WO2020090698A1

WO2020090698A1 - Pharmaceutical composition

Info

Publication number: WO2020090698A1
Application number: PCT/JP2019/042080
Authority: WO
Inventors: 孫市酒向; 晋平杉山; 祐樹倉橋; 弘臣脇田; 晋也木村; 達郎渡邉; 博志嬉野
Original assignee: 大原薬品工業株式会社; 国立大学法人佐賀大学
Priority date: 2018-10-30
Filing date: 2019-10-28
Publication date: 2020-05-07
Also published as: JP7228169B2; CN112638422A; JPWO2020090698A1; CN112638422B

Abstract

Provided, as a cancer prophylactic drug or a cancer therapeutic drug, are a compound having the effect of inhibiting DNA methyltransferase (DNMT), and another drug.

Description

Pharmaceutical composition

The present invention relates to a preventive or therapeutic drug for cancer.

Blood cancer refers to a type of cancer that attacks the blood, bone marrow, and / or lymphatic system. This type of cancer includes leukemia and multiple myeloma, all of which can be deadly diseases that require new, more effective treatments.

ATL (Adult T-cell Leukemia / Lymphoma: Adult T-cell leukemia / lymphoma) is a disease discovered and named by Takatsuki et al. (Non-patent Documents 1 and 2) in 1976, and the oncogenic virus HTLV-1. (Human T-cell Leukemia / Lymphotropic Virus type-1) is a leukemia or malignant lymphoma caused by infection. The number of HTLV-1 infected people is high in the southern part of Japan (particularly Kyushu and Okinawa), and is localized in the Caribbean coastal countries, Central Africa and South America.
The routes of HTLV-1 infection include breastfeeding, sexual intercourse, blood transfusion, etc., and the carriers are said to be 5 to 10 million worldwide (about 1.1 million in Japan). Six to seven percent of them, and two to three percent of women, develop ATL each year. Note that the period from HTLV-1 infection to the onset of ATL is very long, and as a result, many elderly carriers have ATL onset (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 treatment of ATL, but recurrence or drug resistance In many cases, standard treatment methods have not yet been established.
Recently, a new chemokine receptor CCR4 antibody drug, Mogamulizumab (product name: poteridio) (non-patent document 7), an antiviral drug, abacavir (non-patent document 8), histone deacetylase as a new therapeutic agent for ATL (HDAC) inhibitor Vorinostat (product name: Zolinza) (Non-Patent Document 9) and the possibility of dual inhibitor DS-3201b of histone methyltransferase EZH1 / 2 have attracted attention.

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

Also, DNMTs are closely related to cancer development. That is, it is considered that 60 to 90% of all CpGs are methylated at the 5-position of the cytosine ring, but abnormal levels of DNA methylation are closely related to the regulation of gene expression, and the promoter region ( It has been clarified that the transcription / expression of a gene in which the 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, the cell is equipped with a mechanism to introduce a methyl group into the 5-position of the cytosine ring at the same position as the parent chain in the newly created DNA chain, which enables this "replication of DNA methylation". DNMTs. Therefore, in cancerous cells, most of the tumor suppressor genes are suppressed in transcription / expression, and become in a silencing state, and are in a state in which they easily proliferate.

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

As a selective enzyme inhibitor against DNMTs having such a background, 5-azacytidine (product name: "Vidaza (registered trademark)") and its 2'-deoxy form (decitabine, product name: "Dakogen (registered trademark)" ) ”) Was found and is clinically used as a therapeutic drug for high-risk myelodysplastic syndrome and acute myeloid leukemia. The chemical structure of these drugs is very similar to that of cytosine nucleosides (structure in which the carbon atom at the 5-position of the cytosine ring is replaced by a nitrogen atom), and the drug undergoes a nucleic acid biosynthetic route to produce DNA instead of 2'-deoxycytidine. By invading into the tumor suppressor gene region (CpG island), DNMTs suicide-mediated methylation reaction of cytosine ring position 5 is suicidally inhibited, and normal expression of the tumor suppressor gene is enabled to exert therapeutic effect. It is said to be forgotten.

