U.S. patent application number 09/801664 was filed with the patent office on 2001-08-30 for use of a rar-gamma-specific agonist ligand for increasing the rate of apoptosis.
Invention is credited to Fesus, Laszlo, Reichert, Uwe, Szondy, Zsuzsa.
Application Number | 20010018456 09/801664 |
Document ID | / |
Family ID | 9483613 |
Filed Date | 2001-08-30 |
United States Patent
Application |
20010018456 |
Kind Code |
A1 |
Fesus, Laszlo ; et
al. |
August 30, 2001 |
Use of a RAR-gamma-specific agonist ligand for increasing the rate
of apoptosis
Abstract
A method for increasing the rate of apoptosis in at least one
cell population in which apoptosis is induced by activating
receptors of the RAR-.gamma. type, the method comprising exposing
the cell population to an effective amount of at least one agonist
ligand which is specific for receptors of the RAR-.gamma. type to
increase the rate of apoptosis.
Inventors: |
Fesus, Laszlo; (Debrecen,
HU) ; Szondy, Zsuzsa; (Debrecen, HU) ;
Reichert, Uwe; (Pont-du-Loup, FR) |
Correspondence
Address: |
Norman H. Stepno
BURNS, DOANE, SWECKER & MATHIS, L.L.P.
P.O. Box 1404
Alexandria
VA
22313-1404
US
|
Family ID: |
9483613 |
Appl. No.: |
09/801664 |
Filed: |
March 9, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09801664 |
Mar 9, 2001 |
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09051407 |
Jul 15, 1998 |
|
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09051407 |
Jul 15, 1998 |
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PCT/FR96/01568 |
Oct 8, 1996 |
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Current U.S.
Class: |
514/569 |
Current CPC
Class: |
Y10S 514/826 20130101;
Y10S 514/844 20130101; A61P 43/00 20180101; Y10S 514/866 20130101;
Y10S 514/903 20130101; A61P 37/00 20180101; A61P 37/08 20180101;
Y10S 514/825 20130101; A61P 17/06 20180101; Y10S 514/863 20130101;
A61P 3/08 20180101; A61P 1/16 20180101; A61P 3/10 20180101; A61P
31/12 20180101; A61P 17/00 20180101; A61K 31/192 20130101; A61P
29/00 20180101; A61P 35/00 20180101 |
Class at
Publication: |
514/569 |
International
Class: |
A61K 031/19; A01N
037/10 |
Foreign Application Data
Date |
Code |
Application Number |
Oct 9, 1995 |
FR |
95/12179 |
Claims
What is claimed is:
1. A method for increasing the rate of apoptosis in at least one
cell population in which apoptosis is induced by activating
receptors of the RAR-.gamma. type, said method comprising exposing
said cell population to an effective amount of at least one agonist
ligand which is specific for receptors of the RAR-.gamma. type to
increase the rate of apoptosis, with the proviso that said agonist
ligand is not 6-3-(1-adamantyl)-4-hydroxyph- enyl-2-naphthanoic
acid, said method being used to treat a precancerous condition
linked to an inadequate rate of apoptosis.
2. The method according to claim 1, wherein said precancerous
condition is actinic keratosis.
3. A method for increasing the rate of apoptosis in at least one
cell population in which apoptosis is induced by activating
receptors of the RAR-.gamma. type, said method comprising exposing
said cell population to an effective amount of at least one agonist
ligand which is specific for receptors of the RAR-.gamma. type to
increase the rate of apoptosis, wherein said agonist ligand which
is specific for receptors of the RAR-.gamma. type exhibits a ratio
R which is greater than or equal to 50, wherein R is the
dissociation constant of the ligand for receptors of the
RAR-.alpha. type relative to receptors of the RAR-.gamma. type,
with the proviso that said agonist ligand is not
6-3-(1-adamantyl)-4-hydroxyphenyl- -2-naphthanoic acid, said method
being used to treat a precancerous condition linked to an
inadequate rate of apoptosis.
4. The method according to claim 3, wherein said precancerous
condition is actinic keratosis.
