JP2002530430A - Pharmaceutical composition for inhibiting hepatitis C virus NS3 protease - Google Patents

Abstract

  (57) [Summary] Disclosed are peptide inhibitors of the hepatitis C virus N53 protease based on the P and P 'regions of the natural substrate. The P 'portion of the inhibitor is optimized to achieve maximum binding energy by interaction with the S' region of the enzyme. By selecting amino acids such that the inhibitor cannot be substantially cleaved by NS3 protease, an NS3 protease inhibitor having a titer in the low nanomolar range up to the nanomolar range can be obtained.

Description

DETAILED DESCRIPTION OF THE INVENTION

TECHNICAL FIELD [0001] The present invention relates to compounds that can act as inhibitors of the hepatitis C virus (HCV) NS3 protease, and the use of such compounds and their preparation.

BACKGROUND ART Hepatitis C virus (HCV) is a parenteral, sporadic non-A, non-B hepatitis (N
ANB-H). It is thought that approximately 1% of the population worldwide is affected. Viral infections can cause chronic hepatitis and cirrhosis, thereby causing hepatocellular carcinoma. Currently, treatment with recombinant interferon alpha alone or in combination with ribavirin has been partially successful in a small number of cases,
No vaccine or treatment has been established. Thus, there is an urgent need for new and broadly effective therapeutic agents.

[0003] Some virally encoded enzymes are metalloproteases (NS2-3).
, Serine proteases (NS3), helicases (NS3), and RNA-dependent RNA polymerases (NS5B) are putative targets for therapeutic intervention. The NS3 protease is located in the N-terminal domain of the NS3 protein, with intramolecular cleavage at the NS3 / 4A site and NS4A / 4B, NS4B / 5A, and NS5A / 5B.
It is considered a major drug target because it is responsible for downstream intramolecular processing at the junction.

[0004] Previous studies have identified a group of peptides, particularly hexapeptides, that exhibit some NS3 protease inhibitory activity. It is an object of the present invention to provide further compounds which exhibit similar and possibly improved activity.

[0005] Schechter & Berger (1967, Biochem. Bio)
phys. Res. Commun. 27, 157-162), the cleavage site in the NS3 protease substrate is P6-P5-P4-P3-P2-P
1 ... P1′-P2′-P3′-P4 ′-(P represents an amino acid,
There is an easily cleavable bond between P1 and P1 ′). The corresponding enzyme binding sites are S6-S5-S4-S3-S2-S1 ... S1'-S2
It is called '-S3'-S4'-.

[0006] Applicants have previously disclosed so-called product inhibitors that are optimized for low nanomolar titers based on the P region of the natural cleavage site (1998, Bioc).
chemistry, 37, 8899-8905 and 1998, Biochem.
illustration, 37, 8906-8914). These inhibitors draws much of its binding energy from the C-terminal carboxylic acid, the interaction of the left NS3, similar to the interaction used by the natural substrates, including, in S ₁ pocket Significant electrostatic interactions of the bond and the P6-P5 acidic bond are involved.

[0007] Unlike the P region, the P 'region of the substrate, while important for catalysis, does not significantly affect ground state binding to the enzyme, expressed as a Km value. In other words, the binding energy released by the interaction of the substrate with the enzyme to form an initial non-covalent complex is essentially due to the interaction of residues in the P region,
The residues in the 'region contribute to the binding energy to a lesser extent. Therefore, '(over to P _10' residues P ₁₎ peptide based on the region 'P of the natural substrate do not inhibit NS3 in any significant extent. Despite this, inspection of the crystal structure of NS3 with or without 4A (and more recently the NMR structure of NS3) shows that
The presence of a binding pocket in the S 'region that could be used for binding inhibitors directed to the active site was demonstrated. Thus, S 'binding ligands exhibit a range of interactions with enzymes different from those used by the substrate, and represent a new class of NS3 inhibitors.

