JPH0840908A - Cancerous cell proliferation inhibitor - Google Patents

Abstract

PURPOSE:To obtain a cancerous cell proliferation inhibltor or medicine for cancer presenting medicinal virtues to inhibit ligand-dependent cell proliferation such as for CCK-8 or gastrin, containing, as active ingredient, a specific benzodiazepine derivative or a pharmaceutically permissible salt thereof. CONSTITUTION:This cancerous cell proliferation inhibitor or medicine for cancer contains, as active ingredient, a benzodiazepine derivative of the formula (R<1> is an aryl, five-membered monocyclc, six-membered monocyclic or five- membered-six-membered condensed aromatic heterocyclic group; R<2> is an aryl) or a pharmaceutically permissible salt thereof. The compound of the formula is esp. (R)-1-[2,3-dihydro-1-(2'-methylphenacyl)-2-oxo-5-phenyl-1H-1,4- benzodiazepin-3-yl]-3-(3-tolyl) urea or a pharmaceutically permissible salt thereof. This active ingredient, which inhibit rapid proliferation to be termed a characteristic of cancerous cell, is useful as a medicine for pulmonary cancer, pancreatic cancer, colon cancer, gastric cancer, T-cell lymphoma, etc., being also useful as a cancer diagnostic.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a compound of general formula

Embedded image (The symbols in the formulas have the following meanings: R ¹ : an optionally substituted aryl group or a 5-membered monocycle,
A 6-membered monocyclic or 5-membered to 6-membered continuous aromatic heterocyclic group. R ^2: an aryl group which may be substituted. ), A benzodiazepine derivative or a pharmaceutically acceptable salt thereof as an active ingredient, and a cancer cell growth inhibitor and a cancer therapeutic agent.

[0002]

2. Description of the Related Art The compound represented by the general formula (I) or a salt thereof, which is an active ingredient of the medicament of the present invention, is CCK-B (cholecystokinin-Brai).
n) Receptor and gastrin receptor antagonistic action is known to be useful for digestive system diseases such as gastric ulcer, duodenal ulcer, gastritis and disorders of central nervous system such as appetite regulation system, pain and anxiety. (International Publication No. 92/11246
Issue pamphlet (1992)). Human cerebral CCK-B receptor and gastrin receptor have been shown to be identical by gene cloning (J. Biol. Chem. 268 , 18300 (199).
3), J. Biol. Chem. 268 , 8164 (1993)), lung cancer, pancreatic cancer, colon cancer, gastric cancer, T-cell lymphoma, and many other human cancer cell lines. These human cancer cell lines, are known to grow a concentration-dependent manner by a ligand such as CCK or gastrin (Am.J.Physiol. 266, 277 (1994 ), Cancer Re
s. 53 , 5208 (1993), Cancer Res. 49 , 2840 (1989), Oncogene
9 , 861 (1994)). On the other hand, L-365260 (a compound described in US Pat. No. 4,820,834, Example 281), which is a selective CCK-B / gastrin receptor antagonist, was used in a cell line derived from human small cell lung cancer. , CCK
Has been reported to inhibit the specific binding of CCK-8, which is a partial peptide showing biological activity of Peptides (Peptides
8 , 103 (1987), Peptides 11 , 1033 (1990)), the binding inhibitory activity of this compound on the CCK-B / gastrin receptor is weak, and it has not been clinically applied as a therapeutic drug for cancer.

[0003]

The present inventors have found that the CCK-
As a result of intensive studies on compounds represented by the above general formula (I) having B / gastrin receptor antagonistic action, they found that these compounds suppress ligand-dependent cell growth of CCK-8, gastrin and the like. The present invention has been completed. That is, the present invention relates to a CCK-B / gastrin receptor-expressing cancer cell growth inhibitor containing a compound represented by the general formula (I) or a salt thereof as an active ingredient. In particular,
An object of the present invention is to provide a drug containing a compound that binds to CCK-B / gastrin receptor expressed in cancer cells and antagonizes the actions of CCK and gastrin. Furthermore, the compound represented by the general formula (I), which is an active ingredient of the present invention, also suppresses DNA synthesis in cancer cells expressing CCK-B / gastrin receptor and exhibits a cancer cell growth inhibitory action.

