (2-furyl)-lower alkyl, such as (2-furyl)methyl, (basic) moφholinyl-lower alkyl, for example (4-moφholinyl)- lower alkyl, such as 2-(4-moφholinyl)ethyl, and indolyl-lower alkyl, for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl; or (alternatively to the above- mentioned residues or included in their group) selected from (basic) (acridin-9-yl)- amino-lower alkyl, (3,6-bis(dimethylamino)acridin-9-yl)amino-lower alkyl or (6-chloro- 9-methoxyacridin-9-yl)amino-lower alkyl, such as 2-(acridin-9-yl)amino-ethyl, 2-(3,6-bis(dimethylamino)acridin-9-yl)amino-ethyl or 2-(6-chloro-9-methoxyacri- din-9-yl)amino-ethyl, (basic) (amino)-(phenyl)-lower alkyl, such as 2-amino-2-phenyl- ethyl, (basic) (amino-loweralkylphenoxy)-lower alkyl, such as 2-(4-(2-aminoethyl)- phenoxy)-ethyl, and (basic) oxopyrrolidinyl-lower alkyl, such as 3-(2-oxopyrrolidin- l-yl)propyl.
Throughout this application, at least one bivalent residue of the partial formula II mst be present wherein R3 is other than alkyl and benzyl, any other moieties R3 if present being allowed to be lower alkyl or benzyl in a compound of formula I. The term alkyl preferably means lower alkyl, especially CrC4-alkyl, such a methyl or ethyl.
An alkylene bridge formed by R3 and R4 together, or by R4 and R5 together, preferably is a lower alkylene bridge, such as -(CH2)3-, -(CH2)4- or -(CH^-, most preferably -(CH2) - (so that the respective bivalent radical is a prolyl radical).
The term "comprising a peptoid structure" means that the compounds of formula I are oligoamidic compounds with at least one residue X of formula II in formula I wherein R3 is not hydrogen, but a side chain of an amino acid other than glycine and proline.
TAR-binding perferably means binding as measured in the Tat-TAR gel-shift assay described below.
For radicals X that together form a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein, preferably, in this analogue m is 4 to 9, preferably 4 to 6; and the basic side chains R3, R4 and/or R5 mentioned in the last paragraph above are preferred, with the exception of the X at position 4 of the residue -(X)m- (counted from the N-terminus, that is the underlined moiety in the following representation of the residue -(X)m-, namely -X-X-X-X- ...) where any of the side chains mentioned above, especially in the last paragraph, is possible; and more preferably
each X is a bivalent radical of the partial formula II,
Figure imgf000017_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or sec-butyl, or substituted lower alkyl, such as aryl-lower alkyl wherein the aryl is preferably phenyl that may be mono- or poly-substituted by lower alkyl, for example methyl, lower alkoxy, for example methoxy, lower alkanoyloxy, for example acetoxy, amino, lower alkylamino, for example methylamino, di-lower alkylamino, for example dimethylamino, lower alkanoylamino, for example acetylamino or pivaloylamino, lower alkoxycarbonylamino, for example tert-butoxycarbonylamino, arylmethoxycarbonylamino wherein aryl preferably has from 6 to 14 carbon atoms, for example benzyloxycarbonylamino or 9-fluorenylmethoxy- carbonylamino, halogen, for example fluorine, chlorine, bromine or iodine, carboxy and/or by nitro, for example benzyl or 4-nitrobenzyl; (basic) amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl; (basic) N-lower alkylamino-, such as N-methylamino-, (basic) N,N-di-lower alkylamino-, (basic) N-(aryl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, (basic) N,N-di(aryl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or (basic) guanidino-lower alkyl (lower alkyl preferably being methyl, ethyl or propyl), such as 3-guanidinopropyl; carboxy-lower alkyl, such as 2-carboxyethyl, carbamoyl-lower alkyl (H2N-C(=O)-lower alkyl), such as 2-carbamoylefhyl or 3-carbamoylpropyl, and heterocyclyl-lower alkyl, especially (basic) pyridyl-lower alkyl, for example (2-pyridyl)-lower alkyl, such as 2-(2-pyridyl)ethyl, (basic) pyrrolidinyl-lower alkyl, for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pyrrolidinyl)ethyl, furyl-lower alkyl, for example (2-furyl)-lower alkyl, such as (2-furyl)mefhyl, (basic) moφholinyl-lower alkyl, for example (4-moφholinyl)-lower alkyl, such as 2-(4-moφholinyl)ethyl, or indolyl-lower alkyl, for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl; with the proviso that with the exception of the X at position 4 of the residue -(X)m- (counted from the N-terminus, that is the underlined moiety in the following representation of the residue -(X)m-> namely -X-X-X-X- ...) where any of the mentioned side chains R3 mentioned above is possible, the side chain R3 of any X (in a different than the 4-position) is selected from those moieties characterized above as "basic", and m is 4 to 6, preferably 5;
and wherein
each of R4 and R5 represents hydrogen.
In the definition of the radical A, a bivalent radical of an α-amino acid is preferably a bivalent radical of an amino acid selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present preferably in the L-form (where an asymmetric α-carbon atom is present).
In the definition of the radical B, a bivalent radical of an α-amino acid is preferably a bivalent radical of an amino acid selected from lysine, arginine and proline, each of which is present preferably in the D-form.
Preferably, A and B are selected so as not to interfere with the binding of the respective compound of formula I to the TAR target. Most preferred are sequences that are analogous (with small deviations, such as conservative substitutions of amino acids, e.g. up to three such substitutions ) or identical to those in the corresponding Tat (1-86).
Salts of compounds of formula I are especially acid addition salts, salts with bases or, where several salt-forming groups are present, can also be mixed salts or internal salts.
Salts are especially pharmaceutically acceptable salts of compounds of formula I.
Such salts are formed, for example, from compounds of formula I having an acid group, for example a carboxy group, a sulfo group, or a phosphoryl group substituted by one or two hydroxy groups, and are, for example, salts thereof with suitable bases, such as non-toxic metal salts derived from metals of groups Ia, lb, Ila and lib of the Periodic Table of the Elements, especially suitable alkali metal salts, for example lithium, sodium or potassium salts, or alkaline earth metal salts, for example magnesium or calcium salts, also zinc salts or ammonium salts, as well as salts formed with organic amines, such as unsubstituted or hydroxy-substituted mono-, di- or tri-alkylamines, especially mono-, di- or tri-lower alkylamines, or with quaternary ammonium compounds, for example with N-methyl-N-ethylamine, diethylamine, triethylamine, mono-, bis- or tris-(2-hydroxy-lower alkyl)amines, such as mono-, bis- or tris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butyl- amine or tris(hydroxymethyl)methylamine, N,N-di-lower alkyl-N-(hydroxy-lower alkyl¬ amines, such as N,N-dimethyl-N-(2-hydroxyethyl)-amine or tri-(2-hydroxyethyl)-amine, N-methyl-D-glucamine, or quaternary ammonium salts, such as tetrabutylammonium salts. The compounds of formula I having a basic group, for example an amino group, can form acid addition salts, for example with inorganic acids, for example hydrohalic acids, such as hydrochloric acid, sulfuric acid or phosphoric acid, or with organic carboxylic, sulfonic, sulfo or phospho acids or N-substituted sulfamic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid, methyl- maleic acid, fumaric acid, malic acid, tartaric acid, gluconic acid, glucaric acid, glucuronic acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-aminosali- cylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, as well as with amino acids, for example the α-amino acids mentioned hereinbefore, especially glutamic acid and aspartic acid, and with methanesulfonic acid, ethanesulfonic acid, 2-hydroxy ethanesulfonic acid, ethane- 1,2-disulfonic acid, benzene- sulfonic acid, 4-methylbenzenesulfonic acid, naphthalene- 2-sulfonic acid, 2- or 3-phosphoglycerate, glucose-6-phosphate, N-cyclohexylsulfamic acid (forming cycla- mates) or with other acidic organic compounds, such as ascorbic acid. Compounds of formula I having acid and basic groups can also form internal salts.
For isolation or purification puφoses, it is also possible to use pharmaceutically unaccept¬ able salts.
The compounds of the invention have useful, in particular pharmacologically useful, properties. Suφrisingly, it has been found that the compounds of formula I are able to inhibit the propagation of HIV, especially HIV-1, in infected human lymphocytes and show a particularly potent, specific inhibition on the binding of the Tat protein to TAR, mainly by binding to TAR. They thus represent a totally new class of inhibitors and therapeutics.
The in vitro inhibition of the interaction between Tat and TAR can be shown by a competition Tat-TAR gel-shift assay. Through binding of the protein (recombinant Tat, Medical Research Council (MRC), Cambridge, U.K. - the sequence of recombinant Tat can be found in Churcher et al., J. Mol. Biol. 230, 90-110 (1993) to the RNA (synthetic TAR duplex, Genset, Paris, France; the sequence can be found in Hamy et al., J. Mol. Biol. 230, 111-123 (1993)), the overall size and the charge/weight ratio of the formed duplex are changed, so that electrophoretic migration through a native polyacrylamide gel is affected. By radioactive labelling of the RNA and subsequent autoradiography, free RNA and complexes can be discriminated based on their relative positions in the gel (Hamy et al., J. Mol. Biol. 230, 111-123 (1993)). If the binding reaction with a substance able to prevent the protein binding to the radiolabelled RNA, this competition for binding can be visualized on the autoradiography as a decreased intensity of the retarded band. In more detail, compounds of the formula I are tested as follows:
The 14-mer strand of duplex TAR-RNA is labelled in the presence of T4 polynucleotide kinase (New England Biolabs, Beverly, MA, USA) using [γ-32P]ATP (Amersham, Little Chalfont, UK), 10 mM DTT (=dithiothreitol), 50 mM Tris-HCl (pH 7.4, Tris = Tris(hydroxymethyl)aminomethane), 0.77 % (w/v) spermidine (Fluka, Switzerland) and 10 units T4 polynucleotide kinase incubated at 37 °C for 20 min. After heat-treatment (65 °C, 10 min) for enzyme inactivation, the unincoφorated [γ-32P]ATP is removed by chromatography through a sephadex NAP- 10 column (Pharmacia, Uppsala, Sweden) equilibrated with water. Prior to the in-vitro-binding reaction, the labelled 14-mer is annealed to 1.5 equivalents of unlabelled 17-mer by heating to 90 °C for 3 min, followed by slow cooling down to 0 °C. The binding reaction for protein and RNA which takes place in a volume of 25 μl contains approximately 10,000 cpm of the labelled duplex TAR-RNA and 20 nM recombinant Tat protein in TK buffer (Tris-HCl 20 mM pH 8.0, KCl 50 mM) with 10 mM DTT, 0.1 % Triton X-100 ((Alkylphenylpolyethylenglykol, Rohm & Haas, Darmstadt, Germany) in the absence or presence of varying concentrations of inhibitor. The autoradiographies are quantified by Phosphorimager (Molecular Dynamics/Bucher, Basle, Switzerland). A CD50 value is determined as the concentration of a compound of the formula I giving a 50 % decrease in the intensity of the retarded band (Tat-TAR complex). The CD50-values that are obtained are preferably in the range of from 1 x 10"9 to 1 x 10"6 M. It is possible to show that similar binding affinity can be found when wild-type unlabelled TAR-RNA is used as competitor, thus suggesting that the compounds of the present invention have affinities comparable to that of the high molecular weight full-length Tat protein in vitro.
In close analogy, the compounds of the present invention can also be shown to inhibit Rev/RRE interaction in a competition Rev-RRE gel-shift assay (see Kjems, J., et al., EMBO J. 11, 1119-1129 (1992) and Tan, R., et al., Cell 73, 1031-40 (1993).
In addition, the inhibition of viral growth of HIV-1 in cellular systems in vitro can be demonstrated by procedures known in the art, e.g. according to the method described in Lazdins et al., AIDS Research and Human Retroviruses 8(4), 505 -11 (1992). In brief, Peripheral Blood Mononuclear Lymphocytes (PBLs) are obtained from the blood of healthy volunteers by leukapheresis. Cells (1 x 106/ml) are cultured for 2 days in RPMI-1640 (Gibco), supplemented with 10 % heat-inactivated fetal calf serum (Gibco), 50 μg ml streptomycin, 50 U/ml penicillin (Amimed), 2 nM glutamine and 10 mM hepes buffer (Gibco). Stimulated lymphocytes are obtained by culturing in the presence of PHA (0.25 μg/ml; Wellcome diagnostics, Templehill, Dartford, England). PHA-lymphocyte stimulation is confirmed by the increase in cell size (Scattergram, FACS analysis). Cells are exposed to HIV-1/LAV.04 (see Barre-Sinoussi et al., Science 220, 868-871 (1983)) for 6 h. Following infection, the cells are washed and resuspended in RPMI-1640 (see above) supplemented with human IL-2 (Genzyme, Cambridge, MA) at 0.6 x 106/ml. Medium is changed every 3 days and activity of viral reverse transcriptase (RT activity) is determined in cell supernatants, serving as a measure for the presence of virus and thus of progression of infection according to known procedures (see Willey et al., J. Virol. 62, 353-8 (1991) for details of the method). In brief, RT determination is possible as follows: The RT activity is determined in 50 mM of tris (α,α,α-tris(hydroxymethyl)methylamine, ultra pure, Merck, Federal Republic of Germany) pH 7.8; 75 mM of KCl, 2 mM of dithiothreitol, 5 mM of MgCl2; 0.05 % Nonidet P-40 (detergent; Sigma, Switzerland); 50 μg/ml of polyadenylic acid (Pharmacia, Sweden); 1.6 μg/ml of dT(12-18) (Sigma, Switzerland). The mixture is filtered through an Acrodisc filter (0.45 μ: Gellman Science Inc, Ann Arbor) and stored at -20°C. 0.1% (v/v) [alpha-32P]dTTP is added to aliquots of that solution in order to achieve a final radioactive activity of 10 μCi/ml. 10 μl of the culture supernatant are transferred to a new 96-well microtitre plate and 30 μl of the mentioned RT cocktail are added thereto. After mixing, the plate is incubated for from 1.5 to 3 hours at 37°C. 5 μl of that reaction mixture are transferred to Whatman DE81 paper (Whatman, Maidstone, UK). The dried filters are washed three times for 5 minutes with 300 mM of NaCl/25 mM of trisodium citrate and once with 95 % ethanol and again air-dried. Evaluation is effected in a Matrix Packard 96-well counter (Packard, Downers, Grove, IL, USA). The RT activity is a measure of the reproduction of HIV-1. The compounds of formula I according to the invention inhibit virus reproduction when administered in the micromolar range, for example during 18 days after infection practically no increase in RT activity can be determined in the presence of preferably 5 to 50 μM concentrations of an inhibitor of the present invention (for example 0 to 100 counts per minute), while in the control high increase of RT activity can be found (for example more than 2000 counts per minute on day 18).
