JPH013185A - Active compounds, their preparation and pharmaceutical compositions containing them - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates to an isomeric form of a chemical compound having antiviral activity, to a process for the preparation of this isomer, and to their use as a medicine.

[Conventional technology]

European Patent Publication No. 242,482 (the main points of which are incorporated herein by reference) describes the formula (A) in which R+ is hydrogen or CH20H; R+ is hydrogen or (When R1 is hydrogen) Hydroxyl or CHz OH υ; R3 is CH20H or (When R and R2 are both hydrogen) CH(OH) CH2-OH"'C'
;h;R4 is hydrogen, hydroxyl, amino or FiO&
<In the formula, R6a' Ct~6 alkyl, phenyl or haphenyl C1-2 alkyl, and any of the phenyl moieties is 1 or 2 halogens, C1-4 alkyl or C1
and R1 # any OH group of R2 and/or R3 may be in the form of its O-acetyl, phosphate, cyclic acetal or cyclic carbonate conductor ] and pharmaceutically acceptable salts thereof are disclosed.

Example 7 describes the (R,S) racemic mixture of the compound 2-amino-9-(2,3-dihydroxyprob-1-oxy)purine, and Example 30 is prepared by stereospecific synthesis. The pure (R)-isomer obtained is described. The pure (S)-isomer has now been prepared and is found to have greater activity as an antiviral agent compared to the (R)-isomer. It is believed to be a prodrug of a compound of formula (A) that is OH. European Patent Publication No. 2424
Example 31 of Specification No. 82 (S) -9-(2,3-
dihydroxyprob-1-oxy)guanine is described.

[Summary of the invention]

Therefore, the present invention can handle up to a large amount of 45N in the mixture (R)
- (S ) - provides isomers.

0-Acyl derivatives are usually optionally substituted at one or both of the OH groups in the side chain by a carboxylic acid ester group, such as one or two C1-4 alkyl, C1-4 alkoxy, halogen or CFx groups. It forms good CI~7 alkanoyl and benzoyl. Preferably the carboxylic acid ester group is a C1-7 alkanoyl group such as acetyl, propionyl, butyryl, heptanoyl and hexanoyl, most preferably acetyl or propionyl.

Examples of phosphate esters of compounds of formula (1) are those in which one of the acyclic -OH groups is replaced by a (HO)z-PO2- group or a salt thereof or two 10H groups are bridged 10 -P (OH) Including those substituted with 02- group.

The two acyclic OH groups can also be substituted by radicals such as cyclic acetal groups such as -0-C(C+~3alkyl)2-0- or cyclic carbonates such as -o-co-o-.

Preferably the (S)-isomer of the compound of formula (1) as defined or a derivative thereof is 0 to 40 of the corresponding (R)-isomer.
The form includes 0 to 30 inches, 0 to 20 inches, or 0 to 10 inches. More preferably, the (S)-isomer is in a form containing 0 to 5% of the corresponding (R)-isomer.

Most preferred are forms containing zero (S)-isomer or undetectable amounts of the corresponding (R)-isomer. All values above are in percent by weight of the mixture. The presence of the (R)-isomer can be determined, for example, by comparing the optical rotation of a sample of the isomer mixture with that of a pure sample of the (S)-isomer or of a sample of the isomer mixture in the presence of a xyl shift reagent or xyl solvating agent. It can be conventionally detected by IHnmr.

The present invention provides a method for preparing the (S)-isomer or derivative thereof of a compound of formula (1) as defined, the method comprising:
II) The compound of IOR(n) (wherein R and Rb are hydrogen or a protecting group) may be reduced and then optionally converted to R and/or RbeOH or as defined in formula (1). The compound may form derivatives thereof and/or may form pharmaceutically acceptable salts thereof.

Reduction may preferably be achieved using catalytic methods such as palladium or charcoal in an inert solvent such as methanol or ethanol at reflux temperature. The hydrogen source may be cyclohexene or more preferably ammonium formate.

One suitable protecting group is when R and Rb together are 1,3
- forms a dioquinrane ring (produced by reacting a corresponding compound in which R and Rb are H with 2,2-dimethoxypropane). This group can be removed by acid hydrolysis if desired. It will be understood that compounds of formula (1) in which R and Rb form a 1.3-dioxolane ring are active compounds within the scope of Wenmu (]).

