Note: Descriptions are shown in the official language in which they were submitted.
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Title Q f the Invention
METHOD FOR THE TREATMENT OR PREVENTION
OF INTRINSICALLY AGED SKIN
S WITH RETINOIDS
Baekqrot-n~ of the I~ventio~
10Fiel~ of the ~n~ention
The present invention relates to a method for the treatment
or prevention of intrinsieally ~ehronolog~eally) aged ~kin by
the topical admini~tration of retinoids.
15De~er~ption of Prior Art
Cutaneous aqing results from two distinct and independent
processe~: intr~nsie aging an~ photoaging. Although tho ~kin
chanqes that oeeur in intrin~ie aging may invol~e qenetieally
programmed sene~eeneo mediated perhap~ by a~ yet unidentified
endogenous faetor~, an under~tanding of the elinieal
aiterations as well as ehanges in the estraeellular matri- and
moleeular biology is beginninq to emerge. Clinieally,
intrinsie aginq i~ eharaeterized by ha~ing a thin, atrophie
dermis, reduetion in subeutaneous adipose tissue and by the
presence of fine wrinkling. In addition, there appear~ an
unblemishe~ suraee with some ~eepening of s~in surfaee
markings an~ ~ome histopathologie and ultrastruetural
alterations a~ well. Compared to young-skin, intrinsieally
aged skin manifests it~ most pronouneed ehanges in the dermi~.
The ma~or ehanqe~ in the ela~tie tissue inelude an inerea~e in
the number as well a~ thieknesa of fibers, 108s of vertieal
fibers that insert into the basement membrane, an~ the presence
of large irregular fibroblast~ eontaining estensive rouqh
endoplasmie reticulum, eharaeteristie of elastogenesia.
'~
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. .
Conversly, photodamaged skin iJ characterized by
coarse wrinkl~nq and furrowing and the skin appears loose,
thickened and leathery. Photodamaged skin represents a
state of mild chronic inflammation. Histologically, in
photodamaged skin, there i~ e~cessive accumulation of
elastotic material in the upper dermis, basophiliC
degeneration and homogenization of the collagen fibers
accompanying the accumulation of
glycosaminoglycan-proteoglycan comple~es. Also, there
appears to be a loss of collasen bundles. Biochemically,
thera is a decrease in mature ~insoluble) collagen with a
concomitant increase in soluble collagen. Mature collagen
may be degraded by the chronic inflammatory infiltrato
produced by ultraviolet-B irradiation. However, in
lS intrinsically aged skin, mature collagen i8 moro stable
and resistant to enzymatic degradation, the bundles become
larger forming disoriented rope-like structure~. Also,
there is marked thickening or acanthosis in the epidermis
as well as the dermis and sebaceous glands become greatly
enlarged. Electronmicroscopic studies reveal that early
changes in photodamaged skin consist of an increase and
enlargement of the microfibrillar component of the elastic
fibers. In more advanced stages, the elastic fibers are
highly disorganized.
Concerning microcirculation, there is a loss of
vascular area in both intrinsic and photoaged skin, with
distinct differences between the two. Intrinsically-aged
skin e~periences a uniform reduction in the vasculaturs
with a progressive thinning of the vessel wall. The
vessels are not dilated or deranged and havo an
undisturbed superficial plesu~. In photoaged skin, the
vasculaturo i~ almost totally missing in some areas of the
dermis with the horizontal superficial ple~us almost
completely destroyed, ~hile in other areas the vesse1s
.
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become dilated and tortuous and are seen on the surface of
the skin as telangiectasias. In early photodamage, the
vessel wall actually thickens and then progressively thins.
Thus, there are distinct differences between
intrinsic aging and photoaging. Rligman, A. M. et al.,
"Cutaneous Aging: The Differences Between Intrinsic Aging and
Photoaging," Journal of Cutaneous aging and Cosmetic
Dermatology, Vol. 1, No. 1, pp. 5-12 (1988). These
differences are summarized in Table 1.
