WO1999043689A1

WO1999043689A1 - Novel kdn derivatives and drugs containing the same as the active ingredient

Info

Publication number: WO1999043689A1
Application number: PCT/JP1999/000926
Authority: WO
Inventors: Kimio Furuhata; Haruki Yamada; Takayuki Nagai; Katsumi Ajisaka
Original assignee: Meiji Milk Products Co., Ltd.; The Kitasato Institute
Priority date: 1998-02-27
Filing date: 1999-02-26
Publication date: 1999-09-02
Also published as: AU2641599A

Abstract

KDN derivatives represented by general formula (1) or salts thereof and antiviral agents containing the same as the active ingredient wherein R1 represents lower alkyl, lower alkenyl or aralkyl; R?2 and R3¿ may be the same or different and each represents hydrogen or -SO¿3?H, provided that R?2 and R3¿ do not represent hydrogen atoms at the same time. These compounds are excellent in antiviral activity without showing any cytotoxicity.

Description

Description New KDN derivatives and pharmaceuticals containing them as active ingredients

Technical field

The present invention relates to a novel KDN derivative or a salt thereof useful for prevention and treatment of viral diseases such as influenza, and a medicament containing the same.

Background art

Influenza is a respiratory infection that spreads every year from winter to spring. When infected with the influenza virus, symptoms typically begin to appear after one to two days, such as runny nose, chills, fever, pain, malaise, and loss of appetite. These symptoms generally resolve in about a week, but in high-risk groups, such as the elderly and patients with underlying medical conditions, infants, and pregnant women, death often occurs with complications such as pneumonia and bronchitis. The danger of reaching is high.

Vaccination, a preventive measure against influenza infection, cannot be expected to be effective against new types of viruses. The anti-influenza drugs Aman Yujin and Liman Yujin give long-term side effects to the nervous system such as insomnia, dizziness and lack of concentration. The types have drawbacks such as being ineffective and the emergence of drug-resistant viruses. Recently, a substance that specifically inhibits sialidase (GG-167) (sialic acid derivative) (WO 91/1632 0). GS4 1 with a structure similar to GG-167 0 4 (US Pat. Nos. 5,763,483) have been developed and are undergoing extensive clinical trials. Sialidase is a spike-like glycoprotein that protrudes from the surface of virus particles. When the virus is released from infected cells, it breaks the bond between the sialidase and the receptor (sialic acid on the surface of infected cells), and separates them from the virus cells. Helps spread to cells It works and is an essential enzyme for the virus. Thus, GG-167 and GS410, specific inhibitors of sialidase, are expected to be effective as therapeutic and prophylactic agents against all types of influenza.

However, the development of anti-influenza drugs has only just begun. The potential of these sialidase inhibitors to inhibit resistant viruses and to inhibit host sialidase, which is involved in metabolic processes such as cell growth and plasma protein clearance, is currently unknown. For these reasons, there is a need for the development of anti-viral drug candidates that are expected to have even more new functions.

Disclosure of the invention

The present inventors have conducted intensive studies to develop a novel antiviral agent. As a result, a novel sialic acid derivative, 2-keto-3-dexoxy-D-glycerose-D-galacto-mononurosonic acid (KDN) The present inventors have found that the derivative or its salt has an excellent antiviral effect and does not have cytotoxicity, and thus completed the present invention.

That is, the present invention relates to the general formula (1)

[Wherein R ¹ is a lower alkyl group, indicates a lower alkenyl group or a lower Ararukiru group, R ² and R ³ shows the same or different and have good hydrogen atom or a group -S_〇 ₃ H (Here, R ² and R ³ are not both hydrogen atoms)

And a KDN derivative represented by the formula: or a salt thereof.

The present invention also provides a medicament comprising a KDN derivative represented by the above general formula (1) or a salt thereof as an active ingredient.

Furthermore, the present invention relates to a KDN derivative represented by the above general formula or a salt thereof and a pharmaceutically acceptable salt thereof.

Corrected It is intended to provide a pharmaceutical composition containing an acceptable carrier.

Furthermore, the present invention provides the use of a KDN derivative represented by the above general formula or a salt thereof as a medicament.

Furthermore, the present invention provides a method for treating a virus of a disease characterized by administering an effective amount of a KDN derivative represented by the above general formula or a salt thereof. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the anti-influenza virus activity (in vitro) for compound 2 and compound 3. FIG. 2 shows the toxicity of Compound 2 and Compound 3 on MDCK cells. FIG. 3 shows the therapeutic effect of compound 2 on influenza infection in mice. BEST MODE FOR CARRYING OUT THE INVENTION

The KDN derivative of the present invention is represented by the general formula (1). The lower alkyl group represented by ^1, is found include alkyl groups having 1 to 8 carbon atoms, specifically a methyl group, Echiru group, _n - propyl group, an isopropyl group, n - Bed butyl group, sec- butyl Group, tert-butyl group, n-pentyl group, n-hexyl group, n-heptyl group, n-octyl group, etc., of which methyl group, ethyl group, n-propyl group, n —Butyl groups are preferred.

Examples of the lower alkenyl group represented by R ¹ include an alkenyl group having 2 to 4 carbon atoms, specifically, a vinyl group, an aryl group, an isopropyl group, a 2-butenyl group, and a 1,3-butane group. A phenyl group, a 2-pentenyl group and the like are preferred, and an aryl group is particularly preferred.

The aralkyl group represented by R ¹ includes an aralkyl group having an alkyl moiety having 1 to 3 carbon atoms, and specifically, a phenylalkyl group such as a benzyl group, a phenyl group, and a 2-phenylpropyl group. Benzyl groups are particularly preferred. Good.

Examples of the salt of the KDN derivative (1) of the present invention include a base addition salt formed with respect to a free sulfonic acid group and a carboxyl group, for example, a salt with an alkali metal such as sodium, potassium and lithium. The ability to do so, sodium salt is particularly preferred.

The KDN derivative represented by the general formula (1) of the present invention can be synthesized by, for example, the following route using 3-dexoxy D-glycose mono-galacto-2-nonuloviranosonic acid as a starting material.

—

Process 1 4

[

[Reaction 1 4] Me Me

COOY

OR ¹ sulfate esterification

Deprotection ·

Hydrolysis

[Wherein, X represents a halogen atom (a chlorine atom, a bromine atom, etc.), Y represents a lower alkyl group (a methyl group, an ethyl group, etc.), and Z represents an acetyl group (an acetyl group, a benzoyl group, etc.). And M represents a hydrogen atom or an alkali metal atom (sodium, potassium, etc.), and R ¹ represents the same group as described above. ]

That is, compound (G) was synthesized in 6 steps from 3-Doxy-D-glycerol / S—D-galacto-2-nonuloviranosanoic acid (A) by reaction 1 and

By subjecting (G) to, for example, Reaction-1, Reaction-3, and Reaction-14, compounds (compound (I), compound (L), compound (Q)) included in the present invention can be synthesized. .

