HK1235794A1

HK1235794A1 - Thionucleoside derivative or salt thereof, and pharmaceutical composition

Info

Publication number: HK1235794A1
Application number: HK17109674.5A
Authority: HK
Inventors: 国吉荣喜; 中川大辅; 松本拓也; 吉光佑二
Original assignee: 富士胶片株式会社
Priority date: 2014-10-31
Filing date: 2015-11-02
Publication date: 2018-03-16

Description

Thionucleoside derivative or salt thereof and pharmaceutical composition

Technical Field

The present invention relates to a thionucleoside derivative or a salt thereof, which is useful as a therapeutic agent for tumors, and a pharmaceutical composition.

Background

It is known that 1- (2-deoxy-2-fluoro-4-thio- β -D-arabinofuranosyl) cytosine (hereinafter, also referred to as compound a) which is a thionucleoside has an excellent antitumor effect and is useful for the treatment of malignant tumors (for example, lung cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, kidney cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma) and the like (patent document 1).

On the other hand, gemcitabine (2 ', 2 ' -difluoro-2 ' -deoxycytidine), which is a nucleoside, is known to be effective against cancerous tumors such as large intestine, lung, pancreas, breast, bladder, ovary and the like (non-patent documents 1 and 2). Gemcitabine has become a standard chemotherapy for pancreatic cancer after FDA approval in 1996 and has recently been approved for non-small cell lung cancer, ovarian cancer, bladder cancer and breast cancer.

Of these, innate or acquired resistance to nucleoside analogs (e.g., gemcitabine, 5-fluorouracil, cytarabine, and fludarabine) is a common problem in cancer therapy and is considered to be an important cause of low patient survival. Gemcitabine also faces many problems of innate or acquired drug resistance as with other nucleoside analogues (non-patent document 3). It has been reported that tumor cells reduce the overall survival of cancer patients by acquiring resistance to gemcitabine (non-patent document 4).

In order to overcome resistance to gemcitabine, development of new drugs based on resistance mechanisms has been carried out, and for example, compounds obtained by adding an amide phosphite to gemcitabine are known (non-patent document 3).

Documents of the prior art

Patent document

Patent document 1: international publication No. 1997/038001 pamphlet

Non-patent document

Non-patent document 1: cancer Research, volume 50, pages 4417 to 4422, 1990

Non-patent document 2: Anti-Cancer Drugs, volume 6, pages 7-13, 1995

Non-patent document 3: journal of Medicinal Chemistry, volume 57, pages 1531 to 1542, 2014

Non-patent document 4: neoplasia, volume 12, pages 807-817, year 2010

Disclosure of Invention

Problems to be solved by the invention

Disclosed are a compound and a pharmaceutical composition which exhibit excellent drug efficacy against tumors, particularly tumors that have acquired gemcitabine resistance.

Means for solving the problems

The present inventors have conducted intensive studies and, as a result, have found that a specific derivative of compound a exhibits excellent drug efficacy against tumors, particularly tumors that have acquired gemcitabine resistance, and have completed the present invention.

The present invention provides the following aspects.

(1) A thionucleoside derivative represented by the general formula [1] or a salt thereof:

[ chemical formula 1]

In the formula, R¹Represents an optionally protected hydroxyl group, an optionally substituted C_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted; r²Represents optionally substituted C_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted; or R¹And R²Or a 5-to 10-membered nitrogen-phosphorus-containing heterocycle which may be substituted, a 5-to 10-membered oxygen-phosphorus-containing heterocycle which may be substituted, or a 5-to 10-membered nitrogen-oxygen-phosphorus-containing heterocycle; r³Represents a hydrogen atom; or R²And R³Or with R²Bonded phosphorus atom, oxygen atom, methylene group, 2 carbon atoms constituting tetrahydrothiophene ring and R³The bonded oxygen atoms form together a 6-to 10-membered oxygen-phosphorus-containing heterocycle which may be substituted.

(2) The thionucleoside derivative or salt thereof according to (1), wherein R¹A hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a.

(3) The thionucleoside derivative or salt thereof according to (1) or (2), wherein R²Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a.

(4) The thionucleoside derivative or salt thereof according to any one of (1) to (3), wherein R¹A hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a; r²Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a.

(5) The thionucleoside derivative or salt thereof according to any one of (1) to (4), wherein R¹Is aryloxy which may be substituted by 1 or more substituents selected from substituent group A, R²Is an amino group which may be substituted by 1 or more substituents selected from substituent group A, or,

R¹Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6Alkoxy radical, R²Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6An alkoxy group.

(6) The thionucleoside derivative or a salt thereof according to (1), wherein the thionucleoside derivative is selected from the group consisting of

(2S) -benzyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate, and pharmaceutically acceptable salts thereof,

(2S) -methyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (4-chlorophenoxy) phosphoryl) amino) propanoate, and,

(2S) -cyclobutyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate, and salts thereof,

(2S) -methyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (4-bromophenoxy) phosphoryl) amino) propionate, pharmaceutically acceptable salts thereof, and pharmaceutically acceptable salts thereof,

(2S) -methyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (4-iodophenoxy) phosphoryl) amino) propionate, pharmaceutically acceptable salts thereof, and pharmaceutically acceptable salts thereof,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (S-pivaloyl-2-mercaptoethane-1-yl) phosphate,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (S-isobutyryl-2-mercaptoethane-1-yl) phosphate,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (S-propionyl-2-mercaptoethane-1-yl) phosphate,

((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (2- (pivaloylthio) ethoxy) phosphoryl) oxy) methyl pivalate,

S- (2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (((isopropoxycarbonyl) oxy) methoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate, and mixtures thereof,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (pivaloyloxymethyl) phosphate,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methylbis (2- ((2- (benzyloxy) ethyl) disulfide) ethyl) phosphate,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methylbis (2- ((2-pivaloyloxyethyl) disulfide) ethyl) phosphate,

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methylbis (2- ((2-isobutyryloxyethyl) disulfide) ethyl) phosphate,

S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate, and mixtures thereof,

4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- ((((RS) -2-oxide-4H-benzo [ d ] [1, 3, 2] dioxaphosphino-2-yl) oxy) methyl) tetrahydrothien-2-yl) pyrimidin-2 (1H) -one,

(2S) -benzyl 2- (((2RS, 4aR, 6R, 7S, 7aS) -6- (4-amino-2-oxopyrimidin-1 (2H) -yl) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-2-yl) amino) propionate, and

(2S) -isopropyl 2- (((2RS, 4aR, 6R, 7S, 7aS) -6- (4-amino-2-oxopyrimidin-1 (2H) -yl) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-2-yl) amino) propanoate.

(7) A pharmaceutical composition comprising the thionucleoside derivative or salt thereof according to any one of (1) to (6).

(8) The pharmaceutical composition according to (7), which is used for the treatment of tumors.

(9) The pharmaceutical composition of (7), which is used for the treatment of lung cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, renal cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma, or malignant lymphoma.

The invention also provides the following scheme.

(a) A thionucleoside derivative represented by the general formula [1] defined above or a salt thereof for use as a medicine.

(b) A thionucleoside derivative represented by the general formula [1] or a salt thereof, which is useful for the treatment of tumors.

(c) A thionucleoside derivative represented by the general formula [1] or a salt thereof for use in the treatment of lung cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, renal cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma or malignant lymphoma.

(d) A pharmaceutical composition comprising a pharmacologically acceptable additive together with a thionucleoside derivative represented by the general formula [1] or a salt thereof.

(e) Use of a thionucleoside derivative represented by the general formula [1] or a salt thereof for treating tumors in the production of a medicine.

(f) Use of a thionucleoside derivative represented by the general formula [1] or a salt thereof for the production of a medicament for the treatment of lung cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, renal cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma or malignant lymphoma.

(g) A method for treating a tumor, which comprises the step of administering a therapeutically effective amount of a thionucleoside derivative represented by the general formula [1] or a salt thereof to a subject (a mammal including a human) requiring such treatment.

(h) A method for treating lung cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, kidney cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma or malignant lymphoma, which comprises administering a therapeutically effective amount of a thionucleoside derivative represented by the general formula [1] or a salt thereof to a subject (a mammal including a human) in need of such treatment.

Effects of the invention

The thionucleoside derivative or salt thereof of the present invention is useful as a therapeutic agent for tumors. The thionucleoside derivative or a salt thereof of the present invention is useful particularly as a therapeutic agent for tumors that have acquired gemcitabine resistance.

Detailed Description

The present invention will be described in detail below.

In the present invention, unless otherwise specified, the terms have the following meanings.

The halogen atom means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

C_1-6The alkyl group means a straight-chain or branched C group such as methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, 2-pentyl, 3-pentyl and hexyl group_1-6An alkyl group.

C_1-20Alkyl means methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, 2-methylbutyl, 2-pentyl, 3-pentyl, hexyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, heptyl, 1-methylhexyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 1-propylbutyl, 4-dimethylpentyl, octyl, 1-methylheptyl, heptyl, pentyl, hexyl, heptyl, hexyl, 2-methylbutyl, 2-methylheptyl, 3-methylheptyl, 4-methylheptyl, 5-methylheptyl, 6-methylheptyl, 1-propylpentyl, 2-ethylhexyl, 5-dimethylhexyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecylStraight or branched C such as octadecyl, nonadecyl and eicosyl_1-20An alkyl group.

C_1-6Alkylsulfonyl means C such as methylsulfonyl, ethylsulfonyl and propylsulfonyl_1-6An alkylsulfonyl group.

C_1-6The alkylsulfonyloxy group is a methylsulfonyloxy group, an ethylsulfonyloxy group or the like C_1-6An alkylsulfonyloxy group.

C_1-6The alkyldithio group means a straight-chain or branched C group such as methyldithio, ethyldithio, propyldithio, isopropyldithio, butyldithio, sec-butyldithio, isobutyldithio, tert-butyldithio, pentyldithio, isopentyldithio, 2-methylbutyldithio, 2-pentyldithio, 3-pentyldithio and hexyldithio_1-6An alkyl disulfide group.

C_2-6Alkenyl refers to straight or branched chain C such as vinyl, allyl, propenyl, isopropenyl, butenyl, isobutenyl, 1, 3-butadienyl, pentenyl and hexenyl_2-6An alkenyl group.

C_2-6Alkynyl refers to straight-chain or branched-chain C such as ethynyl, propynyl, butynyl, pentynyl and hexynyl_2-6Alkynyl.

C_3-8Cycloalkyl is C such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl_3-8A cycloalkyl group.

C_3-8Cycloalkyl-dithio is C such as cyclopropyl-dithio, cyclobutyl-dithio, cyclopentyl-dithio, cyclohexyl-dithio and cycloheptyl-dithio_3-8Cycloalkyl disulfide group.

C_1-6Alkoxy is straight or branched chain C such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, etc_1-6An alkoxy group.

C_1-20The alkoxy group means a straight-chain or branched C-alkoxy group such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, nonyloxy, decyloxy, undecyloxy, dodecyloxy, tridecyloxy, tetradecyloxy, pentadecyloxy, hexadecyloxy, heptadecyloxy, octadecyloxy, nonadecyloxy and eicosyloxy_1-20An alkoxy group.

C_1-6Alkoxy radical C_1-6The alkyl group is a methoxymethyl group, a 1-ethoxyethyl group or the like C_1-6Alkoxy radical C_1-6An alkyl group.

C_1-6The alkoxycarbonyl group means a straight-chain or branched C-chain carbonyl group such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl and the like_1-6An alkoxycarbonyl group.

C_1-20The alkoxycarbonyl group means a straight-chain or branched C-chain alkoxycarbonyl group such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, sec-butoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, heptyloxycarbonyl, octyloxycarbonyl, nonyloxycarbonyl, decyloxycarbonyl, undecyloxycarbonyl, dodecyloxycarbonyl, tridecyloxycarbonyl, tetradecyloxycarbonyl, pentadecyloxycarbonyl, hexadecyloxycarbonyl, heptadecyloxycarbonyl, octadecyloxycarbonyl, nonadecyloxycarbonyl and icosyloxycarbonyl_1-20An alkoxycarbonyl group.

C_1-6The alkoxycarbonyloxy group is a straight-chain or branched C-chain group such as methoxycarbonyloxy, ethoxycarbonyloxy, propoxycarbonyloxy, isopropoxycarbonyloxy, butoxycarbonyloxy, isobutoxycarbonyloxy, sec-butoxycarbonyloxy, tert-butoxycarbonyloxy, pentyloxycarbonyloxy and hexyloxycarbonyloxy_1-6An alkoxycarbonyloxy group.

C_3-8Cycloalkoxy is C such as cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy and cycloheptyloxy_3-8A cycloalkoxy group.

C_3-8Cycloalkyloxycarbonyl means C such as cyclopropyloxycarbonyl, cyclobutyloxycarbonyl, cyclopentyloxycarbonyl, cyclohexyloxycarbonyl, cycloheptyloxycarbonyl and cyclooctyloxycarbonyl_3-8A cycloalkoxycarbonyl group.

Aryl means phenyl or naphthyl.

Aryloxy means phenoxy, naphthalene-1-oxy or naphthalene-2-oxy.

Arylsulfonyl means phenylsulfonyl, p-toluenesulfonyl, or naphthylsulfonyl.

Arylsulfonyloxy means phenylsulfonyloxy or p-tolylsulfonyloxy.

Aryl-disulfide means phenyl-disulfide or naphthyl-disulfide.

Aromatic C_1-6The alkyl group is aryl C such as benzyl, diphenylmethyl, triphenylmethyl, phenethyl, 2-phenylpropyl, 3-phenylpropyl and naphthylmethyl_1-6An alkyl group.

Aromatic C_1-6Alkoxy refers to aryl C such as benzyloxy, diphenylmethoxy, triphenylmethoxy, phenethyloxy, 2-phenylpropoxy, 3-phenylpropoxy and naphthylmethoxy_1-6An alkoxy group.

Aromatic C_1-6Alkoxy radical C_1-6The alkyl group is an aryl group C such as benzyloxymethyl and phenethyloxymethyl_1-6Alkoxy radical C_1-6An alkyl group.

Aromatic C_1-6The alkoxycarbonyl group is an arylC group such as benzyloxycarbonyl or phenethyloxycarbonyl_1-6An alkoxycarbonyl group.

The monocyclic nitrogen-containing heterocyclic group means a monocyclic nitrogen-containing heterocyclic group in which hetero atoms forming the above ring include only nitrogen atoms, such as aziridinyl, azetidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, piperidyl, tetrahydropyridinyl, dihydropyridinyl, pyridyl, homopiperidinyl, azocinyl, imidazolidinyl, imidazolinyl, imidazolyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, piperazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, homopiperazinyl, triazolyl, and tetrazolyl.

The monocyclic oxygen-containing heterocyclic group means a monocyclic oxygen-containing heterocyclic group containing only an oxygen atom as a hetero atom forming the above ring, such as an oxetanyl group, a tetrahydrofuryl group, a furyl group, a tetrahydropyranyl group, a pyranyl group, a1, 3-dioxanyl group, and a1, 4-dioxanyl group.

Monocyclic sulfur-containing heterocyclic group refers to thienyl.

The monocyclic nitrogen-and oxygen-containing heterocyclic group means a monocyclic nitrogen-and oxygen-containing heterocyclic group containing only a nitrogen atom and an oxygen atom as heteroatoms forming the above ring, such as an oxazolyl group, an isoxazolyl group, an oxadiazolyl group, a morpholinyl group, and an oxazepanyl group.

The monocyclic nitrogen-and sulfur-containing heterocyclic group means a monocyclic nitrogen-and sulfur-containing heterocyclic group containing only a nitrogen atom and a sulfur atom as the hetero atom forming the above ring, such as thiazolyl, isothiazolyl, thiadiazolyl, thiomorpholinyl, 1-oxide thiomorpholinyl, and 1, 1-dioxide thiomorpholinyl.

Monocyclic heterocyclic group means a monocyclic nitrogen-containing heterocyclic group, a monocyclic oxygen-containing heterocyclic group, a monocyclic sulfur-containing heterocyclic group, a monocyclic nitrogen-and sulfur-containing oxacyclic group or a monocyclic nitrogen-and sulfur-containing heterocyclic group.

The bicyclic nitrogen-containing heterocyclic group means a bicyclic nitrogen-containing heterocyclic group in which the hetero atom forming the above ring contains only a nitrogen atom, such as indolinyl, indolyl, isoindolinyl, isoindolyl, benzimidazolyl, indazolyl, benzotriazolyl, pyrazolopyridinyl, quinolyl, tetrahydroquinolyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, quinolizinyl, cinnolinyl, phthalazinyl, quinazolinyl, dihydroquinoxalinyl, quinoxalinyl, naphthyridinyl, purinyl, pteridinyl, and quinuclidinyl.

The bicyclic oxygen-containing heterocyclic group means a bicyclic oxygen-containing heterocyclic group containing only an oxygen atom as a heteroatom forming the above ring, such as a2, 3-dihydrobenzofuranyl group, benzofuranyl group, isobenzofuranyl group, chromanyl group, chromenyl group, isochroman group, 1, 3-benzodioxolyl group, 1, 3-benzodioxanyl group, and 1, 4-benzodioxanyl group.

The bicyclic sulfur-containing heterocyclic group means a bicyclic sulfur-containing heterocyclic group containing only a sulfur atom as a hetero atom forming the above ring, such as a2, 3-dihydrobenzothiophenyl group and a benzothiophenyl group.

The bicyclic nitrogen-and oxygen-containing heterocyclic group means a bicyclic nitrogen-and oxygen-containing heterocyclic group in which hetero atoms forming the above ring include only a nitrogen atom and an oxygen atom, such as a benzoxazolyl group, a benzisoxazolyl group, a benzoxadiazolyl group, a benzomorpholinyl group, a dihydropyranopyridyl group, a dioxazolyl group, a furopyridyl group, a dihydrodioxazinyl group, and a dihydropyridazinyl (dihydropyridoxazinyl) group.

The bicyclic nitrogen/sulfur-containing heterocyclic group means a bicyclic nitrogen/sulfur-containing heterocyclic group containing a nitrogen atom and a sulfur atom as heteroatoms forming the above ring, such as benzothiazolyl, benzisothiazolyl, and benzothiadiazolyl.

The bicyclic heterocyclic group means a bicyclic nitrogen-containing heterocyclic group, a bicyclic oxygen-containing heterocyclic group, a bicyclic sulfur-containing heterocyclic group, a bicyclic nitrogen-containing oxacyclic group or a bicyclic nitrogen-containing thiacyclic group.

The spiro heterocyclic group means a spiro heterocyclic group containing a nitrogen atom, an oxygen atom or a sulfur atom as a hetero atom forming the above ring, such as a 2-oxa-6-azaspiro [3.3] heptyl group, a1, 4-dioxaspiro [4.5] decyl group, a 1-oxa-8-azaspiro [4.5] decyl group, a 1-thia-8-azaspiro [4.5] decyl group and a 1-thia-8-azaspiro [4.5] decyl group.

The crosslinked heterocyclic group means a crosslinked heterocyclic group containing a nitrogen atom, an oxygen atom or a sulfur atom as a hetero atom forming the above-mentioned ring, such as a 3-oxa-8-azabicyclo [3.2.1] octyl group, an 8-oxa-3-azabicyclo [3.2.1] octyl group, a quinuclidine group or the like.

The heterocyclic group means a monocyclic heterocyclic group, a bicyclic heterocyclic group, a spiro heterocyclic group or a crosslinked heterocyclic group.

The heterocyclic oxy group is a substituent having an oxygen atom bonded to a heterocyclic group such as a pyrrolyloxy group, a piperidinyloxy group, a tetrahydrofuryloxy group, a tetrahydropyranyloxy group, or a tetrahydrothiopyranyloxy group.

The 5-to 10-membered nitrogen-containing phosphorus heterocycle refers to 5-to 10-membered nitrogen-containing phosphorus heterocycles which may be fused and include only nitrogen and phosphorus atoms as heteroatoms forming the above ring, such as 1, 3, 2-diazaphospholidine, 1, 3, 2-diazaphosphocyclohexane, 1, 3, 2-diazaphosphocycloheptane, and 1, 3, 2-diazaphosphocyclooctane.

The 5-to 10-membered oxygen-containing hetero ring is a 5-to 10-membered oxygen-containing hetero ring which may be condensed and contains only an oxygen atom and a phosphorus atom as hetero atoms forming the ring, such as 1, 3, 2-dioxaphospholane, 1, 3, 2-dioxaphosphorinane, 1, 3, 2-dioxaphosphocycloheptane, 1, 3, 2-dioxacyclooctane, benzo [ d ] [1, 3, 2] dioxaphosphole (dioxaphosphole), and 4H-benzo [ d ] [1, 3, 2] dioxaphosphinite.

