JP2007055940A - Pyrazolopyrimidine derivative - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compound having excellent function regulatory action of CCR4, or TARC and/or MDC, and usable for prevention and treatment of several kinds of inflammatory diseases, allergic diseases, autoimmune diseases and the like. <P>SOLUTION: The compound is a new pyrazolopyrimidine derivative represented by formula (I) or a salt thereof, wherein, -R<SP>1</SP>and -R<SP>2</SP>are a 1-6C alkyl, which may be substituted with one or more halogen, or -H; -A- is single bond or methylene; -R<SP>3</SP>is a (substituted) phenyl; and -R<SP>4</SP>is a (substituted) cyclic amino. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a novel pyrazolopyrimidine derivative and a pharmaceutical comprising the same as an active ingredient, particularly a therapeutic agent for inflammatory diseases.

Chemokines, which are cell migration factors, are roughly classified into two types, CXC / α chemokines and CC / β chemokines, due to structural features. These chemokine receptors belong to the seven-transmembrane G protein-coupled receptor family, and are composed of CXC chemokine receptors and CC chemokine receptors (Non-patent Document 1).
CC chemokine receptor 4 (CCR4) has been cloned from T lymphocytes and thymus (Non-patent document 2, Non-patent document 3), and was reported to be expressed mainly in T cells initially referred to as Th2 type (Non-patent document 2). Patent Document 4). However, subsequent detailed analysis showed that CCR4 is widely present in Th1 and Th2 effector memory T cells (Non-patent Documents 5 and 6). Furthermore, recent studies have revealed that CCR4 is present in almost all skin-oriented T cells (Non-patent Document 7), monocytes / macrophages, dendritic cells, and NK cells (Non-patent Document 8). ).
CC chemokines Thymus and activation-regulated chemokine (TARC) and Macrophage-derived chemokine (MDC) are specific ligands for CCR4 (Non-patent Documents 9 and 10). TARC was found as a T cell migration factor (Non-Patent Document 11), and MDC was found as a migration factor for monocytes, macrophages and NK cells (Non-Patent Document 12). Both chemokines are characteristic of inflammatory and homeostatic chemokines. (Non-Patent Document 13).

  Many reports suggest that CCR4 and its ligands TARC and MDC are involved in various diseases such as inflammatory diseases, allergic diseases, and autoimmune diseases. For example, asthma (non-patent document 14), atopic dermatitis (non-patent document 15), psoriasis (non-patent document 16), rheumatoid arthritis (non-patent document 17), inflammatory bowel disease (non-patent document 18), etc. Can be mentioned. Therefore, CCR4 function regulators are expected as preventive or therapeutic agents for these diseases. Various drugs such as steroids are used as preventive or therapeutic agents for the above-mentioned inflammatory diseases, allergic diseases, autoimmune diseases, etc., but drugs based on a new mechanism of action in terms of their therapeutic effects and side effects The development of is eagerly desired.

［式中の記号は当該公報を参照のこと。］ For example, it has been reported that a compound represented by the following formula has a function regulating action of TARC or MDC (Patent Document 1).

[See the official gazette for symbols in the formula. ]

［式中の記号は当該公報を参照のこと。］ For example, it has been reported that a compound represented by the following formula has a CCR4 function-regulating action (Patent Document 2).

[See the official gazette for symbols in the formula. ]

［式中の記号は当該公報を参照のこと。］ In addition, a quinazoline derivative represented by the following formula has been reported which has IgE antagonistic activity and is useful as a therapeutic agent for allergic diseases and cartilage disorders (Patent Document 3).

[See the official gazette for symbols in the formula. ]

  However, there is no disclosure or suggestion about the pyrazolopyrimidine derivative according to the present invention in any of the above documents.

［式中の記号は当該公報を参照のこと。］
しかし、当該文献には、これらの化合物がホスホジエステラーゼ５阻害剤であり、ホスホジエステラーゼ５関連疾患、特に高血圧の治療等に有用である旨が記載されているが、本発明に係るCCR4の機能調節作用、及びその作用に基づく疾患の治療可能性等については言及されていないし、示唆もない。 On the other hand, compounds having the same skeleton as the pyrazolopyrimidine derivative according to the present invention have been reported (Patent Document 4, Patent Document 5, and Patent Document 6).

[See the official gazette for symbols in the formula. ]
However, this document describes that these compounds are phosphodiesterase 5 inhibitors and are useful for treatment of phosphodiesterase 5-related diseases, particularly hypertension, etc., but CCR4 function-regulating action according to the present invention, There is no mention or suggestion about the possibility of treatment of diseases based on its action.

Pharmacological Reviews, 52, 145, 2000 Biochemical and Biophysical Research Communications, 218, 337, 1996 European Journal of Immunology, 26, 3021, 1996 Journal of Experimental Medicine, 187, 875, 1998 Journal of Immunology, 166, 103, 2001 The Journal of Clinical Investigation, 108, 1331, 2001 Nature, 400, 776, 1999 Arthritis & Rheumatism, 44, 1022, 2001 Journal of Biological Chemistry, 272, 15036, 1997 Journal of Biological Chemistry, 273, 1764, 1998 Journal of Biological Chemistry, 271, 21514, 1996 Journal of Experimental Medicine, 185, 1595, 1997 Immunology Today, 20, 254, 1999 The Journal of Clinical Investigation, 107, 1357, 2001 Journal of Investigative Dermatology, 115, 640, 2000 Laboratory Investigation, 81, 335, 2001 Arthritis & Rheumatism, 44, 2750, 2001 Clinical & Experimental Immunology, 132, 332, 2003 International Publication No. WO 03/104230 Pamphlet US Patent Application Publication No. 2004/0048865 Japanese Unexamined Patent Publication No. 2000-281660 International Publication No. WO 2004/96810 Pamphlet International Publication No. WO 2005/49616 Pamphlet International Publication No. WO 2005/49617 Pamphlet

  The present inventors provide a pharmaceutical composition useful for the prevention and / or treatment of inflammatory diseases, allergic diseases, autoimmune diseases and the like based on the function-regulating action of CCR4, and further a medicament containing them. The research was conducted for the purpose of providing.

As a result of intensive studies on a compound having a CCR4 function-regulating action, the present inventors have found that a novel pyrazolopyrimidine derivative is useful as a CCR4 function-regulating agent, and completed the present invention.
That is, according to the present invention, a novel pyrazolopyrimidine derivative represented by the following formula (I) or a pharmaceutically acceptable salt thereof is provided.