[Reaction 1]

Step 1 Step 1 is to convert 2-, 3-pi-2,3-dioxin-D-glycerone; 5-D-galact-1-2-nonuloviranosanoic acid (A) into a carboxylic acid ester, and then to the 4, 5, and 7-positions. This is the step of acylating the hydroxyl group of the compound to form a 0-acyl derivative, and further introducing a halogen atom at the 2-position. Esterification of the carboxylic acid is carried out by a known method usually known as an esterification reaction. When directly reacting with an alcohol, it is preferable to carry out the reaction in the presence of a dehydrating agent such as dicyclohexyl carboxylic acid (DCC), P-toluenesulfonic acid, and sulfuric acid. After the carboxylic acid is converted into a reactive derivative (eg, acid halide, acid anhydride, mixed acid anhydride, active ester, etc.), it is reacted with an alcohol, or the carboxylic acid is converted into an alkali metal salt (eg, sodium, potassium, etc.). , Cesium salts, etc.) and then reacting with an alkyl halide.

The acylation reaction can be carried out by any reaction usually used for acylation of a hydroxyl group. As the acylating reagent, for example, acyl chloride or acid anhydride is used, and the presence of a base such as N, N-dimethylaniline, N-methylmorpholine, dimethylamine, triethylamine, pyridine, sodium carbonate, carbonated lime, etc. The reaction can be carried out at −70 to 100 ° C. under or in the absence of. The introduction of a halogen atom is carried out when introducing a chlorine atom, for example, by saturating hydrogen chloride in an appropriate solvent or by adding hydrochloric acid in the presence of zinc chloride, and when introducing a bromine atom, introducing hydrogen bromide. It can be carried out by using an acid or a mixture thereof with sulfuric acid.

Step - 2, 2-position group of the compound (B) - introducing OR ^1, a reaction for producing 〇 Gurikoshido body (C).

Reaction, the compound (B) was dissolved in an alcohol corresponding to R ^1, sodium acetate, Natoriumu benzoic acid, the presence of a heavy metal salt catalyst such as silver triflate or silver oxide, stirred at room temperature ~ 5 0 ° C Can be performed.

Step-3 is a reaction for deacylation of compound (C) using an appropriate base to produce compound (D).

The base used here is not particularly limited as long as it does not affect other functional groups, but is preferably an alkali metal methoxide such as sodium methoxide and potassium methoxide. Examples of the solvent used include lower alcohols such as methanol and ethanol, and methanol is preferred.

The reaction temperature is usually from 10 to 50 ° C, and the reaction time is usually from 15 minutes to 10 hours, preferably from 1 to 2 hours.

Step 14 is a reaction for producing a compound (E) by introducing an isopropylidene group into the compound (D).

As the reagents used, there are acetone, 2-methoxypropene, and 2,2-dimethoxypropane, and 2,2-dimethoxypropane is preferable. Usually, an acid such as P-toluenesulfonic acid is used as a catalyst. .

Examples of the solvent used include non-protonic solvents such as acetone and N, N-dimethylformamide, and acetone is particularly preferred.

The reaction temperature is usually from 10 to 5 (TC, preferably from 10 to 30 ° C. The reaction time is usually from 15 minutes to 10 hours, preferably from 1 to 5 hours. is there.

Step-15 is a reaction for producing a compound (F) by acylating a hydroxyl group in the compound (E). This reaction can be carried out by a method similar to the acylation method shown in Step 11.

Step 16 is a reaction for removing the isopropylidene group of the compound (F) to produce the compound (G).

An acid is used in this reaction, and the acid is not particularly limited as long as it is used in a usual reaction, and inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, perchloric acid, and phosphoric acid are used. Examples thereof include acids or organic acids such as formic acid, acetic acid, oxalic acid, methanesulfonic acid, paratoluenesulfonic acid, trifluoroacetic acid, and trifluoromethanesulfonic acid, and acetic acid and trifluoroacetic acid are preferred.

The reaction temperature is usually 0 to 80 ° C, preferably 50 to 60 ° C. The reaction time is generally 15 minutes to 10 hours, preferably 1 to 5 hours.

[Reaction 1] Among K DN derivative or a salt thereof of the present invention, the compound R ² and R ³ are both represented by Kiichi S 0 ₃ H can child produced by the method shown in Reaction one 2.

That is, the hydroxyl group at the 8-position and 9-position of the compound (G) is sulfated to give a compound (H), and then the 4-, 5-, and 7-positions are silylated and the carboxylic acid ester is hydrolyzed. Compound (I) of the present invention can be synthesized.

[Reaction 1 3]

Among K DN derivative or a salt thereof of the present invention, the compound R ³ in R ² is Kiichi S_〇 ₃ H is hydrogen atom can be prepared by the method shown in Reaction one 3.

That is, the 9-position hydroxyl group of the compound (G) is protected by benzylation or the like to give a compound (J), and then the 8-position hydroxyl group is sulfated to give a compound (K). The compound (L) of the present invention can be synthesized by elimination of the protecting group, deacylation at the 4-, 5-, and 7-positions and hydrolysis of the carboxylic acid ester.

[Reaction 1 4]

Among the KDN derivatives or salts thereof of the present invention, the compound wherein R ² is a hydrogen atom and R ³ is a group —S 0 ₃ H can be produced by the method shown in Reaction 14.

That is, the hydroxyl group at the 9-position of the compound (G) is silylated to give a compound (M), and then the hydroxyl group at the 8-position is protected by, for example, benzylation to give a compound (N). Next, the 9-position silyl group is eliminated to give compound (0), and the 9-position hydroxyl group is sulfated to form a compound (P). Then, the 8-position hydroxyl protecting group is eliminated, and the 4- and 5-positions are removed. The compound (Q) of the present invention can be synthesized by performing deacylation at the 7-position and hydrolysis of the carboxylic acid ester.

For the protection of the hydroxyl group in the above reaction, any known protecting group can be used. For example, an acryl group (such as an acetyl group or a benzoyl group), an alkoxycarbonyl group (such as a methoxycarbonyl group or an ethoxycarbonyl group) can be used. And a benzyl group.

Hydroxy group silylation includes tert-butyldimethylsilyl group, tert-lo The introduction of a butyldiphenylurinyl group or the like can be mentioned. For example, in dimethylformamide, tert-butyldimethylsilyl halide is added in the presence of a base such as imidazole, triethylamine, and 4- (N, N-dimethylamino) pyridine. It can be carried out by reacting.

The removal of the protecting group for the hydroxyl group and the decomposition of the carboxylic acid ester can be carried out by known base hydrolysis. For example, the reaction can be performed by reacting with an aqueous alkaline solution such as an aqueous sodium hydroxide solution, an aqueous potassium hydroxide solution, or an aqueous sodium carbonate solution at room temperature to 5 (TC).

Sulfur esterification can be carried out by ordinary methods, and it is a mild sulfonation reagent known as sulfur trioxide / pyridine complex, sulfur trioxide / trimethylamine complex, sulfur trioxide / dimethylforma It is preferable to use a mid complex or the like. Examples of the reaction solvent include halogenated hydrocarbons such as methylene chloride, chloroform, carbon tetrachloride, and cyclobenzene; aromatic hydrocarbons such as benzene and toluene; ethers such as tetrahydrofuran and dioxane; acetonitrile; Aprotic polar solvents such as N, N-dimethylformamide can be used.

The reaction temperature is usually from 0 to 60 ° C, preferably from 10 to 30 ° C. The reaction time is generally 1 to 60 hours, preferably 24 to 48 hours.

