U.S. patent application number 13/783936 was filed with the patent office on 2013-07-11 for method for decreasing sebum production.
This patent application is currently assigned to Medicis Pharmaceutical Corporation. The applicant listed for this patent is Catherine R. Kostlan, Raj Neil Raheja, Meera Tugnait, Kimberly Wade. Invention is credited to Catherine R. Kostlan, Raj Neil Raheja, Meera Tugnait, Kimberly Wade.
Application Number | 20130178446 13/783936 |
Document ID | / |
Family ID | 34435045 |
Filed Date | 2013-07-11 |
United States Patent
Application |
20130178446 |
Kind Code |
A1 |
Kostlan; Catherine R. ; et
al. |
July 11, 2013 |
METHOD FOR DECREASING SEBUM PRODUCTION
Abstract
The present invention is directed to the topical application of
the malonamide ACAT inhibitors described by Formula I. Other
aspects of the invention are directed to topical formulations of
these diamides, their use to treat sebaceous gland disorders and
their use to alleviate oily skin.
Inventors: |
Kostlan; Catherine R.;
(Saline, MI) ; Raheja; Raj Neil; (Ann Arbor,
MI) ; Tugnait; Meera; (Ann Arbor, MI) ; Wade;
Kimberly; (Brighton, MI) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Kostlan; Catherine R.
Raheja; Raj Neil
Tugnait; Meera
Wade; Kimberly |
Saline
Ann Arbor
Ann Arbor
Brighton |
MI
MI
MI
MI |
US
US
US
US |
|
|
Assignee: |
Medicis Pharmaceutical
Corporation
Scottsdale
AZ
|
Family ID: |
34435045 |
Appl. No.: |
13/783936 |
Filed: |
March 4, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12568763 |
Sep 29, 2009 |
8409595 |
|
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13783936 |
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10958306 |
Oct 5, 2004 |
7615230 |
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12568763 |
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60509984 |
Oct 9, 2003 |
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Current U.S.
Class: |
514/119 ;
514/239.5; 514/275; 514/352; 514/381; 514/459; 514/460; 514/471;
514/539; 514/596; 514/616; 514/621 |
Current CPC
Class: |
A61K 31/167 20130101;
A61K 31/341 20130101; A61P 17/00 20180101; A61K 31/16 20130101;
A61Q 19/00 20130101; A61K 8/49 20130101; A61P 17/08 20180101; A61K
8/4946 20130101; A61K 8/4926 20130101; A61K 31/664 20130101; A61K
8/494 20130101; A61K 31/17 20130101; A61K 31/4402 20130101; A61P
43/00 20180101; A61K 2800/782 20130101; A61K 31/5375 20130101; A61K
8/4973 20130101; A61K 8/42 20130101; A61K 31/41 20130101; A61K
8/498 20130101; A61K 31/216 20130101; A61K 31/44 20130101; A61Q
19/008 20130101; A61K 31/505 20130101; A61K 31/351 20130101; A61P
17/10 20180101 |
Class at
Publication: |
514/119 ;
514/239.5; 514/275; 514/352; 514/381; 514/459; 514/460; 514/471;
514/539; 514/596; 514/616; 514/621 |
International
Class: |
A61K 31/664 20060101
A61K031/664; A61K 31/505 20060101 A61K031/505; A61K 31/4402
20060101 A61K031/4402; A61K 31/167 20060101 A61K031/167; A61K
31/351 20060101 A61K031/351; A61K 31/341 20060101 A61K031/341; A61K
31/216 20060101 A61K031/216; A61K 31/17 20060101 A61K031/17; A61K
31/5375 20060101 A61K031/5375; A61K 31/41 20060101 A61K031/41 |
Claims
1-23. (canceled)
24. A method for treating a dermal disease comprising topically
administering to a patient in need thereof an effective amount of a
compound of the formula I: ##STR00036## in which R.sup.1 and
R.sup.2 are each independently C.sub.1-6 alkyl; X is CH.sub.2;
R.sup.3 is C.sub.1-6 alkyl or pyridyl; p is 1; R.sup.4 is selected
from the group consisting of C.sub.1-6 alkyl, C.sub.1-6 alkoxy, and
SR.sup.7; R.sup.7 is C.sub.1-6 alkyl; or a pharmaceutically
acceptable salt thereof.
25. A method for treating a dermal disease comprising topically
administering to a patient in need thereof an effective amount of a
compound of the formula IA: ##STR00037## in which R.sup.3 is
C.sub.1-6 alkyl or pyridyl; R.sup.4 is selected from the group
consisting of C.sub.1-6 alkyl, C.sub.1-6 alkoxy, and SR.sup.7;
R.sup.7 is C.sub.1-6 alkyl; or a pharmaceutically acceptable salt
thereof.