［式中の記号は、以下の意味を示す。
-R¹：当該ピラゾール環の窒素上の置換基であって、１つ以上のハロゲンで置換されていてもよいC₁-C₆アルキル、若しくは-H。
-R²：１つ以上のハロゲンで置換されていてもよいC₁-C₆アルキル、若しくは-H。
-A-：単結合、若しくはメチレン。
-R³：置換されていてもよいフェニル。
-R⁴：置換されていてもよい環状アミノ。］

[The symbols in the formula have the following meanings.
-R ¹ : a substituent on the nitrogen of the pyrazole ring, C ₁ -C ₆ alkyl optionally substituted with one or more halogens, or -H.
-R ² : C ₁ -C ₆ alkyl optionally substituted with one or more halogens, or -H.
-A-: Single bond or methylene.
-R ³ : phenyl which may be substituted.
-R ⁴ : Cyclic amino optionally substituted. ]

In the formula (I), -R ¹ or -R ² is preferably methyl, ethyl, n-propyl, t-butyl, or 2,2,2-trifluoroethyl; more preferably methyl, ethyl And n-propyl; particularly preferred are methyl and ethyl. Here, -R ¹ is preferably substituted with nitrogen at the 1-position of the pyrazole ring.
Further, -R ³ in formula (I) is preferably phenyl substituted with at least one halogen; more preferably phenyl substituted with two halogens; particularly preferably 4-chloro-2 -Fluorophenyl.
In addition, -R ⁴ in the formula (I) is preferably a substituted piperidine and a substituted piperazine.

  The compound of the present invention represented by the formula (I) has a pyrazolopyrimidine skeleton and is substituted with an aniline which may be substituted at the 7-position of the pyrazolopyrimidine ring, or an optionally substituted benzylamine, It has chemical structural characteristics in that the 5-position is substituted with cyclic amino, and has pharmacological characteristics in that it has a function-regulating action of CCR4.

  Since the pyrazolopyrimidine derivative of the present invention has a function-regulating action of CCR4 or TARC and / or MDC, it is useful as a preventive and / or therapeutic agent for various inflammatory diseases, allergic diseases, autoimmune diseases and the like. Specific diseases include asthma, allergic rhinitis, allergic conjunctivitis, hay fever, dermatitis (atopic dermatitis, contact dermatitis), psoriasis, rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, insulin dependence Examples include type 1 diabetes (type 1 diabetes), rejection at the time of organ transplantation, cancer, inflammatory bowel disease (ulcerative colitis, Crohn's disease), interstitial cystitis, sepsis, pain and the like. In particular, it is expected as a preventive and / or therapeutic agent for asthma, atopic dermatitis, or rheumatoid arthritis.

Hereinafter, the present invention will be described in detail.
In the present specification, “alkyl” means a monovalent group of a linear or branched carbon chain. Therefore, “C ₁ -C ₆ alkyl” specifically includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, neopentyl, n-hexyl and the like. Preferred are C ₁ -C ₃ alkyl, methyl, ethyl, n-propyl, isopropyl.
“Halogen” is fluoro, chloro, bromo or iodo, preferably fluoro, chloro or bromo.
“Cyclic amino” has at least one nitrogen atom, and may further have one or more heteroatoms selected from the group consisting of nitrogen, oxygen and optionally oxidized sulfur. Means a monovalent group of a non-aromatic cyclic amine having 3 to 8 ring members. In particular, in -R ⁴ , the nitrogen atom of the cyclic amine does not necessarily have a bond for binding to the pyrazolopyrimidine ring, and the carbon atom of the cyclic amine and the pyrazolopyrimidine The ring may be directly bonded. Specific examples include monovalent groups such as azetidine, pyrrolidine, piperidine, azepan, azocan, piperazine, homopiperazine, morpholine, oxazepan, thiomorpholine, and thiazepan. These rings may have an unsaturated bond in a part of the ring such as dihydropyrrole, tetrahydropyridine, tetrahydroazepine, imidazolidine, oxazolidine, dihydrooxazine, thiazolidine, dihydrothiazine and the like.

  In the present specification, the permissible substituents of the words “which may be substituted” and “substituted” may be any substituents which are usually used as a substituent of each group. Two or more of these substituents may be present in each group.

Substituents allowed in “optionally substituted phenyl” of —R ³ include halogen, —OH, —OR ^Z , —OCO—R ^Z , —SH, —SR ^Z , —SO—R ^Z , _{^{-SO 2 -R Z, -SO 3 H}} , 1 or 2 sulfamoyl optionally substituted with R ^Z, 1 or 2 amino optionally substituted with R ^Z, -NHCO-R ^Z, -NHCO ₂ -R ^Z , -NHSO ₂ -R ^Z , nitro, -CHO, -CO-R ^Z , carboxyl, -CO ₂ -R ^Z , carbamoyl optionally substituted by one or two R ^Z , Mention may be made of cyano and R ^Z. Here, “R ^Z ” means —OH, —O— (C ₁ -C ₆ alkyl), —OCO— (C ₁ -C ₆ alkyl), carboxyl, —CO ₂ — (C ₁ -C ₆ alkyl) ), one or two C ₁ -C ₆ carbamoyl be alkyl substituted, cyano, one or two C ₁ -C ₆ alkyl amino optionally substituted, and from the group consisting of halogen C ₁ -C ₆ alkyl optionally substituted with one or more selected groups is shown (the same applies hereinafter).

［式中の記号は、以下の意味を示す。
-Y-：単結合、-CH₂-、-CO-、-CO-(C₁-C₆アルキレン)、-NHCO-、-N(R^Z)-CO-、-NH-、-N(R^Z)-、-CH₂NH-、若しくは-CH₂N(R^Z)-。
-R⁴⁰：C₃-C₈シクロアルキル、若しくは非芳香族ヘテロ環。なお、これらの基はそれぞれ置換されていてもよい。］ Substituents allowed in “optionally substituted cyclic amino” of —R ⁴ include halogen, —OH, —OR ^Z , —OCO—R ^Z , oxo (═O), —SH, —SR ^Z , —SO—R ^Z , —SO ₂ —R ^Z , —SO ₃ H, sulfamoyl optionally substituted with one or two R ^Z , amino optionally substituted with one or two R ^Z , -NHCO-R ^Z , -NHCO ₂ -R ^Z , -NHSO ₂ -R ^Z , nitro, -CHO, -CO-R ^Z , carboxyl, -CO ₂ -R ^Z , substituted with one or two R ^Z Mention may be made of carbamoyl, cyano and R ^Z which may have been prepared. In addition to these, examples of the substituent allowed in the “optionally substituted cyclic amino” of —R ⁴ include groups represented by the following formula (II).

[The symbols in the formula have the following meanings.
-Y-: Single bond, -CH _2- , -CO-, -CO- (C ₁ -C ₆ alkylene), -NHCO-, -N (R ^Z ) -CO-, -NH-, -N (R _{^{Z) -, - CH 2 NH-}} , or ^{_{-CH 2 N (R Z) -}} .
-R ⁴⁰ : C ₃ -C ₈ cycloalkyl, or non-aromatic heterocycle. Each of these groups may be substituted. ]

Here, “alkylene” means a linear or branched carbon chain divalent group. Accordingly, the “C ₁ -C ₆ alkylene” specifically includes, for example, methylene, ethylene, trimethylene, methylmethylene, dimethylmethylene, methylethylene, dimethylethylene, ethylmethylene, etc., preferably methylene and ethylene It is.
“Cycloalkyl” means a non-aromatic monovalent group consisting of carbon, which may partially have an unsaturated bond. Therefore, specific examples of “C ₃ -C ₈ cycloalkyl” include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclobutenyl, cyclohexenyl, cyclooctadienyl, and the like.
The “non-aromatic heterocycle” is a non-aromatic ring having 4 to 7 ring members containing one or more heteroatoms selected from the group consisting of nitrogen, oxygen, and optionally oxidized sulfur. And these may partially have an unsaturated bond. Specific examples include monovalent groups such as tetrahydrofuran, tetrahydropyran, tetrahydrothiofuran, tetrahydrothiopyran, dioxolane, and dioxane in addition to the above cyclic amino.