[0008] Landro et al., 1997, Biochemistry, 36, 9340-9.
348 is defined by replacing P ₁ 'serine with a bulky cyclic aromatic compound (tetrahydroisoquinoline-3-carboxylic acid) or a smaller cyclic alkyl compound (proline or pipecolic acid) based on the NS5A / 5B cleavage site. Of the uncleaved decapeptides were synthesized. They then investigated the interaction of this decapeptide with the NS3 substrate binding site, either in the presence or absence of the NS4A cofactor. A truncation effect at either the P or P 'site of the molecule (effect
By investigating ct of truncation, they concluded that most of the binding energy of the decapeptide was due to interaction with the NS3-NS4A complex on the P-side of the molecule. Pruning on the P 'side has a relatively large effect in the presence of the NS4A cofactor, but is less effective in the absence of NS4A. They are,
If the Tyr residue of the P4 'substrate present in the molecule is closely proximal or NS4A
It was concluded that this residue significantly contributed to binding in the presence of NS4A.

The present inventors have developed inhibitors that are more potent than those described by Landro et al. Because they bind better on their P ′ side. In other words,
This inhibitor takes advantage of binding to the S 'region in addition to binding to the NS3 S region. By changing the P 'amino acid residues, we can derive from binding of the S' region because inhibitors with optimized or non-optimized P 'regions are more than 1000-fold more potent. It has been shown that the binding energy is substantial. 'Because there is no activity in any of the peptides corresponding to the region, S' isolated P optimization of binding fragments, were explored in the range of P ₆ uncut decapeptide over P _{4 '.}

The present inventors have proposed that by replacing the P4′Tyr residue of Landro with leucine,
It has been found that the effectiveness of the decapeptide as an NS3 protease inhibitor can be improved. In a decapeptide substrate cleavable by PNS3,
Leucine at position 4 'has been previously shown to be better than tyrosine (Urbani et al., 1997, J. Biol. Chem., 272, 9).
204-9209), which is the first indication that this substitution has been applied to a decapeptide inhibitor that is not cleaved under the influence of the enzyme. P4 'residue and P2'-P
By optimizing the 3 'fragments and using them with the optimized P region, we have reached oligopeptides that display titers in the low nanomolar to subnanomolar range.

According to a first aspect of the present invention, there is provided a compound having formula (I). Pep-A'-B'-C'-D '(I) (N-terminal on left and C-terminal on right) wherein "Pep" is a peptide or a peptide capable of binding to HCV NS3 protease. A ′ is, for example, an optionally substituted proline residue having 1 to 3 substituents, wherein B ′ is An amino acid or amino acid analog having a non-polar side chain, preferably this side chain is an alkyl, aryl or aralkyl group containing 3 to 10, especially 4 to 8, carbon atoms; An amino acid or amino acid analog having a polar side chain, examples of polar side chains may include 2-10, preferably 2-6, carbon atoms; D ′ is leucine, less preferred but non-polar Aliphatic side Another amino acid (such as valine, isoleucine, norleucine, or methionine) or a short peptide or peptide analog having one of these amino acids (particularly N-terminal leucine); Can contain, for example, 2 to 6, preferably 2 to 4 amino acids or amino acid analogs.

As used herein, the term “amino acid analog” refers to, for example, α-
And organic compounds that are not necessarily naturally occurring.

The Prep-A bond of the compound of formula (I) is substantially non-cleavable by the HCV NS3 protease. Preferably, cleavage is undetectable using, for example, the assay described under the heading "Substrate Assay" below.

[0014] Pharmaceutically acceptable salts and derivatives (such as esters) of the compounds of formula (I) are within the scope of the invention.

Preferably, the compound of formula (I) is acylated, especially acetylated, at the N-terminus, but other derivatives of the N-terminus (eg, N-terminal sulfoxide, sulfonamide,
Urethane or urea derivatives) are also possible.

Preferably, the compound of formula (I) is amidated at the C-terminus. But C
The terminus may be a non-derivatized carboxylic acid group. Alternatively, other C-terminal groups may be present.

Assuming that there is no proline residue substitution at A ′, a preferred C-terminal portion of the compound of formula (I) is Pro-B′-C′-Leu (possibly with a short C-terminus at Leu ).

In the compound of the first aspect of the present invention, preferred examples of the amino acid or analog contained in B ′ include β-cyclohexylalanine, phenylglycine, homophenylalanine, and norleucine, and more preferably less. However, other possibilities are leucine, methionine, norvaline, and β-cyclopropylalanine. Of all these, cyclohexylalanine and phenylglycine are most preferred.

As examples of amino acids or analogs, C ′ includes aspartic acid, glutamic acid, γ-carboxyglutamic acid, glutamine, asparagine, and hydroxyproline. N-β-Aloc-diaminobutyric acid, thiazolylalanine,
Methionine sulfoxide, pyridylalanine, and serine are slightly less preferred. Of all these, aspartic acid is most preferred.