26. A method of claim 24, wherein said compound is selected from
the group consisting of:
N'-[2,6-bis(1-methylethyl)phenyl]-N-(1-methylethyl)-N-[[4-(methylthio)phe-
nyl]methyl]-propanediamide or pharmaceutically acceptable salt
thereof, and
N-[2,6-bis(1-methylethyl)phenyl]-].beta.-[[(4-methoxyphenyl)methyl](2-
-pyridinyl)amino]-].beta.]-oxo-propanamide or pharmaceutically
acceptable salt thereof.
27. A method for treating a dermal disease comprising the topical
administration of an effective amount of
N-benzyl-N'-(2,6-diisopropyl-phenyl)-N-isopropyl-malonamide, or a
pharmaceutically acceptable salt thereof, to a patient in need
thereof.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of U.S. Provisional
Application 60/509,984 filed Oct. 9, 2003.
FIELD OF THE INVENTION
[0002] The present invention is directed to the topical application
of a class of diamide ACAT inhibitors. Other aspects of the
invention are directed to topical formulations of these diamides,
their use to treat sebaceous gland disorders and their use to
alleviate oily skin.
BACKGROUND OF THE INVENTION
[0003] Human skin is composed of three primary layers, the stratum
corneum, the epidermis, and the dermis. The outer layer is the
stratum corneum. Its primary function is to serve as a barrier to
the external environment. Lipids are secreted to the surface of the
stratum corneum. These lipids decrease the stratum corneum's water
permeability. Sebum typically constitutes 95% of these lipids.
Abramovits et al, Dermatologic Clinics, Vol 18, Number 4, Oct.
2000.
[0004] Sebum is produced in the sebaceous glands. These glands are
present over most of the surface of the body. The highest
concentration of these glands occurs on the scalp, the forehead and
the face. Despite the important physiological role that sebum
plays, many individuals experience excess sebum production,
especially in the facial area. Excess sebum is associated with an
increased incidence of acne. Even in individuals without acne,
sebum can make the skin look greasy, decreasing its attractiveness.
Abramovits et al, supra.
[0005] Current treatments for excess sebum are less than optimal.
Accutane (isotretinoin) reduces sebum secretion by up to 90%.
However, isotretinoin is associated with a number of serious side
effects. It causes serious birth defects and is contraindicated in
women of childbearing age. Thus, isotretinoin is only utilized for
severe acne. It is inappropriate to use this drug merely as a
cosmetic aid.
[0006] Acyl CoA cholesterol acyl transferase (ACAT) inhibitors were
initially evaluated to treat elevated cholesterol. U.S. Pat. No.
6,133,326 discloses that ACAT inhibitors also reduce the secretion
of sebum. While the '326 patent is a valuable contribution to the
art, such treatments are not commercially available at the present
time. Currently, the most practical means of alleviating excess
sebum is frequent washings. Thus, a need exists in the art for new
treatments that will reduce the secretion of sebum by the sebaceous
glands.
SUMMARY OF THE INVENTION
[0007] A new method for decreasing the secretion of sebum, by the
sebaceous glands, has been discovered. A class of ACAT inhibitors
that exhibit superior activity in the inhibition of sebum secretion
has been discovered. These ACAT inhibitors may be represented by
Formula I:
##STR00001##
in which R.sup.1 and R.sup.2 are each independently represented by
hydrogen, C.sub.1-8 alkyl, C.sub.1-8 alkoxy, halogen, hydroxy,
trifluoromethyl, trifluoromethoxy, cyano, NR.sup.5R.sup.6, or
SR.sup.7; X is represented by --CR.sup.8R.sup.9--(CH.sub.2).sub.n;
R.sup.3 is represented by hydrogen, C.sub.1-8 alkyl,
--(CH.sub.2).sub.q-Ph, or --(CH.sub.2).sub.q-M; p is represented by
an integer from 1 to 4; R.sup.4 is represented by a substituent
selected from the group consisting of hydrogen, C.sub.1-6 alkyl,
C.sub.1-6 alkoxy, halogen, hydroxy, trifluoromethyl,
trifluoromethoxy, cyano, NR.sup.5R.sup.6, and SR.sup.7; R.sup.5,
R.sup.6, R.sup.7, R.sup.8, and R.sup.9 are each independently
represented by hydrogen or C.sub.1-6 alkyl; Ph is represented by a
phenyl ring which may be optionally substituted; M is represented
by a 5- or 6-membered heteroaryl ring, containing 1 hetero-atom
selected from the group N, S, or O; n and q are each independently
represented by an integer from 0-4; a pharmaceutically acceptable
salt thereof, or a prodrug thereof.
[0008] The compounds of Formula I may be administered to a patient
to decrease the amount of sebum secreted by their sebaceous glands.
Typically, the compounds will be administered topically to the
areas exhibiting excess sebum production. Decreasing sebum
secretion will alleviate a number of dermatological disorders and
cosmetic complaints. These conditions include oily skin, oily hair,
shiny skin, acne, and seborrheic dermatitis.
