JPH08198826A - Compounds with antiviral activity - Google Patents

Abstract

(57) [Summary] [Object] The present invention provides a useful N-myristoyltransferase inhibitor for exploring the biological significance of myristoylation, and also provides a virus, a retrovirus, especially a human immunodeficiency syndrome ( Virus that causes AIDS (HIV-
Compounds having antiviral activity against I) are provided. The present invention provides a compound of formula (I) R _1- (CH ₂ ) x-CH ((CH ₂ ) y-NH-
R ₂₎ -CHO (wherein, R ₁ represents a hydrogen atom, a hydroxyl group, an alkyloxy group having 1 to 15 carbon atoms, represents a phenoxy group or benzyloxy group, R ₂ represents a hydrogen atom or an acyl group having 1 to 15 carbon atoms In which x represents 0 to 3 and y represents 0 to 3), an aminoalkyl aldehyde derivative or a sulfite adduct thereof, or a pharmaceutically usable salt compound thereof.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention has an N-myristoyltransferase inhibitory activity, and is a virus, a retrovirus, and particularly a causative virus (HI) of human immunodeficiency syndrome (AIDS).
The present invention relates to a novel aminoalkyl aldehyde derivative having excellent antiviral activity against VI) or a sulfite adduct thereof or a salt compound thereof which can be used as a medicine.

[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. The fact that myristic acid, a long-chain fatty acid having 14 carbon atoms, is covalently bonded to the amino terminus of the protein via an acid amide bond,
In 1982, in the course 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)). Since then, the amino-terminal myristoyl group of various proteins has been identified. Amino-terminal myristoylated proteins can be roughly classified into cell-derived proteins and virus-derived proteins. The former includes cAMP-dependent protein kinase and phosphoprotein phosphatase calcineurin.
B, reduced nicotinamide-adenine dinucleotide (NADH) -cytochrome b ₅ reductase, BC
Intracellular protein of ₃ H ₁ muscle cell line, pp60
The p36K substrate protein of ^v-src (src-gene gene product, tyrosine kinase), intracellular protein in macrophages, insulin receptor, vinculin and the like are known.

The latter amino terminus undergoes myristoylation
The virus constituent proteins that
Moloney murine leukemia virus Pr65^gagCoata
Protein, gag-onc protein of FeSV, Moro
Knee murine leukemia virus tyrosine protein
, Rous sarcoma virus (RSV) pp6
0^{v-sr c}, Adult T-cell leukemia (ATL) virus (H
P19 of TLV-I)^gagProtein, human immunodeficiency
P17 of causative virus (HIV-I) of symptoms group (AIDS)
^gagOf proteins, polyomavirus and SV40
Envelope protein VP2, Mason-Pfizer (Mas
on-Pfizer) simian virus core protein,
Pre-S1 core protein of hepatitis B virus (HBV)
Quality, poliovirus core protein VP4 is known
There is. In addition, human oncogene (src-gene) product p
p60^c-src, Guanosine triphosphate (GTP)
The myristoyl group is also present in the α-subunit of the binding protein.
Was found, and myristoyl was also found in immunoglobulin
The attachment of groups has been suggested. As mentioned above, various cells
Internal protein, virus constituent protein, or carcinogenesis
Myristoylation has been found in gene products.

Enzymes that catalyze the amino terminal myristoylation of proteins are described in yeast (DA Towler et al., J. B.
iol. Chem. , 263, 1784 (1988))
And animal tissue (CJ Glover et al., Bioch.
em. J. , 250, 485 (1988)) and partially purified, but its enzymatic chemical properties are still unclear. However, this enzyme is known to catalyze the transfer of myristoyl group between myristoyl coenzyme A (CoA) as a myristoyl donor and N-terminal glycyl peptide as an acceptor. As a relatively short synthetic N-terminal glycyl peptide which can be an acceptor of N-myristoyl transferase, for example, Gly-Asn-Ala-A
la-Ala-Ala-Arg-Arg and Gly-
Asn-Ala-Ala-Ser-Tyr-Arg-A
rg is US Pat. Nos. 4,740,588 and 4,7.
78,878.

