WO1999043690A1 - L-4'-arabinofuranonucleoside compound and medicine composition comprising the same - Google Patents

L-4'-arabinofuranonucleoside compound and medicine composition comprising the same Download PDF

Info

Publication number
WO1999043690A1
WO1999043690A1 PCT/JP1999/000827 JP9900827W WO9943690A1 WO 1999043690 A1 WO1999043690 A1 WO 1999043690A1 JP 9900827 W JP9900827 W JP 9900827W WO 9943690 A1 WO9943690 A1 WO 9943690A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
compound
formula
nucleoside
compound represented
Prior art date
Application number
PCT/JP1999/000827
Other languages
French (fr)
Japanese (ja)
Inventor
Hiroshi Sato
Yuichi Yoshimura
Noriyuki Ashida
Kenji Sudo
Tomoyuki Yokota
Original Assignee
Rational Drug Design Laboratories
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rational Drug Design Laboratories filed Critical Rational Drug Design Laboratories
Publication of WO1999043690A1 publication Critical patent/WO1999043690A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom

Definitions

  • the present invention relates to an L-4′-thioarabinofuranonucleoside compound and a pharmaceutical composition containing the compound as an activity, particularly to an anti-hepatitis virus agent.
  • L-nucleosides having an unnatural structure have been shown to have an anti-viral effect, and are attracting attention due to their low toxicity. For example, it has been reported that 2'-fluoro-5-methyl-13-L-arabinofuranosylperacil (L-FMAU) has anti-B virus (HBV) activity and low cytotoxicity.
  • L-FMAU 2'-fluoro-5-methyl-13-L-arabinofuranosylperacil
  • HBV anti-B virus
  • Antiviral Chemistry & Chemotherapy (1995), 6 (3), 138-142 include a-L-arabinofuranosylcytosine, ⁇ -L-xylofuranosylcytosine and Although some anti-HCMV (human cytomegalovirus) activity was confirmed in one L-anomer of a-L-FMAU, anti-H CMV was observed in other ⁇ -L anomers and ⁇ -L anomers of the above compounds. No activity has been confirmed.
  • Japanese Patent Publication No. 8-504753 discloses that 2 ', 3'-didehydro-2', 3'-dideoxy-14'-thio- ⁇ -l-cytidine and 2 ', 3'didehydro 2', 3 ' The anti-HIV activity and anti-HBV activity of only the 3-D-anomer of 1-dideoxy 5-fluoro-4'-thio- ⁇ -L-cytidine have been reported.
  • J. Med. Chem., (1994), 37 (6), 798-803 further describes 2 ', 3'-dideoxy L-cytidine and 2', 3'-dideoxy-l-5-fluoro-L-cytidine. It has been reported that the ⁇ -anomer and the 9-anomer have different activities in anti-HIV activity and anti-HBV activity.
  • the antiviral activity of the heavenly nucleoside cannot be referred to at all, and the synthesized ⁇ J is an a-L anomer or a ⁇ -L-anomer. Depending on this, the activity changes greatly.
  • the present inventors screened various compounds in order to find a compound having anti-hepatitis virus activity.
  • the L-1 4'-thioarabinofuranonucleoside compound showed an even higher selectivity.
  • the present inventors have found that they have hepatitis virus activity and have made the present invention. It has been reported that L-arabinofuranonucleoside compounds have no significant antiviral activity against various RNA and DNA viruses (excluding hepatitis virus) (NUCLEOSIDES & NUCLEOTIDES, 10 (6). 1345 -1376 (1991)), which was quite surprising. Disclosure of the invention
  • the present invention based on the above findings provides an L-1 4′-thioarabinofuranonucleoside-modified ⁇ I represented by the following formula [I] and a medicine containing the compound as an active ingredient! ! ⁇ Products, in particular, anti-hepatitis virus compositions ⁇
  • B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine, and azapurine, and is a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkynyl group, an amino group.
  • the L-4′-thioarabinofuranonucleoside compound of the present invention is an L-arabinofuranonucleoside compound represented by the above formula [I].
  • the base represented by B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine, and azapurine, and includes a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, and a haloalkenyl.
  • Examples of the halogen atom as a substituent include chlorine, fluorine, iodine, and bromine.
  • Examples of the alkyl group include ⁇ -alkyl groups having 1 to 7 carbon atoms, such as methyl and ethyl.
  • Examples of the haloalkyl group include a haloalkyl group having an alkyl of 1 to 1 such as fluoromethyl, difluoromethyl, trifluoromethyl, bromomethyl, and bromoethyl.
  • Examples of the alkenyl group include alkenyl groups having 2 to 7 carbon atoms, such as butyl and aryl.
  • haloalkenyl groups examples include haloalkenyl groups having a C2-7 alkenyl group such as bromovinyl and chlorovinyl.
  • alkynyl group examples include alkynyl groups having 2 to 7 ⁇ such as ethynyl and propynyl.
  • Examples of the alkylamino group on the ano group include an alkylamino group having an alkyl group having 1 to more than 1 such as methylamino and ethylamino.
  • Examples of the alkoxy group include C1 to C7 alkoxy groups such as methoxy and ethoxy.
  • Examples of the alkyl mercapto group include alkyl mercapto groups having an alkyl group having 1 to 7 carbon atoms, such as methyl mercapto and ethyl mercapto.
  • aryl group examples include a phenyl group; an alkylphenyl group having 1 to 5 carbon atoms such as methylphenyl and ethenylphenyl; an alkoxyphenyl group having 1 to 5 carbon atoms such as methoxyphenyl and ethoxyphenyl; dimethylaminophenyl and getylaminophenyl.
  • Alkylaminophenyl groups having an alkylamino having 1 to 5 carbon atoms, such as halogenophenyl groups such as chlorophenyl and promophenyl; and specific examples of pyrimidine bases include cytosine, peracyl, 5-fluorocytosine, and 5-—.
  • purine bases include purine, 6-aminopurine (adenine), 6-hydroxypurine, 6-fluoropurine, 6-chloropurine, 6-methylaminopurine, 6-dimethylaminopurine, and 6-trifluo.
  • azapurine base and the dazapurine base include 6-amino-3-dazapurine, 6-amino-8-azapurine, 2-amino-6-hydroxy-18-azapurine, 6-amino-7-dazapurine, and 6-amino-1. —Dazaprine, 6-amino-12-azapurine and the like.
  • the L-4′-thioarabinofuranonucleosides according to the present invention may be any of ⁇ -L-14′-thioarabinofuranonucleosides or ⁇ -1L-4′-thioarabinofuranonucleosides.
  • a-L—4′—thioarabinofuranopyrimidine nucleoside, a—L—4′—thioarabinofuranopurine nucleoside, —L-1 4 '-Thioarabinofuranopurine nucleoside is preferred, and particularly preferred is ⁇ -L-4'-thioarabinofuranopyrimidine nucleoside.
  • the L-4′-thioarabinofuranonucleoside compound used in the present invention may be in the form of a salt, hydrate or solvate.
  • salts include acid addition salts with inorganic acids (hydrochloric acid, sulfuric acid, phosphoric acid, etc.) or organic acids (fumaric acid, tartaric acid, conodic acid, etc.), sodium salts such as sodium salts and potassium salts.
  • alkaline earth metal salts such as lithium metal salts, calcium salts, magnesium salts and the like, and pharmaceutically acceptable salts such as ammonium salts.
  • Examples of the hydrate or solvate include those obtained by adhering 0.1 to 3. water or a solvent to the compound of the present invention or a salt thereof.
  • the L-1 4′-thioarabinofuranonucleosides of the present invention are explained below. It can be synthesized from four steps.
  • Second step (In the formula, 1 ⁇ represents an alkyl group.)
  • a step of obtaining a compound represented by the formula [III] by treating the hydroxyl group of the compound represented by the formula [II] with a mesylating agent or by subjecting the hydroxyl group to a tosyliding with a tosylating agent, followed by treating with sodium sulfate.
  • D-xylose is alkylated at the 1-position in an alcohol solvent in the presence of an acid, and then the hydroxyl group is protected. Then, the 1-alkyl group is hydrolyzed with a hidden medium and then treated with a reducing agent. This is a step of obtaining a compound represented by the formula [II].
  • Alkylation at the 1-position and protection of water can be performed according to known methods. That is, the alkylation at the 1-position is performed by converting D-xylose into an alcoholic solvent such as methanol, ethanol, or isopropanol, or an organic acid such as p-toluenesulfonic acid, drunkic acid, trifluorofluoroacid, methanesulfonic acid, or hydrochloric acid, or sulfuric acid.
  • the treatment can be carried out at a temperature of 120 ° C. to 100 ° C. in the presence of a mineral acid.
  • the protection of the hydroxyl group is carried out in a single solvent such as THF, DMF or DMSO or in a mixed solvent, in the presence of a base such as sodium hydride, or in the presence of a base such as sodium hydride or an alkyl ether such as benzyl chloride or p-methoxybenzyl chloride.
  • Hil can be formed by reacting the dangling agent at 0 to 40 ° C. with 2 to 10 mol, preferably 3 to 6 mol, per mol of D-xylose.
  • hydrochloric acid As the acid used in the hydrolysis reaction of the 1-position alkyl group, hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid and the like can be mentioned.
  • the hydrolysis reaction can be carried out by reacting the acid catalyst at 10 to 100 ° C. in a mixed solvent of a water-soluble ether solvent such as tetrahydrofuran, dioxane and the like and an aqueous acid solution.
  • a water-soluble ether solvent such as tetrahydrofuran, dioxane and the like
  • the reducing agent used in the reduction include lithium borohydride, sodium borohydride, calcium borohydride, zinc borohydride, aluminum lithium hydride, diisobutylaluminum hydride, and the like.
  • the reduction is carried out in an alcoholic solvent such as methanol or ethanol or an ethereal solvent such as ether, dioxane or THF in a reduction of 1 to 20 moles, preferably 1 to 10 moles, per mole of D-xylose.
  • Agent one hundred to one hundred, More preferably, the reaction can be carried out at a temperature of 80 ° C to 80 ° C.
  • Isolation of the thus prepared compound of formula [II] may be carried out using a conventional sugar separation / purification method. For example, after partitioning with ethyl acetate and water, the mixture is subjected to silica gel column chromatography. And elution with an organic solvent such as ⁇ -hexane monoethyl acetate can be used for separation and purification.
  • the second step is a reaction step in which the hydroxyl group of the compound of the formula [II] is mesylated or tosylated, and then treated with sodium sulfide to obtain the compound of the formula [III].
  • Mesirui-dani and tosylation can be performed based on known methods. For example, in the presence of a base such as pyridine or triethylamine in a solvent such as methylene chloride, acetonitrile, or dimethylformamide, 2 to 10 moles, preferably 2 to 4 moles, per mole of the compound of the formula [II] is used.
  • the reaction can be carried out using a molar mesylating agent (such as mesyl chloride) or a tosylating agent at 0 to 100 ° C.
  • Sulfuric acid Treatment with sodium is performed in dimethylformamide or dimethylsulfoxide in an inert gas atmosphere such as argon or nitrogen, if necessary, in an amount of 1 to 20 moles per mole of the compound of the formula [II].
  • the reaction can be carried out at room temperature to a reaction temperature of 150 using sodium sulfide.
  • the compound of the formula [III] thus prepared may be isolated by a conventional means for separating and purifying a saccharide. For example, after partitioning with ethyl acetate and water, silica gel column chromatography is performed. — Separation and purification can be achieved by elution with an organic solvent such as hexane monoethylate. Third step:
  • the compound of the formula [III] is converted to a sulfoxide form with an oxidizing agent, and further treated with an acid anhydride to perform Pummeler rearrangement to obtain a compound of the formula [IV]. is there.
  • the derivation of the compound of the formula [III] into a sulfoxide may be carried out according to a conventional method.
  • m-chloroperbenzoic acid in methylene chloride under a flow of an inert gas such as argon or nitrogen at -100 to 0 ° C. It can be carried out by treating with an acid or treating with sodium metaperiodate in an alcoholic solvent such as methanol.
  • the Pummeler rearrangement reaction may be similarly carried out according to a conventional method.
  • a compound represented by the formula [IV] can be obtained by treating in an acid anhydride such as anhydrous acetic acid at 60 ° C. to reflux.
  • the compound of the formula [IV] thus prepared may be isolated using a usual sugar separation / purification method. For example, after neutralization, the organic solvent is distilled off, and then the mixture is subjected to chromatography. It can be extracted from the aqueous layer and separated and purified by silica gel column chromatography. Step 4:
  • the fourth step is a step of introducing a nucleobase to the compound of the formula [IV] by a glycosylation reaction, and further deprotecting the sugar moiety protecting group to obtain the compound of the formula [I].
  • the glycosylation reaction is carried out under a stream of an inert gas such as argon or nitrogen, in a solvent such as methylene chloride, chloroform, dichloroethane, acetonitrile, or dimethylformamide, relative to 1 mole of the compound of the formula [IV].
  • an inert gas such as argon or nitrogen
  • a solvent such as methylene chloride, chloroform, dichloroethane, acetonitrile, or dimethylformamide, relative to 1 mole of the compound of the formula [IV].
  • a nucleic acid base which has been silylated or acylated to form a trimethylsilyl trifluoromethanesulfonate, tin tetrachloride, titanium tetrachloride, titanium chloride, kffi lead, boron trifluoride, etc.
  • isocyanic acid —50 to: can be carried out by treating at L 0 ° C.
  • Deprotection of the sugar moiety protecting group can be carried out by a conventional method depending on the protecting group used. For example, in the case of a benzyl-based protecting group, it should be treated with boron chloride or boron tribromide at ⁇ 100 ° C. to room temperature in an inert gas stream such as argon or nitrogen in methylene chloride. Can deprotect the benzyl protecting group.
  • the L-1 4'-thioarabinofuranonucleosides ⁇ ! can be separated and purified by a method suitable for the isolation and purification of nucleosides of ⁇ . After evaporating the solvent, purify by silica gel column, reverse layer column chromatography, ion exchange column chromatography, adsorption column chromatography such as active column, etc., and crystallize from an appropriate solvent such as ethanol. Depending on: ⁇ can also be obtained.
  • the dosage of the L-14'-thioarabinofuranonucleoside compound will vary depending on the patient's age and weight, disease, severity of the patient, drug tolerability, administration method, etc. Force, which is appropriately determined in the following ⁇ , usually 0.01 to 100 mg / kg body weight per day, preferably 0.01 to: L 0 mg / kg body weight It is administered once or in divided doses.
  • the method of administration can be oral, parenteral, enteral, topical or any other route.
  • Carriers include lactose, kaolin, sucrose, crystalline cellulose, corn starch, talc, agar, pectin, stearic acid, magnesium stearate, lecithin, sodium chloride
  • solid carriers such as thorium
  • liquid carriers such as glycerin, oil drop, polyvinylpyrrolidone, olive oil, ethanol, benzyl alcohol, propylene glycol, and water.
  • the dosage form can take any form.
  • a solid carrier when used, a powdery carrier, a granule, an encapsulating agent, a suppository, a troche, and the like are used.
  • examples include syrup, emulsion, soft gelatin capsule, cream, gel, paint, spray, injection and the like.
  • the solvent was distilled off, the residue was dissolved in 100 ml of DMF, 8.93 g of sodium sulfide nonahydrate was added, and the mixture was reacted at 100 ° C. for 3 hours. After the reaction, the temperature was returned to room temperature, the solvent was distilled off, the residue was dissolved in ethyl acetate, washed with water three times and saturated water, and the organic layer was dried over sodium sulfate. After evaporating the solvent, the residue was purified by a silica gel column to obtain the title compound (4.38 g, 84%).
  • the reaction was stopped by adding a saturated sodium hydrogen carbonate solution, and the insolubles were filtered through celite. The filtrate was extracted three times with a black hole form, and dried. After concentration, purification was carried out using a silica gel column to obtain 445 mg (78%) of a nucleoside conjugate.
  • the nucleoside compound (417 mg) was dissolved in methylene chloride (1 Oml) and cooled to 178. To this solution, 4.38 ml of a 1M methylene chloride solution was added dropwise and stirred at 178 ° C for 30 minutes. The temperature was raised to 120 ° C, and the mixture was further stirred for 3 hours, and the reaction was stopped by adding a saturated sodium hydrogen carbonate solution. The solvent was distilled off, 5 ml of methanol and 5 ml of concentrated aqueous ammonia were added to the residue, and the mixture was stirred at room temperature for 10 minutes. The solvent was distilled off, and the residue was azeotroped with ethanol three times, and purified by a silica gel column. The obtained anomeric mixture was purified by ODS column chromatography and the anomers were separated to obtain the enantiomer of the title ⁇ ! (64 mg, 34%) and the) S-anomer (36 mg, 19%).
  • Adenine (16 mg) and ⁇ anomer (106 mg, crude) as the title compounds were obtained in the same manner as described above, except that adenine was used instead of N 4 -acetylcytosine.
  • HB611 Proc. Natl. Acad. Sci. USA 84, 444-448 (1987)
  • the method of Shuto et al. (Microbiol. Immunol., 40 (2), 153-159 (1996)), and the anti-V activity of the drug was measured.
  • ⁇ 611 cells were suspended in Dulbecco's modified MEM containing 10% bovine i3 ⁇ 4fil, 1001 UZm1 benisiri G, 100 g / m1 streptomycin and lmg / m1 geneticin. 20,000 pieces per 1 well of a 96-well multiwell plate were applied.
  • the cells were cultured in a carbon dioxide incubator at 37 ° C. for 3 days, and the cells were grown to confluent and then subjected to a test.
  • the test drug was added to the cells, usually diluted with medium, in four steps of 10-fold serial dilution from a concentration of 100 / ig / inl, and cultured. The medium was replaced two to four days later with the medium containing the drug. Control cells were cultured in a drug-free medium. Seven days later, the culture supernatant was recovered from each well and transferred to another 96-well multiwell plate. Saved in C.
  • Anti-HBV surface antigen mouse antibody diluted to 10 ( ⁇ gZml) with isotonic phosphate buffer (PBS) in each well of a 96-well multiwell plate (Korning)
  • PCR For PCR, a commercially available kit, TaKaRa Taq (manufactured by Takara Shuzo) and Gene Amp system 9600 (manufactured by Perkin Elmer) were used.
  • the primers used were designed so that Nos. 372 to 483 in the S residue of HBV were amplified. That is, the primer of (+) 372-401 (5'-biotin-TCGCTGGATGTGTCTGCGGCGTTTT AT) and the primer (1) of 460-483 (5'-TAGAGGACAA Using.
  • PCR mixed solution 45 ⁇ 1 (Fiber: 10 mM Tris-monohydrochloride buffer (pH 8.3), 5 OmM KC 1, 1.5 mM MgCl 2 , 0.2 mM of DNA dNTP, 6.25 pmo 1 of each primer, 1.25 U Amp 1 i Taq DNA polymerase) and PCR. 94. C. After 5 minutes incubation, 30 cycles of 94, 30 seconds and 55, 15 seconds—72 ° C., 1 minute were performed, and finally the reaction was performed at 72 ° C. for 5 minutes, and stored at 4.
  • Streptavidin diluted to lO ⁇ gZml with 5 OmM quenched buffer (pH 9.6) in each well of a flat-bottomed 96-well multiwell plate (manufactured by Koingen) was added at 75 ° C. each, and the mixture was left at 4 ° C. for about 16 hours. The liquid layer in each well was discarded, PBS 100 // 1 containing 0.1% BSA was added, the mixture was incubated at 37 ° C for 2 hours, and stored at 4 ° C until immediately before use. After washing twice with 0.1 XS SPE solution containing 0.1% Tween 20 (0.1 XS SPE-0.
  • 0.1 XS SPE After washing 3 times with 0.05% Tween 20 and 2 times with PBS, add 100 1 of 2000-fold diluted alkaline phosphatase-conjugated anti-dioxygenin antibody (manufactured by Boehringer Mannheim GmbH) and add room temperature. For 1 hour. After washing 4 times with 0. 1 XS S PE- 0. 05% Twe en 20, 1 0 OmM Na C 1 and 5 OmM Mg C 1 2 and 100 mM Tris monohydrochloride buffer containing of (pH 9. 5 ), Adjusted to 1 mg / m 1 with 1002 trophenylphosphite solution 1001, and incubated at 37 ° C for 5 to 15 minutes until color development.
  • the absorbance at 405 nm of each well was measured with a microplate reader to calculate the anti-HBV activity of the test drug.
  • the concentration of the test drug that reduces the absorption of the control to which no drug was added by 50% was increased by the 50% effective concentration (EC 50)
  • the cytotoxicity of the test drug was measured by the MTT method. After the culture supernatant was recovered, HB61 1 cells were added with a fresh medium 100 1 and a 7.5 mg / m 1 MTT solution 201, and cultured at 37 ° C for 3 hours in a carbon dioxide gas incubator. Formalzan was eluted by adding 10 1 of a hidden isopropanol (500 ml of isopropanol to which 2 ml of concentrated hydrochloric acid was added) containing 10% (v / v) Triton X-100. After the formasan was completely dissolved, the absorbance at 540 and 690 nm was measured using a microplate reader to calculate the cytotoxicity of the test drug. The test drug that reduced the absorbance of the control to which no drug was added by 50% was defined as 50% cytotoxicity i «(cc 50 ).
  • Fine powdered cellulose 25.Omg
  • the L-4′-thioarabinofuranonucleoside compound of the present invention has no toxicity and has a remarkable inhibitory effect on hepatitis B virus (HBV).
  • Hepatitis virus ⁇ especially anti-hepatitis B virus It is useful as an agent and can be expected to be developed as a pharmaceutical.
  • the method for synthesizing the above-mentioned L-4′-thioarabinofuranonucleoside compound is an extremely practical method that can derive the desired compound from the natural sugar D-xylose by a simple method.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

