JPS58210020A - Antitumor agent - Google Patents

Abstract

PURPOSE:An orally administrable antitumor agent, containing 24S,25-dihydroxycholecalciferol as an active constituent, and capable of exhibiting cellulicidal effects on cancerous cells in vitro. CONSTITUTION:An antitumor agent containing 24S,25-dihydroxycholecalciferol 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.

Description

【発明の詳細な説明】 本発明は２４８．２５−：）ヒドロキシコレカルシフェ
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 antitumor agents containing 248.25-:)hydroxycholecalciferol.

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、植物アルカロイド９剤、免疫療
法剤等あるが、そのうち癌細胞に対してｉｎ　Ｖｉｔｒ
ｏ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, nine plant alkaloids, and immunotherapeutic agents.
Many of the drugs that exhibit cell-killing effects in o 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、２４８．２５−ジヒドロキシコレカルシフ
ェロールが１ｎｖ口ｒｏ　テｉ細胞に対して殺細胞効果
を示すことを知見した。The present inventors have previously conducted research on substances in vivo, and have found that 248.25-dihydroxycholecalciferol exhibits a cell-killing effect on 1nV rotary cells.

本物質は次のような構造を有し、例えばｋＮｏ、４０３
８２７２に開示されている。This substance has the following structure, for example kNo. 403
8272.

？Ｈ ２４８，２５−（ＯＨ）ｔ−Ｄａ 本発明者らは、ｉｎ　ｖｉｔｒｏでヒト白血病由来のＫ
　−５６２、？−）骨髄腫出来のＬＩＣＲ−ＬＯＮ−Ｍ
Ｈｙ２細胞を用いて本物質の抗腫瘍効果を詞べたところ
腫瘍細胞増殖抑制作用或いは殺細胞効果が認められた。? H248,25-(OH)t-Da The present inventors have demonstrated that human leukemia-derived K
-562,? -) LICR-LON-M caused by myeloma
When the antitumor effect of this substance was tested using Hy2 cells, tumor cell proliferation suppressive or cell killing effects were observed.

さらにマウス、ラットを宿主として行った試験でも抗腫
瘍効果が認められた。Furthermore, antitumor effects were observed in tests conducted using mice and rats as hosts.

本発明の抗腫瘍剤は活性成分として２４Ｓ、２５−ジヒ
ドロキシコレカルシフェロールを含有し、下記に示すご
とき種々の製剤形態で用いられる。本発明の抗腫瘍剤は
腹腔内等の非経口的紅路で投与されるが経口的に投与さ
れ得る特徴を有する。The antitumor agent of the present invention contains 24S, 25-dihydroxycholecalciferol as an active ingredient, and is used in various formulations as shown below. The antitumor agent of the present invention can be administered parenterally, such as intraperitoneally, but has the characteristic that it can be administered orally.

本物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９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、タルク、ステアリン
酸マグネシウム、ソルビン酸、ソルビン酸の塩、糖又は
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し得る。Preparations containing this substance as an active ingredient are used in dosage forms such as tablets, powders, granules, suppositories, capsules, alcoholic solutions, oily solutions, and aqueous suspensions. As the oily solvent, triglyceride 9 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 such as lactose, starch, talc, magnesium stearate, sorbic acid, salts of sorbic acid, sugar or derivatives thereof, alcohol, physiological saline, surfactants, antioxidants, etc. may be used in combination with this substance.

本物質は、単位投与形態の中に０．００００２〜４重ｉ
ｔチ、好甘しくけ０．０００２〜１重量％含有し得る。The substance may be present in unit dosage form from 0.00002 to 4 times i.
It may preferably contain 0.0002 to 1% by weight.

又、本物質は成人に対し１日当ｆｉ０．１μＶ〜１０，
０００μｔ１　好ましくは０．５〜１，０００μ？投与
する。In addition, this substance has a daily fi of 0.1 μV to 10,
000μt1 Preferably 0.5 to 1,000μ? Administer.

実施例１ ヒト白血病由来のＫ　−５６２であり、１０チ牛脂児血
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。それぞれの
細胞数がＩ　Ｘ　１０”／ｒｎｌとなるように培地に懸
濁させ、その５ｍｌをシャーレに分注し、３７℃５チ炭
酸ガス含有空気雰囲気の培養器中にて培養した。Example 1 K-562 derived from human leukemia, grown in suspension in RPMI 1640 medium supplemented with 10 times tallow serum.
Experiments were performed using vitro cultured strains. Each cell was suspended in a medium to a number of I x 10''/rnl, 5 ml of which was dispensed into a petri dish, and cultured at 37° C. in an incubator in an air atmosphere containing carbon dioxide gas.

