CA2035459A1

CA2035459A1 - Method for treating skin to repair the effects of photoaging

Info

Publication number: CA2035459A1
Application number: CA002035459A
Authority: CA
Inventors: Neil F. Haley; Xina Nair; Gerard J. Gendimenico
Original assignee: Individual
Current assignee: Eastman Kodak Co
Priority date: 1989-07-25
Filing date: 1990-07-19
Publication date: 1991-01-26
Also published as: JPH04500824A; EP0436010A1; WO1991001128A1; HUT62193A; HU906129D0; KR920700612A

Abstract

The effects of photoaging or sun damage of skin including loss of collagen fibers and deterioration of small blood vessels in the dermis of the skin are repaired by applying topically to the epidermis effective amounts of a compound having structure (I), wherein R1, R2, R3, R4 and R5 are independently selected from the group consisting of H, Cl, OR6, straight or branched alkyl of 1 to 10 carbon atoms, NO2, COOR6, CN, OR6, NR6R7, NR6C(=S)NR7R8, NR6COR7, SO2NR6R7, CH(CH3)COOH, CONR6R7, COR6, OCONR6R7, NR6COONR7, R9OR6, NR6SO2R7, Si(CH3)3, and NR6CONR7R8, R3 together with R4 forms a benzo ring or taken together with R2 forms a benzo or tetrahydrobenzo ring or together with R2 and R1 forms a (1) moiety or together with R2 forms a (2) moiety or R2 together with R1 forms a benzo ring or R2 together with R3 forms a (3) or (4) or (5) or (6) moiety, or R1 is independently selected from the group consisting of (7), (8) moiety, R6, R7 and R8 are independently selected from the group consisting of straight or branched alkyl containing from 1 to 10 carbon atoms, aryl containing from 6 to 10 carbon atoms and hydrogen, and R9 is alkylene of 1 to 6 carbon atoms, and iron carbonyl complexes thereof, to an area of the skin in an amount sufficient to repair the effects of elastosis in the skin. Lines and wrinkles due to aging are reduced and prevented.

Description

W o 91/01128 PCT/US90/0405 SKIN COMPOSITION TO REPAIR THE EFFECTS OF PHOTOAGING

Cross-Reference to Related Applications This application is a Continuation-In-Part of copending U.S. Application Serial No. 384,949 fiied on July ~5, 1989.
ield of the Inven~iQn This invention relates to a method of repairing the eîîects oî aging of the skin, particularly human facial skin, by topical application of specific polyene compositions~
R~?~C ~ 5~L5~ Inven~Q~
~ ;c~ssive exposure of hu~an skin to sunlight caus~3 da.~e, r~sultin~ i~ a nu~ber of gross cu~aneou~ change3. ~hese i~ciuàe wri~kling, lea.her-iness, yellowing, looseness, roughness, dryness, mottling (pigment spots~ and benign and malignant skin tumors. A person exhibiting these signs looks prematurely aged. Photoaging is most prominent in light skinned persons who brown easily and tan poorly. The effects of sunlight are cumulative. As a result, this sunlight-induced skin damage has been referred to as photoaging.
The majority of signs of photoaging can be prevented by judicious use of topically applied sunscreen agents. It is important to use sunscreens early in life, for example, as soon as a child begins to spend a significant amount of time outdoors.
Many persons will neglect to use sunscreens, and worse, some will intentionally overexpose them-selves to natural or artificial sunlight to benefit from cosmetic attributes of tanned skin. Later in life~ they will seek medical care in the hope of alleviating the skin damage, for instance, by under-going cosmetic surgery.

~ r. ?~ 2-A pharmaceutical composition that can promote the repair of photoaged skin is an alterna-tive treatment to patients who neglect to use sun-screens and do not pr~rer cos~e ic sur~o-~v. Tooi-cally applied all-trans .et-;noic ac-d is reported to impro'~'~? ~ ? ~??~`a~ à~ ? `~ ` 1`- ?~ '`?'~ IC
double-blind studies. ~he ~eile,iciai c:.~nges were clinically signi~ica~ o ~ inV~S.l.-~?._~rS and the patients. These included e~^^ac~ment o` `ine wrin~les, r~duc?d s.';..~. SOtl~``..'.''~S. irx~u ~ pin.~s-.ing of th~? s;;~.- a~d l~ ?.~ ?~ ?~ ?~ ns ~len.igines~ solar-;ndu,~d ~`.?~
In tha d~ 27 ~_h~ J~t~?'~
th2t ~
lS cha~ , v~ v~iiin~
xerosis, and miid scaling. _lovon c^ :~o patients needed potent topical ste-oi A3 to ~ te the dermatitis. Three patients withdrew from the study because of the severity of retinoid-induced dermati-tis.
It has been sought to provide a method for the treatment of photoaged skin, but without the adverse effects of dermatitis as noted with all-trans retinoic acid treatment.
The use of 13-cis- ~ ino c _cid as a treat-ment for the adverse effects of elastosis is des-cribed in Australian Patent AU-A-830271~7 by ~offman-La Roche and Furopean Patent Appiication Publication 025333.
In U.S. Patent 4~5~,696 certain polyenes are described as being usefui in treating inflamma-tory or allergic conditions~ These conditions, of course are far afield of pho.oaging and ~aterials useful for the treatment of inflammatory conditions are not expected to be usefui in the treatment of photoa~ing and vice versa.