It can be shown that inhibition by a compound of formula I is mainly due to their ability to permeate into cells, a fact that can be demonstrated by a Fusion Induced Gene Stimulation Assay (FIGS-assay) that excludes cell surface effects as the underlying mechanism of action.
The compounds of the present invention can thus be used in the treatment of retroviral infections in warm-blooded animals, especially HIV, such as HTV-l, infections, and more specifically for the treatment of AIDS in humans. Also treatment of infected cells, e.g. lymphocytes, outside the body is possible in order to reintroduce healthy cells by transplantantion or injection, for example in order to improve the lymphocyte titer in patients with advanced AIDS.
The compounds of the present invention can also be used in the treatment of commercially valuable cell, such as lymphocyte, cultures against retroviral infections, especially against HIV, such as HTV-l, infections.
Preferred is a compound of formula I wherein
k is 0 to 20, preferably 0 to 12, most preferably 0 or 12, m is 3 to 10, preferably 4 to 6, most preferably 5, n is 1 to 10, preferably 1 to 4;
Ri is
hydrogen (especially preferred),
lower alkanoyl (especially preferred), such as acetyl, or further
substituted lower alkanoyl wherein the substituents are independently selected from - cycloalkyi with from 3 to 7 carbon atoms; - aryl selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, each of which is unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl,lower alkanoyl, sulfo, lower alkane- sulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxy¬ phosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
- carbamoyl;
- carbamoyl substituted at the nitrogen atom by one or two radicals selected from lower alkyl, carboxy-lower alkyl or lower alkoxy- carbonyl-lower alkyl;
- heterocyclyl selected from thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, chromanyl, cyclohexa[b]pyrrolyl, cyclohexa- [bjpyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S-dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by one or more substituents selected from lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxy¬ carbonyl, halogen, lower alkanoyl, nitro, oxo and cyano;
- hydroxy;
- lower alkoxy;
- lower alkanoyloxy;
- up to 3 halogen atoms and
- carboxy,
lower alkoxycarbonyl, 2-halo-lower alkoxycarbonyl,
aryl-lower alkoxycarbonyl wherein aryl is phenyl, 1- or 2-naphthyl, fluorenyl or phenyl mono- or poly-substituted by lower alkyl, phenyl, hydroxy, lower alkoxy, halogen or nitro,
heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, - pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, iso- quinolyl, 3,1-benzofuranyl, cyclohexa[b]pynolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyrrolidinyl, pyrrolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S- dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxycarbonyl, halogen, lower alkanoyl, nitro or cyano,
lower alkenyloxycarbonyl wherein the lower alkenyl radical is bonded to the bonding oxygen atom via a saturated carbon atom,
lower alkoxy-lower alkoxycarbonyl,
(lower alkoxy-lower alkoxy)-lower alkoxycarbonyl, such as 2-(2-methoxy- ethoxy )ethoxy carbonyl .
aryl-lower alkyl with up to three aryl groups, wherein each aryl is selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl which are unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di- lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, lower alkanoyl, sulfo, lower alkanesulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxyphosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
arylthio or aryl-lower alkylthio, wherein aryl is phenyl that is unsubstituted or substituted by lower alkyl, lower alkoxy, halogen or nitro,
1-lower alkanoyl-lower alk-l-en-2-yl,
lower alkoxycarbonyl-lower alk-l-en-2-yl,
tri-lower alkylsilyl,
arylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups,
(mono-, di- or tri-aryl)-lower alkylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups, or
2,2,5,7,8-pentamethylchroman-6-sulfonyl,
hydroxy,
a lower alkoxy group that is branched in the 1-position,
arylmethoxy having one or two aryl radicals, wherein aryl is phenyl that is unsubstituted or mono-, di- or tri-substituted by lower alkyl, lower alkoxy, hydroxy, halogen or nitro,
1-lower alkoxy-lower alkoxy, 1 -lower alkylthio-lower alkoxy, benzoylmethoxy wherein benzoyl is unsubstituted or substituted by halogen,
2-halo-lower alkoxy,
2-(tri-substituted silyl)-lower alkoxy wherein the substituents are each independently of the others selected from lower alkyl, phenyl-lower alkyl, C3-C7-cycloalkyl or phenyl,
tri-lower alkylsilyloxy,
amino, or
a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyrrolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, moφholinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyrrolidinyl an 4-moφholinyl;
amino R2 being most preferred;
any A being present and any B being present are independently selected from the group comprising a bivalent radical, bonded via its α-amino and its α-carbonyl group, of an amino acid selected from glycine alanine, valine, norvaline, leucine, isoleucine, norleucine, α-amino-n-decanoic acid, serine, homoserine, threonine, methionine, cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine, tyrosine, 4-amino- phenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenyl- alanine, β-phenylserine,phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan, indoline-2-carboxylic acid, 1,2,3,4-tetrahydroiso- quinoline-3-carboxylic acid, aspartic acid, asparagine, aminomalonic acid, amino¬ malonic acid onoamide, glutamic acid, glutamine, histidine, arginine, lysine, Ne-benzyl-Ne-methyl-lysine, Ne,Ne-dibenzyl-lysine, δ-hydroxylysine, ornithine, α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyrrolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino-γ-(4-moφholinyl), α-amino-γ-(3-indolyl)-butyric acid, α,γ-diaminobutyric acid and α,β-diamino- propionic acid; especially preferably the bivalent radical of an α-amino acid, bonded via its α-amino and its α-carbonyl group, selected from glycine, alanine, valine, leucine, isoleucine, α-amino-decanoic acid, serine, threonine, proline, phenylalanine, tyrosine, 4-nitrophenylalanine, glutamic acid, arginine, lysine, Ne-benzyl-N6-methyl-lysine, Ne,N6-dibenzyl-lysine, α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyrrolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino-γ-(4-moφholinyl)-butyric acid and α-amino-γ-(3-indolyl)-butyric acid;
it being possible for each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form, preferably in the L-form (more preferred in the case of the radical A) or in the D-form (more preferred in the case of the radical B);
most preferably the moiety A being a bivalent radical of an α-amino acid selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present preferably in the L-form (where an asymmetric α-carbon atom is present), the moiety B being a bivalent radical of an α-amino acid selected from lysine, arginine and proline, each of which is present preferably in the D-form;
and each X being present is a bivalent radical of the partial formula II,
Figure imgf000027_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
- amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl,
- N-lower alkylamino-, such as N-methylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, N,N-di(phenyl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or guanidino-lower alkyl, such as 3-guanidinopropyl (lower alkyl preferably being methyl, ethyl or propyl),
- pyridyl-lower alkyl, for example (2-pyridyl)-lower alkyl, such as 2-(2-ρyridyl)ethyl,
- pyrrolidinyl-lower alkyl, for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pynolidinyl)ethyl,
- moφholinyl-lower alkyl, for example (4-moφholinyl)-lower alkyl, such as 2-(4-moφholinyl)ethyl,
or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus (that is the underlined moiety in the following representation of the bivalent radical -(X)m-, namely -X-X-X-X- ...), may in addition be a side chain selected from
- aryl-lower alkyl wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
- carboxy-lower alkyl, such as 2-carboxyethyl,
- carbamoyl-lower alkyl (H2N-C(=O)-lower alkyl), such as 2-carbamoylethyl or 3-carbamoylpropyl,
- furyl-lower alkyl, for example (2-furyl)-lower alkyl, such as (2-furyl)methyl, and
- indolyl-lower alkyl, for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl;
or (alternatively to the above-mentioned residues or included in their group) selected from (basic) (acridin-9-yl)amino-lower alkyl, (3,6-bis(di- methylamino)acridin-9-yl)amino-lower alkyl or (6-chloro-9-methoxyacri- din-9-yl)amino-lower alkyl, such as 2-(acridin-9-yl)amino-ethyl, 2-(3,6-bis(dimethylamino)acridin-9-yl)amino-ethyl or 2-(6-chloro-9- methoxyacridin-9-yl)amino-ethyl, (basic) (amino)-(phenyl)-lower alkyl, such as 2-amino-2-phenylethyl, (basic) (amino-loweralkylphenoxy)-lower alkyl, such as 2-(4-(2-aminoethyl)phenoxy)-ethyl, and (basic) oxo- pyrrolidinyl-lower alkyl, such as 3-(2-oxopyrrolidin-l-yl)ρropyl; the bivalent radicals X together preferably forming a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than lower alkyl or benzyl;
and
R4 and R5 each represent hydrogen;
or a salt thereof.
More prefened is a compound of formula I, wherein
k is 0 to 12, more preferably 0 or 12, most preferably 0, m is 4 to 6, most preferably 5, n is 1 to 4;
Rj is
hydrogen or
lower alkanoyl, such as acetyl,
R2 is amino, or further
a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyrrolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, moφholinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyrrolidinyl an 4-moφholinyl;
amino R being most preferred; any A being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or preferably in the L-form (where an asymmetric α-carbon atom is present);
any B being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
and any X being present is a bivalent radical of the partial formula II,
Figure imgf000030_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
- amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl,
- N-lower alkylamino-, such as N-methylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, such as N-benzyl-N-methylamino-, N,N-di(phenyl-lower alkyl)-amino-, such as N,N-dibenzylamino-, or guanidino-lower alkyl, such as 3-guanidinopropyl (lower alkyl preferably being methyl, ethyl or propyl),
- pyridyl-lower alkyl, for example (2-pyridyl)-lower alkyl, such as 2-(2-pyridyl)ethyl,
- pyrrolidinyl-lower alkyl, for example (l-pyrrolidinyl)-lower alkyl, such as 2-(l-pyrrolidinyl)ethyl, and
- moφholinyl-lower alkyl, for example (4-moφholinyl)-lower alkyl, such as 2-(4-moφholinyl)ethyl, or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus (that is the underlined moiety in the following representation of the bivalent radical -(X)m-, namely -X-X-X-X- ...), may in addition be a side chain selected from
- aryl-lower alkyl wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
- carboxy-lower alkyl, such as 2-carboxyethyl,
- carbamoyl-lower alkyl (H2N-C(=O)-lower alkyl), such as 2-carbamoylethyl or 3-carbamoylpropyl,
- furyl-lower alkyl, for example (2-furyl)-lower alkyl, such as (2-furyl)methyl, and
- indolyl-lower alkyl, for example (3-indolyl)-lower alkyl, such as 2-(3-indolyl)-ethyl;
or (alternatively to the above-mentioned residues or included in their group) selected from (basic) (acridin-9-yl)amino-lower alkyl, (3,6-bis(di- mefhylamino)acridin-9-yl)amino-lower alkyl or (6-chloro-9-methoxyacri- din-9-yl)amino-lower alkyl, such as 2-(acridin-9-yl)amino-ethyl, 2-(3,6-bis(dimethylamino)acridin-9-yl)amino-ethyl or 2-(6-chloro-9- methoxyacridin-9-yl)amino-ethyl, (basic) (amino)-(phenyl)-lower alkyl, such as 2-amino-2-phenylethyl, (basic) (amino-loweralkylphenoxy)-lower alkyl, such as 2-(4-(2-aminoethyl)phenoxy)-ethyl, and (basic) oxo- pynolidinyl-lower alkyl, such as 3-(2-oxopyrrolidin-l-yl)propyl;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than lower alkyl or benzyl;
most preferably R3 being
- amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl, or
- guanidino-lower alkyl, such as 3-guanidinopropyl
(lower alkyl preferably being methyl, ethyl or propyl)
or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus (that is the underlined moiety in the following representation of the bivalent radical -(X)m-, namely -X-X-X-X- ...), may in addition be an - aryl-lower alkyl side chain wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, aryl¬ methoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro; phenyl-lower alkyl, especially benzyl, being most preferred in that position;
and R4 and R5 each represent hydrogen;
or a salt thereof.
Still more prefeπed is a compound of formula I wherein
k is 0 or 12, most preferably 0, m is 4 to 6, most preferably 5, n is 1 to 4;
R, is
hydrogen or,
lower alkanoyl, such as acetyl,
R, is ammo,
any A being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or preferably in the L-form (where an asymmetric α-carbon atom is present);
any B being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
and any X being present is a bivalent radical of the partial formula II,
Figure imgf000033_0001
wherein
R3 represents a side chain selected from the group comprising
- amino-lower alkyl, such as 4-aminobutyl or 6-aminohexyl, and
- guanidino-lower alkyl, such as 3-guanidinopropyl, or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus (that is the underlined moiety in the following representation of the bivalent radical -(X)m-, namely -X-X-X-X- ...), may in addition be a phenyl-lower alkyl side chain, such as benzyl;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than benzyl;
and R and R5 each represent hydrogen;
or a salt thereof.
