WO2001064716A1

WO2001064716A1 - Antiviral compounds

Info

Publication number: WO2001064716A1
Application number: PCT/JP2001/001642
Authority: WO
Inventors: Nobutaka Fujii; Hideki Nakashima
Original assignee: Nobutaka Fujii; Hideki Nakashima
Priority date: 2000-03-03
Filing date: 2001-03-02
Publication date: 2001-09-07
Also published as: AU2001236065A1

Abstract

Novel antiviral compounds represented by the general formula (I) which have an excellent antiviral activity and the stability of which is improved in vivo or pharmaceutically acceptable salts thereof and anti-HIV agents containing the same as the active ingredient.

Description

Specification

Technical Field of Antiviral Compounds

INDUSTRIAL APPLICABILITY The present invention has an excellent antiviral activity by acting as an antagonist against a novel antiviral compound, specifically, a glycoprotein, particularly a CXCR4 chemokine receptor, and has improved in vivo stability. It relates to a peptidic compound. Furthermore, the present invention relates to an anti-HIV agent containing the antiviral compound as an active ingredient. Background art

Polypeptides derived from captogae (genus Tachypleus, Limulus, Carcinoscopius), which have strong affinity for endotoxin, have five structural analogs, each consisting of 17 or 18 natural amino acids It is a polypeptide having a cyclic structure, and it has been known that these polypeptides have antibacterial activity (Matsuzaki K., Biochem. Biophys. Acta 1070, 259-264 (1991)).

WO90 / 043474 focuses on the strong endotoxin affinity of the Limulus aegypti polypeptide and discusses its structural transformation and anti-human immunodeficiency virus (HIV) activity, especially HIV A study of the correlation with the receptor describes that a novel polypeptide having high anti-disease activity was obtained.

On the other hand, as a drug target for acquired immunodeficiency syndrome (AIDS) chemotherapeutic agents caused by human immunodeficiency virus (HIV) infection, in addition to reverse transcriptase and HIV protease, The CXCR4 chemokine receptor has attracted attention, and it has become clear that the polypeptide is a peptide antagonist of the CXCR4 chemokine receptor (Tamamura H. et al., Biochem. Biophys. Res. Coraraun. 253, 877-882 (1998)).

However, since the polypeptides disclosed in the prior art are composed of natural amino acids, (i) the terminal amino acid is degraded by endopeptidase, and (ii) the disulfide bond in the polypeptide molecule under reducing conditions. Is cut off, and (iii) poly In addition to the stability problems, such as the turn part of the peptide being susceptible to protease-induced ^^, reduction in the molecular weight of the peptide was required for the purpose of improving activity and reducing toxicity (Arakaki R. et al., J. Virology 73, 2, p. 1719-1723 (1999)). Disclosure of the invention

An object of the present invention is to provide an antiviral compound having a low molecular weight and an improved stability of the polypeptide.

The present inventors have found that the above-mentioned problems can be solved by deamidating the amino acid in the turn portion of the polypeptide, making the disulfide bridge an alkyl or alkylene bridge, or modifying the peptide terminal. . Accordingly, the present invention provides a compound of formula (I):

{Wherein, X is NH ₂ or NH = C (NMe ₂ ) ₂ [where Me is a methyl group];

X ₂ is an amino acid having an aromatic ring;

X ₃ is a single bond or one CR—CH—, wherein is a hydrogen, C Cs alkyl or halogen atom;

X ₄ is NHR ₂ wherein R ₂ is hydrogen or C Cs alkyl; or OH;

X ₅ is one CH ₂ —S—S—CH ₂ —, or C ₄ -C _{8 a} 7 ylene or C ₄ -C ₈ anolekenenylene;

Is an L-amino acid or a D-amino acid selected from Arg, Lys, Orn or other basic amino acids;

Y ₂ is an L-amino acid or a D-amino acid selected from Pro, A la, Va 1 or other aliphatic amino acids; and C it is L-citrulline;

Where X ^ SNH ₂ , X ^ SN a 1, Y—X ₃ — Y ₂ is d Ly s—Pro, X ₄ is OH, and X _{5 is} —CH ₂ —S—S—CH ₂ — Case; and

Is NH ₂ , X ₂ is T rp, — X ₃ — Y ₂ is d Lys — Pro, X ₄ is OH, and X ₅ is one CH ₂ — S— S— CH ₂ — Excluding}

Or a pharmaceutically acceptable salt thereof.

