JP2004528269A - Use of vitamin D derivatives as bone resorption inhibitors - Google Patents

Abstract

The present invention provides a novel bone resorption inhibitor.
A bone resorption inhibitor comprising a vitamin D antagonist as an active ingredient.

Description

【００１０】
［式 （１） において、ｍは１から３の整数を表し、ｑは０から３の整数を表し、ｒは０から３の整数を表し、Ｘは炭素原子または酸素原子表す。ただし、１ ≦ ｑ ＋ ｒ ≦ ３である。］
により表される化合物があげられる。
【００１１】
この中で、ｍが１または２である化合物は好ましい。さらに、ｍ、ｑ、ｒおよびＸの組み合わせに関すると、表１に示す化合物は好ましく、この中で特に化合物Ｎｏ． １１、１３、１６、２１、２３および２６は好ましい。表１に示す化合物において、構造中に不斉炭素原子が存在する場合、それは（Ｓ） 配置および（Ｒ） 配置の両方を含む。
【００１２】
【表１】

【００１３】
さらに、本発明のビタミンＤアンタゴニストのサンプルとしては、下記式 （２） ：
【化１４】

【００１４】
［式 （２） において、Ｒ^１およびＲ^２の各々は水素原子であるか、あるいはそれらは一緒になって環外メチレンを形成し、Ｒ^３は単結合、メチレンまたはビニレンであり、Ｒ^４は直鎖状または分枝鎖状のＣ_１−Ｃ_７アルキル、アルケニル、アルコキシまたはアルキルアミノであり、Ｒ^５は水素原子またはメチルである。］
により表される化合物があげられる。
【００１５】
この中で、表２に示す化合物は好ましい。表２に示す化合物において、構造中に不斉炭素原子が存在する場合、化合物は（Ｓ） 配置および（Ｒ） 配置の両方を含む。Ｒ_３がビニレンであるとき、二重結合の立体配置は（Ｅ）配置および（Ｚ）配置の両方を含む。
【００１６】
【表２】

【００１７】
本発明の１つの目的は、血清中カルシウム濃度を増加させないでビタミンＤアンタゴニストで骨吸収を抑制することである。第２の目的は、骨パジェット病患者を治療するために本発明の組成物を使用することであり、ここで骨パジェット病患者の骨吸収活性は１α，２５−ジヒドロキシビタミンＤ_３の作用により極端に加速されている。
本発明者らは、本発明のビタミンＤアンタゴニストが、骨パジェット病患者の骨髄細胞からの１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞の形成を強く抑制し、そしてビタミンＤ欠乏ラットにおいて１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞の形成を抑制することを発見した。
【００１８】
上記化合物を有効成分として含有する骨パジェット病治療骨吸収抑制剤は、経口剤（軟カプセル剤、硬カプセル剤、錠剤またはシロップ剤）、または適当な賦形剤を使用して注射剤として製剤化することができる。例えば、骨粗しょう症患者を治療するために、経口剤は適切であるが、骨吸収が顕著に亢進した骨パジェット病患者において、注射剤が好ましい。注射剤はより大きいバイオアベイラビリティを有するにちがいない。
【００１９】
本発明において使用する非経口剤用の賦形剤は、例えば植物油、鉱物油、白色ワセリン、分岐鎖状の脂肪または油脂、高分子量アルコール等であろう。それらのなかで、例えば綿実油、トウモロコシ油、ココナッツ油またはアーモンド油のような植物油は好ましい。トリグリセリドの一部分として中鎖脂肪酸を含有する油は好ましい。
経口剤の賦形剤の好ましい例は、セルロース誘導体、例えば、結晶セルロース、ヒドロキシプロピルセルロース、ヒドロキシプロピルメチルセルロースまたはメチルセルロース、ポリビニルピロリドン、デキストリン、シクロデキストリン、カゼイン、ラクトース、マンニトール、ゼラチン等である。
【００２０】
本発明において骨吸収を抑制する有効成分の量はそれぞれ疾患の状態に応じて決められるが、一般的には、有効成分の量は０．００００４から０．２重量％、好ましくは０．０００１から０．１重量％である。
有効成分の投与量も、それぞれ患者の症状に応じて決められるが、一般的には、それは０．１から１０００ μｇ／日、好ましくは約１から１００ μｇ／日である。投与回数は通常１から３回／日である。これらの条件を満足するような方法で、製剤を調製することが好ましい。
【００２１】
実施例
以後実施例により本発明をさらに詳細に説明するが、本発明は実施例により限定されない。さらに、各実施例における化合物Ｎｏ．は上記表１に示す化合物Ｎｏ．である。
【００２２】
実施例 １．
正常のヒトの骨髄細胞からの １ α ，２５ −ジヒドロキシビタミン Ｄ _３ により惹起される破骨細胞形成に対する （２３Ｓ） − ２５ −デヒドロ− １ α ，２５ −ジヒドロキシビタミン Ｄ _３ − ２６，２３ −ラクトン ［ 化合物 １１ 、 （２３Ｓ 異性体 ）］ の破骨細胞形成抑制活性
Ｋｕｒｉｈａｒａ他の方法（Ｅｎｄｏｃｒｉｎｏｌｏｇｙ ｖｏｌ． １２６、２７３３−２７４１（１９９０）） に従い、健康な正常人の骨髄細胞から、単核細胞を分画した。簡単に述べると、骨髄細胞を正常人から獲得し、単核細胞画分をハイパーク−フィコール （Ｈｙｐａｑｕｅ−Ｆｉｃｏｌｌ） 密度勾配遠心により収集し、細胞画分をα−ミニマルエッセンシャルメディア（α−ＭＥＭ） で３回洗浄し、１０％の胎仔ウシ血清を含有するα−ＭＥＭの中に分散させた。この単核細胞懸濁液を１００ ｍｍの組織培養プレートの中に播種し、培養プレートを４％ＣＯ_２−空気雰囲気中で３７ ℃において９０分間保持し、非接着性細胞を収集した。２０％ウマ血清を含有するα−ＭＥＭの中に非接着性単核細胞を１０^６細胞／ｍｌで分散させ、細胞懸濁液を９６ウェルのマルチプレートの中に１００ μｌ／ウェルで播種した。
【００２３】
１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞形成を抑制する化合物１１（２３Ｓ異性体） の能力を次のようにして評価した：種々の濃度の１α，２５−ジヒドロキシビタミンＤ_３、種々の濃度の化合物１１（２３Ｓ異性体） または１０^{− ８}Ｍの１α，２５−ジヒドロキシビタミンＤ_３溶液と種々の濃度の化合物１１（２３Ｓ異性体） 溶液との組み合わせを各ウェルに添加した。培地を毎週取替え、４％ＣＯ_２−空気雰囲気中で３７ ℃において培養を３週間続けた。形成した破骨細胞を２３Ｃ６抗体に結合する能力により同定した。核をメチルグリーンで対比染色した。２３Ｃ６抗体と反応し、３またはそれ以上の核を有する細胞を骨芽細胞としてスコアを付けた。結果を表３に示す。
【００２４】
【表３】