Most preferred is a compound mentioned in the examples, or a (preferably pharmaceutically acceptable) salt thereof.
The compounds of the present invention can be synthesized according to known procedures, especially by a process comprising
a) reacting a fragment of a compound of formula I, which has a free carboxy group, or a reactive derivative thereof, with a complementary fragment that has an amino group with at least one free hydrogen atom, or with a reactive derivative thereof, with formation of an amide bond; in the mentioned fragments free functional groups with the exception of those that participate in the reaction if required being present in protected form; and removing any protecting groups present; or
b) for the synthesis of compounds of formula I wherein k = 0, Rj is hydrogen and the N-terminal X is a bivalent radical of the partial formula II,
Figure imgf000034_0001
wherein R3 is a residue from the group comprising lower alkyl or substituted lower alkyl (preferably as defined above for R6 in formula III), while R2, R4, R5, A, B, X, m and n have the meanings given for compounds of the formula I,
alkylating an amino compound of the formula IV,
H2N-R3 (IV),
wherein R3 has the meanings given above, with a compound of the formula V
Z-C(R4R5)-C(=O)-(X)m.r(B)n-R2' (V)
wherein Z is a nucleofugal leaving group, R2' has the same meaning as R2 in compounds of formula I or is a resin for solid phase synthesis and R4, R5, X, B, m and n have the meanings given for compounds of formula I, under nucleophilic substitution, in the mentioned starting materials free functional groups with the exception of those that participate in the reaction if required being present in protected form; and removing any protecting groups and cleaving from any resin for solid phase synthesis being present;
and, if desired, transforming a salt of an obtainable compound of formula I into the free compound or a different salt or an obtainable free compound of formula I into a salt, and/or separating obtainable mixtures of isomers of compounds of formula I into the individual isomers.
In the following, more detailed description of the preferred process conditions, Rj, R2, R3, R4, R5, A, B, X, k, m and n have the meanings given for compounds of the formula I, if not mentioned otherwise.
Process a):
The compounds of the present invention preferably can be readily prepared according to well-established, standard liquid or. preferably, solid-phase peptide synthesis methods, general descriptions of which are broadly availabe, for example in J.M. Stewart and J.D. Young, Solid Phase Peptide Synthesis, 2nd edition, Pierce Chemical Company, Rockford, Illinois (1984), in M. Bodanzsky and A. Bodanzsky, The Practice of Peptide Synthesis, Springer Verlag, New York (1984); and Applied Biosystems 430A Users Manual, ABI Inc., Foster City, California, but may also be prepared in solution or by a combination of solid-phase and solution chemistry.
A fragment with a free carboxy group can be an amino acid (if required, in suitably protected form) or a di-, tri- or other oligopeptide (the term "peptide" here also comprising peptoid structures) or also, e.g. in the case of the synthesis of derivatives of formula I with acylated terminal amino group, the acylating carbonic acid, especially acetic acid. A fragment that has an amino group with at least one free hydrogen atom (= a group -NH-) can also be a single amino acid, a di-, tri- or oligopeptide or, in the case of preparation of peptamides, ammonia or mono- or disubstituted ammonia.
Reactive derivatives of carbonic acids are preferably reactive esters or reactive anhydrides, or reactive cyclic amides. Reactive carbonic acid derivatives can also be formed in situ.
A reactive derivative of an "amino group with at least one free hydrogen" is preferably derivatized by the reaction with a phosphite, such as diethyl-chlorophosphite, 1,2-phenylene-chlorophosphite, ethyl-dichlorophosphite, ethylene-chlorophosphite or tetraethyl-pyrophosphite; or is present in the form of a carbamic acid chloride wherein the amino group participating in the reaction is subtituted by halocarbonyl, such as chlorocarbonyl. The reaction steps required e.g. for the synthesis of amide bonds usually depend on the type of activation of the carboxylic group participating in the reaction. The reactions normally run in the presence of a condensing agent or, when activating the carboxylic acids in the form of anhydrides, of an agent that binds the carboxylic acid formed. In some cases it is also possible to add chaotropic agents such as LiF in NB-methylpynolidone. The reactions are especially carried out in a temperature range from -30 to +150 °C, preferably from +10 to +70 °C, and, most preferably, from +20 to +50 °C, if appropriate, in an inert gas atmosphere, e.g. under nitrogen or argon.
If desired or appropriate, unreacted amino groups can be acylated after a reaction cycle, e.g. by acetylation of unreacted amino groups with 10 ml acetic anhydride/pyridine/DMA (1:1:8), thus facilitating later purification of the final product.
In general, a suitably protected amino acid [an α-amino acid or (for the introduction of peptoid structures) an amino acid of formula lib
Figure imgf000036_0001
wherein R3, R4 and R5 have the meaning given under formula II above, R3 not being hydrogen] as a ligand is attached via its carboxyl group (-COOH) to a derivatized, insoluble polymeric support, e.g. a cross-linked styrene or polyamide resin, such as a 4-(2',4'-dimethoxyphenyl-[hydroxy- or amino-]methyl)-phenyoxymethyl-polystyrene resin by a condensation reaction. "Suitably protected" refers to the presence of protecting groups on the α-amino group (α-NH2 or α-NHR3) and any side-chain functional group (if present) of the amino acid. Di-, tri- or other oligopeptides can be used instead of the amino acids as building blocks (fragments).
Synthesis proceeds in a stepwise, cyclical fashion by successively removing the α-NH or α-NHR3 protecting group and then coupling an activated fragment (e.g. an amino acid, di-, tri- or oligopeptide) to the deprotected α-NH2 or α-NHR3. Preferably, activation of the α-COOH group of the amino acid to be attached by the condensation reaction is effected (i) directly with a carbodiimide, e.g. dicyclohexylcarbodiimide (DCC), N-ethyl-N'-(3-di- methylaminopropyl)-carbodiimide, N,N'-diethylcarbodiimide or N,N'-diisopropylcarbo- diimide (DICD); with a carbonyl compound such as carbonyldiimidazole; with 1,2-ox- azolium compounds such as 2-ethyl-5-phenyl-l,2-oxazolium-3 '-sulfonate and 2-tert- butyl-5-methylisoxazolium perchlorate; with acylamino compounds such as 2-ethoxy- l-ethoxycarbonyl-l,2-dihydroquinoline; with an uronium compound such as 2-(lH-benzo- triazol-l-yl)-l,l,3,3-tetramethyluronium tetrafluoroborate (TBTU) or 2-(pyridon- l-yl)-l,l,3,3-tetramethyluronium tetrafluoroborate = O-(l,2-dihydro-2-oxo-l-pyridyl)- -N,N,N',N'-tetramethyluronium-tetrafluoroborate (TPTU); or phosphonium compounds such as benzotriazol-l-yl-oxy-tris(dimethylamino)-phosphonium hexafluorophosphate (BOP) or benzotriazol-1-yl-oxy-pyrrolidino-phosphonium hexafluorophosphate (PyBOP);
(ii) via formation of the symmetric anhydride (obtainable, for example, by condensation of the corresponding acid in the presence of a carbodiimide or 1-diethylaminopropyne; symmetric anhydrides method), and/or
(iii) by formation of an "active ester", e.g. a hydroxybenzotriazole (HOBT), pentafluorophenyl, 4-nitrophenyl or N-hydroxysuccinimide ester.
Useful acid binding agents that can be employed in the condensation reactions are, for example, alkaline metals, carbonates or bicarbonates, such as sodium or potassium carbonate or bicarbonate (if appropriate, together with a sulfate), or organic bases such as sterically hindered organic nitrogen bases, for example tri-lower alkylamines, such as N,N-diisopropyl-N-ethylamine,
Reactive groups in the monomers of ligands or in the resin-bound or free intermediates resulting from one or more coupling steps can be protected by third groups as protecting groups that are customarily used in peptide synthesis. Examples of protecting groups, their introduction and their removal are, for example, described in standard works such as "Protective groups in Organic Chemistry", Plenum Press, London, New York 1973; "Methoden der organischen Chemie", Houben-Weyl, 4. edition, Vol. 15/1, Georg-Thieme Verlag, Stuttgart 1974; Th. W. Greene, "Protective Groups in Organic Synthesis", John Wiley & Sons, New York 1981; Atherton et al., "Solid Phase Peptide Synthesis - A Practical Approach", IRL Press Oxford University, 1984; Jones, "The Chemical Synthesis of Peptides", Oxford Science Publications, Clavendon Press Oxford, 1991; and Bodanszky, "Peptide Chemistry", Springer Verlag Berlin, 1988. The term "protecting groups" comprises also resins used for solid phase synthesis, preferably those specifically mentioned above and below.
Examples for hydroxy protecting groups are acyl radicals, such as tert-lower alkoxycarbonyl radicals, for example tert-butoxycarbonyl, etherifying groups, such as tert-lower alkyl groups, for example t-butyl, or silyl- or tin radicals, such as tert-butyl-dimethylsilyl or the tri-n-butyltin radical.
Carboxy groups can be protected by groups as defined above for the C-terminal protecting groups R2, preferably by esterifying groups selected from those of the tert-butyl type, from benzyl, from trimethylsilylethyl and from 2-triphenylsilyl groups .
Amino or guanidino groups can be protected by removable acyl groups or by arylmethyl, etherified mercapto, 2-acyl-lower alk-1-enyl, a silyl group or an organic sulfonyl group (preferably as defined above for an "amino substituent other than acyl" Rj) or tin amino protecting groups; tert-butoxycarbonyl, allyloxycarbonyl, benzyloxycarbonyl, 4-nitro- benzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2-bromobenzyloxycarbonyl (especially the tyrosine OH group), diphenylmethoxycarbonyl, nitrophenylsulfenyl, 2,2,2-trichloro- ethoxycarbonyl, 2,2,5,7,8-pentamefhylchroman-6-sulfonyl (PMC), 2,2,4,6,7-pentamethyl- dihydrobenzofuran-5-sulfonyl (Pbf) or 4-methoxy-2,3,6-trimethylbenzenesulfonyl (Mtr) being especially preferred.
Imino groups (e.g. in imidazole) can be protected by 2,4-dinitrophenyl or p-toluene- sulfonyl, or (e.g. in indole) by formyl.
Mercapto groups can be protected, e.g., by acetamidomethyl, by trityl or by p-methylbenzyl.
These protective groups are usually removed after the complete synthesis of the resin-bound molecule by conventional methods of peptide chemistry, conveniently by treatment with 95 % trifluoroacetic acid. In some cases, strong nucleophiles, such as 1,2-ethanedithiol, may be additionally added to capture the generated compounds resulting from the protecting groups.
Further methods of removing protective groups are known in the art and comprise, inter alia, β-elimination, solvolysis, hydrolysis, alcoholysis, acidolysis, photolysis, treatment with a base or reduction.
Illustrative examples of the combination of protective group types are:
α-amino group of ligand additional functional group not to be reacted (in side chains)
Fmoc type acid-labile or allyl type trityl type acid-labile or allyl type allyloxycarbonyl acid-labile
Boc or nitrophenylsulfenyl allyl type type allyloxycarbonyl type β-elimination type trityl type β-elimination type
Groups of the β-elimination type are typically protective groups of the fluorenylmethyl type.
The two prefeπed methods of solid phase peptide synthesis are the Boc and the Fmoc methods, which are named with reference to their use of the tert-butoxycarbonyl (Boc) or 9-fluorenylmethyloxycarbonyl (Fmoc) group, respectively, to protect the α-NH2 or α-NHR3 of the amino acid residue to be coupled.
In the more established Boc method, the acid-lability of the Boc group is exploited and trifluoroacetic acid (TFA) treatment is used in order to remove the protective group. Prefeπed third groups as protecting groups (for functional groups in side chains) are relatively stable in weak acid, e.g. TFA. Most can be cleaved by strong acids such as hydrofluric acid (HF) or trifluoromethanesulfonic acid. A small number of side chain groups (e.g. 2,4-dinitrophenyl protected imino in the histidyl side chain, may require a separate deprotection step, e.g. treatment with thiophenol or ammonolysis. After synthesis, the product is typically cleaved from the resin and simultaneously deprotected by HF treatment at low temperature (e.g. around 0 °C).
The Fmoc-group can be cleaved off preferably in the presence of a mild nitrogen base, preferably piperidine, in an inert solvent, preferably dimethyl acetamide, thereby allowing the use of side-chain protecting groups which are labile to milder treatment, e.g. TFA.
An acid labile ether resin such as HMP-resin (p-hydroxymethylphenoxymethyl poly¬ styrene), 4-(2',4'-dimethoxyphenyl-hydroxymethyl)-phenoxymethyl-polystyrene or preferably a resin with a benzyloxy- or alkyloxy linker (see Wang, J. Amer. Chem. Soc. 95, 1328 (1973); or, for the synthesis of compounds with a C-terminal amino group R2 (in amide bond) which are prefeπed, 4-(2',4'-dimethoxyphenyl-aminomethyl)-phenoxy- methyl-polystyrene (Rink et al., Tetrahedr. Lett. 28(33), 3787-90 (1987) is used as the solid support, permitting simultaneous cleavage/deprotection in TFA.
Process b):
In the starting materials of formula V, Z is preferably a nucleofugal group, preferably aryl- sulfonyloxy, such as toluenesulfonyloxy, lower alkanesulfonyloxy, such as methane- sulfonyloxy, or especially halogen, such as chlorine, bromine or iodine, more especially chlorine or iodine and most especially bromine.