Pharmaceutically acceptable salts, O-acyl derivatives and phosphate derivatives are conventionally prepared and are described, for example, in European Patent Publication Nos. 141927 and 182024.

Acyl derivatives of compounds of formula (1) can be prepared by deprotecting the products obtained, as is well known to those skilled in the art.

The acylation reaction may be carried out using an acylating agent containing a suitable carboxylic acyl group.

Examples of acylating agents suitable for the aforementioned process are carboxylic acids, acid halides such as chlorides or acid anhydrides, preferably anhydrides or acids.

When the acylating agent is a carboxylic acid, dicyclohexylcarbodiimide is included as a condensation accelerator, but this is unnecessary when the acylating agent is an acid anhydride.

Acylation reactions can be performed using a single acyl derivative or a mixture of derivatives of the compound of formula (1), depending on a number of factors, such as the relative amounts and chemical nature of the reactants, the physical conditions of the reaction and the solvent system. It can be generated.

Any mixture produced in this manner can be separated into its pure components using standard chromatography techniques.

The above-described acylation methods of the invention may produce mono- or di-acylated derivatives of the components of formula (1) depending on the type of protection/deprotection used. Examples of products obtained by different methods are shown below.

(a) Acylated derivatives of the OH group can be obtained by direct acylation of compounds of formula (1) when both acyl groups are identical.

(b) Mono-acylated derivatives of one OH group are used in the acylation of protected intermediates of compounds of formula (1) in which the other -OH group is preferably protected, for example by a monomethoxytrityl or trityl group. It can then be obtained by subsequent deprotection by acid treatment. Diacylated derivatives with different acyl groups can then be prepared as in (a) K.

Acyl derivatives of compounds of formula (1) may be converted to compounds of formula (1) by conventional deacylation or partial deacylation methods. For example, reaction with methanol and ammonia can be used to effect complete deacylation to yield compounds of formula (1) in which both OH groups are deacylated. Reaction with a mild base such as potassium carbonate results in partial deacylation of the diacylated derivative, leaving one acyl group and one
Compounds of formula (1) can be produced in which 2 OH groups are present.

Phosphate derivatives are formed by reaction with phosphorylating agents such as phosphorus oxychloride in vizidine. The NHK and OH groups are optionally or optionally protected using trityl or methoxytrityl protecting groups which are preferably removable by acid hydrolysis using acetic acid.

A cyclic phosphate derivative is generated from phosphorus oxychloride when both OH groups are phosphorylated.

Another suitable phosphorylating agent is cyanoethyl phosphate, in which case the product is usually treated with aqueous ammonia, which produces the ammonium salt of the phosphate ester as the final product.

Monophosphates can be converted to cyclic phosphates with a dehydrating agent such as dicyclohexylcarbodiimide.

Cyclic acetal derivatives of the compound of formula (1) are acyclic acetals such as Rlo O-C(C1-3 alkyl)2-
OR1o (wherein RIO is 01-4 alkyl such as methyl or ethyl) can be prepared from compounds of formula (1). The reaction is preferably carried out in the presence of a silicic acid such as p-)luenesulfonic acid in an inert solvent such as tetrahydrofuran or dimethylformamide.

The cyclic carbonate derivative of the compound of formula (1) is converted to formula (1) by phosgene or 1,1-carbonyldiimidazole.
(wherein the -NHK group is preferably protected) and then deprotected if necessary. Suitable protecting groups include trityl and monomethoxytrityl. The reaction is preferably carried out in dry pyridine, preferably at ambient temperature from 0 to 50<0>C.

It will be appreciated that deprotection and formation of derivatives may occur in any desired or necessary order.

Compounds of formula (II) may be prepared as follows.

Compounds of formula (■) are either well known or prepared by methods similar to those used for standard structurally similar well known compounds. Compounds of formula (V) can be prepared using formic acid and acetic anhydride.

(Org, Chem, ) j 40 (21),
2, which is a compound described in 3141.1975.
It can be prepared by formylation of 5-diamino-4,6-dichloropyridine.

The compounds of formula (II), (1), (IV) and (■) are new and form an aspect of the invention.

