JPS58210017A - Antitumor agent - Google Patents

Abstract

PURPOSE:An orally administrable antitumor agent, containing 1alpha,25,26-trihydroxycholecalciferol as an active constituent, and capable of exhibiting cellulicidal effects on cancerous cells in vitro. CONSTITUTION:An antitumor agent containing 1alpha,25,26-trihydroxycholecalciferol expressed by the formula as an active constituent. The above-mentioned compound exhibits the inhibitory action on the multiplication of tumorous cells or cellulicidal effects on K-562 cells derived from human leukemias and LICR-LON- HMy2 cells derived from human myelomas in vitro and further antitumor effects found even in tests using mice and rats as hosts. The daily dose thereof is 0.1-10,000mug, preferably 0.5-1,000mug, for adults administered parenterally or orally. 1alpha,25S,26-Trihydroxycholecalciferol is preferred for the compound expressed by the formula.

Description

【発明の詳細な説明】 本発明は１α−，２５，２６−）リヒドロキシコレカル
シフエロールを含有する抗腫瘍剤に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an antitumor agent containing 1α-,25,26-)lyhydroxycholecalciferol.

現在用いられている抗悪性腫瘍剤としてはアルキル化剤
、代謝拮抗剤、抗生剤、４１ｈ物アルカロイド剤、免疫
療法剤等あるが、そのうち＠ｉ＃ｆｆｌ胞に対してｉｎ
　ｖｉｔｒｏにて殺細胞効果を示す薬剤は副作用も強い
ものが多い。Currently used antineoplastic agents include alkylating agents, antimetabolites, antibiotics, 41h alkaloids, and immunotherapeutic agents.
Many drugs that exhibit cell-killing effects in vitro also have strong side effects.

本発明者らは従来よシ生体内物質についての研究を行っ
てきた結果、１α、２５．２６−）リヒドロキシコレカ
ルシフエロール（以下、本％Ｉ　質又ハ１α、　２　Ｆ
＋　、　２６−（０Ｈ）３−Ｄｓと略称する）がｉｎ　
ｖｉｔｒ。The present inventors have previously conducted research on substances in the living body and found that 1α, 25.26-) lyhydroxycholecalciferol (hereinafter referred to as 1α, 26-)
+, 26-(0H)3-Ds) is in
vitr.

で癌細胞に対して殺細胞効果を示すことを知見した。It was found that this compound has a cytocidal effect on cancer cells.

本物質は次のような構造を有し、例えば「ビタミンＤ−
その新しい流れＪ（ＩＪ談社サすエンティフイク）　（
１９ｇ２）に開示されている。This substance has the following structure, for example, "vitamin D-
The new flow J (IJ Dansha Sasu Entifuku) (
19g2).

１（Ｘ、２５．２６−（ＯＨ）１−ＤＢ　　　ｌα、２
５８．２６　（０）［）Ｉ　ＤＡ本発明省らは、ｉｎ　
ｖｉＬｒｏでヒト自圧病由来のに−５６２、ヒト骨髄腫
由来のＬＩＣＲ−ＬＯＮ−ＩＩＭｙ２細胞を用いて本物
質の抗腫瘍効果を自べたところ臓＃Ｉ４細胞増殖抑制作
用或いは殺細胞効果が認められた。さらにマウス、ラッ
トを宿主として行った試験でも抗Ｍ瘍効果が耐められた
。1(X, 25.26-(OH)1-DB lα, 2
58.26 (0) [)I DA Ministry of the Invention, et al.
When viLro demonstrated the antitumor effect of this substance using Ni-562 cells derived from human autotonic disease and LICR-LON-IIMy2 cells derived from human myeloma, an inhibitory effect on the growth of viscera #I4 cells or a cell-killing effect was observed. Ta. Furthermore, the anti-M tumor effect was well tolerated in tests conducted using mice and rats as hosts.

本物質は１α＃　２５　Ｓ　−２６（ＯＨ）１−ＤＭ、
１α、２５Ｒ。This substance is 1α#25S-26(OH)1-DM,
1α, 25R.

２６−（ＯＨ）ｍ　Ｄａ又はその混合物であってもよい
が、特に１α、２５Ｓ、２６−（ＯＨ）畠−Ｄ、が好ま
しい。本発明の抗腫瘍剤は活性成分として上記の物質を
含有し、下記に示すごとき棹々の製剤形態で用いられる
。本発明の抗腫瘍剤は腹腔内尋の非経口的経路で投与さ
れるが経口的に投与され得る特徴を有する。26-(OH)mDa or a mixture thereof may be used, but 1α, 25S, and 26-(OH)Hatake-D are particularly preferred. The antitumor agent of the present invention contains the above-mentioned substances as active ingredients, and is used in various formulations as shown below. The antitumor agent of the present invention is administered parenterally via the intraperitoneal route, but has the characteristic that it can be administered orally.

