JPH07215940A - Compound having antiviral activity - Google Patents

Abstract

PURPOSE:To obtain a method for the determination of N-myristoyl transferase activity useful for the investigation of the biological significance of myristoylation and obtain a compound having antiviral activity against virus, especially retrovirus, particularly the causal virus (HIV-I) of immunodeficiency syndrome (AIDS). CONSTITUTION:This compound is expressed by formula Z-Y-(CH2)xCO0R (Z is a pyridyl, a pyrimidyl, a triazolyl or a tetrazolyl; Y is oxygen or sulfur atom; R is H or a 1-8C alkyl; x is 7-11) and usable as fatty acid analogs and pharmaceuticals.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a method for measuring N-myristoyl transferase activity using a novel fatty acid analog, a virus and a retrovirus, among which immunodeficiency syndrome (AIDS).
To a compound having antiviral activity against the causative virus (HIV-I).

[0002]

2. Description of the Related Art It is generally known that the amino terminus of certain proteins is blocked by an acetyl group, a pyroglutamyl group, and a formyl group. Myristic acid, a long-chain fatty acid with 14 carbon chains, is covalently bound to the amino terminus of the protein via an acid amide bond.
In 1982, in the process of determining the entire primary structure of AMP-dependent protein kinase, Shoji et al. (Proc. Natl. Ac.
ad. Sci. , U. S. A. , 79, 6128 (19
82)), the identification of amino-terminal myristoyl groups of various proteins has been made.

Amino-terminal myristoylated proteins can be roughly classified into cell-derived proteins and virus-derived proteins. The former is a phosphoprotein phosphatase such as cAMP-dependent protein kinase, calcineurin (calcineuri).
n) B, reduced nicotinamide-dinucleotide phosphate (NADH) -cytochrome b5 reductase, BC
Intracellular protein of 3 H1 muscle cell line, pp60
^There are p36K substrate protein of ^v-src (src-gene gene product, tyrosine kinase), intracellular protein in macrophages, insulin receptor, vinculin and the like.

The latter virus constituent proteins whose myo-terminals are myristoylated include Pr65 ^gag core protein of Lauscher-Moroni murine leukemia virus, gag-onc protein of FeSV, and moroni-.
Murine leukemia virus tyrosine protein kinase,
Rous sarcoma virus (RSV) pp60 ^v-src , adult T-cell leukemia (ATL) virus (HTLV-I) p
19 ^gag protein, p17 ^{ga g} protein, polyomavirus and outside the SV40 capsid protein VP cause viral immune deficiency syndrome (AIDS) (HIV-I)
2. Mason-Pfize
r) A monkey virus core protein, a hepatitis B virus (HBV) pre-S1 core protein, and a poliovirus core protein VP4 are known. A myristoyl group was found in the α-subunit of the human oncogene (src-gene) product pp60 ^c-src and a guanosine tripophosphate (GTP) -binding protein, and a myristoyl group was also suggested to be bound to immunoglobulin. .
As described above, myristoylation of various intracellular proteins, viral constituent proteins, or oncogene products has been found.

The enzyme that catalyzes the amino-terminal myristoylation of proteins is yeast (DA Towler et al., J. Bi.
ol. Chem. , 263, 1784 (1988)) or animal tissue (CJ Glover et al., Bioch.
em. J. , 250, 485 (1988)) and partially purified, but its enzymatic chemical properties are not clear. It is known that this enzyme catalyzes the transfer of a myristoyl group between myristoyl coenzyme (CoA) as a myristoyl donor and an N-terminal glycyl peptide as an acceptor.

The activity of myristoylase is actually carried out as follows. Radioisotope labeled myristoyl donor ie [ ³ H] -myristoyl-Co
A or a myristoyl group receptor, that is, a glycyl peptide is labeled, and a labeled amino-terminal myristoylated peptide generated by an enzymatic reaction is analyzed by high performance liquid chromatography (HPLC) to obtain a method widely used. There is.

Relatively short synthetic N-terminal glycyl peptides that can be acceptors of myristoylase are described in US Pat. Nos. 4,740,588 and 4,778,878. Examples of such peptides include:
Gly-Asn-Ala-Ala-Ala-Ala-A
rg-Arg and Gly-Asn-Ala-Ala-
Ser-Tyr-Arg-Arg.