A L-4'-thioarabinofuranonucleoside compound represented by formula (1) wherein B represents a nucleic acid base selected from among pyrimidine, purine, azapurine and deazapurine, each of which may be substituted with a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkinyl group, an amino group, an alkylamino group, a hydroxyl group, a hydroxyamino group, an aminoxy group, an alkoxy group, a mercapto group, an alkylmercapto group, an aryl group, an aryloxy group or a cyano group, and a medicine composition comprising the compound as an active component, especially antihepatitis virus composition.

Description

明 細 書  Specification
L— 4 ' ーチオアラビノフラノヌクレオシドィ匕合物および それを含む医薬組成物 饮 分 野 L-4′-thioarabinofuranonucleoside conjugates and pharmaceutical compositions containing the same
本発明は、 L— 4 ' ーチオアラビノフラノヌクレオシド化合物および該化合物 を活性 として含有する医薬組成物、 特に、 抗肝炎ウィルス剤に関するもので あ 背 景 技 術  TECHNICAL FIELD The present invention relates to an L-4′-thioarabinofuranonucleoside compound and a pharmaceutical composition containing the compound as an activity, particularly to an anti-hepatitis virus agent.
非天然型構造を有する L -ヌクレオシドの一部に抗ゥィルス効果が認め られ、 その低い毒性と合わせて、 注目を集めている。 たとえば、 2 ' 一フルォロ —5—メチル一 3— L—ァラビノフラノシルゥラシル （L— F MA U) は抗 B型 ウィルス （H B V) 活性を有し、 しかも細胞毒性が低いことが報告されてい る （Antimicrob. Agents Chemother. , (1995), 39 (4) , 979-981： Antimicrob. Agents Chemother. , (1996) , 40(2) , 380-386 )。  Some L-nucleosides having an unnatural structure have been shown to have an anti-viral effect, and are attracting attention due to their low toxicity. For example, it has been reported that 2'-fluoro-5-methyl-13-L-arabinofuranosylperacil (L-FMAU) has anti-B virus (HBV) activity and low cytotoxicity. Agents Chemother., (1995), 39 (4), 979-981: Antimicrob. Agents Chemother., (1996), 40 (2), 380-386.
しかしながら、 非天然 構造の Lーヌクレオシドの場合にはターゲッ トウィル ス及びその活性を予想することは天然型の化合物以上に困難である。  However, in the case of L-nucleosides having a non-natural structure, it is more difficult to predict the target virus and its activity than the natural compound.
たとえば、 Antiviral Chemistry & Chemotherapy (1991)， 2(2), 83-92 には、 A Z Tの誘導体として抗 H I V活性を期待して合成した 3 ' —置換チミジンの α - L—ヌクレオシド誘導体が、 顕著な抗ウィルス活性を示さないことが報告され ている。  For example, in Antiviral Chemistry & Chemotherapy (1991), 2 (2), 83-92, α-L-nucleoside derivatives of 3'-substituted thymidine synthesized with the expectation of anti-HIV activity as a derivative of AZT are remarkable. It has been reported that it does not show antiviral activity.
また、 Antiviral Chemistry & Chemotherapy (1995) , 6(3)， 138-142 には、 a 一 L—ァラビノフラノシルシトシン、 α— L—キシロフラノシルシトシンおよび a— L— FMAUの 一 L—ァノマーに若干の抗 HCMV (ヒトサイトメガロウ ィルス） 活性が確認されたものの、 上記以外の α— Lーァノマーおよび上記化合 物の ^— Lーァノマーには抗 H CMV活性は確認されていない。 In addition, Antiviral Chemistry & Chemotherapy (1995), 6 (3), 138-142 include a-L-arabinofuranosylcytosine, α-L-xylofuranosylcytosine and Although some anti-HCMV (human cytomegalovirus) activity was confirmed in one L-anomer of a-L-FMAU, anti-H CMV was observed in other α-L anomers and ^ -L anomers of the above compounds. No activity has been confirmed.
さらに、 特表平 8— 504753号公報では、 2' , 3' 一ジデヒドロ— 2' ， 3' —ジデォキシ一 4' 一チオー^一 L—シチジンおよび 2' ， 3' ージデヒド ロー 2' , 3' 一ジデォキシー 5—フルオロー 4' 一チオー^— L—シチジンの ;3— L—ァノマーのみ抗 H I V活性および抗 HBV活性が報告されている。  Furthermore, Japanese Patent Publication No. 8-504753 discloses that 2 ', 3'-didehydro-2', 3'-dideoxy-14'-thio-^-l-cytidine and 2 ', 3'didehydro 2', 3 ' The anti-HIV activity and anti-HBV activity of only the 3-D-anomer of 1-dideoxy 5-fluoro-4'-thio-^-L-cytidine have been reported.
さらにまた、 J. Med. Chem. , (1994), 37(6) , 798-803 には、 2' , 3' — ジデォキシー Lーシチジンおよび 2' , 3' —ジデォキシ一 5—フルオロー L— シチジンの抗 H I V活性および抗 HBV活性において、 α—ァノマーと) 9ーァノ マーでそれらの活性が異なることが報告されている。  Furthermore, J. Med. Chem., (1994), 37 (6), 798-803 further describes 2 ', 3'-dideoxy L-cytidine and 2', 3'-dideoxy-l-5-fluoro-L-cytidine. It has been reported that the α-anomer and the 9-anomer have different activities in anti-HIV activity and anti-HBV activity.
このように非天 ^[構造の L—ヌクレオシドの場合、 天 のヌクレオシドの 抗ウィルス活性は何等参考にすることができず、 また、 合成した化^ Jが a— L ーァノマーか^— L—ァノマーかによつてもその活性は大きく変化する。  Thus, in the case of an L-nucleoside having a non-heavenly ^ [structure, the antiviral activity of the heavenly nucleoside cannot be referred to at all, and the synthesized ^ J is an a-L anomer or a ^ -L-anomer. Depending on this, the activity changes greatly.
このような状況下で、 本発明者らは、 抗肝炎ウィルス活性を有する化合物を見 出すべく、 種々の化合物をスクリーニングした結果、 L一 4' —チオアラビノフ ラノヌクレオシド化合物が選択性の一段と高い抗 Β型肝炎ゥィルス活性を有する ことを見出し、 本発明を^させた。 L—ァラビノフラノヌクレオシド化合物に 関しては種々の RNAおよび DNAウィルス （肝炎ウィルスを除く） に対して有 意な抗ウィルス活性を示さないことが報告 (NUCLEOSIDES & NUCLEOTIDES, 10(6) . 1345-1376(1991))されていただけに、 全く意外な結果であつた。 発 明 の 開示  Under such circumstances, the present inventors screened various compounds in order to find a compound having anti-hepatitis virus activity. As a result, the L-1 4'-thioarabinofuranonucleoside compound showed an even higher selectivity. The present inventors have found that they have hepatitis virus activity and have made the present invention. It has been reported that L-arabinofuranonucleoside compounds have no significant antiviral activity against various RNA and DNA viruses (excluding hepatitis virus) (NUCLEOSIDES & NUCLEOTIDES, 10 (6). 1345 -1376 (1991)), which was quite surprising. Disclosure of the invention
上記の知見に基づく本発明は、 下記式 [I] で表される L一 4' ーチオアラビ ノフラノヌクレオシド化^ Iおよび該化合物を活1«分として含有する医！!^ 物、 特に、 抗肝炎ウィルス組成物を提供するものである < The present invention based on the above findings provides an L-1 4′-thioarabinofuranonucleoside-modified ^ I represented by the following formula [I] and a medicine containing the compound as an active ingredient! ! ^ Products, in particular, anti-hepatitis virus compositions <
Figure imgf000005_0001
Figure imgf000005_0001
(式中、 Bはピリミジン、 プリン、 ァザプリンおよびデァザプリンからなる群よ り選択される核酸塩基を示し、 それらはハロゲン原子、 アルキル基、 ハロアルキ ル基、 アルケニル基、 ハロアルケニル基、 アルキニル基、 アミノ基、 アルキルァ ミノ基、 水酸基、 ヒドロキシァミノ基、 アミノキシ基、 アルコキシ基、 メルカプ 卜基、 アルキルメルカプト基、 ァリール基、 ァリールォキシ基またはシァノ基に よって置換されていてもよい。 ） 発明を実施するための最良の形態 (Wherein, B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine, and azapurine, and is a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkynyl group, an amino group. , An alkylamino group, a hydroxyl group, a hydroxyamino group, an aminoxy group, an alkoxy group, a mercapto group, an alkylmercapto group, an aryl group, an aryloxy group or a cyano group.) Best form
( 1 ) L - 4 ' —チオアラビノフラノヌクレオシド化合物  (1) L-4'-thioarabinofuranonucleoside compound
本発明の L— 4 ' —チオアラビノフラノヌクレオシド化合物は上記式 [ I ] で 表される L—ァラビノフラノヌクレオシド化合物である。  The L-4′-thioarabinofuranonucleoside compound of the present invention is an L-arabinofuranonucleoside compound represented by the above formula [I].
式中、 Bで表される塩基としてはピリ ミジン、 プリン、 ァザプリンおよびデァ ザプリンからなる群より選択される核酸塩基を示し、 それらはハロゲン原子、 ァ ルキル基、 ハロアルキル基、 アルケニル基、 ハロアルケニル基、 アルキニル基、 アミノ基、 アルキルアミノ基、 水酸基、 ヒドロキシァミノ基、 アミノキシ基、 ァ ルコキシ基、 メルカプト基、 アルキルメルカプト基、 ァリール基、 ァリールォキ シ基またはシァノ基などの置換基を有していても構わない。 また、 そのような置 換基の位置および数は特に制限されるものではない。 In the formula, the base represented by B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine, and azapurine, and includes a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, and a haloalkenyl. Group, alkynyl group, amino group, alkylamino group, hydroxyl group, hydroxyamino group, aminoxy group, alkoxy group, mercapto group, alkylmercapto group, aryl group, aryloxy group It may have a substituent such as a cyano group or a cyano group. Further, the position and number of such a substituent are not particularly limited.
置換基としてのハロゲン原子としては、 塩素、 フッ素、 ヨウ素、 臭素が例示さ れる。 アルキル基としては、 メチル、 ェチルなどの炭素数 1〜 7の βアルキル 基が例示される。 ハロアルキル基としては、 フルォロメチル、 ジフルォロメチル、 トリフルォロメチル、 プロモメチル、 プロモェチルなどの^^数 1〜了のアルキ ルを有するハロアルキル基力例示される。 アルケニル基としては、 ビュル、 ァリ ルなどの炭素数 2〜 7のァルケニゾレ基が例示される。 ハロアルケニル基としては、 ブロモビニル、 クロ口ビニルなどの炭素数 2〜 7のァルケ二ル基を有するハ口了 ルケニル基が例示される。 アルキニル基としては、 ェチニル、 プロピニルなどの ^数 2〜 7のアルキニル基が例示される。  Examples of the halogen atom as a substituent include chlorine, fluorine, iodine, and bromine. Examples of the alkyl group include β-alkyl groups having 1 to 7 carbon atoms, such as methyl and ethyl. Examples of the haloalkyl group include a haloalkyl group having an alkyl of 1 to 1 such as fluoromethyl, difluoromethyl, trifluoromethyl, bromomethyl, and bromoethyl. Examples of the alkenyl group include alkenyl groups having 2 to 7 carbon atoms, such as butyl and aryl. Examples of the haloalkenyl groups include haloalkenyl groups having a C2-7 alkenyl group such as bromovinyl and chlorovinyl. Examples of the alkynyl group include alkynyl groups having 2 to 7 ^ such as ethynyl and propynyl.
ァノ上キルアミノ基としては、 メチルァミノ、 ェチルァミノなどの^数 1〜了 のアルキル基を有するアルキルァミノ基が例示される。 アルコキシ基としては、 メ トキシ、 エトキシなどの炭素数 1〜 7のアルコキシ基が例示される。 アルキル メルカプト基としては、 メチルメルカプト、 ェチルメルカプトなどの炭素数 1〜 7のアルキル基を有するアルキルメルカプト基が例示される。 ァリール基として は、 フエニル基；メチルフヱニル、 ェチルフエニルなどの炭素数 1〜5のアルキ ルを有するアルキルフヱニル基； メ トキシフヱニル、 エトキシフヱニルなどの炭 素数 1〜5のアルコキシフエニル基； ジメチルァミノフエニル、 ジェチルァミノ フエニルなどの炭素数 1〜 5のアルキルアミノを有するアルキルァミノフエニル 基； クロロフヱニル、 プロモフヱニルなどのハロゲノフエニル基などが例示され ピリミジン塩基を具体的に例示すれば、 シトシン、 ゥラシル、 5—フルォロシ トシン、 5—フルォロウラシル、 5—クロロシトシン、 5—クロロウラシル、 5 —プロモシトシン、 5—プロモウラシル、 5—ョードシトシン、 5—ョードウラ シル、 5—メチルシトシン、 5—メチルゥラシル （チミ ン） 、 5—ェチルシトシ ン、 5—ェチルゥラシル、 5—フルォロメチルシトシン、 5—フルォロメチルゥ ラシル、 5—トリフルォロシトシン、 5—トリフルォロウラシル、 5—ビニルゥ ラシル、 5—プロモビニルゥラシル、 5—クロロビニルゥラシル、 5—ェチニル シトシン、 5—ェチニルゥラシル、 5—プロピニルゥラシル、 ピリ ミジン一 2— オン、 4ーヒ ドロキシァミノピリ ミジン一 2—オン、 4—アミノォキシピリ ミジ ン一 2—オン、 4—メ トキシピリ ミジン一 2—オン、 4—ァセトキシピリ ミジン —2—オン、 5—フルォロピリ ミジン一 2—オンなどが挙げられる。 Examples of the alkylamino group on the ano group include an alkylamino group having an alkyl group having 1 to more than 1 such as methylamino and ethylamino. Examples of the alkoxy group include C1 to C7 alkoxy groups such as methoxy and ethoxy. Examples of the alkyl mercapto group include alkyl mercapto groups having an alkyl group having 1 to 7 carbon atoms, such as methyl mercapto and ethyl mercapto. Examples of the aryl group include a phenyl group; an alkylphenyl group having 1 to 5 carbon atoms such as methylphenyl and ethenylphenyl; an alkoxyphenyl group having 1 to 5 carbon atoms such as methoxyphenyl and ethoxyphenyl; dimethylaminophenyl and getylaminophenyl. Alkylaminophenyl groups having an alkylamino having 1 to 5 carbon atoms, such as halogenophenyl groups such as chlorophenyl and promophenyl; and specific examples of pyrimidine bases include cytosine, peracyl, 5-fluorocytosine, and 5-—. Fluorouracil, 5-chlorocytosine, 5-chlorouracil, 5—promocytosine, 5-promouracil, 5-ododocytosine, 5-ododoura Syl, 5-methylcytosine, 5-methylperacyl (thymine), 5-ethylethylcytosine, 5-ethylperacyl, 5-fluoromethylcytosine, 5-fluoromethylperacil, 5-trifluoromethylcytosine, 5-trifluorouracil , 5-vinyl-racil, 5-promovinyl-racil, 5-chlorovinyl-peracyl, 5-ethynylcytosine, 5-ethynyl-peracyl, 5-propynyl-peracyl, pyrimidine-2-one, 4-hydroxyhydroxyaminopyrimidine Examples include 12-one, 4-aminooxypyrimidine-12-one, 4-methoxypyrimidine-12-one, 4-acetoxypyrimidine-2-one, and 5-fluoropyrimidin-12-one.