WO 91/01128 PCr/US90/040~;2 --3~ r ~ ~;
In U.S. Patent 4,603,146 it is disclosed ~
that all-trans retinoic acid (retin A~ is useful in the treatment of photoaging. However, this treatment also results in great irri~tion to the skin, which severely limits its use.rul.nAess.
Summarv o~ h~
The present invC~ntion rela.es .o the use of specific ~lyenes in re~ai-i~ he ~gi..~ changes of exposed areas or th~o sXin, ~s?ociall~ he face.
These pol~-enes ...~-ar~ ~.nd -~ ~-;e `-~ 'ss~ of collagen ribers ~nd deterio-~ n e ~m~:l b1Ae~d VeSSe1 S ~ r~r ~ t~ ,r,/~ ,Q
patient~

repairing o-,- 3un-~dinaae~ eom~-~ S~.g topicaily administering a com?ound h2';i?.-^ the s zs ure:

H C C~ ClH3 CH3 5`I~ \ ~R3 I' `O' ~ -' `' ~T' \R~
\-~ \C~ R
wherein Rl, R2, R3~ R4 and R5 are inde-vendently selected from ~he grouv consisting of ~, Ci, OR6, straight or branched alkyl of l to lO
carbon atoms, N02, COOR6, CN~ OR~ NR~R,~
NR6c(=s)NR7Rg~ NR6CR7 S~2h~6 7' C~(C~3)COO~, CONR6R7, COR6, OCONR6R7, 6 7- RgR6- NR6SiR7.
Si(C~3)3, and NR6CONR7R8, R3 together with R~ forms a benzo ring or taken togethe with R~ rorms a ben~o or tetrahydrobenzo ring or together with R2 and R

WOgl/01128 PCT/US90/04052 ~,J~v`;~ ~ forms a:
I
1~ ~
moiety or together with R2 forms a lo mQie~ - '~2 ~ogether with Rl ~orms a ~e.._o ring or ~ ~ogether with R3 forms a lS I or \- or ~~- or -S~.
_o' ~ -C~2 -C~
moiety, or Rl is independently selected from the 20 group consisting of CoN~O~
N\ ~S

moiety, R6, R7 and R8 are independently selected from the group consisting of straight or 30 branched alkyl containing from 1 to 10 carbon atoms, aryl containing from 6 to 10 carbon atoms and hydrogen, and Rg is alkylene of 1 to 6 carbon atoms, and iron carbonyl comp exes thereof, to an area of the skin in an amount sufficient to repair the effects of elastosis in the WO91/01128 PCT/US90/~4,0,52~ ~
h., i~,_^.,, skin. Lines and wrinkles due to aging are reduced and prevented.
The polyenes may be applied to the skin in any non-toxic, dermatologically acceptable vehicle, preferably a non-volatile emollient or lubricating vehicle in varied concen.rations which i3 more fully described hereinbelow.
Detailed Description of the ~ree~lss~ m~n~
The treatment o skin with ~he polyenes of the present invention moderate and retar~ the aging changes in the skin to both the dermis and the epidermis. As a~e and exposure to sunlight increases, the skin's cells di~ide at a slower rate~
The ~ic';ness Oc .he epide~mis de~rease~ and the horny iayer wnich pro-ects against water loss sheds cells in large groups, resulting in rough, dry and scaling skin~ The cells whlch make up the fiber of the dermis become smaller with increasing age with a loss of collagen fibers. The degradation of these fibers contributes to wrinkling and`loss of elasti-city.
The polyene compounds useful in the present invention have the structure:
R
C~ C~ o \ ~ \ ~R3 ~3C\ ~CH3 l 3 . l 3 ~ I O
o~ ~t~ ~R2 \~ \C~ R
wherein Rl, R2~ R3~ R4 and R5 are independently selected from the group consisting of ~, Cl, OR6, straight or branched alkyl of l to 10 carbon atoms, for which examples are provided hereinbelow, N02, COOR6, CN, OR6, NR6R7, NR6C(=S)NR7R8 . ~'R6CR7 ~ S2NR6R7 ' r 9 --6--CH(CH3)C00~, CONR6R7, COR6, OCONR6R7, NR6COONR7, R90R6, NRSS02R7, Si(C~3)3, and NR6CONR~R8, R3 t~gether ;~i~h ?~4 ~-ms a b~r._o ring S or taken t~ h~ h ~2 _o_ms ~ ben~o or tetr~ - ' ' ?~ . ' forms a:

! ~T
.

