U.S. patent application number 13/824627 was filed with the patent office on 2013-07-18 for avocado flesh and/or skin extract rich in polyphenols and cosmetic, dermatological and nutraceutical compositions comprising same.
This patent application is currently assigned to LABORATOIRES EXPANSCIENCE. The applicant listed for this patent is Caroline Baudouin, Sebastien Garnier, Sophie Leclere-Bienfait, Philippe Msika, Alex Saunois. Invention is credited to Caroline Baudouin, Sebastien Garnier, Sophie Leclere-Bienfait, Philippe Msika, Alex Saunois.
Application Number | 20130183255 13/824627 |
Document ID | / |
Family ID | 44246374 |
Filed Date | 2013-07-18 |
United States Patent
Application |
20130183255 |
Kind Code |
A1 |
Saunois; Alex ; et
al. |
July 18, 2013 |
AVOCADO FLESH AND/OR SKIN EXTRACT RICH IN POLYPHENOLS AND COSMETIC,
DERMATOLOGICAL AND NUTRACEUTICAL COMPOSITIONS COMPRISING SAME
Abstract
The invention relates to an avocado extract rich in polyphenols,
containing at least 10% by weight of polyphenols, expressed in
gallic acid equivalent relative to the dry extract obtained,
wherein said polyphenols contain procyanidins, cafeic acid and
cafeic acid derivatives, typically in a proportion of at least 70%
by weight, expressed in gallic acid equivalent relative to the
total polyphenol content by weight. The invention also relates to a
composition comprising an extract according to the invention as
active agent and a suitable excipient. The invention also relates
to such a composition or such an extract for use thereof in
preventing or treating disorders or pathological conditions of the
skin, the mucous membranes or the superficial body growths.
Finally, the invention relates to a cosmetic care process for the
skin, the superficial body growths or the mucous membranes, with a
view to improving the condition thereof or the appearance thereof,
which consists in administering such a composition or such an
extract.
Inventors: |
Saunois; Alex;
(Nogent-Le-Roi, FR) ; Baudouin; Caroline;
(Rambouillet, FR) ; Leclere-Bienfait; Sophie;
(Dreux, FR) ; Garnier; Sebastien; (Pierres,
FR) ; Msika; Philippe; (Versailler, FR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Saunois; Alex
Baudouin; Caroline
Leclere-Bienfait; Sophie
Garnier; Sebastien
Msika; Philippe |
Nogent-Le-Roi
Rambouillet
Dreux
Pierres
Versailler |
|
FR
FR
FR
FR
FR |
|
|
Assignee: |
LABORATOIRES EXPANSCIENCE
Courbevoie
FR
|
Family ID: |
44246374 |
Appl. No.: |
13/824627 |
Filed: |
December 22, 2011 |
PCT Filed: |
December 22, 2011 |
PCT NO: |
PCT/EP2011/073832 |
371 Date: |
March 18, 2013 |
Current U.S.
Class: |
424/59 ; 514/188;
514/456 |
Current CPC
Class: |
A61Q 19/005 20130101;
A61K 9/0014 20130101; A23L 33/105 20160801; A61K 8/9794 20170801;
A61K 2800/92 20130101; A61Q 17/04 20130101; A61K 31/353 20130101;
A61Q 19/00 20130101; A23L 19/07 20160801; A61P 17/00 20180101; A23L
19/09 20160801; A61K 8/9789 20170801; A61K 36/54 20130101; A61Q
19/08 20130101; A61Q 17/00 20130101; A23V 2002/00 20130101; A61K
45/06 20130101; A61Q 5/12 20130101; A61K 8/498 20130101; A61P 17/10
20180101; A61Q 19/02 20130101; A23V 2002/00 20130101; A23V 2200/318
20130101; A23V 2250/21 20130101 |
Class at
Publication: |
424/59 ; 514/456;
514/188 |
International
Class: |
A61K 31/353 20060101
A61K031/353; A61Q 5/12 20060101 A61Q005/12; A61K 8/49 20060101
A61K008/49; A61Q 19/00 20060101 A61Q019/00; A61Q 17/04 20060101
A61Q017/04; A61K 45/06 20060101 A61K045/06 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 22, 2010 |
FR |
1061055 |
Claims
1. A polyphenol-rich extract of avocado fruit, containing at least
10% by weight of polyphenols and advantageously between 10 and 30%
by weight of polyphenols, expressed in gallic acid equivalents in
relation to the dry extract obtained, said polyphenols
advantageously containing procyanidins, caffeic acid and
derivatives of caffeic acid, typically in a proportion of at least
70% by weight, expressed in gallic acid equivalents in relation to
the total polyphenols content by weight.
2. The extract of claim 1, characterized in that the proportions of
procyanidins, advantageously selected from the group comprised of
B-type procyanidin dimers, A- and B-type procyanidin trimers and A-
and B-type procyanidin tetramers, of caffeic acid and of
derivatives of caffeic acid in said polyphenols are at least 80% by
weight, expressed in gallic acid equivalents in relation to the
total polyphenols content by weight.
3. The extract of claim 1 or claim 2, characterized in that the
polyphenols contained in this extract contain at least 30% of
procyanidins, expressed in gallic acid equivalents in relation to
the total polyphenols content by weight.
4. The extract of any one of the preceding claims, characterized in
that the polyphenols contained in this extract contain caffeic acid
and derivatives thereof in a proportion of at least 30% by weight,
expressed in gallic acid equivalents in relation to the total
polyphenols content by weight.
5. The extract of any one of the preceding claims, characterized in
that it further includes at least 10%, advantageously from 10 to
60% avocado sugars, said sugars containing at least
D-mannoheptulose and/or perseitol, with percentages being expressed
by weight in relation to the weight of the dry extract.
6. The extract of any one of the preceding claims, characterized in
that it is obtained by extraction of the avocado fruit,
advantageously by extraction of avocado fruits that first have been
dried, under gentle conditions, and then delipidated.
7. The extract of any one of claims 1 to 6, characterized in that
it is obtained by solid-liquid extraction of a portion of the
avocado fruit in an aqueous and/or alcoholic and/or glycolic and/or
glycerolic solvent.
8. The extract of claim 7, characterized in that the solvent is
selected from the group composed of water, ethanol, glycerol or a
glycol such as propanediol, or mixtures thereof, advantageously
binary mixtures, in proportions between 0 and 100% of water in
relation to other solvents.
9. A composition including as active agent an extract as defined in
any one of claims 1 to 8 and a suitable carrier, characterized in
that it is a cosmetic, pharmaceutical, dermatological or
nutraceutical composition.
10. The composition of claim 9, further including another active
agent, in particular selected from the group composed of
emollients, moisturizing active agents, keratoregulators,
keratolytics, healing agents and/or agents that restructure the
cutaneous barrier, PPAR, RXR or LXR agonists, sebum-regulating
agents, anti-irritation and/or anti-inflammatory and/or soothing
agents, antioxidant agents, anti-aging agents, depigmenting or
hypopigmenting agents, pigmenting agents, lipolytic or lipogenesis
inhibitor agents or anti-cellulitis or slimming agents, organic or
mineral sun screens and filters (pigmentary or ultrafine),
antifungal compounds, preservatives, antibacterial agents,
prebiotics and probiotics, antibiotics, and immunomodulators.
11. The composition of claim 9 or claim 10, characterized in that
the other active agent is selected from: healing agents and/or
agents that restructure the cutaneous barrier, preferably
panthenol, arabinogalactan, zinc oxide, ceramides, cholesterol,
squalane and phospholipids, sebum-regulating agents, preferably
selected from 5-alpha reductase inhibitors and zinc derivatives,
anti-inflammatory and/or anti-irritation and/or soothing agents,
preferably arabinogalactan, hypopigmenting or depigmenting agents,
preferably N-undecylenoyl-L-phenylalanine, organic or mineral sun
screens and filters, preferably UVB and/or UVA sun screens and
filters, and preservatives preferably selected from capryloyl
glycine, glyceryl caprylate and hexanediol.
