Pharmaceutical compositions and methods for treatment and prophylaxis of cancer

A pharmaceutical composition comprising, in admixture with a pharmacologically acceptable carrier, an isomonool of the formula ##STR1## wherein R is an alkyl group with 1 to 5 carbon atoms, and n is an integer of 4 to 22, have carcinostatic, carcinostasis-reinforcing and carcinogenesis-preventing activity.

BACKGROUND OF THE INVENTION 
This invention relates to novel pharmaceutical compositions and methods for 
treatment and prophylaxis of cancer, more specifically for carcinostasis, 
reinforcement of carcinostasis and prevention of carcinogenesis. 
Up to now, several types of carcinostatic agents have been known. These 
include; alkylating agents which exhibit the carcinostatic activity by 
alkylating such biopolymers with indispensable roles as nucleic acids and 
enzymes, metabolic antagonists which inhibit metabolism of nucleic acids, 
mitotic poisons which affect biosynthesis of nucleic acids in the cell, 
carcinostatic antibiotics which exhibit cytocidal activity against the 
cells in rapid proliferation, carcinostatic compounds of plant origin, and 
hormones. 
On the other hand, amphotericin B and the like have been investigated as 
carcinostasis-reinforcing agents which may exhibit effects on improvement 
in absorption of drugs with poor absorbability to cancer cells and thus 
enhance carcinostatic effects of carcinostatic agents such as bleomycin 
and adriamycin. However none of them have come to attain any notable 
effect. 
Any of carcinostatic agents known from the prior art do not get rid of 
causing side effects, and few of them exhibit a satisfactory effect. 
Consequently, carcinostatic agents with improved characteristics have been 
needed. And there also have been needs for excellent 
carcinostasis-reinforcing agents in order to let carcinostatic agents 
express their full effects. 
Meanwhile, there is a leading hypothesis that carcinogenesis may be 
triggered through two steps mediated by an initiator and a promoter. 
However, the mechanisms of carcinogenesis is not known in relation to 
their influences exerted upon human body in daily life. Thus, prevention 
of carcinogenesis can only be pursued by a trial-and-error method based on 
experiences. Consequently, the development of carcinogenesis-preventing 
agents has been far behind that of anti-cancer agents. 
The criteria for carcinogenesis-preventing agents may be as follows. 
First, carcinogenesis-preventing agents, in general, are required to 
possess greater safety than that required for anti-cancer agents, and they 
have to be totally free of side effects. 
Secondly, they are required to be effective by oral administration so as to 
be administered easily in daily life. 
Thirdly, they are required not only to be effective on particular 
experimental cancers but also to have a wide spectrum against various 
types of cancers which have actually the higher rate of incidence. 
On the basis of aforementioned situation, the inventors have screened 
substances of sebum origin in order to find compounds which will be useful 
as agents for treatment and prophylaxis of cancer, assuming that the 
development of such agents should be started from the naturally and 
physiologically occurring "bio-substances" in the body of higher animals. 
As the result, some compounds that fulfill the aforementioned requirements 
were discovered. 
These compounds have both carcinostatic and carcinostasis-reinforcing 
effects. 
Moreover, they are thought to be useful for general prevention of 
carcinogenesis, for prevention of recurrence after completion of cancer 
treatment, for prophylaxis of occupational cancer and for prophylaxis in 
those who are hereditarily liable to certain cancers. Since the compounds 
of the present invention are chemically stable and can remain unchanged 
after cooking, they may be used as food additives and for functional foods 
or health foods. The compounds of the present invention are compounds 
which occur naturally in sebum, and they may, accordingly, be readily 
accepted by general people. 
SUMMARY OF THE INVENTION 
Through an intensive investigation for a carcinostatic, 
carcinostasis-reinforcing or carcinogenesis-preventing agent which would 
cause no side effects, the present inventors have discovered that 
isomonools represented by the general formula: 
##STR2## 
wherein R is an alkyl group with 1 to 5 carbon atoms, and n is an integer 
of 4 to 22, are excellent in such activities. 
Thus, the present invention provides carcinostatic, 
carcinostasis-reinforcing and carcinogenesis-preventing compositions 
containing as an active ingredient an isomonool represented by the general 
formula (I): 
##STR3## 
wherein R is an alkyl group with 1 to 5 carbon atoms, and n is an integer 
of 4 to 22. 