[0009] The invention is also directed to pharmaceutical
compositions containing at least one of the compounds of Formula I
in admixture with a carrier suitable for topical administration. In
a further embodiment, the invention is directed to an article of
manufacture containing a compound of Formula I, packaged for retail
distribution, in association with instructions advising the
consumer on how to use the compound to alleviate a condition
associated with excess sebum production. An additional embodiment
is directed to the use of a compound of Formula I as a diagnostic
agent to detect inappropriate sebum production. Other aspects of
the invention are directed to the use of a compound of Formula I in
the manufacture of a medicament for seborrhea.
DETAILED DESCRIPTION OF THE INVENTION
[0010] The headings within this document are only being utilized
expedite its review by the reader. They should not be construed as
limiting the invention or claims in any manner.
[0011] A) Definitions and Exemplification
[0012] As used throughout this application, including the claims,
the following terms have the meanings defined below, unless
specifically indicated otherwise. The plural and singular should be
treated as interchangeable, other than the indication of number:
[0013] a. "C.sub.1-C.sub.6 alkyl" refers to a branched or straight
chained alkyl group containing from 1 to 6 carbon atoms, such as
methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, n-pentyl,
isopentyl, n-hexyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, etc.
[0014] b. "C.sub.1-C.sub.6 alkoxy" refers to a straight or branched
chain alkoxy group containing from 1 to 6 carbon atoms, such as
methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy,
n-pentoxy, n-hexyloxy, etc. [0015] c. "halogen" refers to a
chlorine, fluorine or bromine atom. [0016] d. "optionally
substituted phenyl" refers to a phenyl (--C.sub.6H.sub.5) which may
be substituted with up to 3 substituents, each substituent is
independently selected from the group consisting of C.sub.1 alkyl,
C.sub.1-6 alkoxy, halogen, hydroxy, trifluoromethyl,
trifluoromethoxy, cyano, NR.sup.5R.sup.6, or SR.sup.7. These
substituents may be the same or different and may be located at any
of the ortho, meta, or para positions. [0017] e. "heteroaryl"
refers to aromatic ring having a single heteroatom selected from
oxygen, nitrogen and sulfur. More specifically, it refers to a 5-,
or 6-, membered ring containing 1 nitrogen atom, 1 oxygen atom, or
1 sulfur atom. The 5-membered ring has 2 double bonds and the
6-membered ring has 3 double bonds. Examples of such heteroaryl
ring systems include, but is not limited to pyrrolyl, furanyl,
thiophenyl, and pyridinyl. [0018] f. "pharmaceutically acceptable
salts" is intended to refer to either pharmaceutically acceptable
acid addition salts or "pharmaceutically acceptable basic addition
salts" depending upon actual structure of the compound. [0019] g.
"pharmaceutically acceptable acid addition salts" is intended to
apply to any non-toxic organic or inorganic acid addition salt of
the base compounds represented by Formula I or any of its
intermediates. Illustrative inorganic acids which form suitable
salts include hydrochloric, hydrobromic, sulphuric, and phosphoric
acid and acid metal salts such as sodium monohydrogen
orthophosphate, and potassium hydrogen sulfate. Illustrative
organic acids, which form suitable salts include the mono-, di-,
and tricarboxylic acids. Illustrative of such acids are for
example, acetic, glycolic, lactic, pyruvic, malonic, succinic,
glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic,
hydroxymaleic, benzoic, hydroxy-benzoic, phenylacetic, cinnamic,
salicylic, 2-phenoxybenzoic, p-toluenesulfonic acid, and sulfonic
acids such as methane sulfonic acid and 2-hydroxyethane sulfonic
acid. Such salts can exist in either a hydrated or substantially
anhydrous form. In general, the acid addition salts of these
compounds are soluble in water and various hydrophilic organic
solvents, and which in comparison to their free base forms,
generally demonstrate higher melting points. [0020] h.
"pharmaceutically acceptable basic addition salts" is intended to
apply to any non-toxic organic or inorganic basic addition salts of
the compounds represented by Formula I, or any of its
intermediates. Illustrative bases which form suitable salts include
alkali metal or alkaline-earth metal hydroxides such as sodium,
potassium, calcium, magnesium, or barium hydroxides; ammonia, and
aliphatic, alicyclic, or aromatic organic amines such as
methylamine, dimethylamine, trimethylamine, and picoline. [0021] i.
"prodrug" refers to compounds that are rapidly transformed in vivo
to yield the parent compound of the above formulas, for example, by
hydrolysis in blood. A thorough discussion is provided in T.
Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems," Vol.