Although the KDN derivative represented by the general formula (1) of the present invention is a sialic acid derivative, it does not have a sialidase inhibitory action such as GG-167 and GS410 as described above. For example, it is a novel compound having a completely new mechanism of action, such as exerting a therapeutic effect on influenza as described in the test examples herein. Therefore, the KDN derivative of the present invention is not limited to use as an anti-influenza drug. That is, a person skilled in the art would conceive of using the KDN derivative for drug screening for the treatment or prevention of viral diseases other than influenza, as well as for various other diseases, and an assay for implementing the method. The system can be easily set based on known techniques. Therefore, the present invention provides such a switch. The invention also encompasses a pharmaceutical use invention relating to a KDN derivative obtained by cleaning.

The KDN derivative of the present invention can be administered orally or parenterally (for example, intranasal administration) to animals and humans as it is or together with a conventional pharmaceutical carrier. In addition, the KDN derivative of the present invention may be used in various dosage forms, for example, orally administered drugs such as powders, granules, tablets, dragees, capsules, ampoules, etc., and pharmaceutical preparations such as subcutaneous, intramuscular or intravenous injections, suppositories, etc. It can be. These preparations include KDN derivatives alone or their derivatives and excipients, fillers, binders, wetting agents, disintegrants, surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, It can be produced by formulating in appropriate combination with carriers such as fragrances and coating agents.

The medicament of the present invention thus obtained varies depending on the age, weight, symptoms, and administration route of the patient, but is generally 0.6 to 30 Omg / day, preferably It is 5 to 20 Omg / day, and it is usually preferable to administer it in 3 to 4 times a day. Example

Hereinafter, the present invention will be specifically described with reference to Reference Examples and Examples, but the present invention is not limited to these.

Reference Example 1 Methyl 4,5,7,8,9-Penyl 1-Acetyl-1 2-Chloro-1,2,3-Dideoquin-1 D-Glycerol — D-Galacto-2-Nonuloviranosone

3D-Doxy-D-Glyceto 1 / 8-D-Galacto 2-Nonuloviranosonic acid (5.0 g, 18.6 g) is dissolved in methanol (100 m), and Dowex-50 (H +, 5 g) is added thereto. After stirring for 16 hours at, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography on silica gel (form: methanol = 10: 1). , Methyl ester derivative 4 g (yield 77 %) Was obtained as a colorless amorphous substance.

Acetic anhydride (30 mL) and pyridine (30 mL) were added to the methyl ester derivative (4. Og, 14), left at room temperature for 16 hours, and excess reagent was distilled off at 60 ° C under reduced pressure. The remaining oil was dissolved in ethyl acetate (50 m), washed twice with water (50 mL), and the ethyl acetate solution was dried over sodium sulfate, filtered, and concentrated to dryness to give a colorless powder, crude 6.86 g (yield 90%) of the hexane 0-acetyl derivative was obtained. Further, the crude derivative was purified by silica gel column chromatography (ether: hexane = 1: 1) to obtain 1.6 g of a colorless needle-like hex-0-acetyl derivative (80% yield) from an ether solution. %).

Hexa 0-acetyl derivative (2. Og, 3.74 Substrate 0 is dissolved in acetic acid (20 nl) and acetyl chloride (2 mL), and the solution is cooled to 0 ° C, saturated with dry hydrogen chloride, and cooled to room temperature. After leaving for 8 hours, the solvent was removed under reduced pressure at 30. A colorless powder was obtained, to which was added 30 mL of chloroform, washed twice with 20 mL of water, and dried over sodium sulfate. The thus obtained _c Glauber's salt was filtered and the solvent was distilled off to obtain the title compound (1.8 g, yield: 96%) as a colorless powder.

[] D ²² - 95.3. (c = 0.44, CHC s).

FAB-MS m / z: 511 (M + + l).

Anal. Calcd for C ₂₀ H ₂₇ 0 ₁₃ : C, 47.02; H, 5.33.

Found: C, 47.31; H, 5.45.

Reference Example 2 Methyl (benzyl 4,5,7,8,9-pentyl-cetyl-3-dexoxy-D-glycerol-D-galacto-2-nonuloviranoside)

Methyl 4,5,7,8,9-pentyl-0-acetyl-2, chloro-2,3-dideoxy-1-D-glycece-1 / S-D-galacto-2-nonuropyranoneto obtained in Reference Example 1 (10 g. 19.6 0 0 is dissolved in benzyl alcohol 50 mL, sodium benzoate (3.38 g, 23.5 瞧 0) is added, and the mixture is stirred at room temperature for 3 hours. The filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (n-hexane-ethyl acetate = 2: 1) and recrystallized from π-hexane-ethyl acetate to give the title compound as colorless prism crystals (7.5 g, 66 g). %).

mp 91-93 ° C

[H] _D ²⁵ + 0.57 ° (c = 1.04, CHC).

_{_{. Anal Calcd for C 27 H 34}} 0 14: C, 55.67; H, 5.88.

Found: C, 55.70; H, 5.9.

IRv _max ^{f ilm} cm- ^] : 1740 (00).

^] H-NMR (300MHz, CDC ₃ ) δ: 1.99 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

2.00, 2.01, 2.05, 2.10, 2.18 (each 3H, s, OAc),

2.73 (1H, dd, J = 4.5, 13.0 Hz, 3- Heq), 3.67 (3H, s, COOMe),

4.15 (1H, dd, J = 5.0, 12.0 Hz, 9—H),

4.20 (1H, dd, J = 2.0, 9.5 Hz, 6-H),

4.29 (1H, dd, J-3.0, 12.0 Hz, 9—H '), 4.46 (1H, d, J-12.0 Hz, — CHPh),

4.81 (1H, d, J = 12.0 Hz, — CH'Ph), 4.9 (1H, t 'J = 9.5 Hz, 5— H),

4.93 (1H, ddd, J = 4.5, 9.5, 11.5 Hz, 4-H),

5.36 (1H, dd, J = 2.0, 9.0 Hz, 7—H),

5.48 (1H, ddd, J = 3.0, 5.0, 9.0 Hz, 8—H),

7.27- 7.33 (5H, phenyl group).

Reference Example 3 Methyl (benzyl 3-dexoxy D-glycerol-H-D-galacto-2-nonuloviranoside)

Methyl obtained in Reference Example 2 (benzyl 4,5,7,8,9-penter 0-acetyl-13-doxy D-glycerol D-galacto-2-nonuloviranoside) netto (784 mg, 1.35 concealed 0 To the reaction mixture was added methanol (15mM), 28% sodium methoxydone ethanol solution (1mM) was added, and the mixture was stirred at room temperature for 1 hour.The dried Dowex-50 (H ⁺ , lg) was added to the reaction mixture. After summing, the resin is filtered and the resin is methanol I washed it. The filtrate and the washings were combined and evaporated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform-methanol = 10: 1) to give the title compound (461 mg, 92% ).

[H] _D ²⁵ -9.3 ° (c = 0.82, MeOH).

Anal. Calcd for dma H ₂₄ ₀₉ : C, 54.83; H, 6.50.

Found: C, 55.02; H, 6.46.

IR… ^fil H 50 (OH), 1720 (C = 0).