プリン塩基を具体的に例示すれば、 プリン、 6—ァミノプリン （アデニン） 、 6—ヒ ドロキシプリン、 6—フルォロプリン、 6—クロ口プリン、 6—メチルァ ミノプリン、 6—ジメチルァミノプリン、 6—トリフルォロメチルァミノプリン、 6—べンゾィルァミノプリン、 6—ァセチルァミノプリン、 6—ヒ ドロキシアミ ノプリン、 6—アミノォキシプリン、 6—メ トキシプリン、 6—ァセトキシプリ ン、 6—ベンゾィルォキシプリン、 6—メチルプリン、 6—ェチルプリン、 6— トリフルォロメチルプリン、 6—フエ二ルブリン、 6—メルカプトプリン、 6— メチルメルカプトプリン、 6—ァミノプリン一 1ーォキシド、 6—ヒドロキシプ リン一 1ーォキシド、 2—アミノー 6—ヒドロキシプリン （グァニン） 、 2—ァ ミノプリン、 2， 6—ジァミノプリン、 2—アミノー 6—クロ口プリン、 2—ァ ミノ一 6—ョ一ドブリン、 2—ァミノプリン、 2—アミノー 6—メルカプトプリ ン、 2—ァミノ一 6—メチルメルカプトプリン、 2—アミノー 6—ヒドロキシァ ミノプリン、 2—アミノー 6—メ 卜キシプリン、 2—アミノー 6—ベンゾィルォ キシプリン、 2—アミノー 6—ァセトキシプリン、 2—アミノー 6—メチルプリ ン、 2—ァミノ一 6—サイクロプロピルアミノメチルブリン、 2—アミノー 6— フエ二ルブリン、 2—アミノー 8—プロモプリン、 6—シァノプリン、 6—アミ ノー 2—クロ口プリン、 6—ァミノ一 2—フルォロプリンなどが挙げられる。 ァザプリン塩基およびデァザプリン塩基を具体的に例示すれば、 6—アミノー 3—デァザプリン、 6—アミノー 8—ァザプリン、 2—ァミノ一 6—ヒドロキシ 一 8—ァザプリン、 6—アミノー 7—デァザプリン、 6—アミノー 1—デァザプ リン、 6—ァミノ一 2—ァザプリンなどが挙げられる。 Specific examples of purine bases include purine, 6-aminopurine (adenine), 6-hydroxypurine, 6-fluoropurine, 6-chloropurine, 6-methylaminopurine, 6-dimethylaminopurine, and 6-trifluo. Romethylaminopurine, 6-Benzoylaminopurine, 6-Acetylaminopurine, 6-Hydroxyaminopurine, 6-Aminoxypurine, 6-Methoxypurine, 6-Acetoxypurine, 6-Benzo Yloxypurine, 6-methylpurine, 6-ethylpurine, 6-trifluoromethylpurine, 6-phenylurine, 6-mercaptopurine, 6-methylmercaptopurine, 6-aminopurine 1-oxide, 6-hydroxyp Phosphorus 1-oxide, 2-amino-6-hydroxypurine (guanine), 2-aminopurine, 2,6-diamino Purine, 2-amino-6-clonal purine, 2-amino-6-phosphoridoline, 2-aminopurine, 2-amino-6-mercaptopurine, 2-amino-6-methylmercaptopurine, 2-amino-6- Hydroxyaminopurine, 2-amino-6-methoxypurine, 2-amino-6-benzoyloxypurine, 2-amino-6-acetoxypurine, 2-amino-6-methylpurine, 2-amino-6-cyclopropylaminomethylbrin, 2- Examples include amino-6-phenylurine, 2-amino-8-promopurine, 6-cyanopurine, 6-amino-2-chloropurine, and 6-amino-12-fluoropurine. Specific examples of the azapurine base and the dazapurine base include 6-amino-3-dazapurine, 6-amino-8-azapurine, 2-amino-6-hydroxy-18-azapurine, 6-amino-7-dazapurine, and 6-amino-1. —Dazaprine, 6-amino-12-azapurine and the like.
本発明の L— 4 ' —チオアラビノフラノヌクレオシドィ匕^!としては、 α— L 一 4 ' ーチオアラビノフラノヌクレオシドあるいは ^一 L— 4' ーチオアラビノ フラノヌクレオシドのいずれであってもよい。 なお、 前言己式 （I ) 中の波線 The L-4′-thioarabinofuranonucleosides according to the present invention may be any of α-L-14′-thioarabinofuranonucleosides or ^ -1L-4′-thioarabinofuranonucleosides. The wavy line in the self-expression (I)
( ) は、 α—ァノマ一或いは^—ァノマーのいずれであってもよいことを意 味している。 () Means that it may be either α-anomer or ^ -anomer.
そのような化合物を活 ¾β¾分として本発明の組成物に用いる場合は、 a - L— 4' —チオアラビノフラノピリミジンヌクレオシド、 a— L— 4 ' —チオアラビ ノフラノプリンヌクレオシド、 ー L一 4' ーチオアラビノフラノプリンヌクレ オシドが好ましく、 特に好適なものとして α— L— 4 ' —チオアラビノフラノピ リミジンヌクレオシドを挙げることができる。  When such a compound is used as an active ingredient in the composition of the present invention, a-L—4′—thioarabinofuranopyrimidine nucleoside, a—L—4′—thioarabinofuranopurine nucleoside, —L-1 4 '-Thioarabinofuranopurine nucleoside is preferred, and particularly preferred is α-L-4'-thioarabinofuranopyrimidine nucleoside.
本発明で用いる L— 4 ' —チオアラビノフラノヌクレオシド化合物は、 塩、 水 和物または溶媒和物の形態であってもよい。 そのような塩としては、 無機酸 （塩 酸、 硫酸、 リン酸など） または有機酸 （フマル酸、 酒石酸、 コノヽク酸など） との 酸付加塩、 ナトリゥム塩、 力リゥム塩などのアル力リ金属塩、 カルシウム塩、 マ グネシゥム塩などのアル力リ土類金属塩、 またはアンモニゥム塩など薬学的に許 容される塩を例示することができる。  The L-4′-thioarabinofuranonucleoside compound used in the present invention may be in the form of a salt, hydrate or solvate. Examples of such salts include acid addition salts with inorganic acids (hydrochloric acid, sulfuric acid, phosphoric acid, etc.) or organic acids (fumaric acid, tartaric acid, conodic acid, etc.), sodium salts such as sodium salts and potassium salts. Examples thereof include alkaline earth metal salts such as lithium metal salts, calcium salts, magnesium salts and the like, and pharmaceutically acceptable salts such as ammonium salts.
また、 水和物または溶媒和物としては、 本発明化合物またはその塩 に対 し、 0. 1〜3. の水または溶媒が付着したものを例示することができる。  Examples of the hydrate or solvate include those obtained by adhering 0.1 to 3. water or a solvent to the compound of the present invention or a salt thereof.
( 2 ) L一 4 ' ーチオアラビノフラノヌクレオシド化合物の合成 (2) Synthesis of L-1 4'-thioarabinofuranonucleoside compound
本発明の L一 4 ' —チオアラビノフラノヌクレオシド化^!は、 以下に説明す る 4つの工程より合成することができる。 The L-1 4′-thioarabinofuranonucleosides of the present invention are explained below. It can be synthesized from four steps.
第 1工程： First step:
D—キシロースを、 アルコール溶媒中、 酸の存在下で 1位をアルキル化した後、 水酸基を保護し、 次いで 1位アルキル基の酸 による加水分解、 引き続き還元 剤による処理により式 [I I] で表される化合物を得る工程。  After alkylating D-xylose at the 1-position in an alcohol solvent in the presence of an acid, protecting the hydroxyl group, then hydrolyzing the alkyl at the 1-position with an acid, and subsequently treating with a reducing agent, the formula [II] is used. Obtaining the compound to be prepared.
D一 xyloseD-I xylose
Figure imgf000009_0001
Figure imgf000009_0001
(式中、 1^ はアルキル基を示す。 ） 第 2工程： (In the formula, 1 ^ represents an alkyl group.) Second step:
式 [I I] で表される化合物の水酸基をメシル化剤によりメシル化またはトシ ル化剤により トシルイ匕した後、 硫ィ匕ナトリウムで処理して式 [I I I] で表され る化合物を得る工程。  A step of obtaining a compound represented by the formula [III] by treating the hydroxyl group of the compound represented by the formula [II] with a mesylating agent or by subjecting the hydroxyl group to a tosyliding with a tosylating agent, followed by treating with sodium sulfate.
Figure imgf000009_0002
Figure imgf000009_0002
(式中、 Rx は前記と同義。 ) 第 3工程： (In the formula, R x is as defined above.) Third step:
式 [I I I] で表される化合物を適当な酸化剤によりスルホキシド体に導き- 更に酸無水物で処理することでプンメラー （Pumme r e r)転移を行ない- 式 [IV] で表される化合物を得る工程。  Step of leading a compound represented by the formula [III] to a sulfoxide form with an appropriate oxidizing agent-further treating the compound with an acid anhydride to carry out Pummeler rearrangement-obtaining a compound represented by the formula [IV] .
Figure imgf000010_0001
Figure imgf000010_0001
(式中、 R1 は、 前記と同義。 R2 はァシル基を示す。 ） 第 4工程： (Wherein, R 1 has the same meaning as described above. R 2 represents an acyl group.) Fourth step:
式 [IV] で表される化合物にグリコシルイ 応により核酸塩基を導入し、 更 に糖部保護基を脱保護して式 [I] で表される化合物を得る工程。  A step of introducing a nucleobase to the compound represented by the formula [IV] by glycosylation, and further deprotecting the sugar protecting group to obtain a compound represented by the formula [I].
Figure imgf000010_0002
Figure imgf000010_0002
(式中、 Bは前記と同義。 ) 以下、 第 1〜第 4工程をさらに詳しく説明する。 (In the formula, B is as defined above.) Hereinafter, the first to fourth steps will be described in more detail.
第 1工程：  First step:
第 1工程は、 D—キシロースを、 アルコール溶媒中、 酸の存在下で 1位をアル キルイ匕した後、 水酸基を保護し、 次いで 1位アルキル基の隱媒による加水分 引き続き還元剤による処理により式 [ I I ] で表される化合物を得る工程である。  In the first step, D-xylose is alkylated at the 1-position in an alcohol solvent in the presence of an acid, and then the hydroxyl group is protected. Then, the 1-alkyl group is hydrolyzed with a hidden medium and then treated with a reducing agent. This is a step of obtaining a compound represented by the formula [II].
1位のアルキル化と水^の保護については、 公知の方法に従って行うことが できる。 即ち、 1位のアルキル化は、 D—キシロースを、 メタノール、 エタノー ル、 イソプロパノールなどのアルコール溶媒中、 p—トルエンスルホン酸、 醉酸、 トリフルォロ醉酸、 メタンスルホン酸などの有機酸もしくは塩酸、 硫酸などの鉱 酸の存在下一 2 0 °C〜1 0 0 °Cで処理することにより行なうことができる。 また、 水酸基の保護は、 T H F、 DMF、 DM S Oなどの単独溶媒中もしくは混合溶媒 中、 水素化ナトリゥムなどの塩基の存在下、 ベンジルクロリ ド、 ベンジルブロミ ド、 p—メ トキシベンジルクロリ ドなどのアルキルイ匕剤を、 D—キシロース 1モ ルに対して 2〜1 0モル、 好ましくは 3〜6モル用い、 0〜4 0 °Cで反応させる ことにより Hilすることができる。  Alkylation at the 1-position and protection of water can be performed according to known methods. That is, the alkylation at the 1-position is performed by converting D-xylose into an alcoholic solvent such as methanol, ethanol, or isopropanol, or an organic acid such as p-toluenesulfonic acid, drunkic acid, trifluorofluoroacid, methanesulfonic acid, or hydrochloric acid, or sulfuric acid. The treatment can be carried out at a temperature of 120 ° C. to 100 ° C. in the presence of a mineral acid. The protection of the hydroxyl group is carried out in a single solvent such as THF, DMF or DMSO or in a mixed solvent, in the presence of a base such as sodium hydride, or in the presence of a base such as sodium hydride or an alkyl ether such as benzyl chloride or p-methoxybenzyl chloride. Hil can be formed by reacting the dangling agent at 0 to 40 ° C. with 2 to 10 mol, preferably 3 to 6 mol, per mol of D-xylose.
1位アルキル基の加水分解反応において用いる酸醒としては、 塩酸、 硫酸、 酢酸、 トリフルォロ酢酸などを挙げることができる。 加水分解反応は、 テトラヒ ドロフラン、 ジォキサンなどの水溶性エーテル系の溶媒と酸水溶液との混合溶媒 中、 1 0〜1 0 0 °Cで上記酸触媒を作用させることにより することができる。 還 ^応において使用する還元剤としては、 水素化ホウ素リチウム、 水素化ホ ゥ素ナトリウム、 水素化ホウ素カルシウム、 水素ィ匕ホウ素亜鉛、 水素化アルミ二 ゥムリチウム、 水素化ジイソプチルアルミニウムなどを挙げることができる。 還 ^応は、 メタノール、 エタノールなどのアルコール系溶媒、 もしくはエーテル、 ジォキサン、 T H Fなどのエーテル系溶媒中、 D—キシロース 1モルに対して 1 〜2 0モル、 好ましくは 1〜1 0モルの還元剤を用い、 一 1 0 0〜1 0 0 、 好 ましくは一 80°C〜80°Cで反応させることにより実施することができる。 As the acid used in the hydrolysis reaction of the 1-position alkyl group, hydrochloric acid, sulfuric acid, acetic acid, trifluoroacetic acid and the like can be mentioned. The hydrolysis reaction can be carried out by reacting the acid catalyst at 10 to 100 ° C. in a mixed solvent of a water-soluble ether solvent such as tetrahydrofuran, dioxane and the like and an aqueous acid solution. Examples of the reducing agent used in the reduction include lithium borohydride, sodium borohydride, calcium borohydride, zinc borohydride, aluminum lithium hydride, diisobutylaluminum hydride, and the like. Can be. The reduction is carried out in an alcoholic solvent such as methanol or ethanol or an ethereal solvent such as ether, dioxane or THF in a reduction of 1 to 20 moles, preferably 1 to 10 moles, per mole of D-xylose. Agent, one hundred to one hundred, More preferably, the reaction can be carried out at a temperature of 80 ° C to 80 ° C.
このようにして調製した式 [I I] のィ匕合物の単離は、 通常の糖の分離精製手 段を用いて行なえばよく、 たとえば齚酸ェチルと水で分配後、 シリカゲルカラム クロマトグラフィ一に付し、 η-へキサン一酢酸ェチルなどの有機溶媒で溶出す ることにより分離精製できる。 第 2工程：  Isolation of the thus prepared compound of formula [II] may be carried out using a conventional sugar separation / purification method. For example, after partitioning with ethyl acetate and water, the mixture is subjected to silica gel column chromatography. And elution with an organic solvent such as η-hexane monoethyl acetate can be used for separation and purification. Second step:
第 2工程は式 [I I] の化合物の水酸基をメシル化あるいはトシルイ匕した後、 硫化ナトリウムと処理して式 [I I I] で表される化合物を得る反応工程である。 メシルイ匕およびトシル化は公知の方法に基づき行なうことができる。 たとえば、 ピリジン、 トリェチルァミンなどの塩基の存在下、 塩ィ匕メチレン、 ァセトニトリ ル、 ジメチルホルムアミ ドなどの溶媒中、 式 [I I] の化合物 1モルに対して 2 〜 10モル、 好ましくは 2〜 4モルのメシル化剤 （塩化メシルなど） もしくはト シル化剤を用い、 0〜100°Cで反応させることにより実施することができる。 硫^:ナトリウムとの処理は、 ジメチルホルムアミ ドあるいはジメチルスルホキ シド中、 必要に応じてアルゴン、 窒素などの不活性ガス雰囲気下、 式 [I I] の 化合物 1モルに対して 1〜20モルの硫化ナトリウムを用い、 室温〜 150 の 反応温度で反応させることにより実施できる。  The second step is a reaction step in which the hydroxyl group of the compound of the formula [II] is mesylated or tosylated, and then treated with sodium sulfide to obtain the compound of the formula [III]. Mesirui-dani and tosylation can be performed based on known methods. For example, in the presence of a base such as pyridine or triethylamine in a solvent such as methylene chloride, acetonitrile, or dimethylformamide, 2 to 10 moles, preferably 2 to 4 moles, per mole of the compound of the formula [II] is used. The reaction can be carried out using a molar mesylating agent (such as mesyl chloride) or a tosylating agent at 0 to 100 ° C. Sulfuric acid: Treatment with sodium is performed in dimethylformamide or dimethylsulfoxide in an inert gas atmosphere such as argon or nitrogen, if necessary, in an amount of 1 to 20 moles per mole of the compound of the formula [II]. The reaction can be carried out at room temperature to a reaction temperature of 150 using sodium sulfide.
このようにして調製した式 [I I I] の化合物の単離は、 通常の糖の分離精製 手段を用いて行なえばよく、 たとえば酢酸ェチルと水で分配後、 シリカゲルカラ ムクロマトグラフィーに付し、 n—へキサン一醉酸ェチルなどの有機溶媒で溶出 することにより分離精製することができる。 第 3工程： The compound of the formula [III] thus prepared may be isolated by a conventional means for separating and purifying a saccharide. For example, after partitioning with ethyl acetate and water, silica gel column chromatography is performed. — Separation and purification can be achieved by elution with an organic solvent such as hexane monoethylate. Third step:
第 3工程は、 式 [I I I] の化合物を酸化剤によりスルホキシド体に導き、 更 に酸無水物で処理することでプンメラー転移を行ない、 式 [IV] で表される化 合物を得る工程である。  In the third step, the compound of the formula [III] is converted to a sulfoxide form with an oxidizing agent, and further treated with an acid anhydride to perform Pummeler rearrangement to obtain a compound of the formula [IV]. is there.
式 [I I I] の化合物のスルホキシドへの誘導は常法に従って行なえばよく、 たとえば、 塩化メチレン中、 アルゴンまたは窒素などの不活性ガス気流下、 -1 00〜0°Cにおいて m—クロ口過安息香酸と処理するか、 あるいはメタノールな どのアルコール系の溶媒中、 メタ過ヨウ素酸ナトリゥムと処理することにより行 なうことができる。  The derivation of the compound of the formula [III] into a sulfoxide may be carried out according to a conventional method. For example, m-chloroperbenzoic acid in methylene chloride under a flow of an inert gas such as argon or nitrogen at -100 to 0 ° C. It can be carried out by treating with an acid or treating with sodium metaperiodate in an alcoholic solvent such as methanol.
また、 プンメラー転移反応も同様に常法に従って行なえばよく、 たとえば、 無 水酢酸などの酸無水物中、 60°C〜還流 で処理することにより式 [IV] の 化^を得ることができる。  The Pummeler rearrangement reaction may be similarly carried out according to a conventional method. For example, a compound represented by the formula [IV] can be obtained by treating in an acid anhydride such as anhydrous acetic acid at 60 ° C. to reflux.
このようにして調製した式 [IV] の化合物の単離は、 通常の糖の分離精製手 段を用いて行なえばよく、 たとえば中和後、 有機溶媒を留去した後、 クロ口ホル ムにより水層より抽出し、 シリカゲルカラムクロマトグラフィ一により分離精製 することができる。 第 4工程:  The compound of the formula [IV] thus prepared may be isolated using a usual sugar separation / purification method. For example, after neutralization, the organic solvent is distilled off, and then the mixture is subjected to chromatography. It can be extracted from the aqueous layer and separated and purified by silica gel column chromatography. Step 4:
第 4工程は、 式 [IV] の化合物にグリコシル化反応により核酸塩基を導入し、 更に糖部保護基を脱保護して式 [I] で表される化合物を得る工程である。  The fourth step is a step of introducing a nucleobase to the compound of the formula [IV] by a glycosylation reaction, and further deprotecting the sugar moiety protecting group to obtain the compound of the formula [I].
グリコシル化反応は、 アルゴンまたは窒素などの不活性ガス気流下、 塩化メチ レン、 クロ口ホルム、 ジクロロェタン、 ァセトニ卜リル、 ジメチルホルムアミ ド などの溶媒中、 式 [IV] の化合物 1モルに対して必要によりシリル化またはァ シルイ匕した核酸塩基 1〜 10モルを用い、 トリメチルシリルトリフルォロメタン スルホネート、 四塩化すず、 四塩ィ匕チタン、 塩ィ kffi鉛、 三フッ化ホウ素などのル イス酸 0. 1〜1 0モルの存在下、 — 5 0〜： L 0 0 °Cで処理することにより実施 することができる。 The glycosylation reaction is carried out under a stream of an inert gas such as argon or nitrogen, in a solvent such as methylene chloride, chloroform, dichloroethane, acetonitrile, or dimethylformamide, relative to 1 mole of the compound of the formula [IV]. If necessary, use 1 to 10 moles of a nucleic acid base which has been silylated or acylated to form a trimethylsilyl trifluoromethanesulfonate, tin tetrachloride, titanium tetrachloride, titanium chloride, kffi lead, boron trifluoride, etc. In the presence of 0.1 to 10 mol of isocyanic acid, —50 to: can be carried out by treating at L 0 ° C.
糖部保護基の脱保護は、 使用した保護基に応じて常法により行なうことができ る。 たとえば、 ベンジル系保護基の場合、 塩ィ匕メチレン中、 アルゴンまたは窒素 などの不活性ガス気流下、 - 1 0 0°C〜室温で三塩ィ匕ホウ素あるいは三臭化ホウ 素で処理することによりベンジル系保護基を脱保護することができる。  Deprotection of the sugar moiety protecting group can be carried out by a conventional method depending on the protecting group used. For example, in the case of a benzyl-based protecting group, it should be treated with boron chloride or boron tribromide at −100 ° C. to room temperature in an inert gas stream such as argon or nitrogen in methylene chloride. Can deprotect the benzyl protecting group.
このようにして得られた L一 4' —チオアラビノフラノヌクレオシド化^!は、 "^のヌクレオシドの単離精製に されている方法を適: M合せて分離精製す ることができる。 たとえば、 溶媒留去後、 シリカゲルカラム、 逆層カラムクロマ トグラフィー、 イオン交換カラムクロマトグラフィー、 活^などの吸着カラム クロマトグラフィーなどにより精製し、 ェタノール等の適当な溶媒から結晶化す れば^く、 必要に応じて: ^として得ることもできる。  The L-1 4'-thioarabinofuranonucleosides ^! Thus obtained can be separated and purified by a method suitable for the isolation and purification of nucleosides of ^. After evaporating the solvent, purify by silica gel column, reverse layer column chromatography, ion exchange column chromatography, adsorption column chromatography such as active column, etc., and crystallize from an appropriate solvent such as ethanol. Depending on: ^ can also be obtained.
(3 ) 本発明の糸賊物 (3) The pirates of the present invention
L一 4' ーチオアラビノフラノヌクレオシド化合物の投与量は、 患者の年齢及 び体重、 疾病、 患者の重篤度、 薬物による忍容性、 投与方法などにより異なり、 これらの条件を総合した上で適宜'^されるものである力 <、 通常 1日当たり 0. 0 0 1〜1 0 0 O m g/k g体重、 好ましくは 0. 0 1〜： L 0 0 m g/k g体重 の範囲内から選ばれ、 一回または 回に分けて投与される。  The dosage of the L-14'-thioarabinofuranonucleoside compound will vary depending on the patient's age and weight, disease, severity of the patient, drug tolerability, administration method, etc. Force, which is appropriately determined in the following <, usually 0.01 to 100 mg / kg body weight per day, preferably 0.01 to: L 0 mg / kg body weight It is administered once or in divided doses.
投与方法は、 経口、 非経口、 経腸、 局所投与などのいずれの経路によっても投 与することができる。  The method of administration can be oral, parenteral, enteral, topical or any other route.
L - 4' —チオアラビノフラノヌクレオシドィ匕合物の製剤化に際しては、 通常 使用される製剤用担体、 賦形剤、 その他の添加剤を用い、 誠物とする。 担体と しては、 乳糖、 カオリン、 ショ糖、 結晶セルロース、 コーンスターチ、 タルク、 寒天、 ぺクチン、 ステアリン酸、 ステアリン酸マグネシウム、 レシチン、 塩化ナ トリウムなどの固体状担体、 グリセリン、 落 油、 ポリビニルピロリ ドン、 ォ リーブ油、 エタノール、 ベンジルアルコール、 プロピレングリコール、 水などの 液状担体を例示することができる。 When formulating the L-4'-thioarabinofuranonucleoside conjugate, the carrier, excipients, and other additives that are ordinarily used for preparation are used to obtain a true product. Carriers include lactose, kaolin, sucrose, crystalline cellulose, corn starch, talc, agar, pectin, stearic acid, magnesium stearate, lecithin, sodium chloride Examples include solid carriers such as thorium, and liquid carriers such as glycerin, oil drop, polyvinylpyrrolidone, olive oil, ethanol, benzyl alcohol, propylene glycol, and water.
剤型としては任意の形態を採ることができ、 たとえば固体状担体を使用する場 合には^ k散剤、 顆粒剤、 カプセル化剤、 座剤、 トローチ剤などを、 液状担体 を使用する場合にはシロップ、 乳液、 軟ゼラチンカプセル、 クリーム、 ゲル、 ぺ 一スト、 スプレー、 注射剤などをそれぞれ例示することができる。 実 施 例  The dosage form can take any form.For example, when a solid carrier is used, a powdery carrier, a granule, an encapsulating agent, a suppository, a troche, and the like are used. Examples include syrup, emulsion, soft gelatin capsule, cream, gel, paint, spray, injection and the like. Example
以下、 本発明を下記の例でもって、 より具体的に説明するが、 本発明はこれら の例によって何等限定されるものではない。  Hereinafter, the present invention will be described more specifically with reference to the following examples, but the present invention is not limited to these examples.
合成例 1.： L-4' ーチオアラビノフラノシルシトシン (式 「1]、 B =シト ン） の合成 Synthesis Example 1: Synthesis of L-4'-thioarabinofuranosylcytosine (Formula [1], B = cytochrome)
(1) 2, 3， 5—トリ一 0—べンジルー D—キシリ トール （式 [I I]、 1^ =ベンジル） の合成  (1) Synthesis of 2,3,5-tri-1 0-benzyl D-xylitol (Formula [II], 1 ^ = benzyl)
D—キシロース 6. 0 gをメタノール 160m 1に懸濁し、 トシル酸 740m gを加えて一 ¾^応させた。 反応後、 トリエチルァミン 1 m iを加えて反応を停 止し、 溶媒を留去した。 残渣をシリカゲルカラムクロマトグラフィーにより精製 し、 1一メチル体 （6. 33 g, 96%) を得た。  6.0 g of D-xylose was suspended in 160 ml of methanol, and 740 mg of tosylic acid was added thereto for reaction. After the reaction, the reaction was stopped by adding 1 mi of triethylamine, and the solvent was distilled off. The residue was purified by silica gel column chromatography to obtain 1-methyl form (6.33 g, 96%).
1—メチル体 6. 33 gを DMF 200mlに溶かし、 アルゴン気流下、 0°C で 60%ナトリゥムハイドライド 5. 4 gを加え、 更にべンジルクロライド 17. 7m 1を滴下した。 滴下後、 室温に戻してー 応させた。 反応後、 氷水に反応 液を加えて反応を停止した。 エーテルを加えて分配し、 有機層を水で 3回洗った 後、 飽和食塩水で洗った。 有機層を芒硝で乾燥した後、 溶媒を留去し、 残渣をシ リカゲルカラムにより精製し、 トリベンジル体 （10. 2 g, 61%) を得た。 トリベンジル体 9. 16 gを THF 75m 1に溶かし、 6 N塩酸 15 Omlを 加えて室温で 3日間反応させた。 反応後、 炭酸水素ナトリゥムを加えて中和し、 有機層を分液して得た。 得られた有機層の溶媒を留去し、 残渣を酢酸ェチルで溶 かし、水 3回と飽和: ^水で洗った。 有機層を芒硝乾燥した後、 溶媒を留去した。 残澄を T H F 120 m 1に溶かし、水素化ホゥ素リチウム 460mgを加えて 2 時間反応させた。 酔酸で中和した後、 溶媒を留去した。 残渣を酢酸ェチルで溶か し、 有 を水 3回と飽和食塩水で洗って芒硝で乾燥した。 溶媒を留去した後、 シリカゲルカラムで精製し、 題記化合物 （5. 84 g, 66%) を得た。 6.33 g of the 1-methyl compound was dissolved in 200 ml of DMF, 5.4 g of 60% sodium hydride was added at 0 ° C. in an argon stream, and 17.7 ml of benzyl chloride was further added dropwise. After the dropwise addition, the mixture was returned to room temperature and reacted. After the reaction, the reaction solution was added to ice water to stop the reaction. Ether was added and the mixture was partitioned, and the organic layer was washed three times with water and then with a saturated saline solution. After the organic layer was dried over sodium sulfate, the solvent was distilled off, and the residue was purified by a silica gel column to obtain a tribenzyl form (10.2 g, 61%). 9.16 g of the tribenzyl compound was dissolved in 75 ml of THF, 15 Oml of 6 N hydrochloric acid was added, and the mixture was reacted at room temperature for 3 days. After the reaction, the mixture was neutralized by adding sodium hydrogen carbonate, and the organic layer was separated. The solvent of the obtained organic layer was distilled off, the residue was dissolved with ethyl acetate, and the mixture was washed three times with water and washed with water. After the organic layer was dried over sodium sulfate, the solvent was distilled off. The residue was dissolved in 120 ml of THF, 460 mg of lithium borohydride was added, and the mixture was reacted for 2 hours. After neutralization with carboxylic acid, the solvent was distilled off. The residue was dissolved in ethyl acetate, and the residue was washed three times with water and saturated saline, and dried over sodium sulfate. After evaporating the solvent, the residue was purified by a silica gel column to obtain the title compound (5.84 g, 66%).