14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in
Drug Design, ed. Edward B. Roche, American Pharmaceutical
Association and Pergamon Press, 1987, both of which are
incorporated herein by reference. [0022] j. "compound of Formula
I", "compounds of the invention" and "compounds" are used
interchangeably throughout the application and should be treated as
synonoms. [0023] k. "patient" refers to warm blooded animals such
as, for example, guinea pigs, mice, rats, gerbils, cats, rabbits,
dogs, monkeys, chimpanzees, and humans. [0024] l. "treat" refers to
the ability of the compounds to either relieve, alleviate, or slow
the progression of the patient's disease (or condition) or any
tissue damage associated with the disease.
[0025] Some of the compounds of Formula I will exist as optical
isomers. Any reference in this application to one of the compounds
represented by Formula I is meant to encompass either a specific
optical isomer or a mixture of optical isomers (unless it is
expressly excluded). The specific optical isomers can be separated
and recovered by techniques known in the art such as chromatography
on chiral stationary phases or resolution via chiral salt formation
and subsequent separation by selective crystallization.
Alternatively utilization of a specific optical isomer as the
starting material will produce the corresponding isomer as the
final product.
[0026] In addition, the compounds of the present invention can
exist in unsolvated as well as solvated forms with pharmaceutically
acceptable solvents such as water, ethanol, and the like. In
general, the solvated forms are considered equivalent to the
unsolvated forms for the purposes of the present invention.
[0027] Any reference in this application to a compound of Formula
I, is intended to cover the compounds individually, as mixtures, as
salts, as solvates or any combination thereof.
[0028] All of the compounds of Formula I have at least two phenyl
rings, as depicted immediately below:
##STR00002##
[0029] Ring A may be unsubstituted, or it may be substituted with
one or two substituents as defined by R.sup.1 and R.sup.2. R.sup.1
and R.sup.2 may be represented by identical substituents, or
different substituents. In one embodiment, R.sup.1 and R.sup.2 are
each represented by isopropyl moieties and are both located at the
ortho positions of the phenyl ring.
[0030] Ring B may also be optionally substituted, as listed for
R.sup.4. R.sup.4 may represent up to 3 substituents, other than
hydrogen, as described above. These substituents may be located at
any of the ortho, meta, or para positions.
[0031] R.sup.3 may also be represented may a phenyl ring or a
phenylalkylene moiety. Any such phenyl ring may also be substituted
with up to 3 substituents, as described above. They may be located
at any of the ortho, meta, or para positions.
[0032] R.sup.3 may also be represented by a heteroaryl ring or by a
heteroarylalkylene moiety. The heteroaryl ring may be attached to
the indicated nitrogen atom by any carbon atom of the heteroaryl
ring. Likewise, if q is 1, 2, 3, or 4, then the heteroaryl ring may
be bonded to the alkylene moiety via any of its carbon atoms.
[0033] In a further embodiment of the invention, Formula IA
exemplifies a subgenus of Formula I, particularly useful for
topical application.
##STR00003##
[0034] In Formula IA, R.sup.1 and R.sup.2 are each isopropyl
(ortho), p is 1 and X is methylene, as exemplified above; R.sup.3
is represented by C.sub.1 alkyl or heteroaryl, (more typically
isopropyl, or pyridyl) and R.sup.4 is as defined in Formula I.
[0035] More specific examples of compounds of Formula la include:
[0036] a)
N-benzyl-N'-(2,6-diisopropyl-phenyl)-N-isopropyl-malonamide; [0037]
b)
N'-[2,6-bis(1-methylethyl)phenyl]-N-(1-methylethyl)-N-[[4-(methylthio)phe-
nyl]methyl]-propanediamide and; [0038] c)
N-[2,6-bis(1-methylethyl)phenyl]-].beta.-[[(4-methoxyphenyl)methyl](2-pyr-
idinyl)amino]-].beta.-oxo-propanamide.
[0039] B) Synthesis
[0040] The compounds of Formula I have previously been described in
the literature. The reader's attention is directed to European
Patent Application Number 0 433 662, which is hereby incorporated
by reference. The '662 application discloses that the compounds of
Formula I have acyl-coenzyme A cholesterol acyltransferase (ACAT)
inhibitory activity. The '662 application discloses that these
compounds can be used to lower elevated cholesterol levels and to
treat atherosclerosis. The '662 application does not disclose using
these compounds to decrease sebum secretion.
[0041] The '662 application discloses how to prepare the compounds
of Formula I. The reader's attention is directed to pages 7-20
where methods for synthesizing these compounds are described.
Methods for preparing pharmaceutically acceptable salts of these
compounds are described on page 6 of the specification.