The 5-to 10-membered nitrogen-oxygen-phosphorus-containing heterocycle is a 5-to 10-membered nitrogen-oxygen-phosphorus-containing heterocycle which may be condensed and which contains only a nitrogen atom, an oxygen atom and a phosphorus atom as heteroatoms forming the above ring, such as 1, 3, 2-oxazaphosphoridine, 2, 3-dihydrobenzo [ d ] [1, 3, 2] oxazaphosphorene, 1, 3, 2-oxazaphosphorane and 3, 4-dihydro-4H-benzo [ e ] [1, 3, 2] oxazaphosphine.

The 6-to 10-membered oxygen-containing hetero ring refers to 6-to 10-membered oxygen-containing hetero rings which may be condensed and contain only an oxygen atom and a phosphorus atom as hetero atoms forming the above rings, such as 1, 3, 2-dioxaphosphorinane, 1, 3, 2-dioxacycloheptane, and 1, 3, 2-dioxacyclooctane.

C_2-6The alkanoyl group is a straight or branched C group such as acetyl, propionyl, valeryl, isovaleryl or pivaloyl_2-6An alkanoyl group.

C_3-8Cycloalkylcarbonyl is C such as cyclopropylcarbonyl, cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl and cycloheptylcarbonyl_3-8A cycloalkyl carbonyl group.

Aroyl refers to benzoyl or naphthoyl, and the like.

The heterocyclic carbonyl group is a heterocyclic carbonyl group such as a pyrrolylcarbonyl group, a pyridylcarbonyl group, a furanylcarbonyl group or a thienylcarbonyl group.

Acyl refers to formyl, succinyl, glutaryl, maleoyl, phthaloyl, C_2-6Alkanoyl radical, C_3-8Cycloalkyl carbonyl, aroyl or heterocyclic carbonyl.

C_2-6The alkanoyloxy group is a straight or branched C group such as acetoxy, propionyloxy, valeryloxy, isovaleryloxy and pivaloyloxy_2-6An alkanoyloxy group.

C_3-8The cycloalkyl carbonyloxy group is C such as cyclopropylcarbonyloxy, cyclobutylcarbonyloxy, cyclopentylcarbonyloxy, cyclohexylcarbonyloxy and cycloheptylcarbonyloxy_3-8A cycloalkyl carbonyloxy group.

The aryloyloxy group means benzoyloxy group, naphthoyloxy group or the like.

The heterocyclic carbonyloxy group is a heterocyclic carbonyloxy group such as pyrrolylcarbonyloxy, pyridylcarbonyloxy, furylcarbonyloxy or thienylcarbonyloxy.

Acyloxy means C_2-6Alkanoyloxy group, C_3-8Cycloalkyl carbonyloxy, aroyloxy or heterocyclic carbonyloxy.

C_2-6The alkanoylthio group means a straight chain or branched chain such as acetylthio, propionylthio, valerylthio, isovalerylthio and pivaloylthioC of (A)_2-6An alkanoylthio group.

C_3-8The cycloalkylcarbonylthio group means C such as cyclopropylcarbonylthio, cyclobutylcarbonylthio, cyclopentylcarbonylthio, cyclohexylcarbonylthio and cycloheptylcarbonylthio_3-8A cycloalkylcarbonylthio group.

Aroylthio means benzoylthio or naphthoylthio and the like.

The heterocyclic carbonylthio group means a heterocyclic carbonylthio group such as pyrrolylcarbonylthio group, pyridylcarbonylthio group, furanylcarbonylthio group or thiophenylcarbonylthio group.

Acylthio means C_2-6Alkanoylthio group, C_3-8Cycloalkylcarbonylthio, arylylthio or heterocyclylcarbonylthio.

Silyl means trimethylsilyl, triethylsilyl, tributylsilyl or tert-butylmethylsilyl.

The leaving group is a halogen atom, C_1-6An alkylsulfonyloxy group, an aryloxy group or an arylsulfonyloxy group. C_1-6The alkylsulfonyloxy, aryloxy and arylsulfonyloxy radicals may be selected from halogen atoms, nitro radicals, C_1-6Alkyl and C_1-61 or more substituents in the alkoxy group.

Examples of the hydroxyl-protecting group include all groups that can be used as a protecting group for a hydroxyl group, and examples thereof include Protective group Organic Synthesis 4 th edition such as T.W. Greene, pp.16 to 299, 2007, John Wiley&Groups described in Sons, inc. Specific examples thereof include C_1-6Alkyl radical, C_2-6Alkenyl, aryl C_1-6Alkyl radical, C_1-6Alkoxy radical C_1-6Alkyl, aryl substituted C_1-6Alkoxy radical C_1-6Alkyl, acyl, C_1-6Alkoxycarbonyl, arylo C_1-6Alkoxycarbonyl group, C_1-6Alkylsulfonyl, arylsulfonyl, silyl, tetrahydrofuranyl, or tetrahydropyranyl.

The aliphatic hydrocarbon is pentane, hexane, heptane, cyclohexane, methylcyclohexane or ethylcyclohexane.

The halogenated hydrocarbon is dichloromethane, chloroform or dichloroethane.

Ethers are diethyl ether, diisopropyl ether, dioxane, tetrahydrofuran, anisole, ethylene glycol dimethyl ether, diethylene glycol dimethyl ether or diethylene glycol diethyl ether.

Ketones are acetone, 2-butanone, 4-methyl-2-pentanone or methyl isobutyl ketone.

The esters are methyl acetate, ethyl acetate, propyl acetate or butyl acetate.

Amides are N, N-dimethylformamide, N-dimethylacetamide or N-methylpyrrolidone.

Nitrile means acetonitrile or propionitrile.

Sulfoxides are dimethyl sulfoxide or sulfolane.

The aromatic hydrocarbon means benzene, toluene or xylene.

The inorganic base is selected from sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, sodium tert-butoxide, potassium tert-butoxide, sodium bicarbonate, sodium carbonate, potassium carbonate, tripotassium phosphate, potassium acetate, cesium fluoride, cesium carbonate or tert-butylmagnesium chloride.

The organic base is triethylamine, N-diisopropylethylamine, 1, 8-diazabicyclo (5.4.0) undec-7-ene (DBU), pyridine, 4-dimethylaminopyridine, N-methylmorpholine or imidazole.

Each substituent group has the following meaning.

<Substituent group A>A halogen atom; a hydroxyl group which may be protected; a cyano group; a nitro group; a carbamoyl group; an oxo group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkyl group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6Alkyl dithioA group; c which may be substituted by 1 or more substituents selected from substituent group B_2-6An alkenyl group; c which may be substituted by 1 or more substituents selected from substituent group B_2-6An alkynyl group; c which may be substituted by 1 or more substituents selected from substituent group B_3-8A cycloalkyl group; c which may be substituted by 1 or more substituents selected from substituent group B_3-8Cycloalkyl disulfide group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkoxy group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkoxycarbonyloxy group; c which may be substituted by 1 or more substituents selected from substituent group B_3-8A cycloalkoxycarbonyl group; aryl which may be substituted with 1 or more substituents selected from substituent group B; an aryl disulfide group which may be substituted with 1 or more substituents selected from substituent group B; a heterocyclic group which may be substituted with 1 or more substituents selected from substituent group B; a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group B; an acyloxy group which may be substituted by 1 or more substituents selected from the substituent group B; an acylthio group which may be substituted with 1 or more substituents selected from substituent group B; aminocarbonyloxy which may be substituted with 1 or more substituents selected from substituent group B; or aminocarbonylthio which may be substituted by 1 or more substituents selected from substituent group B

<Substituent group B>A halogen atom; a hydroxyl group; a cyano group; a nitro group; a carboxyl group; a carbamoyl group; a hydroxymethyl group; c_1-6An alkyl group; c_2-6An alkenyl group; c_2-6An alkynyl group; c_3-8A cycloalkyl group; c_1-6An alkoxy group; c_1-6An alkoxycarbonyl group; c_3-8A cycloalkoxycarbonyl group; aromatic C_1-6An alkoxycarbonyl group; an aryl group; an aryloxy group; heterocyclic oxy which may be substituted with 1 or more substituents selected from hydroxy and hydroxymethyl; aromatic C_1-6An alkoxy group; or acyloxy group

Examples of the salt of the thionucleoside derivative represented by the general formula [1] include salts of a basic group such as an amino group and an acidic group such as a hydroxyl group and a carboxyl group, which are generally known.

Examples of the salt of a basic group include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, and sulfuric acid; salts with organic carboxylic acids such as formic acid, acetic acid, citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid, malic acid, tartaric acid, aspartic acid, trichloroacetic acid, and trifluoroacetic acid; and with methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid,Sulfonic acid salts such as sulfonic acid and naphthalenesulfonic acid.

Examples of the salt of the acidic group include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; an ammonium salt; and salts with nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl- β -phenethylamine, 1-diphenylmethylamine, and N, N' -dibenzylethylenediamine.

Among the above salts, preferable salts include pharmaceutically acceptable salts.

Prevention refers to inhibition of onset, reduction in risk of onset, delay of onset, or the like.

The treatment refers to the suppression of the improvement or deterioration of the disease or condition to be treated.

The treatment refers to prevention or treatment of various diseases.

The treatment agent is a substance supplied for the purpose of prevention, treatment, or the like of various diseases.

A benign tumor is a tumor in which the tumor cells and their arrangement take a morphology close to that of the normal cells from which they are derived, and which is not invasive or metastatic.

Malignant tumors are tumors in which the morphology or arrangement of tumor cells is different from that of normal cells from which they are derived, and which show infiltrative or metastatic properties.

The tumor refers to benign tumor or malignant tumor.

The thionucleoside derivative or the salt thereof of the present invention can be used for the treatment of tumors.

The thionucleoside derivative or a salt thereof of the present invention is preferably used for, for example, treatment of lung cancer, esophageal cancer, stomach cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, kidney cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma, or malignant lymphoma.

In other ways, the thionucleoside derivative or salt thereof of the present invention is preferably used for obtaining gemcitabine resistant tumor treatment.

The thionucleoside derivative of the present invention is represented by the general formula [1 ].

[ chemical formula 2]

(in the formula, R¹、R²And R³Has the same meaning as described above. )

¹R

R¹Is hydroxy which may be protected, C which may be substituted_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted.

As R¹Preferably, the hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6Alkoxy, aryloxy which may be substituted with 1 or more substituents selected from substituent group a, heterocyclic oxy which may be substituted with 1 or more substituents selected from substituent group a, or amino which may be substituted with 1 or more substituents selected from substituent group a, more preferablyC which may be substituted by 1 or more substituents selected from substituent group A_1-6Alkoxy, aryloxy which may be substituted with 1 or more substituents selected from substituent group a, or amino which may be substituted with 1 or more substituents selected from substituent group a, and further preferably C which may be substituted with 1 or more substituents selected from substituent group a_1-6An alkoxy group or an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a.

As R¹C which may be substituted with 1 or more substituents selected from substituent group A_1-6The substituent of the alkoxy group is preferably C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkyl disulfide group, C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkoxycarbonyloxy, acyloxy which may be substituted by 1 or more substituents selected from substituent group B, or acylthio which may be substituted by 1 or more substituents selected from substituent group B, more preferably C which may be substituted by 1 or more substituents selected from substituent group B_1-6Alkyl disulfide group, C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkoxycarbonyloxy, C which may be substituted with 1 or more substituents selected from substituent group B_2-6Alkanoyloxy or C which may be substituted with 1 or more substituents selected from substituent group B_2-6An alkanoylthio group.

Wherein C is optionally substituted by 1 or more substituents selected from substituent group B_1-6Substituents of alkyldithio groups, preferably hydroxy, C_1-6Alkoxy, aryl C_1-6Alkoxy or acyloxy, more preferably hydroxy, C_1-6Alkoxy, aryl C_1-6Alkoxy or C_2-6An alkanoyloxy group.

As R¹The substituent of the aryloxy group which may be substituted with 1 or more substituents selected from the substituent group A, preferably a halogen atom or C which may be substituted with 1 or more substituents selected from the substituent group B_1-6An alkoxy group.

As R¹The substituent of the amino group which may be substituted with 1 or more substituents selected from the substituent group A, preferably C which may be substituted with 1 or more substituents selected from the substituent group B_1-6An alkyl group.

Wherein C is optionally substituted by 1 or more substituents selected from substituent group B_1-6Substituents of alkyl, preferably C_1-6Alkoxycarbonyl, aryl or arylC_1-6An alkoxycarbonyl group.

²R

R²Is C which may be substituted_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted.

As R²Preferably C which may be substituted by 1 or more substituents selected from substituent group A_1-6Alkoxy, aryloxy which may be substituted with 1 or more substituents selected from substituent group a, or amino which may be substituted with 1 or more substituents selected from substituent group a, more preferably C which may be substituted with 1 or more substituents selected from substituent group a_1-6An alkoxy group or an amino group which may be substituted with 1 or more substituents selected from substituent group a.

As R²C which may be substituted with 1 or more substituents selected from substituent group A_1-6The substituent of the alkoxy group is preferably C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkyl disulfide group, C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkoxycarbonyloxy, heterocyclic oxy which may be substituted with 1 or more substituents selected from substituent group B, acyloxy which may be substituted with 1 or more substituents selected from substituent group B, acylthio which may be substituted with 1 or more substituents selected from substituent group B, aminocarbonyloxy which may be substituted with 1 or more substituents selected from substituent group B, orAminocarbonylthio which may be substituted with 1 or more substituents selected from substituent group B, more preferably C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkyl disulfide group, C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkoxycarbonyloxy, acyloxy which may be substituted by 1 or more substituents selected from substituent group B, or acylthio which may be substituted by 1 or more substituents selected from substituent group B, and further preferably C which may be substituted by 1 or more substituents selected from substituent group B_1-6Alkyl disulfide group, C which may be substituted with 1 or more substituents selected from substituent group B_1-6Alkoxycarbonyloxy, C which may be substituted with 1 or more substituents selected from substituent group B_2-6Alkanoyloxy or C which may be substituted with 1 or more substituents selected from substituent group B_2-6An alkanoylthio group.

Wherein C is optionally substituted by 1 or more substituents selected from substituent group B_1-6Substituents of alkyldithio groups, preferably hydroxy, aryl C_1-6Alkoxy or acyloxy, more preferably hydroxy, aryl C_1-6Alkoxy or C_1-6An alkanoyloxy group.

As the substituent of the heterocyclic oxy group which may be substituted with 1 or more substituents selected from the substituent group B, a hydroxyl group or a hydroxymethyl group is preferable.

As R²The substituent of the amino group which may be substituted with 1 or more substituents selected from the substituent group A, preferably C which may be substituted with 1 or more substituents selected from the substituent group B_1-6An alkyl group or a heterocyclic group which may be substituted with 1 or more substituents selected from substituent group B.

Wherein C is optionally substituted by 1 or more substituents selected from substituent group B_1-6Alkyl substituents, preferably carboxy, C_1-6Alkoxycarbonyl group, C_3-8Cycloalkoxycarbonyl, aryl or arylC_1-6An alkoxycarbonyl group.

R¹And R²Or may form a 5-to 10-membered nitrogen-containing/phosphorus-containing heterocycle which may be substituted, a 5-to 10-membered oxygen-containing/phosphorus-containing heterocycle which may be substituted, or a 5-to 10-membered nitrogen-containing/oxygen-phosphorus-containing heterocycle which may be substituted, together with the phosphorus atom to which they are bonded.

As R¹And R²The ring formed together with the phosphorus atom to which they are bonded is preferably a 5-to 10-membered oxygen-containing phosphorus heterocycle which may be substituted, more preferably 1, 3, 2-dioxaphosphorinane or 4H-benzo [ d ]][1，3，2]A dioxaphosphinite.

As R¹And R²A substituent of a 5-to 10-membered optionally substituted nitrogen-containing/phosphorus-containing heterocycle, an optionally substituted 5-to 10-membered optionally substituted oxygen-containing/phosphorus-containing heterocycle or an optionally substituted 5-to 10-membered optionally substituted nitrogen-containing/oxygen-phosphorus-containing heterocycle, which is formed together with the phosphorus atom bonded thereto, is preferably C which may be substituted with 1 or more substituents selected from substituent group A_1-6An alkoxy group or an aryl group which may be substituted with 1 or more substituents selected from substituent group a.

Among them, as the substituent of the aryl group which may be substituted with 1 or more substituents selected from the substituent group a, a halogen atom is preferable.

³R

R³Is a hydrogen atom.

R²And R³Or with R²Bonded phosphorus atom, oxygen atom, methylene group, 2 carbon atoms constituting tetrahydrothiophene ring and R³The bonded oxygen atoms form together a 6-to 10-membered oxygen-containing phosphorus heterocycle which may be substituted.

As R²And R³And R²Bonded phosphorus atom, oxygen atom, methylene group, 2 carbon atoms constituting tetrahydrothiophene ring and R³The bonded oxygen atoms together form a ring, preferably 1, 3, 2-dioxaphosphorinane.

As a general formula [1]The compound shown is preferably R¹Is hydroxy which may be protected, C which may be substituted_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted; r²Is C which may be substituted_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted; r³Compounds which are hydrogen atoms, more preferably R¹A hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a; r²Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a; r³A compound which is a hydrogen atom.

When isomers (for example, tautomers, optical isomers, geometric isomers, and the like) exist in the thionucleoside derivative represented by the general formula [1] or a salt thereof, the present invention includes the isomers thereof, and also includes solvates, hydrates, and crystals of various shapes.

Next, a method for producing the compound of the present invention will be described.

The compound of the present invention can be produced by combining known methods, and can be produced, for example, by the following production methods.

[ production method A ]

[ chemical formula 3]

(in the formula, R^aRepresents a silyl group which may be substituted; r^bRepresents optionally substituted C_1-20Alkoxycarbonyl or optionally substituted C_3-8A cycloalkoxycarbonyl group; x¹Represents a leaving group. )

First step of

As the compound of the general formula [ S1], for example, t-butyldiphenylchlorosilane is known.

The compound of the general formula [ a2] can be produced by reacting a compound of the general formula [ a1] with a compound of the general formula [ S1] in the presence of a base.

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include aliphatic hydrocarbons, halogenated hydrocarbons, ethers, ketones, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons, and these solvents may be used in combination.

Preferred solvents include amides, and more preferably N, N-dimethylformamide.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the formula [ A1 ].

The amount of the compound of the general formula [ S1] used may be 1 to 5 times by mol, preferably 1 to 2 times by mol, based on the compound of the formula [ A1 ].

The base used in this reaction may be an inorganic base or an organic base, preferably an organic base, and more preferably imidazole.

The amount of the base used may be 1 to 10 times by mol, preferably 1 to 5 times by mol, based on the compound of the formula [ a1 ].

The reaction is carried out at 0 to 100 ℃ and preferably 10 to 50 ℃ for 30 minutes to 48 hours.

Second step of

Examples of known compounds of the general formula [ S2] include di-tert-butyl dicarbonate.

The compound of the general formula [ A3] can be produced by reacting a compound of the general formula [ a2] with a compound of the general formula [ S2] in the presence of a base.

Preferred solvents include halogenated hydrocarbons, and dichloromethane is more preferred.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A2 ].

The amount of the compound of the general formula [ S2] used may be 3 to 10 times by mol, preferably 3 to 6 times by mol, based on the compound of the general formula [ A2 ].

Examples of the base used in the reaction include inorganic bases and organic bases, preferably organic bases, and more preferably pyridine, triethylamine and diisopropylethylamine. These bases may also be used in combination.

The amount of the base used may be 3 to 20 times by mol, preferably 3 to 10 times by mol, based on the compound of the general formula [ A2 ]. Further, a base may be used as the solvent.

Preferably, a reaction promoter is added to the reaction.

Examples of the reaction accelerator include 4-dimethylaminopyridine.

The amount of the reaction accelerator to be used may be 0.01 to 1 time by mol based on the compound of the general formula [ A2 ].

Third Process

The compound of the general formula [ a4] can be produced by deprotecting a compound of the general formula [ A3 ].

This reaction can be carried out by the method described in, for example, Protective group in Organic Synthesis 4 th edition, pages 16 to 366, 2007, John Wiley & Sons, INC.

[ production method B ]

[ chemical formula 4]

(in the formula, R^cRepresents an optionally substituted acyl group, an optionally substituted C_1-20Alkoxycarbonyl or optionally substituted C_3-8A cycloalkoxycarbonyl group; x²Represents a leaving group; r^aHas the same meaning as described above. )

First step of

As the compound of the general formula [ S3], for example, benzoyl chloride is known.

The compound of the general formula [ a5] can be produced by reacting a compound of the general formula [ a2] with a compound of the general formula [ S3] in the presence of a base.

Preferred solvents include halogenated hydrocarbons, ethers and amides, and dichloromethane, tetrahydrofuran and N, N-dimethylformamide are more preferred.

The amount of the compound of the general formula [ S3] used may be 2 to 20 times by mol, preferably 2 to 5 times by mol, based on the compound of the general formula [ A2 ].

The amount of the base used may be 2 to 20 times by mol, preferably 2 to 10 times by mol, based on the compound of the general formula [ A2 ]. Further, a base may be used as the solvent.

In this reaction, a reaction accelerator may be added as needed.

Examples of the reaction accelerator include 4-dimethylaminopyridine.

