JPS5959617A - Anticancer medicine composition - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel pharmaceutical composition having anticancer activity.

Yoneba Patent No. 8.7418.856 1-
It has been disclosed that benzoyl-3-phenylureas have insecticidal activity. Its effects are influenced by mechanisms that are currently not fully understood. I learned that the reason why these compounds kill insects is because these [-benzoyl-3=phenylureas] influence chitin production, that is, suppress it.

In addition, European special γ “Exit number 1 +1,025,3
No. 63 describes that N-benzoyl-Nl-bilysiloxyphenylurea has an anticancer effect.

Among the 1-benzoyl-3-phenylureas known per se, European Patent Application No. 06025°863
No., Special Publication No. 109721 and Special Publication No. 1987-
8+664 excluding the compound having anticancer effect 1 (these have an inhibitory effect on chitin production) and these l! We have now obtained the finding that a metabolite has an extremely interesting anti-tumor effect on mammalian tumors by inhibiting the growth of Il+ cells.

To put it in more detail, the following 1) L-(2,6-difluorobenzoyl)-3-(
4-chlorophenyl) urea (also known as diflubenzuron)! 2)1. -(2+6-difluberonzol)-3-[
a, 5-dichloro-4-(8-chloro-5-1-lifluoromethyl-2-bilinroxy)phenyl]urea+
13) 1-(2,6
-difluorobenzoyl)-3-[3-chloro-4-(
4-chlorophenoxy)phenyl]urea; 4) L-(2,6-difluorobenzoyl)-3-(4
-) IJ fluoromedylphenyl) Urine 2'd (PH
5) L-(2-chlorobenzoyl)-3-14-chlorophenyl)urea 6) L-(2-chlorobenzoyl)-3-(41-trifluorometocyphenyl)urea寥7) L −(2
,c,-difluorobenzoyl)-3-(4-rioo-
2-Hydroxyphe= /l/ >by; Xl 8) l-(2,6-difluorobenzoyl)-3-(4
-Chloro-3-hydroxyphenyl)urea "9) L-(2-chlorobenzoyl)-3-(2-hydroxy-4-trifluoromethoxyphenyl)urea; 0) L-(2,6-difluorobenzoyl)- 3-(4
-cyanophenyl) urinary body 2 LL) L-(2,6-difluorobenzoyl)-8-C
4-(L, 172.3,8.3-hexafluoropropoxy)phenyl)urine a+ 12) l-(2+0-difluorobenzoyl)-8-(
4-hydroxyphenyl)D7[;■8)L-(2+6
-difluorobenzoyl)-8-(4-schif-4-hexylphenyl))14 element+14)L-<2.6-difluorobenzoyl)-8-(4-)lifluoromethoxyphenyl)urea+15)L-(2 ,6-difluorobenzoyl) -3Iper-(8,4-dichlorophenyl]urea; 16)L-(2-lytetralobenzoyl)-3-(4-trifluoromethylphenyl) bj< element +17)l-
(2,6-dichlorobenzoyl)-3-(4-chlorophenyl)urea; 1B) L-(2,6-dichlorobenzoyl)-3-(4
-)Lifluoromethylphenyl)urea JL9) ■-benzoyl-3-(4-chlorophenyl)urea di20)L-(2,6-difluorobenzo-r)-3-
(2-hydroxy-4-trifluoromethylphenyl)
It was confirmed that urea; 21) l-(2-chlorobenzoyl)-3-(2-hydroxy-4-trifluoromethylphenyl)urea suppresses tumor tumor flHJ in some test models.

Eight test models were used to determine the inhibitory effect of the compound in vivo: 1,057BII/6 Metastatic tumors such as B-16 melanoma have the advantage of providing both the primary site and the microscopic lesions for studying the effects of drugs or other agents on cancer.

n, AKR 20-day mouse paste Tsujiway bone meat 11itj (
Rldge-way Osteogenic Sarc
The advantage of using such localized, non-metastatic tumors for studies of the efficacy of oma(RO8) drugs is that they allow for the perturbation of tumor growth and longevity in mice compared to IB versus primary tumors.
This point can be attributed to the contact effect.

IIl, AKR Mouse Skin Cancer (Ca 1025
) This tumor shows a secondary cell type that arises from the skin, and malignant melanoma is not malignant.

Compound tests were conducted by intraperitoneally injecting a suspension of the compound in polyvinylpyrrolidone aqueous solution. The dose and schedule of injections have been varied, but the best '72 schedule for any compound has not yet been determined.

Mortality rates for compounds were very low except for PH60-44. Injection of the compound is done with two hands [(to
Even doses as large as 0 mg were not seriously lethal.

