U.S. patent application number 12/884820 was filed with the patent office on 2011-01-13 for pharmaceutical or cosmetic composition and use of a pkc inhibitor with an mmp inhibitor for inhibiting langerhans' cell migration.
This patent application is currently assigned to Laboratories Expanscience. Invention is credited to Philippe MSIKA, Nathalie PICCARDI, Antoine PICCIRILLI.
Application Number | 20110009333 12/884820 |
Document ID | / |
Family ID | 8856405 |
Filed Date | 2011-01-13 |
United States Patent
Application |
20110009333 |
Kind Code |
A1 |
MSIKA; Philippe ; et
al. |
January 13, 2011 |
PHARMACEUTICAL OR COSMETIC COMPOSITION AND USE OF A PKC INHIBITOR
WITH AN MMP INHIBITOR FOR INHIBITING LANGERHANS' CELL MIGRATION
Abstract
Disorders of the skin or mucous membranes which are linked to
migration of Langerhans cells are treated by the administration of
an effective amount of at least one retinoid, optionally in
combination with a PKC inhibitor.
Inventors: |
MSIKA; Philippe; (Paris,
FR) ; PICCARDI; Nathalie; (Saint Egreve, FR) ;
PICCIRILLI; Antoine; (Epernon, FR) |
Correspondence
Address: |
FOLEY AND LARDNER LLP;SUITE 500
3000 K STREET NW
WASHINGTON
DC
20007
US
|
Assignee: |
Laboratories Expanscience
|
Family ID: |
8856405 |
Appl. No.: |
12/884820 |
Filed: |
September 17, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12073220 |
Mar 3, 2008 |
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12884820 |
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10416464 |
Aug 25, 2003 |
7511010 |
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PCT/FR01/03568 |
Nov 14, 2001 |
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12073220 |
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Current U.S.
Class: |
514/18.7 ;
514/336; 514/529; 514/557; 514/563; 514/569; 514/613; 514/703;
514/725 |
Current CPC
Class: |
A61K 31/13 20130101;
A61K 8/41 20130101; A61K 31/56 20130101; A61P 29/00 20180101; A61K
31/045 20130101; A61P 37/08 20180101; A61K 2800/75 20130101; A61P
17/00 20180101; A61K 31/13 20130101; A61Q 19/005 20130101; A61Q
19/00 20130101; A61K 8/68 20130101; A61P 37/06 20180101; A61K
2300/00 20130101; A61K 36/48 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 36/48 20130101; A61K 31/56 20130101; A61P
17/06 20180101; A61P 43/00 20180101 |
Class at
Publication: |
514/18.7 ;
514/725; 514/703; 514/557; 514/529; 514/613; 514/569; 514/336;
514/563 |
International
Class: |
A61K 38/02 20060101
A61K038/02; A61P 17/00 20060101 A61P017/00; A61P 29/00 20060101
A61P029/00; A61P 37/06 20060101 A61P037/06; A61K 31/07 20060101
A61K031/07; A61K 31/11 20060101 A61K031/11; A61K 31/203 20060101
A61K031/203; A61K 31/215 20060101 A61K031/215; A61K 31/16 20060101
A61K031/16; A61K 31/192 20060101 A61K031/192; A61K 31/4436 20060101
A61K031/4436; A61K 31/196 20060101 A61K031/196 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 14, 2000 |
FR |
0014607 |
Claims
1.-11. (canceled)
12. A method for the treatment of disorders of the skin or mucous
membranes linked to Langerhans cell migration comprising the
administration, to a person in need thereof, of an effective amount
of a composition comprising at least one retinoid.
13. The method of claim 12, wherein said disorder is selected from
the group consisting of allergic reactions, atopic eczema, contact
eczema, skin sensibility, skin reactivity, inflammatory dermatoses,
irritant dermatitis, transplant rejection, autoimmune diseases,
inflammatory diseases, psoriasis and photoimmunosuppression.
14. The method of claim 12, wherein said retinoid is selected from
the group consisting of nonaromatic retinoids, vitamin A,
monoaromatic retinoids and polyaromatic retinoids.
15. The method of claim 14, wherein said nonaromatic retinoid is
selected from the group consisting of retinaldehyde, tretinoin and
9-cis-retinoic acid.
16. The method of claim 14, wherein said monoaromatic retinoid is
selected from the group consisting of etretinade,
all-trans-acitretin and motrerinide.
17. The method of claim 14, wherein said polyaromatic retinoid is
selected from the group consisting of adapalene, tazarotene,
tamibarotere and arotinoid methyl sulfone.