[0042] C) Medical and Cosmetic Uses
[0043] Inhibition of acyl-CoA cholesterol acyl transferase (ACAT)
blocks the esterification of free cholesterol-to-cholesterol
esters. Cholesterol esters are the primary transportation and
storage form of cholesterol in animals. In the intestines, ACAT
inhibitors have been shown to inhibit the absorption of cholesterol
from the gut. In the liver, inhibition of ACAT has been shown to
decrease the formation and secretion of cholesterol-containing
lipoproteins by decreasing the cholesterol ester mass of the
lipoprotein core. For these reasons, ACAT inhibitors have
previously been evaluated as a means to lower serum cholesterol
levels.
[0044] Dermal sebaceous glands are holocrine glands that secrete a
mixture of lipids known as sebum. Sebum is composed of
triglycerides, wax, sterol esters and squalene. There is
considerable variation in the composition of human sebum based on
individual variables such as age, sex, diet, and disease. Sebum is
produced in the acinar cells of sebaceous glands, accumulates as
those cells age and migrates towards the center of the gland. At
maturation, the acinar cells lyse and release sebum into the
lumenal duct, from which the sebum is secreted.
[0045] Formation of sebum is regulated by a variety of hormones
that act primarily to regulate the rate of lipid metabolism. Waxes
and sterols are converted, within acinar cells, to a stable ester
form for storage via the activity of a variety of acyl and fatty
acid transferases. These esters are then stored in lipid droplets
within the acinar cells prior to release.
[0046] The compounds of formula I block the conversion of free
cholesterol-to-cholesterol ester, leading to increased levels of
free cholesterol within the acinar cells. While the cellular
mechanism is not fully understood at the present time, the acinar
cells produce less sebum when contacted with an ACAT inhibitor.
[0047] Thus, the compounds of formula I inhibit the secretion of
sebum and thus reduce the amount of sebum on the surface of the
skin. The compounds can be used to treat a variety of dermal
diseases such as acne or seborrheic dermatitis.
[0048] In addition to treating diseases associated with excess
sebum production, the compounds can also be used to achieve a
cosmetic effect. Some consumers believe that they are afflicted
with overactive sebaceous glands. They feel that their skin is oily
and thus unattractive. These individuals can utilize the compounds
of Formula I to decrease the amount of sebum on their skin.
Decreasing the secretion of sebum will alleviate oily skin in
individuals afflicted with such conditions.
[0049] In order to exhibit the biological effects described above,
the compounds need to be administered in a quantity sufficient to
inhibit production and/or secretion of sebum by the sebaceous
glands and acinar cells. This amount can vary depending upon the
particular disease/condition being treated, the severity of the
patient's disease/condition, the patient, the particular compound
being administered, the route of administration, and the presence
of other underlying disease states within the patient, etc. When
administered systemically, the compounds typically exhibit their
effect at a dosage range of from about 0.1 mg/kg/day to about 100
mg/kg/day for any of the diseases or conditions listed above.
Repetitive daily administration may be desirable and will vary
according to the conditions outlined above.
[0050] The compounds of the present invention may be administered
by a variety of routes. They are effective if administered orally.
The compounds may also be administered parenterally (i.e.
subcutaneously, intravenously, intramuscularly, intraperitoneally,
or intrathecally), rectally, or topically.
[0051] In a typical embodiment, the compounds are administered
topically. Topical administration is especially appropriate for
acne and for cosmetic indications. The compound will be applied to
those areas of the skin afflicted with excess sebum production. The
dose will vary, but as a general guideline, the compound will be
present in a dermatologically acceptable carrier in an amount of
from 0.01 to 10 w/w % and the dermatological preparation will be
applied to the affected area from 1 to 4 times daily.
"Dermatologically acceptable" refers to a carrier which may be
applied to the skin, hair or scalp, and which will allow the drug
to diffuse to the site of action. (i.e. the sebaceous glands and/or
the acinar cells).
[0052] D) Co-Administration
[0053] In a further embodiment of the invention, the compounds of
Formula I can be co-administered with other compounds to further
enhance their activity, or to minimize potential side effects. For
example, antibiotics, such as tetracycline and clindamycin, have
been used to alleviate acne. The antibiotic eradicates the
microorganism, Propionbacterium acnes, leading to a reduction in
the patient's acne. The compounds of Formula I can be
co-administered with any antibiotic suitable for the treatment of
acne.
[0054] Retinoids, such as isotretinoin, have been shown to decrease
sebum production and are used to treat acne. These retinoids can be
co-administered with a compound of Formula I in order to decrease
sebum production and/or to treat acne.
[0055] Estrogen and progesterone have each been shown to decrease
sebum production. These compounds, or any synthetic agonist of the
estrogen or progesterone receptor, may be co-administered with a
compound of formula I in order to decrease sebum production.
[0056] Anti-androgens have been shown to decrease sebum secretion.
Anti-androgens can work by a number of different mechanisms. For
example, some compounds block the conversion of testosterone to
5-.alpha.-dihydrotestosterone, which is responsible for the
biological effect in many tissues. 5-Alpha-reductase inhibitors,
such as finasteride, have been shown to decrease sebum production.
