Note: Descriptions are shown in the official language in which they were submitted.
~)36618
The present invention relates to the novel substance,
tris(2-hydroxyethyl)ammonium-ortho-cresoxyacetate its prepa-
ration and pharmaceutical preparations containing the same.
The novel substance, tris(2-hydroxyethyl)ammonium-
ortho-cresoxyacetate, has the following formula:
[o-CH3C6H4OCH2COOl [NH(CH2CH2 )3~ .
The novel substance of this invention exhibits
antitumorigenic activity and as such is employed in medicine
as the active principle of a medicinal preparation for treating
malignant neoplasms.
The proposed substance is a white crystalline powder,
soft to the touch, devoid of smell, having a melting point
of 80 to 82C, with a pungent bitter-turning-sweet taste,
readily soluble in water and ethanol, and slightly soluble
in ether, benzene and carbon tetrachloride.
The antitumorigenic activity of the proposed
preparation was tested on spontaneous and induced tumors (strains
of sarcoma 37, 45 and 180, Pliss lymphosarcoma; Guérin-Passy
carcinoma; Brown-Pearce tumor; Walker carcinoma; and cancer
of the hepatic mucosa). 120 non-pedigree white mice, 400
albino rats and 24 rabbits were used in the experiment. Some
of the animals were allowed to live a natural span, a sure
indication of the practically total resolution of the tumors
and metastases (Brown-Pearce tumor and spontaneous tumors).
The preparation was admistered per os for 10 days
starting 48 hours after the inoculation. The degree of
suppression of tumor growth was calculated in percentage as
compared with the control group of animals.
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The activity of the proposed preparation is illustrated
by the following table.
Tumor suppression in ~ caused by the
following doses, mg/kg
Strain
100 200 300 400 600 800
mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg
. . .
Sarcoma 37 43 58 91 98
Sarcoma 45 80 90
Sarcoma 180 51 64 86 92 95 98
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Guerin carcinoma 51 60
Pliss lymphosarcoma 90 95
Walker carcinoma 95 98
Hepatic mucosa cancer 9o
The most effective therapeutic doses of the prepara- --
tion were found to be 300 to 400 mg/kg. At these doses, all ~ ,
the tumors studied were suppressed in their growth by 86 to 98
percent (except Guérin-Passy carcinoma which is suppressed in
its growth by more than 50 percent).
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The therapeutic effectiveness of the preparation was ~-
tested on spontaneous tumors of mice and induced Pliss lympho- :
sarcoma (rats) and Brown-Pearce tumor (rabbits). The course
of treatment was initiated 10 to 20 days after the strain
inoculation, during a period of intensive malignant growth. ;
The preparation in a dose of 400 mg/kg was administered per
os daily for 25 to 45 days. The results obtained are given
in the following table.
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Duration of the Number of cured
Animal speeies Type of tumor course ofanimals,~
treatment, days
i
Mice Spontaneous
tumor 30-40 75
Rats Pliss lympho-
sarcoma 25-35 100
Rabbits Brown-Pearce
tumor 45 100
The preparation exhibits a low level of toxicity. The
maximum toleranee dose of the preparation is 2,600 mg/kg;
LD50= 2,800 mg/kg; LDloo= 3,600 mg/kg (for white mice, per os).
In accordanee with the invention, the process for the
manufacture of said compound comprises reacting triethanolamine
with o-cresoxyacetic acid at a temperature of up to 100 C.
and subsequently recovering the desired product.
It is preferred that the process be effected in a
water-miscible organic solvent, preferably ethanol, acetone
or dioxane. The process is preferably carried out at a
temperature of from 50 to 80 C. In order that the desired
product may be pure enough for medical applieations, the
proeess is earried out in ethanol, and the desired product
is reeovered by the addition of diethyl ether to the reaction
mixture cooled down to -20 or -30C.
The proeess proeeeds as follows:
( 2CH2H)3 + o-cH3c6H4ocH2cooH -~
[ 3 6H4~CH2COO] - [NH(CH2cH2OH)3]+-
Triethanolamine is dissolved in a water-miscible
organic solvent, e.g. ethanol, acetone, dioxane, etc., mixed
103661~
with ortho-cresoxyacetic acid and heated to the point of
boiling. The process may be effected in the absence of a
solvent. In such a case the mixture of the parent components
is melted down (m.p. 78 to 83C).
The desired product is recovered by cooling the
reaction mixture and recrystallizing same. In order to obtain
a high-purity desired product fit for medical applications, the
process is effected in ethanol, and the desired product is
recovered by adding diethyl ether to the reaction mixture
cooled down to -20 or -30C. The yield of the desired product
constitutes up to 98 percent by weight of the theoretical. ~-
Practice of the novel process of this invention may be `
further understood by reference to the following examples of
manufacturing tris(2-hydroxyethyl)ammonium-ortho-cresoxyacetate.
Example 1
A solution of 149.2g (l.OM) of triethanolamine in
150 ml of ethanol is added to a solution of 174.5 g (1.05 M)
of o-cresoxyacetic acid in 500 ml of ethanolO The reaction
mixture is heated to the point of boiling, then cooled to -
a temperature of -20 or -30C, and 100 to 150 ml of diethyl
ether is added thereto. The white crystalline precipitate
is sucked off, washed with ether and dried in a vacuum. The
; yield of the desired product amounts to 295 g (93.6 percent
by weight); m.p. 82 to 83C. An additional 2.5 g of the
desired product is recovered from the mother liquor by
concentration in a hot-water bath.
Example 2 ;
149.2 g (l.OM) of triethanolamine is added to 166.2 g
(l.OM) of o-cresoxyacetic acid. As the reaction mixture is
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agitated, intensive, evolution of heat takes place. The
resultant viscous desired product is melted down (m.p. 78 to
80C) and rapidly cooled. The crystals are washed with ether
and dried. The yield is 315 to 317 g (100 percent by weight)
of the desired product with m.p. 80 to 81C.
Actual composition, wt.%: C, 58.83; 57.14; H, 8.18;
8.20; N, 4.46; 4.26. C15H25NO6
Estimated composition, wt.%: C, 57.12; H,8.00; N,4.44.
The present invention also provides a novel medicinal
preparation for treating malignant neoplasms.
In accordance with the invention, the medicinal
preparation comprises an active principle, tris(2-hydroxyethyl)
ammonium-orthocresoxyacetate of the following formula:
[ 3 6 4OCH2COOl ~H(CH2cH2oH)3~+
or said active principle combined with a pharmaceutical
solvent for injections or a pharmaceutical filler for tablets.
The preparation has a marked antineoplastic effect
and can be employed for the treatment of cancer of the stomach, -~
of the rectum, of the mammary gland, of the lungs, and of other
localizations.
The preparation has a peculiar effect on neoplastic
tissues: there appear therein collagenous fibres which
rapidly grow in number so that, with the neoplastic tissue
intergrown with connective tissue, its cells no longer
receive nutrition, cease dividing and degenerate.
The preparation of this invention is practically atoxic
and fails to suppress the growth of normal tissues having a high
level of proliferative activity, which favourably distinguishes -
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it from the currently used antineoplastic preparations.
The preparation was studied for acute toxicity
on white mice, albino rats and rabbits, weighing 17 to 20 g,
150 to 200 g, and 2 to 2.5 kg, respectively. In all animal
groups, the breakdown by sex was fifty-fifty. The animals
were kept under normal conditions and last fed 24 hours
before the experiment. The mice and rats were kept in the
laboratory for the entire observation period, whereas the
rabbits were returned, after 6 hours in the laboratory, to
the vivarium where they were observed twice a day. The
animals were fed for the first time six hours after the ~
preparation had been administered, subsequently they were -
put on a normal feeding routine. The preparation was
administered in three ways: orally, intraperitoneally and ;
intravenously. The effect of a single dose of the prepara- ~
tion was assessed according to Berence on 48 non-pedigree -
white mice. It was found that LDlo~=2,800 mg/kg and LD50= -~
2,600 mg/kg. The maximum tolerance dose was determined by ~ -
selection on 50 white mice to be equal to 2,400 mg/kg 40
albino rats received the preparation intraperitoneally
according to Kerber: LDloo=2,400 mg/kg; LD50=2,000 mg/kg-
; Intravenous administration tests were carried out on white
mice with the following results: at doses from 200 to 1,250
mg/kg, all the animals survived; LDloo--1,500 mg/kg;
LD50=1,300 mg/kg; the maximum tolerance dose = 1,250 mg/kg.
For rats which received the preparation orally according to
Kerber, LDloo=5,000 mg/kg; LD50=4,500 mg/kg; the maximum
tolerance dose = 4,000 mg/kg. For rabbits which received
the preparation intravenously, LDloo=1,500 mg/kg i
LD50~ 1,250 mg/kg; the maximum tolerance dose = 1,000 mg/kg
The clinical picture of the effect of the preparation
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administered orally in a maximum tolerance dose of 2,400
mg/kg developed in the following way: after the administration
of the preparation the latency period lasted for 5 to 20
minutes, testifying to the rapidity of its inhibition, after
which a marked reduction in motor activity was observed which
lasted for 8 to 20 hours. A day later the behaviour patterns
of the animals showed no abnormality. In doses of 800 mg/kg
and less, the preparation had no pronounced clinical manifesta-
tions. The antineoplastic activity of the preparation was
studied on induced and spontaneous tumours in mice, rats and
rabbits. The preparation was administered orally. Each
experimental group was made up of 6 to 12 animals. The
experiments were repeated two or three times. In determining
the suppressive effect of the preparation on tumour growth, -;
i~ was administered 48 hours after induction and then for 10
straight days. The inhibition process was evaluated by the
results of tumour growth in the control and experimental
animals. The results are given in Table 1.
'
No. of Tumour Animal No. of No. of Percentage suppres-
series strain species animals experi- sion
in the ments
group100 200 300 400 600 800
mg/kg
1.Sarcoma- Mice 10 2 51 64 86 92 95 99
180
2.Sarcoma- Mice 10 2 43 58 91 98
37
3.Sarcoma- Rats 10 3 80 90
4. Guerin Rats 10 3 51 60
sarcoma
5. LIO-l Rats 10 3 90 95
6. Walker Rats 10 3 95 98
carcinosar-
coma
7. Cancer of Rats 10 3 90
the hepatic ~
mucosa ~-'
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~)36618
In the course of treatment the general health status
of the animals improved; they gained 5 to 7 percent of
weight. The percentage suppression of tumour growth caused
by 300 to 400 mg/kg was 80 to 98, except for the Guerin
tumour which was suppressed by only 50 to 60 percent.
The therapeutic effectiveness of the preparation
was studied on the following tumorigenic strains: Sarcoma-45,
LIO-l, Walker carcinosarcoma, cancer of the hepatic mucosa in
rats, and Brown-Pearce tumour in rabbits. The treatment was
initiated 10 to 18 days after tumour induction, at the time
of its intensive growth. The control animals with induced
tumours underwent no treatment. The preparation was
administered per os in daily doses of 400 mg/kg until total reso-
lution of the tumour in most animals of the group occurred. At
regular intervals during the course of treatment individual
animals were butchered. The tumours were subjected to
histological examination during the treatment. Blood samples
were taken from rabbits with Brown-Pearce tumours. In the
control groups with induced tumours, the animals lost weight
in spite of the growing large tumours. Spontaneous resolution
~ was not observed. All the animals of the control groups died
-~ within 20 to 35 days. The results of the treatment are given
- in Table 2.
.,
No. of Tumour Animal No. of Duration Wt. Wt. % of
series strain species animals of treat- prior after ani-
ment, days to treat- mals
treat- ment,g with
ment, g resolved
tumours
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1. Sponta- Mice 8 35 27 32 75
neous
tumour
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2. Sarcoma- Mice 10 30 25 30 100 ~ -
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3. Sarcoma- Rats 10 35 190 210 100
4. LIO-45 Rats 10 30 230 242 100
5.Brown- Rabbits 12 45 2,550 2,620 100
Pearce
tumour
The tumours were completely resolved in all animals
within 30 to 45 days. In the follow-up lasting for 2 to 4
months, no relapses were observed, as confirmed histologically.
The blood picture of the rabbits with Brown-Pearce
sarcoma treated with the proposed preparation is given in
Table 3.
Blood counts Time of sampling
Before treatment, 30 days after
on the 18th day the beginning
after tumourof the course
inductionof treatment
(48 days after
induction)
Erythrocytes 4,300,000 4,900,000
Leucocytes 20,000 10,000
Thrombocytes 330,000 300,000
The chronic effect of the preparation was tested
on intact rabbits which received the preparation daily in a
dose of 150 mg/kg administered per os. Prior to the experi-
ment as well as on the 10th, 20th, 30th, 60th and 90th days
of study the animals were weighed and blood samples were
taken and analyzed for hemoglobin, erythrocyte count, leucocyte
count, thrombocyte count, residual nitrogen, sugar, total
protein, protein fractions and bilirubin. Throughout the
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entire course of treatment no abnormal behaviour patterns
were observed. The results of the study indicate that on
no count was there any appreciable deviation from normalcy
during the three months of observation. The wglght of the
animals remained stable. On expiration of the three months
the animals were butchered. The autopsy revealed no macrosco-
pic abnormalities. Neither were any abnormalities found in
the histological studies of the tissues of various organs.
The preparation is atoxic in prolonged application.
The preparation was put to clinical trials on 32
patients suffering from oncological diseases of the 4th
degree (cancer of the stomach,Of the rectum, of the mammary
;~` gland, of the uterus and of the lungs; osseous and visceral
sarcomas) with metastases. The preparation was prescribed
per os in doses of from 0.1 to 0.5 g three times a day half
an hour before meals every day for a month, then the treatment
was discontinued for 5 to 10 days, whereupon the course of
treatment was repeated. In spite of the grave initial
-i~ condition, having been put on the preparation, the patients
~ 20 registered general improvement, alleviation or total
; disappearance of pain. The patients often gave up narcotics;
~' their sleep and appetite improved. In some patients, the
tumours diminished and lost in density. The blood picture `
likewise improved.
In accordance with the invention, the preparation
preferably comprises Ringer's solution or physiological
solution as the pharmaceutical solvent for injections. The
;` preparation for injections preferably comprises 5 to 10 percent
: ':
- 30 by weight of the active principle. Applied as tablets, the
preparation comprises 0.1 to 0.5 g of the active principle per
tablet.
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The injection preparation is slowly administered
once a day; in the form of tablets, the preparation is
taken 3 to 4 times a day half an hour before meals.
The preparation is atoxic, has no side effects and is
easily tolerated by the patients, whatever the route of
administration.
The active principle of the preparation, tris-2(hydro-
xyethyl)-ammonium-orthocresoxyacetate, is produced as follows.
Triethanolamine is reacted with o-cresoxyacetic acid
; 10 at a temperature of up to 100C., and the desired product
is subsequently recovered.
The process is effected in an organic solvent miscible
with water.
~ Ethanol, acetone or dioxane are employed as the organic
;~ solvent miscible with water.
In order that the desired product may be of high
~- order of purity, the process is effected in ethanol, and the
desired product is recovered by adding diethyl ether to the
reaction mixture cooled to a temperature of 20 to 30C.
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