However, no effective drug has been found for blood cancer yet.

Due to the above circumstances, there is a demand for a drug that overcomes blood cancer in clinical settings.

An object of the present invention is to provide a novel preventive or therapeutic drug for cancer (especially blood cancer). ‥

The present inventors have conducted intensive research to find out an excellent prophylactic or therapeutic drug for blood cancer, and as a result, a pharmaceutical composition comprising a DNMT inhibitor (particularly compound (I)) and another drug in combination. It was found that the product is useful for the prevention or treatment of blood cancer.
The present invention has been completed based on these findings.

That is, the present invention has solved the above problems by providing the inventions [1] to [29] described below.
[1]
A pharmaceutical composition comprising a DNMT inhibitor in combination with other agents.
[2]
A pharmaceutical composition comprising another drug for use in combination with a DNMT inhibitor.
[3]
A pharmaceutical composition comprising a DNMT inhibitor and other agents.
[4]
The DNMT inhibitor has the formula (I):

(In the formula, R and R ′ are each an OR ³ group, a hydrogen atom, a halogen atom or an alkyl group, and R ¹ , R ² and R ³ are each a hydrogen atom or a formula (II):

(In the formula, R ⁴ , R ⁵ and R ⁶ are each an alkyl group, an aryl group or an arylalkyl group which may have a substituent). ) Or a salt thereof, decitabine, azacitidine, RG108, thioguanine, Zebularine, SGI-110, CC-486, SGI-1027, lomeguatrib or procainamide hydrochloride, any of [1] to [3] The pharmaceutical composition according to the item 1.
[5]
The pharmaceutical composition according to [4], wherein R ¹ is a silyl group represented by formula (II), and R ² and R or R ³ are hydrogen atoms.
[6]
The pharmaceutical composition according to [4], wherein R ² is a silyl group represented by formula (II), and R ¹ and R or R ³ are hydrogen atoms.
[7]
The pharmaceutical composition according to [4], wherein each of R ¹ and R ² is a silyl group represented by the formula (II), and each of R and R ³ is a hydrogen atom.
[8]
The pharmaceutical composition according to [4], wherein R ¹ is a hydrogen atom, and R ² and R ³ are each a silyl group represented by the formula (II).
[9]
The pharmaceutical composition according to [4], wherein each of R ¹ , R ² and R ³ is a silyl group represented by the formula (II).
[10]
The pharmaceutical composition according to [4], wherein each of R ¹ , R ² and R ³ is a hydrogen atom.
[11]
R ⁴ , R ⁵ and R ⁶ are each a C ₁ -C ₈ alkyl group which may have a substituent, a C ₆ -C ₁₀ aryl group or a C ₇ -C ₁₄ arylalkyl group which may have a substituent. ] The pharmaceutical composition of description.
[12]
The pharmaceutical composition according to [11], wherein the C ₆ -C ₁₀ aryl group is a phenyl group or a naphthyl group.
[13]
The pharmaceutical composition according to [11], wherein the C ₇ -C ₁₄ arylalkyl group is a benzyl group, a phenethyl group or a naphthylmethyl group.
[14]
The pharmaceutical composition according to any one of [1] to [13], wherein the other drug is an enzyme inhibitor.
[15]
The enzyme inhibitor is selected from the group consisting of histone acetylase inhibitors, histone deacetylase inhibitors, histone methylase inhibitors, histone demethylase inhibitors and ribonucleoside diphosphate reductase inhibitors The pharmaceutical composition according to [14], which is one or more kinds.
[16]
The pharmaceutical composition according to [14], wherein the enzyme inhibitor is one or more selected from the group consisting of DS-3201b, HBI-8000, trichostatin A (TSA), suramin, EPZ005687 and Adox. ..
[17]
The pharmaceutical composition according to any one of [1] to [16], further comprising a pharmaceutically acceptable carrier.
[18]
The pharmaceutical composition according to any one of [1], [2] and [4] to [17], wherein the DNMT inhibitor and the other drug are administered simultaneously.
[19]
The pharmaceutical composition according to any one of [1], [2] and [4] to [17], wherein the DNMT inhibitor and the other drug are administered separately.
[20]
The pharmaceutical composition according to any one of [1], [2] and [4] to [17], wherein the DNMT inhibitor is administered before administration of the other drug.
[21]
The pharmaceutical composition according to any one of [1], [2] and [4] to [17], wherein the DNMT inhibitor is administered after administration of another drug.
[22]
The pharmaceutical composition according to any one of [1] to [21], which is used for preventing or treating cancer.
[23]
The pharmaceutical composition according to [22], wherein the cancer is leukemia.
[24]
The pharmaceutical composition according to [23], wherein leukemia is ATL.
[25]
A preventive or therapeutic drug for cancer, which comprises the pharmaceutical composition according to any one of [1] to [21].
[26]
The preventive or therapeutic drug for cancer according to [25], wherein the cancer is leukemia.
[27]
The preventive or therapeutic drug for cancer according to [26], wherein leukemia is ATL.
[28]
A kit for preventing or treating cancer, which comprises the DNMT inhibitor described in any one of [1] to [22] and another drug.
[29]
A method for preventing or treating cancer in a mammal, which comprises administering an effective amount of a DNMT inhibitor and an effective amount of another drug to the mammal.