DETAILED DISCUSSION 
In the present invention, the length of the alkyl chain of the isomonools 
in the general formula (I) is defined by n which is an integer of 4 to 22 
and R which is an alkyl group with 1 to 5 carbon atoms. When n is less 
than 4, none of carcinostatic, carcinostasis-reinforcing and 
carcinogenesis-preventing activities are exhibited. Likewise, when n is 
greater than 22, none of such activities are exhibited. When R is an alkyl 
group with 6 or more carbon atoms, none of such activities are exhibited, 
either. A marked increase in potency in activities is observed when n is 
10 to 16, and when R is methyl or ethyl in the general formula (I). Among 
others, 14-methyl-1-pentadecanol of the formula (II): 
##STR4## 
and 14-methyl-1-hexadecanol of the formula (III): 
##STR5## 
are especially potent in activities and, therefore, particularly 
preferred. 
The isomonools utilized in the present invention may be produced by 
decomposing naturally occurring compounds such as waxes. Examples of such 
naturally occurring compounds include wool wax, spermaceti, beewax, white 
wax and carnauba wax. 
The isomonools utilized in the present invention may be obtained, for 
example, as follows (FIG. 1, a flowchart of the production process). 
SAPONIFICATION 
A naturally occurring compound such as wool grease is suspended in water in 
the presence of 1.18 times of an alkali (NaOH) on molar basis, and 
saponification reaction is conducted for 3 hours while stirring at 
135.degree..+-.5.degree. C. under pressure in an autoclave. 
SEATION OF ALCOHOLS FROM FATTY ACIDS 
To the resulting saponification product (the mixture of sodium salts of 
higher fatty acids and higher alcohols) are added water and 
methylethylketone (hereinafter referred to as MEK). The mixture is 
transferred into a separating funnel and heated to 70.degree. to 
75.degree. C. to extract alcohols into MEK. The resulting solution of wool 
alcohols in MEK is evaporated in vacuo to give wool alcohols as a solid 
matter. 
MOLECULAR DISTILLATION OF WOOL ALCOHOLS 
The solid wool alcohols thus obtained is subjected to molecular 
distillation to obtain a fraction with a lower boiling point (temperature; 
&lt;80.degree. C., pressure; 1.times.10.sup.-2 Torr). The fraction is 
hereinafter referred to as MD1-Alc. 
FRACTIONATION OF MD1-ALC BY REVERSE PHASE COLUMN CHROMATOGRAPHY 
MD1-Alc is separated into 6 fractions by reverse phase column 
chromatography (open column). 
FRACTIONATING CONDITIONS 
Solid phase: crushed ODS, pore size 60 .ANG., particle size 60/200 mesh 
(commercial name; YMC.GEL, YAMAMURA KAGAKU KENKYUSHO K.K.) 
Eluant: CHCl.sub.3 /CH.sub.3 OH/H.sub.2 O=5/15/1 (in volume). 
The third fraction, which has the third highest polarity, is hereinafter 
referred to as ODS#3. 
FRACTIONATION OF ODS#3 BY NORMAL PHASE COLUMN CHROMATOGRAPHY 
ODS#3 is fractionated by normal phase column chromatography (open column). 
FRACTIONATING CONDITIONS 
Solid phase: silica gel, pore size 60 .ANG., TYPE 60 .ANG. SPECIAL, 
particle size 100/200 mesh (commercial name; SILICAR (trademark): produced 
by Mallinckrodt, distributed by DAIICHI KAGAKU YAKUHIN K.K.) 
Eluants: 
Eluant I, CHCl.sub.3 /CH.sub.3 COCH.sub.3 =96/4 (in volume) 
Eluant II, CH.sub.3 OH. 
The first and the second fractions are eluted with the eluant I, and the 
third with the eluant II. The second eluted fraction is hereinafter 
referred to as ODS#3-2. 
FRACTIONATION OF ODS#3-2 BY HPLC 
Separation and isolation of isomonools 
ODS#3-2 is fractionated by HPLC to isolate the aimed compounds. 
HPLC CONDITIONS 
Column: TSK gel ODS-120T (commercial name, TOSO K.K.), 21.5 mm ID.times.300 
mm 
Mobile phase: CH.sub.3 OH/H.sub.2 O=95/5 (in volume) 
Flow rate: 5.0 ml/min 
Column temperature: room temperature. 