The present invention also provides a process for preparing the (S)-isomer of a compound of formula (1), optionally in combination with the corresponding (R)-isomer at 45% by weight S-t of the mixture; The method comprises preparing a mixture of the (S)- and (R)-isomers of the compound of formula (1), preferably a separated fraction rich in the (S)-isomer, and a separated fraction containing the corresponding (R)-isomer. 4
and may form arbitrarily defined derivatives and/or pharmaceutically acceptable salts thereof.

The term "resolution" is used herein in its conventional practical sense in the art, i.e., the separation of a mixture of enantiomers (in any ratio) of a compound into two fractions, one of which is initially contains a resolution that is enriched in one enantiomer compared to a mixture of

The resolution of a mixture of (S)- and (R)-isomers of a compound of formula (1) is achieved by derivatizing the mixture with a chiral derivatizing agent to separate the diastereoisomers of a derivative of a compound of formula (1). This can be done conventionally by forming a mixture. The components of the mixture can then be conventionally separated, for example by fractional crystallization. The separation may be complete or partial, the separated fraction consisting of diastereoisomeric derivatives of the (R)-isomer accounting for up to 45 q6 by weight of the total fraction. The desired diastereoisomeric derivative of the (S)-isomer, optionally mixed with the diastereoisomeric derivative of the (R)-isomer, is then optionally mixed with the (R)-isomer up to 45 times. The desired (S)- of the compound of formula (1) which may be mixed with
converted into isomers.

Derivatization in connection with the process of the invention preferably comprises (S )- and (R)-isomer mixtures. Such reagents include optically active acids or acid derivatives such as acid chlorides, acid anhydrides or isocyanates.

Salts and derivatives can be prepared as described above.

Racemic mixtures of isomers can be prepared by preparing the racemic intermediate of formula (■) according to the above or European Patent Publication No. 2424.
It can be manufactured according to the method described in Example 7 of the '82 specification.

A preferred method of preparing the pure (S)-isomer of formula (1), however, starts from a xyl intermediate of formula (II).

The compounds of the invention are useful in the treatment of infections caused by viruses, particularly herpesviruses such as herpes simplex type 1, herpes simplex type 2; Vallicella seastar virus; and also lentiviruses such as Visna virus.

Compounds of the invention may be formulated for use in pharmaceutical compositions. Accordingly, in another aspect of the invention there is provided a pharmaceutical composition comprising a compound of the invention together with a pharmaceutically acceptable carrier or excipient.

Compositions that can be administered to humans by the oral route can be in the form of syrups, tablets or capsules. When the composition is in tablet form, any pharmaceutical carrier suitable for formulating such solid compositions may be used, examples of which include magnesium stearate, starch, lactose, glucose, rice flour, wheat flour and chalk. . The composition may also be in the form of a digestible capsule containing the compound, eg made of gelatin, or a syrup, solution or suspension. Suitable liquid pharmaceutical carriers include ethyl alcohol, glycerin, saline and water, to which flavoring or coloring agents may be added to form a syrup. The compounds may also be provided with a sterile liquid carrier for injection.

The compositions may also be formulated for topical application to the skin or eyes.

For topical application to the skin, the composition may be in the form of a cream, lotion or ointment. These formulations can be found, for example, in standard books on pharmaceuticals and cosmetics, such as Leonard Hill Books.
"Harry's Cosmeticology" published by
ry's Cosmeticology) J and the British Pharmacopoeia, and may be conventional formulations well known to those skilled in the art.

Compositions for application to the eye can be conventional eye drop or ointment compositions well known to those skilled in the art.

Preferably, the compositions of the invention are in unit dosage form or other form that can be administered in a single dose.

A suitable dosage unit will contain from 9 to 1 g of 5Q m, e.g. 100 to 500 g, of active ingredient.

Such dosages may be administered one to four times per day or more usually two or three times per day. Effective dosages of the compounds will generally be within the range of 1.0 to 20 m9/kg body weight per day, or more usually 2.0 to 10 m/kg per day.

No positive effects are shown at the above-mentioned dosage levels.

The invention also provides a method of treating a viral infection in a human or non-human animal, which comprises administering to the animal an effective and non-toxic amount of a compound of the invention.

The invention also provides compounds of the invention for use as active therapeutic agents, particularly for the treatment of viral infections.

The compounds of the present invention also exhibit synergistic antiviral action in combination with interferons; and combination products containing these two components for sequential or intermittent administration by the same or different routes are a number of the invention. is within the range of

〔Example〕

The examples below illustrate the invention and the reference examples below illustrate the preparation of intermediates.