On the other hand, various biological significances of myristoylation have been discussed. For example, RSV pp60 ^v-src
, The inhibition of myristoylation loses the ability to cause cell phenotypic changes (D. Pellman et al., Nat.
ure, 314, 374 (1985)), and in the poliovirus core protein VP4, myristoylation plays an important role for the virus to retain the capsid structure or in the process of sending self RNA to the host cell. Has been reported (M. Chow
Et al., Nature, 327, 482 (1987)).

S. P. Adams et al. Have oxy or thio fatty acids with carbon chains 13 or 14 in which the 4th to 13th carbons have been replaced by oxygen or sulfur (US Pat.
94), or an azido-substituted oxy or thio fatty acid having a carbon chain of 9 to 13 having an azide, tetrazoyl or triazolyl group at the ω position (US Pat. No. 596,183).
No.), the CoA ester serves as a substrate for myristoylase, and that these fatty acid analogs inhibit the virus in a mammalian host infected with the virus.

[0010]

As described above, as the biological significance of myristoylation and the importance of the antiviral action by inhibiting myristoylation become clear, a simple method for measuring the activity of myristoylation enzyme and a novel method There is a demand for the development of a myristoylation-inhibiting compound.

[0011]

SUMMARY OF THE INVENTION Therefore, the present invention was made by studying various fatty acid analogs and arrived at the present invention. The present invention provides fatty acid analog substrates that are active against enzymes that catalyze protein amino-terminal myristoylation. These compounds, in the fatty acid skeleton, pyridyl group, pyrimidyl group, triazolyl group, tetrazolyl group,
Contains oxygen or sulfur atoms.

Preferred fatty acid analogs have the general formula: Z-Y- in _{(CH 2) x COOR (I} ) formula, Z is a pyridyl group, a pyrimidyl group, a triazolyl group or tetrazolyl group, Y is an oxygen atom or a sulfur atom, R represents hydrogen or C1-8 Alkyl group and x show 7-11. More preferably, in the formula, Z is a pyridyl group or a pyrimidyl group, Y is a sulfur atom, R is H, and x is
Indicates 8 to 10.

[0013] Conventionally, the activity of myristoylase is measured by a radioisotope-labeled myristoyl donor, namely [
³ H] -myristoyl-CoA, or a myristoyl group receptor, that is, a glycyl peptide is labeled, and the labeled amino-terminal myristoylated peptide generated by the enzymatic reaction is separated by high performance liquid chromatography (HPLC), and online emission is performed. Noh detectors are used for evaluation. Each of these novel compounds of the present invention is 25
It is a characteristic compound having an absorption maximum at 0, 300 or 316 nm. By utilizing these absorption characteristics,
Non-radioactive measurement of myristoylase activity has become possible.

In the presence of the compound of the present invention, human T-cells (CEM) are first infected with the causative virus (HIV-I) of immunodeficiency syndrome (AIDS), and the infectious daughter virus is infected with virus (TCID50, cell culture). It was found that the compound of the present invention inhibits virus production when the dose of the virus that infects 50% of the cells in () is used as an index.
Furthermore, the compound of the present invention was used as an HIV-I sustained production strain (CE
It was found that the compound of the present invention inhibits virus production even when it is acted on M / LAV-I).

This is because the novel compound of the present invention inhibits the action of myristoyltransferase, or exhibits a hydrophobicity different from that of myristoylated protein when incorporated into a protein by myristoyltransferase. It is meant to change the cell trait, which is discussed as the biological significance of myristoylation, to retain the capsid structure of the virus in the core protein, or to change the process of sending the viral RNA into the host cell.
That is, the novel fatty acid analog of the present invention provides a useful method for assaying N-myristoyl transferase activity for the search for the biological significance of myristoylation, and a virus,
It is useful as a compound having an antiviral activity against a retrovirus, in particular, an immunodeficiency syndrome (AIDS) causative virus (HIV-I).

According to the present invention, a simple method for measuring N-myristoyl transferase activity using a novel fatty acid analog, and further an antivirus against a virus, a retrovirus, and particularly a virus causing the immunodeficiency syndrome (AIDS) (HIV-I). Compounds having activity are provided.