The reaction is carried out at 0 to 100 ℃ and preferably 0 to 40 ℃ for 30 minutes to 48 hours.

Second step of

The compound of the general formula [ a6] can be produced by deprotecting a compound of the general formula [ a5 ].

This reaction may be carried out according to the third step of production method a.

[ production method C ]

[ chemical formula 5]

(in the formula, R^b、R^c、X²Has the same meaning as described above. )

First step of

The compound of the general formula [ a7] can be produced by reacting a compound of the general formula [ a1] with a compound of the general formula [ S2] in the presence of a base.

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include ethers and water, and these solvents may be used in combination.

Preferred examples of the solvent include tetrahydrofuran, 1, 4-dioxane and water, and more preferred examples include a mixed solvent of tetrahydrofuran and water and a mixed solvent of 1, 4-dioxane and water.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A1 ].

The amount of the compound of the general formula [ S2] used may be 2 to 20 times by mol, preferably 2 to 10 times by mol, based on the compound of the general formula [ A1 ].

The base used in this reaction may be an inorganic base or an organic base, preferably an inorganic base, and more preferably sodium hydroxide or potassium hydroxide. These bases may also be used in combination.

The amount of the base used may be 2 to 30 times by mol, preferably 2 to 15 times by mol, based on the compound of the general formula [ A1 ]. Further, a base may be used as the solvent.

In this reaction, a reaction accelerator may be added as needed.

Examples of the reaction accelerator include 4-dimethylaminopyridine.

The amount of the reaction accelerator to be used may be 0.01 to 1 time by mol based on the compound of the general formula [ A1 ].

Second step of

The compound of the general formula [ A8] can be produced by reacting a compound of the general formula [ a7] with a compound of the general formula [ S3] in the presence of a base.

This reaction may be carried out according to the first step of production method B.

Third Process

The compound of the general formula [ a9] can be produced by deprotecting a compound of the general formula [ A8 ].

[ production method 1]

[ chemical formula 6]

(in the formula, R^1aAnd R^2aThe same or different, represent C which may be substituted_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted; or R^1aAnd R^2aOr may form a 5-to 10-membered nitrogen-containing/phosphorus-containing heterocycle which may be substituted, a 5-to 10-membered oxygen-containing/phosphorus-containing heterocycle which may be substituted, or a 5-to 10-membered nitrogen-containing/oxygen-phosphorus-containing heterocycle which may be substituted, together with the phosphorus atom to which they are bonded; x³Represents a leaving group; r^bHas the same meaning as described above. )

First step of

As the compound of the general formula [ S4], for example, (2S) -benzyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) propionate, (2S) -benzyl 2- (((RS) -chloro (4-chlorophenoxy) phosphoryl) amino) propionate, S- (2- (((RS) -chloro (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate, and (2RS, 4R) -2- (4-nitrophenoxy) -4-phenyl-1, 3, 2-dioxaphosphorinane 2-oxide are known.

The compound of the general formula [ a10] can be produced by reacting a compound of the general formula [ a4] with a compound of the general formula [ S4] in the presence of a base.

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include ethers and aromatic hydrocarbons, and these solvents may be used in combination.

Preferred solvents include ethers, and tetrahydrofuran is more preferred.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A4 ].

The amount of the compound of the general formula [ S4] used may be 1 to 20 times by mol, preferably 1 to 10 times by mol, based on the compound of the general formula [ A4 ].

Examples of the base used in this reaction include tert-butyl magnesium chloride.

The amount of the base used may be 1 to 5 times by mol, preferably 1 to 2 times by mol, based on the compound of the general formula [ A4 ].

The reaction is carried out at-78 to 100 ℃ and preferably-78 to 40 ℃ for 30 minutes to 48 hours.

Second step of

The compound of the general formula [1a ] can be produced by deprotecting a compound of the general formula [ A10 ].

[ production method 2]

[ chemical formula 7]

(in the formula, R^1bAnd R^2bThe same or different, represent C which may be substituted_1-20Alkoxy or C which may be substituted_3-8A cycloalkoxy group; r^dAnd R^eThe same or different, represent C which may be substituted_1-20An alkyl group; r^bHas the same meaning as described above. )

First step of

As the compound of the general formula [ S5], for example, S' - (((((diisopropylamino) phosphinodiyl) bis (oxy)) bis (ethane-2, 1-diyl)) bis (2, 2-dimethylthiopropionate) and S- (2- (((2-cyanoethoxy) (diisopropylamino) phosphino) oxy) ethyl) 2, 2-dimethylthiopropionate are known.

The compound of the general formula [ a11] can be produced by reacting a compound of the general formula [ a4] with a compound of the general formula [ S5] and then reacting the reaction product with an oxidizing agent.

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include halogenated hydrocarbons, ethers, amides, and nitriles, and these solvents may be used in combination.

Preferred solvents include dichloromethane, tetrahydrofuran, N-dimethylformamide and acetonitrile, and dichloromethane is more preferred.

The amount of the compound of the general formula [ S5] used may be 1 to 10 times by mol, preferably 1 to 5 times by mol, based on the compound of the general formula [ A4 ].

Preferably, a reaction promoter is added to the reaction.

Examples of the reaction accelerator include 1H-tetrazole.

The amount of the reaction accelerator to be used is preferably 1 to 10 times by mol, more preferably 1 to 5 times by mol, based on the compound of the general formula [ A4 ].

The reaction with the compound of the general formula [ S5] may be carried out at-78 to 40 ℃ for 30 minutes to 24 hours.

Examples of the oxidizing agent used in this reaction include m-chloroperbenzoic acid.

The amount of the oxidizing agent to be used may be 1 to 5 times by mol, preferably 1 to 2 times by mol, based on the compound of the general formula [ A4 ].

The reaction with the oxidant is carried out at-78 to 40 ℃ for 30 minutes to 24 hours.

Second step of

The compound of the general formula [1b ] can be produced by deprotecting a compound of the general formula [ A11 ].

[ production method 3]

[ chemical formula 8]

(in the formula, R^1cRepresents optionally substituted C_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group or an amino group which may be substituted; x⁴Represents a leaving group; r^bHas the same meaning as described above. )

First step of

As the compound of the general formula [ S6], for example, phosphorus oxychloride is known.

As the compound of the general formula [ S7], for example, benzyl L-alanine and 2, 2' -disulfanediyldiethanol are known.

The compounds of the general formulae [ a12] and [ a13] can be produced by reacting a compound of the general formula [ a4] with a compound of the general formula [ S6] in the presence of a base, and then reacting the compound with a compound of the general formula [ S7 ].

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include halogenated hydrocarbons, ethers, and aromatic hydrocarbons, and these solvents may be used in combination.

The amount of the compound of the general formula [ S6] used may be 1 to 3 times by mol, preferably 1 to 2 times by mol, based on the compound of the general formula [ A4 ].

The base used in this reaction is preferably an organic base, and more preferably triethylamine and diisopropylethylamine. These bases may also be used in combination.

The amount of the base used is preferably 3 to 15 times by mol, more preferably 3 to 6 times by mol, based on the compound of the general formula [ A4 ].

The reaction with the compound of the general formula [ S6] may be carried out at-78 to 0 ℃ for 30 minutes to 24 hours.

The amount of the compound of the general formula [ S7] used may be 1 to 5 times by mol, preferably 2 to 3 times by mol, based on the compound of the general formula [ A4 ].

The reaction with the compound of the general formula [ S7] may be carried out at-78 to 0 ℃ for 30 minutes to 24 hours.

The reaction with the compound of the general formula [ S7] may be carried out at 0 to 100 ℃ and preferably 0 to 40 ℃ for 30 minutes to 24 hours.

Second step of

The compounds of the general formula [1c ] and the general formula [1d ] can be produced by deprotecting compounds of the general formula [ A12] and the general formula [ A13 ].

[ production method 4]

[ chemical formula 9]

(in the formula, R^4aRepresents optionally substituted C_1-20An alkyl group; x⁵Represents a leaving group; r^bAnd R^1cHas the same meaning as described above. )

First step of

As the compound of the general formula [ S8], chloromethyl isopropyl carbonate and chloromethyl pivalate are known, for example.

The compound of the general formula [ a14] can be produced by reacting a compound of the general formula [ a12] with a compound of the general formula [ S8] in the presence of a base.

The solvent used in the reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include amides, nitriles, and sulfoxides, and these solvents may be used in combination.

Preferred solvents include amides, and more preferably N, N-dimethylformamide.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A12 ].

The amount of the compound of the general formula [ S8] used may be 1 to 10 times by mol, preferably 1 to 5 times by mol, based on the compound of the general formula [ A12 ].

Examples of the base used in the reaction include an inorganic base and an organic base, preferably an organic base, and more preferably triethylamine and N, N-diisopropylethylamine. These bases may also be used in combination.

The amount of the base used may be 1 to 20 times by mol, preferably 1 to 10 times by mol, based on the compound of the general formula [ A12 ]. Further, a base may be used as the solvent.

The reaction is carried out at 0 to 100 ℃ and preferably 50 to 80 ℃ for 30 minutes to 48 hours.

Second step of

The compound of the general formula [1e ] can be produced by deprotecting a compound of the general formula [ A14 ].

[ production method 5]

[ chemical formula 10]

(in the formula, R^4a、R^b、X⁴And X⁵Has the same meaning as described above. )

First step of

The compound of the general formula [ a15] can be produced by reacting a compound of the general formula [ a4] with a compound of the general formula [ S6] in the presence of a base.

The compound of the formula [ A15] is preferably used directly in the next step without isolation.

Second step of

The compound of the general formula [ a16] can be produced by reacting a compound of the general formula [ a15] with water in the presence of a base.

The amount of the base used may be 2 to 10 times by mol, preferably 2 to 4 times by mol, based on the compound of the general formula [ A15 ].

The reaction is carried out at 0 to 50 ℃ for 30 minutes to 24 hours.

Third Process

The compounds of the general formulae [ a17] and [ a18] can be produced by reacting a compound of the general formula [ a16] with a compound of the general formula [ S8] in the presence of a base.

Preferred solvents include amides, and more preferably N, N-dimethylformamide.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A16 ].

The amount of the compound of the general formula [ S8] used may be 2 to 20 times by mol, preferably 2 to 10 times by mol, based on the compound of the general formula [ A16 ].

The amount of the base used may be 2 to 20 times by mol, preferably 2 to 10 times by mol, based on the compound of the general formula [ A16 ]. Further, a base may be used as the solvent.

The fourth step

The compounds of the general formulae [1f ] and [1g ] can be produced by deprotecting compounds of the general formulae [ A17] and [ A18 ].

[ production method 6]

[ chemical formula 11]

(in the formula, R^fRepresents optionally substituted C_1-20An alkoxy group; r^c、R^eAnd R^1cHas the same meaning as described above. )

First step of

As the compound of the general formula [ S9], for example, 2-cyanoethyl N, N, N ', N' -tetraisopropyl phosphoramidite is known.

The compound of the general formula [ a19] can be produced by reacting a compound of the general formula [ a9] with a compound of the general formula [ S9] and then reacting the reaction product with an oxidizing agent.

The amount of the solvent used is not particularly limited, and may be 1 to 50 times (v/w) the amount of the compound of the general formula [ A9 ].

The amount of the compound of the general formula [ S9] used may be 1 to 10 times by mol, preferably 1 to 5 times by mol, based on the compound of the general formula [ A9 ].

Preferably, a reaction promoter is added to the reaction.

Examples of the reaction accelerator include 1H-tetrazole.

The amount of the reaction accelerator to be used is preferably 2 to 10 times by mol, more preferably 2 to 5 times by mol, based on the compound of the general formula [ A9 ].

The reaction with the compound of the general formula [ S9] may be carried out at-78 to 40 ℃ for 30 minutes to 24 hours.

The amount of the oxidizing agent to be used may be 1 to 5 times by mol, preferably 1 to 2 times by mol, based on the compound of the general formula [ A9 ].

Second step of

The compound of the formula [1h ] can be produced by deprotecting a compound of the general formula [ A19 ].

Third Process

As the compound of the general formula [ S7], for example, L-alanine benzyl ester and isopropyl alcohol are known.

The compound of the general formula [1i ] can be produced by reacting the compound of the general formula [1h ] with an activator, and then reacting the compound with the compound of the general formula [ S7 ].

The amount of the solvent used is not particularly limited, as long as it is 1 to 50 times (v/w) the amount of the compound of the formula [1h ].

Examples of the activator include oxalyl chloride.

The amount of the activating agent to be used is only 1 to 5 times by mol, preferably 1 to 2 times by mol, based on the compound of the formula [1h ].

The reaction with the activator may be carried out at-78 to 40 ℃ and preferably-78 to 0 ℃ for 30 minutes to 24 hours.

The amount of the compound of the general formula [ S7] used is only required to be 1 to 5 times by mol, preferably 2 to 3 times by mol, relative to the compound of the formula [1h ].

Preferably, a reaction promoter is added to the reaction.

Examples of the reaction accelerator include N, N-dimethylformamide.

The amount of the reaction accelerator to be used is preferably 0.01 to 1 time by mol based on the compound of the formula [1h ].

The compound obtained by the above production method can be derived into another compound by, for example, imparting a self-known reaction such as condensation, addition, oxidation, reduction, rearrangement, substitution, halogenation, dehydration or hydrolysis, or by appropriately combining these reactions.

When an amino group, a hydroxyl group or a carboxyl group is present in the compound obtained by the above production method and an intermediate thereof, these protecting groups may be appropriately substituted to carry out the reaction. When the compound has 2 or more protecting groups, deprotection can be selectively performed by a reaction known per se.

Among the compounds used in the above production methods, compounds which can be in the form of salts may be used as salts. Examples of the salts include the same salts as those of the thionucleoside derivative represented by the general formula [1] which is the thionucleoside derivative of the present invention.

When an isomer (for example, tautomer, optical isomer, geometric isomer, or the like) exists in the compound used in the above production method, these isomers may also be used. When a solvate, a hydrate, and a crystal having various shapes are present, these solvate, hydrate, and crystal having various shapes may be used.

The pharmaceutical composition containing the thionucleoside derivative represented by the general formula [1] or a salt thereof of the present invention may be appropriately mixed with additives usually used for pharmaceutical preparations.

Examples of the additives include excipients, disintegrants, binders, lubricants, flavoring agents, colorants, flavoring agents, surfactants, coating agents, and plasticizers.

Examples of the excipient include sugar alcohols such as erythritol, mannitol, xylitol, and sorbitol; sugars such as white sugar, powdered sugar, lactose, and glucose; cyclodextrins such as α -cyclodextrin, β -cyclodextrin, γ -cyclodextrin, hydroxypropyl β -cyclodextrin, and sodium sulfobutylether β -cyclodextrin; celluloses such as crystalline cellulose and microcrystalline cellulose; and starches such as corn starch, potato starch and gelatinized starch.

Examples of the disintegrant include carboxymethylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, sodium carboxymethyl starch, crospovidone, low-substituted hydroxypropyl cellulose, and partially gelatinized starch.

Examples of the binder include hydroxypropyl cellulose, sodium carboxymethyl cellulose, and methyl cellulose.

Examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, talc, hydrated silicon dioxide, light anhydrous silicic acid, and sucrose fatty acid ester.

Examples of the taste-imparting agent include aspartame, saccharin, stevioside, thaumatin and acesulfame potassium.

Examples of the coloring agent include titanium dioxide, ferric oxide, yellow ferric oxide, black ferric oxide, edible red No. 102, edible yellow No. 4, and edible yellow No. 5.

Examples of the flavoring agent include essential oils such as orange oil, lemon oil, peppermint oil, and pine oil; essence such as orange essence and peppermint essence; cherry essence, vanilla essence, fruit essence and other essences; powdery flavors such as apple-, banana-, peach-, strawberry-and orange-colored fine particles; vanillin; and ethyl vanillin.

Examples of the surfactant include sodium lauryl sulfate, dioctyl sodium sulfosuccinate, polysorbate, and polyoxyethylene hydrogenated castor oil.

Examples of the coating agent include hydroxypropylmethylcellulose, aminoalkyl methacrylate copolymer E, aminoalkyl methacrylate copolymer RS, ethylcellulose, cellulose acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid copolymer L, methacrylic acid copolymer LD, and methacrylic acid copolymer S.

Examples of the plasticizer include triethyl citrate, polyethylene glycol, triacetin, and propylene glycol.

These additives may be used alone or in combination of two or more.

The amount to be blended is not particularly limited, and may be appropriately blended in accordance with the respective purposes so as to sufficiently exhibit the effects thereof.

The pharmaceutical composition to which the mixture is appropriately added may be administered orally or parenterally in the form of tablets, capsules, powders, syrup preparations, granules, pills, suspensions, emulsions, liquids, powder preparations, suppositories, eye drops, nose drops, ear drops, patches, ointments, injections or the like according to a conventional method, and preferably orally in the form of tablets, capsules, powders, syrup preparations, granules, pills, suspensions, emulsions, liquids, powder preparations or the like.

The method, amount and frequency of administration of the thionucleoside derivative represented by the general formula [1] or a salt thereof of the present invention can be appropriately selected depending on the age, weight and symptoms of a patient. In general, the administration of 0.01 to 1000mg/kg orally or parenterally is performed 1 to several times per 1 day for an adult, and preferably, the administration of 0.01 to 1000mg/kg orally is performed 1 to several times per 1 day.

Examples

The present invention will be described with reference to reference examples and examples, but the present invention is not limited to them.

When not specifically described, the purification by column chromatography is performed by using an automatic purification apparatus ISOLERA (manufactured by Biotage corporation) or medium pressure liquid chromatography YFLC-Wprep2XY.N (manufactured by Shanghan corporation).

Unless otherwise specified, SNAPKP-Sil Cartridge (manufactured by Biotage corporation), Hi-Flash Columns W001, W002, W003, W004 and W005 (manufactured by Shanghai Co., Ltd.) were used as the carrier in silica gel column chromatography, and SNAPKP-NH Cartridge (manufactured by Biotage corporation) was used as the carrier in alkaline silica gel column chromatography.

Thin layer chromatography for fractionation using PLC glass plate silica gel F₆₀(Merck Co., Ltd.).

Reversed phase fractionation HPLC was performed using Waters 2998Photodiode Array (PDA) Detector (manufactured by Waters), Waters 600Controller (manufactured by Waters), Waters 2767 Sample Manager (manufactured by Waters), and YMC-Actus ProC18, 30X 50mm column (manufactured by YMC).

The mixing ratio of the eluent is the volume ratio. For example, "gradient elution of hexane and ethyl acetate of 100: 0 to 50: 50" is an elution in which 100% hexane/0% ethyl acetate is finally changed to 50% hexane/50% ethyl acetate.

MS spectra were measured using an ACQUITY SQD LC/MS System (manufactured by Waters, Ionization method: ESI (ElectroSpray Ionization)), model M-8000 (manufactured by Hitachi, Ionization method: ESI method), or LCMS-2010EV (manufactured by Shimadzu, Ionization method: Ionization method in which ESI and APCI (atmospheric pressure Ionization) were performed simultaneously).

NMR spectra were measured using BrukeraV300 (300 MHz, manufactured by Bruker Co.) using tetramethylsilane as an internal standard, and all values are expressed in ppm.

The Retention Time (RT) was measured in minutes (min) using SQD (Waters corporation).

Column: BEHC 181.7 μm, 2.1X30mm (made by Waters corporation)

Solvent: solution A: 0.1% formic acid-water

And B, liquid B: 0.1% formic acid-acetonitrile

Gradient circulation: 0.00min (liquid a/B95/5), 2.00min (liquid a/B5/95), 3.00min (liquid a/B5/95), 3.01min (liquid a/B100/0), 3.80min (liquid a/B100/0)

Flow rate: 0.5mL/min

Column temperature: at room temperature

Detection wavelength: 254nm

In each example, each abbreviation has the following meaning.

Ac: acetyl group

Boc: tert-butoxycarbonyl group

TBDPS: tert-butyldiphenylsilyl

Ts: p-toluenesulfonyl (p-toluenesulfonyl)

RT (min): retention time (minutes)

DMSO-d₆: deuterated dimethyl sulfoxide

*: bonding position

Reference example 1

[ chemical formula 12]

First step of

To a mixture of methanesulfonic acid salt of 4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (10.0g) and N, N-dimethylformamide (100mL) were added imidazole (5.70g) and tert-butyldiphenylchlorosilane (11.5g), and the mixture was stirred at room temperature for 1.5 hours. Water was added to the reaction solution, followed by extraction with ethyl acetate, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give 4-amino-1- ((2R, 3S, 4S, 5R) -5- (((tert-butyldiphenylsilyl) oxy) methyl) -3-fluoro-4-hydroxytetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (13.8g) as a colorless oil.