As the substituents allowed for “C ₃ -C ₈ cycloalkyl” which may be substituted in —R ⁴⁰ and “non-aromatic heterocycle”, halogen, —OH, —OR ^Z , —OCO— R ^Z , oxo (═O), —SH, —SR ^Z , —SO—R ^Z , —SO ₂ —R ^Z , —SO ₃ H, sulfamoyl optionally substituted by one or two R ^Z , Amino, —NHCO—R ^Z , —NHCO ₂ —R ^Z , —NHSO ₂ —R ^Z , Nitro, —CHO, —CO—R ^Z , carboxyl, —, optionally substituted with one or two R ^Z Mention may be made of CO ₂ -R ^Z , carbamoyl, cyano and R ^Z optionally substituted by one or two R ^Z.

The compound (I) of the present invention may have a geometric isomer or a tautomer depending on the type of the substituent. Moreover, this invention compound (I) may have an asymmetric carbon atom. The present invention includes separated or mixed isomers. Further, a label, that is, a compound in which one or more atoms of the compound of the present invention are substituted with a radioactive isotope or a non-radioactive isotope is also encompassed in the present invention.
Furthermore, the present invention includes so-called pharmacologically acceptable prodrugs of the compound (I) of the present invention. A pharmacologically acceptable prodrug is a compound having a group that can be converted to an amino group, a hydroxyl group, a carboxyl group, or the like of the present invention by solvolysis or under physiological conditions. As the group to be formed, the group described in Prog. Med., Volume 5, pages 2157-2161, 1985, and “Development of drugs” (Yodogawa Shoten, 1990), Volume 7, Molecular design pages 163-198 Can be mentioned.

Compound (I) may form a salt with a base depending on the type of acid addition salt or substituent. Such a salt may be a pharmaceutically acceptable salt, and specifically includes inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propion. Acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, acid addition salts with organic acids such as aspartic acid, glutamic acid, sodium And salts with inorganic bases such as potassium, magnesium, calcium and aluminum, and organic bases such as methylamine, ethylamine, ethanolamine, lysine and ornithine, and ammonium salts.
Furthermore, the present invention includes various hydrates and solvates of the compound (I) of the present invention and salts thereof, and polymorphic substances.

(Production method)
The compound (I), which is an active ingredient of the present invention, and a pharmaceutically acceptable salt thereof are produced by applying various known synthetic methods utilizing characteristics based on the basic skeleton or the type of substituent. be able to. In this case, depending on the type of functional group, it is effective in terms of production technology to protect the functional group with an appropriate protective group at the raw material or intermediate stage, or to replace it with a group that can be easily converted to the functional group. There is a case. Examples of such a functional group include an amino group, a hydroxyl group, a carboxyl group, and the like, and examples of protective groups thereof include, for example, “Protective Groups in Organic Synthesis (3rd edition, by TWGreene) and Utz (PGMWuts)”. 1999) ”, and may be appropriately selected depending on the reaction conditions. According to such a method, after carrying out the reaction by introducing the protecting group, the desired compound can be obtained by removing the protecting group as necessary or converting it to a desired group.
Moreover, the prodrug of this invention compound (I) can be manufactured by reacting using the compound (I) which introduce | transduced or obtained the specific group in the step of a raw material thru | or an intermediate body like the said protecting group. The reaction can be carried out by applying a method known by those skilled in the art, such as ordinary esterification, amidation, dehydration and the like.

（式中、-R¹、-R²、-A-、及び-R³はそれぞれ前述の意味を示す。以下同様。）
本製法は、(1a)で示されるジクロロピリミジンに対し、(1b)で示されるアニリン若しくはベンジルアミン誘導体を作用させ、式(1c)で示されるクロロピリミジン誘導体を製造する方法である。
(1a)と(1b)との反応は、常圧又は加圧下に、溶媒の不存在下若しくは適当な溶媒中で行われる。
溶媒の具体例としては、トルエン、キシレン等の芳香族炭化水素類；アセトン、メチルエチルケトン等のケトン類；エーテル、テトラヒドロフラン（THF）、ジオキサン、ジエトキシエタン等のエーテル類；メタノール（MeOH）、エタノール（EtOH）、2-プロパノール（iPrOH）等のアルコール類；アセトニトリル；ジメチルホルムアミド（DMF）、1,3-ジメチル-2-イミダゾリジノン、ジメチルスルホキシド（DMSO）等の非プロトン性極性溶媒；水；あるいはこれらの混合溶媒が挙げられる。本反応は塩基の存在下に行うのが好ましく、塩基の具体例としては、炭酸ナトリウム、炭酸カリウム等の炭酸アルカリ；炭酸水素ナトリウム、炭酸水素カリウム等の炭酸水素アルカリ；ナトリウムメトキシド、ナトリウムエトキシド、カリウムt-ブトキシド等のアルコキシド；トリエチルアミン、ジイソプロピルエチルアミン等の３級アミン；1,8-ジアザビシクロ[5.4.0]ウンデカ-7-エン、ピリジン、ルチジン等の有機塩基等を挙げることができるが、(1b)の過剰量で兼ねることもできる。反応温度は、原料化合物の種類、反応条件等により異なるが、通常室温から溶媒の還流温度程度で行うことができる。
なお、製造原料である(1b)は、その塩として反応に供することもできる。また、製造原料である(1a)は、本明細書の参考例に記載の方法、又は例えば前述の特許文献４、５、若しくは６に記載の方法、あるいはそれらに準じた方法により、容易に製造することができる。 <Intermediate production method>

(In the formula, -R ¹ , -R ² , -A-, and -R ³ each have the above-mentioned meaning. The same shall apply hereinafter.)
This production method is a method for producing a chloropyrimidine derivative represented by the formula (1c) by allowing an aniline or benzylamine derivative represented by (1b) to act on the dichloropyrimidine represented by (1a).
The reaction between (1a) and (1b) is carried out at normal pressure or under pressure, in the absence of a solvent or in a suitable solvent.
Specific examples of the solvent include aromatic hydrocarbons such as toluene and xylene; ketones such as acetone and methyl ethyl ketone; ethers such as ether, tetrahydrofuran (THF), dioxane and diethoxyethane; methanol (MeOH), ethanol ( EtOH), alcohols such as 2-propanol (iPrOH); acetonitrile; aprotic polar solvents such as dimethylformamide (DMF), 1,3-dimethyl-2-imidazolidinone, dimethyl sulfoxide (DMSO); water; or These mixed solvents are mentioned. This reaction is preferably carried out in the presence of a base. Specific examples of the base include alkali carbonates such as sodium carbonate and potassium carbonate; alkali hydrogen carbonates such as sodium bicarbonate and potassium bicarbonate; sodium methoxide and sodium ethoxide. Alkoxides such as potassium t-butoxide; tertiary amines such as triethylamine and diisopropylethylamine; and organic bases such as 1,8-diazabicyclo [5.4.0] undec-7-ene, pyridine and lutidine. An excess amount of (1b) can also be used. While the reaction temperature varies depending on the type of raw material compound, reaction conditions, and the like, it can be usually carried out at room temperature to the reflux temperature of the solvent.
The production raw material (1b) can also be subjected to the reaction as a salt thereof. The production raw material (1a) can be easily produced by the method described in the Reference Examples of the present specification, or the method described in Patent Document 4, 5, or 6 described above, or a method based thereon. can do.