More specifically, the following fatty acid analogs are provided. 9- (2-pyridylthio) -nonanoic acid (1) 10- (2-pyridylthio) -decanoic acid (2) 11- (2-pyridylthio) -undecanoic acid (3) 9- (4-pyridylthio) -nonanoic acid ( 4) 10- (4-pyridylthio) -decanoic acid (5) 11- (4-pyridylthio) -undecanoic acid (6) 9- (2-pyrimidylthio) -nonanoic acid (7) 10- (2-pyrimidylthio) -decane Acid (8) 11- (2-pyrimidylthio) -undecanoic acid (9) The fatty acid analog of the present invention is not limited to these.

The biologically active fatty acid analogs of the present invention can be prepared by various synthetic routes. For example, an ω-alkyl bromidecarboxylic acid derivative having a required chain length can be prepared, for example, from N, N-dimethylformamide (DM
It can be synthesized by reacting with a thiol compound in an organic solvent such as F) or in the presence of anhydrous potassium carbonate at room temperature, but other equivalent known methods can also be used.

[0019]

INDUSTRIAL APPLICABILITY According to the present invention, simple N-myristoyl transferase activity measurement, and further, inhibition of infection of virus into host cells and inhibition of growth of virus-infected cells were possible.

The following examples illustrate the invention but do not limit it.

Example 1 Preparation of 9- (2-pyridylthio) -nonanoic acid (1) 1 g of 9-nonanoic acid bromide, 0.5 of 2-mercaptopyridine
A mixture of g, 1.2 g of potassium carbonate and 8 ml of N, N-dimethylformamide (DMF) is stirred at room temperature overnight.
After adding 10 ml of water to the reaction solution, it is extracted with diethyl ether. After acidifying the aqueous layer with citric acid, it is extracted with ethyl acetate. The organic layer was washed with water, dried over anhydrous sodium sulfate, and the organic solvent was distilled off under reduced pressure to obtain 0.81 g of 9- (2-pyridylthio) -nonanoic acid. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
_{0-1.51 (m, 10H, SCH 2} CH 2 (CH 2)
₅ ), 1.59 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.19 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.10 (t, 2H, J = 7.
3 Hz, SCH ₂ ), 7.11 (dd, 1H, J = 6.
9 Hz, J = 7.6 Hz, pyridyl-H-5), 7.2
7 (d, 1H, J = 8.0 Hz, pyridyl-H-3),
7.63 (dd, 1H, J = 7.6Hz, J = 8.0H
z, pyridyl-H-4), 8.44 (d, 1H, J =
6.9 Hz, pyridyl-H-6), 11.95 (br
s, 1H, COOH) UV: λmax (nm) = 25
0,316

Example 2 Production of 10- (2-pyridylthio) -decanoic acid (2) In the same manner as in Example 1, 10- (2-pyridylthio) was obtained from 1 g of 10-bromodecanoic acid and 0.44 g of 2-mercaptopyridine.
-0.82 g of decanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
1-1.51 (m, 12H, SCH ₂ CH ₂ ( CH ₂ )
₆ ), 1.58 (quint, 2H, J = 7.2 Hz,
SCH ₂ CH ₂ ) 2.22 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.11 (t, 2H, J = 7.
2 Hz, SCH ₂ ), 7.09 (dd, 1H, J = 7.
0 Hz, J = 7.6 Hz, pyridyl-H-5), 7.2
5 (d, 1H, J = 7.9 Hz, pyridyl-H-3),
7.60 (dd, 1H, J = 7.6Hz, J = 7.9H
z, pyridyl-H-4), 8.42 (d, 1H, J =
7.0 Hz, Pyridyl-H-6), 11.95 (br
s, 1H, COOH) UV: λmax (nm) = 25
0,316