moiety c~ o.-s~;ler ~l s ~ ms a ~;1-, R
,`~

\ "
moiety or R2 together with Rl forms a ben~o ring 2 or R2 together with R3 forms a `I -` -` -s o~ _o~ -c~ -c~
moiety, or 25Rl is independenti~ selected frQm the group consisting of d i -CON~
N~ ,S

moiety, R6, R7 and R~ are independently selected from the group consisting of straight or branched alkyl cont~ining f:^~ l to iO ca-bon atoms, WO91/01128 PCT/US90/040~2 7 ~;Jc) 5 ~r ~ 9 for which examples are provided hereinbelow, and aryl containing from 6 to lO carbon atoms and hydrogen, for which examples are ~ro~ided hereinbelow, and ?~9 is al~ylene o- l to 6 carbon atoms, such as ~e hjl~-.e, ?r~ lenq~ bu~ylenQ, ~rimethylene, etc.
and iron carbonyl comrlexes thereof such as CO
C~l,C0 l4 ;~ R3 ~ ~ 2 ~~3 ~ Rl 15 ~ ^ù~ ;a~p'~s of al~yl o l .o ~ oar~o~ oi~s .~re ~ hyl, butyl, pentyl, octyl, ethyl, terti_ry-butyl, ben2yl, isopropyl, chloroethyl, chloropropyl, hydroxypropyl, carboxyethyl, carboxymethyl, phenynyl, cyanoethyl, and 2-ethylhexyl and aryl of 6 to lO carbon atoms are exemplified by phenyl and napthyl.
The method of preparing these polyenes is well known and is generally described in U.S. Patent 4,595,696(incorporated herein by reference).
Generally, the co~pounds are ormed by leaction of polyene acids with acetic anhydride, boron trifluoride, oxalkylene chloride, phosphorous trichloride or thionyl chloride and then further treated with phenolic compounds~
Useful polyenes within the scope of the present invention include those with the following structures:
I.

C~ ~CH3. ~ ~ 3 C~3 ._.

PCr/US90/04052 J ~ '~ v ~ -8-II .

3 \.~ 3. 1 3 . 1 3 e ~ CH3 \ ~ \CH

III .

H3C~ ~CX3 ~CH3 CX3 0 ~ ~ ,coOC:~3 ~ `o' ' \ / \CH
I'\~ .

1~ \o~ ~ ~e~O~

V.
I~ \o~ f~-~

25 Vl.
~3l~ ~oc~3~ ~lc~3./.~.~f~3~e\ ~

VII .

H3 C, ~C~3 ClH3 ClH3 e O ' ;~ I

\-~ \C~3 Cl WO 91/01~28 _9_ VIII . ~ ;' `' " -H3C, /C~3 l 3 l 3 o ~I~ ,I

\./ \CH

IX .

~3C, C~ Cl 3 Cl 3 IJ ~ t~ ~I

X~ ~ C.~3 E13C,,C~3 1 3 1 3 ~ ~ t '~--\

XI .
~-\

XII .

I' `O' ~ ~ ~o/ ` ~I
\ / \CH

PCl /1 1S90/04052 ~, 5 ,/~ l0--XIII .

3~ /3,~ 3"~ o\ 5I

s ~, ~, XIV .
lc~3 ~ , Co2EI

X~ ~
~ r r~ CrI,~ cr o ~` ~
J\O~
`. `c~3 20 XVI.

1~ ,0, ~ ~o,O~ ~I

\-~ \C}I N~,OC}I3 XVI I .
,3~ .,f~ o,O~ ~I

\ ~ \CH

WO 91/01128 PCI`/US90/04052 XVIII . ~ ;.~ cj " ''-'`
CO O

3 \ ,~ ~3, 1 3 ~ I~ 3 ! ~--?
\ ~'\ c~3 XIX .

\ " ~ - C ~ ,~ 1l \ .. _ _ ~ 3 15 ;~X.
~3C, ,,CE3 f 3 f~3 1l I~ `0~ 3 2 O ~ o~
20~-~ \C~I
XXI .
~3C, ~CH3 l 3 l 3 ~ I~ ,O~
\." \C~I

XXII .

~3C C~ ~ ic~3 C~3 I~^~ ,C0CH3 \~ \CH

WO 91/01128 PCT'/US90/040;2 ,'. r~ 2 XXIII .
1~ H CH CH O ~: \
3C\.,C3 13 13~ I O
I' `O' ~ ' `' ~T' \-/ \CH CON~2 XXIV .
~3C, "C~3 fFI3 CX3 Q ~-, ,~(C~3)C~)C; 3 lo I O ~ `Q~
\ ,~ \

XXV

~C\ ~C~3 1C~3 C;i3 Q i~`
`' ~T' \-~ \CH N~2 XXVI .