The present invention also relates to an anti-HIV agent containing the ^: virus compound as an active ingredient.

The peptide chain in the novel anti-disease compound of the present invention can be produced by a known method, for example, a solid phase synthesis method. For example, the carboxyl group of N-protected arginine may be bonded directly or, in some cases, via a spacer having a functional group capable of bonding to the carboxyl group or a carboxyl group to an insoluble resin having an amino group, to give each protected amino acid. Are sequentially bonded in accordance with the solid phase synthesis method, and then the protecting group of the insoluble resin and the amino acid is removed to obtain the following formula ():

Xi-Arg- Arg- X ₂ -Cys-Tyr- Arg- Lys- Y factory X ₃ -Y ₂ -Tyr- Arg- Cit- Cys- Arg- Χ ₄ ()

[Wherein the definitions of X ₂ , X ₃ , X ₄ , Y and Y ₂ are the same as those of the above formula (I)]. In this case, the canolepoxyl terminus of the amino acid residue at the C-terminus may be free (X ₄ corresponds to -OH) or converted to an acid amide (X ₄ corresponds to one NH ₂ ) . Further, C ₁ -C ₅ alkyl can be added by a known method (X ₄ corresponds to one NHR ₂ ).

In the solid phase synthesis method, D-amino acids can be used in addition to L-amino acids. The desired D-amino acid can be introduced by using the protected D-amino acid for solid phase synthesis.

To synthesize the polypeptide chain in the compound of the present invention, the carboxyl group of the N-protected arginine at the C-terminal via the amino group or, optionally, the sugar group attached to the carboxyl group of the spacer is bonded. It is an insoluble resin having an amino group, which is possible and can be removed thereafter. Such resins include, for example, aminomethyl resin Fatty acid (aminomethinolelated styrene-dibutylbenzene copolymer), benzhydrylamine resin, methylbenzhydrylamine resin, dimethoxybenzhydrylamine (DMBHA) resin, aminomethylphenoxymethyl resin These derivatives are included. When methylbenzhydrylamine resin, dimethoxybenzhydrylamine (DMBHA) resin, or aminomethylphenoxymethyl resin can be used to give an amide directly by cleavage, aminomethyl resin is preferred in terms of yield . Examples of the spacer having a hydroxyl group and a carpoxyl group capable of binding to a lipoxyl group include, but are not particularly limited to, those capable of converting a carboxyl group of arginine into a lipoxymethyl ester.

The protecting group for the amino group of the amino acid which can be used for the synthesis of the polypeptide chain in the compound of the present invention is, for example, t-butyloxycarbonyl (B oc) or 9-fluoroenylmethyloxycarbonyl. It is. The protecting group for the arginine guanidino group is, for example, tosyl (To s), nitro, 4-methoxy-1,2,3,6-trimethylbenzene snorehoninole (Mtr) or 2,2,5,7,8-pentamethylchroman One is 6-snolehonyl (Pmc). The protecting groups for the mercapto group of cysteine include, for example, benzyl (Bz1), 4-methoxybenzyl (MBz1), 4-methylbenzyl (41-MeBz1), acetamidomethyl (Acm), trityl (Tr t), force including 3-nitro-2-pyridinesulfenyl (Npys), t-butyl (t-Bu), t-butylthio (t-BuS), etc. MBz1, 4-MeBz1 , Trt, Acm and Npys are preferred. Hydroxyl group of tyrosine, for example B z 1, 2, 6- dichloro port base Njiru _{(C 1 2 · B zl)} , may not force \ or protected protected with t-Bu. Protecting groups for the ε-amino group of lysine are, for example, benzyloxycarponyl (Ζ), 2-chlorobenzoyl canoprene (C 1 · Z), Boc or Npys. It is preferable that each protecting group is appropriately selected in consideration of the peptide synthesis conditions and the like.

The binding of the protected amino acid can be carried out by a conventional condensation method such as DCC (diclohexyl canoleopreimide) method, DIP CDI (disopropyl carpoimide) method, active ester method, mixed or symmetric acid. Anhydrous method, Carboerdiimidazole method, DC C-HOB t (1-hydroxybenzotriazole) method, Diphenyl phosphoryla Although it can be carried out by a zid method or the like, a DCC method, a DCC-HOBt method, a DIPCDI method, and a symmetric acid anhydride method are preferred.