On the other hand, various biological significances of myristoylation have been discussed. For example, in the case of pp60 ^v-src of RSV, inhibition of myristoylation loses the ability to cause cell transformation (D. Pellman et al., Natu.
Re, 314, 374 (1985)). Further, in the poliovirus core protein VP4, it has been reported that myristoylation plays an important role in order for the virus to retain the capsid structure or in the process of sending its own RNA to the host cell (M. Chow). Et al., Nature, 327, 482.
(1987)). S. P. Adams et al. Have oxy or thio fatty acids of 13 or 14 carbons in which the 4th to 13th carbons have been replaced by oxygen or sulfur (US Pat.
2,094) or an azido-substituted oxy or thiofatty acid having 9 to 13 carbon atoms and having an azide, tetrazoyl or triazolyl group at the ω position (US Pat. No. 59.
6,183) inhibit the enzymes that catalyze myristoylation and report that these fatty acid analogs inhibit the virus in virus-infected mammalian hosts.

[0006] In addition, Ikeda et al., In the fatty acid skeleton, pyridyl group, pyrimidyl group, triazolyl group, tetrazolyl group,
A fatty acid analog containing an oxygen atom or a sulfur atom (Japanese Patent Application No. 6-23541) has an inhibitory activity against an enzyme that catalyzes myristoylation.
It was reported that the virus production was inhibited when the strain was continuously produced by IV-I (CEM / LAV-I). All of the N-myristoyltransferase inhibitors reported to date are compounds having a carbon chain similar to myristic acid. As described above, the biological significance of myristoylation and the importance of antiviral activity against viruses and retroviruses due to N-myristoyl transferase inhibition, especially against the causative virus (HIV-I) of human immunodeficiency syndrome (AIDS) are clarified. Therefore, the development of new N-myristoyltransferase inhibitor compounds having stronger activity or different action mechanism is desired.

[0007]

SUMMARY OF THE INVENTION The present invention provides an excellent N-
It is an object of the present invention to provide a novel aminoalkyl aldehyde derivative having myristoyl transferase inhibitory activity. Further, the present invention aims to provide compounds having excellent antiviral activity.

[0008]

In order to achieve the above object, various aminoalkyl aldehyde derivatives, particularly aminopropanal derivatives and their sulfite adducts have been studied, and as a result, the formula (I) R _1- (CH ₂ ) x _{-CH ((CH 2) y-} NH-R 2) -CHO (I) ( wherein, R ₁ represents a hydrogen atom, a hydroxyl group, an alkyloxy group having 1 to 15 carbon atoms, a phenoxy group or benzyloxy group, R ₂ represents a hydrogen atom or an acyl group having 1 to 15 carbon atoms, x represents 0 to 3, and y represents 0 to 3), and an aminoalkyl aldehyde derivative represented by the formula is an excellent N-myristoyltransferase. It was found to exhibit inhibitory activity.

Further, the compounds of the present invention are CEM / LAV
It was confirmed to suppress the proliferation of HIV-I persistently infected cells in an assay using -I cells. This is because the compound of the present invention inhibits the action of N-myristoyl transferase, and therefore the cellular transformation, the retention of the viral capsid structure in the core protein, which has been discussed as the biological significance of myristoylation, or It is meant to change in the process of sending viral RNA to the host cell. That is, the compound of the present invention is useful as an N-myristoyl transferase inhibitor and an antiviral agent against viruses, retroviruses, and in particular, the causative virus (HIV-I) of human immunodeficiency syndrome (AIDS).