1 H-NMR (CDC ) δ Ί . 36-7. 22 (15 Η, m) , 4. 68- 4. 43 (6H, m)， 4. 07-4. 03 (1H， m) , 3. 83-3. 75 1 H-NMR (CDC) δ Ί. 36-7. 22 (15 Η, m), 4. 68- 4. 43 (6H, m), 4. 07-4. 03 (1H, m), 3. 83-3. 75
(2H， m) ， 3. 72 (1H, dd， J =2. 0， 5. 9 H z) , 3. 67(2H, m), 3.72 (1H, dd, J = 2.0, 5.9 Hz), 3.67
(1H， d t, J = 3. 9, 5. 9Hz) ， 3. 51 (1H， d d， J = 6. 4, 9. 3Hz) ， 3. 42 (1H, d d， J = 6. 4, 9. 3Hz) , 2. 81(1H, dt, J = 3.9, 5.9Hz), 3.51 (1H, dd, J = 6.4, 9.3Hz), 3.42 (1H, dd, J = 6.4, 9 .3Hz), 2.81
(1H， b r d， D2 0 exchange ab l e, J =5. 4Hz) , 2. 57 (1H, b r, 0 exchangeab l e) (1H, brd, D 2 0 exchange ab le, J = 5.4 Hz), 2.57 (1H, br, 0 exchange ab le)
FAB-MS m/z : 423 (M+ +H) FAB-MS m / z: 423 (M + + H)
元素分析 (C26H3()05 ·Η2 0として） Elemental analysis (as C 26 H 3 () 0 5 · Η 20 )
計算値： C, 70. 89 ; Η, 7. 32  Calculated: C, 70.89; Η, 7.32
分析値： C, 71. 22 ; Η, 7. 01  Analytical values: C, 71. 22; Η, 7.01
Lai D =-12. 7 (c 1. 1, CHC ） Lai D = -12. 7 (c 1.1, CHC)
(2) 1, 4—アンヒドロー 2, 3, 5—トリー 0—ベンジル一 4—チォー D —ァラビトール （式 [I I I] 、 1^ =ベンジル） の合成 (2) Synthesis of 1,4-anhydro-2,3,5-tree 0-benzyl-14-thio D-arabitol (Formula [III], 1 ^ = benzyl)
2, 3, 5—トリー 0—ベンジルー D—キシリ トール （式 [I I] 、 1^ =ベ ンジル） 5. 22 gをピリジン 60 m 1に溶かし、 ァルゴン気流下、 0 °Cでメ夕 ンスルホニルクロリ ド 2. 88m 1を滴下した。 滴下後、 室温に戻して 1. 5時 間撹拌した。 氷水を加えて反応を停止し、 溶媒を留去した。 残渣を酢酸ェチルに 溶かし、水で 3回と飽和^水で洗って有機層を乾燥させた。 溶媒を留去し、 残 渣を DMF 100m lに溶かし、 硫化ナトリゥム 9水和物 8. 93 gを加えて 1 00°Cで 3時間反応させた。 反応後、 室温に戻し、 溶媒を留去した後、 鲊酸ェチ ルに溶かし、 水で 3回と飽和^水で洗って、 有機層を芒硝で乾燥した。 溶媒を 留去した後、 シリカゲルカラムで精製し、 題記化合物 （4. 38 g, 84%) を 得た。 2,3,5-tree 0-benzyl-D-xylitol (Formula [II], 1 ^ = 5.22 g was dissolved in 60 ml of pyridine, and 2.88 ml of malesulfonyl chloride was added dropwise at 0 ° C under a stream of argon. After the dropwise addition, the mixture was returned to room temperature and stirred for 1.5 hours. The reaction was stopped by adding ice water, and the solvent was distilled off. The residue was dissolved in ethyl acetate and washed with water three times and saturated water, and the organic layer was dried. The solvent was distilled off, the residue was dissolved in 100 ml of DMF, 8.93 g of sodium sulfide nonahydrate was added, and the mixture was reacted at 100 ° C. for 3 hours. After the reaction, the temperature was returned to room temperature, the solvent was distilled off, the residue was dissolved in ethyl acetate, washed with water three times and saturated water, and the organic layer was dried over sodium sulfate. After evaporating the solvent, the residue was purified by a silica gel column to obtain the title compound (4.38 g, 84%).
1 H-NMR (CDC 13 ) 61. 35-7. 27 (15 H， m) ， 4. 61 (2H, s) , 4. 55-4. 46 (4H, m) , 4. 19 (1Η, q, J =4. 4Hz) ， 4. 11 (1H, t, J = 3. 9Hz) , 3. 69 (1 H, d d, J =7. 8, 8. 8Hz) ， 3. 56 (1H, d d d, J = 3. 9, 6. 4, 7. 8Hz) ， 3. 50 (1 H, dd, J = 5. 9, 8. 8Hz) , 3. 08 (1H， dd, J = 4. 9, 11. 2Hz) , 2. 90 (1H, d d, J =4. 4, 11. 2Hz) 1 H-NMR (CDC 1 3 ) 61. 35-7. 27 (15 H, m), 4. 61 (2H, s), 4. 55-4. 46 (4H, m), 4. 19 (1Η , q, J = 4.4 Hz), 4.11 (1H, t, J = 3.9 Hz), 3.69 (1 H, dd, J = 7.8, 8.8 Hz), 3.56 (1H , ddd, J = 3.9, 6.4, 7.8 Hz), 3.50 (1 H, dd, J = 5.9, 8.8 Hz), 3.08 (1H, dd, J = 4. 9, 11.2Hz), 2.90 (1H, dd, J = 4, 4, 11.2Hz)
FAB-MS m/ z : 421 (M+ +H) FAB-MS m / z: 421 (M + + H)
元素分析 （C26H28〇3 Sとして） Elemental analysis (as C 26 H 28 〇 3 S)
理論値： C, 74. 25 ; H, 6. 71  Theoretical: C, 74.25; H, 6.71
分析値： C, 74. 17 ; H, 6. 75  Analytical values: C, 74.17; H, 6.75
[α] D = 一 0. 38 (c 2. 5， CHC 13 ) [alpha] D = one 0. 38 (c 2. 5, CHC 1 3)
(3) 1—0ァセチルー 2， 3, 5—トリー 0—ベンジルー 4一チォー D—ァラ ビノース （式 [I V]、 R1 =ベンジル、 R2 =ァセチル） の合成 1, 4—アンヒドロー 2, 3, 5 _トリ一 0—ベンジル一 4ーチォ一D—ァラ ビトール （式 [I I I] 、 1^ =ベンジル） 3. 43 gを塩ィ匕メチレン 4 Om 1 に溶かし、 アルゴン気流下、 — 78°Cで m—クロ口過安息香酸 1. 76gを含ん だ塩化メチレン溶液 （40ml) を滴下した。 滴下後、 一 78°Cで 30分間撹拌 した。 飽和炭酸水素ナトリウム溶液を加えて反応を停止し、 クロ口ホルムで 3回 抽出した。 有機層を 10%チォ硫酸ナトリゥム溶液、 飽和炭酸水素ナトリウム溶 液、 飽和: ^水で分配した後、 芒硝で乾燥した。 溶媒を留去し、 残渣を無水酢酸 4 Omlに溶解し、 3時間、 100°Cに保った。 溶媒を留去した後、 トルエンで 3回共沸した。 残渣を酢酸ェチルに溶かし、 飽和炭酸ナトリウム溶液で 3回と飽 和: ^水で洗って有,を乾燥させた。 溶媒を留去し、 残渣をシリカゲルカラム で精製し、 題記化合物 （1. 94g, 50%) を得た。 (3) Synthesis of 1-0 acetyl-2,3,5-tree 0-benzyl-4 monothio D-arabinose (Formula [IV], R 1 = benzyl, R 2 = acetyl) 1,4-Anhydro-2,3,5_tri-1 0-benzyl-1-thio-1D-arabitol (Formula [III], 1 ^ = benzyl) 3.43 g is dissolved in Shioiden methylene 4 Om 1 In a stream of argon, at −78 ° C., a methylene chloride solution (40 ml) containing 1.76 g of m-chloroperbenzoic acid was added dropwise. After the dropwise addition, the mixture was stirred at 178 ° C for 30 minutes. The reaction was quenched by the addition of a saturated sodium hydrogen carbonate solution, and the mixture was extracted three times with chloroform. The organic layer was partitioned between a 10% sodium thiosulfate solution, a saturated sodium bicarbonate solution, and saturated: ^ water, and then dried over sodium sulfate. The solvent was distilled off, and the residue was dissolved in acetic anhydride (4 Oml) and kept at 100 ° C for 3 hours. After the solvent was distilled off, the residue was azeotroped with toluene three times. The residue was dissolved in ethyl acetate and saturated with saturated sodium carbonate solution three times: ^ Washed with water and dried. The solvent was distilled off, and the residue was purified by a silica gel column to give the title compound (1.94 g, 50%).
1 H-NMR (CDC ) 67. 35- 7. 24 (15 Η, m) ， 6. 07 (0. 7H, d, J = 3. 9Hz) , 5. 98 (0. 3 H, d， J =2. 9Hz) ， 4. 83— 4. 48 (6H, m)， 4. 26 (0. 3 H, d d, J = 2. 9, 5. 4Hz) ， 4. 18 (0. 7H, d d， J =4. 4, 8. 8Hz) ， 4. 1 2 (0. 7H, d d, J =6. 4， 8. 8Hz) ， 4. 03 (0, 3 H， dd, J = 5. 4， 6. 4Hz) , 3. 77 (0. 3 H, q, J =6. 4Hz) ， 3. 71 (0. 3H, dd, J = 5. 9， 9. 3Hz) , 3. 68 (0. 7 H, d d， J = 5. 9， 9. 3Hz) ， 3. 51 (0. 7 H, d d, J = 6. 8， 9. 3H z)， 3. 46 (0. 3H, dd， J = 6. 4, 9. 3Hz) , 3. 40 (0. 7H, q, J = 6. 4Hz) , 2. 06 (3H, s) 1 H-NMR (CDC) 67.35-7.24 (15 Η, m), 6.07 (0.7 H, d, J = 3.9 Hz), 5.98 (0.3 H, d, J = 2.9 Hz), 4.83—4.48 (6H, m), 4.26 (0.3 H, dd, J = 2.9,5.4 Hz), 4.18 (0.7H, dd) , J = 4.4, 8.8 Hz), 4.12 (0.7 H, dd, J = 6.4, 8.8 Hz), 4.03 (0, 3 H, dd, J = 5.4) , 6.4Hz), 3.77 (0.3H, q, J = 6.4Hz), 3.71 (0.3H, dd, J = 5.9, 9.3Hz), 3.68 (0 7 H, dd, J = 5.9, 9.3 Hz), 3.51 (0.7 H, dd, J = 6.8, 9.3 Hz), 3.46 (0.3 H, dd, J = 6.4, 9.3Hz), 3.40 (0.7H, q, J = 6.4Hz), 2.06 (3H, s)
FAB-MS m/z : 419 (M+ - OAc) FAB-MS m / z: 419 (M + -OAc)
元素分析 （C28H3()05 S · 0. 25H2 0として） Elemental analysis (as C 28 H 3 () 0 5 S · 0.25H 20 )
理論値： C, 69. 61 ; H, 6. 36 分析値： C, 69. 42 ; H, 6. 26 Theoretical: C, 69.61; H, 6.36 Analytical values: C, 69.42; H, 6.26
[ ] D = +29. 8 (c 2. 0, CHC )  [] D = +29.8 (c 2.0, CHC)
(4) L-4' ーチオアラビノフラノシルシトシン (式 [I] 、 Β-シトシ:^ の合成 (4) L-4'-Thioarabinofuranosylcytosine (Synthesis of formula [I], Β-cytosine: ^
N4—ァセチルシトシン 459mgをァセトニトリル 10m 1に溶解し、 B S A 860 ^ 1を加えアルゴン気流下、 5. 5時間還流した。 減圧下溶媒を留去後、 残渣をァセトニトリル 5 m 1に溶解し、 1—0ァセチルー 2， 3， 5—トリー 0 —ベンジル一 4—チォー D—ァラビノース （式 [IV] 、 R1 =ベンジル、 R2 =ァセチル） 479mgを加えた。 この溶液に TMSトリフレート 290 1を 加え、 室温で 1. 5時間 »した。 飽和炭酸水素ナトリゥム溶液を加えて反応を 停止し、 不溶物をセライ卜ろ過した。 ろ液をクロ口ホルムで 3回抽出し、 有^ を乾燥した。 濃縮した後、 シリカゲルカラムにより精製を行ない、 ヌクレオシド ィ匕合物 445mg (78%) を得た。 459 mg of N4-acetylcytosine was dissolved in 10 ml of acetonitrile, BSA 860 ^ 1 was added, and the mixture was refluxed for 5.5 hours under an argon stream. After evaporating the solvent under reduced pressure, the residue was dissolved in 5 ml of acetonitrile, and 1-0 acetyl-2,3,5-tree 0-benzyl-14-thio D-arabinose (Formula [IV], R 1 = benzyl, (R 2 = acetyl) 479 mg was added. TMS triflate 2901 was added to this solution, and the mixture was kept at room temperature for 1.5 hours. The reaction was stopped by adding a saturated sodium hydrogen carbonate solution, and the insolubles were filtered through celite. The filtrate was extracted three times with a black hole form, and dried. After concentration, purification was carried out using a silica gel column to obtain 445 mg (78%) of a nucleoside conjugate.
ヌクレオシド化合物 417mgを塩化メチレン 1 Omlに溶解し、 一78 に 冷却した。 この溶液に 1M三塩ィ匕ホウ雜化メチレン溶液 4. 38mlを滴下し、 一 78°Cで 30分間撹拌した。 一 20°Cに昇温し、 更に 3時間撹拌した後、 飽和 炭酸水素ナトリウム溶液を加えて反応を停止した。 溶媒を留去し、 残渣にメタノ ール 5ml、 濃アンモニア水 5 mlを加え、 室温で一晚撹拌した。 溶媒を留去し、 残渣をエタノールで 3回共沸し、 シリカゲルカラムにより精製した。 得られたァ ノマー混合物を OD Sカラムクロマトにより精製並びにァノマーの分離を行ない、 題記化^!の ーァノマー （64mg， 34%)及び) S—ァノマー （36mg, 19%) を得た。 The nucleoside compound (417 mg) was dissolved in methylene chloride (1 Oml) and cooled to 178. To this solution, 4.38 ml of a 1M methylene chloride solution was added dropwise and stirred at 178 ° C for 30 minutes. The temperature was raised to 120 ° C, and the mixture was further stirred for 3 hours, and the reaction was stopped by adding a saturated sodium hydrogen carbonate solution. The solvent was distilled off, 5 ml of methanol and 5 ml of concentrated aqueous ammonia were added to the residue, and the mixture was stirred at room temperature for 10 minutes. The solvent was distilled off, and the residue was azeotroped with ethanol three times, and purified by a silica gel column. The obtained anomeric mixture was purified by ODS column chromatography and the anomers were separated to obtain the enantiomer of the title ^! (64 mg, 34%) and the) S-anomer (36 mg, 19%).
S—ァノマー ： 1 H- NMR (DMSO-dg ) <57. 96 (1 H， d, J = 7. 8Hz) , 7. 10， 7. 01 (t o t a l 2 H, b r s, Dg 0 exchangeab l e) , 6. 33 (1H, d， J = 4. 9Hz) , 5. 69 (1H, d, J =7. 8H z) , 5. 56 (1 H, d， J = 4. 9H z, D2 0 exchangea b 1 e) , 5. 35 (1 H, d, J = 3. 9Hz, D2 0 exchange a b 1 e)， 5. 05 (1 H, t, J = 5. 4Hz, D2 0 exchangea b l e) ， 3. 98 -3. 92 (2H， m) , 3. 78 (1H, d t, J = 5. 4， 11. 2Hz) , 3. 58 (1H, d t, J = 5. 9， 11. 2Hz) , 3. 18-3. 13 (1H， m) S—Anomer: 1 H-NMR (DMSO-dg) <57.96 (1 H, d, J = 7.8 Hz), 7.10, 7.01 (total 2 H, brs, D g 0 exchangeab le), 6.33 (1H, d, J = 4. 9Hz), 5. 69 (1H, d, J = 7. 8H z), 5. 56 (1 H, d, J = 4. 9H z, D 2 0 exchangea b 1 e), 5.35 (1 H, d, J = 3.9 Hz, D 20 exchange ab 1 e), 5.05 (1 H, t, J = 5.4 Hz, D 20 exchange a ble), 3 98 -3.92 (2H, m), 3.78 (1H, dt, J = 5.4, 11.2Hz), 3.58 (1H, dt, J = 5.9, 11.2Hz), 3. 18-3. 13 (1H, m)