The following combinations of amino acid residues at B ′ and C ′ are preferred, and a combination of cyclohexylalanine and aspartic acid is particularly preferred.

[0021]

[Table 1] Footnote: Cha = β-cyclohexylalanine. Nle = norleucine. Phg = phenylglycine. Hof = homophenylalanine. Hyp = hydroxyproline.

In the case of leucine (or other amino acids) where residue D 'has a small peptide as a C-terminal extension, the peptide can be selected relative to the corresponding P' portion of the natural substrate.

[0023] Residues A ', B', C ', and D' can have D- or L-stereochemistry, but in general, each L-stereochemistry is preferred.

With respect to the Pep moiety of the compound of formula (I), this means, for example, that if Pep carries only a carboxylic acid group at the C-terminus, the non-C-terminal residues A′-B′-C′-D ′ Peptides or peptide analogs that can bind to HCV NS3 protease even in the presence are preferred. For example, the fragment Pep-OH preferably has an IC ₅₀ of less than 100 μM (eg, less than 20 μM, especially less than 10 μM, optimally less than 1 μM) when tested in the following inhibition assays. Preferably, P
ep is a hexa, penta, or tetrapeptide having the formula II FEDDCBA (II) below. Wherein A is an amino acid or amino acid analog having a relatively small (C ₁ -C ₆ ) aliphatic side chain. Possible choices for this group include cysteine, aminobutyric acid (Abu) (including di and trifluoro Abu), norvaline, allylglycine, and alanine, each of which can be N-methylated. Of these, cysteine and fluorinated aminobutyric acid are preferred choices for A.

B is an amino acid or analog having a non-polar or acidic side chain. Some amino acids having polar but uncharged side chains may also be suitable. Examples of suitable amino acids include glutamic and aspartic acid, glycine and methylglycine, 2-aminobutyric acid, alanine, isoleucine, valine, leucine, cysteine, naphthylalanine, and β-cyclohexylalanine. Of these, cyclohexylalanine is particularly preferred.

C is an amino acid or amino acid analog having a non-polar or acidic side chain. For example, as an example of such an amino acid, the amino acid shown in B above is also applied to C. In this case, isoleucine and glutamic acid are particularly preferred.

D is an amino acid or amino acid analog having a hydrophobic side chain (such as methionine, isoleucine, leucine, norleucine, valine, methylvaline, phenylglycine, or diphenylalanine). Of these, leucine and especially diphenylalanine are preferred. Some polar amino acids containing a hydrophobic moiety, such as tyrosine, thienylalanine, and chlorophenylalanine, may be suitable.

E may not be present with F, but if it is, it is generally an amino acid or amino acid analog having an acidic side chain. Preferred examples are glutamic acid and aspartic acid, the former being preferred. Alternatively, E can be an amino acid or analog having a nonpolar or polar but uncharged side chain. Of the non-polar amino acids, phenylalanine, diphenylalanine, isoleucine, and valine are preferred, and the D-enantiomer is particularly preferred. Of the polar amino acids,
Suitable examples are tyrosine and 4-nitrophenylalanine. Alternatively, when F is absent (see below), E can be a dicarboxylic acid containing up to 6 carbon atoms and lacking the amino group of acidic amino acids. Suitable examples are glutaric acid and succinic acid.

F may not be present (alone or with E), but if present,
Amino acids or analogs with acidic side chains. Aspartic acid is preferred, but glutamic acid is also possible. Like E, F can also be a dicarboxylic acid containing up to 6 carbon atoms and lacking the amino group of acidic amino acids. Examples are glutamic and succinic acids.

Preferably, at least E of residues E and F is present. In particular, it is preferable that both are present.

The amino acids and analogs AF are either the L- or D-enantiomer, but generally the L-form is preferred at all residues. In some cases, it may be advantageous for one or the other of the residues to be in the D-form while the rest are in the L-form. In particular, it may be advantageous for E to be D-glu.

Preferred examples of the peptide “Pep” are listed in Tables 2 and 3 below together with the IC ₅₀ when the C-terminus is not extended. All compounds except those having a succinyl residue at the N-terminus were N-acetylated at the N-terminus.

[0033]

[Table 2] Particularly preferred examples of Pep are shown in Table 3 below together with their IC ₅₀ (μM).