（式中、-R⁴は前述の意味を示す。以下同様。）
本製法は、中間体製法で製造した(1c)に対し、(2a)で示される環状アミン誘導体を作用させ、式（Ｉ）で示される本発明化合物を製造する方法である。
反応は、中間体製法で示した方法に準じて行うことができる。また、本明細書の実施例に記載の方法、若しくはそれに準じた方法を採用することができる。 <First manufacturing method>

(In the formula, -R ⁴ has the above-mentioned meaning. The same applies hereinafter.)
This production method is a method for producing the compound of the present invention represented by the formula (I) by reacting (1c) produced by the intermediate production method with the cyclic amine derivative represented by (2a).
The reaction can be carried out according to the method shown in the intermediate production method. Moreover, the method as described in the Example of this specification, or the method according to it can be employ | adopted.

（式中、-R⁴¹は、前述の式（ＩＩ）等の、-R⁴の「置換されていてもよい環状アミノ」において許容される置換基を示す。）
本製法は、中間体製法で製造した(1c)に対し、(3a)で示されるピペラジン若しくはその保護体を作用させ、必要に応じた脱保護により得られるピペラジン誘導体(3b)に対して適切な官能基導入を行うことにより、式（Ｉ）で示される本発明化合物のうち、-R⁴が置換されていてもよい環状アミノのうちの置換されていてもよいピペラジンである場合の式（Ｉ’）で示される本発明化合物を製造する方法である。
step 3-1は、(1c)のクロロをピペラジンで置換する反応であり、(3a)として使用するピペラジンは、保護されていないピペラジンはもちろん、例えばt-ブチルオキシカルボニル、ベンジルオキシカルボニル、メトキシカルボニル、エトキシカルボニル等のアミノ基の保護基で一方の窒素原子が保護されたピペラジンを用いることができる。また、これらのピペラジン誘導体は、塩を形成した状態で反応に供することもできる。
反応は、中間体製法で示した方法に準じて行うことができる。また、本明細書の参考例若しくは実施例に記載の方法、又はそれに準じた方法を採用することができる。また、(3a)として保護されたピペラジンを用いた場合には、その保護基を脱保護するに適した反応により脱保護することにより(3b)に導くことができる。これらの脱保護反応については、前述の「Protective Groups in Organic Synthesis（第3版、1999年）」に記載の方法を参照することができる。
step 3-2は、(3b)のピペラジン上の窒素原子に官能基を導入する反応であり、代表的には、アシル化、アルキル化を挙げることができる。
例えば、アシル化は、当業者が通常用いるアシル化反応を採用することができるが、特に1-ヒドロキシベンゾトリアゾール（HOBt）存在下、1-エチル-3-(3-ジメチルアミノプロピル)カルボジイミド塩酸塩（EDCI）や、ジシクロヘキシルカルボジイミド、カルボニルジイミダゾール、ジフェニルホスホリルアジド、ジエチルホスホリルシアニド等の縮合剤を作用させる方法が好適に用いられる。使用する縮合剤等の反応条件により異なるが、通常、エーテル類；芳香族炭化水素類；ジクロロメタン、ジクロロエタン、クロロホルム等のハロゲン化炭化水素類；酢酸エチル（EtOAc）等エステル類；アセトニトリル；非プロトン性極性溶媒等の反応に不活性な有機溶媒中、冷却下、冷却乃至室温下、若しくは室温乃至加熱下に行われる。
また、例えばアルキル化は、当業者が通常用いるアルキル化反応を採用することができ、エーテル類；芳香族炭化水素類；ジクロロメタン、ジクロロエタン、クロロホルム等のハロゲン化炭化水素類；アセトニトリル；非プロトン性極性溶媒等の反応に不活性な有機溶媒中、冷却下、冷却乃至室温下、若しくは室温乃至加熱下に、炭酸アルカリ；炭酸水素アルカリ；アルコキシド；３級アミン；有機塩基等の塩基存在下に行うことができる。 <Second manufacturing method>

(In the formula, —R ⁴¹ represents a substituent permitted in “optionally substituted cyclic amino” of —R ⁴ such as the aforementioned formula (II)).
This production method is suitable for the piperazine derivative (3b) obtained by deprotection as necessary by reacting the piperazine represented by (3a) or its protector with (1c) produced by the intermediate production method. By introducing a functional group, among the compounds of the present invention represented by the formula (I), -R ⁴ is an optionally substituted piperazine among the optionally substituted cyclic aminos (I This is a method for producing the compound of the present invention represented by ').
Step 3-1 is a reaction of substituting chloro in (1c) with piperazine, and the piperazine used as (3a) is not only unprotected piperazine but also, for example, t-butyloxycarbonyl, benzyloxycarbonyl, methoxycarbonyl Piperazine in which one nitrogen atom is protected with an amino-protecting group such as ethoxycarbonyl can be used. Moreover, these piperazine derivatives can also be subjected to the reaction in the form of a salt.
The reaction can be carried out according to the method shown in the intermediate production method. Moreover, the method as described in the reference example or Example of this specification, or the method according to it can be employ | adopted. In addition, when a protected piperazine is used as (3a), it can be led to (3b) by deprotecting the protecting group by a reaction suitable for deprotection. For these deprotection reactions, the method described in “Protective Groups in Organic Synthesis (3rd edition, 1999)” can be referred to.
Step 3-2 is a reaction for introducing a functional group into the nitrogen atom on piperazine in (3b), and representative examples include acylation and alkylation.
For example, acylation can employ an acylation reaction commonly used by those skilled in the art, and in particular, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride in the presence of 1-hydroxybenzotriazole (HOBt). A method in which a condensing agent such as (EDCI), dicyclohexylcarbodiimide, carbonyldiimidazole, diphenylphosphoryl azide, or diethylphosphoryl cyanide is used is preferably used. Usually, ethers; aromatic hydrocarbons; halogenated hydrocarbons such as dichloromethane, dichloroethane, and chloroform; esters such as ethyl acetate (EtOAc); acetonitrile; aprotic, depending on the reaction conditions such as the condensing agent used The reaction is carried out in an organic solvent inert to the reaction, such as a polar solvent, under cooling, from cooling to room temperature, or from room temperature to heating.
In addition, for example, alkylation can be performed by an alkylation reaction commonly used by those skilled in the art. Ethers; aromatic hydrocarbons; halogenated hydrocarbons such as dichloromethane, dichloroethane and chloroform; acetonitrile; aprotic polarity In an organic solvent inert to the reaction, such as a solvent, under cooling, from cooling to room temperature, or from room temperature to heating, in the presence of a base such as alkali carbonate; alkali hydrogen carbonate; alkoxide; tertiary amine; organic base. Can do.