【００２５】
１α，２５−ジヒドロキシビタミンＤ_３は、１０^−９から１０^−７ Ｍの濃度の１α，２５−ジヒドロキシビタミンＤ_３において濃度依存的に破骨細胞形成を惹起させた。破骨細胞形成は１０^−９Ｍの１α，２５−ジヒドロキシビタミンＤ_３において最大であった。化合物１１（２３Ｓ異性体） は破骨細胞形成を惹起しなかった。１０^−８ Ｍの１α，２５−ジヒドロキシビタミンＤ_３および種々の濃度の化合物１１（２３Ｓ異性体） を正常の骨髄培養物に同時に添加したとき、１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞形成は化合物１１（２３Ｓ異性体） により１０^−９から１０^−６ Ｍの濃度において抑制された。
【００２６】
実施例 ２．
骨パジェット病患者からの骨髄培養物において １ α ，２５ −ジヒドロキシビタミン Ｄ _３ により惹起される破骨細胞形成に対する （２３Ｓ） − ２５ −デヒドロ− １ α ，２５ −ジヒドロキシビタミン Ｄ _３ − ２６，２３ −ラクトン ［ 化合物 １１ 、 （２３Ｓ 異性体 ）］ の作用
骨髄細胞を骨パジェット病患者の関係する骨から獲得し、正常の骨細胞について前述したように処理し、培養した。培養物を実施例１に記載されているのと同一の方法で１α，２５−ジヒドロキシビタミンＤ_３および／または化合物１１（２３Ｓ異性体） で処理した。結果を表４に示す。
【００２７】
【表４】

【００２８】
骨パジェット病患者からの骨髄培養物において、１α，２５−ジヒドロキシビタミンＤ_３は１０^−１１ Ｍ程度に低い濃度において破骨細胞形成を惹起した。１α，２５−ジヒドロキシビタミンＤ_３は１０^−１１から１０^−７Ｍにおいて濃度依存的にこれらの培養物において破骨細胞形成を惹起した。破骨細胞形成はこれらの培養物において１０^−１０ Ｍと１０^−９ Ｍとの間で最大であった。これらは、正常の骨髄培養物における最大の破骨細胞形成に必要な濃度よりも１から２ ｌｏｇだけ低い、１α，２５−ジヒドロキシビタミンＤ_３濃度である。化合物１１（２３Ｓ異性体） はそれ自体破骨細胞形成を惹起せず、１α，２５−ジヒドロキシビタミンＤ_３の不存在下に、または１α，２５−ジヒドロキシビタミンＤ_３を添加したとき、破骨細胞形成を抑制した。
【００２９】
さらに、１０^−１０ Ｍの１α，２５−ジヒドロキシビタミンＤ_３を様々の濃度の化合物１１（２３Ｓ異性体） と組み合わせてこれらの培養物に添加したとき、破骨細胞形成は１０^−１１ Ｍと１０^−６ Ｍとの間の化合物１１（２３Ｓ異性体） 濃度において濃度依存的に抑制された。化合物１１（２３Ｓ異性体） は骨パジェット病患者の培養物における基底破骨細胞形成の増加を抑制し、これは正常の骨髄からの培養物において見られなかったことに注意すべきである。これらのデータが示唆するように、１α，２５−ジヒドロキシビタミンＤ_３の生理的濃度において、化合物１１（２３Ｓ異性体） は骨パジェット病患者において破骨細胞形成を正常レベルに戻すことができる。
【００３０】
実施例 ３．
骨パジェット病患者からの骨髄培養物における １ α ，２５ −ジヒドロキシビタミン Ｄ _３ により惹起される破骨細胞形成に対する様々のビタミン Ｄ アンタゴニストの破骨細胞抑制能力
実施例１に記載されているように骨髄培養を実施したが、ただし１α，２５−ジヒドロキシビタミンＤ_３で刺激した骨髄培養物に、様々のビタミンＤアンタゴニストを様々の濃度で添加した。骨髄細胞を骨パジェット病患者の関係する細胞から採取し、実施例２に記載されているように培養を実施した。化合物１３および１６において、２３位における不斉炭素原子に基づいて、２つのジアスレオマーが存在する。逆相ＨＰＬＣ上でより短い保持時間を有する異性体はより高い極性の異性体であり、そしてより長い保持時間を有する異性体はより低い極性の異性体であった。結果を表５に示す。
【００３１】
【表５】