12. The composition of any one of claims 9 to 11, further including
at least one other active agent selected from the group composed
of: plant oils, preferably soy oil, rapeseed oil, avocado oil,
lupin oil, and advantageously sweet white lupin oil, or a mixture
of these oils, oleodistillates or concentrates of plant or animal
oil, preferably of sunflower, avocado, rapeseed, corn and palm and
advantageously concentrated in unsaponifiables, unsaponifiables of
plants or plant oil, preferably avocado unsaponifiables, soy
unsaponifiables or mixtures thereof, advantageously of avocado
furans, and in particular, a mixture of avocado furanic
unsaponifiables and soy unsaponifiables in a respective ratio of
approximately 1/3-2/3, sterolic unsaponifiables, phytosterols,
esters of sterols and vitamin derivatives, peptides or complexes of
plant amino acids, preferably avocado peptides, lupin peptides,
quinoa peptides, maca peptides, fermented or non-fermented soy
peptides, rice peptides, plant sugars, preferably avocado sugars,
butyl avocadate, polyphenol-rich extracts, preferably extracts of
the above-ground parts of Gynandropsis gynandra and extracts of
maca leaves, lupeol, total lupin extract, extract of Acacia
macrostachya seeds, extract of maca leaves, an extract of
Schisandra sphenanthera and an extract of Vigna unguiculata seeds,
oxazolines, preferably
2-undecyl-4-hydroxymethyl-4-methyl-1,3-oxazoline,
2-undecyl-4,4-dimethyl-1,3-oxazoline,
(E)-4,4-dimethyl-2-heptadec-8-enyl-1,3-oxazoline,
4-hydroxymethyl-4-methyl-2-heptadecyl-1,3-oxazoline,
(E)-4-hydroxymethyl-4-methyl-2-heptadec-8-enyl-1,3-oxazoline,
2-undecyl-4-ethyl-4-hydroxymethyl-1,3-oxazoline and
2-undecyl-4,4-dimethyl-1,3-oxazoline, and mixtures thereof.
13. The composition of any one of claims 9 to 12, characterized in
that it is formulated to be administered by topical or oral
route.
14. The composition of any one of claims 9 to 13 or extract as
defined in any one of claims 1 to 8 to be used in the prevention
and/or treatment of disorders or pathologies of the skin and/or
mucous membranes and/or appendages, advantageously allergic,
inflammatory or irritative reactions or pathologies or disorders of
the barrier or homeostasis of the skin, or vascular disorders or as
depigmenting and healing agent.
15. A cosmetic care method for the skin and/or appendages and/or
mucous membranes, advantageously for sensitive skin, with a view to
improving the condition and/or appearance thereof, in particular in
order to promote the hydration thereof, or to improve the firmness,
elasticity or tonicity of the skin and to prevent the accumulation
of adipose tissue and skin with cellulitis, or to reduce
pigmentation spots, or to prevent and/or treat aging, comprising
the administration of a cosmetic composition according to any one
of claims 9 to 13 or an extract as defined in any one of claims 1
to 8.
Description
[0001] The invention relates to an extract of avocado fruit,
notably avocado flesh and/or skin, rich in polyphenols, and to
cosmetic, pharmaceutical, dermatological and nutraceutical
compositions including such an extract and optionally a suitable
carrier.
[0002] The invention also has as an object a method for extracting
an avocado extract rich in polyphenols, as well as the extract
likely to be obtained by said method.
[0003] The invention also relates to such a composition or such an
extract for use thereof in the prevention or treatment of disorders
or pathologies of the skin, mucous membranes or appendages.
Finally, the invention relates to a cosmetic care method for the
skin, appendages or mucous membranes, in order to improve the
condition or appearance thereof, consisting of administering such a
composition or such an extract.
[0004] The avocado tree (Persea americana or Persea gratissima)
belongs to the family Lauraceae.
[0005] There is a great diversity of avocado varieties. Among the
most widespread, mention may be made of the Hass, Fuerte, Ettinger,
Bacon, Nabal, Ananheim, Lula, Reed, Zutano, Quenn, Criola Selva,
Mexicana Canta, Tegion Dschang, Hall, Booth, Peterson and Collinson
Redn varieties. Particularly, the Hass, Fuerte, Ettinger and Bacon
varieties will be selected, and more advantageously the Hass and
Fuerte varieties. The weight of the fruits may vary from 50 g to
more than 1 kg. The fruit consists of 10 to 20% seed, 7 to 15%
integument and 65 to 80% flesh. The skin is of variable thickness,
texture and color, smooth to rough, in general green but may become
brown, purplish to black.
[0006] The flesh is of variable flavor and color and is on average
composed of:
TABLE-US-00001 TABLE 1 Composition of avocado flesh; *Wei Wang,
Food Chemistry, 122: 1193-1198 (2010) Water 70-85% Proteins
1.5-4.5% Lipids 12-23% Sugars 1.5-5% Fibers 1.1-1.6% Polyphenols
Between 0.06 and 0.5% in fresh flesh Highly variable =
(varieties)
[0007] The flesh of the avocado is widely used by South American
population to give the hair shine. Topical applications of avocado
flesh on the skin are also very common for its antioxidant and
nutritive activity, related to the nature of the lipids of which it
is composed (fatty acids and unsaponifiable fraction-vitamin
E).
[0008] The fruits, in particular their flesh, are commonly used in
home remedies in the tropics. The fruits are very rich in vitamins
A, B, C, D, E and K and are recommended for diabetics. The flesh is
also acaricidal (and vermifugal), aphrodisiac and tonic. It is used
in South America in poultices to treat boils and whitlow.
[0009] It has been shown that a high consumption of avocados has an
effect on blood cholesterol levels. In particular, after 7 days of
an avocado-rich diet, patients with hypercholesterolemia saw their
total cholesterol levels decrease by 17%. These subjects also
showed a 22% decrease in (bad) LDL cholesterol levels and
triglycerides, and an 11% increase in (good) HDL cholesterol. Its
monounsaturated fatty acids are excellent for cardiovascular health
and good blood cholesterol (HDL). Moreover, the avocado is a very
good source of fibers (soluble and insoluble) and antioxidants.
[0010] The term "polyphenols" refers to a vast family of molecules,
widespread in the plant world, characterized by the presence of one
or more phenolic nuclei, notably comprising the following
subfamilies: [0011] phenolic acids (cinnamic and benzoic); [0012]
flavonoids, near-universal plant pigments, responsible for the
colors of flowers, fruits and sometimes leaves. Anthocyans are
members of this family (blue to violet in color, passing through
red); [0013] quinones and anthraquinones and certain derived
condensed forms; and [0014] more complex and polymerized forms:
hydrolysable tannins derived from gallic acid and ellagic acid;
condensed tannins derived from catechic acid, OPC or
proanthocyanidols.
[0015] These molecules are known for their numerous biological
activities (antioxidant, anti-inflammatory, antibacterial) and are
used in the fields of pharmacy, cosmetics and nutrition/dietary
supplements as a function of certain specific structures.
[0016] Polyphenols are widely distributed in numerous families of
the higher plant kingdom because they are often essential to their
survival, namely protection against various external attacks (UV
rays, drought, microorganisms, etc.).
[0017] Among the most widespread and used polyphenols, mention may
be made of polyphenols from tea (Camellia sinensis), grape (Vitis
vinifera), cocoa (Theobroma cacao). The application FR 2 928 263
describes compositions including a combination of plant polyphenols
(apple polyphenols are described in particular) and plant sugars.
The polyphenols sought are polymeric polyphenols, and in particular
procyanidins and prodelphinidins. The avocado is not cited as a
source of sugars.