  Furthermore, some of the compounds represented by the formula (I) can be obtained from the compound of the present invention produced as described above by known alkylation, acylation, substitution reaction, oxidation, reduction, hydrolysis, deprotection, etc. It can also be produced by arbitrarily combining processes that can be usually employed by a trader.

The reaction product obtained by each of the above production methods is isolated and purified as various solvates such as a free compound, a salt thereof or a hydrate. The salt can be produced by subjecting it to normal salt formation treatment.
Isolation and purification are performed by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various chromatography.
Various isomers can be isolated by a conventional method utilizing the difference in physicochemical properties between isomers. For example, optical isomers can be separated by a general optical resolution method such as fractional crystallization or chromatography. Optical isomers can also be produced from appropriate optically active raw material compounds.

  The pharmacological activity of the compound of the present invention was confirmed by the following test.

1. Action on [ ³⁵ S] GTPγS binding test via CCR4 (1) Acquisition of human CCR4-expressing cell line
A vector (including a neomycin resistance gene) in which the human CCR4 gene was inserted downstream of the EF-1α promoter was prepared, and transfected into the mouse pre B cell line B300-19 cells by electroporation. These cells were cultured in a G418-added medium, and a single cell line that constantly and stably expresses human CCR4 was obtained by the limiting dilution method.
(2) Preparation of human CCR4-expressing cell line membrane fraction Human CCR4-expressing cells were collected and washed with PBS, and then suspended in Lysis Buffer (10 mM Hepes pH 7.5, 2 mM EDTA, protainase inhibitor). The suspension was placed on ice for 15 minutes, and then the cells were disrupted by a homogenizer and centrifuged (20000 rpm, 10 min, 4 ° C.). The supernatant was further ultracentrifuged (22K, 30 min, 4 ° C.), and the pellet was suspended in PBS and used as a membrane fraction in subsequent experiments.
(3) GTPγS binding test Each concentration of the test compounds was 20 mM Hepes pH 7.05, 100 mM NaCl, 5 mM MgCl ₂ , GDP 2 μM, Human MDC, [ ³⁵ S] GTPγS 150 pM, Wheatgerm agglutinin SPA beads 1 mg and Radioactivity was measured by reacting at room temperature for 1 hour 30 minutes in a reaction mixture containing 1 μg of human CCR4-expressing cell line membrane fraction.
As a result, the compounds of Example 10, Example 26, Example 33, Example 37, and Example 53 showed an inhibitory activity value (IC ₅₀ ) of 100 nM or less.

2. Action on mouse oxazolone-induced contact dermatitis
Balb / c mice (6-10 weeks old, female, Charles River Japan) were sensitized by applying 150 μl of 3% oxazolone / ethanol solution (Sigma Aldrich Japan) to the abdomen. On the 6th day after sensitization, 10 μl of 1% oxazolone / ethanol solution was applied to both sides of the right ear. Test drug administration was performed 12 hours after application of the oxazolone solution (test drug administration group), and only the solvent used to dissolve the test drug was administered to the control group. The thickness of the right auricle was measured using a thickness gauge (Mitutoyo) before and 20 hours after application, and swelling (thickness increase = measured value after 20 hours−measured value before application) was calculated. The inhibition rate was calculated according to the following formula, assuming that the group to which oxazolone solution was applied without sensitization was the normal group. In addition, the said test was implemented with 5 animals per group.
Inhibition rate = (swelling in control group−swelling in test drug administration group) × 100 / (swelling in control group−swelling in normal group)
As a result, the compound of Example 1 showed good inhibitory activity when orally administered at 30 mg / kg.

3. Action on mouse collagen-induced arthritis The action on mouse collagen-induced arthritis can be evaluated using the method described in The Japanese Journal of Pharmacology, 88, 332 (2002).
Besides the above test examples, for example, a mouse asthma model described in Immunology, 98, 345 (1999), an oxazolone-induced chronic contact dermatitis model described in Journal of Investigative Dermatorogy, 111, 86 (1998) (atopic) The pharmacological action of the compound of the present invention can be confirmed by various evaluation models generally used for evaluating anti-inflammatory action such as dermatitis model).
From the above test results, the compound of the present invention has a function-regulating action of CCR4 or TARC and / or MDC, and therefore is useful as a prophylactic / therapeutic agent for various inflammatory diseases, allergic diseases, autoimmune diseases, etc. Was confirmed.

A preparation containing one or more of compound (I) or a salt thereof as an active ingredient is usually prepared using carriers, excipients, and other additives used for formulation.
Administration is oral by tablets, pills, capsules, granules, powders, liquids, etc., or parenteral by injections such as intravenous injections, intramuscular injections, suppositories, transdermal agents, nasal agents or inhalants. Either form may be sufficient. The dosage is appropriately determined according to the individual case, taking into account the symptoms, age of the subject, sex, etc. In general, in the case of oral administration, it is about 0.001 mg / kg to 100 mg / kg per day for an adult. This is administered once, or divided into 2 to 4 times. In addition, when intravenously administered depending on symptoms, it is usually administered once or multiple times in a range of 0.0001 mg / kg to 10 mg / kg per adult. In the case of inhalation, it is usually administered once or several times a day in the range of 0.0001 mg / kg to 1 mg / kg per adult.
As the solid composition for oral administration according to the present invention, tablets, powders, granules and the like are used. In such solid compositions, one or more active substances are present in at least one inert excipient such as lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinylpyrrolidone, metasilicate. Mixed with magnesium aluminate acid. The composition may contain an inert additive, for example, a lubricant such as magnesium stearate, a disintegrant such as sodium carboxymethyl starch, and a solubilizing agent according to a conventional method. If necessary, tablets or pills may be coated with a sugar coating or a gastric or enteric coating agent.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, etc., and include commonly used inert solvents such as purified water, ethanol, etc. . In addition to the inert solvent, the composition may contain adjuvants such as solubilizers, wetting agents, and suspending agents, sweeteners, corrigents, fragrances, and preservatives.
Injections for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of the aqueous solvent include distilled water for injection and physiological saline. Non-aqueous solvents include, for example, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, alcohols such as ethanol, polysorbate 80 (Pharmacopeia name), and the like. Such a composition may further contain isotonic agents, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, and solubilizing agents. These are sterilized by, for example, filtration through a bacteria-retaining filter, blending with a bactericide or irradiation. These can also be used by producing a sterile solid composition and dissolving and suspending it in sterile water or a sterile solvent for injection before use.
Transmucosal agents such as inhalants and nasal agents are used in the form of solids, liquids, and semisolids, and can be produced according to conventionally known methods. For example, excipients such as lactose and starch, and pH adjusters, preservatives, surfactants, lubricants, stabilizers, thickeners and the like may be added as appropriate. For administration, an appropriate device for inhalation or insufflation can be used. For example, using a known device such as a metered dose inhalation device or a nebulizer, the compound is administered alone or as a powder in a formulated mixture or as a solution or suspension in combination with a pharmaceutically acceptable carrier. be able to. The dry powder inhaler or the like may be for single or multiple administration, and a dry powder or a powder-containing capsule can be used. Alternatively, it may be in the form of a pressurized aerosol spray using a suitable propellant, for example, a suitable gas such as chlorofluoroalkane, hydrofluoroalkane or carbon dioxide.
External preparations include ointments, plasters, creams, jellies, poultices, sprays, lotions, eye drops, eye ointments and the like. Contains commonly used ointment bases, lotion bases, aqueous or non-aqueous solutions, suspensions, emulsions, and the like. For example, as an ointment or lotion base, polyethylene glycol, carboxyvinyl polymer, white petrolatum, honey beeswax, polyoxyethylene hydrogenated castor oil, glyceryl monostearate, stearyl alcohol, cetyl alcohol, lauromacrogol, sorbitan sesquioleate, etc. Is mentioned.