(R)-2,2 in tetrahydrofuran (200m)
-dimethyl-1,3-dioxolane-4-methanol (
7,39,0,05 mol), N-hydroxyphthalimide (8,15,!i', 0.05 mol) and triphenylphosphine (13,11,!9, 0.05 mol) t-0 C. and stirred during addition of diethyl azodicarboxylate (9.57.9°, 0.066 mol). The deep red solution was stirred for a total of 18 hours at 25°C until the color changed to pale yellow and the solution was evaporated to dryness. The residue was extracted once with ether (200 IILt) and the ether was removed under reduced pressure. The residue was dissolved in chloroform/hexane (1
0:1) to give (S)-N-(2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)phthalimide (10,5 g) as a white solid.
.

69ch)'t-obtained.

m, p. 99-100℃# ('') p'' -14,
6' (0,4gMeOH)yνmax (KBr
) (α)D” 1790.1730 and 1619cm
−”; δH(CD(Js) 1.35 (3H,5-C
Hs), 1.41(3H-8-CHs)3.99r
IH=(1-J5.5, 8.8Hz-CHzO
N/ChzOCMex) #4.17 (2H+m5
CH mushroom OCMe goose CHs ON + IH)
, 4.32 (IH, q-J 5.7, 10.1Hz
, CHsON/CH20Giiez) #4
．． 50 (IH, m, CH) and 7.75-7.87 (4
H, m, aromatic).

Actual measurements: C, 60,53; H, 5,46; N, 5.
03%, C14HC14H1 as C,60,64;H
,5,45;N,5.05chi.

(b) (S) -2,2-dimethyl-1,3-dioxolane dichloromethane (150IIlt)
-N-(2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)phthalimide (10 g, 36 mmol) and N-methylhydrazine (3,32,F, 7
2 mmol) was stirred at 5°C for t-2 hours. The reaction was filtered and evaporated to dryness and the residue was suspended in ether.

The suspension was filtered and evaporated and the residue was chromatographed in ethyl acetate as a clear liquid (S)-2,2-
Dimethyl-1,3-dioxolan-4-ylmethoxyamine (4,8,!i', 91%) was obtained.

(α)D"-2,4@(0,49sMeOH); νm
ax (film) 3550.3300 and 1600 equipment-1; δH (CDCls) 1.38 (3H, S.

CHs), 1.44 (3H, S BaJs), 3.
71 (3H, m, CHzOCMez CHzON+IH
)-4,05(IH-(1-J6.3,8.2Hz,C
IH of HzOCMez), 4.35 (IH,m,
CE()′IMj5.57 (2H,br.s,
DzO non-exchangeable, NHt); m/z132 (M”-CH
5e24%).

Actual value C, 48,76; H, 9, OO; N, 9.56%
C6H1sNO3 as C,48,96; H,8,90
;N, 9.52% (c) (S)-4-chloro-2
4,6-dichloro-2,5-diformamidopyrimidine (7,3&, 31.1 mmol) in ,5-diformamide diglyme (125 m), (S)-2,2
- A mixture of dimethyl-1,3-dioxolan-4-ylmethoxyamine (4,6&, 31.1 mmol) and diimpropylethylamine (8,03, iit, 62,1 mmol) was stirred at 100 °C for 4 hours. . The reactant is F
evaporated and the residue was chromatographed twice in chloroform-methanol (15:1) (S)-
4-chloro-2,5-diformamide-6-(2,2-
Dimethyl-1,3-dioxolan-4-ylmethoxyamino)pyrimidine C2.6g, 24th) was obtained 0 νmax(CHCls) 3400.1700.1580
．． 1560 and 1470crtr-"; δHCCDC
Jls) 1.39 (3H, S, CHs), 1.48 (
3H.

S-CHs) e 3.60-4.65 (5H#
m -CH atmosphereONH+CT(t0C2des 1CHOCMaz) e 7.96 (I Ha b
r, m e Ih O exchangeable, NH).

8.34 (IH, S, CHO), 9.25 (2H, br
, m, DgO exchangeable.

2XNH) and 9.50 (I H-S-CHO)
; m/'Z 344 (M"-H-<1ch), 33
0(M-CH5*<1ch).