18. The method of claim 12, wherein the composition further
comprises a PKC-inhibitor selected from the group consisting of
peptide extracts of lupin, sphingolipids and ceramids.
19. The method of claim 18, wherein the sphingolipid is selected
from the group consisting of sphingosine, phytosphingosine and
isomers thereof.
20. The method of claim 18, wherein the peptide extract of lupin is
LU105.
21. The method of claim 12, wherein the retinoid concentration in
the composition is between approximately 0.001 and approximately
10% by weight, relative to the total weight of the composition.
22. A method for the treatment of disorders of the skin or mucous
membranes linked to Langerhans cell migration comprising the
administration, to a person in need thereof of an effective amount
of a composition comprising at least one retinoid selected from the
group consisting of vitamin A and 9-cis-retinoic acid.
23. The method of claim 22, wherein the composition further
comprises a PKC-inhibitor selected from the group consisting of
peptide extracts of lupin, sphingolipids and ceramids.
Description
[0001] The present invention relates to a pharmaceutical or
cosmetic composition, and also to the use of at least one active
compound for inhibiting the migration of cells involved in the
immune and/or inflammatory and/or irritative response, such as
dermal dendrocytes, monocytes, lymphocytes, and more particularly
Langerhans cells.
[0002] One of the main functions of the skin is to protect the body
against attacks from the outside environment. This protection is
provided in large part through the cooperation of cells present in
the skin, these being cells which are capable, in the presence of a
harmful agent, of generating an inflammatory and/or immune response
directed against the harmful agent. They are dendritic cells,
epidermal Langerhans cells (LCs) and dermal dendrocytes, monocytes,
lymphocytes, keratinocytes, mastocytes and vascular endothelial
cells.
[0003] LCs are dendritic cells derived from the spinal cord which
resides in the nonlymphoid tissues, such as the skin and the mucous
membranes (mouth, lungs, bladder, rectum, vagina). In the skin, the
LCs intercalate between the epidermal keratinocytes in the
suprabasal position. In terms of ultrastructure, they are
characterized by the presence of a specific organelle of membrane
origin, the Birbeck granule. In immunohistochemical terms, LCs
express in particular the CD1a molecule and the class II Major
Histocompatibility Complex molecules.
[0004] LCs play a determining role in immunity, as
antigen-presenting cells. Specifically, experiments carried out in
mice demonstrate that LCs capture the antigens present in the
epidermis and migrate toward the lymphoid tissues draining the
skin, where they present the antigen to T cells. Initiation of the
immune response to the skin depends on the ability of the LCs to
leave the epidermis in order to migrate to the proximal lymph
nodes. Various factors can influence this migration: the expression
of adhesion molecules, the extracellular matrix proteins, haptens,
cytokines, etc. However, the mechanisms involved in LC migration
are not yet entirely elucidated. In particular, before reaching the
lymph nodes, LCs must not only cross the dermoepidermal junction
(DEJ), but also make themselves a path through the dermal
extracellular matrix (ECM). The DEJ is mainly composed of laminin
5, of type IV and VII collagen, of nidogen and of perlecan. The ECM
which surrounds the dermal fibroblasts essentially contains type I
and III collagens.
[0005] Pathological conditions of the dermatological type can also
be observed as resulting from LC migration subsequent to capture of
a surface antigen.
[0006] In atopic eczema, the LCs are capable of attaching IgEs at
the surface and of inducing: a pathological immune response.
[0007] In contact eczema, the LCs play a central role since they
capture and process the antigen before presenting it to T
lymphocytes. This T lymphocyte will keep it in the memory and the
immune reaction will be triggered at the second contact.
[0008] Given the above, it was therefore highly desirable to be
able to modify the migratory ability of LCs, in order to attempt to
modify the induction of the immune and/or inflammatory
reaction.
[0009] It has now been found, entirely surprisingly and
unexpectedly, that some compounds make it possible to spectacularly
inhibit Langerhans cell migration induced by the presence of an
allergenic agent.
[0010] The present invention thus relates to a pharmaceutical or
cosmetic composition, characterized in that it comprises at least
one active compound for inhibiting Langerhans cell migration, said
active compound being chosen from the group consisting of protein
kinase C(PKC) inhibitors, matrix metalloprotease (MMP) inhibiting
compounds, and combinations thereof, and at least one
pharmaceutically or cosmetically acceptable excipient.
[0011] According to the invention, the term "protein kinases C" or
"PKCs" is intended to mean the enzymes which catalyze a
phosphorylation reaction on the cell substrate.
[0012] When they are activated, PKCs phosphorylate specific serine
or threonine residues on protein substrates, which vary according
to cell type. In many cells, PKC activation increases the
transcription of specific genes.