MS(ESI m/z)：500(M+H)

RT(min)：1.50

Second step of

To a mixture of 4-amino-1- ((2R, 3S, 4S, 5R) -5- (((tert-butyldiphenylsilyl) oxy) methyl) -3-fluoro-4-hydroxytetrahydrothien-2-yl) pyrimidin-2 (1H) -one (13.8g) obtained in the first step and dichloromethane (100mL) were added N, N-dimethyl-4-aminopyridine (100mg) and triethylamine (12.5mL), and a mixture of di-tert-butyl dicarbonate (20mL) and dichloromethane (20mL) was slowly added dropwise at room temperature. After stirring at room temperature for 6 hours, triethylamine (5.0mL) and di-tert-butyl dicarbonate (5.0mL) were added and the mixture was stirred for 20 minutes. To the reaction mixture were added triethylamine (3.0mL) and di-tert-butyl dicarbonate (3.0mL), and the mixture was stirred for 30 minutes, followed by distillation under reduced pressure to remove the solvent. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 100: 0 to 70: 30) to give tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -5- (((tert-butyldiphenylsilyl) oxy) methyl) -3-fluorotetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (21.8g) as a white solid.

MS(ESI m/z)：800(M+H)

RT(min)：2.52

Third Process

To a mixture of tert-butyltert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -5- (((tert-butyldiphenylsilyl) oxy) methyl) -3-fluorotetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (14.8g) and tetrahydrofuran (30mL) was added a mixture of acetic acid (3.0mL) and 1mol/L tetrabutylammonium fluoride/tetrahydrofuran solution (30mL), and the mixture was stirred at room temperature for 45 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 100: 0 to 50: 50) to give tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (7.50g) as a white solid.

MS(ESI m/z)：562(M+H)

RT(min)：1.71

Reference example 2

[ chemical formula 13]

First step of

To a mixture of 4-amino-1- ((2R, 3S, 4S, 5R) -5- (((tert-butyldiphenylsilyl) oxy) methyl) -3-fluoro-4-hydroxytetrahydrothien-2-yl) pyrimidin-2 (1H) -one (11.5g) and pyridine (115mL) was added benzoyl chloride (7.94mL) under ice-cooling, and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and water was added to the obtained residue to conduct extraction with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure and purified by silica gel column chromatography (hexane: ethyl acetate: 90: 10 to 0: 100) to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((tert-butyldiphenylsilyl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (8.48g) as a white solid.

MS(ESI m/z)：708(M+H)

RT(min)：2.28

Second step of

To a mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((tert-butyldiphenylsilyl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoate (8.48g) and tetrahydrofuran (48mL) was added 1mol/L tetrabutylammonium fluoride/tetrahydrofuran solution (24mL), and the mixture was stirred at room temperature for 30 minutes. Methylene chloride was added to the reaction solution, and the organic layer was washed with water and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate: 90: 10 to 0: 100) to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- (hydroxymethyl) tetrahydrothiophene-3-benzoic acid ester (5.56g) as a white solid.

¹H-NMR(DMSO-d₆)：11.36(s，1H)，8.71(d，1H，J＝7.6Hz)，8.05-7.99(m，4H)，7.76-7.45(m，7H)，6.60(dd，1H，J＝12.4，5.4Hz)，5.86-5.79(m，1H)，5.71-5.48(m，2H)，3.82-3.68(m，3H).

MS(ESI m/z)：470(M+H)

RT(min)：1.24

Reference example 3

[ chemical formula 14]

First step of

To a mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- (hydroxymethyl) tetrahydrothiophene-3-benzoate (9.50g), 1H-tetrazole (4.30g), and dichloromethane (190mL) was added di-tert-butyl N, N-diisopropylphosphoramidite (12.8mL), and the mixture was stirred at room temperature for 30 minutes. M-chloroperbenzoic acid (7.00g) was added at-40 ℃ and stirred under ice-cooling for 30 minutes. A mixture of sodium sulfite (7.70g) and water (100mL) was added under ice-cooling, and stirred at room temperature for 30 minutes. The aqueous layer was removed, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 50: 50 to 20: 80) to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((di-tert-butoxyphosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (4.93g) as a white solid.

¹H-NMR(DMSO-d₆)：11.38(s，1H)，8.56(d，1H，J＝7.9Hz)，8.05-8.00(m，4H)，7.76-7.71(m，1H)，7.67-7.45(m，6H)，6.62(dd，1H，J＝14.4，5.1Hz)，5.92-5.83(m，1H)，5.76-5.53(m，1H)，4.37-4.17(m，2H)，3.97-3.89(m，1H)，1.42(s，9H)，1.41(s，9H).

MS(ESI m/z)：662(M+H)

RT(min)：1.71

Second step of

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((di-tert-butoxyphosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (3.50g) and 4.0mol/L hydrogen chloride in 1, 4-dioxane (27mL) was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was washed with ethyl acetate to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- ((phosphonooxy) methyl) tetrahydrothiophene-3-benzoic acid ester (2.63g) as a white solid.

¹H-NMR(DMSO-d₆)：8.61(d，1H，J＝7.6Hz)，8.05-8.00(m，4H)，7.77-7.70(m，1H)，7.68-7.49(m，5H)，7.45(d，1H，J＝7.6Hz)，6.62(dd，1H，J＝13.5，5.3Hz)，5.87-5.80(m，1H)，5.74-5.54(m，1H)，4.31-4.12(m，2H)，3.91(dd，1H，J＝11.1，6.1Hz).

MS(ESI m/z)：550(M+H)

RT(min)：0.92

Reference example 4

[ chemical formula 15]

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- ((phosphonooxy) methyl) tetrahydrothiophene-3-benzoic acid ester (1.70g) and 7.0mol/L ammonia/methanol solution (100mL) was stirred at room temperature for 17 hours. Insoluble matter was filtered off, and the solvent was distilled off under reduced pressure. The resulting residue was washed with acetone to give ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyl dihydrogen phosphate (733mg) as a white solid.

¹H-NMR(DMSO-d₆)：8.00(d，1H，J＝7.6Hz)，7.31(s，1H)，7.17(s，1H)，6.42(dd，1H，J＝11.2，5.3Hz)，5.80(d，1H，J＝7.6Hz)，5.04-4.80(m，1H)，4.34-4.22(m，1H)，3.97-3.79(m，2H)，3.34-3.26(m，1H).

MS(ESI m/z)：342(M+H)

RT(min)：0.19

Reference example 5

[ chemical formula 16]

First step of

To a mixture of the mesylate salt of 4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (2.40g) and 1.0mol/L aqueous potassium hydroxide solution (160mL) was slowly added a mixture of di-tert-butyl dicarbonate (17.4g) and 1, 4-dioxane (160mL) over 20 minutes. After completion of the dropwise addition, the mixture was stirred at room temperature for 3.5 hours, and then 1.0mol/L aqueous potassium hydroxide solution (80mL) and di-tert-butyl dicarbonate (8.72g) were added to the reaction mixture, followed by stirring at room temperature for 30 minutes. After standing at room temperature for 15 hours, the mixture was extracted with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 90: 10) to give tert-butyl ((2R, 3S, 4S, 5R) -3- ((tert-butoxycarbonyl) oxy) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluorotetrahydrothiophen-2-yl) methyl carbonate (1.35g) as a colorless oil.

MS(ESI m/z)：462(M+H)

RT(min)：1.29

Second step of

To a mixture of tert-butyl ((2R, 3S, 4S, 5R) -3- ((tert-butoxycarbonyl) oxy) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluorotetrahydrothiophen-2-yl) methyl carbonate (50.0mg), N-diisopropylethylamine (36.7. mu.L) and dichloromethane (1.0mL) was added benzoyl chloride (18.8. mu.L), and the mixture was stirred at room temperature for 18 hours. To the reaction mixture was added a saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, ethyl acetate was added to the obtained residue, and the precipitated solid was collected by filtration to give tert-butyl (((2R, 3S, 4S, 5R) -3- ((tert-butoxycarbonyl) oxy) -4-fluoro-5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) tetrahydrothiophen-2-yl) methyl) carbonate (38.0mg) as a white solid.

MS(ESI m/z)：566(M+H)

RT(min)：1.78

Third Process

Trifluoroacetic acid (1.0mL) was added to tert-butyl (((2R, 3S, 4S, 5R) -3- ((tert-butoxycarbonyl) oxy) -4-fluoro-5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) tetrahydrothiophen-2-yl) methyl) carbonate (31.0mg), and the mixture was stirred at room temperature for 3 hours. The solvent was distilled off under reduced pressure, and ethyl acetate (1.0mL) and a saturated aqueous solution of sodium hydrogencarbonate (1.0mL) were added to the resulting residue and stirred. The aqueous layer was removed, the organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol: 95: 5 to 88: 12) to obtain N- (1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (9.2mg) as a white solid.

¹H-NMR(DMSO-d₆)：11.31(s，1H)，8.63(d，1H，J＝7.6Hz)，8.01(d，2H，J＝7.6Hz)，7.67-7.59(m，1H)，7.56-7.47(m，2H)，7.40(d，1H，J＝7.3Hz)，6.47(dd，1H，J＝10.9，5.6Hz)，5.96(d，1H，J＝5.3Hz)，5.38-5.30(m，1H)，5.20-4.95(m，1H)，4.34-4.21(m，1H)，3.83-3.62(m，2H)，3.30-3.21(m，1H).

MS(ESI m/z)：366(M+H)

RT(min)：0.88

Reference example 6

[ chemical formula 17]

To a mixture of diphenyl phosphite (272. mu.L) and pyridine (1.8mL) was added a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (400mg) and pyridine (1.8mL), and the mixture was stirred at room temperature for 10 minutes. A mixture of water (712. mu.L) and triethylamine (712. mu.L) was added thereto, and the mixture was stirred at room temperature for 5 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (chloroform: methanol: 100: 0 to 60: 40) to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylhydrogenphosphonate (283mg) as a colorless oil.

MS(ESI m/z)：626(M+H)

RT(min)：1.26

Reference example 7

[ chemical formula 18]

To a mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- (hydroxymethyl) tetrahydrothiophene-3-benzoate (3.90g), 2-cyanoethyl N, N, N ', N' -tetraisopropyl phosphorodiamidite (5.00g), N, N-diisopropylethylamine (2.2mL), acetonitrile (28mL) and tetrahydrofuran (28mL) was added 1H-tetrazole (870mg), and the mixture was stirred at room temperature for 20 minutes. Water was added to the reaction solution, extracted with ethyl acetate, and the organic layer was dried over anhydrous magnesium sulfate and then the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 50 to 0: 100) to obtain a white solid. The obtained white solid was dissolved in methanol and ethyl acetate, and hexane was added thereto to precipitate the white solid, thereby obtaining (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((((2-cyanoethoxy) (diisopropylamino) phosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (1.50g) as a white solid.

¹H-NMR(DMSO-d₆)：11.37(s，1H)，8.70-8.54(m，1H)，8.07-7.39(m，10H)，6.66-6.55(m，1H)，5.93-5.80(m，1H)，5.76-5.66(m，1H)，5.60-5.48(m，1H)，4.12-3.70(m，5H)，3.65-3.48(m，2H)，2.85-2.74(m，2H)，1.18-1.09(m，12H).

Reference example 8-1

[ chemical formula 19]

First step of

Pivaloyl chloride (2.64mL) was slowly added dropwise to a mixture of 2-mercaptoethanol (1.5mL), triethylamine (4.0mL) and methylene chloride (55mL) at-78 ℃ over 1 hour, and the mixture was stirred at room temperature for 1 hour. Water (30mL) was added to the reaction mixture, and after stirring for 30 minutes, the mixture was extracted with ethyl acetate, and the organic layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give S- (2-hydroxyethyl) 2, 2-dimethylthiopropionate (3.69g) as a colorless oil.

¹H-NMR(CDCl₃)：3.76(t，2H，J＝6.1Hz)，3.06(t，2H，J＝6.1Hz)，1.25(s，9H).

Second step of

To a mixture of S- (2-hydroxyethyl) 2, 2-dimethylthiopropionate (1.62g), triethylamine (3.0mL) and tetrahydrofuran (25mL) was added dropwise a mixture of 1, 1-dichloro-N, N-diisopropylphosphinamine (1.01g) and tetrahydrofuran (35mL) at-78 ℃ and stirred at room temperature for 2 hours. After insoluble matter was filtered off, the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 96: 4 to 75: 25) to obtain S, S' - (((((diisopropylamino) phosphinediyl) bis (oxy)) bis (ethane-2, 1-diyl)) bis (2, 2-dimethylthiopropionate) (751mg) as a colorless oil.

¹H-NMR(CDCl₃)：4.24(t，4H，J＝7.3Hz)，3.79-3.59(m，2H)，2.89-2.76(m，4H)，1.27-1.15(m，30H).

Reference example 8-2

[ chemical formula 20]

The following compounds were obtained in the same manner as in reference example 8-1.

S, S' - ((((diisopropylamino) phosphinediyl) bis (oxy)) bis (ethane-2, 1-diyl)) bis (2-methylthiopropionate)

¹H-NMR(CDCl₃)：4.25(t，4H，J＝6.9Hz)，2.89-2.77(m，4H)，2.63-2.47(m，4H)，1.24-1.14(m，24H).

Reference examples 8 to 3

[ chemical formula 21]

S, S' - (((((diisopropylamino) phosphinediyl) bis (oxy)) bis (ethane-2, 1-diyl)) dithiopropionate

¹H-NMR(CDCl₃)：4.26(t，4H，J＝6.9Hz)，3.79-3.60(m，2H)，2.90-2.73(m，4H)，1.19(d，12H，J＝6.6Hz)，1.14(t，6H，J＝7.6Hz).

Reference example 9-1

[ chemical formula 22]

To a mixture of S- (2-hydroxyethyl) 2, 2-dimethylthiopropionate (500mg), pyridine (0.5mL) and methylene chloride (5.0mL) was added p-toluenesulfonyl chloride (881mg), and the mixture was stirred at room temperature for 19 hours. To the reaction mixture was added a saturated aqueous sodium bicarbonate solution (5.0mL), and the mixture was stirred for 1.5 hours and then extracted with ethyl acetate. The organic layer was washed with 1.0mol/L hydrochloric acid, water and a saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give S- (2- (p-toluenesulfonyloxy) ethyl) 2, 2-dimethylthiopropionate (762mg) as a colorless oil.

¹H-NMR(CDCl₃)：7.80(d，2H，J＝8.3Hz)，7.35(d，2H，J＝8.3Hz)，4.08(t，2H，J＝6.6Hz)，3.08(t，2H，J＝6.6Hz)，2.45(s，3H)，1.18(s，9H).

Reference example 9-2

[ chemical formula 23]

The following compounds were obtained in the same manner as in reference example 9-1.

S- (4- (p-toluenesulfonyloxy) butyl) 2, 2-dimethylthiopropionate

MS(ESI m/z)：345(M+H)

RT(min)：1.90

Reference example 10-1

[ chemical formula 24]

To a mixture of 2, 2' -disulfanediyldiethanol (500mg) and tetrahydrofuran (5.0mL) was added 60% sodium hydride (130mg) under a nitrogen atmosphere, and the mixture was stirred at room temperature for 10 minutes. Methyl iodide (202. mu.L) was added to the reaction mixture, and after stirring for 3 hours, water was added thereto and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 72: 28 to 51: 49) to give 2- ((2-methoxyethyl) disulfide) ethanol (187mg) as a colorless oil.

¹H-NMR(CDCl₃)：3.95-3.85(m，2H)，3.67(t，2H，J＝6.3Hz)，3.39(s，3H)，2.91(t，2H，J＝6.3Hz)，2.88(t，2H，J＝5.6Hz)，2.23-2.15(m，1H).

Reference example 10-2

[ chemical formula 25]

The following compounds were obtained in the same manner as in reference example 10-1.

2- ((2- (benzyloxy) ethyl) disulfide) ethanol

¹H-NMR(CDCl₃)：7.39-7.27(m，5H)，4.56(s，2H)，3.92-3.81(m，2H)，3.75(t，2H，J＝6.3Hz)，2.94(t，2H，J＝6.3Hz)，2.83(t，2H，J＝5.9Hz)，2.12-1.95(m，1H).

Reference example 11-1

[ chemical formula 26]

First step of

Ethyl isocyanate (1.1mL) was added to a mixture of 2-mercaptoethanol (980. mu.L), triethylamine (3.9mL) and toluene (14mL) at-78 ℃ and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure to give S- (2-hydroxyethyl) ethylthiocarbamate (2.10 g).

Second step of

To a mixture of S- (2-hydroxyethyl) ethylthiocarbamate (1.05g), pyridine (1.1mL) and dichloromethane (14mL) was added p-toluenesulfonyl chloride (2.00g) under ice-cooling, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was added with a saturated aqueous sodium bicarbonate solution, stirred for 30 minutes, and then extracted with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give 2- ((ethylcarbamoyl) thio) ethyl 4-methylbenzenesulfonate (470 mg).

MS(ESI m/z)：304(M+H)

RT(min)：1.35

Reference example 11-2

[ chemical formula 27]

The following compounds were obtained in the same manner as in reference example 11-1.

2- ((isopropylcarbamoyl) thio) ethyl 4-methylbenzenesulfonate

MS(ESI m/z)：318(M+H)

RT(min)：1.46

Reference examples 11 to 3

[ chemical formula 28]

2- ((cyclohexylcarbamoyl) thio) ethyl 4-methylbenzenesulfonate

MS(ESI m/z)：358(M+H)

RT(min)：1.60

Reference example 12-1

[ chemical formula 29]

To a mixture of 2-cyanoethyl N, N-diisopropylphosphoramidite (223. mu.L) and diethyl ether (4.0mL) under a nitrogen atmosphere, a mixture of N, N-diisopropylethylamine (172. mu.L), 1-dodecanol (224. mu.L) and diethyl ether (6.0mL) was added dropwise under ice cooling. After stirring at room temperature for 2.5 hours, the insoluble matter was filtered off, and the solvent was distilled off under reduced pressure to give 2-cyanoethyldodecylN, N-diisopropylphosphoramidite (298mg) as a colorless oil.

Reference example 12-2

[ chemical formula 30]

The following compounds were obtained in the same manner as in reference example 12-1.

2-cyanoethyl-2-ethylhexyl diisopropylphosphoramidite

Reference examples 12 to 3

[ chemical formula 31]

2-cyanoethyloctadecyldiisopropylphosphoramidite

Reference example 13-1

[ chemical formula 32]

To a mixture of 4-nitrophenyldichlorophosphate (5.12g) and tetrahydrofuran (51mL) was added pyridine (4.8mL) under ice-cooling. After stirring for 30 minutes under ice-cooling, 1, 3-propanediol (1.52g) was added under ice-cooling, and the mixture was stirred at room temperature for 1 hour. Saturated aqueous ammonium chloride was added thereto, and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 20: 80) to obtain 2- (4-nitrophenoxy) -1, 3, 2-dioxaphosphorinane 2-oxide (2.74g) as a white solid.

MS(ESI m/z)：260(M+H)

RT(min)：0.98

Reference example 13-2

[ chemical formula 33]

The following compounds were obtained in the same manner as in reference example 13-1.

(2RS, 4R) -2- (4-Nitrophenoxy) -4-phenyl-1, 3, 2-dioxaphosphorinane 2-oxide

MS(ESI m/z)：336(M+H)

RT(min)：1.39

Reference examples 13 to 3

[ chemical formula 34]

(2RS, 4RS) -4- (3-chlorophenyl) -2- (4-nitrophenoxy) -1, 3, 2-dioxaphosphorinane 2-oxide

MS(ESI m/z)：370(M+H)

RT(min)：1.51

Reference examples 13 to 4

[ chemical formula 35]

(RS) -2- (4-Nitrophenoxy) -4H-benzo [ d ] [1, 3, 2] dioxaphosphino 2-oxide

MS(ESI m/z)：308(M+H)

RT(min)：1.36

Reference examples 13 to 5

[ chemical formula 36]

(RS) -8-methoxy-2- (4-nitrophenoxy) -4H-benzo [ d ] [1, 3, 2] dioxaphosphino 2-oxide

MS(ESI m/z)：338(M+H)

RT(min)：1.34

Reference example 14-1

[ chemical formula 37]

To a mixture of ethylene glycol (11.4mL), t-butyldiphenylchlorosilane (10.6mL) and methylene chloride (80mL) was added imidazole (8.30g), and the mixture was stirred at room temperature for 25 minutes. Water was added to the reaction solution, followed by extraction with dichloromethane, and the organic layer was dried over anhydrous magnesium sulfate, followed by distillation under reduced pressure to remove the solvent. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate 95: 5 to 60: 40) to obtain 2- ((tert-butyldiphenylsilyl) oxy) ethanol (3.87g) as a colorless oil.

¹H-NMR(CDCl₃)：7.74-7.33(m，10H)，3.80-3.73(m，2H)，3.72-3.64(m，2H)，1.07(s，9H).

Reference example 14-2

[ chemical formula 38]

The following compounds were obtained in the same manner as in reference example 14-1.

2- ((2- ((tert-butyldiphenylsilyl) oxy) ethyl) dithio) ethanol

¹H-NMR(CDCl₃)：7.70-7.36(m，10H)，3.90(t，2H，J＝6.6Hz)，3.87-3.79(m，2H)，2.85(t，2H，J＝6.6Hz)，2.75(t，2H，J＝5.9Hz)，1.90(t，1H，J＝5.9Hz)，1.06(s，9H).