The reaction is caπied out under conditions customary for a nucleophilic substitution, pre¬ ferably using aprotic solvents, such as ketones, for example a di-lower alkyl ketone, such as acetone, nitriles, for example a lower alkylnitrile, such as acetonitrile, carboxylic acid amides, for example a di-lower alkyl-lower alkanoylamide, such as dimethylformamide or dimethylacetamide, di-lower alkyl sulfoxides, such as dimethyl sulfoxide, hexamethyl- phosphoric acid triamide or ethers, such as di-lower alkyl ethers, for example diethyl ether, or cyclic ethers, such as tetrahydrofuran or dioxane, or also in protic solvents, such as alcohols, especially lower alkanols, for example methanol or ethanol, or mixtures of two or more of the mentioned solvents, the prefeπed temperature range being from -10 °C up to 70 °C, preferably in the range from 10 to 40 °C, for example at room temperature.
The protecting groups, their introduction, their removal, the resins used for solid phase synthesis and methods of cleavage from them are preferably analogous to those described under process b).
Additional process steps
Salts of compounds of formula I having at least one salt-forming group may be prepared in a manner known er se. For example, salts of compounds of formula I having acid groups may be formed, for example, by treating the compounds with metal compounds, such as alkali metal salts of suitable organic carboxylic acids, e.g. the sodium salt of 2-ethyl- hexanoic acid, with organic alkali metal or alkaline earth metal compounds, such as the coπesponding hydroxides, carbonates or hydrogen carbonates, such as sodium or potassium hydroxide, carbonate or hydrogen carbonate, with coπesponding calcium compounds or with ammonia or a suitable organic amine, stoichiometric amounts or only a small excess of the salt-forming agent preferably being used. Acid addition salts of compounds of formula I are obtained in customary manner, e.g. by treating the compounds with an acid or a suitable anion exchange reagent. Internal salts of compounds of formula I containing acid and basic salt-forming groups, e.g. a free carboxy group and a free amino group, may be formed, e.g. by the neutralisation of salts, such as acid addition salts, to the isoelectric point, e.g. with weak bases, or by treatment with ion exchangers.
Salts can be converted in customary manner into the free compounds; metal and ammonium salts can be converted, for example, by treatment with suitable acids, and acid addition salts, for example, by treatment with a suitable basic agent.
Mixtures of isomers obtainable according to the invention can be separated in a manner known per se into the individual isomers; diastereoisomers can be separated, for example, by partitioning between polyphasic solvent mixtures, recrystallisation and/or chromato¬ graphic separation, for example over silica gel, and racemates can be separated, for example, by the formation of salts with optically pure salt-forming reagents and separation of the mixture of diastereoisomers so obtainable, for example by means of fractional crystallisation, or by chromatography over optically active column materials.
Starting materials:
The present invention relates also to novel starting materials and/or intermediates and to processes for their preparation. The starting materials used and the reaction conditions selected are preferably those that result in the compounds described as being prefeπed.
Unless a specific method of synthesis is indicated for starting materials, the starting materials are known, can be prepared according to processes known per se and/or are available commercially.
For example, suitably protected and/or preactivated D-, D,L- or L- amino acids, di-, tri- or oligopeptides, derivatized and/or preloaded resins, the ancillary reagents and solvents required for either Boc or Fmoc peptide synthesis are commercially available from various suppliers or can be prepared readily according to standard procedures. In addition, and di-, tri- or oligopeptoids can be prepared readily according to standard procedures. In addition, automated peptide synthesizers with optimized, preprogrammed Boc and Fmoc synthesis cycles are available from numerous sources.
The compounds of formula V can be prepared by condensation of a carbonic acid of the formula (VI),
Z-C(R4R5)-COOH (VI)
or a reactive derivative thereof, wherein Z has the meanings given above for compounds of formula V and R4 and R5 have the meanings given for compounds of formula I, with an amino compound of the formula
H-(X)m.r(B)n-R2' (V)
wherein R2' has the same meaning as R2 in compounds of formula I or is a resin for solid phase synthesis, while X, B, m and n have the meanings given in the definition of compounds of formula I. If necessary, functional groups that are not to be reacted are present in protected form. The condensation conditions, protective groups, their introduction etc. are analogous to those described for the synthesis of compounds of formula I under process a).
An importatn starting material is also the compound of the formula
H-Nahg-Narg-Narg-Nphe-Narg-NH2 (SEQ ID NO:2) which is very active in the above-mentioned indications, or a salt thereof.
General process conditions
The following applies in general to all processes mentioned hereinbefore and hereinafter:
Functional groups in starting materials the reaction of which is to be avoided, especially carboxy, amino, hydroxy, mercapto and sulfo groups, can be protected by suitable protect¬ ing groups (conventional protecting groups) which are customarily used in the synthesis of peptide compounds, and also in the synthesis of cephalosporins and penicillins as well as nucleic acid derivatives and sugars. These protecting groups may already be present in the precursors and are intended to protect the functional groups in question against undesired secondary reactions, such as acylation, etherification, esterification, oxidation, solvolysis, etc.. In certain cases the protecting groups can additionally cause the reactions to proceed selectively, for example stereoselectively. It is characteristic of protecting groups that they can be removed easily, i.e. without undesired secondary reactions taking place, for example by solvolysis, reduction, photolysis, and also enzymatically, for example also under physiological conditions, and, especially, that they are not present in the end products.
The protection of functional groups by such protecting groups, the protecting groups them¬ selves and the reactions for their removal are described, for example, in standard works such as J. F. W. McOmie, "Protective Groups in Organic Chemistry", Plenum Press, London and New York 1973, in Th. W. Greene, "Protective Groups in Organic Synthesis", Wiley, New York 1981, in "The Peptides", Volume 3 (E. Gross and J. Meienhofer, eds.), Academic Press, London and New York 1981, in "Methoden der organischen Chemie", Houben-Weyl, 4th edition, Volume 15/1, Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, "Aminosauren, Peptide, Proteine" ("Amino acids, peptides, proteins"), Verlag Chemie, Weinheim, Deerfield Beach and Basle 1982, and in Jochen Lehmann, "Chemie der Kohlenhydrate: Monosaccharide und Derivate" ("The Chemistry of Carbohydrates: monosaccharides and derivatives"), Georg Thieme Verlag, Stuttgart 1974.
When several protected functional groups are present, if desired the protecting groups can be so selected that more than one such group can be removed simultaneously, for example by acidolysis, such as by treatment with trifluoroacetic acid, or with hydrogen and a hydrogenation catalyst, such as a palladium-on-carbon catalyst. Conversely, the groups can also be so selected that they cannot all be removed simultaneously, but rather in a desired sequence, the coπesponding inteπnediates being obtained.
In view of the close relationship between the compounds of formula I and their salts and starting materials (starting materials and intermediates) in free form and in the form of their salts, any reference hereinbefore and hereinafter to a free compound or a salt thereof is to be understood as meaning also the corresponding salt or free compound, respectively, where appropriate and expedient. All the above-mentioned process steps can be carried out under reaction conditions that are known per se, preferably those mentioned specifically, in the absence or, customarily, in the presence of solvents or diluents, preferably solvents or diluents that are inert towards the reagents used and are solvents therefor, in the absence or presence of catalysts, condensation agents or neutralising agents, for example ion exchangers, such as cation exchangers, e.g. in the H+ form, depending on the nature of the reaction and/or of the reactants at reduced, normal or elevated temperature, for example in a temperature range of from approximately -100°C to approximately 190°C, preferably from approxi¬ mately -80°C to approximately 150°C, for example at from -80 to -60°C, at room temperature, at from -20 to 40°C or at reflux temperature, under atmospheric pressure or in a closed vessel, where appropriate under pressure, and/or in an inert atmosphere, for example under an argon or nitrogen atmosphere.
At all stages of the reactions, mixtures of isomers that are formed can be separated into the individual isomers, for example diastereoisomers or enantiomers, or into any desired mix¬ tures of isomers, for example racemates or mixtures of diastereoisomers, for example ana¬ logously to the methods described under "Additional process steps".
The solvents from which those solvents that are suitable for any particular reaction may be selected include, for example, water, esters, such as lower alkyl-lower alkanoates, for example ethyl acetate, ethers, such as aliphatic ethers, for example diethyl ether, or cyclic ethers, for example tetrahydrofuran, liquid aromatic hydrocarbons, such as benzene or toluene, alcohols, such as methanol, ethanol or 1- or 2-propanol, nitriles, such as aceto¬ nitrile, halogenated hydrocarbons, such as methylene chloride, acid amides, such as dimethylformamide, bases, such as heterocyclic nitrogen bases, for example pyridine, carboxylic acid anhydrides, such as lower alkanoic acid anhydrides, for example acetic anhydride, cyclic, linear or branched hydrocarbons, such as cyclohexane, hexane or iso- pentane, or mixtures of those solvents, for example aqueous solutions, unless otherwise indicated in the description of the processes. Such solvent mixtures may also be used in working up, for example by chromatography or partitioning.
The compounds, including their salts, may also be obtained in the form of hydrates, or their crystals may, for example, include the solvent used for crystallisation.
If necessary, protected starting materials may be used in all process steps and the protecting groups may be removed at suitable stages of the reaction.
The invention relates also to those forms of the process in which a compound obtainable as intermediate at any stage of the process is used as starting material and the remaining process steps are caπied out, or in which a starting material is formed under the reaction conditions or is used in the form of a derivative, for example in protected form or in the form of a salt, or a compound obtainable by the process according to the invention is produced under the process conditions and processed further in situ. In the process of the present invention there are preferably used those starting materials which result in t e* compounds of formula I described at the beginning as being especially valuable. Special preference is given to reaction conditions that are analogous to those mentioned in the Examples.
Pharmaceutical Compositions:
The invention relates also to pharmaceutical compositions comprising compounds of formula I.
The pharmacologically acceptable compounds of the present invention may be used, for example, for the preparation of pharmaceutical compositions that comprise an effective amount of the active ingredient together or in admixture with a significant amount of inorganic or organic, solid or liquid, pharmaceutically acceptable carriers.
The invention relates also to a pharmaceutical composition that is suitable for administra¬ tion to a warm-blooded animal, especially a human (or to cells or cell lines derived from a warm-blooded animal, especially a human, e.g. lymphocytes), for the treatment or prevention of (= prophylaxis against) a disease that responds to inhibition of retroviral Tat/TAR interaction retroviral protease; especially inhibition of HIV, more preferably HIV-1, Tat/TAR interaction; for example a retroviral infection such as HIV infection, more especially AIDS, comprising an amount of a compound of formula I or a pharmaceutically acceptable salt thereof, effective for the inhibition of the Tat/TAR interaction, especially for the treatment of HIV infection, together with at least one pharmaceutically acceptable caπier.
The pharmaceutical compositions according to the invention are those for enteral, such as nasal, rectal or oral, or parenteral, such as intramuscular or intravenous, administration to warm-blooded animals (humans and animals), that comprise an effective dose of the pharmacological active ingredient, alone or together with a significant amount of a pharmaceutically acceptable carrier. The dose of the active ingredient depends on the species of warm-blooded animal, the body weight, the age and the individual condition, individual pharmacokinetic data, the disease to be treated and the mode of administration.
The invention relates also to a method of treating diseases caused by viruses, especially by retroviral infections, for example HIV infection, including AIDS, which comprises administering a prophylactically or especially therapeutically effective amount of a compound of formula I according to the invention, especially to a warm-blooded animal, for example a human, who on account of one of the mentioned diseases, especially HTV-infection, including AIDS, requires such treatment. The dose to be administered to warm-blooded animals, for example humans of approximately 70 kg body weight, is from approximately 3 mg to approximately 3 g, preferably from approximately 10 mg to approximately 1.5 g, for example approximately from 100 mg to 1000 mg per person per day, divided preferably into 1 to 3 single doses which may, for example, be of the same size. Usually, children receive half of the adult dose.
The pharmaceutical compositions comprise from approximately 1 % to approximately 95 %, preferably from approximately 20 % to approximately 90 %, active ingredient. Pharmaceutical compositions according to the invention may be, for example, in unit dose form, such as in the form of ampoules, vials, suppositories, dragees, tablets or capsules.
The pharmaceutical compositions of the present invention are prepared in a manner known per se, for example by means of conventional dissolving, lyophilising, mixing, granulating or confectioning processes.
Solutions of the active ingredient, and also suspensions, and especially isotonic aqueous solutions or suspensions, are preferably used, it being possible, for example in the case of lyophilised compositions that comprise the active ingredient alone or together with a caπier, for example mannitol, for such solutions or suspensions to be produced prior to use. The pharmaceutical compositions may be sterilised and/or may comprise excipients, for example preservatives, stabilisers, wetting and/or emulsifying agents, solubilisers, salts for regulating the osmotic pressure and/or buffers, and are prepared in a manner known per se, for example by means of conventional dissolving or lyophilising processes. The said solutions or suspensions may comprise viscosity-increasing substances, such as sodium carboxymethylcellulose, carboxymethylcellulose, dextran, polyvinylpyπolidone or gelatin.