[0034]

[Table 3] ^* Tested only as decapeptide.

In these compounds, Alg = allylglycine. MGly = methylglycine. MVal = methylvaline. Abu = 2-aminobutyric acid. GluS = N-succinylglutamic acid. AsGlu = glutamic acid having an acylsulfonamide at the N-terminus. Cha = β-cyclohexylalanine. Nap = naphthylalanine. AspS = N-succinyl aspartic acid. Nle = norleucine. Dif = 3,3-diphenylalanine. Tha = 2-thienylalanine. FCI = 4-chlorophenylalanine. Phg = phenylglycine. CysMe = S-methylcysteine. Cys (ACS) = Cysteine with C-terminal acylsulfonamide. DHAla = dehydroalanine. Cpc = 1-amino-1-cyclopentanecarboxylic acid. CnAla = cyanoalanine. MGlu = N-methylglutamic acid. Fno = 4-nitrophenylalanine. Gla = γ-carboxyglutamic acid. Dap = β-diaminopropionic acid. Dns = dansyl (5-dimethylamino-1-naphthalenesulfonyl).

Examples of the compounds of the present invention may be effective as inhibitors of NS3 protease at the micromolar or nanomolar level. Preferably, the IC ₅₀ is less than 100 nM, particularly preferably less than 20 nM and optimally less than 5 nM, as measured in the following assay.

According to a second aspect, the present invention relates to any therapeutic use, preferably HCV NS3
There is provided a compound, salt or derivative according to the first aspect of the use for inhibiting proteases and / or for treating or preventing hepatitis C or related conditions. A "related condition" is directly or indirectly caused or can be caused by hepatitis C virus.
In any case, it means a condition associated with HCV.

According to a third aspect, the present invention provides the use of a compound or derivative according to the first aspect in the manufacture of a medicament for treating or preventing hepatitis C or a related condition.

A fourth aspect of the present invention provides a pharmaceutical composition comprising one or more compounds or derivatives according to the first aspect.

The composition may also include pharmaceutically acceptable auxiliaries, such as carriers, buffers, stabilizers, and other excipients. This may further include other therapeutically active agents, especially for use in treating or preventing hepatitis C or related conditions.

The pharmaceutical composition may be in any suitable form, depending on the intended method of administration. The form may be, for example, in tablet, capsule or liquid form for oral administration or in solution or suspension for parenteral administration.

According to a fifth aspect of the present invention there is provided a method of inhibiting HCV NS3 protease activity and / or a method of treating hepatitis C or a related condition, the method comprising the steps of: Or administering to an animal (preferably a mammal) subject a therapeutically or prophylactically effective amount of a composition according to the fourth aspect of the invention or a compound or derivative according to the first aspect. An "effective amount" is one that effects a subject or
It means at least an amount sufficient to cause a change in the condition of the subject.

The dosage rate at which the compound, derivative, or composition is administered will depend on the nature of the subject, the nature and severity of the condition, the manner of administration used, and the like. Appropriate values can be selected by a trained physician. The daily dose of the preferred compound is about 1-100 mg
Would be in the order of The compounds, derivatives, or compositions can be administered alone or in combination with other treatments, either simultaneously or sequentially. Administration can be by any suitable route (oral, intravenous, dermal, subcutaneous, etc.). Intravenous administration is preferred. It can be administered directly at the appropriate site or in a manner that targets a particular site (a type of cell) (appropriate targeting methods are already known).

A sixth aspect of the present invention relates to a method of preparing one or more compounds or derivatives according to the first aspect of the present invention and one or more pharmaceutically acceptable auxiliaries and / or one or more pharmaceutically acceptable auxiliaries. There is provided a method of preparing a pharmaceutical composition comprising the step of mixing with other therapeutic or prophylactic active agents.

According to a seventh aspect of the present invention, there is provided a process for preparing a compound of formula I. These compounds can be made wholly or partially by chemical synthesis, preferably starting from the respective protected amino acid or oligopeptide, and by known peptide synthesis methods.

Embodiments of the Invention Embodiments of the invention are illustrated by way of example only.

EXAMPLES (1) Synthesis One synthesis of the compounds of the present invention is described below. Other compounds can be synthesized by similar methods.