^{! H-NMR (300MHz, D} 2 0) <5: 1.81 (1H, dd, J = 11.5, 12.5 Hz, 3- Hax),

2.70 (1H, dd, J = 4.5, 12.5 Hz, 3-Heq), 3.58 (1H, t, J = 9.0 Hz, 540. 3.66 C1H, ddd, J = 4.5, 9.0, 11.5 Hz, 4-H),

3.72 (1H, dd, J = 4.0, 12.0 Hz, 9-H),

3.74 (1H, dd, J = 2.0, 4.0 Hz, 8—H), 3.79 (1H, d, J = 9.0 Hz, 6-H), 3.79 (3H, s, COOMe), 3.89 (1H, d, J = 2.0 Hz, 7-H),

3.92 (1H, d, J = 12.0 Hz, 9—H '), 4.60 (1H, d, J = 11.5 Hz,-CHPh),

4.81 (1H, d, J = 11.5 Hz,-CH'Ph), 7.37 to 7.7 (5H, phenyl group).

Reference Example 4 Methyl (benzyl 3-deoxy-1,8,9-diisopropylidene-D-glycemic mono-D-galacto-2-nonuloviranoside)

Acetone was added to methyl (benzyl 3-deoxy-D-glyceride-hi-ichi D-galacto-2-nonuloviranoside) nate (461 mg, 1.24 suloi) obtained in Reference Example 3.

(15 mL), 2,2-Dimethoxypropane (152 mg, 1.46 mg / l) and p-toluenesulfonic acid monohydrate (1 mg) were added, and the mixture was stirred at room temperature for 1 hour. The residue was neutralized with dry Dowex-1 (0H—, 1), the resin was removed by filtration, the resin was washed with acetone, and the filtrate and washings were combined and evaporated under reduced pressure. Purification by column chromatography (form: methanol-methanol: 10: 1) gave the title compound (460 mg, 90%) as a colorless powder.

[Shed] D ^{2 6} - 17.5. (C = 1.06, CHC ^ ₃ ). Anal. Calcd for C ₂₀ H ₂₈ ₀₉ : 58.24; H, 6.84.

Found: C, 58.50; H, 6.82.

IR _{max max} "'"' cur ¹ : 3350 (OH), 1740 (C = 0).

H NMR (300 MHz, D ₂₀ ) δ 1.39, 1.42 (each 3H, s, CMe ₂ ),

1.76 (1H, dd, J = 11.5, 12.5 Hz, 3-Hax),

2.64 (1H, dd, J = 4.5, 12.5 Hz, 3—Heq), 3.52 (1H, t, J = 9.0 Hz, 5-H), 3.8 (1H, ddd, J = 4.5, 9.0, 11.5 Hz, 4 -H),

3.65 (1H, dd, J = 1.0, 9.0 Hz, 6-H), 3.76 (3H, s, COOMe),

4.05 (1H, dd, J = 1.0, 6.0 Hz, 7—H),

4.05 (1H, dd, J = 6.0, 9.0 Hz, 9—H),

4.18 (1H, dd, J = 6.0, 9.0 Hz, 9-H,), 4.33 (1H, q, J = 6.0 Hz, 8-H), 4.54 (1H, d, J = 12.0 Hz, -CHPh), 4.75 (1H, d, J = 12.0 Hz,-CH'Ph), 7.32 to 7.40 (5H, phenyl group).

Reference Example 5 Methyl (benzyl 4, 5, 7-tree 0-cetyl-3-deoxy-1, 9-0-isopropylidene D-glycerol H-D-galacto 1-2-nuroviranoside)

Methyl (benzyl 3-deoxy-1,8,9-0-0-isopropyl pyridene-D-glycerol-D-galacto-2-nonuroviranoside) obtained in Reference Example 4

Pyridine (8nl) and acetic anhydride (5mL) were added to 460mg, 1.12, and stirred at room temperature for 16 hours. The reaction solution was evaporated under reduced pressure, water was added to the residue, and ethyl acetate (30ni) was added. The extracts were combined, washed with a saturated aqueous solution of sodium hydrogencarbonate (30 mL) and saturated saline (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure. Separation was performed by column chromatography (n-hexane monoacetate = 3: 1) to obtain the title compound (540 mg, 90%) as a colorless powder.

[H] D ²⁵ + 7.3 ° (c = 1.02, CHC s).

_{_{. Anal Calcd for C 2 sH 34}} 0 12: C, 57.99; H, 6.36. Found: C, 58.11: H, 6.32 ·

IR ^ _max ^{fi lra} cm— 1740 (00).

^J H NMR (300 MHz, D ₂₀ ) δ: 1.39, 1.42 (each 3H, s, CMe ₂ ),

1.76 (1H, dd, J = 11.5, 12.5 Hz, 3-Hax),

2.64 (1H, dd, J = 4.5, 12.5 Hz, 3-Heq), 3.52 (1H, t, J = 9.0 Hz, 5-H),

3.8 (1H, ddd, J = 4.5, 9.0, 11.5 Hz, 4—H),

3.65 (1H, dd, J = 1.0, 9.0 Hz, 6-H), 3.76 (3H, s, COO e),

4.05 (1H, dd, J = 1.0, 6.0 Hz, 7-H),

4.05 (1H, dd, J = 6.0, 9.0 Hz, 9—H),

4.18 (1H, dd, J = 6.0, 9.0 Hz, 9—H 'λ 4.33 (1H, q, J = 6.0 Hz, 8—H), 4.54 (1H, d, J = 12.0 Hz, -CHPh), 4.75 (1H, d, J = 12.0 Hz,-CH'Ph), 7.32-7.40 (5H, phenyl group).

Reference Example 6 Methyl (benzyl 4,5,7-tri-0-acetyl-3-dexoxy-D-glycemic mono-D-galacto-2-nonuloviranoside)

Methyl (benzyl 4,5,7-tree 0-acetyl-3-dexoxy 8,9-0-isopropylidene-D-glycerol-Hi-D-galacto-2-nonuloviranoside) nate (540 mg, 1.00) obtained in Reference Example 5 To the mixture, 10 mL of a 90% acetic acid solution was added, and the mixture was stirred for 2 hours at 60 ° C. After the reaction solution was distilled off under reduced pressure, the residue was subjected to silica gel column chromatography (form: methanol: 100: 1 in chloroform). ) To give the title compound (390 mg, 78%) as a colorless powder.

[H] D ²⁵ + 11.2 ° (c = 0.72, CHC s).

_{_{. Anal Calcd for C 23 H 3O}} 0 12: C, 55.42; H, 6.07.

Found: C, 55.51: H, 6.04.

IR _raax ""'cm— ¹ : 3400 (OH), 1720 (C = 0).