The pharmaceutical composition of the present invention is useful as a prophylactic or therapeutic agent for leukemia such as ATL whose expression is caused by DNMT.

Unless otherwise stated, the terms used in the present specification and claims have the meanings described below.

DNMT Inhibitor The “DNA methyltransferase inhibitor” of the present invention is not particularly limited, but for example, the following formula (I):

(In the formula, R ⁴ , R ⁵ and R ⁶ are each an alkyl group, an aryl group or an arylalkyl group which may have a substituent). ) Or a salt thereof, decitabine, azatidine, RG108, thioguanine, Zebularine, SGI-110, CC-486, SGI-1027, lomeguatrib and procainamide hydrochloride. R and R ′ in the formula (I) are preferably an OR ³ group, a hydrogen atom or a halogen atom.

Examples of preferred combinations of R and R'in formula (I) are as follows.
(i) R is an OR3 group, a hydrogen atom, a halogen atom or an alkyl group, and R'is a hydrogen atom.
(ii) R is an OR3 group or a hydrogen atom, and R'is a hydrogen atom.
(iii) R is a fluorine atom and R'is a fluorine atom.

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, unless otherwise specified, and examples thereof include a methyl group and an ethyl group. , Propyl group, isopropyl group, butyl group, sec-butyl group, isobutyl group, tert-butyl group, pentyl group, hexyl group, and other C ₁ -C ₆ alkyl groups, heptyl group, 2-methylhexyl group, 5-methyl group 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-methylheptyl 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 a C ₁ -C ₆ alkyl group is preferable. Preferred examples of C ₁ -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.

“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" refers to an alkyl group substituted with aryl. Preferred is a C ₇ -C ₁₄ arylalkyl group. Examples of C ₇ -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, an aryl group which may have a substituent or an arylalkyl group which may have a substituent” may or may not have a substituent. Means that it may be a substitution. When it is substituted, the substituent may have 1 to 5, preferably 1 to 3 at the substitutable position of the alkyl group, aryl group or arylalkyl group, and the number of substituents is 2 or more. In this case, 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.

“Halogen atom” refers to a fluorine atom, a chlorine atom, a bromine atom, an iodine atom and the like, and preferable examples are a fluorine atom and a chlorine atom.

The salt of the compound represented by formula (I) of the present invention may be any salt as long as it is a pharmacologically acceptable salt. Examples of the salt include inorganic acid salts (eg, hydrochloride, sulfate, hydrobromide, phosphate, etc.), organic acid salts (eg, acetate, trifluoroacetate, succinate, Acid addition salts such as maleate, fumarate, propionate, citrate, tartrate, lactate, oxalate, methanesulfonate, p-toluenesulfonate, etc., and the like. 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. The crystal can be produced by applying a crystallization method known per se to crystallize.