PH (30-44 cases of mortality was seen with the drug jjf, which was much lower than this.
When given multiple injections with 9 levels, [0-15%
The mortality rate was Therefore, for each experiment, 10m9
8 injections or 6 injections of 41 mg each.

Testing of compounds followed the following procedure.

Two measures with malignant melanoma 1] Diflubenzuron 20m for mice
9 or 4 (l) of T4 (J suspension or suspension) were injected (blank experiment). In two of the experiments, 20 mg
Treatment (p) with the first injection of diflubenzuron reduced tumor size by 1[,7% and [3,4%] within 24 hours, whereas in the blank ivt:l-treated controls the tumor size decreased by 1[,7% and [3,4%]. The size of the tumor increased by 60.7% and 34%, respectively (Table [2009]). This effect was also due to the change in doubling time (i.e., the doubling of tumor size within 418 hours after treatment). The average time (expressed by the day of release) in the case of blank-treated controls increases significantly from day 9 to day LO04 in animals treated with diflubenzuron 1 (Table [2003]).

4 (H1+9 injection increased -lu1 efficacy (
22% initial decrease and negative doubling time).

Table [3 *In Tables [~[5], the number of tumors measured is shown in parentheses. Negative values indicate a decrease in tumor size.

A negative doubling time indicates the time to decrease by half the size.

In yet another experiment, 2°
9 injections continued to retard tumor growth (Table 2). After only one injection, tumor reduction remained for approximately 2 days. However, 20m on day 2 or 4
When q injections were repeated, the formation rate decreased until approximately the 6th El.

Table 2. Total of 1 for 3 or 5 injections within 6 days after the first injection.
．． Repeated injections of 00 m9 reduced tumor membership by day 11 (Table 3).

On days 2 and 19 (ul), tumor growth was visibly slower than controls, but not significantly. However, after day 19, treated tumors were in fact in +ffF arrest and on day 20
The increase in and 21 was smaller than the blank test laboratory 1 to 1 control.

Table 3 The same experiment (2) as was conducted with diflubenzuron was carried out at PH
60-411.

,, P HQ tl on day 0 for a 20-day-old mouse with melanoma.
The first injection of -441 caused a tumor count m11- for approximately 24 hours (Table 4). However, by administering the second and third injections to 11 and 5, respectively, most of the cases of subsequent ulcers will occur.)
There was no effect of sitting (Table 4). Tumor growth after 5 days or more was unaffected as evidenced by the percentage increase in tumor size.

Table 4 Similar experiments were also conducted using the compounds 2), 3), 5), 6) and [0) to 19).

Comparisons were made in flJ of treated tumors versus tgt*a with similar average size.

Compounds 8), 5), 6), to), [1), L2),
In the case of 1.5) and 19), administer 20TnQ each of mulberry f[j to [Io, 1.2.8 and 4, and compound L
In the cases of 8), [4ノ, 16), 17) and [8], only io my of Mulberry 1'a was administered on days 0, [, 2, 3 and 4]. . Compound 2) in Iυ bales
In [l011 and 2, a dose of 20m9 each was administered.

The results are summarized in Table 5.

, B, AKR Tsujiway bone and meat +1! -1'
i (ROS) of diflubenzuron in this test model;! 2omg on day 0 as H1j jH) (Table 6)
, 201119 on days 0 and 8; Father's Day OS l, 2.5
and 6 and 2 Q m9 each (Table 7) were used. Similar to the malignant melanoma model, a decrease in growth rate was seen 24-48 hours after the first injection of diflubenzuron (Table 6)
However, the size of the injury did not decrease.

Table 6: Repeated injections decreased the incidence of lung cancer, but the rate of lung cancer development did not decrease even after the number of treatments was exceeded (Table 7).

Table 7 This trash! In the tumor model, the effect was smaller (or treatment fKj was less effective) than in the black lung model.

Only one administration of PH80-4+4 reduced the growth rate of lilt tumors in 24 hours (Table 8). This table shows the fish size of 80m1) PH60-41.
The results of two experiments in which KR was injected into mice are shown. However, the schedule for injections is day 0. ．． 1 and 2, and the second on days 0.8 and 5. The results of the second experiment responded very well, with only an increase in tumor stasis or activity at 2F,1 and a decrease in the percentage increase over 11 days. In contrast, the delayed schedule (days 0.3 and 5) was no more effective than the first injection.

In this model, the initial effects of treatment with PH60-44 in both experiments were slightly greater than the initial effects of diflubenzuron in the same model.