[0018] The inventors have discovered that polyphenol-rich extracts
of avocado (Persea gratissima or americana) have cosmetic,
dermatological, pharmaceutical and nutraceutical properties, in
particular cosmetic and dermatological properties, never described
to date. In particular, it is the first time that polyphenol-rich
extracts of avocado have been used as such, for their specific
properties.
[0019] The objects of the invention are in particular: [0020] a
polyphenol-rich extract of avocado fruit, containing at least 10%
by weight of polyphenols, expressed in gallic acid equivalents in
relation to the dry extract obtained, said polyphenols
advantageously being procyanidins, caffeic acid and derivatives of
caffeic acid, typically in a proportion of at least 70% by weight
expressed in gallic acid equivalents in relation to the total
polyphenols content by weight. In particular, the extract contains
10 to 30% by weight of polyphenols expressed in gallic acid
equivalents in relation to the weight of the dry extract. The
proportions of procyanidins, caffeic acid and derivatives of
caffeic acid in said polyphenols are advantageously at least 80% by
weight expressed in gallic acid equivalents in relation to the
total polyphenols content by weight. [0021] The procyanidins are
advantageously selected from the group composed of B-type
procyanidin dimers, A- and B-type procyanidin trimers and A- and
B-type procyanidin tetramers. [0022] The polyphenols contained in
this extract advantageously contain at least 30% by weight of
procyanidins, expressed in gallic acid equivalents in relation to
the total polyphenols content by weight. [0023] The polyphenols
contained in this extract advantageously contain caffeic acid and
derivatives thereof, typically in a proportion of at least 30% by
weight, expressed in gallic acid equivalents in relation to the
total polyphenols content by weight. [0024] The extract
advantageously further includes at least 10%, advantageously from
10 to 60% of avocado sugars, said sugars containing at least
D-mannoheptulose and/or perseitol, with percentages being expressed
by weight in relation to the total weight of the dry extract.
[0025] The extract is advantageously obtained by extraction of the
avocado fruit, advantageously by extraction of avocado fruits that
first have been dried, under gentle conditions, and then
delipidated. [0026] In one advantageous variant, it is obtained by
solid-liquid extraction of part of the avocado fruit in an aqueous
and/or alcoholic and/or glycolic and/or glycerolic solvent. The
solvent is advantageously selected from the group composed of
water, ethanol, glycerol or a glycol such as propanediol, and
mixtures thereof, such as binary mixtures, in proportions between 0
and 100% of water in relation to other solvents. [0027] a
composition including as active agent an extract of the invention
and, advantageously a suitable carrier. In particular, the
composition is a cosmetic, pharmaceutical, dermatological or
nutraceutical composition. [0028] The composition advantageously
further includes another active agent, in particular selected from
the group composed of emollients, moisturizing active agents,
keratoregulators, keratolytics, healing agents and/or agents that
restructure the cutaneous barrier, PPAR, RXR or LXR agonists,
sebum-regulating agents, anti-irritation and/or anti-inflammatory
and/or soothing agents, antioxidant agents, anti-aging agents,
depigmenting or hypopigmenting agents, pigmenting agents, lipolytic
or lipogenesis inhibitor agents or anti-cellulitis or slimming
agents, organic or mineral sun screens and filters (pigmentary or
ultrafine), antifungal compounds, preservatives, antibacterial
agents, prebiotics and probiotics, antibiotics, and
immunomodulators. [0029] The composition advantageously includes as
another active agent an agent useful in the treatment of acne, in
particular selected from the group comprised of 5-alpha reductase
inhibitors, zinc (and gluconate, salicylate salts thereof, and
pyroglutamic acid), spironolactone, linoleic acid, antibiotics,
benzoyl peroxide, azelaic acid, vitamin PP, vitamin B3, cyclins,
extracts of pumpkin seeds, squash seeds oil and palmetto. [0030] an
extract or a composition of the invention to be used as a
pharmaceutical, dermatological or cosmetic composition or as a
functional food; in particular to be used in the prevention and/or
treatment of disorders or pathologies of the skin and/or mucous
membranes and/or appendages, more particularly to be used in the
prevention and/or treatment of allergic, inflammatory, irritative
reactions or pathologies, or disorders of the barrier or
homeostasis of the skin, or vascular disorders, or as a
depigmenting or healing agent. [0031] a cosmetic care method for
the skin and/or appendages and/or mucous membranes, with a view to
improving the condition and/or appearance thereof, consisting of
administering a composition or an extract of the invention.
[0032] The expression "polyphenol-rich extract of avocado" refers
to an extract obtained by methods making it possible to concentrate
the polyphenols potentially present in the avocado in such a way
that this extract contains at least 10% by weight of polyphenols,
expressed in gallic acid equivalents in relation to the weight of
the dry extract. According to one advantageous variant of the
invention, the extract contains 10 to 30% by weight of polyphenols,
more advantageously 10 to 20% by weight of polyphenols, expressed
in gallic acid equivalents in relation to the dry extract
obtained.
[0033] The proportion of dry extract in the extract of the
invention, expressed by weight in relation to the total weight of
the extract, varies from 0.01 to 90%, advantageously from 0.5 to
50%, more advantageously from 0.5 to 15%, even more advantageously
from 0.5 to 5%.
[0034] The polyphenols contained in this extract contain
procyanidins, caffeic acid and derivatives of caffeic acid, in
particular esterified derivatives, in a proportion of at least 70%,
advantageously at least 80%, more advantageously at least 90%, even
more advantageously at least 95%, by weight expressed in gallic
acid equivalents in relation to the total polyphenols content by
weight.
[0035] Procyanidins are oligomers of catechins/epicatechins.
Catechin and epicatechin are, respectively, the (+) and (-) optical
isomers of the molecule of the following formula (I):
##STR00001##
[0036] The procyanidins are advantageously B-type procyanidin
dimers. Procyanidins of the B-1, B-2, B-3 and B-4 types are dimers
composed of two units (+)-catechin and (-)-epicatechin, linked in
C.sub.4-C.sub.8. Procyanidins of the B-5, B-6, B-7 and B-8 types
are dimers composed of two units (+)-catechin and (-)-epicatechin,
linked in C.sub.4-C.sub.6.
[0037] The procyanidins are more advantageously A and B type
procyanidin trimers.
[0038] The procyanidins are more advantageously A and B type
procyanidin tetramers.
[0039] The procyanidins are more advantageously oligomeric or
polymeric procyanidins (number of units of catechin or epicatechin
>4).
[0040] The polyphenols contained in this extract advantageously
contain at least 30%, more advantageously at least 50% by weight of
procyanidins, expressed in gallic acid equivalents in relation to
the total polyphenols content by weight.
[0041] The polyphenols contained in this extract advantageously
also contain caffeic acid and derivatives thereof in a proportion
of at least 20%, more advantageously at least 30% by weight,
expressed in gallic acid equivalents in relation to the total
polyphenols content by weight.
[0042] Caffeic acid has the following formula (II):
##STR00002##
[0043] Caffeic acid and its derivative, phenethyl caffeate ester
(ester of caffeic acid and 2-phenylethanol), are present in many
plants, and in particular in a large quantity in coffee, from which
it gets its name. Caffeic acid may be present in esterified form,
for example with quinic acid with which it forms chlorogenic acid.
Among other esterified forms, mention may be made of caffeoyl
tartaric (or caftaric) acid formed with tartaric acid, caffeoyl
shikimic acid (formed with shikimic acid), caffeoyl malic acid
(formed with malic acid), or esterified with glucose (caffeoyl
glucose) or in amide form with putrescine (caffeoyl
putrescine).