Suspensions in oil comprise as the oil component the vegetable, synthetic or semi- synthetic oils customary for injection puφoses. There may be mentioned as such especially liquid fatty acid esters that contain as the acid component a long-chained fatty acid having from 8 to 22, especially from 12 to 22, carbon atoms, for example lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, stearic acid, arachidic acid, behenic acid or coπesponding unsaturated acids, for example oleic acid, elaidic acid, erucic acid, brasidic acid or linoleic acid, if desired with the addition of anti¬ oxidants, for example vitamin E, β-carotene or 3,5-di-tert-butyl-4-hydroxytoluene. The alcohol component of those fatty acid esters has a maximum of 6 carbon atoms and is a mono- or poly-hydric, for example a mono-, di- or tri-hydric, alcohol, for example methanol, ethanol, propanol, butanol or pentanol or the isomers thereof, but especially glycol and glycerol. The following examples of fatty acid esters are therefore to be mentioned: ethyl oleate, isopropyl myristate, isopropyl palmitate, "Labrafil M 2375" (polyoxyethylene glycerol trioleate, Gattefosse, Paris), "Miglyol 812" (triglyceride of saturated fatty acids with a chain length of C8 to C]2, Hϋls AG, Germany), but especially vegetable oils, such as cottonseed oil, almond oil, olive oil, castor oil, sesame oil, soybean oil and more especially groundnut oil.
The injection compositions are prepared in customary manner under sterile conditions; the same applies also to introducing the compositions into ampoules or vials and sealing the containers.
Pharmaceutical compositions for oral administration can be obtained by combining the active ingredient with solid carriers, if desired granulating a resulting mixture, and processing the mixture, if desired or necessary, after the addition of appropriate excipients, into tablets, dragee cores or capsules. It is also possible for them to be incoφorated into plastics caπiers that allow the active ingredients to diffuse or be released in measured amounts.
Suitable carriers are especially fillers, such as sugars, for example lactose, saccharose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, and binders, such as starch pastes using for example corn, wheat, rice or potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or polyvinyl¬ pyπolidone, and/or, if desired, disintegrators, such as the above-mentioned starches, also carboxymethyl starch, crosslinked polyvinylpyπolidone, agar, alginic acid or a salt thereof, such as sodium alginate. Excipients are especially flow conditioners and lubricants, for example silicic acid, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol. Dragee cores are provided with suitable, optionally enteric, coatings, there being used, inter alia, concentrated sugar solutions which may comprise gum arabic, talc, polyvinylpyπolidone, polyethylene glycol and/or titanium dioxide, or coating solutions in suitable organic solvents, or, for the preparation of enteric coatings, solutions of suitable cellulose preparations, such as ethylcellulose phthalate or hydroxypropylmethylcellulose phthalate. Capsules are dry-filled capsules made of gelatin and soft sealed capsules made of gelatin and a plasticiser, such as glycerol or sorbitol. The dry-filled capsules may comprise the active ingredient in the form of granules, for example with fillers, such as lactose, binders, such as starches, and/or glidants, such as talc or magnesium stearate, and if desired with stabilisers. In soft capsules the active ingredient is preferably dissolved or suspended in suitable oily excipients, such as fatty oils, paraffin oil or liquid polyethylene glycols, it being possible also for stabilisers and/or antibacterial agents to be added. Dyes or pigments may be added to the tablets or dragee coatings or the capsule casings, for example for identification puφoses or to indicate different doses of active ingredient.
Examples
Embodiments of the invention are described in the following specific examples which are not to be construed to be intended to limit the scope of the invention in any way, but serve merely for illustration:
Temperatures, if not mentioned: room temperature/ambient temperature. In mixtures, relations of parts of solvent or eluent or reagent mixtures in liquid form are given as volume relations (v/v), if not indicated otherwise.
Symbols used for amino acids and peptides are in accordance with IUPAC-IUB Commission on Biochemical Nomenclature. Other abbreviations used are:
aa amino acid
Boc tert-butoxycarbonyl DCM dichloromethane
DICD N,N ' -diisopropylcarbodiimide
DIPE diisopropyl ether
DIPEA diisopropylethylamine
DMA dimethylacetamide
DMSO dimethylsulfoxide
EDT ethanedithiol
Fmoc 9-fluorenyl-methoxycarbonyl
HOBT N-hydroxybenzotriazole
MALDI Matrix assisted Laser Desoφtion Ionization
MS mass spectrometry
-Naeg- -[N-(2-acridin-9-ylamino)ethyl)glycinyl]-
(bivalent radical)
-Nahg- -[N-(6-aminohexyl)glycinyl]- (bivalent radical)
-Napg- -[N-(2-(4-(2-aminoethyl)phenoxy)ethyl glycinyl (bivalent radical)
-Naphg- -[N-(2-(amino)-2-(phenyl)-ethyl)-glycinyl]-
(bivalent radical)
-Narg- -[N-(3-guanidinopropyl)glycinyl]- (bivalent radical)
-Npeg- -[N-(2-(2-pyridyl)ethyl)-glycinyl]-
(bivalent radical)
-Nphe- -[N-(benzyl)glycinyl]- (bivalent radical)
-Nppg- -[N-(3-(2-oxopyπolidin-l-yl)propyl)glycinyl]-
(bivalent radical)
-NSG- -[N-substituted glycinyl]- (bivalent radical)
PE petroleum ether
Pme 2,2,5,7,8-pentamethylchroman-6-sulfonyl
TFA trifluoroacetic acid
TFE trifluoroethanol
TPTU 2-(pyridon- 1 -y 1)- 1 , 1 ,3 ,3-tetramethy luronium tetrafluroborate
Analytical HPLC is performed on a Shimadzu system (Kyoto, Japan) with product monitoring at 215 n by a Bischoff Lambda 1000 UV-detector from Metrohm (Wallisellen, Switzerland). A C18 reversed phase column (250 x 4.6 mm, Nucleosil C18, 5 μm; Macherey & Nagel, Duren, Germany) is used. The flow rate is 1 ml/min. A gradient system with two mobile phases is used: Mobile phase A = 0.1 % TFA in acetonitrile; mobile phase B = 0.1 % TFA in water. The linear gradient is from 10 % B/90 % A to 90 % B/10 % A in 30 min. The analytical data are presented as retention times tR.
Matrix-assisted laser desoφtion ionization (MALDI) mass spectrometry is carried out on a Linear Scientific LDI 1700 (Reno, NV, USA): the sample and 1,5-dihydroxybenzoic acid are co-crystallized, iπadiated with laser light (337 nm), and the masses of the ions produced are measured in a time-of-flight detection system.
Starting materials for the chemical synthesis of N-subsituted glycine derivatives and for solid phase synthesis are purchased from Fluka (Buchs, Switzerland), Aldrich (Steinheim, Germany), Sygena (Liestal, Switzerland), Novabiochem (Laufelfingen, Switzerland) and Bachem (Bubendorf, Switzerland). The Narg-building block (Fmoc-Narg(PMC)-OH = N-2,2,5,7,8-Pentamethylchroman-6-sulfonyl)-N'-3-(N-9-fluorenylmethoxycarbonyl- glycinyl)-propylguanidine) is synthesized as described in the literature (Heizmann et al., Peptide Research 7(6), 328-32 (1994).
The Examples 1 to 6 are obtained as trifluoroacetate salts.
Example 1 (SEQ ID NO: 1) H-Nahg-Narg-Narg-Nphe-Narg-(D-Lvs -(D-LvsV(D-ArgV(D-ProVNH
Resin 2 (see example lb)) is washed with ethylene chloride (3 x) and treated with 5 ml 95 % TFA/EDT (8:2) for 15 min. The treatment is repeated twice, while all filtrates are collected and pooled. The resin is washed with DCM (2 x) and TFE (2 x) and the filtrates are added to the pool, which is concentrated to a 5 ml volume in vacuo. 60 ml PE/DIPE (1:1) are added, and the filtrate is isolated and washed with PE/DIPE (1:1). The material is treated with 95% TFA/EDT (8:2) for 120 min, then precipitated again and washed with PE/DIPE (1:1). This procedure is repeated with a reduced reaction time of 60 min. The last precipitate is isolated, washed with PE/DIPE (1:1), dissolved in water and lyophilized. The crude product is purified (in 3 portions) by high performance liquid chromatography on a Nucleosil 7C18 (5 μm) Reversed Phase column (20 x 250 mm) (Macherey & Nagel, Duren, FRG): Eluent A = 0.1 % TFA/water, B = 0.1 % TFA acetonitrile; linear gredient 10 % B to 50 % B in 60 min; flow rate 18 ml/min; detection at 215 nm. The material eluted at the main peak is collected, lyophilized and obtained as a white powder. The product is characterised by mass spectrometry (electrospray MS, MALDI) and HPLC: MALDI-MS: molecule peak 1298 (corresponds to theory); HPLC: tR= 8.6 min.
The starting material is prepared as follows: l a) Fmoc-Narg('PmcVNarg(Pmc)-Nphe-Narg(Pmc)-D-Lvs(BocVD-Arg(PmcVD-Pro- amide of 4-(2,,4'-dimethoxyphenyl-aminomethyl)-phenoxymethyl-polvstyrene resin (Resin 1)
With 379 mg (0.085 μMol) Fmoc-D-Pro-ester of 4-(2',4'-dimethoxyphenyl-amino- methyl)-phenoxymethyl-polystyrene resin (prepared according to known procedures, see H. Rink, Tetrahedr. Lett. 28 (33), 3787-3790), the following procedure of alternating cleavage of the Fmoc group and (double) coupling of protected amino acid derivatives (Fmoc-D-Arg-Phe(Pmc)-OH, twice Fmoc-D-Lys(Boc)-OH, Fmoc-Narg(Pmc)-OH, Fmoc-Nphe-OH, twice Fmoc-Narg(Pmc)-OH in this sequence) is applied repetitively on an automated peptide synthesizer:
Washes (with approximately 10 ml) and reactions (while shaking in the reaction vessel) - 1 x 0.8 min isopropanol;
- 1 x preactivation for first coupling: 0.17 mMol (2 equiv.) Fmoc-aa are dissolved in 0.37 ml 0.5 M HOBT solution in DMA (2.2 equiv.) are added (in the case of Fmoc-Narg(Pmc), preactivation is made with TPTU instead of HOBT). During the activation time of 40 min further washes (as listed below) are caπied out:
- 2 x 0.4 min DMA;
- 12 x 0.5 min 20% piperidine in DMA (Fmoc cleavage);
- 2 x 0.4 min DMA;
- 1 x 0.8 min isopropanol;
- 5 x 0.3 min DMA;
- 1 x addition of the preactivation mixture described above.
- 1 x preactivation for the second part of the double coupling procedure: 0.17 mMol (2 equiv.) Fmoc-aa are dissolved in 0.37 ml 0.5 M HOBT solution in DMA (2.2 equiv.) and 0.09 ml 2 M DICD solution in DMA (2.2 equiv.) are added. During the activation time of 40 min, the first coupling is caπied out on the resin:
- 1 x 60 min coupling (first coupling); - 3 x 0.4 min DMA;
- 1 x addition of the preactivated mixture described above (second coupling); - 1 x 30 min coupling (second coupling);
- 2 x 0.3 min DMA;
- 1 x 4.5 min acetylation of unreacted amino groups with 10 ml acetic anhydride/pyridine/DMA (1:1:8);
- 5 x 0.3 min DMA;
- 1 x 0.8 min isopropanol; and
- 3 x 0.4 min DMA.
At the end of the chain assembly, the resin is washed 5 times with isopropanol and dried under vacuum.
l b) Nahg(Boc)-Narg(Pmc -Narg(PMC)-Nphe-Narg(Pmc -D-Lvs(BocVD-Lvs(Boc)- D-Arg(Pmc)-D-Pro-amide of 4-(2,,4'-dimethoxyphenyl-aminomethylVphenoxymethyl- polystyrene (Resin 2):
1.2 mMol (167 mg, 20 equiv.) of bromoacetic acid in 165 μl DMA are added to 1.32 mMol (206 μl, 22 equiv.) DICD in 340 μl DMA and preactivated for 10 min. The preactivated mixture is then added to resin 1 (example 1 a)). Then coupling is allowed during 30 min. The coupling is repeated with a second reagent mixture portion of the same composition as in the first coupling and allowed to proceed during 30 min. Then the following washing steps are made:
- 3 x 0.5 min DMA;
- 3 x 0.5 min DMSO;
- 4 mMol (66.7 equival.) of a 2.5 M solution of N-Boc-l,6-diaminohexane in DMSO are added to the resin and reacted for 150 min, followed by the following washing steps:
- 3 x 0.5 min DMSO;
- 3 x 0.5 min DMA; and 5 x 0.5 min isopropanol.
After drying in vacuo, Resin 2 is obtained.
The following examples 2 to 6 are prepared by methods analogous to those described in example 1:
Example 2: (SEQ ID NO:2): H-Nahg-Narg-Narg-Nphe-Narg-NH2
By deprotection/cleavage analogously as described for example 1, the title compound is obtained from the coπesponding starting material. MALDI-MS: molecule peak 789 (coπesponds to theory); HPLC: tR= 8.4 min.
Example 3: (SEQ ID NO:3 : H-Nahg-Narg-Narg-Nphe-Narg-(D-Lys)-NH?
By deprotection/cleavage analogously as described for example 1, the title compound is obtained from the coπesponding starting material. MALDI-MS: molecule peak 919.0 (theory: 919.07); HPLC: tR= 7.8 min.
Example 4: (SEQ ID NO:4 : H-Nahg-Narg-Narg-Nphe-Narg-(D-Lvs -(D-LvsVNH-,
By deprotection/cleavage analogously as described for example 1, the title compound is obtained from the coπesponding starting material. MALDI-MS: molecule peak 1047 (coπesponds to theory); HPLC: tR= 6.9 min.
Example 5: (SEQ ID NO:5 : H-Nahg-Narg-Narg-Nphe-Narg-(D-LysVω-LvsV(D-Arg)-NH7
By deprotection/cleavage analogously as described for example 1, the title compound is obtained from the coπesponding starting material. MALDI-MS: molecule peak 1201 (corresponds to theory); HPLC: tR= 7.9 min.