Hereinafter, based on an Example, the manufacturing method of this invention compound (I) is demonstrated in detail. The present invention is not limited to the compound inventions described in the following examples. Moreover, the manufacturing method of a raw material compound is shown in a reference example. In addition, the following abbreviations are used in the reference examples, examples, and tables below.
F: FAB-MS (M + H) ⁺ , FN: FAB-MS (MH) ⁻ , ES: ESI-MS (M + H) ⁺ , ESN: ESI-MS (MH) ⁻ , EI: EI-MS ( M ⁺ ), AP: APCI-MS (M + H) ⁺ , APN: APCI-MS (MH) ⁻ .

Reference example 1
4-amino-1,3-dimethyl-1H-pyrazole-5-carboxamide is reacted with diphenyl carbonate in the presence of potassium carbonate to give 1,3-dimethyl-1H-pyrazolo [4,3-d] pyrimidine-5,7 (4H, 6H) -dione was obtained. This was reacted with phosphorus oxychloride in the presence of tri-n-propylamine to obtain 5,7-dichloro-1,3-dimethyl-1H-pyrazolo [4,3-d] pyrimidine.
ES: 218.

Reference example 2
4-amino-3-ethyl-1-methyl-1H-pyrazole-5-carboxamide is reacted with triphosgene in the presence of triethylamine to give 3-ethyl-1-methyl-1H-pyrazolo [4,3-d] pyrimidine-5 7 (4H, 6H) -dione was obtained. This was reacted with phosphorus oxychloride in the presence of tri-n-propylamine to obtain 5,7-dichloro-3-ethyl-1-methyl-1H-pyrazolo [4,3-d] pyrimidine.
F: 231.

The compounds of Reference Example 3 to Reference Example 10 shown in Table 1 below were produced using the corresponding raw materials in the same manner as in Reference Example 1 or Reference Example 2.
The symbols in the table have the following meanings.
Rf: Reference example number.
Data: Physical data.
R ¹ and R ² : substituents in the general formula (Me: methyl, Et: ethyl, FEt: 2,2,2-trifluoroethyl, nPr: n-propyl, tBu: tert-butyl, Ph: phenyl, Bn : Benzyl, Boc: tert-butyloxycarbonyl, TBS: tert-butyldimethylsilyl, di: di, tri: tri, the number before the substituent indicates the substitution position, for example 2,6-diF-4- Cl-Ph means 2,6-difluoro-4-chlorophenyl, 2-F-4-Cl-Bn means 2-fluoro-4-chlorobenzyl, and so on.)
Syn: Production method (numbers indicate that they were synthesized using the corresponding raw materials in the same manner as the reference example compounds having the numbers as reference example numbers).

Reference Example 11
In the same manner as in Reference Example 2, 5,7-dichloro-2-ethyl-3-methyl-2H-pyrazolo [4,3-d] pyrimidine was obtained.
F: 231.

Reference Example 11-a
In the same manner as in Reference Example 2, 5,7-dichloro-2,3-dimethyl-2H-pyrazolo [4,3-d] pyrimidine was obtained.
F: 217.

Reference Example 11-b
In the same manner as in Reference Example 2, 5,7-dichloro-3-ethyl-2-methyl-2H-pyrazolo [4,3-d] pyrimidine was obtained.
F: 231.

Reference Example 12
5,7-dichloro-1,3-dimethyl-1H-pyrazolo [4,3-d] pyrimidine is reacted with 4-chloro-2-fluoroaniline in the presence of potassium t-butoxide to give 5-chloro-N- ( 4-Chloro-2-fluorophenyl) -1,3-dimethyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine was obtained.
ES: 327.

Reference Example 17
5,7-dichloro-1-methyl-3-propyl-1H-pyrazolo [4,3-d] pyrimidine is reacted with 4-chloro-2-fluorobenzylamine hydrochloride in the presence of diisopropylethylamine to give 5-chloro- N-[(4-chloro-2-fluorophenyl) methyl] -1-methyl-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine was obtained.
ES: 369.

  The compounds of Reference Example 13 to Reference Example 16 and Reference Example 18 to 25 shown in Table 2 below were produced using the corresponding raw materials in the same manner as in Reference Example 12 or Reference Example 17.

Reference Example 26
In the same manner as in Reference Example 12, 5-chloro-N- (4-chloro-2-fluorophenyl) -2-ethyl-3-methyl-2H-pyrazolo [4,3-d] pyrimidin-7-amine was added. Obtained.
F: 340.

Reference Example 26-a
In the same manner as in Reference Example 12, 5-chloro-N- (4-chloro-2-fluorophenyl) -2,3-dimethyl-2H-pyrazolo [4,3-d] pyrimidin-7-amine was obtained. .
F: 326.

Reference Example 26-b
In the same manner as in Reference Example 12, 5-chloro-N- (4-chloro-2-fluorophenyl) -3-ethyl-2-methyl-2H-pyrazolo [4,3-d] pyrimidin-7-amine was added. Obtained.
F: 340.

Reference Example 27
By reacting 5-chloro-N-[(4-chloro-2-fluorophenyl) methyl] -1-methyl-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine with piperazine N-[(4-Chloro-2-fluorophenyl) methyl] -1-methyl-5-piperazin-1-yl-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine was obtained. It was.
ES: 418.

  The compounds of Reference Example 28 to Reference Example 31 shown in Table 3 below were produced in the same manner as in Reference Example 27 using the corresponding starting materials.

Reference Example 32
A mixture of tert-butyl 4-oxopiperidine-1-carboxylate, 3-piperidinemethanol, 10% palladium on carbon and MeOH was stirred under a hydrogen atmosphere to give tert-butyl 3- (hydroxymethyl) -1,4 ′. -Bipiperidine-1'-carboxylate was obtained. ES: 299.

Reference Example 33
tert-Butyl (3S) -3- (ethoxycarbonyl) -1,4′-bipiperidine in the same manner as in Reference Example 34 using tert-butyl 4-oxopiperidine-1-carboxylate and ethyl (3S) -nipecotate -1′-carboxylate (ES: 341) was obtained. This THF solution was added to a THF suspension of lithium boron hydroxide and stirred under heating to reflux to obtain tert-butyl (3S) -3- (hydroxymethyl) -1,4′-bipiperidine-1′-carboxylate. Got.
ES: 299.

