JPWO2019160127A1 - Uses of DNMT inhibitors - Google Patents

Abstract

PROBLEM TO BE SOLVED: To replace a hydrolyzable transferase that is clinically used as a therapeutic agent for high-risk myelodysplastic syndrome and acute myelodysplastic syndrome ("Vidaza (registered trademark)" and "Dacogen (registered trademark)"). ATL therapeutic agents or prophylaxis for compounds that are highly stable to citidine deaminase, are absorbed into the body even by oral administration, are incorporated into the nucleic acid biosynthesis route, and have the effect of inhibiting the DNA methyltransferase DNMT. To provide as a medicine. SOLUTION: A prophylactic / therapeutic agent for ATL containing a compound of the following formula (I) or a salt thereof is provided. (In the formula, R is an OR3 group or a hydrogen atom, R1, R2 and R3 are hydrogen atoms or formula (II), respectively: (In the formula, R4, R5 and R6 each have a substituent. It is also a good alkyl group or an aryl group or an arylalkyl group). However, R1, R2 and R3 are excluding cases where they are hydrogen atoms at the same time.) [Selection diagram] None.

Description

The present invention relates to novel uses of an orally administrable DNMT inhibitor that has high stability to the hydrolytic metabolizing enzyme cytidine deaminase and can replace 5-azacitidine and its 2'-deoxy form.

ATL (Adult T-cell leukemia / lymphoma) is a disease discovered and named by Takatsuki et al. (Non-Patent Documents 1 and 2) in 1976, and is a neoplastic virus HTLV-1. Leukemia or malignant lymphoma caused by infection with (Human T-cell Leukemia / Lymphotropic Virus type-1). In Japan, HTLV-1 infections are predominant in the southern part (especially Kyushu and Okinawa), and are also localized in Caribbean coastal countries, Central Africa and South America.
Breastfeeding, sexual intercourse, blood transfusion, etc. are listed as transmission routes for HTLV-1, and it is said that there are 5 to 10 million carriers worldwide (about 1.1 million in Japan), and among male carriers in Japan. 6-7% of women and 2-3% of women develop ATL every year. The period from HTLV-1 infection to the onset of ATL is very long, and as a result, many elderly carriers develop ATL (Non-Patent Document 3).

Chemotherapy such as CHOP therapy (Non-Patent Document 4), modified LSG15 therapy (Non-Patent Document 5), EPOCH therapy (Non-Patent Document 6) and AZT-INFα therapy is selected for the treatment of ATL, but recurrence and drug resistance The current situation is that standard treatment methods have not yet been established.
Recently, a new ATL therapeutic drug, a chemokine receptor CCR4 antibody drug, mogamulizumab (product name: Poterigio) (Non-Patent Document 7), an antiviral drug, abacavir (Non-Patent Document 8), histone deacetylase (HDAC) inhibitor Vorinostat (product name: Zolinza) (Non-Patent Document 9) and the double inhibitor of histone deacetylase EZH1 / 2, DS-3201b, are attracting attention.

DNMTs is an abbreviation for DNA methyltransferases, methylation of the 6-position amino group of the adenine ring in the DNA strand (Adenine N ⁶ -specific DNA-methyltransferase: EC 2.1.1.72), cytosine. Catalyzes methylation of the ring 4-position amino group (Cytosine N ^4- specific DNA-methyltransferase: EC 2.1.1.113) or methylation to the cytosine ring 5-position (Cytosine C ^5- specific DNA-methyltransferase: EC 2.1.1.37) It is a group of enzymes. In particular, a group of enzymes that catalyze methylation to the 5-position of the cytosine ring in the sequence portion called CpG island, which is often found in the promoter region of the expressed gene (maintenance methyltransferase DNMT1 and de novo methyltransferase DNMT3 family). Plays a crucial role in regulating the normal development and differentiation of cells (Non-Patent Documents 10-11).

DNMTs are also deeply involved in cancer development. That is, it is thought that 60 to 90% of all CpGs have methylation at the 5-position of the cytosine ring, but abnormal levels of DNA methylation are closely related to the silencing of expressed genes, and the promoter region. It has been clarified that the transcription and expression of a gene in which (CpG island) is methylated at the 5-position of the cytosine ring at a high level is silenced (Non-Patent Documents 12 to 14).

On the other hand, cells are equipped with a mechanism to introduce a methyl group to the 5-position of the cytosine ring at the same position in the newly created DNA strand, and it is also DNMTs that enable this "replication of DNA methylation". .. Therefore, in cancerous cells, most of the tumor suppressor genes are suppressed in transcription and expression, and are in a silencing state, so that they can easily proliferate.

Regarding the methylation of the cytosine ring 5 position, the SH group of the cysteine residue at the catalytically active center of DNMT attacks the cytosine ring 6 position in the DNA sequence, thereby activating the cytosine ring 5 position and methylating. A reaction mechanism has been proposed that promotes methyl group transfer from the group donor S-adenosyl-L-methionine.

As selective enzyme inhibitors for DNMTs with such a background, 5-azacitidine (product name: "Vidaza (registered trademark)") and its 2'-deoxy compound (decitabine, product name: "Dacogen (registered trademark)") ) ”) Has been found and is clinically used as a therapeutic agent for high-risk myelodysplastic syndrome and acute myeloid leukemia. These drugs are very similar in chemical structure to cytosine nucleosides (the structure in which the carbon atom at the 5-position of the cytosine ring is replaced with a nitrogen atom), and DNA is used instead of 2'-deoxycytidine via the nucleic acid biosynthesis route. By entering the inside, it self-inhibits the methylation reaction at the 5-position of the cytosine ring by DNMTs in the tumor suppressor gene promoter region (CpG island), enabling normal expression of the tumor suppressor gene and showing a therapeutic effect. It is said to be forgotten.

Drugs with such a mechanism of action can be used as a wide range of anticancer agents, but all compounds are easily hydrolyzed by the metabolic enzyme citidine deaminase present in blood and liver. Due to its drawback of being deaminated, it remains for clinical use as a high-risk therapeutic agent for myelodysplastic syndrome and acute myelodysplastic syndrome, and because of its chemical instability, it is a dosage form as an injection. The current situation is that it remains at. Therefore, the emergence of an orally administrable drug that has high stability to cytidine deaminase and can replace 5-azacitidine and its 2'-deoxy form is desired.

Recently, SGI-110 (guadecitabin) (Patent Documents 1 and 2) has been found as a compound having high stability to the hydrolytic metabolizing enzyme cytidine deaminase, and a pro of 5-aza-2'-deoxycytidine Although clinical development is underway as a drug, this compound is extremely polar due to its dinucleotide structure, is not easily permeated through the membrane, and is not suitable as an oral administration agent (Non-Patent Documents 15 to 16). ..

Published in the United States 2007072996 (Japanese Patent No. 5030958) International Publication No. 2013303176 (Japanese Patent No. 6038921)

International Medicine, 1995, Vol. 34, No. 10, p. 947-952. Blood, 2017, Vol. 129, No. 9, p. 1071-1081. Cancer Science, 2017, Vol. 108, No. 9, p. 1719-1725. International Journal of Hematology, 2016, Vol. 104, No. 4, p. 468-475. Leukemia Research and Treatment, 2012, Paper, Registration No. 101754. Rinsho Ketsueki, 1998, Vol. 39, No. 4, p. 267-272. Cancer Science, 2017, Vol. 108, No. 10, p. 2022-2029. Scientific Reports, 2017, Vol. 7, No. 1, p. 12849. International Journal of Molecular Sciences, 2017, Vol. 18, No. 7, p. 1414. Chemical Reviews, 2015, Vol. 115, No. 6, p. 2240-2254. Biomolecules, 2017, Vol. 7, No. 1, p. 3. 3. Book "Epigenetics: Reference Manual" (Epigenetics: A Reference Manual), 2011, by Craig JM, Wang NC, Kaister Academic Press. Cell & Bioscience, 2014, Volume 4, p. 46. Molecular Cancer, 2017, Vol. 16, p. 29. Oncotarget, 2017, Vol. 8, No. 2, p. 2949-2959. Epigenetics, 2016, Vol. 11, No. 10, p. 709-720.

The subject of the present invention is to create an orally administrable compound which has high stability to the hydrolyzable metabolizing enzyme cytidine deaminase and can replace 5-azacitidine and its 2'-deoxy form, and has a high risk. It is intended to be provided not only as a therapeutic agent for myelodysplastic syndrome and acute myeloid leukemia, but also as a new therapeutic agent or a prophylactic agent for ATL.
The onset of ATL plays an important role in the intercellular adhesion molecule ICAM-1 (Int. J. Cancer, 1995, 60 (4), 554-561.), Tumor progression, and immunomodulation and signal transduction. CD26 / DPPIV related to apoptosis (Int. J. Hematol., 2004, 80 (3), 254-260.), DNA mismatch repair protein MLH1 (Oncol. Rep., 2005, 14 (1), 191-194. ), Bone-forming protein BMP-6 (Int. J. Cancer, 2008, 123 (8), 1824-1831.) And nuclear transduction factor PDLIM2 (Neoplasia, 2009, 11 (10), 1036-) essential for inflammatory response. 1041.) Molecules, or 5'LTR (J. Virol., 2002, 76 (18), 9389-9397.) Used by viruses to insert their own genetic material into the host genome, and normal cells Reported that hypermethylation of genes such as p16 [INK4a] (Int. J. Mol. Med., 2011, 28 (5), 835-839.) Related to self-defense mechanism to prevent canceration is involved. However, there is no example in which a DNMT inhibitor related to the present invention has been positively investigated as a novel ATL treatment or preventive agent.

