Note: Descriptions are shown in the official language in which they were submitted.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
METHOD OF TREATING HAIR LOSS
FIELD OF THE INVENTION
The present invention relates to methods for treating hair loss in mammals,
including
arresting and / or reversing hair loss and promoting hair growth.
BACKGROUND OF THE INVENTION
Hair loss is a common problem which occurs, for example, through natural
processes or
is often chemically promoted through the use of certain therapeutic drugs
designed to alleviate
conditions such as cancer. Often such hair loss is accompanied by lack of hair
regrowth which
causes partial or full baldness.
As is well-known in the art, hair growth occurs by a cycle of activity which
involves
alternating periods of growth and rest. This cycle is often divided into three
main stages which
are known as anagen, catagen, and telogen. Anagen is the growth phase of the
cycle and may be
characterized by penetration of the hair follicle deep into the dermis with
rapid proliferation of
cells which are differentiating to form hair. The next phase is catagen, which
is a transitional
stage marked by the cessation of cell division, and during which the hair
follicle regresses
through the dermis and hair growth is ceased. The next phase, telogen, is
often characterized as
the resting stage during which the regressed follicle contains a germ with
tightly packed dermal
papilla cells. At telogen, the initiation of a new anagen phase is caused by
rapid cell
proliferation in the germ, expansion of the dermal papilla, and elaboration of
basement
membrane components. Wherein hair growth ceases, most of the hair follicles
reside in telogen
and anagen is not engaged, thus causing the onset of full or partial baldness.
There have been many attempts in the literature to invoke the regrowth of hair
by, for
example, the promotion or prolongation of anagen. Currently, there are two
drugs approved by
the United States Food and Drug Administration for the treatment of male
pattern baldness:
topical minoxidil (marketed as Rogaine by Pharmacia & Upjohn), and oral
fmasteride
(marketed as Propecia by Merck & Co., Inc.). For several reasons, however,
including safety
concerns and / or lack of efficacy, the search for efficacious hair growth
inducers is ongoing.
Interestingly, it is known that the thyroid hormone known as thyroxine ("T4")
converts
to thyronine ("T3") in human skin by deiodinase I, a selenoprotein. Selenium
deficiency causes
a decrease in T3 levels due to a decrease in deiodinase I activity; this
reduction in T3 levels is
strongly associated with hair loss. Consistent with this observation, hair
growth is a reported
side effect of administration of T4. See, e.~., Berman, "Peripheral Effects of
L-Thyroxine on
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
2
Hair Growth and Coloration in Cattle", Journal of Endocrinology, Vol. 20, pp.
282 - 292 (1960);
and Gunaratnam, "The Effects of Thyroxine on Hair Growth in the Dog", J. Small
Anim. Pract.,
Vol. 27, pp. 17 - 29 (1986). Furthermore, T3 and T4 have been the subject of
several patent
publications relating to treatment of hair loss. See, e.g., Fischer et al., DE
1,617,477, published
January 8, 1970; Mortimer, GB 2,138,286, published October 24, 1984; and
Lindenbaum, WO
96/25943, assigned to Life Medical Sciences, Inc., published August 29, 1996.
Unfortunately, however, administration of T3 and / or T4 to treat hair loss is
not
practicable because these thyroid hormones are also known to induce
significant cardiotoxicity.
See, e.~., Walker et al., U.S. Patent No. 5,284,971, assigned to Syntex,
issued February 8, 1994
and Emmett et al., U.S. Patent No. 5,061,798, assigned to Smith Kline & French
Laboratories,
issued October 29, 1991. Surprisingly, however, the present inventors have
discovered
compounds which promote hair growth without inducing cardiotoxicity.
Consistent with this
discovery, but without intending to be limited by theory, the present
inventors have surprisingly
discovered that the compounds useful in the present invention interact
strongly with hair-
selective thyroid hormone receptors but interact less strongly, or not at all,
with heart-selective
hormone receptors. These unique properties are, of course, not shared with T3
and / or T4.
Accordingly, the compounds described for use in the methods and compositions
herein are
cardiac-sparing compounds useful for treating hair loss, including arresting
and / or reversing
hair loss and promoting hair growth.
SUMMARY OF THE INVENTION
The present invention relates to methods for treating hair loss comprising
administering
a compound which has been found by the present inventors to be particularly
useful for treating
hair loss in mammals, including arresting and / or reversing hair loss and
promoting hair growth.
The compounds utilized in the present method have the structure:
Rt a R2
R4 \ X /
O / Ri \
I
R3 R5 Ri9
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
3
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides,
esters, and imides
thereof, wherein R,, R2, R3, R4, R5, Rb, R18, R,9, and X are defined herein.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to methods of using compounds and compositions
which
are particularly useful for treating hair loss in mammals, including arresting
and / or reversing
hair loss and promoting hair growth.
In addition to discovering that the present compounds are useful for treating
hair loss,
the present inventors have also surprisingly discovered that the preferred
compounds are
cardiac-sparing. The preferred compounds useful in the method of the present
invention are
therefore, as defined herein below, cardiac-sparing.
Publications and patents are referred to throughout this disclosure. All
references cited
herein are hereby incorporated by reference.
All percentages, ratios, and proportions used herein are by weight unless
otherwise
specified.
In the description of the invention various embodiments and/or individual
features are
disclosed. As will be apparent to the ordinarily skilled practitioner all
combinations of such
embodiments and features are possible and can result in preferred executions
of the invention.
As used herein, wherein any variable, moiety, group, or the like occurs more
than one
time in any variable or structure, its definition at each occurrence is
independent of its definition
at every other occurrence.
Definition and UsagLof Terms
The following is a list of definitions for terms used herein:
As used herein, "alkanoyl" is an alkyl substituted with oxo (C=O).
As used herein, "alkoxy" is an oxygen radical having an alkyl, alkenyl, or
alkynyl,
preferably an alkyl or alkenyl, and most preferably an alkyl substituent.
Examples of alkoxy
radicals include -O-alkyl and -O-alkenyl. An alkoxy radical may be substituted
or unsubstituted.
As used herein, "alkyl" is an unsubstituted or substituted saturated
hydrocarbon chain
radical having from 1 to about 15 carbon atoms; preferably from 1 to about 10
carbon atoms;
more preferably from 1 to about 6 carbon atoms; and most preferably from 1 to
about 4 carbon
atoms. Preferred alkyls include, for example, methyl, ethyl, propyl, iso-
propyl, and butyl.
As used herein, "aryl" is an aromatic ring radical which is either carbocyclic
or
heterocyclic. Preferred aryl groups include, for example, phenyl, benzyl,
tolyl, xylyl, cumenyl,
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
4
napthyl, biphenyl, thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl,
pyrimidinyl,
quinolinyl, triazolyl, tetrazolyl, benzothiazolyl, benzofuryl, indolyl,
indenyl, azulenyl, fluorenyl,
anthracenyl, oxazolyl, isoxazolyl, isotriazolyl, imidazolyl, pyraxolyl,
oxadiazolyl, indolizinyl,
indolyl, isoindolyl, purinyl, quinolizinyl, quinolinyl, isoquinolinyl,
cinnolinyl, and the like.
Aryls may be substituted or unsubstituted.
As used herein, "arylalkyl" is an alkyl radical substituted with an aryl group
or an aryl
radical substituted with an alkyl group. Preferred arylalkyl groups include
benzyl, phenylethyl,
and phenylpropyl. Arylalkyls may be substituted or unsubstituted.
As used herein, "biohydrolyzable amides" are amides of the compounds used in
the
present invention which do not interfere with the activity of the compound, or
that are readily
converted in vivo by a mammalian subject to yield an active compound.
As used herein, "biohydrolyzable esters" are esters of the compounds used in
the present
invention which do not interfere with the activity of the compound, or that
are readily converted
in vivo by a mammalian subject to yield an active compound.
As used herein, "biohydrolyzable imides" are imides of the compounds used in
the
present invention which do not interfere with the activity of the compound, or
that are readily
converted in vivo by a mammalian subject to yield an active compound.
As used herein, "cycloalkyl" is a saturated carbocyclic or heterocyclic ring
radical.
Preferred cycloalkyl groups include, for example, cyclobutyl, cyclopentyl, and
cyclohexyl.
Cycloalkyls may be substituted or unsubstituted.
As used herein, preferred "halogens" (or "halos" or the like) are bromine,
chlorine,
iodine, and fluorine, more preferably, bromine, chlorine, and iodine, even
more preferably
bromine and chlorine, and most preferably chlorine.
As used herein, "pharmaceutically acceptable" means suitable for use in a
human or
other mammal.