Further, the compound represented by the formula (I) of the present invention may be a solvate (for example, a hydrate), and a solvate and a non-solvate (for example, a non-hydrate). All of the above are included in the compound represented by the formula (I).

Method for Producing Compound Represented by Formula (I) of the Present Invention The 5-azacytidine sugar moiety silyl ether derivative according to the present invention (see Formula (I)) should be produced according to the method shown in WO2017 / 183217. You can

Other agents Examples of other agents of the present invention include enzyme inhibitors, preferably histone deacetylase inhibitors, histone acetylase inhibitors, histone methylase inhibitors, histone demethylation. One or more enzyme inhibitors selected from the group consisting of enzyme inhibitors and ribonucleoside diphosphate reductase inhibitors.

The “histone acetylase inhibitor” is not particularly limited, but examples thereof include C646, MG149, Remodelin, and Anacardic Acid.

The "histone deacetylase inhibitor" is not particularly limited, and examples thereof include vorinostat (SAHA, MK0683), entinostat (MS-275), panobinostat (LBH589), trichostatin A (TSA), mosetinostat (MGCD0103). ), BG45, BRD73954, Berinosutatto (PXD101), romidepsin (FK228, Deshipepuchido), 4SC-202, HPOB, LMK-235, CAY10603, Tasukinimodo, TMP269, Nexturastat A, Rocilinostat (ACY-1215), RGFP966, RG2833 (RGFP109) , Scriptaid, Tubastatin A, Pracinostat (SB939), CUDC-101, M344, CI-34051, dasinostat (LAQ824), tubastatin A hydrochloride, abexinostat (PCI-24781), CUDC-907, AR-42, sodium phenylbutyrate, resminostat, tubacin, xinostat (JNJ-264881585) dihydrochloride Salt, MC1568, Givinostat (ITF2357), Droxinostat, Chimamide (C S055, HBI-8000), CHR-2485, CHR-3996, DAC-060, FRM-0334 (EVP-0334), MGCD-290, CXD-101 (CXD-101). AZD-9468), CG200745, arginine butyrate, sulforaphane, SHP-141, CUDC-907, YM753 (OBP-801), sodium valproate, Pishijin and CI994 (Tacedinaline), and the like.

The “histone methyltransferase inhibitor” is not particularly limited, but examples thereof include DS-3201b, EPZ-6438, GSK-126, Chaetocin, and IX01294.

The “histone demethylase inhibitor” is not particularly limited, and examples thereof include GSKJ4HCl, OG-L002, JIB-04, IOX1, SP2509, ORY-1001 (RG-6016), GSKJ1, ML324, GSK. -LSD1 and 2HCl and the like.

The “ribonucleoside diphosphate reductase inhibitor” is not particularly limited, and examples thereof include gemcitabine, ara C, fludarabine and the like.

The other drug is particularly preferably one or more selected from the group consisting of DS-3201b, HBI-8000, trichostatin A (TSA), suramin, EPZ005687 and Adox.

The pharmaceutical composition can be produced by a method conventionally used in the field of pharmaceutical preparation, such as the method described in the Japanese Pharmacopoeia.
The dosage form for oral administration of the pharmaceutical preparation of the present invention includes solid preparations such as tablets, capsules, granules and powders. In addition, examples of the dosage form for parenteral administration such as intravenous, subcutaneous and intramuscular include injections, suppositories, sublingual tablets and the like. The dosage form for sublingual, subcutaneous or intramuscular administration includes, for example, sustained-release preparations such as sublingual tablets and microcapsules.

As the pharmaceutically acceptable carrier, for example, various organic or inorganic carrier substances conventionally used as a formulation material are used, and an excipient, a lubricant, a binder, a disintegrating agent, a thickener in a solid formulation; a liquid formulation The solvent, the dispersant, the solubilizing agent, the suspending agent, the isotonicity agent, the buffering agent, the soothing agent, etc. are appropriately mixed in appropriate amounts. If necessary, additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used according to a conventional method.