Examples of the "aryl group" in the compound represented by formula (I) include a phenyl group, an indenyl group and a naphthyl group. These aryl groups may have a substituent. Examples of this substituent include a lower alkyl group, a lower alkoxy group and a halogen atom. Here, the lower alkyl group is a linear or branched hydrocarbon chain having 1 to 6 carbon atoms, and typical examples thereof include a methyl group, an ethyl group, a propyl group, and n-butyl. Group, n-pentyl group, isopropyl group, sec-butyl group and the like. The lower alkoxy group is an alkoxy group having the above lower alkyl group, and examples thereof include a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, and an n-butoxy group. Examples of the halogen atom include a fluorine atom, a chlorine atom or a bromine atom. Next, as the "5-membered monocyclic, 6-membered monocyclic or 5-membered-6-membered aromatic heterocyclic group", a thiophene ring, a furan ring, a pyrrole ring, a thiazole ring, an oxazole ring, an imidazole ring, a pyridine ring, Examples thereof include a benzothiophene ring, a benzofuran ring and an indole ring.

Next, since the compound (I), which is the active ingredient of the present invention, has an asymmetric carbon atom and has isomers, these isomers are also adopted as the active ingredient of the present invention. be able to. Further, the compound of the present invention forms a salt with an acid. Examples of such salts include salts with inorganic acids or organic acids such as hydrochlorides, sulfates and acetates. In addition, about the manufacturing method of compound (I) and its salt,
International Publication No. 92/11246 Pamphlet (199
It is described in the application pertaining to 2).

[0006]

INDUSTRIAL APPLICABILITY The compound represented by the general formula (I) or a salt thereof, which is an active ingredient of the medicament of the present invention, suppresses rapid proliferation, which can be said to be characteristic of cancer cells, and therefore lung cancer, pancreatic cancer, colon cancer, gastric cancer It is useful as a therapeutic drug for cancer such as T cell lymphoma. Furthermore, the CCK-B of the cancer cells / the gastrin receptor is inhibited by the ligand binding inhibitory action to the C of the cancer cells.
It is also possible to detect sensitivity to CK and gastrin, and it is also useful as a cancer diagnostic agent.

The compound of the present invention has low toxicity and is suitable for use as a medicine. For example, in the SD rat test, no serious side effect was observed even when 100 mg / kg was orally administered. The compound of the present invention or a salt thereof is adjusted to tablets, powders, fine granules, capsules, pills, solutions, injections, suppositories, ointments, patches and the like, and is orally (including sublingual administration) or parenteral. Will be given to you. Examples of the simple substance or excipient for the preparation include solid or liquid non-toxic medicinal substances. Examples of these are lactose, magnesium stearate, starch, talc, gelatin,
Agar, pectin, gum arabic, olive oil, sesame oil,
Examples include cocoa butter, ethylene glycol, and other commonly used ones. The clinical dose of the compound of the present invention is appropriately determined in consideration of the symptoms, weight, age, sex, etc. of the patient to whom it is applied, but it is usually 1 to 10 by oral administration per day for an adult.
The dose is 00 mg, which is to be administered once or in several divided doses.

<Experimental Examples and Examples> The present invention will be further described with reference to Experimental Examples and Examples. Experimental Example 1 (Human Cancer Cell Proliferation Inhibitory Action) Significance of Experimental System The cell proliferation inhibitory action was measured using DNA synthesis as an index by measuring the uptake of thymidine, which is a precursor of DNA, into cells. In the following Experimental Examples and Examples, (R) -1- [2,3-dihydro-1- (2′-methylphenacyl) -2-oxo among the compounds represented by the general formula (I) was used as a test compound. -5-phenyl-1H-1,
4-Benzodiazepin-3-yl] -3- (3-tolyl) urea (hereinafter referred to as Compound A) was used as a control compound thereof in L-365260 (US Pat. No. 4,820,
No. 834, the compound described in Example 281) was adopted.