As used herein, "safe and effective amount of a compound" (or composition, or
the like)
means an amount that is effective to exhibit biological activity, preferably
wherein the biological
activity is arresting and / or reversing hair loss or promoting hair growth,
at the sites) of activity
in a mammalian subject, without undue adverse side effects (such as toxicity,
irritation, or
allergic response), commensurate with a reasonable benefit / risk ratio when
used in the manner
of this invention.
As used herein "salt" is a cationic salt formed at any acidic (e.g., carboxyl)
group, or an
anionic salt formed at any basic (e.g., amino) group. Many such salts are
known in the art.
Preferred cationic salts include the alkali metal salts (such as, for example,
sodium and
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
potassium), alkaline earth metal salts (such as, for example, magnesium and
calcium), and
organic salts. Preferred anionic salts include the halides (such as, for
example, chloride salts).
Such acceptable salts must, when administered, be appropriate for mammalian
use.
Methods of the Present Invention
The present invention relates to methods of treating hair loss comprising
administering a
composition comprising a compound having the structure:
Ri s R2
R4 \ X /
O / RI \ RS
I
R3 R5 R19
and pharmaceutically acceptable salts, hydrates, and biohydrolyzable amides,
esters, and imides
thereof, wherein:
(a) X is selected from oxygen, sulfur, and CHz;
(b) R, and Rz are each, independently, selected from hydrogen, halogen,
hydroxy,
trifluoromethyl, C, - C6 alkyl, C1 - C6 alkenyl, and C, - C4 alkoxy;
(c) R3 is selected from hydrogen, alkyl, aryl, and arylalkyl;
(d) R4 is selected from R4 is selected from the group consisting of hydrogen,
halogen, C,
- C4 alkyl, C, - C4 alkenyl, hydroxy, and C~ - C4 alkoxy;
(e) RS is selected from hydrogen, halogen, hydroxy, alkyl, C, - C4 alkenyl, C~
- C4
alkoxy, cycloalkyl, aryl, and arylalkyl;
(f) R6 is selected from alkyl, aryl, -CHZCH(NHz)COZH, -CHZCH(OH)COzH, -
CHZCR~R8NR9Rlo, -YC(O)R~1, -(CHz)"OH, -CHzC(R~z)(NHz)COzRIS, -
CHZCH(COOH)(NH)C(O)(CHz)mC(O)NH(CHz)nRzo, arid -CHZCRIZRIZC(O)OR13;
(g) R~ is selected from hydrogen and C, - C4 alkyl;
(h) R8 is selected from hydrogen and -C(O)R";
(i) R9 and Rlo are each, independently, selected from hydrogen, C, - C4 alkyl,
and C~
C4 alkanoyl;
(j) Y is selected from bond and C, - C4 alkyl;
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
6
(k) R" is selected from hydroxy, C, - C4 alkoxy, -NR,4R,5, C, - C4 alkyl, C, -
C4 alkenyl,
and C, - C4 alkynyl;
(1) R,~ is C, - C6 alkyl;
(m) R,3 is selected from C, - C~; alkyl, cycloalkyl, and arylalkyl;
(n) R,4 is selected from hydrogen and C, - C4 alkyl;
(o) R,5 is selected from hydrogen, C, - C4 alkyl, and C, - C4 alkanoyl;
(p) R,8 and R,9 are each, independently, selected from hydrogen, C, - C6
alkyl, C, - C6
alkenyl, hydroxy, and halogen;
(q) m is an integer from 2 to 4;
(r) n is an integer from 1 to 4; and
(s) RZO is an unsaturated, unsubstituted five- or six-membered monocyclic
heterocycle
containing from one to two nitrogen atoms as the only heteroatoms.
The X Moiety
X is selected from oxygen (-O-), sulfur (-S-), and -CHZ-. Preferably, X is
selected from
oxygen and -CHZ-. Most preferably X is oxygen.
The R, and R~ Moieties
R, and RZ are each, independently, selected from hydrogen, halogen, hydroxy,
trifluoromethyl, C, - C6 alkyl, C, - C6 alkenyl, and C1 - C4 alkoxy.
Preferably, R, and RZ are
each, independently, selected from hydrogen, halogen, trifluoromethyl, and C, -
C6 alkyl. More
preferably, R, and RZ are each, independently, selected from halogen,
trifluoromethyl, and C, -
C6 alkyl. Most preferably, R, and RZ are each, independently, selected from
halogen and C, - C6
alkyl. Preferred halogens are chlorine and iodine.
The R~ Moie
R3 is selected from hydrogen, alkyl, aryl, and arylalkyl. Preferably, R3 is
selected from
hydrogen, alkyl, and arylalkyl. Preferred alkyls for R3 are methyl and ethyl.
A preferred
arylalkyl for R3 is benzyl.
The R4 Moie
R4 is selected from the group consisting of hydrogen, halogen, C, - C4 alkyl,
C, - C4
alkenyl, hydroxy, and C, - C4 alkoxy. Preferably, R4 is selected from hydrogen
and halogen
(most preferably iodine). Most preferably, R4 is hydrogen.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
7
The RS Moie
RS is selected from hydrogen, halogen, hydroxy, alkyl, C, - C4 alkenyl, C, -
C4 alkoxy,
cycloalkyl, aryl, and arylalkyl. Preferably, RS is selected from hydrogen,
halogen (particularly
iodine), alkyl, cycloalkyl, aryl, and arylalkyl. More preferably, RS is
selected from hydrogen,
alkyl, cycloalkyl, aryl, and arylalkyl. Even more preferably, RS is selected
from alkyl,
cycloalkyl, aryl, and arylalkyl. Most preferably, RS is selected from alkyl
and arylalkyl (a
preferred arylalkyl is benzyl).
The R,8 and RI9 Moieties
R,8 and R19 are each, independently, selected from hydrogen, C, - C6 alkyl, C~
- C6
alkenyl, hydroxy, and halogen. Preferably, R,8 and R,9 are each,
independently, selected from
hydrogen and C, - C6 alkyl. Most preferably, Rl8 and R,9 are each hydrogen.
The R6 Moie
R6 is selected from alkyl, aryl, -CHzCH(NHz)COZH, -CHzCH(OH)COZH, -
CHzCR~R8NR9R~o, -YC(O)Rl,, -(CHz)n OH, -CHzC(R,z)(NHz)COzR~s, -
CHZCH(COON)(NH)C(O)(CHz)mC(O)NH(CHz)nRzo, and -CHZCR,zRIZC(O)OR,3. Preferably,
R6 is selected from -CHzCH(NHz)COZH, -CHZCH(OH)COZH, -CHZCR~R8NR9Rlo, -
YC(O)R,1, -
CHZC(R,z)(NHz)COZR,3, -CHzCH(COON)(NH)C(O)(CHz)",C(O)NH(CHz)"Rzo, arid -
CHZCR,zRIZC(O)OR~3.
R~ is a substituent on -CHZCR~R8NR9Rlo. R~ is selected from hydrogen and C~ -
C4
alkyl.
R8 is a substituent on -CHZCR~RgNR9R~o. R8 is selected from hydrogen and -
C(O)R~1.
Preferably R$ is hydrogen.
R9 and Rlo are substituents on -CHZCR~R8NR9R~o and CHZCR~RgNR9R,o. R9 and R,o
are
each, independently, selected from hydrogen, C, - C4 alkyl, and C1 - C4
alkanoyl (oxo substituted
alkyl). Preferably, R9 and R,o are each, independently, selected from hydrogen
and CI - C4 alkyl.
Y is a substituent on -YC(O)R". Y is selected from a bond and C, - C4 alkyl.
Wherein
Y is bond, the C(O)Rl, of -YC(O)R" is directly covalently linked through a
single bond to the
respective phenyl ring of the compound.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
8
R" is selected from hydroxy, C, - C4 alkoxy, -NR,4R,5, C, - CQ alkyl, C, - C4
alkenyl,
and C, - C4 alkynyl. Preferably, R" is selected from hydroxy, C, - C4 alkoxy,
and -NR,4R,s.
Most preferably, R" is selected from hydroxy and C, - C4 alkoxy.
R,z is a substituent on -CHzC(R,z)(NHz)COzR,3 and -CHzCR,zR,zC(O)OR,3. R,z is
C, -
C6 alkyl.
R,3 is a substituent on -CHZCR,zR,zC(O)OR,3 and -CHZC(R,z)(NHz)COZR,3. R,3 is
selected from C, - C6 alkyl, cycloalkyl, and arylalkyl. Preferably, R,3 is C, -
C6 alkyl.
R,4 and R,5 are each (optionally) substituents on R". R,4 is selected from
hydrogen and
C, - C4 alkyl, preferably methyl or ethyl. R,5 is selected from hydrogen, C, -
C4 alkyl, and C, -
C4 alkanoyl (oxo (C=O) substituted alkyl). Preferably, R,; is selected from
hydrogen and C, - C4
alkyl.