Suitable examples of excipients include lactose, sucrose, D-mannitol, starch, crystalline cellulose, light anhydrous silicic acid and the like. Preferable examples of the lubricant include magnesium stearate, calcium stearate, talc, colloidal silica and the like. Preferable examples of the binder include crystalline cellulose, sucrose, D-mannitol, dextrin, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, polyvinylpyrrolidone and the like. Preferable examples of the disintegrant include starch, carboxymethyl cellulose, carboxymethyl cellulose calcium, croscarmellose sodium, carboxymethyl starch sodium and the like. Preferable examples of the thickener include natural gums, cellulose derivatives, acrylic acid polymers and the like. Preferable examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like. Preferable examples of the dispersant include Tween 80, HCO 60, polyethylene glycol, carboxymethyl cellulose, sodium alginate and the like. Preferable examples of the solubilizing agent include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like. Preferable examples of the suspending agent include stearyl triethanolamine, sodium lauryl sulfate, lauryl aminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate and the like. Preferable examples of the surfactant include hydrophilic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, sodium carboxymethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose. Preferable examples of the tonicity agent include sodium chloride, glycerin, D-mannitol and the like. Preferable examples of the buffer include phosphate, acetate, carbonate, citrate and the like. Preferable examples of soothing agents include benzyl alcohol and the like. Preferable preservatives include, for example, paraoxybenzoic acid esters, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like. Preferable examples of the antioxidant include sulfite, ascorbic acid and the like.

The pharmaceutical composition of the present invention is used for preventing or treating cancer.
Cancers of the head and neck, breast cancer, lung cancer, colon cancer, ovarian cancer, prostate cancer, glioma, glioblastoma, astrocytoma, glioblastoma multiforme , Banayan-Zonana syndrome, Cowden disease, Lermitt-Dachros disease, inflammatory breast cancer, Wilms tumor, Ewing sarcoma, rhabdomyosarcoma, ependymoma, medulloblastoma, kidney cancer, liver cancer, melanoma, pancreatic cancer , Sarcoma, osteosarcoma, giant cell tumor of bone, thyroid, lymphoblastic T cell leukemia, ATL (Adult T-cell Leukemia / Lymphoma), chronic myelogenous leukemia, chronic lymphocytic leukemia, hairy Cell leukemia, acute lymphoblastic leukemia, acute myelogenous leukemia, AML, chronic neutrophil leukemia, acute lymphoblastic T cell leukemia, plasmacytoma, immunoblastic large cell leukemia, mantle cell leukemia, multiple Myeloma megakaryoblastic leukemia, multiple Emerging myeloma, acute megakaryocytic leukemia, promyelocytic leukemia, erythroleukemia, malignant lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, lymphoblastic T cell lymphoma, Burkitt lymphoma, follicular lymphoma, neuroblastoma , Bladder cancer, urothelial cancer, vulvar cancer, cervical cancer, endometrial cancer, renal cancer, mesothelioma, esophageal cancer, salivary gland cancer, hepatocellular carcinoma, gastric cancer, nasopharynx Cancer, cheek cancer, oral cancer, GIST (gastrointestinal stromal tumor), testicular cancer and the like can be mentioned, preferably lymphoblastic T cell leukemia, ATL (Adult T-cell Leukemia / Lymphoma: adult T cell leukemia / lymphoma), chronic myelogenous leukemia, chronic lymphocytic leukemia, hairy cell leukemia, acute lymphoblastic leukemia, acute myelogenous leukemia, AML, chronic neutrophilic leukemia, acute lymphoblastic T cell leukemia , Immunoblastic large cell leukemia A mantle cell leukemia, megakaryoblastic leukemia, acute megakaryocyte leukemia, promyelocytic leukemia, erythroleukemia, most preferably, is a ATL, but is not limited thereto.