Ac-ASp- (D) Glu-Leu-Ile-Cha-Cys-Pro-C
ha-Asp-Leu-Pro- Tyr-Lys (N ε -Ac) Synthesis of -NH ₂

Solid phase continuous flow Fmoc-polyamide process (Atherton, E. and S
Heppard, R.A. C. , 1989, Solid phase peptide
e synthesis, A practical approach, IRL
Press, Oxford. ). The resin used was modified Rink
Amide linker p-[(R, S) -α- [1- (9H-fluoren-9-yl)
-Methoxyformamide] -2,4-dimethoxybenzyl] -phenoxyacetic acid (
Rink, H .; 1987, Tetrahedron Lett. , 28, 37
87-3789, Bernatowicz, M .; S. Daniels, S .; B
. and Koster, H .; , 1989, Tetrahedron Let
t. , 30, 4645-4667).
All coupling reactions were performed using Fmoc-amino acid / PyBOP / HOBt / DI
Using EA (1: 1: 1: 2) activation with a 5-fold excess of activated amino acids on amino groups not bound to resin for 30 minutes, using double coupling for cysteine residues did. At the end of the assembly, the dried peptide-resin was treated with trifluoroacetic acid / water / triisopropylsilane (92.5: 5: 2.5) for 1.5 hours at room temperature. The resin was filtered, the peptide was precipitated with cold methyl t-Bu ether, and the precipitate was redissolved in 50% water / acetonitrile containing 0.1% TFA and lyophilized.

A homogeneous purification exceeding 98% was carried out using (A) 0.1% trifluoroacetic acid aqueous solution and (B) 0.1% trifluoroacetic acid in acetonitrile as eluents.
Performed by preparative HPLC equipped with a Waters RCM (C-18) column (100 × 25 mm, 15 mm). The gradient used was a constant composition of 40% B for 5 minutes, 40-60% for 20 minutes or more (flow rate 30 ml / min). Fractions were converted to HPL
C (column: Beckman Ultrasphere, C-18, 25 × 4.
6 mm, 5 mm; gradient: 35-65% for 20 minutes, the same eluate as in the fractionation, flow rate 1 m
1 / min), the fractions containing pure material were pooled and lyophilized (yield 5
0%). Mass spectra were obtained on a Perkin-Elmer API-100 spectrometer: MS = 1695.03 (calculated) 1694.6 (found).

(2) Inhibition Assay The ability of the compounds to inhibit NS3 protease was determined by examining the NS3 protease domain and the modified form of the NS4A peptide, Pep 4AK [KKKKSVVIVGRIILS
GR (NH ₂ )] was evaluated using an NS3 / 4A complex. H as a substrate
The substrate peptide 4AB [DEMEECASHLPYK] based on the sequence of the NS4A / NS4B cleavage site of the CV polyprotein was used.

57 μl of 50 mM Hepes (pH 7) with addition of 3 μl of substrate peptide
. 5) 1% CHAPS, 15% glycerol, 10 mM DTT (buffer A)
The cleavage assay was performed in NS4A protein as a protease cofactor
A peptide spanning the central hydrophobic core (residues 21-34), Pep4AK [KK
KGSVVIVGRIILSGR (NH₂)]It was used. 80 μM Pep4
Buffer containing AK was preincubated with 10-200 nM protease for 10 minutes.
And the reaction was started by the addition of the substrate. 6-day data with different substrate concentrations
The velocity parameters were calculated using the data points. Substrate conversion is less than 7%
Incubation time was selected as described above, and the reaction was performed with the addition of 40 μl of 1% TFA.
Therefore, it was stopped. Cleavage of peptide substrates was performed using Merc with autosampler.
It was identified by HPLC using k-Hitachi chromatograph. 80μ
l of the sample was applied to a Lichrospher C18 reverse phase cartridge column (4
× 74 mm, 5 μm, Merck) and fragments were 10-40%
Separation was performed at a flow rate of 2.5 ml / min using a 5% / min gradient of cetonitrile. peak
Is detected by measuring absorbance at 220 nm and tyrosine fluorescence (λ_ex= 260 nm, λ _em = 305 nm). Cleavage of cleavage products with appropriate standards
Quantification was achieved by integration of the chromatogram. Velocity parameters were set to Michaelis-Mainte
Substrate kinetics using Kaleidagraph software, assuming
It was calculated by nonlinear least squares method of initial velocity as a function of degree.