[0013] Protein kinases C(PKCs) are proteins encoded by a family of
genes (11 different isoforms). It is in particular known that these
proteins are involved in the extracellular signal transduction
mediated by growth factors and cytokines, and also by a certain
number of other biological molecules. Protein kinase
.beta.2(PKC-.beta.2) appears to be expressed specifically by
epidermal LCs.
[0014] Any compound known to those skilled in the art to inhibit
the phosphorylation activity of PKCs can thus be used as a
PKC-inhibiting compound according to the present invention. Mention
may, for example, be made of the polypeptides described in
application WO 99/43805 (Incyte Pharma Inc.).
[0015] In particular, the active compound is a PKC-inhibiting
compound chosen from the group consisting of nonspecific PKC
inhibitors, inhibitors specific for the isoform PKC-.beta.2, and
combinations thereof.
[0016] More particularly, the active compound is a PKC-inhibiting
compound chosen from the group consisting of phenol and polyphenol
compounds, procyanidins (catechins, epicatechins, etc.),
alphaamyrin, lupeol, lupeol linoleate, sterols, stanols,
triterpenic alcohols and hydrogenated homologs thereof, antibiotics
such as staurosporin or Ro-318425 (or
2-(8)-(aminomethyl)-6,7,8,9-tetrahydropyridol(1,2-a)-indol-3-yl)-3-(1-met-
hylindol-3-ylmaleimide, HCl) as marketed by the company Calbiochem,
compounds which act by competition with physiological PKC
activators, such as diacylglycerol or phorbol ester, cutaneous
lipids of the (lyso)sphingolipid type, lysophospholipids such as
ceramides and pseudoceramides, or sphingosine and phytosphingosine,
and the derivatives, precursors, analogs and homologs of these
compounds, of natural or synthetic origin.
[0017] According to the invention, the term "phenol and polyphenol
compounds" is intended to mean simple phenols, benzoquinones,
phenolic acids, acetophenones, phenylacetic acids, hydroxycinnamic
acids, coumarins and isocoumarins, chromones, naphthoquinones,
xanthones, anthraquinones, flavonoids, lignans and neolignans,
lignins, chalcones, dihydrochalcones, aurones, flavones, flavonols,
dihydroflavonols, flavanones, flavanols, flavandiols or
leucoanthocyanidins, anthocyanidins, isoflavonoids, biflavonoids,
proanthocyanidins: and tannins which are condensed.
[0018] According to the invention, the term "sterols" is more
particularly intended to mean sterol, i.e. the
perhydro-1,2-cyclopentanophenanthrene compound having a hydroxyl
group at position 3, and the analogs of sterol of general formula
(I) below.
[0019] Thus, preferably, the sterols which can be used according to
the invention correspond to the general formula:
##STR00001##
in which the unsaturation represented as a dotted line at position
5 corresponds to the unsaturation in the case of sterols, R
represents a linear or branched hydrocarbon-based chain which may
or may not be unsaturated, comprising from 1 to 25 carbon atoms. In
particular, R is chosen from the group consisting of
C.sub.1-C.sub.12 alkyl groups, C.sub.1-C.sub.8 alkoxy groups,
C.sub.2-C.sub.8 alkenyl groups, C.sub.2-C.sub.8 alkynyl groups,
C.sub.3-C.sub.8 cycloalkyl groups, halogenated C.sub.2-C.sub.8
alkenyl groups, and halogenated C.sub.2-C.sub.8 alkynyl groups. The
term "halogenated" denotes one or more halogen substituents, namely
one or more chlorine, fluorine, bromine or iodine atom(s).
[0020] Among the sterols which may advantageously be used according
to the invention, mention may in particular be made of
.beta.-sitosterol, .alpha.-sitosterol, .gamma.-sitosterol,
stigmasterol or, alternatively, campesterol, and mixtures thereof.
For example, .beta.-sitosterol can be used in the form of the
product known as "Ultra" (mainly comprising .beta.-sitosterol) as
marketed by the company Kaukas. In the case of use of a mixture of
sterols, mention may, for example, be made of the product known as
"Generol" comprising mainly .beta.-sitosterol (approximately 50% by
weight), stigmasterol and campesterol, as marketed by the company
Henkel or else the product "Primal" from the company Kaukas.
[0021] Among the triterpenic alcohols which may advantageously be
used according to the invention, mention may in particular be made
of .beta.-amyrin, erythrodiol, taraxasterol, cycloartenol,
24-methylene-cycloartanol, lupeol, lanosterol and mixtures
thereof.