Table 8. 0. ! KR Nisun January 2nd "ρ-pi" 1st (3
In this model, diflubenzuron n
Multiple injections of iH decreased tumor size within 24 UM and decreased growth rate within 48 hours (Table 9).

Table 9 However, multiple doses of mulberry did not significantly alter tumor growth more than the first injection of M.

Also, in this model, the compound 'I? lPH6o~41・
was also tested. For 20 days, 51 ng each 'fi: 'T + Muken Kr Klj or 0.05- of medium only and day 0
, [, 2, 3 and wound or L Om9 each corner suspension? (l or medium only 0.Lfnl is 1Elo,
T and 5 were treated.

The first injection of 5 or I In contrast, the blank treatment control increased by 54.4% on day 1 and 76.6% on day 2 (Table 10).

Multiple injections during the first 5 days after the first injection reduced tumor growth throughout the 2 day study period.

Table [O From the results of the above test model, it appears that the antitumor effect of diflubenzuron is probably drug-based schedule dependent. In malignant melanoma models, increasing the intensity and prolonging administration of diflubenzuron -1j- enhances and prolongs tumor control.

PH60-14 has a moderate effect on malignant melanoma tumors, with no tumor growth for 24+ +B7 after only one injection. However, PH60-+ 4+ is GaL025
A strong and subtle effect was produced in skin IN cancer.

A series of experiments were carried out to test the possibility that the metabolism of L-benzoyl-3-phenylurea, which has known insecticidal properties, has an anticancer effect. Do it! The effects of metabolic inhibitors and the intermediate Pi on the action of diflubenzuron in an animal model with unilateral black lung tumor were determined.

057 B L / 6 threads two hands day mouse txto Cao'
After injecting I11 melanoma I1Ill tumor cells, an Mj lagoon of approximately 50-500 m9 was formed at the injection site in 20-day-old mice after 14 days. Animals with growing tumors were treated with diflubenzuron [0O1l19(1'
(suspended in polyvinylpyrrolidone water bath solution) or with metabolic inhibitors or metabolic inducers.

I used cobaltous chloride as a metabolite once a month. This compound inhibits the monooxygenase action of liver 11fJli microsomes.

Femebarbital and 3-methyl- as metabolic inducers
Furanthrene was used.

Phenobarbital is a general compound (a zygosidase inducer that increases the metabolism of various substances). It greatly increases the action of biphenyl hand-hydroxylase, but
- Only slightly increases hydroxylase action.

3-Methylfuranthrene is an inducer of microsomal mixed-function oxidase action. This primarily serves to hydroxylate (oxidize) aromatic compounds. 3-Methylfuranthrene induces 2-hydroxylase action but not Φ-hydroxylation.

Treatment with diflubenzuron alone reduces tumor size in days 1-3. However, when the amount of primary cobalt was increased to 4 fJ of diflubenzuron, the inhibitory effect of diflubenzuron decreased (Table [2).

From this experiment, it can be concluded that the action of reducing the monooxygenase action of liver microsomes also reduces the anticancer action of diflubenzuron. These data support the hypothesis that enzymatic action in the liver chromosome produces a metabolite of diflubenzuron, which is a more active anticancer agent than the original compound.

Table 12E. If the effect of diflubenzuron on the metabolic inducer vI alr F is directly related to the action of diflubenzuron, then the anticancer effect of diflubenzuron by the enzyme 1 乍月 1 involved by the injection of mexidase 1 and pI. The effect can be increased.

In Sasuno's experiment, fenobarbital was used to treat liver ID'A.
il, '6, and thus liver microsomal r.
Increased 1r action. Phenobarbital 5: I
Rats were injected with diflubenzuron (Σso< before the desired treatment).

As shown in Table 13, fumebarbital greatly increases the action of biphenyl 4-hydroxylase and only slightly increases 2-hydroxylation, but the anticancer action of diflubenzuron is significantly increased. Met [i.

Table 13 Does increasing the 2-hydroxylation of diflubenzuron improve the anticancer properties of diflubenzuron 111? For the purpose of determining whether the
It was administered before i9H treatment. 8-Mo enhances 2-hydroxylase action but not mountain-hydroxylase action. As a control, a small group of tumor-bearing 20-day-old mice were injected with 8-M0m. 3-Mo enhances the anticancer effect of diflubenzuron by 5-El (7) 7111
(Table 14).

Table 14 Furthermore, as is clear from the table, 3-M0 alone does not inhibit tumor growth.