The isomonools may be separated and isolated also by other methods. 
Examples of such other methods include the methods described by SATOSHI 
TAKANO, MAKOTO YAMANAKA, KIKUHIKO OKAMOTO and FUMIO SAITO (Allergens of 
lanolin: parts I and II, Part I: ISOLATION AND IDENTIFICATION OF THE 
ALLERGENS OF HYDROGENATED LANOLIN, JOURNAL OF THE SOCIETY OF COSMETIC 
CHEMISTS, 34, P 99-116 (1983)). 
Examples of isomonools include iso--C.sub.14 --OH [12-methyl-1-tridecanol], 
iso--C.sub.15 --OH [13-methyl-1-tetradecanol], iso--C.sub.16 --OH 
[14-methyl-1-pentadecanol], iso--C.sub.17 --OH [15-methyl-1-hexadecanol] 
and iso--C.sub.18 --OH [16-methyl-1-heptadecanol] when R is methyl, and 
iso--C.sub.15 --OH [12-methyl-1-tetradecanol], iso--C.sub.16 --OH 
[13-methyl-1-pentadecanol], iso--C.sub.17 --OH [14-methyl-1-hexadecanol], 
iso--C.sub.18 --OH [15-methyl-1-heptadecanol] and iso--C.sub.19 --OH 
[16-methyl-1-octadecanol] when R is ethyl. 
These isomonools may be used alone or in mixture with one or more other 
isomonools. 
For preparing a composition of the present invention into the forms of 
injection or instillation, the isomonools may be admixed with a surfactant 
such as Pluronic F-68 (commercial name of a poloxamer, ASAHI DENKA K.K.), 
HCO-60 (commercial name, NIKKO CHEMICALS K.K.) and the like, and then 
dispersed with the aid of ultrasonic waves, or the isomonools may be 
processed into a type of composition such as a liposome suspension or an 
oil-in-water emulsion. 
Such compositions may contain preservatives such as methyl 
p-hydroxybenzoate, stabilizers such as lecithin or linoleic acid, 
non-aqueous vehicles such as coconut oil and agents for suspension aid 
such as glucose. 
For preparing composition for oral administration, the isomonools may be 
made into the form of capsules suitable for intestinal absorption by 
incorporating, for example, binders such as gelatin, stabilizers such as 
magnesium stearate, diluent bases such as lactose and disintegrators such 
as potato starch and by coating the capsules with acetylphthalylcellulose 
or methyl acrylate/methacrylate copolymer to form an enteric coating 
layer. 
The isomonools may also be made into the forms of granules, 
sustained-release capsules for implant, a suppository, a nebulizer or a 
buccal preparation. 
For use as a carcinostatic composition or a carcinogenesis-preventing 
composition, the dose (per 1 kg body weight per day for adult human) of 
the active ingredient may be preferably 5 to 1200 mg, particularly 
preferably 20 to 300 mg for parenteral preparations such as for 
intravenous or subcutaneous injection or instillation, and preferably 0.1 
to 40 g, particularly preferably 0.5 to 8 g for oral preparations, for 
example, capsules. 
For use as a carcinostasis-reinforcing composition, the dose may be 
preferably 0.45 to 102 mg, particularly preferably 1 to 28 mg for 
parenteral preparation, and preferably 0.005 to 5.2 g, particularly 
preferably 0.07 to 1.2 g for oral preparations. 
The agents of the present invention are effective in treatment not only of 
ascites tumors and leukemia but also of solid tumors which include 
adenocarcinoma, squamous cell carcinoma, undifferentiated carcinoma and 
sarcoma in various tissues. The agents are effective not only in 
tumor-implanted animals but also on cultured malignant cells from human, 
mouse, rat, hamster and so on. It is thus indicated that the agents may 
have a direct lethal effect on tumor cells without specificity to animal 
species, and, therefore, they may be used as chemotherapeutic agents for 
cancer of human or domestic and other animals. 
In addition, they are effective not only by direct application to the 
implanted tumor site but also by application to a remote site. As for 
toxicity, LD.sub.50 is 5.1 to 17 g/kg for the rat by subcutaneous 
injection, and no side effects have been observed by repeated 
administration for 10 days at a dose of 1 to 2 g/kg.