The present inventors include 5-azacitidine (product name: "Vidaza (registered trademark)") and its 2'-deoxy compound (product name:) as therapeutic agents for various bone marrow tumors including high-risk myelodysplastic syndrome. Since it provides a drug that is more useful than "Dacogen®"), it has high stability to the hydrolyzable metabolizing enzyme citidine deaminase and is excellent in being able to easily enter the nucleic acid biosynthesis route in vivo. We have been diligently researching to find new compounds that have both pharmacological action and physicochemical properties. Among them, various sugar-modified derivatives of 5-azacitidine and its 2'-deoxy compound were synthesized, and their chemical reactivity and biological activity were investigated. As a result, the corresponding sugar-substituted silyl ether derivative was citidine. We have found that it has excellent stability to deaminase (see Patent No. 6162349) and exhibits anti-ATL activity, and based on these findings, further detailed studies have been carried out to complete the present invention. It was.

That is, when 5-azacitidine and various sugar-modified derivatives of its 2'-deoxy compound were evaluated using a recently developed simple anti-ATL activity screening evaluation system, 5-aza-2'-deoxycytidine was evaluated. It was confirmed that all of the sugar moiety silyl ether derivatives of AZ had very high anti-ATL activity. This anti-ATL activity was interpreted as the result of these compounds being non-enzymatically activated in the target ATL cells and incorporated into DNA via the nucleic acid biosynthetic route. In addition, since these 5-aza-2'-deoxycytidine sugar moiety silyl ether derivatives are highly lipophilic and have physicochemical properties that can be orally administered, all of these compounds have DNMT. It can be an orally administrable therapeutic agent for the associated ATL. In addition, from the consideration of its mechanism of action (restoration of tumor suppressor genes by DNMT inhibition), it was speculated that it could be used as a preventive drug for ATL associated with DNMT.
Based on these findings, the present invention solves the above problems by providing the inventions described below.
[1]
Equation (I):


(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:


(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ATL prophylactic / therapeutic agent containing the compound represented by) or a salt thereof.
[2]
Prevention of ATL containing the compound according to [1] or a salt thereof, wherein R ¹ is a silyl group represented by the formula (II) and R ² and R or R ^{3 are hydrogen atoms.} Therapeutic agent.
[3]
Prevention of ATL containing the compound according to [1] or a salt thereof, wherein R ² is a silyl group represented by the formula (II) and R ¹ and R or R ^{3 are hydrogen atoms.} Therapeutic agent.
[4]
Prevention of ATL containing the compound according to [1] or a salt thereof, wherein R ¹ and R ² are silyl groups represented by the formula (II), respectively, and R or R ^{3 is a hydrogen atom.}・ Therapeutic agent.
[5]
A preventive / therapeutic agent for ATL containing the compound according to [1] or a salt thereof, wherein R ¹ is a hydrogen atom and R ² and R ^{3 are silyl groups represented by the formula (II), respectively.}
[6]
A preventive / therapeutic agent for ATL containing the compound according to [1] or a salt thereof, wherein R ¹ , R ² and R ^{3 are silyl groups represented by the formula (II), respectively.}
[7]
The R ⁴ , R ⁵ and R ⁶ _{are C 1 to} C ₈ alkyl groups or C _{6 to} C ₁₀ aryl groups or C _{7 to} C ₁₄ aryl alkyl groups, respectively, which may have a substituent [1]. ] ATL prophylactic / therapeutic agent.
[8]
The prophylactic / therapeutic agent for ATL according to [7], wherein the C _{6 to} C _{10 aryl groups are phenyl groups or naphthyl groups.}
[9]
The _C 7 _{-C 14} arylalkyl group, a benzyl group, a phenethyl group or a naphthylmethyl group, an agent for the prophylaxis or treatment of ATL described in [7].
[10]
Equation (I):


(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ), A method for preventing or treating ATL, which comprises administering an effective amount of a compound or a salt thereof.
[11]
Formula (I) for producing a pharmaceutical composition for the prevention or treatment of ATL:

(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ) Or a salt thereof.

According to the present invention, 5-azacitidine or its 2'-deoxysaccharide silyl ether derivative is more lipophilic than the corresponding 5-azacitidine or its 2'-deoxy, and can be administered orally. After being absorbed in the intestine, it is non-enzymatically hydrolyzed and activated in the cell membrane or intracellular of ATL cells without being affected by the hydrolytic metabolic enzyme citidine deaminase in the blood or liver. Since it is presumed that it exhibits DNMT inhibitory activity by being incorporated into DNA via the nucleic acid biosynthesis route, it can be expected to function as a therapeutic or prophylactic agent for ATL whose expression is induced by DNMT.

Unless otherwise specified, the terms used herein and in the claims have the following meanings.

The compound of the present invention or a salt thereof The compound of the present invention is a compound represented by the following formula (I).


(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ) Is a compound or a salt thereof.

Unless otherwise specified, the "alkyl group" refers to a saturated aliphatic hydrocarbon group, for example, a linear or branched or cyclic alkyl group having 1 to 8 carbon atoms, for example, a methyl group or an ethyl group. , propyl group, isopropyl group, butyl group, sec- butyl group, an isobutyl group, tert- butyl group, a pentyl group, _C 1 -C ₆ alkyl groups such as hexyl, heptyl, 2-methylhexyl group, 5-methyl Hexyl group, 2,2-dimethylpentyl group, 4,4-dimethylpentyl group, 2-ethylpentyl group, 1,1,3-trimethylbutyl group, 1,2,2-trimethylbutyl group, 1,3,3 -Trimethylbutyl group, 2,2,3-trimethylbutyl group, 2,3,3-trimethylbutyl group, 1-propylbutyl group, 1,1,2,2-tetramethylpropyl group, octyl group, 2-methyl Heptyl group, 3-methylheptyl group, 6-methylheptyl group, 2-ethylhexyl group, 5,5-dimethylhexyl group, 2,4,4-trimethylpentyl group, 1-ethyl-1-methylpentyl group, cyclopropyl , Cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like can be mentioned, but groups of C _{1 to} C ₆ alkyl are preferable. Preferred examples of C _{1 to} C ₆ alkyl groups are methyl, ethyl and propyl groups. Further, preferred examples of the cyclic alkyl group are a cyclopentyl group and a cyclohexyl group.

The "aryl" refers to a monocyclic or bicyclic aromatic hydrocarbon, preferably a C _6-10 aryl group such as a phenyl group or a naphthyl group, and more preferably a phenyl group.

"Arylalkyl" means an alkyl group substituted with aryl. _Preferably, a C 7 _{-C 14} arylalkyl group. Examples of C _{7 to} C ₁₄ arylalkyl groups include, but are not limited to, benzyl group, phenethyl group, naphthylmethyl group and the like.

The "alkyl group which may have a substituent, the aryl group which may have a substituent or the arylalkyl group which may have a substituent" means that even if it has a substituent, It may be unsubstituted. When substituted, the substituents may have 1 to 5, preferably 1-3, at substitutable positions of the alkyl group, aryl group or arylalkyl group, and the number of substituents is 2 or more. In some cases, each substituent may be the same or different. Examples of the substituent include an alkyl group, a halogen atom, a cyano group, a nitro group and the like, and an example of a preferable substituent is an alkyl group or a halogen.

The “halogen atom” refers to a fluorine atom, a chlorine atom, a bromine atom, an iodine atom and the like, and preferred examples are a fluorine atom and a chlorine atom.

The salt of the compound represented by the formula (I) of the present invention may be any salt as long as it is pharmacologically acceptable. Examples of the salt include inorganic acid salts (for example, hydrochlorides, sulfates, hydrobromates, phosphates, etc.), organic acid salts (for example, acetates, trifluoroacetates, succinates, etc.). Acid addition salts such as maleate, fumarate, propionate, citrate, tartrate, lactate, oxalate, methanesulfonate, p-toluenesulfonate, etc.) can be mentioned. It is not limited to.

The compound represented by the formula (I) of the present invention may be a crystal, a single crystal form, or a mixture of a plurality of crystal forms. Crystals can be produced by crystallization by applying a crystallization method known per se.

Further, the compound represented by the formula (I) of the present invention may be a solvate (for example, a hydrate or the like), and may be a solvate or a non-solvate (for example, a non-hydrate or the like). Is also included in the compound represented by the formula (I).

The 5-azacitidine and its 2'-deoxy sugar silyl ether derivative of the present invention (see formula (I)) can be prodrugs of 5-azacitidine and its 2'-deoxy.

Since 5-azacitidine and its 2'-deoxy sugar sugar silyl ether derivatives (see formula (I)) are very stable to cytidine deaminase, these derivatives absorbed from the gastrointestinal tract It is expected to have the property of being less susceptible to hydrolytic metabolism by the enzyme cytidine deaminase in the blood and liver.

High stability to the above hydrolyzable metabolizing enzymes is expected, and 5-azacitidine and its 2'-deoxy sugar moiety silyl ether derivative (see formula (I)) according to the present invention are expressed by DNMT. It can be a therapeutic or prophylactic agent for the induced ATL.

Method for producing compound represented by formula (I) of the present invention

The 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) according to the present invention can be produced by the following method. That is, 5-azacitidines (see formula (I): R = OR ³ or H, R ¹ = R ² = R ³ = H) and a silyl chloride having an appropriate substituent in the presence of a dehydrohalogenating agent. By reacting with, the desired 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) can be easily obtained.