Jetkimetchi, acetate (3α) +7) (S)
-4-chloro-2,5-diformamide-6-(2,2
-dimethyl-1,3-dioxolan-4-ylmethoxyamino)pyrimidine (2,1,!ii', 6.08
A solution of 0.0 mmol) was stirred at 120° C. for 3 hours. The reactants were evaporated to dryness, leaving a residue of methanol (40/d) and 0.8
8 dissolved in ammonia (IIlt) and 1.5 at 5°C.
Stir for hours. Evaporate and ethyl acetate-hexane (
The residue was chromatographed on silica eluting with 1:1) to give (S)-6-chloro-9 as a white solid.
-(2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)-2-formamide purine (1,3 g, 6
5 h) t-obtained.

m, p, 147-8℃; Shima:x (IG3r)
3420, 3200, 1700.

1610, 1580, 151ONj1440tan-1
; δH (CD (J!s) 1.40 (3H, S, CHs)
, 1.44 (3H=S-CHs), 3.89 (IH, m
CH! 0CIVIe! ) e 4.16 (I
Hs m s CH20CMez ) t 4.50
(3Hjm +CH+CH20N), 8.23(1,H
, S, H-8), 8.45 (IHebr, d.

Jll, 5Hz, DpO exchangeable, NH) and 9.59
(IH, d, Jll, 5Hz, HCONH).

Actual value C, 43,75; H, 4,29; N, 21.23
Person; M”327.0734.; Cxz HI3 ClN
504 as C, 43,98; H, 4,31; N121
．． 37%; M+327.0734° methanol (10 Inl) and 0.88 ammonia (1
(S) -6-ri0 low 9- (2,
2-dimethyl-1,3-dioxolan-4-ylmethoxy)-2-formamide purine (400m9.1.22
mmol) solution was stirred for 4 hours at 5°C. The solution was evaporated under reduced pressure and the residue was dissolved in ethyl acetate/hexane (2:1
)-2-amino-6-chloro-9- as a white solid by chromatography on silica that dissolves
(2,2-dimethyl-1,3-dioxolan-4-ylmethoxy)purine (270 mg, 74%) was obtained.

m, p, 118-120℃, νmax(KBr)34
50t3320°3210.1650,1630.16
20,1560.1510 and 1470cm-"; δ
H (CDCls) 1.36 (3H, S, CHs),
1.42 (3H.

S e CT(s) $ 3.85 (I Hlrn
t CHx OCMez ) t 4.14 (1,
H* m +CHsOCMez), 4.43 (
3H,m, CH+ CHzON), 5.51
(2H.

br, 5eDxo exchangeable, bait (!), 8.00
(IH, S, H-8).

Actual measurements: C, 44,45; H, 4,69; N, 23.
31%; M+299.0795; CuH14CINs
Os as C, 44,08; H, 4,71; N, 23.
37%; M+299.0785°80 (S)-2-amino-6 in thiacetic acid (201d)
-Chloro-9-(2,2-dimethyl-1,3-dioquinran-4-ylmethoxy)purine (250m9.0.8
A solution of 3 mmol) was stirred for 2 hours at 5°C and then for 1 hour at 70°C.
Stir at ℃. The solution was evaporated to dryness under reduced pressure, the residue absorbed on silica and chromatographed as a white solid, eluting with acetone-hexane (3:1) (S)-
2-amino-6-chloro-9-(2,3-dihydroxyprobo-1-oxy)purine (170 μm, 78%) was obtained.

m, p, 155-8°C, I'max (KBr) 334
0.3200.1660°1620, 1560, 1
520 traces 1470σ-1; δH ((CDs) Tomi SO)
3.41 (2H, m, CH20H) 3.78 (
IH, m, CH), 4.19 (IH-(IJ7
．． 4, 10.4Hz, CHZON) -4,4
0(IH-(1-J3.3, 10.4Hz.

CHzON), 4.73 (1'H,t, J5
．． 0Hz, D! 0 exchangeable, OH).

5.14 (IH, d, J5.OHz, D*O exchangeable,
OH), 7.12 (2H.

S, D*O exchangeable, NI (z) 8.35 (LH, S,
H-8).