【００３２】
表５に示すように、１α，２５−ジヒドロキシビタミンＤ_３は１０^−１１ Ｍから１０^−８ Ｍにおいて濃度依存的に破骨細胞形成を惹起した。化合物１１（２３Ｒ異性体）、化合物１３（より高い極性の異性体およびより低い極性の異性体） および化合物１６（より高い極性の異性体） 単独は破骨細胞形成を惹起せず、むしろ破骨細胞形成を抑制した。１０^−１０ Ｍの１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞形成は、化合物１１（２３Ｒ異性体）、化合物１３（より高い極性の異性体およびより低い極性の異性体） または化合物１６（より高い極性の異性体） により１０^−９ Ｍから１０^−７ Ｍにおいて濃度依存的に抑制された。１０^−７ Ｍの濃度において、１０^−１０ Ｍの１α，２５−ジヒドロキシビタミンＤ_３により惹起される破骨細胞形成はすべての化合物によりほとんど完全に抑制された。
【００３３】
実施例 ４．
骨パジェット病患者および同様な年齢の正常人の血液中のビタミン Ｄ 代謝物質の測定
血液試料（７から１０ ｍｌ）を９人の骨パジェット病患者および１０人の同様な年齢の正常人から収集し、血液試料を室温に３時間保持した。その後、それらを３０００ ｒｐｍで３０分間遠心して血清を得た。血清中のビタミンＤ代謝物質の濃度をＩｓｈｉｚｕｋａ他の方法により測定した（Ｊ． ｏｆ Ｎｕｔｒｉｔｉｏｎａｌ ａｎｄ Ｖｉｔａｍｉｎｏｌｏｇｙ、ｖｏｌ． ２７、７１−７５（１９８１） およびＡｃｔａ Ｅｎｄｏｃｒｉｎｏｌｏｇｙ、ｖｏｌ． １０４、９６−１０２（１９８３））。
【００３４】
簡単に述べると、３から５ ｍｌの各血清試料を水で１：３に希釈し、希釈した血清体積の２倍の体積のクロロホルム：メタノール（１：１） 混合溶液を添加し、この懸濁液を激しく震蘯させ、次いでクロロホルム層を収集した。水層をクロロホルムで再び抽出した。得られたクロロホルム層をプールし、ロータリーエバポレーターで濃縮し、残留物をセファデックス（Ｓｅｐｈａｄｅｘ） ＬＨ−２０カラム（１．２×１０ ｃｍ） にのせ、ｎ−ヘキサン：クロロホルム：メタノール（９：１：１） の混合溶媒で溶離した。８から１７ ｍｌの溶離液および１９から６０ ｍｌの溶離液を、それぞれ、２５−ＯＨ−Ｄ画分および２４，２５−（ＯＨ）_２Ｄ ＋ １α，２５−（ＯＨ）_２Ｄ画分として収集した。
【００３５】
さらに、２５−ＯＨ−Ｄ画分および２４，２５−（ＯＨ）_２Ｄ ＋ １α，２５−（ＯＨ）_２Ｄ画分の各々をゾルバックス（Ｚｏｒｂａｘ） ＳＩＬカラム（４．６×２５０ ｍｍ） にのせ、ｎ−ヘキサン中の１２％のイソプロパノールで溶離して２５−ＯＨ−Ｄ、２４，２５−（ＯＨ）_２Ｄおよび１α，２５−（ＯＨ）_２Ｄ画分を精製した。ビタミンＤ欠乏ラットからの血清の中に存在するビタミンＤ結合性タンパク質を使用する、競合タンパク質結合アッセイにより、精製された２５−ＯＨ−Ｄ画分および２４，２５−（ＯＨ）_２Ｄ画分の濃度を測定した。ビタミンＤ欠乏ヒヨコの小腸からのビタミンＤレセプターを使用する、放射性レセプターアッセイにより、精製された１α，２５−（ＯＨ）_２Ｄの濃度を測定した。結果を表６に示す。
【００３６】
【表６】

【００３７】
骨パジェット病患者からの血清中のビタミンＤ代謝物質濃度は、同様な年齢の正常人のものと有意に異ならなかった。ビタミンＤ代謝物質の異常性は骨パジェット病患者において検出されなかった。これらのデータが示すように、骨パジェット病患者からの血清中の１α，２５−（ＯＨ）_２Ｄの濃度は４１．０±９．１ ｐｇ／ｍｌ血清（１０^−１０ Ｍ）であった。
【００３８】
実施例１および２から明らかなように、骨パジェット病患者からの骨髄細胞による破骨細胞形成は１０^−１０ Ｍの１α，２５−ジヒドロキシビタミンＤ_３により惹起される。対照的に、正常人の骨髄細胞からの破骨細胞形成は１０^−１０ Ｍの１α，２５−ジヒドロキシビタミンＤ_３により惹起されなかった。Ｓｕｄａ他は、破骨細胞形成と骨吸収との間の陽性の相互関係が存在することを示した（Ｓｕｄａ、ＴおよびＴａｋａｈａｓｈｉ、Ｎ、Ｅｎｄｏｃｒｉｎｅ Ｒｅｖｉｅｗ 、ｖｏｌ． ２０、３４５−３５７（１９９９））。すなわち、破骨細胞形成を刺激する化合物はまた骨吸収活性に影響を与える。
【００３９】
実施例 ５．
ビタミン Ｄ 欠乏ラットにおける １ α ，２５ −ジヒドロキシビタミン Ｄ _３ により惹起される骨吸収に対する （２３Ｓ） − ２５ −デヒドロ− １ α ，２５ −ジヒドロキシビタミン Ｄ _３ − ２６，２３ −ラクトン ［ 化合物 １１ 、 （２３Ｓ 異性体 ）］ の骨吸収抑制活性
４週齢のウィスター系雄性ラットは、日本エスエルシー（ＳＬＣ、静岡） から購入したものを用いた。動物は、３匹ずつ金網ケージに入れ、動物飼育用ビタミンＤ欠乏飼料（Ｃａ、０．００３６％；Ｐ、０．３％；Ｈａｒｌａｎ Ｔａｋｌａｄ Ｒｅｓｅａｒｃｈ Ｄｉｅｔ、ＷＩ、Ｕ．Ｓ．Ａ．） および飲料水（０．４±０．２ ｐｐｍの次亜塩素酸塩で処理した井水） を７週間自由摂取させた。温度を２３±１ ℃、湿度を５５±１０％に保持した。用いた動物は、１実験群あたり５匹とした。
【００４０】
対照動物には溶媒（５％エタノール／Ｔｒｉｔｏｎ Ｘ−１００／生理的食塩水） を静脈内投与し、実験動物に１α，２５−ジヒドロキシビタミンＤ_３を０．２５ μｇ／ｋｇの投与量で静脈内投与した。ビタミンＤ誘導体を投与した動物について、２ μｇ／ｋｇ、１０ μｇ／ｋｇまたは５０ μｇ／ｋｇの投与量の化合物１１（２３Ｓ異性体） を投与するか、あるいは化合物１１（２３Ｓ異性体） 単独または化合物１１（２３Ｓ異性体） と０．２５ μｇ／ｋｇの投与量の１α，２５−ジヒドロキシビタミンＤ_３との組み合わせを投与した。体積は各動物について２ ｍｌ／ｋｇであった。投与２４時間後にエーテル麻酔下に腹部下降大動脈より採血し、定法にしたがって血清を得た後、血清中のカルシウム濃度を測定した。カルシウム濃度は日立製作所製７０７０型自動分析装置を用い、ＯＣＰＣ法（Ａｍ． Ｊ． Ｃｌｉｎ． Ｐａｔｈｏｌ． Ｖｏｌ． ４５、２９０−２９６（１９６６）） で測定した。結果を表７に示す。
【００４１】
【表７】