5. A method for increasing the rate of apoptosis in at least one
cell population in which apoptosis is induced by activating
receptors of the RAR-.gamma. type, said method comprising exposing
said cell population to an effective amount of at least one agonist
ligand which is specific for receptors of the RAR-.gamma. type to
increase the rate of apoptosis, said agonist ligand being selected
from the group consisting of
(E)-4-(1-hydroxyl-1-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-2-
-propenyl)benzoic acid,
4-[(E)-2-(3-(1-adamantyl)-4-hydroxyphenyl)-1-prope- nyl]benzoic
acid, 5',5',8',8'-tetramethyl-5',6',7',8'-tetrahydro-[2,2']bin-
aphthalenyl-6-carboxylic acid,
2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydrona-
phthalen-2-yl)benzo[b]thiophene-6-carboxylic acid,
4-(5,5,8,8-tetramethyl-- 5,6,7,8-tetrahydronaphtho[
2,3-b]thiophen-2-yl)benzoic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalene-2-carbonyl)naphthale-
ne-2-carboxylic acid,
3,7-dimethyl-7-(1,2,3,4-tetrahydro-1,4a,9b-trimethyl-
-1,4-methano-dibenzofuran-8-yl)-2,4,6-heptatrienoic acid,
6-(1,2,3,4-tetrahydro-1,4a,9b-trimethyl-1,4-methano-dibenzofuran-8-yl)nap-
hthalene-2-carboxylic acid, 6-[
hydroxyimino-(5,5,8,8-tetramethyl-5,6,7,8--
tetrahydro-naphthalen-2-yl)methyl]naphthalene-2-carboxylic acid,
4-[(6-hydroxy-7-(1-adamantyl)-2-naphthyl]benzoic acid,
5-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroanthracen-2-yl)thiophene-2-carbo-
xylic acid,
(-)-6-[hydroxy-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthal-
en-2-yl)methyl] naphthalene-2-carboxylic acid,
6-(3-adamantan-1-yl-4-prop--
2-ynyloxyphenyl)naphthalene-2-carboxylic acid,
4-[(2-oxo-2-(5,5,8,8-tetram-
ethyl-5,6,7,8-tetrahydronaphthalen-2-yl)ethoxy]benzoic acid,
4-[2-oxo-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)acetyla-
mino]benzoic acid, 4-[
2-fluoro-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro--
naphthalen-2-yl)acetylamino]benzoic acid,
6-[3-(1-adamantyl-4-(2-hydroxypr- opyl)phenyl]-2-naphthoic acid,
5-[3-oxo-3-(5,5,8,8-tetramethyl-5,6,7,8-tet-
rahydro-naphthalen-2-yl)propenyl]thiophene-2-carboxylic acid,
6-[3-(1-adamantyl-4-(2,3-dihydroxypropyl)phenyl]-2-naphthoic acid,
4-[3-hydroxy-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-1-propyn-
yl]benzoic acid,
4-[3-oxo-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphtha-
len-2-yl)prop-1-ynyl]benzoic acid, 4[
(3-(1-methylcyclohexyl)-4-hydroxyphe- nyl)ethenyl]-benzoic acid,
4-[(E)2-(3-(1-adamantyl)-4-hydroxyphenyl)etheny- l] benzoic acid,
4-[3-(1-adamantyl)-4-hydroxyphenylethynyl)benzoic acid,
5-[3-(1-adamantyl)-4-hydroxyphenylethynyl]-2-thiophenecarboxylic
acid,
5-[3-(1-adamantyl)-4-methoxyphenylethynyl]-2-thiophenecarboxylic
acid, 4-[2-(3-tert-butyl-4-methoxyphenyl)propenyl]benzoic acid,
4-{2-[4-methoxy-3-(1-methylcyclohexyl)phenyl]-propenyl} benzoic
acid,
6-[3-(1-adamantyl)-4-(3-methoxy-2-hydroxypropyl)phenyl]-2-naphthoic
acid, 2-hydroxy-4-[
3-hydroxy-3-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napht-
hyl)-1-propynyl]benzoic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronap-
hthalen-2-yloxy)naphthalene-2-carboxylic acid,
6-(5,5,8,8-tetramethyl-5,6,-
7,8-tetrahydronaphthalen-2-ylsulphanyl)naphthalene-2-carboxylic
acid,
4-[2-propoxyimino-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-naphthalen-2--
yl)acetylamino]benzoic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaph-
thalen-2-ylamino)naphthalene-2-carboxylic acid,
1-methyl-4-(5,5,8,8-tetram-
ethyl-5,6,7,8-tetrahydroanthracen-2-yl)-1H-pyrrole-2-carboxylic
acid,
2-methoxy-4-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroanthracen-2-yl)benzoic
acid,
4-[2-nonyloxyimino-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphtha-
len-2-yl)acetylamino]benzoic acid,
(-)-2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-t-
etramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)prop-1-ynyl] benzoic
acid,
(+)-2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronapht-
halen-2-yl)prop-1-ynyl]benzoic acid,
2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tet-
ramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)but-1-ynyl]benzoic acid,
6-(3-bromo-5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yloxy)-naph-
thalene-2-carboxylic acid,
3-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-na-
phthyl)-2H-1-benzopyran]-7-carboxylic acid,
4-[3-(3,5-di-tert-butyl-4-hydr- oxyphenyl)prop-1-ynyl]benzoic acid,
4-[3-(5,5,8,8-tetramethyl-5,6,7,8-tetr-
ahydronaphthalen-2-yl)prop-1-ynyl] benzoic acid,
4-[3-(5,6,7,8-tetrahydro--
5,5,8,8-tetramethyl-2-naphthyl)-1-propynyl]salicylic acid, 4-[
{3-(1-adamantyl)-4-(2-hydroxyethyl)phenyl}ethynyl]-benzoic acid and
4-[{3-(1-adamantyl)-4-(3-hydroxypropyl)phenyl} ethynyl]benzoic
acid, said method being used to treat a precancerous condition
linked to an inadequate rate of apoptosis.
6. The method according to claim 5, wherein said precancerous
condition is actinic keratosis.