(Dehalogenating agent) Examples of the dehydrohalizing agent to be used include organic bases and inorganic bases, and the organic bases are not limited to these, but imidazole, 1-methylimidazole, triethylamine, N, N. -Diisopropylethylamine, pyridine, 4-dimethylaminopyridine (DMAP), n-butyllithium or potassium-tert-butoxide, etc., and examples of the inorganic base include, but are not limited to, sodium hydride, sodium carbonate, carbonate. Examples thereof include sodium hydride, potassium carbonate, potassium hydrogen carbonate, cesium carbonate and the like. The amount of the base used is preferably equal to or more than the equivalent amount of the raw material compound. Further, the range of 1.0 to 50.0 equivalents can be exemplified with respect to 1 mol of the raw material compound, but the range of 1.0 to 10.0 equivalents is preferable, and more preferably 1.0 to 5. It is preferably in the range of 0 equivalents.

(solvent)
From the viewpoint of smooth progress of the reaction, the reaction of the present invention is preferably carried out in the presence of a solvent. The solvent in the reaction of the present invention may be any solvent as long as the reaction proceeds.
Examples of the solvent include N, N-dimethylformamide, N, N-dimethylacetamide and dimethyl sulfoxide. The amount of the solvent used may be any amount as long as the reaction proceeds. The amount of solvent used in the reaction of the present invention can be appropriately adjusted by those skilled in the art.

(Reaction temperature)
The reaction temperature of the present invention is not particularly limited. In one embodiment, from the viewpoint of improving yield, suppressing by-products, economic efficiency, etc., -20 ° C to 50 ° C (that is, -20 ° C to + 50 ° C), preferably -10 ° C to 30 ° C. (Ie, minus 10 ° C to plus 30 ° C), more preferably -10 ° C to 20 ° C (ie, minus 10 ° C to plus 20 ° C), even more preferably -5 ° C to 15 ° C (ie, minus 5 ° C to plus). 15 ° C.), particularly preferably in the range of −5 ° C. to 10 ° C. (that is, −5 ° C. to −10 ° C.).

(Reaction time)
The reaction time of the present invention is not particularly limited. In one embodiment, from the viewpoint of improving the yield, suppressing by-products, economic efficiency, etc., 0.5 hour to 120 hours, preferably 1 hour to 72 hours, more preferably 1 hour to 48 hours, and further. Preferably, the range of 1 hour to 24 hours can be exemplified. However, the reaction time of the present invention can be appropriately adjusted by those skilled in the art.

Pharmaceutical Composition of the Present Invention The compound represented by the formula (I) of the present invention can be used as it is or mixed with a pharmacologically acceptable carrier by a method known per se to prepare a pharmaceutical composition for mammals (for example, a mammal (for example). , Humans, monkeys, cats, pigs, horses, cows, mice, rats, guinea pigs, dogs, rabbits, etc.) can be used as a safe drug.

Here, as the pharmacologically acceptable carrier, various conventional organic or inorganic carrier substances are used as the preparation material, and examples thereof include excipients, lubricants, binders and disintegrants in solid preparations. Examples thereof include solvents, solubilizing agents, suspending agents and the like, tensioning agents and buffering agents in liquid preparations. Further, if necessary, preparation additives such as preservatives, antioxidants, colorants and sweeteners can also be used.

Examples of the dosage form of the pharmaceutical composition include tablets, capsules (including soft capsules and microcapsules), granules, powders, syrups, emulsions, suspensions, oral preparations such as sustained-release agents, and the like. , These can be safely administered orally. However, this does not apply because liquid preparation is also possible.

The pharmaceutical composition can be produced by a method commonly used in the field of formulation technology, for example, the method described in the Japanese Pharmacopoeia.

Uses of the compound represented by the formula (I) of the present invention The compound represented by the formula (I) of the present invention has many therapeutic and prophylactic uses. In a preferred embodiment, the compounds of the invention are used in the treatment of a wide variety of diseases that are sensitive to treatment with cytidines (eg, 5-azacitidine and 5-aza-2'-deoxycytidine). Preferred indications that can be treated with the compounds of the invention include those with unwanted or uncontrolled cell division. Such indications include a variety of cancers, but more preferably ATLs whose expression is evoked by DNMTs.

Suitable pharmaceutical compositions used in the present invention include compositions in which the active ingredient is present in an effective amount, i.e., in an amount effective to achieve therapeutic and / or prophylactic objectives in the condition being treated. Is included.

The pharmaceutical composition used in the present invention is provided as a dosage form for oral administration. The pharmaceutical compositions provided herein may be provided in solid, semi-solid or liquid dosage form for oral administration. As used herein, oral administration also includes buccal, tongue and sublingual administration. Suitable oral dosage forms include tablets, capsules, pills, troches, medicated candy, aromatics, cashiers, pellets, drug-added chewing gums, granules, powders, effervescent or non-foaming powders or granules. , Solutions, emulsions, suspensions, solutions, wafers, sprinkles, elixirs and syrups, but not limited to these. In addition to the active ingredient, the pharmaceutical composition comprises binders, fillers, diluents, disintegrants, wetting agents, lubricants, flow promoters, colorants, pigment migration inhibitors, sweeteners and flavoring agents. It may contain a carrier or excipient that is acceptable as one or more pharmaceuticals, not limited to these.

The amount of the compound represented by the formula (I) of the present invention in the pharmaceutical composition or dosage form is, for example, about 1 mg to about 2,000 mg, about 10 mg to about 2,000 mg, about 20 mg to about 2,000 mg, about 50 mg to about 1,000. It may be in the range of mg, about 100 mg to about 500 mg, about 150 mg to about 500 mg or about 150 mg to about 250 mg.
When the compound of the present invention is used as an anticancer agent, the effective dose thereof is the nature of the cancer, the degree of cancer progression, the treatment policy, the degree of metastasis, the amount of tumor, the body weight, the age, the sex and the patient's ( Although it can be appropriately selected depending on the genetic) racial background and the like, the pharmaceutically effective amount is generally determined based on factors such as clinically observed symptoms and the degree of cancer metastasis. The daily dose, for example, when administered to humans, is about 0.01 mg / kg to about 10 mg / kg (for an adult weighing 60 kg, about 0.5 mg to about 500 mg), preferably about 0.05 mg / kg to. It is about 5 mg / kg, more preferably about 0.1 mg / kg to about 2 mg / kg. The administration may be performed once or in multiple doses.
When the stability of the 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) thus obtained in the presence of citidine deaminase was examined, it has a sugar moiety silyl ether group according to the present invention. In all cases, the derivatives were found to be very stable in the presence of cytidine deaminase, and these 5-azacitidine sugar silyl ether derivatives are less susceptible to hydrolytic metabolism by the enzyme cytidine deaminase in blood and liver. I was able to confirm that. On the other hand, 5-azacitidine having a hydroxyl group at the 5'position and its 2'-deoxy compound (see formula (I): R = OR ³ or H, R ¹ = R ² = R ³ = H) were used. Under the conditions, it decomposed within 30 minutes.
Further, the stability of the 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) thus obtained was examined in an environment close to physiological conditions (for example, 37 ° C. in PBS solution). As a result, if a substituent directly linked to a silyl group is successfully selected from the derivatives according to the present invention, it is hydrolyzed at an appropriate speed, and the corresponding 5-azacitidines (see formula (I): R = OR ³ or It was confirmed that H, R ¹ = R ² = R ³ = H) was efficiently generated. It has also been confirmed that the 5-azacitidine sugar moiety silyl ether derivative, which is hydrolyzed at an appropriate speed, exhibits anti-ATL activity.

Therefore, the 5-azacitidine sugar moiety silyl ether derivative according to the present invention, which has high stability to the above-mentioned hydrolyzable metabolic enzyme and has appropriate hydrolysis reactivity under physiological conditions (formula (I). ) Can be an ATL therapeutic or prophylactic agent.

Details of experiments on the production of these 5-azacitidine sugar silyl ether derivatives (see formula (I)), stability to the metabolic enzyme cytidine deaminase, and hydrolysis reactivity in PBS solution, and anti-ATL activity are shown below. ..
Example

Hereinafter, the present invention will be described in more detail with reference to examples, but these are not intended to limit the present invention.

In the following examples, room temperature means about 15-30 ° C. ^{1 1} H-NMR and ¹³ C-NMR spectra were measured using JEOL JNM-ECZ 400R and chemically shifted from the internal standard tetramethylsilane using _{CDCl 3} , DMSO-d ₆ or CD _{3 OD as a solvent.} It showed δ (ppm). Other symbols in the present specification have the following meanings. s: Singlet d: Doublet t: Triplet m: Multiplet br: Broad br s: Broad singlet J: Coupling constant In addition, the Mass spectrum data of each compound was measured using a Yamazen Smart Flash MS system device (APCI method). The value. The reaction time, column elution solvent system, isolation yield and instrument data for the 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) obtained in this study are shown below.