The K ₁ value of the peptide inhibitor was calculated from a substrate titration experiment performed in the presence of increasing amounts of the inhibitor. The experimental data set was fit to Equation 1 using a multi-curve fit macro using Sigmaplot software.

[0054] _{V = (V max S) /} (K m (1 + k i / I) + S) ( Formula 1) or the formula _{2 IC50- (1 + S / K} m) K i ( Equation 2)
The K _i values were derived from the IC 50 values using a 4-parameter logistic function according to

The following table shows the IC ₅₀ values for the various peptides tested in this assay, demonstrating that some optimized compounds of the invention are active at nanomolar or subnanomolar levels. All test compounds, with the exception of compound 26, which has a succinyl residue at the N-terminus, were tested as N-acetyl derivatives.

Some of these compounds are the most potent in vitro inhibitors of HCV protease reported to date. These compounds are non-covalent inhibitors that are reversible and do not contain an electrophilic (“serine capture”) moiety in the molecule. This is appropriate for performing competitive binding assays because these compounds bind to both the S and S 'regions of the enzyme and compete with compounds that bind to either the S or S' region of the enzyme.

[0057]

[Table 4] Abbreviations used in Table I. Abu = aminobutyric acid Cha = β-cyclohexylalanine Hof = homophenylalanine Hyp = hydroxyproline Lys (Ac) or K (Ac) = Nε-acetyl-lysine Nle = norleucine Phg = phenylglycine Sta = statin [(3S, 4S)- 4-Amino-3-hydroxy-6-methylheptanoic acid] Dif = 3,3-diphenylalanine Suc = succinyl N-methylation is indicated by describing (Me) before the three-letter code of amino acids. PYK = proline-tyrosine-lysine (3) Substrate Assay To identify whether an inhibitor molecule is a substrate for the HCV NS3 protease, a modified version of the cleavage assay described above can be used to modify the NS3 protease domain and the modified form as described above. NS4A peptide Pep 4AK [KKKKSV
VIVGRIILSGR (NH ₂ )].

[0058] 1 μM enzyme conjugate was incubated for 16 hours in the presence of 10 μM inhibitor as a candidate substrate peptide. 57 μl of 50 mM Hepes (pH
7.5) Assays were performed in 1% CHAPS, 15% glycerol, 10 mM DTT.

Then, using HPLC, any peptide resulting from the cleavage was separated, and the cleavage product was detected.

A sample was prepared using a Beckman 0.1 equilibrated with 95% solvent A (0.1% TFA in H20) and 5% solvent B (0.1% TFA in acetonitrile).
Analysis was performed by HPLC using a 46 × 25 cm C18 reverse phase column at a flow rate of 1 ml / min. Samples were eluted from this column with a linear gradient of 5% to 90% B for 45 minutes. Peak detection was performed by monitoring the absorbance at 220 nm.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C07K 7/08 A61K 37/02 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, GM, KE, LS, MW, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ, MD, RU) , TJ, TM), AE, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, CA, CH, CN, CR, CU, CZ, DE, DK, D , EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR TT, TZ, UA, UG, US, UZ, VN, YU, ZA, ZW (72) Inventor Bianchi, Elisabetita Italy, I-1995 Rome, Via Saboteno, 31F term (reference) 4C084 AA02 AA07 BA01 BA10 BA23 CA59 DC32 NA01 NA14 ZA751 ZB331 ZB332 ZC202 4H045 AA10 AA30 BA10 DA55 EA29 EA50 FA34 FA44 GA25

Claims (16)