7. A method for increasing the rate of apoptosis in at least one
cell population in which apoptosis is induced by activating
receptors of the RAR-.gamma.-type, said method comprising exposing
said cell population to an effective amount of at least one agonist
ligand which is specific for receptors of the RAR-.gamma. type to
increase the rate of apoptosis, said agonist ligand being selected
from the group consisting of
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalene-2-carbonyl)-naphthal-
ene-2-carboxylic acid,
6-(1,2,3,4-tetrahydro-1,4a,9b-trimethyl-1,4-methano-
dibenzofuran-8-yl)-naphthalene-2-carboxylic acid,
6-[hydroxyimino-(5,5,8,8-
-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)methyl]
naphthalene-2-carboxylic acid,
5-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroa-
nthracen-2-yl)-thiophene-2-carboxylic acid,
(-)-6-[hydroxy-(5,5,8,8-tetram-
ethyl-5,6,7,8-tetrahydronaphthalen-2-yl)-methyl]
naphthalene-2-carboxylic acid,
6-[3-(1-adamantyl-4-(2-hydroxypropyl)phenyl]-2-naphthoic acid,
6-[3-(1-adamantyl-4-(2,3-dihydroxypropyl)phenyl]-2-naphthoic acid,
4-[3-(1-adamantyl)-4-hydroxyphenylethynyl] benzoic acid,
5-[3-(1-adamantyl)-4-hydroxyphenylethynyl]-2-thiophenecarboxylic
acid, 5-[
3-(1-adamantyl)-4-methoxyphenylethynyl]-2-thiophenecarboxylic acid,
6-[3-(1-adamantyl)-4-(3-methoxy-2-hydroxypropyl)phenyl]-2-naphthoic
acid,
1-methyl-4-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-anthracen-2-yl)-1H-pyr-
role-2-carboxylic acid,
(-)-2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl--
5,6,7,8-tetrahydronaphthalen-2-yl)prop-1-ynyl]benzoic acid and
2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthale-
n-2-yl)but-1-ynyl)benzoic acid, said method being used to treat a
precancerous condition linked to an inadequate rate of
apoptosis.
8. The method according to claim 7, wherein said precancerous
condition is actinic keratosis.
9. The method according to claim 1, wherein said agonist ligand is
used in combination with a pharmaceutically acceptable carrier.
10. The method according to claim 3, wherein said agonist ligand is
used in combination with a pharmaceutically acceptable carrier.
11. The method according to claim 5, wherein said agonist ligand is
used in combination with a pharmaceutically acceptable carrier.
12. The method according to claim 7, wherein said agonist ligand is
used in combination with a pharmaceutically acceptable carrier.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of copending U.S.
Application Ser. No. 09/051,407, filed Jul. 15, 1998, incorporated
by reference herein in its entirety and relied upon, which is the
U.S. national stage of International Application No.
PCT/FR96/01568, filed Oct. 8, 1996.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to the use of specific
retinoids in the preparation of a pharmaceutical composition which
is intended to increase the rate of apoptosis. These retinoids can
also be used in cosmetic compositions which are intended, in
particular, to prevent and/or combat photoinduced or chronological
ageing of the skin.
[0003] Two types of mechanism are involved in the death of cells.
The first, which is the classical type, is termed necrosis.
Morphologically, necrosis is characterized by swelling of the
mitochondria and the cytoplasm and by nuclear distortion, followed
by destruction of the cell and its autolysis, with the latter being
accompanied by an inflammation phenomenon. Necrosis occurs in a
passive and incidental manner. Tissue necrosis is generally due to
the cells being subjected to a physical trauma, or due to a
chemical poison, for example.
[0004] The other form of cell death is termed apoptosis [Kerr, J.
F. R. and Wyllie, A. H., Br. J. Cancer, 265, 239 (1972)]; however,
contrary to necrosis, apoptosis does not result in any inflammation
phenomenon. Apoptosis has been reported to be able to take place
under various physiological conditions. It is a highly selective
form of cell suicide which is characterized by readily observable
morphological and biochemical phenomena. Thus, condensation of the
chromatin, which is or is not associated with an endonuclease
activity, formation of apoptotic bodies and fragmentation of the
deoxyribonucleic acid (DNA), by activation of endonucleases, into
180-200 base pair DNA fragments (these fragments can be observed by
means of agarose gel electrophoresis) are, in particular,
observed.
[0005] Apoptosis can be regarded as being a programmed cell death
which is involved in tissue development, differentiation and
renewal. It is also thought that the differentiation, growth and
maturation of cells are closely linked to apoptosis and that the
substances which are able to play a role in the differentiation,
growth and maturation of cells are also linked to the phenomenon of
apoptosis.
[0006] In the medical field, some pathological situations exhibit a
modified, if not deregulated, apoptosis mechanism. Thus, it has
been reported that deliberate modulation of apoptosis, by inducing
it or suppressing it, can make it possible to treat a large number
of diseases, more specifically diseases linked to cell
hyperproliferation, as in the case of cancer, autoimmune diseases
and allergies, or, on the other hand, diseases which are linked to
cell disappearance, as in the case of the human immunodeficiency
virus (HIV) immunodeficiency syndrome, neurodegenerative diseases
(Alzheimer's disease) or excessive damage which is induced during
myocardial infarction.
[0007] Specifically, it has been noted in oncology that a large
number of antineoplastic drugs, such as dexamethasone,
cyclophosphamide and cisplatin, are able to induce apoptosis.
[0008] In the cosmetic field, the signs of cutaneous ageing
essentially result from dysfunction of the principal biological
mechanisms of the skin which, in particular, bring the mechanism of
apoptosis into play. It is possible, therefore, to imagine that any
product which induces the mechanism of apoptosis is a product which
is suitable for preventing and/or combatting the appearance of
ageing and the existing signs of ageing such as large and small
wrinkles.
[0009] In the field of retinoids, it is known that all-trans
retinoic acid is a powerful modulator (i.e. an inhibitor or, on the
other hand, a stimulator, depending on the nature of the cells
which are treated) of the differentiation and proliferation of many
normal or transformed cell types. For example, it inhibits the
differentiation of epithelial cells such as the keratinocytes of
the epidermis. It also inhibits the proliferation of many
transformed cells such as melanoma cells. These effects on
proliferation and differentiation can affect one and the same type
of cell simultaneously, as is the case, for example, for HL-60
human promyelocytic cells; thus, it is known that proliferation of
these cells is inhibited by all-trans retinoic acid and that, at
the same time, their differentiation into granulocytes and their
apoptosis are induced.