When the preventive or therapeutic agent for ATL of the present invention is combined with another drug, the timing of administration of the preventive or therapeutic agent for ATL and the other drug is not limited, and both of the other drugs are simultaneously administered to the administration subject. Alternatively, the doses may be administered at staggered times. The preventive or therapeutic drug for ATL and the other drug may be separately formulated, or a mixture of the two may be mixed. The dose of the other drug may be in accordance with the dose clinically used, and can be appropriately selected depending on the administration subject, administration route, disease, combination and the like. The dose of the other drug may be, for example, 1/3 to 3 times the dose when the other drug is used as a single agent.

The administration form of the preventive or therapeutic agent for ATL of the present invention and the other drug is not particularly limited, and the preventive or therapeutic agent for ATL and the other drug may be combined at the time of administration. Examples of such a dosage form include (1) administration of a single preparation obtained by simultaneously formulating a prophylactic or therapeutic drug for ATL and another drug, (2) prophylactic or therapeutic drug for ATL and other Simultaneous administration of two preparations obtained by separately formulating a drug with the same administration route, (3) Preparation of two preparations obtained by separately preparing a prophylactic or therapeutic drug for ATL and another drug (4) Simultaneous administration of two kinds of preparations obtained by separately formulating a prophylactic or therapeutic drug for ATL and another drug by different administration routes, (5) Administration of two kinds of preparations obtained by separately formulating a prophylactic or therapeutic drug for ATL and another drug with different administration routes (for example, prophylactic or therapeutic drug for ATL → other drugs in this order Administration, or administration in the reverse order) and the like.

By combining the preventive or therapeutic agent for ATL of the present invention with another drug, the following excellent effects can be obtained.
(1) The dose can be reduced as compared with the case where the preventive or therapeutic drug for ATL and another drug are administered alone,
(2) Other drug types can be selected according to the patient's symptoms (mild, severe, etc.),
(3) The treatment period can be set longer by selecting another drug having a different mechanism of action from the preventive or therapeutic drug for ATL.
(4) The therapeutic effect can be sustained by selecting another drug having a different mechanism of action from the preventive or therapeutic drug for ATL.
(5) A synergistic effect can be obtained by using a prophylactic or therapeutic drug for ATL in combination with another drug.

When the prophylactic or therapeutic agent for ATL of the present invention is administered to a patient as a pharmaceutical preparation, a DNMT inhibitor (for example, compound (I)) may be formulated alone, or other pharmaceutical agents or pharmaceutically acceptable You may mix with a carrier etc. and make into a formulation. The content ratio of the DNMT inhibitor (for example, compound (I)) in the pharmaceutical preparation is usually 0.1 to 100% (w / w). When another drug is added to the pharmaceutical preparation, the content ratio of the DNMT inhibitor (for example, compound (I)) is usually 0.1 to 99.9% (w / w).

The amount of the DNMT inhibitor of the present invention (eg, compound (I)) and the other drug 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. It may range from mg to about 2,000 mg, about 50 mg to about 1,000 mg, about 100 mg to about 500 mg, about 150 mg to about 500 mg, or about 150 mg to about 250 mg.

It is a figure which shows the combined use effect of OR-21 and DS-3201b in MT-1. It is a figure which shows the combined use effect of OR-21 and DS-3201b in MT-2.

The anti-ATL activity is shown below.
Example

Hereinafter, the present invention will be described in more detail with reference to Examples, but these do not limit the present invention.