In the formula (I), preferred R ₁ is hydrogen atom, hydroxyl group, alkyloxy group having 1 to 4 carbon atoms or benzyloxy group, and R ₂ is hydrogen atom or 1 carbon atom.
-15 acyl groups, x is 1, and y is 0. More specifically, the following aminoalkyl aldehyde derivatives can be mentioned. 2-Aminopropanal sulfite adduct (1) 2-Amino-3-hydroxypropanal sulfite adduct (2) 2-Amino-3-tertiary butoxypropanal sulfite adduct (3) 2-Amino-3-benzyl Oxypropanal sulfite adduct (4) 2- (1-octanoylamino) -3-hydroxypropanal (5) 2- (1-dodecanoylamino) -3-hydroxypropanal (6) 2-myristoylamino- 3-Hydroxypropanal (7) 2-Myristoylamino-3-hydroxypropanal sulfite adduct (8) The present invention is not limited to these.

The compound of the present invention can be produced by various synthetic routes. For example, P. Garner et al. (J. Org. Chem., 52, 2361 (198).
In accordance with 7)), an aminoalkyl aldehyde derivative obtained by reducing an aminoalkylcarboxylic acid methyl ester derivative in which an amino group is protected with a suitable protecting group with diisobutylaluminum hydride (DIBAL) is added to a sulfurous acid gas or sulfurous acid gas in the presence of water. By acting
An aminoalkylaldehyde derivative free of an amino group can be obtained as a sulfite adduct. Further, an N-acylaminoalkyl aldehyde derivative can be obtained by similarly reducing the N-acylaminoalkylcarboxylic acid methyl ester derivative with a suitable reducing agent. The above reactions are examples, and other equivalent known methods can be used. The aminoalkyl aldehyde derivative or the sulfite adduct thereof of the present invention has one or two asymmetric carbon atoms, and any stereoisomers based thereon are included in the present invention.

Next, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto. Example 1 Preparation of 2-aminopropanal sulfite adduct (1) The method of Nahm et al. (Tetrahedron Le
tt. , 22, 3815 (1981)), N-tert-butoxycarbonyl-L-alanine (2.0 g) in 10 ml of methylene chloride was added to BOP reagent (4.68 g),
N, O-dimethylhydroxyamine hydrochloride (1.13
g) and triethylamine (1.8 ml) to give N-tert-butoxycarbonyl-L-alanine methoxymethylamide (1.89 g). The obtained compound (1.0 g) was added to J. A. According to the method of Fehrentz et al. (Synthesis, 676 (1983)),
It was dissolved in 20 ml of tetrahydrofuran, lithium aluminum hydride (0.2 g) was added under ice cooling, and the mixture was stirred for 1 hour. Aqueous potassium hydrogen sulfate solution (1.1 g /
(20 ml) was added to acidify and the mixture was extracted with ethyl acetate.
The extract was washed with 5% aqueous sodium hydrogen carbonate solution and then with saturated saline, and the ethyl acetate layer was dried over anhydrous magnesium sulfate, and the organic solvent was evaporated under reduced pressure to give 2-tertiary butoxycarbonylamino-3-pro- Panhard (0.41
g) was obtained.

The obtained aldehyde derivative (0.17 g)
Was suspended in 5 ml of distilled water at room temperature, a sulfurous acid gas was blown therein for 30 minutes, and then the mixture was stirred for 2 days under a sealed stopper. The reaction mixture was evaporated under reduced pressure, and a white solid precipitated by adding methanol was collected by filtration to obtain a 2-amino-3-propanal sulfite adduct (0.1 g). IR (KBr): 3126, 1367, 1347, 12
^{80,1186,1093,1061,1040cm -1 1 H-NMR (DMSO} -d6, δ): 1.24 (d
d, 3H, J = 3.63, 2.97 Hz, CH ₃ ),
3.30-3.54 (m, 1H, CH (α)), 3.9
7 (dd, 1H, J = 2.97 Hz, CH (β))),
4.57 (br, 1H, OH), 7.95 (br, 3
H, NH ₂ ) FABMS (m / z): 156 (M + 1) ⁺