The integers m and n are of -CHzCH(COON)(NH)C(O)(CHz)mC(O)NH(CHz)"Rzo. The
integer m is from 2 to 4, preferably 2. The integer n is from 1 to 4,
preferably 1. Rzo is also of -
CHzCH(COOH)(NH)C(O)(CHz)mC(O)NH(CHz)"Rzo. Rio is an unsaturated, unsubstituted
five-
or six-membered monocyclic heterocycle containing from one to two, preferably
one, nitrogen
atoms as the only heteroatoms. Non-limiting examples of preferred heterocycles
are pyrrolyl,
pyrazolyl, imidazolyl, pyridyl, pyridazinyl, pyrimidinyl, and pyrazinyl, more
preferably
imidazolyl, most preferably s-imidazolyl.
Preferred compounds useful in the method of the present invention are reported
in
Yokoyama et al., "Synthesis and Structure - Activity Relationships of Oxamic
Acid and Acetic
Acid Derivatives Related to L-Thyronine", Journal of Medicinal Chemistry, Vol.
38, pp. 695 -
707 (1995). These compounds are further described in Table 1 below:
Table 1
Rz
Rs \ O / NH2
HO / Rl \ C02H
R~ Rz Rs
methyl methyl iodine
bromine bromine iso-propyl
methyl methyl iso-propyl
hydrogenhydrogeniodine
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
9
Other preferred compounds useful in the methods of the present invention are
those
described in Li et al., WO 99/00353, assigned to Karo Bio, published January
7, 1999.
Particularly preferred among these compounds have the structure:
R2
Rs \ O / O
I/ \I
HO R~ Y R1 ~
wherein RS is selected from C, - C6 alkyl and C3 - C7 cycloalkyl; R, and R~
are each,
independently, selected from hydrogen, halogen, and C, - C6 alkyl, wherein at
least one of R,
and RZ is not hydrogen; and R" is selected from the group consisting of
hydroxy and C, - C4
alkoxy. Preferred compounds of this structure are presented below in Table 2:
Table 2
Cl
O
\ /
I I
HO / C1 \ CH2COOH
Br
O
\ /
I I
HO / \ CH2COOH
Other preferred compounds used in the methods of the present invention are
disclosed in
Kun et al., WO 97/46228, assigned to Octamer, Inc., published December 11,
1997. Particularly
preferred among these have the structure:
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
RI8 R2
R4 ~ X
~ I O
O ~ \ RI v
R3 R5 \Rl~ OCH3
wherein R3 is selected from methyl and ethyl; R4 is selected from hydrogen and
iodine; RS is
selected from hydrogen, iodine, and alkyl; R,8 and R,9 are each,
independently, selected from
hydrogen and C~ - C6 alkyl; X is selected from oxygen, sulfur, and CHZ; and R,
and RZ are each,
independently, selected from hydrogen, halogen, and C~ - C4 alkyl.
Other preferred compounds useful in the methods of the present invention are
those
described in Kun et al., WO 99/20263, assigned to Octamer, Inc., published
April 29, 1999.
Other preferred compounds useful in the methods of the present invention are
those
described in Feinber ee t al., U.S. Patent No. 4,711,855, assigned to Ciba
Corning Diagnostics
Corp., issued December 8, 1987. Particularly preferred among these compounds
have the
structure:
I
OH
I ~ O / O O ~CH~ RZo
NH'L
HO / I \ N CH2~--
H O
wherein m is an integer from 2 to 4; n is an integer from 1 to 4; and RZO is
an unsaturated,
unsubstituted five- or six-membered monocyclic heterocycle containing from one
to two
nitrogen atoms as the only heteroatoms. Examples of these heterocycles
include, but are not
limited to, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyridazinyl,
pyrimidinyl, and pyrazinyl, more
preferably imidazolyl, most preferably 5-imidazolyl.
Other preferred compounds useful in the methods of the present invention are
those
described in Wechter et al., U.S. Patent No. 3,449,419, assigned to The Upjohn
Co., issued June
10, 1969. Particularly preferred among these compounds have the structure:
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
11
R4 R~8 R2
O ~ / O ~ ~ YC(O)R1 ~
R3
R5 R~
R19
wherein R3 is selected from hydrogen and C, - C4 alkyl; R4 is selected from
hydrogen and iodine;
RS is selected from hydrogen, iodine, and C, - C4 alkyl; R,g and R,9 are each,
independently,
selected from hydrogen and C, - C4 alkyl; R, and RZ are each, independently,
halogen; Y is
selected from bond and C~ - C3 alkyl; R,~ is -NR14R~5; and R,4 and R~5 are
each, independently,
selected from hydrogen and C, - C4 alkyl.
Other preferred compounds useful in the methods of the present invention are
those
described in Kummer et al., U.S. Patent No. 3,930,017, issued December 30,
1975. Particularly
preferred among these compounds have the structure:
R2
Rlz Rlz
O
HO~/O~~ ~Ri3
O
I RI
wherein R4 is selected from hydrogen and iodine; R, and Rz are each,
independently, selected
from hydrogen and iodine; each RIZ is, independently, C, - C6 alkyl; and R,3
is selected from C, -
C6 alkyl, cycloalkyl, and arylalkyl.
Other preferred compounds useful in the methods of the present invention are
those
described in Ellis et al., U.S. Patent No. 4,766,121, assigned to Smith Kline
& French
Laboratories Ltd., issued August 23, 1988. Particularly preferred among these
compounds have
the structure:
R2
X
\ /
HO / R~ \ R6
Ri6 R~7
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
12
wherein X is selected from oxygen, sulfur, and CHI; R~ and R~ are each,
independently, selected
from hydrogen, halogen, and C, - C4 alkyl; R6 is selected from -CH~CR7R8NR9R,o
and -
YC(O)R"; R,6 is aryl; and R,~ is selected from hydrogen and C, - C4 alkyl.
Preferably, X is
oxygen; R" is hydrogen; R6 is -CHZCR,R8NR9R,o; Rg is -C(O)R"; R~, is hydroxy;
and / or R9 is
C, - C4 alkyl and R,o is hydrogen. Also preferably, R6 is -YC(O)R"; Y is C, -
C4 alkyl; and / or
R~ and RZ are each, independently, halogen. Particularly preferred groups for
R,6 are selected
from 4-hydroxyphenyl, 5-hydroxy-2-pyridyl, 6-oxo-3(1H)pyridyl, and 6-oxo-
3(1H)pyridazinyl.
Other preferred compounds useful in the methods of the present invention are
those
described in Leeson et al., "Selective Thyromimetics. Cardiac-Sparing Thyroid
Hormone
Analogues Containing 3'-Arylmethyl Subsituents", Journal of Medicinal
Chemistry, Vol. 32, pp.
320 - 336 (1989). Particularly preferred among these compounds are those shown
in Table 3
below:
Table 3
I
O
HO \ / O
OOH
I NHI
I
O
HO ~ / O
OOH
I NHI
CN
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
13
I
O
HO ~ / O
~OH
I NHI
N02
I
O
HO ~ / O
~OH
I NHI
HO
I
O
HO ~ / O
~OH
I NHI
C1
OH
I
O
HO ~ / O
OOH
I NHI
OH
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
I4
I
O
HO ~ / O
~OH
I NHI
N
I
O
HO ~ / O
~OH
I NHI
NH
O
I
O
HO ~ / O
I ~OH
I NH~
NH
O
I
O
HO ~ / O
~OH
I NHI
N
NH
O
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Br
O
HO \ / O
~OH
Br NHI
NH
O
O
HO ~ / O
~OH
NH
NH
O
Br
O
HO ~ / S
OOH
Br NHI
NH
O
Br
HO ~ / O
~OH
Br ,I0
NH
O
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
16
Br
HO ~ ~ O
Br ~ OH
O
NH
O
Br
O
HO ~ / O
l ~OH
Br NH2
N
NH
O
I
O
HO ~ / O
j ~OH
I NH2
OH
CA 02374262 2001-11-29
WO 00/72811 PCTNS00/05250
17
I
- ~ ~ O
HO ~ ~ O
OOH
I NHI
F
I
O
HO ~ / O
OOH
I NH~
NHZ
I
O
HO ~ ~ O
j OOH
I NH2
F
OH
I
O
HO ~ / O
~OH
I NHI
CH3
OH
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
18
I
O
HO ~ / O
~OH
I NH/
-N
I
O
HO ~ / O
~OH
I NHI
-N
OH
I
O
HO ~ / O
I ~OH
I NH~
N
O
I
O
HO ~ / O
OOH
I NHI
N
NH'~\
O
CA 02374262 2001-11-29
WO 00/72811 PCT/USOO105250
19
I
O
HO ~ / O
OOH
I NHI
N
O
C1
O
HO ~ / O
~OH
CI NHI
NH
O
I
O
HO ~ / S
OOH
I NHI
NH
O
I
O
HO ~ / O
OOH
I NH2
NH
O
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Br
HO \ / O ~ ~ O
Br OH
NH
O
I
O
HO ~ / O
j OOH
I NH2
NH
O
C1
O
HO ~ / O
OOH
Cl NHI
N~
1
NH
O
Analytical Methods
The present invention relates to methods of treating hair loss by
administering a
compound having a structure as described herein. Preferably, the compound
utilized in the
present invention will be cardiac-sparing. Compounds (test compounds) may be
tested for their
ability to induce anagen and their lack of cardiotoxicity (cardiac-sparing)
using the following
methods. Alternatively, other methods well-known in the art may be used (but
with the term
"cardiac-sparing" being defined according to the method disclosed herein
below).