【００４２】
これらの結果が示すように、０．２５ μｇ／ｋｇの１α，２５−ジヒドロキシビタミンＤ_３の投与２４時間後に、動物は対照に比較して有意な血清中カルシウム濃度上昇を示した。これらのラットはカルシウムを含まない飼料で飼育したので、血清中カルシウム濃度上昇は破骨細胞による骨吸収により骨から放出されたカルシウムによる（Ａｍ． Ｊ． Ｃｌｉｎ． Ｐａｔｈｏｌ． Ｖｏｌ． ２１６、１３５１−１３５９（１９６９））。化合物１１（２３Ｓ異性体） を投与した動物について、血清中カルシウム濃度上昇は５０ μｇ／ｋｇの投与量においてさえ観測されなかった。したがって、この化合物は投与２４時間後に骨吸収を惹起しない。しかしながら、１α，２５−ジヒドロキシビタミンＤ_３と化合物１１（２３Ｓ異性体） を２ μｇ／ｋｇ、１０ μｇ／ｋｇまたは５０ μｇ／ｋｇの投与量で同時に投与したとき、骨吸収は上昇しなかった。これらのデータが証明するように、１α，２５−ジヒドロキシビタミンＤ_３により惹起される骨吸収は用量依存的に化合物１１（２３Ｓ異性体） の投与により抑制された。
【００４３】
以上の結果から、本発明のビタミンＤアンタゴニストはｉｎ ｖｉｖｏにおいて血清中カルシウム濃度を上昇させないで１α，２５−ジヒドロキシビタミンＤ_３により惹起される骨吸収を抑制することが明らかになった。従って、本発明のビタミンＤアンタゴニストは骨パジェット病患者において見られる１α，２５−ジヒドロキシビタミンＤ_３で惹起される骨吸収の亢進の治療に有用である。[0001]
[Field of the Invention]
The present invention relates to a group of therapeutic agents containing a vitamin D antagonist as an active ingredient for suppressing bone resorption. In particular, the invention relates to the use of these substances as therapeutic physiotherapeutics for Paget's disease of the bone.
[0002]
[Conventional technology]
Bone is a dynamic tissue that cycles through resorption and subsequent formation of new bone. This process continues through the skeleton in individual multicellular units called bone remodeling units. In adults, bone resorption and subsequent formation of new bone is called bone remodeling, and through the remodeling process, bone mass is maintained or increases or decreases over life. Relative bone mass depends on the balance between bone resorption level and bone formation level. In metabolic bone disease, the balance between this bone resorption and bone formation changes. Examples are osteoporosis and Paget's disease.
[0003]
Osteoporosis is a disease in which both the organic and mineral components of bone are reduced. In osteoporosis, bone mass is reduced and fractures are more likely. Osteoporosis is classified into two types, type I seen after menopause and type II seen in the elderly. In postmenopausal osteoporosis, also called type I osteoporosis, estrogen deficiency increases local levels of cytokines such as interleukin-6, interleukin-11 and interleukin-1. Treatment of postmenopausal osteoporosis includes, for example, administering estrogen or a derivative thereof, bisphosphonate, to block bone resorption by osteoclasts. Similarly, bone turnover activators, for example, 1.alpha.-hydroxy vitamin D, and l [alpha], 25-dihydroxyvitamin D ₃ prepared derivatives and osteogenic enhancers, for example, all of the vitamin K ₂ formulations have been used to treat this disease ing.
[0004]
Bone Paget's disease is the most severe example of abnormal bone remodeling. In Paget's disease of bone, bone resorption increases significantly, followed by the formation of abundant new bone. The bone formed is perturbed in structure, resulting in weakened bones that bend and break. The major cellular abnormality in bone Paget's disease lies in osteoclasts, the cells that resorb bone. In bone Paget's disease, osteoclasts contain paramyxovirus-like nuclear inclusions, suggesting that bone Paget's disease can be a slow disease caused by paramyxovirus. In addition, there may be a genetic component to Paget's disease of the bone. Up to 40% of these patients have a first degree relatively affected by the disease.
[0005]
Bone Paget's disease is the second most common bone disease following osteoporosis, and affects up to 2-3 million people in the United States. Bone Paget's disease is usually treated with bisphosphonate or calcitonin preparations that inhibit osteoclast activity and bone resorption. However, l [alpha] of osteoclast precursors in patients with Paget's disease, the presence of abnormality in the 25-dihydroxyvitamin _{D 3} sensitivity was suggested from recent discovery (J. Bone and Res. Vol. 15,228- 236 (2000)). We have found that osteoclast precursors from patients with Paget's disease of bone have osteoclasts at 1α, 25-dihydroxyvitamin D ₃ concentrations 1-2 logs lower than those required for normal osteoclastogenesis. Is formed. Furthermore, these studies showed that the increased sensitivity of 1α, 25-dihydroxyvitamin D ₃ appears to be mediated through the induction of activators of the vitamin D receptor.
[0006]
Compounds that inhibit the binding of 1α, 25-dihydroxyvitamin D _{3 to} its receptor or inhibit the action of 1α, 25-dihydroxyvitamin D _{3 on} osteoclast precursors are novel and have potential for treating Paget's disease of bone. Would be effective. If the blood levels of 1α, 25-dihydroxyvitamin D ₃ in patients with bone Paget's disease are normally similar, then these levels of 1α, 25-dihydroxyvitamin D ₃ are sufficient to cause bone resorption, which normally does not occur. Would. Thus, agents that block the effects of 1α, 25-dihydroxyvitamin D ₃ or hypersensitive osteoclast precursors in Paget's disease of bone would be very effective therapeutic physiotherapeutic agents. Such an agent would have advantages over current therapies for Paget's disease of the bone. For example, oral forms of bisphosphonates have significant gastrointestinal side effects, and the therapeutic effects of calcitonin ultimately diminish over time.
[0007]
Vitamin _{D 3} antagonists of the present invention, International Patent Publication WO 95/33716 Pat (compound of formula (1)), (compounds of formula (1)) No. WO 00/24712 specification, WO 94/07853 No. (Compounds of formula (2)) and WO 97/00242 (compounds of formula (2)). Compounds of formula (1) is, l [alpha], 25-dihydroxyvitamin _{D 3} and l [alpha], 25-dihydroxyvitamin _{D 3} and l [alpha], 25-bond between dihydroxyvitamin _{D 3} receptor (VDR) (J. Biol. Chem . 274, 16392-16399 (1999)), binding between VDR and 9-cis-retinoic acid receptor (RXR), and VDR and the transcriptional regulator steroid receptor coactivator 1 (SRC-1). coupling between (J. Biol. Chem., vol . 274,32376-32381 (1999)) by inhibiting, l [alpha], 25-suppressing direct action dihydroxyvitamin _{D 3.} Compounds of formula (2) is, l [alpha], antagonizes the action of 25-dihydroxyvitamin _{D 3.} See the following document: Biol. Chem. , Vol. 275, 16506-16512 (2000)).
[0008]
[Disclosure of the Invention]
The present inventors have discovered that vitamin D antagonists strongly inhibit the formation of 1α, 25-dihydroxyvitamin D ₃ -induced osteoclasts by normal bone marrow cells and bone marrow cells of patients with bone Paget's disease. . This reduction in osteoclast formation reduces bone resorption. The present inventors have also discovered that vitamin D antagonists suppress bone resorption induced by 1α, 25-dihydroxyvitamin D ₃ in vitamin D deficient animals, thereby making these antagonists in vitro and in vivo. It is proved that both have a high inhibitory effect on bone resorption.
Therefore, it is an object of the present invention to use a vitamin D antagonist to suppress bone resorption in patients with Paget's disease of the bone. These antagonists will suppress bone resorption without increasing serum calcium levels.
[0009]
[Best Mode for Carrying Out the Invention]
As a sample of the vitamin D antagonist of the present invention, the following formula (1):
Embedded image