Example 2 Preparation of 2-amino-3-hydroxypropanal sulfite adduct (2) P. Garner et al. (J. Org. Chem.,
52, 2361 (1987)).
Serine methyl ester hydrochloride (12.26 g) to 4-
Formyl-2,2-dimethyl-3-oxazolidinecarboxylic acid tert-butyl (5.6 g) was obtained. The obtained aldehyde derivative (3 g) was reacted with sulfurous acid gas in the same manner as in Example 1 to obtain a 2-amino-3-hydroxypropanal sulfite adduct (1.5 g). It was confirmed by FABMS measurement that the obtained compound formed a salt between two molecules. IR (KBr): 3448, 1701, 1367, 11
^{65,1037cm -1 1H-NMR (DMSO-} d6 +
D ₂ O / TMS): δ = 3.34 (m, 1H, CH
(Α)), 3.65 and 3.85 (each as
m, 2H, CH ₂ ), 4.21 (d, 1H, J = 6.3)
Hz, CH (β)) FABMS (m / z): 172 (M + 1) ⁺ , 343
(2M + 1) ⁺

Example 3 Production of 2-Amino-3-tert-butoxypropanal sulfite adduct (3) The reaction was carried out in the same manner as in Example 1, and N-tert-butoxycarbonyl-O-tert-butyl-L-serine ( 2-Amino-3-tertiary butoxypropanal sulfite addition by reacting 2-tertiary butoxycarbonylamino-3-tertiary butoxypropanal (1.0 g) obtained from 1.4 g) with sulfurous acid gas. The product (0.55 g) was obtained. IR (KBr): 3379, 1613, 1514, 13
^{65,1190,1040cm -1 1 H-NMR (DMSO} -d6 + D 2 O / TMS): δ
= 1.12 (s, 9H, tBu), 3.40-3.61
(M, 3H, CH (α), CH ₂ ), 4.16 (d, 1
H, J = 1.0 Hz, CH (β)) FABMS (m / z): 228 (M + 1) ⁺

Example 4 Preparation of 2-amino-3-benzyloxypropanal sulfite adduct (4) The reaction was carried out in the same manner as in Example 1 to give N-tert-butoxycarbonyl-O-benzyl-L-serine (1.4 g). 2-tert-butoxycarbonylamino-3-benzyloxypropanal (1.1 g) obtained from (2) and sulfurous acid gas were reacted to give 2-amino-3-benzyloxypropanal sulfite adduct (0.58 g). )was gotten. IR (KBr): 3171, 1586, 1507, 14
54, 1253, 1181, 1103, 1083, 10
^{26cm -1 1 H-NMR (DMSO} -d6 / TMS): δ = 3.5
1-3.77 (m, 3H, CH (α), CH ₂ ), 4.
26 (s, 1H, CH (β)), 4.62 (s, 2H,
_{PhCH 2), 7.37-7.47 (m,} 5H, A
r), 7.80 (br, 3H, NH ₂ ) FABMS (m / z): 262 (M + 1) ⁺

Example 5 Preparation of 2- (1-octanoylamino) -3-hydroxypropanal (5) The reaction was carried out in the same manner as in Example 1 to carry out N-tert-butoxycarbonyl-O-benzyl-L-serine (3). 0.8 g) gave N-tert-butoxycarbonyl-O-benzyl-L-serine methoxymethylamide (3.5 g). After treating the obtained compound with trifluoroacetic acid, it was reacted with octanoic acid chloride in the presence of triethylamine in chloroform to give N-1-octanoyl-O-benzyl-L-.
Serine methoxymethylamide was obtained quantitatively. N-1-Octanoyl-L-serine methoxymethylamide was quantitatively obtained by subjecting the obtained compound to catalytic reduction in methanol in the presence of 10% palladium-carbon. The obtained compound was dissolved in 10 ml of ether, lithium aluminum hydride (0.066 g) was added under ice cooling, and the mixture was stirred for 1 hour. The reaction mixture was acidified by adding an aqueous solution of potassium hydrogen sulfate (0.33 g / 10 ml), and the white solid precipitated in the ether layer was collected by filtration to obtain the desired 2-
(1-Octanoylamino) -3-hydroxypropanal (0.11 g) was obtained.