TABLE
Feature Intrinsic aging Photoaging
~1in; ~Al appearance SmDoth, unblemished ~ rrA~e ~llAr, le~thPry
Same ~PP~Pn;ng of skin ~lrrA~e with
rfA~e markings blotches, yellowing
Deep wrinkles
Some loss of elasticity Sign;f;~Ant loss of
elasticity
Epidermis fflin and viable Marked a~nthos;~
C~ 11lAr atypia
Elastic Tissue Increased, but almost Tremendbus increase,
normal f~ J~ ~r ~Les into
am~ hY.~ mass
~ollAgPn ~m~lf-s thick, ~i~nr;~nte~ f~arked decr~ase of
a and fibers
GAG~ PGs Slightly d~L~_sed Markedly increased
R~t;~llAr dermis Th;nn~r Th;~k~n~, elastosis
Fibrcblasts d~ ased, Fibroblæts increased,
inactive h~ f~Live
Mast cells de~L~ased, Mast cells mark~ly
no inflammation increased, mLxed
inflammatory infil-
- trate
P~r;llAry dermis No Grenz zone Solar elastosis with
Grenz zone
Micro~-~-lAtl-re Moderate loss Great loss, A~J~ r ~ 31
and t. ~1 An9i e~tatic
35 G~Gs glycosaninoglycans, PGs pL~Leoylycans
Tretinoin, all trans retinoic acid, has been described
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as being e~fect~e in the treatment of photoagea skin
(Weiss, J. J. et al., ~Topical Tretinoin Improves
Photoaged Skin,~ Journal of the American Medical
Association, Vol. 259, ~o. 4, pp. 527-532, (lg88). ~ittle
is known, however, of the effectiveness of retinoids in
the treatment or prQvention of intrinsically aged skin.
Vitamin-A acid has been suggested as being useful in
the treatment of wrinkles, sun damaged and sagging s~in.
Saline, C., ~Adventures in the Skin Trade.~ Philadelphia
Maga~ine, pp. 120-133 (1980). However, no method i~
described for the treatment of intrinsically aged skin by
the topical administration of retinoids.
Summary of the Invention
The present invention is directed to a method for the
treatment or prevention of intrinsically ~chronologically)
aged s~in by the topical administration of retinoids. The
retinoi~s employed may be any natural and/or synthetic
compound which po~sesses the biological activity of
vitamin A.
Detailed Description of the Invention
The invention relates to the use of retinoids for the
treatment or prevention of intrinsically ~chronologically)
aged skin.
Retinoids have been defined narrowly as comprisinq
simply ~itamin A (ret,inol) and its derivatives, such as
vitamin A aldehyde tretinal) and vitamin A acid (retinoic
acid), which comprise so-called natural retinoids.
However, subsequent research has resulted in a much larger
class of chemical compounds that sre deemed retinoids due
to their biological similarity to vitamin A and its
derivatives. Compounds useful in the ~res~nt invention
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include all natural and/or synthetic analog~ of v~tamin A
or retinol-lik~ compound~ wh~ch possess the biological
acti~ity of vitamin ~ in th~ skin. These include:
regulation of epithelial cell differentiation of
ketatinocyte~ ~n the epidermis; stimulation of new
collagen synthesis in the dermi~, and production of new
blood ~e~sels (angiogenesis). Accordingly, a~ used herein
for purpose~ of the present invention, the term ~retinoid~
will ~e understood to include any of the foregoing
compound~. Esample~ of suitable retinoid~ in the present
in~ention are set forth in Table 2, although it will bo
un~er~tood that the invention i~ not limited thereto.
Table 2
all-tran~-retinoic acid
13-ci~-retinoic acid
.
