KR20040036417A - Thionucleoside derivatives having potent anti-cancer activity and the pharmaceutical compositions containing the same - Google Patents

Abstract

The present invention relates to a thionucleoside derivative compound having a novel chemical structure capable of mediating cancer and inflammatory diseases, and a pharmaceutical composition containing the compound, comprising a ligand for selectively activating adenosine A ₃ receptor. Designed and pharmacologically validated, it provides a pharmaceutical composition useful for the prevention and treatment of cancer and inflammatory diseases.

Description

Thionucleoside derivatives having potent anti-cancer activity and the pharmaceutical compositions containing the same

It is an object of the present invention to provide useful new chemical structures of thionucleoside derivative compounds having anticancer activity.

Adenosine is a substance that performs many physiological functions through receptors on specific cell membranes. Extracellular adenosine acts as a neurotransmitter in many physiological systems, and generally compensates for the hyperactivity of a given organ. Protects against the harmful effects of stress (Jacobson, KA et al . ; J. Med. Chem. , 35 , pp407-422, 1992). This is a partially generated negative feedback loop that attempts to reduce the energy demand of the cells by adenosine produced by the breakdown of intracellular or extracellular adenosine triphosphate (ATP) and increase the supply of oxygen. Adenosine is important for maintaining the homeostasis of essential organs such as the brain, heart and kidneys.For example, the administration of adenosine agonists from the outside to the brain has proved to be neuroprotective, and can be pain, cognition, exercise or sleep. It is also known to be involved.

Adenosine receptors are currently classified as P1 and P2 receptors through pharmacological studies and molecular cloning, respectively. Adenosine acts as a substrate for P1 receptors, and ATP, ADP, UTP, and UDP act as substrates for physiological activity. Among them, four different subtypes of adenosine receptors were identified. according to affinity, the body distribution of the ligand (ligand), acting like path is classified as a _1, a ₂ or a _3, a ₂ is divided back to the a _2a and a _2b. These adenosine receptors are a class of G-protein-coupled receptors. Adenosine A ₁ , A _2a and A _2b receptors have been identified pharmacologically using many selective ligands, but adenosine A _{The three} receptors were recently identified in 1992 (Zhou, Q. Y, et al . ; Proc. Natl. Acad. Sci. USA , 89 , pp7432-7436, 1992), confirming the pathophysiological function of these receptors. There is a lot of research to do.

Adenosine A ₁ and A ₂ receptor agonists are mainly derivatives of adenosine and have been studied as antihypertensive agents, antipsychotics, arrhythmia drugs, lipoprotein metabolism inhibitors (diabetic agents), brain protectants, and antagonists are xanthine derivatives Or a combination of several disease rings have been developed as an asthma treatment, antidepressant, arrhythmia treatment, kidney protector, Parkinson's disease treatment, intelligence development. Nevertheless, it is currently commercialized for adenosine itself, which is used for the treatment of supraventricular tachycardia, and dipyridamole, an adenosine transport inhibitor used as an adjuvant of warfarin to prevent blood coagulation following cardiac surgery. dipyridamole). This development was not successful because the adenosine receptors are spread all over the body because of the various pharmacological actions involved when the receptors are activated, and there is no compound that can activate only adenosine receptors of the desired tissue.

Adenosine A ₃ receptor is _{one of the} adenosine receptors A ₁ and A ₂ , which is widely known as a recently known receptor, unlike the adenosine receptors A ₁ and A _2, and many studies have been conducted to develop selective ligands. To pharmacologically study the adenosine A ₃ receptor, [ ¹²⁵ I] ABA (N ^6- (4-amino-3- ¹²⁵ I [iodo] benzyl) adenosine), [ ¹²⁵ I] APNEA ([ ¹²⁵ I] N ⁶ Three radiolabeled ligands are used, either 2- (4-aminophenyl) ethyladenosine) or [ ¹²⁵ I] AB-MECA ([ ¹²⁵ I] 4-aminobenzyl-5'-N-methylcarboxyamidoadenosine). When expressed in Chinese Hamster Ovary (CHO) cells, the A ₃ receptor is an inhibitor of adenylyl cyclase, an enzyme that produces cAMP from ATP, and the A ₃ receptor is activated by agonists. When activated, it was demonstrated that the brain activates GTP-dependent phospholipase C, an enzyme that breaks down phosphatidyl inositol to produce inositol phosphate and DAG. Ramkumar, V., et al . ; J. Biol. Chem. , 268 , pp168871-168890, 1993: Abbracchio, MP et al . ; Mol. Pharmacol. , 48 , pp1038-1045, 1995). These findings explain the possibility of response pathways by A ₃ activation in brain ischemia, because this secondary transporter system implies the pathway of neuronal injury in cerebral ischemia. In addition, adenosine A ₃ agonists are known to have a protective effect against brain diseases such as epilepsy and a protective effect against the heart. Activation of the adenosine A ₃ receptor causes the release of histamine-inducing agents from mast cells and constricts the bronchus, and high concentrations of agonists or antagonists can cause apoptosis in immune cells. . Agonists of the A ₃ receptor inhibit the release of tumor necrosis factor (TNF-α), which is an inflammation mediator, and also MIP-1α, interleukin-12, and interferon-γ. Suppresses the creation of). Therefore, adenosine A ₃ antagonists have potential to be developed as anti-inflammatory and asthma treatments, and if a compound with pharmacological selectivity can be developed, it will be possible to develop new therapeutic drugs for various diseases such as asthma, inflammation, cerebral ischemia, heart disease and cancer. .

Among the materials studied so far, the selective adenosine A ₃ ligand is N ^6- (3-iodobenzyl) -5 '-( N -methylcarbamoyl) adenosine ( N ^6- (3- iodobenzyl) -5 '-( N -methylcarbamoyl) adenosine; IB-MECA) and N ^6- (3-iodobenzyl) -2-chloro-5'-( N -methylcarbamoyl) adenosine ( N ) ^6- (3-iodobenzyl) -2-chloro-5 '-( N -methylcarbamoyl) adenosine; CI-IB-MECA) is a representative substance showing high selectivity for the A ₃ receptor compared to the adenosine A ₁ and A ₂ receptors. Can be.

In order to show affinity for the A ₃ receptor through analysis of various previous studies, an N -methylcarbamoyl group must be present at the 5th position of the sugar, and the base portion of the 6th position of the purine is an arylamino group or an alkyl group. It must be substituted with an amino group. Until now, no studies have been conducted to compare affinity by introducing substituents other than hydroxy groups at positions 2 or 3 of ribose. Thus, adenosine A ₃ receptors have been synthesized by synthesizing substances substituted with various substituents at positions 2 or 3 of the sugar. Correlations were studied by measuring and comparing the affinity for.

As such, pharmacological studies are performed by taking ligands derived from structure-activity relationship studies, and measuring affinity for adenosine A ₃ receptors and selectivity for adenosine A ₁ or A ₂ receptors to selectively select adenosine A ₃ receptors. By developing a ligand that acts, there is no side effect and the pharmacological effect is increased, and the present invention was completed by completing a thionucleoside derivative compound effective for anti-inflammatory and anticancer agents.

The present invention seeks to provide useful novel chemical structures of thionucleoside derivative compounds that can mediate cancer and inflammatory diseases.

In order to achieve the above object, the present invention provides a compound represented by the following general formula (I), a pharmaceutically acceptable salt thereof, and an isomer thereof as a thionucleoside derivative.

(Ⅰ)

In the above formula,

X is a sulfur atom or an oxygen atom;

R ₁ is a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, a benzyl group, habenzyl, or a phenylalkyl group;

R ₂ is a hydrogen atom, a halogen group, an alkoxy group, an alkenyl group, an alkynyl group, an alkylthio group or a thi group;

R ₃ and R ₃ ′ are hydroxyalkyl groups, alkoxycarbonyl groups or alkylaminocarbonyl groups of 1 to 5 carbon atoms, provided that R ₃ and R ₃ ′ do not have the same substituent at the same time;

R ₄ is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms.

The present invention also includes a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient.

In formula (I), R ₁ is a 3-iodobenzyl group, R ₂ is a chlorine atom, R ₃ is a methylaminocarbonyl group, R ₃ ′ and R ₄ are hydrogen atoms, and X is a sulfur atom As the selected compound, a compound represented by the following general formula (II) and an isomer thereof are included, and preferably (2R, 3S, 4R) -2- [2-chloro-6- (3-iodo-benzylamino) -Purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide.

(Ⅱ)

In the general formula (I), R ₁ is a hydrogen atom, a methyl group or 3-iodobenzyl group, R ₂ is a chlorine atom, R ₃ and R ₄ are hydrogen atoms, R ₃ 'is a methylaminocarbonyl group or X is a hydroxymethyl group, X is a compound selected from sulfur atoms, and includes a compound represented by the following general formula (III) and an isomer thereof, preferably

(2R, 3R, 4S, 5R) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4 -Dior,

(2R, 3R, 4S, 5R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol,

(2R, 3R, 4S, 5R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol,

(2S, 3S, 4R, 5R) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide,

(2S, 3S, 4R, 5R) -5- (2-Chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide,

(2S, 3S, 4R, 5R) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2 -Carboxylic acid methylamide.

(Ⅲ)

In the general formula (I), R ₁ is a 3-iodobenzyl group, R ₂ is a chlorine atom, R ₃ ′ is a hydroxymethyl group, R ₃ and R ₄ are hydrogen atoms, X is selected from sulfur atoms As the compound, a compound represented by the following formula (IV) and an isomer thereof are included, and preferably (2S, 3R, 4S, 5R) -2- [2-chloro-6- (3-iodo-benzylamino) -Purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol.

(Ⅳ)

Another object of the present invention is to provide a method for preparing the compound of the general formula (I), which may be chemically synthesized by the method shown in Schemes 1 to 7, but is not limited thereto. The following schemes represent the preparation steps of representative compounds of the present invention, and other compounds may be prepared by appropriate changes in reagents and starting materials known to those skilled in the art.

As in Scheme 1, the synthesis of the compound of Formula 4 can be obtained by reaction in the presence of anhydrous copper (II) sulfate using anhydrous acetone as a reagent and a solvent with concentrated sulfuric acid from D-gulonic γ-lactone as an acid catalyst. In addition to the concentrated sulfuric acid, the reaction may use inorganic acids such as hydrochloric acid gas or organic acids such as p -toluenesulfonic acid, and dehydrating agents such as molecular sieve or anhydrous magnesium sulfate in addition to copper (II) sulfate as a dehydrating agent. It is also possible. Compound 5 may be reacted with lithium compound 4 to synthesize compound 5, and it is preferable to use an inert solvent such as ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, and sodium instead of lithium aluminum hydride. It is also possible to use metal hydrides such as borohydride. The obtained compound 5 may be reacted with methanesulfonyl chloride to obtain compound 6, and an amine reagent such as pyridine and triethylamine (TEA) may be used as a solvent and a reagent, or ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran , N, N-dimethylformamide and it is preferable to use a mixture with an inert solvent such as, N, N as a reaction promoter - it is also possible to use dimethylaminopyridine or 2,6-lutidine. The compound of Formula 7 may be obtained by reacting compound 6 with sodium sulfide, and it is also possible to use sodium alkoxide after substitution reaction with thio ester such as metal thioacetate instead of sodium sulfide. As the solvent, solvents such as N, N -dimethylformamide, dimethyl sulfoxide and the like are preferable, and it is also possible to react with lower alcohols such as methanol and ethanol, and water or an inert organic solvent can be mixed and used therein. Compound 8 can be obtained by reacting the prepared compound 7 with an aqueous acetic acid solution.Instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid, an organic acid such as p -toluenesulfonic acid, or water or a lower alcohol such as methanol can be used alone or with an organic solvent. It is also possible to mix and use it as a solvent. The compound of Formula 9 may be obtained by reacting Compound 8 with red tetraacetate, and it is also possible to use sodium metaperiodate at a low temperature instead of red tetraacetate, ethyl acetate, ethyl ether, petroleum ether, dichloro Preference is given to using inert solvents such as methane, tetrahydrofuran and N, N -dimethylformamide.

As in Scheme 2, the compound of Formula 10 may be obtained by reacting Compound 9 of Scheme 1 and pyridinium dichromate in a N , N -dimethylformamide solvent, and the compound of Formula 11 Obtained by reacting dimethyl sulfate in the presence of potassium carbonate, methyl halides or diazomethane, such as methyl iodine, in place of dimethyl sulfate, and inorganic bases such as sodium carbonate or DBU or n -butyllithium in place of potassium carbonate The same organic base may be used, and acetone may be used as a solvent, or an organic solvent such as tetrahydrofuran or dioxane may be used.