[0010] It is known, in a general manner, that all-trans retinoic
acid acts on the differentiation and proliferation of cells by
interacting with nucleoreceptors which are termed RARs (retinoic
acid receptors) and which are present in the cell nucleus. To date,
three subtypes of RAR receptors, termed RAR-.alpha., RAR-.beta. and
RAR-.gamma., respectively, have been identified. After having bound
the ligand (i.e. all-trans retinoic acid), these receptors interact
with specific response elements (RARE) in the promoter region of
genes which are regulated by retinoic acid. In order to bind to the
response elements, the RARs heterodimerize with another type of
receptor known as RXR receptors. The natural ligand of the RXRs is
9-cis-retinoic acid. The RXRs are regarded as being master
regulatory proteins because they interact with other members of the
steroid/thyroid receptor superfamily, such as the receptor for
vitamin D3 (VDR), the receptor for triiodothyroxine (TR) and the
PPARs (peroxisome proliferator activated receptors), to form
heterodimers, as they do with the RARs. Furthermore, the RXRs are
able to interact with specific response elements (RXRE) in the form
of homodimers. These complex interactions, and the existence of
numerous RAR and RXR receptors which are expressed differently
depending on the tissue and the cell type, explain the pleiotropic
effects of retinoids in virtually all cells.
[0011] Large numbers of synthetic structural analogues of all-trans
retinoic acid or of 9-cis-retinoic acid, commonly termed
"retinoids", have so far been described in the literature. Some of
these molecules are able to bind to, and specifically activate, the
RARs or, on the other hand, the RXRs. Furthermore, some analogues
are able to bind to, and activate, a particular subtype (.alpha.,
.beta. or .gamma.) of RAR receptor. Finally, other analogues do not
exhibit any particular selective activity with regard to these
different receptors. In this respect, and by way of example,
9-cis-retinoic acid activates both the RARs and the RXRs without
any noteworthy selectivity for either of these receptors
(nonspecific agonist ligand), whereas all-trans retinoic acid
selectively activates the RARs (RAR-specific agonist ligand)
without regard to subtype. In a general manner, and qualitatively,
a given substance (or ligand) is said to be specific for a given
receptor family (or with regard to a particular receptor of this
family) when the said substance exhibits an affinity for all the
receptors of this family (or, respectively, for the particular
receptor of this family) which is stronger than that which it
otherwise exhibits for all the receptors of any other family (or,
respectively, for all the other receptors, of this same family or
not).
[0012] It has been reported that 9-cis-retinoic acid and all-trans
retinoic acid are modulators of apoptosis (activator or inhibitor
of apoptosis depending, in particular, on the cell type) and that
9-cis-retinoic acid is the more active of these two modulators,
with it being possible to explain this observation by the fact that
9-cis-retinoic acid activates both the RARs and the RXRs, contrary
to all-trans retinoic acid, which only activates the RARs.
[0013] In view of that which has been previously stated, it appears
to be of interest to find novel modulators of apoptosis.
BRIEF SUMMARY OF THE INVENTION
[0014] In this regard, the Applicant has just discovered that
agonist ligands which are specific for receptors of the RAR-.gamma.
type are excellent inducers of apoptosis in a variety of cell
types, more specifically in thymocytes.
[0015] Thus, the present invention relates to the use of at least
one agonist ligand which is specific for receptors of the
RAR-.gamma. type in the preparation of a pharmaceutical composition
which is intended to increase the rate of apoptosis in at least one
cell population in which apoptosis can be induced by activating
receptors of the RAR-.gamma. type.
[0016] The invention also relates to the use, in a cosmetic
composition, of at least one agonist ligand which is specific for
receptors of the RAR-.gamma. type as an inducer of apoptosis in at
least one cell population of the skin in which apoptosis can be
induced by activating receptors of the RAR-.gamma. type.
[0017] Thus, this composition can make it possible to prevent
and/or combat photoinduced or chronological ageing of the skin, in
particular by eliminating, by means of apoptosis, cells of the skin
which exhibit a deficiency in their repair function and which
accumulate over time.
[0018] The invention therefore relates, finally, to a cosmetic
process for preventing and/or combatting photoinduced or
chronological ageing of the skin, characterized in that an
apoptosis-inducing cosmetic composition, such as previously
described, is applied to the skin.
DETAILED DESCRIPTION OF THE INVENTION
[0019] The pharmaceutical or cosmetic composition according to the
invention comprises a physiologically acceptable medium.
[0020] Agonist ligand which is specific for receptors of the
RAR-.gamma. type is understood, according to the invention, as
meaning any agonist ligand which exhibits a ratio R of the
dissociation constant of this ligand for receptors of the
RAR-.gamma. type over the dissociation constant of this ligand for
receptors of the RAR-.gamma. type which is greater than or equal to
10, and which induces differentiation of F9 cells.
[0021] Thus, it is known that all-trans retinoic acid and some of
its analogues are able to induce the differentiation of mouse
embryonic teratocarcinoma cells (F9 cells); they are therefore
regarded as being agonists for the RAR receptors. The secretion of
the plasminogen activator which accompanies this differentiation is
an index of the biological response of the F9 cells to the
retinoids (Skin pharmacol. 1990; 3: pp. 256-267).