(all-E)-9-(4-methosy-2,3,6-
trimethylphenyl)-3,7-dimethyl-2,4,6,8-
nonatetraenoic acid ethyl ester
(all-E)-9-(4-methosy-2,3,6-
25trimethylphenyl-3,7-dimethyl-2,4,6,8-
nonatetraenoic acid
N-ethyl-9-(4-methosy-2,3,6-trimethyl-
phenyl)-3,7-dimethyl-2,4,6,8-
~onatetraenamide
(E,E)-9-(2,6-dichloro-4-methosy-3-
methylphenyl)-3,7-dimethyl-2,4,6,8-
nonatetraenoic acid ethyl ester
Table 2 (continued)
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7,8-didehydroretinoic acid
(E,E)-4-~2-methyl-4-(2,6,6-trimethyl-
1-cyclohesen-1-yl)-1,3-butadienyl]benzoie acid
(E)-4-t4-methyl-6-(2,6,6-trimethyl-
l-cyclohesen-l-yl)-1,3,5-hesa-
trienyl]benzoic acid
tall-E)-3~7-dimethyl-(3-thienyl)
2,4,6,8-nonatetraenoic acid
(E,E,E)-3-methyl-7-(5,6,7,8-tetrahydro-
lS 5,5,8,8-tetramethyl-2-naphthalenyl)-
2,4,6-octatrienoic acid
(E)-6-12-(2,6,6-trimethyl-1-cyclohesen-
l-yl)ethenyl~-2-naphthalenecarbosylic acid
(E,E,E)-7-(2,3-dihydro-1,1,3,3-tetra-
methyl-lH-inden-5-yl)-3-methyl-
2,4,6-octatrienoic acid
(E)-4-t2-(2,3-dihydro-},1,3,3-tetramethyl-
lH-inden-S-yl)-l-propenyl]benzoic acid
(E)-4-t2-(5,6,7,8-tetrahydro-5,5,8,8-
tetramethyl-2-naphthalenyl-1-
propenyl]benzoic acid
(E)-4-[2-(5,6,7,8-tetrahydro-3-methyl-
5,5,8,8-tetramethyl-2-naphthalenyl-1-
propenyl]benzoic acid
Table 2 (continued)
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(E)-1,2,3,4-tetrahydro-1,1,4,4-tetramethyl-
6-(1-methy}-2-phenylethenyl)naphthalene
56-(1,2,3,4-tetrahydro-1,1,4,4-tetramethyl-
6-naphthyl)-2-naphthalenecarbosylic acid
(E)-6-t2-(4-(ethylsulfonyl)phenyll-1-
methylethenyll-1,2,3,4-tetrahydro-
101,1,4,4-tetramethylnaphthalene
4-t(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-
2-naphthalenyl)ethynyl]benzoic acid
15(E) -2-(1,1,4,4-tetramethyl-1,2,3,4-tetra-
hydronaphth-7-yl)-1-t4-tetrazol-
5-yl)phenyll-1-propene
(E) -4- E 2-(S,6,7,8-tetrahydro-7-hydro~y-
205,5,8,8-tetramethyl-2-naphthalenyl)-
l-propenyllbenzyl alcohol
Included among the compounds which can be employed are
the pharmaceutically acceptable addition salts and esters
of the retinoids such as the palmitates etc. Also
encompassed within the term ~retinoid~ are geometric and
stereo~somers of the retinoids.
A pharmacological composition containing a retinoid a~
the active ingredient in intimate admisture with a
pharmaceutical carrier can be prepared according to
conventional pharmaceutical compounding techniques, such
as, for esample, those known for topical application of
all-trans-retinoid acid. The carrier may take a wide
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variety of physical forms such as, for e~ample, creams,
dressinqs, gels, lotion~, ointments or li~uids. The
retinoid will be present ~n the composition in an amount
from about 0.00001~ by weight to about 0.2% by weight,
depending on the potency of the retinoid. A suitable
topical retinoid preparation in a gel ~ehicle i~ Retin-A~,
which contain~ 0.01~ to 0.1~ by weight of the activ~
ingredient, ~produced by Ortho Pharmaceutical
Corporation).
The following esample describe~ the invention in
greater particularity, and is intended to be a way of
illustrating but not limiting the invention.
EXAMP~E 1
Lifetime Application of all-trans-Retinoic Acid to Albino
Hairless Mice.
Materials an~ Metho~s
Female Albino hairless mice (Skh-hairless-l), age 8-11
weeks, were treated topically, three times a week, for the
remainder of their life span as follows:
Group 1. 0.025% all-trans-retinoic acid
(100 ~1) 12 mice
Group 2. Creaml~ehicle (100 ~1) 12 mice
Group 3. Untreated 12 mice
The mice were esamined monthly for skin changes and
possible weight loss that might indicate a to~ic effect.
Toward the end of the study, mice were photographed to
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show the condition of the skin. A few mice were
sacrificed at about 70, 80 and 90 weeks, with dorsal skin
taken for histochemical evaluation. The remainder were
allowed to live out their life span which ranged from
91-106 weeks and were biopsied when moribound, if
possible. The histochemical stains were: ~&E for general
histology, Luna's for elastic fibers, Yan Gieson's for
collagen and Mowry' 8 for proteoglycans.
Res~llt~
A. To~icity
1. There was no difference in the weights of mice
with~n the three groups.
2. There was no difference in mortality within the
three groups.
3. At biopsy and necropsy there was no indication
that the mice suffered any adverse or unusual
effects.
Therefore: treatment with either all-trans-retinoic acid
or vehicle over a lifetime produced no detectable tosic
effects.