The compound of formula 12 is obtained by reacting a compound of formula 11 with a methylamine tetrahydrofuran solution or aqueous solution, and the solvent is tetrahydrofuran single solvent or ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N , N -dimethyl It is also possible to use an inert solvent such as formamide in admixture with water, and the compound of formula 13 is obtained by reacting the compound of formula 12 with m -chloroperbenzoic acid, instead of m -chloroperbenzoic acid, sodium metaperiodate, It is also possible to use t -butyl peroxide, peracetic acid, hydrogen peroxide and the like, and inert solvents such as ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N , N -dimethylformamide alone or inert It is preferable to use a mixed solvent of a solvent and water.

The compound of formula 14 is obtained by reacting the compound of formula 13 with silylated 2,6-dichloropurine in the presence of trimethylsilyl trifluoromethanesulfonate, and the solvent is dichloroethane, chloroform, acetonitrile, dichloromethane The same inert solvent is preferred, and the silylated 2,6-dichloropurine can be reacted with 2,6-dichloropurine and hexamethyldisilazane as an reagent and solvent under an ammonium sulfate catalyst or directly with 2,6-dichloropurine. It can be obtained by reaction with N , O -bis (trimethylsilyl) acetamide. The compound of formula 15 is obtained by reacting compound of formula 14 with 3-iodobenzylamine hydrochloride in the presence of triethylamine base, and instead of triethylamine pyridine, N , N -dimethylaminopyridine or 2,6-ruti It is also possible to use organic bases such as dean, and preferred solvents include lower alcohols such as methanol and ethanol or solvents such as 1,4-dioxane, tetrahydrofuran and chloroform. The compound of formula (16) is obtained by reacting the compound of formula (15) with an aqueous acetic acid solution. Instead of acetic acid, inorganic acids such as sulfuric acid or hydrochloric acid, organic acids such as p -toluenesulfonic acid, water or lower alcohols such as methanol alone or organic It is also possible to mix with a solvent and use it as a solvent.

As in Scheme 3, the compound of Formula 17 can be obtained by reacting the compound of Formula 9 with sodium borohydride, it is preferable to use a lower alcohol, such as methanol, ethanol, these lower alcohol and dichloromethane, It is also possible to make it react in mixed solvent with solvents, such as tetrahydrofuran and chloroform. The compound of formula 18 may be obtained by reacting the compound of formula 17 with benzoyl chloride, and using an amine reagent such as pyridine, triethyl amine as a solvent and reagent, or ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N It is preferable to use it in admixture with an inert solvent such as N -dimethylformamide or chloroform. It is also possible to use N , N -dimethylaminopyridine or 2,6-lutidine as the reaction promoter.

The compound of formula 19 may be obtained by reacting the compound of formula 18 with m -chloroperbenzoic acid, using sodium metaperiodate, t -butylperoxide, peracetic acid, hydrogen peroxide and the like instead of m -chloroperbenzoic acid. It is possible to use an inert solvent such as ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N , N -dimethylformamide alone or a mixed solvent of these inert solvents and water as the solvent. The compound is obtained by reacting the compound of formula 19 with acetic anhydride, preferably using acetic anhydride as a solvent and reagent. It is also preferable to increase the reactivity by adding acetate ions such as sodium acetate or tetrabutylammonium acetate.

Compounds of formulas (21) and (22) can be obtained by reacting a compound of formula (20) with a silylated 2,6-dichloropurine in the presence of trimethylsilyl trifluoromethanesulfonate, and as solvents, dichloroethane, chloroform, acetonite Inert solvents such as reyl and dichloromethane are preferred, and the silylated 2,6-dichloropurine is reacted with 2,6-dichloropurine and hexamethyldisilazane as an reagent and solvent under an ammonium sulfate catalyst or 2,6 It can be obtained by directly reacting dichloropurine with N , O -bis (trimethylsilyl) acetamide, and using an inorganic acid such as tin (IV) chloride as a catalyst instead of trimethylsilyl trifluoromethanesulfonate as a catalyst. It is possible.

The compound of formula 23 may be obtained by reacting compound of formula 21 with 3-iodobenzylamine hydrochloride in the presence of triethylamine base, and instead of triethylamine pyridine, N , N -dimethylaminopyridine or 2,6 It is also possible to use an organic base such as lutidine, and as a solvent, a lower alcohol such as methanol or ethanol or a solvent such as 1,4-dioxane, tetrahydrofuran or chloroform is preferable, and the compound of formula 24 is Obtained by reacting the compound of 22 with 3-iodobenzylamine hydrochloride in the presence of a triethylamine base, and instead of triethylamine an organic such as pyridine, N , N -dimethylaminopyridine or 2,6-lutidine It is also possible to use a base, and the solvent may be a lower alcohol such as methanol or ethanol or a solvent such as 1,4-dioxane, tetrahydrofuran or chloroform. It is.

The compound of formula 25 is reacted with a compound of formula 23 and a metal alkoxide such as sodium methoxide and sodium ethoxide in a lower alcohol solvent such as methanol or ethanol or as a mixed solvent with an inert solvent such as dichloromethane or chloroform. Or a compound obtained by reacting ammonia with an alcohol solvent such as methanol or ethanol, or an inorganic base such as sodium carbonate or sodium bicarbonate in a mixed solvent of a compound of formula 23 with water and a lower alcohol. The material can be obtained by reacting with an aqueous acetic acid solution, and instead of acetic acid, inorganic acids such as sulfuric acid or hydrochloric acid, or organic acids such as p -toluenesulfonic acid can be used alone or as a solvent by mixing a lower alcohol such as methanol with an organic solvent. It is also possible.

The compound of formula 26 is reacted with a compound of formula 24 and a metal alkoxide such as sodium methoxide and sodium ethoxide in a lower alcohol solvent such as methanol or ethanol or as a mixed solvent with an inert solvent such as dichloromethane or chloroform. Or a compound obtained by reacting a compound of formula 24 with ammonia in an alcohol solvent such as methanol or ethanol, or an inorganic basic substance such as sodium carbonate or sodium bicarbonate in a mixed solvent of a compound of formula 24 with water and a lower alcohol. The material can be obtained by reacting with an aqueous acetic acid solution, and instead of acetic acid, inorganic acids such as sulfuric acid or hydrochloric acid, organic acids such as p -toluenesulfonic acid, water or lower alcohols such as methanol can be used alone or as a solvent by mixing with an organic solvent. It is also possible to use.

As in Scheme 4, using the compound of formula 21 as a starting material, a compound of formula (III) wherein R ₁ is a hydroxymethyl group, R ₂ is a methylamino group, and R ₃ is a chloro group and R ₁ is a hydroxide A method for preparing a compound of thionucleoside in which a oxymethyl group, R ₂ is an amino group, and R ₃ is a chloro group is shown in Scheme 4 below.

The compound of formula 27 may be obtained by reacting the compound of formula 21 with methylamine tetrahydrofuran solution or with methylamine aqueous solution, preferably in a closed container of metal or special material, and the compound of formula 28 By reacting the compound of formula 27 with an aqueous acetic acid solution, or instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid, or an organic acid such as p -toluenesulfonic acid, alone or in combination with an organic solvent, a lower alcohol such as methanol, The resulting material is reacted with a metal alkoxide such as sodium methoxide and sodium ethoxide in a lower alcohol solvent such as methanol or ethanol or as a mixed solvent with an inert solvent such as dichloromethane or chloroform, or ammonia can be reacted with methanol or ethanol. In an alcoholic solvent such as water or low It can be obtained by reacting with inorganic basic substances such as sodium carbonate and sodium bicarbonate in a mixed solvent with a tertiary alcohol.

The compound of formula 29 may be obtained by reacting the compound of formula 21 with a lower alcohol solution such as saturated ammonia ethanol solution or methanol solution, and also reacting with saturated ammonia 1,4-dioxane solution. Is a solvent obtained by reacting a compound of Formula 29 with an aqueous acetic acid solution, or instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid, an organic acid such as p -toluenesulfonic acid, or a lower alcohol such as methanol alone or in combination with an organic solvent. The resulting material is reacted with a metal alkoxide such as sodium methoxide and sodium ethoxide in a lower alcohol solvent such as methanol, ethanol or as a mixed solvent with an inert solvent such as dichloromethane, chloroform or ammonia in methanol. In an alcohol solvent such as ethanol or water and Class in the mixed solvent of the alcohol may be obtained by reacting with an inorganic basic substance such as sodium carbonate, sodium bicarbonate.

As in Scheme 5, the compound of formula 31 is reacted with the compound of formula 29 and an aqueous acetic acid solution, or instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid, or an organic acid such as p -toluenesulfonic acid is lower than water or methanol. It can be obtained by reacting the alcohol alone or in a solvent mixed with an organic solvent, the compound of formula 32 can be obtained by reacting the compound of formula 31 with tert - butylchlorodimethylsilane in the presence of imidazole base, and imidazole Instead, it is also possible to use organic bases such as triethylamine, pyridine, N , N -dimethylaminopyridine or 2,6-lutidine, and as solvents N , N -dimethylaminoformamide, dichloromethane, tetrahydro Preference is given to using inert solvents such as furan and chloroform.

The compound of formula 33 is reacted with a compound of formula 32 and a metal alkoxide such as sodium methoxide, sodium ethoxide in a lower alcohol solvent such as methanol, ethanol or as a mixed solvent with an inert solvent such as dichloromethane, chloroform, or ammonia Is reacted with an alcohol solvent such as methanol or ethanol, or by reacting with an inorganic basic substance such as sodium carbonate or sodium bicarbonate in a mixed solvent of water and a lower alcohol. The resultant obtained by reacting pyridinium dichromate in a N , N -dimethylformamide solvent can be obtained by reacting in the presence of dimethyl sulfate and potassium carbonate, and instead of dimethyl sulfate, methyl halide or diazomethane such as methyl iodine; Instead of potassium carbonate, it is also possible to use an inorganic base such as sodium carbonate or an organic base such as dibutyl oil (DBU) or n -butyllithium, and use acetone as a solvent or use an organic solvent such as tetrahydrofuran or dioxane. It is also possible to use.

The compound of formula 35 may be obtained by reacting a compound of formula 34 with a methylamine tetrahydrofuran solution or an aqueous solution. The solvent may be tetrahydrofuran single solvent or ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N , N It is also possible to use an inert solvent such as dimethylformamide in admixture with water, and the compound of formula 36 can be obtained by reacting the compound of formula 35 with tetrabutylammonium fluoride, triethylammonium instead of tetrabutylammonium fluoride It is also preferable to use a reagent capable of providing fluoride such as mamin trihydrofluoride, hydrogen fluoride-pyridine, or an organic acid such as p -toluenesulfonic acid or an inorganic acid such as hydrochloric acid.

As in Scheme 6, the compound of Formula 37 is reacted with the compound of Formula 27 and an aqueous acetic acid solution, or instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid or an organic acid such as p -toluenesulfonic acid is substituted with water or a lower alcohol such as methanol. It can be obtained by reacting alone or in a solvent mixed with an organic solvent, the compound of formula 38 can be obtained by reacting the compound of formula 37 with tert - butylchlorodimethylsilane in the presence of imidazole base, instead of imidazole It is also possible to use organic bases such as triethylamine, pyridine, N , N -dimethylaminopyridine or 2,6-lutidine, and the solvents include N , N -dimethylaminoformamide, dichloromethane, tetrahydrofuran, Preference is given to using inert solvents such as chloroform.

The compound of formula 39 is reacted with a compound of formula 38 and a metal alkoxide such as sodium methoxide, sodium ethoxide in a lower alcohol solvent such as methanol, ethanol or as a mixed solvent with an inert solvent such as dichloromethane, chloroform, or ammonia Is reacted with an alcohol solvent such as methanol or ethanol, or by reacting with an inorganic basic substance such as sodium carbonate or sodium bicarbonate in a mixed solvent of water and a lower alcohol. The resultant obtained by reacting pyridinium dichromate in a N , N -dimethylformamide solvent can be obtained by reacting in the presence of dimethyl sulfate and potassium carbonate, and instead of dimethyl sulfate, methyl halide or diazomethane such as methyl iodine; Instead of potassium carbonate, it is also possible to use inorganic bases such as sodium carbonate or organic bases such as dibutyl (DBU) or n -butyllithium, and use acetone as a solvent or organic solvents such as tetrahydrofuran and dioxane. It is also possible.

The compound of formula 41 may be obtained by reacting a compound of formula 40 with a methylamine tetrahydrofuran solution or an aqueous solution. The solvent may be tetrahydrofuran single solvent or ethyl ether, petroleum ether, dichloromethane, tetrahydrofuran, N , N It is also possible to use an inert solvent such as dimethylformamide in admixture with water, and the compound of formula 42 may be obtained by reacting the compound of formula 41 with tetrabutylammonium fluoride, and triethyl instead of tetrabutylammonium fluoride It is also preferable to use a reagent capable of providing fluoride such as mamin trihydrofluoride and hydrogen fluoride-pyridine, or to use an organic acid such as p -toluenesulfonic acid or an inorganic acid such as hydrochloric acid.