[0044] Other polyphenols may also be present in the extract of the
invention but they are, however, a minority. Their content by
weight, expressed in gallic acid equivalents in relation to the
total polyphenols content by weight, is advantageously less than
30%, more advantageously less than 20%, even more advantageously
less than 10%, even more advantageously less than 5%, even more
advantageously less than 2%.
[0045] According to one advantageous variant of the invention, the
extract contains at least 10%, advantageously from 10 to 60%, more
advantageously from to 30% of avocado sugars, typically said sugars
containing at least D-mannoheptulose and/or perseitol (percentages
are expressed by weight in relation to the weight of the dry
extract).
[0046] In the context of the present invention, the terms
"D-mannoheptulose" and "perseitol" also cover the chemical
derivatives thereof.
[0047] The avocado sugars are mainly D-mannoheptulose, perseitol,
sucrose, glucose and fructose. Traces of other sugars may be found,
but their content is less than 5% by weight, advantageously less
than 2% by weight, in relation to the total weight of avocado
sugars.
[0048] In the extract of the invention, the total proportion of
D-mannoheptulose and/or perseitol in the avocado sugars is
advantageously at least 50% by weight in relation to the total
sugars content by weight. More advantageously, the total proportion
of D-mannoheptulose and/or perseitol in the avocado sugars is from
50 to 98% by weight, more advantageously 55 to 95% by weight, even
more advantageously 60 to 90% by weight, in relation to the total
sugars content by weight.
[0049] The weight ratio of D-mannoheptulose to perseitol varies
advantageously from 1:10 to 10:1.
[0050] The extract of the invention advantageously contains 0 to
50% by weight, more advantageously 0 to 20% by weight, even more
advantageously 0 to 10% by weight of avocado lipids, with
percentages being expressed by weight in relation to the total
weight of the dry extract.
[0051] The extract of the invention advantageously contains 0 to
60% by weight, more advantageously 1 to 30% by weight, even more
advantageously 5 to 15% by weigh of avocado proteins, with
percentages being expressed by weight in relation to the total
weight of the dry extract (Bradford assay).
[0052] The extract of the invention is obtained by extraction of a
portion of the avocado fruit, preferentially the flesh and/or skin.
More particularly, the residual oil cake from the pressing of dried
avocado flesh is used. Preferentially, said flesh has been dried
beforehand before the lipid extraction.
[0053] It is particularly advantageous to use avocado fruits (flesh
and/or skin) that first have been dried, under gentle conditions,
and then delipidated. Indeed, it has been noted that under such
conditions the optimal preservation of polyphenols can be ensured
by inactivation of polyphenol oxidase during drying (even extracts
containing no polyphenol oxidase can be obtained).
[0054] Advantageously, this extract is obtained by solid-liquid
extraction of a portion of the avocado fruit in an aqueous and/or
alcoholic and/or glycolic and/or glycerolic solvent. The portions
of the fruit preferentially selected are dried and/or defatted
flesh (oil cake) and/or the skin, more preferentially flesh dried
under gentle conditions and then defatted.
[0055] The solvent is advantageously selected from the group
composed of water, ethanol, glycerol or a glycol such as
propanediol, and mixtures thereof, such as binary mixtures, in
proportions between 0 and 100% of water in relation to other
solvents.
[0056] Mainly, binary mixtures of solvents composed of water and a
solvent selected from ethanol, glycerol or propanediol are
used.
[0057] More particularly, between 0.1 and 50% by weight (expressed
in relation to the total weight of avocado+solvent) of the dried
portion of avocado is introduced into the extraction solvent, and
preferentially 10% by weight.
[0058] In the presence of ethanol, preferentially a proportion
between 30 and 70% of ethanol in water, and advantageously between
50 and 60% of ethanol in water, will be chosen (percentages are
expressed by weight of ethanol in relation to the total weight of
water+ethanol).
[0059] In the presence of glycerol, preferentially a concentration
between 40 and 90% of glycerol, and preferentially between 60 and
80%, will be chosen (percentages are expressed by weight of
glycerol in relation to the total weight of water+glycerol).
[0060] In the presence of glycol, and more particularly of
propanediol, preferentially a proportion between 40 and 80%, and
advantageously between 50 and 60% of propanediol in relation to
water will be chosen (percentages are expressed by weight of
propanediol in relation to the total weight of
water+propanediol).
[0061] The extraction temperature is advantageously between 4 and
100.degree. C., and preferentially between 10 and 60.degree. C.,
and more particularly between 15 and 30.degree. C.
[0062] The extraction period advantageously varies from minutes to
4 hours, more particularly from 30 minutes to 2 hours.
[0063] These various extractions may be followed by purification
steps of ultrafiltration (10 kDa cut-off, for example), or
diafiltration and/or nanofiltration (200 Da cut-off, for example),
making it possible to concentrate the polyphenols at the expense of
sugars (including the specific C.sub.7 avocado sugars,
mannoheptulose and perseitol).
[0064] The extract obtained following these purification steps
advantageously includes 10 to 30% by weight of polyphenols, even
more advantageously 10 to 20% by weight of polyphenols, expressed
in gallic acid equivalents in relation to the weight of the dry
extract.
[0065] The sugars content in the extract obtained following these
purification steps is advantageously between 5 and 20% by weight,
in relation to the weight of the dry extract.
[0066] The sugars present in this extract advantageously include 20
to 60% of D-mannoheptulose and/or perseitol, more advantageously 30
to 60% of D-mannoheptulose and/or perseitol, expressed by weight in
relation to the total weight of the sugars.
[0067] The extract obtained may be provided in liquid form, but
also may be dried according to methods known to persons skilled in
the art, such as atomization or freeze-drying, for example, with or
without a support such as maltodextrin.
[0068] The invention also has as an object a method for preparing a
polyphenol-rich avocado extract comprising the following sequential
steps: [0069] (a) liquid-phase dispersion in a suitable solvent of
avocado fruit (advantageously flesh and/or skin), and
advantageously dried defatted flesh (oil cake); [0070] (b)
subjection of the mixture obtained following step (a) to extraction
under stirring in an aqueous and/or alcoholic and/or glycolic
and/or glycerolic solvent, advantageously a mixture of water and a
solvent selected from the group composed of ethanol, glycerol, a
glycol (advantageously propanediol) and mixtures thereof,
advantageously in a proportion of 60% of these solvents in water;
[0071] (c) centrifugation of the extract obtained following step
(b) and then filtration, or filtration directly; [0072] (d) if the
need arises, subjection of the extract obtained following step (c)
to an ultrafiltration and/or diafiltration and/or nanofiltration
step; [0073] (e) following step (c) or (d), recovery of the
polyphenol-rich extract; and [0074] (f) optional drying of the
extract obtained following step (e) on a support or not.
[0075] The extract is advantageously used as an active agent in a
cosmetic, pharmaceutical, dermatological or nutraceutical
composition, which may include one or more suitable carriers. The
composition may further include at least one other active compound
in addition to the polyphenol-rich avocado extract. This other
compound may be selected from all the compounds and functional
equivalents thereof set forth below.
[0076] This other compound may be selected in particular from
active agents classically used in dermatology, pharmaceuticals,
cosmetics or nutraceuticals and known to persons skilled in the art
such as emollients, moisturizing active agents, keratoregulators,
keratolytics, healing agents and/or agents that restructure the
cutaneous barrier, PPAR, RXR or LXR agonists, sebum-regulating
agents, anti-irritation and/or anti-inflammatory and/or soothing
agents, antioxidant agents, anti-aging agents, depigmenting or
hypopigmenting agents, pigmenting agents, lipolytic or lipogenesis
inhibitor agents or anti-cellulitis or slimming agents, organic or
mineral sun screens and filters (pigmentary or ultrafine),
antifungal compounds, preservatives, antibacterial agents,
prebiotics and probiotics, antibiotics, and immunomodulators.
