U.S. patent application number 12/058776 was filed with the patent office on 2008-10-09 for process for the preparation of ferutinine from ferula genus plants.
This patent application is currently assigned to INDENA S.P.A.. Invention is credited to Ezio Bombardelli, Aldo Cristoni, Gabriele Fontana, Enrico Mercalli.
Application Number | 20080248139 12/058776 |
Document ID | / |
Family ID | 33105046 |
Filed Date | 2008-10-09 |
United States Patent
Application |
20080248139 |
Kind Code |
A1 |
Bombardelli; Ezio ; et
al. |
October 9, 2008 |
PROCESS FOR THE PREPARATION OF FERUTININE FROM FERULA GENUS
PLANTS
Abstract
The invention relates to a process for the preparation of
ferutinine (Ia) from Ferula spp extracts comprising basic
hydrolysis of the extracts and treatment with p-pivaloyloxybenzoic
acid. The invention relates also to the use of the extracts and
ferutinine in the cosmetic and dermatological field.
##STR00001##
Inventors: |
Bombardelli; Ezio;
(Groppello Cairoli, IT) ; Fontana; Gabriele;
(Milano, IT) ; Cristoni; Aldo; (Milano, IT)
; Mercalli; Enrico; (Milano, IT) |
Correspondence
Address: |
YOUNG & THOMPSON
209 Madison Street, Suite 500
ALEXANDRIA
VA
22314
US
|
Assignee: |
INDENA S.P.A.
MILANO
IT
|
Family ID: |
33105046 |
Appl. No.: |
12/058776 |
Filed: |
March 31, 2008 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10551772 |
Oct 3, 2005 |
7371886 |
|
|
PCT/EP2004/003055 |
Mar 23, 2004 |
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12058776 |
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Current U.S.
Class: |
424/725 ;
514/510 |
Current CPC
Class: |
C07C 67/08 20130101;
A61Q 19/08 20130101; A61P 35/00 20180101; A61P 17/08 20180101; A61P
17/10 20180101; C07C 67/31 20130101; A61P 17/00 20180101; A61K
8/375 20130101; C07C 67/30 20130101; A61K 8/9789 20170801; C07C
2602/30 20170501; A61P 17/16 20180101; C07C 67/08 20130101; C07C
69/86 20130101; C07C 67/31 20130101; C07C 69/84 20130101 |
Class at
Publication: |
424/725 ;
514/510 |
International
Class: |
A61K 31/235 20060101
A61K031/235; A61K 36/23 20060101 A61K036/23; A61P 17/00 20060101
A61P017/00; A61Q 19/00 20060101 A61Q019/00 |
Foreign Application Data
Date |
Code |
Application Number |
Apr 4, 2003 |
IT |
MI2003A000661 |
Claims
1. A method of preparing a cosmetic or dermatological composition,
comprising: adding an effective amount of Ferula spp extract to an
acceptable excipient for a cosmetic or dermatological
composition.
2. The method according to claim 1, wherein the acceptable
excipient is selected from the group consisting of soy lecithin,
phospholipids, lauroylphosphatidylcoline and
myristoylphosphatidilcoline.
3. The method according to claim 1, further comprising:
incorporating the Ferula spp extract and excipient into one of an
emulsion and a transdermal plaster.
4. The method according to claim 1, wherein the cosmetic or
dermatological composition is in a form selected from the group
consisting of creams, gels and lotions.
5. The method according to claim 1, wherein the Ferula spp extract
is a Ferula hermonis extract.
6. The method according to claim 1, wherein the Ferula spp extract
is a Ferula communis extract.
7. A method of preparing a cosmetic or dermatological composition,
comprising: adding an effective amount of p-pivaloyloxyferutinine
to an acceptable excipient for a cosmetic or dermatological
composition.
8. The method according to claim 7, wherein the acceptable
excipient is selected from the group consisting of soy lecithin,
phospholipids, lauroylphosphatidylcoline and
myristoylphosphatidilcoline.