本Ｖａ質を有効成分とする製剤は錠剤、散剤、顆粒剤、
坐剤、カプセル剤、アルコール溶液剤、油性溶液剤、水
性懸濁液剤などの投与形態で用いられる。又油性溶媒と
しては、中級脂肪酸のトリグリセライドエステル、コー
ン油、綿実油、落花生油、魚肝油、油状エステルなどが
用いられる。又カカオ油、グリセリン等も好ましい。そ
の他の成分として乳糖、でんぷん、メルク、ステアリン
酸マグネシウム、ソルビン酸、ソルビン酸の塩、糖又−
その誘導体、アルコール、生理食塩水、界面活性剤、酸
化防止剤等を本物質と併用１７得る。Preparations containing this Va quality as an active ingredient include tablets, powders, granules,
It is used in dosage forms such as suppositories, capsules, alcoholic solutions, oily solutions, and aqueous suspensions. As the oily solvent, triglyceride esters of intermediate fatty acids, corn oil, cottonseed oil, peanut oil, fish liver oil, oily esters, etc. are used. Also preferred are cacao oil and glycerin. Other ingredients include lactose, starch, Merck, magnesium stearate, sorbic acid, salts of sorbic acid, sugar and
Derivatives thereof, alcohol, physiological saline, surfactants, antioxidants, etc. are used in combination with this substance17.

本物質は、単位投与形態の中に０００００２〜４■μ％
、好ましくは０．０００２〜１雷１ｉｉ−％含有し得る
。又、本物質は成人に対し１日当り０．１μｑ〜１００
００μり、好−チしくｔま０，５〜１０００μノ投与す
る。This substance is present in unit dosage form at 000002-4μ%.
, preferably from 0.0002 to 1%. In addition, this substance is 0.1 μq to 100 μq per day for adults.
00 μm, preferably 0.5 to 1000 μm.

実施例１ ヒト白血病由来のに−５６２であり、１０％牛脂児血清
添加ＲＰＭＩ　１６４０培地に浮遊状で増殖するｉｎ　
ｖ１ｔｒｏ培養株を用いて実験を行った。それぞれの細
胞数がｌＸｌ０’／−となるように培地に懸濁させ、そ
の５−をシャーレに分注し、３７℃５％炭酸ガス含有空
気雰囲気の培養器中にて培養した。１α、　２５　Ｓ　
、　２６−　（ＯＩＩ）、−Ｄ、はジメチルスルホキシ
ド（以下ＤＭＳＯと略す）に溶解し、ＤＭｓ。Example 1 -562 is derived from human leukemia and grows in suspension in RPMI 1640 medium supplemented with 10% tallow serum.
Experiments were performed using the v1tro culture. Each cell was suspended in a medium so that the number of cells was 1X10'/-, and the 5- cells were dispensed into a Petri dish and cultured at 37°C in an incubator in an air atmosphere containing 5% carbon dioxide gas. 1α, 25 S
, 26-(OII), -D, was dissolved in dimethyl sulfoxide (hereinafter abbreviated as DMSO) to produce DMs.

の最終濃度が０．５容量％で１α、　２５８　、２６−
（ＯＨ）ｓ−Ｄ、が所定の濃度になるようにシャーレに
添加し、培誉３日後にトリバンプルー染色し、総生細胞
数を計測りまた。結果を第１表に示す。At a final concentration of 0.5% by volume, 1α, 258, 26-
(OH)s-D was added to a petri dish at a predetermined concentration, and after 3 days of culture, the cells were stained with trivan blue and the total number of viable cells was counted. The results are shown in Table 1.

第１表 増殖抑制率は浦媒（ＤＭＳＯ）投与群と比較した８３合
の夕ｇを示す。Table 1 shows the growth inhibition rate of 83 days compared to the DMSO administration group.

上記の如く、１α、　２５８　、２６−（ＯＨ）、−Ｄ
、は５゜ｎ９／ｍｌの濃度でに−５６２に対しては７７
％の細胞増殖抑制率を示した。As above, 1α, 258, 26-(OH), -D
, is 77 for −562 at a concentration of 5°n9/ml.
% cell growth inhibition rate.