[0077] More particularly, the healing agents and/or agents that
restructure the cutaneous barrier that may be used in combination
are advantageously panthenol (vitamin B5), arabinogalactan, zinc
oxide, ceramides, cholesterol, squalane and phospholipids.
[0078] The sebum-regulating agents that may be used in combination
are advantageously selected from the group composed of 5-alpha
reductase inhibitors such as Zinc and zinc derivatives (gluconate,
salicylate salts thereof and pyroglutamic acid).
[0079] The anti-inflammatory and/or anti-irritation and/or soothing
agent may be arabinogalactan.
[0080] Among hypopigmenting or depigmenting agents, particular
mention may be made of N-undecylenoyl-L-phenylalanine
(Sepiwhite.RTM.)
[0081] The sun protection active agents that may be used in
combination are advantageously UVB and/or UVA filters or sun
screens, such as the mineral and/or organic screens or filters
known to persons skilled in the art, who will adapt their choice
and their concentrations according to the degree of protection
sought.
[0082] The preservatives that may be used in combination are, for
example, those generally used in cosmetics or nutraceuticals,
molecules with antibacterial activity (pseudo-preservatives) such
as caprylic derivatives like, for example, capryloyl glycine and
glyceryl caprylate; hexanediol, sodium levulinate, and copper and
zinc derivatives (gluconate and PCA).
[0083] Among the recommended active agents in combination with the
extract of the invention, mention may be made of plant extracts, in
particular: [0084] plant oils such as soy oil and/or rapeseed oil,
avocado oil (WO2004/012496, WO2004/012752, WO2004/016106,
WO2007/057439), lupin oil, advantageously sweet white lupin oil (WO
98/47479), or a mixture of these oils; [0085] oleodistillates or
concentrates of animal or plant oil, notably sunflower, more
advantageously linoleic sunflower concentrates, such as the
sunflower oil concentrated in unsaponifiables (Soline.RTM.) (see
the international application WO 01/21150) marketed by Laboratoires
Expanscience, oils concentrated in unsaponifiables as the avocado
oil, rapeseed oil, corn oil or palm oil, useful notably for their
moisturizing and/or emollient, healing and/or cutaneous barrier
restructure, anti-inflammatory and/or anti-irritation and/or
soothing activity; [0086] unsaponifiables of plants or of plant
oil, advantageously of avocado furans (Avocadofurane.RTM.), able to
be obtained by the method described in the international
application WO 01/21605, unsaponifiables of avocado and/or soy,
more particularly a mixture of furanic unsaponifiables of avocado
and unsaponifiables of soy, advantageously in a respective ratio of
about 1/3-2/3 (such as Piascledine.RTM.), unsaponifiables of soy
(such as obtained according to the method described in the
international application WO 01/51596), sterolic unsaponifiables
(typically unsaponifiables whose proportion of sterols,
methylsterols and triterpene alcohols is between 20 and 95% by
weight, preferably 45-65% by weight, in relation to the total
weight of the unsaponifiable), phytosterols, esters of sterols and
vitamin derivatives, notably useful for their healing and/or
restructure of the cutaneous barrier, anti-aging or
anti-inflammatory activity; [0087] peptides or complexes of plant
amino acids, in particular of avocado peptides (such as those
described in the international application WO2005/105123), lupin
peptides (such as those obtained according to the method described
in the application WO2005/102259), quinoa peptides (such as those
described in the international application WO2008/080974), maca
peptides (such as those described in the international application
WO2004/112742), fermented or non-fermented soy peptides, rice
peptides (such as those described in the international application
WO2008/009709), useful notably for their moisturizing and/or
emollient activity (avocado), keratoregulating activity (lupin,
quinoa), healing and/or cutaneous barrier restructuring activity
(maca, quinoa, soy), anti-inflammatory and/or anti-irritation
and/or soothing activity (lupin, quinoa), antioxidant activity
(avocado), anti-aging activity (lupin, maca) and pigmenting
activity (rice); [0088] plant sugars, in particular avocado sugars
(such as those described in the application WO2005/115421), useful
notably for their keratoregulator, healing and/or cutaneous barrier
restructure, anti-inflammatory and/or anti-irritation and/or
soothing property; [0089] butyl avocadate (5 alpha Avocute),
5-alpha reductase inhibitor (see WO 01/52837 and WO 02/06205)
typically regulator of the seborrheic secretions found increased in
acne and in dandruff; [0090] polyphenol-rich extracts, and more
particularly extracts of the above-ground parts of Gynandropsis
gynandra (FR 1 061 051) and extracts of maca leaves (FR 1 061 047);
[0091] lupeol (FR 2 822 821, FR 2 857 596) useful notably to
promote healing; [0092] total extract of lupin (such as those
described in the international application WO2005/102259),
particularly suitable for the treatment of irritations; [0093]
extract of Acacia macrostachya seeds (FR 0958525), extract of maca
leaves (FR 1 061 047), extract of Schisandra sphenanthera seeds (FR
0955343 and FR 0955344) and Vigna unguiculata seeds (FR
0958529).
[0094] Among the recommended active agents in combination with the
extract of the invention, mention may be made of oxazolines, in
particular those selected from the group comprised of
2-undecyl-4-hydroxymethyl-4-methyl-1,3-oxazoline,
2-undecyl-4,4-dimethyl-1,3-oxazoline,
(E)-4,4-dimethyl-2-heptadec-8-enyl-1,3-oxazoline,
4-hydroxymethyl-4-methyl-2-heptadecyl-1,3-oxazoline,
(E)-4-hydroxymethyl-4-methyl-2-heptadec-8-enyl-1,3-oxazoline,
2-undecyl-4-ethyl-4-hydroxymethyl-1,3-oxazoline, preferably
2-undecyl-4,4-dimethyl-1,3-oxazoline (OX-100 or Cycloceramide.RTM.;
WO2004050052, WO2004050079 and WO2004112741). They are particularly
useful for their anti-inflammatory and/or anti-irritation and/or
soothing, antioxidant, depigmenting, immunomodulatory activity.
[0095] Among the active agents recommended in combination with the
extract of the invention, mention may be made of 5-alpha reductase
inhibitors such as butyl avocadate (5 alpha Avocuta.RTM.)
[0096] All of these combinations include at least one other active
compound, in addition to the polyphenol-rich avocado extract, and
may include two, three, four or more active compounds as described
above.
[0097] The composition of the invention may be formulated in the
form of various preparations suitable for topical administration,
for oral, rectal, vaginal, nasal, auricular or bronchial
administration, as well as for parenteral administration.
[0098] According to a first variant, the various preparations are
suitable for topical administration and notably include creams,
emulsions, milks, pomades, lotions, oils, aqueous or
water-alcoholic or glycolic solutions, powders, patches, sprays,
shampoos, varnishes or any other product for external
application.
[0099] According to a second variant, the various preparations are
suitable for oral administration, with the polyphenol-rich avocado
extract incorporated either in a dietary supplement or in a
nutraceutical composition. In the context of the present invention,
the dietary supplement may be provided in the form of the
polyphenol-rich avocado extract as such or in the form of hard or
soft plant- or gelatin capsules. Said dietary supplement may thus
contain from 10 to 100% by weight of the polyphenol-rich avocado
extract.
[0100] The composition of the present invention may be incorporated
directly and with no other modification in nutraceuticals, diet
products, notably hyper-protein products, or beverages by
techniques such as mixing, infusion, injection, blending,
absorption, kneading and spraying.
[0101] The modes of administration, posologies and optimal galenic
forms of the compounds and compositions of the invention may be
determined according to the criteria generally taken into account
in the establishment of a pharmaceutical treatment, in particular a
dermatological, cosmetic or veterinary treatment suitable for a
patient or an animal, such as for example the age or the body
weight of the patient or animal, the severity of the general
condition of the patient or animal, tolerance to the treatment,
noted side effects and skin type. Depending on the type of
administration desired, the active composition and/or compounds of
the invention may further include at least one pharmaceutically
acceptable carrier, in particular a dermatologically acceptable
carrier, or a cosmetically or nutraceutically acceptable carrier.