FAB MS m/z : 260 (M + H+ ) FAB MS m / z: 260 (M + H + )
元素分析 （C9 H13N3 04 S · 0. 5H2 0として） Elemental analysis (as C 9 H 13 N 3 0 4 S · 0. 5H 2 0)
理論値： C， 40. 29 ; H, 5. 26 ; N, 15. 66  Theoretical: C, 40.29; H, 5.26; N, 15.66
分析値： C， 40. 22 ; H, 5. 06 ; N, 15. 38  Analytical values: C, 40.22; H, 5.06; N, 15.38
a—ァノマー： a—anomer:
1 H-NMR (DMSO-dg ) 57. 89 (1H， d， J = 7. 3Hz) , 7. 14, 7. 08 (t o t a l 2 H, b r s, D2 0 exchangeab l e)， 5. 85 (1H, d, J = 7. 3Hz) , 5. 76 (1H， d, J = 7. 3Hz) , 5. 57 (1 H, d， J = 5. 9H z, D9 0 exchangea b 1 e) , 5. 44 (1 H, d， J =4. 9Hz, D 0 exchangea b 1 e) , 4. 87 (1 H, t, J =5. 4Hz, D0 0 exchangea b l e) , 3. 93 (1H, q, J =6. 8Hz) , 3. 82 (1H， d t, J =4. 4, 10. 7Hz) , 3. 64 (1H, d t, J =4. 9， 7. 3Hz) , 3. 45 (1H, d t, J =3. 9， 7. 8Hz) , 3. 36 (1H， d d d, J =5. 9, 8. 3, 10. 7Hz) 1 H-NMR (DMSO-dg) 57.89 (1H, d, J = 7.3 Hz), 7.14, 7.08 (total 2 H, brs, D 20 exchangeable), 5.85 (1H , d, J = 7. 3Hz) , 5. 76 (1H, d, J = 7. 3Hz), 5. 57 (1 H, d, J = 5. 9H z, D 9 0 exchangea b 1 e), 5.44 (1 H, d, J = 4.9 Hz, D 0 exchangea b 1 e), 4.87 (1 H, t, J = 5.4 Hz, D 0 0 exchangea ble), 3.93 (1H , q, J = 6.8 Hz), 3.82 (1H, dt, J = 4.4, 10.7 Hz), 3.64 (1H, dt, J = 4.9, 7.3 Hz), 3. 45 (1H, dt, J = 3.9, 7.8Hz), 3.36 (1H, ddd, J = 5.9, 8.3, 10.7Hz)
FAB MS m/z : 260 (M+H+ ) 元素分析 （C9 H13 3 04 S · 0. 25H2 0として） FAB MS m / z: 260 (M + H +) Elemental analysis (as C 9 H 13 3 0 4 S · 0. 25H 2 0)
理論値： C, 40. 98 ; H, 5. 16 ; N, 15. 93  Theory: C, 40.98; H, 5.16; N, 15.93
分析値： C， 41. 01 ; H， 5. 04 ; N, 15. 82  Analytical values: C, 41.01; H, 5.04; N, 15.82
合成例 2 : L— 4' —チオアラビノフラノシルチミン (式 [I]、 8=チミン） の合成 Synthesis Example 2: Synthesis of L-4'-thioarabinofuranosyl thymine (Formula [I], 8 = thymine)
4—ァセチルシトシンの代わりにチミンを用い、 上記と同様の方法により題 記化合物 ァノマ一 （64mg, 34%)及び /3—ァノマー （56mg， 19 %) を得た。  Using thymine instead of 4-acetylcytosine, the title compound Panoma-1 (64 mg, 34%) and the / 3-anomer (56 mg, 19%) were obtained in the same manner as above.
/3—ァノマー  / 3—anomer
H-NMR (DMS 0-d„ ) δ 25 (1Η, s e x c h a  H-NMR (DMS 0-d „) δ 25 (1Η, se x c h a
D2 0 D 2 0
nge ab l e) , 7. 93 (1H， d， J = 1. 0Hz) ， 6. 07 (1H, d, J = 5. 9Hz) ， 5. 69 (1 H, d， J =5. 4Hz, D2 0 exc hangeab l e) , 5. 40 (1 H, d, J = 4. 9Hz, 0 exc hangeab l e) , 5. 21 (1H, t, J = 5. 1Hz, D9 0 exc hangeab l e) , 4. 00 (1H, q, J = 5. 9Hz) ， 3. 94 (1nge ab le), 7.93 (1H, d, J = 1.0Hz), 6.07 (1H, d, J = 5.9Hz), 5.69 (1H, d, J = 5.4Hz, D 2 0 exc hangeab le), 5.40 (1 H, d, J = 4.9 Hz, 0 exc hangeab le), 5.21 (1H, t, J = 5.1 Hz, D 9 0 exc hangeab le) , 4.00 (1H, q, J = 5.9 Hz), 3.94 (1
H， q, J =5. 4Hz) , 3. 75 (1H, d t, J =4. 9， 11. 2Hz) ， 3. 66 (1H, d t, J = 5. 9， 11. 2Hz) , 3. 13 (1H， q,H, q, J = 5.4 Hz), 3.75 (1H, dt, J = 4.9, 11.2 Hz), 3.66 (1H, dt, J = 5.9, 11.2 Hz), 3 . 13 (1H, q,
=5. 4Hz)， 77 (3H, s) = 5.4 Hz), 77 (3H, s)
+  +
FAB MS m/z : 275 (M + H' )  FAB MS m / z: 275 (M + H ')
元素分析 （C10H14N2 05 Sとして） Elemental analysis (as C 10 H 14 N 2 0 5 S)
理論値： C， 43. 79 ; H, 5. 14 Ν, 10. 21  Theoretical, C, 43.79; H, 5.14 ,, 10.21
分析値： C, 43. 64； H, 5. 31 Ν, 10. 20  Analytical values: C, 43.64; H, 5.31Ν, 10.20
α—ァノマー： α-anomers:
H-NMR (DMS O-dg ) <511. 27 ( 1 H, s , Dn 0 excha n g e a b 1 e) , 7. 84 (1 H， s) ， 5. 74 (1 H， d, J = 7. 8H z) ， 5. 67 (1 H, d, J = 5. 7Hz, D2 0 e x c h a n g e a b l e) , 5. 52 (1 H, d, J =4. 9 H z, 0 e x c h a n g e a b l e) ， 4. 89 (1H, t, J = 5. 1 H z, D2 0 e x c h a n g e a b l e) ， 3. 99 (1 H， d t， J =5. 7, 7. 8H z) , 3. 87-3. 82 (1 H, m) , 3. 60 (1 H, d t, J = 4. 9， 8. 3 H z) ， 3. 52 (1 H, d t， J = 3. 4, 8. 3H z) ， 3. 40 -3. 35 (1 H, m) , 1. 81 (3H, s) H-NMR (DMS O-dg) <511.27 (1 H, s, D n 0 excha ngeab 1 e), 7.84 (1 H, s), 5.74 (1 H, d, J = 7.8 Hz), 5.67 (1 H, d, J = 5.7 Hz, D 20 exchangeable), 5.52 (1 H, d, J = 4.9 Hz, 0 exchangeable), 4.89 (1H, t, J = 5.1 Hz, D 2 0 exchangeable), 3.99 ( 1 H, dt, J = 5.7, 7.8H z), 3.87-3.82 (1 H, m), 3.60 (1 H, dt, J = 4.9, 8.3 H z), 3.52 (1 H, dt, J = 3.4, 8.3 Hz), 3.40 -3. 35 (1 H, m), 1.81 (3H, s)
FAB MS m/z : 275 (M + H+ ) FAB MS m / z: 275 (M + H + )
元素分析 (C1()H14N2 05 Sとして） Elemental analysis (C 1 () as H 14 N 2 0 5 S)
理論値： C, 43. 79 ; H, 5. 14 ; N， 10. 21  Theoretical: C, 43.79; H, 5.14; N, 10.21
分析値： C, 43. 50 ; H, 5. 10 ； N， 9. 82 合成例 3 ： —チオアラビノフラノシルアデニン （式 [I] 、 B=アデ二 ン） の合成  Analytical values: C, 43.50; H, 5.10; N, 9.82 Synthesis example 3: Synthesis of thioarabinofuranosyladenine (Formula [I], B = adenyl)
N 4—ァセチルシトシンの代わりにアデニンを用い、 上記と同様の方法により題 記化合物なーァノマー （16mg) 及び^ーァノマー （1 06m g, c r u d e) を得た。  Adenine (16 mg) and ^ anomer (106 mg, crude) as the title compounds were obtained in the same manner as described above, except that adenine was used instead of N 4 -acetylcytosine.
β— Ύゾマー ： β-Ύsomer:
H-NMR (DMSO-d„ ) δ 8. 36 (1 H， s) , 8. 3 (1 H， s)H-NMR (DMSO-d „) δ 8.36 (1 H, s), 8.3 (1 H, s)
， 7. 22 (2H， b r， D 0 e x c h a n g e a b l e) , 6. 06 (1 H， d, J =4. 9H z) , 5. 77 (1H, d, J =4. 9H z, D2 0 e x c h a n g e a b l e) , 5. 58 (1 H, d， J =4. 4H z, 0 e x c h a n g e a b l e) , 5. 24 (1 H, J = 5. 4H z, D2 0 e xchange ab l e) , 4. 16-4. 10 (2H, m)， 3. 85 (1H, d t, J = 4. 9, 10. 7Hz) , 3. 73 (1H， d t, J = 5. 9, 10. 7Hz)， 3. 28-3. 24 (1H, m) , 7. 22 (2H, br, D 0 exchangeable), 6. 06 (1 H, d, J = 4. 9H z), 5. 77 (1H, d, J = 4. 9H z, D 2 0 exchangeable ), 5.58 (1 H, d, J = 4.4 Hz, 0 exchangeable), 5.24 (1 H, J = 5.4 Hz, D 2 0 e xchange ab)), 4.16-4.10 (2H, m), 3.85 (1H, dt, J = 4.9, 10.7 Hz), 3.73 (1H, dt, J = 5.9 , 10.7Hz), 3.28-3.24 (1H, m)
FAB MS m/z : 284 (M + H+ )  FAB MS m / z: 284 (M + H +)
α—ァノマー：  α-anomers:
1 H-NMR (DMSO-dg ) δ 8. 40 (1H, s) ， 8. 15 (1H， s) 1 H-NMR (DMSO-dg) δ 8.40 (1H, s), 8.15 (1H, s)
， 7. 24 (2 H, b r， 0 exchange ab l e) , 5. 80 (1, 7.24 (2 H, b r, 0 exchange ab le), 5.80 (1
H， b r, D2 0 exchange ab l e) , 5. 73 (1 H, d, J = 7.H, br, D 2 0 exchange ab le), 5.73 (1 H, d, J = 7.
3H z) , 5. 63 (1H, b r, 0 exchange ab l e) , 4.3H z), 5.63 (1H, b r, 0 exchange ab l e), 4.
93 (1H, b r, D2 0 exchange ab l e) , 4. 56 (1 H, t,93 (1H, br, D 2 0 exchange ab le), 4.56 (1 H, t,
J = 7. 3Hz) , 3. 89 (1H, d d, J =3. 4, 10. 7Hz) ， 3.J = 7.3Hz), 3.89 (1H, d d, J = 3.4, 10.7Hz), 3.
74 (1H， t, J = 7. 8Hz) ， 3. 66 (1H, d t, J = 3. 9, 7.74 (1H, t, J = 7.8 Hz), 3.66 (1H, dt, J = 3.9, 7.
8Hz) , 3. 43 (1H， d d， J =7. 8， 10. 7Hz) 8Hz), 3.43 (1H, dd, J = 7.8, 10.7Hz)
FAB MS m/z : 284 (M + H+ )  FAB MS m / z: 284 (M + H +)
元素分析（C10H13NF 03 S · 0. 5H2 0 Elemental analysis (C 10 H 13 N F 0 3 S · 0.5 H 2 0
理論値： C, 41. 09 ; H, 4. 83 ; N, 23. 96  Theory: C, 41.09; H, 4.83; N, 23.96
分析値： C， 41. 41 ; H， 4. 56 ; N, 23. 64 成例 4： 2, 6—ジアミノー (L-4/ —チオアラピノフラノシル) プリ上 (式 [I] 、 B = 2， 6—ジァミノプリン） の合成 Analytical values: C, 41.41; H, 4.56; N, 23.64 Example 4: 2,6-diamino- (L-4 / —thioarapinofuranosyl) on pri (formula [I], B = 2, 6—diaminopurine)
N 4—ァセチルシ卜シンの代わりに 2， 6—ジァミノプリンを用い、 上記と同様 の方法により題記化合物 ーァノマー （82mg, 23%) 及び^ーァノマー (44mg, 12%) を得た。  Using 2,6-diaminopurine in place of N4-acetylcytosine, the title compound -anomer (82 mg, 23%) and ^ -anomer (44 mg, 12%) were obtained in the same manner as above.
β—了ゾマー ： 1 H-NMR (DMS O-dg ) δ 7. 91 (I H, s) ， 6. 64 (2H， b r, D2 0 e x c h a n g e a b l e) , 5. 93 (1 H, d， J = 5. 4H z) , 5. 76 (2H， b r， D2 0 e x c h a n g e a b l e) , 5. 70 (I H, d， J = 5. 4H z， D2 0 e x c h a n g e a b l e) , 5. 45 (I H, d， J =4. 9H z, D2 0 e x c h a n g e a b l e) , 5. 12 (1 H, t， J = 5. 4H z, D2 0 e x c h a n g e a b l e) , 4. 1 1 (1 H， q， J =5. 4H z) ， 4. 02 (I H, q, J =5. 9Hz) , 3.β—Ryosomer: 1 H-NMR (DMS O- dg) δ 7. 91 (IH, s), 6. 64 (2H, br, D 2 0 exchangeable), 5. 93 (1 H, d, J = 5. 4H z) , 5. 76 (2H, br, D 2 0 exchangeable), 5. 70 (IH, d, J = 5. 4H z, D 2 0 exchangeable), 5. 45 (IH, d, J = 4. 9H z , D 20 exchangeable), 5.12 (1 H, t, J = 5.4 Hz, D 20 exchangeable), 4.1 1 (1 H, q, J = 5.4 Hz), 4.02 (IH, q, J = 5.9Hz), 3.
83 (1 H， d t， J = 5. 4, 10. 7H z) , 3. 66 (I H, d t， J =83 (1 H, dt, J = 5.4, 10.7 Hz), 3.66 (I H, dt, J =
5. 9, 1 0. 7H z) ， 3. 23-3. 1 9 (I H, m) 5. 9, 10.7Hz), 3.23-3.19 (I H, m)
FAB MS m/z : 299 (M + H+ )  FAB MS m / z: 299 (M + H +)
a—ァノマー： a—anomer:
1 H-NMR (DMSO-dg ) δ 7. 99 (I H, s) ， 6. 66 (2H, b r, D2 0 e x c h a n g e a b l e) , 5. 76 (2 H, b r, 0 e x c h a n g e a b l e) , 5. 75 (1 H, d, J =5. 9H z， D2 0 e x c h a n g e a b l e) , 5. 57 (1 H, d， J =5. 4H z, D2 0 e x c h a n g e a b l e) , 5. 55 (1 H， d， J =7. 3H z) ， 4. 89 (I H, t, J = 5. 4H z, D2 0 e x c h a n g e a b l e) , 4. 42 (I H, q, J = 7. 3H z) , 3. 85 (I H, d t, J =4. 4, 1 0. 7 1 H-NMR (DMSO-dg) δ 7.99 (IH, s), 6.66 (2H, br, D 20 exchangeable), 5.76 (2 H, br, 0 exchangeable), 5.75 ( 1 H, d, J = 5.9 Hz, D 20 exchangeable), 5.57 (1 H, d, J = 5.4 Hz, D 20 exchangeable), 5.55 (1 H, d, J = 7. 3H z), 4. 89 (IH, t, J = 5. 4H z, D 2 0 exchangeable), 4. 42 (IH, q, J = 7. 3H z), 3. 85 (IH, dt, J = 4, 4, 1 0.7
Hz) , 3. 69 (I H, d t， J = 4. 9, 7. 8H z) ， 3. 57 (I H, d t, J = 3. 9， 7. 8H z) , 3. 40 (1 H， d d d, Hz), 3.69 (IH, dt, J = 4.9, 7.8Hz), 3.57 (IH, dt, J = 3.9, 7.8Hz), 3.40 (1H, ddd,
J = 5. 9， 7. 8， 1 0. 7H z)  J = 5.9, 7.8, 10.7Hz)
FAB MS m/z : 299 (M+H+ ) 試験例 FAB MS m / z: 299 (M + H +) Test example
1) インビトロ抗 B型肝炎ウィルス （HBV) 活性：  1) In vitro anti-hepatitis B virus (HBV) activity:
(1) 細胞  (1) cells
HBV遺^?が導入されたヒト肝ガン細胞株、 HB 6 1 1 (Proc. Natl. Acad. Sci. U.S.A. 84, 444-448(1987) ) を用い、 周藤らの方法 (Microbiol. Immunol. ,40(2) , 153-159(1996) ) に若干の変更を加え、 薬剤の抗 Η Β V活性を測定し た。 すなわち、 ΗΒ 6 1 1細胞を 1 0%牛胎 i¾fil清、 1 00 1 UZm lベニシリ G、 1 00 g/m 1ス卜レプトマイシンおよび lmg/m 1ゲネチシンを含 むダルベッコ変法 MEMに懸濁し、 96穴マルチウエルプレートに 1ゥエル当た り 20000個 ¾した。炭酸ガスインキュベータ一内で 37°C、 3日間培養し、 細胞を集密的 （confluent ) に生育させた後、 試験に供した。 試験薬剤を通常 1 00 /i g/in lの濃度から 10倍段階希釈で 4段階、 培地で希釈して細胞に加え、 培養した。 薬剤を含む培地で 2日^ &び 4日後に培地交換を行なった。 対照とす る細胞は、 薬剤を含まない培地で培養した。 7日後に各ゥエルから培養上清を回 収し、 別の 96穴マルチウエルプレートに移し、 一 80。Cに保存した。  Using a human liver cancer cell line, HB611 (Proc. Natl. Acad. Sci. USA 84, 444-448 (1987)), into which HBV residue was introduced, the method of Shuto et al. (Microbiol. Immunol., 40 (2), 153-159 (1996)), and the anti-V activity of the drug was measured. Briefly, ΗΒ611 cells were suspended in Dulbecco's modified MEM containing 10% bovine i¾fil, 1001 UZm1 benisiri G, 100 g / m1 streptomycin and lmg / m1 geneticin. 20,000 pieces per 1 well of a 96-well multiwell plate were applied. The cells were cultured in a carbon dioxide incubator at 37 ° C. for 3 days, and the cells were grown to confluent and then subjected to a test. The test drug was added to the cells, usually diluted with medium, in four steps of 10-fold serial dilution from a concentration of 100 / ig / inl, and cultured. The medium was replaced two to four days later with the medium containing the drug. Control cells were cultured in a drug-free medium. Seven days later, the culture supernatant was recovered from each well and transferred to another 96-well multiwell plate. Saved in C.