Reference example 15

[ chemical formula 39]

To a mixture of 2, 3, 4, 6-tetra-O-acetyl-. beta. -D-glucopyranose (100mg) and dichloromethane (1.5mL) was added trichloroacetonitrile (115. mu.L) under ice-cooling, and the mixture was stirred for 5 minutes. Diazabicycloundecene (10. mu.L) was added to the reaction solution, stirred at room temperature for 40 minutes, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 65: 35) to obtain (2R, 3R, 4S, 5R, 6S) -2- (acetoxymethyl) -6- (2, 2, 2-trichloro-1-iminoethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (95.0mg) as a colorless oil.

¹H-NMR(CDCl₃)：8.70(s，1H)，6.56(d，1H，J＝4.0Hz)，5.62-5.51(m，1H)，5.22-5.07(m，2H)，4.33-4.06(m，3H)，2.12-1.96(m，12H).

Reference example 16-1

[ chemical formula 40]

First step of

To a mixture of (2R, 3R, 4S, 5R, 6S) -2- (acetoxymethyl) -6- (2, 2, 2-trichloro-1-iminoethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (525mg), 2- ((tert-butyldiphenylsilyl) oxy) ethanol (318mg), molecular sieves 4A and dichloromethane (5.3mL) was added boron trifluoride diethyl ether complex (54. mu.L) under ice-cooling, and the mixture was stirred at room temperature for 10 minutes. Water was added to the reaction solution, followed by extraction with methylene chloride. The organic layer was dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 50: 50) to obtain (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((tert-butyldiphenylsilyl) oxy) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (170mg) as a white solid.

¹H-NMR(CDCl₃)：7.71-7.33(m，10H)，5.26-4.96(m，3H)，4.63(d，1H，J＝7.9Hz)，4.30-4.21(m，1H)，4.14-4.07(m，1H)，3.94-3.61(m，5H)，2.13-1.93(m，12H)，1.03(s，9H).

Second step of

To a mixture of (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((tert-butyldiphenylsilyl) oxy) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (170mg), acetic acid (31. mu.L) and tetrahydrofuran (2.7mL) was added 1mol/L tetrabutylammonium fluoride/tetrahydrofuran solution (540. mu.L), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 50 to 0: 100) to give (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2-hydroxyethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (79.0mg) as a colorless oil.

¹H-NMR(CDCl₃)：5.30-5.16(m，1H)，5.11-4.97(m，2H)，4.56(d，1H，J＝7.9Hz)，4.20(d，2H，J＝4.0Hz)，3.88-3.66(m，5H)，2.13-1.97(m，12H).

Reference example 16-2

[ chemical formula 41]

First step of

To a mixture of (2R, 3R, 4S, 5R, 6S) -2- (acetoxymethyl) -6- (2, 2, 2-trichloro-1-iminoethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (525mg), 2- ((2- ((tert-butyldiphenylsilyl) oxy) ethyl) disulfide group) ethanol (417mg), molecular sieves 4A and dichloromethane (5.3mL) was added boron trifluoride diethyl ether complex (54. mu.L) under ice-cooling, and the mixture was stirred at room temperature for 10 minutes. Water was added to the reaction solution, followed by extraction with dichloromethane, and the organic layer was dried over anhydrous magnesium sulfate, followed by distillation under reduced pressure to remove the solvent. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 50: 50) to give (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2- ((tert-butyldiphenylsilyl) oxy) ethyl) disulfide) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (75.0mg) as a colorless oil.

¹H-NMR(CDCl₃)：7.70-7.34(m，10H)，5.24-4.94(m，3H)，4.51(d，1H，J＝8.9Hz)，4.31-3.99(m，3H)，3.92-3.64(m，4H)，2.86-2.71(m，4H)，2.15-1.98(m，12H)，1.06(s，9H).

Second step of

To a mixture of (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2- ((tert-butyldiphenylsilyl) oxy) ethyl) dithio) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (75.0mg), acetic acid (12. mu.L) and tetrahydrofuran (1.0mL) was added 1mol/L tetrabutylammonium fluoride/tetrahydrofuran solution (540. mu.L), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 50 to 0: 100) to give (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2-hydroxyethyl) disulfide) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (33.0mg) as a colorless oil.

¹H-NMR(CDCl₃)：5.26-5.17(m，1H)，5.14-5.05(m，1H)，5.04-4.96(m，1H)，4.57-4.54(m，1H)，4.32-2.82(m，11H)，2.06-2.02(m，12H).

Examples 1 to 1

[ chemical formula 42]

First step of

(1) To a mixture of phenyl dichlorophosphate (600. mu.L), L-alanine benzyl ester hydrochloride (864mg) and methylene chloride (20mL) was added triethylamine (1.1mL) at-78 ℃ and the mixture was stirred at room temperature for 2 hours and 11 minutes. The solvent was distilled off under reduced pressure, diethyl ether was added to the obtained residue, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -benzyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) propionate as a crude product.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (80.0mg) and tetrahydrofuran (2.0mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (710. mu.L) at-78 ℃ and stirred at room temperature for 10 minutes. A mixture of (2S) -benzyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) propionate (150mg) obtained in (1) and tetrahydrofuran (900. mu.L) was added to the reaction mixture, and the mixture was stirred at room temperature for 40 minutes. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate and drying over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to give (2S) -benzyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate.

MS(ESI m/z)：879(M+H)

RT(min)：2.06

Second step of

To a mixture of (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate and dichloromethane (1.4mL) was added trifluoroacetic acid (1.4mL) under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol: 100: 0 to 85: 15) to obtain a colorless oil. The resulting colorless oil was purified by reverse phase fractional HPLC (0.1% aqueous formic acid-0.1% acetonitrile formic acid) to give a formate salt of (2S) -benzyl 2- (((RS) - (((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate as a colorless oil (3.6 mg).

¹H-NMR(CD₃OD)：8.14(s，1H)，8.16-7.99(m，1H)，7.42-7.13(m，10H)，6.69-6.58(m，1H)，5.92-5.85(m，1H)，5.15(s，2H)，5.09-4.86(m，1H)，4.45-4.17(m，3H)，4.09-3.94(m，1H)，3.61-3.48(m，1H)，1.41-1.30(m，3H).

MS(ESI m/z)：579(M+H)

RT(min)：1.13

Examples 1 to 2

The compounds in tables 1 and 2 were obtained in the same manner as in example 1-1.

[ Table 1]

[ Table 2]

Example 2-1

[ chemical formula 43]

First step of

(1) To a mixture of phenyl dichlorophosphate (300. mu.L), L-phenylalanine benzyl ester hydrochloride (460mg) and methylene chloride (10mL) was added triethylamine (553. mu.L) at-78 ℃ and the mixture was stirred for 15 minutes and at room temperature for 1 hour. The solvent was distilled off under reduced pressure, diethyl ether was added to the obtained residue, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -ethyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) -3-phenylpropionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (80.0mg) and tetrahydrofuran (2.0mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L solution of tert-butylmagnesium chloride/tetrahydrofuran (710. mu.L) at-78 ℃ and stirred at room temperature for 10 minutes. A mixture of (2S) -ethyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) -3-phenylpropionate (150mg) obtained in (1) and tetrahydrofuran (900. mu.L) was added to the reaction mixture, and the mixture was stirred at room temperature for 4 hours and 45 minutes. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate and drying over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to give (2S) -ethyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) -3-phenylpropionate.

MS(ESI m/z)：893(M+H)

RT(min)：2.09

Second step of

To a mixture of (2S) -ethyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) -3-phenylpropionate and dichloromethane (1.4mL) was added trifluoroacetic acid (1.4mL) under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the resulting residue was purified by reverse phase fractional HPLC (0.1% aqueous formic acid-0.1% acetonitrile formic acid solution) to give a colorless oil. The colorless oil obtained was purified by silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 85: 15), to give (2S) -ethyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) -3-phenylpropionate (2.3mg) as a colorless oil.

¹H-NMR(CD₃OD)：8.05-7.97(m，1H)，7.36-7.05(m，10H)，6.70-6.58(m，1H)，5.91-5.83(m，1H)，5.07-4.86(m，1H)，4.39-3.40(m，7H)，3.15-3.01(m，1H)，2.93-2.80(m，1H)，1.22-1.13(m，3H).

MS(ESI m/z)：593(M+H)

RT(min)：1.14

Examples 2 to 2

[ chemical formula 44]

First step of

The following compounds were obtained in the same manner as in the first step of example 2-1.

(2S) -Ethyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) -4-methylpentanoate

MS(ESI m/z)：859(M+H)

RT(min)：2.12

Second step of

The following compounds were obtained in the same manner as in the second step of example 2-1.

(2S) -Ethyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) -4-methylpentanoate

¹H-NMR(CD₃OD)：8.18-8.01(m，1H)，7.41-7.14(m，5H)，6.74-6.59(m，1H)，5.93-5.86(m，1H)，5.10-4.88(m，1H)，4.50-3.52(m，7H)，1.81-1.68(m，1H)，1.59-1.45(m，2H)，1.27-1.19(m，3H)，0.96-0.77(m，6H).

MS(ESI m/z)：559(M+H)

RT(min)：1.15

Example 3-1

[ chemical formula 45]

First step of

(1) To a mixture of L-alanine benzyl ester hydrochloride (646mg), (4-chlorophenyl) phosphoryl dichloride (500. mu.L), and dichloromethane (1.5mL) was added triethylamine (827. mu.L) at-78 ℃ and stirred for 5 minutes, and at room temperature for 70 minutes. Hexane was added to the reaction mixture, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -benzyl 2- (((RS) -chloro (4-chlorophenoxy) phosphoryl) amino) propionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (448mg) and tetrahydrofuran (10.6mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (2.0mL) at-78 ℃ and stirred for 5 minutes. A mixture of (2S) -benzyl 2- (((RS) -chloro (4-chlorophenoxy) phosphoryl) amino) propionate obtained in (1) and tetrahydrofuran (5.3mL) was added to the reaction mixture at-78 ℃ and stirred at room temperature for 30 minutes. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate and drying over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 0: 100) to give (2S) -benzyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-chlorophenoxy) phosphoryl) amino) propionate (539mg) as a yellow solid.

MS(ESI m/z)：913(M+H)

RT(min)：2.13

Second step of

To a mixture of (2S) -benzyl 2- (((RS) - ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-chlorophenoxy) phosphoryl) amino) propionate (514mg) and dichloromethane (1.5mL), trifluoroacetic acid (1.5mL) was added, followed by stirring at room temperature for 40 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by basic silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 50: 50) to give (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophene-2-yl) methoxy) (4-chlorophenoxy) phosphoryl) amino) propionate (135mg) as a white solid.

¹H-NMR(CD₃OD)：8.08-8.00(m，1H)，7.38-7.13(m，9H)，6.71-6.59(m，1H)，5.91-5.83(m，1H)，5.16-4.84(m，3H)，4.45-4.16(m，3H)，4.09-3.93(m，1H)，3.62-3.47(m，1H)，1.40-1.34(m，3H).

MS(ESI m/z)：613(M+H)

RT(min)：1.23

Examples 3 to 2

[ chemical formula 46]

First step of

The following compounds were obtained in the same manner as in the first step of example 3-1.

(2S) -methyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate

MS(ESI m/z)：803(M+H)

RT(min)：1.89

Second step of

The following compounds were obtained in the same manner as in the second step of example 3-1.

(2S) -methyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-Azo-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) propanoate

¹H-NMR(CD₃OD)：8.15-8.05(m，1H)，7.42-7.16(m，5H)，6.73-6.61(m，1H)，5.95-5.87(m，1H)，5.12-4.83(m，1H)，4.50-4.23(m，3H)，4.05-3.91(m，1H)，3.72-3.53(m，4H)，1.40-1.29(m，3H).

MS(ESI m/z)：503(M+H)

RT(min)：0.87

Example 4-1

[ chemical formula 47]

First step of

(1) To a mixture of L-alanine cyclobutyl ester (148mg) and dichloromethane (1.5mL) were added phenyl dichlorophosphate (153. mu.L) and triethylamine (143. mu.L) at-78 ℃ and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, diethyl ether was added to the resulting residue, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -cyclobutyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) propionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (100mg) and tetrahydrofuran (2.0mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (500. mu.L) at-78 ℃ and stirred for 30 minutes. A mixture of (2S) -cyclobutyl 2- (((RS) -chloro (phenoxy) phosphoryl) amino) propionate obtained in (1) and tetrahydrofuran (2.0mL) was added to the reaction mixture at-78 ℃ and stirred at room temperature for 16 hours. Water was added to the reaction solution, which was extracted with ethyl acetate and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 100: 0 to 70: 30) to give (2S) -cyclobutyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate (33.7mg) as a colorless oil.

MS(ESI m/z)：843(M+H)

RT(min)：1.76

Second step of

To a mixture of (2S) -cyclobutyl 2- (((RS) - ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate (33.7mg) and dichloromethane (1.0mL) was added trifluoroacetic acid (1.0mL), and the mixture was stirred at room temperature for 2.5 hours. The solvent was distilled off under reduced pressure, and methylene chloride (2.0mL) and triethylamine (1.0mL) were added to the obtained residue, and the solvent was distilled off under reduced pressure. The resulting residue was purified by basic silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 80: 20) to give (2S) -cyclobutyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate as a white solid (11.3 mg).

¹H-NMR(CD₃OD)：8.13-8.00(m，1H)，7.42-7.32(m，2H)，7.29-7.15(m，3H)，6.72-6.60(m，1H)，5.93-5.84(m，1H)，5.11-4.89(m，2H)，4.49-4.21(m，3H)，3.99-3.86(m，1H)，3.66-3.51(m，1H)，2.39-2.24(m，2H)，2.15-1.97(m，2H)，1.86-1.71(m，1H)，1.70-1.58(m，1H)，1.39-1.27(m，3H).

MS(ESI m/z)：543(M+H)

RT(min)：1.09

Example 4 to 2

[ chemical formula 48]

First step of

The following compounds were obtained in the same manner as in the first step of example 4-1.

(2S) -cyclohexyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate

MS(ESI m/z)：871(M+H)

RT(min)：1.72

Second step of

The following compounds were obtained in the same manner as in the second step of example 4-1.

(2S) -cyclohexyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate

¹H-NMR(CD₃OD)：8.12-8.02(m，1H)，7.41-7.32(m，2H)，7.28-7.15(m，3H)，6.72-6.60(m，1H)，5.92-5.85(m，1H)，5.10-4.90(m，1H)，4.81-4.66(m，1H)，4.49-4.23(m，3H)，3.99-3.86(m，1H)，3.65-3.50(m，1H)，1.88-1.66(m，4H)，1.58-1.23(m，9H).

MS(ESI m/z)：571(M+H)

RT(min)：1.27

Example 5-1

[ chemical formula 49]

First step of

(1) To a mixture of phosphorus oxychloride (139. mu.L) and dichloromethane (15mL) was added 4-iodophenol (330mg) and triethylamine (207. mu.L) at-78 ℃ and the mixture was stirred for 5 minutes. The reaction mixture was stirred at room temperature for 10 minutes, and L-alanine benzyl ester hydrochloride (324mg) and triethylamine (415. mu.L) were added thereto at-78 ℃ and stirred for 5 minutes. After stirring at room temperature for 30 minutes, hexane was added to the reaction solution, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -benzyl 2- (((RS) -chloro (4-iodophenoxy) phosphoryl) amino) propionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (185mg) and tetrahydrofuran (4.5mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (833. mu.L) at-78 ℃ and stirred for 5 minutes. A mixture of (2S) -benzyl 2- (((RS) -chloro (4-iodophenoxy) phosphoryl) amino) propionate obtained in (1) and tetrahydrofuran (2.2mL) was added to the reaction mixture at-78 ℃ and stirred at room temperature for 30 minutes. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate and drying over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 80: 20 to 0: 100) to give (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-iodophenoxy) phosphoryl) amino) propionate (123mg) as a colorless oil.

MS(ESI m/z)：1005(M+H)

RT(min)：2.14

Second step of

Trifluoroacetic acid (1.0mL) was added to a mixture of (2S) -benzyl 2- (((RS) - ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-iodophenoxy) phosphoryl) amino) propionate (123mg) and dichloromethane (1.0mL), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by basic silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 50: 50) to give (2S) -benzyl 2- ((((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (4-iodophenoxy) phosphoryl) amino) propionate (68.0mg) as a white solid.

¹H-NMR(CD₃OD)：8.11-7.99(m，1H)，7.89-7.79(m，1H)，7.50-7.24(m，7H)，6.99-6.88(m，1H)，6.73-6.58(m，1H)，5.94-5.83(m，1H)，5.21-4.83(m，3H)，4.50-4.02(m，4H)，3.70-3.48(m，1H)，1.46-1.31(m，3H).

MS(ESI m/z)：705(M+H)

RT(min)：1.26

Examples 5 and 2

The same procedures as in example 5-1 were repeated to give the compounds of tables 3, 4-1 and 4-2.

[ Table 3]

[ Table 4-1]

[ tables 4-2]

Example 6-1

[ chemical formula 50]

First step of

(1) To a mixture of phosphorus oxychloride (96. mu.L) and dichloromethane (2.0mL) under a nitrogen atmosphere were added 4-chloro-3-fluorophenol (146mg) and triethylamine (150. mu.L) at-78 ℃ and the mixture was stirred for 1 hour. L-alanine benzyl ester hydrochloride (226mg) and triethylamine (300. mu.L) were added to the reaction mixture at-78 ℃ and the mixture was stirred at room temperature for 1.5 hours. The solvent was distilled off under reduced pressure, diethyl ether was added to the obtained residue, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give (2S) -benzyl 2- (((RS) -chloro (4-chloro-3-fluorophenoxy) phosphoryl) amino) propionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (100mg) and tetrahydrofuran (2.0mL) under a nitrogen atmosphere was added dropwise a 1.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (500. mu.L) at-78 ℃ and stirred for 30 minutes. A mixture of (2S) -benzyl 2- (((RS) -chloro (4-chloro-3-fluorophenoxy) phosphoryl) amino) propionate obtained in (1) and tetrahydrofuran (2.0mL) was added to the reaction mixture at-78 ℃ and stirred at room temperature for 12 hours. The reaction mixture was added with water, extracted with ethyl acetate, washed with a saturated aqueous solution of sodium chloride, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 100: 0 to 70: 30) to give (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-chloro-3-fluorophenoxy) phosphoryl) amino) propionate (79.7mg) as a colorless oil.

MS(ESI m/z)：931(M+H)

RT(min)：2.14

Second step of

To a mixture of (2S) -benzyl 2- (((RS) - ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (4-chloro-3-fluorophenoxy) phosphoryl) amino) propionate (79.7mg) and dichloromethane (1.0mL) was added trifluoroacetic acid (1.0mL), and the mixture was stirred at room temperature for 2.5 hours. The solvent was distilled off under reduced pressure, and methylene chloride (2.0mL) and triethylamine (1.0mL) were added to the obtained residue, and the solvent was distilled off under reduced pressure. The resulting residue was purified by basic silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 80: 20) to give (2S) -benzyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (4-chloro-3-fluorophenoxy) phosphoryl) amino) propionate (23.2mg) as a white solid.

¹H-NMR(CD₃OD)：8.09-7.99(m，1H)，7.48-7.40(m，1H)，7.39-7.29(m，5H)，7.23-7.12(m，1H)，7.10-7.01(m，1H)，6.71-6.61(m，1H)，5.92-5.86(m，1H)，5.19-5.04(m，3H)，4.45-4.19(m，3H)，4.14-3.97(m，1H)，3.63-3.51(m，1H)，1.43-1.34(m，3H).

MS(ESI m/z)：631(M+H)

RT(min)：1.32、1.33

Example 6 to 2

The same procedures as in example 6-1 were repeated to give the compounds shown in tables 5 and 6.

[ Table 5]

[ Table 6]

Examples 6 to 3

[ chemical formula 51]

First step of

The following compounds were obtained in the same manner as in the first step of example 6-1.

(2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (tert-Butoxycarbonylamino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-Butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (pyridin-3-oxy) phosphoryl) amino) propionate

MS(ESI m/z)：780(M+H)

RT(min)：1.59

Second step of

The following compounds were obtained in the same manner as in the second step of example 6-1.

(2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (pyridin-3-oxy) phosphoryl) amino) propionate

MS(ESI m/z)：580(M+H)

RT(min)：0.93

Example 7-1

[ chemical formula 52]

First step of

To a mixture of phosphorus oxychloride (84. mu.L) and dichloromethane (2.2mL) under a nitrogen atmosphere at-78 ℃ after addition of triethylamine (620. mu.L), a mixture of tert-butyl-tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (500mg) and dichloromethane (2.2mL) was added and stirred for 1 hour. A mixture of L-alanine benzyl ester hydrochloride (192mg) and methylene chloride (0.9mL) was added to the reaction mixture, which was then stirred at room temperature for 30 minutes. Water (0.5mL) was added, the mixture was stirred for 1.5 hours, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 95: 5 to 70: 30) to give (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) amino) propionate (308mg) as a yellow solid.