Synthesis of 3', 5'-disilyloxy-5-aza-2'-deoxycytidines


Add imidazole (2 mM) to a suspension of 5-aza-2'-deoxycytidine (see formula (I): R ¹ = R ² = R = H) (1 mM) in anhydrous N, N-dimethylformamide (3 mL). After the addition, the corresponding silyl chloride (2.5 mM) is added dropwise under ice-cooling over about 10 minutes, and then the mixture is gradually returned to room temperature and stirred until the raw materials disappear. The reaction solution is poured into 50 mL of a mixed solution of ethyl acetate-saturated saline (2: 1) and extracted with ethyl acetate. The extract was washed with saturated brine (10 mL each, twice), dried over anhydrous sodium sulfate, and the insoluble material was filtered off. The extract was dried under reduced pressure to obtain an oily residue on silica gel. By separating and purifying with a pack column (Yamazen Smart Flash MS system device), the target 5-aza-2'-deoxycitidine 3', 5'-disilyl ether derivative (in the formula (I), R is A compound which is a hydrogen atom and R ¹ and R ² are silyl groups.) Can be isolated as a white powder.
3', 5'-di (O-trimethylsilyl) -5-aza-2'-deoxycytidine: 3', 5'-Di (O-trimethylsilyl) -5-aza-2'-deoxycytidine: (Equation (I)) Medium, R = H, R ¹ = R ² = Trimethylsilyl group) (Reaction time: about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 70%)
^{1 1} H-NMR (CDCl ₃ ): 8.69 (s, 1H), 6.17 (dd, J = 6.4 and 4.4Hz, 1H), 5.89 (br s, 1H), 5.44 (br s, 1H), 4.36 (q, J = 5.6Hz, 1H), 3.94-3.96 (m, 1H), 3.88 (dd, J = 11.6 and 2.8Hz, 1H), 3.71 (dd, J = 12.0 and 2.4Hz), 2.50 (q, J = 6.8) Hz, 1H), 2.17-2.23 (m, 1H), 0.16 (s, 9H), and 0.12 (s, 9H).
¹³ C-NMR (CDCl ₃ ): 166.4, 156.2, 154.0, 87.6, 86.6, 69.7, 60.8, 42.2, 0.10, and -0.69.
Mass: 373.3 [M + H] ⁺ (Calcd. For C ₁₄ H ₂₈ N ₄ O ₄ Si ₂ , MW = 372.16).
3', 5'-di (O-triethylsilyl) -5-aza-2'-deoxycytidine: 3', 5'-Di (O-triethylsilyl) -5-aza-2'-deoxycytidine: (Formula (I) ), R = H, R ¹ = R ² = Triethylsilyl group) (Reaction time: about 2 hours, column elution solvent: ethyl acetate-n-hexane system, isolation yield: 54%)
¹ H-NMR (CDCl ₃ ): 8.67 (s, 1H), 6.19 (dd, J = 6.4 and 4.8Hz, 1H), 5.61 (br, 1H), 5.38 (br, 1H), 4.41 (q, J = 4.8Hz, 1H), 3.96-3.98 (m, 1H), 3.91 (dd, J = 11.6 and 2.8Hz, 1H), 3.76 (dd, J = 11.6 and 2.0Hz, 1H), 2.51 (dt, J = 13.2) and 6.0Hz, 1H), 2.15-2.21 (m, 1H), 0.92-0.99 (m, 18H), and 0.56-0.68 (m, 12H).
¹³ C-NMR (CDCl ₃ ): 166.4, 156.2, 154.0, 88.0, 86.6, 70.2, 61.5, 42.7, 6.8, 4.7, and 4.2.
Mass: 457.4 [M + H] ⁺ (Calcd. For C ₂₀ H ₄₀ N ₄ O ₄ Si ₂ , MW = 456.26).
3', 5'-Di (O-normal octyldimethylsilyl) -5-aza-2'-deoxycytidine: 3', 5'-Di (On-octyldimethylsilyl) -5-aza-2'-deoxycytidine: (Formula In (I), R = H, R ¹ = R ² = n-Octyldimethylsilyl group) (reaction time: about 2 hours, column elution solvent: ethyl acetate-n-hexane system, isolation yield: 54%)
^{1 1} H-NMR (CD ₃ OD): 8.61 (s, 1H), 6.10 (t, J = 5.2Hz, 1H), 4.46 (dd, J = 10.0 and 4.8Hz, 1H), 3.97 (dd, J = 6.4) and 2.8Hz, 1H), 3.88 (dd, J = 11.6 and 3.2Hz. 1H), 3.76 (dd, J = 11.2 and 2.4Hz, 1H), 2.41 (dt, J = 13.6 and 6.0Hz, 1H), 2.24 (dt, J = 13.6 and 5.6Hz, 1H), 1.29-1.34 (m, 24H), 0.87-0.91 (m, 6H), 0.61-0.68 (m, 4H), 0.14 (s, 6H), and 0.12 ( s, 6H).
¹³ C-NMR (CD ₃ OD): 166.7, 155.8, 155.0, 88.0, 86.5, 70.8, 61.2, 41.6, 33.3, 31.8, 29.16, 29.12, 29.11, 23.0, 22.9, 22.4, 16.0, 15.6, 13.2, -2.78 , -2.89, -3.57, and -3.75.
Mass: 569.5 [M + H] ⁺ (Calcd. For C ₂₈ H ₅₆ N ₄ O ₄ Si ₂ , MW = 568.38).

Synthesis of 2', 3', 5'-trisilyloxy-5-azacitidines


5-Azacitidine (see formula (I): R = OR ³ , R ¹ = R ² = R ³ = H) (1 mM) in anhydrous N, N-dimethylformamide (2 mL) suspension with imidazole (4 mM). After the addition, the corresponding silyl chloride (3.5 mM) is added dropwise under ice-cooling over about 10 minutes, then the mixture is gradually returned to room temperature and stirred until the raw materials disappear. The reaction solution is poured into 50 mL of a mixed solution of ethyl acetate-saturated saline (2: 1) and extracted with ethyl acetate. The extract was washed with saturated brine (10 mL each, twice), dried over anhydrous sodium sulfate, and the insoluble material was filtered off. The extract was dried under reduced pressure to obtain an oily residue on silica gel. By separating and purifying with a pack column (Yamazen Smart Flash MS system device), the target 5-azacitidine 2', 3', 5'-trisilyl ether derivative (in formula (I), R is ^{3 ORs).} , and the compound R ^1, R ² and R ³ is a silyl group.) it can be isolated as a white powder.
2', 3', 5'-tri (O-trimethylsilyl) -5-azacytidine: 2', 3', 5'-Tri (O-trimethylsilyl) -5-azacytidine: (in formula (I), R ¹ = R ² = R ³ = Trimethylsilyl group) (Reaction time: about 1 hour, column elution solvent: ethyl acetate-n-hexane system, isolation yield: 64%)
^{1 1} H-NMR (CDCl ₃ ): 8.82 (s, 1H), 6.23 (br, 1H), 5.70 (s, 1H), 5.49 (br, 1H), 4.09-4.16 (m, 3H), 4.01 (dd, dd, J = 12.0 and 1.2Hz, 1H), 3.70 (dd, J = 11.6 and 1.2Hz, 1H), 0.20 (s, 9H), 0.19 (s, 9H), and 0.13 (s, 9H).
¹³ C-NMR (CDCl ₃ ): 166.5, 156.4, 153.9, 91.2, 82.7, 76.4, 68.3, 59.3, 0.4, 0.2, and -0.7.
Mass: 461.3 [M + H] ⁺ (Calcd. For C ₁₇ H ₃₆ N ₄ O ₅ Si ₃ , MW 460.20).
2', 3', 5'-tri (O-ethyldimethylsilyl) -5-azacitidine: 2', 3', 5'-Tri (O-ethyldimethylsilyl) -5-azacytidine: (R in formula (I)) ¹ = R ² = R ³ = Ethyldimethylsilyl group) (Reaction time: about 1 hour, column elution solvent: ethyl acetate-n-hexane system, isolation yield: 67%)
¹ H-NMR (CDCl ₃ ): 8.80 (s, 1H), 6.27 (br, 1H), 5.71 (d, J = 0.8Hz, 1H), 5.49 (br, 1H), 4.08-4.16 (m, 3H) , 4.01 (dd, J = 12.0 and 0.8Hz, 1H), 3.72 (dd, J = 11.6 and 0.8Hz, 1H), 0.90-1.01 (m, 9H), 0.57-0.74 (m, 6H), 0.19 (s) , 3H), 0.16 (s, 9H), 0.10 (s, 3H), and 0.09 (s, 3H).
¹³ C-NMR (CDCl ₃ ): 166.5, 156.3, 153.9, 91.1, 82.8, 68.4, 59.6, 8.6, 8.3, 7.7, 6.8, -1.8, -1.9, -2.1, -2.8, and -3.0.
Mass: 503.4 [M + H] ⁺ (Calcd. For C ₂₀ H ₄₂ N ₄ O ₅ Si ₃ , MW = 502.25).
2', 3', 5'-tri (O-isopropyldimethylsilyl) -5-azacitidine: 2', 3', 5'-Tri (Oi-propyldimethylsilyl) -5-azacytidine: (in formula (I), R ¹ = R ² = R ³ = i-Propyldimethylsilyl group) (Reaction time: Approximately 1 hour, Column elution solvent: Ethyl acetate-n-hexane system, Isolation yield: 74%)
¹ H-NMR (CDCl ₃ ): 8.76 (s, 1H), 6.68 (br, 1H), 5.71 (d, J = 1.2Hz, 1H), 5.55 (br, 1H), 4.09-4.17 (m, 3H) , 4.03 (d, J = 12.0Hz, 1H), 3,74 (d, J = 11.6Hz, 1H), 0.92-1.02 (m, 21H), 0.18 (s, 3H), 0.14 (s, 3H), 0.12 (s, 3H), 0.11 (s, 3H), and 0.07 (s, 6H).
¹³ C-NMR (CDCl ₃ ): 166.5, 156.2, 153.9, 90.9, 83.0, 76.4, 68.7, 59.9, 17.0, 16.9, 14.9, 14.6, 14.3, -3.4, -3.5, -3.9, -4.1, -4.5, and -4.8.
Mass: 545.4 [M + H] ⁺ (Calcd. For C ₂₃ H ₄₈ N ₄ O ₅ Si ₃ , MW = 544.29).
2', 3', 5'-Tri (O-tert-butyldimethylsilyl) -5-azacitidine: 2', 3', 5'-Tri (Ot-butyldimethylsilyl) -5-azacytidine: (in formula (I)) , R ¹ = R ² = R ³ = t-Butyldimethylsilyl group) (Reaction time: Approximately 15 hours, Column elution solvent: Ethyl acetate-n-hexane system, Isolation yield: 67%)
¹ H-NMR (CDCl ₃ ): 8.73 (s, 1H), 6.46 (br, 1H), 5.73 (d, J = 2Hz, 1H), 5.45 (br, 1H), 4.17 (dd, J = 3.6 and 1.6) Hz, 1H), 4.06-4.13 (m, 3H), 3.80 (d, J = 1.2Hz, 0.5H), 3.77 (d, J = 1.6Hz, 0.5H), 0.96 (s, 9H), 0.91 (s , 9H), 0.89 (s, 9H), 0.21 (s, 3H), 0.15 (s, 3H), 0.13 (s, 3H), 0.11 (s, 3H), and 0.06 (s, 3H).
¹³ C-NMR (CDCl ₃ ): 171.9, 161.7, 159.4, 95.9, 88.7, 81.5, 74.6, 66.3, 31.7, 31.4, 24.2, 23.6, 23.5, 1.45, 1.31, 0.52, 0.44, and 0.22.
Mass: 587.5 [M + H] ⁺ (Calcd. For C ₂₆ H ₅₄ N ₄ O ₅ Si ₃ MW = 586.34).
2', 3', 5'-tri (O-triethylsilyl) -5-azacitidine: 2', 3', 5'-Tri (O-triethylsilyl) -5-azacytidine: (in formula (I), R ¹ = R ² = R ³ = Triethylsilyl group) (Reaction time: Approximately 1 hour, Column elution solvent: Ethyl acetate-n-hexane system, Isolation yield: 99%)
¹ H-NMR (CDCl ₃ ): 8.78 (s, 1H), 5.87 (br, 1H), 5.73 (d, J = 1.2Hz, 1H), 4.10-4,17 (m, 3H), 4.04 (dd, dd, J = 11.6 and 1.6Hz, 1H), 3.77 (dd, J = 11.6 and 1.2Hz, 1H), 0.92-1.01 (m, 27H), and 0.57-0.78 (m, 18H).
¹³ C-NMR (CDCl ₃ ): 166.4, 156.3, 153.8, 90.6, 83.0, 76.4, 68.8, 60.2, 6.82, 6.80, 6.74, 4.80, 4.75, and 4.07.
Mass: 587.5 [M + H] ⁺ (Calcd. For C ₂₆ H ₅₄ N ₄ O ₅ Si ₃ , MW = 586.34).
2', 3', 5'-tri (O-isopropyldiethylsilyl) -5-azacitidine: (2', 3', 5'-Tri (Oi-propyldiethylsilyl) -5-azacytidine: (in formula (I), R ¹ = R ² = R ³ = i-Propyldiethylsilyl group) (Reaction time: Approximately 1 hour, Column elution solvent: Ethyl acetate-n-hexane system, Isolation yield: 74%)
¹ H-NMR (CDCl ₃ ): 8.76 (s, 1H), 6.38 (br, 1H), 5.75 (d, J = 2.0Hz, 1H), 5.47 (br, 1H), 4.07-4.22 (m, 4H) , 3.81 (d, J = 10.4Hz, 1H), 0.94-1.05 (m, 36H), and 0.63-0.76 (m, 15H).
¹³ C-NMR (CDCl ₃ ): 166.4, 156.4, 153.9, 90.3, 83.2, 69.3, 60.6, 17.4, 17.3, 13.1, 13.0, 12.4, 7.2, 7.1, 7.0, 3.9, 3.8, 3.7, 3.0, and 2.8.
Mass: 629.5 [M + H] ⁺ (Calcd. For C ₂₉ H ₆₀ N ₄ O ₅ Si ₃ , MW = 628.39).