Experimental Method NCIH510A cells derived from human small cell lung cancer were 10%
RPMI 1640 medium supplemented with fetal bovine serum (Gibuco
Cultured and maintained in Grand Island, NY) at 37 ° C. under 5% CO ₂ was used. 96 cells above serum-starved
Seed on a well plate, add gastrin (10 ^-8 M) and various concentrations of test compound or control compound to RPMI medium supplemented with insulin and transferrin, and incubate for 44 hours, followed by methyl [ ³ H] thymidine (1 Ciml , 25mCi / m
mol, Amersham) was added and the cells were cultured for 4 hours. After culturing, trichloroacetic acid (5%) was added to remove thymidine that was not incorporated by centrifugation, and the precipitate (cells) was lysed with 1 mM SDS-added 1 mM sodium hydroxide. The amount of [ ³ H] was measured with a liquid scintillation counter and used as the amount of thymidine incorporated into cells. The result is shown in FIG. From this result, it was revealed that the test compound significantly inhibited the growth of cancer cells because it inhibited thymidine incorporation in DNA synthesis at 10 ⁻⁷ M or more. On the other hand, the control compound L-365260 suppressed the thymidine uptake to the same extent with a 10-fold amount of the test compound.
It was found to be 10 times more effective than the control compound.

Experimental Example 2 (Inhibition of proliferation of human CCK-B / gastrin receptor-expressing mouse cells) Significance of the experimental system The cell proliferation promoting action of CCK and gastrin is CCK-B / in which CCK or gastrin is present on the cell surface. By binding to the gastrin receptor, the intracellular signal transduction mechanism via inositol phosphate, intracellular Ca ^2+, etc. is activated, and the amplified signal reaches the nucleus and DN in the nucleus is activated.
It is believed that the promotion of A synthesis leads to cell proliferation. Therefore, using mouse fibroblasts (non-cancerous cells) expressing human CCK-B / gastrin receptor, CCK of CCK by the compound of the present invention is used with the production of inositol phosphate, which is an intracellular signal transduction factor, as an index. -Inhibition of binding to B / gastrin receptor-expressing cells, and accompanying suppression of activation of intracellular signal transduction mechanism and suppression of cell proliferation were confirmed. Here, CCK-8 used in the following experiments is
It is a partial peptide of CCK and has 8 amino acid residues from 26 to 33, and exhibits a ligand action of CCK-B / gastrin receptor (Yalow: Gastroenterology. 58 , 609 (197
0)).

Experimental Method Mouse CHI-B / gastrin receptor-expressing mouse fibroblast cell line NIH3T3 cells (Taniguchi et al .: Oncog
ene 9, 861 (1994)) 10% bovine serum (ICN Flow, Costa Me
sa, CA) added DMEM medium (Gibuco, Grand Islan
d, NY) were used after being maintained in culture. The serum-starved cells were treated with myo [ ³ H] inositol (1 μCiml, 10
^-20 mCi / mmol, Amersham) was preliminarily labeled for 24 hours.
After washing the cells twice with DMEM medium, CCK-8 (5
X 10 ^-9 M) and various concentrations of the test compound or control compound were added, and the mixture was incubated at 37 ° C in the presence of 10 mM chlorolithium for 30 minutes.
A minute reaction was performed. The reaction was stopped by the addition of ice-cooled methanol, inositol phosphate was extracted, and the [ ³ H] amount was measured by a liquid scintillation counter to determine the amount of inositol phosphate produced. The result is shown in FIG. From these results, the compound represented by the general formula (I), which is the active ingredient of the pharmaceutical composition of the present invention, showed a significantly stronger inositol phosphate production inhibitory effect than the control compound. That is, the compound A showed 90% or more inhibition at the amount of 10 ^-8 M, whereas the control compound did not show the inhibition effect. As a result, the compound (I), which is the active ingredient of the pharmaceutical composition of the present invention, was identified as human CCK-B
/ Gastrin receptor-expressing cells showed remarkable inhibition of activation of intracellular signal transduction mechanism. Therefore, the cell growth inhibitory effect is inhibition of binding to CCK-B / gastrin receptor-expressing cells and accompanying intracellular signal transduction. It was confirmed that it was due to suppression of activation of the mechanism.