２４８　、２５−　（ＯＨ）ｇ　−Ｄｓはジメチルスル
ホキシド９（以下ＤＭＳＯと略す）に溶解し、ＤＭＳ　
Ｏの最終濃度が０．５容量係で２４８　、２５−　（Ｏ
Ｈ）＊−Ｄ３が所定の濃度になるようにシャーレに添加
し、培養３日後にトリバンプルー染色し、総生細胞数を
計測した。結果を第１表に示す。248, 25-(OH)g-Ds was dissolved in dimethyl sulfoxide 9 (hereinafter abbreviated as DMSO), and DMS
When the final concentration of O is 0.5 volume, 248, 25- (O
H)*-D3 was added to a Petri dish at a predetermined concentration, and after 3 days of culture, it was stained with Trivan blue and the total number of living cells was counted. The results are shown in Table 1.

増殖抑制率は溶媒（ＤＭＳＯ）投与群と比較した場合の
俤を示す。The growth inhibition rate shows the increase when compared with the vehicle (DMSO) administration group.

上記の如く、２４８　、２５−　（ＯＨ）ｔ−Ｄｓは２
００バ／ｒｎｌの８度でＫ　−５６２に対しては６６憾
の細胞増殖抑制率を示した。As mentioned above, 248,25-(OH)t-Ds is 2
It showed a cell proliferation inhibition rate of 66% against K-562 at 8 degrees of 00 bar/rnl.

実施例２ ヒトミエローマ由来のＬＩＣＲ−ＬＯＮ−ＨＭｙ２であ
υ、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。それぞれの細胞数がＩ　Ｘ　ｌＯ”７ｍｌとな
る、ように培地に懸濁させ、その５ｔｎｌをシャーレに
分注し、３７℃５チ炭酸ガス含有空気雰囲気の培養器中
にて培養した。２４８　、２５−　（ＯＨ）ｔ　−Ｄｓ
はＤＭＳＯに浴解し、所定の濃度になるように添加し、
培養３日後にトリバンブルー染色し、総生細胞数を「［
測した。結果を第２衣に示す。Example 2 An experiment was conducted using an in vitro culture strain of LICR-LON-HMy2 derived from human myeloma, which grows in suspension in RPMI 1640 medium supplemented with 10% tallow serum. Each cell was suspended in a medium so that the number of cells was 7 ml, and 5 tnl of the suspension was dispensed into a petri dish and cultured at 37°C in an incubator with an air atmosphere containing carbon dioxide gas.248 25-(OH)t-Ds
is dissolved in DMSO and added to a predetermined concentration,
After 3 days of culture, the total number of viable cells was determined by staining with Trivan blue.
I measured it. The results are shown in the second layer.

第　　　　２　　　　表 増殖抑制率は溶媒（ＤＭＳＯ）投与群と比較した場合の
俤を示す。Table 2 shows the growth inhibition rate when compared with the vehicle (DMSO) administration group.

上記の如く、２４　Ｓ　、　２５　　（ＯＨ）ｔ　−’
Ｄａは２００ｐ？汐ｔの濃度でＬＩＣＲに対しては７３
％の細胞増殖抑制率を示した。As above, 24 S , 25 (OH)t −'
Da is 200p? 73 for LICR at the concentration of Shio-t.
% cell growth inhibition rate.

実施例３ アルゴン・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ンドを消失せし
めた中級脂肪酸のトリグリセライビエステル１ｋｇに２
４８，２５−（ＯＨ）ｔ−Ｄａ　ｓｉｗを溶解し、１カ
プセル中に２４８．２５−（ＯＨ）＊−Ｄｓを０．５μ
ｉ含有するように下記剤皮成分を加温溶解し軟カプセル
製造機を用いて常法によシ軟カプセル剤を作成した。Example 3 7 with 400W high pressure mercury lamp in argon bubbling
2 for 1 kg of triglycerai biester of intermediate fatty acid which has been irradiated for 2 hours to eliminate impure reactive perokind.
Dissolve 48,25-(OH)t-Da siw and add 0.5μ of 248.25-(OH)*-Ds in one capsule.
Soft capsules were prepared by heating and dissolving the following shell components so as to contain i and using a soft capsule making machine in a conventional manner.

剤皮処方例 ゼ　　ラ　　チ　　ン　　　　　１０　重量部グ　リ　
　セ　　リ　　ン　　　　　　　２　　　〃防腐剤（エ
チノＬ＝−９うＲン）　　　　０．０５／／チタンホワ
イト　　　０．２〃 水　　　　　　　　０．２〃（最終形態に於ける重量部
） 同様にして１カプセル中に１μ？、　　２μＶ又は５μ
？含有するものをそれぞれ作成した。Shell formulation example Gelatin 10 parts by weight
Serine 2 Preservative (Ethino L=-9R) 0.05//Titanium White 0.2 Water 0.2 (parts by weight in final form) Similarly, 1μ in 1 capsule ? , 2μV or 5μ
? We created each of the items that contain them.

１いｊす・　　　川　　　１−１　　　　義　　　左１
ｆ＝９５−1st river 1-1 righteousness left 1
f=95-

Claims (1)

【特許請求の範囲】[Claims] ＋１１　２４ｓ、２５−ｙヒドロキシコレカルシフェロ
ールを有効成分とする抗腫瘍剤。+11 An antitumor agent containing 24s, 25-y hydroxycholecalciferol as an active ingredient.