IR (KBr): 2956, 2854,
1643, 1547, 1469, 1421, 1385,
1332, 1247, 1146, 1097, 1078c
m ^-11 H-NMR (DMSO-d6 / TMS): δ = 0.8
7 (t, 3H, Me), 1.26 (s, 8H, C
_{H 2), 1.50 (br,} 2H, CH 2) 2.20-
_{2.22 (m, 2H, CH 2} ), 3.60-3.70
_{(M, 1H, CH 2)} , 3.77-3.85 (m, 1
H, CH ₂ ), 4.17 (dd, 1H, J = 4.16,
6.93 Hz, CH (α), 4.93 (t, 1H, O
H), 9.48 (s, 1H, CHO) FABMS (m / z): 217 (M + 1) ⁺

Example 6 Preparation of 2- (1-dodecanoylamino) -3-hydroxypropanal (6) The reaction was carried out in the same manner as in Example 5, and N-1-dodecanoyl-L-
Serine methoxymethylamide (0.8 g) to 2- (1
-Dodecanoylamino) -3-hydroxypropanal (0.083g) was obtained. IR (KBr): 2922, 2851, 1640, 16
17, 1547, 1471, 1249, 1147, 11
^{24,1085cm -1 1 H-NMR (DMSO} -d6 / TMS): δ = 0.8
6 (t, 3H, Me), 1.25 (s, 16H, C
H ₂ ), 1.48 (br, 2H, CH ₂ ), 2.05-
_{2.33 (m, 2H, CH 2} ), 3.57-3.81
(M, 2H, CH ₂ ), 4.16 (dd, 1H, J =
6.6 Hz, CH (α), 4.97 (br, 1H, N
H), 9.47 (s, 1H, CHO) FABMS (m / z): 272 (M + 1) ^+.

Example 7 Preparation of 2-myristoylamino-3-hydroxypropanal (7) Reaction was carried out in the same manner as in Example 5, and N-myristoyl-L-serine methoxymethylamide (0.6 g) to 2-myristoylamino- 3-hydroxypropanal (0.13
g) was obtained. IR (KBr): 3536, 3308, 2956, 29
20, 2851, 1640, 1618, 1546, 14
^{71,1147,1125cm -1 1 H-NMR (DMSO} -d6 / TMS): δ = 0.8
6 (t, 3H, Me), 1.24 (s, 20H, C
H ₂ ), 1.50 (br, 2H, CH ₂ ), 2.18
(T, 2H, CH ₂ ), 3.61-3.69 (m, 1
_{H, CH 2), 3.75-3.83 (} m, 1H, C
H ₂ ), 4.16 (dd, 1H, J = 6.27, 4.9)
5 Hz, CH (α), 4.96 (t, H, OH),
8.15 (d, 1H, J = 6.59Hz, NH), 9.
47 (s, 1H, CHO) FABMS (m / z): 300 (M + 1) ⁺

Example 8 Preparation of 2-myristoylamino-3-hydroxypropanal sulfite adduct (8) N-myristoyl-L-serine methoxymethylamide (0.6 g) obtained in the same manner as in Example 5 was dissolved in 8 ml of ether.
Lithium aluminum hydride (0.08g)
Was added under ice cooling and stirred for 1 hour. An aqueous potassium hydrogensulfate solution (0.4 g / 10 ml) was added to the reaction mixture to make it acidic, the white solid precipitated in the ether layer was filtered off, and then sulfurous acid gas was blown into the ether solution for 20 minutes. Petroleum ether was added to the reaction solution and the precipitated white solid was collected by filtration to obtain the desired 2-myristoylamino-3-hydroxypropanal sulfite adduct (0.16 g). IR (KBr): 2957, 2920, 2851, 16
18, 1546, 1471, 1214, 1147, 11
^{23,1109,1083,1037cm -1 1 H-NMR (DMSO} -d6 / TMS): δ = 0.8
6 (t, 3H, Me), 1.24 (s, 20H, C
H ₂ ), 1.46 (br, 2H, CH ₂ ), 2.03-
_{2.09 (m, 2H, CH 2} ), 3.33-3.82
_{(M, 6H, CH (2} ), CH 2, OH (2)) FABMS (m / z): 380 (M-1) -