MS(ESI m/z)：803(M+H)

RT(min)：1.70

Second step of

Chloromethyl isopropyl carbonate (10. mu.L) was added to a mixture of (2S) -benzyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) amino) propionate (20.0mg), N-diisopropylethylamine (17. mu.L) and N, N-dimethylformamide (1.0mL), and stirred at 80 ℃ for 5 hours. Water was added to the reaction solution at room temperature, followed by extraction with isopropyl acetate. The organic layer was washed with water and a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. To the resulting residue were added dichloromethane (0.5mL) and trifluoroacetic acid (0.5mL), and the mixture was stirred at room temperature for 1.5 hours. The solvent was distilled off under reduced pressure, triethylamine (2.0mL) was added to the obtained residue, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 95: 5 to 88: 12) to give (2S) -benzyl 2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (((isopropoxycarbonyl) oxy) methoxy) phosphoryl) amino) propionate (6.0mg) as a white solid.

¹H-NMR(CD₃OD)：8.11-8.02(m，1H)，7.41-7.27(m，5H)，6.72-6.58(m，1H)，5.92(d，1H，J＝7.9Hz)，5.64-5.51(m，2H)，5.18(s，2H)，5.10-4.88(m，1H)，4.47-4.36(m，1H)，4.34-4.11(m，2H)，4.05-3.89(m，1H)，3.70-3.42(m，2H)，1.41(d，3H，J＝7.3Hz)，1.28(d，6H，J＝6.6Hz).

MS(ESI m/z)：619(M+H)

RT(min)：1.17

Example 7-2

[ chemical formula 53]

First step of

The following compounds were obtained in the same manner as in the first step of example 7-1.

(2S) -isopropyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) amino) propionate

MS(ESI m/z)：755(M+H)

RT(min)：1.61

Second step of

The following compounds were obtained in the same manner as in the second step of example 7-1.

(2S) -isopropyl 2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (((Isopropoxycarbonyl) oxy) methoxy) phosphoryl) amino) propanoate

¹H-NMR(CD₃OD)：8.14-8.01(m，1H)，6.74-6.58(m，1H)，5.97-5.88(m，1H)，5.69-5.54(m，2H)，5.11-4.78(m，3H)，4.50-4.40(m，1H)，4.38-4.15(m，2H)，3.94-3.77(m，1H)，3.65-3.50(m，1H)，1.44-1.34(m，3H)，1.33-1.16(m，12H).

MS(ESI m/z)：571(M+H)

RT(min)：1.05

Examples 7 to 3

The same procedure as in the second step of example 7-1 was repeated to give the compounds of Table 7.

[ Table 7]

Example 8

[ chemical formula 54]

First step of

To a mixture of tert-butyl-tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (100mg), 1H-tetrazole (49.9mg), and acetonitrile (0.5mL) was added a mixture of S, S' - (((((((((diisopropylamino) phosphinediyl) bis (oxy)) bis (ethane-2, 1-diyl)) bis (2, 2-dimethylthiopropionate) (161mg) and acetonitrile (0.5mL), and the mixture was stirred at room temperature for 21.5 hours. M-chloroperbenzoic acid (88.0mg) was added to the reaction mixture, and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture were added a saturated aqueous sodium bicarbonate solution (1.0mL) and sodium sulfite (10.0mg), and the mixture was stirred at room temperature for 0.5 hour and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 71: 29 to 50: 50) to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (S-pivaloyl-2-mercaptoethane-1-yl) phosphate (54.0mg) as a colorless oil.

MS(ESI m/z)：930(M+H)

RT(min)：2.26

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (S-pivaloyl-2-mercaptoethan-1-yl) phosphate (25.0mg) and dichloromethane (0.5mL) was added trifluoroacetic acid (0.5mL), and the mixture was allowed to stand at room temperature for 1 hour. The solvent was distilled off under reduced pressure, ethyl acetate (1.0mL) was added to the resulting residue with stirring, and a 4.0mol/L hydrogen chloride/1, 4-dioxane solution (10 μ L) was added. Hexane (0.5mL) was added to the reaction mixture, and after stirring for 1 hour, the precipitated solid was collected by filtration to give ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (S-pivaloyl-2-mercaptoethane-1-yl) phosphate hydrochloride as a white solid (12.0 mg).

¹H-NMR(DMSO-d₆)：9.36-9.15(m，1H)，8.39-8.25(m，1H)，8.19(d，1H，J＝7.6Hz)，6.42(dd，1H，J＝16.2，5.0Hz)，6.32-6.17(m，1H)，6.09(d，1H，J＝7.6Hz)，5.20-4.96(m，1H)，4.55-4.42(m，1H)，4.39-4.21(m，6H)，3.68-2.96(m，5H)，1.16(s，18H).

MS(ESI m/z)：630(M+H)

RT(min)：1.32

Example 9-1

[ chemical formula 55]

First step of

To a mixture of tert-butyl-tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (50.0mg), 1H-tetrazole (24.9mg) and acetonitrile (0.5mL) was added a mixture of S, S' - (((((((((diisopropylamino) phosphinediyl) bis (oxy)) bis (ethane-2, 1-diyl)) bis (2-methylthiopropionate) (75.8mg) and acetonitrile (0.4mL), and the mixture was stirred at room temperature for 18 hours. A mixture of iodine (57.4mg), water (90. mu.L) and pyridine (0.9mL) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour. Sodium sulfite (10.0mg) was added to the reaction solution, and the mixture was stirred at room temperature for 0.5 hour and then extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 70: 30 to 50: 50) to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (S-isobutyryl-2-mercaptoethan-1-yl) phosphate (63.3mg) as a colorless oil.

MS(ESI m/z)：902(M+H)

RT(min)：2.14

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (S-isobutyryl-2-mercaptoethan-1-yl) phosphate (61.3mg) and dichloromethane (0.5mL) was added trifluoroacetic acid (0.5mL), and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and to a mixture of the obtained residue and ethyl acetate (1.0mL), 4.0mol/L hydrogen chloride/1, 4-dioxane solution (26 μ L) was added, and stirred at room temperature for 30 minutes. Hexane (1.5mL) was added to the reaction mixture, the mixture was stirred for 1 hour, and the solvent was distilled off under reduced pressure. To the obtained residue was added ethyl acetate (1.0mL), and after stirring for 30 minutes, the precipitated solid was collected by filtration to give (hydrochloride of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (S-isobutyryl-2-mercaptoethane-1-yl) phosphate (13.7mg) as a white solid.

MS(ESI m/z)：602(M+H)

RT(min)：1.21

Example 9-2

The same procedures as in example 9-1 were repeated to give the compounds shown in tables 8 and 9.

[ Table 8]

[ Table 9]

Example 10-1

[ chemical formula 56]

First step of

(1) To a mixture of 2-cyanoethyl N, N-diisopropylchlorophosphamide (200mg), triethylamine (1.5mL) and tetrahydrofuran (2.0mL) was added a mixture of S- (2-hydroxyethyl) 2, 2-dimethylthiopropionate (137mg) and tetrahydrofuran (0.5mL) under ice-cooling under a nitrogen atmosphere, and the mixture was stirred at room temperature for 1.5 hours. After insoluble materials were filtered off, the solvent was distilled off under reduced pressure to obtain S- (2- (((2-cyanoethoxy) (diisopropylamino) phosphino) oxy) ethyl) 2, 2-dimethylthiopropionate.

(2) To a mixture of S- (2- (((2-cyanoethoxy) (diisopropylamino) phosphino) oxy) ethyl) 2, 2-dimethylthiopropionate obtained in (1), tert-butyl-tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (316mg) and tetrahydrofuran (3.0mL) was added 1H-tetrazole (118mg), and the mixture was stirred at room temperature for 2 hours. A mixture of iodine (214mg), water (0.1mL) and pyridine (1.0mL) was added to the reaction mixture, and the mixture was stirred for 30 minutes. Sodium sulfite (213mg) was added to the reaction solution, followed by extraction with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate (593 mg).

MS(ESI m/z)：839(M+H)

RT(min)：2.05

Second step of

A mixture of S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate (593mg) and 7.0mol/L ammonia/methanol solution (3.0mL) was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure to give an ammonium salt of S- (2- (((((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate (459 mg).

MS(ESI m/z)：786(M+H)

RT(min)：1.59

Third Process

A mixture of the ammonium salt of S- (2- ((((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate (50.0mg), chloromethylisopropyl carbonate (27. mu.L), N-diisopropylethylamine (43. mu.L) and N, N-dimethylformamide (0.5mL) was stirred at 80 ℃ for 2 hours. Water was added to the reaction solution at room temperature, followed by extraction with ethyl acetate. The organic layer was washed with water and a saturated aqueous sodium chloride solution, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2- (pivaloylthio) ethoxy) phosphoryl) oxy) methylpivalate.

MS(ESI m/z)：900(M+H)

RT(min)：2.21

The fourth step

To a mixture of (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2- (pivaloylthio) ethoxy) phosphoryl) oxy) methylpivalate and dichloromethane (0.5mL) was added trifluoroacetic acid (0.5mL), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, triethylamine (2.0mL) was added to the obtained residue, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol: 95: 5 to 88: 12) to give (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (2- (pivaloylthio) ethoxy) phosphoryl) oxy) methylpivalate (12.8mg) as a colorless oil.

¹H-NMR(CDCl₃)：8.06-7.95(m，1H)，6.76-6.60(m，1H)，6.10-5.98(m，1H)，5.73-5.58(m，2H)，5.27-5.01(m，1H)，4.63-4.51(m，1H)，4.42-4.05(m，4H)，3.75-3.64(m，1H)，3.21-3.03(m，3H)，1.23(s，18H).

MS(ESI m/z)：600(M+H)

RT(min)：1.35

Example 10-2

[ chemical formula 57]

First step of

The following compounds were obtained in the same manner as in the third step of example 10-1.

S- (2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (((isopropoxycarbonyl) oxy) methoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate

MS(ESI m/z)：902(M+H)

RT(min)：2.14

Second step of

The following compounds were obtained in the same manner as in the fourth step of example 10-1.

S- (2- (((RS) - (((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (((isopropoxycarbonyl) oxy) methoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate

¹H-NMR(CDCl₃)：8.03-7.92(m，1H)，6.85-6.72(m，1H)，5.99-5.89(m，1H)，5.72-5.60(m，2H)，5.27-5.03(m，1H)，5.01-4.86(m，1H)，4.66-4.52(m，1H)，4.42-4.07(m，4H)，3.75-3.64(m，1H)，3.21-3.08(m，2H)，1.32(d，6H，J＝6.6Hz)，1.24(s，9H).

MS(ESI m/z)：602(M+H)

RT(min)：1.22

Example 11-1

[ chemical formula 58]

First step of

To a mixture of phosphorus oxychloride (84. mu.L) and dichloromethane (2.2mL) under a nitrogen atmosphere at-78 ℃ after addition of triethylamine (620. mu.L), a mixture of tert-butyl-tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (500mg) and dichloromethane (2.2mL) was added and stirred for 1 hour. Water (0.5mL) was added to the reaction mixture, and the mixture was stirred at room temperature for 1 hour and then extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 95: 5 to 70: 30) to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methyl dihydrogenphosphate (306 mg).

MS(ESI m/z)：642(M+H)

RT(min)：1.33

Second step of

A mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methyl dihydrogenphosphate (100mg), chloromethylisopropyl carbonate (124. mu.L), N-diisopropylethylamine (221. mu.L) and N, N-dimethylformamide (1.0mL) was stirred at 60 ℃ for 4.5 hours and at 80 ℃ for 3 hours. Water was added to the reaction mixture at room temperature, extraction was performed with ethyl acetate, and the organic layer was washed with water and a saturated aqueous sodium chloride solution and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and trifluoroacetic acid (0.5mL) was added to a mixture of the obtained residue and methylene chloride (0.5mL), and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, triethylamine (2.0mL) was added to the obtained residue, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 100: 0 to 70: 30) to give ((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methoxy) (hydroxy) phosphoryl) oxy) methylisopropyl carbonate (2.8mg) and ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyl bis (isopropoxycarbonyloxymethyl) phosphate (1.8mg) as colorless oils.

Example 11-1

((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) oxy) methyl isopropyl carbonate

¹H-NMR(CD₃OD)：8.31-8.23(m，1H)，6.62-6.52(m，1H)，5.99-5.92(m，1H)，5.57-5.47(m，2H)，5.09-4.89(m，1H)，4.51-4.38(m，1H)，4.15-4.05(m，2H)，3.55-3.26(m，2H)，1.27(d，6H，J＝5.9Hz).

MS(ESI m/z)：458(M+H)

RT(min)：0.63

Example 11-1-2

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl bis (isopropoxycarbonyloxymethyl) phosphate

¹H-NMR(CDCl₃)：8.00(dd，1H，J＝7.6，1.7Hz)，6.84(dd，1H，J＝21.8，4.0Hz)，5.78(d，1H，J＝7.6Hz)，5.74-5.61(m，5H)，5.24-5.02(m，1H)，5.01-4.87(m，2H)，4.66-4.58(m，1H)，4.39-4.28(m，2H)，3.72-3.59(m，1H)，1.33(d，12H).

MS(ESI m/z)：574(M+H)

RT(min)：1.08

Example 11-2

The same procedure as in the second step of example 11-1 was repeated to give the compounds in Table 10.

[ Table 10]

Example 12-1

[ chemical formula 59]

First step of

To a mixture of phosphorus oxychloride (84 μ L) and dichloromethane (2.2mL) under a nitrogen atmosphere at-78 ℃ after addition of triethylamine (620 μ L) was added a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) -tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (500mg) and dichloromethane (2.2mL) was added and stirred for 1 hour. A mixture of 2, 2' -dithioalkanediyl diethanol (685mg) and methylene chloride (2.2mL) was added to the reaction mixture at-78 ℃ and the mixture was stirred for 1.5 hours and at room temperature for 3 hours. Water was added to the reaction mixture, which was stirred for 2 hours and extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 100: 0 to 70: 30) to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (2- ((2-hydroxyethyl) disulfide) ethyl) phosphate (218mg) as a yellow solid.

MS(ESI m/z)：914(M+H)

RT(min)：1.71

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (2- ((2-hydroxyethyl) disulfide) ethyl) phosphate (50.0mg) and dichloromethane (0.5mL), trifluoroacetic acid (0.5mL) was added and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, triethylamine (2.0mL) was added to the obtained residue, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol: 90: 10 to 70: 30) to obtain ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophene-2-yl) methyl bis (2- ((2-hydroxyethyl) disulfide) ethyl) phosphate (2.6mg) as a colorless oil.

¹H-NMR(CD₃OD)：8.08(dd，1H，J＝7.3，2.0Hz)，6.69(dd，1H，J＝19.5，4.3Hz)，5.94(d，1H，J＝7.3Hz)，5.12-4.89(m，1H)，4.51-4.26(m，7H)，3.79(t，4H，J＝6.3Hz)，3.68-3.59(m，1H)，3.04(t，4H，J＝6.3Hz)，2.87(t，4H，J＝6.3Hz).

MS(ESI m/z)：614(M+H)

RT(min)：0.80

Example 12-2

The same procedures as in example 12-1 were repeated to give the compounds in tables 11 and 12.

[ Table 11]

[ Table 12]

Example 13-1

[ chemical formula 60]

First step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (2- ((2-hydroxyethyl) disulfide) ethyl) phosphate (100mg), triethylamine (68.7 μ L) and tetrahydrofuran (1.0mL) was added pivaloyl chloride (36 μ L) under ice-cooling, and the mixture was stirred at room temperature for 2.5 hours. To the reaction mixture were added triethylamine (137. mu.L) and pivaloyl chloride (73. mu.L) under ice-cooling, and the mixture was stirred at room temperature for 5 hours, then added with a saturated aqueous sodium bicarbonate solution, stirred for 30 minutes, and then extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylbis (2- ((2-pivaloyloxyethyl) disulfide) ethyl) phosphate.

MS(ESI m/z)：1082(M+H)

RT(min)：2.33

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methyl bis (2- ((2-pivaloyloxyethyl) disulfide) ethyl) phosphate obtained in the first step and dichloromethane (1.0mL) was added trifluoroacetic acid (1.0mL), and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, triethylamine (2.0mL) was added to the obtained residue, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol: 94: 6 to 70: 30) to obtain ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophene-2-yl) methyl bis (2- ((2-pivaloyloxyethyl) disulfide) ethyl) phosphate (1.7mg) as a colorless oil.

¹H-NMR(CD₃OD)：8.07(dd，1H，J＝7.3，2.0Hz)，6.69(dd，1H，J＝19.5，4.3Hz)，5.93(d，1H，J＝7.3Hz)，5.11-4.90(m，1H)，4.51-4.26(m，11H)，3.70-3.59(m，1H)，3.05(t，4H，J＝5.9Hz)，2.99(t，4H，J＝6.3Hz)，1.20(s，18H).

MS(ESI m/z)：782(M+H)

RT(min)：1.67

Example 13-2

The same procedures as in example 13-1 were repeated to give the compounds shown in tables 13 and 14.

[ Table 13]

[ Table 14]

Example 14-1

[ chemical formula 61]

First step of

(1) To a mixture of phenyl dichlorophosphate (58. mu.L), triethylamine (99. mu.L) and dichloromethane (1.0mL) was added a mixture of S- (2-hydroxyethyl) 2, 2-dimethylthiopropionate (57.8mg) and dichloromethane (1.0mL) under ice-cooling under a nitrogen atmosphere, and the mixture was stirred under ice-cooling for 2 hours. After the solvent was distilled off under reduced pressure, methyl t-butyl ether (5.0mL) was added and insoluble matter was filtered off. The solvent was distilled off under reduced pressure to give S- (2- (((RS) -chloro (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate.

(2) To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) -tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (100mg) and tetrahydrofuran (1.0mL) under a nitrogen atmosphere was added 1.0mol/L of a tert-butylmagnesium chloride/tetrahydrofuran solution (214. mu.L) at-78 ℃ and stirred for 30 minutes. A mixture of S- (2- ((chloro (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate obtained in (1) and tetrahydrofuran (1.0mL) was added to the reaction mixture, and the mixture was stirred at room temperature for 42 hours. Water was added to the reaction solution, extraction was performed with ethyl acetate, and the organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate as a crude product.

MS(ESI m/z)：862(M+H)

RT(min)：2.18

Second step of

Trifluoroacetic acid (0.5mL) was added to a mixture of S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate obtained in the first step and dichloromethane (0.5mL), and the mixture was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and a saturated aqueous sodium hydrogencarbonate solution was added to the obtained residue to conduct extraction with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform: methanol ═ 100: 0 to 70: 30) to give S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate (1.7mg) as a colorless oil.

¹H-NMR(CD₃OD)：8.08-7.99(m，1H)，7.46-7.21(m，5H)，6.73-6.62(m，1H)，5.89(d，1H，J＝7.3Hz)，5.37-5.04(m，1H)，4.55-4.32(m，3H)，4.31-4.20(m，2H)，3.68-3.58(m，1H)，3.19(t，2H，J＝6.3Hz)，1.24-1.15(m，9H).

MS(ESI m/z)：562(M+H)

RT(min)：1.28

Example 14-2

[ chemical formula 62]

First step of

The following compounds were obtained in the same manner as in the first step of example 14-1.

S- (4- (((RS) - (((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (phenoxy) phosphoryl) oxy) butyl) 2, 2-dimethylthiopropionate

MS(ESI m/z)：890(M+H)

RT(min)：2.24

Second step of

The following compounds were obtained in the same manner as in the second step of example 14-1.

S- (4- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (phenoxy) phosphoryl) oxy) butyl) 2, 2-dimethylthiopropionate

¹H-NMR(CD₃OD)：8.08-7.99(m，1H)，7.46-7.19(m，5H)，6.73-6.61(m，1H)，5.89(d，1H，J＝7.9Hz)，5.10-4.96(m，1H)，4.53-4.31(m，3H)，4.29-4.19(m，2H)，3.68-3.58(m，1H)，2.86(t，2H，J＝6.9Hz)，1.84-1.55(m，4H)，1.21(s，9H).

MS(ESI m/z)：590(M+H)

RT(min)：1.38

Example 15-1

[ chemical formula 63]

To a mixture of L-phenylalanine ethyl ester hydrochloride (644mg), phosphorus oxychloride (130. mu.L) and methylene chloride (5.6mL) was added triethylamine (775. mu.L) at-78 ℃ and the mixture was stirred at room temperature for 25 minutes. To the reaction solution were added methanesulfonate (100mg) and triethylamine (775. mu.L) of 4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one, and after stirring at room temperature for 3 hours, a saturated aqueous ammonium chloride solution was added, followed by extraction with chloroform and drying over magnesium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol: 100: 0 to 85: 15) to obtain a colorless oil. The obtained colorless oil was purified by reverse phase fractional HPLC (0.1% aqueous formic acid solution-0.1% acetonitrile formic acid solution) to give a formate salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyl N, N' -bis ((1S) -1-ethoxycarbonyl-2-phenylethane-1-yl) phosphorodiamidate as a colorless oil (3.2 mg).