[0022] According to the invention, the term "hydrogenated homologs"
of a triterpenic alcohol is intended to mean the corresponding
triterpenic alcohol compound(s) in which the unsaturated bond(s)
possibly present has (have) been hydrogenated (i.e. converted to a
saturated bond) according to methods well known to those skilled in
the art.
[0023] Even more particularly, the active compound is a
PKC-inhibiting compound corresponding to the general formula
##STR00002##
in which:
[0024] R.sub.1 represents a hydrogen atom or a linear or branched
hydrocarbon-based chain having from 1 to 40 carbon atoms, in
particular from 8 to 36, and more particularly from 16 to 20 carbon
atoms, which may comprise one or more double bonds and which may
comprise one or more hydroxyl substituents;
[0025] R.sub.2 represents a hydrogen atom or a group of formula
Z--CO-- where Z represents: [0026] (u) a linear or branched
hydrocarbon-based chain having from 1 to 40 carbon atoms, in
particular from 16 to 36, and more particularly from 20 to 34
carbon atoms, which may comprise one or more double bonds and which
may comprise one or more hydroxyl substituents; or [0027] (v) a
group R.sub.6--CO--O-A- where R.sub.6 represents a linear or
branched hydrocarbon-based chain having from 1 to 40 carbon atoms,
in particular from 8 to 38, and more particularly from 18 to 34
carbon atoms, which may comprise one or more double bonds and which
may comprise one or more hydroxyl substituents, and where A
represents a linear or branched hydrocarbon-based chain having from
1 to 40 carbon atoms, in particular from 16 to 38, and more
particularly from 24 to 36 carbon atoms, which may comprise one or
more double bonds and which may comprise one or more hydroxyl
substituents;
[0028] X represents a hydrogen atom, a monosaccharide residue or
oligosaccharide residue, in particular a galactose residue, the
sulfogalactose group, phosphorylcholine or the group of formula
(GalNAc)(Sia)Gal-Glc-; and
[0029] Y represents a hydrogen atom or a group of formula
R.sub.3--W--CHOH-- where [0030] R.sub.3 represents a linear or
branched hydrocarbon-based chain having from 1 to 40 carbon atoms,
in particular from 8 to 36, and more particularly from 14 to 18
carbon atoms, which may comprise one or more double bonds; and
where [0031] W represents: [0032] (i) a group of formula
--CH.dbd.CH--; [0033] (ii) a group of formula
--CH.sub.2--CH(OR.sub.4)-- where R.sub.4 represents a hydrogen atom
or a group R.sub.5--CO-- with R.sub.5 representing a linear or
branched hydrocarbon-based chain having from 1 to 40 carbon atoms,
in particular from 8 to 36, and more particularly from 14 to 18
carbon atoms, which may comprise one or more double bonds and which
may comprise one or more hydroxyl substituents; or [0034] (iii) a
group --CH.sub.2--CH.sub.2.
[0035] Even more particularly, the active compound is a
PKC-inhibiting compound chosen from the group consisting of
sphingolipids and lysophospholipids, such as those cited in the
table below:
TABLE-US-00001 TABLE 1 Sphingolipid and lysosphingolipid structures
SPHINGOLIPIDS LYSOSPHINGOLIPIDS Structure ##STR00003## ##STR00004##
X: H-- Ceramide Sphingosine X: Galactose- Galactocerebroside
Psychosine (Galactosylsphingosine) X: Sulfogalactose- Sulfatide
Lysosulfatide (Sulfogalactosylsphingosine) ##STR00005## GM.sub.2
Lyso GM.sub.2 X: Phosphorylcholine- Sphingornyelin
Lysosphingomyelin
[0036] As a PKC-inhibitor compound, mention may also, more
particularly, be made of cutaneous lipids of the sphingolipid or
lysophospholipid type.
[0037] As sphingolipids, mention may be made of those among the
most elementary, such as sphingosine
(D-erythro-1,3-dihydroxy-2-amino-4-trans-octadecene) and isomers
thereof, or phytosphingosine
(D-ribo-1,3,4-trihydroxy-2-aminooctadecane) and isomers thereof;
but also, lysosphingolipids (including lysosulfatide and
psychosine), sulfogalactosylsphingosine, sphinganine
(2-amino-1,3-octadecanediol) and sphingomyelins.
[0038] As phospholipids, mention may be made of those among the
families of phosphatidylamino alcohols and phosphatidyl polyols.