(2) 培養上清からの HBV DNAの精製 (2) Purification of HBV DNA from culture supernatant
の 96穴マルチウエルプレート （コ一二ング社製） の各ゥエルに等張リン 酸緩衝液 (P B S) で 1 0 (α gZm lに希釈した抗H B V表面抗原マゥス抗体Anti-HBV surface antigen mouse antibody diluted to 10 ( αgZml) with isotonic phosphate buffer (PBS) in each well of a 96-well multiwell plate (Korning)
(Zymed Laboratries製） 50 1を加え、 4°Cで約 1 6時間放置した。 各ゥェ ルの液層を捨て、 0. 1%の牛血清アルブミ ン （B SA) を含む P B S 100(Manufactured by Zymed Laboratries) 501 was added and left at 4 ° C for about 16 hours. Discard the liquid layer in each well, and add PBS 100 containing 0.1% bovine serum albumin (BSA).
1を加えて 37°Cで 2時間ィンキュベートした後、 使用直前まで 4 °Cで保存した。 各ゥエルを 0. 01%の Twe e n 20を含む P B S (P BS— 0. 0 l%Tw e e n 20) で 3回洗浄し、 0. 035%の Twe e n 20を含む PB Sを 10 1および回収保存しておいた培養上清 25 1を加え、 4でで一晚放置し、 培 養上清中の HBV粒子をプレートに吸着させた。 各ゥエルを PBS— 0. 01% Twe en20で 3回及び PBSで 2回洗浄した後、 0. 09NのNaOHと0. 01%の No n i d e t P— 40を含む溶液 25 n 1を加えて 37°Cで 1時間 インキュベートし、 HBV粒子から HBVの DNAを溶出させた。 0. 09Nの H C 1を含む 100 mMトリス一塩酸緩衝液 25 1を加えて中和し、 ポリメラ —ゼ連鎖反応 （PCR) に供した。 After adding 1 and incubating at 37 ° C for 2 hours, it was stored at 4 ° C until immediately before use. Wash each well three times with PBS containing 0.01% Tween 20 (PBS—0.0 l% Tween 20) to recover 10 1 PBS containing 0.035% Tween 20 Add the preserved culture supernatant 25 1 and leave it with 4 for 10 minutes. HBV particles in the culture supernatant were adsorbed to the plate. After washing each well three times with PBS-0.01% Tween20 and twice with PBS, add 25 n1 solution containing 0.09N NaOH and 0.01% Nonidet P-40 and add 37 ° C. After incubation at C for 1 hour, HBV DNA was eluted from the HBV particles. The mixture was neutralized by adding 25 1 of 100 mM Tris-hydrochloric acid buffer 251 containing 0.09N HC1, and subjected to a polymerase chain reaction (PCR).
(3) PCRでの増幅 (3) Amplification by PCR
PCRは市販のキット、 TaKaRa Ta q (宝酒造製） と、 Gene A mp sy s t em9600 (Perkin Elmer製） とを用いた。 プライマーは HB Vの S遺^?内の 372番から 483番が増幅されるように設定したものを用い た。 すなわち、 5' —末端にピオチンが結合した （+) 372— 401のプライ マ一 （5' —ビォチン一 TCGCTGGATGTGTCTGCGGCGTTTT AT) および （一） 460— 483のプライマー （5' — TAGAGGACAA ACGGGCAAC ATAC C) とを用いた。 抽出した H B Vの D N A溶液 5 1に P C R用混合溶液 45 β 1を加え （纖激： 10 mMトリス一塩酸緩衝液 (pH8. 3) 、 5 OmM KC 1、 1. 5mM MgC l2 、 0. 2mM o f ea ch dNTP、 6. 25 pmo 1 o f each プライマー、 1. 25U Amp 1 i Ta q DNA ポリメラーゼ） 、 P CRを行なった。 94 。C、 5分インキュベーションした後、 94で、 30秒一 55て、 15秒— 72°C、 1分を 30サイクル行ない、 最後に 72 °Cで 5分間反応させ、 4でで保存した。 For PCR, a commercially available kit, TaKaRa Taq (manufactured by Takara Shuzo) and Gene Amp system 9600 (manufactured by Perkin Elmer) were used. The primers used were designed so that Nos. 372 to 483 in the S residue of HBV were amplified. That is, the primer of (+) 372-401 (5'-biotin-TCGCTGGATGTGTCTGCGGCGTTTT AT) and the primer (1) of 460-483 (5'-TAGAGGACAA Using. To the extracted HBV DNA solution 51, add PCR mixed solution 45β1 (Fiber: 10 mM Tris-monohydrochloride buffer (pH 8.3), 5 OmM KC 1, 1.5 mM MgCl 2 , 0.2 mM of DNA dNTP, 6.25 pmo 1 of each primer, 1.25 U Amp 1 i Taq DNA polymerase) and PCR. 94. C. After 5 minutes incubation, 30 cycles of 94, 30 seconds and 55, 15 seconds—72 ° C., 1 minute were performed, and finally the reaction was performed at 72 ° C. for 5 minutes, and stored at 4.
_(4)ハイプリダイゼーシヨンおよび HBV DNAの検出 _ (4) Detection of hybridization and HBV DNA
平底の 96穴マルチウエルプレート （コ一二ング社製） の各ゥエルに 5 OmM クェン隨衝液 (pH9. 6) で lO^gZmlに希釈したストレプトアビジン の溶液を 75 β 1ずつ加え、 4°Cで約 16時間放置した。 各ゥエルの液層を捨て、 0. 1%B SAを含む P B S 1 00 // 1を加えて 37°Cで 2時間インキュベー シヨンした後、 使用直前まで 4°Cに保存した。 0. 05%Twe e n 20を含 む 0. 1 XS S PE溶液 （0. 1 XS S PE-0. 05% Twe e n 20) で 2回洗浄した後、 PCR産物 1 0 β 1を加えて室温で 1時間インキュベーショ ンし、 アビジンでコートされたプレートに、 5' 末端にビォチンを有する増幅さ れた DNAを吸着させた。 0. 1 Nの N a OHと 0. 15Mの N a C lを含む溶 液を 1 0 / 1加えて 5分間放置し、 吸着した DNAを変性させた。 Streptavidin diluted to lO ^ gZml with 5 OmM quenched buffer (pH 9.6) in each well of a flat-bottomed 96-well multiwell plate (manufactured by Koingen) Was added at 75 ° C. each, and the mixture was left at 4 ° C. for about 16 hours. The liquid layer in each well was discarded, PBS 100 // 1 containing 0.1% BSA was added, the mixture was incubated at 37 ° C for 2 hours, and stored at 4 ° C until immediately before use. After washing twice with 0.1 XS SPE solution containing 0.1% Tween 20 (0.1 XS SPE-0. 05% Tween 20), add PCR product 10β1 and room temperature After incubation for 1 hour, the amplified DNA having biotin at the 5 'end was adsorbed to the avidin-coated plate. A solution containing 0.1 N NaOH and 0.15 M NaCl was added at 10/1 and left for 5 minutes to denature the adsorbed DNA.
0. 1XS S PE-0. 05%Twe e n 20で 3回洗浄後、 ジゴキシゲニン で標識された l pmo 1のプローブ （ （一） 41 1— 437 : 5' — d i go x i g e n i n— AAGAAGATGAGGCATAGCAGCAGGATG) を 含む 40 1のハイプリダイゼイシヨン溶液 （4 X S S PE, 2. 5X Denh ardt' s solution , 100 mMトリスー塩酸緩衝液 （p H 7. 1 ) ， 0. 25m g/m 1 heat-denatured salmon sperm DNA， 0. 5 m g/ m 1 p o 1 y A) , 0. 05% Twe e n 20) を加え、 42°Cで 2時間インキュベーション した。 0. 1 XS S PE— 0. 05% Twe e n 20で 3回および PBSで 2回洗浄した後、 2000倍に希釈したアルカリフォスファターゼ結合抗ジォキ シゲニン抗体 (Boehringer Mannheim GmbH製） を 100 1加え、 室温で 1時間 インキュベーションした。 0. 1 XS S PE— 0. 05% Twe e n 20で 4回洗浄した後、 1 0 OmMの Na C 1と 5 OmMの Mg C 12 とを含む 100 mMトリス一塩酸緩衝液 (pH9. 5) で 1 m g/m 1に調整した 4一二トロフ ェニルフォスフヱイト溶液 1 00 1を加え、 37°Cで発色するまで 5から 15 分ィンキュベ一シヨンをした。 各ゥエルの 405 nmの吸光度をマイクロプレー トリーダーで測定して被験薬剤の抗 HBV活性を算出した。 薬剤を添加していな い対照の吸^ ¾を、 50%減少させる被験薬剤の濃度を、 50%有効濃度 （E C 50) とした。 After washing 3 times with 0.1XS SPE-0. 05% Tween 20, it contains the probe of lpmo1 labeled with digoxigenin ((1) 41 1—437: 5 '— di xixigenin—AAGAAGATGAGGCATAGCAGCAGAGGATG) 40 1 Hybridization solution (4 XSS PE, 2.5X Denhardt's solution, 100 mM Tris-HCl buffer (pH 7.1), 0.25 mg / m 1 heat-denatured salmon sperm DNA, 0 5 mg / m1po1yA) and 0.05% Tween 20) were added, and the mixture was incubated at 42 ° C for 2 hours. 0.1 XS SPE—After washing 3 times with 0.05% Tween 20 and 2 times with PBS, add 100 1 of 2000-fold diluted alkaline phosphatase-conjugated anti-dioxygenin antibody (manufactured by Boehringer Mannheim GmbH) and add room temperature. For 1 hour. After washing 4 times with 0. 1 XS S PE- 0. 05% Twe en 20, 1 0 OmM Na C 1 and 5 OmM Mg C 1 2 and 100 mM Tris monohydrochloride buffer containing of (pH 9. 5 ), Adjusted to 1 mg / m 1 with 1002 trophenylphosphite solution 1001, and incubated at 37 ° C for 5 to 15 minutes until color development. The absorbance at 405 nm of each well was measured with a microplate reader to calculate the anti-HBV activity of the test drug. The concentration of the test drug that reduces the absorption of the control to which no drug was added by 50% was increased by the 50% effective concentration (EC 50)
2)細胞毒性測定法： 2) Cytotoxicity measurement method:
被験薬剤の細胞毒性は MTT法で測定した。 培養上清を回収した後の HB 61 1細胞に新しい培地 100 1と 7. 5mg/m 1の MTT溶液 20 1とを加 えて炭酸ガスインキュベーター内で 37°C、 3時間培養した。 10% (v/v) Tr i t on X— 100を含む隱のイソプロパノール （イソプロパノール 5 00mlに対し、 濃塩酸を 2 m 1加えたもの） 溶液を 10 1加えてフオルマ ザンを溶出した。 完全にフオルマサンが溶解した後マイクロプレートリーダーで 540および 690 nmの吸光度を測定し、 被験薬剤の細胞毒性を算出した。 薬 剤を添加していない対照の吸光度を、 50%減少させる被験薬剤の を、 50 %細胞毒性 i« (cc50) とした。 The cytotoxicity of the test drug was measured by the MTT method. After the culture supernatant was recovered, HB61 1 cells were added with a fresh medium 100 1 and a 7.5 mg / m 1 MTT solution 201, and cultured at 37 ° C for 3 hours in a carbon dioxide gas incubator. Formalzan was eluted by adding 10 1 of a hidden isopropanol (500 ml of isopropanol to which 2 ml of concentrated hydrochloric acid was added) containing 10% (v / v) Triton X-100. After the formasan was completely dissolved, the absorbance at 540 and 690 nm was measured using a microplate reader to calculate the cytotoxicity of the test drug. The test drug that reduced the absorbance of the control to which no drug was added by 50% was defined as 50% cytotoxicity i «(cc 50 ).
3)試験結果： 3) Test results:
試験結果を下記表に示す The test results are shown in the table below
抗 HBV活性 Anti-HBV activity
化 合 物 細 胞 毒 性  Compound cell toxic
E C5o： β g/m 1 C C50： gZm 1 -L-4' —チオアラビノフラノシルシトシン 15. 3 >100 a-L-4' ーチオアラビノフラノシルチミン 6. 80 > 100 EC 5 o: β g / m 1 CC 50: gZm 1 -L-4 '- thio-arabinofuranosyl cytosine 15. 3> 100 aL-4' over thio-arabinofuranosyl thymine 6.80> 100
2, 6—ジァミノ一 （a— L— 4' —チオアラビノ 19. 8 >100 フラノシル） プリン  2, 6-diamino (a—L—4'-thioarabino 19.8> 100 furanosyl) pudding
2, 6—ジアミノー （；8— L— 4' —チオアラビノ 4. 37 > 100 フラノシル） プリン a-L-4' —チオアラビノフラノシルアデニン 5. 15 > 100 2,6-diamino-(; 8-L-4'-thioarabino 4.37> 100 furanosyl) purine aL-4'-thioarabinofuranosyl adenine 5.15> 100
製剤例 1 ：翻 Formulation Example 1
本発明化合物 30. Omg  Compound of the present invention 30.Omg
微粉末セルロース 25. Omg  Fine powdered cellulose 25.Omg
乳糖 39. 5mg  Lactose 39.5 mg
スターチ 40. Omg  Starch 40. Omg
タルク 5. Omg  Talc 5. Omg
ステアリン酸マグネシム 0. 5 m g  Magnesium stearate 0.5 mg
上言 H ^から常法によつて錠剤を調製する t 製剤例 2：カプセル剤 Preparation of tablets from H ^ in a conventional manner t Formulation example 2: Capsules
本発明化合物 30. Omg  Compound of the present invention 30.Omg
40. Omg  40.Omg
スターチ 15. Omg  Starch 15. Omg
タルク 5. Omg  Talc 5. Omg
上言 Η β¾から常法によつて力プセル剤を調製する。 製剤例 3：膝剤  From the above {β}, prepare a forcepsel in a conventional manner. Formulation Example 3: Knee
本発明化合物 30. Omg  Compound of the present invention 30.Omg
グルコース 100. Omg  Glucose 100. Omg
上記 β¾を注射用精製水に溶解して' ^ί剤を調製する ( 産 H±の利用可纖  Dissolve the above β¾ in purified water for injection to prepare a ί ^ ί agent.
前述の試験例の結果から明らかなように、本発明の L— 4' —チオアラビノフ ラノヌクレオシド化合物は毒性がなく、 B型肝炎ウィルス (HBV) に対して顕 著な抑制効果を有すことから抗肝炎ウィルス ^^物、特に、抗 B型肝炎ウィルス 剤として有用であり、 医薬品としての開発が期待されうるものである。 As is clear from the results of the above-mentioned test examples, the L-4′-thioarabinofuranonucleoside compound of the present invention has no toxicity and has a remarkable inhibitory effect on hepatitis B virus (HBV). Hepatitis virus ^^, especially anti-hepatitis B virus It is useful as an agent and can be expected to be developed as a pharmaceutical.
また、 前述した L— 4 ' —チオアラビノフラノヌクレオシド化合物の合成法は、 天然糖の D—キシロースから簡便な方法で目的ィ匕合物を導くことができ、 極めて 実用的な方法である。  In addition, the method for synthesizing the above-mentioned L-4′-thioarabinofuranonucleoside compound is an extremely practical method that can derive the desired compound from the natural sugar D-xylose by a simple method.