¹H-NMR(CD₃OD)：8.32(d，1H，J＝7.6Hz)，8.12(s，1H)，7.28-7.04(m，10H)，6.62-6.49(m，1H)，5.92(d，1H，J＝7.6Hz)，5.19-4.93(m，1H)，4.50-4.37(m，1H)，4.22-4.07(m，4H)，4.01-3.90(m，2H)，3.87-3.80(m，2H)，3.44-3.33(m，1H)，3.27-3.13(m，4H)，1.39-1.18(m，3H)，1.14-1.02(m，3H).

MS(ESI m/z)：692(M+H)

RT(min)：1.27

Example 15-2

[ chemical formula 64]

The following compounds were obtained in the same manner as in example 15-1.

Formate salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl N, N' -bis ((1S) -1-ethoxycarbonyl-3-methylbutan-1-yl) phosphorodiamidite

¹H-NMR(CD₃OD)：8.55-8.42(m，1H)，6.61-6.48(m，1H)，6.24-6.11(m，1H)，5.13-4.90(m，1H)，4.45-4.31(m，1H)，4.17(q，2H，J＝7.0Hz)，4.02(q，2H，J＝7.3Hz)，3.96-3.76(m，4H)，3.42-3.32(m，1H)，1.92-1.75(m，1H)，1.75-1.60(m，1H)，1.60-1.45(m，4H)，1.35-1.09(m，6H)，1.01-0.91(m，6H)，0.91-0.76(m，6H).

MS(ESI m/z)：624(M+H)

RT(min)：1.27

Example 16-1

[ chemical formula 65]

First step of

To a mixture of phosphorus oxychloride (17 μ L) and dichloromethane (0.9mL) under a nitrogen atmosphere at-78 ℃ after addition of triethylamine (248 μ L) was added a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (50.0mg) and dichloromethane (0.9mL) was added and stirred for 30 minutes. A mixture of L-alanine benzyl ester hydrochloride (76.8mg) and methylene chloride (1.7mL) was added to the reaction mixture at-78 ℃ and the mixture was stirred for 20 minutes and then at room temperature for 30 minutes. Water was added to the reaction solution, extraction was performed with ethyl acetate, and the organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methyl N, N' -bis ((1S) -1-benzyloxycarbonylethan-1-yl) phosphorodiamidate.

MS(ESI m/z)：964(M+H)

RT(min)：2.05

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methyl N, N' -bis ((1S) -1-benzyloxycarbonylethan-1-yl) phosphorodiamidate and dichloromethane (0.5mL) was added trifluoroacetic acid (0.5mL), and the mixture was stirred at room temperature for 1.5 hours. The solvent was distilled off under reduced pressure, and a saturated aqueous sodium hydrogencarbonate solution was added to the obtained residue to conduct extraction with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by basic silica gel column chromatography (chloroform: methanol ═ 94: 6 to 86: 14) to give ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl N, N' -bis ((1S) -1-benzyloxycarbonylethane-1-yl) phosphorodiamidite (6.0mg) as a white solid.

¹H-NMR(DMSO-d₆)：7.88-7.82(m，1H)，7.39-7.18(m，12H)，6.53(dd，1H，J＝17.8，4.6Hz)，6.01(d，1H，J＝4.6Hz)，5.79(d，1H，J＝7.3Hz)，5.16-4.81(m，7H)，4.36-4.25(m，1H)，4.18-4.05(m，1H)，3.99-3.76(m，3H)，3.52-3.24(m，1H)，1.33-1.21(m，6H).

MS(ESI m/z)：664(M+H)

RT(min)：1.21

Example 16-2

The same procedures as in example 16-1 were repeated to give the compounds shown in tables 15 and 16.

[ Table 15]

[ Table 16]

Example 17

[ chemical formula 66]

To a mixture of the mesylate salt of 4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (50.0mg), diphenyl chlorophosphate (35. mu.L), and pyridine (1.0mL) was added N, N-dimethylaminopyridine (1.7 mg). After stirring at room temperature for 1 hour, diphenyl chlorophosphate (35. mu.L) was added to the reaction mixture, and the mixture was stirred at room temperature for 2 hours. Methanol (35. mu.L) was added to the reaction solution, and the mixture was purified by silica gel column chromatography (ethyl acetate: methanol: 90: 10 to 81: 19) to obtain a white solid. The resulting white solid was purified by reverse phase fractional HPLC (0.1% aqueous formic acid-0.1% formic acid in acetonitrile) to give the formate salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyldiphenylphosphate (2.0mg) as a white solid.

¹H-NMR(CDCl₃)：8.12(s，1H)，7.89-7.71(m，1H)，7.39-7.10(m，12H)，7.06-6.93(m，1H)，6.68-6.49(m，1H)，5.94-5.77(m，1H)，5.19-4.88(m，1H)，4.50-4.22(m，3H)，3.72-3.59(m，1H).

MS(ESI m/z)：494(M+H)

RT(min)：1.08

Example 18

[ chemical formula 67]

To a suspension of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((di-t-butoxyphosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (50.0mg) in methanol (1.0mL) was added 28% sodium methoxide/methanol solution (15.0mL), and the mixture was stirred at room temperature for 1.5 hours. Acetic acid (30 μ L) was added to the reaction solution, the solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 70: 30) to obtain ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophene-2-yl) methyl di-tert-butyl phosphate (28.8mg) as a white solid.

¹H-NMR(DMSO-d₆)：7.92-7.87(m，1H)，7.33-7.21(m，2H)，6.53(dd，1H，J＝17.1，5.1Hz)，6.06fd，1H，J＝4.8Hz)，5.78(d，1H，J＝7.2Hz)，5.08-4.86(m，1H)，4.36-4.25(m，1H)，4.25-4.14(m，1H)，4.10-3.98(m，1H)，3.48-3.38(m，1H)，1.43(s，18H).

MS(ESI m/z)：454(M+H)

RT(min)：0.93

Example 19-1

[ chemical formula 68]

A mixture of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl dihydrogenphosphate (100mg), 2- ((ethylcarbamoyl) thio) ethyl 4-methylbenzenesulfonate (445mg), N-diisopropylethylamine (250. mu.L) and N, N-dimethylformamide (1.0mL) was stirred at 80 ℃ for 6 hours. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (chloroform: methanol: 86: 14 to 39: 61) to give S- (2- (((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) oxy) ethyl) ethylthiocarbamate (2.9mg) as a colorless oil.

¹H-NMR(CD₃OD)：8.30(d，1H，J＝7.3Hz)，6.56(dd，1H，J＝13.2，5.3Hz)，5.98(d，1H，J＝7.9Hz)，5.11-4.87(m，1H)，4.49-4.38(m，1H)，4.19-4.07(m，2H)，4.04-3.91(m，2H)，3.57-3.41(m，1H)，3.27-3.07(m，4H)，1.10(t，3H，J＝7.3Hz).

MS(ESI m/z)：473(M+H)

RT(min)：0.56

Example 19-2

The same procedures as in example 19-1 were repeated to give the compounds shown in Table 17.

[ Table 17]

Example 20-1

[ chemical formula 69]

First step of

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- (hydroxymethyl) tetrahydrothiophene-3-benzoic acid ester (100mg), 2-cyanoethyloctadecyldiisopropylphosphoramidite (314mg), 1H-tetrazole (100mg), and dichloromethane (4.0mL) was stirred at room temperature for 5 hours. After standing for 14.5 hours, m-chloroperbenzoic acid (60.0mg) was added at-78 ℃ and the mixture was heated to-10 ℃ over 1.5 hours and stirred for 1 hour. Water was added to the reaction mixture, followed by extraction with dichloromethane, and the organic layer was washed with a saturated aqueous sodium bicarbonate solution, a 10% aqueous sodium bisulfite solution, and water, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate: 57: 43 to 0: 100) to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((RS) - (2-cyanoethoxy) (octadecyloxy) phosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester as a colorless oil (41.0 mg).

MS(ESI m/z)：855(M+H)

RT(min)：2.64

Second step of

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- ((((RS) - (2-cyanoethoxy) (octadecyloxy) phosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester (41.0mg) and 7.0mol/L ammonia/methanol solution (4.0mL) was stirred at room temperature for 10 hours. After standing for 12 hours, the solvent was distilled off under reduced pressure, and acetone was added to the obtained residue to collect the precipitated solid by filtration, thereby obtaining an ammonium salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophene-2-yl) methyloctadecylhydrogenphosphate (13.8mg) as a white solid.

¹H-NMR(DMSO-d₆)：7.99(d，1H，J＝7.3Hz)，7.43-6.89(m，4H)，6.44(dd，1H，J＝12.9，5.6Hz)，5.79(d，1H，J＝7.3Hz)，5.06-4.78(m，1H)，4.32-4.20(m，1H)，4.01-3.78(m，2H)，3.71-3.57(m，2H)，3.50-3.17(m，1H)，1.56-1.37(m，2H)，1.37-1.12(m，30H)，0.92-0.78(m，3H).

MS(ESI m/z)：594(M+H)

RT(min)：2.12

Example 20-2

The same procedures as in example 20-1 were repeated to give the compounds shown in tables 18 and 19.

[ Table 18]

[ Table 19]

Example 21-1

[ chemical formula 70]

To a mixture of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methyl dihydrogenphosphate (100mg), L-alanine benzyl ester hydrochloride (126mg), triethylamine (82. mu.L), tert-butyl alcohol (4.0mL) and water (1.0mL) was added a mixture of N, N' -dicyclohexylcarbodiimide (90.0mg) and tert-butyl alcohol (1.0mL), and the mixture was stirred under reflux with heating for 4 hours. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol ═ 50: 50 to 25: 75) to give (2S) -benzyl 2- ((((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) amino) propionate as a white solid (62.3 mg).

¹H-NMR(CD₃OD)：8.30(d，1H，J＝7.3Hz)，7.46-7.22(m，5H)，6.60-6.43(m，1H)，5.97(d，1H，J＝7.3Hz)，5.34-4.99(m，3H)，4.51-4.31(m，1H)，4.18-3.87(m，3H)，3.66-3.50(m，1H)，1.46(d，3H，J＝6.6Hz).

MS(ESI m/z)：503(M+H)

RT(min)：0.79

Example 21-2

The same procedures as in example 21-1 were repeated to give the compounds in Table 20.

[ Table 20]

Example 22

[ chemical formula 71]

A mixture of (2S) -methyl 2- (((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) amino) propionate (10.0mg), water (100. mu.L) and triethylamine (100. mu.L) was stirred at room temperature for 11.5 hours. The solvent was distilled off under reduced pressure, and acetone was added to the obtained residue to collect the precipitated solid by filtration, thereby obtaining a triethylamine salt of (2S) -2- ((((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) amino) propanealkanoic acid as a white solid (4.3 mg).

MS(ESI m/z)：413(M+H)

RT(min)：0.39

Example 23-1

[ chemical formula 72]

First step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylhydrogenphosphonate (214mg), 2-aminopyridine (64.0mg), pyridine (340. mu.L) and dichloromethane (3.4mL) was added diphenyl chlorophosphate (142. mu.L), and the mixture was stirred at room temperature for 10 minutes. A mixture of iodine (96.0mg), pyridine (3.4mL) and water (308. mu.L) was added, and the mixture was stirred at room temperature for 30 minutes. After addition of an aqueous sodium hydrogen sulfite solution, the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (0.5% triethylamine in chloroform: methanol: 100: 0 to 50: 50) to obtain a colorless oil. The resulting colorless oil was purified by silica gel column chromatography (0.5% triethylamine/chloroform: methanol 100: 0 to 70: 30) to give a triethylamine salt of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylhydrogen (pyridin-2-yl) phosphoramidate (213mg) as a colorless oil.

MS(ESI m/z)：718(M+H)

RT(min)：1.40

Second step of

To a mixture of ((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-2-yl) methylhydrogen (pyridin-2-yl) phosphoramidate (213mg) and dichloromethane (2.0mL) was added trifluoroacetic acid (2.0mL), and the mixture was stirred at room temperature for 30 minutes. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (0.5% triethylamine/chloroform: methanol: 100: 0 to 0: 100) to obtain a triethylamine salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methylhydropyridin-2-ylaminophosphate as a white solid (29.5 mg).

¹H-NMR(DMSO-d₆)：8.05-7.96(m，1H)，7.92-7.84(m，1H)，7.54-7.44(m，1H)，7.39-7.14(m，3H)，6.96-6.79(m，1H)，6.72-6.61(m，1H)，6.49-6.36(m，1H)，6.33-6.15(m，1H)，5.79-5.70(m，1H)，5.01-4.73(m，1H)，4.31-4.16(m，1H)，4.04-3.75(m，2H)，3.38-3.25(m，1H)，3.07-2.93(m，6H)，1.22-1.12(m，9H).

MS(ESI m/z)：418(M+H)

RT(min)：0.31

Example 23-2

[ chemical formula 73]

First step of

The following compounds were obtained in the same manner as in the first step of example 23-1.

((2R, 3S, 4S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4-fluorotetrahydrothiophen-3-yl) methylhydrogen (pyridin-2-yl) phosphoramidate

MS(ESI m/z)：718(M+H)

RT(min)：1.29

Second step of

The following compounds were obtained in the same manner as in the second step of example 23-1.

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methylhydrogen (pyridin-3-yl) phosphoramidate

¹H-NMR(DMSO-d₆)：8.29-8.22(m，1H)，7.90-7.84(m，1H)，7.82-7.78(m，1H)，7.47-7.40(m，1H)，7.34-7.14(m，2H)，7.05-6.99(m，1H)，6.79-6.71(m，1H)，6.48-6.35(m，1H)，6.34-6.23(m，1H)，5.80-5.71(m，1H)，5.05-4.75(m，1H)，4.29-4.15(m，1H)，3.96-3.73(m，2H)，3.31-3.24(m，1H)，3.12-2.91(m，6H)，1.24-1.09(m，9H).

MS(ESI m/z)：418(M+H)

RT(min)：0.26

Example 24-1

[ chemical formula 74]

First step of

To a mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- ((((2-cyanoethoxy) (diisopropylamino) phosphino) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoate (203mg), (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2-hydroxyethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (79.0mg) and acetonitrile (4.0mL) was added 1H-tetrazole (28.0mg), and the mixture was stirred at room temperature for 35 minutes. 1H-tetrazole (28.0mg) was added to the reaction mixture, and the mixture was stirred at room temperature for 5 minutes. A mixture of iodine (103mg), pyridine (2.0mL) and water (200. mu.L) was added to the reaction mixture, and the mixture was stirred at room temperature for 10 minutes. Adding an aqueous solution of sodium bisulfite to the reaction mixture, distilling off the solvent under reduced pressure, purifying the obtained residue by silica gel column chromatography (ethyl acetate: methanol: 100: 0 to 90: 10), (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- (((((((2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -3- (benzoyloxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (49.0mg) was obtained as a colorless oil.

MS(ESI m/z)：977(M+H)

RT(min)：1.45

Second step of

A mixture of (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((((((2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -3- (benzoyloxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (49.0mg) and 7.0mol/L ammonia/methanol solution (3.0mL) was stirred at room temperature for 8 hours 40 minutes. The solvent was distilled off under reduced pressure, water was added to the obtained residue, and the aqueous layer was washed 3 times with ethyl acetate. The solvent was distilled off under reduced pressure to give ammonium salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyl (2- (((2R, 3R, 4S, 5S, 6R) -3, 4, 5-trihydroxy-6- (hydroxymethyl) tetrahydro-2H-pyran-2-yl) oxy) ethylhydrogen phosphate (27.3mg) as a white solid.

¹H-NMR(D₂O)：8.29(d，1H，J＝7.9Hz)，6.34-6.28(m，1H)，6.01(d，1H，J＝7.9Hz)，5.18-4.93(m，1H)，4.41-4.30(m，2H)，4.09-3.95(m，5H)，3.81-3.76(m，2H)，3.62-3.54(m，1H)，3.44-3.16(m，5H).

MS(ESI m/z)：548(M+H)

RT(min)：0.26

Example 24-2

[ chemical formula 75]

First step of

To a mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- ((((2-cyanoethoxy) (diisopropylamino) phosphino) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoate (68.0mg), (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2-hydroxyethyl) disulfide) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (33.0mg) and acetonitrile (1.4mL) was added 1H-tetrazole (9.5mg), and the mixture was stirred at room temperature for 15 minutes. A mixture of iodine (35.0mg), pyridine (680. mu.L) and water (68. mu.L) was added to the reaction mixture, and the mixture was stirred at room temperature for 10 minutes. The solvent was distilled off under reduced pressure, the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol: 100: 0 to 90: 10), (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2- ((((((2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -3- (benzoyloxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethyl) dithio) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (36.0mg) was obtained as a colorless oil.

MS(ESI m/z)：1069(M+H)

RT(min)：1.56

Second step of

A mixture of (2R, 3R, 4S, 5R, 6R) -2- (acetoxymethyl) -6- (2- ((2- ((((((2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -3- (benzoyloxy) -4-fluorotetrahydrothiophen-2-yl) methoxy) (2-cyanoethoxy) phosphoryl) oxy) ethyl) disulfide) ethoxy) tetrahydro-2H-pyran-3, 4, 5-triyltriacetate (36.0mg) and 7.0mol/L ammonia/methanol solution (3.0mL) was stirred at room temperature for 8 hours 40 minutes. The solvent was distilled off under reduced pressure, water was added to the obtained residue, and the aqueous layer was washed 3 times with ethyl acetate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by reverse phase fractionation HPLC (0.1% aqueous formic acid solution-0.1% formic acid acetonitrile solution) to give a formate salt of ((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothiophen-2-yl) methyl (2- ((2- (((2R, 3R, 4S, 5S, 6R) -3, 4, 5-trihydroxy-6- (hydroxymethyl) tetrahydro-2H-pyran-2-yl) oxy) ethyl) disulfide) ethyl) hydrogen phosphate (7.6mg) as a white solid.

¹H-NMR(D₂O)：8.52(d，1H，J＝7.9Hz)，6.31-6.25(m，1H)，6.16(d，1H，J＝7.9Hz)，5.23-4.99(m，1H)，4.41-4.29(m，2H)，4.13-4.00(m，5H)，3.91-3.76(m，2H)，3.65-3.56(m，1H)，3.47-3.12(m，5H)，2.95-2.84(m，4H).

MS(ESI m/z)：640(M+H)

RT(min)：0.51

Example 25

[ chemical formula 76]

First step of

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-2- ((phosphonooxy) methyl) tetrahydrothiophene-3-benzoic acid ester (43.3mg), 1-chloroethyldiethylcarbamate (280mg), N-diisopropylethylamine (0.60mL), and N, N-methylformamide (0.9mL) was stirred at 50 ℃ for 3 hours. Water was added to the reaction mixture at room temperature, followed by extraction with ethyl acetate, and the organic layer was washed with water and a saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to give (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((1- ((diethylcarbamoyl) oxy) ethoxy) (hydroxy) phosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoic acid ester.

MS(ESI m/z)：693(M+H)

RT(min)：1.24

Second step of

A mixture of (2R, 3S, 4S, 5R) -5- (4-benzamide-2-oxopyrimidin-1 (2H) -yl) -2- (((1- ((diethylcarbamoyl) oxy) ethoxy) (hydroxy) phosphoryl) oxy) methyl) -4-fluorotetrahydrothiophene-3-benzoate obtained in the first step and 7.0mol/L ammonia/methanol solution (1.5mL) was stirred at room temperature for 16.5 hours. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 20: 80) to obtain 1- (((((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4-fluoro-3-hydroxytetrahydrothien-2-yl) methoxy) (hydroxy) phosphoryl) oxy) ethyl diethyl carbamate (21.7mg) as a yellow solid.

¹H-NMR(CD₃OD)：8.40-8.19(m，1H)，6.63-6.46(m，1H)，6.46-6.32(m，1H)，6.14-5.99(m，1H)，5.13-4.88(m，1H)，4.48-4.34(m，1H)，4.22-4.05(m，2H)，3.56-3.43(m，1H)，3.43-3.23(m，4H)，1.53(d，3H，J＝5.3Hz)，1.36-1.27(m，6H)

MS(ESI m/z)：485(M+H)

RT(min)：0.72

Example 26-1

[ chemical formula 77]

First step of

To a mixture of tert-butyl tert-butoxycarbonyl (1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate (168mg) and tetrahydrofuran (17mL) under a nitrogen atmosphere was added dropwise a 2.0mol/L tert-butylmagnesium chloride/tetrahydrofuran solution (450. mu.L) at-78 ℃ and stirred for 15 minutes. A mixture of 2- (4-nitrophenoxy) -1, 3, 2-dioxaphosphorinane 2-oxide (389mg) and tetrahydrofuran (3.0mL) is added to the reaction mixture at-78 deg.C, and the mixture is stirred at room temperature for 1.5 hours. To the reaction mixture was added a saturated aqueous ammonium chloride solution, followed by extraction with ethyl acetate, washing with a saturated aqueous sodium chloride solution, and drying over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate ═ 20: 80 to 80: 20) to give 4- (bis (tert-butoxycarbonyl) amino) -1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (((2-oxide-1, 3, 2-dioxaphosphorinan-2-yl) oxy) methyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (204mg) as a brown oil.