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
21
Cardiotoxicity Assay:
The cardiotoxicity assay measures the potential of a test compound to
adversely affect
the cardiovascular system. As thyroid hormone (T3) damages the cardiovascular
system, the
heart enlarges. See, e.g., Gomberg-Maitland et al., "Thyroid hormone and
Cardiovascular
Disease", American Heart Journal, Vol. 135(2), pp. 187-196 (1998); Klein and
Ojamaa,
"Thyroid Hormone and the Cardiovascular System", Current Opinion in
Endocrinology and
Diabetes, Vol. 4, pp.341-346 (1997); and Klemperer et al., "Thyroid Hormone
Therapy and
Cardiovascular Disease", Progress in Cardiovascular Diseases, Vol. 37 (4), pp.
329-336 (1996).
This increases the weight of the heart relative to whole body weight. The
cardiotoxicity assay
herein below is used to test compounds for potentially adverse cardiac effects
by measuring their
effect on the heart-to-body weight ratio.
Two groups each of six male Sprague Dawley rats (Harlan Sprague Dawley, Inc.,
Indianapolis, IN) (each weighing from approximately 220 grams to 235 grams)
are utilized. The
first group is a vehicle control group and the second group is a test compound
group. The length
of the assay is 30 days, with treatment of vehicle or test compound in vehicle
daily for 28 of
those days as described below.
Prior to initiation of the assay, each rat is allowed to acclimate to standard
environmental conditions for 5 days. Each rat receives food (standard rat chow
diet) and water
ad libiturn 5 days prior to initiation of the assay as well as to termination
of the study.
The vehicle is 91:9 (v:v) propylene glycol:ethanol. The test compound is
prepared at a
concentration of 500 ~glmL in the vehicle.
Each rat is weighed on day 1 of the assay. Dosage calculations are then
performed: each
rat will be administered daily a dosing solution of vehicle or test compound
in vehicle
(depending on whether the rat is in the vehicle control group or the test
compound group,
respectively) at 500 ~L of dosing solution per kg of rat. For rats in the test
compound group,
this corresponds to a dose of 250 ~.g of test compound per kg of rat.
Day 2 is the first day of treatment with dosing solution for both groups. Body
weights
are taken for each rat on days 3, 5, 8, 10, 12, 15, 17, 19, 22, 24, 26, and 29
prior to dosing for
that day; for each rat, the dosing solutions are recalculated and administered
accordingly upon
change in body weight.
Treatment occurs once daily in the morning on days 2 through 29, inclusive,
for each rat
in each group. For each treatment, the dosing solution is administered
subcutaneously between
the shoulders of the rat such that the injection sites are rotated in this
area.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
22
On day 30 in the morning, the rats of each group are euthanized with COZ from
dry ice.
Each rat is immediately weighed for total body weight.
The hearts of each rat are then excised as follows. An incision is made to
expose the
abdominal cavity. The rib cage is carefully cut at the sternum with small
scissors, such that the
heart and lungs are exposed. With small scissors and forceps, the vessels
connected to the heart
are cut away from the heart. These vessels include the caudal vena cava, left
cranial vena cava
(pulmonary trunk), right cranial vena cava, thoracic aorta, right subclavian
artery, internal
thoracic artery and vein, and any other small attachments. The heart is then
immediately taken
out intact, including the left and right auricles and left and right
ventricles. Immediately
thereafter, any excess tissue is trimmed away, the heart is lightly blotted on
a paper towel until
no more blood is visibly left behind on the paper towel, and the heart is
weighed.
The heart weight is divided by the body weight after euthanization for each
rat to give
the heart/body ratio. The heart/body ratios for each rat in the vehicle
control group are added
together and divided by 6 (i.e., the total number of rats in the group) to
give RV (ratio for
vehicle control group). Similarly, the heart/body ratios for each rat in the
test compound group
are added together and divided by 6 to give RT (ratio for test compound
group).
The index C is then calculated by dividing RT by RV. As defined herein, where
C is
less than 1.3, the test compound is cardiac-sparing. Preferably, C is less
than 1.2, more
preferably less than 1.15, and most preferably less than 1.1. In accordance
with this method, T3
and T4 are not cardiac-sparing.
Telogen Conversion Assay:
The Telogen Conversion Assay measures the potential of a test compound to
convert
mice in the resting stage of the hair growth cycle ("telogen"), to the growth
stage of the hair
growth cycle ("anagen").
Without intending to be limited by theory, there are three principal phases of
the hair
growth cycle: anagen, catagen, and telogen. It is believed that there is a
longer telogen period in
C3H mice (Harlan Sprague Dawley, Inc., Indianapolis, IN) from approximately 40
days of age
until about 75 days of age, when hair growth is synchronized. It is believed
that after 75 days of
age, hair growth is no longer synchronized. Wherein about 40 day-old mice with
dark fur
(brown or black) are used in hair growth experiments, melanogenesis occurs
along with hair
(fur) growth wherein the topical application of hair growth inducers are
evaluated. The Telogen
Conversion Assay herein below is used to screen compounds for potential hair
growth by
measuring melanogenesis.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
23
Three groups of 44 day-old C3H mice are utilized: a vehicle control group, a
positive
control group, and a test compound group, wherein the test compound group is
administered a
compound used in the method of the present invention. The length of the assay
is at least 19
days with 15 treatment days (wherein the treatment days occur Mondays through
Fridays). Day
1 is the first day of treatment. Most studies will end on Day 19, but a few
may be carried out to
Day 24 if the melanogenesis response looks positive, but occurs slowly. A
typical study design
is shown in Table 4 below. Typical dosage concentrations are set forth in
Table 4, however the
skilled artisan will readily understand that such concentrations may be
modified.
Table 4
Group Animal CompoundConcentrationApplication Length of
# #
volume Study
1 1 - 10 Test 0.1% in 400 ~,L topical19 or 24
days
Compoundvehicle**
2 11 - Positive0.01 % in 400 ~,L topical19 or 24
20 days
Control vehicle**
(T3)
3 21 - Vehicle**N/A 400 pL topical19 or 24
30 days
**The vehicle is 60% ethanol, 20% propylene glycol, and 20% dimethyl
isosorbide
(commercially available from Sigma Chemical Co., St. Louis, MO).
The mice are treated topically Monday through Friday on their lower back (base
of tail
to the lower rib). A pipettor and tip are used to deliver 400 ~L to each
mouse's back. The 400
~L application is applied slowly while moving hair on the mouse to allow the
application to
reach the skin.
While each treatment is being applied to the mouse topically, a visual grade
of from 0 to
4 will be given to the skin color in the application area of each animal. As a
mouse converts
from telogen to anagen, its skin color will become more bluish-black. As
indicated in Table 5,
the grades 0 to 4 represent the following visual observations as the skin
progresses from white to
bluish-black.
Table 5
Visual Observation Grade
Whitish Skin Color 0
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
24
Skin is light gray (indication of initiation1
of anagen)
Appearance of Blue Spots
Blue Spots are aggregating to form one 3
large blue area
Skin is dark blue (almost black) with 4
color covering majority
of treatment area (indication of mouse
in full anagen)
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Method of Making
The compounds used in the methods of the present invention are prepared
according to
procedures which are well-known to those ordinarily skilled in the art. The
starting materials
used in preparing the compounds are known, made by known methods, or are
commercially
available as a starting material.
It is recognized that the ordinarily skilled artisan in the art of organic
chemistry can
readily carry out standard manipulations of organic compounds without further
direction.
Examples of such manipulations are discussed in standard texts such as J.
March, Advanced
Organic Chemistry, John Wiley & Sons (1992).