The phosphatidylamino alcohol group comprises in particular
phosphatidylethanolamines (or phosphatidylcolamines),
phosphatidylcholines, phosphatidylserines and
N-acylphosphatidylethanol-amines. As regards the phosphatidyl
polyol group, it comprises phosphatidylcholinositols,
diphosphoinositides, lysodiphosphoinositides, phosphatidyl
glycerols and cardiolipids.
[0039] As a PKC-inhibiting compound, mention may also more
particularly be made of ceramides, in particular the ceramides of
the intercorneocyte cement of the epidermis and also the ceramide
precursors, mainly sphingosine and phytosphingosine.
[0040] In general, the ceramides can be synthesized chemically
(reference is in particular made to pseudoceramides), may be of
animal origin (relatively high concentrations of sphingolipids are
present in the mammalian brain and vertebral column), may be of
plant origin (mainly cerebrosides and other glycosylated
sphingolipids) or else may be derived from yeast (stereochemical
configuration identical to that of the ceramides naturally present
in human skin).
[0041] The ceramides of the intercorneocyte cement of the epidermis
can be separated using conventional methods (thin layer
chromatography) into six fractions, corresponding to compounds
which differ by the nature of the fatty acids and the nature of the
base involved (sphingosines, which are unsaturated, or
phytosphingosines, which are saturated). Table 2 below illustrates
the respective structures present in these fractions, according to
the classification of Werts and Downing. Fraction 6 can, itself, be
subdivided, by more refined methods, into two entities: ceramides
6a and 6b.
TABLE-US-00002 TABLE 2 The six main fractions of epidermal
ceramides ##STR00006## ##STR00007## ##STR00008## ##STR00009##
##STR00010## ##STR00011## ##STR00012##
[0042] Thus, the ceramides 1, the least polar, comprise an entirely
specific structure which is found in ceramide 6a: a long-chain
omega-hydroxy acid amidating the base, and attached, at its omega
end, via an ester bond, to another fatty acid (O-acylceramides). In
the case of fraction 1, the fatty acids linked to the sphingosine
are essentially C24, C26, C30, C32 or C34, which can be saturated
(as represented in FIG. 5 for a C30), monoethylenic (mainly for
C30, C32 and C34) or diethylenic (C32 and especially C34). As
regards the fatty acid attached to the omega end of the preceding
one, it is, largely predominantly for the ceramides 1, linoleic
acid; the essential role in the hybrid barrier function of the
epidermis is well known.
[0043] Fraction 2, which has a more conventional structure
(sphingosines or dihydrosphingosines linked, via an amide bond, to
a fatty acid, mainly C20 to C28), is the most abundant.
[0044] Fraction 3 is quite similar, the difference relating to the
nature of the base, which, in this case, is essentially represented
by saturated phytosphingosines.
[0045] Fractions 4 and 5 are essentially characterized by the
presence of alpha-hydroxy acids linked to a sphingosine.
[0046] Fraction 6b is close to fractions 4 and 5, comprising an
alpha-hydroxy acid, but linked to a saturated phytosphingosine.
[0047] Fraction 6a, like the ceramide 1, comprises the
characteristic motif which is found only in epidermal ceramides,
i.e. the ester bond between the hydroxyl in the omega-position of a
fatty acid linked to a sphingosine, and the carboxylic group of a
terminal fatty acid which, this time, is not linoleic acid, but an
alpha-hydroxy acid.
[0048] Phytoceramides (ceramides based on phytosphingosine),
synthetic cholesterol ceramides, and galacto- or glucocerebrosides
should also be mentioned.
[0049] Finally, among the PKC-inhibiting compounds which can be
used according to the present invention, sphingosine is present
naturally in the skin and plays, inter alia, an important role in
the barrier function of the stratum corneum, as a precursor of
sphingolipids (ceramides and sphingoglycolipids). It may be derived
from a biological source, such as extracts of bovine brains, or via
the synthetic pathway, using serine, as described, for example, in
the article by Newman, J. Am. CHEM., 95 (12): 4098 (1973). Mention
may more particularly be made of the isomeric forms of sphingosine:
D-erythro, L-threo, L-erythro and D-threo. The D-erythro form is
the form most commonly present in nature.
[0050] According to the present invention, the PKC-inhibiting
compounds which can be used, as active compounds for inhibiting
Langerhans cell migration, comprise the isomers, the derivatives
(salts, complexes, etc.), the analogs, the homologs, the precursors
and the metabolites of the PKC-inhibiting compounds described
above.
[0051] According to the invention, the expression "matrix
metalloprotease (MMP) inhibiting compounds" is intended to mean any
compound known to those skilled in the art for its ability to
inhibit the activity of extracellular matrix degradation by
MMPs.