9. The method according to claim 7, further comprising:
incorporating the p-pivaloyloxyferutinine and excipient into one of
an emulsion and a transdermal plaster.
10. The method according to claim 7, wherein the cosmetic or
dermatological composition is in a form selected from the group
consisting of creams, gels and lotions.
Description
[0001] This application is divisional of U.S. application Ser. No.
10/551,772, filed Oct. 3, 2005, which is a 371 application of PCT
EP04/03005, filed Mar. 23, 2005.
FIELD OF THE INVENTION
[0002] The present invention relates to vegetable extracts from
Ferula spp and to a process for isolating ferutinine from said
extracts.
BACKGROUND OF THE INVENTION
[0003] Numerous Ferula genus plants contain terpenes with
estrogenic activity, known also as phytoestrogens, i.e. substances
which regulate hormonal functions and are apparently a valid
alternative to the use of synthetic hormones in the treatment of
pre-menstrual syndrome and disorders related to menopause and
aging. Extracts of some types of Ferula were used in ancient times
as contraceptives and in the treatment of impotence and menopausal
disorders. Recently, alcoholic extracts from Ferula asafoetida L.
have been disclosed (WO 0230438) as anticancer drugs.
[0004] The most abundant compounds in Ferula genus plants are
derivatives of jaeschkenadiol (II):
##STR00002##
in particular daucane esters having the general formula (I)
##STR00003##
[0005] Daucane esters are known compounds and are for example
disclosed in Phytochemistry, vol. 37, n.degree.3, pages 597-623,
1994. In the formula (I) R is a straight or branched, saturated or
unsaturated aliphatic acyl residue, or an optionally substituted
aromatic acyl residue. Examples of R groups are iso-valeroyl,
angeloyl, benzoyl, p-hydroxybenzyl, veratroyl or cinnammoyl.
[0006] Daucane esters from Ferula spp are estrogen modulators
similar to SERMs (selective estrogen receptor modulators); among
them, ferutinine (Ia) shows marked estrogenic activity, whereas the
others have a rather mild activity.
##STR00004##
[0007] In particular, ferutinine is an estrogen receptor alpha
agonist (.alpha.ER.alpha.) and an estrogen receptor beta
agonist/antagonist (ER.beta.). It has also been shown that
ferutinine has higher binding to estrogen receptors than
tamoxifen.
[0008] There is therefore the need to obtain ferutinine-enriched
extracts or optimise the extraction of ferutinine in the pure form
from plant materials containing its precursors.
[0009] A process comprising the hydrolysis of a daucane esters
whole extract to give crude jaeschkenadiol and the subsequent
re-esterification of jaeschkenadiol with suitably protected
p-hydroxybenzoic acid, for example with p-acetoxybenzoic acid, is
known from the literature (J. Org. Chem. USSR (Engl. Transl.); EN;
28; 10; 1992; 1666-1673). Nevertheless, this process gives rather
poor yields (about 45%), mainly due to competitive
transesterification reactions.
DETAILED DISCLOSURE OF THE INVENTION
[0010] It has now been found that the use of p-pivaloyloxybenzoic
acid as esterifying agent allows to avoid competitive reactions
responsible for low conversion yields.
[0011] Object of the present invention is therefore a process for
the preparation of ferutinine (Ia)
##STR00005##
[0012] which comprises the following steps:
[0013] a) extraction of daucane esters from Ferula spp;
[0014] b) basic hydrolysis of daucane esters to give jaeschkenadiol
(II)
##STR00006##
[0015] c) esterification of jaeschkenadiol (II) with
p-pivaloyloxybenzoic acid (III)
##STR00007##
[0016] to give p-pivaloyloxyferutinine (IV)
##STR00008##
[0017] d) hydrolysis of p-pivaloyloxyferutinine (IV) to
ferutinine.