The ordinarily skilled artisan will readily appreciate that certain reactions
are best
carried out when other functionalities are masked or protected in the
compound, thus increasing
the yield of the reaction and / or avoiding any undesirable side reactions.
Often, the artisan
utilizes protecting groups to accomplish such increased yields or to avoid the
undesired
reactions. These reactions are found in the literature and are also well
within the scope of the
skilled artisan. Examples of many such manipulations can be found in, for
example, T. Greene,
Protecting Groups in Organic Synthesis, John Wiley & Sons (1981).
The compounds of the present invention may have one or more chiral centers. As
a
result, one may selectively prepare one optical isomer, including
diastereomers and enantiomers,
over another, for example by chiral starting materials, catalysts or solvents,
or may prepare both
stereoisomers or both optical isomers, including diastereomers and enantiomers
at once (a
racemic mixture). Since the compounds of the invention may exist as racemic
mixtures,
mixtures of optical isomers, including diastereomers and enantiomers, may be
separated using
known methods, such as through the use of, for example, chiral salts and
chiral chromatography.
In addition, it is recognized that one optical isomer, including a
diastereomer and
enantiomer, or a stereoisomer, may have favorable properties over the other.
Thus, when
disclosing and claiming the invention, when one racemic mixture is disclosed,
it is clearly
contemplated that both optical isomers, including diastereomers and
enantiomers, or
stereoisomers substantially free of the other are disclosed and claimed as
well.
The syntheses of the compounds useful in the present invention are described
in the art.
Accordingly, the ordinarily skilled artisan will be able to prepare the
compounds described
herein. For further guidance, the syntheses of various of the present
compounds are described
in, for example, Ellis et al., U.S. Patent No. 4,910,305, assigned to Smith
Kline & French
Laboratories, issued March 20, 1990; Ellis et al., U.S. Patent No. 4,826,876,
assigned to Smith
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
26
Kline & French Laboratories, issued May 2, 1989; Ellis et al., U.S. Patent No.
4,766,121
assigned to Smith Kline & French Laboratories, issued August 23, 1988; Emmett
et al., U.S.
Patent No. 5,061,798, assigned to Smith Kline & French Laboratories, issued
October 29, 1991;
Blank et al., U.S. Patent No. 3,149,153, assigned to Smith Kline & French
Laboratories, issued
September 15, 1964; Blank et al., U.S. Patent No. 3,287.396, assigned to Smith
Kline & French
Laboratories, issued November 22, 1966; Leeson et al., EP 0,188,351, assigned
to Smith Kline
& French Laboratories, published July 23, 1986; Steams, WO 90/07330, assigned
to The
Regents of the University of California, published July 12, 1990; Yu et al.,
U.S. Patent No.
4,363,815, issued December 14, 1982; Kummer et al., U.S. Patent No. 3,930,017,
issued
December 30, 1975; Kummer et al., U.S. Patent No. 4,110,470, issued August 29,
1978;
Wechter et al., U.S. Patent No. 3,449,419., assigned to The Upjohn Company,
issued June 10,
1969; Feinber~, U.S. Patent No. 4,711,855, assigned to Ciba Corning
Diagnostics Corp., issued
December 8, 1987; Kun et al., WO 97/46228, assigned to Octamer, Inc.,
published December 11,
1997; Li et al., WO 99/00353, assigned to Karo Bio, published January 7, 1999;
and Leeson et
al., "Selective Thyromimetics. Cardiac-Sparing Thyroid Hormone Analogues
Containing 3'-
Arylmethyl Substituents", Journal of Medicinal Chemistry, Vol. 32, pp. 320 -
336 (1989).
The following non-limiting examples provide even further guidance of making
the
compounds used in the present method.
Example 1
Br
_ O
HO ~ / O
OH
Br
To a suspension of bis(3-isopropyl-4-methoxyphenyl)iodonium tetrafluoroborate
(prepared by the method of Yokohama et al., "Synthesis and Structure -
Activity Relationships
of Oxamic Acid and Acetic Acid Derivatives Related to L-Thyronine", Journal of
Medicinal
Chemistry, Vol. 38, pp. 695 - 707 (1995)), and copper bronze (6.1 g) in
dichloromethane (150
mL), is added a solution of methyl 3,5-dibromo-4-hydroxybenzoate (15 g) and
triethylamine (5.4
g) in dichloromethane (100 mL) dropwise at room temperature. The mixture is
stirred overnight
and then filtered through celite. After concentration, the resulting residue
is passed through a
short silica gel column. The pure fractions are combined and concentrated to
dryness. The
residue is recrystallized from methanol to afford methyl 3,5-dlbromo -4-
(4'methoxy-3'-
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
27
isopropylphenoxy)-benzoate.
The above ester (6.5 g) is hydrolyzed by treatment with 1 M aqueous NaOH (60
mL)
and methanol (150 mL) to give 3,5-dibromo -4-(4'-methoxy-3'-
isopropylphenoxy)benzoic acid.
The benzoic acid (2 g) is demethylated with boron tribromide (1M, 26 mL) in
dichloromethane at 0 °C. The mixture is stirred overnight at room
temperature before quenching
with a water/ice mixture. The layers are separated and the water layer is
extracted with
dichloromethane. The combined organic extracts are dried, filtered and
concentrated. The
resulting residue is recrystallized to give 3,5-dibromo-4-(4-hydroxy-3-
isopropylphenoxy)benzoic
acid.
Example 2
Br
He O
~OH
Br //O
Bromine (35.2 g) is added dropwise to a suspension of methyl 4-hydroxy-
phenylacetate
(16.6 g) in water (500 mL). After 1 day of stirring, the mixture is
partitioned between water and
ethyl acetate. The organic layer is washed with aqueous sodium thiosulfate,
dried, filtered and
concentrated. The residue is recrystallized from methanol to give methyl 3,5-
dibromo-4-
hydroxyphenylacetate.
The above phenol (5 g) is coupled to bis(3-isopropyl-4-methoxyphenyl)iodonium
tetrafluoroborate (9.5 g) as described in Example 1. Purification by column
chromatography and
recrystallization from methanol affords methyl 3,5-dibromo-4-(4-methoxy-3-
isopropyl-
phenoxy)phenylacetate.
The above ester (2.4 g) is demethylated with boron tribromide. The crude
product is
recrystallized from dichloromethane and light petroleum ether to give 3,5-
dibromo-4-(4'-
hydroxy-3'-isopropylphenoxy)phenylacetic acid.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
28
Example 3
C1
_ O
HO ~ ~ O
OH
C1
Methyl 3,5-dichloro-4-hydroxybenzoate (10 g) is coupled with bis(3-isopropyl-4-
methoxyphenyl)iodonium tetrafluoroborate (35 g) using the method described in
Example 1.
Purification by column chromatography followed by recrystallization from
methanol affords
methyl 3,5-dichloro-4-(4-methoxy-3-isopropylphenoxy) benzoate.
The above methoxy compound ( 100 mg) is demethylated and hydrolyzed using the
method described in Example 1 to give 3,5-dichloro-4-(4-hydroxy-3-
isopropylphenoxy)benzoic
acid.
Example 4
Cl
HO ~ / O
OH
C1
Methyl 3,5-dichloro-4-(4-methoxy-3-isopropylphenoxy)benzoate (see Example 3)
(3.0
g) is treated with a 1M solution of diisobutyl aluminium hydride (DIBAL) in
tetrahydrofuran
(32.5 mL) at 0 °C and then warmed to room temperature and stirred
overnight. The reaction
mixture is poured into an ice-cold 1M HCl solution and extracted with ethyl
acetate three times.
The organic layer is washed (brine), dried, filtered and concentrated to
dryness to afford 3,5-
dichloro-4-(4-methoxy-3-isopropylphenoxy) benzylalcohol.
The above alcohol (200 mg) is demethylated using the method described in
Example I to
give 3,5 dichloro-4-(4-hydroxy-3-isopropylphenoxy) benzylalcohol.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
29
Example 5
I
O
O ~ ~ O O-
I
Methyl 3,5-diiodo-4-(4'-ethoxyphenoxy)benzoate is synthesized using the
general
methodology of Borrows et al., "The Synthesis of Thyroxine and Related
Substances. Part I.
The Preparation of Tyrosine and Some of its Derivatives, and a New Route to
Thyroxine",
Journal of the Chemical Society, Supp. Issue No. 1, S185 - 5190 (1949).
Example 6
I
_ O
O
OH
I
3,5-diiodo-4-(4'-methoxyphenoxy)benzoic acid is synthesized using the general
methodology of Borrows et al., "The Synthesis of Thyroxine and Related
Substances. Part I.
The Preparation of Tyrosine and Some of its Derivatives, and a New Route to
Thyroxine",
Journal of the Chemical Society, Supp. Issue No. l, 5185 - 5190 (1949).