According to the first variant, a carrier suitable for
administration by external topical route is used. The composition
of the present invention may further include at least one
pharmaceutical or cosmetic adjuvant known to persons skilled in the
art, selected from thickeners, preservatives, fragrances,
colorants, chemical or mineral filters, moisturizing agents,
thermal waters, etc.
[0102] The composition including a polyphenol-rich avocado extract
having the indicated specifications is particularly intended for
cosmetic, pharmaceutical or dermatological use. The composition
will be formulated advantageously in the form of a preparation
suitable for topical administration.
[0103] The invention also has as an object the use of a
polyphenol-rich avocado extract for the manufacture of a cosmetic,
pharmaceutical, dermatological or nutraceutical composition or a
functional food.
[0104] Advantageously, the composition or extract of the present
invention is used in the prevention and/or treatment of disorders
or pathologies of the skin and/or mucous membranes and/or
appendages.
[0105] In particular, the composition or extract of the invention
is intended for the prevention and/or treatment of allergic,
inflammatory or irritative reactions or pathologies, or disorders
of the barrier or homeostasis of the skin, appendages (hair and
nails) and/or mucous membranes (gums, periodontium, genital
mucosa), whether immature, normal or mature/aged.
[0106] The expression "disorders of the barrier of the skin,
appendages and/or mucous membranes" refers to disorders intervening
at the level of the external layer of the skin.
[0107] The expression "disorders of the homeostasis of the skin,
appendages and/or mucous membranes" refers to disorders resulting
from the processes of cell renewal and equilibrium such as
psoriasis, diaper rash, atopic dermatitis, dry skin (xerosis),
dehydrated skin and photosensitive skin.
[0108] Advantageously, the composition or extract according to the
invention may be used for the prevention and/or treatment of
reactions, disorders or pathologies of the: [0109] skin, such as
rosacea or erythrocouperosis, psoriasis, vascular disorders, diaper
rash, atopic dermatitis, eczema, contact dermatitis, irritative
dermatitis, allergic dermatitis, seborrheic dermatitis (cradle
cap), sensitive skin, reactive skin, pruritus, dry skin (xerosis),
dehydrated skin, skin with redness, cutaneous erythema, aged or
photoaged skin, photosensitive skin, pigmented skin (melasma,
post-inflammatory pigmentation), skin with cellulitis, loose skin,
skin with stretch marks, scurf, chapping, bites, cracks, in
particular of the breasts, sunburn, inflammations due to rays of
all types, irritations by chemical, physical (for example tension
stress for pregnant women), bacteriological, fungal or viral,
parasitic (lice, scabies, tinea, mites, dermatophytes) or
radiological agents, or by a deficit in innate immunity
(antimicrobial peptides) or acquired immunity (cellular, humoral,
cytokines), and/or [0110] mucous membranes such as the gums and
periodontium subject to gingivitis (sensitive gums of newborns,
problems of hygiene, due the use of tobacco or other products),
periodontopathies, or genital mucosa presenting irritations of the
external or internal male or female genitalia, and/or [0111]
epithelial appendages such as the nails (breakable, fragile nails,
etc.) and hair (alopecia, dandruff, hirsutism, seborrheic
dermatitis, folliculitis) whether immature, normal or mature,
presenting in particular disorders of the scalp such as
androgenetic, acute, localized, cicatricial and congenital,
occipital in infants and greata alopecia (or pelade), due to
chemotherapy/radiotherapy or telogen effluvium, anagen effluvium,
pilar dystrophy, trichotillomania, tinea or oily or dry
dandruff.
[0112] The composition or extract of the present invention is also
used advantageously in the prevention and/or treatment of vascular
disorders, and may be used advantageously in hypopigmentation by
decreasing melanin and/or inhibiting tyrosinase, or as a healing
agent.
[0113] The invention also relates to a cosmetic care method for the
skin and/or appendages and/or mucous membranes, with a view to
improving the condition and/or appearance thereof, comprising the
administration of or consisting of administering a cosmetic
composition or an extract of the present invention.
[0114] In one embodiment of the cosmetic method of the invention,
the intended skin and/or appendagesand/or mucous membranes are
advantageously those that are sensitive, irritated or damaged by
the environment (UV rays, pollution), in particular sensitive
skin.
[0115] The cosmetic method of the invention is also characterized
in that the composition or extract is used as a moisturizing
product, or as a chronological or photo-induced anti-aging product
in the prevention of aging, and of photo-induced aging, or as a
slimming and/or anti-cellulitis product, or as an anti-aging or
anti-age spot product.
[0116] The expression "moisturizing product" refers to a product
that in particular makes it possible to prevent and/or treat
disorders of the barrier or homeostasis of the skin, appendages
and/or mucous membranes.
[0117] The expression "anti-age spot product" refers to a product
that makes it possible to reduce spots of pigmentation of the skin
and/or appendages and/or mucous membranes.
[0118] The expression "slimming and/or anti-cellulitis product"
refers to a product that makes it possible to improve the firmness,
elasticity or tonicity of the skin, and/or to fight the
accumulation of adipose tissue and skin with cellulitis.
EXAMPLE 1
[0119] Dried and defatted avocado flesh (oil cake) is suspended
under stirring at a concentration of 10% in a 60/40 (w/w)
ethanol/water mixture for 1 hour at room temperature. The residual
dry matter is separated from the liquid phase by filtration,
settling or centrifugation and the liquid phase thus obtained may
be filtered using filters of suitable pore size in order to obtain
a clear solution. The extract obtained has the following
characteristics: [0120] Dry extract: 2.56% [0121] Total sugars
(glucose, fructose, mannoheptulose, perseitol; HPLC): 19.5%/dry
[0122] Total polyphenols (Folin-Ciocalteu; gallic acid
equivalents): 14%/dry [0123] Proteins (Bradford assay):
6.25%/dry.
[0124] This extract has anti-radical activity, "in tubo" anti-DPPH
activity, for which the half maximal inhibitory concentration
(IC.sub.50) could be determined and is 111 .mu.g of dry extract,
which represents 10.6 .mu.g of polyphenols in the reaction
medium.
EXAMPLE 2
[0125] Dried and defatted avocado flesh (oil cake) is suspended
under stirring at a concentration of 10% in a 60/40 (w/w)
glycerol/water mixture for 1 hour at room temperature. The residual
dry matter is separated from the liquid phase by filtration,
settling or centrifugation and the liquid phase thus obtained may
be filtered using filters of suitable pore size in order to obtain
a clear solution. The extract obtained has the following
characteristics: [0126] Dry extract: 3.45% [0127] Total sugars
(glucose, fructose, mannoheptulose, perseitol; HPLC): 19%/dry
[0128] Total polyphenols (Folin-Ciocalteu; gallic acid
equivalents): 12%/dry [0129] Proteins (Bradford assay): 9%/dry.
[0130] This extract has anti-radical activity, "in tubo" anti-DPPH
activity, for which the half maximal inhibitory concentration
(IC.sub.50) could be determined and is 84 .mu.g of dry extract,
which represents 11.85 .mu.g of polyphenols in the reaction
medium.
EXAMPLE 3
[0131] Dried and defatted avocado flesh (oil cake) is suspended
under stirring at a concentration of 10% in a 60/40 (w/w)
propanediol/water mixture for 1 hour at room temperature. The
residual dry matter is separated from the liquid phase by
filtration, settling or centrifugation and the liquid phase thus
obtained may be filtered using filters of suitable pore size in
order to obtain a clear solution. The extract obtained has the
following characteristics: [0132] Dry extract: 2.74% [0133] Total
sugars (glucose, fructose, mannoheptulose, perseitol; HPLC):
22%/dry [0134] Total polyphenols (Folin-Ciocalteu; gallic acid
equivalents): 16%/dry [0135] Proteins (Bradford assay): 7%/dry.