Actual value; M+259.0475; C5H1oCA
! M+259.0472° as N5Os Example OH (S)-2-amino-6-chloro-9-(2,3-dihydroxyprofol-1-oxy)purine (160 μ, 0.62 mmol) in methanol (5a) ), ammonium formate (156μ, 2.5mmol) and arsenic charcoal (20+n9
) Heat the mixture of 10% palladium on t-3 to reflux for 3 hours. Next, ammonium formate (150 mfi'
, 2.5 mmol) and 10 on charcoal (20Tn9)
Chipalladium was added and the reaction was again heated to reflux for 1 hour. The mixture was filtered hot through glass fibers.
The liquid was evaporated under reduced pressure. The residue was recrystallized from hot ethanol to give (S)-2-amino-9-(2,
3-dihydroxyprobo-1-oxy)purine (100
■, 72%) was obtained.

m, p, 153-5°G, (α)D25+16.5°
(0,2, HzO);λmax(BhO)221(1
25,000), 305 (17250) nm Niji max
(KBr)3370-3310-3190=1650s
1620-1580゜1520.1480 and 1440
an-”; δH((CDs)tso) 3.43
(2Hzms CHzOH), 3.77 (IH,
m, CH), 4.18 (IH, (1-J 7.
8s 9.7Hz -CHZON) -4,40(
IH-q-J2.5-10.7Hz -CE'IxON
), 4.72 (IH, t, J5.4Hz, DtO exchangeable, OH).

5.16 (IH, d, J4.9Hz, Dzo replaceable,
OH), 6.72(2'f(.

S=D! 0 exchangeable, NH2), 8.27 (IH, 5-
H-8)-8,59 (IH, S, H-6).

Actual measurements: C, 41,26; H, 4,93; N, 29.
50 CH; C5HnNsOs, C as 0.5 HzO
,41,02;H,5,16;N,29.89chi.

Sample Example OH (S)-6-chloro-9-(2,2-dimethyl-1,3-dioquinrane- in 80% aqueous formic acid solution (121 m)
4-ylmethoxy)-2-formamide purine (300
7n'? , 0.92 mmol) was stirred at 100°C for 22.5 hours. The solvent was removed under reduced pressure, the residue was dissolved in methanol (2! nl) and 0.88 ammonia (2 resistant) and the solution was stirred for 2 hours at 5°C. The reaction was evaporated to dryness and the residue was dissolved in a minimum amount of hot water and filtered through a glass fiber vaporizer. The p-liquid was crystallized and the deposited solid was recrystallized in water to form a pale yellow solid (S)-9-(2,3-dihydroxyprobol 1).
-oxy)guanine (100■.

45%).

m, p, 252-5℃ (decomposition) [α] Du+13゜5℃
(0,16°Hg0); Jmax(HtO)252(
112,900) nm; umax (KBr) 3330.
3180, 1690, 1640.1600 and 1540
an-"; δ) (((CDs hso) 3.39
(2Hsms CH*01N), 3.73 (IH
,m.

CHOH), 4.10 (IH-(1-J7.7
, 10.5Hz, CHg0N), 4.32(
IH = (1-J 3.3, 10.5Hz, CH
20N), 4.70(IH,t, J5.6Hz
-D! O exchangeable, OH), 5.15 (IH, d, J
5. OHz, DgO exchangeable eOH) C6, 62 (2H
*br, 8eDso replaceable, Nflt).

7.90 (IH, 5=H-8), 10.64 (IH, b
r, S, and DiO can be exchanged.

NH); m/z 241 (M+, 13%).

Actual measurement: C, 38, 89; H, 4, 81; N, 28
．． 09%; M+241.0820; C5HnNsO
a, 0.4 HzO as C,38,68;H,4
, 79; N, 28.19; M 241.0811° This compound appears to be the active metabolite of the example compound.

Cells (Vero or MRC-5) were grown in aggregates in well-coated multi-dishes (hole diameter = 1.5 t:m). 100 μl of each drained cell monolayer
Herpes simplex virus 1 (H8V-1; H
FEM strain) or herpes simplex virus 2 (H3V −
2; MS strain). Virus was allowed to adsorb for 1 hour at room temperature. After adsorption, the remaining inoculum was removed from each well and added with 5 ml of newborn calf serum and 0.9% agarose (
It was replaced with Eagle's MEMO, 5-resistant containing A37). Once the agarose has solidified, Eagle's MIli
A dilution of the test compound prepared in M (containing 5% newborn calf serum) was added and each well was overlaid with 0.5-mL of liquid. Test compounds were diluted into the following concentration series. 200, 60, 20°6...0.06
μg/mt o Therefore the concentration in the assay is 100μ,!
It ranged between 9/d and 0.03 μg2. Infected cultures were incubated at 37°C in a humidified atmosphere at 5% C (h) until plaques were clearly visible (2 or 3 days for Vero cells, usually 1 day for MfLC-5 cells).