[0052] MMPs constitute a family of zinc-dependent enzymes
(currently more than about twenty have been identified and
characterized) which have a very conserved structure and which
possess the ability to degrade the components of the extracellular
matrix. They are classified, depending on the nature of their
substrate, as collagenases, gelatinases and stromelysin. They can
be synthesized by various cell types in the skin (fibroblasts,
keratinocytes, macrophages, endothelial cells, eosinophils,
Langerhans cells, etc.). The MMP group thus consists of four
subclasses: (1) collagenases, (2) gelatinases, (3) stromelysins and
(4) membrane-type MMPs (MT-MMPs). The activity of MMPs can be
modulated by naturally present protease inhibitors, such as tissue
inhibitors of metalloproteases (TIMPs; in particular TIMP-1 and
TIMP-2).
[0053] The predominant role of MMPs in the proteolytic remodeling
of the extracellular matrix is now clearly established, both in
physiological situations (cicatrization, angiogenesis, embryonic
development, etc.) and pathological situations (chronic ulcer,
photo-induced aging of the skin, tumor cell invasion, etc.).
[0054] In particular, the active compound for inhibiting Langerhans
cell migration is a compound which inhibits at least one MMP chosen
from the group consisting of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9,
MMP-13 and MMP-18.
[0055] According to the present invention, the expression
"MMP-inhibiting compound", as an active compound for inhibiting
Langerhans cell migration, is intended in particular to mean tissue
inhibitors of metalloproteases (TIMPs), alpha-2-macroglobulin,
plasminogen activator inhibitors, zinc chelators, bryostatin-1,
antibiotics (doxycyclins, minocycline, etc.), synthetic or natural
peptides having a structure similar to MMP substrates (batimastat,
marimastat, etc.), retinoids (in particular nonaromatic retinoids
such as retinaldehyde, tretinoin or 9-cis-retinoic acid, vitamin A,
monoaromatic retinoids such as etretinate, all-trans-acitretin or
motrerinide, and polyaromatic retinoids such as adapalene,
tazarotene, tamibarotene or arotinoid methyl sulfone), antioxidants
(singlet oxygen scavengers, etc.), anticancer agents (or
"antimetastatics"), malt hydrolysates such as Colalift marketed by
the company Coletica, extracts of marine algae such as Kelpadelie
marketed by the company Secma, extracts of shark cartilage such as
the MDI complex marketed by the company Atrium, rice peptides such
as, for example, Colhibin marketed by the company Pentapharm, and
peptide extracts of lupin.
[0056] More particularly, the MMP-inhibiting compound according to
the present invention is chosen from the group consisting of the
peptide extracts of lupin, or "lupin peptides", such as those
described in patent application FR-99 04 875 filed on Apr. 19,
1999, in the name of the company Laboratoires Pharmascience.
Mention may in particular be made of the peptide extract described
in application FR 99 04875, under the name extract B (LU105).
[0057] Finally, it has been noted, entirely surprisingly and
unexpectedly, that particularly advantageous results are obtained
when the active compound for inhibiting Langerhans cell migration
is a combination of at least one PKC-inhibiting compound, such as
those described above, with at least one MMP-inhibiting compound,
such as those described above.
[0058] It has thus been possible to note, as illustrated in the
examples below, that such a specific combination advantageously
makes it possible to potentiate, or even to provide a synergistic
effect of, the respective activities in order to thus obtain a
spectacular effect of inhibition of Langerhans cell migration,
which can be compared to complete extinction of the immune response
of reactive, sensitive and/or allergic skin, i.e. to the obtaining
of a normal skin reactivity.
[0059] In particular, the concentration of active compound for
inhibiting Langerhans cell migration is between approximately 0.001
and approximately 10% by weight, and more particularly between
approximately 0.01 and 3% by weight, relative to the total weight
of the pharmaceutical or cosmetic composition.
[0060] Preferably, the composition according to the invention is
characterized in that the PKC-inhibiting active compound is
sphingosine and/or isomers thereof, and said MMP-inhibiting
compound is a peptide extract of lupin, more preferably the extract
B (LU105).
[0061] The composition according to the present invention may also
comprise at least one pharmaceutically, in particular
dermatologically, or cosmetically acceptable excipient. Any
excipient suitable for the pharmaceutical forms known to those
skilled in the art, for the purpose of topical, oral, enteral or
parenteral administration, can be used.
[0062] In particular, the excipient may be suitable for obtaining a
composition in the form of an oily or aqueous solution, of a
water-in-oil emulsion or an oil-in-water emulsion, of a
microemulsion, of an oily or aqueous gel, of an anhydrous gel, of a
cream, of a lotion, of a spray, of a mask, of a milk, of a
dispersion of vesicles, of microcapsules or of micro-particles, or
else of gel capsules or of soft gelatin or plant capsules.