Example 6: (SEQ ID NO:6):
N-acetyl-Ser-Phe-Thr-Thr-Lvs-Ala-Leu-Glv-Ile-Ser-Tyr-Glv-Nahg-
Narg-Narg-Nphe-Narg-(D-Lvs)-(D-ArgV(D-Pro)-NH7
By deprotection/cleavage analogously as described for example 1, the title compound is obtained from the corresponding starting material. MALDI-MS: molecule peak 2566.4 (theory: 2567); HPLC: tR= 14.1 min.
Example 7: (SEQ ID NO:7) Nahg-Narg-Narg-Npeg-Narg-(D-Lys)-NH?:
Resin 1' is washed with ethylene chloride (3x) and treated with 5 ml 95% TFA/EDT (8:2) for 15 min. The treatment is repeated twice, while all filtrates are collected and pooled. The resin is washed with DCM (2x) and TFE (2x) and the filtrates are added to the pool, which is concentrated to a 5 ml volume in vacuo. 60 ml PE/DIPE (1:1) are added and the precipitate is isolated and washed with PE/DIPE (1:1). The material is treated with 95% TFA/EDT (8:2) for 120 min, then again precipitated ans washed with PE/DIPE (1:1). This procedure is repeated with a diminished reaction time of 60 min. The last precipitate is isolated, washed with PE/DIPE (1:1), dissolved in water and lyophilized. The crude product is purified with semi-preparative high performance liquid chromatography on a Nucleosil 7C18 (5 μm) reversed phase column (8 x 250 mm; Macherey & Nagel, Diiren, FRG): Eluent A = 0.1 % TFA/water, B = 0.1 % TFA/acetonitrile; linear gradient 1% B during 10 min, then 1% B to 50 % B in 30 min, flow 5 ml/min; detection at 215 nm. The material eluting with the main peak is collected, lyophilized and obtained as white powder. The product (title compound) is characterized by analytical HPLC with an analogous chromatography, using a Nucleosil 7C18 (5 μm) reversed phase column (4.6 x 250 mm, Macherey & Nagel, Duren, FRG), linear gradient 1% B to 90 % B in 30 min; flow 1 ml/min; detection at 215 nm; as well as by mass spectrometry (MALDI on an instrument from Linear Scientific, Reno, NV: model LDI 1700). HPLC-retention time: 11.3 min; MALDI-MS: molecule peak 934 (coπesponds to theory).
The starting material is prepared as follows:
7 a) Fmoc-Nahg(Boc)-Narg(PmcVNarg(PmcVNpeg-Narg(Pmc)-(D-Lvs)(Boc)-amide of 4-(2',4'-dimethoxyphenyl-aminomethyl)phenoxymethyl-polvstyrene (Resin 1 ')) With 50 mg (20 μmol) Fmoc-(D-Lys)(Boc)-amide of 4-(2',4'-dimethoxyphenyl-amino- methyl)phenoxymethyl-polystyrene resin (prepared according to knwon methods, see H. Rink, Tetrahedr. Lett. 28(33), 3787-90 (1987)), the following procedure A of alternating cleavage of the Fmoc group and (double) coupling of protected amino acid derivatives (see methods published by G. Heizmann and E. Felder, Peptide Res. 7, 328-32 (1994) is used for coupling Fmoc-(Narg(Pmc)). Procedure B is used for the incoφoration of Npeg and Nahg(Boc) with the primary amines 2-(pyridin-2-yl)ethylamine (Fluka, Buchs, Switzerland) and Boc-l,6-diaminohexane, respectively, following the principles of the submonomer approach (see Zuckermann, R.N., et al., J. Am. Chem. Soc. 114, 10646-7 (1992)):
Procedure A:
Washes (with approximately 10 ml) and reactions (while shaking the reaction vessel):
- lx 0.8 min isopropanol
- lx preactivation for first coupling: 0.04 mMol (2 equiv.) Fmoc-aa are dissolved in 0.09 ml 0.5 M TPTU solution in DMA (2.2 equiv.) and 0.022 ml 2M DICD solution in DMA (2.2 equiv.) are added. During the activation time of 40 min further washes (listed below) are carried out:
- 2x 0.4 min DMA
- 12x 0.5 min 20% piperidine in DMA (Fmoc cleavage)
- 2x 0.4 min DMA
- lx 0.8 min isopropanol
- 5x 0.3 min DMA
- lx addition of the preactivation mixture described above
- lx preactivation for the second part of th double coupling procedure: 0.04 mMol (2 equiv.) Fmoc-aa are dissolved in 0.09 ml 0.5 M TPTU solution in DMA (2.2 equiv.) and 0.022 ml 2M DICD solution in DMA (2.2 equiv.) are added. During the activation time of 40 min, the first coupling is carried out on the resin.
- lx 60 min coupling (first coupling)
- 3x 0.4 min DMA
- lx addition of the preactivated mixture described above (second coupling)
- lx 30 min coupling (second coupling)
- 2x 0.3 min DMA
- lx 4.5 min acetylation of unreacted amino groups with 10 ml acetic anhydride/pyridine/DMA (1:1:8)
- 5x 0.3 min DMA
- lx 0.8 min isopropanol
- 3x 0.4 min DMA
Procedure B:
0.4 mMol (55 mg, 20 equiv.) bromoacetic acid in 55 μl DMA are added to o.44 mMol (69 μl, 22 equiv.) DICD in 113 μl DMA and preactivated for 10 min. - the peactivated mixture is then added to the resin with the respective attached peptide derivatives;
- 30 min coupling
- repeat coupling with a second reagent mixture portion as above (30 min)
- 3x 0.5 min DMA
- 3x 0.5 min DMSO
- 1.32 mMol (66 equival.) of a 2.5 M solution of 2-(pyridin-2-yl)ethylamine (for Npeg) or Boc-l,6-diaminohexane (for Nahg) in DMSO is added to the resin and reacted for 150 min
- 3x 0.5 min DMSO
- 3x 0.5 min DMA
- 5x 0.5 min isopropanol.
After drying in vacuo, Resin 1' is obtained.
Example 8 (SEQ ID NO:8
Nahg-Narg-Narg-Nppg-Narg-(D-Lys)-NH?
By deprotection/cleavage analogously to that described for example 7, the title compound is obtained from the coπesponding starting materials (for introduction of Nppg,
3-(2-oxopyπolidin-l-yl)propylamin (Aldrich, Buchs, Schweiz) is used in Procedure B).
MALDI-MS: molecule peak 954 (coπesponds to theory);
HPLC: tR= 12.0 min.
Example 9 (SEQ ID NO:9
Nahg-Narg-Narg-Napg-Narg-(D-Lys)-NH?
By deprotection/cleavage analogously to that described for example 7, the title compound is obtained from the coπesponding starting materials (for introduction of Napg,
2-[4-(2-aminoethyl)phenoxy]-ethylamin is used in Procedure B).
MALDI-MS: molecule peak 992 (corresponds to theory);
HPLC: tR= 12.3 min.
The starting material is prepared as follows:
9 a) 2-[4-(2-Aminoethyl phenoxy]-ethylamin
49.1 g Boc-protected 2-(4-(2-aminoethyl)phenoxy)ethylbromide (is obtained by simple reaction of dibromoethane with Boc-protected 4-(2-aminoethyl)phenol which is obtained easily by Boc-protection of 4-(2-aminoethyl)phenol (Aldrich, Buchs, Switzerland) are reacted with 18.5 g sodium azide at 100 °C during 1 h. The reaction mixture is passed onto alkaline ice water. The suspension is extracted with ether/petrol ether (2 x), and the organic phases are washed with water. Evaporation leads to the title compound, melting point 56-65 °C.
Example 10 (SEQ ID NO: 10
Nahg-Narg-Narg-Naphg-Narg-(D-Lvs -NH2
By deprotection/cleavage analogously to that described for example 7, the title compound is obtained from the coπesponding starting materials (for introduction of Naphg,
2-(amino)-2-(phenyl)-ethylamin is used in Procedure B).
MALDI-MS: molecule peak 948 (corresponds to theory);
HPLC: tR= 12.1 min.
The starting material is prepared as follows:
10 a) 2-(Amino)-2-(phenyl)-ethylamin:
The title compound is obtained by catalytic hydrogenation of Boc-protected 2-(amino)-2-(phenyl)-acetonitrile (obtainable by first reacting benzaldehyde cyanhydrin (Aldrich, Buchs, Switzerland) in ethanol with 5 M ammonia at room temperature for 8 days, then evaporation, take up residue in ether, extract with diluted HCl, neutralisation of the extract with NaOH-solution which leads to precipitation, take up product again in ehter and repeat the extraction/precipitation which leads to crystalline 2-(amino)-2-(phenyl)acetonitrile, which is then protected by reaction with Boc-anhydride in tetrahydrofurane) at 105 °C/70 bar in tetrahydrofurane/triethylamine (30:1): melting point 89-92 °C.
Example 11 (SEQ ID NO: 11)
Nahg-Narg-Narg-Naeg-Narg-(D-Lys)-NH
By deprotection/cleavage analogously to that described for example 7, the title compound is obtained from the coπesponding starting materials (for introduction of Naeg,
2-(acridin-9-ylamino)-efhylamin is used in Procedure B).
MALDI-MS: molecule peak 1048 (corresponds to theory);
HPLC: tR= 14.5 min.
The starting material is preapared as follows
11 a) 2-(Acridin-9-ylamino)-ethylamin lg (4.68 mMol) of 9-chloro-acridine (Acros, New Jersey, USA) amd 25 ml (0.372 mMol) of 1,2-diaminoethane (Fluka, Buchs, Switzerland) are hated at 100 °C for 18 h. After cooling to room temperature, the reaction mixture is concentrated under vacuum and the product is isolated by means of flash-chromatography (Kieselgel 60, 40 -63 μm, Merck, Darmstadt, FRG); solvent system: methylene chloride/methanol/25% NH3 = 90:10:2) giving 0.57 g (2.41 mMol) of an oily product (the title compound). Retention time: 6.29 min (4.6 x 250 mm RP18-HPLC column, Macherey & Nagel; 1 ml min, A = 0.1 % TFA in H2O, B = 0.1% TFA in MeCN, gradient: 5% to 100% B in 25 min). FAB-MS: (M+H)+ = 238.
Example 12; Gelatine solution:
A sterile-filtered aqueous solution, with 20 % cyclodextrins as solubilisers, of one of the compounds of formula I mentioned in the preceding Examples (e.g. Example 1) as active ingredient, is so mixed under aseptic conditions, with heating, with a sterile gelatine solution containing phenol as preservative, that 1.0 ml of solution has the following composition:
active ingredient 3 mg gelatine 150.0 mg phenol 4.7 mg dist. water with 20 % cyclodextrins as solubilisers 1.0 ml
Example 13: Sterile dry substance for injection:
5 mg of one of the compounds of formula I mentioned in the preceding Examples (e.g. Example 1) as active ingredient are dissolved in 1 ml of an aqueous solution with 20 mg of mannitol and 20 % cyclodextrins as solubilisers. The solution is sterile-filtered and introduced under aseptic conditions into a 2 ml ampoule, deep-frozen and lyophilised. Before use, the lyophilisate is dissolved in 1 ml of distilled water or 1 ml of a physio¬ logical saline solution. The solution is administered intramuscularly or intravenously. This formulation can also be introduced into a twin-chambered injection ampoule.
Example 14: Nasal spray:
500 mg of finely ground (<5.0 μm) powder of one of the compounds of formula I men¬ tioned in the preceding Examples (e.g. Example 1) is suspended as active ingredient in a mixture of 3.5 ml of Myglyol 812® and 0.08 g of benzyl alcohol. The suspension is introduced into a container having a metering valve. 5.0 g of Freon 12® are introduced under pressure into the container through the valve. The "Freon" is dissolved in the Myglyol/benzyl alcohol mixture by shaking. The spray container contains approximately 100 single doses which can be administered individually.
Example 15: Film-coated tablets
The following ingredients are used for the preparation of 10 000 tablets each containing
100 mg of active ingredient:
active ingredient 1000 g corn starch 680 g colloidal silica 200 g magnesium stearate 20 g stearic acid 50 g sodium carboxymethyl starch 250 g water quantum satis
A mixture of one of the compounds of formula I mentioned in the preceding Examples (e.g. Example 1) as active ingredient, 50 g of corn starch and the colloidal silica is processed with a starch paste, made from 250 g of corn starch and 2.2 kg of demineralised water, to form a moist mass. This is forced through a sieve having a mesh size of 3 mm and dried at 45° for 30 min in a fluidised bed drier. The dry granules are pressed through a sieve having a mesh size of 1 mm, mixed with a pre-sieved mixture (1 mm sieve) of 330 g of corn starch, the magnesium stearate, the stearic acid and the sodium carboxymethyl starch, and compressed to form slightly biconvex tablets.
SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT:
(A) NAME: CIBA-GEIGY AG
(B) STREET: Klybeckstr. 141
(C) CITY: Basel
(E) COUNTRY: Switzerland
(F) POSTAL CODE (ZIP): 4002
(G) TELEPHONE: +41 61 69 11 11 (H) TELEFAX: + 41 61 696 79 76 (I) TELEX: 962 991
(ii) TITLE OF INVENTION: Antiviral Peptoid Compounds
(iii) NUMBER OF SEQUENCES: li
(iv) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: Patentin Release #1.0, Version #1.30 (EPO)
(2) INFORMATION FOR SEQ ID NO: 1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:!