[0018] "Daucane esters" means compounds of the general formula (I)
as defined above; said esters can be obtained by extraction of the
rhizomes or aerial parts of Ferula spp, preferably Ferula communis
and Ferula hermonis, with conventional methods, for example by
extraction with lower alcohols. Starting from Ferula hermonis
rhizomes containing ferutinine and jaeschkenadiol benzoic ester
(not easily separable by chromatography) in 1:1 ratio, pure
ferutinine can be isolated by extracting the roots with methanol,
treating the extract with 5% KOH and back-extracting the saponified
extract with aliphatic hydrocarbons, ethers, esters or chlorinated
solvents.
[0019] Alternatively, daucane esters can be obtained by extraction
with supercritical CO.sub.2 at temperatures ranging from 35 to
65.degree. C., preferably at 45.degree. C., and pressures ranging
from 200 to 260 bar, preferably at 245 bar. In the separator (or in
the separators) the temperature ranges from 25 to 45.degree. C. and
the pressure is of about 50 bar. In these experimental conditions
gummy materials, which make the recovery of the desired compounds
difficult, are not extracted. The residue can be directly submitted
to saponification according to what reported in the examples.
[0020] Jaeschkenadiol is esterified with p-pivaloylossibenzoic acid
and treated, in the same reaction solvent, with a base, preferably
a primary amine, more preferably ethylenediamine, to give pure
ferutinine. According to a preferred embodiment of the invention,
steps c) and d) are conveniently carried out in sequence without
recovery of intermediate p-pivaloyloxyferutinine.
[0021] A further object of the present invention is the cosmetic
and dermatological use of ferutinine, p-pivaloyloxyferutinine and
extracts, of Ferula spp, preferably Ferula communis and Ferula
hermonis extracts.
[0022] When applied on the skin, ferutinine and Ferula spp extracts
surprisingly proved able to increase collagen biosynthesis and to
exert a tonic, trophic and moisturizing action, thus giving
firmness and elasticity. Moreover, they reduce sebum secretion and
play a remarkable role in the control of hirsutism and face
virilization. Therefore, compositions containing ferutinine or
Ferula spp extracts can be used in the cosmetic or dermatological
field for the treatment of superficial or deep wrinkles or other
iunaesthetisms, as well as for the treatment of various acne and
seborrhea forms.
[0023] Ferutinine and Ferula spp extracts can be formulated in the
form of creams, gels and lotions in admixture with conventional
excipients, for example those described in Remington's
Pharmaceutical Sciences Handbook, XVII ed., Mack Pub., N.Y.,
U.S.A., preferably in the presence of soy lecithin or
phospholipids, such as lauroylphosphatidylcoline and
myristoylphosphatidilcoline, which can be incorporated in water/oil
and oil/water emulsions or in transdermal plasters.
[0024] The following examples illustrate the invention in greater
detail.
EXAMPLES
Example 1
Isolation of Jaeschkenadiol from Ferula hermonis Roots
[0025] 250 g of finely ground Ferula hermonis roots (particle size
distribution: 2 mm) are extracted by percolation with 1 l MeOH.
After maceration for two days and percolation of the solvent, the
operation is repeated (4.times.1 l), to obtain 112.2 g of methanol
extract (45%). Drug exhaustion is monitored by TLC (petroleum
ether/EtOAc 8/2, ferutinine Rf: 0.14).
[0026] The methanol extract is refluxed with 513 ml of a 10% KOH
methanol solution. After 1 h, TLC analysis (petroleum ether-EtOAc
8/2, ferutinine R.sub.f: 0.14; R.sub.f jaeschkenadiol: 0.31) shows
that the reaction is complete. After cooling, the reaction mixture
is diluted with water (500 ml) and extracted with petroleum ether
(4.times.500 ml). The combined petroleum ether phases are washed
with brine, dried and evaporated. The resulting semicrystalline
residue is washed with cold petroleum ether (refrigerator
temperature) to obtain 7.5 g of crystalline jaeschkenadiol. The
mother liquors are purified by chromatography (50 g silica gel,
petroleum-ether-EtOAc 95:5), to obtain .alpha.-bisabolol (870 mg),
a mixture of .alpha.-bisabolol and jaeschkenadiol, and pure
jaeschkenadiol (3.4 g after crystallization). The mixture of
jaeschkenadiol and .alpha.-bisabolol is pooled with the mother
liquor and chromatographed (50 g silica gel, petroleum ether-EtOAc
95:5), to obtain 1.95 g of crystalline jaeschkenadiol (overall
yield: 12.85 g, 5.1%).