Example 7
I I
HO ~ / O
~OH
I ~~O
3, 3', 5-triiodothyroacetic acid is commercially available from Sigma Chemical
Co., St.
Louis, MO.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Example 8
I I
HO ~ ~ O ~ \ O
I OH
3, 3', 5-triiodothyropropionic acid is commercially available from Sigma
Chemical Co., St.
Louis, MO.
Example 9
OH \O \O I
\ \ \ HO /
+ I
/ / 2 I \ CN
I+
9a 9b
OH
~O OH
\ \
\ \ \
/ I
/ I / I .-; O
O / O / OH
I \
I \ CN I \ CN 9e O
9c 9d
9a. 2-isopropyl anisole: Potassium hydroxide (5.6 g) is added to 13.4 mL
acetone followed by
2-isopropylphenol (13.6 g). After the potassium hydroxide is dissolved, methyl
iodide (14.2 g)
is added. The reaction is refluxed for about 16 hours. 150 mL of water is
added. This reaction
is extracted 3 times with 100 mL diethyl ether. The organic layer is extracted
twice with 100
mL 10% sodium hydroxide in water, once with 100 mL water, and once with 100 mL
saturated
ammonium chloride. After drying over magnesium sulfate, the organic solution
is dried over
MgS04, filtered, and concentrated under reduced pressure. The material is
fractionally distilled
under reduced pressure to afford 9a.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
31
9b. Bis(3-isopropyl-4-methoxyphenyl)iodonium Tetrafluoroborate: Acetic
anhydride (7
xnL,) is cooled to -15 °C in a dry ice/acetone bath. Fuming nitric acid
(5.4 mL) is added
dropwise. Iodine (2.5 g) is added in one piece followed by dropwise addition
of 4.7 mL
trifluoroacetic acid. After 20 minutes, the reaction is removed from the bath
and stirred at room
temperature for 30 minutes. After the iodine has dissolved, the reaction is
sparged to remove
nitrogen oxides and then concentrated under vacuum. The material is then taken
up in 15 mL
acetic anhydride and cooled to -10 °C. To this cooled solution is added
dropwise a solution of 2-
isopropyl anisole (9a; 7.43 g) in 35 mL acetic anhydride and 5 mL
trifluoroacetic acid. The
reaction is allowed to stand in a refrigerator for about 16 hours. After
allowing the reaction to
return to room temperature for 3 hours, the reaction is concentrated under
high vacuum. The
residue is taken up in 25 mL methanol, 25 mL 10% sodium bisulfate, and 188 mL
2M sodium
tetrafluoroborate. The mixture is stirred vigorously for 30 minutes and the
supernatant is
decanted off. To the residue is added 200 mL hexane and it is stirred for an
additional 30
minutes. At that time, the solid is collected, washed with hexane, and dried
under vacuum to
afford 9b.
9c. 2',6'-diiodo-3-isopropyl-4-methoxy-4'-nitrilediphenyl ether: Bis(3-
isopropyl-4-
methoxyphenyl)iodonium tetrafluoroborate (9b; 1 g) is weighed is taken up in 3
mL
dichloromethane and 0.17 g copper bronze is added. The mixture is cooled in an
ice water bath.
A solution of 2,6-diiodo-4-nitrilephenol (0.48 g) and triethylamine (0.28 g)
in 3 mL
dichloromethane is added dropwise. The reaction is placed in the dark and
stirred for 5 days. At
this time, the reaction is filtered through celite and concentrated under
reduced pressure.
Purification of the product by chromatography on silica gel affords 9c.
9d. 2',6'-diiodo-3-isopropyl-4-methoxy-4'-nitrilediphenyl ether: 2',6'-Diiodo-
3-isopropyl-
4-methoxy-4'-nitrilediphenyl ether (9c; 280 mg) is dissolved in 4 mL
dichloromethane and
cooled in a dry ice/acetone bath. To this solution is dropwise added 1.6 mL
boron tribromide (1
M in dichloromethane). The reaction is stirred overnight and allowed to reach
room
temperature. At this time, the reaction is poured onto 10 mL ice and water. To
this mixture is
added 10 mL ethyl acetate. The organic layer is separated and the aqueous
phase is extracted
twice with 10 mL ethyl acetate. The organic layers are combined, washed with
saturated sodium
chloride, dried over magnesium sulfate, and concentrated under reduced
pressure. Purification
of the product by chromatography on silica gel affords 9d.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
32
9e. 3,5-diiodo-4-(4'-hydroxy-3'-isopropylphenoxy)benzoic acid: To 2',6'-diiodo-
3-
isopropyl-4-methoxy-4'-nitrilediphenyl ether (9d; 95 mg) is added 1 mL 3 N
sodium hydroxide.
The sample is boiled and becomes homogeneous. At this time, the reaction is
cooled and
neutralized with 1 N HCI. A precipitate forms and is filtered off and washed
with hexane to
afford 9e.
Use of the Present Compounds
The methods of the present invention are performed by administering to a
mammal
(preferably a human) a compound having a structure as described herein and,
preferably, a
pharmaceutically-acceptable or cosmetically-acceptable carrier.
The compounds herein may be used for the treatment of such conditions as
treating hair
loss in mammals, including arresting and / or reversing hair loss and
promoting hair growth.
Such conditions may manifest themselves in, for example, alopecia, including
male pattern
baldness and female pattern baldness.
Preferably the compounds of the present invention are, as defined herein,
cardiac-
spanng.
Preferably, in the methods of the present invention, the compounds are
formulated into
pharmaceutical or cosmetic compositions for use in treatment or prophylaxis of
conditions such
as the foregoing. Standard pharmaceutical formulation techniques are used,
such as those
disclosed in Remin~ton's Pharmaceutical Sciences, Mack Publishing Company,
Easton, PA.
( 1990).
Typically, from about 5 mg to about 3000 mg, more preferably from about 5 mg
to
about 1000 mg, more preferably from about 10 mg to about 100 mg, of a compound
having
a structure as described herein is administered per day for systemic
administration. It is
understood that these dosage ranges are by way of example only, and that daily
administration can be adjusted depending on various factors. The specific
dosage of the
compound to be administered, as well as the duration of treatment, and whether
the
treatment is topical or systemic are interdependent. The dosage and treatment
regimen will
also depend upon such factors as the specific compound used, the treatment
indication, the
efficacy of the compound, the personal attributes of the subject (such as, for
example,
weight, age, sex, and medical condition of the subject), compliance with the
treatment
regimen, and the presence and severity of any side effects of the treatment.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
33
According to the present invention, the subject compounds are co-administered
with a
pharmaceutically-acceptable or cosmetically-acceptable carrier (herein
collectively described as
"carrier"). The term "carrier", as used herein, means one or more compatible
solid or liquid
filler diluents or encapsulating substances which are suitable for
administration to a mammal.
The term "compatible", as used herein, means that the components of the
composition are
capable of being commingled with a compound of the present invention, and with
each other, in
a manner such that there is no interaction which would substantially reduce
the efficacy of the
composition under ordinary use situations. Carriers must, of course, be of
sufficiently high
purity and sufficiently low toxicity to render them suitable for
administration to the animal,
preferably mammal (most preferably human), being treated. The carrier can
itself be inert or it
can possess pharmaceutical and / or cosmetic benefits of its own.
The compositions of this invention may be in any of a variety of forms,
suitable (for
example) for oral, rectal, topical, nasal, ocular or parenteral
administration. Of these,
topical and / or oral administration are especially preferred with topical
being most
preferred. Depending upon the particular route of administration desired, a
variety of
carriers well-known in the art may be used. These include solid or liquid
fillers, diluents,
hydrotropes, surface-active agents, and encapsulating substances. Optional
pharmaceutically-active or cosmetically-active materials may be included which
do not
substantially interfere with the activity of the compound of the present
invention. The
amount of carrier employed in conjunction with the compound is sufficient to
provide a
practical quantity of material for administration per unit dose of the
compound. Techniques
and compositions for making dosage forms useful in the methods of this
invention are
described in the following references: Modern Pharmaceutics, Chapters 9 and
10, Banker
& Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets
(1981);
and Ansel, Introduction to Pharmaceutical Dosage Forms, 2°d Ed.,
(1976).
Some examples of substances which can serve as Garners or components thereof
are
sugars, such as lactose, glucose and sucrose; starches, such as corn starch
and potato starch;
cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl
cellulose, and
methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants,
such as stearic acid
and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil,
cottonseed oil,
sesame oil, olive oil, corn oil and oil of theobroma; polyols such as
propylene glycol, glycerine,
sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers, such
as the TWEENS;
wetting agents, such sodium lauryl sulfate; coloring agents; flavoring agents;
tableting agents,
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
34
stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline;
and phosphate buffer
solutions.