B. Clinical Observation~
Mice treated with all-trans-retinoic acid had thinner,
pinker, more uniform skin than the controls. They more
closely resembled far younger, untreated mice. Vehicle
controls had thicker, yellower skin. Untreated controls
had e~tremely thick, sagging, yellowed skin, typical of
such aged mice (Figure 1).
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Figure 1. is a photograph which illustrates the
results of lifetime application of vehicle or
all-trans-retinoic acid to female albino hairles (Skh-l)
mice. t Yehicle, tt - untreated, ttt -
all-trans-retinoic acid. Mice were 91-106 weeks of age at
photoqraph. All-trans-retinoic acid mice haa thinner,
pinker, more uniform skin than controls or untreated
mice. They resemble young mice. Vehicle control mice had
thicker, yellow skin. Untreated mice had very thick,
sagging, yellowed skin.
C. Histochemical Observations
~&E for General Histology (See Figures 2, 3 and 4)
Figure 2. is a picture of a normal old mouse, at 80
weeks of age. H & E stain.
Figure 3 is a picture which illustrates cream vehicle
mice at 85 weeks of age.
Figure 4 is a picture which illustrates mice treated
with all-trans-retinoic acid. (~ifetime of treatment~.
Hyperplastic epidermis and strongly granular layer of
appro~imately 4 cells. Cells are plump, cytologically
normal and have abundant cytoplasm and an abundance of new
blood vessel~. Sample wa~ taken from a mouse g5 weeks of
age.
In all-trans-retinoic acid-treated specimens, the
epidermis wa~ hyperplastic (about 8 cell layers) including
a strongly granular layer of about 4 cell layers. The
1 33565 1
cell~ were plump, cytologically normal and had an abundant
cytoplasm. Control specimens had the usual 2-4
compressed-looking cells. Blood ~e~sel~ were ~eadily seen
in the all-trans-ret~noic acid group, unl~ke in the
controls. Areas of new collagen were apparent by the
parallel array of the bundles and the presence of numerou~
large fibroblasts. These regions were free of
inflammation despite a mild inflammatory infiltrate in the
mid-dermis. No repair areas were seen in the control~ and
dermal inflammation was present but variable from specimen
to spec~men.
r~lna~ Stain for ~lastic F~ber~
Thi~ stain confirmed the presence of new collagen in
the sub-epi~ermal dermi~ of Retin-A~-treated mice. The~e
areas, termed repair zones, are identified by elastic
fiber~ at their lower border, having been displaced
downward by the deposition of new collagen. Repair zones
were intermittent acros~ the specimen, unlike in
photodamaged skin where they are continuou~. In control~,
no repair zones were found (Figure 2). In addition, mice
treated with all-trans-retinoic acid appeared to have an
2S increase~ amount of new elastic fibers (Figure 4).
Van Gies~^s Stain fo~ Colla~pn
Collagen in the repair zone~ stained mainly as normal,
mature collagen.
MQwrY's Stain for GlYcosamino- an~ proteoqlycan~
There was little or no increase in dermal
~5 glycosaminogly~ans ~r proteoglycan~ in any of the groups
1 335651
escept for intermittent depo~its at the dermal-epidermal
junction. These deposits are typically found in aging
mice. Fewer ~eposits were found in the retinoid treated
group.
EXAMP~E 2
Five, healthy white women, ages 7~ to 85 receive~
treatment or control~ on their upper lateral thigh once
daily. One s~d~ recei~ed Purpose Cream~ (control), the
other side ha~ Retin-A~ 0.05~ applied. The duration of
the study is for one year.
Clinical results eight months into the study are as
lS follows:
Some improvement on both sides was noted, but it was
e~eedl~gly better on the Retin-A si~e. At tb- start of
the esperlment the sk~n was wrinkled, rough, dry, loose,
etc. With ~etin-A after S months the s~in was smooth,
tighter, less wrinklea and firmer. Thero was som- initial
irritation with Retin-A but the skin accommodated.
Skin biopsies were taken from another elderly female 6
months into the study.
A histoloqical e~amination of the Retin-A cream treated
skin showed acanthosis, an increased granular layer, a
compact horny layer~ less epidermal atypia and dysplasis,
more dermal cells (fibroblasts and lymphocytes) with the
morpholoqy of metabolic hyperactivity. On the control
side, chanqes were slight.
Retin-A~ was clearly~shown to be efective against
i~trinslc~l~y agod ski~ esght month~ into the study.