Next, the N-myristoyl transferase (NMT) inhibitory activity test of the compound of the present invention and the anti-HIV activity measurement test for HIV-I persistently producing cells (CEM / LAV-I) are shown. Test Example 1 Measurement of N-myristoyl transferase (NMT) inhibitory activity of the compound of the present invention. O. Heuckeroth et al. (Proc. N
atl. Acad. Sci. USA, 85, 8795
(1988)), a myristoyl CoA ester prepared by reacting myristic acid and LiCoA in the presence of an acyl-CoA synthase at 30 ° C. for 1 hour and 30 minutes, and glycyl-L-asparaginyl-L-alanyl-L- as a substrate.
Alanyl-L-alanyl-L-alanyl-L-arginyl-L-arginine and HTLV-I persistently infected cells MT-2 as an enzyme were fractionated according to a centrifugal fractionation method to prepare 100,000 × gPpt fraction. Then, the enzyme activity of NMT was measured in the presence and absence of 20 μM of the compound of the present invention. The reaction solution was injected into 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, and N-myristoylglycyl-L-asparaginyl-of the enzyme reaction product was eluted. L-alanyl-L-alanyl-L-alanyl-
L-alanyl-L-arginyl-L-arginine was quantified. The results are shown in Table 1.

[0023]

[Table 1] As is clear from Table 1, the compound of the present invention inhibits N-myristoyl transferase (NMT) activity.

Test Example 2 Measurement of anti-HIV activity on HIV-I persistently producing cells (CEM / LAV-I) Method by Shoji et al. (Biochem. Biophys. Re)
s. Commun. , 194, 610 (1993)), the HIV-I continuous production strain (CEM) in the logarithmic growth phase.
/ LAV-1, 2 × 10 ⁵ cells / 10 ml), the final concentration of the compound of the present invention is 0, 60, 80, 100, 20.
The cells were added to 0 μM and cultured, and 72 hours later, the number of viable cells was measured by the trypan blue staining method. Table 2 shows the results. No cytotoxicity against normal cells (CEM) was observed at the above compound concentrations.

[0025]

[Table 2] As is clear from Table 2, the compounds of the present invention inhibit the growth of HIV-I persistently producing cells (CEM / LAV-I).

[0026]

According to the present invention, it has an inhibitory activity against an enzyme which catalyzes the amino-terminal myristoylation of protein,
It was possible to provide a novel aminoalkylaldehyde derivative which inhibits the growth of virus-infected cells and exhibits antiviral activity.

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI technical display location C07C 301/00 7419-4H 311/04 7419-4H

Claims (3)

[Claims] 1. Formula (I) R _1- (CH ₂ ) x-CH ((CH ₂ ) y-NH-R ₂ ) -CHO (I) (In the formula, R ₁ is a hydrogen atom, a hydroxyl group, or the number of carbon atoms. 1 to 15 represents an alkyloxy group, a phenoxy group or a benzyloxy group, R ₂ represents a hydrogen atom or an acyl group having 1 to 15 carbon atoms, x represents 0 to 3, and y represents 0 to 3 ) Or a sulfite adduct thereof or a salt compound thereof usable as a medicine. 2. Formula (II) R ₁ —CH ₂ CH (NH—R ₂ ) —CHO (II) (In the formula, R ₁ is a hydrogen atom, a hydroxyl group, an alkyloxy group having 1 to 4 carbon atoms or benzyloxy. Group represented by the formula, R ₂ represents a hydrogen atom or an acyl group having 1 to 15 carbon atoms), a sulfite adduct thereof or a pharmaceutically usable salt compound thereof. 3. An N-myristoyl transferase inhibitor containing the compound according to claim 1 or 2 as an active ingredient.