The following is an example of the preparation methods of the isomonools, 
the active ingredients of the carcinostatic, carcinostasis-reinforcing and 
carcinogenesis-preventing compositions of the present invention. The scope 
of the present invention, however, is not limited by the example. 
EXAMPLE 1 
200 g of wool alcohol obtained by saponification of wool grease was 
subjected to molecular distillation to give 15.2 g of MD1-Alc as a 
fraction with a lower boiling point (temperature: &lt;80.degree. C., 
pressure; 1.times.10.sup.-2 Torr). 
The MD1-Alc was separated into 6 fractions by reverse phase column 
chromatography (open column) using CHCl.sub.3 /CH.sub.3 OH/H.sub.2 
O=5/15/1 (in volume) mixture as an eluant. The third eluted fraction 
(ODS#3) was obtained in an amount of 1.68 g. 
The ODS#3 was subjected to normal phase column chromatography (open column) 
and separated into 3 fractions using the eluants (the eluant I: CHCl.sub.3 
/CH.sub.3 COCH.sub.3 =96/4 (in volume), the eluant II: CH.sub.3 OH). 
The second eluted fraction (ODS#3-2) of the three was obtained in an amount 
of 0.96 g. 
ODS#3-2 was fractionated by HPLC to isolate each of the aimed compounds. 
Thus obtained were 14-methyl-1-pentadecanol, 14-methyl-1-hexadecanol, 
16-methyl-1-heptadecanol, 16-methyl-1-octadecanol, 
18-methyl-1-nonadecanol. 
Capillary gas chromatography clearly demonstrated that each of these 
isomonools was isolated. 
For determining chemical structures, .sup.13 C-NMR and GC-MS were applied, 
and these isolated compounds were identified to be the isomonools 
represented by the general formula (I) of the present invention. 
FIG. 2 shows a .sup.13 C-NMR spectrum (100.40 MHz, CDCl.sub.3, .delta. ppm) 
of 14-methyl-1-pentadecanol, and FIG. 3 and FIG. 4 show GC-MS data 
thereof. FIG. 5 shows a .sup.13 C-NMR spectrum (100.40 MHz, CDCl.sub.3, 
.delta. ppm) of 14-methyl-1-hexadecanol, and FIG. 6 and FIG. 7 show GC-MS 
data thereof. Data shown in FIG. 3 and FIG. 6 were obtained in mode EI 
with an ionizing voltage of 70 eV and with the temperature of the ion 
source of 250.degree. C. Data shown in FIG. 4 and FIG. 7 were obtained in 
mode CI using isobutane as reacting gas, with an ionizing voltage of 200 
eV and with the temperature of the ion source of 250.degree. C. 
The isomonools obtained above were tested for carcinostatic, 
carcinostasis-reinforcing activity. 
TEST 1 
5-week old ddY mice were intraperitoneally inoculated with 10.sup.6 
Ehrlich's ascites carcinoma cells. Since 24 hours later, 50 mg/ml 
suspension of a test compound in 0.25% (% by weight. The definition is to 
be followed hereinafter) solution of Pluronic F68 in physiological saline 
was intraperitoneally injected to 10 animals/group at a dose of 10 
mg/kg/day for 5 days. 
The mean survival time after inoculation was 14.0 days for the group 
administered the carrier solution containing none of the test compounds. 
On the other hand, the mean survival time of the animal group administered 
the test compounds was 60 days or more for 14-methyl-1-pentadecanol, 57.5 
days for 14-methyl-1-hexadecanol, and 60 days or more for their 1:1 
mixture. The effects on prolonging the survival time were statistically 
significant. 
TEST 2 
6-week old F.sub.1 mongrel mice of C57BL/6 and DBA/2 were inoculated under 
back skin with 10.sup.6 adenocarcinoma 755 cells. Since 24 hours later, 50 
mg/ml suspension of a test compound in 0.25% HCO-60 solution in 
physiological saline was subcutaneously injected to 8 animals/group at a 
dose of 10 mg/kg/day for 5 days. The animals were sacrificed 10 days after 
the inoculation with adenocarcinoma 755 cells, and the tumor was excised. 
The mean tumor weight (g) of the control group was 6.5. On the other hand, 
that was 1.6 and 2.3 for the group administered 16-methyl-1-heptadecanol 
and 16-methyl-1-octadecanol, respectively. The tumor growth suppressing 
effects were statistically significant. 