Example 3 Preparation of 11- (2-pyridylthio) -undecanoic acid (3) In the same manner as in Example 1, 11- (2-pyridylthio)-was prepared from 1 g of 11-undecanoic acid bromide and 0.42 g of 2-mercaptopyridine. 0.86 g of undecanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
1-1.52 (m, 14H, SCH ₂ CH ₂ ( CH ₂ )
₇ ), 1.59 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.18 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.11 (t, 2H, J = 7.
3 Hz, SCH ₂ ), 7.09 (dd, 1H, J = 6.
9 Hz, J = 7.6 Hz, pyridyl-H-5), 7.2
5 (d, 1H, J = 8.2 Hz, pyridyl-H-3),
7.62 (dd, 1H, J = 7.6Hz, J = 8.2H
z, pyridyl-H-4), 8.42 (d, 1H, J =
6.9 Hz, pyridyl-H-6), 11.96 (br
s, 1H, COOH) UV: λmax (nm) = 25
0,316

Example 4 Preparation of 9- (4-pyridylthio) -nonanoic acid (4) In the same manner as in Example 1, 9- (4-pyridylthio)-was prepared from 1 g of 9-nonanoic acid bromide and 0.47 g of 4-mercaptopyridine. 0.87 g of nonanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
1-1.52 (m, 10H, SCH ₂ CH ₂ ( CH ₂ )
₅ ), 1.68 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ) 2.22 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.08 (t, 2H, J = 7.
3 Hz, SCH ₂ ), 7.28 (d, 2H, J = 6.4)
Hz, pyridyl-H-3,5), 8.40 (d, 2H,
J = 6.4 Hz, Pyridyl-H-2,6), 11.99
(Br s, 1H, COOH) UV: λmax (nm)
= 230,300

Example 5 Preparation of 10- (4-pyridylthio) -decanoic acid (5) In the same manner as in Example 1, 10- (4-pyridylthio) was obtained from 1 g of 10-bromodecanoic acid and 0.44 g of 4-mercaptopyridine.
-0.89 g of decanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
_{2-1.51 (m, 12H, SCH 2} CH 2 (CH 2)
₆ ), 1.65 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.18 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.04 (t, 2H, J = 7.
3 Hz, SCH ₂ ), 7.25 (d, 2H, J = 6.4)
Hz, pyridyl-H-3,5), 8.36 (d, 2H,
J = 6.4 Hz, Pyridyl-H-2,6), 11.98.
(Br s, 1H, COOH) UV: λmax (nm)
= 230,300

Example 6 Preparation of 11- (4-pyridylthio) -undecanoic acid (6) In the same manner as in Example 1, 11- (4-pyridylthio)-was prepared from 1 g of 11-undecanoic acid bromide and 0.42 g of 4-mercaptopyridine. 0.90 g of undecanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
1-1.51 (m, 14H, SCH ₂ CH ₂ ( CH ₂ )
₇ ), 1.63 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.17 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.05 (t, 2H, J = 7.
3 Hz, SCH ₂ ), 7.23 (d, 2H, J = 6.3)
Hz, pyridyl-H-3,5), 8.36 (d, 2H,
J = 6.3 Hz, Pyridyl-H-2,6), 11.98.
(Br s, 1H, COOH) UV: λmax (nm)
= 230,300

Example 7 Preparation of 9- (2-pyrimidylthio) -nonanoic acid (7) In the same manner as in Example 1, 9- (2-pyrimidylthio) was obtained from 1 g of 9-nonanoic acid bromide and 0.47 g of 2-mercaptopyrimidine.
-0.80 g of nonanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
1-1.52 (m, 10H, SCH ₂ CH ₂ ( CH ₂ )
₅ ), 1.63 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.17 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.10 (t, 2H, J = 7.
3Hz, SCH ₂ ), 7.18 (t, 1H, J = 5.0
Hz, pyrimidyl-H-5), 8.63 (d, 2H, J
= 5.0 Hz, pyrimidyl-H-4,6), 11.96
(Br s, 1H, COOH) UV: λmax (nm)
= 250

Example 8 Preparation of 10- (2-pyrimidylthio) -decanoic acid (8) In the same manner as in Example 1, 10- (2-pyrimidylthio)-was obtained from 1 g of 10-bromodecanoic acid and 0.45 g of 4-mercaptopyrimidine. 0.81 g of decanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
_{0-1.50 (m, 12H, SCH 2} CH 2 (CH 2)
₆ ), 1.62 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.15 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.06 (t, 2H, J = 7.
3Hz, SCH ₂ ), 7.16 (t, 1H, J = 5.0
Hz, pyrimidyl-H-5), 8.59 (d, 2H, J
= 5.0 Hz, pyrimidyl-H-4,6), 11.96
(Br s, 1H, COOH) UV: λmax (nm)
= 250