3.~ 3.~ 1 . f~ I O
O ~ -' `o~ ~t~
\-~ \CH OCH3 25 XXVII.
H3C, ,CH3 f~3 f~3 ~ t~ `O
O ~ -' `o~ c~
CH

XXVIII .
~I C C~3 C
3 , ,CH3 1 . 1 3 1! I O
O ~ -' `o~ ~t~
\ ~ \CH C0N(CH3)2 W09ltOI~28 PCT/US90/04052 22I2, ;`~
H3C CH lCH3 CH3 0 \ ~ \CH

XX2.
~3C, ,C~ lC~3 1~3 o I~
lo I~ `9' ~ f ~1 \~/ \C~ N~ 3~,0C~3 22XI.
15 ~3~,",C~3 C~3!C~3 !I t ;1 I O ~ ' \i~ c~, \-~ \CH
XX2II.
3 \.~ ,~ f~ ~ I O

`.~ `c XXXIII.
CH 3 CH3 C~ 8 ~^`
I' `O' ~ ' `' ~ ' ~-COC~3 \.~ \CH

XXXIV.

~3C, ,C~ ClH3 lC 3 ~o I `O

WO 91/01128 PCI`/US90/04052 .
~ r q ~ 1 4 h ~J ~ XXXV.

~3C\ /C 3 1 3 1CH3 1J 1~ \
' `' ~T' ~'3 COC~3 XX~JT
,. ,_ ,.. , C~,~ C~ O ,~ \
~ 3C~I3 ~ .

~....~ .
' 3 i 3 ~
!l t \,, ~c~ c c~ c~
XXXVIII .
C~I C~ 0 ~ \
H3C\ ~C~ 1 3 . 1 3 ~ I O
O ~ -' `o~ ~t~
\-~ \C~ CH2CH3 25 X;~A1.~..
~I C CH C~3 CH 0 ~^\
3\~ .3 1 . 1 3 ll I O
0~ ~. Cx2CH3 ~ ~ ~C~

XL
H C C~ CH CH 0 ~ \
3\-~3~ 1 3 .~ 13 ~1 I n I O ~ T
\ ~ \CH C(CH3 )3 WO91/01~28 PCT/US90/040~2 XLI.
~A 3C, 13 1 1 Y I O ~

~ `~

2L~~.
o ~ ~
.~C~ 3-~ 3 '!

~ AA 3 ~-~_ A 3)2 XLIII.

3 3. ~ - 3 ~ i 0 ' `' ~ COc~2C~3 ~-~ ~CH
XLIV.
H3C~ ~CH3 ClH3, 1 U I~ O

i ,9~ ; COOCH3 XLV.
~3C, ,C~ C~3 C;i3 0 ~' I `0'~ -' `o' ~ ' \CH(C~I ) C~3 XLVI~
H3C~ ,~CH3 C~.3 '\ / \CH

WO 91/01128 PCI/US90/040~2 3 ~XLVII . --16--H3 C, ~CH3 7H3 . 1 8 I O

\ ~ \CH
XLVIII .
}~3C~ ~C~3 1CH3 CH3 o ~'\ ,C~`, Cri3 lo I O ~ o/ ~ ~
~, ~

XLIX .

15H3C~ ,~CX~t lCi3 ~ '3 0 ~ 50 ~ _H~

`.' `cx L.
I D ~ ^' `o~ ~ ~

\-/ \CH
25 LI.
3. 1 . 1 8 I O

\-~ \CH

LII .

3 . 3.~ 1 1 ll I O

\ ~ \CH

I` ~ '; ' ' ~`' .'`' The therapeutic agents of this invention may be administered alone or in combination with pharmaceutically-acceptable carriers, the proportion of which is determined by the solubility and chemical S nature of the compound, chosen route of administration and standard pharmaceutica' practice.
For example, they may be administered orally in the form of tablets or capsules con~aining s~ch excipients as starch, milk, sugar, certain of clay lO and so forth~ They may be administered erally in -~.~e form of solutions ~hich m2y contaln col~ir.~ o ~la~ori~ ag~nts~ '~'hen ap~ d to~ '. ^o:
treatmen~ o~ hotoagin~, ~he~ may be ~r_:.d~ in ~.e form of dus.in~ powders, aerosol sp ays~ oinL~ents~
lS aqueous compositions including solutions and suspensions, cream lotions and the like~ In this regard, any of the commonly employed ex.ending agents can be used depending on the nature of the product as is well-known in the art.
The physician will determine the dosage of the present theraputic agents which will be most suitable and it will vary with the form of administration and the particular compound chosen, and furthermore, it will var~ with the particular 25 patient under treatment. ~e will generally wish to initiate treatment with small dosages substantially less than the optimum dose of the compound and increase the dosage by small increments until the optimum ef_ect under the ci;cumstances is reached.
The polyenes which are formulated in moisturi- zing bases such as creams or ointments are usually provided in low concentrations. For example, Compound I may be used in co: entrations of about O.OOl percent to lO percent and preferably about O.Ol 35 percent to 5 percent by weight of the base.