1 H-NMR (300MHz, CDC ₃ ) o: 1.99, 2.02, 22.12 (each 3H, s, OAc),

2.06 (1H, dd, J = 11.5, 13.0 Hz, 3— Hax), 2.39 (1H.dd, J = 5.0, 9.0 Hz, 9-0H),

2.87 (1H, dd, J = 5.0, 13.0 Hz, 3-Heq),

3.51 (1H, dt, J = 5.0 '12.0 Hz, 9-H),

3.68 (1H, ddd, J = 3.0, 9.0, 12.0 Hz, 9-H '), 3.79 (3H, s, COOMe),

3.89 (1H, d, J = 5.0 Hz, 9-0H),

3.99 (1H.ddd, J = 3.0, 5.0, 9.0 Hz, 8—H),

4.01 (1H, dd, J = 2.0, 9.5 Hz, 6-H), 4.49 (1H, d, J = 12.0 Hz, one CHPh), 4.82 (1H, d, J = 12.0 Hz, -CH 'Ph), 4.89 (1H, t, J = 9.0 Hz, 5—H),

4.94 (1H, ddd, J = 5.0, 9.5, 11.5 Hz, 4—H). 5.05 (1H, t, J = 9.5 Hz, 5—H). 5.05 (1H, dd, J = 2.0, 9.0 Hz, 7 -H). 7.28 to 7.38 C5H, phenyl group). Example 1 Trisodium (benzyl 3-deoxy 8, 9-di-0-s-fo-D-glycerol-Hi-D-galacto-2-nonuroviranoside) nate (compound Synthesis of product 1)

(1) Methyl (benzyl 4,5,7-tree 0-acetyl-8,9-di-0-sulfo 3-deoxy D-glycerol H-D-galacto-2-nonuropyranoside) nate triethylamine

Methyl (benzyl 4,5,7-tri-0-acetyl-3-dexoxy D-glycerol H-D-galacto 2-nonuloviranoside) netrate (100 mg. Powder Molecular 'Sieves 4A (200mg) and zirconium trioxide / trimethyl complex (32mg, 0.33ml) were added, and dried DMF was added.

(lmL), and the mixture was stirred at room temperature under an argon stream for 40 hours. Triethylamine (1 mL) was added to the reaction solution, and the mixture was filtered and evaporated under reduced pressure at 6 (TC. Preparative TLC

(Toluene-ethanol-triethylamine = 3: 1: 0.5), and the residue was extracted with chloroform-ethanol (2: 1) and dried to dryness to give the title compound (85 mg, 85 mg, 85%) as a colorless powder. %).

[H] D ²⁰ + 2.8 ° (c = 1.0, MeOH). Anal. Calcd for C ₂₃ H ₃₀ 0 ₁₆ S ₂ -CeHsoN: C, 50.71; H, 7.30; N, 3.38.

Found: C, 50.65; H, 7.36; N, 3.28.

IR ai 'n'cnf ¹ : 1745 (C = 0), 1600 (C = 0, 810 (S-0).

^] H-NMR (300MHz, CDC ₃ ) δ: 1.96, 1.98, 2.06, (each 3H, s, OAc),

1.97 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

2.74 (1H, dd, J = 5.5, 13.0 Hz, 3-Heq), 3.80 (3H, s, COOMe),

4.07 (1H, dd, J = 6,5, 11.0 Hz, 9-H),

4.26 (1H, dd, J = 2.0, 10,5 Hz, 6-H), 4.60 (1H, d, J = 11.5 Hz,-CHPh), 4.61 (1H, dd, J = 2.5, 11.0 Hz, 9_H ' ), 4.91 (1H, t, J = 9.5 Hz, 5—H), 4.92 (1H, d, J = 11.5 Hz,-CH'Ph),

4.97 (1H, ddd, J = 6.5, 2.5, 4.0 Hz, 8—H),

5.03 (1H, ddd, J = 5.5, 9.5, 11. Hz, 4-H),

5.61 (1H, dd, J = 2.0, 4.0 Hz, 7_H), 7.30 to 7.36 (5H, phenyl group), 1.35 (18H, t, J-7.5 Hz, NEt ₃ ), 3.15 (12H, q, J = 7.0 Hz, NEt ₃ ).

(2) Methyl obtained in (1) (benzyl 4,5,7-tri-〇-acetyl-1, 9-zy 0-sulfo 3-deoquin-D-glycerol D-galactose 2-nonuloviranoside) To the solution was added 0.2N aqueous sodium hydroxide solution (4 mL) to triethylamine (40 mg, 0.048 mmol), and the mixture was stirred at room temperature for 6 hours.The reaction mixture was neutralized with Dowex-50 (H ⁺ , 0.5 g), and the resin was neutralized. The filtrate and the washings were combined, washed with ether (10 mL) three times, and the aqueous layer was freeze-dried.The remaining colorless powder was dissolved in water (2 mL). After passing through a column of Dowex-50 (Na +, 5 g) and washing with water, the combined filtrates were lyophilized, and the residue was dissolved in water (3 m) and dissolved in a Millipore filter ( After purification by lyophilization, a colorless powdery compound 1 (23 mg, 97%) was obtained.

[Shed] _D ^2. -18.0. (c = 0.4, H ₂ 0).

Anal. Calcd for Ci eHj ₉ 0i ₃ S ₂ Na ₃ : C, 34.79; H, 3.47. Found: C, 34.65; H, 7.36.

FAB-MS m / z: 553 (M ⁺ +1).

IR level _max fi '-' cm- ¹ : 1745 (C = 0), 1605 (C = C), 810 (S-0)

] H - discussions _{R (300MHz, D 2 0)} δ: 1.60 (ΙΗ 't, J = 12.0 Hz, 3- Hax),

2.62 (1H, dd, J-4.5, 12.0 Hz, 3-Heq), 3.47 (1H, t, J = 9.0 Hz, 5—H),

3.61 (1H, ddd, J = 9.0, 4.5, 12.0 Hz, 41-H),

3.86 (1H, dd, J = 3.0, 13.0 Hz, 9—H),

4.00 (1H, dd, J = 1.0, 9.5 Hz, 6-H),

4.06 (1H, dd, J = 3.0, 13.0 Hz, 9—H '),

4.48 (1H, dd, J = 1.0. 3.0 Hz. 7-H), 4.49 (1H, d, J = 10.5 Hz, — CH— Ph), 4.77 (1H, d, J = 10.5 Hz,-CH- Ph) ), 4.73-4.79 (2H, m, 8, 9-H),

7.40 (5H, m, Ph).

Example 2 Synthesis of disodium (benzyl 3-deoxy 8-0-sulfo D-glycerol-1-D-galacto-2-nonu α-pyranoside) nate (compound 2)

(1) Methyl (benzyl 4,5,7-tri-1 0-acetyl-9-0-benzoyl-3-dexoxy D-glyceric α-D-galacto-2-nonuloviranos)

5 ml of methyl (benzyl 4,5,7-tri- ァ -acetyl-3-doxy-D-glyceichi-1-D-galacto-2-nonuloviranoside) obtained in Reference Example 6 was added to pyridine (140 mg, 0.28 mmol) in 5 mL of pyridine. To dissolve and add benzoic anhydride 5 3 8 mg

(3.38 mmoi) and 1 mg of 4-dimethylaminopyridine were added, and the mixture was stirred at room temperature for 16 hours. 3 OmL of saturated sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted three times with chloroform. The extracts were combined, washed with 3 OmL of saturated saline, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure. Silica gel column chromatography of the residue

(149 mg, 88%) of the title compound (149 mg, 88%) as a colorless powder. Obtained.

[H] D ²⁴ + 12.6 ° (c-0.99, CHC ^ ₃ ).

_{_{. Anal Calcd for C 30 H 34}} 0 13: C, 59.80; H, 5.69.

Found: C, 59.96: H, 5.58.

IR .ax '^ -cm- ¹ : 3450 (OH), 1720CC = 0), 1600 (C = 0.