[0010]
[In the formula (1), m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
A compound represented by the formula:
[0011]
Among them, compounds in which m is 1 or 2 are preferred. Further, with respect to the combination of m, q, r and X, the compounds shown in Table 1 are preferable. 11, 13, 16, 21, 23 and 26 are preferred. In the compounds shown in Table 1, when an asymmetric carbon atom is present in the structure, it includes both (S) and (R) configurations.
[0012]
[Table 1]

[0013]
Further, as a sample of the vitamin D antagonist of the present invention, the following formula (2):
Embedded image

[0014]
[In the formula (2), each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is straight or branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, R ⁵ is hydrogen atom or methyl. ]
A compound represented by the formula:
[0015]
Among them, the compounds shown in Table 2 are preferable. In the compounds shown in Table 2, when an asymmetric carbon atom is present in the structure, the compound contains both (S) configuration and (R) configuration. When R ₃ is vinylene, the configuration of the double bond includes both (E) and (Z) configurations.
[0016]
[Table 2]

[0017]
One object of the present invention is to suppress bone resorption with vitamin D antagonists without increasing serum calcium levels. A second objective is to use the compositions of the present invention to treat patients with bone Paget's disease, wherein the bone resorption activity of patients with bone Paget's disease is extreme due to the action of 1α, 25-dihydroxyvitamin D _3. Has been accelerated.
The present inventors have shown that the vitamin D antagonists of the present invention strongly inhibit the formation of 1α, 25-dihydroxyvitamin D ₃ -induced osteoclasts from bone marrow cells of patients with bone Paget's disease, and that vitamin D deficient rats Found that it inhibited the formation of osteoclasts induced by 1α, 25-dihydroxyvitamin D ₃ .
[0018]
The bone resorption inhibitor for treating Paget's disease of bone containing the above compound as an active ingredient is formulated into an oral preparation (soft capsule, hard capsule, tablet or syrup) or an injection using an appropriate excipient. can do. For example, an oral preparation is suitable for treating an osteoporosis patient, but an injection is preferable for a patient with bone Paget's disease in which bone resorption is significantly enhanced. Injectables must have greater bioavailability.
[0019]
Excipients for parenteral use in the present invention may be, for example, vegetable oils, mineral oils, white petrolatum, branched fats or oils, high molecular weight alcohols and the like. Among them, vegetable oils such as, for example, cottonseed oil, corn oil, coconut oil or almond oil are preferred. Oils containing medium chain fatty acids as part of the triglycerides are preferred.
Preferred examples of excipients for oral preparations are cellulose derivatives such as crystalline cellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose or methylcellulose, polyvinylpyrrolidone, dextrin, cyclodextrin, casein, lactose, mannitol, gelatin and the like.
[0020]
In the present invention, the amount of the active ingredient that inhibits bone resorption is determined depending on the disease state, but generally, the amount of the active ingredient is 0.00004 to 0.2% by weight, preferably 0.0001 to 0.2% by weight. 0.1% by weight.
The dosage of the active ingredient will also depend on the individual patient's condition, but will generally be from 0.1 to 1000 μg / day, preferably from about 1 to 100 μg / day. The frequency of administration is usually 1 to 3 times / day. It is preferable to prepare the preparation by a method that satisfies these conditions.
[0021]
Examples Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to the examples. Further, in each example, compound No. Is the compound No. shown in Table 1 above. It is.
[0022]
Embodiment 1 FIG .
1 alpha from normal human bone marrow cells, 25 - on osteoclast formation induced by dihydroxyvitamin D ₃ (23S) - 25 - dehydro - 1 alpha, 25 - dihydroxyvitamin D ₃ - 26, 23 - lactone [ Osteoclast formation inhibitory activity of compound 11 , (23S isomer )] Mononuclear cells are fractionated from bone marrow cells of healthy normal humans according to the method of Kurihara et al. (Endocrinology vol. 126, 2733-2741 (1990)). did. Briefly, bone marrow cells were obtained from a normal person, the mononuclear cell fraction was collected by Hyper-Ficoll density gradient centrifugation, and the cell fraction was collected using α-minimal essential media (α-MEM). , And dispersed in α-MEM containing 10% fetal calf serum. This mononuclear cell suspension was seeded in a 100 mm tissue culture plate, and the culture plate was kept in a 4% CO ₂ -air atmosphere at 37 ° C. for 90 minutes to collect non-adherent cells. Non-adherent mononuclear cells were dispersed at 10 ⁶ cells / ml in α-MEM containing 20% horse serum, and the cell suspension was seeded at 100 μl / well in a 96-well multiplate.
[0023]
The ability of compound 11 (23S isomer) to inhibit osteoclastogenesis induced by 1α, 25-dihydroxyvitamin D ₃ was evaluated as follows: various concentrations of 1α, 25-dihydroxyvitamin D ₃ , various concentrations of compound 11 (23S isomer) or 10 ^{- 8} M of l [alpha], a combination of 25-dihydroxyvitamin _{D 3} compound solutions with various concentrations 11 (23S isomer) was added to each well. The medium was changed weekly and the culture continued for 3 weeks at 37 ° C. in a 4% CO ₂ -air atmosphere. The formed osteoclasts were identified by their ability to bind to the 23C6 antibody. Nuclei were counterstained with methyl green. Cells that reacted with the 23C6 antibody and scored 3 or more nuclei were scored as osteoblasts. Table 3 shows the results.
[0024]
[Table 3]