J r j* ~ ~ Other non-toxic, dermatologically acceptable vehicles or carriers in which the polyenes are stable will Dc' e-~-ident to those of ordinary skill in the art. Tn ~er.Q-al ~ eem~ol 1' ent or lub ic_~ing vehicles, such as oleaginous substances, which help hydrate the skin -~ e -~eferred. As used herein, the term "emollien~" will be understood to refer to the nor.~ e~ ng ^'.~ ter o~ the composi,ion as a whele. ~rhat is, the nature o~ the vehicle and amount 10 of ~e~ srs ~ should be seiectQd so as to pro~ ?~ asin~ i~se r`or to-ical applica-tion. j'o.~ le ~ehicl~ hich dry Or ^~herwise harm ~h~ c~ ' a~à ~c~ ! s:~ould be avoided.
~ ~ '.--_ ?. "--. . '' ^~ 3 p r e--'~ - d i~. ' h_ ~ At er an_ :`n ;ubj 'C_3 ~' 'h very dry skin~ Examples oî suitable ointment bases are petrolatum, petrolatum plus volatile silicones, lanolin, and water in oil emulsions, such as Eucerin (Beiersdorf).
In warm weather and often for younger persons, oil in water emulsion (cream) bases, are preferred. Examples of suitable cream bases are Nivea Cream (Beiersdorf), cold cream (~SP), Purpose Cream 'Jch..son ~ Johnso-.~, hyd ophili; sintment (USP), and Lubriderm (Warner-Lambert).
The length of treatment of human skin can vary. Usually, there is little point in beginning the t,e^~tments of the present invention until young adult lire or, more typically, in middle age, when the ef~ects of aging begin to appear. The particular program of maintenance therapy according to the present invention will vary depending upon the individua. and conditions bein, treated. Generally, depending upon the age and state of the skin when treatments begin, it has been -ound tha once a day WO91/01128 PCT/US90/040~2 -19- C~ J
applications of polyenes for up to two months may be necessary to reduce and control the effects of aging which have already occurred. Once a stabilized skin controi ;l~3 been obtained, the frequency of applica-tion o~ t he ?olyen2s may be reduced, for example to ~WO `~ ?~ ~:im's a Wi e~;, an~ .-n SCm~Q cases only once a wee'; ior the res~ of ~he person's life. That iS, OïlCe ~he ~ing process hâs been controlled, a main~enance dose on the order of two appiications per 10 ~ee~ m~eral l ~t su..f icien~ to main~ain that state.
~ op cal compos.~icns o ~h s invention ca~ ir~d~ con~ abl2 suns..een agents~
An~ c^n~-~z~ion.al sunsc~ eQn ~.gQn~ can be utilized in ", ,1 ~ Q ~, Q ~ t~o-.s .;~.~ch c2n be '.lC_ n~ n, on.
~ hese topicai compvsi ions can contain any of the conventional excipien~s and addl ives commonly used in preparing topical compositions. Among the conventional additives or excipients which can be utilized in preparing these cosmetic compositions in accordance with this invention are preservatives, thickeners, perfumes and the like. In addition, the conventional antioxidants, such as butylated hydroxy-anisoles ~EA), ascorbyl palmitate, propyl gallate, citric aci butylated hydroxy toluene (~T), ethoxy-quin and the like can be incorporated into these compositions. These topical compositions can contain conventional acceptable ca; iers lor topical applica-tions which are generally utilized in these composi-tions~ These compositions may contain ~hickeningagents, humectants, emulsifying agents and viscosity stabilizers, such as those generally utilized. In addi~ion, these compositions can contain flavoring agents, colorants, and perfume which are conventional in preparing cosmetic compositions.

WO91/01~28 PCT/US90/04052 J~u~
This invention is further illustrated by the following examples, which are illustrative only.
Example A. Efficacv Te~
Compound II was tested for its effoc. on the differentiation of epidermis and do -i~ in hai ~ ? ~S
mice and directly compared to all-.rans re.inoic àcia.
In the first test used, polyene co~-npounds related to vitamin A, including all-tra~s re~inoic acid, are highly effective in reduc ns~ the si_~ o~`
horn-filled utricles in hairless rhine mouse sxi.~
The number of interutricular ep.d~Qr~al c~l~s le~ ss increases, concomitantl-, as the si20 of ~he ~ s.ls~
diminish. Increased num~ers of o?~do-mal CQ.
aro also prominent in human phc-'o__~ Si a ; _ _~
with all-trans retinoic acid.
~ airless rhino mice ~hrrhhrrh) from Temple University Skin and Cancer ~ospital were treated with 0.05 ml of Compound II, all-trans retinoic acid or the ethanol:acetone (1:1) vehicle on the dorsolateral skin once daily on five consecutive days, for four weeks. Mice were sacrificed by C02 on the third day after the last treatments. A 7/8"
punch biopsy of skin was removed and bisected in half. One of the halves was placed in 0.5 percent acetic acid overnight at 4C so that horizontal epidermal sheets could be removed from each biopsy.
The following day, epidermal sheets were removed from the dermis by peeling with a metal spatula. These sheets were fixed in formalin, dehydrated with ethanol, and kept in xylene. The other half of the biopsy was placed in 10 percent buffered formalin and processed into hematoxylin and eosln (~E~-stained vertical sections.