[0027] The compound has the following chemical-physical and
spectroscopic properties:
[0028] IR spectrum (KBr, cm.sup.-1): 3339, 2965, 2905, 2875, 1470,
1375, 1047, 968, 858
[0029] Mass spectrum (C.I.)
[0030] M.sup.++1-H.sub.2O=221; M.sup.++1-2H.sub.2O=203
[0031] .sup.1H NMR spectrum (300 MHZ, CDCl.sub.3) .delta.: H9 5.43
m, H2 3.9 m, H14 1.78 s, H15 1.00 s, H12 0.95 d J 4.98, H13 0.91 d
J 5.13.
Example 2
Isolation of Jaeschkenadiol from Ferula communis Roots
[0032] 250 g of finely ground Ferula communis roots (particle size
distribution: 2 mm) are extracted by percolation using 1 l MeOH.
After maceration for two days and percolation of the solvent, the
operation is repeated (4.times.1 l); the methanol extracts are
concentrated to a volume equal to the weight of the ground roots
and the extract is added with 10% KOH (10 ml). The alkaline
solution is refluxed for 2 hours, then cooled and back-extracted
three times with 200 ml of n-hexane. Drug exhaustion is monitored
by TLC (petroleum ether/EtOAc 8/2).
[0033] The combined hexane phases are washed with brine, dried and
evaporated. The resulting semicrystalline residue is washed with
cold petroleum ether (refrigerator temperature), to obtain 3.5 g of
crystalline jaeschkenadiol. The mother liquor is purified according
to example 1. Yield: 0.8 g of jaeschkenadiol having the same
chemical-physical properties as that of example 1.
Example 3
Isolation of Jaeschkenadiol from Ferula communis Aerial Parts
[0034] 1 kg of finely ground Ferula communis aerial parts are
extracted with carbon dioxide at 45.degree. C. and 245 bar in an
apparatus for extraction with supercritical gases. In the separator
(or in the separators) temperature ranges from 25 to 45.degree. C.
and pressure is of about 50 bar. In these conditions gummy
materials that make it difficult to recover the desired compounds
are not extracted. The residue, which contains only lipophilic
compounds and water, is taken up with methanol and treated with
bases to hydrolyse jaeschkenadiol esters, as reported in examples 1
and 2. After purification, 5.1 g of pure compound having the same
characteristics of the product of example 1 is obtained.
Example 4
Synthesis of p-pivaloyloxybenzoic acid
[0035] 4-Hydroxybenzoic acid (114.5 g, 829 mmol) is dissolved under
stirring in pyridine (1.15 l), cooling to T<5.degree. C. on an
ice bath. The resulting solution is added with
4-dimethylaminopyridine (DMAP, 0.3 equivalents, 248.8 mmol, 30.4 g)
and pivaloyl chloride (3 equivalents, 2.487 mol, 300 g, 293.6 ml).
The solution is allowed to warm up to room temperature and left
under stirring for 2 h, then added with water (2.29 l) (exothermic
reaction: cool the solution in an ice bath) and left under stirring
for further 3 h.
[0036] The solution is poured into a separatory funnel and
extracted with CH.sub.2Cl.sub.2 (3.times.750 ml). The combined
methylene chloride phases are washed with 2 M H.sub.2SO.sub.4
(4.times.750 ml) and a saturated NaCl solution (1.times.1150 ml),
then dried over Na.sub.2SO.sub.4 (60 g).
[0037] The solution is filtered through paper filter and the
solvent is evaporated off under vacuum to obtain a residue which is
triturated with petroleum ether at 30.degree.-50.degree.