[0025]
1α, 25-dihydroxyvitamin D ₃ induced osteoclastogenesis in a concentration-dependent manner at a concentration of 10 ⁻⁹ to 10 ⁻⁷ M of 1α, 25-dihydroxyvitamin D ₃ . Osteoclastogenesis was greatest in ^{10 -9} M of l [alpha], 25-dihydroxyvitamin _{D 3.} Compound 11 (23S isomer) did not induce osteoclast formation. 10 ^-8 M of l [alpha], 25-when added simultaneously dihydroxyvitamin _{D 3} and a variety of compounds 11 (23S isomer) The normal bone marrow cultures concentration, l [alpha], fracture induced by 25-dihydroxyvitamin _{D 3} Osteogenesis was inhibited by compound 11 (23S isomer) at a concentration of ^10-9 to ^10-6M .
[0026]
Embodiment 2 FIG .
1 alpha in bone marrow cultures from Paget's disease patients, 25 - on osteoclast formation induced by dihydroxyvitamin D ₃ (23S) - 25 - dehydro - 1 alpha, 25 - dihydroxyvitamin D ₃ - 26, 23 - Effect of lactone [ compound 11 , (23S isomer )] Bone marrow cells were obtained from the relevant bones of patients with bone Paget's disease, processed and cultured as described above for normal bone cells. Cultures were treated with 1α, 25-dihydroxyvitamin D ₃ and / or compound 11 (23S isomer) in the same manner as described in Example 1. Table 4 shows the results.
[0027]
[Table 4]

[0028]
In bone marrow cultures from patients with bone Paget's disease, 1α, 25-dihydroxyvitamin D ₃ induced osteoclast formation at concentrations as low as 10 ⁻¹¹ M. 1α, 25-dihydroxyvitamin D ₃ induced osteoclastogenesis in these cultures at 10 ⁻¹¹ to 10 ⁻⁷ M in a concentration-dependent manner. Osteoclast formation was maximal between 10 ⁻¹⁰ M and 10 ⁻⁹ M in these cultures. These are 1α, 25-dihydroxyvitamin D ₃ concentrations 1 to 2 logs lower than those required for maximal osteoclastogenesis in normal bone marrow cultures. Compound 11 (23S isomer) did not elicit itself osteoclastogenesis, l [alpha], 25-in the absence of a dihydroxyvitamin _{D 3,} or l [alpha], upon addition of 25-dihydroxy vitamin _{D 3,} osteoclast Formation was suppressed.
[0029]
In addition, when 10 ⁻¹⁰ M 1α, 25-dihydroxyvitamin D ₃ was added to these cultures in combination with various concentrations of compound 11 (23S isomer), osteoclastogenesis was 10 ⁻¹¹ M and 10 ⁻¹⁰ M. Compound 11 (23S isomer) concentration between ⁻⁶ M was suppressed in a concentration-dependent manner. It should be noted that compound 11 (23S isomer) suppressed increased basal osteoclastogenesis in cultures of Paget's disease patients, which was not seen in cultures from normal bone marrow. These data suggest that at physiological concentrations of 1α, 25-dihydroxyvitamin D ₃ , compound 11 (23S isomer) is able to return osteoclastogenesis to normal levels in patients with bone Paget's disease.
[0030]
Embodiment 3 FIG .
1 alpha, 25 in bone marrow cultures from Paget's disease patients - are in the osteoclast inhibiting ability <br/> Example 1 of the various vitamin D antagonists on osteoclastogenesis elicited by dihydroxyvitamin D ₃ It was performed bone marrow cultures as are, except l [alpha], bone marrow cultures stimulated with 25-dihydroxyvitamin D _3, was added various vitamin D antagonist at varying concentrations. Bone marrow cells were harvested from the relevant cells of a Paget's disease patient and cultured as described in Example 2. In compounds 13 and 16, there are two diastereomers based on the asymmetric carbon atom at position 23. The isomer with shorter retention time on reverse phase HPLC was the more polar isomer, and the isomer with longer retention time was the less polar isomer. Table 5 shows the results.
[0031]
[Table 5]