As shown in Scheme 7, the compound of formula 43 reacts with the compound of formula 23 and an aqueous acetic acid solution, or instead of acetic acid, an inorganic acid such as sulfuric acid or hydrochloric acid or an organic acid such as p -toluenesulfonic acid is substituted with water or a lower alcohol such as methanol. It can be obtained by reacting alone or in a solvent mixed with an organic solvent, the compound of formula 44 can be obtained by reacting the compound of formula 43 with tert - butylchlorodimethylsilane in the presence of imidazole base, instead of imidazole It is also possible to use organic bases such as triethylamine, pyridine, N , N -dimethylaminopyridine or 2,6-lutidine, and the solvents include N , N -dimethylaminoformamide, dichloromethane, tetrahydrofuran, It is preferable to use an inert solvent such as chloroform, and the compound of formula 45 is a compound of formula 44 and sodium methoxa The metal alkoxide such as sodium ethoxide may be reacted in a lower alcohol solvent such as methanol or ethanol or as a mixed solvent with an inert solvent such as dichloromethane or chloroform, or the ammonia in an alcohol solvent such as methanol or ethanol. Or by reacting with inorganic basic substances such as sodium carbonate and sodium bicarbonate in a mixed solvent of water and lower alcohol.

The compound of formula 46 can be obtained by reacting a compound of formula 45 with a pyridinium dichromate in a N , N -dimethylformamide solvent in the presence of dimethyl sulfate and potassium carbonate, and methyl iodine instead of dimethyl sulfate. Methyl halides or diazomethanes, such as inorganic carbonates such as sodium carbonate or organic bases such as dibutyl (DBU) or n -butyllithium, may be used instead of potassium carbonate, and acetone may be used as a solvent, or tetra It is also possible to use an organic solvent such as hydrofuran and dioxane, and the compound of formula 47 may be obtained by reacting the compound of formula 46 with a methylamine tetrahydrofuran solution or an aqueous solution, and as a solvent, a tetrahydrofuran single solvent or Ethyl ether, petroleum ether, dichloromethane, tetrahydro It is also possible to use an inert solvent such as furan, N , N -dimethylformamide in admixture with water.

Compound 48 may be obtained by reacting compound 47 with tetrabutylammonium fluoride, providing fluoride such as triethylmamin trihydrofluoride, hydrogen fluoride-pyridine and the like instead of tetrabutylammonium fluoride It is also preferable to use a reagent which can be used, or to use an organic acid such as p -toluenesulfonic acid or an inorganic acid such as hydrochloric acid.

Since the compounds disclosed in the present invention have a sub-carbon, racemic mixtures of the general formula (I) and R or S-type stereoisomers are also included in the scope of the present invention, and the present invention includes compounds in the form of pharmaceutically acceptable salts. This includes them.

Pharmaceutically acceptable salts of the general formula (I) above include salts of acidic or basic groups which may be present in compounds of general formula (I), unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art It can be prepared through.

The compounds of formula (I) have asymmetric centers and therefore may exist in different enantiomeric forms, and all optical isomers and stereoisomers of compounds of formula (I) and mixtures thereof are also included within the scope of the present invention. Shall be. The present invention encompasses the use of racemates, one or more enantiomeric forms, one or more diastereomeric forms, or mixtures thereof, and includes methods or processes for the separation of isomers known in the art.

Another object of the present invention is a pharmaceutical having adenosine A ₃ receptor selective activity, which is contained as an active ingredient in an amount effective to alleviate cancer or inflammation, together with a compound of formula (I) and a pharmaceutically acceptable carrier, adjuvant or diluent. It is to provide a composition.

The cancer may include lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, anal muscle cancer, colon cancer, breast cancer, fallopian tube carcinoma, endometrial carcinoma and cervical cancer. Species, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine gland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis cancer, prostate cancer, chronic or acute leukemia, lymphocyte Lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, pituitary adenoma, or a combination of one or more of these cancers It means to include.

The above-mentioned inflammation is meant to include acute and chronic inflammatory diseases such as degenerative inflammation, exudative inflammation, purulent inflammation, hemorrhagic inflammation or proliferative inflammation.

The present invention also provides a composition for the prevention and treatment of cancer or inflammatory diseases comprising the above general formula (I) as an active ingredient and a pharmaceutically acceptable carrier.

For example, the compounds of the present invention can be dissolved in oil, propylene glycol or other solvents commonly used in the preparation of injectable solutions. Suitable carriers are not particularly limited but include, for example, saline, polyethylene glycol, ethanol, vegetable oils and isopropyl myristate, and the compounds of the present invention may be formulated as ointments or creams for topical application.

Hereinafter, the formulation method and the excipient will be described, but are not limited only to these examples.

Pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The compounds of the present invention can be prepared by dissolving, suspending or emulsifying the compound in a conventional saline solution, a water-soluble solvent such as 5% dextrose or a non-aqueous solvent such as vegetable oil, synthetic fatty acid glyceride, higher fatty acid ester or propylene glycol. It can be formulated as. Formulations of the present invention may include conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives.

Preferred dosages of the compounds of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, preferably, the compound of the present invention is administered at 0.001 to 100 mg / kg body weight per day, more preferably 0.01 to 30 mg / kg body weight. Administration may be administered once a day or may be divided several times. The compound of the present invention in the composition should be present in an amount of 0.0001 to 10% by weight, preferably 0.001 to 1% by weight relative to the total weight of the total composition.

The pharmaceutical compositions of the present invention can be administered to mammals such as mice, mice, livestock, humans, and the like by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerbroventricular injection.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited.

Example 1. (2 R , 3 S ,4 S , 5 R ) -2,3: 5,6-di- O -Isopropylidene-D-gulono-1,4-lactone compound preparation (1)

20.2 g (0.114 mol) of D-gulonic γ-lactone and 27 g (0.170 mol) of anhydrous copper (II) sulfate were suspended in 0.65 L of anhydrous acetone, and 1.7 ml of concentrated sulfuric acid was added at room temperature, followed by stirring for 24 hours. Filtering the calcium hydroxide to a pH of 7 focuses the resulting solid and the filtrate was concentrated under reduced pressure to 28.7g; (yield: 98%) (2 R, 3 S, 4 S, 5 R) -2,3: 5, 6-Di- O -isopropylidene-D-gulono-1,4-lactone was obtained in the oil phase, which was immediately used for the next reaction.

Example 2. {(4 R )-(2,2-dimethyl- [1,3] dioxolan-4-yl)}-{(4 R , 5 S )-(5-hydroxymethyl-2,2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -Methanol Compound Preparation (2)

In Example 1 (2 R, 3 S, 4 S, 5 R) -2,3: 5,6- di - O - isopropylidene-1,4-lactone no -D- den 25.1g (0.097mol ) Was dissolved in 450 ml of ethyl ether, and 7.2 g (0.190 mol) of lithium aluminum hydride was carefully added dropwise at 0 ° C. in several portions, and then the reaction mixture was stirred at room temperature for 10 hours, and continuously 7.2 ml of water at 0 ° C. Then, 7.2 ml of 15% aqueous sodium hydroxide solution, 22 ml of water, and 19 g of anhydrous magnesium sulfate were added in this order, and the filtrate was concentrated under reduced pressure, followed by elution (hexane: ethyl acetate = 1: 1), using silica gel. purification by column chromatography, on 24.1g; {(4 R) - ( 2,2- dimethyl- [1, 3] dioxolan-4-yl)} (yield 95%) {(4 R, 5 S )-(5-hydroxymethyl-2,2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -methanol was obtained in the oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.29 (d, 6H), 1.36 (s, 3H), 1.43 (s, 3H), 3.26 (s, 1H), 3.28 (s, 1H), 3.36-3.79 (m , 4H), 3.96-4.03 (m, 2H), 4.12-4.22 (m, 2H)

Example 3. {(4 R )-(2,2-dimethyl- [1,3] dioxolan-4-yl)}-{(4 S , 5 S )-(5-methanesulfonyloxymethyl-2,2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -Methyl ester compound preparation (3)

{(4 R) - (2,2- dimethyl- [1, 3] dioxolan-4-yl)} - in Example 2 {(4 R, 5 S ) - (5- hydroxymethyl-2, 4.1 g (0.016 mol) of 2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -methanol was dissolved in a mixed solvent of 1.0 l of dichloromethane and 25 ml of pyridine and adjusted to 0 ° C. 18 mL (0.236 mol) of sulfonyl chloride was added. After stirring for 5 hours at 0 ° C., saturated sodium bicarbonate water was added and the mixture was extracted with chloroform, and the extract was filtered under reduced pressure after drying. The concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 2: 1). purification by calligraphy 6.4g (yield 98%) of methanesulfonic acid {(4 R) - (2,2- dimethyl- [1, 3] dioxolan-4-yl)} - {(4 S, 5 S )-(5-methanesulfonyloxymethyl-2,2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -methyl ester was obtained in oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.37 (s, 3H), 1.39 (s, 3H), 1.46 (s, 3H), 1.52 (s, 3H), 3.10 (s, 3H), 3.18 (s, 3H ), 3.92-4.08 (m, 1H), 4.08-4.20 (m, 2H), 4.39-4.49 (m, 4H), 4.81-4.87 (m, 1H)

Example 4 (3a S ,4 R , 6a R ) -4-((4 R ) -2,2-dimethyl- [1,3] dioxolan-4-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxole compound preparations (4)

Methanesulfonic acid in Example 3 {(4 R) - ( 2,2- dimethyl- [1, 3] dioxolan-4-yl)} - {(4 S, 5 S) - (5- methanesulfonyl 10.1 g (0.024 mol) of oxymethyl-2,2-dimethyl- [1,3] dioxolan-4-yl)}-( S ) -methyl ester was dissolved in 260 ml of N , N -dimethylformamide, followed by sodium 8.9 g (0.036 mol) of sulfide was added and the reaction mixture was reacted at 100 ° C. for 3 hours. The reaction mixture was concentrated under reduced pressure, water was added to the concentrate, extracted with ethyl acetate, the organic layer was dried and filtered and the filtrate was concentrated under reduced pressure. The concentrate was purified by column chromatography on silica gel using eluent (hexane: ethyl acetate = 5: 1) to give 5.9 g (yield; 94%) of (3a S , 4 R , 6a R ) -4-(( 4 R ) -2,2-dimethyl- [1,3] dioxolan-4-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxole is in oil phase Was obtained.

¹ H-NMR (CDCl ₃ ) δ: 1.33 (d, 6H), 1.44 (s, 3H), 1.51 (s, 3H), 2.86 (d, 1H), 3.09 (m, 1H), 3.22 (s, 1H ), 3.76 (dd, 1H), 3.98 (m, 1H), 4.15 (dd, 1H), 4.93 (d, 2H)

Example 5 (1 R ) -1-((3a S ,4 R , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-yl) -ethane-1,2-diol compound (5)

(3a S , 4 R , 6a R ) -4-((4 R ) -2,2-dimethyl- [1,3] dioxolan-4-yl) -2,2-dimethyl-tetra of Example 4, above 150 ml of an aqueous 60% acetic acid solution was added to 5.4 g (0.020 mol) of hydro-thieno [3,4- d ] [1,3] dioxol, followed by stirring at room temperature for 50 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to obtain 3.4 g (yield; 75%) of ( 1R ) -1- ( (3a S, 4 R, 6a R) -2,2- dimethyl-tetrahydro-thieno [3,4- d] [1,3] dioxol-4-yl) -ethane-1,2-diol Obtained in oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.34 (s, 3H), 1.53 (s, 3H), 2.90 (dd, 1H), 3.09 (dd, 1H), 3.27 (dd, 1H), 3.58-3.65 (m , 2H), 3.69 (dd, 1H), 3.79 (dd, 1H), 4.15 (dd, 1H), 4.93 (m, 2H)

Example 6 (3a S ,4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carbaldehyde compound preparations (6)

(1 R) of Example 5 -1 - ((3a S, 4 R, 6a R) -2,2- dimethyl-tetrahydro-thieno [3,

4- d ] [1,3] dioxol-4-yl) -ethane-1,2-diol 2.5 g (11.2 mmol) was dissolved in ethyl acetate, and then red tetraacetate was added at 0 ° C, and the reaction mixture was 10 The mixture was stirred for 10 minutes, the reaction mixture was filtered, ethyl acetate was added to the filtrate, washed with saturated sodium bicarbonate water and dried. The filtrate was concentrated under reduced pressure and the eluate was concentrated (hexane: ethyl acetate = 5: 1). Purification by column chromatography on silica gel using hexane, 2.1 g (yield; 98%) of (3a S , 4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carbaldehyde was obtained in the oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.33 (s, 3H), 1.52 (s, 3H), 2.61 (dd, 1H), 2.87 (dd, 1H), 3.92 (s, 1H), 4.92 (t, 1H ), 5.10 (d, 1H), 9.43 (s, 1H)

Example 7. (3a S ,4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methyl ester compound synthesis (7)