(3.times.400 ml), filtered by suction and dried under vacuum in a
static dryer at 45.degree. C. for 15 h. 130.5 g of product with the
following spectroscopic characteristics is obtained.
[0038] IR spectrum (KBr, cm.sup.-1): 3680, 2978, 2361, 1753, 1686,
1603, 1427, 1289, 1204, 1163, 1103.
[0039] Mass spectrum (C.I.): M.sup.++1=223
[0040] .sup.1H NMR spectrum (300 MHZ, D-DMSO) .delta. H3=7 8.00 d
J=8.48, H4=6 7.23 d J=8.55, CH3 1.34 s.
Example 5
Synthesis of Ferutinine from Jaeschkenadiol
[0041] Jaeschkenadiol (100 g, 419.5 mmol) is dissolved under
stirring at room temperature in CH.sub.2Cl.sub.2 (600 ml). The
resulting solution is added with p-pivaloyloxybenzoic acid (1.4
equivalents, 587.3 mmol, 130.5 g) and DMAP (0.3 equivalents, 125.9
mmol, 15.4 g). The solution is left under stirring for 10 min. to
complete reagents dissolution, then N,N'-dicyclohexylcarbodiimide
(DCC, 1.8 equivalents, 755.1 mmol, 155.8 g) is added. The reaction
is complete after 2 h.
[0042] The solution is concentrated to 2 volumes (200 ml) and
diluted with 5 volumes of CH.sub.3CN (500 ml), thereafter the
dicyclohexylurea precipitate is filtered off and washed with 5 more
volumes of CH.sub.3CN (2.times.250 ml). The combined organic phases
are poured into a separatory funnel, extracted with 10% w/v
Na.sub.2CO.sub.3 (2.times.250 ml) and with a NaCl saturated
solution (1.times.250 ml), then dried over Na.sub.2SO.sub.4 (100
g). Na.sub.2SO.sub.4 is filtered off and the solvent is evaporated
under vacuum to give 360 g of compound (IV), having the following
spectroscopic characteristics:
[0043] .sup.1H NMR (300 MHZ, CDCl.sub.3): .delta. 8.09 (d, J=9.0
Hz, H3'-H5'), 7.20 (d, J=8.7 Hz, H2'-H6'), 5.60 (brt, J=4.7 Hz,
H9), 5.35 (td, J=10.4-2.9 Hz, H6), 2.58 (dd, J=13.1-10.9 Hz, H7b),
2.34 (dd, J=14.1-2.3 Hz, H7a), 2.07 (m, H10), 2.04 (d, J=2.7 Hz,
H5), 1.97 (d, J=9.7 Hz, H2a), 1.89 (m, H11), 1.86 (s, H14), 1.63
(m, H2b), 1.57 (m, H3a), 1.41 (s, C(CH3)3), 1.30 (m, H3b), 1.14 (s,
H15), 0.99 (d, J=6.9 Hz, H12), 0.89 (d, J=6.7 Hz, H13).
[0044] Compound (IV) is dissolved under stirring at room
temperature in 2 l of CH.sub.2Cl.sub.2. The resulting solution is
added with ethylenediamine (10 equivalents, 280 ml). After 3 h the
reaction is complete. The solution is cooled to 0.degree. C.,
poured into a separatory funnel, then washed with 3 M
H.sub.2SO.sub.4 at 0.degree. C. (2.times.750 ml, exothermic
reaction) and a saturated NaCl solution (1.times.500 ml). The
organic phase is dried over Na.sub.2SO.sub.4 (100 g), filtered and
evaporated to dryness. The residue (230 g) is loaded onto a silica
gel column (2.5 kg) equilibrated with 5.8 l of a hexane:AcOEt=9:1
mixture and eluted with 70 l of the same mixture. The
product-containing fractions are pooled, the solvent is evaporated
off under vacuum and the product is dried in a static dryer at
45.degree. C. for 24 h.
[0045] 139 g (92.4%) of product having the following spectroscopic
characteristics is obtained:
[0046] IR spectrum (KBr, cm.sup.-1): 3410, 1686, 1655, 1608, 1593,
1560, 1279, 1165, 1099, 771.