wosl/oll28 PCT/US90/04052 To assess utricle diameter, each epidermal sheet was placed on a glass slide in a rew drops of xylene. The diameter of 40 utricles was measured with an image analyzer~ The epidermal thickness of the ~&E-stained sections was measured on 15 inter-utricular areas of each section with ar, image analyzer~

WO9l/01128 PCT/US90/040~2 .
h 'J ~3 ~ 22--Table l ~cs~-Rel.~ed Activity o~ Compound II and All-Trans Retinoic Acid on ~hino ~ouse Skin Utricle Diameter ?ercPnt Con-Mean Percent c~n. ation,Diameter Reduction T~ `Q~?t~ !v llm~ * VS, V~ e Vehic;e i~8 ~ i~ 0 lO Cc~ 58 2 ~ i~ 55 ~Oûi /~ 46 0.0001 ll~ 2Q
All-Trans Retinoic Acid 0.1 54 ~ 8 61 0.01 62 ~ 9 55 0.001 70 ~ lO 49 0.0001 95 ~ 16 31 * 5 mice per group.
As seen in Ta~le 1, Compound I~ has marked activity over a wide concentration range, identical to all-trans retinoic acid.
The interutricular epidermal thickness results are shown in Table 2 (on skin samples from the same rhino mice that have their data in Table 1).

wos1/01128 PCT/US90/04052 Tab_e 2 Dose-Reiated Activity of Compound II and All-Trans Retinoic Acid on ~ o ~iouse ~;~in ~pidermal ThicXness ?e,cent Con- EpidermalPercent cen~ra ion, lhic:~ness Control Tre~m~n _ _w I v ~ vs ~ V~h i~l e P~ . , Compound ~ ~.5 $5~ + 8~9 236 ~ .3 ~ 234 v.v ~ 05 ; 186 ~Oi~i 35~ 152 All-Trans Retinoic Acid O~l 62.2 + 7.0 265 O.Ol 47.7 + l.3 203 0 . 001 37 . 6 ~ ? . 7160 0 . 0001 37 . 6 l 6 . 2160 *5 mice per group Compound II was as` effective as all-trans retinoic acid in increasing the interutricular epidermal thickness of rhino mice. This increase in epidermal thickness was due to an increase in the number of epidermal cell layers.
Polyene compounds are also evaluated for their effects on epidermàl differentiation when they are applied to a non-wrinkled strain of hairless mice (hrhr). These mice have fewer horn-filled utricles in their s~in compared to rhino mice. A variety of polyene compounds induce an increase in the number of epidermal cell layers when a compound is applied once to dorsal s~in.

WO 91/01128 PCr/US90/040a2 3 ~r) ~ 24--Hairless mice, obtained from Jackson Labs, had their dorsolateral skin treated twice, on Days 0 and 1, with O.OS ml of Compound II, all-trans retinoic acid, or ethanol vehicle. At the peak or the epidermal hyperplasia, on Day 4, the mic2 w~?re sacrificed by C0~ and a 7/8" punch biopsy rrom che treated skin was removed and placed into 10 pqrC?- ' buffered formalin. ~E-stained verticai secticns were prepared and the epidermal thic~ness in ~h~
interfollicular areas was measured wi~h aa ima~
analy~er.
The results are shown in Ta~le ~.

WO 91/01128 PCr/US90/040~i;2 --2 5~ , ,~ r r~
Table 3 ,~,, , ,_ ,j .~,!
Epidermal Hyperplasis in Hairless Mouse Skin Induced by Compound II and All-Trans Retinoic Acid Percent Con- EpidermalPerc~nt centration, Thickness Control Treatm~n~_ w/v _ ~ S.D.* V~ V~h;cl~
Vehicle 22.2 ~ 3.4 lO0 Compound II 0.1 56.9 ~ 2~6 2~5 0.01 53.4 ~ 3.0 23 Q.001 37.6 _ 6.1 16 All-Trans Retinoic Acid 0.1 4~ 3 - 6~ 7 ~^1 0.01 38.9 ~ 3.~ 17$
0.001 33.3 ' 5.3 149 * 5 mice per group.
At all three concentrations, Compound II
caused the same degree of epidermal hyperplasia as the three e~uivalent concentrations of all-trans retinoic acid.
All-trans retinoic acid is known to affect the differentiation of cells in the dermis of hair-less mouse skin, most effectively in the skin of mice that have been damaged by UV radiation. The forma-tion of a new zone of connective tissue is acceler-ated in photoaged hairless mouse skin by topical treatment with all-trans retinoic acid. This is due to an increased number of fibroblasts and an increase in their metabolic activity. As a result, new collagen bundles and elastic fibers are formed, which leads to a compression of the old, abnormal elastic fibers by the nèwly created dermal matrix.