(3a S , 4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,

4- d ] [1,3] dioxol-4-carbaldehyde 2.8 g (0.015 mol) was dissolved in 200 ml of anhydrous N , N -dimethylformamide, followed by the addition of 26.7 g (0.070 mol) of pyridinium dichromate. The mixed solution was stirred at room temperature for 20 hours. 0.3 liters of water was added, followed by extraction with 2.5 liters of ethyl acetate (0.5 liters x 5), followed by drying. The filtrate obtained by filtration was concentrated under reduced pressure to give a concentrate (3a S , 4 S , 6a R ) -2,2-dimethyl-. After obtaining tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid, the concentrate was dissolved in 65 ml of acetone, and 2 ml (21.14 mmol) of dimethyl sulfate and 1.2 g of potassium carbonate. (8.68 mmol) was added and stirred for 2 hours. Acetone was distilled off under reduced pressure, and the residue was dissolved in ethyl acetate, washed with water and brine, dried, filtered and the filtrate was concentrated under reduced pressure. The resulting mixture was evaporated (hexane: ethyl acetate = 3: 1). Purified by column chromatography on silica gel using 2.5 g (yield; 78%) of (3a S , 4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methyl ester was obtained in an oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.33 (s, 3H), 1.52 (s, 3H), 2.92 (d, 1H), 3.19 (dd, 1H), 3.72 (s, 3H), 3.80 (s, 1H ), 4.99 (m, 2 H)

Example 8 (3a S ,4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide compound synthesis (8)

1.1 g of (3a S , 4 S , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methyl ester of Example 7 (0.005 mol) was dissolved in 40 mL (0.080 mol) of 2N methylamine in tetrahydrofuran solution and stirred in a sealed container at 50 ° C. for 24 hours. The resulting concentrate after distillation under reduced pressure was purified by column chromatography on silica gel using eluent (hexane: ethyl acetate = 2: 1), and 953 mg (yield; 87%) of (3a S , 4 S , 6a R ). -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide was obtained in the oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.29 (s, 3H), 1.48 (s, 3H), 2.78 (d, 3H), 2.91 (d, 1H), 3.04 (dd, 1H), 3.72 (s, 1H ), 4.91 (t, 1H), 5.21 (d, 1H), 6.65 (br s, 1H)

Example 9 (3a S ,4 S , 5 R , 6a R )-And (3a S ,4 S , 5 S , 6a R ) -2,2-dimethyl-5-oxo-tetrahydro

-5λ ⁴ -Thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide compound synthesis (9)

(3a S, 4 S, 6a R) of Example 8 2,2-dimethyl-tetrahydro-thieno [3,4- d] [1,3] dioxol-4-carboxylic acid methylamide 120mg ( 0.55 mmol) was dissolved in 15 mL of dichloromethane and adjusted to −78 ° C., followed by the slow addition of a solution of 119 mg (0.55 mmol, 80% reagent) of m -chloroperbenzoic acid dissolved in 5 mL of dichloromethane. The reaction mixture was stirred at −78 ° C. for 45 minutes, and then saturated sodium bicarbonate water was added and extracted with dichloromethane. The organic layer was washed with saturated brine, dried, filtered and concentrated under reduced pressure. The resulting concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1.2), and 125 mg (yield; 97%). ) And (3a S , 4 S , 5 R , 6a R )-and (3a S , 4 S , 5 S , 6a R ) -2,2-dimethyl-5-oxo-tetrahydro-5λ ⁴ -thieno [ 3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide was obtained in the oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.23 (s, 3H), 1.37 (s, 3H), 2.80 (d, 3H), 3.21 (dd, 1H), 3.61 (d, 1H), 4.30 (s, 1H ), 5.06 (m, 2H), 7.12 (br s, 1H)

Example 10. (3a S ,4 R , 6a R ) -4- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide compound synthesis (10)

102 mg (0.54 mmol) of 2,6-dichloropurine and 20 mg (0.15 mmol) of ammonium sulfate were suspended in 10 ml of anhydrous hexamethyldisilazane and stirred under reflux for 6 hours. The reaction mixture was concentrated under reduced pressure to anhydrous state and concentrated. Was dissolved in 8 ml of dichloroethane and dissolved in 5 ml of dichloroethane (3a S , 4 S , 5 R , 6a R )-and (3a S , 4 S , 5 S , 6a R ) -2,2-dimethyl-5- Oxo-tetrahydro

110 mg (0.47 mmol) of -5λ ⁴ -thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide were added and adjusted to -10 ° C, followed by trimethylsilyl trifluoromethanesulfonate 0.10 mL (0.54 mmol) was added and the reaction mixture was stirred at −10 ° C. for 20 minutes and stirred at reflux for 4 hours. Saturated sodium bicarbonate was added, stirred for 15 minutes, extracted with dichloromethane, washed with saturated brine, dried, filtered and concentrated under reduced pressure. The eluate (hexane: ethyl acetate = 1.5: 1) was used to concentrate the silica gel. Purified by column chromatography on column, 103 mg (yield; 54%) of (3a S , 4 R , 6a R ) -4- (2,6-dichloro-purin-9-yl) -2,2-dimethyl- Tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide was obtained in the oil phase.

¹ H-NMR (CDCl ₃ ) δ: 1.42 (s, 3H), 1.61 (s, 3H), 2.83 (d, 3H), 3.07 (m, 2H), 5.11 (m, 1H), 6.09 (d, 1H ), 7.24 (br s, 1H), 8.52 (s, 1H)

UV (Methanol): λ _max 270nm (pH 7)

Example 11. (3a S ,4 R , 6a R ) -4- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide compound synthesis (11)

(3a S , 4 R , 6a R ) -4- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] in Example 10, supra Ethanol was added to 65 mg (0.16 mmol) of [1,3] dioxol-4-carboxylic acid methylamide, 55 mg (0.20 mmol) of 3-iodobenzylamine hydrochloride, and 0.065 mL (0.46 mmol) of triethylamine. Stir at room temperature for 3 days. The reaction mixture was distilled off under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to obtain 87 mg (yield; 90%) of (3a S , 4 R , 6a). R ) -4- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1, 3] Dioxol-4-carboxylic acid methylamide was obtained in the form of a white foam.

¹ H-NMR (CDCl ₃ ) δ: 1.43 (s, 3H), 1.61 (s, 3H), 2.88 (d, 3H), 3.07 (d, 2H), 4.73 (br d, 2H), 5.09 (m, 1H), 6.11 (d, 1H), 6.49 (br s, 1H), 7.07 (t, 1H), 7.32 (d, 1H), 7.60 (d, 1H), 7.72 (s, 1H), 8.16 (s, 1H)

UV (methanol): λ _max 271 nm (pH 7)

Example 12. (2 R , 3 S ,4 R ) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide) compound synthesis (12)

(3a S, 4 R, 6a R) of Example 11. 4- [2-chloro-6- (3-iodo-benzylamino) - purin-9-yl] -2,2-dimethyl-tetrahydro- To 42 mg (0.07 mmol) of thieno [3,4- d ] [1,3] dioxol-4-carboxylic acid methylamide, 4 ml of an aqueous 80% acetic acid solution was added and stirred at 55 ° C. for 12 hours. After distillation under reduced pressure, the pH was adjusted to neutral by addition of saturated ammonia methanol, and then purified by column chromatography on silica gel using an eluent (dichloromethane and methanol = 7: 1), 20 mg (yield; 52%). of (2 R, 3 S, 4 R) -2- [2- chloro-6- (3-iodo-benzylamino) - purin-9-yl] -3,4-dihydroxy-tetrahydro-thiophen 2-Carboxylic acid methylamide was obtained as a white solid (see Table 1).

Melting Point: 205-207 ℃

[α] ²⁶ _D : -24.5 ( c 0.06, methanol)

υ_max(KBr) / cm^-One: 1066, 1375, 1455, 2924

_{^{1 H-NMR (DMSO- d 6}} ) δ: 2.63 (d, 3H), 2.99 (m, 1H), 3.15 (m, 1H), 4.21 (br s, 1H), 4.73 (br d, 2H), 5.03 (br s, 1H), 5.68 (d, 1H), 6.45 (d, 1H), 7.12 (t, 1H), 7.33 (d, 1H), 7.58 (d, 1H), 7.73 (s, 1H), 8.13 (s, 1H), 8.25 (br s, 1 H), 8.73 (br s, 1 H)

UV (methanol): λ _max 271 nm (pH 7)

(Ⅱ)

Compound group compound R ₁ R ₂ R ₃ R ₃ '' Spectral data Ⅱ 12 3-iodobenzyl Goat Methylaminocarbonyl Hydrogen _{^{1 H-NMR (DMSO- d 6}} ) δ: 2.63 (d, 3H), 2.99 (m, 1H), 3.15 (m, 1H), 4.21 (br s, 1H), 4.73 (br d, 2H), 5.03 (br s, 1H), 5.68 (d, 1H), 6.45 (d, 1H), 7.12 (t, 1H), 7.33 (d, 1H), 7.58 (d, 1H), 7.73 (s, 1H), 8.13 (s, 1H), 8.25 (br s, 1 H), 8.73 (br s, 1 H)

Example 13. ((3a S ,4 R , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-yl) -methanol compound synthesis (13)

(3a S, 4 S, 6a R) -2,2- dimethyl-tetrahydro-thieno [3,4- d] [1,3] dioxole-4-methanol 70 carbaldehyde 5.6g (30.0mmol) Dissolve in mL and add 1.3 g (33.6 mmol) of sodium borohydride in several portions and add at 0 ° C. The reaction mixture was stirred at room temperature for 30 minutes and neutralized with acetic acid. After distillation under reduced pressure, saturated brine was added and extracted with ethyl acetate. The extract obtained by drying and filtering the filtrate was evaporated under reduced pressure. The concentrate obtained was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 2: 1), and purified to 5.5 g (yield; 98%) of ( (3a S, 4 R, 6a R) -2,2- dimethyl-tetrahydro-thieno [3,4- d] [1,3] dioxol-4-yl) methanol was obtained as a white foam .

¹ H-NMR (CDCl ₃ ) δ: 1.33 (s, 3H), 1.53 (s, 3H), 2.41 (br s, 1H), 2.89 (dd, 1H), 3.09 (dd, 1H), 3.44 (dt, 1H), 3.59 (d, 2H), 4.71 (dd, 1H), 4.91 (dt, 1H)

Example 14. Benzoic Acid (3a S ,4 R , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (14)

Of Example 13 ((3a S, 4 R , 6a R) -2,2- dimethyl-tetrahydro-thieno [3,4- d] [1,3] dioxol-4-yl) methanol 2.1 g (11.1 mmol) was dissolved in 20 mL of pyridine and 1.9 g (13.4 mmol) of benzoyl chloride were added at 0 ° C. The reaction mixture was stirred at room temperature for 6 hours, and then 2 ml of methanol was added. The reaction mixture was distilled under reduced pressure, 50 ml of diethyl ether was added thereto, and the resulting solid was filtered. The concentrate obtained by distillation of the filtrate under reduced pressure was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 4: 1), followed by benzoic acid (3a S , 4 R , 6a R ) -2,2-dimethyl 3.2 g (yield; 99%) of tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester were obtained as a colorless oil.

¹ H-NMR (CDCl ₃ ) δ: 1.26 (s, 3H), 1.47 (s, 3H), 2.87 (dd, 1H), 3.09 (dd, 1H), 3.44 (dt, 1H), 4.28 (m, 2H ), 4.72 (dd, 1H), 4.91 (dt, 1H), 7.35-7.99 (m, 5H)

Example 15. Benzoic Acid (3a S ,4 R , 5 R , 6a R )-And (3a S ,4 R , 5 S , 6a R ) -2,2-dimethyl-5-oxo-tetrahydro-5λ ⁴ -Thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (15)

Benzoic acid (3a S , 4 R , 6a R ) -2,2-dimethyl-tetrahydro-thieno [3,

1.4 g (4.6 mmol) of 4- d ] [1,3] dioxol-4-ylmethyl ester were dissolved in 30 mL of dichloromethane, adjusted to -78 ° C, and then 1.0 g (4.6 mmol, 80% of m -chloroperbenzoic acid). A solution of 15 ml of dichloromethane) was added dropwise, and the reaction mixture was stirred at -78 ° C for 45 minutes. After adding saturated sodium bicarbonate water and extracting with dichloromethane, the organic layer was washed with saturated brine, dried and filtered, and the filtrate was distilled under reduced pressure, and the concentrate was purified by eluent (hexane: ethyl acetate = 2: 1). Purification by chromatography, benzoic acid (3a S , 4 R , 5 R , 6a R )-and (3a S , 4 R , 5 S , 6a R ) -2,2-dimethyl-5-oxo-tetrahydro- 3.2 g (yield; 99%) of 5λ ⁴ -thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester were obtained as a solid.