As an experiment to further prove the test results described in Sections 8 and B, a group of 20-day-old mice with lung cancer were given 20 m each of diflubenzuron 100 m9.3-hydroxy substitute product or 2-hydroxy metabolite [00 m9].
9 were administered on days 0, [, 2, 3 and 4].

The 2-hydroxyl dq1 product is processed 1r after the final 4
- More effective than the original compound, diflubenzuron, over a period of 8 hours (Table [5)]. Furthermore, the 3-human oxy metabolite had no agonizing effect.

Envy [5 These results fully support the data showing that the 2-hydroxy metabolite of diflubenzuron is more antitumor than diflubenzuron itself. [%do. The doubling time observed in cell culture for this type of cell is 12~
It is 161 hours. For this experiment, the surface door i 6 c
A 6-well tissue culture plate with an m2 flat bottom design appeared suitable and was used.

On day 0, an amount of melanoma B-16 cells was transferred to a cell culture plate.

The compounds to be tested were sonicated with 4311 L and then kept at 87°C for 3 hours without cells.

The test compound was then administered in the desired amount to the plates containing melanoma B-16 cells (each test was done in triplicate), after which the plates were incubated at 37°C for 20 hours in a CO□ constant-humidifier. Then, incubation was stopped, the culture medium containing 11 test compounds was removed, the cells were washed twice, and 1 L of fresh culture was added.

48 hours after the start of the culture phase, each plate''-, tllllllll
constant cell coulter counter (microcel, IC
oult, erQCounter). The results obtained in this manner were expressed as 1 and 0 as a percentage of controlled cell growth. The results are shown in Table 16.

Table 16 In the above experiments, test compounds were used in the form of KengJ solution. The solubility and strength of the active compound is not completely understood.
- Comparison of the above results is sufficient.

Therefore, some compounds were dissolved in ethanol.

In the same way as above, i.e. in the test on melanoma B-16 cells, r? ! The results are shown in Table 17.

Table 17 Ethanol itself has no effect on melanoma B-16 cell proliferation.

Claims (1)

[Scope of Claims] 1. L-benzoyl-3-phenylurea or a known metabolite thereof, which has an anticancer effect and an insecticidal effect of at least one persimmon, as an active ingredient, together with a pharmaceutically acceptable carrier. A pharmaceutical composition having an anticancer effect, characterized by having a horn. 2 l-benzoyl-8-phenylurea is compound 1) l-(2,6-difluorobenzoyl), 3-(4
-chlorophenyl)urea; 2) l-(2,6-difluorobenzoyl)-8-[8
,5-dichloro-4-(8-chloro-5-trifluoromethyl-2-pyrisiloxy)phenyl]urea; 8) L-(2,6-difluorobenzoyl)-8-[!
l-chloro4-(4-chlorophenoxy)phenyl]
Urea 1 4) L-(2,6-difluorobenzoyl)-8-(4
5) L-(2-chlorobenzoyl)-8-(4-chlorophenyl)urea 1 6) L-(2-chlorobenzoyl)-8-(4-trifluoromethoxyphenyl) urea! 7) L-(2,6-difluorobenzoyl)-8-(4-chloro-2-hydroxyphenyl)urea; 8) L-(2,6-difluorobenzoyl)-3-(4
-chloro4-8-hydroxyphenyl)urea; 9) l-(2-chlorobenzoyl)-a-(2-hydroxy-4-trifluoromethoxyphenyl)urea t 10) L-(216-difluorobenzoyl)-3 −(
4-cyanophenyl)urea; 11) L-(2,6-difluorobenzoyl)-ill
-[4-(Lt l*2+3+8+8-hexafluoropropoxy)phenyl]urine; 112) l-(2,6-
difluorobenzoyl)-3-(4- and do-4-doxyphenyl)urea + 13)l-(2,6-difluorobenzoyl)-8-(4-cyclohexylphenyl)urea 1 14)l-(2,6-difluorobenzoyl) -3-(
4-trifluoromethoxyphenyl)urea; 15) L-(216-difluorobenzoyl)-8-<
8.4-dichlorophenyl)urea; 16)' 1. −
(2-chlorobenzoyl) -8-(4-trifluoromethylphenyl) urine* +17)1. -(2,6-dichlorobenzoyl)-8-(4-chlorophenyl)urea; 1B) L-(2,6-dichlorobenzoyl) = 3-(4
-trifluoromethylphenyl)urea; 19) ■-benzoyl-8-(4-chlorophenyl)urea; 20) L-(2,6-difluorobenzoyl)-3-(
and 21) L-(2-chlorobenzoyl)-3-(2-hydroxy-4-trifluoromethylphenyl)urea t. - The composition according to claim 1.