[0022] The dissociation constants are determined by means of tests
which are standard for the skilled person. These tests are
described, in particular, in the following references: (1)
"Selective Synthetic Ligands for Nuclear Retinoic Acid Receptor
Subtypes" in RETINOIDS, Progress in Research and Clinical
Applications, Chapter 19 (pp 261-267), Marcel Dekker Inc., edited
by Maria A. Livrea and Lester Packer; (2) "Synthetic Retinoids:
Receptor Selectivity and Biological Activity" in Pharmacol Skin,
Basel, Karger, 1993, Volume 5, pp 117-127; (3) "Selective Synthetic
Ligands for Human Nuclear Retinoic Acid Receptors" in Skin
Pharmacology, 1992, Vol. 5, pp 57-65; (4) "Identification of
Synthetic Retinoids with Selectivity for Human Nuclear Retinoic
Acid Receptor-.gamma." in Biochemical and Biophysical Research
Communications, Vol. 186, No. 2, July 1992, pp 977-983; (5)
"Selective High Affinity RAR-.alpha. or RAR-.beta. Retinoic Acid
Receptor Ligands" in Mol. Pharmacol., Vol. 40, pp 556-562.
[0023] Other characteristics, aspects, aims and advantages of the
invention will become even clearer from reading the description
which follows as well as the various specific examples, which are
intended as an illustration and in no way as being limiting.
[0024] Agonist ligands which are specific for receptors of the
RAR-.gamma. type, and which may be mentioned, are
6-3-(1-adamantyl)-4-hydroxyphenyl)-- 2-naphthanoic acid,
(E)-4-(1-hydroxy-1-(5,6,7,8-tetrahydro-5,5,8,8-tetrame-
thyl-2-naphthyl)-2-propenyl)benzoic acid,
4-[(E)-2-(3-(1-adamantyl)-4-hydr- oxyphenyl)-1-propenyl]benzoic
acid, 5',5',8',8'-tetramethyl-5',6',7',8'-te- trahydro-[
2,2']binaphthalenyl-6-carboxylic acid, 2-(5,5,8,8-tetramethyl-5-
,6,7,8-tetrahydronaphthalen-2-yl)benzo[ b]thiophene-6-carboxylic
acid,
4-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphtho[2,3-b]thiophen-2-yl)benz-
oic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalene-2-carbonyl)-
naphthalene-2-carboxylic acid,
3,7-dimethyl-7-(1,2,3,4-tetrahydro-1,4a,9b--
trimethyl-1,4-methanodibenzofuran-8-yl)-2,4,6-heptatrienoic acid,
6-(1,2,3,4-tetrahydro-1,4a,9b-trimethyl-1,4-methanodibenzofuran-8-yl)naph-
thalene-2-carboxylic acid,
6-[hydroxyimino-(5,5,8,8-tetramethyl-5,6,7,8-te-
trahydronaphthalen-2-yl)methyl]naphthalene-2-carboxylic acid,
4-[(6-hydroxy-7-(1-adamantyl)-2-naphthyl]benzoic acid,
5-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroanthracen-2-yl)thiophene-2-carbo-
xylic acid,
(-)-6-[hydroxy-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthal-
en-2-yl)methyl]naphthalene-2-carboxylic acid,
6-(3-adamantan-1-yl-4-prop-2-
-ynyloxyphenyl)naphthalene-2-carboxylic acid,
4-[(2-oxo-2-(5,5,8,8-tetrame-
thyl-5,6,7,8-tetrahydronaphthalen-2-yl)ethoxy]benzoic acid,
4-[2-oxo-2-(5,5,8,8-tetramethyl-5,6,7,8-tetra-hydronaphthalen-2-yl)acetyl-
amino]benzoic acid,
4-[2-fluoro-2-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydron-
aphthalen-2-yl)acetylamino]benzoic acid,
6-[3-(1-adamantyl-4-(2-hydroxypro- pyl)phenyl]-2-naphthoic acid,
5-[3-oxo-3-(5,5,8,8-tetramethyl-5,6,7,8-tetr-
ahydronaphthalen-2-yl)propenyl]thiophene-2-carboxylic acid,
6-[3-(1-adamantyl-4-(2,3-di-hydroxypropyl)phenyl]-2-naphthoic acid,
4-[3-hydroxy-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-1-propyn-
yl]benzoic acid,
4-[3-oxo-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphtha-
len-2-yl)prop-1-ynyl]benzoic acid,
4-[(3-(1-methylcyclohexyl)-4-hydroxyphe- nyl)ethenyl]benzoic acid,
4-[(E)2-(3-(1-adamantyl)-4-hydroxyphenyl)ethenyl- ]benzoic acid,
4-[3-(1-adamantyl)-4-hydroxyphenylethynyl)benzoic acid,
5-[3-(1-adamantyl)-4-hydroxyphenylethynyl]-2-thiophenecarboxylic
acid,
5-[3-(1-adamantyl)-4-methoxyphenylethynyl]-2-thiophenecarboxylic
acid, 4-[2-(3-tert-butyl-4-methoxyphenyl)propenyl]benzoic acid,
4-{2-[4-methoxy-3-(1-methylcyclohexyl)phenyl] propenyl}benzoic
acid,
6-[3-(1-adamantyl)-4-(3-methoxy-2-hydroxypropyl)-phenyl]-2-naphthoic
acid,
2-hydroxy-4-[3-hydroxy-3-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2--
naphthyl)-1-propynyl]benzoic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahyd-
ronaphthalen-2-yloxy)naphthalene-2-carboxylic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-ylsulphanyl)naphtha-
lene-2-carboxylic acid,
4-[2-propoxyimino-2-(5,5,8,8-tetramethyl-5,6,7,8-t-
etrahydronaphthalen-2-yl)-acetylamino] benzoic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-ylamino)naphthalene-
-2-carboxylic acid,
1-methyl-4-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroanth-
racen-2-yl)-1H-pyrrole-2-carboxylic acid,
2-methoxy-4-(5,5,8,8-tetramethyl-
-5,6,7,8-tetrahydroanthracen-2-yl)benzoic acid,
4-[2-nonyloxyimino-2-(5,5,-
8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)acetylamino]benzoic
acid,
(-)-2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydr-
onaphthalen-2-yl)prop-1-ynyl]benzoic acid,
(+)-2-hydroxy-4-[3-hydroxy-3-(5-
,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)prop-1-ynyl]benzoic
acid,
2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronap-
hthalen-2-yl)but-1-ynyl]benzoic acid,
6-(3-bromo-5,5,8,8-tetramethyl-5,6,7-
,8-tetrahydronaphthalen-2-yloxy)naphthalene-2-carboxylic acid,
3-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-2H-1-benzopyran]-7-
-carboxylic acid,
4-[3-(3,5-di-tert-butyl-4-hydroxyphenyl)prop-1-ynyl]benz- oic acid,
4-[3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)prop-
-1-ynyl]benzoic acid,
4-[3-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napht-
hyl)-1-propynyl]salicylic acid,
4-[{3-(1-adamantyl)-4-(2-hydroxyethyl)phen- yl} ethynyl]benzoic
acid and 4-[{3-(1-adamantyl)-4-(3-hydroxypropyl)phenyl-
}ethynyl]benzoic acid.