Example 9 Preparation of 11- (2-pyrimidylthio) -undecanoic acid (9) In the same manner as in Example 1, 11- (2-pyrimidylthio)-was prepared from 1 g of 11-undecanoic acid bromide and 0.42 g of 4-mercaptopyrimidine. 0.84 g of undecanoic acid was obtained. ¹ H-NMR (DMSO-d6 / TMS): δ = 1.2
4-1.53 (m, 14H, SCH ₂ CH ₂ ( CH ₂ )
₇ ), 1.63 (quint, 2H, J = 7.3 Hz,
SCH ₂ CH ₂ ), 2.18 (t, 2H, J = 7.3H
z, CH ₂ COOH), 3.12 (t, 2H, J = 7.
3Hz, SCH ₂ ), 7.21 (t, 1H, J = 5.0
Hz, pyrimidyl-H-5), 8.62 (d, 2H, J
= 5.0 Hz, pyrimidyl-H-4,6), 11.97
(Br s, 1H, COOH) UV: λmax (nm)
= 250

Example 10 10- (4-Pyridylthio) -decanoyl-CoA (1
0) Preparation and chemical analysis Method of Heuckeroth et al. (Proc. Natl. Acad. Sci. USA, 8
5, 8795 (1988)), compound 5 and LiC
oA was reacted in the presence of an acyl-CoA synthetase at 30 ° C. for 1 hour and 30 minutes. The reaction solution was injected into a reverse phase C18 HPLC column and eluted under the linear gradient condition of 6-40% acetonitrile / 0.1% trifluoroacetic acid aqueous solution. The product was evaluated by measuring the absorption maximum wavelength of the fatty acid analog at 300 nm. The peak ratio of the compound 10 to the unreacted peak of the compound 5 was 99 or more.

Example 11 Nα-10- (4-pyridylthio) -decanoylglycyl-L-asparaginyl-L-alanyl-L-alanyl-L-alanyl-L-alanyl-L-arginyl-L-
Production of Arginine (11) Compound 5, N-hydroxysuccinimide and N, N′-
Dicyclohexylcarbodiimide was reacted in acetonitrile at room temperature overnight. An aqueous solution of glycyl-L-asparaginyl-L-alanyl-L-alanyl-L-alanyl-L-alanyl-L-arginyl-L-arginine separately synthesized by the obtained acetonitrile solution of active ester and solid-phase peptide synthesis was added to triethylamine. Was reacted in the presence of. The reaction mixture was injected onto a reverse phase C18 HPLC column and eluted under the condition of a linear gradient of 6-40% acetonitrile / 0.1% trifluoroacetic acid aqueous solution. Each fraction was analyzed by reverse phase C18 HPLC and PICO-
Scrutinized by amino acid composition analysis by TAG method,
10- (4-pyridylthio) -decanoylglycyl-L
-Asparaginyl-L-alanyl-L-alanyl-L-
Alanyl-L-alanyl-L-arginyl-L-arginine (11) was obtained. Amino acid composition analysis value (6M hydrochloric acid,
110 ° C, 24 hours) Gly (1.12), Asp (0.90), Ala
(3.78), Arg (2.19)

Test Example 1 N-myristoyltransferase (NMT) activity measurement Method of Heuckeroth et al. (Proc. Natl.
Acad. Sci. USA, 85, 8795 (198
8)), the CoA ester of the compound of the present invention prepared in the same manner as in Example 10, glycyl-L-asparaginyl-L-alanyl-L-alanyl-L-alanyl-L as a substrate.
-Alanyl-L-arginyl-L-arginine, as an enzyme, HTLV-I persistently infected cells MT-2 were subjected to cell fractionation according to a centrifugal fractionation method, and adjusted to 100,000 × gPpt.
The enzyme activity of NMT was measured using the fractions. The reaction solution was injected into a reverse phase C18 HPLC column and eluted under the linear gradient condition of 6-40% acetonitrile / 0.1% trifluoroacetic acid aqueous solution. The evaluation of the product was carried out by using the compound 11 chemically synthesized in Example 11 as an index and measuring the absorption maximum wavelength of the fatty acid analog at 300 nm. Unreacted 10- (4-pyridylthio) -decanoyl-CoA
The peak ratio of the compound 11 to the peak of (10) was 9
It was 9 or more.