^J H-NMR (300MHz, CDC) 1.99, 2.02, 2.09 (each 3H.s, OAc),

2.09C1H, dd, J = 11.5, 13.0Hz, 3- Hax),

2.87 (1H, dd, J = 5.0, 13.0Hz, 3-Heq), 3.79 (3H, s, COOMe),

3.96C1H, br d, J = 3.0Hz, 9-H),

4.08 (1H, dd, J = 2.0, 9.0Hz, 6-H),

4.28 (2H, m, 8- and 9-H '), 4.5K1H, d, J = 12.0Hz,-CHPh),

4.84C1H, d, J = 12.0Hz,-CH'Ph),

4.94 (1H, ddd, J = 5.0, 9.0, 11.5Hz, 4-H),

5.0K1H, t, J = 9.0Hz, 5-H),

5.27 (1H, dd, J = 2.0, 8.0Hz, 7-H),

7.31-8.1K10H, phenyl group).

(2) Methyl (benzyl 4,5,7-tri-1 0-acetyl-1 9-1 0-benzoyl-1 8-0-sulfo-3-dexoxy D-glyceichi-1 D-galacto-2-nonuroviranoside) Nate '' Trietil

Methyl (benzyl 4,5,7-tree 0-acetyl-91-0-benzoyl-1 3-deoxy D-glyceic D-galacto-2-nonulopyranoside) nate (40 mg) obtained in (1) , 0.066 fractions 0), powdered molecular sieves 4M 200 mg) and trimethylamine trioxide (18 mg, 0.166 hidden), dry DMF (ImL) was added, and the mixture was stirred at room temperature under an argon stream for 40 hours. Stirred. Triethylamine (ImL) was added to the reaction solution, and the mixture was filtered and evaporated under reduced pressure at 60 ° C. The residue was purified by Preparative TLC (toluene-ethanol-triethylamine = 3: 1: 0.5). Purification gave the title compound as a colorless powder (42 mg, 81%).

[H] D ²⁴ ten 11.4. (c = 1.11, CHC ₃ ).

Anal. Calcd for C ₃₆ H ₄₃ 0 ₁₆ NS: C, 55.16; H, 6.30; N, 1.79.

Found: C, 55.23; H, 6.19; N, 1.88.

IR _max ^{ii lm} cm ~ ^l : 1720 (C = 0), 1600 (C = 0, 810 (S-0).

] H-Oki (300MHz, CDC ₃ ) δ: 1.99, 2.00, 2.04 (each 3H, s, OAc),

2.01 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

2.73 (1H, dd, J = 5.0, 13.0 Hz, 3-Heq), 3.84 (3H, s, COOMe),

4.34 (1H, dd, J = 5.0, 12.0 Hz, 9-H),

4.41 (1H, dd, J = 2.0, 9.5 Hz, 6-H), 4.64 (1H, d, J = 12.0 Hz, -CHPh, 4.93 (1H, t, J = 9.5 Hz, 5-H), 4.94 ( 1H, d, J = 12.0 Hz. — CH'Ph),

5.02 (1H, dd, J = 3., 12.0 Hz, 9—H '),

5.11 (1H, ddd, J = 5.0, 9.5, 11.5 Hz, 4—H),

5.13 (1H, dt, J = 3.0, 5.0 Hz, 8—H), 5.66 (1H, dd, J = 2.0, 5.0 Hz, 7-H), 7.29 to 8.10 (10H, phenyl group), 1.25 (9H, t, J = 7.5 Hz, NEt ₃ ),

3.05 (6H, q, J = 7.0 Hz, NEt ₃ ).

(3) Methyl (benzyl 4,5,7-tree 0-acetyl-9-0-benzoyl-1 8-0-sulfo 3-dexoxy D-glycerol H-1 D-galactone 2-) obtained in (2) Nonuroviranoside) nate triethylamine (40 mg, 0.048), 4 mL of a 0.2N aqueous sodium hydroxide solution was added, and a colorless powdery compound 2 (23 mg, 97%) was obtained in the same manner as in Example 1. Was.

[] D ²⁰ -19.7 ° (c = 0.73, H2O).

Anal. Calcd for C ₁₆ H ₂ . 0 ₁₂ SNa ₂ : C, 44.25: H, 4.64.

Found: C, 44.65; H, 4.36. FAB-MS m / z: 435 (M ++ 1).

IR _raax f i'-cnf ¹ : 1740 (00), 1605 (C = C), 810 (S-0).

'H-NMR (300MHz, D 2 0) δ 1.60 (1H, t, J = 12.0 Hz, 3- Hax), 2.65 (1H, dd, J = 4.5, 12.0 Hz, 3-Heq), 3.46 (1H, t, J = 9.5 Hz, 5-H), 3.59 (1H, ddd, J = 9.0, 4.5, 12.0 Hz, 4 -H),

3.83 (IE dd, J = 1.0, 12.5 Hz, 940,

3.88 (1H, dd, J = 1.0, 9.5 Hz, 6-H),

4.27 (1H, dd, J = 3.5, 12.5 Hz, 9-H '),

4.47 (1H, dd, J = 1.0, 2.5 Hz, 7-H), 4.49 (1H, d, J = 10.5 Hz,-CH-Ph),

4.55 (1H, ddd, J = 2.5, 6.5, 3.5 Hz, 8-H),

4.72 (1H, d, J = 10.5 Hz,-CH-Ph), 7.40 (5H, m, Ph).

Example 3 Synthesis of disodium (benzyl 3-deoxy-9-0-0-sulfo D-glycerol H-D-galacto-2-nonuloviranoside) nate (compound 3)

(1) Methyl (benzyl 4,5,7-tri-O-acetyl- 9-〇-t-butyldimethylsilyl-13-dexoxy-D-glycemic-D-galacto-2-nonuloviranoside)

Methyl (benzyl 4,5,7-tree-acetyl-3-deoxy-D-glycethone-1-galatato-2-nonuloviranoside) net obtained in Reference Example 6 (220 mg, 0.44 mmoi) was passed through an argon stream. Medium DMF (2mL) was added and dissolved, cooled to 0 ° C, imidazole (120mg, 1.77moi), tert-butyldimethylsilyl chloride

(133 mg, 0.88 moi) was added, and the mixture was stirred at room temperature for 16 hours. Ice water was added to the reaction solution, and extracted three times with ether (30 mL). The extracts were combined, washed with water, dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated under reduced pressure. The residue was purified using silica gel column chromatography (n-hexane-ethyl acetate = 2: 1) to give the title compound (162 mg, G0%) as a colorless powder.

[H] D ² + 5.2 ° (c = 0.84, CHC s).

Anal. Calcd for C ₂₉ H ₄₄ 0 ₁₂ Si: C, 56.84: H, 7.24.

Found: C, 57.09; H, 7.18. IR _rarax ¹ !!! — ¹ : 3450 (OH), 1750 (C = 0).

Ή-NMR (300MHz, CDC ₃ ) ό: 0.07 (6H, s, Si e ₂ ),

0.90 (9H, s, SiC e ₃ ), 1.99, 2.01, 2.08 (each 3H, s, OAc),

2.05 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

2.83 (1H, dd, J = 5.0, 13.0 Hz, 3-Heq),

3.45 (1H, d, J-5.0 Hz, 8-0H), 3.61 (1H, dd, J = 6.0, 11.0 Hz, 9-H),

3.70 (1H, dd, J = 3.5, 11.0 Hz, 9-H '), 3.78 (3H, s, COOMe),

4.02 (1H, m, 8-H), 4.11 (1H, dd, J = 2.0, 10.0 Hz, 6-H),

4.51 (1H, d, J = 12.0 Hz,-CHPh), 4.84 (1H, d, J = 12.0 Hz,-CH'Ph),

4.95 (2H, m, 4-H, 5-H), 5.13 (1H, dd, J = 2.0, 8.5 Hz, 7-H),

7.30 to 7.38 (5H, phenyl group).