[0025] Preference is given, in the present invention, to using
agonist ligands which are specific for receptors of the RAR-.gamma.
type which exhibit a ratio R which is greater than or equal to 50.
As such, preference is given to using
6-3-(1-adamantyl)-4-hydroxyphenyl)-2-naphtha- noic acid,
6-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalene-2-carbonyl-
)-naphthalene-2-carboxylic acid,
6-(1,2,3,4-tetrahydro-1,4a,9b-trimethyl-1-
,4-methanodibenzofuran-8-yl)-naphthalene-2-carboxylic acid,
6-[hydroxyimino-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)me-
thyl]naphthalene-2-carboxylic acid,
5-(5,5,8,8-tetramethyl-5,6,7,8-tetrahy-
droanthracen-2-yl)thiophene-2-carboxylic acid,
(-)-6-[hydroxy-(5,5,8,8-tet-
ramethyl-5,6,7,8-tetrahydronaphthalen-2-yl)methyl]naphthalene-2-carboxylic
acid, 6-[3-(1-adamantyl-4-(2-hydroxypropyl)phenyl]-2-naphthoic
acid, 6-[3-(1-adamantyl-4-(2,3-dihydroxypropyl)phenyl]-2-naphthoic
acid, 4-[3-(1-adamantyl)-4-hydroxyphenylethynyl]benzoic acid,
5-[3-(1-adamantyl)-4-hydroxyphenylethynyl]-2-thiophenecarboxylic
acid,
5-[3-(1-adamantyl)-4-methoxyphenylethynyl]-2-thiophenecarboxylic
acid,
6-[3-(1-adamantyl)-4-(3-methoxy-2-hydroxypropyl)phenyl]-2-naphthoic
acid,
1-methyl-4-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydroanthracen-2-yl)-1H-pyrr-
ole-2-carboxylic acid,
(-)-2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5-
,6,7,8-tetrahydronaphthalen-2-yl)prop-1-ynyl]benzoic acid and
2-hydroxy-4-[3-hydroxy-3-(5,5,8,8-tetramethyl-5,6,7,8-tetrahydro-naphthal-
en-2-yl)but-1-ynyl)benzoic acid.
[0026] 6-3-(1-Adamantyl)-4-hydroxyphenyl)-2-naphthanoic acid is
particularly preferred.
[0027] Thus, it will be possible to use the pharmaceutical
composition comprising the agonist ligand which is specific for
receptors of the RAR-.gamma. type when it is necessary to increase
the rate of apoptosis. Naturally, this effect will only be achieved
in cell populations in which apoptosis can be induced by activating
receptors of the RAR-.gamma. type and therefore, in particular, in
which receptors of the RAR-.gamma. type are present, as is the
case, more specifically, in cells derived from the thymus.
[0028] It may prove to be necessary to increase the rate of
apoptosis in two cases in the main. The first case relates to
diseases or disorders which are linked to an inadequate rate of
apoptosis. The second case relates to treatments which are required
during a transplantation in order to decrease the effects tending
to reject the transplanted organ. Thus, it is possible to consider
increasing immunotolerance to the transplanted organ by decreasing
the immune response of the T cells by means of inducing their rate
of apoptosis.
[0029] Diseases or disorders which are linked to an inadequate rate
of apoptosis, and which may more specifically be mentioned, are
disorders which are due to precancerous conditions or cancers which
can be the consequence of a proliferation of certain cell
populations, autoimmune diseases, allergies or inflammatory
reactions in which the number of cells causing damage is observed
to be too high, or else in certain viral infections in which
proteins of the virus have an antiapoptotic effect. Thus, in the
case of autoimmune diseases, more specific mention may be made of
insulin-dependent diabetes, active chronic hepatitis, rheumatoid
arthritis, pemphigus, multiple sclerosis, myasthenia, systemic
lupus erythematosus, Crohn's disease and psoriasis. Actinic
keratosis may be mentioned with regard to precancerous conditions.