Reference Example 35
tert-Butyl 3- (ethoxycarbonyl) -1,4'-bipiperidine-1'-carboxylate is hydrolyzed with aqueous sodium hydroxide to give 1 '-(tert-butoxycarbonyl) -1,4'-bipiperidine-3 After making -carboxylic acid, this was reacted with EDCI, HOBT, and ammonium carbonate to obtain the target compound tert-butyl 3- (aminocarbonyl) -1,4'-bipiperidine-1'-carboxylate.
F: 312.

Reference Example 37
2,4'-bipiperidine dihydrochloride is reacted with bis tert-butyl dicarbonate in the presence of triethylamine to give tert-butyl 2,4'-bipiperidine-1'-carboxylate, and then in the presence of potassium carbonate, [(3-Bromopropyl) oxy] tert-butyldimethylsilane is reacted to give the target compound tert-butyl 1 (3-{[tert-butyldimethylsilyl] oxy} propyl) -2,4′-bipiperidine-1 '-Carboxylate was obtained.
F: 441.

Reference Example 39
A mixture of tert-butyl 4-oxopiperidine-1-carboxylate, cyclobutylamine, 10% palladium on carbon and methanol is stirred under a hydrogen atmosphere to obtain tert-butyl 4- (cyclobutylamino) piperidine-1-carboxylate. It was.
ES: 255.

  The compounds of Reference Example 34, Reference Example 36, Reference Example 38, Reference Example 38-a, and Reference Example 40 shown in Table 4 below correspond to the methods of Reference Example 33, Reference Example 35, or Reference Example 37. Manufactured using raw materials.

Reference Example 41
Treat tert-butyl 3- (hydroxymethyl) -1,4'-bipiperidine-1'-carboxylate with 4M HCl-EtOAc solution in MeOH to give 1,4'-bipiperidine-3-methanol dihydrochloride. Obtained.
ES: 199.

  The compounds of Reference Example 42 to Reference Example 49 shown in Table 5 below were produced in the same manner as in Reference Example 41 using the corresponding starting materials. However, all these compounds were produced as dihydrochloride.

Example 1
5-chloro-N- (4-chloro-2-fluorophenyl) -1-methyl-3-propyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine (200 mg), 1,4'- Bipiperidin-3-ylmethanol dihydrochloride (151 mg) and DBU (258 mg) were added to 1,2-diethoxyethane (40 mL), and the mixture was heated to reflux for 4 days. After the reaction solution was concentrated, chloroform and water were added to the residue, the organic layer was separated, and the aqueous layer was further extracted with chloroform. The organic layers were combined, washed with saturated brine, dried over magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel flash column chromatography to obtain (1 ′-{7-[(4-chloro-2-fluorophenyl) amino] -1-methyl-3-propyl-1H-pyrazolo [4,3- d] pyrimidin-5-yl} -1,4′-bipiperidin-3-yl) methanol (89 mg) was obtained. This was dissolved in MeOH (60 mL) and 4M HCl-EtOAc solution (0.13 mL) was added. After the solvent was distilled off under reduced pressure, the resulting residue was washed with diethyl ether to give (1 ′-{7-[(4-chloro-2-fluorophenyl) amino] -1-methyl-3-propyl- 1H-pyrazolo [4,3-d] pyrimidin-5-yl} -1,4′-bipiperidin-3-yl) methanol dihydrochloride (85 mg) was obtained as a white powder.

  The compounds of Examples 2 to 25, Example 35, and Examples 38 to 52 shown in Tables 6 to 12 below were produced using the corresponding raw materials in the same manner as in the method of Example 1.

Example 26
DMF (10 mL) was added to N-[(4-chloro-2-fluorophenyl) methyl] -3-methyl-5-piperazin-1-yl-1H-pyrazolo [4,3-d] pyrimidin-7-amine ( 231 mg), (2R) -1- (tert-butoxycarbonyl) piperidine-2-carboxylic acid (140 mg), HOBt (91 mg), and EDCI (128 mg) were sequentially added, and the mixture was stirred at room temperature for 4 days. Chloroform and saturated aqueous sodium hydrogen carbonate solution were added to the reaction solution, and the organic layer was separated. Next, the organic layer was washed with saturated brine, dried over magnesium sulfate, filtered, and the solvent was distilled off under reduced pressure. The residue was purified by silica gel flash column chromatography to obtain tert-butyl (2R) -2-{[4- (7-{[(4-chloro-2-fluorophenyl) methyl] amino} -3-methyl- 1H-pyrazolo [4,3-d] pyrimidin-5-yl) piperazin-1-yl] carbonyl} piperidine-1-carboxylate (181 mg) was obtained. This was dissolved in MeOH (5 mL), 4M HCl-EtOAc solution (0.79 mL) was added, and the mixture was stirred at room temperature for 3 days. After the solvent was distilled off under reduced pressure, the resulting residue was washed with diisopropyl ether to give the target compound N-[(4-chloro-2-fluorophenyl) methyl] -3-methyl-5- {4- [ (2R) -Piperidin-2-ylcarbonyl] piperazin-1-yl} -1H-pyrazolo [4,3-d] pyrimidin-7-amine dihydrochloride (144 mg) was obtained as a pale yellow solid.

The compounds of Examples 27 to 34, Example 36, and Example 37 shown in Tables 6 to 12 below were produced in the same manner as in Example 27 using the corresponding raw materials.
The symbols in the table have the following meanings.
Ex: Example number.
Salt: salt (HCl: hydrochloride, 2HCl: dihydrochloride, not described: free form).
R ¹ / R ² / A / R ³ / X, R ⁴ : Substituent in the general formula (sb: single bond. Therefore, for example, in the column of “R ¹ / R ² / A / R ³ / X”, “Me / nPr / sb / 2-F-4-Cl-Ph / CH ”, the structure of the compound of the example number is as follows: R ¹ is methyl, R ² is n-propyl, A Is a single bond, R ³ is 2-fluoro-4-chlorophenyl, and X is CH. The same applies hereinafter.

Example 53
Similar to the method of Example 1, [(3S) -1 ′-{7-[(4-chloro-2-fluorophenyl) amino] -2-ethyl-3-methyl-2H-pyrazolo [4,3- d] pyrimidin-5-yl} -1,4′-bipiperidin-3-yl] methanol dihydrochloride was obtained.
F: 502.

Example 54
Similar to the method of Example 1, (1 ′-{7-[(4-chloro-2-fluorophenyl) amino] -2,3-dimethyl-2H-pyrazolo [4,3-d] pyrimidine-5- Yl} -1,4′-bipiperidin-3-yl) methanol dihydrochloride was obtained.
F: 488.

Example 55
Similar to the method of Example 1, [(3S) -1 ′-{7-[(4-chloro-2-fluorophenyl) amino] -3-ethyl-2-methyl-2H-pyrazolo [4,3- d] pyrimidin-5-yl} -1,4′-bipiperidin-3-yl] methanol dihydrochloride was obtained.
F: 502.