Finasteride is commercially available from Merck under the trade
name Propecia.RTM.. Examples of other 5-.alpha.-reductase
inhibitors include dutasteride (Glaxo Smithkline). Other
anti-androgens are antagonists of the androgen receptor. For
example, androgen antagonists, such as flutamide, have been
reported to decrease sebum production. Such compounds can be
co-administered with the compounds of Formula I to decrease sebum
production.
[0057] As used in this application, co-administered refers to
administering a compound of Formula I with a second medicinal,
typically having a differing mechanism of action, using a dosing
regimen that promotes the desired result. This can refer to
simultaneous dosing, dosing at different times during a single day,
or even dosing on different days. The compounds can be administered
separately or can be combined into a single formulation. Techniques
for preparing such formulations are described below.
[0058] E. Cosmetic and Pharmaceutical Formulations
[0059] If desired, the compounds can be administered directly
without any carrier. However, to ease administration, they will
typically be formulated into pharmaceutical carriers
[0060] For oral administration, the compounds can be formulated
into solid or liquid preparations such as capsules, pills, tablets,
lozenges, melts, powders, suspensions, or emulsions. Solid unit
dosage forms can be capsules of the ordinary gelatin type
containing, for example, surfactants, lubricants and inert fillers
such as lactose, sucrose, and cornstarch or they can be sustained
release preparations.
[0061] In another embodiment, the compounds can be tableted with
conventional tablet bases such as lactose, sucrose, and cornstarch
in combination with binders, such as acacia, cornstarch, or
gelatin, disintegrating agents such as potato starch or alginic
acid, and a lubricant such as stearic acid or magnesium stearate.
Liquid preparations are prepared by dissolving the active
ingredient in an aqueous or non-aqueous pharmaceutically acceptable
solvent, which may also contain suspending agents, sweetening
agents, flavoring agents, and preservative agents as are known in
the art.
[0062] For parenteral administration the compounds may be dissolved
in a physiologically acceptable pharmaceutical carrier and
administered as either a solution or a suspension. Illustrative of
suitable pharmaceutical carriers are water, saline, dextrose
solutions, fructose solutions, ethanol, or oils of animal,
vegetative, or synthetic origin. The pharmaceutical carrier may
also contain preservatives, buffers, etc., as are known in the art.
When the compounds are being administered intrathecally, they may
also be dissolved in cerebrospinal fluid as is known in the
art.
[0063] Typically however, the compounds will be incorporated into
formulations suitable for topical administration. Any of the
topical formulations known in the art may be used. Examples of such
topical formulations include lotions, sprays, creams, ointments,
salves, gels, etc. Actual methods for preparing topical
formulations are known or apparent to those skilled in the art, and
are described in detail in Remington's Pharmaceutical Sciences,
1990 (supra); and Pharmaceutical Dosage Forms and Drug Delivery
Systems, 6th ed., Williams & Wilkins (1995).
[0064] In a further embodiment, the formulations described above
may be packaged for retail distribution (i.e., a kit or article of
manufacture). The package will contain instructions advising the
patient how to use the product to alleviate conditions such as
acne, oily skin, etc. Such instructions may be printed on the box,
may be a separate leaflet or printed on the side of the container
holding the formulation, etc.
[0065] The compounds of Formula I may also be admixed with any
inert carrier and utilized in laboratory assays in order to
determine the concentration of the compounds within the serum,
urine, etc., of the patient as is known in the art. The compounds
may also be used as a research tool.
[0066] While the invention has been described in connection with
specific embodiments thereof, it will be understood that it is
capable of further modifications and this application is intended
to cover any variations, uses, or adaptations of the invention
following, in general, the principles of the invention and
including such departures from the present disclosure as come
within known or customary practice within the art to which the
invention. The following examples and biological data are being
presented in order to further illustrate the invention. This
disclosure should not be construed as limiting the invention in any
manner.
EXAMPLE I
[0067] Luderschmidt et al describes an animal model for testing
whether compounds are capable of modulating sebum secretion. Arch.
Derm. Res. 258, 185-191 (1977). This model uses Syrian hamsters,
whose ears contain sebaceous glands. Compounds can be administered
to these animals to determine if a test agent is capable of
modulating sebum production
[0068] A series of compounds known to inhibit ACAT were screened
using methods analogous to those of Luderschmidt et al. Table IA
shows the results obtained with selected diamides encompassed by
Formula I above. Table IB shows the results obtained with a series
of diamides not encompassed by Formula I. Formula IC shows the
results obtained with other potent ACAT inhibitors that are not
diamides.
[0069] Tables IA-IC also reports the affinity of the compound for
rat ACAT, measured as an IC.sub.50. These values were determined by
measuring the ACAT-mediated transfer of tritiated oleic acid from
acyl-CoA to cholesterol to give labeled cholesteryl oleate. The
source of ACAT activity was homogenates of rat intestinal tissue.