[0032]
As shown in Table 5, 1α, 25-dihydroxyvitamin D ₃ induced osteoclast formation in a concentration-dependent manner at 10 ⁻¹¹ M to 10 ⁻⁸ M. Compound 11 (23R isomer), Compound 13 (more polar and less polar isomers) and Compound 16 (more polar isomer) alone do not cause osteoclastogenesis, but rather osteoclasts Cell formation was suppressed. Osteoclastogenesis induced by 10 ⁻¹⁰ M 1α, 25-dihydroxyvitamin D ₃ is compound 11 (23R isomer), compound 13 (higher and lower polarity isomer) or compound 16 (the more polar isomer) suppressed in a concentration-dependent manner from 10 ⁻⁹ M to 10 ⁻⁷ M. At a concentration of 10 ⁻⁷ M, osteoclast formation induced by 10 ⁻¹⁰ M 1α, 25-dihydroxyvitamin D ₃ was almost completely suppressed by all compounds.
[0033]
Embodiment 4 FIG .
Determination of Vitamin D Metabolites in the Blood of Bone Paget's Disease Patients and Normals of Similar Age Blood samples (7 to 10 ml) were collected from 9 patients with Bone Paget's disease and 10 normal subjects of similar age. Blood samples were collected from humans and kept at room temperature for 3 hours. Thereafter, they were centrifuged at 3000 rpm for 30 minutes to obtain serum. The concentration of vitamin D metabolites in serum was determined by the method of Ishizuka et al. (J. of Nutrition and Vitamology, vol. 27, 71-75 (1981) and Acta Endocrinology, vol. 104, 96-102 (1983)). .
[0034]
Briefly, 3 to 5 ml of each serum sample was diluted 1: 3 with water, and a chloroform: methanol (1: 1) mixed solution of twice the diluted serum volume was added thereto. The solution was shaken vigorously and then the chloroform layer was collected. The aqueous layer was extracted again with chloroform. The resulting chloroform layers were pooled, concentrated on a rotary evaporator, and the residue was applied to a Sephadex LH-20 column (1.2 × 10 cm), and n-hexane: chloroform: methanol (9: 1: The mixture was eluted with the mixed solvent of 1). From 8 to 17 ml of the eluent and 19 60 ml eluent, respectively, 25-OH-D fraction and _{24,25- (OH) 2 D + 1α} , 25- (OH) collected as ₂ D fraction did.
[0035]
Furthermore, each of the 25-OH-D fraction and the 24,25- (OH) ₂ D + 1α, 25- (OH) ₂ D fraction is loaded on a Zorbax SIL column (4.6 × 250 mm). , 25-OH-D and eluted with 12% isopropanol in n- hexane, 24,25- _{(OH) 2} D and 1α, 25- _(OH) was purified ₂ D fraction. Use of vitamin D binding proteins present in serum from vitamin D-deficient rats, by a competitive protein binding assays, 25-OH-D fraction and purified _24,25-(OH) of ₂ D fraction The concentration was measured. The concentration of purified 1α, 25- (OH) ₂ D was determined by a radioreceptor assay using the vitamin D receptor from the small intestine of vitamin D-deficient chicks. Table 6 shows the results.
[0036]
[Table 6]

[0037]
Serum vitamin D metabolite levels from patients with bone Paget's disease did not differ significantly from that of normal individuals of similar age. No abnormalities of vitamin D metabolites were detected in patients with Paget's disease of bone. As these data indicate, the concentration of 1α, 25- (OH) ₂ D in serum from patients with bone Paget's disease was 41.0 ± 9.1 pg / ml serum (10 ⁻¹⁰ M).
[0038]
As apparent from Examples 1 and 2, osteoclast formation by bone marrow cells from bone Paget's disease patients of ^{10 -10} M l [alpha], elicited by 25-dihydroxyvitamin _{D 3.} In contrast, osteoclast formation from normal human bone marrow cells of ^{10 -10} M l [alpha], was not caused by 25-dihydroxyvitamin _{D 3.} Suda et al. Have shown that a positive correlation exists between osteoclastogenesis and bone resorption (Suda, T and Takahashi, N, Endocrine Review, vol. 20, 345-357 (1999)). That is, compounds that stimulate osteoclast formation also affect bone resorption activity.
[0039]
Embodiment 5 FIG .
1 alpha in vitamin D-deficient rats, 25 - (23S) on bone resorption induced by dihydroxyvitamin D ₃ - 25 - dehydro - 1 α, 25 - dihydroxyvitamin D ₃ - 26, 23 - lactone [compound 11, (23S Bone Resorption Inhibitory Activity of Isomers )] Wistar male rats of 4 weeks of age were purchased from Japan SLC (SLC, Shizuoka). The animals are housed in wire mesh cages three at a time, and are fed a vitamin D deficient diet for animal rearing (Ca, 0.0036%; P, 0.3%; Harlan Taklad Research Diet, WI, USA) and drinking water. (Well water treated with 0.4 ± 0.2 ppm of hypochlorite) was allowed to be taken freely for 7 weeks. The temperature was maintained at 23 ± 1 ° C. and the humidity at 55 ± 10%. Five animals were used per experimental group.
[0040]
The solvent in the control animals (5% ethanol / Triton X-100 / saline) was administered intravenously, l [alpha] in experimental animals, 25-dihydroxyvitamin _{D 3} intravenously at a dose of 0.25 [mu] g / kg Administration. For the animal to which the vitamin D derivative was administered, a dose of 2 μg / kg, 10 μg / kg or 50 μg / kg of compound 11 (23S isomer) was administered, or compound 11 (23S isomer) alone or compound 11 (23S isomer) and the dose of 0.25 [mu] g / kg of l [alpha], were administered a combination of a 25-dihydroxyvitamin _{D 3.} The volume was 2 ml / kg for each animal. Twenty-four hours after administration, blood was collected from the abdominal descending aorta under ether anesthesia, serum was obtained according to a standard method, and the calcium concentration in the serum was measured. The calcium concentration was measured by an OCPC method (Am. J. Clin. Pathol. Vol. 45, 290-296 (1966)) using an automatic analyzer 7070 manufactured by Hitachi, Ltd. Table 7 shows the results.
[0041]
[Table 7]

[0042]
As these results show, 24 hours after administration of 0.25 μg / kg of 1α, 25-dihydroxyvitamin D ₃ , animals showed a significant increase in serum calcium concentration compared to controls. Since these rats were kept on a calcium-free diet, the increase in serum calcium concentration was due to calcium released from bone by bone resorption by osteoclasts (Am. J. Clin. Pathol. Vol. 216, 1351-1359). (1969)). For animals receiving compound 11 (23S isomer), no increase in serum calcium concentration was observed even at a dose of 50 μg / kg. Thus, the compound does not cause bone resorption 24 hours after administration. However, when 1α, 25-dihydroxyvitamin D ₃ and compound 11 (23S isomer) were administered simultaneously at a dose of 2 μg / kg, 10 μg / kg or 50 μg / kg, bone resorption did not increase. These data demonstrate that bone resorption induced by 1α, 25-dihydroxyvitamin D ₃ was inhibited by administration of compound 11 (23S isomer) in a dose-dependent manner.
[0043]
From the above results, it was revealed that the vitamin D antagonist of the present invention suppressed bone resorption induced by 1α, 25-dihydroxyvitamin D ₃ without increasing serum calcium concentration in vivo. Therefore, vitamin D antagonist of the present invention is 1α seen in Paget's disease patients, are useful in the treatment of increased bone resorption induced by 25-dihydroxyvitamin D _3.