Table 13 below shows the NMR data for some example compounds. In addition, these data extract and describe only the characteristic peak of each compound. Moreover, the symbol in a table | surface shows the following meaning.
NMR: NMR data (δ (ppm) of a peak in ¹ H-NMR using tetramethylsilane as an internal standard and DMSO-d ₆ as a measurement solvent).

Example 101
5-chloro-N- (4-chloro-2-fluorophenyl) -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine (0.5 g), 4-piperidinol (0.45 g), 1,8-diazabicyclo [5.4.0] undec-7-ene (0.56 ml), and dioxane (20 ml) are heated and stirred at 100 ° C. to give 1- {7-[(4- Chloro-2-fluorophenyl) amino] -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl} piperidin-4-ol (0.23 g) was obtained.
A mixed solution of the above product (0.35 g), triethylamine (1.2 ml), DMSO (1.2 ml), and dichloromethane (4 ml) was prepared separately from SO ₃ • pyridine (0.55 g), DMSO (1.2 ml), dichloromethane. (4 ml) Dropped into the mixed solution with stirring under ice-cooling, stirred at 4 ° C for 2 hours, then SO ₃ • pyridine (0.29 g) was added and stirred at 4 ° C for 1 hour, and water (4 ml) was added. In addition, extraction with chloroform-iPrOH, washing with saturated brine, and crude purified 1- {7-[(4-chloro-2-fluorophenyl) amino] -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl} piperidin-4-one was obtained.
A solution prepared by adding dichloromethane (27 ml) to this crude product was dispensed into a test tube (0.9 ml), and cyclopentylamine (8.5 mg), acetic acid (0.1 ml), sodium triacetoxyborohydride (21 mg) ) Was added and stirred at room temperature for 10 hours. To this reaction solution was added 1M aqueous sodium hydroxide solution (3 ml), extracted with chloroform (4 ml), the organic layer was washed, the solvent was distilled off under reduced pressure, and the resulting residue was treated with an appropriate solvent (approximately 0.5 ml). And purified by a preparative column (Waters Symmetry column C 18 5 μm 19ψ × 100 mm) (0.1% formic acid aqueous solution / MeOH 0 min (9: 1) -1 min (9: 1) -9 min) (5:95) -12 minutes (5:95)) and N- (4-chloro-2-fluorophenyl) -5- [4- (cyclopentylamino) piperidin-1-yl] -1-ethyl-3 -Methyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine (3.8 mg) was obtained.

  The compounds of Examples 102 to 114 shown in Table 14 below were produced using the corresponding raw materials in the same manner as in the method of Example 101.

Example 115
A mixture of 1-benzylpiperidin-4-one (2 g), 40% aqueous methylamine (5.4 ml), sodium triacetoxyborohydride (2.7 g), dichloroethane (20 ml) and acetic acid (2 ml) was added at room temperature. Upon stirring, a crude product (2.2 g) of 1-benzyl-N-methylpiperidin-4-amine was obtained.
Triethylamine (3 ml) and di-tert-butyl dicarbonate (3 ml) were added to a THF (40 ml) solution of this product, and tert-butyl (1-benzylpiperidin-4-yl) methylcarbamate (2.2 ml) was added. g) was obtained.
Add 10% palladium on carbon (100 mg) to EtOH solution (10 ml) of this product and stir at room temperature under hydrogen atmosphere to obtain tert-butylmethyl (piperidin-4-yl) carbamate (1.2 g). It was.
This product (0.48 g) and 5-chloro-N- (4-chloro-2-fluorophenyl) -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-7-amine (0.5 g), a mixed solution of 1,8-diazabicyclo [5.4.0] undec-7-ene (0.67 ml) and dioxane (5 ml) is heated and stirred at 100 ° C. to give (1- {7-[( 4-chloro-2-fluorophenyl) amino] -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl} piperidin-4-yl) methylcarbamate (0.2 g) Obtained.
To this product was added trifluoroacetic acid (5 ml) and crude N- (4-chloro-2-fluorophenyl) -1-ethyl-3-methyl-5- [4- (methylamino) piperidine-1 -Il] -1H-pyrazolo [4,3-d] pyrimidin-7-amine (0.17 g) was obtained.
A mixed solution prepared by adding HOBt (55 mg) and DMF (15 ml) to this product (0.17 g) is dispensed into a test tube (0.5 ml), and further N-methyl-L-proline (13 mg) PS-carbodiimide (manufactured by Argonaut, approximately 0.1 g) was added and stirred at room temperature for 10 hours. The resin is removed with a suitable filter, and the residue obtained by distilling off the organic solvent is dissolved with a suitable solvent (approximately 0.5 ml), and a preparative column (Waters Symmetry column C 18 5 μm 19ψ × 100 mm)) (0.1% aqueous formic acid / MeOH 0 min (9: 1) -1 min (9: 1) -9 min (5:95) -12 min (5:95)) and (S) -N -(1- {7-[(4-Chloro-2-fluorophenyl) amino] -1-ethyl-3-methyl-1H-pyrazolo [4,3-d] pyrimidin-5-yl} piperidin-4-yl ) -N, 1-dimethylpyrrolidine-2-carboxamide (3.9 mg) was obtained.

The compounds of Examples 116 to 154 shown in Tables 14 to 16 below were produced using the corresponding raw materials in the same manner as in the method of Example 115. When a raw material having a secondary amine structure is used as the raw material corresponding to N-methyl-L-proline in Example 115, a carboxylic acid derivative in which the amine is protected with tert-butyloxycarbonyl is used in the above method. Then, the tert-butyloxycarbonyl was deprotected to obtain the target Example compound.
Moreover, the symbol in a table | surface shows the following meaning.
Ex: Example number.
R: a substituent in the general formula (cPen: cyclopentyl, cHex: cyclohexyl, cHep: cycloheptyl, THP: tetrahydropyranyl, pipe: piperidinyl, pyrr: pyrrolidinyl, mor: morpholinyl, for example NH- (1-CO ₂ Et-4-pipe) is 1-ethoxycarbonylpiperidin-4-ylamino, NMeCO- (4,4-diF-2 (S) -pipe) is (2S)-(4,4-difluoropiperidin-2-yl) Carbonyl) (methyl) amino, NEtCO—CH ₂ -3 (RS) -pyrr means (3RS)-(pyrrolidin-3-ylmethylcarbonyl) (ethyl) amino, and so on.)

Tables 17 to 22 below show the structures of other compounds of the present invention. These can be easily produced by using the methods described in the above Reference Examples and Examples, methods obvious to those skilled in the art, or variations thereof. The symbols in the table have the following meanings.
No: Compound number.
Str: Chemical structure.

Claims (1)

A pyrazolopyrimidine derivative represented by the formula (I) or a pharmaceutically acceptable salt thereof.

[The symbols in the formula have the following meanings.
-R ¹ : a substituent on the nitrogen of the pyrazole ring, C ₁ -C ₆ alkyl optionally substituted with one or more halogens, or -H.
-R ² : C ₁ -C ₆ alkyl optionally substituted with one or more halogens, or -H.
-A-: Single bond or methylene.
-R ³ : phenyl which may be substituted.
-R ⁴ : Cyclic amino optionally substituted. ]