(2) Methyl (benzyl 4,5,7-tree 0-acetyl- 8-0-ben-byl 9-0-t-butyldimethylsilyl-3-deoxy-D-glycerol-Hi-D-galacto-2-nonuroviranone) Nate

Methyl (benzyl 4,5,7-triacetyl-9-0-t-butyldimethylsilyl-13-dexoxy-D-glycerol-H-D-galacto-2-nonuloviranoside) nate obtained in (1) (162 mg) Was dissolved in pyridine (15), benzoyl chloride (0.5 mL) was added, and the mixture was stirred at room temperature for 6 hours, ice water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (30 mL). The extract was washed with a saturated aqueous solution of sodium hydrogencarbonate and saturated saline, dried over anhydrous sodium sulfate, filtered, and the filtrate was distilled off under reduced pressure.The residue was subjected to silica gel column chromatography (n-hexane-ethyl acetate = 2 : 1) to give the title compound (148 mg, 78%) as a colorless powder.

[H] D ² † 10.3 ° (c = 0.99, CHC ₃ ).

Anal.Calcd for C ₃₆ H ₄₈ 0 ₁₃ Si: C, 60.32; H, 6.75.

Found: 60.44; H, 6.69. IR: ^fi ' ^m cm-]: 1740 (C = 0), 1590, 1570 (OC).

^J H-NMR (300MHz, CDC s) δ -0.04 (6H, s, SiMe ₂ ), 0.83 (9H, s, SiCMe ₃ ).

1.99, 2.04, 2.12 (each 3H, s, OAc),

1.99 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

.69 (1H, dd, J = 5.0, 13.0 Hz, 3-Heq),

3.29 (3H, s, COOMe), 3.76 (1H, dt, J = 2.0, 11.0 Hz, 9-H),

3.96 (1H, br d, J = 11.0 Hz, 9-H '), 4.02 (1H, m, 8—H),

4.23 (1H, br d, J = 9.5 Hz, 6—H), 4.50 (1H, d, J = 12.0 Hz,-CHPh),

4.89 (1H, d, J = 12.0 Hz, — CH'Ph), 4.91 (1H, t, J = 9.5 Hz, 5-H),

5.00 (1H, ddd, J = 5.0, 9.5, 11.5 Hz, 4-H), 5.58 (2H, m, 7-H, 8-H),

7.31 to 8.09 (10H, phenyl group).

(3) Methyl (benzyl 4, 5, 7—tree 0—acetyl-8—0—benzoyl-3-deoxy—D—glyceic monosaccharide D—galacto 2-nonuroviranoside

Methyl obtained in (2) (benzyl 4,5,7-tri-0-acetyl-0-benzoyl-910-t-butyldimethylsilyl 3-dexoxy-D-glycerol-D-galacto To 2-nonuroviranoside) nate (148 mg, 0.21 mraof), 10% of a 90% acetic acid aqueous solution was added, and the mixture was stirred at 60 ° C for 6 hours. After evaporating the reaction solution under reduced pressure, the residue was purified by silica gel column chromatography (form:

Purification using 100: 1) afforded the title compound (86 mg, 69).

[H] D ²⁵ -5.1 ° (c = 1.02, CHC).

Anal. Calcd for C _3. H ₃₄ 0 _{) 3} : 59.80: H, 5.69.

Found: C, 60, 03; H. 5.62.

IR ^iiln 'cnr]: 3500 (OH), 1740 (00), 1590, 1570 (C = C).

^! H-NMR (300MHz, CDC) 6 1.99 (1H, dd, J = 11.5, 12.5 Hz, 3-Hax),

2.00, 2.05, 2.20 (each 3H, s, OAc), 2.60 (1H, dd, J = 5.5, 9.0 Hz, 9-OH). 2.72 (1H, dd, J = 5.0, 12.5 Hz, 3- Heq), 3.24 (3H, s, COO e),

3.66 (1H, ddd, J = 3.0, 5.5, 13.0 Hz, 9-H),

3.95 (1H, ddd, J = 3.0, 9.0, 13.0 Hz, 9- H '),

4.26 (1H, dd, J = 2.0, 9.5 Hz, 6-H), 4.49 (1H, d, J = 12.0 Hz,-CHPh),

4.85 (1H, d, J = 12.0 Hz,-CH'Ph),

5.00 (1H, ddd, J = 5.0, 9.5, 11.5 Hz, 4-H),

5.08 (1H, t, J = 9.5 Hz, 5-H), 5.41 (1H, dd, J = 2.0, 9.0 Hz, 7_H), 5.49 (1H, dt, J = 3.0, 9.0 Hz, 8-H), 7.31 to 8.11 (wei phenyl group).

(4) Methyl (benzyl 4,5,7-tree 0-acetyl- 8-0-benzoyl 9-1 0-sulfo 3-doxy-I-D-glycose mono-D-galacto-2-no-nuroviranoside) N

Methyl (benzyl 4,5,7-tri-II-cetyl-8-0-benzoyl-1-3-dexoxy D-glycerol-1-D-galacto-2-nonuroviranoside) nate obtained in (3) (40 mg, 0.066 oi), powdered molecular 'Sieves 4A (200 mg) and trimethylamine complex (23 mg, 0.166) were added, and the reaction was carried out in the same manner as in the synthesis of Example 2. This gave the title compound as a powder (40 mg, 77%).

[Shed] D ²⁴ - 7.2. (C = 1.04, CHC s).

Anal. Calcd for C _3S H ₄₉ 0) ₆ NS: 55.16; H, 6.30; N, 1.79.

Found: 55.23; H, 6.19; N, 1.88.

IR v _raax ^{ri lm} cnf 1740 (OH), 1590, 1570 (C = C), 810 (S-0).

Ή-NMR (300MHz, CDC s) 6: 1.98, 2.00, 2.12 (each 3H, s, OAc),

1.98 (1H, dd, J = 11.5, 13.0 Hz, 3-Hax),

2.71 (1H, dd, J = 5.0, 13.0 Hz, 3-Heq), 3.39 (3H, s, COOMe),

4.23 (1H, dd, J = 2.0, 9.0 Hz, 6-H), 4.27 (1H, dd, J = 5.0, 12.0 Hz, 9-H),

4.49 (1H, dd, J = 2.0, 12.Hz, 9-H '), 4.52 (1H, d, J = 12.0 Hz, -CHPh), 4.90 (1H, d, J = 12.0 Hz,-CH'Ph), 4.92 (1H, t, J = 9.0 Hz, 5-H),

4.96 (1H, ddd, J = 5.0, 9.0, 11.5 Hz, 4—H),

5.55 (1H, dd, J = 2.0, 7.0 Hz, 7—H),

5.78 (1H, dd, J = 2.0, 5.0, 7.0 Hz, 8-H),

7.30 to 8.10 (10H, phenyl group), 1.25 (9H, t, J = 7.5 Hz, NEt ₃ ).

3.03 (6H, q, J-7.0 Hz, NEt ₃ ).