実施例２ ヒトミニローマ由来ノＬＩＣＲ−ＬＯＮ−ＨＭｙ２であ
リ、１０％牛脂児血清添加ＲＰＭＩ　１６４０培地に浮
遊状で増殖するｉｎ　ｖｉｔｒｏ培養株を用いて実験を
行った。そわそれの細胞数がｌＸｌ０’／ｒｎｔとなる
ように培地に懸濁させ、その５　ｍｌをシャーレに分注
し、３７℃５％炭酸ガス含有空気雰囲気の培養器中にて
培養した。１α、　２５　Ｓ　、　２６−（Ｏｌｌ）、
−ＤｓけＤＭＳＯＫ溶解し、所定の濃度になるように添
加し、培養３日後にトリバンプルー染色ｊ７、総生細胞
数をｈ１測した。結果を第２表に示す。Example 2 An experiment was conducted using an in vitro culture strain of human miniroma-derived LICR-LON-HMy2 that grows in suspension in RPMI 1640 medium supplemented with 10% tallow serum. The fidget cells were suspended in a medium so that the number of cells was 1X10'/rnt, 5 ml of which was dispensed into a Petri dish, and cultured at 37°C in an incubator in an air atmosphere containing 5% carbon dioxide gas. 1α, 25S, 26-(Oll),
DMSOK was dissolved in Ds and added to a predetermined concentration, and after 3 days of culture, Trivan blue staining was carried out and the total number of living cells was counted. The results are shown in Table 2.

第２表 増殖抑制率はｈ媒（ＤＭＳ　Ｏ）投与群と比較した場合
の％を示す。Table 2 shows the growth inhibition rate as a percentage compared to the h vehicle (DMSO) administered group.

上記の如く、１α、　２５　Ｓ　、　２６−（ＯＨ）、
−Ｄ、は５０ｎり／ｒａｌの濃度でＬＩＣｆｔに対して
は８６％の細胞増殖抑制率を示した。As above, 1α, 25S, 26-(OH),
-D showed a cell proliferation inhibition rate of 86% against LICft at a concentration of 50n/ral.

実施例３ プルコン・バブリング中で４００Ｗ高圧水銀ランプで７
２時間照射して不純な反応性のパーオキシドを消失せし
めた中級脂肪酌のトリグリせライドニスデルＩ　Ｋｇに
ｌα、　２５　Ｓ　、　２６−（Ｏｎ（）ｓ−Ｄ、　５
　ｂＱを浴解し、１カプセル中にｌα、　２５８　、２
６−（Ｏｆ（）。Example 3 7 with 400W high pressure mercury lamp in pull-con bubbling
lα, 25 S, 26-(On()s-D, 5
bQ is dissolved in bath, lα, 258, 2 in one capsule.
6-(Of().

−Ｄ、を０．５μ（７含有するよりに下記剤皮成分を加
温俗解し秋カプセル製造機を用いで常法により軟カプセ
ル剤を作成した。Soft capsules containing 0.5 µm (7 μm) of -D were heated and soft capsules were prepared using an Aki capsule making machine in a conventional manner.

削成処方例 ピラチン　　　　　　１０垂鞘部 グリセリン　　　　２Ｎ 防腐剤（エチルパラベン）０．０５　　　ｔｔチタンホ
ワイト　０．２重量部 水　　　　　　０．２　　ｔｔ　　（最終形態に於ける
重量部）同様にしてｌカブ士ル中に１μｑ、２μｑ又は
５μすを含有するものをそれぞれ作成した。Example of abrasion recipe Piratin 10 parts Glycerin 2N Preservative (ethylparaben) 0.05 tt Titanium white 0.2 parts by weight Water 0.2 tt (parts by weight in final form) Similarly, in l Kabushiru. 1 μq, 2 μq, or 5 μl were prepared.

Claims (2)

【特許請求の範囲】[Claims] （１）１α、２５．２６−）リヒドロキシコレ力ルシフ
エロールを有効成分とする抗腫瘍剤。(1) An antitumor agent containing 1α, 25.26-)lyhydroxycholerylluciferol as an active ingredient. （２）１α、２５，２６−ドリヒドロキシコレカルシフ
エロールが１α、２５８．２６−１１ヒドロキシコレカ
ルシフエロールであることを特徴とする特許請求の範囲
第１項に記載の抗腫瘍剤。(2) The antitumor agent according to claim 1, wherein the 1α,25,26-dorihydroxycholecalciferol is 1α,258.26-11 hydroxycholecalciferol.