In the antiviral compound of the present invention represented by the formula (I), X is NH ₂ or NH = C. (NMe ₂ ) ₂ [wherein Me is a methyl group]. X in the formula (I) is preferably NH = C (NMe ₂ ) ₂ because the peptide moiety is less likely to be degraded by endopeptidase. When a compound in which X is NH = C (NMe ₂ ) ₂ is desired, for example, the α-amino group of arginine at the N-terminal is converted into ΝΗ = C (NM e ₂ ) _2, and the converted arginine is converted to arginine. It can be obtained by using it for peptide synthesis.

From the same viewpoint, X ₄ in formula (I), wherein 1 ₂ is a 〇 ₁ ～〇 ₅ Arukiru] NHR ₂ preferably is.

In antiviral compound of formula (I) in the present invention, as the § amino acid having an aromatic ring of X ₂ are also encompassed Amino acids having an aromatic ring other than the essential Amino acids such Toributofan, antiviral of the compound For reasons of increased activity and / or reduced toxicity, equation (II): 6

(Π)

NH-CH-CO

[Where x ₆ is

Amino acid having an aromatic ring represented by the following formula:

In the present invention, X ₂ can be tributophan (X ₆ is [TRP]), but tryptophan 1H-indole (TRP) can be replaced with Na 1, Cys (Bzl), Cys (Ad) or T rp (M ts) the substituted o; - by using amino acid peptidyl de synthesis, it is possible to obtain a desired X _2.

Formula (I) wherein —X ₃ —Y ₂ , preferably dLys-Pro or dLys—CH = CH—Ala, where dLys represents D—lysine. However, dLys-CH = CH-A1a is more preferable because the stability of the compound is improved by making the turn portion of the i3 sheet structure of the compound of the present invention less susceptible to peptidase. .

In the compound of the present invention, when a compound in which Yi—X ₃ —Y ₂ which is the turn part of the three-sheet structure is d Lys—CH = CH—A 1 a is desired, the amino acid is used as a chiral pool, By using a unique EAD I ((E) -alkene dipeptide isostere) synthesis method that utilizes the stereospecific anti-SN 2 'reaction with organocopper reagents, D-Lys-A1a type EAD I, Can obtain a compound into which Fmo c— DLy s (CIZ) —Ψ [(E) CH = CH] -A 1 a is introduced (Tamamura H., J. Chem. Soc, Perlkin Trans I 1999, 2983- 2996).

In the compound of formula (I), a disulfide bond (X _{5 is} —CH ₂ —S—S—CH ₂ —) with cysteines at the 4- and 13-positions forms an S—S bridge, for example, via a mercapto group. Can be introduced. In this case, the mercapto group is protected with a protecting group t-BuS, and t-BuS is deprotected and oxidized to form a disulfide bond. In this case, a known method can be used for the oxidation treatment, but usually an oxidizing agent such as oxygen in the air or a ferricyanate (eg, potassium ferricyanide) is used.

In order to improve the stability of the compound of the present invention as a non-peptide, X ₅ in the formula (I) is represented by the following general formula:

It is preferably C ₄ -C ₈ alkylene or C ₄ -C ₈ alkenylene selected from the group consisting of pentamethylene, cis-3-pentenylene, trans-2-pentenylene or hexamethylene. preferable.

The X ₅ to a C ₄ -C ₈ alkylene or C ₄ -C ₈ alkenylene, constructing the peptide chain using instead Arirugurishin (Allylglycine) or homo-allyl glycine (Horaoallylglycine) derivatives of Shisuti emissions derivatives, R u Alkene or alkane bridges can be formed by a catalytic RCM (ring closing olefin metathesis) reaction. The derivative obtained in this way was subjected to conformation analysis by CD analysis, and it was shown that the compound derived from arylglycine and homoarylglycine had a β-sheet structure similar to that of the original peptide.

The compounds obtained as described above are subjected to extraction, recrystallization, various chromatographies (eg, gel filtration, ion exchange, partitioning, adsorption, reverse phase), electrophoresis, countercurrent partitioning, and other techniques. Reversed phase high performance liquid chromatography, which can be isolated and purified by methods known for peptides, is preferred.