[0136] This extract has anti-radical activity, "in tubo" anti-DPPH
activity, for which the half maximal inhibitory concentration
(IC.sub.50) could be determined and is 119.5 .mu.g of dry extract,
which represents 18.5 .mu.g of polyphenols in the reaction
medium.
EXAMPLE 4
Compositions for Topical Application
[0137] The inventors present below several compositions for topical
application. The polyphenol-rich extracts of avocado (Persea
gratissima and americana) may be incorporated in various cosmetic
products, such as cleansing water, oil-in-water emulsions,
water-in-oil emulsions, oils, milks, lotions, shampoos, foaming
products and sprays, whose compositions are presented below.
TABLE-US-00002 Cleansing water for sensitive skin Brand or INCI
name % Capryloyl glycine 0-1% Soda lye 0-1% Sequestrant 0-1%
Butylene glycol 1-5% Beta-carotene 0-2% Polyphenol-rich avocado
extract 0.01-10% Preservatives 0-1% PEG-32 1-5% PEG-7 palmcocoate
1-5% Zinc gluconate 0-1% Citric acid 0-1% Purified water q.s. to
100% Fragrance 0-1% Poloxamer 184 1-5%
TABLE-US-00003 Anti-aging emulsion Brand or INCI name % Liquid
isoparaffin 5-20% Isocetyl stearate 5-20% Al--Mg hydroxystearate
5-20% Abil WE 09 1-5% Glycerol 1-5% Vaseline oil 1-5% Micronized
zinc oxide 1-5% Butylene glycol 1-5% Retinol 0-1% Vitamin C 0-5%
Polyphenol-rich avocado extract 0.01-10% Isononyl isononanoate 1-5%
Beeswax 1-5% Sodium tartrate 1-5% Sodium chloride 0-5% Glycine 1-5%
Preservatives 0-1% Cholesterol 0-1% Phytosphingosine 0-1% Tartaric
acid 0-1% Purified water q.s. to 100%
TABLE-US-00004 Milk for dry, atopic skin Raw material/Brand or INCI
name % Sweet almond oil 1-5% Corn oil 1-5% Stearic acid 1-5%
C.sub.16-C.sub.18 cetyl acid 0-1% Antifoam 70414 0-1% Lauric
alcohol 11OE 1-5% PEG 300 monolaurate 0-1% Glycerol monoleate 0-1%
Glycerol monostearate 1-5% Vitamin B12 0-5% Polyphenol-rich avocado
extract 0.1-10% Preservatives 0-1% Citric acid 0-1% Trisodium
citrate 0-1% Purified water q.s. to 100% Fragrance 0-1% Peanut oil
1-5% Hydrogenated palm oil 1-5%
TABLE-US-00005 Cleansing cream Raw material/Brand or INCI name %
Purified water q.s. to 100% Arlatone 10-30% Cocoglucoside 5-20%
Hydroxypropyl guar 1-5% Capryloyl glycine 0-2% Preservatives 0-2%
Fragrance 0-1% Citric acid 0-1% Zinc PCA 0-1% Polyphenol-rich
avocado extract 0.01-10%
TABLE-US-00006 Antidandruff shampoo Raw material/Brand or INCI name
% Purified water q.s.to 100% Lauroamphoacetate 5-20% Cocoglucoside
5-20% PEG 6000 distearate 1-5% Preservatives 0-2% Vitamin F 0-5%
Piroctone olamine 0-2% Polyphenol-rich avocado extract 0.01-10%
Zinc pyrithione 0-1% pH adjuster 0-1% Sequestrant 0-1% Fragrance
0-1%
EXAMPLE 5
Compositions for Oral Administration
[0138] The avocado extracts are integrated into oral compositions,
in compositions enabling the administration of 50 to 200 mg of
polyphenol-rich avocado extract per day.
1/Anti-Stretch Marks Composition in the Form of Soft Capsules
TABLE-US-00007 [0139] Polyphenol-rich avocado extract 30 mg Awara
oil 60 mg Unsaponifiable-rich rapeseed oil 300 mg Vitamin of group
B (B1, B2, B3, B5, B6, B9, B12) q.s. to 100% RDA Tocotrienols q.s.
to 50% RDA Vitamin E Beeswax Soy lecithin Alimentary gelatin
Glycerin q.s. to 1 soft capsule
This composition is administered as four to six 500 mg capsules per
day.
2/Anti-Hair Loss Tablets
TABLE-US-00008 [0140] Polyphenol-rich avocado extract 25 mg Cereal
extracts (corn, buckwheat, millet, spelt) rich 200 mg in sulfur
amino acids Vitamin C q.s. to 50% RDA Fish cartilage
glycosaminoglycans 200 mg Glucidex IT 19 (compression agent) q.s.
to one 800 mg tablet
This composition is administered as five to eight tablets per
day.
3/Example of Slimming Powder Sticks
TABLE-US-00009 [0141] Polyphenol-rich avocado extract 100 mg
Polyphenol-rich tea extract 100 mg OPC-rich grape extract 50 mg
Plant beta-glucans 100 mg Xanthan gum 1 mg Sodium ascorbate 0.3 mg
Maltodextrin q.s. to 5 g
This composition is administered twice per day.
EXAMPLE 6
Biological Activities
[0142] 1. Anti-Inflammatory Effect
[0143] a. Anti-Inflammatory Activity on Keratinocytes
[0144] In the skin, the keratinocyte is one of the first cells
taking part in the initiation of the inflammatory reaction in
response to attack by the environment.
[0145] The "attacked" keratinocyte then releases cytokines which
induce a cascade of reactions involving the immune system.
[0146] Materials and Methods:
[0147] Human keratinocytes (NCTC-2544 cell line) were preincubated
or not (control) with the avocado extract concentrated in
polyphenols (AV) at concentrations of 0.005% and 0.01% (w/v of
active material) or the anti-inflammatory reference molecules
(10.sup.-7 M dexamethasone, 10.sup.-6 M indomethacin) for 24 hours.
The cells were then treated with 0.1 .mu.g/ml phorbol myristate
acetate (PMA) for 24 hours, always in the presence of AV or the
references.
[0148] At the end of the treatment, the quantities of interleukin 8
(IL8) and prostaglandin E2 (PGE2) secreted were measured by ELISA
in the culture supernatants.
[0149] The results were analyzed statistically by a Student's
t-test.
[0150] Results and Conclusion:
[0151] AV strongly and significantly inhibited the production of
the inflammatory mediators IL8 and PGE2 stimulated by PMA in
keratinocytes (Table 1).
[0152] The AV extract thus has an anti-inflammatory activity.
TABLE-US-00010 TABLE 1 Production of IL8 and PGE2 by keratinocytes
Inhibition IL8 (ng/ml) Control cells 0.1 .+-. 0.0 0.1 .mu.g/ml PMA
50.1 .+-. 1.8 10.sup.-7M Dexamethasone 7.4 .+-. 0.8 85% p <
0.001 0.005% AV 24.2 .+-. 1.4 52% p < 0.001 0.01% AV 13.7 .+-.
0.6 73% p < 0.001 PGE2 (ng/ml) Control cells 0.039 .+-. 0.0 0.1
.mu.g/ml PMA 138.4 .+-. 10.6 10.sup.-6M Indomethacin 0.039 .+-. 0.0
100% p < 0.001 0.005% AV 32.3 .+-. 3.1 77% p < 0.001 0.01% AV
10.1 .+-. 0.3 93% p < 0.001
[0153] b. Inhibition of the Production of Leukotriene B4 by
Neutrophils
[0154] Leukotriene B4 (LTB4) is a lipid inflammatory mediator
arising from the arachidonic acid pathway.