¹ H-NMR (CDCl ₃ ) δ: 1.33 (s, 3H), 1.49 (s, 3H), 3.21 (dd, 1H), 3.37 (dd, 1H), 3.47 (m, 1H), 4.72 (dd, 1H ), 4.87 (dd, 1H), 5.02 (t, 1H), 5.24 (m, 1H), 7.41-8.03 (m, 5H)

Example 16. Benzoic Acid (3a S ,4 R , 6 R , 6a R )-And (3a S ,4 R , 6 S , 6a R ) -6-acetoxy-2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (16)

(3a S , 4 R , 5 R , 6a R )-and (3a S , 4 R , 5 S , 6a R ) -2,2-dimethyl-5-oxo-tetrahydro-5λ ⁴ -in Example 15, above 3.5 g (11.3 mmol) of thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester were dissolved in 90 ml of acetic anhydride and stirred at 100 ° C. for 6 hours, and the reaction mixture was concentrated under reduced pressure. Water was added, extraction was performed with ethyl acetate, and the organic layer was washed with saturated sodium bicarbonate and saturated brine in that order. After drying and filtration of the organic layer, the concentrate obtained by distillation under reduced pressure was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 3: 1) to give benzoic acid (3a S , 4 R , 6 R , 6a). R )-and (3a S , 4 R , 6 S , 6a R ) -6-acetoxy-2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4 2.5 g (yield; 62%) of monomethyl ester were obtained as a syrup.

¹ H-NMR (CDCl ₃ ) δ: 1.30 (s, 3H), 1.50 (s, 3H), 2.03 (s, 3H), 3.76 (dd, 1H), 4.39 (m, 2H), 4.94 (d, 1H ), 4.98 (d, 1H), 6.06 (s, 1H), 7.42-8.06 (m, 5H)

Example 17. (3a S ,4 R , 6 R , 6a R )-And (3a S ,4 R , 6 S , 6a R ) -6- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (17)

415 mg (2.2 mmol) of 2,6-dichloropurine and the amount of ammonium sulfate catalyst were suspended in 80 ml of anhydrous hexamethyldisilazane and stirred under reflux for 6 hours. The reaction mixture was concentrated under reduced pressure to dryness and dissolved in the concentrate in dichloroethane 15㎖ acid dissolved in dichloroethane 20㎖ (3a S, 4 R, 6 R, 6a R) - and (3a S, 4 R, 6 S, 6a R ) -6-acetoxy-2,2-dimethyl-tetrahydro

668 mg (1.9 mmol) of -thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester were added and adjusted to -10 ° C, followed by 0.42 mL (2.18) of trimethylsilyl trifluoromethanesulfonate. mmol) was added and the reaction mixture was stirred at -10 ° C for 20 minutes and stirred at reflux for 4 hours. Saturated sodium bicarbonate was added, stirred for 15 minutes, extracted with dichloromethane, washed with saturated brine, dried, filtered and the filtrate was concentrated under reduced pressure. The resulting concentrate was eluent (hexane: ethyl acetate = (2: 1 in volume). 409 mg (yield; 44%) of benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2,6-dichloro-purine- 9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester with 102 mg (yield; 11%) of benzoic acid (3a S , 4 R , 6 S , 6a R ) -6- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] di Oxol-4-ylmethyl esters were obtained in the form of white foams, respectively.

β-anomer (3a S ,4 R , 6 R , 6a R )

¹ H-NMR (CDCl ₃ ) δ: 1.39 (s, 3H), 1.64 (s, 3H), 4.11 (dt, 1H), 4.57 (dd, 1H), 4.75 (dd, 1H), 5.19 (dd, 1H ), 5.40 (dd, 1H), 6.10 (d, 1H), 7.37-7.97 (m, 5H), 8.38 (s, 1H)

UV (methanol): λ _max 269 nm (pH 7).

α-anomer (3a S ,4 R , 6 S , 6a R )

¹ H-NMR (CDCl ₃ ) δ: 1.37 (s, 3H), 1.64 (s, 3H), 4.08 (m, 1H), 4.51 (dd, 1H), 4.62 (dd, 1H), 5.01 (m, 1H ), 5.11 (m, 1H), 6.50 (d, 1H), 7.37-7.99 (m, 5H), 8.87 (s, 1H)

UV (methanol): λ _max 279 nm (pH 7).

Example 18. Benzoic Acid (3a S ,4 R , 6 R , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (18)

Example 17 acid (3a S, 4 R, 6 R, 6a R) -6- (2,6- dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3, Ethanol in 254 mg (0.53 mmol) of 4- d ] [1,3] dioxol-4-ylmethyl ester, 200 mg (0.68 mmol) of 3-iodobenzylamine hydrochloride, and 0.22 ml (1.53 mmol) of triethylamine 1.5 ml was added and stirred for 3 days at room temperature. The reaction mixture was distilled off under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to give 322 mg (yield; 90%) of benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 1.37 (s, 3H), 1.63 (s, 3H), 4.11 (dt, 1H), 4.57 (dd, 1H), 4.76 (m, 3H), 5.22 (dd, 1H ), 5.38 (dd, 1H), 6.01 (d, 1H), 6.28 (br, 1H), 7.04-8.00 (m, 9H), 8.02 (s, 1H)

UV (methanol): λ _max 272 nm (pH 7).

Example 19. Benzoic Acid (3a S ,4 R , 6 S , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (19)

Benzoic acid (3a S , 4 R , 6 S , 6a R ) -6- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [ 1,3] 80 mg (0.17 mmol) of dioxol-4-ylmethyl ester, 60 mg (0.22 mmol) of 3-iodobenzylamine hydrochloride, and 1.5 ml of ethanol were added to 0.07 mL (0.49 mmol) of triethylamine. Stir at room temperature for 3 days. The reaction mixture was distilled under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to give 102 mg (yield; 90%) of benzoic acid (3a S , 4 R , 6). S , 6a R ) -6- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 1.23 (s, 3H), 1.28 (s, 3H), 3.89 (t, 1H), 3.94 (dd, 1H), 4.08 (dd, 1H), 4.65 (d, 2H ), 5.01 (m 2H), 5.81 (d, 1H), 6.91-7.75 (m, 9H), 8.83 (s, 1H)

UV (methanol): λ _max 282 nm (pH 7).

Example 20. (2 R , 3 R ,4 S , 5 R ) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol compound synthesis (20)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro- To 122 mg (0.18 mmol) of thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester, 6 ml of an aqueous 80% acetic acid solution was added and stirred at 55 ° C. for 12 hours. After distillation under reduced pressure and the reaction mixture, ammonia is tailored to neutral with a saturated methanol solution and then the eluent (dichloromethane: methanol = 15: 1) was purified by column chromatography on silica gel using acid (2 R, 3 S, 4 R , 5 R ) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophen-2-ylmethyl Ester obtained as white foam. The above acid (2 R, 3 S, 4 R, 5 R) -5- [2- chloro-6- (3-iodo-benzylamino) - purin-9-yl] -3,4-dihydroxy Dissolve tetrahydro thiophen-2-ylmethyl ester in 8 ml of methanol, add 30 mg (0.58 mmol) of sodium methoxide, and stir the reaction mixture at room temperature for 4 hours, adjust the liquidity to neutral with acetic acid and distilled under reduced pressure. I was. The concentrate leaching solution (dichloromethane: methanol = 6: 1) was purified by column chromatography on silica gel using 51mg (yield; 53%) of (2 R, 3 R, 4 S, 5 R) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol was obtained as a white solid phase (see Table 3). ).

Melting Point: 112-114 ℃

[α] ²⁶ _D : -28.3 ( c 0.1 methanol)

υ_max(KBr) / cm^-One: 1316, 1622, 3400

_{^{1 H-NMR (DMSO- d 6}} ) δ: 3.29 (m, 1H), 3.62 (m, 1H), 3.78 (m, 1H), 4.20 (d, 1H), 4.61 (m, 3H), 5.17 (t , 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 7.13 (t, 1H), 7.34 (d, 1H), 7.59 (d, 1H), 7.74 (s, 1H), 8.53 (s, 1H), 8.92 (br t, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ42.2, 53.1, 61.1, 62.8, 72.8, 76.7, 126.5, 130.2, 135.3, 135.7, 140.4

UV (methanol): λ _max 273 nm (pH 7).

Example 21. (2 S , 3 R ,4 S , 5 R ) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol compound synthesis (21)

Benzoic acid (3a S , 4 R , 6 S , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro- To 50 mg (0.07 mmol) of thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester was added 3 ml of an aqueous 80% acetic acid solution and stirred at 55 ° C for 12 hours. The reaction mixture was distilled under reduced pressure, the mixture was neutralized with methanol solution saturated with ammonia, and then purified by column chromatography on silica gel using an eluent (dichloromethane: methanol = 15: 1), followed by benzoic acid ( 2R , 3 S , 4 R , 5 S ) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophen-2 Monomethyl ester was obtained as a white foam. Benzoic acid (2 R , 3 S , 4 R , 5 S ) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy -Tetrahydro thiophen-2-ylmethyl ester was dissolved in 4 ml of methanol, and 15 mg (0.29 mmol) of sodium methoxide was added. The reaction mixture was stirred at room temperature for 4 hours, neutralized with acetic acid, and then distilled under reduced pressure. The concentrate leaching solution (dichloromethane: methanol = 6: 1) was purified by column chromatography on silica gel using 21mg (yield; 54%) of (2 S, 3 R, 4 S, 5 R) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol was obtained as a white solid phase (see Table 2). ).

Melting Point: 123-124 ℃

υ_max(KBr) / cm^-One: 1557, 1608, 3429

_{^{1 H-NMR (DMSO- d 6}} ) δ: 3.43 (br d, 1H), 3.56 (dd, 1H), 3.71 (dd, 1H), 4.06 (dd, 1H), 4.27 (dd, 1H), 4.57 ( dd, 1H), 4.69 (dd, 1H), 5.42 (d, 1H), 5.57 (t, 1H), 5.58 (d, 1H), 5.95 (d, 1H), 7.09 (t, 1H), 7.33 (d , 1H), 7.54 (d, 1H), 7.73 (s, 1H), 8.20 (br s, 1H), 8.59 (s, 1H)

UV (methanol): λ _max 283 nm (pH 7).

(Ⅳ)

Compound group compound R ₁ R ₂ R ₃ R ₃ '' Spectral data Ⅳ 21 3-iodobenzyl Goat Hydrogen Hydroxymethyl _{^{1 H-NMR (DMSO- d 6}} ) δ: 3.43 (br d, 1H), 3.56 (dd, 1H), 3.71 (dd, 1H), 4.06 (dd, 1H), 4.27 (dd, 1H), 4.57 ( dd, 1H), 4.69 (dd, 1H), 5.42 (d, 1H), 5.57 (t, 1H), 5.58 (d, 1H), 5.95 (d, 1H), 7.09 (t, 1H), 7.33 (d , 1H), 7.54 (d, 1H), 7.73 (s, 1H), 8.20 (br s, 1H), 8.59 (s, 1H)

Example 22. Benzoic Acid (3a S ,4 R , 6 R , 6a R ) -6- (2-chloro-6-methylamino-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (22)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [ 454 mg (0.94 mmol) of 1,3] dioxol-4-ylmethyl ester was dissolved in 20 mL (40 mmol) of 2 N methylaminetetrahydrofuran, and stirred for 24 hours in a metal sealed container at room temperature. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to give 383 mg (yield; 85%) of benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2-chloro-6-methylamino-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] di Oxol-4-ylmethyl ester was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 1.37 (s, 3H), 1.63 (s, 3H), 3.02 (d, 3H), 4.01 (dt, 1H), 4.57 (dd, 1H), 4.75 (dd, 1H ), 5.23 (dd, 1H), 5.38 (dd, 1H), 6.01 (d, 1H), 6.28 (br s, 1H), 7.40-8.00 (m, 5H), 8.02 (s, 1H)

UV (methanol): λ _max 271 nm (pH 7)

Example 23. (2 R , 3 R ,4 S , 5 R ) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol compound synthesis (23)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2-chloro-6-methylamino-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] 5 ml of an aqueous 80% acetic acid solution was added to 105 mg (0.22 mmol) of [1,3] dioxol-4-ylmethyl ester and stirred at 55 ° C. for 12 hours. After distilling the reaction mixture under reduced pressure, the mixture was neutralized with methanol solution saturated with ammonia, and then purified by column chromatography on silica gel using an eluent (dichloromethane: methanol = 15: 1) to give benzoic acid ( 2R , 3). S, 4 R, 5 R) -5- (2- chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro-thiophen-2-yl-methyl ester is a white foam Was obtained. The above acid (2 R, 3 S, 4 R, 5 R) -5- (2- chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro-thiophen -2 After dissolving the methyl methyl ester in 5 ml of methanol and adding 15 mg (0.29 mmol) of sodium methoxide, the reaction mixture was stirred at room temperature for 4 hours, the mixture was neutralized with acetic acid, and then distilled under reduced pressure. The concentrate leaching solution (dichloromethane: methanol = 6: 1) was purified by column chromatography on silica gel using 38mg; (yield: 52%) (2 R, 3 R, 4 S, 5 R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol was obtained as a white solid phase (see Table 3).