,, j J~ -26-Female hairless mice (Skh-~Rl), six to eight weeks old, obtained from Temple University Skin and Cancer ~ospital, had their dorsal skin irradiated with ~ iol~ raaia~ion on ~onday, Wednes-day and ~ y each weeX for ten weeks, using a bankof o~ ~c-s~ ?~ d l~ cm above the ~as'; cf the mico Du ing ~he ~irst three weel;s, ~';e ~ .;a~ic/a dose pe~ wa~ ?rogressively increased -^.n one ~ nimal ?ry.hemal dose (~ED) to i~our `I3 s ~he ---iE~ dose ?er say was ~hen con-`;' '~e ~ . cf ~ vea-`iS~ ir~adi ation was stop~ed i~1i s.a- ri~C ~ ivee`i e.~ evQr~, tr~tment with a ~ 05 ~l ~t ~*~?n~ hi~l~, ?.~ ?~-S ~ ?.oic acld, o C~ nc~
dailY -o; ` ~-e consecu~i-ve days ror ten weeks~
.~' the end of treatme..-, mice were sacri-ficed by cervical dislocation and dorsal skin was removed and placed in lO percent buffered formalin.
Parafin-embedded sections were cut at lO ~m thickness and stained with Luna's aldehyde fuchsin for elastic fibers~ The dermal repair zone was measured microscopically and is defined as the area from the epidermal-dermal border to the top of the compressed elas.otic material The results in Table 4 show that Compound II
was as effective 2S all-trans retinoic acid in causin~ increased repair cf the connective tissue zone.

WO91/01128 PCT/US90/040~2 Table 4 !~'`';(; ~, The Depth of the Dermal Repair Zone Induced in Photoaged ~airless Mouse Skin ~y R~ resentati~ Compounds of Percent ~erm~l Repair Control o~e ne~th, ~m~`~ Vs~--ve~icle ~ a L ~ _ lO Untr~ 6.9 109 Vehicl~ .0 + ~l.4 lO0 Compoun~ O.l percent Q8 ~ + l~.~ 224 All-~.an~ inoie Acid, 0.05 percent llO.6 + 21.8 251-277 * 12 mice per group.
Part II

Compound XIII, O.l percent lOl.3 291 25 Compound IX, O~l percent 98.4 282 Compound V, O.l percent 90~72 260 All-T~ans Retinoic Acid, O~Ol percent 96.44 277 Vehicle 3~ 79 Part I,lI
Compound X, O.l percent 82.33 223 Compour.d XII, O.l percent 77.7~ 210 Compound X~;, O.l percent ~4 . D8 229 35 All-Trans Retinoic Acid, O.l pc~ca~t 7.50 264 Vehicle 36.88 WO 91/01128 PCI`/US90/040~2 r~ 2 8--.~v~

Compound XXII 93.71 414 5 All Trans-Retinoic Acid, 96.16 42j 0.1 percent Vehicle 22 . 6 0.1 percent B~ ToxicitY Tests A rabbit model of s};in i.r~a.isn ~va~
to assess the dermati~is produced bv- ~r ~3'.~en~
Compound II and all-trans relinoic acid Tho r~
is ccmmonly used as a skin irri~a~icn me~el predicting the potential local irrita~i~n of topically applied materials New Zealand albino rabbi~s~ rrem ~ec;iens Farms, Sanborn, N~Y., were clipped closely at _our sites on the back with an electric hair clipper to give 4 cm X 4 cm square sites. Each rabbit received 0.2 ml of Compound II and all-trans retinoic acid, once daily for fourteen consecutive days. Each day, the degree of erythema, scaling and edema was assessed visually by using the Draize O to 4 grading method. The results were expressed as average daily Draize score, which was derived by takir.g the cumula-tive score over fourteen days, for each parameter, and dividing by fourteen.
Table 5 shows a comparison of tnree doses of Compound II (0.1, 0.01 and O.OQl percent) to 0.01 percent all-trans retinoic acid.

WO91/Otl28 PCT/US90/040~2 --2 9~ ~ r .
Table 5 Mean Daily Draize Score Avera~ed Over 14 Days ' S.D~*
Treatment Global 5 (.percent) _Ery~hema Scaling ~ m~
Part.I
All-Trans Retinoic Acid,`
10 0.01 1~36 ~ 0.53 1.3 ~ 0.70 0~3 + 0~2 Compound ;T, 0.1 1.,1 _ 0.54 0.3 _ 0.57 O. . _ 0.3 0.01 0.ô3 + 0.57 G.~7 + 0.4S0. , O 5 0.001 0.52 ~ 0.33 0.1, + 0.19 0 + O
~10 rabbits.
Part_II
All-Trans Retinoic Acid, 0.1 6.6 Compound II 1.65 Compound XIX, 2.5 0.1 Compound XX, 4 5 0.1 30 Compound XXV, 3 3 0.1 Compound XXVI 3.3 0.1 n~ J Compound XXVII, 6.6 0.1 Compound ~ rv~ 6.6 O.l Compound ~ , 7 3 O . i ~ 0 ~ s ~
d~ ee o irri~ation caused bv Compol1~d ~1 l?t~ ? ~ ?~ ~.S ~ olc acid, is ~^~ s~ica'l~- lotve- ~an ail-t~ans retinoic ~ C ~ A ~ ~ - ? ~'.' ~ d ~ c~u~ ~f th~
15 ''2;`-~' ' ;V ~ssocia;ed '~ibli '`ile ~de~a ~cores, there were no sta~istically signi-icant differences between any t~eat.~ents, even though Compound II had numer-ically lower edema scores.
For the purposes of this invention, Global Irritation score is defined as the sum of the erythema, edema and scaling scores.
The toxicity induced by systemically administered Compound II was evaluated in albino mice. Vitamin A-related polyene compounds cause multipie signs o- toxicity, referred to as the hypervitaminosis A syndrome, characterized by loss of body weight, skin scaling, hair loss and bone ractures.
Albino CD-lTM mice, from Charles River Laboratories, Wilmington, Ma~, were administered Compound II and all-trans retinoic acid by intra-peritoneal injection in peanut oil, at 8 ml/kg, once daily $or ive consecutive days, for two weeks.
Mice were graded daily during treatment for signs of hypervitaminosis A by the method of Bollag.