In the case of cancer, more specific mention may be made of
lymphomas, carcinomas, such as cancer in the ENT sphere, and
hormone-dependent tumours, such as ovarian cancers. Of the
abovementioned viral infections, those which may in particular be
cited are herpes viruses, adenoviruses and variola viruses (pox
viruses). Allergies or inflammatory reactions which may be
mentioned are contact eczema, atopic eczema, asthma and
urticarias.
[0030] The composition according to the invention can be
administered by the enteral, parenteral, topical or ocular route.
Preference is given to packaging the pharmaceutical composition in
a form which is suitable for administration by the systemic route
(for injection or perfusion).
[0031] When administered by the enteral route, the composition,
more specifically the pharmaceutical composition, can be in the
form of tablets, hard gelatin capsules, coated tablets, syrups,
suspensions, solutions, powders, granules, emulsions, microspheres
or nanospheres, or lipid or polymeric vesicles which permit a
controlled release. When administered by the parenteral route, the
composition may be in the form of solutions or suspensions for
perfusion or injection.
[0032] The agonist ligands which are specific for receptors of the
RAR-.gamma. type and which are used in accordance with the
invention are generally administered in a daily dose of from 0.01
mg/kg to 100 mg/kg of body weight, with the dose being given in
from 1 to 3 administrations.
[0033] When administered by the topical route, the pharmaceutical
or cosmetic composition according to the invention is more
specifically intended for treating the skin and the mucous
membranes and can be in the form of ointments, creams, milks,
pomades, powders, imbibed buffers, solutions, gels, sprays, lotions
or suspensions. It may also be in the form of microspheres or
nanospheres, or lipid or polymeric vesicles, or of polymeric
patches and hydrogels, which permit a controlled release. This
composition, which is administered by the topical route, can be
present either in anhydrous form or in aqueous form.
[0034] When administered by the ocular route, the composition is
chiefly in the form of eye drops.
[0035] The agonist ligands which are specific for receptors of the
RAR-.gamma. type are used by the topical or ocular route at a
concentration which is generally between 0.001% and 10% by weight,
preferably between 0.1 and 1% by weight, based on the total weight
of the composition.
[0036] The present invention finally relates to a process for
preventing and/or combatting photoinduced or chronological ageing
of the skin, characterized in that a cosmetic composition which
induces apoptosis such as previously described, that is to say
which comprises at least one agonist ligand which is specific for
receptors of the RAR-.gamma. type in a cosmetically acceptable
medium, is applied to the skin.
[0037] Naturally, the compositions as previously described may
additionally comprise inert or even pharmacodynamically active
additives or combinations of these additives, in particular:
wetting agents; depigmenting agents such as hydroquinone, azelaic
acid, caffeic acid or kojic acid; emollients; moisturizing agents
such as glycerol, PEG 400, thiamorpholinone and its derivatives, or
urea; antiseborrhoeic agents or anti-acne agents, such as
S-carboxymethylcysteine, S-benzyl-cysteamine, and their salts or
derivatives, or benzoyl peroxide; antifungal agents such as
ketoconazole or the polymethylene-4,5-iso-thioazolidonin-3-ones;
antibacterial agents, carotenoids and, in particular,
.beta.-carotene; antipsoriatic agents such as anthralin and its
derivatives; and, finally, eicosa-5,8,11,14-tetraynoic and
eicosa-5,8,11-trynoic acids, and their esters and amides.
[0038] These compositions may also comprise taste-improving agents,
preservatives such as esters of parahydroxybenzoic acid,
stabilizers, moisture-regulating agents, pH-regulating agents,
agents for modifying the osmotic pressure, emulsifying agents, UV-A
and UV-B filters, antioxidants, such as .alpha.-tocopherol,
butylhydroxyanisole or butylhydroxytoluene.
[0039] Naturally, the skilled person will ensure that the possible
compound(s) to be added to these compositions is/are selected such
that the advantageous properties which are intrinsically attached
to the present invention are not altered, or are not substantially
altered, by the addition which is envisaged.
[0040] Several examples which are intended to illustrate the
present invention, but which are in no way limiting, will now be
given.
EXAMPLE 1
[0041] This experiment demonstrates the in vivo efficacy of an
agonist ligand which is specific for RAR-.gamma. as an inducer of
apoptosis.
[0042] 4-week-old male NMRI mice (sold by LATI, Godollo, Hungary)
were used. In order to induce apoptosis in the thymus, these male
mice were treated by a single injection with either 0.5 mg of
dexamethasone, or 0.5 mg of
6-3-(1-adamantyl)-4-hydroxyphenyl)-2-naphthanoic acid, dissolved in
a mixture of 0.8 ml of physiological saline and 0.2 ml of
ethanol.
[0043] Dexamethasone is a well-known inducer of apoptosis. Thus,
involution of the thymus is observed: i.e. a decrease of
approximately 75% in the weight of the thymus at 48 hours after the
abovementioned treatment. A similar effect is observed with
6-3-(1-adamantyl)-4-hydroxyp- henyl)-2-naphthanoic acid, with in
this case a decrease of approximately 60% in the weight of the
thymus being observed at 48 hours after treatment.
[0044] In addition, in a similar manner to this test, thymus
samples were removed from untreated or treated animals at different
times after the treatment. After washing and homogenization, their
transglutaminase activity was determined by detecting the
incorporation of [.sup.3H]putrescine into N,N'-dimethylcasein. The
activity of the transglutaminase is calculated in nmoles of
[.sup.3H]putrescine incorporated into the protein per hour.
[0045] Tissue transglutaminase has been reported to be one of the
elements involved in bringing about apoptosis [Piacentini, M. et
al. (1994) Apoptosis: The Molecular Basis of Apoptosis in Disease.