TEST 3 
14-methyl-1-pentadecanol and 18-methyl-1-nonadecanol were tested for tumor 
growth suppressing effects on three different tumor cell strains 
(Ehrlich's mouse ascites carcinoma cells, human lung carcinoma A549 cells 
and mouse neuroblastoma NAs-1 cells). As the results, the two isomonools 
were demonstrated to be effective on any of these three types of tumor 
cells. 
These isomonools suppressed the colony formation rate of human lung A549 
cells to 2.2.times.10.sup.-3 and 4.1.times.10.sup.-3, respectively, by 
6-hour treatment at 10 .mu.M. 
These isomonools suppressed the growth of Ehrlich's ascites carcinoma cells 
to 4.5.times.10.sup.-3 and 1.5.times.10.sup.-2, respectively, by 5-day 
treatment at 10 .mu.M. 
The growth of mouse neuroblastoma NAs-1 cells was suppressed to 
7.8.times.10.sup.-3 and 6.5.times.10.sup.-3, respectively. 
TESTS 4 AND 5, AND COMING TEST 1 
The cytocidal activity (IC.sub.90) of bleomycin was evaluated when used 
alone or used in combination with 14-methyl-1-pentadecanol or 
14-methyl-1-hexadecanol, using Ehrlich's mouse ascites carcinoma cells and 
human lung tumor A549 cells. The results are shown in Table 1. 
As clearly shown from the results in Table 1, when the isomonools of the 
present invention is used in combination with a carcinostatic agent, the 
cytocidal effect is markedly increased in comparison with the case where 
the carcinostatic agent is used alone. 
TEST 6 AND COMING TEST 2 
The amount of bleomycin taken up into Ehrlich's ascites cells was 
determined as a function of time in the case where bleomycin (60 .mu.g/ml) 
was used in combination with 14-methyl-1-pentadecanol (0.05 .mu.M). The 
results are shown in Table 2. 
As clearly shown from the result in Table 2, when the isomonool of the 
present invention is used in combination with a carcinostatic agent, the 
uptake of the carcinostatic agent into Ehrlich's cells is rapidly 
increased in comparison with the case where the carcinostatic agent is 
used alone. 
TEST 7 AND COMING TESTS 3 AND 4 
5-week old male C57BL/6 mice were intraperitoneally inoculated with 
10.sup.6 Ehrlich's mouse ascites carcinoma cells. Since 24 hours later, a 
50 .mu.g/ml solution of bleomycin in 0.25% HCO-60 solution in 
physiological saline, and a suspension containing 50 .mu.g/ml of bleomycin 
and 5 .mu.g/ml of 14-methyl-1-pentadecanol were intraperitoneally injected 
at a dose of 10 mg/kg.multidot.day and 11 mg/kg.multidot.day, 
respectively, to each 8 animals/group for 5 days. The survival time of the 
injected mice was then followed. The results are shown in table 3. 
As shown in Table 3, the mean survival time after inoculation was 33.3 days 
for the group (Comparing test 3) which was administered the bleomycin 
solution containing none of the carcinostasis-reinforcing agents of the 
present invention, whereas the mean survival time after inoculation was 
54.3 days for the group (Test 7) which was administered the bleomycin 
solution containing one of the isomonools as a carcinostasis-reinforcing 
agent of the present invention. 
Thus, it is demonstrated that the isomonools of the present invention are 
useful as carcinostasis-reinforcing agents. 
TEST 8 
6-week old female SD rats (n=90) were treated as follows using 
14-methyl-1-pentadecanol as a carcinogenesis-preventing agent. 
Unadministered group: n=30. MF powder feed (ORIENTAL KOBO K.K.) was given 
as a basic feed. 
Administered group A: n=30. A test feed was given continually (for 7 days). 
The feed was prepared by admixing the said carcinogenesis-preventing agent 
(after emulsified in Pluronic F-68) with the abovementioned basic feed in 
an amount of 50 mg/kg. 
Administered group B: n=30. A test feed was given continually (for 7 days). 
The feed was prepared by admixing the said carcinogenesis-preventing agent 
(after emulsified in Pluronic F-68) with the abovementioned basic feed in 
an amount of 250 mg/kg. 
Among these groups, there were no significant differences in the amount of 
feed consumed. 
After the aforementioned treatment, 5 mg/body of benzanthracene (DMBA: a 
carcinogenic compound) was administered by injecting subcutaneously in 
mammary gland 0.1 ml/body of a 50 mg/ml benzanthracene solution. Then, 
each of the aforementioned groups was given corresponding feed for 20 
weeks. 
Among these groups, there were no significant differences in the amount of 
feed consumed. 
After the completion of the treatment, the rats were sacrificed and 
observed for the occurrence of mammary cancer. The results are shown in 
Table 4. 
TEST 9 
40 male Wistar rats (8-9-week old, body weight; 110 g) were given a 
drinking water containing 167 .mu.g/ml of 
N-methyl-N'-nitro-N-nitrosoguanidine (MNNG: a carcinogenic compound) and a 
feed (dry pellets CE-2, produced by CLEA JAPAN) for 210 days. 
On the 210th day, adenoma-like hyperplasia was observed in the stomach. 
The rats were allotted to 5 groups and treated as follows since the 211th 
day. 
Unadministered group: n=8. 
Administered group A: n=8. The compound A of the present invention (See 
below. The definition is to be followed hereinafter) was administered 
orally in an amount of 250 mg/kg feed for 126 consecutive days. 
Administered group B: n=8. The compound B of the present invention was 
administered orally in an amount of 250 mg/kg feed for 126 consecutive 
days. 
Administered group C: n=8. The compound C of the present invention was 
administered orally in an amount of 250 mg/kg feed for 126 consecutive 
days. 
Administered group D: n=8. The compound D of the present invention was 
administered orally in an amount of 250 mg/kg feed for 126 consecutive 
days. 
A: 14-methyl-1-pentadecanol 
B: 14-methyl-1-hexadecanol 
C: 16-methyl-1-heptadecanol 
D: 16-methyl-1-octadecanol. 
On the 337th day, the survining rats were sacrificed and the stomachs were 
(1) subjected to Borrmann's classification according to gross observation 
and (2) histologically observed (haematoxylin-eosin stain or ASAN stain), 
and then classified as follows. The results are shown in Table 5. 
Grade O: No stomach carcinoma are observed. 
Grade I: Stomach carcinoma within mucous membrane. 
Grade II: Stomach carcinoma infiltrating beneath mucous membrane. 
Grade III: Stomach carcinoma infiltrating into muscle layers or serous 
membrane. 
Grade IV: Stomach carcinoma metastasizing to adjacent lymph nodes, duodenum 
or jejunum. 
TEST 10 
5 groups of 6-week old male F344 rats were givn 0.05% (by weight. The 
definition is to be followed hereinafter) aqueous solution of 
N-butyl-N-(4-hydroxybutyl)nitrosoamine (BBN: bladder carcinogenic 
compound), as an initiator, by water supplying bottles for 4 weeks. Then 
the animals were given 5% sodium erythorbate aqueous solution, as a 
promoter (carcinogenesis promoting compound), and a feed which was 
prepared by admixing one of the compounds A-D of the present invention 
(See below) with the MF powder feed produced by ORIENTAL KOBO K.K. in an 
amount of 250 mg/kg. 
A: 14-methyl-1-heptadecanol 
B: 14-methyl-1-hexadecanol 
C: 16-methyl-1-heptadecanol 
D: 18-methyl-1-nonadecanol. 
Among the animal groups, there were no significant differences in the 
amount of feed consumed. 
After these treatment is completed, the rats were sacrificed and observed 
for the occurrence of pathological changes in bladder mucous membrane. The 
results are shown in Table 6. 
The carcinostatic, carcinostasis-reinforcing and carcinogenesis-preventing 
compositions of the present invention exhibit excellent effects, whereas 
the active ingredients thereof are obtained from organisms (higer animals) 
and, thus, may not cause severe side effects on organisms. 
TABLE 1 
__________________________________________________________________________ 
Cytocidal activity (IC90 (.mu.g/ml)) 
Carcinostasis 
Ehrlich's mouse 
Human lung 
Carcinostatic 
reinforcing agent 
ascites carcinoma 
carcinoma A549 
Test agent (0.05 .mu.M) 
cells cells 
__________________________________________________________________________ 
4 bleomycin 
iso-C.sub.16 --OH* 
4.0 28 
5 bleomycin 
iso-C.sub.17 --OH*.sup.2 
6.0 24 
Comparing T.1 
bleomycin 
-- 20 44 
__________________________________________________________________________ 
*.sup.1 14methyl-1-pentadecanol 
*.sup.2 14methyl-1-hexadecanol 
TABLE 2 
__________________________________________________________________________ 
Bleomycin uptake with lapse of time 
Carcinostatic 
Carcinostasis 
(.mu.g/mg Ehrlich's cell) 
Test agent reinforcing agent 
Initial 
30 60 120 
180 (min) 
__________________________________________________________________________ 
6 bleomycin 
iso-C.sub.16 --OH*.sup.1 
0 0.13 
0.26 
0.39 
0.55 
Comparing, 2 
bleomycin 
-- 0 0.03 
0.06 
0.12 
0.13 
__________________________________________________________________________ 
*.sup.1 14methyl-1-pentadecanol 
TABLE 3 
__________________________________________________________________________ 
Carcinostasis 
Number of surviving mice 
Carcinostatic 
reinforcing 
with the lapse of time (day) 
Test agent agent 0 14 
15 
16 
17 
18 
19 
20 
23 
33 
41 
45 
52 
57 
60 
__________________________________________________________________________ 
7 bleomycin 
iso-C.sub.16 --OH*.sup.1 
8 8 8 8 8 8 8 8 8 8 8 6 4 4 4 
Comparing, 3 
bleomycin 
-- 8 7 6 6 6 6 6 6 4 3 2 2 2 1 1 
Comparing, 4 
-- 8 8 6 5 4 3 1 0 0 0 0 0 0 0 0 
__________________________________________________________________________ 
*.sup.1 14methyl-1-pentadecanol 
TABLE 4 
______________________________________ 
Number of animals with mammary 
Group cancer/Number of tested animals 
______________________________________ 
Administered group A 
8/30 
Administered group B 
4/30 
Unadministered group 
15/30 
______________________________________ 
TABLE 5 
__________________________________________________________________________ 
Survival 
Borrmann's 
Histological 
Group (animal No.) 
Cause of death 
time (day) 
Classification 
classification 
__________________________________________________________________________ 
Administered 
(1) 
sacrificed 
337 I I 
group A 
(2) 
" " 0 0 
(3) 
" " 0 0 
(4) 
" " 0 0 
(5) 
" " 0 0 
(6) 
" " I I 
(7) 
" " 0 0 
(8) 
" " 0 0 
Administered 
(1) 
" " I I 
group B 
(2) 
" " 0 0 
(3) 
" " 0 0 
(4) 
" " 0 0 
(5) 
" " 0 0 
(6) 
" " 0 0 
(7) 
" " I I 
(8) 
" " I II 
Administered 
(1) 
dead in cage 
305 II III 
group C 
(2) 
sacrificed 
337 0 0 
(3) 
" " I I-II 
(4) 
" " 0 0 
(5) 
" " 0 0 
(6) 
" " 0 0 
(7) 
" " 0 0 
(8) 
" " II II 
Administered 
(1) 
dead in cage 
301 III III 
group D 
(2) 
" 312 III III 
(3) 
sacrificed 
337 I I 
(4) 
" " 0 0 
(5) 
" " 0 0 
(6) 
" " I I-II 
(7) 
" " I I-II 
(8) 
" " I II 
Unadminister- 
(1) 
dead in cage 
238 IV IV 
ed group 
(2) 
" 270 IV IV 
(3) 
" 270 IV IV 
(4) 
" 273 IV IV 
(5) 
" 280 IV IV 
(6) 
" 281 IV IV 
(7) 
" 285 IV IV 
(8) 
sacrificed 
337 III III 
__________________________________________________________________________ 
TABLE 6 
__________________________________________________________________________ 
Number of 
Number of animals with pathological changes in 
observed 
bladder mucous membrane (incidence, %) 
Group animals 
Papillary or tuberous hyperplasia 
Papilloma 
Cancer 
__________________________________________________________________________ 
Adminstered 
A 28 0 (0) 0 (0) 0 (0) 
group B 28 0 (0) 0 (0) 0 (0) 
C 28 1 (4) 0 (0) 0 (0) 
D 28 2 (7) 0 (0) 0 (0) 
Unadministered 
27 27 (100) 20 
(74) 
19 
(70) 
group 
__________________________________________________________________________