[0047] Mass spectrum (C.I.): M.sup.++1-H.sub.2O=341
[0048] .sup.1H NMR spectrum (200 MHZ, CDCl.sub.3): .delta. H3'=7'
7.94 d J=8, 4'=6' 6.88 d J=8, H9 5.56 m, H2 5.23 dt J=11, H14 1.80
brs, H15 1.10 s, H13 0.94 d J=6.5, H12 0.82 d J=6.5.
Example 6
Preparation of a Ferula hermonis Extract
[0049] 1 kg of Ferula hermonis whole plant is extracted three times
with 5 volumes of acetone. The combined acetone extracts are
concentrated to 0.5 parts compared with the weight of the starting
biomass and diluted with 2 parts of water. The aqueous solution is
adjusted to pH 7.8 with diluted KOH, in the presence of hexane,
under strong stirring. The hexane phase is discarded, the aqueous
one is acidified to pH 5 and back-extracted with n-hexane. The
hexane phase that contains ferutinine is concentrated to dryness to
give 52 g of extract containing about 35% of ferutinine.
Example 7
Formulation Containing Ferutinine for the Treatment of Superficial
Wrinkles
[0050] Ferutinine is incorporated into a cream having the following
composition:
TABLE-US-00001 Ferutinine 0.20 g Carbomer 934 (Carbopol 934 P -
Goodrich) 0.60 g Propylene glycol 3.00 g Imidazolinylurea 0.30 g
Kathon CG 0.05 g Disodium EDTA 0.10 g PEG-5 soy sterols (Generol
122 E5 - Henkel) 2.00 g Octyldodecanol (Eutanol G - Henkel) 4.00 g
Wheat germ oil 4.00 g Silicone oil 350 cps 0.50 g Glycerylstearate
(Cutine GMS - Henkel) 7.00 g Polysorbate 60 (Tween 60 - ICI) 5.00 g
Tocopherol 0.20 g Ascorbyl palmitate 0.10 g 10% NaOH solution 2.00
g Perfume (186909 - Dragoco) 0.20 g Purified water up to 100.00
g
Example 8
Formulation Containing a Ferula hermonis Pure Extract with a
Ferutinine Content of 30% and a Jaeschkenadiol Benzoic Ester
Content of 20%
TABLE-US-00002 [0051] Ferula hermonis extract 0.5 g Carbomer 934
(Carbopol 934 P - Goodrich) 0.60 g Propylene glycol 3.00 g
Imidazolinylurea 0.30 g Kathon CG 0.05 g Disodium EDTA 0.10 g PEG-5
soy sterols (Generol 122 E5 - Henkel) 2.00 g Octyldodecanol
(Eutanol G - Henkel) 4.00 g Wheat germ oil 4.00 g Silicone oil 350
cps 0.50 g Glycerylstearate (Cutine GMS - Henkel) 7.00 g
Polysorbate 60 (Tween 60 - ICI) 5.00 g Tocopherol 0.20 g Ascorbyl
palmitate 0.10 g 10% NaOH solution 2.00 g Perfume (186909 -
Dragoco) 0.20 g Purified water up to 100.00 g
Example 9
Gel Containing Ferutinine
TABLE-US-00003 [0052] Ferutinine 0.30 g Imidazolinylurea 0.30 g
Methylparaben 0.20 g Hydroxyethylcellulose (Natrosol 250 HHX -
Aqualon) 2.00 g Purified water up to 100 ml
Example 10
Cosmetic Formulation containing Ferula spp Extract
TABLE-US-00004 [0053] Ferula hermonis extract 0.5 g
Imidazolinylurea 0.30 g Methylparaben 0.20 g Hydroxyethylcellulose
(Natrosol 250 HHX - Aqualon) 2.00 g Purified water up to 100 ml
Experimentation
Product Effectiveness
[0054] The effectiveness of the cream of example 7 was determined
in a double blind study with 40 female volunteers, of age ranging
from 39 to 56, evaluating the effects on skin elasticity and
firmness and on rugometry. The study was preceded by a seven days
conditioning period, wherein the subject had to refrain from the
use of moisturizing products, sun-creams and liquid make-ups and to
avoid tanning treatments and excessive exposition to UV rays.