Curr. Comm. in Cell & Mol. .sctn.Biol. 8 (Tomei L. D. and Cope,
F. O. ed) pp. 143-165].
[0046] Thus, it is observed that the above-described involution of
the thymus results from treating the mouse with a
RAR-.gamma.-specific agonist ligand and is an event which is
concomitant with, and proportional to, the activity of the tissue
transglutaminase.
[0047] Furthermore, this involution of the thymus is correlated
with the appearance of DNA fragments following the analysis, by
agarose gel electrophoresis, of the DNA which is recovered from
this treated thymus (details of the procedure are given in Example
2).
EXAMPLE 2
[0048] This example demonstrates the in vitro efficacy of a
RAR-.gamma.-specific agonist ligand as an inducer of apoptosis as
compared with other types of retinoid.
Culture and Preparation of the Cells
[0049] Thymocyte suspensions are prepared from the thymus glands of
untreated four-week-old male NMRI mice (sold by LATI, Godollo,
Hungary). The medium employed is Sigma RPMI 1640 medium which is
supplemented with Gibco foetal calf serum, 2 mM glutamine and 100
IU of penicillin and 100 .mu.g of streptomycin/ml. The thymocytes
are then washed, and diluted in order to obtain a final
concentration of 10.sup.7 cells/ml, before being incubated at
37.degree. C. in a humidified incubator under an atmosphere of 5%
CO.sub.2 and 95% air. Death of the cells is measured by the uptake
of trypan blue.
Qualitative and Quantitative Analysis of the DNA
[0050] The thymocytes are incubated in 24 wells with various
compounds to be tested at different concentrations. After 6 hours
of incubation, 0.8 ml of the cell suspensions was lysed by adding
0.7 ml of lysis buffer containing 0.5% (v/v) Triton X-100, 10 mM
Tris, 20 mM EDTA, pH 8.0, before being centrifuged at 13,000 g for
15 minutes.
[0051] Quantitative analysis of the DNA: the DNA contained in the
supernatant (the fragments) and the pellet (intact chromatin) was
precipitated with an equivalent quantity of 10% trichloroacetic
acid, resuspended in 5% trichloroacetic acid and then quantified
using the diphenylamine reagent (Burton, K. (1956) Biochem. J., 62,
315-322).
[0052] Qualitative analysis of the DNA: in parallel, the
supernatant was precipitated overnight in ethanol containing 0.15
mM NaCl. The pellets are redissolved in a buffer containing 10 mM
Tris, 1 mM EDTA, pH 8.0, and these solutions are then treated with
RNase; they are then sequentially extracted with an equal volume of
phenol and chloroform/isoamyl alcohol (24/1), after which the DNA
is precipitated in ethanol before being electrophoresed for 3 hours
at 60 V in a 1.8% agarose gel. The DNA fragments were then
visualized with UV light after the gel had been stained with
ethidium bromide. The gels which are obtained present the picture
of a ladder of DNA fragments which are multiples of from 180 to 200
base pairs and which are typical of an apoptosis induction.
Throughout the experiments, the degree of fragmentation correlates
with the number of cells which are dead and positive to the trypan
blue test.
[0053] The results of the quantitative analysis are assembled in
Table 1 below.
1TABLE 1 Quantities of RAR-.alpha. RAR-.gamma. compounds % of DNA
Compounds Kd Kd R (nM) fragments ATRA 15.5 3 5.16 10.sup.-8 2
10.sup.-6 4 9-cisRA 7 17 0.41 10.sup.-8 4 10.sup.-6 15 CD437 6500
77 84.42 10.sup.-8 12 10.sup.-6 22 CD666 2240 68 32.94 10.sup.-8 10
10.sup.-6 20 CD2325 1144 53 21.58 10.sup.-8 5 10.sup.-6 20 CD2019
1100 160 6.87 10.sup.-8 0 10.sup.-6 15 ATRA is all-trans retinoic
acid 9-cisRA is 9-cis-retinoic acid CD437 is
6-3-(1-adamantyl)-4-hydroxyphenyl)-2-naphthanoic acid, CD666 is
(E)-4-(1-hydroxy-1-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthyl)-2--
propenyl)benzoic acid, CD2325 is 4-[(E)-2-(3-(1-adamantyl)-4-hydro-
xyphenyl)-1-propenyl]benzoic acid, CD2019 is
6-(3-(1-methylcyclohexyl)-4-methoxyphenyl)-2-naphthanoic acid
[0054] ATRA is all-trans retinoic acid
[0055] 9-cisRA is 9-cis-retinoic acid
[0056] CD437 is 6-3-(1-adamantyl)-4-hydroxyphenyl)-2-naphthanoic
acid,
[0057] CD666 is
(E)-4-(1-hydroxy-1-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-
-2-naphthyl)-2-propenyl)benzoic acid,
[0058] CD2325 is
4-[(E)-2-(3-(1-adamantyl)-4-hydroxyphenyl)1-propenyl]benz- oic
acid,
[0059] CD2019 is
6-(3-(1-methylcyclohexyl)-4-methoxyphenyl)2-naphthanoic acid
[0060] The percentage of DNA fragments in this table corresponds to
the difference between the percentage of DNA fragments obtained in
treated thymocytes and the percentage of DNA fragments obtained in
nontreated thymocytes (basal rate of apoptosis for these
thymocytes).
[0061] These results demonstrate that the percentage appearance of
DNA fragments increases as R increases. Thus, the
apoptosis-inducing effect increases as the specificity of the
ligand for RAR-.gamma. increases.
* * * * *