Claims

請求の 範囲 The scope of the claims
1. 下式 [I] で表される L一 4' ーチオアラビノフラノヌクレオシド化合 物： 1. L-1 4'-thioarabinofuranonucleoside compound represented by the following formula [I]:
Figure imgf000032_0001
Figure imgf000032_0001
(式中、 Bはピリミジン、 プリン、 ァザプリンおよびデァザプリンからな 群よ り選択される核酸塩基を示し、 それらはハロゲン原子、 アルキル基、 ハロアルキ ル基、 アルケニル基、 ハロアルケニル基、 アルキニル基、 アミノ基、 アルキルァ ミノ基、水酸基、 ヒドロキシァミノ基、 アミノキシ基、 アルコキシ基、 メルカプ ト基、 アルキルメルカプト基、 ァリール基、 ァリールォキシ基またはシァノ基に よって置換されていてもよい。 ） (In the formula, B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine and azapurine, which is a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkynyl group, an amino group. , An alkylamino group, a hydroxyl group, a hydroxyamino group, an aminoxy group, an alkoxy group, a mercapto group, an alkylmercapto group, an aryl group, an aryloxy group or a cyano group.)
2. a-L-4' ーチオアラビノフラノピリ ミジンヌクレオシドである、 請 求項 1記載の L— 4 ' —チオアラビノフラノヌクレオシド化合物。  2. The L-4′-thioarabinofurano nucleoside compound according to claim 1, which is an a-L-4′-thioarabinofuranopyrimidine nucleoside.
3. α— L— 4' ーチオアラビノフラノプリンヌクレオシドである、 請求項 1記載の L— 4' ーチオアラビノフラノヌクレオシド化合物。  3. The L-4′-thioarabinofurano nucleoside compound according to claim 1, which is an α-L-4′-thioarabinofuranopurine nucleoside.
4. /S-L-4' —チオアラビノフラノプリンヌクレオシドである、 請求項 1記載の L— 4' —チオアラビノフラノヌクレオシド化合物。  4. The L-4′-thioarabinofurano nucleoside compound according to claim 1, which is a / S-L-4'-thioarabinofuranopurine nucleoside.
5. 下式 [I] で表される L一 4' —チオアラビノフラノヌクレオシド化合 物を活¾ ^分として含有する医薬糸！ ^物： 5. Pharmaceutical thread containing as an active ingredient L-1 4'-thioarabinofuranonucleoside compound represented by the following formula [I]! ^ Things:
Figure imgf000033_0001
Figure imgf000033_0001
(式中、 Bはピリミジン、 プリン、 ァザプリンおよびデァザプリンからなる群よ り選択される核酸塩基を示し、 それらはハロゲン原子、 アルキル基、 ハロアルキ ル基、 アルケニル基、 ハロアルケニル基、 アルキニル基、 アミノ基、 アルキルァ ミノ基、 水酸基、 ヒドロキシァミノ基、 アミノキシ基、 アルコキシ基、 メルカプ ト基、 アルキルメルカプト基、 ァリ一ル基、 ァリールォキシ基またはシァノ基に よって置換されていてもよい。 ） (In the formula, B represents a nucleobase selected from the group consisting of pyrimidine, purine, azapurine, and azapurine, and is a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkynyl group, an amino group. May be substituted by an alkylamino group, a hydroxyl group, a hydroxyamino group, an aminoxy group, an alkoxy group, a mercapto group, an alkylmercapto group, an aryl group, an aryloxy group or a cyano group.)
6. L一 4 ' ーチオアラビノフラノヌクレオシド化合物が、 a - L - 4' 一 チオアラビノフラノプリンヌクレオシドまたは yS— L— 4' —チオアラビノフラ ノプリンヌクレオシドから選ばれる、 請求項 5記載の 物。  6. The product according to claim 5, wherein the L-14′-thioarabinofuranonucleoside compound is selected from a-L-4′-thioarabinofuranopurine nucleoside or yS—L—4′-thioarabinofuranopurine nucleoside. .
7. L— 4 ' ーチオアラビノフラノヌクレオシド化合物が、 一 L一 4' 一 チオアラビノフラノピリミジンヌクレオシドである、 請求項 5記載の組成物。  7. The composition according to claim 5, wherein the L-4′-thioarabinofurano nucleoside compound is 1 L-1 4′-thioarabinofuranopyrimidine nucleoside.
8. 抗肝炎ウィルス組成物である、 請求項 5記載の組成物。  8. The composition according to claim 5, which is an anti-hepatitis virus composition.
9. 抗 B型肝炎ウィルス剤である、 請求項 5記載の組成物。  9. The composition according to claim 5, which is an anti-hepatitis B virus agent.
1 0. 下記の 4つの工程を含んでなる L— 4 ' ーチオアラビノフラノヌクレ オシド化合物の合成法：  10. A method for synthesizing an L-4′-thioarabinofuranonucleoside compound comprising the following four steps:
第 1工程： First step:
D—キシロースを、 アルコール溶媒中、 酸の存在下で 1位をアルキルィ匕した後、 水酸基を保護し、 次いで 1位アルキル基の酸,による加水分解、 引き続き還元 剤による処理により式 [I I] で表される化合物を得る工程： After alkylating D-xylose at the 1st position in the presence of an acid in an alcohol solvent, Step of protecting the hydroxyl group, then hydrolyzing the 1-position alkyl group with an acid, and subsequently treating with a reducing agent to obtain a compound represented by the formula [II]:
D― xyloseD- xylose
Figure imgf000034_0001
Figure imgf000034_0001
(式中、 1^ はアルキル基を示す） 第 2工程： (Where 1 ^ represents an alkyl group) Second step:
式 [I Π で表される化合物の水酸基をメシル化剤によりメシルイ匕またはトシ ル化剤により トシル化した後、 硫ィ匕ナトリウムで処理して式 [I I I] で表され る化合物を得る工程：  A step of obtaining a compound represented by the formula [III] by treating a hydroxyl group of the compound represented by the formula [I] with a mesylating agent or by tosylating the compound with a tosylating agent, followed by treating with sodium sulfate.
Figure imgf000034_0002
Figure imgf000034_0002
[HI]  [HI]
(式中、 R1 は前記と同義） 第 3工程： (Wherein, R 1 is as defined above) Third step:
式 [I I I] で表される化合物を適当な酸化剤によりスルホキシド体に導き. 更に酸無水物で処理することでプンメラ一 （Pumme r e r) 転移を行ない- 式 [I V] で表される化合物を得る工程：  The compound represented by the formula [III] is converted into a sulfoxide form with an appropriate oxidizing agent. The compound is further treated with an acid anhydride to perform a Pumme rer transition to obtain a compound represented by the formula [IV]. Process:
Figure imgf000035_0001
Figure imgf000035_0001
(式中、 1^ は、 前記と同義； Rり はァシル基を示す） 第 4工程： (Where 1 ^ is as defined above; R is an acyl group). Fourth step:
式 [I V] で表される化合物にグリコシル化反応により核酸塩基を導入し、 更 に糖部保護基を脱保護して式 [I] で表される化合物を得る工程：  A step of introducing a nucleobase to the compound represented by the formula [IV] by a glycosylation reaction, and further deprotecting the sugar protecting group to obtain a compound represented by the formula [I]:
Figure imgf000035_0002
Figure imgf000035_0002
[I] [I]
(式中、 Bはピリ ミジン、 プリン、 ァザプリンおよびデァザプリンからなる群よ り選択される核酸塩基を示し、 それらはハロゲン原子、 アルキル基、 ハロアルキ ル基、 アルケニル基、 ハロアルケニル基、 アルキニル基、 アミノ基、 アルキルァ ミノ基、 水 ?S、 ヒドロキシァミノ基、 アミノキシ基、 アルコキシ基、 メルカプ ト基、 アルキルメルカプト基、 ァリール基、 ァリールォキシ基またはシァノ基に よって置換されていてもよい。 ） (Where B is a group consisting of pyrimidine, pudding, azapurine and dazapurine Selected from the group consisting of a halogen atom, an alkyl group, a haloalkyl group, an alkenyl group, a haloalkenyl group, an alkynyl group, an amino group, an alkylamino group, water-S, a hydroxyamino group, an aminoxy group, It may be substituted by an alkoxy group, a mercapto group, an alkylmercapto group, an aryl group, an aryloxy group or a cyano group. )
PCT/JP1999/000827 1998-02-25 1999-02-24 L-4'-arabinofuranonucleoside compound and medicine composition comprising the same WO1999043690A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP10/43893 1998-02-25
JP4389398 1998-02-25

Publications (1)

Publication Number Publication Date
WO1999043690A1 true WO1999043690A1 (en) 1999-09-02

Family

ID=12676398

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1999/000827 WO1999043690A1 (en) 1998-02-25 1999-02-24 L-4'-arabinofuranonucleoside compound and medicine composition comprising the same

Country Status (1)

Country Link
WO (1) WO1999043690A1 (en)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6576621B1 (en) 1998-07-23 2003-06-10 Southern Research Institute Preparation of thioarabinofuranosyl compounds and use thereof
WO2015125782A1 (en) * 2014-02-18 2015-08-27 富士フイルム株式会社 Method for producing thiolane-skeleton glycoconjugate, and thiolane-skeleton glycoconjugate
AU2013203974B2 (en) * 2000-05-23 2016-06-02 Idenix Pharmaceuticals, Inc. Methods and compositions for treating hepatitis C virus
JP2016185979A (en) * 2012-08-13 2016-10-27 富士フイルム株式会社 SYNTHETIC INTERMEDIATE OF 1-(2-DEOXY-2-FLUORO-4-THIO-β-D-ARABINOFURANOSYL)CYTOSINE, SYNTHETIC INTERMEDIATE OF THIONUCLEOSIDE, AND METHOD FOR PRODUCING THEM
US9815812B2 (en) 2014-02-19 2017-11-14 Fujifilm Corporation Thiopyranose compound and method for producing same
US9884882B2 (en) 2014-02-18 2018-02-06 Fujifilm Corporation Method for producing thiolane skeleton-type glycoconjugate, and thiolane skeleton-type glycoconjugate
US9896471B2 (en) 2012-03-28 2018-02-20 Fujifilm Corporation Salt of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine
US9968628B2 (en) 2000-05-26 2018-05-15 Idenix Pharmaceuticals Llc Methods and compositions for treating flaviviruses and pestiviruses
US10059734B2 (en) 2014-10-31 2018-08-28 Fujifilm Corporation Thionucleoside derivative or salt thereof, and pharmaceutical composition
US11141421B2 (en) 2018-01-29 2021-10-12 Fujifilm Corporation Antitumor agent for biliary tract cancer and method for treating biliary tract cancer
US11369625B2 (en) 2016-08-31 2022-06-28 Fujifilm Corporation Anti-tumor agent, anti-tumor effect enhancer, and anti-tumor kit

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05505791A (en) * 1989-10-04 1993-08-26 ユニバーシティ　オブ　バーミンガム More antiviral pyrimidine nucleosides
JPH0925289A (en) * 1995-07-14 1997-01-28 Yamasa Shoyu Co Ltd Production of 4'-thioarabinopyrimidine nucleoside
JPH09249690A (en) * 1995-07-14 1997-09-22 Yamasa Shoyu Co Ltd 4'-thioarabninopurine nucleoside

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH05505791A (en) * 1989-10-04 1993-08-26 ユニバーシティ　オブ　バーミンガム More antiviral pyrimidine nucleosides
JPH0925289A (en) * 1995-07-14 1997-01-28 Yamasa Shoyu Co Ltd Production of 4'-thioarabinopyrimidine nucleoside
JPH09249690A (en) * 1995-07-14 1997-09-22 Yamasa Shoyu Co Ltd 4'-thioarabninopurine nucleoside

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
JOHN A. SECRIST III et al., "Synthesis and Biological Activity of Certain 4'-Thio-D-Arabinofuranosylpurine Nucleosides", JOURNAL OF MEDICINAL CHEMISTRY, Vol. 41, No. 20, (24 September 1998), pages 3865-3871. *
JOHN A. SECRIST III et al., "The Synthesis and Biological Activities of Certain 4'-Thionucleosides", NUCLEOSIDES & NUCLEOTIDES, Vol. 14, No. 3-5, (1995), pages 675-686. *
MACHIDA H. et al., "Anti-Herpesvirus Activity Profile of 4'-Thioarabinofuranosyl Purine and Uracil Nucleosides and Activity of 1-beta-D-2'-Fluoro-4'-Thioarabinofuranosyl Guanine and 2,6-Diaminopurine Against Clinical Isolates of Human Cytomegalovirus", ANTIVIRAL RESEARCH, Vol. 39, No. 2, (August 1998), pages 129-137. *
OTOTANI N. et al., "Preparation and Antitumor Activity of 4'-Thio Analogs of 2,2'-Anhydro-1-beta-D-Arabinofuranosylcytos ine", JOURNAL OF MEDICINAL CHEMISTRY, Vol. 17, No. 5, (1974), pages 535-537. *
SATOH H. et al., "Synthesis of L-Enantiomers of 4'-Thioarabinofuranosyl Pyrimidine Nucleosides", BIOORGANIC AND MEDICINAL CHEMISTRY LETTERS, Vol. 8, No. 5, (May 1998), pages 989-992. *
SIDDIQI S.M. et al., "Search for New Purine- and Ribose-Modified Adenosine Analogues as Selective Agonists and Antagonists at Adenosine Receptors", JOURNAL OF MEDICINAL CHEMISTRY, Vol. 38, No. 7, (1995), pages 1174-1188. *
WHISTLER R.L. et al., "4-Thio-D-Arabinofuranosylpyrimidine Nucleosides", J. ORG. CHEM., Vol. 36, No. 1, (1971), pages 108-110. *
YOSHIMURA Y. et al., "A Facile, Alternative Synthesis of 4'-Thioarabinonucleosides and Their Biological Activities", JOURNAL OF MEDICINAL CHEMISTRY, Vol. 40, No. 14, (1997), pages 2177-2183. *
YOSHIMURA Y. et al., "Synthesis and Biological Activities of 2'-Modified 4'-Thionucleosides", NUCLEOSIDES & NUCLEOTIDES, Vol. 16, No. 7-9, (1997), pages 1103-1106. *

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6870052B2 (en) 1998-07-23 2005-03-22 Southern Research Institute Preparation of thioarabinofuranosyl compounds and use thereof
US7138385B2 (en) 1998-07-23 2006-11-21 Southern Research Institute Thioarabinofuranosyl compounds and use thereof
US7691820B2 (en) 1998-07-23 2010-04-06 Southern Research Institute Preparation of thioarabinofuranosyl compounds and use thereof
US8178510B2 (en) 1998-07-23 2012-05-15 Southern Research Institute Preparation of thioarabinofuranosyl compounds and use thereof
US6576621B1 (en) 1998-07-23 2003-06-10 Southern Research Institute Preparation of thioarabinofuranosyl compounds and use thereof
AU2013203974B2 (en) * 2000-05-23 2016-06-02 Idenix Pharmaceuticals, Inc. Methods and compositions for treating hepatitis C virus
US9968628B2 (en) 2000-05-26 2018-05-15 Idenix Pharmaceuticals Llc Methods and compositions for treating flaviviruses and pestiviruses
US9896471B2 (en) 2012-03-28 2018-02-20 Fujifilm Corporation Salt of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine
JP2016185979A (en) * 2012-08-13 2016-10-27 富士フイルム株式会社 SYNTHETIC INTERMEDIATE OF 1-(2-DEOXY-2-FLUORO-4-THIO-β-D-ARABINOFURANOSYL)CYTOSINE, SYNTHETIC INTERMEDIATE OF THIONUCLEOSIDE, AND METHOD FOR PRODUCING THEM
US10093645B2 (en) 2012-08-13 2018-10-09 Fujifilm Corporation Synthetic intermediate of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine, synthetic intermediate of thionucleoside, and method for producing the same
US10570112B2 (en) 2012-08-13 2020-02-25 Fujifilm Corporation Synthetic intermediate of 1-(2-deoxy-2-fluoro-4-thio-β-D-arabinofuranosyl)cytosine, synthetic intermediate of thionucleoside, and method for producing the same
JP2015172033A (en) * 2014-02-18 2015-10-01 富士フイルム株式会社 Method for producing thiolane skeleton-type glycoconjugate, and thiolane skeleton-type glycoconjugate
US9884882B2 (en) 2014-02-18 2018-02-06 Fujifilm Corporation Method for producing thiolane skeleton-type glycoconjugate, and thiolane skeleton-type glycoconjugate
WO2015125782A1 (en) * 2014-02-18 2015-08-27 富士フイルム株式会社 Method for producing thiolane-skeleton glycoconjugate, and thiolane-skeleton glycoconjugate
US9815812B2 (en) 2014-02-19 2017-11-14 Fujifilm Corporation Thiopyranose compound and method for producing same
US10059734B2 (en) 2014-10-31 2018-08-28 Fujifilm Corporation Thionucleoside derivative or salt thereof, and pharmaceutical composition
US10385089B2 (en) 2014-10-31 2019-08-20 Fujifilm Corporation Thionucleoside derivative or salt thereof, and pharmaceutical composition
US11369625B2 (en) 2016-08-31 2022-06-28 Fujifilm Corporation Anti-tumor agent, anti-tumor effect enhancer, and anti-tumor kit
US11141421B2 (en) 2018-01-29 2021-10-12 Fujifilm Corporation Antitumor agent for biliary tract cancer and method for treating biliary tract cancer

Similar Documents

Publication Publication Date Title
EP1177202B1 (en) 4&#39;-c-ethynyl purine nucleosides
US6833361B2 (en) Nucleosides having bicyclic sugar moiety
JP4253342B2 (en) 4&#39;-C-substituted-2-haloadenosine derivatives
US5607922A (en) 1,5-anhydrohexitol nucleoside analogues
EP1572705A2 (en) Sugar modified nucleosides as viral replication inhibitors
HU199870B (en) Process for producing purine arabinoside derivatives with antiviral effect and pharmaceutical compositions comprising same
CZ120699A3 (en) Monocyclic l-nucleoside analogs and their utilization
EP1745573A2 (en) METHODS OF MANUFACTURE OF 2 -DEOXY-&amp;bgr;-L-NUCLEOSIDES
JP2008069182A (en) 4&#39;-c-substituted-2-haloadenosine derivative
WO1999043690A1 (en) L-4&#39;-arabinofuranonucleoside compound and medicine composition comprising the same
WO1997037993A1 (en) 9-(2-DEOXY-2-FLUORO-4-THIO-β-D-ARABINOFURANOSYL)PURINE DERIVATIVES
JP4430307B2 (en) Method for producing 2&#39;-halo-β-L-arabinofuranosyl nucleoside
JPH06228186A (en) 2&#39;-deoxy-@(3754/24)2&#39;s)-alkylpyrimidine nucleoside derivative
JP4076114B2 (en) 4&#39;-C-ethynylpurine nucleoside compounds
JPH0853490A (en) 2&#39;-deoxy-2&#39;,2&#39;-dihalogeno-4&#39;-thionucleoside
US5420115A (en) Method for the treatment of protoza infections with 21 -deoxy-21 -fluoropurine nucleosides
WO1992021343A1 (en) Novel pyrazinone n-oxide nucleosides and analogs thereof
US7595390B2 (en) Industrially scalable nucleoside synthesis
JP4039790B2 (en) 4&#39;-C-ethynylpyrimidine nucleoside compounds
JPWO2003068796A1 (en) 4&#39;-C-cyano-2&#39;-deoxypurine nucleoside
JPH1087687A (en) 5-substituted-1-(2-deoxy-2-fluoro-4-thio-beta-d-arabinofuranosyl)uracil
JP2008110983A (en) 4&#39;-c-ethynyl nucleoside compound
JPH10168096A (en) 2&#39;-azido-2&#39;-deoxy-4&#39;-thioribonucleoside

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA CN JP KR US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: KR

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

Ref document number: 2000533442

Format of ref document f/p: F