Melting Point: 239-241 ℃

[α] ²⁴ _D : -30.6 ( c 0.1 methanol)

υ_max(KBr) / cm^-One: 1312, 1626, 3419

_{^{1 H-NMR (DMSO- d 6}} ) δ: 2.92 (d, 3H), 3.30 (br s, 1H), 3.62 (m, 1H), 3.79 (m, 1H), 4.20 (dd, 1H), 4.61 ( m, 1H), 5.16 (t, 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 8.26 (br s, 1H), 8.47 (s, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ 27.5, 53.7, 61.7, 63.5, 73.5, 77.4, 118.9, 140.5, 150.0, 153.6, 155.9

UV (methanol): λ _max 271 nm (pH 7)

Example 24. Benzoic Acid (3a S ,4 R , 6 R , 6a R ) -6- (6-amino-2-chloro-purin-9-yl) -2,2-dimethyl

Tetrahydro-thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester compound synthesis (24)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2,6-dichloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [ 454 mg (0.94 mmol) of 1,3] dioxol-4-ylmethyl ester and 25 ml of saturated ammonia ethanol solution were transferred to a metal-sealed container and heated to 50 ° C. for 12 hours. The reaction mixture was concentrated under reduced pressure, and the concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 1: 1) to give 384 mg (yield; 88%) of benzoic acid (3a S , 4 R , 6). R , 6a R ) -6- (6-amino-2-chloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] [1,3] dioxol- 4-ylmethyl ester was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 1.37 (s, 3H), 1.63 (s, 3H), 4.01 (dt, 1H), 4.57 (dd, 1H), 4.77 (dd, 1H), 5.23 (dd, 1H ), 5.38 (dd, 1H), 6.02 (d, 1H), 6.29 (br s, 2H), 7.40-8.00 (m, 5H), 8.02 (s, 1H)

UV (methanol): λ _max 264 nm (pH 7).

Example 25. (2 R , 3 R ,4 S , 5 R ) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol compound synthesis (25)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (6-amino-2-chloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] 5 ml of an aqueous 80% acetic acid solution was added into 102 mg (0.22 mmol) of [1,3] dioxol-4-ylmethyl ester, followed by stirring at 55 ° C for 12 hours. After distilling the reaction mixture under reduced pressure, the mixture was neutralized with methanol solution saturated with ammonia, and then purified by column chromatography on silica gel using an eluent (dichloromethane: methanol = 15: 1) to give benzoic acid ( 2R , 3). S, 4 R, 5 R) -5- (6- amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro-thiophen-4-yl methyl ester a white foam as the Obtained. Benzoic acid (2 R , 3 S , 4 R , 5 R ) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophen-4- After dissolving the monomethyl ester in 5 ml of methanol and adding 15 mg (0.29 mmol) of sodium methoxide, the reaction mixture was stirred at room temperature for 4 hours, the mixture was neutralized with acetic acid, and then distilled under reduced pressure. The concentrate leaching solution (dichloromethane: methanol = 6: 1) was purified by column chromatography on silica gel using 37mg (yield; 53%) of (2 R, 3 R, 4 S, 5 R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol was obtained as a white solid phase (see Table 3).

Melting Point: 220-222 ℃

[α] ²⁶ _D : -24.6 ( c 0.07 methanol)

υ_max(KBr) / cm^-One: 1204, 1647, 3420

_{^{1 H-NMR (DMSO- d 6}} ) δ: 3.30 (br s, 1H), 3.62 (m, 1H), 3.79 (m, 1H), 4.20 (dd, 1H), 4.61 (dd, 1H), 5.16 ( t, 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 7.81 (br s, 2H), 8.48 (s, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ 53.3, 61.2, 63.1, 73.1, 76.9, 118.0, 140.4, 150.7, 152.9, 156.7

UV (methanol): λ _max 264 nm (pH 7)

Example 26. Benzoic Acid (2 R , 3 S ,4 R , 5 R ) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophen-4-ylmethyl ester compound synthesis (26)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (6-amino-2-chloro-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] 5 ml of 80% acetic acid solution was added to 102 mg (0.22 mmol) of [1,3] dioxol-4-ylmethyl ester and stirred at 55 ° C. for 12 hours. The reaction mixture was distilled under reduced pressure, and ammonia saturated methanol was added. Align the liquid to neutral with a solution eluent (dichloromethane: methanol = 15: 1) was purified by column chromatography 63mg on silica gel using; acid (2 R, (yield 68%) 3 S, 4 R , 5 R ) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophen-4-ylmethyl ester was obtained in white foam.

Example 27. (2 R , 3 S ,4 R , 5 R )-[5- (6-amino-2-chloro-purin-9-yl) -3,4-bis- ( tert -Butyl-dimethyl-silanyloxy) -tetrahydro thiophen-2-yl] -methanol compound synthesis (27)

Example 26 acid (2 R, 3 S, 4 R, 5 R) -5- (6- amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro-thiophen 230 mg (0.55 mmol) of -4-ylmethyl ester, 225 mg (3.30 mmol) of imidazole, and 249 mg (1.65 mmol) of tert -butylchlorodimethylsilane were dissolved in 10 ml of anhydrous N , N -dimethylformamide, followed by 24 hours at 50 ° C. After stirring, water was added to the reaction mixture, followed by extraction with dichloromethane. The organic layer was washed with water, saturated sodium bicarbonate water and saturated brine in that order, dried, filtered and distilled under reduced pressure. Dissolve in mL and add 45 mg (0.83 mmol) of sodium methoxide. The reaction mixture was stirred at room temperature for 4 hours, neutralized with acetic acid, and distilled under reduced pressure. The concentrate was purified by column chromatography on silica gel using an eluent (hexane: ethyl acetate = 2: 1) to give 169 mg ( yield; 56%) of (2 R, 3 S, 4 R, 5 R) - [5- (6- amino-2-chloro-purin-9-yl) -3,4-bis - (tert - butyl - Dimethyl-silanyloxy) -tetrahydro thiophen-2-yl] -methanol was obtained in white foam.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.79 (s, 9H), 0.83 (s, 9H), 3.41 (dd, 1H), 4.21 (m, 1H), 4.56 (dd, 1H ), 4.73 (m, 1H), 4.86 (dd, 1H), 5.71 (d, 1H), 6.20 (br s, 1H), 7.61 (br s, 2H), 8.12 (s, 1H)

UV (methanol): λ _max 263 nm (pH 7)

Example 28. (2 S , 3 S ,4 R , 5 R ) -5- (6-amino-2-chloro-purin-9-yl) -3,4-bis- ( tert -Butyl

-Dimethyl-silanyloxy) -tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis (28)

Example 27 (2 R, 3 S, 4 R, 5 R) - [5- (6- Amino-2-chloro-purin-9-yl) -3,4-bis - (tert-butyl-dimethyl- 150 mg (0.27 mmol) of -silanyloxy) -tetrahydro thiophen-2-yl] -methanol was dissolved in 20 ml of anhydrous N , N -dimethylformamide, followed by addition of 2.34 g (6.21 mmol) of pyridinium dichromate, followed by reaction. The mixture was stirred at room temperature for 20 hours. Water was added to the reaction mixture, followed by extraction with ethyl acetate several times. The organic layer was dried, filtered and the filtrate was distilled under reduced pressure to obtain a concentrate, an acid derivative. The concentrate was dissolved in 6 ml of acetone without further purification, 1.0 ml (10.57 mmol) of dimethyl sulfate and 100 mg (0.72 mmol) of potassium carbonate were added and stirred at room temperature for 2 hours, and the reaction mixture was concentrated under reduced pressure. Water was dissolved in ethyl acetate and washed with water and brine. The organic layer was distilled under reduced pressure to obtain a methyl ester derivative in the form of a foam. 20 ml (40 mmol) of 2 N methylaminetetrahydrofuran solution was added to the methyl ester derivative without further purification. The reaction was carried out for 24 hours. After distillation under reduced pressure and the reaction mixture, and the resulting concentrate the eluate (hexane: ethyl acetate = 1: 1) was purified by column chromatography 80mg on silica gel; (2 S, (yield 51%) 3 S , 4 R, 5 R) -5- (6- amino-2-chloro-purin-9-yl) -3,4-bis - (tert-butyl-dimethyl-silanyloxy) -2-tetrahydro-thiophen Carboxylic acid methylamide was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.79 (s, 9H), 0.85 (s, 9H), 2.67 (d, 3H), 3.78 (d, 1H), 4.35 (m, 1H ), 4.57 (m, 1H), 5.70 (d, 1H), 7.72 (br s, 2H), 7.72 (br s, 1H), 8.10 (s, 1H)

UV (methanol): λ _max 264 nm (pH 7)

Example 29. (2 S , 3 S ,4 R , 5 R ) -5- (6-Amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis (29)

Example 28 (2 S, 3 S, 4 R, 5 R) -5- (6- amino-2-chloro-purin-9-yl) -3,4-bis

140 mg (0.24 mmol) of (( tert -butyl-dimethyl-silanyloxy) -tetrahydro thiophene-2-carboxylic acid methylamide were dissolved in 10 ml of anhydrous tetrahydrofuran and 0.67 ml (0.67 mmol, 1M) of tetrabutylammonium fluoride. Tetrahydrofuran solution) was added and the reaction mixture was stirred at room temperature for 1 hour. Was evaporated under reduced pressure and the reaction mixture and concentrate the eluent (dichloromethane: methanol = 4: 1) was purified by column chromatography 58mg on silica gel; (2 S, (yield 69%) 3 S, 4 R , 5 R ) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide was obtained as a white solid (Table 3). Reference).

Melting Point: 233-235 ℃

[α] ²⁶ _D : -20.2 ( c 0.1 methanol)

_{^{1 H-NMR (DMSO- d 6}} ) δ: 2.70 (d, 3H), 3.82 (d, 1H), 4.36 (dd, 1H), 4.53 (m, 1H), 5.60 (d, 1H), 5.78 (d , 1H), 5.82 (d, 1H), 7.87 (br s, 2H), 8.33 (br d, 1H), 8.55 (s, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ 51.8, 57.5, 62.5, 75.4, 78.1, 118.4, 140.1, 150.5, 153.0, 156.7, 170.7

UV (methanol): λ _max 264 nm (pH 7)

Example 30. Benzoic Acid (2 R , 3 S ,4 R , 5 R ) -5- (2-Chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophen-2-ylmethyl ester compound synthesis (30)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- (2-chloro-6-methylamino-purin-9-yl) -2,2-dimethyl-tetrahydro-thieno [3,4- d ] 5 ml of 80% acetic acid aqueous solution was added to 105 mg (0.22 mmol) of [1,3] dioxol-4-ylmethyl ester and stirred at 55 ° C. for 12 hours. After distilling the reaction mixture under reduced pressure, the solution was neutralized with methanol solution saturated with ammonia, and then purified by column chromatography on silica gel using an eluent (dichloromethane: methanol = 15: 1), and 93 mg (yield; 68). %) benzoic acid (2 R, 3 S, 4 R, 5 R) -5- (2- chloro-6-methyl-amino-purin-9-yl) -3,4-dihydroxy-tetrahydro-thiophen- 2-ylmethyl ester was obtained as a white foam.