wosl/0~128 PCT/US90~04052 -31- ~ ~., 5; ~ ~i An animal is defined as having hypervitaminosis A
when addition of the grades for loss of body weight, skin scaling, hair loss and bone fractures totals at least ~hr~e The results are shown in Table 6. Compound II a 3~ ihich ;as twico the hi_hest dose of all-..a..s .e'inoic acid, did not cause hyper-vi~ami~o~ ' 3 .~ ~ Iil con~rd3 L ~ at 100 mgl~g, all-trans retinoie acid was so ~o~ic tha. all ~n of ten mice showec,~ ?~;;itamino3is .~ and ~hree ~ice treated with t~..s ~ d ~c;~e~ 7 dnd '0 W09lt01~28 PCT/US90/04052 ~.~5~ 32-Table 6 Assessment of the Effects of All-Trans Retinoic Acid and Compound II on ~ypervitaminosis A Signs in CD-lTM ~ic~
Mean ~ypervitaminosis A
Treatment _ Çra~_+_S_P
All-Trans Retinoic Acid 100 mg/kg 5.
50 mg/kg 2.3 1 2.
1010 mg/kg O = 3 Peanut Oil Vehicle O
Untreated O _ O

Compound II
15200 mg/~
50 mg/kg ~ ~ v 20 mg/kg O ~ O
Peanut Oil Vehicle O + O
Untreated O + O
* Retinoid-treated groups had 10 mice. Untreated and vehicle-treated groups had 5 mice. Each retinoid was evaluated in separate studies.
** Based on 7 mice. Three mice died between Days 7 and 10.
In comparative treatment with 13-cis retinoic acid it was found that Compound II was approximately twice as effective as 13-cis retinoic acid and also caused less dermal irritation.
The invention has been described in detail with particular reference to preferred embodiments thereof, but it will be understood that variations and modifications can be effected wi~hin the spirit and scope of the invention.

Claims

What is claimed is:

1. A method of repairing sun-damaged human skin comprising topically administering a compound having the structure:

wherein R1, R2, R3, R4 and R5 are independently selected from the group consisting of H, Cl, OR6, straight or branched alkyl of 1 to 10 carbon atoms, NO2, COOR6, CN, OR6, NR6R7, NR6C(=S)NR7R8, NR6COR7, SO2NR6R7, CH(CH3)COOH, CONR6R7, COR6, OCONR6R7, NR6COONR7, R9OR6, NR6SO2R7, Si(CH3)3, and NR6CONR7R8, R3 together with R4 forms a benzo ring or taken together with R2 forms a benzo or tetrahydrobenzo ring or together with R2 and R1 forms a:

moiety or together with R2 forms a moiety or R2 together with R1 forms a benzo ring or R2 together with R3 forms a or or or moiety, or R1 is independently selected from the group consisting of moiety, R6, R7 and R8 are independently selected from the group consisting of straight or branched alkyl containing from 1 to 10 carbon atoms, aryl containing from 6 to 10 carbon atoms and hydrogen, and R9 is alkylene of 1 to 6 carbon atoms, and iron carbonyl complexes thereof, to an area of the skin in an amount sufficient to repair the effects of elastosis in the skin.

2. The method of claim 1 wherein R2 and R3 are independently selected from the group consisting of NR6COR7, CONR6R7, SO2NR6R7, OCONR6R7, NR6COOR7, NR6CONR7R8, NR6SO2R7 and NR6C(=S)NR7R8.

3. A method according to claim 1 wherein R3 is NHCOCH3 and R1, R2 and R4 are H.

4. A method according to claim 1 wherein the compound has the structure:

5. A method according to claim 1 wherein the compound is selected from the group consisting of:

, , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

6. A method according to claim 1 wherein said skin is human facial skin.

7. A method according to claim 1 wherein said compound is applied in an emollient vehicle.

8. A method according to claim 1 wherein the compound is applied with a dermatologically acceptable carrier.

9. The method of claim 1 wherein the compound comprises about 0.001 to about 10 percent by weight of the mixture applied.

10. The method of claim 1 wherein the compound comprises about 0.01 to about 5 percent by weight of the mixture applied.