Claims (42)

A method of inhibiting bone resorption comprising administering a vitamin D antagonist to a patient. A method for treating Paget's disease of bone, comprising administering a vitamin D antagonist to a patient. The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
The method according to claim 1, which is a compound represented by the formula: The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
The method according to claim 2, which is a compound represented by the formula: 4. The method according to claim 3, wherein m is 1 or 2. 5. The method according to claim 4, wherein m is 1 or 2. 4. The method according to claim 3, wherein m is 1, q is 0, r is 1, and X is an oxygen atom. The method according to claim 4, wherein m is 1, q is 0, r is 1, and X is an oxygen atom. 4. The method of claim 3, wherein m is 1, q is 0, r is 1, and X is a carbon atom. 5. The method of claim 4, wherein m is 1, q is 0, r is 1, and X is a carbon atom. 4. The method according to claim 3, wherein m is 2, q is 0, r is 1, and X is an oxygen atom. 5. The method according to claim 4, wherein m is 2, q is 0, r is 1, and X is an oxygen atom. 4. The method of claim 3, wherein m is 1, q is 1, r is 0, and X is a carbon atom. 5. The method of claim 4, wherein m is 1, q is 1, r is 0, and X is a carbon atom. The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
The method according to claim 1, which is a compound represented by the formula: The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
The method according to claim 2, which is a compound represented by the formula: R ⁴ is, n- butyl, n- pentyl, n- hexyl, n- heptyl, 1-pentenyl, methoxy, ethoxy, n- propoxy, iso - propoxy, 2-methyl-propoxy, n- butoxy, t-butoxy, n The method according to claim 15, which is -pentyloxy, n-hexyloxy or n-heptyloxy, n-butylamino, n-pentylamino, n-heptylamino, n-pentenylamino. R ⁴ is, n- butyl, n- pentyl, n- hexyl, n- heptyl, 1-pentenyl, methoxy, ethoxy, n- propoxy, iso - propoxy, 2-methyl-propoxy, n- butoxy, t-butoxy, n 17. The method according to claim 16, which is -pentyloxy, n-hexyloxy or n-heptyloxy, n-butylamino, n-pentylamino, n-heptylamino, n-pentenylamino. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butoxy or 2-methylpropoxy, and R ⁵ is a hydrogen atom. The method described in. 17. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butoxy or 2-methylpropoxy, and R ⁵ is a hydrogen atom. The method described in. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butyl, n-pentyl, n-heptyl or 1-pentenyl, and R ⁵ is hydrogen 16. The method of claim 15, wherein the method is an atom. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butyl, n-pentyl, n-heptyl or 1-pentenyl, and R ⁵ is hydrogen 17. The method of claim 16, which is an atom. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butylamino, n-pentylamino, n-heptylamino or 1-pentenylamino; R ⁵ is a hydrogen atom, a method according to claim 15. R ¹ and R ² together form an exocyclic methylene, R ³ is a single bond, R ⁴ is n-butylamino, n-pentylamino, n-heptylamino or 1-pentenylamino; R ⁵ is a hydrogen atom, a method according to claim 16. The method according to claim 15, wherein R ¹ and R ² together form an exocyclic methylene, R ³ is vinylene, R ⁴ is ethoxy or t-butoxy, and R ⁵ is a hydrogen atom. . R ¹ and R ² together form a ring exocyclic methylene, R ³ is vinylene, R ⁴ is ethoxy or t- butoxy, R ⁵ is a hydrogen atom, A method according to claim 16 . The method according to claim 15, wherein each of R ¹ and R ² is hydrogen, R ³ is a single bond, R ⁴ is n-butoxy, and R ⁵ is a hydrogen atom. Each of R ¹ and ^{R 2} is hydrogen, ^{R 3} is a single bond, ^{R 4} is n- butoxy, ^{R 5} is a hydrogen atom, A method according to claim 16. The method according to claim 15, wherein each of R ¹ and R ² is hydrogen, R ³ is vinylene, R ⁴ is ethoxy, and R ⁵ is a hydrogen atom. Each of R ¹ and ^{R 2} are hydrogen, ^{R 3} is vinylene, ^{R 4} is ethoxy, ^{R 5} is a hydrogen atom, A method according to claim 16. A pharmaceutical composition for suppressing bone resorption, comprising a vitamin D antagonist. A pharmaceutical composition for treating Paget's disease of bone, comprising a vitamin D antagonist. The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
The pharmaceutical composition according to claim 31, which is a compound represented by the formula: The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
33. The pharmaceutical composition according to claim 32, which is a compound represented by the formula: The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
The pharmaceutical composition according to claim 31, which is a compound represented by the formula: The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
33. The pharmaceutical composition according to claim 32, which is a compound represented by the formula: Use of a vitamin D antagonist for the manufacture of a medicament for inhibiting bone resorption. Use of a vitamin D antagonist for the manufacture of a medicament for treating Paget's disease of the bone. The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
The use according to claim 37, which is a compound represented by the formula: The antagonist has the following formula (1):

[In the formula, m represents an integer of 1 to 3, q represents an integer of 0 to 3, r represents an integer of 0 to 3, and X represents a carbon atom or an oxygen atom. However, 1 ≦ q + r ≦ 3. ]
The use according to claim 38, which is a compound represented by the formula: The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
The use according to claim 37, which is a compound represented by the formula: The antagonist has the following formula (2):

Wherein each of R ¹ and R ² is a hydrogen atom or they together form an exocyclic methylene, R ³ is a single bond, methylene or vinylene, and R ⁴ is a linear Or a branched C ₁ -C ₇ alkyl, alkenyl, alkoxy or alkylamino, and R ⁵ is a hydrogen atom or methyl. ]
The use according to claim 38, which is a compound represented by the formula: