New cinnamoyl-cinnamic acid derivative, and its use as pharmaceutical

The present invention relates to a new cinnamoyl-cinnamic acid derivative selected from the group constituted by: PA0 (i) the m-cinnamoyl-cinnamic acid derivatives of formula ##STR1## in which: X.sub.o,X.sub.1,X.sub.2,X.sub.3,X.sub.4, which are identical or different, each represent an atom of hydrogen, a halogen, a lower alkyl group, a lower alkoxy group, the group NRR' (where R and R' identical or different, each represent an atom of hydrogen or a lower alkyl group), the group NO.sub.2, CF.sub.3 or OH; PA1 R.sub.1 represents an atom of hydrogen or a lower alkyl group; PA1 R.sub.2 represents an atom of hydrogen or the methyl group; PA1 Y represents a group OH, OR.sub.3 (where R.sub.3 is a lower alkyl group), NRR' (where R and R' are defined as hereinabove) or the group O(CH.sub.2).sub.n NR.sub.4 R.sub.5 (where n is an integer of value 1 to 5--and preferably 2 or 3)--; and R.sub.4 and R.sub.5, identical or different, each represent an atom of hydrogen, a lower alkyl group and may form with the nitrogen atom to which they are bonded a heterocyclic group of 5 to 7 vertices capable of being substituted and of comprising one or more other heteroatoms such as N and O); PA0 (ii) their geometrical isomers; and PA0 (iii) their salts. It also relates to its method of preparation and its use as pharmaceutical.

The present invention relates as a new industrial product to a chalcone 
which is a cinnamoyl-cinnamic acid derivative. It also relates to its 
method of preparation and to its use as pharmaceutical, particularly as 
growth regulator agent. 
It has been surprizingly found that the compounds of formula I hereinafter, 
which are chalcones belonging to the family of cinnamoyl-cinnamic acid 
derivatives, are useful in therapeutics, particularly as growth regulator 
agents. They allow the restoration of a cellular phenotype of normal cell 
in a transformed cell (for example a cell with anarchic growth) and are 
useful in the treatment of diseases associated with an anarchic 
development of cells and in particular in the treatment of skin disorders 
such as psoriasis, cutaneous keratosis, acne, eczema. 
The new cinnamoyl-cinnamic acid derivative according to the invention is 
characterized in that it is selected from the group constituted by: 
(i) the m-cinnamoyl-cinnamic acid derivatives of formula: 
##STR2## 
in which: X.sub.0, X.sub.1, X.sub.2, X.sub.3, X.sub.4, which are identical 
or different, each represent an atom of hydrogen, a halogen, a lower alkyl 
group a lower alkoxy group, the group NRR' (where R and R', identical or 
different, each represent an atom of hydrogen or a lower alkyl group), the 
group NO.sub.2, CF.sub.3 or OH; 
R.sub.1 represents an atom of hydrogen or a lower alkyl group; 
R.sub.2 represents an atom of hydrogen or the methyl group; 
Y represents an OH group or OR.sub.3 (where R.sub.3 is a lower alkyl 
group), NRR' (where R and R' are defined as hereinabove) or the group 
O(CH.sub.2).sub.n NR.sub.4 R.sub.5 (where n is an integer having a value 
of 1 to 5- and preferably 2 or 3-; and R.sub.4 and R.sub.5, which are 
identical or different, each represent an atom of hydrogen, a lower alkyl 
group and may form with the nitrogen atom to which they are bonded a 
heterocyclic group of 5 to 7 vertices capable of being substituted and of 
comprising one or more other heteroatoms such as N and O); 
(ii) their geometrical isomers; and 
(iii) the salts of the compounds of formula I and of their geometrical 
isomers. 
Lower alkyl group is understood here to mean a branched or straight 
hydrocarbon radical containing 1 to 4 atoms of carbon such as for example 
the methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl or tertiobutyl 
groups. Similarly, lower alkoxy group is understood here to mean a group 
of which the hydrocarbon radical contains 1 to 4 carbon atoms. 
Halogen is understood here to mean an atom of fluorine, an atom of 
chlorine, an atom of bromine or an atom of iodine, the preferred halogen 
atoms being the atom of fluorine and of chlorine. 
Cycle with 5 to 7 vertices which may be substituted and may possibly 
contain one or more other heteroatoms may be constituted, in particular, 
by the pyrrolyl, imidazolyl, pyrazolyl, imidazolidinyl, pyrrolidinyl, 
pyrazolidinyl, piperidyl, piperazinyl, morpholinyl, homopiperidinyl, 
4-methylpiperidyl, 4-methylpiperazinyl, 4-phenylpiperazinyl, 
4-p-chlorophenyl-piperazinyl and 4-.beta.-hydroxyethylpiperazinyl groups. 
The geometrical isomers of the compounds according to the invention may be 
of "cis-cis", "trans-trans", "cis-trans" and "trans-cis" configuration. 
The modi operandi described hereinbelow lead to products which are 
essentially of "trans-trans" configuration, as demonstrated by NMR. 
Salts are understood here to mean (i) the mineral salts obtained from an 
acid of formula I (Y.dbd.OH) with a mineral base (particularly NaOH, KOH, 
NH.sub.3, Ca-OH).sub.2, (ii) the acid addition salts obtained from an acid 
of formula I (Y.dbd.OH) with an organic base and (iii) the acid addition 
salts obtained from a compound of formula I having at least one basic 
radical with a mineral or organic acid. 
The preferred compounds are m-cinnamoyl-cinnamic acid derivatives such as 
Y.dbd.OH, alkoxy with 1 to 4 C atoms, 
.beta.-(4-methylpiperazinyl)-ethyloxy; X.sub.0 .dbd.H, CH.sub.3 ; X.sub.1 
.dbd.H, Cl, OCH.sub.3 ; X.sub.2 .dbd.H, OCH.sub.3 ; X.sub.3 .dbd.H, 
OCH.sub.3 ; X.sub.4 .dbd.H, OCH.sub.3 ; R.sub.1 .dbd.H or CH.sub.3 and 
R.sub.2 .dbd.H. 
The method of preparation recommended according to the invention for 
preparing a compound of formula I is characterised in that an 
acetyl-cinnamic acid derivative of formula: 
##STR3## 
(where X.sub.4, Y, R.sub.1 and R.sub.2 are defined as hereinabove) is 
condensed with a benzaldehyde of formula: 
##STR4## 
(where X.sub.0, X.sub.1, X.sub.2 and X.sub.3 are defined as hereinabove), 
and in that, the geometrical isomers are, if necessary, separated. 
This reaction is advantageously made by reacting IV and V in an aqueous 
solution of an alkali (preferably an aqueous solution of NaOH at 400 g/l), 
or in a lower alcohol with 1 to 4 C atoms saturated by gaseous HCl. This 
technique is perfectly suitable for synthesis of acids I where Y is OH 
when operation is carried out in aqueous solution in the presence of an 
alkali such as sodium hydroxide, from an acid or an ester of formula IV. 
When operation is carried out in a lower alcohol saturated by gaseous HCl, 
the acid of formula IV yields the acid of formula I and, respectively, the 
ester of formula IV yields the ester of formula I; for obtaining an acid, 
the preferred lower alcohol is ethanol and for obtaining an ester, the 
preferred lower alcohol is the one corresponding to the ester group of 
compound IV. 
The acid of formula I (Y.dbd.OH) may, if necessary, be subjected to a 
reaction of esterification or amidification to yield an ester or an amide. 
Similarly, the ester of formula I (Y=alkoxy with 1 to 4 C atoms) may, if 
necessary, be subjected to a reaction of saponification, 
transesterification or transamidification to yield an acid, an ester or an 
amide. 
The separation of the geometrical isomers may be effected either after the 
condensation reaction or after the possible saponifications, 
esterifications and amidifications envisaged hereinabove. If this 
separation is considered necessary, it will preferably be carried out 
after the condensation reaction, the geometrical isomers thus obtained 
then, if necessary, being saponified, esterified or amidified. 
The acetyl-cinnamic acid derivatives of formula IV [which are new 
intermediate products with the exception of the acids (Y.dbd.OH, R.sub.1 
;.dbd.R.sub.2 .dbd.X.sub.4 .dbd.H) and (Y.dbd.OCH.sub.3, X.sub.4 
.dbd.4-OH, and R.sub.1 .dbd.R.sub.2 .dbd.H) described in CA 84, 164 457q 
and respectively CA 89, 108 943m] may be prepared from a derivative of 
2-methyl-2-phenyl-1,3-dioxolan of formula: 
##STR5## 
(where X.sub.4 and R.sub.1 are defined as hereinabove). 
In a first step, a compound of formula II is condensed by means 
(a) either of a triethyl phosphonoalkanoate (and in particular triethyl 
phosphonoacetate or triethyl 2-phosphono-propionate) under the conditions 
of the reaction of HORNER-EMMONS, in the presence of sodium hydride in 
dimethylformamide; 
(b) or of ethyl acetate in ethanol according to CLAISEN in the presence of 
sodium in xylene; to obtain the compound III of formula: 
##STR6## 
In a second step, the deprotection of the dioxolannyl group is effected by 
means of a concentrated acid (preferably 5 N HCl) then, to obtain the 
product IV used as raw material in the process of the invention, a 
saponification is for example carried out to have acid IV, then an 
esterification (or transesterification) to have the ester IV. 
The invention also relates, as new intermediate products, to the compounds 
of formula IV where Y, X.sub.4, R.sub.1 and R.sub.2 are defined as 
hereinabove with the additional condition that at least one of the R.sub.1 
and R.sub.2 is different from H when X.sub.4 is H or OH. 
Non-limiting examples of preparation of cinnamoyl-cinnamic acid derivatives 
of formula I have been given hereinafter.

PREATION I 
Obtaining of 3-[5-(p-chlorocinnamoyl)-2-methoxyphenyl]but-2-ene-oic acid 
(Example 9) 
##STR7## 
(a) Ethyl3-[5-(1,1-ethylenedioxy-ethyl)-2-methoxy-phenyl]-but-2-ene-oate 
At 0.degree. C., 0.24 mole (54 g) of triethyl phosphonoacetate is added, 
drop by drop, to 0.3 mole (7.2 g) of sodium hydride in suspension in 500 
ml of N,N-dimethylformamide. The reaction medium is allowed to return to 
ambient temperature (15.degree.-25.degree. C.) with stirring. After 5 
hours, the emission of hydrogen has finished. The reaction medium is then 
taken to -40.degree. C. and 0.2 mole (47 g) of 
5-(1,1-ethylenedioxy-ethyl)-2-methoxyacetophenone is slowly added. It is 
allowed to return to ambient temperature and stirring is effected for 12 
hours then hydrolysis on ice water. After extraction with ether, washing 
of the ethereal phases up to neutrality, drying and evaporation of the 
solvent, 45 g (yield=73%) of the expected product are collected, in the 
form of oil. 
(b) 3-(5-acetyl-2-methoxy-phenyl)-but-2-ene-oic acid 
45 g (0.147 mole) of the preceding ester are dissolved in 500 ml of ethanol 
and 200 ml of 10% sodium hydroxide. After heating the reaction medium for 
2 hours at reflux, it is left to cool then acidified with 5 N hydrochloric 
acid. The expected product is allowed to precipitate and, after 
filtration, it is washed with water then alcohol. By recrystallisation in 
150 ml of methanol, 15 g of the desired product are obtained 
m.p.=194.degree. C. 
(c) 3-[5-p-chlorocinnamoyl)-2-methoxyphenyl]-but-2-ene-oic acid 
4.7 g (0.02 mole) of the acid previously obtained are dissolved in 30 ml of 
sodium hydroxide at 200 g/l. 2.8 g (0.02 mole) of para-chloro-benzaldehyde 
are added to the reaction medium and it is stirred for 2 hours. After 
acidification of the reaction mixture, the desired product precipitates. 
Filtering is effected, then the precipitate is washed in water. After 
recrystallisation in 300 ml of ethanol, 3 g of pure product are obtained. 
m.p.=215.degree. C. 
PREATION II 
Obtaining of 3-(5-cinnamoyl-2-methoxyphenyl)-but-2-ene-oic acid 
(Example 5) 
##STR8## 
By proceeding as indicated in step (c) of preparation I, from 1 g (4.25 
millimoles) of 3-(5-acetyl-2-methoxy-phenyl)-but-2-ene-oic acid, 10 ml of 
sodium hydroxide at 200 g/l and 0.45 g (4.25 millimoles) of benzaldehyde, 
1.1 g of the desired pure product is obtained. m.p.=190.degree. C. 
PREATION III 
Obtaining of 5-cinnamoyl-2-methoxy-cinnamic acid 
(Example 7) 
##STR9## 
(a) ethyl 3-[5-(1,1-ethylenedioxyethyl)-2-methoxyphenyl]-prop-2-ene-oate 
At 0.degree. C., a mixture of 40 ml of ethyl acetate and 1 cm.sup.3 of 
ethanol is added drop by drop to a suspension of 3.5 g (0.15 mole) of 
sodium in balls in 200 ml of xylene. 22 g (0.1 mole) of 
5-(1,1-ethylenedioxy-ethyl)-2-methoxy benzaldehyde dissolved in 50 ml of 
xylene are then added. The reaction medium is allowed to return to ambient 
temperature and is stirred for 3 hours. 50 ml of acetic acid are then 
added. After total disappearance of the sodium, 40 ml of water are added. 
After extraction with ether, washings with sodium hydroxide then with 
water up to neutrality, the ethereal phases are evaporated to obtain 24 g 
(oil) of the expected product. 
(b) ethyl 3-(5-acetyl-2-methoxyphenyl)prop-2-ene oate 
9.6 g of the ester obtained in step (a) are dissolved in 150 ml of ether 
and 20 ml of 5 N hydrochloric acid are added. Stirring is carried out for 
12 hours. After extraction by the (1:1) v/v ether-CH.sub.2 Cl.sub.2 
mixture, then washings with water, 7.5 g of the expected product are 
obtained after evaporation of the organic phases. m.p.=90.degree. C. 
(c) 3-(5-acetyl-2-methoxyphenyl)-prop-2-ene-oic acid 
24 g (0.08 mole) of the ester obtained in step (b) are dissolved in 250 ml 
of methanol and 50 ml of sodium hydroxide at 400 g/l. After the reaction 
mixture is heated for 2 hours under reflux, it is left to cool then 
acidified with 5 N hydrochloric acid. The expected product is left to 
precipitate and, after filtration, is washed with water then with ethanol 
and finally with isopropyl ether. 13.6 g of the desired product are 
obtained. m.p.=203.degree. C. 
(d) 3-(5-cinnamoyl-2-methoxyphenyl)-prop-2-ene oic acid 
Using 10 g (0.045 mole) of the acid obtained in step (c), 80 ml of 20% 
sodium hydroxide and 5 g (0.045 mole) of benzaldehyde and according to a 
modus operandi similar to the one described in step (c) of preparation I, 
11.5 g of the desired pure product are obtained, m.p.=204.degree. C. 
PREATION IV 
Obtaining of ethyl 3-[m-(p-chlorocinnamoyl]-prop-2-ene-oate 
(Example 3) 
##STR10## 
other nomenclature: ethyl m-(p-chlorocinnamoyl)-cinnamate 
12 g (0.055 mole) of the ethyl ester of 3-acetyl-cinnamic acid are 
dissolved in 120 ml of ethanol. 8 g (0.057 mole) of 
para-chlorobenzaldehyde are added then 35 ml of anhydrous ethanol 
saturated with gaseous hydrochloric acid. The reaction medium is stirred 
for 10 hours. 
By cooling the reaction medium to 0.degree. C., the desired product 
precipitates. It is filtered then washed with hexane. 13.5 g (yield=72%) 
of the expected pure product are thus obtained. m.p. 138.degree. C. 
A certain number of compounds according to the invention have been included 
in Table I hereinafter, in non-limiting manner. 
The products according to the invention are used in therapeutics as agents 
promoting the restoration of a cellular phenotype of normal cell in a 
transformed strain. They may be used in particular for the topical or 
systemic treatment of skin disorders such as for example cutaneous 
keratoses, acne, psoriasis, eczemas, warts or any other skin disorder 
involving an alteration of the corneal tissue and for the treatment of 
inflammatory or degenerative alterations of the mucous membranes, 
cartilages, muscles or tendons, such as for example arthrosis, infectious 
rheumatism. 
It will be recalled that, when normal cells (healthy cells) are cultivated 
on a given nutrient medium in a given space, it is observed that, after a 
certain lapse of time, the number of cells present is constant 
(confluence) and that, when transformed cells (abnormal cells of anarchic 
development) are cultivated under the same conditions, a constant number 
of cells is not obtained, as cellular proliferation does not cease 
(absence of confluence). 
The pharmacological results obtained on a test for restoring the inhibition 
of growth with confluence of a culture of transformed cells are summarised 
hereinafter. 
The modus operandi used is the one described by L. D. DION et al in J. 
Natl. Cancer inst. 58 (No. 3), pages 795-801 (1977) and summed up below. 
20 000 murine cells are seeded on a surface of 0.2 cm.sup.2 on a culture 
medium "Minimum Essential Medium" with Eagle salts and 10% of foetal calf 
serum. After 4 hours, the cells have adhered to the support. The culture 
medium is then replaced by the same medium containing the product to be 
tested dissolved in DM50. 
At regular intervals over a period of two weeks, the cells are counted 
after trypsination. The growth curve obtained is compared with the one 
obtained in the absence of substance to be tested. 
The effect of the drug is assessed by the minimum concentration necessary 
for inhibiting any growth from the moment when confluence is obtained. 
The results obtained are given in Table II hereinafter which also shows the 
results relative to toxicity (LD-0 or LD-50) in the mouse by the 
intraperitoneal route. 
An additional test was made with the product of Example 9, according to the 
test of inhibition of TPA-induced ornithine-decarboxylase activity (TPA 
being 12-O-tetradecanoylphorbol-13-acetate), described by A. K. VERMA et 
al in Cancer Research 38, 793-801 (1978). 