[0063] Preferably, an excipient suitable for external topical
administration is used.
[0064] Finally, the composition according to the present invention
may also comprise at least one pharmaceutically or cosmetically
acceptable adjuvant known to those skilled in the art, such as
thickeners, preserving agents, fragrances, dyes, chemical or
mineral screening agents, moisturizers, thermal spring waters,
etc.
[0065] A subject of the present invention is also the use of an
active compound chosen from the group consisting of protein kinase
C(PKC)-inhibiting compounds, in particular of those described
above, matrix metalloprotease (MMP)-inhibiting compounds, in
particular described above, and combinations thereof, for preparing
a composition intended to inhibit Langerhans cell migration.
[0066] The concentration of active compound used according to the
invention is between approximately 0.001 and approximately 10% by
weight, and more particularly between 0.01 and 3% by weight,
relative to the total weight of the pharmaceutical or cosmetic
composition.
[0067] The composition thus prepared may also comprise at least one
pharmaceutically, in particular dermatologically, or cosmetically
acceptable excipient, and also at least one adjuvant, as described
above.
[0068] In particular, the composition prepared by the use according
to the invention is intended for the treatment and prevention of
allergic reaction of the skin and of the mucous membranes (mouth,
lungs, bladder, rectum, vagina), insofar as it makes it possible to
reduce an allergic response in particular induced by LC
migration.
[0069] More particularly, the composition prepared by the use
according to the invention is intended for the treatment and
prevention of atopic eczema, insofar as it makes it possible to
reduce an immune response in particular induced by the migration of
LCs which have attached IgEs at the surface.
[0070] The composition prepared by the use according to the
invention is also intended for the treatment and prevention of
contact eczema, insofar as it makes it possible to reduce an immune
response in particular induced by capture of an antigen,
processing, and presentation of this antigen to T lymphocytes by
the LCs.
[0071] The composition prepared by the use according to the
invention is also intended for the treatment and prevention of
sensitive/reactive skin.
[0072] The composition prepared by the use according to the
invention is also intended for the treatment and prevention of
inflammatory dermatoses and/or irritant dermatitis.
[0073] It should therefore be noted that the composition described
above may advantageously be used as an additional component in a
pharmaceutical or cosmetic product or alternatively a fragrance, in
particular for external topical use, containing a main active
compound which is allergenic in nature. The allergic reaction
thereof will thus advantageously be accordingly decreased, or the
dose of main active compound which is allergenic in nature may thus
even advantageously be increased.
[0074] The composition prepared by the use according to the
invention is also intended for the treatment and prevention of
autoimmune diseases or inflammatory diseases such as psoriasis.
[0075] The composition prepared by the use according to the
invention is also intended for the prevention of
photoimmunosuppression.
[0076] Finally, the composition prepared by the use according to
the invention is also intended for the prevention of transplant
rejection.
[0077] The following examples are intended to illustrate the
present invention and can in no way be interpreted as being able to
limit the scope thereof. FIG. 1 is a histogram illustrating the
migratory indices of the LCs measured as described in example 1: 1:
control cells; 2: cells sensitized with the hapten DNSB; 3:
DNSB+LU105 (5 .mu.g/ml); 4: DNSB+D-sphingosine (2.5 .mu.M); 5:
DNSB+D-sphingosine (2.5 .mu.M)+LU105 (5 .mu.g/ml).
EXAMPLE 1
Study of the Activity of a Combination of a PKC Inhibitor with an
MMP Inhibitor on the Inhibition of LC Migration
[0078] 1) Materials and Methods
[0079] 1.1 Production of Suspension Enriched in LCs
[0080] Suspensions of epidermal cells were obtained by enzyme
treatment (0.05% trypsin, for 18 h at +4.degree. C.) of fragments
of normal human skin derived from plastic surgery. The suspensions
obtained contain, on average, 2 to 4% of LCs. The production of
suspensions containing, on average, 70% of LCs is based on the
principle of density gradient centrifugation (Lymphoprep.TM.) and
elimination of keratinocytes.
[0081] 1.2 Preparation of Media
[0082] The basic medium chosen for the entire study was RPMI 1640
(Gibco BRL, France).
[0083] D-sphingosine was diluted in ethanol so as to obtain, a
stock solution at 5.times.10.sup.-3 M. The D-sphingosine was used
at a concentration of 2.5 .mu.M, the dilution carried out in RPMI
containing 1% of bovine serum albumin (RPMI-BSA).