Synthesis of silyloxy-5-azacitidines at the 5'position


5-Azacitidines (see formula (I): R = H or OR ³ , R ¹ = R ² = R ³ = H) (1 mM) in anhydrous N, N-dimethylformamide (3 mL) suspension with imidazole ( After adding (1.5 mM), the corresponding silyl chloride (1.2 mM) was added dropwise under ice cooling over about 10 minutes, then gradually returning to room temperature until the raw material disappeared (about 1 to 17 hours). ) Stir. The reaction solution is poured into 50 mL of a mixed solution of ethyl acetate-saturated saline (2: 1) and extracted with ethyl acetate. The extract was washed with saturated brine (10 mL each, twice), dried over anhydrous sodium sulfate, and the insoluble material was filtered off. The extract was dried under reduced pressure to obtain an oily residue of silica gel. By separating and purifying with a pack column (Yamazen Smart Flash MS system device), the 5'-position silyl ether derivative of the target 5-azacitidines (in formula (I), R is a hydroxyl group or a hydrogen atom, and R ¹ Is a silyl group and R ² is a hydrogen atom.) Can be isolated as a white powder.
Compound A: 5'-O- (Trimethylsilyl) -5-azacytidine (in formula (I), R ¹ = Trimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / isolation method: Reaction Time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 14%)
¹ H-NMR (CDCl ₃ ): 8.53 (s, 1H), 6.20 (br, 1H), 5.81 (d, J = 3.2Hz, 1H), 5.69 (br, 1H), 5.30 (br, 1H), 4.38 (s, 1H), 4.25 (s, 2H), 3.87 (d, J = 10.8Hz, 1H), 3.72 (d, J = 10.8Hz, 1H), 3.45 (br, 1H), and 0.09 (s, 9H) ).
¹³ C-NMR (CDCl ₃ ): 166.7, 155.9, 155.5, 93.3, 87.8, 78.1, 72.6, 62.1, and -0.82.
Mass: 317.2 [M + H] ⁺ (Calcd. For C ₁₁ H ₂₀ N ₄ O ₅ Si, MW = 316.12).
Compound B: 5'-O- (Trimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = Trimethylsilyl group, R = R ² = H) (Synthesis conditions / Isolation method: Reaction time = Approximately 1 hour, column elution solvent: ethyl acetate-methanol system)
^{1 1} H-NMR (CD ₃ OD): 8.66 (s, 1H), 6.13 (t, J = 6.0Hz, 1H), 4.35-4.42 (m, 1H), 3.67-4.02 (m, 9H), 2.34-2.50 (m, 1H), 2.20-2.32 (m, 1H), and 0.14 (s, 9H).
¹³ C-NMR (CDCl ₃ ): 166.3, 156.0, 154.1, 87.6, 86.8, 71.6, 62.3, 42.6, and 0.1.
Mass: 301.3 [M + H] ⁺ (Calcd. For C ₁₁ H ₂₀ N ₄ O ₄ Si, MW = 300.13).
Compound C: 5'-O- (Ethyldimethylsilyl) -5-azacytidine (in formula (I), R ¹ = Ethyldimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / isolation method: Reaction Time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 12%)
¹ H-NMR (CDCl ₃ ): 8.56 (s, 1H), 6.61 (br, 1H), 5.94 (br, 1H), 5.83 (d, J = 4.0Hz, 1H), 4.31-4.34 (m, 1H) , 4.23-4.28 (m, 2H), 3.91 (dd, J = 11.6 and 2.4Hz, 1H), 3.74 (dd, J = 11.6 and 2.4Hz, 1H), 0.92 (t, J = 8.0Hz, 3H), 0.56 (q, J = 8.0Hz, 2H), 0.09 (s, 3H), and 0.08 (s, 3H).
¹³ C-NMR (CDCl ₃ ): 166.5, 155.7, 155.6, 92.8, 87.3, 72.0, 62.0, 7.6, 6.6, and -3.03.
Mass: 331.2 [M + H] ⁺ (Calcd. For C ₁₂ H ₂₂ N ₄ O ₅ Si, MW = 330.14).
Compound D: 5'-O-(i-Propyldimethylsilyl) -5-azacytidine (in formula (I), R ¹ = i-Propyldimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / simple Separation method: Reaction time = Approximately 1 hour, Column elution solvent: Ethyl acetate-methanol system, Isolation yield: 13%)
^{1 1} H-NMR (CDCl ₃ ): 8.56 (s, 1H), 6.81 (br, 1H), 6.08 (br, 1H), 5.85 (d, J = 3.6Hz, 1H), 5.62 (br, 1H), 4.31 -4.33 (m, 1H), 4.24-4.28 (m, 2H), 3.92 (dd, J = 11.6 and 2.4Hz, 1H), 3.76 (dd, J = 11.6 and 2.4Hz, 1H), 3.72 (br, 1H) ), 0.93 (d, J = 6.8Hz, 6H), 0.79-0.88 (m, 1H), 0.07 (s, 3H), and 0.06 (s, 3H).
¹³ C-NMR (CDCl ₃ ): 166.4, 155.6, 155.4, 92.4, 87.0, 71.7, 62.2, 16.8, 16.7, 14.1, -4.7, and -4.8.
Mass: 345.2 [M + H] ⁺ (Calcd. For C ₁₃ H ₂₄ N ₄ O ₅ Si, MW = 344.15).
Compound E: 5'-O- (t-Butyldimethylsilyl) -5-azacytidine (in formula (I), R ¹ = t-Butyldimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / simple Separation method: Reaction time = Approximately 3 hours, Column elution solvent: Ethyl acetate-methanol system, Isolation yield: 12%)
¹ H-NMR (CDCl ₃ ): 8.50 (s, 1H), 6.32 (br, 1H), 5.81 (d, J = 3.6Hz, 1H), 5.76 (br, 1H), 5.45 (br, 1H), 4.35 (d, J = 2.0Hz, 1H), 4.24-4.29 (m, 2H), 3.93 (dd, J = 12.0 and 2.4Hz, 1H), 3.78 (dd, J = 12.0 and 2.0Hz, 1H), 3.54 ( br, 1H), 0.86 (s, 9H), and 0.06 (s, 6H).
¹³ C-NMR (CDCl ₃ ): 167.2, 156.4, 156.0, 93.6, 88.2, 78.1, 72.8, 63.7, 26.5, 18.9, -5.0, and -5.1.
Mass: 359.2 [M + H] ⁺ (Calcd. For C ₁₄ H ₂₆ N ₄ O ₅ Si, MW = 358.17).
Compound F: 5'-O- (Benzyldimethylsilyl) -5-azacytidine (in formula (I), R ¹ = Benzyldimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / isolation method: Reaction Time = about 17 hours, column elution solvent: ethyl acetate-methanol system, isolation yield: 23%)
¹ H-NMR (CDCl ₃ ): 8.45 (s, 1H), 7.19-7.25 (m, 2H). 7.06-7.10 (m, 1H). 6.98-7.00 (m, 2H). 6.18 (br, 1H), 5.77 (d, J = 4.0Hz, 1H), 5.67 (br, 1H), 5.27 (br, 1H), 4.31-4.32 (m, 1H), 4.10-4.16 (m, 2H), 3.84 (dd, J = 8.0 and 2.4Hz, 1H), 3.68 (dd, J = 11.6 and 1.6Hz, 1H), 3.38 (br, 1H), 2.16 (s, 2H), 0.12 (s, 3H), and 0.11 (s, 3H) ..
¹³ C-NMR (CDCl ₃ ): 166.6, 155.9, 155.4, 138.1, 128.5, 128.3, 124.7, 93.1, 87.5, 72.3, 62.5, 26.3, -2.53, and -2.58.
Mass: 393.2 [M + H] ⁺ (Calcd. For C ₁₇ H ₂₄ N ₄ O ₅ Si, MW = 392.15).
Compound G: 5'-O- (n-Octyldimethylsilyl) -5-azacytidine: (In formula (I), R ¹ = n-Octyldimethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions Isolation method: Reaction time = Approximately 1 hour, Column elution solvent: Ethyl acetate-methanol system, Isolation yield: 18%)
¹ H-NMR (CD ₃ OD): 8.78 (s, 1H), 5.79 (d, J = 1.6Hz, 1H), 4.13-4.19 (m, 2H), 4.07 (dt, J = 6.8 and 2.0Hz, 1H) ), 4.03 (dd, J = 12.0 and 2.4Hz, 1H), 3.82 (dd, J = 12.0 and 2.0Hz, 1H), 1.22-1.42 (m, 8H), 0.86-0.93 (m, 4H), 0.62- 0.72 (m, 3H), 0.15 (s, 6H), and 0.14-0.18 (m, 2H).
¹³ C-NMR (CD ₃ OD): 156.6, 156.0, 155.2, 90.8, 84.1, 75.2, 68.5, 60.5, 33.2, 31.8, 29.1, 29.0, 22.9, 22.4, 15.5, 13.1, -3.6, and -3.7.
Mass: 415.4 [M + H] ⁺ (Calcd. For C ₁₈ H ₃₄ N ₄ O ₅ Si, MW = 414.23).
Compound H: 5'-O- (n-Octyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = n-Octyldimethylsilyl group, R = R ² = H) (Synthesis conditions / isolation Method: Reaction time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 24%)
^{1 1} H-NMR (CD ₃ OD): 8.65 (s, 1H), 6.12 (t, J = 5.6Hz, 1H), 4.34-4.37 (m, 1H), 4.00-4.02 (m, 1H), 3.91-3.95 (m, 1H), 3.76-3.79 (m, 1H), 2.45 (ddd, J = 13.6, 6.4, and 4.4Hz, 1H), 2.24 (m, 1H), 1.27-1.34 (m, 8H), 0.87- 0.89 (m, 4H), 0.61-0.63 (m, 3H), and 0.12 (s, 6H).