(5) Methyl (benzyl 4,5,7-tree 0-acetyl-1-8-0-benzoyl-1-9-11-1 sulfo 3-dexoxy D-glyce α-hi-1 D-galact-1 obtained in (4) 2-Nonuloviranoside) nate ■ To a solution of triethylamine (40 mg, 0.048 mmoi) in 0.2 N aqueous sodium hydroxide (4 mL), and in the same manner as in Example 1, colorless powdered compound 3 (23 mg, 97%) I got

[] D ²⁰ -33.2 ° (c = 0.67, H ₂ 0).

_{_{. Anal Calcd for Ci 6 H 20}} Oi 2 SNa 2: C, 44.25; H, 4.64.

Found: C, 44.65; H, 4.36.

FAB-MS m / z: 435 (M + 11).

_{^{IR ma x fi lra cm- ':}} 1740 (C = 0), 1605 (OC), 810 (S- 0).

- Tsuyoshi _{R (300MHz, D 2 0)} o: 1.64 (1H, t, J = 12.0 Hz, 3- Hax),

2.71 (1H, dd, J = 4.5, 12.5 Hz, 3-Heq), 3.52 (1H, t, J = 9.5 Hz, 5-H,

3.59 (1H, ddd, J = 9.0, 4.5, 12.0 Hz, 4-H),

4.19 (1H, dr.dt, J = 2.5, 11.5 Hz, 9-H),

3.71 (1H, dr.d, J = 9.5 Hz, 6—H), 3.93 (2H, br.s, 7-H, 8_H),

4.30 (1H, dr.d, J = 11.5 Hz, 9-H '), 4.51 (1H, d, J = 11.0 Hz, -CH-PhX

4.71 (1H, d, J = 11.0 Hz,-CH-Ph), 7.40 (5H, m, Ph).

Test Example 1 Anti-influenza virus activity (in vitro)

Compound 2 (Example 2) and Compound 3 (Example 3) were dissolved in dimethyl sulfoxide (DMSO), and diluted with water so that the final concentration of DMSO was 1% (ν / ν). I shouted. On the other hand, the positive control, ribulin, was dissolved in water. As a negative control, DMSO diluted with water to a final concentration of 1% (ν / ν) was used. 4 8 we 11 MDCK (Madin-Darby canine kidney) cells (JM Woods, et al., Antimicrob.Agents Chemother., 37, 1473-1479 (1993)) cultured in culture plate, influenza in the presence of trypsin The virus AZPR / 8/34 strain was infected with 0.001 MOI (multiplicity of infection), and at the same time, Compound 2, Compound 3, Ribavirin and DMSO solution were added. This 5% C_〇 ₂ conditions, and cultured for 3 days at 37 ° C. Since cells infected and proliferated by the influenza virus die, the cell viability is measured by the MTT method as an indicator of anti-influenza virus activity (R. Pauels, et al., J. Virol. Methods, 20, 309-321 ( 1988)). The results are shown in Figure 1. The following is clear from FIG.

When only DMSO was added, the cell viability was about 40%. In contrast, when Compound 2 or Compound 3 was added at a concentration of 95 g / m, it was confirmed that the cells were almost 100% viable. This value was almost the same as when ribavirin was added at a concentration of 22 g / m. From these results, Compound 2 and Compound 3 of the present invention are useful as anti-influenza virus agents.

Test Example 2 Cytotoxicity

As in Test Example 1, Compound 2 (Example 2), Compound 3 (Example 3), Ribavirin or DMSO solution was added to influenza virus-uninfected MDCK cells cultured in a 48-well 1 culture plate. These 5% C_〇 ₂ conditions, and cultured for 3 days at 37 ° C, and the cell viability was measured by MTT method. The result is shown in figure 2. Figure 2 shows that cells containing Compound 2 or Compound 3 have almost 100% cell survival and extremely low cytotoxicity.

Test example 3 Experimental treatment for mouse infection

Mouse-adapted influenza virus A / PR / 8/34 stock solution (50% chicken eggs

2 8 Corrected form (Rule 91 Infective dose = 1 0 ^{^8.5)} was diluted to 1 0 ⁴ times with phosphate buffered saline (PBS) containing a 0.1% © shea serum albumin to prepare a virus dilutions. BALB / c mouse (female, 7 weeks old) (Japan SLC) was anesthetized by intraperitoneal administration of 0.2 mL of pentobarbi sodium solution (5 mL of normal saline, 1 mL) and 10 mL of virus diluent was administered. Infection was achieved by nasal inoculation. Compound 2 (Example 2) was used as a physiological saline solution (1 mgZniL), and administered twice a day for 4 days before, 4 hours after and 4 days after virus inoculation under pentobarbital anesthesia at a dose of lmgZkg per day. The mice were administered intranasally. For the control group, physiological saline was intranasally administered instead of the compound 2 solution. The survival rate of the mice was observed for 21 days after infection with the influenza virus, and the in vivo anti-inffluenza virus activity of Compound 2 was evaluated by comparison with the control group. The results are shown in Figure 3. The following is clear from FIG. The mice in the control group which received saline intranasally began to die on day 8 of virus inoculation, and the survival rate from day 9 to day 21 was 40%. In contrast, mice administered compound 2 nasally had a 70% survival rate from day 8 to day 18, and 60% of mice survived on day 21. In comparison, increased survival rates and prolonged survival days were observed. From these results, compound 2 has an excellent therapeutic effect on influenza infection and is useful as an antiviral agent.

Formulation Example 1 Compound 2 (Example 2) is 10% (W / W), benzalkonium chloride is 0.04% (W / W), phenylethyl alcohol is 0.40% (W / W), Prepare an aqueous solution so that the purified water is 89.56% (W / W).

Formulation Example 2 Compound 2 (Example 2) is 10% (W / W), benzalkonium chloride is 0.04% (W / W), polyethylene glycol 400 is 10.0% (W / W) An aqueous solution is prepared so that propylene glycol is 30% (WW) and purified water is 39.96% (WZW).

Formulation Example 3 A dry powder is prepared so that compound 2 (Example 2) is 40% (W / W) and lactose is 60% (W / W). Formulation Example 4 Compound 2 (Example 2) Air port so that force is 0% (WZW), lecithin is 0.5% (W / W), freon 11 is 34.5%, freon 12 is 55%. Prepare a sol. Industrial applicability

INDUSTRIAL APPLICABILITY The novel KDN derivative of the present invention has excellent antiviral activity and low toxicity, and thus is useful as a therapeutic or preventive agent for viral infections, especially influenza infections.

Claims

[Wherein, R ′ represents a lower alkyloxy p group, a lower alkenyl group or an aralkyl group, and R ² and R ³ may be the same or different and each represent a hydrogen atom or a group S 0 ₃ H (provided that R ² And R ² are not simultaneously hydrogen atoms)

Or a salt thereof.

Enclosure

2. The KDN derivative according to claim ^1, wherein R ^{1 is a} benzyl group.

3. A medicament comprising the KDN derivative according to claim 1 or 2 or a salt thereof as an active ingredient.

4. The medicament according to claim 3, which is a therapeutic agent for a viral disease.

5. A pharmaceutical composition comprising the KDN derivative according to claim 1 or 2, or a salt thereof, and a pharmaceutically acceptable carrier.

6. Use of the KDN derivative or a salt thereof according to claim 1 or 2 as a medicine.

7. The use according to claim 6, wherein the medicament is a remedy for diseases of children.

8. A method for treating a viral disease, which comprises administering an effective amount of the KDN derivative or a salt thereof according to claim 1 or 2.