The product in solution in 100 .mu.l of acetone is tested by topical 
application on mice, an hour before topical application of 34 nmoles of 
TPA. The activity of the ornithine-decarboxylase is measured 4.5 hours 
after treatment with TPA. Under these conditions, an inhibition of 48% of 
the activity of the ornithine-decarboxylase is obtained by topical 
administration of 34 nmoles of the product of Example 9. 
According to the invention, a therapeutical composition is recommended, 
characterised in that it contains, in association with a physiologically 
acceptable excipient, at least one compound of formula I, one of its 
geometrical isomers or one of their pharmaceutically acceptable salts. 
The daily dosage which is recommended for the compounds according to the 
invention is from 0.1 to 50 mg per kg of body weight (preferably from 0.1 
to 5 mg/kg) in the form of capsules or tablets, for oral administration. 
TABLE I 
__________________________________________________________________________ 
##STR11## 
Melting point 
Example 
X.sub.o 
X.sub.1 
X.sub.2 
X.sub.3 
X.sub.4 
R.sub.1 
R.sub.2 
Y (0.degree. C.) 
__________________________________________________________________________ 
1 H 4-Cl H H H H H OH 250 
2 H 3-OCH.sub.3 
4-OCH.sub.3 
5-OCH.sub.3 
H H H OC.sub.2 H.sub.5 
133 
3 H 4-Cl H H H H H OC.sub.2 H.sub.5 
138 
4 H H H H H H H OH 188 
5 H H H H 2-OCH.sub.3 
CH.sub.3 
H OH 190 
6 H H H H 2-OCH.sub.3 
H H 
##STR12## 180 
7 H H H H 2-OCH.sub.3 
H H OH 209 
8 H 4-OCH.sub.3 
H H 2-OCH.sub.3 
H H OH 230 
9 H 4Cl H H 2-OCH.sub.3 
CH.sub.3 
H OH 215 
10 H 4-Cl H H 2-OCH.sub.3 
H H OH 242 
11 H 4-Cl H H 2-OCH.sub.3 
CH.sub.3 
H NH.sub.2 199 
12 H 4-CH.sub.3 
H H 2-OCH.sub.3 
CH.sub.3 
H OH 210 
13 H 4-Cl H H 2-OCH.sub.3 
H CH.sub.3 
OH 222 
14 H 3-OCH.sub.3 
4-OCH.sub.3 
5-OCH.sub.3 
2-OCH.sub.3 
CH.sub.3 
H OC.sub.2 H.sub.5 
120 
15 H 2-CH.sub.3 
H H 2-OCH.sub.3 
CH.sub.3 
H OH 202 
16 H 2-Cl H H 2-OCH.sub.3 
CH.sub.3 
H OH 232 
17 H 3-Cl 4-Cl H 2-OCH.sub.3 
CH.sub.3 
H OH 212 
18 2-CH.sub.3 
3-CH.sub.3 
4-OCH.sub.3 
6-CH.sub.3 
2-OCH.sub.3 
CH.sub.3 
H OH 117 
19 H 4-Cl H H 2-OCH.sub.3 
CH.sub.3 
H OC.sub.2 H.sub.5 
129 
20 H 4-Cl H H 2-OCH.sub.3 
CH.sub.3 
H O(CH.sub.2).sub.2 NEt.sub.2 
120 
21 H 4-Cl H H 2-OCH.sub.3 
CH.sub.3 
H 
##STR13## 140 
22 H 4-Cl H H H CH.sub.3 
H OH 189 
__________________________________________________________________________ 
The proof of the "transtrans" configuration of the product of Example 18 
has been brought by NMR study which gives: 
- double bond CC of the chalcone group J.sub.H--H = 16 Hz 
- double bond CC of the cinnamic group J.sub.H--H = 1.1 Hz 
TABLE II 
______________________________________ 
Concentration of inhib- 
ition of growth with 
i.p. toxicity 
confluence 
Example mice (mg/kg) 
in .mu.g/ml 
______________________________________ 
1 DL-0 .gtoreq.1600 
5 to 10 
2 DL-0 .gtoreq.1600 
10 
3 DL-0 .gtoreq.1600 
-- 
4 DL-50 = 450 &gt;10 
5 DL-50 = 550 5 to 10 
6 DL-50 = 300 -- 
7 DL-50 = 1000 
-- 
8 DL-0 .gtoreq.800 
&gt;10 
9 DL-0 .gtoreq.1600 
5 
______________________________________ 
.+-. = Reference product: retinoic acid: 5 .mu.g/ml