¹³ C -NMR (CD ₃ OD): 156.2, 155.8, 155.1, 87.9, 86.7, 70.5, 61.8, 41.6, 33.2, 31.8, 29.1, 22.4, 15.6, 13.1, -1.38, -2.96, -3.73, and -3.83 ..
Mass: 399.3 [M + H] ⁺ (Calcd. For C ₁₈ H ₃₄ N ₄ O ₅ Si, MW = 398.23).
Compound I: 5'-O-(t-Butyldiphenylsilyl) -5-azacytidine (in formula (I), R ¹ = t-Butyldiphenylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / simple Separation method: Reaction time = Approximately 2 hours, Column elution solvent: Ethyl acetate-methanol system, Isolation yield: 48%)
¹ H-NMR (CD ₃ OD): 8.63 (s, 1H), 7.69-7.72 (m, 4H), 7.38-7.47 (m, 6H), 5.81 (d, J = 2.4Hz, 1H), 4.32 (dd) , J = 7.2 and 5.2Hz, 1H), 4.23 (dd, J = 5.2 and 2.4Hz, 1H), 4.03-4.09 (m, 2H), 3.82 (dd, J = 11.6 and 2.8Hz, 1H), and 1.08 (s, 9H).
¹³ C-NMR (CD ₃ OD): 166.4, 155.5, 155.0, 135.4, 135.2, 132.5, 132.3, 129.6, 127.5, 90.8, 83.9, 74.7, 68.7, 62.5, and 26.0.
Mass: 483.4 [M + H] ⁺ (Calcd. For C ₂₄ H ₃₀ N ₄ O ₅ Si, MW = 482.20).
Compound J: 5'-O- (Triethylsilyl) -5-azacytidine (in formula (I), R ¹ = Triethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / isolation method: Reaction Time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 10%)
^{1 1} H-NMR (CD ₃ OD): 8.77 (s, 1H), 5.80 (d, J = 2.0Hz, 1H), 4.22 (dd, J = 6.8 and 4.8Hz, 1H), 4.15 (dd, J = 4.8) and 2.0Hz, 1H), 4.03-4.10 (m, 2H), 3.85 (dd, J = 11.6 and 2.0Hz, 1H), 1.00 (t, J = 8.4Hz, 9H), and 0.67-0.74 (m, 6H) ).
¹³ C-NMR (CD ₃ OD): 163.0, 152.3, 151.5, 87.1, 80.4, 71.6, 64.7, 57.1, 2.07, and 0.00.
Mass: 359.2 [M + H] ⁺ (Calcd. For C ₁₄ H ₂₆ N ₄ O ₅ Si, MW = 358.17).
Compound K: 5'-O- (Triethylsilyl) -5-aza-2'-deoxycytidine (R ¹ = Triethylsilyl group, R = R ² = H in formula (I)) (Synthesis conditions / isolation method: Reaction time = Approximately 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 81%)
¹ H-NMR (CDCl ₃ ): 8.62 (s, 1H), 6.26 (t, J = 6.0Hz, 1H), 6.25 (br, 1H), 5.58 (br, 1H), 4.47-4.51 (m, 1H) , 4.09-4.11 (m, 1H), 3.93 (dd, J = 10.8 and 2.4Hz, 1H), 3.82 (dd, J = 11.6 and 2.0Hz, 1H), 2.64-2.70 (m, 1H), 2.66 (br , 1H), 2.23 (dt, J = 12.0 and 6.4Hz, 1H), 0.96 (t, J = 8.0Hz, 9H), and 0.63 (q, J = 8.0Hz, 6H).
¹³ C-NMR (CDCl ₃ ): 166.3, 156.0, 154.1, 87.6, 86.8, 71.6, 62.3, 42.6, 6.7, and 4.1.
Mass: 343.3 [M + H] ⁺ (Calcd. For C ₁₄ H ₂₆ N ₄ O ₄ Si, MW = 342.17).
Compound L: 5'-O- (i-Propyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = i-Propyldimethylsilyl group, R = R ² = H) (Synthesis conditions / isolation Method: Reaction time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 48%)
¹ H-NMR (DMSO-d ₆ ): 8.38 (s, 1H), 7.51 (br s, 1H), 7.49 (br s, 1H), 6.00 (t, J = 6.4Hz, 1H), 5.25 (d, J = 4.8Hz, 1H), 4.16 (q, J = 4.4Hz, 1H), 3.85-3.62 (m, 3H), 2.25-2.03 (m, 2H), 0.92-0.85 (m, 6H), 0.85-0.74 (m, 1H), and 0.02 (s, 6H).
¹³ C-NMR (DMSO-d ₆ ): 166.4, 156.0, 153.6, 87.6, 85.6, 70.4, 62.7, 40.6, 17.3, 14.3, and -4.14.
Mass: 329.4 [M + H] ⁺ (Calcd. For C ₁₃ H ₂₄ N ₄ O ₄ Si, MW = 328.16).
Compound M: 5'-O- (i-Propyldiethylsilyl) -5-azacytidine (in formula (I), R ¹ = i-Propyldiethylsilyl group, R = OR ³ , R ² = R ³ = H) (Synthesis conditions / simple Separation method: Reaction time = Approximately 1 hour, Column elution solvent: Ethyl acetate-methanol system, Isolation yield: 21%)
¹ H-NMR (CDCl ₃ ): 8.56 (s, 1H), 7.04 (br, 1H), 6.21 (br, 1H), 5.85 (d, J = 2.8Hz, 1H), 5.70 (br, 1H), 4.28 (s, 3H), 3.98 (d, J = 11.2Hz, 1H), 3.81 (d, J = 11.2Hz, 1H), 3.79 (br, 1H), 0.93-0.99 (m, 13H), and 0.61-0.65 (m, 4H).
¹³ C-NMR (CDCl ₃ ): 166.4, 155.6, 155.5, 92.2, 87.0, 71.5, 62.5, 17.3, 17.2, 12.5, 7.0, 3.0, and 2.9.
Mass: 373.3 [M + H] ⁺ (Calcd. For C ₁₅ H ₂₈ N ₄ O ₅ Si, MW = 372.18).
Compound N: 5'-O- (i-Propyldiethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = i-Propyldiethylsilyl group, R = R ² = H) (Synthesis conditions / isolation Method: Reaction time = about 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 56%)
¹ H-NMR (DMSO-d ₆ ): 8.42 (s, 1H), 7.55 (br s, 1H), 7.53 (br s, 1H), 6.05 (t, J = 6.4Hz, 1H), 5.30 (d, J = 4.4Hz, 1H), 4.24 (q, J = 4.4Hz, 1H), 3.90-3.72 (m, 3H), 2.29-2.07 (m, 2H), 0.99-0.88 (m, 13H), and 0.68- 0.55 (m, 4H).
¹³ C-NMR (DMSO-d ₆ ): 166.4, 155.9, 153.6, 87.7, 85.6, 70.4, 63.0, 40.6, 17.6, 12.5, 7.34, and 3.00.
Mass: 357.4 [M + H] ⁺ (Calcd. For C ₁₅ H ₂₈ N ₄ O ₄ Si, MW = 356.19).
Compound O: 5'-O- (Cyclopentyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = Cyclopentyldimethylsilyl group, R = R ² = H) (Synthesis conditions / isolation method: Reaction time = Approximately 0.5 hours, column elution solvent: ethyl acetate-methanol system, isolation yield: 13%)
¹ H-NMR (DMSO-d ₆ ): 8.39 (s, 1H), 7.49 (s, 2H), 6.00 (t, J = 6.4Hz, 1H), 5.25 (d, J = 4.8Hz, 1H), 4.15 (q, J = 4.0Hz, 1H), 3.86-3.63 (m, 3H), 2.02-2.24 (m, 2H), 1.17-1.72 (m, 8H), 0.94 (dq, J = 8.4 and 2.0Hz, 1H ), and 0.01 (s, 6H).
¹³ C-NMR (DMSO-d ₆ ): 166.4, 156.0, 153.6, 87.6, 85.7, 70.5, 62.7, 40.4, 27.5, 27.2, 25.7, and -3.2.
Mass: 355.5 [M + H] ⁺ (Calcd. For C ₁₅ H ₂₆ N ₄ O ₄ Si, MW = 354.17).
Compound P: 5'-O- (Cyclohexyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = Cyclohexyldimethylsilyl group, R = R ² = H) (Synthesis conditions / isolation method: Reaction time = Approximately 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 53%)
¹ H-NMR (DMSO-d ₆ ): 8.37 (s, 1H), 7.49 (s, 2H), 5.99 (t, J = 6.2Hz, 1H), 5.24 (d, J = 4.8Hz, 1H), 4.15 (q, J = 4.4Hz, 1H), 3.62-3.85 (m, 3H), 2.03-2.25 (m, 2H), 1.52-1.70 (m, 5H), 0.96-1.20 (m, 5H), 0.64 (dt , J = 12.4 and 3.2Hz, 1H), and 0.00 (s, 6H).
¹³ C-NMR (DMSO-d ₆ ): 166.4, 156.0, 153.6, 87.7, 85.8, 70.5, 62.8, 40.4, 27.8, 26.9, 26.8, 26.3, and -3.73.
Mass: 369.4 [M + H] ⁺ (Calcd. For C ₁₆ H ₂₈ N ₄ O ₄ Si, MW = 368.19).
Compound Q: 5'-O- (Cyclopentyldiethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ¹ = Cyclopentyldiethylsilyl group, R = R ² = H) (Synthesis conditions / isolation method: Reaction time = Approximately 1 hour, column elution solvent: ethyl acetate-methanol system, isolation yield: 31%)
¹ H-NMR (DMSO-d ₆ ): 8.43 (s, 1H), 7.55 (s, 2H), 6.05 (t, J = 6.2Hz, 1H), 5.30 (d, J = 3.6Hz, 1H), 4.23 (br s, 1H), 3.69-3.92 (m, 3H), 2.04-2.35 (m, 2H), 1.25-1.83 (m, 8H), 0.83-1.15 (m, 6H), 1.04 (m, 1H), and 0.61 (q, J = 4.0Hz, 4H).
¹³ C-NMR (DMSO-d ₆ ): 166.5, 155.9, 153.6, 87.8, 85.7, 70.5, 63.1, 41.5, 27.6, 27.1, 24.0, 7.4, and 3.9.
Mass: 383.3 [M + H] ⁺ (Calcd. For C ₁₇ H ₃₀ N ₄ O ₄ Si, MW = 382.20).