The compounds represented by the formula (I) of the present invention are exemplified in Table 1 below. The compounds Τ134 and Τ140 were prepared according to the prior art described above (Tamamura H. et al., Biochem. Biophys. Res. Coraraun. 253, 877-882 (1998); Arakaki R. et al. al., J. Virology 73, 2, p. 1719-1723 (1999)).

Examples of specific structures of the peptide compounds of the present invention shown in Table 1 are shown below.

Replacement form (Rule 2

Replacement form (Rule 26) m) ^ Ύ m

l / Zl

9UP9 / W OAV 12/2

Replacement form (Rule 26) (u) m ^

Odf / e :) d 9Ϊ.1-9 / Ϊ0 OAV 13 The physical properties of the peptide compounds of the present invention are shown in Table 24 below.

3 Sμα 0μ Dimension 1 Jzq 1φΛβ0s βφseΛz

Replacement form (Rule 26) M 3

MS Characterization of T (E) -140 and Its Related Peptides

Compound X 6 X ₄ Formula Ion sprav mass (reconstructed)

Four

Physicochemical Constants of T (E) -140 and Its Related Peptides

Compound XX, 6 X ₄ [a] _D (Cone, in H ₂₀ , temp)

T134 ^ N ^ NNNNN ^ NN-N 0 (C = 0.10, 16 ° C) T140 ^^ HHHHHHHHH HH

^ ^^ 22222222222 ",-0.01 (C = 0.24, 27.C)

TMG-140-NH2 -7.2 (C = 0.55, 25 ° C)

T (E) -134-OH ^-M M

¾ e e -17.1 (C = 0.31,27 ° C)

T (E) -140-OH -11.5 (C = 0.52, 25 ° C)

T (E) -134-NH2 T CC -14.6 (C = 0.27,37 ° C)

T (E) -140-NH2 PSTT TNNNNNN N -8.1 (C = 0.37, 36 ° C)

T (E) -141-NH2

T (E) -142-NH2 -0.9 (C = 0.22, 28 ° C)

-12.0 (C = 0.25, 38 ° C)

T (E) -143-NH2 + 2.5 (C = 0.16, 29 ° C)

TM (E) -134-OH OOOONOO ONNNNON

TM (E) -140-OH -3.3 (C = 0.57, 26 ° C)

HHiHH H

TMG (E) -140-OH -6.5 (C = 0.29, 27 ° C) des-Argl-TMG (E)--2.6 (C = 0.39, 19 ° C)

140-OH -24.2 (C = 0.29,22 ° C)

The antiviral compound of the present invention exemplified in this way has a high antiviral activity and a low toxicity equivalent to or higher than that of a known compound, and has improved stability in a living body. In the compounds of the present invention, modification of the distal portion of the X and X _4, non-peptide of the turn portion of the 0 sheet over preparative, as well as the alkylene reduction or alkenylene of X _5, the compounds in vivo metabolic effects such as proteases In vivo stability is imparted because the peptide compound is less susceptible to aging and because the cyclic structure of the peptide compound is prevented from being linearized due to dissociation of the Z or SS bridge.

The drug containing the antiviral compound represented by the formula (I) of the present invention as an active ingredient contains the compound represented by the formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient, and is used for virus infection or virus. It can be administered orally or parenterally for the disease. The drug of the present invention can be formulated into various dosage forms, such as tablets, granules, capsules, powders, suppositories, pessaries, ointments, creams, aerosols, liquids, such as liquids for internal use, injections, and infusions. Can be administered. Depending on the method and mode of administration of the drug, various pharmaceutically acceptable additives may be used, for example, excipients, binders, disintegrants, buffers, dissolution aids, isotonic Agents, P-preservatives, antioxidants, pH regulators, suspending agents, diluents and the like.

The daily dose of the drug of the present invention varies depending on the method of administration, the dosage form and the degree of symptoms. For example, when administered orally as a tablet, the antiviral compound of the present invention or a pharmaceutically acceptable salt thereof is administered in a range from 0.1 mg / kg to human LOOOOmg / kg human body weight. In the case of parenteral administration as an injection, it is administered in the range of 0.1 to 100 mg / kg human body weight. The daily dose of the agent of the present invention can be administered one or more times, but more generally, once to three times a day, a unit dose is orally or parenterally administered. Is preferred. BEST MODE FOR CARRYING OUT THE INVENTION

The present invention is not limited to the following examples. 17 Example

Reference Example 1 Production of T134

The following array: '

A polypeptide having NH ₂ -Arg-Arg-Trp-Cis-Tyr-Arg-Lys-Dlys-Pro-Tyr-Arg-Cit-Cys-Arg-C00H was synthesized as follows.