Test Example 2 Measurement of Anti-HIV Activity Using Primary Infected Cells Shoji et al.'S method (Biochem. Biophys. Re)
s. Commun. , 194, 610 (1993)), human T-cells (CEM, 2 × 1) in the logarithmic growth phase.
0 5 cells / ml, 10 ml) so that the final concentration of the compound of the present invention is 100 μM.
RPMI-1640 was added to infect HIV-I virus (0.002 MOI (number of infectious virus particles / number of cells in culture)). 37 ° C, 1 hour after infection,
Washed twice with 10 ml of 10% FCS RPMI-1640 and containing 100 μM compound of the invention 10% FCS R
10 ml of PMI-1640 was added. The number of cells was counted every 24 hours by the trypan blue staining method to determine the toxicity of the compound. After centrifugation at 260 xg for 5 minutes, the culture supernatant was filtered with a 0.45 µm filter to determine the 50% virus infectious titer (TCID50). The cells recovered by centrifugation were 10% FC containing 100 μM compound of the present invention.
The culture was continued by resuspending with 10 ml of S RPMI-1640.

No cytotoxicity was observed at the above compound concentrations in the assay by the Trypan Bull-staining method.

Table 1 shows the results of determining the virus production inhibition rate after 48 hours based on the virus infectivity without drug. Table 1 Antiviral activity of fatty acid analogs using primary infected cells ─────────────────────────── Compound No Virus production inhibition rate (%) ─────────────────────────── 4 90.0 5 98.7 ──────────────── As is clear from Table 1, the fatty acid analog of the present invention suppresses virus production in the primary infection system.

Test Example 3 Measurement of anti-HIV activity on HIV-I persistently producing cells (CEM / LAV-I) Method by Shoji et al. (Biochem.
Biophys. Res. Commun. , 194, 6
10 (1993)), HIV-in log phase
I continuous production strain (CEM / LAV-I, 5 × 10 5 cel
ls / ml, 10 ml) with 0.001% P.I. O. E.
(60) FCS-free RPM containing hydrogenated castor oil
After washing twice with I-1640 to remove virus particles adhering to the cell surface, 1.25 × 10 6 cells were
FCS-free RPMI containing 1 mM compound of the present invention
It was resuspended in 1 ml of -1640 and treated at 37 ° C for 30 minutes. 9m so that the final concentration of the compound is 100 μM
1 of RPMI-1640 was added. In the same manner as in Test Example 2, the cytotoxicity of the compound was determined every 24 hours by the Trypan Bull-staining method, and the anti-HIV activity was determined by the virus infectivity titer assay.

No cytotoxicity was observed at the above compound concentrations in the Trypan Bull-stain assay.

Table 2 shows the results of determining the virus production inhibition rate based on the virus infectivity without drug. Table 2 Antiviral activity of fatty acid analogs using HIV-I persistently producing cells ────────────────────────────── Compound No. Virus production inhibition rate (%) ────────────────────────────── 5 70 ───────────── ─────────────────── As is clear from Table 2, it is understood that the fatty acid analog of the present invention suppresses the growth of infectious cell virus.

Continuation of the front page (51) Int.Cl. ⁶ Identification code Office reference number FI technical display location C07D 239/34 239/38 249/12 512 257/04 GH

Claims (5)

[Claims] 1. A formula (I) in _{Z-Y- (CH 2) xCOOR} ( wherein, Z is a pyridyl group, a pyrimidyl group, a triazolyl group or tetrazolyl group, Y an oxygen atom or a sulfur atom, R represents hydrogen or An alkyl group having 1 to 8 carbon atoms and x represents 7 to 11), and a salt compound usable as a medicine. 2. A method for measuring N-myristoyl transferase activity using a fatty acid analog represented by formula (I) as a substrate. 3. A compound having antiviral activity represented by formula (I). 4. A compound having antiretroviral activity represented by formula (I). 5. A compound having anti-immune deficiency syndrome (AIDS) virus (HIV) activity represented by formula (I).