3'Position Cyriloxy-5-aza-2'-Deoxycytidine Synthesis Method:



3', 5'-disilyloxy-5-aza-2'-deoxycytidines ( ^{a compound in (I) in which R is a hydrogen atom and R 1} and R ² are silyl groups) (1 mM) anhydrous cyclopentyl Camphorsulfonic acid (1 mM) is added to a methyl ether solution (10 mL), and the mixture is stirred at room temperature for about 1 day. After neutralizing the reaction solution with a baking soda solution, the insoluble matter is filtered off, the filtrate is dried under reduced pressure, and the oily residue obtained is separated by a silica gel pack column (Yamazen Smart Flash MS system device). By purification, the desired 3'position silyloxy-5-aza-2'-deoxycytidine derivative (in formula (I), R and R ¹ are hydrogen atoms and R ² is a silyl group). Can be isolated as a white powder. This synthesis method is hereinafter abbreviated as synthesis method A.
Further, in the above 3', 5'-disilyloxy-5-aza-2'-deoxycytidines (in (I), R is a hydrogen atom and R ¹ and R ² are silyl groups.) (1 mM). ), A catalytic amount of CAN (Cerium Ammonium Nitrate) is added to the anhydrous isopropanol solution (10 mL), and the mixture is stirred at room temperature for about 1 day to obtain the desired 3'position silyloxy-5-aza-2'-. A deoxycytidine derivative (in the formula (I) ^{, a compound in which R and R 1} are hydrogen atoms and R ² is a silyl group) can be obtained. This synthesis method will be abbreviated as synthesis method B hereafter.
Compound R: 3'-O- (Triethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ² = Triethylsilyl group, R = R ¹ = H) (Synthesis conditions / Isolation method: Reaction time = Approximately 1 day, column elution solvent: ethyl acetate-methanol system, isolation yield: 29% (synthesis method A), 50% (synthesis method B))
¹ H-NMR (DMSO-d ₆ ): 8.48 (s, 1H), 7.54 (br s, 1H), 7.51 (br s, 1H), 6.01 (t, J = 6.4Hz, 1H), 5.10 (t, J = 5.2Hz, 1H), 4.39-4.42 (m, 1H), 3.81 (q, J = 3.6Hz, 1H), 3.57-3.63 (m, 1H), 3.51-3.55 (m, 1H), 2.15-2.28 (m, 2H), 0.92 (t, J = 8.4Hz, 9H), and 0.58 (q, J = 7.6Hz, 6H).
¹³ C-NMR (DMSO-d ₆ ): 165.8, 155.9, 153.1, 87.7, 85.1, 71.3, 60.6, 40.7, 6.6, and 4.1.
Mass: 343 [M + H] ⁺ (Calcd. For C ₁₄ H ₂₆ N ₄ O ₄ Si, MW = 342.17).
Compound S: 3'-O- (n-Propyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ² = n-Propyldimiethylsilyl group, R = R ¹ = H) (Synthetic conditions / isolation Method: Reaction time = about 1.5 hours, column elution solvent: ethyl acetate-methanol system, isolation yield: 10% (synthesis method A))
¹ H-NMR (DMSO-d ₆ ): 8.48 (s, 1H), 7.53 (br s, 1H), 7.51 (br s, 1H), 5.99 (t, J = 6.4Hz, 1H), 5.09 (br s) , 1H), 4.38-4.41 (m, 1H), 3.79 (q, J = 4.0Hz, 1H), 3.60 (br d, J = 12.4Hz, 1H), 3.51 (br d, J = 12.4Hz, 1H) , 2.14-2.25 (m, 2H), 1.29-1.37 (m, 2H), 0.94 (t, J = 7.2Hz, 3H), 0.56-0.60 (m, 2H), and 0.10 (s, 6H).
¹³ C-NMR (DMSO-d ₆ ): 165.9, 155.9, 153.1, 87.5, 85.1, 71.1, 60.5, 18.6, 17.9, 16.2, and -1.6.
Mass: 329 [M + H] ⁺ (Calcd. For C ₁₃ H ₂₄ N ₄ O ₄ Si, MW = 328.16).
Compound T: 3'-O- (i-Propyldimethylsilyl) -5-aza-2'-deoxycytidine (in formula (I), R ² = i-Propyldimiethylsilyl group, R = R ¹ = H) (Synthetic conditions / isolation Method: Reaction time = about 3 hours, column elution solvent: ethyl acetate-methanol system, isolation yield: 17% (synthesis method A))
¹ H-NMR (DMSO-d ₆ ): 8.48 (s, 1H), 7.53 (br s, 1H), 7.51 (br s, 1H), 6.01 (t, J = 6.4Hz, 1H), 5.09 (t, J = 5.2Hz, 1H), 4.39-4.42 (m, 1H), 3.80 (q, J = 3.6Hz, 1H), 3.58-3.81 (m, 1H), 3.51-3.55 (m, 1H), 2.17-2.26 (m, 2H), 0.93 (br s, 3H), 0.92 (br s, 3H), 0.81-0.86 (m, 1H), and 0.07 (s, 6H).
¹³ C-NMR (DMSO-d ₆ ): 165.9, 155.9, 153.1, 87.5, 85.0, 71.2, 60.5, 40.5, 16.7, 13.8, -3.9, and -4.0.
Mass: 329 [M + H] ⁺ (Calcd. For C ₁₃ H ₂₄ N ₄ O ₄ Si, MW = 328.16).