People are aminomethyl resin and _{Fmo c-DMBHA-CH 2 CH} 2 COOH to the solid-phase synthesis column, 2 Tokitoichijimigohan jfe was performed by DI PCD I -HOB t method. After the completion of the condensation reaction, force coupling was performed using anhydrous acetic anhydride to protect free amino groups (DMBHA resin). After removing the Fmo c group of the DMBHA resin thus prepared with 20% piperidine ZDMF, 2.5 eq of Fmo c—Ar g (M tr) —OH was added to the DMBHA resin, and DIP CD I— The condensation reaction was performed by the HOBT method. The degree of progress of the condensation reaction was measured by a ninhydrin test.

After removing the Fmo c group of the arginine-introduced DMBHA resin thus prepared with 20% piperidine / DMF, 2.5 eq of Fmo c—Cys (MB zl) —OH was added to the DMBHA resin, and DI PCD I —Condensation reaction was performed by the HOB t method. The degree of progress of the condensation reaction was similarly measured by a ninhydrin test. In the same manner C- terminal sequentially protecting group-protected Amino acid according to the sequence from Amino acid were introduced into DMBHA resin to obtain a protecting group-protected peptide first resin _D then the coercive Mamorumoto protected peptide first resin The Fmoc group is removed by treatment with 20% piperidine ZDMF, and 1 M TMSOTf-thioanisole / TFA per 10 Omg of the resin (m-cresol (100 eq), ethanedithiol (300 eq) The resin was filtered from the reaction mixture to obtain an uncyclized T134 polypeptide, which was then oxidized with air to carry out a cyclization reaction. T 134 polypeptide was obtained Reference Example 2 Production of T 140

The following array: 18

_{NH 2 -Arg-Arg- [Nal]} -Cis- Tyr- Arg- Lys- Dlys- Pro-Tyr- Arg- Cit- Cys- Arg- upon synthesizing a polypeptide having a COOH, 3-position of tryptophan (TRP) Was replaced with L-1- (2-naphthyl) arayun [Na1], and the same operation as in Reference Example 1 was repeated to obtain a T140 polypeptide. Example 1 Introduction of Alkin Bridge to Peptidic Compound

According to the following reaction scheme 1, Fmoc-L-homoallylglycine was synthesized. Reaction Scheme 1

-Serine

Fmoc-Le Hag-OH

Using the obtained Fmoc-L-homoallylglycine, a peptide chain is constructed by a Ru-catalyzed RCM reaction according to the following reaction scheme 2, and hexamethylene is introduced to introduce the cyclic peptide of the present invention. The active compounds TM14Ο, ΗΗ (E), TM140, HH (Z) and TM140, HH (Red) were synthesized.

* Replacement form (Rule 26) 19 Reaction Scheme 2

Similarly, TM140, AH (E), TMl40, AH (Z), TM140, AA (E), TMl40, HA (E), TMl40, HA (Z), TMl40, AH (Red ) And TMl40, AA (Red) were synthesized and CD analysis was performed with T140 synthesized in Reference Example 2. The results showed that the compound of the present invention had a sheet structure similar to that of T140 (FIG. 1). Replacement form (Rule 26) 20 Comparative test 1 CXCR4-chemokine receptor antagonist activity test To evaluate the antagoest activity of each test compound on CXCR4-chemokine receptor, CHO (Chinese Hamster Ovarian) cells overexpressing CXCR4-chemokine receptor In contrast, the ability of each test drug to suppress the increase in intracellular calcium ion concentration induced by human Stromal Cell-derived Factor (SDF), an endogenous ligand of CXCR4-chemokine receptor, was evaluated.

In order to analyze the signal transduction ability by the ligand, measurement of intracellular C a ²⁺ ions was performed. A 1 OmM DMSO solution of each test substance was prepared in advance and diluted with a 20 mM Hepes-containing Hanks solution to an appropriate concentration (final concentration: 0.01 to 30 µΜ) immediately before measurement.