MS(ESI m/z)：682(M+H)

RT(min)：1.70

Second step of

A mixture of 4- (bis (tert-butoxycarbonyl) amino) -1- ((2R, 3S, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3-fluoro-5- (((2-oxide-1, 3, 2-dioxaphosphorinan-2-yl) oxy) methyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (204mg) and trifluoroacetic acid (1.0mL) was stirred at room temperature for 1.5 hours. The solvent was distilled off under reduced pressure, and the obtained residue was purified by silica gel column chromatography (ethyl acetate: methanol ═ 100: 0 to 80: 20) to give 4-amino-1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (((2-oxide-1, 3, 2-dioxaphosphorinan-2-yl) oxy) methyl) tetrahydrothiophen-2-yl) pyrimidin-2 (1H) -one (47.0mg) as a white solid.

¹H-NMR(DMSO-d₆)：7.90-7.88(m，1H)，7.34(s，1H)，7.28(s，1H)，6.59-6.53(m，1H)，6.12(d，1H，J＝4.6Hz)，5.81-5.78(m，1H)，5.10-4.85(m，1H)，4.45-4.31(m，6H)，4.22-4.11(m，1H)，3.56-3.47(m，1H)，2.14-1.79(m，2H).

MS(ESI m/z)：382(M+H)

RT(min)：0.49

Example 26-2

The same procedures as in example 26-1 were repeated to give the compounds shown in tables 21 and 22.

[ Table 21]

[ Table 22]

Example 27

[ chemical formula 78]

First step of

A mixture of N- (1- ((2R, 3S, 4S, 5R) -3-fluoro-4-hydroxy-5- (hydroxymethyl) tetrahydrothiophen-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (920mg), 2-cyanoethyl N, N, N ', N' -tetraisopropylphosphorodiamidite (800mg), 1H-tetrazole (530mg) and dichloromethane (10.0mL) was stirred at room temperature for 1 hour 20 minutes. A mixture of m-chloroperbenzoic acid (618mg) and methylene chloride (10.0mL) was added to the reaction mixture at-40 ℃ and the mixture was stirred for 2 hours. To the reaction solution were added a saturated aqueous sodium bicarbonate solution and a 10% aqueous sodium bisulfite solution, followed by extraction with dichloromethane. The organic layer was washed with water and a saturated aqueous sodium chloride solution, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and a mixture of hexane and ethyl acetate (═ 1/1) was added to the obtained residue, and the precipitated solid was collected by filtration to give N- (1- ((2RS, 4aR, 6R, 7S, 7aS) -2- (2-cyanoethoxy) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphin-6-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (580mg) aS a white solid.

MS(ESI m/z)：481(M+H)

RT(min)：1.02，1.05

Second step of

A mixture of N- (1- ((2RS, 4aR, 6R, 7S, 7aS) -2- (2-cyanoethoxy) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphin-6-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (580mg) and 7.0mol/L ammonia/methanol solution (10.0mL) was stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, and ethyl acetate was added to the obtained residue to collect a precipitated solid by filtration, thereby obtaining N- (1- ((4aR, 6R, 7S, 7aS) -7-fluoro-2-hydroxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-6-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (467mg) aS a pale yellow solid.

MS(ESI m/z)：428(M+H)

RT(min)：0.74

Third Process

A mixture of N- (1- ((4aR, 6R, 7S, 7aS) -7-fluoro-2-hydroxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-6-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) benzamide (350mg) and 7.0mol/L ammonia/methanol solution (4.0mL) was stirred at 40 ℃ for 4 hours. The solvent was distilled off under reduced pressure, ethyl acetate was added to the obtained residue, and the precipitated solid was collected by filtration to give 4-amino-1- ((4aR, 6R, 7S, 7aS) -7-fluoro-2-hydroxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-6-yl) pyrimidin-2 (1H) -one (265mg) aS a pale yellow solid.

¹H-NMR(CD₃OD)：8.05(d，1H，J＝7.9Hz)，6.72-6.60(m，1H)，5.98(d，1H，J＝7.9Hz)，5.33-5.04(m，1H)，4.83-4.65(m，1H)，4.41-4.21(m，2H)，3.57-3.39(m，1H).

MS(ESI m/z)：324(M+H)

RT(min)：0.21，0.26

Example 28

[ chemical formula 79]

To a mixture of 4-amino-1- ((4aR, 6R, 7S, 7aS) -7-fluoro-2-hydroxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphin-6-yl) pyrimidin-2 (1H) -one (20.0mg), trimethyl phosphate (1.0mL), N-dimethylformamide (10. mu.L) and dichloromethane (1.0mL) was added oxalyl chloride (20. mu.L) under ice-cooling, and the mixture was stirred for 40 minutes. Isopropyl alcohol (2.0mL) was added to the reaction mixture, and the mixture was stirred at room temperature for 3 hours and 40 minutes and then allowed to stand for 12 hours. The solvent was distilled off under reduced pressure, methylene chloride was added to the obtained residue, and insoluble matter was filtered off. The solvent was distilled off under reduced pressure, and water was added to the obtained residue, which was extracted with dichloromethane and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, diisopropyl ether was added to the obtained residue, and the precipitated solid was collected by filtration to give 4-amino-1- ((2RS, 4aR, 6R, 7S, 7aS) -7-fluoro-2-isopropoxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-6-yl) pyrimidin-2 (1H) -one (3.4mg) aS a pale yellow solid.

¹H-NMR(CD₃OD)：8.08-8.02(m，1H)，6.75-6.68(m，1H)，5.97(d，1H，J＝7.3Hz)，5.47-5.19(m，1H)，5.14-4.99(m，1H)，4.82-4.70(m，1H)，4.70-4.56(m，2H)，3.84-3.71(m，1H)，1.37(d，6H，J＝5.9Hz).

MS(ESI m/z)：366(M+H)

RT(min)：0.71

Example 29-1

[ chemical formula 80]

To a mixture of 4-amino-1- ((4aR, 6R, 7S, 7aS) -7-fluoro-2-hydroxy-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphin-6-yl) pyrimidin-2 (1H) -one (20.0mg), trimethyl phosphate (1.0mL), N-dimethylformamide (10. mu.L) and dichloromethane (1.0mL) was added oxalyl chloride (40. mu.L) under ice-cooling, and the mixture was stirred at room temperature for 1 hour and 20 minutes. To a mixture of L-alanine benzyl ester hydrochloride (20.0mg), N-diisopropylethylamine (200. mu.L) and dichloromethane (2.0mL) was added the reaction mixture under ice-cooling, and the mixture was stirred at room temperature for 2 hours and 40 minutes. Water was added to the reaction solution, which was extracted with dichloromethane and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, diisopropyl ether was added to the obtained residue, and the precipitated solid was collected by filtration to give (2S) -benzyl 2- (((2RS, 4aR, 6R, 7S, 7aS) -6- (4-amino-2-oxopyrimidin-1 (2H) -yl) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-2-yl) amino) propionate (6.7mg) aS a pale brown solid.

¹H-NMR(CD₃OD)：8.07-8.01(m，1H)，7.41-7.30(m，5H)，6.73-6.66(m，1H)，5.97(d，1H，J＝7.3Hz)，5.40-5.11(m，3H)，5.02-4.91(m，1H)，4.63-4.47(m，2H)，4.04-3.89(m，1H)，3.65-3.50(m，1H)，1.39(d，3H，J＝5.9Hz).

MS(ESI m/z)：485(M+H)

RT(min)：1.00

Example 29-2

[ chemical formula 81]

The following compounds were obtained in the same manner as in example 29-1.

(2S) -isopropyl 2- (((2RS, 4aR, 6R, 7S, 7aS) -6- (4-amino-2-oxopyrimidin-1 (2H) -yl) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-2-yl) amino) propionate

¹H-NMR(CD₃OD)：8.05(d，1H，J＝7.6Hz)，6.74-6.67(m，1H)，5.97(d，1H，J＝7.6Hz)，5.42-5.15(m，1H)，5.06-4.91(m，2H)，4.65-4.52(m，2H)，3.95-3.81(m，1H)，3.70-3.56(m，1H)，1.41-1.32(m，3H)，1.32-1.20(m，6H).

MS(ESI m/z)：437(M+H)

RT(min)：0.81

Comparative example 1

The following compounds were obtained according to the methods described in Journal of medicinal Chemistry, volume 52, pages 1531 to 1542, and year 2014.

[ chemical formula 82]

(2S) -benzyl 2- (((RS) - ((((2R, 3R, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4, 4-difluoro-3-hydroxytetrahydrofuran-2-yl) methoxy) (phenoxy) phosphoryl) amino) propionate

¹H-NMR(CD₃OD)：7.57-7.45(m，1H)，7.39-7.26(m，6H)，7.26-7.14(m，3H)，6.28-6.17(m，1H)，5.89-5.78(m，1H)，5.19-5.07(m，2H)，4.50-4.38(m，1H)，4.38-4.26(m，1H)，4.26-4.10(m，1H)，4.10-3.94(m，2H)，1.39-1.32(m，3H).

MS(ESI m/z)：581(M+H)

RT(min)：1.14

Comparative example 2

The following compounds were obtained according to the methods described in Nucleotides, Nucleotides & Nucleic Acids, Vol.24, Nos. 10 to 12, pp.1635 to 1649, and 2005.

[ chemical formula 83]

((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -3, 4-dihydroxytetrahydrofuran-2-yl) methyl bis (S-pivaloyl-2-mercaptoethane-1-yl) phosphate

¹H-NMR(CD₃OD)：7.81(d，1H，J＝7.3Hz)，6.26-6.20(m，1H)，5.88(d，1H，J＝7.3Hz)，4.46-4.23(m，2H)，4.21-4.07(m，6H)，4.04-3.99(m，1H)，3.22-3.09(m，4H)，1.26-1.18(m，18H).

MS(ESI m/z)：612(M+H)

RT(min)：1.31

Comparative example 3

The following compounds were obtained according to the methods described in Bioorganic & Medicinal Chemistry, volume 17, No. 17, pages 6340 to 6347, and 2009.

[ chemical formula 84]

S- (2- (((RS) - ((((2R, 3S, 4S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -3, 4-dihydroxytetrahydro-furan-2-yl) methoxy) (phenoxy) phosphoryl) oxy) ethyl) 2, 2-dimethylthiopropionate

¹H-NMR(CD₃OD)：7.82-7.68(m，1H)，7.45-7.32(m，2H)，7.28-7.18(m，3H)，6.27-6.19(m，1H)，5.84-5.73(m，1H)，4.58-4.30(m，2H)，4.29-4.08(m，4H)，4.06-3.99(m，1H)，3.23-3.12(m，2H)，1.23-1.18(m，9H).

MS(ESI m/z)：544(M+H)

RT(min)：1.16

Comparative example 4

[ chemical formula 85]

First step of

The following compounds were obtained in the same manner as in reference example 1.

Tert-butyl tert-butoxycarbonyl (1- ((2R, 4S, 5R) -4- ((tert-butoxycarbonyl) oxy) -3, 3-difluoro-5- (hydroxymethyl) -tetrahydrofuran-2-yl) -2-oxo-1, 2-dihydropyrimidin-4-yl) carbamate

MS(ESI m/z)：564(M+H)

RT(min)：1.72

Second step of

The following compounds were obtained in the same manner as in the second step of example 11.

((2R, 3S, 5R) -5- (4- (bis (tert-butoxycarbonyl) amino) -2-oxopyrimidin-1 (2H) -yl) -3- ((tert-butoxycarbonyl) oxy) -4, 4-difluorotetrahydrofuran-2-yl) methyl dihydrogenphosphate

MS(ESI m/z)：644(M+H)

RT(min)：1.35

Third Process

The following compounds were obtained in the same manner as in the first step of example 11.

((2R, 3S, 5R) -5- (4-amino-2-oxopyrimidin-1 (2H) -yl) -4, 4-difluoro-3-hydroxytetrahydrofuran-2-yl) methyl bis (pivaloyloxymethyl) phosphate

¹H-NMR(CD₃OD)：7.61(d，1H，J＝7.9Hz)，6.31-6.21(m，1H)，5.95(d，1H，J＝7.9Hz)，5.73-5.63(m，4H)，4.53-4.32(m，1H)，4.30-4.14(m，1H)，4.13-4.03(m，1H)，1.26-1.18(m，18H).

MS(ESI m/z)：572(M+H)

RT(min)：1.20

Test examples Gemcitabine-resistant cell proliferation inhibition test

Gemcitabine-resistant strains of pancreatic cancer cell line Capan-1(ATCC Number: HTB-79) were prepared by the method described in Neoplasia, Vol.12, pp.807-817, and 2010, and cell growth inhibition tests were performed using the compounds of the present invention and the compounds of the comparative examples. The inhibition test of cell growth was carried out according to the following method.

In order to measure inhibition of proliferation by the compound, the number of whole cells was quantified based on the whole-cell ATP concentration using CellTiterGlo (Promega) reagent using firefly luciferase. Capan-1 was suspended in IMDM medium (Life technologies) containing penicillin/streptomycin (pen/strep) and 20% FBS, adjusted to 33333/mL, and seeded at 90. mu.L (3000) per 1 well in 96-well plates (Comming).

The cells were grown under standard cell growth conditions (37 ℃ C., 5% CO)₂) The whole culture was incubated for 24 hours, and after adding 10. mu.L of a gradient dilution of the compound or 0.1% DMSO (solvent control), the cells were cultured under standard cell proliferation conditions (37 ℃, 5% CO)₂) Then, the culture was propagated for 72 hours. To determine whole cell proliferation, luminescence counts (relative light units, RLU) were quantified after addition of equal volumes of CellTiter Glo reaction solution to each well according to the instructions for CellTiter Glo. IC relating to inhibition of proliferation₅₀The RLU signal exhibited by the DMSO solvent control after 72 hours of incubation was defined as 0% inhibition, corresponding to the concentration of compound in solution that gave 50% inhibition of whole cell proliferation for the DMSO solvent control. Each data point was obtained using two replicate samples.

IC₅₀Calculated by the following method.

The results of the test substance treatment groups were plotted with the logarithm of the cell treatment concentration (nmol/L) as the X-axis and the cell proliferation inhibition (%) as the Y-axis. The straight line connecting the nearest 2 points of the clamped cell growth inhibition rate Y of 50 (%) was obtained, and the 50% cell growth Inhibition Concentration (IC) defined by the straight line was calculated₅₀Value). For the results of the control substance group, IC was also determined by the same method₅₀The value is obtained.

IC₅₀The results of the total of 3 tests were calculated, and the average and standard deviation of the 3 tests were determined. The results are shown in the following table.

Evaluation criteria

[ Table 23]

The compounds of the present invention have excellent cell proliferation inhibitory activity against gemcitabine-resistant strains of tumor cells.

Industrial applicability

The thionucleoside derivative or salt thereof of the present invention has an excellent proliferation inhibitory activity against tumor cells, and is useful as a therapeutic agent for tumors. Furthermore, the thionucleoside derivative or a salt thereof of the present invention has an excellent growth inhibitory activity against gemcitabine-resistant tumor cells, and is useful as a therapeutic agent for gemcitabine-resistant tumors.

Claims

1. A thionucleoside derivative represented by the general formula [1] or a salt thereof:

in the formula (I), the compound is shown in the specification,

R¹represents an optionally protected hydroxyl group, an optionally substituted C_1-20Alkoxy, optionally substituted C_3-8Cycloalkoxy group, aryloxy group which may be substituted, heterocyclic oxy group which may be substituted, or amino group which may be substituted；

R²Represents optionally substituted C_1-20Alkoxy, optionally substituted C_3-8A cycloalkoxy group, an aryloxy group which may be substituted, a heterocyclic oxy group which may be substituted, or an amino group which may be substituted;

or R¹And R²Or may form a 5-to 10-membered nitrogen-containing/phosphorus-containing heterocycle which may be substituted, a 5-to 10-membered oxygen-containing/phosphorus-containing heterocycle which may be substituted, or a 5-to 10-membered nitrogen-containing/oxygen-phosphorus-containing heterocycle together with the phosphorus atom to which they are bonded;

R³represents a hydrogen atom;

or R²And R³Or with R²Bonded phosphorus atom, oxygen atom, methylene group, 2 carbon atoms constituting tetrahydrothiophene ring and R³The bonded oxygen atoms form together a 6-to 10-membered oxygen-containing phosphorus heterocycle which may be substituted.

2. The thionucleoside derivative or a salt thereof according to claim 1,

R¹a hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6Alkoxy, aryloxy which may be substituted with 1 or more substituents selected from substituent group a, heterocyclic oxy which may be substituted with 1 or more substituents selected from substituent group a, or amino which may be substituted with 1 or more substituents selected from substituent group a,

substituent group a:

a halogen atom; a hydroxyl group which may be protected; a cyano group; a nitro group; a carbamoyl group; an oxo group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkyl group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkyl disulfide group; c which may be substituted by 1 or more substituents selected from substituent group B_2-6An alkenyl group; c which may be substituted by 1 or more substituents selected from substituent group B_2-6An alkynyl group; c which may be substituted by 1 or more substituents selected from substituent group B_3-8A cycloalkyl group; can be selectedC substituted from 1 or more substituents of substituent group B_3-8Cycloalkyl disulfide group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkoxy group; c which may be substituted by 1 or more substituents selected from substituent group B_1-6An alkoxycarbonyloxy group; c which may be substituted by 1 or more substituents selected from substituent group B_3-8A cycloalkoxycarbonyl group; aryl which may be substituted with 1 or more substituents selected from substituent group B; an aryl disulfide group which may be substituted with 1 or more substituents selected from substituent group B; a heterocyclic group which may be substituted with 1 or more substituents selected from substituent group B; a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group B; an acyloxy group which may be substituted by 1 or more substituents selected from the substituent group B; an acylthio group which may be substituted with 1 or more substituents selected from substituent group B; aminocarbonyloxy which may be substituted with 1 or more substituents selected from substituent group B; or an aminocarbonylthio group which may be substituted with 1 or more substituents selected from substituent group B,

substituent group B:

a halogen atom; a hydroxyl group; a cyano group; a nitro group; a carboxyl group; a carbamoyl group; a hydroxymethyl group; c_1-6An alkyl group; c_2-6An alkenyl group; c_2-6An alkynyl group; c_3-8A cycloalkyl group; c_1-6An alkoxy group; c_1-6An alkoxycarbonyl group; c_3-8A cycloalkoxycarbonyl group; aromatic C_1-6An alkoxycarbonyl group; an aryl group; an aryloxy group; heterocyclic oxy which may be substituted with 1 or more substituents selected from hydroxy and hydroxymethyl; aromatic C_1-6An alkoxy group; or an acyloxy group.

3. The thionucleoside derivative or a salt thereof according to claim 1 or 2, wherein,

R²is C which may be substituted by 1 or more substituents selected from substituent group A_1-6Alkoxy, aryloxy which may be substituted with 1 or more substituents selected from substituent group A, or ammonia which may be substituted with 1 or more substituents selected from substituent group AAnd (4) a base.

4. The thionucleoside derivative or salt thereof according to any one of claims 1 to 3,

R¹a hydroxyl group which may be protected, C which may be substituted with 1 or more substituents selected from the substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, a heterocyclic oxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a; r²Is C which may be substituted by 1 or more substituents selected from substituent group A_1-6An alkoxy group, an aryloxy group which may be substituted with 1 or more substituents selected from substituent group a, or an amino group which may be substituted with 1 or more substituents selected from substituent group a.

5. The thionucleoside derivative or salt thereof according to any one of claims 1 to 4,

R¹is aryloxy which may be substituted by 1 or more substituents selected from substituent group A, R²Is an amino group which may be substituted with 1 or more substituents selected from substituent group a, or,

6. The thionucleoside derivative or a salt thereof according to claim 1, wherein the thionucleoside derivative is selected from the group consisting of,

(2S) -isopropyl 2- (((2RS, 4aR, 6R, 7S, 7aS) -6- (4-amino-2-oxopyrimidin-1 (2H) -yl) -7-fluoro-2-oxide tetrahydro-4H-thieno [3, 2-d ] [1, 3, 2] dioxaphosphino-2-yl) amino) propionate.

7. A pharmaceutical composition comprising the thionucleoside derivative or a salt thereof according to any one of claims 1 to 6.

8. The pharmaceutical composition of claim 7, for use in the treatment of a tumor.

9. The pharmaceutical composition according to claim 7 for use in the treatment of lung cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, pancreatic cancer, breast cancer, renal cancer, bladder cancer, uterine cancer, osteosarcoma, melanoma, leukemia, multiple myeloma, or malignant lymphoma.