(D) OTHER INFORMATION: /product=
"N-(6-aminohexyl)glycinyl"
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(ix) FEATURE:
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(ix) FEATURE:
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(ix) FEATURE:
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(ix) FEATURE:
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(B) LOCATIONS
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(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Pro-amide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:
Xaa Xaa Xaa Xaa Xaa Lys Lys Arg Xaa
1 5
(2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 5 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:! (D) OTHER INFORMATION: /product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
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(B) LOCATIONS
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(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
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(ix) FEATURE:
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(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:5
(D) OTHER INFORMATION:/product=
" (N-(3-guanidinopropyl)glycinyl)-amide"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:
Xaa Xaa Xaa Xaa Xaa 1 5
(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
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(ix) FEATURE:
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(D) OTHER INFORMATION:/product= "N-(benzyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys-amide (= D-Lys-NH2) "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:
Xaa Xaa Xaa Xaa Xaa Xaa
1 5
(2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 7 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
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(D) OTHER INFORMATION:/product=
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(ix) FEATURE:
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(ix) FEATURE:
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(ix) FEATURE:
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(ix) FEATURE:
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(ix) FEATURE:
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(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys-amide (= D-Lys-NH2) "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:
Xaa Xaa Xaa Xaa Xaa Lys Xaa
1 5
(2) INFORMATION FOR SEQ ID NO: 5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 8 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:1
(D) OTHER INFORMATION: /product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
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(ix) FEATURE:
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(B) LOCATIONS
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(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATIO :/product= "N-(benzyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:7
(D) OTHER INFORMATION:/product= "D-Lys"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Arg-amide (= D-Arg-NH2) "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:
Xaa Xaa Xaa Xaa Xaa Lys Lys Xaa 1 5
(2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 21 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION:1
(D) OTHER INFORMATION:/product= "N-acetyl-(L)-Ser"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:13
(D) OTHER INFORMATION:/product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:14
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:15
(D) OTHER INFORMATION: /product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION: 16
(D) OTHER INFORMATION:/product= "N-(benzyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION: 17
(D) OTHER INFORMATIO :/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:18
(D) OTHER INFORMATIO :/product= "D-Lys"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:19
(D) OTHER INFORMATION:/product= "D-Lys"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:20
(D) OTHER INFORMATION:/product= "D-Arg"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:21
(D) OTHER INFORMATIO :/product= "D-Pro-amide (= D-Pro-NH2 ) " ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 6:
Xaa Phe Thr Thr Lys Ala Leu Glv lie Ser Tyr Gly Xaa Xaa XaaXaa
1 5 * 10 15
Xaa Lys Lys Arg Xaa 20
(2) INFORMATION FOR SEQ ID NO: 7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:1
(D) OTHER INFORMATION:/product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATIO :/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATIO :/oroduct=
"N-(2-(2-pyridy 1 )ethyl)glycy1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys-amide (= D-Lys-NH2) " (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7:
Xaa Xaa Xaa Xaa Xaa Xaa 1 5
(2) INFORMATION FOR SEQ ID NO: 8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION: 1
(D) OTHER INFORMATION:/product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-(2-oxopyrrolidin-l-yl)propyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product= "D-Lys-amide (= D-Lys-NH2 ) " (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8:
Xaa Xaa Xaa Xaa Xaa Xaa
1 5
(2) INFORMATION FOR SEQ ID NO: 9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:1
(D) OTHER INFORMATION: /product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(2-(4-(2-aminoethyl)phenoxy)ethyl)-Gly"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product=
"N-^-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys-amide (= D-Lys-NH2) " (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9:
Xaa Xaa Xaa Xaa Xaa Xaa 1 5
(2) INFORMATION FOR SEQ ID NO: 10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:1
(D) OTHER INFORMATION:/product=
"N-(6-aminohexyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinophenyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATIO :/product=
"N-(2-(amino)-2-(phenyl)-ethyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product= "D-Lys-amide (= D-Lys-NH2 ) "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: Xaa Xaa Xaa Xaa Xaa Xaa 1 5
(2) INFORMATION FOR SEQ ID NO: 11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 6 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATION:1
(D) OTHER INFORMATION:/product=
"N-(6-aminohexy1)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(2-(acridin-9-ylamino)ethyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION:/product=
"N-(3-guanidinopropyl)glycinyl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site
(B) LOCATIONS
(D) OTHER INFORMATION: /product= "D-Lys-amide (= D-Lys-NH2) "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: Xaa Xaa Xaa Xaa Xaa Xaa
1 5

Claims

What is claimed is:
1. A peptoid compound of the formula I,
Rr(A)k-(X)m-(B)n-R2 (I)
wherein
k is 0 to 20, m is 3 to 10, n is 1 to 10,
Rj represents hydrogen, acyl or an amino-substituent other than acyl,
R2 represents an OH group, a C-terminal protecting group or a primary, secondary or tertiary amino group,
any A independently of the others being present represents a bivalent radical of an α-amino acid,
any X independently of the others being present represents a bivalent radical of the partial formula II,
Figure imgf000073_0001
wherein
each of R3, R4 and R5, independently of the others, represents hydrogen or a side chain of an α-amino acid other than glycine, or R3 and R4 together foπn an alkylene bridge and R5 is hydrogen, or R4 and R5 together form an alkylene bridge and R3 is hydrogen;
with the proviso, that a) at least one of these bivalent radicals has an R3 which is a side chain of an α-amino acid other than glycine and proline; b) that at least one R3 other than alkyl or benzyl is present; and c) that the bivalent radicals X together form a TAR-binding, transactivation-deficient oligopeptide analogue of the basic domain of the HIV Tat protein, and
any B independently of the others being present represents a bivalent radical of an α-amino acid,
or a salt thereof if a salt-forming group is present.
2. A compound of formula I according to claim 1 wherein
k is O to 15, m is 4 to 10, n is 1 to 10,
Rj is
hydrogen,
lower alkanoyl,
substituted lower alkanoyl wherein the substituents are independently selected from
- cycloalkyi with from 3 to 7 carbon atoms;
- aryl selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, each of which is unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, lower alkanoyl, sulfo, lower alkane- sulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxy¬ phosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
- carbamoyl;
- carbamoyl substituted at the nitrogen atom by one or two radicals selected from lower alkyl, carboxy-lower alkyl or lower alkoxy- carbonyl-lower alkyl;
- heterocyclyl selected from thienyl, furyl, pyπolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, chromanyl, cyclohexa[b]pyπolyl, cyclohexa- [b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyπolidinyl, pyπolinyl, imidazolidyl, piperidyl, piperazinyl, morpholinyl, thiomo holinyl, S,S-dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by one or more substituents selected from lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxy¬ carbonyl, halogen, lower alkanoyl, nitro, oxo and cyano;
- hydroxy;
- lower alkoxy;
- lower alkanoyloxy;
- up to 3 halogen atoms and
- carboxy,
lower alkoxycarbonyl,
2-halo-lower alkoxycarbonyl,
aryl-lower alkoxycarbonyl wherein aryl is phenyl, 1- or 2-naphthyl, fluorenyl or phenyl mono- or poly-substituted by lower alkyl, phenyl, hydroxy, lower alkoxy, halogen or nitro,
heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is thienyl, furyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, iso- quinolyl, 3,1-benzofuranyl, cyclohexa[b]pyπolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyπolidinyl, pyπolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S- dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxycarbonyl, halogen, lower alkanoyl, nitro or cyano,
lower alkenyloxycarbonyl wherein the lower alkenyl radical is bonded to the bonding oxygen atom via a saturated carbon atom,
lower alkoxy-lower alkoxycarbonyl,
(lower alkoxy-lower alkoxy )-lower alkoxycarbonyl,
aryl-lower alkyl with up to three aryl groups, wherein each aryl is selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl which are unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di- lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, lower alkanoyl, sulfo, lower alkanesulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxyphosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
arylthio or aryl-lower alkylthio, wherein aryl is phenyl that is unsubstituted or substituted by lower alkyl, lower alkoxy, halogen or nitro,
1 -lower alkanoyl-lower alk-l-en-2-yl, lower alkoxycarbonyl-lower alk-l-en-2-yl,
tri-lower alkylsilyl,
arylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups,
(mono-, di- or tri-aryl)-lower alkylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups, or
2,2,5,7,8-pentamethylchroman-6-sulfonyl,
hydroxy,
a lower alkoxy group that is branched in the 1-position,
arylmethoxy having one or two aryl radicals, wherein aryl is phenyl that is unsubstituted or mono-, di- or tri-substituted by lower alkyl, lower alkoxy, hydroxy, halogen or nitro,
1 -lower alkoxy-lower alkoxy, 1 -lower alkylthio-lower alkoxy,
benzoylmethoxy wherein benzoyl is unsubstituted or substituted by halogen,
2-halo-lower alkoxy,
2-(tri-substituted silyl)-lower alkoxy wherein the substituents are each independently of the others selected from lower alkyl, phenyl-lower alkyl, C3-C7-cycloalkyl or phenyl,
tri-lower alkylsilyloxy, amino, or
a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyπolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, moφholinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyπolidinyl an 4-moφholinyl;
any A being present and any B being present are independently selected from the group comprising a bivalent radical, bonded via its α-amino and its α-carbonyl group, of an amino acid selected from glycine alanine, valine, norvaline, leucine, isoleucine, norleucine, α-amino-n-decanoic acid, serine, homoserine, threonine, methionine, cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine, tyrosine, 4-amino- phenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenyl- alanine, β-phenylserine,phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan, indoline-2-carboxylic acid, 1,2,3,4-tetrahydroiso- quinoline-3-carboxylic acid, aspartic acid, asparagine, aminomalonic acid, amino¬ malonic acid monoamide, glutamic acid, glutamine, histidine, arginine, lysine, N6-benzyl-Ne-methyl-lysine, N6,Ne-dibenzyl-lysine, δ-hydroxylysine, ornithine, α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyπolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino-γ-(4-moφholinyl), α-amino-γ-(3-indolyl)-butyric acid, α,γ-diaminobutyric acid and α,β-diamino- propionic acid; it being possible for each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form,
and each X being present is a bivalent radical of the partial formula II,
Figure imgf000078_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
- amino-lower alkyl,
- N-lower alkylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, N,N-di(phenyl-lower alkyl)-amino- or guanidino-lower alkyl,
- pyridyl-lower alkyl,
- pyπolidinyl-lower alkyl,
- moφholinyl-lower alkyl,
or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus, may in addition be a side chain selected from
- aryl-lower alkyl wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
- carboxy-lower alkyl, - carbamoyl-lower alkyl
- furyl-lower alkyl, and
- indolyl-lower alkyl,
or is selected from (acτidin-9-yl)amino-lower alkyl, (3,6-bis(dimethyl- amino)acridin-9-yl)amino-lower alkyl, 2-(6-chloro-9-methoxyacridin-9-yl)- amino-lower alkyl, (amino)-(phenyl)-lower alkyl, (amino-loweralkyl- phenoxy)-lower alkyl, and oxopyπolidinyl-lower alkyl;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than lower alkyl or benzyl;
and
R4 and R5 each represent hydrogen; or a salt thereof.