Example 31. (2 R , 3 S ,4 R , 5 R )-[3,4-bis- ( tert -Butyl-dimethyl-silanyloxy) -5- (2-chloro

-6-Methylamino-purin-9-yl) -tetrahydro thiophen-2-yl] -methanol compound synthesis (31)

Example 30 acid (2 R, 3 S, 4 R, 5 R) of 5- (2-chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro-T 160 mg (0.38 mmol) of fen-2-ylmethyl ester, 155 mg (2.28 mmol) of imidazole, and 172 mg (1.14 mmol) of tert -butylchlorodimethylsilane were dissolved in 10 ml of anhydrous N , N -dimethylformamide and 24 at 50 ° C. Stirred for time. Water was added to the reaction mixture, followed by extraction with dichloromethane. The organic layer was washed with water, saturated sodium bicarbonate and saturated brine in that order, dried, filtered and the filtrate was distilled under reduced pressure. The concentrate was dissolved in 8 ml of methanol without any further purification. 45 mg (0.83 mmol) of sodium methoxide were added. The reaction mixture was stirred at room temperature for 4 hours, neutralized with acetic acid and distilled under reduced pressure. The concentrate was purified by column chromatography on silica gel using eluent (hexane: ethyl acetate = 2: 1) to give 112 mg (yield; 53%) of (2 R , 3 S , 4 R , 5 R )-[3 , 4-bis- ( tert -butyl-dimethyl-silanyloxy) -5- (2-chloro-6-methylamino-purin-9-yl) -tetrahydro thiophen-2-yl] -methanol white foam Obtained as a phase.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.81 (s, 9H), 0.86 (s, 9H), 3.13 (br s, 3H), 3.72 (dd, 1H), 4.24 (m, 1H), 4.48 (dd, 1H), 4.75 (m, 1H), 4.87 (dd, 1H), 5.67 (d, 1H), 5.88 (br s, 1H), 8.00 (s, 1H), 8.03 (br s , 1H)

UV (methanol): λ _max 269 nm (pH 7)

Example 32. (2 S , 3 S ,4 R , 5 R ) -3,4-bis- ( tert -Butyl-dimethyl-silanyloxy) -5- (2-chloro-6-methylamino-purin-9-yl) -tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis (32)

In Example 31 (2 R, 3 S, 4 R, 5 R) - [3,4- bis - (tert-butyl-dimethyl-silanyloxy) -5- (2-chloro-6-methylamino- Dissolve 105 mg (0.19 mmol) of Purin-9-yl) -tetrahydro thiophen-2-yl] -methanol in 15 ml of anhydrous N , N -dimethylformamide and add 1.64 g (4.36 mmol) of pyridinium dichromate. The reaction mixture was stirred at room temperature for 20 hours. Water was added to the reaction mixture and extracted with ethyl acetate several times. The organic layer was dried and filtered to dry the filtrate under reduced pressure to obtain an acid derivative. The concentrate was dissolved in 4 ml of acetone without further purification. 1.0 ml of sulfate (10.57 mmol) and 80 mg (10.58 mmol) of potassium carbonate were added and stirred at room temperature for 2 hours. The reaction mixture was distilled under reduced pressure, and the resulting concentrate was dissolved in ethyl acetate and washed with water and brine. The organic layer was distilled under reduced pressure to obtain a methyl ester derivative in the form of a foam. 15 ml (30 mmol) of 2 N methylaminetetrahydrofuran solution was added to the methyl ester derivative without further purification. The reaction was carried out for 24 hours. After distillation under reduced pressure and the reaction mixture and the resulting concentrate the eluate (hexane: ethyl acetate = 1: 1) was purified by column chromatography 48mg (yield; 44%) on silica gel using a (2 S, 3 S, 4 R, 5 R) -3,4- bis - (tert-butyl-dimethyl-silanyloxy) -5- (2-chloro-6-methylamino-purin-9-yl) -tetrahydro-thiophen -2 Carboxylic acid methylamide was obtained as a white foam.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.82 (s, 9H), 0.85 (s, 9H), 3.21 (br s, 3H), 3.21 (br s, 3H), 3.75 (d , 1H), 4.28 (m, 1H), 4.65 (m, 1H), 5.77 (d, 1H), 7.12 (br s, 1H), 8.03 (br s, 1H), 8.44 (s, 1H)

UV (methanol): λ _max 270 nm (pH 7)

Example 33. (2 S , 3 S ,4 R , 5 R ) -5- (2-Chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis (33)

(2 S, 3 S, 4 R, 5 R) -3,4- bis - (tert-butyl-dimethyl-silanyloxy) -5- (2-chloro-6-methylamino-purin-9-yl) 48 mg (0.08 mmol) of tetrahydro thiophene-2-carboxylic acid methylamide were dissolved in anhydrous tetrahydrofuran, 0.22 ml (0.22 mmol, 1 M tetrahydrofuran solution) of tetrabutylammonium fluoride was added, and the reaction mixture was allowed to stand at room temperature. Stir for hours. It was evaporated under reduced pressure and the reaction mixture and concentrate the eluent (dichloromethane: methanol = 6: 1) was purified by column chromatography on silica gel using, 19mg; (2 S, (yield 65%) 3 S, 4 R , 5R ) -5- (2-chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide was obtained as a white solid phase ( See Table 3).

Melting Point: 139-141 ℃

[α] ¹⁹ _D : -22.0 ( c 0.13 methanol)

_{^{1 H-NMR (DMSO- d 6}} ) δ: 2.69 (d, 3H), 2.92 (d, 3H), 3.81 (d, 1H), 4.36 (dd, 1H), 4.52 (dd, 1H), 5.61 (d , 1H), 5.78 (d, 1H), 5.82 (d, 1H), 7.23 (br s, 1H), 8.33 (br d, 2H), 8.54 (s, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ 51.8, 55.5, 62.6, 69.6, 75.4, 78.2, 118.5, 139.9, 149.5, 153.3, 155.5, 170.7

UV (methanol): λ _max 270 nm (pH 7)

Example 34. (2 R , 3 S ,4 R , 5 R )-(3,4-bis- ( tert -Butyl-dimethyl-silanyloxy) -5- [2-chloro

-6- (3-iodo-benzylamino) -purin-9-yl] -tetrahydro thiophen-2-yl} -methanol compound synthesis (34)

Benzoic acid (3a S , 4 R , 6 R , 6a R ) -6- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -2,2-dimethyl-tetrahydro- To 122 mg (0.18 mmol) of thieno [3,4- d ] [1,3] dioxol-4-ylmethyl ester, 6 ml of an aqueous 80% acetic acid solution was added and stirred at 55 ° C. for 12 hours. The reaction mixture was distilled under reduced pressure, the mixture was neutralized with methanol solution saturated with ammonia, and then purified by column chromatography on silica gel using an eluent (dichloromethane: methanol = 15: 1), followed by benzoic acid ( 2R , 3 S , 4 R , 5 R ) -5- [2-chloro-6- (iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophen-2-yl Methyl esters were obtained as white foam. Acid obtained above (2 R, 3 S, 4 R, 5 R) -5- [2- chloro-6- (iodo-benzylamino) - purin-9-yl] -3,4-dihydroxy-tetrahydro 450 mg (0.71 mmol) of hydrothiophen-2-ylmethyl ester, 286 mg (4.20 mmol) of imidazole and 316 mg (2.10 mmol) of tert -butylchlorodimethylsilane were dissolved in 20 ml of anhydrous N , N -dimethylformamide, Stir for 24 hours. Water was added to the reaction mixture, followed by extraction with dichloromethane. The organic layer was washed with water, saturated sodium bicarbonate and saturated brine in that order, dried, filtered and distilled under reduced pressure. The concentrate was taken up in 15 ml of methanol without further purification and 90 mg (1.74 mmol) of sodium methoxide were added. The reaction mixture was stirred at room temperature for 4 hours, neutralized with acetic acid and distilled under reduced pressure. The concentrate eluate (hexane: ethyl acetate = 2: 1) was purified by column chromatography 290mg on silica gel using; (yield: 54%) (2 R, 3 S, 4 R, 5 R) - {3 , 4-bis- ( tert -butyl-dimethyl-silanyloxy) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -tetrahydro thiophen-2- Il} -methanol was obtained as a white solid phase.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.80 (s, 9H), 0.85 (s, 9H), 3.29 (dd, 1H), 4.24 (m, 1H), 4.47 (dd, 1H ), 4.73 (m, 3H), 4.86 (dd, 1H), 5.67 (d, 1H), 6.15 (br s, 1H), 7.02 (t, 1H), 7.27 (d, 1H), 7.43 (d, 1H ), 7.59 (br s, 1H), 7.67 (s, 1H), 8.03 (s, 1H)

UV (methanol): λ _max 272 nm (pH 7)

Example 35. (2 S , 3 S ,4 R , 5 R ) -3,4-bis- ( tert -Butyl-dimethyl-silanyloxy) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis ( 35)

(2 R, 3 S, 4 R, 5 R) - {3,4- bis - (tert-butyl-dimethyl-silanyloxy) -5- [2-chloro-6- (3-iodo-benzylamino 228 mg (0.30 mmol) of) -purin-9-yl] -tetrahydro thiophen-2-yl} -methanol are dissolved in 20 ml of anhydrous N , N -dimethylformamide and 2.6 g (7.0 mmol) of pyridinium dichromate are added. The reaction mixture was then stirred at room temperature for 20 hours. Water was added to the reaction mixture and extracted with ethyl acetate several times. The organic layer was dried and filtered to distill the filtrate under reduced pressure to obtain a concentrate as an acid derivative. The concentrate was dissolved in 6 ml of acetone without further purification. 1.0 ml of sulfate (10.57 mmol) and 100 mg (0.72 mmol) of potassium carbonate were added and stirred at room temperature for 2 hours. The reaction mixture was distilled under reduced pressure, and the resulting concentrate was dissolved in ethyl acetate and washed with water and brine. The organic layer was distilled under reduced pressure to obtain a methyl ester derivative in the form of a bubble, and 20 ml (40 mmol) of 2 N methylaminetetrahydrofuran solution was added to the methyl ester derivative without further purification. The reaction was time. After distillation under reduced pressure and the reaction mixture and the resulting concentrate the eluate (hexane: ethyl acetate = 1: 2) using a mixed solution of purified by column chromatography on silica gel, 99mg; (yield: 42%) (2 S, 3 S, 4 R, 5 R ) -3,4- bis - (tert-butyl-dimethyl-silanyloxy) -5- [2-chloro-6- (3-iodo-benzylamino) - purin -9 -Yl] -tetrahydro thiophene-2-carboxylic acid methylamide was obtained in the form of white foam.

¹ H-NMR (CDCl ₃ ) δ: 0.01 (m, 12H), 0.70 (s, 9H), 0.84 (s, 9H), 2.70 (d, 3H), 3.81 (d, 1H), 4.39 (m, 1H ), 4.55 (m, 1H), 4.61 (d, 2H), 5.83 (d, 1H), 7.15 (t, 1H), 7.36 (d, 1H), 7.61 (d, 1H), 7.76 (s, 1H) , 8.33 (br s, 1 H), 8.61 (d, 1 H), 9.00 (br s, 1 H)

UV (methanol): λ _max 272 nm (pH 7)

Example 36. (2 S , 3 S ,4 R , 5 R ) -5- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide compound synthesis ( 36)

Example 35 (2 S, 3 S, 4 R, 5 R) -3,4- bis-5- [2-chloro-6- (3-iodo (silanyloxy tert-butyl-dimethyl) 75 mg (0.09 mmol) of do-benzylamino) -purin-9-yl] -tetrahydro thiophene-2-carboxylic acid methylamide was dissolved in 5 ml of anhydrous tetrahydrofuran and 0.25 ml (0.25 mmol, 1 mol) of tetrabutylammonium fluoride. Tetrahydrofuran solution) was added and the reaction mixture was stirred at room temperature for 1 hour. It was evaporated under reduced pressure and the reaction mixture and concentrate the eluent (dichloromethane: methanol = 7: 1) was purified by column chromatography on silica gel using, 33mg; (2 S, (yield 63%) 3 S, 4 R , 5 R ) -5- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide Was obtained as a white solid phase (see Table 3).

Melting Point: 140-141 ℃

[α] 25 _D : -19.5 ( c 0.32 methanol)

_{^{1 H-NMR (DMSO- d 6}} ) δ: 2.69 (d, 3H), 3.82 (d, 1H), 4.37 (dd, 1H), 4.55 (m, 1H), 4.60 (d, 2H), 5.59 (d , 1H), 5.78 (d, 1H), 5.83 (d, 1H), 7.13 (t, 1H), 7.35 (d, 1H), 7.60 (d, 1H), 7.75 (s, 1H), 8.32 (br d , 1H), 8.60 (s, 1H), 8.99 (br t, 1H)

¹³ C-NMR (DMSO- d ₆ ): δ 42.5, 51.8, 59.7, 62.6, 75.4, 78.2, 118.4, 126.8, 130.5, 135.5, 136.0, 140.3, 141.8, 149.9, 153.0, 154.7, 170.3, 170.7 UV (methanol ): λ _max 272nm (pH 7)

(Ⅲ)

Compound group compound R ₁ R ₂ R ₃ R ₃ '' Spectral data Ⅲ 20 3-iodobenzyl Goat Hydrogen Hydroxymethyl ¹ H-NMR (DMSO-d ₆ ) δ: 3.29 (m, 1H), 3.62 (m, 1H), 3.78 (m, 1H), 4.20 (d, 1H), 4.61 (m, 3H), 5.17 (t , 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 7.13 (t, 1H), 7.34 (d, 1H), 7.59 (d, 1H), 7.74 (s, 1H), 8.53 (s, 1H), 8.92 (brt, 1H) 23 methyl Goat Hydrogen Hydroxymethyl ¹ H-NMR (DMSO-d ₆ ) δ: 2.92 (d, 3H), 3.30 (br s, 1H), 3.62 (m, 1H), 3.79 (m, 1H), 4.20 (dd, 1H), 4.61 ( m, 1H), 5.16 (t, 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 8.26 (br s, 1H), 8.47 (s, 1H) 25 Hydrogen Goat Hydrogen Hydroxymethyl ¹ H-NMR (DMSO-d ₆ ) δ: 3.30 (br s, 1H), 3.62 (m, 1H), 3.79 (m, 1H), 4.20 (dd, 1H), 4.61 (dd, 1H), 5.16 ( t, 1H), 5.34 (d, 1H), 5.58 (d, 1H), 5.77 (d, 1H), 7.81 (br s, 2H), 8.48 (s, 1H)