Test Example 1

Stability of 5-azacitidine sugar moiety silyl ether derivative to cytidine deaminase 5-Azacitidine sugar moiety silyl ether derivative (see formula (I)) About 1 mg was dissolved in 1 mL of acetonitrile, and 10 μL thereof was added to 1 mL of PBS. 10 μL of a PBS solution of cytidine deaminase was added to the obtained solution, and the mixture was stirred at 37 ° C. for about 1 hour. 1 mL of acetonitrile was added to the reaction solution, and the mixture was centrifuged, and the supernatant was analyzed by HPLC. For example, 5'-O-(t-Butyldimethylsilyl) -5-azacytidine (Compound E), 5'-O- (Triethylsilyl) -5-azacytidine (Compound J), 5'-O- (Triethylsilyl) -5-aza Table 1 shows the analysis results in the case of -2'-deoxycytidine (Compound K).
Cytidine deaminase: CDA (1-146aa), Human, His-tagged, Recombinant cytidine deaminase (ATGen)
HPLC measurement conditions:
Column: CAPCELL PAK ADME
4.6 mm x 150 mm, particle size: 3 μm
Elution: Eluate A = 10 mM purified water containing ammonium formic acid
Eluent B = acetonitrile
A: B = 99: 1 → 5:95, gradient mode for 30 minutes Outflow rate: 1.0 mL / min Oven temperature: 40 ° C
Detector: UV240nm

As described above, the 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) according to the present invention was very stable to cytidine deaminase. On the other hand, 5-azacitidine and 5-aza-2'-deoxycytidine were completely eliminated under the above reaction conditions in both cases.

Test Example 2

5-Non-enzymatic hydrolysis of azacitidine sugar silyl ether derivatives 5-Azacitidine sugar silyl ether derivatives (see formula (I)), eg, 5'-O- (Triethylsilyl) -5-azacytidine (Compound J) ) About 1 mg was dissolved in 1 mL of acetonitrile, 5 μL of it was added to 100 μL of a 10 mM PBS solution, and the mixture was stirred at 37 ° C. As a result of HPLC analysis of the reaction product over time, the formation of 5-azacitidine (in formula (I), R ¹ , R ² and R ³ are hydrogen atoms) was confirmed, and the formation of other decomposition products was confirmed. Was hardly recognized. Similar results were obtained with 5'-O- (Triethylsilyl) -5-aza-2'-deoxycytidine (Compound K), and the corresponding desilylated form (5-Aza-2'-deoxycytidine: formula (5-Aza-2'-deoxycytidine) In I), ^{the formation of R 1} , R ² and R are hydrogen atoms) was confirmed.
The HPLC measurement conditions are the same analytical conditions as in Test Example 1.

Test Example 3

Anti-ATL activity of 5-azacitidine sugar silyl ether derivative RPMI-1640 (* ATN-) containing 10% FBS and 1% Penn-Strep in the culture medium of the ATL cell line (HTLV-1 infected cell line) in the table below. Inoculate an additional 1% NEAA for one strain and an additional 50 ng / ml h IL-2 for the ILT-Mat strain)) at 3,000 to 7,000 cells / 50 ml / well on a 96 well plate, 5 Incubated at 37 ° C for about 3 hours under a stream of% carbon dioxide. Next, 50 ml of each compound diluted with the culture solution to 100 mM to 20 nM was added to the above 96-well plate, incubated for 48 hours, and then 10 ml of the compound diluted with the culture solution was added to the final concentration. It was re-added so that it would be the same. After further incubating for 48 hours (96 hours in total), react using CCK-8 reagent (DOJINDO, CK04) according to the attached manual, and then use a plate reader (Varioskan Flash, Thermo Fisher Scientific Co., Ltd.) to 450 nm for each well. And 620 nm (Blank) absorption was measured. _{The IC 50} (nM) value was calculated by expressing the value of each sample processing well when the value of the sample-free well was set to 100% as a relative value (see Table 3).

ATL cell line (HTLV-1 infected cell line): ATN-1, ILT-Mat, TL-Mor line purchased from RIKEN BioResource Center (RIKEN BRC), MJ line purchased from American Type Culture Collection (ATCC), MT- 2, MT-4 strain was purchased from JCRB cell bank.
From the results in Table 3, it was found that the sugar moiety silyl ether derivative of 5-aza-2'-deoxycytidine has a very high anti-ATL activity.

Test Example 4

5-DNA demethylation effect of azacitidine sugar silyl ether derivative The CpG island cytosine methylation ratio of LINE-1 (Long interspersed nucleotide factor-1) was measured using the pyrosequencing method.
Add 100 nM or 500 nM concentration of 5-azacitidine sugar moiety silyl ether derivative solution to ATL cell / MT-2 (about 50,000 cells / mL) -containing solution, and RPMI-1640 (containing 10% FBS and Penn-strep) medium. After incubating in the medium for 48 hours, the 5-azacitidine sugar moiety silyl ether derivative solution was added again so that the final concentration became 100 nM or 500 nM, and in RPMI-1640 (containing 10% FBS and Penn-strep) medium. Incubated for 48 hours (96 hours total). Then, DNA was extracted from the cells, treated with bisulfite, and then the CpG methylation rate of LINE-1 was measured (see Table 4).

As a result, it was confirmed that the 5-azacitidine sugar moiety silyl ether derivative (see formula (I)) has a DNA demethylation effect on MT-2.

According to the present invention, a DNMT inhibitor having high stability against the metabolic enzyme cytidine deaminase can be provided to the medical field as a novel ATL therapeutic agent or preventive agent.

Claims (11)

Equation (I):


(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ATL prophylactic / therapeutic agent containing the compound represented by) or a salt thereof. Prevention of ATL containing the compound according to claim 1 or a salt thereof, wherein R ¹ is a silyl group represented by the formula (II) and R ² and R or R ^{3 are hydrogen atoms.} Therapeutic agent. Prevention of ATL containing the compound according to claim 1 or a salt thereof, wherein R ² is a silyl group represented by the formula (II) and R ¹ and R or R ^{3 are hydrogen atoms.} Therapeutic agent. Prevention of ATL containing the compound according to claim 1 or a salt thereof, wherein R ¹ and R ² are silyl groups represented by the formula (II), respectively, and R or R ^{3 is a hydrogen atom.}・ Therapeutic agent. The prophylactic / therapeutic agent for ATL containing the compound according to claim 1 or a salt thereof, wherein R ¹ is a hydrogen atom and R ² and R ^{3 are silyl groups represented by the formula (II), respectively.} The prophylactic / therapeutic agent for ATL containing the compound according to claim 1 or a salt thereof, wherein R ¹ , R ² and R ^{3 are silyl groups represented by the formula (II), respectively.} Claimed that R ⁴ , R ⁵ and R ⁶ _{are C 1 to} C ₈ alkyl groups or C _{6 to} C ₁₀ aryl groups or C _{7 to} C ₁₄ aryl alkyl groups, respectively, which may have substituents. The preventive / therapeutic agent for ATL according to 1. The prophylactic / therapeutic agent for ATL according to claim 7, wherein the C _{6 to} C _{10 aryl groups are phenyl groups or naphthyl groups.} The prophylactic / therapeutic agent for ATL according to claim 7, wherein the C _{7 to} C ₁₄ arylalkyl group is a benzyl group, a phenethyl group or a naphthylmethyl group. Equation (I):


(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ), A method for preventing or treating ATL, which comprises administering an effective amount of a compound or a salt thereof. Formula (I) for producing a pharmaceutical composition for the prevention or treatment of ATL:

(In the formula, R is an OR ³ group or a hydrogen atom, and R ¹ , R ² and R ³ are a hydrogen atom or a hydrogen atom, respectively:

(Wherein, R ^4, R ⁵ and R ⁶ each have a substituent is also an alkyl group or an aryl group or an arylalkyl group.) Is a silyl group represented by. However, this ^{excludes cases where R 1} , R ² and R ³ are hydrogen atoms at the same time. ) Or a salt thereof.