3. A compound of formula I according to claim 1 wherein
k is 0 to 15, m is 4 to 10, n is 1 to 10,
R, is
hydrogen,
lower alkanoyl,
substituted lower alkanoyl wherein the substituents are independently selected from
- cycloalkyi with from 3 to 7 carbon atoms;
- aryl selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl, each of which is unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di-lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, lower alkanoyl, sulfo, lower alkane- sulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxy¬ phosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
- carbamoyl;
- carbamoyl substituted at the nitrogen atom by one or two radicals selected from lower alkyl, carboxy-lower alkyl or lower alkoxy- carbonyl-lower alkyl;
- heterocyclyl selected from thienyl, furyl, pyπolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, isoquinolyl, 3,1-benzofuranyl, chromanyl, cyclohexa[b]pyrrolyl, cyclohexa- [b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyπolidinyl, pyπolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S-dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1, 2,3,4- tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by one or more substituents selected from lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxy¬ carbonyl, halogen, lower alkanoyl, nitro, oxo and cyano;
- hydroxy;
- lower alkoxy;
- lower alkanoyloxy;
- up to 3 halogen atoms and
- carboxy,
lower alkoxycarbonyl,
2-halo-lower alkoxycarbonyl,
aryl-lower alkoxycarbonyl wherein aryl is phenyl, 1- or 2-naphthyl, fluorenyl or phenyl mono- or poly-substituted by lower alkyl, phenyl, hydroxy, lower alkoxy, halogen or nitro,
heterocyclyl-lower alkoxycarbonyl wherein heterocyclyl is thienyl, furyl, pyπolyl, imidazolyl, pyrazolyl, oxazolyl, thiazolyl, tetrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolyl, benzimidazolyl, quinolyl, iso- quinolyl, 3,1-benzofuranyl, cyclohexa[b]pyπolyl, cyclohexa[b]pyridyl, cyclohexa[b]pyrazinyl, cyclohexa[b]pyrimidinyl, pyπolidinyl, pyπolinyl, imidazolidyl, piperidyl, piperazinyl, moφholinyl, thiomoφholinyl, S,S- dioxo-thiomoφholinyl, indolinyl, isoindolinyl, 4,5,6,7-tetrahydroindolyl, 1,2,3,4-tetrahydroquinolyl or 1,2,3,4-tetrahydroisoquinolyl, each of which is unsubstituted or substituted by lower alkyl, phenyl, 1- or 2-naphthyl, phenyl-lower alkyl, hydroxy-lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkoxycarbonyl, phenyl- or naphthyl-lower alkoxycarbonyl, halogen, lower alkanoyl, nitro or cyano,
lower alkenyloxycarbonyl wherein the lower alkenyl radical is bonded to the bonding oxygen atom via a saturated carbon atom,
lower alkoxy-lower alkoxycarbonyl,
(lower alkoxy-lower alkoxy)-lower alkoxycarbonyl,
aryl-lower alkyl with up to three aryl groups, wherein each aryl is selected from phenyl, indenyl, indanyl, naphthyl, anthryl, phenanthryl, acenaphthyl or fluorenyl which are unsubstituted or mono- to tri-substituted by lower alkyl, halo-lower alkyl, phenyl, 1- or 2-naphthyl, hydroxy, lower alkoxy, carbamoyl-lower alkoxy, N-lower alkylcarbamoyl-lower alkoxy, N,N-di- lower alkylcarbamoyl-lower alkoxy, amino, mono- or di-lower alkylamino, lower alkanoylamino, halogen, carboxy, lower alkoxycarbonyl, phenyl-, naphthyl- or fluorenyl-lower alkoxycarbonyl, lower alkanoyl, sulfo, lower alkanesulfonyl, phosphono, hydroxy-lower alkoxyphosphoryl, di-lower alkoxyphosphoryl, carbamoyl, mono- or di-lower alkylcarbamoyl, sulfamoyl, mono- or di-lower alkylaminosulfonyl, nitro or cyano;
arylthio or aryl-lower alkylthio, wherein aryl is phenyl that is unsubstituted or substituted by lower alkyl, lower alkoxy, halogen or nitro,
1 -lower alkanoyl-lower alk-l-en-2-yl,
lower alkoxycarbonyl-lower alk-l-en-2-yl,
tri-lower alkylsilyl,
arylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups,
(mono-, di- or tri-aryl)-lower alkylsulfonyl, wherein aryl is phenyl that is unsubstituted or substituted by one to five lower alkyl or lower alkoxy groups, or 2,2,5,7,8-pentamethylchroman-6-sulfonyl,
R-. is
hydroxy,
a lower alkoxy group that is branched in the 1 -position,
arylmethoxy having one or two aryl radicals, wherein aryl is phenyl that is unsubstituted or mono-, di- or tri-substituted by lower alkyl, lower alkoxy, hydroxy, halogen or nitro,
1-lower alkoxy-lower alkoxy, 1-lower alkylthio-lower alkoxy,
benzoylmethoxy wherein benzoyl is unsubstituted or substituted by halogen,
2-halo-lower alkoxy,
2-(tri-substituted silyl)-lower alkoxy wherein the substituents are each independently of the others selected from lower alkyl, phenyl-lower alkyl, C3-C7-cycloalkyl or phenyl,
tri-lower alkylsilyloxy,
amino, or
a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyπolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, moφholinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1 -pyπolidinyl an 4-moφholinyl;
any A being present and any B being present are independently selected from the group comprising a bivalent radical, bonded via its α-amino and its α-carbonyl group, of an amino acid selected from glycine alanine, valine, norvaline, leucine, isoleucine, norleucine, α-amino-n-decanoic acid, serine, homoserine, threonine, methionine, cysteine, proline, trans-3- and trans-4-hydroxyproline, phenylalanine, tyrosine, 4-amino- phenylalanine, 4-nitrophenylalanine, 4-chlorophenylalanine, 4-carboxyphenyl- alanine, β-phenylserine,phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, tryptophan, indoline-2-carboxylic acid, 1,2,3,4-tetrahydroiso- quinoline-3-carboxylic acid, aspartic acid, asparagine, aminomalonic acid, amino¬ malonic acid monoamide, glutamic acid, glutamine, histidine, arginine, lysine, Ne-benzyl-Ne-methyl-lysine, Ne,Ne-dibenzyl-lysine, δ-hydroxylysine, ornithine, α-amino-β-(2-furyl)-propanoic acid, α-amino-γ-(l-pyπolidinyl)-butyric acid, α-amino-γ-(2-pyridyl)-butyric acid, α-amino-γ-(4-moφholinyl), α-amino-γ-(3-indolyl)-butyric acid, α,γ-diaminobutyric acid and α,β-diamino- propionic acid; it being possible for each of the mentioned amino acids (with the exception of glycine) to be in the D-, L- or (D,L)-form,
and each X being present is a bivalent radical of the partial formula II,
Figure imgf000084_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
- amino-lower alkyl,
- N-lower alkylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, N,N-di(phenyl-lower alkyl)-amino- or guanidino-lower alkyl,
- pyridyl-lower alkyl,
- pyπolidinyl-lower alkyl,
- moφholinyl-lower alkyl, or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus, may in addition be a side chain selected from
- aryl-lower alkyl wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
- carboxy-lower alkyl, - carbamoyl-lower alkyl
- furyl-lower alkyl, and
- indolyl-lower alkyl;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than lower alkyl or benzyl;
and
R4 and Rg each represent hydrogen;
or a salt thereof.
4. A compound of formula I according to claim 1, wherein
k is 0 to 12, m is 4 to 6, n is 1 to 4;
Rj is
hydrogen or
lower alkanoyl, such as acetyl,
R, is amino, or further a mono- or disubstituted amino group the substituents of which are selected independently from the group comprising lower alkyl, phenyl-lower alkyl, pyπolidinyl-lower alkyl, pyridyl-lower alkyl, furyl-lower alkyl, moφholinyl-lower alkyl and indolyl-lower alkyl, or a disubstituted amino group selected from 1-pyπolidinyl an 4-moφholinyl;
any A being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or L-form (where an asymmetric α-carbon atom is present);
any B being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
and any X being present is a bivalent radical of the partial formula II,
Figure imgf000086_0001
wherein
R3 represents a side chain selected from the group comprising lower alkyl or substituted lower alkyl selected from
- amino-lower alkyl,
- N-lower alkylamino-, N,N-di-lower alkylamino-, N-(phenyl-lower alkyl)-N-(lower alkyl)-amino-, N,N-di(phenyl-lower alkyl)-amino- or guanidino-lower alkyl,
- pyridyl-lower alkyl,
- pyrrolidinyl-lower alkyl, and
- moφholinyl-lower alkyl,
or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus, may in addition be a side chain selected from
- aryl-lower alkyl wherein aryl is phenyl that is unsubstituted or mono- or poly-substituted by lower alkyl, lower alkoxy, lower alkanoyloxy, amino, lower alkylamino, di-lower alkylamino, lower alkanoylamino, lower alkoxycarbonylamino, arylmethoxycarbonylamino wherein aryl has from 6 to 14 carbon atoms, halogen, carboxy or nitro;
- carboxy-lower alkyl,
- carbamoyl-lower alkyl,
- furyl-lower alkyl and
- indolyl-lower alkyl;
with the proviso that at least one bivalent radical of the formula II is present wherein R3 is other than lower alkyl or benzyl;
and R4 and R5 each represent hydrogen;
or a salt thereof.
5. A compound of formula I according to claim 1 wherein
k is 0 or 12, m is 4 to 6, n is 1 to 4;
Rj is
hydrogen or
lower alkanoyl,
R, is amino,
any A being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from glycine, alanine, leucine, isoleucine, phenylalanine, tyrosine, serine, threonine and lysine, which are present in the D- or preferably in the L-form;
any B being present is a bivalent radical of an α-amino acid bound at its N-terminus via its α-amino group and at its C-terminus via its α-carbonyl group selected from lysine, arginine and proline, each of which is present preferably in the D-form;
and any X being present is a bivalent radical of the partial formula II,
Figure imgf000088_0001
wherein
R3 represents a side chain selected from the group comprising
- amino-lower alkyl and
- guanidino-lower alkyl, or, in the case of the residue X which is in position 4 of the bivalent residue -(X)m- when counted from the N-terminus, may in addition be a phenyl-lower alkyl side chain,
and R4 and R5 each represent hydrogen;
or a salt thereof.
6. A peptoid compound of formula I according to claim 1 with the designation H-Nahg-Narg-Narg-Nphe-Narg-(D-Lys)-(D-Lys)-(D-Aτg)-(D-Pro)-NH2 (SEQ. ID NO:l) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical -[N-(benzyl)glycinyl]-, or a salt thereof.
7. A peptoid compound of formula I according to claim 1 with the designation H-Nahg-Narg-Narg-Nphe-Narg-(D-Lys)-NH2 (SEQ. ID NO:3) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical -[N-(benzyl)glycinyl]-, or a salt thereof.
8. A peptoid compound of formula I according to claim 1 with the designation H-Nahg-Narg-Narg-Nphe-Narg-(D-Lys)-(D-Lys)-NH2 (SEQ. ID NO:4) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical -[N-(benzyl)glycinyl]-, or a salt thereof.
9. A peptoid compound of formula I according to claim 1 with the designation H-Nahg-Narg-Narg-Nphe-Narg-(D-Lys)-(D-Lys)-(D-Arg)-NH2 (SEQ. ID NO:5) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical -[N-(benzyl)glycinyl]-, or a salt thereof.
10. A peptoid compound of formula I according to claim 1 with the designation N-acetyl-Ser-Phe-Thr-Thr-Lys-Ala-Leu-Gly-Ile-Ser-Tyr-Gly-Nahg- Narg-Narg-Nphe-Narg-(D-Lys)-(D-Arg)-(D-Pro)-NH2 (SEQ. ID NO:6) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical -[N-(benzyl)glycinyl]-, or a salt thereof.
11. A peptoid compound of formula I according to claim 1 with the designation Nahg-Narg-Narg-Npeg-Narg-(D-Lys)-NH2 (SEQ ID NO:7) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Npeg- is the bivalent radical -[N-(2-(2-pyridyl)ethyl)-glycinyl]-, or a salt thereof.
12. A peptoid compound of formula I according to claim 1 with the designation Nahg-Narg-Narg-Nppg-Narg-(D-Lys)-NH2 (SEQ D NO:8) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nppg- is the bivalent radical -[N-(3-(2-oxopyπolidin-l-yl)propyl)glycinyl]-, or a salt thereof.
13. A peptoid compound of formula I according to claim 1 with the designation Nahg-Narg-Narg-Napg-Narg-(D-Lys)-NH2 (SEQ D NO:9) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Napg- is the bivalent radical -[N-(2-(4-(2-aminoethyl)phenoxy)ethyl)-glycinyl, or a salt thereof.
14. A peptoid compound of formula I according to claim 1 with the designation Nahg-Narg-Narg-Naphg-Narg-(D-Lys)-NH2 (SEQ D NO:10) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinylj- and -Naphg- is the bivalent radical -[N-(2-(amino)-2-(phenyl)-ethyl)-glycinyl]-, or a salt thereof.
15. A peptoid compound of formula I according to claim 1 with the designation Nahg-Narg-Narg-Naeg-Narg-(D-Lys)-NH2 (SEQ D NO: 11) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Naeg- is the bivalent radical -[N-(2-acridin-9-ylamino)ethyl)glycinyl]-, or a salt thereof.
16. A pharmaceutical preparation comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15 or 20 and a caπier material.
17. A compound of formula I, or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 15 or 20 for use in a method for the therapeutic or prophylactic treatment of the warm-blooded animal or human body.
18. The use of a compound of formula I, or a salt thereof, according to any one of claims 1 to 15 or 20 for the preparation of a pharmaceutical composition for the treatment of a retroviral infection.
19. A process for the manufacture of a compound of formula I according to claim 1, said process comprising
a) reacting a fragment of a compound of formula I which has a free carboxy group, or a reactive derivative thereof, with a complementary fragment that has an amino group with at least one free hydrogen atom, or with a reactive derivative thereof, with formation of an amide bond; in the mentioned fragments free functional groups with the exception of those that participate in the reaction if required being present in protected form; and removing any protecting groups present; or
b) for the synthesis of compounds of formula I wherein k = 0, Rj is hydrogen and the N-terminal X is a bivalent radical of the partial formula II,
Figure imgf000091_0001
wherein R3 is a residue from the group comprising lower alkyl or substituted lower alkyl, while R2, R4, R5, A, B, X, m and n have the meanings given for compounds of the formula I,
alkylating an amino compound of the formula IV,
H2N-R3 (IV),
wherein R3 has the meanings given above, with a compound of the formula V
Z-C(R4R5)-C(=O)-(X)m.]-(B)n-R2' (V)
wherein Z is a nucleofugal leaving group, R2' has the same meaning as R2 in compounds of formula I or is a resin for solid phase synthesis and R , R5, X, B, m and n have the meanings given for compounds of formula I, under nucleophilic substitution, in the mentioned starting materials free functional groups with the exception of those that participate in the reaction if required being present in protected form; and removing any protecting groups and cleaving from any resin for solid phase synthesis being present;
and, if desired, transforming a salt of an obtainable compound of formula I into the free compound or a different salt or an obtainable free compound of formula I into a salt, and/or separating obtainable mixtures of isomers of compounds of formula I into the individual isomers.
20. A peptoid compound with the designation
H-Nahg-Narg-Nphe-Narg-NH2 (SEQ. ID NO:2) wherein -Nahg- is the bivalent radical -[N-(6-aminohexyl)glycinyl]-, -Narg- is the bivalent radical -[N-(3-guanidinopropyl)glycinyl]- and -Nphe- is the bivalent radical
-[N-(benzyl)glycinyl]-, or a salt thereof.
PCT/EP1996/002424 1995-06-07 1996-06-04 Antiviral peptoid compounds Ceased WO1996040759A1 (en)

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WO1999002488A1 (en) * 1997-07-07 1999-01-21 University Of Medicine And Dentistry Of New Jersey Inhibition of hiv-1 replication using oligocarbamate derivatives
WO1999025327A3 (en) * 1997-11-14 1999-09-23 Warner Lambert Co Small molecule intervention in hiv-1 replication
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US10087221B2 (en) 2013-03-21 2018-10-02 Sanofi-Aventis Deutschland Gmbh Synthesis of hydantoin containing peptide products
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