Predetermined concentrations of: intestinal homogenate containing
endogenous cholesterol, 2) test compound, and 3) [1-.sup.14C]
oleolyl-CoA were contacted for a predetermined time. The reaction
was quenched and the results were determined by thin layer
chromatography. Analogous assays using rabbit intestine were
described by Roth et al in J. Med Chem. 35:1609-1617(1992).
[0070] Testing for sebum inhibition was carried out in the
following manner. Male Syrian hamsters aged 9 to 10 weeks were
introduced into the laboratory environment and acclimated for 2
weeks prior to use in the study. Each group consisted of 5 animals
and were run in parallel with vehicle and positive controls. Prior
to administration, 10 mg of each compound was dissolved in 1 mL of
Universal solvent (ethanol/propylene glycol (70/30% v/v) to achieve
a final concentration of 1 w/v %.
[0071] Animals were dosed topically twice daily, five days a week,
for 4 weeks. Each dose consisted of 25 micro liters of vehicle
control or drug. The dose was applied to the ventral surfaces of
both the right and left ears. All animals were sacrificed
approximately 18-24 hours after the final dose. The right ears were
collected from each animal and used for sebum analysis.
[0072] The ears were prepped for HPLC analysis in the following
manner. One 8 mm distal biopsy punch was taken, just above the
anatomical "V" mark in the ear to normalize the sample area. The
punch was pulled apart. The ventral biopsy surface (the area where
the topical dose was directly applied to the sebaceous glands) was
retained for testing and the dorsal surface of the biopsy punch was
discarded.
[0073] Tissue samples were blown with N.sub.2 gas and stored at
-80.degree. C. under nitrogen until HPLC analysis. In addition to
ear samples, an aliquot of each drug and vehicle (at least 250 ul)
was also stored at -80.degree. C. for inclusion in the HPLC
analysis.
[0074] HPLC analysis was carried out on an extract of the tissue
sample. Tissue samples were contacted with 3 ml of solvent (a 4:1
admixture of 2,2,4-trimethylpentane and isopropyl alcohol). The
mixture was shaken for 15 minutes and stored overnight at room
temperature, protected from light. The next morning 1 milliliter of
water was added to the sample and shaken for 15 minutes. The sample
was then centrifuged at approximately 1500 rpm for 15 minutes. Two
ml of the organic phase (top layer) was transferred to a glass
vial, dried at 37.degree. C., under nitrogen, for approximately 1
hour, and then lyophilized for approximately 48 hours. The samples
were then removed from the lyophilizer and each vial was
reconstituted with 600 .mu.l of solvent A
(trimethylpentane/tetrahydrofuran (99:1). The samples were then
recapped and vortexed for 5 minutes. 200 .mu.l of each sample was
then transferred to a pre-labeled 200 .mu.l HPLC vial with 200
.mu.L glass inserts. The HPLC vials were placed in the autosampler
tray for the Agilent 1100 series HPLC unit. The Agilent 1100 HPLC
system consisted of a thermostated autosampler, a quarternary pump,
a column heater, and an A/D interface module. All components were
controlled by Agilent ChemStation software. A Waters Spherisorb S3W
4.6.times.100 mm analytical column was maintained at 30.degree. C.
by the Agilent column heater unit. The HPLC autosampler was
programmed to maintain the sample temperature at 20.degree. C.
throughout the run.
[0075] 10 uL of each sample was injected in triplicate into the
column. Two solvents were used for the solvent gradient. Solvent A
was an admixture of trimethylpentane and tetrahydrofuran (99:1).
Solvent B was ethylacetate. The gradient utilized is described in
the table below:
TABLE-US-00001 Time (min) Solv A (%) Solv B (%) Flow (mL/min) 0 99
1 2 2 96 4 2 6 60 40 2 7 5 95 2 10 5 95 2 10.1 99 1 2
[0076] The Sedex 75 Evaporative Light Scattering Detector (ELSD)
was operated at 45.degree. C. with a gain of 5, and N.sub.2
pressure maintained at 3.1 bar. Analog signal obtained by the
instrument was sent to the Agilent A/D interface module where it
was converted to a digital output. The conversion was based on a
10000 mAU/volt set point and the data rate was set at 10 Hz (0.03
min). The resulting digital output was then feed into the Agilent
ChemStation software for integration of the peak area. The results
of the HPLC analysis are reported below in Tables IA-C.
[0077] The results are reported as the reduction in cholesterol
ester (CE) and wax ester (WE) production, when compared to the
vehicle control. A negative number indicates that the ACAT
inhibitor actually increased production of sebum.