0 Each 3 X 1 cells before 24 or 48 hours of measurement ⁴ or 1. 5 X 1 0 ⁴ cells Zl 5 mu L wells concentration in 96-well microtiter plate (C oster 3 6 3 5 or MCON 3 2 96 ). Immediately before measuring the intracellular Ca ²⁺ concentration, load 5 μΜ Fura 2—AM for 1 to 2 hours in the dark, and wash twice with 100 μl of Hanks solution containing 2 OmM Hepes, then remove the spectrometer. Measure the ratio of 340 nm / 380 nm using ter (FDS S-4000 or FDS S-2000, Hamamatsu Phototus Co., Ltd.). 2 For cells filled with 100 μL / we 11 of Hanks solution containing OmM Hepes, 30 L of test compound solution (final concentration: 10 μM) was added to the test compound 30 seconds after the start of measurement. After one minute, add 40 μL (3 OmM) of the ligand (hSDF) solution, and then monitor the intracellular Ca ²⁺ concentration for another 4 minutes. The inhibition rate of each test compound with respect to the change in Ca ²⁺ concentration in the Vehic 1e-added group is calculated, and the compound concentration at which ₅₀ % inhibition is exhibited is defined as IC ₅₀ . Table 5 shows the results.

Replacement form (Rule 26) twenty one

Table 5

Ango gonist activity

Compound IC ₅₀ (iiM) [CXCR4 / SDF]

T140 G 3

TM140, AH (Z) G 3

TM140, AH (Red.) 3

TM140, HA (E) 1

TM140, HH (Red) 0.3-1

The results of the antagonist activity test in Table 5 show that the antiviral compound of the present invention also acts as an antagonist on the CXCR4 chemokine receptor, similarly to the known compound T140. Example 2 Introduction of (E) ethylene dipeptide (d Lys— CH = CH—A 1 a) into the turn part of the e-sheet in a peptide compound

According to the following reaction scheme 3, Fmoc-DLy s (CIZ) was synthesized using an EAD I ((E) -alkene dipeptide isostere) synthesis method utilizing a stereospecific anti-SN 2 'reaction with an organocopper reagent. (E) CH = CH] —A 1a was synthesized.

22 Reaction Scheme 3

r

Fmoc-DLys (CIZ) -M ^ [(E) CH = CH; i-LAIa Using the obtained Fmoc-DLys (CIZ) Ψ [(E) CH = CH] —A1a, Reaction Scheme 4 was performed to obtain (E) a compound of the present invention into which an ethylene dipeptide was introduced.

Replacement Paper Rules 26 >> 22/1 Reaction Scheme 4

Pmc Ptnc Trt ¾u Pmc Boc Boc ½u Pijic Trt Pmc

Fmoc-Α! ¾— Aig- Y- C s— T r— Arg—S sD “S s-Pra-T r—Arg- Cit-C — Arg-r _{PAL Resin-} cresol, EDT

T (E) -140 Derivatives

Replacement sheet (Rule 26) 23 Example 3 Production of T134 Derivative and Τ140 Derivative

Introduction of the alkane or alkene bridge of Example 1 and / or introduction of the (Ε) ethylene dipeptide (d Lys— CH = CH—A 1 a) of Example 2 was performed using the m (E) ethylene dipeptide (d Lys The compounds of the invention described in Tables 2 to 4 were prepared using the introduction of s-CH = CH-A1a). Comparative test 2 Anti-til V activity test

The antiviral activity against HIV of the compounds prepared in Examples 1 to 3 was determined as follows according to the MTT method (Pauwels et al., J. Virol. Methods, 20; 309-321 (1988)). .

In a 96-well microtiter plate, HIV-infected MT-4 cells (2.5 10 ⁴ cells / well, multiplicity of infection [MO1: 0.01]) were added with various concentrations of the test substance immediately after infection. After 5 days of culture at 37 ° C for at C0 ₂ incubator, it was measured number of remaining cells by MT T method. Antiviral activity, cell failure 50% inhibition concentration by HI V infection: expressed by _{(EC 5. 50% Effective Concentration} ). On the other hand, in order to examine the cytotoxicity of the test compound to MT-4 cells, non-virus-infected cells were similarly cultured with various concentrations of the test substance. Cytotoxicity 50% cytotoxicity concentration by the test substance: expressed by _{(CC 5. 50% Cytotoxic Concentration} ). The effective coefficient (SI: Selectivity Index) was calculated as CC ₅₀ ZE C ₅₀ .