[0084] A peptide extract of lupin was used as MMP inhibitor. It is
the peptide extract LU105 from Laboratoires Pharmascience. LU105
was diluted in RPMI-1640 in order to obtain a stock solution at 250
mg/ml.
[0085] LU105 was used at a final concentration of 5 .mu.g/ml, the
dilution carried out in RPMI-1640. The cells were pre-incubated at
37.degree. C. for 60 min in the presence of LU105, before adding to
the medium the hapten DNSB as sensitizing agent.
[0086] 1.3 Sensitization of the LCs
[0087] As sensitizing agent, DNSB (Sigma Aldrich) was used, the
soluble form of DNCB (dinitrochlorobenzene), and was solubilized in
RPMI-BSA and used at a concentration of 50 .mu.M.
[0088] 1.4 Migration of the LCs
[0089] A two-compartment culture chamber system (Falcon, Becton
Dickinson, France) was used. The upper compartment is separated
from the lower compartment by a membrane with a porosity of 8
.mu.m, onto which are deposited 50 .mu.g/cm.sup.2 of Matrigel. The
membrane is then covered with proteins, forming a film equivalent
to a basal membrane (laminin, collagen IV, nidogen, entactin,
heparan sulfate proteoglycans). The cells taken up in the RPMI-BSA
medium alone or in the presence of the various products are placed
in the upper compartment. Normal human fibroblast culture
supernatant is added to the lower compartment. After incubation for
18 h at 37.degree. C., the number of living cells which have
crossed the Matrigel and are in the lower compartment is counted
under a microscope (the LCs are easily identifiable by their
dendritic shape). Each assay is carried out in triplicate.
[0090] 2) Results
[0091] 2.1 The results are given in table 3 below, and illustrated
by the histogram of FIG. 1.
TABLE-US-00003 TABLE 3 LC migration index 1 2 3 4 5 Migration index
1 1.61 1.38 1.15 0.88 Legend for table 3 and for the histogram of
FIG. 1 1: Control cells 2: Cells sensitized with the hapten DNSB 3:
DNSB + LU105 (5 .mu.g/ml) 4: DNSB + D-sphingosine (2.5 .mu.M) 5:
DNSB + LU105 (5 .mu.g/ml) + D-sphingosine (2.5 .mu.M)
[0092] 2.2 Migration of the LCs
[0093] The results represent the ratio between the number of cells
having migrated in the presence of DNSB.+-.D-sphingosine or LU105
or the combination D-sphingosine+LU105 and the number of cells
having migrated under the normal conditions (nonsensitized,
nontreated control cells). LCs freshly isolated from the epidermis
do not have a high migratory capacity. In expressing the results,
the migratory capacity of the control (nontreated and
nonsensitized) LCs is arbitrarily fixed at 1.
[0094] Treatment of the cells with the hapten DNSB significantly
stimulated LC migration (increase of 61%) compared to the normal
unstimulated cells (control cells).
[0095] D-sphingosine at concentrations of 2.5 .mu.M (table 3)
significantly inhibits the LC migration induced by DNSB.
[0096] In an experiment in which the cells were not brought into
contact with the hapten DNSB, but with LU105 alone (three
concentrations), no modification of the migration index, compared
to the control (nonsensitized and nontreated) cells, was observed.
This indicates that, in the absence of stimulation of the LCs,
LU105 has no effect on the migration. LU105 (5 .mu.g/ml)
significantly inhibits LC migration induced by DNSB.
[0097] The combination D-sphingosine+LU105 makes it possible to
completely inhibit the effect of DNSB on the LCs, the number of LCs
having migrated in the presence of DNSB+D-sphingosine+LU105 being
similar to that of the control (nonsensitized and nontreated)
cells. This combination therefore has a potentiating effect on the
intrinsic properties of the two products tested.
[0098] 3) Conclusions
[0099] In this study, we have demonstrated, using freshly isolated
LCs placed in a two-compartment culture chamber system (allowing
cell migration), that D-sphingosine (PKC inhibitor) potentiates the
inhibitory effect of LU105 (MMP inhibitor) on the migration of LCs
sensitized with the hapten DNSB, and vice versa. Specifically,
these two molecules used separately significantly inhibit LC
migration. When they are combined, the sensitized LCs have a
migratory capacity similar to that of nonactivated LCs. In other
words, according to this example, the combination of a PKC
inhibitor and an MMP inhibitor makes it possible to completely
extinguish the immune response of reactive/sensitive and allergic
skin, and therefore to return to a situation of normal skin.
* * * * *