The cell growth inhibitory effect when the ATL cell line (HTLV-1 infected cell line) was treated with decitabine and EPZ-6438 was evaluated. Regarding the ATL cell line, MT-1 strain and MT-2 strain were purchased from JCRB cell bank and used. Seed each cell into a 24-well plate at 40,000 cells / 1 ml / well in culture medium (RPMI-1640 containing 10% FBS and 1% Penn-Strep), and culture at 37 ° C, 5% CO ₂ for approx. 2 Incubated for hours. For MT-1, adjust the concentration of decitabine to 0.1, 0.2, 0.5, 1 μM and the concentration of EPZ-6438 to 0.5, 1, 2.5, 5 μM with the culture medium, and then use a 24-well plate. And incubated for 48 hours. For MT-2, adjust the concentration of decitabine to 0.05, 0.1, 0.2, and 0.5 μM and the concentration of EPZ-6438 to 0.25, 0.5, 1, and 2.5 μM with the culture medium, and then use a 24-well plate. And incubated for 48 hours. After 48 hours, re-add decitabine or EPZ-6438 to the same final concentration, incubate for an additional 48 hours (96 hours in total), then transfer 100 μl of cell culture medium to a 96 well plate, and use CCK-8. After reacting with reagents (DOJINDO, CK04) according to the attached manual, the absorption at 450 nm and 620 nm (Blank) of each well was measured with a plate reader (Varioskan Flash, Thermo Fisher Scientific Co., Ltd.). Table 1 and Table 2 show the relative values of each sample-treated well when the value of the sample-free well was 100%.
(Table 1)

(Table 2)

From the above Table 1, it was found that the combined treatment of decitabine (DNMT inhibitor) and EPZ-6438 (histone methyltransferase inhibitor) induces a synergistic cell growth inhibitory effect on MT-1.
From the above Table 2, it was found that the combined treatment of decitabine and EPZ-6438 induces a synergistic cell growth inhibitory effect also on MT-2.

The MT-1 strain and MT-2 strain, which are ATL cell lines (HTLV-1 infected cell line), were purchased from JCRB cell bank. Seed each cell in a culture medium (RPMI-1640 containing 10% FBS and 1% Penn-Strep) at 0.5 x 10 ⁵ cells / 0.9 ml / well on a 24-well plate and incubate at 37 ° C, 5% CO ₂ Incubated for about 3 hours. OR-21 single agent, DS-3201b single agent, and OR-21 / DS-3201b mixed solution, which were diluted with the culture solution to 5 μM -1.6 μM, were added to a 24-well plate by 100 μl and cultured for 8 days ( Compound final concentration 500-16 nM). During this time, fresh compound was re-added every 2 days. After 4 days of culture, the cell culture medium was diluted 5-fold. Regarding the antitumor effect, 100 μl of cell culture was aliquoted into a 96 well plate, and after reacting with CCK-8 reagent (DOJINDO, CK04) according to the attached manual, plate reader (Varioskan Flash, Thermo Fisher Scientific The absorption of each well was measured at 450 nm and 620 nm (Blank). The value of each sample-treated well when the value of the well without addition of the sample was 100% was expressed as a relative value, and the IC ₅₀ value was calculated. The results are shown in FIGS. OR-21 is a compound in which R, R ′ and R ² are hydrogen atoms in the formula (I) and R ¹ is a triethylsilyl group, and the name of OR-21 is 5′-O-Triethylsilyl-2. It is'-deoxy-5-azacytidine.
The morphological changes were observed with a phase contrast microscope 4 days after the compound treatment. The result is shown in FIG. From FIG. 1, the combined treatment of OR-21 (DNMT inhibitor) and DS-3201b (histone methyltransferase inhibitor) for 6 days synergistically suppresses MT-1 cell proliferation (IC ₅₀ ; OR-21. : 58 nM, DS-3201b:> 500 nM, combined treatment: 36 nM). From FIG. 2, the combined treatment of OR-21 and DS-3201b for 4 days synergistically suppresses the cell growth of MT-2 (IC ₅₀ ; OR-21: 172 nM, DS-3201b:> 500 nM, combination treatment). Treatment 118 nM), and cells adhered to and spread on the dish bottom, and it was found that the combined treatment characteristically promotes cell adhesion.

According to the present invention, a DNMT inhibitor can be combined with another drug and provided to the medical field as a preventive or therapeutic drug for cancer.