The choice of a carrier to be used in conjunction with the subject compound is
typically
determined by the way the compound is to be administered.
In particular, carriers for systemic administration include sugars, starches,
cellulose
and its derivatives, malt, gelatin, talc. calcium sulfate, vegetable oils,
synthetic oils,
polyols, alginic acid, phosphate buffer solutions, emulsifiers, isotonic
saline, and pyrogen-
free water. Preferred carriers for parenteral administration include propylene
glycol, ethyl
oleate, pyrrolidone, ethanol, and sesame oil. Preferably, the carrier, in
compositions for
parenteral administration, comprises at least about 90% by weight of the total
composition.
Various oral dosage forms can be used, including such solid forms as tablets,
capsules, granules and bulk powders. These oral forms comprise a safe and
effective
amount, usually at least about 5%, and preferably from about 25% to about 50%,
of a
compound used in the present invention. Tablets can be compressed, tablet
triturates,
enteric-coated, sugar-coated, film-coated, or multiple-compressed, containing
suitable
binders, lubricants, diluents, disintegrating agents, coloring agents,
flavoring agents, flow-
inducing agents, and melting agents. Liquid oral dosage forms include aqueous
solutions,
emulsions, suspensions, solutions and/or suspensions reconstituted from non-
effervescent
granules, and effervescent preparations reconstituted from effervescent
granules, containing
suitable solvents, preservatives, emulsifying agents, suspending agents,
diluents,
sweeteners, melting agents, coloring agents and flavoring agents.
The carriers suitable for the preparation of unit dosage forms for oral
administration are
well-known in the art. Tablets typically comprise conventional
pharmaceutically-compatible
adjuvants as inert diluents, such as calcium carbonate, sodium carbonate,
mannitol, lactose and
cellulose; binders such as starch, gelatin and sucrose; disintegrants such as
starch, alginic acid
and croscarmelose; lubricants such as magnesium stearate, stearic acid and
talc. Glidants such
as silicon dioxide can be used to improve flow characteristics of the powder
mixture. Coloring .
agents, such as the FD&C dyes, can be added for appearance. Sweeteners and
flavoring agents,
such as aspartame, saccharin, menthol, peppermint, and fruit flavors, are
useful adjuvants for
chewable tablets. Capsules (including time release and sustained release
formulations) typically
comprise one or more solid diluents disclosed above. The selection of carrier
components
depends on secondary considerations like taste, cost, and shelf stability,
which are not critical for
the purposes of the subject invention, and can be readily made by a person
skilled in the art.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Orally administered compositions also include liquid solutions, emulsions,
suspensions,
powders, granules, elixirs, tinctures, syrups, and the like. The carriers
suitable for preparation of
such compositions are well known in the art. Typical components of carriers
for syrups, elixirs,
emulsions and suspensions include ethanol, glycerol, propylene glycol,
polyethylene glycol,
liquid sucrose, sorbitol and water. For a suspension, typical suspending
agents include methyl
cellulose, sodium carboxymethyl cellulose, AVICEL RC-591, tragacanth and
sodium alginate;
typical wetting agents include lecithin and polysorbate 80; and typical
preservatives include
methyl paraben and sodium benzoate. Peroral liquid compositions may also
contain one or more
components such as sweeteners, flavoring agents and colorants disclosed above.
Such compositions may also be coated by conventional methods, typically with
pH or
time-dependent coatings, such that the subject compound is released in the
gastrointestinal tract
in the vicinity of the desired topical application, or at various times to
extend the desired action.
Such dosage forms typically include, but are not limited to, one or more of
cellulose acetate
phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose
phthalate, ethyl cellulose,
Eudragit coatings, waxes and shellac.
Other compositions useful for attaining systemic delivery of the subject
compounds
include sublingual, buccal and nasal dosage forms. Such compositions typically
comprise one or
more of soluble filler substances such as sucrose, sorbitol and mannitol; and
binders such as
acacia, microcrystalline cellulose, carboxyrnethyl cellulose and hydroxypropyl
methyl cellulose.
Glidants, lubricants, sweeteners, colorants, antioxidants and flavoring agents
disclosed above
may also be included.
The compounds of the present invention may also be topically administered. The
carrier
of the topical composition preferably aids penetration of the present
compounds into the skin to
reach the environment of the hair follicle. Topical compositions of the
present invention may be
in any form including, for example, solutions, oils, creams, ointments, gels,
lotions, shampoos,
leave-on and rinse-out hair conditioners, milks, cleansers, moisturizers,
sprays, skin patches, and
the like.
Topical compositions containing the active compound can be admixed with a
variety of
carrier materials well known in the art, such as, for example, water,
alcohols, aloe vera gel,
allantoin, glycerine, vitamin A and E oils, mineral oil, propylene glycol, PPG-
2 myristyl
propionate, and the like.
Other materials suitable for use in topical carriers include, for example,
emollients,
solvents, humectants, thickeners and powders. Examples of each of these types
of materials,
which can be used singly or as mixtures of one or more materials, are as
follows:
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
36
Emollients, such as stearyl alcohol, glyceryl monoricinoleate, glyceryl
monostearate,
propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, iso-propyl
isostearate, stearic acid, iso-
butyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl
laurate, decyl oleate,
octadecan-2-ol, isocetyl alcohol, cetyl palmitate, dimethylpolysiloxane, di-n-
butyl sebacate, iso-
propyl myristate, iso-propyl palmitate, iso-propyl stearate, butyl stearate,
polyethylene glycol,
triethylene glycol, lanolin, sesame oil, coconut oil, arachis oil, castor oil,
acetylated lanolin
alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic
acid, isopropyl
linoleate, lauryl lactate, myristyl lactate, decyl oleate, and myristyl
myristate; propellants, such
as propane, butane, iso-butane, dimethyl ether, carbon dioxide, and nitrous
oxide; solvents, such
as ethyl alcohol, methylene chloride, iso-propanol, castor oil, ethylene
glycol monoethyl ether,
diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl
sulphoxide,
dimethyl formamide, tetrahydrofuran; humectants, such as glycerin, sorbitol,
sodium 2-
pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, and gelatin;
and powders, such as
chalk, talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide,
sodium polyacrylate,
tetra alkyl ammonium smectites, trialkyl aryl ammonium smectites, chemically
modified
magnesium aluminium silicate, organically modified montmorillonite clay,
hydrated aluminium
silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose,
and ethylene
glycol monostearate.
The compounds used in the present invention may also be administered in the
form of
liposome delivery systems, such as small unilamellar vesicles, large
unilamellar vesicles, and
multilamellar vesicles. Liposomes can be formed from a variety of
phospholipids, such as
cholesterol, stearylamine or phosphatidylcholines. A preferred formulation for
topical delivery
of the present compounds utilizes liposomes such as described in Dowton et
al., "Influence of
Liposomal Composition on Topical Delivery of Encapsulated Cyclosporin A: I. An
in vitro
Study Using Hairless Mouse Skin", S.T.P. Pharma Sciences, Vol. 3, pp. 404 -
407 (1993);
Wallach and Philippot, "New Type of Lipid Vesicle: Novasome ", Liposome
Technolo~y, Vol.
1, pp. 141 - 156 (1993); Wallach, U.S. Patent No. 4,911,928, assigned to Micro-
Pak, Inc., issued
March 27, 1990; and Weiner et al., U.S. Patent No. 5,834,014, assigned to The
University of
Michigan and Micro-Pak, Inc., issued November 10, 1998 (with respect to Weiner
et al., with a
compound as described herein administered in lieu of, or in addition to,
minoxidil).
The compounds of the present invention may also be administered by
iontophoresis.
See, e.~., Internet site www.unipr.it/area/dipfarm/erasmus/erasml4.html; Banta
et al.,
"Hydrogel-based Iontotherapeutic Delivery Devices for Transdermal Delivery of
Peptide/Protein
Drugs", Pharm. Res., Vol. 10 (5), pp. 697-702 (1993); F~, "Theoretical Model
of
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
37
Iontophoresis Utilized in Transdermal Drug Delivery", Pharmaceutical Acta
Helvetiae, Vol 70,
pp. 279-287 (1995); Gan ~rosa et al., "Modern Iontophoresis for Local Drug
Delivery", Int. J.
Pharm, Vol. 123, pp. 159-171 (1995); Green et al., "Iontophoretic Delivery of
a Series of
Tripeptides Across the Skin in vitro", Pharnt. Res., Vol 8, pp. 1121-1127
(1991); Jadoul et al.,
"Quantification and Localization of Fentanyl and TRH Delivered by
Iontophoresis in the Skin",
Int. J. Pharm., Vol. 120, pp. 221-8 (1995): O'Brien et al., "An Updated Review
of its Antiviral
Activity, Pharmacokinetic Properties and Therapeutic Efficacy", Drugs, Vol.