TABLE-US-00002 TABLE IA Compounds of Invention ACAT Inhibition
Change vs. relevant Compound Information (IC 50) vehicle control %
% Compound Reduction Reduction Sum Number Molecular Structure IAI
(nM) in CE in WE CE + WE 1 ##STR00004## 15 94% 80% 174% 1 * (tested
multiple times) ##STR00005## 15 94% 84% 178% 2 ##STR00006## 26 65%
31% 96% 3 ##STR00007## 31 51% 33% 84%
TABLE-US-00003 TABLE IB Comparative Examples- ACAT Inhibition
Change vs relevant Compound Information (IC 50) vehicle control % %
Compound Reduction Reduction Sum Number Molecular Structure IAI
(nM) in CE in WE CE + WE 4 ##STR00008## 170 -1% -18% -19% 5
##STR00009## 61 32% -4% 28% 6 ##STR00010## 72 23% -1% 22% 7
##STR00011## 26 -67% -84% -151% 8 ##STR00012## 14 -27% -70%
-97%
TABLE-US-00004 TABLE IC ACAT Inhibition Change vs relevant Compound
Information (IC 50) vehicle control % % Compound Reduction
Reduction Sum Number Molecular Structure IAI (nM) in CE in WE CE +
WE 9 ##STR00013## 44 7% 2% 9% 10 ##STR00014## 8 -105% -147% -252%
11 ##STR00015## 8.5 -5% -6% -11% 12 ##STR00016## 15 48% 44% 92% 13
##STR00017## 42 -7% -7% -14% 14 ##STR00018## 6 -8% -15% -23% 15
##STR00019## 3.4 24% 14% 38% 16 ##STR00020## 26 -1% 0% -1% 17
##STR00021## 16 -56% -64% -120% 18 . ##STR00022## 17 -23% -49% -72%
19 ##STR00023## 12 -6% 1% -5% 20 ##STR00024## 32 4% -1% 3% 21
##STR00025## 45 4% 3% 7% 22 ##STR00026## 22 20% 4% 24% 23
##STR00027## 47 4% -84% -80% 24 ##STR00028## 11 20% 1% 21% 25
##STR00029## 17 10% -3% 7% 26 ##STR00030## 32 28% 9% 37% 27
##STR00031## 35 -17% -23% -40% 28 ##STR00032## 18 0% -8% -8% 29
##STR00033## 48 19% 11% 30% 30 ##STR00034## 17 -10% -34% -44% 31
##STR00035## 43 9% 9% 18%
[0078] Tables IA, IB and IC summarize the results obtained in the
experiments described above. Table IA shows the results obtained
with compounds encompassed by Formula I (i.e. the invention).
Tables IB and IC are included for comparative purposes and include
compounds not described by Formula I. Table IB shows diamides
structurally related to those of Formula I. Table IC shows the
results obtained with known ACAT inhibitors, not structurally
related to the diamides, but having an IC.sub.50 of 50 nm, or less,
when measured in the ACAT assay described supra.
[0079] To expedite this comparison a common format was used in the
three tables. Each compound was assigned an arbitrary compound
number, which is shown in the far left column (i.e. column #1). The
second column shows the structure of the compound tested and the
third column shows its potency as an ACAT inhibitor in the assay
supra.
[0080] Columns 4 through 6 shows the results the compounds had on
the secretion of sebum. The results are expressed as the difference
from the control. A positive number reflects a decrease in the
production of the sebum component being measured, i.e. cholesterol
ester (CE) or wax ester (WE). A negative number indicates that the
compound increased the production of CE or WE.
[0081] Column 4 shows the compounds ability to reduce the amount of
cholesterol ester in the sebum sample. Inhibition of cholesterol
ester is important because ACAT is responsible for the conversion
of cholesterol to cholesterol ester. These results reflect the
compounds ability, or lack thereof, to module ACAT in the target
tissue (hamster sebaceous glands).
[0082] Column 5 shows the effect the compound had on the generation
of wax ester. Wax esters are specific markers of the sebaceous
glands and are not appreciably detected in any other layer of the
skin. Reduction of wax ester reflects a decrease in sebum
secretion. Columns 6 is a summation of the results expressed in
columns 4 and 5 (and is included to further elucidate relative
differences in activity).
[0083] As shown in Table IA, the diamides of Formula I
significantly decreased the secretion of cholesterol ester,
indicating that ACAT was being inhibited in the target tissue. Wax
ester was also decreased indicating that total sebum secretion was
diminished when compared to the control. For example compound #1
decreased CE by 95% and WE by 80%. Compounds 2 and 3 produced
comparable results.
[0084] A comparison with Table IB shows significant differences.
Despite the structural similarity, these diamides had significantly
less impact on CE and WE secretion.
[0085] A comparison with the compounds of Table IC is also
illustrative. These compounds are all potent ACAT inhibitors. All
had IC.sub.50's of 50 nm, or less. Despite this potency, as a
group, they had significantly less effect on sebum secretion than
the compound of Formula I. Such results were unexpected.
* * * * *