Cultures were fixed in formal saline, the agarose overlay was carefully washed away, and the cell monolayers were stained with carboluckin. Plaques were counted using a stereomicroscope. The ICBO (concentration of drug that inhibits the number of plaques by 50% relative to the number of plaques observed in the virus control monolayer) of the test compound was calculated. Furthermore, the monolayers were examined for drug-induced cytotoxicity and the lowest concentration at which cytotoxicity occurred was recorded.

2. CPE inhibition test against lentivirus (established monolayer) 3 x 10' sheep choroidal gland (SCP) cells were heat-activated with fetal calf serum (Fe2)'
Eagle's MEM1 with Hank's Salt containing tlo%
00 m into individual wells of a 96-well microtiter plate. When a monolayer becomes established (growth 1
or 2 days later) in 2004 storage medium (0,5%
Eagle's M with Hank's Salt containing Fe2'lr
EM) and “C” storage medium (30TCIDso
/ Season) was infected with the middle throat virus (K184 strain) ioon. The test sample was diluted in 3-fold dilution steps to other 9
Diluted in storage medium in a 6-well microtiter plate.

The diluted sample of 1004 was then transferred directly to the virus-infected monolayer, so the final concentration was 100-0.
, 0454/rlLt K), and socitevirus-induced CPE compared to untreated virus-infected controls (usually 1
The sample was incubated at 37° C. in a humidified 5-inch CO2 incubation until maximum (2-14 days). Plates were fixed with formal saline and stained with crystal violet). Virus-induced CPE was then counted microscopically and the minimum concentration (MIC) of the sample that completely protected the cell monolayer was determined.

The results regarding (S)-9-(2,3-dihydroxyprobo-1-oxy)guanine, which is the compound of Sample Example, were as follows.

IC KanT (SV-1 (5C16 strain, MRC-5 cells) 0.7
Tama EV-2 (MS strain, ■-5 cells) 0.06 Visna virus (Ni-184 strain in SCP cells) 10 This compound had no cytotoxicity on uninfected cells at concentrations up to 10,047 ml.

Agent Patent Attorney Masaaki Aki Sawa 1 other person

Claims (10)

[Claims] (1) Formula (I) which may be optionally mixed with the corresponding (R)-isomer up to 45% by weight of the mixture ▲Mathematical formulas, chemical formulas, tables, etc.▼(I) The (S)-isomer of the compound (the OH group may optionally be in the form of its O-acyl, phosphate, cyclic acetal or cyclic carbonate derivative) (2) A compound according to claim 1, wherein the compound of formula (I) is optionally in the form of a C_1-_7 alkanoyl ester derivative. (3) (S)-isomer of the compound of formula (I) or a derivative thereof according to claim 1 or 2, in a form containing 0 to 40% of the corresponding (R)-isomer. (4) The (S)-isomer of the compound of formula (I) or a derivative thereof according to claim 1 or 2, in a form containing 0 to 5% of the corresponding (R)-isomer. (5) The (S)-isomer of claim 4 in a form containing 0% or an undetectable amount of the corresponding (R)-isomer. (6) Formula (II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼(II) Reduce the compound (in the formula, R_a and R_b are hydrogen or a protecting group), and then optionally reduce R_a and/or R_b. O
Formula (I) according to claim 1, which may be converted to H or form a derivative thereof according to claim 1 and/or may form a pharmaceutically acceptable salt thereof. A method for producing a (S)-isomer of a compound or a derivative thereof. (7) Formula (VI) ▲ Contains mathematical formulas, chemical formulas, tables, etc. ▼ (VI) (wherein R_a and R_b are the same as above). (8) A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier. (9) The compound according to any one of claims 1 to 5, which is used for the treatment of viral infections. (10) Use of the compound according to any one of claims 1 to 5 in the manufacture of a drug used for the treatment of viral infections.