37, pp. 233-309
(1989); Parry et al., "Acyclovir Biovailability in Human Skin", J. Invest.
Dermatol., Vol. 98 (6),
pp. 856-63 (1992); Santi et al., "Drug Reservoir Composition and Transport of
Salmon
Calcitonin in Transdermal Iontophoresis", Pharm. Res., Vol 14 (1), pp. 63-66
(1997); Santi et
al., "Reverse Iontophoresis - Parameters Determining Electroosmotic Flow: I.
pH and Ionic
Strength", J. Control. Release, Vol. 38, pp. 159-165 (1996); Santi et al.,
"Reverse Iontophoresis
- Parameters Determining Electroosmotic Flow: II. Electrode Chamber
Formulation", J. Control.
Release, Vol. 42, pp. 29-36 (1996); Rao et al., "Reverse Iontophoresis:
Noninvasive Glucose
Monitoring in vivo in Humans", Pharm. Res., Vol. 12 (12), pp. 1869-1873
(1995); Thysman et
al., "Human Calcitonin Delivery in Rats by Iontophoresis", J. Pharm.
Pharmacol., Vol. 46, pp.
725-730 (1994); and Volpato et al., "Iontophoresis Enhances the Transport of
Acyclovir through
Nude Mouse Skin by Electrorepulsion and Electroosmosis", Pharm. Res., Vol. 12
(11), pp.
1623-1627 (1995).
The compositions used in the present invention may also optionally comprise an
activity
enhancer. The activity enhancer can be chosen from a wide variety of molecules
which can
function in different ways to enhance hair growth effects of a compound of the
present
invention. Particular classes of activity enhancers include other hair growth
stimulants and
penetration enhancers.
Non-limiting examples of other hair growth stimulants which may be used in the
compositions herein, including both systemic and topical compositions,
include, for example,
benzalkonium chloride, benzethonium chloride, phenol, estradiol,
diphenhydramine
hydrochloride, chlorpheniramine maleate, chlorophyllin derivatives,
cholesterol, salicylic acid,
cysteine, methionine, red pepper tincture, benzyl nicotinate, D,L - menthol,
peppermint oil,
calcium pantothenate, panthenol, castor oil, hinokitiol, prednisolone,
resorcinol,
monosaccharides and esterified monosaccharides, chemical activators of protein
kinase C
enzymes, glycosaminoglycan chain cellular uptake inhibitors, inhibitors of
glycosidase activity,
glycosaminoglycanase inhibitors, esters of pyroglutamic acid, hexosaccharic
acids or acylated
hexosaccharic acids, aryl-substituted ethylenes, N-acylated amino acids, and,
of course,
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
38
minoxidil or fmasteride. The most preferred activity enhancers are minoxidil
and finasteride,
most preferably minoxidil.
Non-limiting examples of penetration enhancers which may be used in the
compositions
herein include, for example, 2-methyl propan-2-ol, propan-2-ol, ethyl-2-
hydroxypropanoate,
hexan-2,5-diol, POE(2) ethyl ether, di(2-hydroxypropyl) ether, pentan-2,4-
diol, acetone, POE(2)
methyl ether, 2-hydroxypropionic acid, 2-hydroxyoctanoic acid, propan-1-ol,
1,4-dioxane,
tetrahydrofuran, butan-1,4-diol, propylene glycol dipelargonate,
polyoxypropylene 15 stearyl
ether, octyl alcohol, POE ester of oleyl alcohol, oleyl alcohol, lauryl
alcohol, dioctyl adipate,
dicapryl adipate, di-isopropyl adipate, di-isopropyl sebacate, dibutyl
sebacate, diethyl sebacate,
dimethyl sebacate, dioctyl sebacate, dibutyl suberate, dioctyl azelate,
dibenzyl sebacate, dibutyl
phthalate, dibutyl azelate, ethyl myristate, dimethyl azelate, butyl
myristate, dibutyl succinate,
didecyl phthalate, decyl oleate, ethyl caproate, ethyl salicylate, iso-propyl
palmitate, ethyl
laurate, 2-ethyl-hexyl pelargonate, iso-propyl isostearate, butyl laurate,
benzyl benzoate, butyl
benzoate, hexyl laurate, ethyl caprate, ethyl caprylate, butyl stearate,
benzyl salicylate, 2-
hydroxypropanoic acid, 2-hyroxyoctanoic acid, dimethyl sulphoxide, N,N-
dimethyl acetamide,
N,N-dimethyl formamide, 2-pyrrolidone, 1-methyl-2-pyrrolidone, 5-methyl-2-
pyrrolidone, 1,5-
dimethyl-2-pyrrolidone, 1-ethyl-2-pyrrolidone, phosphine oxides, sugar esters,
tetrahydrofurfural
alcohol, urea, diethyl-m-toluamide, and, 1-dodecylazacyloheptan-2-one.
In all of the foregoing, of course, the compounds used in the present methods
can be
administered alone or as mixtures, and the compositions may further include
additional drugs or
excipients as appropriate for the indication.
The present invention further relates to kits comprising a compound and / or
composition herein and information and / or instructions by words, pictures,
and / or the like,
that use of the kit will provide treatment for hair loss in mammals
(particularly humans)
including, for example, arresting and / or reversing hair loss and / or
promoting hair growth. In
addition or in the alternative, the kit may comprise a compound and / or
composition herein and
information and / or instructions regarding methods of application of the
compound and / or
composition, preferably with the benefit of treating hair loss in mammals.
Examples of Composition Administration
The following examples do not limit the invention, but provide guidance to the
skilled
artisan to perform the methods of the present invention. In each example, a
compound other
than the one mentioned may be substituted in the example by another having a
structure as
described herein with similar results.
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
39
Example A
A composition for topical administration is made, comprising:
Component Amount
Compound of Example5
3
Ethanol 57
Propylene Glycol 19
Dimethyl Isosorbide19
A human male subject suffering from male pattern baldness is treated by a
method of
this invention. Specifically, for 6 weeks, the above composition is daily
administered topically
to the subject.
Example B
A composition for topical administration is made according to the method of
Dowton et
al., "Influence of Liposomal Composition on Topical Delivery of Encapsulated
Cyclosporin A:
I. An in vitro Study Using Hairless Mouse Skin", S.T.P. Pharma Sciences, Vol.
3, pp. 404 - 407
(1993), using the compound of Example 2 in lieu of cyclosporin A and using the
Novasome 1 for
the non-ionic liposomal formulation.
A human male subject suffering from male pattern baldness is treated each day
with the
above composition. Specifically, for 6 weeks, the above composition is
administered topically
to the subject.
Example C
A shampoo is made, comprising:
Ex. Ex. Ex. Ex. C-4
C-1 C-2 C-3
Com onent
Ammonium Lau 1 Sulfate11.5 11.5 9.5 7.5
% % %
Ammonium Laureth 4 % 3 % 2 % 2
Sulfate
Cocamide MEA 2 % 2 % 2 % 2
Eth lene Gl col Distearate2 % 2 % 2 % 2
Cetyl Alcohol 2 % 2 % 2 % 2
Stea 1 Alcohol 1.2 1.2 1.2 1.2
% % %
Gl cerin 1 % 1 % 1 % 1
Pol uaternium 10 0.5 0.25 - -
% %
Pol uaternium 24 - - 0.5 0.25
%
Sodium Chloride 0.1 0.1 0.1 0.1
% % %
CA 02374262 2001-11-29
WO 00/72811 PCT/US00/05250
Sucrose Polyesters 3 % 3 % - -
of
Cottonate Fa Acid
Sucrose Polyesters 2 % 3 % - -
of
Behenate Fatty Acid
Polydimethyl Siloxane- - 3 % 2
Cocamino ro y1 Betaine- 1 % 3 % 3
Lau 1 Dimethyl Amine 1.5 1.5 1.5 1.5
Oxide % % %
Decyl Pol lucose - - 1 % 1
DMDM Hydantoin 0.15 0.15 0.15 0.15
% % %
Com ound of Exam 1e - 3 % 3 % -
1
Com ound of Exam 1e 6 % - - 6
4
Minoxidil 3 % 2
Phenoxyethanol 0.5 0.5 0.5 0.5
% % %
Fra ance 0.5 0.5 0.5 0.5
% % %
Water q.s. q.s. q.s. q.s.
A human subject sufferingfrom patternldness reated ethod
male ba is by a m of
t this
invention. Specifically, for 12 weeks, the above shampoo is used daily by the
subject.