Compound group compound R ₁ R ₂ R ₃ R ₃ '' Spectral data Ⅲ 29 Hydrogen Goat Hydrogen Methylaminocarbonyl ¹ H-NMR (DMSO-d ₆ ) δ: 2.70 (d, 3H), 3.82 (d, 1H), 4.36 (dd, 1H), 4.53 (m, 1H), 5.60 (d, 1H), 5.78 (d , 1H), 5.82 (d, 1H), 7.87 (br s, 2H), 8.33 (br d, 1H), 8.55 (s, 1H) 33 methyl Goat Hydrogen Methylaminocarbonyl ¹ H-NMR (DMSO-d ₆ ) δ: 2.69 (d, 3H), 2.92 (d, 3H), 3.81 (d, 1H), 4.36 (dd, 1H), 4.52 (dd, 1H), 5.61 (d , 1H), 5.78 (d, 1H), 5.82 (d, 1H), 7.23 (br s, 1H), 8.33 (br d, 2H), 8.54 (s, 1H) 36 3-iodobenzyl Goat Hydrogen Methylaminocarbonyl ¹ H-NMR (DMSO-d ₆ ) δ: 2.69 (d, 3H), 3.82 (d, 1H), 4.37 (dd, 1H), 4.55 (m, 1H), 4.60 (d, 2H), 5.59 (d , 1H), 5.78 (d, 1H), 5.83 (d, 1H), 7.13 (t, 1H), 7.35 (d, 1H), 7.60 (d, 1H), 7.75 (s, 1H), 8.32 (br d , 1H), 8.60 (s, 1H), 8.99 (br t, 1H)

Experimental Example 1.Binding affinity experiment of 4-thio adenosine derivative compound

Cell culture and receptor binding

Chinese Hamster Ovary (CHO, ATCC; US Cell Line Bank No. CCL-61) cells expressing A ₃ receptors were treated with 10% fetal bovine serum (FBS) and penicillin / streptomycin (100 units / ml and 100 μg). / Ml) was cultured under the conditions of 37 ℃, 5% carbon dioxide in F-12 (Gibco, USA) medium.

Binding affinity measurement of [[ ¹²⁵ I] -4-amino-3-iodobenzyl] adenosine-5'-N-methyluronamid ([ ¹²⁵ I] AB-MECA) to CHO cell membranes was determined using 50 μl of [ ¹²⁵ I] AB-MECA], 100 μl membrane float, 50 μl buffer solution containing 50 μl of inhibitor. The inhibitor was first dissolved in dimethylsulfoxide (DMSO) and then diluted with buffer. Care should be taken to ensure that the final concentration of DMSO does not exceed 1%, incubate for 1 hour at 37 ° C, then use a cell collector (TOMTEC, USA) and quickly through a GF / B (Whatman, USA) filter. Filtered. The test tubes were washed three times with 3 ml of buffer and radioactivity was determined using a γ-counter. Nonspecific binding is determined in the presence of 40 μM R-PIA, and the equilibrium constant Ki value is Cheng-Prusoff under the assumption that the K _d value of [ ¹²⁵ I] AB-MECA is 1.48 nM. ) Was determined based on the equation.

[ ³ H] PIA ((R) -N ^6- (phenylisopropyl) adenosine) to A ₁ and [ ³ H] CGS 21680 (2-[[[4- (2-carboxyethyl) phenyl] ethyl Measurement of binding of amino] -5'-N-ethylcarbamoyl) adenosine) to A _2a is performed as follows, and adenosine deaminase is a radioactive ligand when incubating the brain membrane for 30 minutes at 30 ° C. And put together during incubation, and set IC ₅₀ values for individual compounds at at least six different concentrations, and K _i values were determined using a float program. At this time, the K _d values of [ ³ H] PIA and [ ³ H] CGS 21680 were determined based on the V- Prusov equation under the assumption that the K _d values were 1.0 and 14 nM, respectively.

Of the known adenosine derivatives, CI-IB-MECA with the control group is very for the adenosine rat brain A _1, A ₂ and A _1, A _2, while showing an excellent efficacy (agonist) activity against A ₃ from A ₃ It is known to show low affinity (Hea O. Kim, et al . ; J. Med. Chem. , 37 (21), pp 364-3621, 1994).

As a result of the experiment, among the novel thionucleoside compounds of the present invention, the compounds of the general formula (III) exhibited a K _i of 0.28 to 4.9 nM with respect to the adenosine A ₃ receptor, thus exhibiting potent efficacy compared to the known human A ₃ receptor. agonist), and compounds of formula (II) exhibited an excellent antagonist action with K _i of 4.3 to 8.0nM, while much lower affinity than other compounds for other receptors A ₁ and A ₂ . Indicated. In particular, compound 33 is the most potent and most selective A ₃ agonist among the known compounds.

As a result, it was confirmed that the present compounds because they show an excellent selectivity for the adenosine A ₃ receptor, adenosine A ₃ can be effectively used in cancer and inflammatory diseases caused by receptor activation.

(Ⅴ)

compound Compound group R ₁ R ₃ R ₃ '' Ki (nM) or% hA ₁ rA ₁ hA _2A rA _2A hA ₃ Cl-IB-MECA 3-iodobenzyl Hydrogen CONHMe 1240 ± 320 820 ± 570 5360 ± 2470 470 ± 365 1.0 ± 0.2 20 Ⅲ 3-iodobenzyl Hydrogen CH ₂ OH 127 ± 35 285 ± 108 3.2 ± 0.9 25 Ⅲ Hydrogen Hydrogen CH ₂ OH 238 ± 107 705 ± 184 4.9 ± 1.3 23 Ⅲ methyl Hydrogen CH ₂ OH 780 ± 280 <10% (10 μM) 0.8 ± 0.1 12 Ⅱ 3-iodobenzyl CONHMe Hydrogen 110 ± 12 188 ± 40 50% (10 μM) <10% (10 μM) 4.3 ± 1.2 33 Ⅲ methyl Hydrogen CONHMe 1330 ± 242 198 ± 14 20% (10 μM) 6340 ± 90 0.28 ± 0.09 36 Ⅲ 3-iodobenzyl Hydrogen CONHMe 193 ± 46 140 ± 43 223 ± 36 348 ± 110 0.38 ± 0.07 29 Ⅲ Hydrogen Hydrogen CONHMe 89.2 ± 11.7 294 ± 115 158 ± 29 <10% (10 μM) 0.40 ± 0.06

Experimental Example 2. Toxicity Test

In order to test the toxicity of the compounds prepared herein, animal experiments were performed.

The compound of the present invention was divided into three groups of 25 ± 5g of ICR mice (central laboratory animals) and 235 ± 10g of SPF Sprague Dawley rats. Were intraperitoneally administered at doses of 20 mg / kg, 10 mg / kg and 1 mg / kg, respectively, and observed for 24 hours.

As a result, no deaths were observed in all three groups, and no symptoms were observed in weight gain and feed intake, and therefore, the thionucleoside derivative compound was found to be a safe drug. .

Thionucleoside derivative compounds of the present invention can be administered in the following formulations, the formulation examples below are merely to illustrate the invention, thereby not limiting the content of the invention.

Formulation Example 1 Preparation of Powder

500 mg of dry powder of compound 33

Corn Starch 100mg

Lactose 100mg

Talc 10mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

100 mg of dry powder of compound 33

Corn Starch 100mg

Lactose 100mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

50 mg of dry powder of compound 33

Lactose 50mg

1 mg magnesium stearate

The above ingredients are mixed and compressed into tablets according to a conventional method for preparing capsules to fill gelatin capsules.

Formulation Example 4 Preparation of Injection

10 mg of dry powder of compound 33

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injectable drug, the active ingredient is dissolved in distilled water for injection, the pH is adjusted to about 7.5, and the whole is filled with 2 ml of ampoules with injectable distilled water for sterilization.

Formulation Example 5 Preparation of Liquid

1 g dry powder of compound 33

10 g per isomerization

10g per book

Lemon flavor

Purified water

According to the conventional method for preparing a liquid solution, each component is added to the purified water, dissolved, and lemon flavor is added, and then purified water is added to adjust the total amount to 100 ml, and then filled into a brown bottle to prepare a liquid solution.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

Since the novel thionucleoside derivative compound of the present invention and the pharmaceutical composition containing the same have selective activity on the adenosine A ₃ receptor, it can be usefully used as an effective medicine for preventing and treating various cancers or inflammatory bowel disease and inflammatory disease. Can be.

Claims (12)

Compound represented by the following general formula (I) or a pharmaceutically acceptable salt or isomer thereof: (Ⅰ) In the above formula, X is a sulfur atom or an oxygen atom; R ₁ is a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, a benzyl group, a halobenzyl, or a phenylalkyl group; R ₂ is a hydrogen atom, a halogen group, an alkoxy group, an alkenyl group, an alkynyl group, an alkylthio group or a thi group; R ₃ and R ₃ ′ are hydroxyalkyl groups, alkoxycarbonyl groups or alkylaminocarbonyl groups of 1 to 5 carbon atoms, provided that R ₃ and R ₃ ′ do not have the same substituent at the same time; R ₄ is a hydrogen atom or an alkyl group having 1 to 5 carbon atoms. The compound or isomer of formula (II) according to claim 1, wherein R ₃ ′ and R ₄ are hydrogen atoms and X is a sulfur atom. (Ⅱ) In the above formula, R ₁ is a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, benzyl group, halobenzyl, phenylalkyl group, R ₂ is a hydrogen atom, a halogen group, an alkoxy group, an alkenyl group, an alkynyl group, an alkylthio group or a thi group, R <_3> is a C1-C5 hydroxyalkyl group, an alkoxycarbonyl group, or a C1-C5 alkylaminocarbonyl group. The compound according to claim 2, wherein (2R, 3S, 4R) -2- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro A compound that is thiophene-2-carboxylic acid methylamide. The compound or isomer of formula (III) according to claim 1, wherein R ₃ and R ₄ are hydrogen atoms and X is a sulfur atom. (Ⅲ) In the above formula, R ₁ is a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, benzyl group, halobenzyl, phenylalkyl group, R ₂ is a hydrogen atom, a halogen group, an alkoxy group, an alkenyl group, an alkynyl group, an alkylthio group or a thi group, R <_3>'is a C1-C5 hydroxyalkyl group, an alkoxycarbonyl group, or a C1-C5 alkylaminocarbonyl group. The method of claim 4, wherein (2R, 3R, 4S, 5R) -2- [2-Chloro-6- (3-iodo-benzylamino) -purin-9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4 -Dior, (2R, 3R, 4S, 5R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol, (2R, 3R, 4S, 5R) -2- (2-Chloro-6-methylamino-purin-9-yl) -5-hydroxymethyl-tetrahydro thiophene-3,4-diol, (2S, 3S, 4R, 5R) -5- (6-amino-2-chloro-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide, (2S, 3S, 4R, 5R) -5- (2-Chloro-6-methylamino-purin-9-yl) -3,4-dihydroxy-tetrahydro thiophene-2-carboxylic acid methylamide, (2S, 3S, 4R, 5R) -5- [2-chloro-6- (3-iodo-benzylamino) -purin-9-yl] -3,4-dihydroxy-tetrahydro thiophene-2 The compound is carboxylic acid methylamide. A compound according to claim 1, wherein R ₃ and R ₄ are hydrogen atoms and X is a sulfur atom or a compound of the general formula (IV). (Ⅳ) In the above formula, R ₁ is a hydrogen atom, an alkyl group having 1 to 5 carbon atoms, benzyl group, halobenzyl, phenylalkyl group, R ₂ is a hydrogen atom, a halogen group, an alkoxy group, an alkenyl group, an alkynyl group, an alkylthio group or a thi group, R <_3>'is a C1-C5 hydroxyalkyl group, an alkoxycarbonyl group, or a C1-C5 alkylaminocarbonyl group. The compound according to claim 6, wherein (2S, 3R, 4S, 5R) -2- [2-chloro-6- (3-iodo-benzylamino) -purine -9-yl] -5-hydroxymethyl-tetrahydro thiophene-3,4-diol. A pharmaceutical composition having adenosine A ₃ receptor selection activity comprising the general formula compound of any one of claims 1 to 7 as an active ingredient and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 8, which is effective for the prevention and treatment of cancer or inflammatory diseases by adenosine A ₃ receptor selection activity. A pharmaceutical composition for cancer diseases comprising the general formula compound of any one of claims 1 to 7 as an active ingredient and a pharmaceutically acceptable carrier. The method of claim 10, wherein the cancer is lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, anal muscle cancer, colon cancer, breast cancer, fallopian tube carcinoma, uterus Endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic Or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, pituitary adenoma, or these cancers A pharmaceutical composition comprising one or more combinations of. A pharmaceutical composition for inflammatory diseases comprising the general formula compound according to any one of claims 1 to 7 as an active ingredient and a pharmaceutically acceptable carrier.