As test substances, the compounds prepared in Examples 1 to 3 as compounds of the present invention, T22 further known as a chemokine antagonist, T134 and T140 prepared in Test Examples 1 and 2, and Azidothymidine (AZT), which is currently used to treat HIV, was used. The results of the anti-HIV activity test are shown in Table 6 below. 24 Table 6

Anti-HIV activity measurement test

Ishi 5 h IJIM) ji so nM bX shi 50 / shi

T22 43.6 Nd

T134 15.1 Nd

T140 c 0.3 66 600> 222000

TMG140-NH ₂ 2.4 59500 24887

T (E) l34-OH 1.3 1.3 1000> 77692

T (E) 140-OH 1.9 269 400 138462

T (E) 134-NH ₂ 2.9 58 200 19901

T (E) 140-NH ₂ 1.9 57500 30775

T (E) 141-NH ₂ 44.8 58500 1306

T (E) 142-NH ₂ 16.9 56500 3339

T (E) 143-NH2 1.4 11800 8284

TM (E) 134-0H 15.7 55800 3551 M (E) 140 ~ OH 11.5 261600 22763

AZT 1.1 45700 35369 The results of the anti-HIV activity measurement test shown in Table 6 showed that the antiviral compound of the present invention had excellent antiviral activity similarly to known anti-HIV agents. Furthermore, since the compound of the present invention has low cytotoxicity and improved in vivo stability, it is expected to be an excellent anti-HIV agent. Industrial applicability

According to the present invention, a novel antiviral compound having excellent antiviral activity against human immunodeficiency virus (HIV) and improved in vivo stability or a pharmaceutically acceptable salt thereof, and An anti-HIV agent containing the active ingredient as an active ingredient is provided. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a graph of the CD analysis pattern of the compound of the present invention, showing the CD spectrum of the TM140 derivative from 80 to 260 nm.

Replacement sheet (Rule 26)

Claims

25 Claims

1. Formula (I):

{Where X is NH ₂ or NH = C (NMe ₂ ) ₂ [where Me is a methyl group];

X ₂ is an amino acid having an aromatic ring;

X ₃ is a single bond or one CI ^ CH—, wherein 1 ^ is hydrogen, C ₁ -C ₅ alkyl or a halogen atom;

X ₄ is NHR ₂ wherein R ₂ is hydrogen or C Cs alkyl; or OH;

¹ X ₅ is one CH ₂ —S—S—CH ₂ —, or C ₄ -C ₈ alkylene or C ₄ -C ₈ alkenylene;

Is an L_amino acid or a D-amino acid selected from Arg, Lys, Orn or other basic amino acids;

Y ₂ is an L-amino acid L-amino acid or a D-amino acid selected from Pro, A la, Va 1 or other aliphatic amino acids; and

C i t is L-chitonorelin;

Where X ^ SNH ₂ , X ₂ is Na 1, Y — X ₃ — Y ₂ is d Ly s—Pro, X ₄ is OH, and X _{5 is} —CH ₂ —S—S—CH ₂ — Case; and

Is NH ₂ , ₂ is rp, — X ₃ — Y ₂ is d Lys — Pro, X ₄ is OH, and X ₅ is one CH ₂ — S— S— CH ₂ —

Or a pharmaceutically acceptable salt thereof.

2. X ₂ is the formula (Π):

I (ID

NH-CH-CO 26

[Where X ₆ is

Selected from the group consisting of the following]: the anti-viral compound or a pharmaceutically acceptable salt thereof.

3. The method according to claim 1, wherein Y—X ₃ —Y ₂ is d Lys—Pro or d Lys—CH = CH—A1a, wherein dLys represents D—lysine. Or a pharmaceutically acceptable salt thereof.

4. X ₅ has the general formula:

Selected from the group consisting of the C ₄ -C ₈ alkylene or C ₄ -C ₈ 7 Luque - Len der Ru, antiviral compounds or acceptable salts their pharmaceutically according to claim 1.

5. is a _{_{NH = C (NMe 2) 2}} , may be antiviral compounds, or their pharmaceutically acceptable according to any one of claims 1 to 5 salt.

6. An anti-HIV agent comprising the antiviral compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof as an active ingredient.