[0055] The subjects were allowed to use conventional eye and lip
care products, face powders and non moisturizing soaps.
[0056] The subjects were randomly divided into two groups, one
treated with a placebo cream and one treated with the cream of
example 7. The creams were applied on the face in standardized
amount (0.5 g, i.e. 0.5 cm of cream coming out of the tube) twice a
day, morning and night. Before and after five weeks of treatment
the following measurements were carried out.
[0057] Before each measurement session all the subjects stayed for
thirty minutes in a climatic chamber at 23.degree. C. and 50% of
relative humidity. Each session comprised three measurements with
corneometer, three measurements with cutometer, and a silicon
impression of the periorbital area, on the skin areas indicated in
the following.
[0058] All the 40 subjects completed the study.
Cutometry
[0059] Cutometer is a commercially available device (Cutometer SEM
575, Courage & Khazaka, Germany) for measuring skin mechanical
properties in a non-invasive way. In more detail, it measures the
vertical deformation of the skin surface when subjected to a
negative pressure of 500 mm Hg through a 2 mm opening of a probe.
The length of skin penetration in the probe is optically measured
with 0.01 mm precision. The probe is connected with a computer that
registers skin deformation over time. From the resulting curve,
numerous variables can be extrapolated to evaluate skin elastic,
viscoelastic and viscous behaviour.
[0060] The following parametres were recorded:
[0061] immediate distension (Ue), measured at 0.1 seconds;
[0062] delayed distension (Uv);
[0063] final distension (Uf), measured at 10 seconds; and
[0064] immediate retraction (Ur).
[0065] The test was carried out using the cutometer on both
cheeks.
[0066] Significant variations were not observed in the placebo
group. Delayed distention (Uv) in the treated group significantly
decreased (16%, p<0.05) after 5 weeks treatment. This parameter
reflects skin viscoelastic properties and dermis behaviour. After 5
weeks, a significant change was also observed (-12%, p<0.05) in
Ue, which is mainly influenced by hydration and mechanic properties
of the corneum layer. The decrease in Uv and Ue, together with Ur
stability, shows increased skin firmness.
Corneometry
[0067] Soft and smooth skin appearance mostly depends on the
presence of an adequate amount of water in the corneous layer.
[0068] Corneometer is a commercially available device (Corneometer
CM 825 Combi 3, Courage & Khazaka, Germany) which measures
capacitance changes resulting from changes in skin hydration.
[0069] The test was carried out using the corneometer on both
cheeks.
[0070] After 5 weeks, a significant change in the treated group was
observed, in particular, skin hydration increased by 17.5%, while
in the placebo group it decreased by 3%.
Rugometry
[0071] Silicon impressions were carried out on subjects in the
seated position. The impressions (2.times.5 cm) were obtained at
the beginning and after 5 weeks, using "Silflo silicon impression
material" (available from Flexico, UK).
[0072] The impressions were then analysed with the Skin Image
Analyzer system using the Quantirides-Monaderm software, which
distinguishes cutaneous microrelief from median wrinkles and deep
wrinkles and calculate their number and depth; finally, the value
of total wrinkle area is obtained.
[0073] After 5 weeks, significant changes were observed in the
treated group. In particular, a 21.3% (p<0.05) decrease in the
wrinkle area was observed, whereas in the placebo group the
decrease was 0.4%.
[0074] Statistically significant decrease has been observed mainly
in the number and depth of median and deep wrinkles.
CONCLUSION
[0075] The study allowed to conclude that the cream of example 7
has good cosmetic activity in the treatment of skin with chrono-
and photoaging signs, as it increases skin firmness and hydration
and decreases mean wrinkled area, in particular deep micro- and
macrorugosities. Skin visibly appeared firmer and smoother.
* * * * *