[0155] LTB4 is produced and released in large quantities by human
neutrophil granulocytes via the activation of the enzyme
5-lipoxygenase; it plays an essential role in the development of
cutaneous inflammatory reactions.
[0156] Materials and Methods:
[0157] Human neutrophils were preincubated for 15 minutes in the
presence of the avocado extract concentrated in polyphenols (AV) at
concentrations of 0.005%, 0.01% and 0.1% (w/v of active
material).
[0158] The cells were then stimulated by adding 1 mg/ml opsonized
zymosan.
[0159] After 10 minutes of incubation, the leukotriene B4 (LTB4)
released by the cells was assayed in the cell supernatants by an
ELISA technique.
[0160] The results were analyzed statistically by a Student's
t-test.
[0161] Results and Conclusion:
[0162] AV significantly inhibited the production of LTB4 induced by
opsonized zymosan on neutrophils (Table 2).
[0163] Thus, the AV extract modulates the inflammation induced by
the neutrophil and LTB4.
TABLE-US-00011 TABLE 2 Production of leukotriene B4 by neutrophils
LTB4 (pg/ml) Inhibition Control cells 31 .+-. 2.8 Stimulated cells
2900 .+-. 496.7 0.005% AV 2210 .+-. 779.3 -24% ns 0.01% AV 1737.5
.+-. 197.4 -40% p < 0.01 0.1% AV 1925 .+-. 434.9 -34% p <
0.05
[0164] C. Inhibition of Histamine Release
[0165] Mastocytes play an important role in allergic and
inflammatory reactions. In human, they are widely distributed in
connective tissue, such as the skin, serous mastocytes are the
sentinel cells of the local inflammatory response. Substance P, a
neuropeptide, acts on cutaneous mastocytes by inducing a rapid
release of histamine preformed in storage granules; this mechanism
of neurogenic inflammation is involved in various cutaneous
pathologies.
[0166] Materials and Methods:
[0167] Mastocytes were preincubated for 30 minutes in the presence
of the avocado extract concentrated in polyphenols (AV) at a
concentration of 0.01% (w/v of active material) or 10 mM calcium
(reference inhibitor of histamine release).
[0168] The mastocytes were then stimulated with 10 .mu.M substance
P for 15 minutes. At the end of incubation, the release of
histamine was quantified by ELISA.
[0169] The results were analyzed statistically by a Student's
t-test.
[0170] Results and Conclusion:
[0171] AV significantly inhibited the release of histamine by
mastocytes stimulated with substance P (Table 3).
[0172] The AV extract modulates neurogenic inflammation notably
related to histamine.
TABLE-US-00012 TABLE 3 Release of histamine by mastocytes Histamine
(ng/ml) Inhibition Control 20.1 .+-. 1.9 Substance P 142.5 .+-. 9.6
10 nM Calcium 21.8 .+-. 1.0 -85% p < 0.01 0.01% AV 112.5 .+-.
9.6 -21% p < 0.01
[0173] 2. Depigmenting Activity
[0174] The depigmenting effect of the AV extract was studied in two
distinct models: by evaluation of the production of melanin in
melanocytes; and by evaluation of the enzymatic activity of
tyrosinase, a key enzyme involved in the synthesis of melanin.
[0175] a. Inhibition of Melanogenesis
[0176] Materials and Methods:
[0177] Normal human epidermal melanocytes were cultured in the
presence of 10.sup.-7 M NDP-MSH (.alpha.-MSH analog; induction of
melanogenesis) and the avocado extract concentrated in polyphenols
(AV) at concentrations of 0.001% and 0.005% (w/v of active
material) or 0.25 mM kojic acid (reference).
[0178] After 240 hours of incubation, melanin was extracted from
the cells and quantified by spectrophotometry.
[0179] The results were analyzed statistically by a Student's
t-test.
[0180] Results and Conclusion:
[0181] AV significantly inhibited the production of melanin by
melanocytes stimulated by NDP-MSH (Table 4).
[0182] AV thus has a depigmenting effect.
TABLE-US-00013 TABLE 4 Melanin production by melanocytes Melanin
(.mu.l/ml) Control cells 24.7 .+-. 0.0 Stimulated control 30.4 .+-.
0.9 +19% p < 0.01 (NDP-MSH) Reference 9.6 .+-. 0.6 -32% p <
0.001 (kojic acid) 0.001% AV 26.2 .+-. 0.6 -14% p < 0.05 0.005%
AV 25.7 .+-. 0.5 -15% p < 0.05
[0183] b. Inhibition of Tyrosinase Activity
[0184] Materials and Methods:
[0185] The avocado extract concentrated in polyphenols (AV) and
various concentrations of kojic acid (reference) were preincubated
for 10 minutes at low temperature in the presence of tyrosinase
extracted from human melanocytes.
[0186] The substrate of the enzyme, 2 mM L-DOPA, was then
added.
[0187] After 1 hour of incubation at 37.degree. C., enzymatic
activity was evaluated by spectrophotometric measurement.
[0188] The results were analyzed statistically by a Student's
t-test.
[0189] Results and Conclusion:
[0190] AV significantly inhibited the enzymatic activity of
tyrosinase (Table 5). This result confirms the depigmenting effect
of this extract.
TABLE-US-00014 TABLE 5 Human tyrosinase activity Tyrosinase (U/ml)
Inhibition Control 182.7 0.0625 mM Kojic acid 166.8 -9% p < 0.05
0.25 mM Kojic acid 152.4 -17% p < 0.001 0.001% AV 165.5 -9% p
< 0.05 0.005% AV 164.7 -10% p < 0.05 0.01% AV 162.5 -11% p
< 0.01 0.05% AV 148.2 -19% p < 0.001 0.1% AV 142.4 -22% p
< 0.001
[0191] 3. Healing Activity: Stimulation of Markers of
Reepithelialization
[0192] The cicatrization mechanism in response to a wound involves
for epidermal repair a process of reepithelialization.
[0193] Cutaneous reepithelialization consists of the regeneration
by keratinocytes of an organized, pavimentous, stratified and
keratinized epithelium that covers the wound and that reconstitutes
a protective barrier.
[0194] The reepithelialization mechanism proceeds in three steps:
migration of keratinocytes, proliferation and maturation of the
epidermis.
[0195] The effect of the AV extract was studied on the gene
expression of markers involved in the keratinocyte migration step
during this reepithelialization process, laminin-5 and matrix
metalloproteinase-9 (MMP9).
[0196] Materials and Methods:
[0197] Normal human keratinocytes were incubated for 48 hours in
the presence of the avocado extract concentrated in polyphenols
(AV) at concentrations of 0.001% and 0.005% (w/v of active
material) or 5 ng/ml TGF.beta.1 (reference).
[0198] The gene expression of MMP9 and laminin-5 (subunit .gamma.2)
was studied by real-time RT-PCR.
[0199] The results were analyzed statistically by a one-factor
ANOVA followed by a Dunnett's test: ns (not significant) p>0.05;
*p<0.05; ***p<0.001.
[0200] Results and Conclusion:
[0201] AV significantly increased the expression of markers of
keratinocyte migration (Table 6).
[0202] This extract thus has a reepithelialization-activating
effect, in favor of a repairing, pro-healing effect.
TABLE-US-00015 TABLE 6 Gene expression of markers for keratinocyte
migration (relative quantity) Laminin-5 MMP9 Control cells 1.00
1.00 Reference 11.33 +1033% *** 19.84 +1884% *** (TGF.beta.1)
0.001% AV 1.62 +62% * 3.81 +281% *** 0.005% AV 1.51 +51% * 9.17
+817% ***
* * * * *