[Claims] 1. A compound of formula (I) or a pharmaceutically acceptable salt or derivative thereof. Pep-A'-B'-C'-D '(I) wherein Formula (I) is described from the left N-terminal to the right C-terminal, and "Pep" binds to HCV NS3 protease. A 'is an optionally substituted proline residue; B' is an amino acid or amino acid analog having a non-polar side chain; C 'is a polar or peptide analog; D 'is leucine, another amino acid or amino acid analog having a non-polar aliphatic side chain, and leucine or other amino acid having a non-polar aliphatic side chain as the N-terminal residue. Alternatively, the bond between Pep and A 'can be substantially cleaved by the HCV NS3 protease. It has.) 2. The compound according to claim 1, wherein the C-terminus is amidated, a pharmaceutically acceptable salt or derivative thereof. 3. A compound according to claim 1 or claim 2, wherein the N-terminus is acylated, or a pharmaceutically acceptable salt or derivative thereof. 4. A tetrapeptide of the formula Pro-B'-C'-Leu, wherein B 'and C' are as defined in claim 1, wherein A'-B'-C'-D 'is A compound according to any one of claims 1 to 3,
Salt or derivative. 5. B ′ is β-cyclohexylalanine, phenylglycine,
The compound, salt or derivative according to any one of the preceding claims, wherein the compound, salt or derivative is selected from homophenylalanine, norleucine, leucine, methionine, norvaline, and β-cyclopropylalanine. 6. The compound, salt or derivative according to claim 5, wherein B ′ is selected from cyclohexylalanine and phenylglycine. 7. C ′ is aspartic acid, glutamic acid, γ-carboxyglutamic acid, glutamine, asparagine, hydroxyproline, N-β-Aloc
The compound, salt or derivative according to any one of the preceding claims, which is selected from diaminobutyric acid, thiazolylalanine, methionine sulfoxide, pyridylalanine and serine. 8. The compound, salt or derivative according to claim 7, wherein C ′ is aspartic acid. 9. The combination of amino acids B′C ′ is Cha-Ser Cha-Asp Nle-Asp Hof-Asp Phg-Asp Cha-Gln Nle-Gln Hof-Gln Cha-Hyp Nle-Hyp Hof-Hyp Nle-Ser. The compound, salt or derivative according to any one of the preceding claims, wherein the compound, salt or derivative is selected from: 10. Pep-OH is capable of binding HCV NS3 protease in the absence of C-terminal residues A'-B'-C'-D 'and has an IC ₅₀ of less than 100 μM in an inhibition assay. A compound, salt or derivative according to any one of the preceding claims. 11. The method according to claim 11, wherein Pep is of the following formula (II): FEDCBA, wherein A is an amino acid or an amino acid analog having an aliphatic side chain of 1 to 6 carbon atoms. B is an amino acid or analog containing a nonpolar, acidic, or polar but uncharged side chain; C is an amino acid or amino acid analog having a nonpolar or acidic side chain; D is E is an amino acid or amino acid analog having a hydrophobic side chain, E may be absent with F, and if present, is an amino acid comprising an acidic, non-polar, or polar, but uncharged side chain Or an amino acid analog or dicarboxylic acid containing up to 6 carbon atoms and lacking the amino group of an acidic amino acid, wherein F may be absent (alone or with E), ,
6. A hexapeptide, pentapeptide, or tetrapeptide having an amino acid or analog having an acidic side chain, or a dicarboxylic acid containing up to 6 carbon atoms). Compound, salt, or derivative. 12. A is selected from cysteine, aminobutyric acid, di- and trifluoroaminobutyric acid, norvaline, allylglycine, and alanine; B is glutamic acid, aspartic acid, glycine, methylglycine, 2-aminobutyric acid, Alanine, isoleucine, valine, leucine, cysteine, naphthylalanine, and β-cyclohexylalanine, wherein C is glutamic acid, aspartic acid, glycine, methylglycine, 2-aminobutyric acid, alanine, isoleucine, valine, leucine, cysteine, Selected from naphthylalanine and β-cyclohexylalanine, wherein D is methionine, isoleucine, leucine, norleucine, valine, methylvaline, phenylglycine, diphenylalanine, tyrosine, thienylala E is selected from glutamic acid, aspartic acid, phenylalanine, diphenylalanine, isoleucine, valine, tyrosine, 4-nitrophenylalanine, glutaric acid, and succinic acid, and F is aspartic acid, glutamic acid. The compound according to claim 11, wherein the compound is selected from:, glutaric acid, and succinic acid. 13. A compound according to any one of the preceding claims for therapeutic use,
Salt or derivative. 14. A pharmaceutical composition comprising a compound, salt or derivative according to any one of the preceding claims and a pharmaceutically acceptable excipient, diluent or carrier. 15. Use of a compound, salt or derivative according to any one of the preceding claims in the manufacture of a medicament for the treatment or prevention of hepatitis C or related conditions. 16. A method for inhibiting HCV NS3 protease activity and / or a method for treating or preventing hepatitis C or a related condition, which is effective in treating or preventing a human or mammal subject suffering from the condition. 13. A method comprising administering a composition according to claim 14 or a compound according to any one of claims 1 to 12.