Claims

A pharmaceutical composition comprising a DNMT inhibitor in combination with other agents.
A pharmaceutical composition comprising another drug for use in combination with a DNMT inhibitor.
A pharmaceutical composition comprising a DNMT inhibitor and other agents.
The DNMT inhibitor has the formula (I):

(In the formula, R and R ′ are each an OR 3 group, a hydrogen atom, a halogen atom or an alkyl group, and R 1 , R 2 and R 3 are each a hydrogen atom or a formula (II):

(In the formula, R 4 , R 5 and R 6 are each an alkyl group, an aryl group or an arylalkyl group which may have a substituent). 4. A compound represented by the formula (1) or a salt thereof, decitabine, azacitidine, RG108, thioguanine, Zebularine, SGI-110, CC-486, SGI-1027, lomeguatrib or procainamide hydrochloride. The pharmaceutical composition according to the item.
The pharmaceutical composition according to claim 4, wherein R 1 is a silyl group represented by formula (II), and R 2 and R or R 3 are hydrogen atoms.
The pharmaceutical composition according to claim 4, wherein R 2 is a silyl group represented by formula (II), and R 1 and R or R 3 are hydrogen atoms.
The pharmaceutical composition according to claim 4, wherein R 1 and R 2 are each a silyl group represented by the formula (II), and R or R 3 is a hydrogen atom.
The pharmaceutical composition according to claim 4, wherein R 1 is a hydrogen atom, and R 2 and R 3 are each a silyl group represented by the formula (II).
The pharmaceutical composition according to claim 4, wherein each of R 1 , R 2 and R 3 is a silyl group represented by the formula (II).
The pharmaceutical composition according to claim 4, wherein each of R 1 , R 2 and R 3 is a hydrogen atom.
The R 4 , R 5 and R 6 are each a C 1 -C 8 alkyl group optionally having a substituent, a C 6 -C 10 aryl group or a C 7 -C 14 arylalkyl group. 4. The pharmaceutical composition according to 4.
The pharmaceutical composition according to claim 11, wherein the C 6 -C 10 aryl group is a phenyl group or a naphthyl group.
The pharmaceutical composition according to claim 11, wherein the C 7 -C 14 arylalkyl group is a benzyl group, a phenethyl group or a naphthylmethyl group.
The pharmaceutical composition according to any one of claims 1 to 13, wherein the other drug is an enzyme inhibitor.
The enzyme inhibitor is selected from the group consisting of histone acetylase inhibitors, histone deacetylase inhibitors, histone methylase inhibitors, histone demethylase inhibitors and ribonucleoside diphosphate reductase inhibitors The pharmaceutical composition according to claim 14, which is one or more kinds.
The pharmaceutical composition according to claim 14, wherein the enzyme inhibitor is one or more selected from the group consisting of DS-3201b, HBI-8000, trichostatin A (TSA), suramin (Suramin), EPZ005687 and Adox. ..
The pharmaceutical composition according to any one of claims 1 to 16, which further comprises a pharmaceutically acceptable carrier.
The pharmaceutical composition according to any one of claims 1, 2 and 4 to 17, wherein the DNMT inhibitor and the other drug are administered at the same time.
The pharmaceutical composition according to any one of claims 1, 2 and 4 to 17, wherein the DNMT inhibitor and the other drug are administered separately.
The pharmaceutical composition according to any one of claims 1, 2 and 4 to 17, wherein the DNMT inhibitor is administered before administration of the other drug.
The pharmaceutical composition according to any one of claims 1, 2 and 4 to 17, wherein the DNMT inhibitor is administered after administration of another drug.
The pharmaceutical composition according to any one of claims 1 to 21, for the prevention or treatment of cancer.
The pharmaceutical composition according to claim 22, wherein the cancer is leukemia.
The pharmaceutical composition according to claim 23, wherein the leukemia is ATL.
A preventive or therapeutic agent for cancer, comprising the pharmaceutical composition according to any one of claims 1 to 21.
The preventive or therapeutic drug for cancer according to claim 25, wherein the cancer is leukemia.
The preventive or therapeutic agent for cancer according to claim 26, wherein the leukemia is ATL.
A kit for preventing or treating cancer, which comprises the DNMT inhibitor according to any one of claims 1 to 22 and other drugs.
A method for preventing or treating cancer in a mammal, which comprises administering an effective amount of a DNMT inhibitor and an effective amount of another drug to the mammal.