[0012]

EXAMPLES The medicine of the present invention will be described in more detail below with reference to examples. Example 1 (tablet) composition 20 mg tablet 40 mg tablet compound A 20 mg 40 mg lactose 73.4 80 corn starch 18 20 hydroxypropylcellulose 45 carboxymethylcellulose calcium 4 4.2 magnesium stearate 0.6 0.8 total 120 mg 150 mg

Preparation Method of 20 mg Tablet Compound A (100 g), lactose (367 g) and corn starch (90 g) were uniformly mixed using a fluidized granulation coating apparatus (manufactured by Okawara Seisakusho). 200 g of a 10% hydroxypropyl cellulose solution was sprayed on this and granulated. After drying, it is passed through a 20-mesh sieve, 20 g of carboxymethylcellulose calcium and 3 g of magnesium stearate are added, and 120 mg per tablet is obtained using a rotary tableting machine (manufactured by Hata Tekko Co., Ltd.) with a 7 mm x 8.4 R pestle. As tablets.

Preparation Method of 40 mg Tablet Compound A (140 g), lactose (280 g) and corn starch (70 g) were uniformly mixed using a fluidized granulation coating apparatus (manufactured by Okawara Seisakusho). 175 g of a 10% hydroxypropyl cellulose solution was sprayed on this and granulated. After drying, it was passed through a 20-mesh sieve, 14.7 g of carboxymethylcellulose calcium and 2.8 g of magnesium stearate were added thereto, and a rotary tableting machine (manufactured by Hata Tekko Co., Ltd.) was used to use a 7.5 mm × 9R pestle. 1
Each tablet was 150 mg.

[Brief description of drawings]

FIG. 1 is a diagram comparing the amount of thymidine incorporated into cells for Compound A and a control compound.

FIG. 2 is a diagram comparing the amount of inositol phosphate produced for compound A and a control compound.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Taizo Taniguchi 5-10-1-801 Hinomine, Kita-ku, Kobe-shi, Hyogo (72) Inventor Nobu Akira Iwata 14-8-208 Sumiyoshidai, Higashinada-ku, Kobe-shi, Hyogo ( 72) Inventor Toru Murayama 5-2-7-302 Tamon-dori, Chuo-ku, Kobe-shi, Hyogo Prefecture (72) Masato Sato 9 Rumi Tsukuba 321 9-5 2-chome Ninomiya, Tsukuba, Ibaraki

Claims (3)

[Claims] 1. A compound of the general formula (The symbols in the formulas have the following meanings: R ¹ : an optionally substituted aryl group or a 5-membered monocycle,
A 6-membered monocyclic or 5-membered to 6-membered continuous aromatic heterocyclic group. R ^2: an aryl group which may be substituted. ) A cancer cell growth inhibitor comprising a benzodiazepine derivative or a pharmaceutically acceptable salt thereof as an active ingredient. 2. (R) -1- [2,3-dihydro-1-
(2'-Methylphenacyl) -2-oxo-5-phenyl-1H-1,4-benzodiazepin-3-yl] -3
The cancer cell proliferation inhibitor according to claim 1, which comprises-(3-tolyl) urea or a pharmaceutically acceptable salt thereof as an active ingredient. 3. The cancer cell growth inhibitor according to claim 1, which is used as a therapeutic drug for cancer.