U.S. patent application number 11/286825 was filed with the patent office on 2006-06-15 for methods and related compositions for reduction of fat and skin tightening.
Invention is credited to Michael S. Kolodney, Adam M. Rotunda.
Application Number | 20060127468 11/286825 |
Document ID | / |
Family ID | 46323236 |
Filed Date | 2006-06-15 |
United States Patent
Application |
20060127468 |
Kind Code |
A1 |
Kolodney; Michael S. ; et
al. |
June 15, 2006 |
Methods and related compositions for reduction of fat and skin
tightening
Abstract
Compositions and methods useful in the reduction of localized
fat deposits and tightening of loose skin in subjects in need
thereof using pharmacologically active detergents are disclosed.
The pharmacologically active detergent compositions can
additionally include anti-inflammatory agents, analgesics,
dispersion or anti-dispersion agents and pharmaceutically
acceptable excipients. The pharmacologically active detergent
compositions are useful for treating localized accumulations of fat
including, for example, lower eyelid fat herniation, lipodystrophy
and fat deposits associated with cellulite and do not require
surgical procedures such as liposuction.
Inventors: |
Kolodney; Michael S.; (Santa
Monica, CA) ; Rotunda; Adam M.; (Los Angeles,
CA) |
Correspondence
Address: |
PRESTON GATES & ELLIS LLP
1900 MAIN STREET, SUITE 600
IRVINE
CA
92614-7319
US
|
Family ID: |
46323236 |
Appl. No.: |
11/286825 |
Filed: |
November 23, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11134727 |
May 19, 2005 |
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11286825 |
Nov 23, 2005 |
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11054171 |
Feb 8, 2005 |
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11134727 |
May 19, 2005 |
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60572879 |
May 19, 2004 |
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Current U.S.
Class: |
424/450 ;
514/171; 514/78 |
Current CPC
Class: |
A61K 8/63 20130101; A61K
31/575 20130101; A61Q 19/06 20130101; A61K 31/185 20130101; A61K
31/685 20130101; A61K 31/185 20130101; A61K 47/24 20130101; A61K
45/06 20130101; A61K 2800/91 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 35/35 20130101; A61K
31/685 20130101; A61K 31/56 20130101; A61K 9/0019 20130101; A61K
47/28 20130101; A61K 31/56 20130101; A61Q 19/08 20130101; A61P 3/04
20180101 |
Class at
Publication: |
424/450 ;
514/078; 514/171 |
International
Class: |
A61K 31/685 20060101
A61K031/685; A61K 31/56 20060101 A61K031/56; A61K 9/127 20060101
A61K009/127 |
Claims
1. A pharmaceutical solution for subcutaneous injection comprising:
a therapeutically effective amount of a bile acid or salt thereof;
a pharmaceutical excipient; and optionally a lipid, wherein the
ratio of the lipid and bile acid or bile salt is less than 1% w/w
and wherein the solution does not include lipase or colipase.
2. The solution of claim 1 wherein said solution further comprises
a second therapeutic agent selected from the group consisting of:
anti-microbial agents, vasoconstrictors, anti-thrombotic agents,
anti-coagulation agents, suds-depressants, anti-inflammatory
agents, analgesics, dispersion agents, anti-dispersion agents,
penetration enhancers, steroids, tranquilizers, muscle relaxants,
and anti-diarrhea agents.
3. The solution of claim 1 wherein said solution is in a container
that contains up to 500 mL of solution.
4. The solution of claim 1 wherein said bile acid is selected from
the group consisting of: deoxycholic acid, cholic acid,
chenodeoxycholic acid, 7-alpha-dehydroxylate chenodeoxycholic acid,
lithocholic acid, ursodeoxycholic acid, dihydroxytaurin acid,
trihydroxytaurine acid, and glycine conjugates of any of the
above.
5. The solution of claim 1 wherein said bile salt comprises a
cation selected from the group consisting of sodium (Na.sup.+),
potassium (K.sup.+), lithium (Li.sup.+), magnesium (Mg.sup.2+),
calcium (Ca.sup.2+), barium (Ba.sup.2+), strontium (Sr.sup.2+), and
ammonium (NH.sup.4+).
6. The solution of claim 1 wherein said bile salt comprises an
alkali metal or an alkaline earth metal.
7. The solution of claim 6 wherein said alkali metal is selected
from the group consisting of alkali metal is sodium (Na.sup.+),
potassium (K.sup.+), and lithium (Li.sup.+).
8. The solution of claim 6 wherein said alkaline earth metal is
selected from the group consisting of magnesium (Mg.sup.2+),
calcium (Ca.sup.2+), barium (Ba.sup.2+), and strontium
(Sr.sup.2+).
9. The solution of claim 1 wherein said bile acid salt is sodium
deoxycholate.
10. The solution of claim 1 comprising up to 5% w/w lipids.
11. The solution of claim 1 wherein said lipid is
phosphatidylcholine.
12. The solution of claim 1 wherein said bile acid or bile salt is
at a concentration of about 0.001% w/w to about 10% w/w.
13. The solution of claim 1 wherein said bile acid or salt thereof
is at a concentration of about 0.01% w/w to about 5% w/w.
14. The solution of claim 1 wherein said solution is aqueous.
15. The solution of claim 1 wherein said therapeutically effective
amount is effective for fat dissolution or skin tightening.
16. A method for reducing the appearance of a skin condition in a
skin region of a subject comprising: administering locally to said
skin region a composition comprising: (i) a skin-tightening
effective amount of at least one bile acid or bile salt, (ii) a
pharmaceutical excipient, and (iii) optionally a lipid.
17. The method of claim 16 wherein said administering step involves
delivering said composition via a subcutaneous or transdermal
injection.
18. The method of claim 16 wherein said administering step involves
delivering said composition via a dermal patch, a pump, or
subdermal depot.
19. The method of claim 16 wherein said administering step involves
delivering said composition topically or subcutaneously.
20. The method of claim 16 wherein said skin condition is selected
from the group consisting of: loose skin, skin aging,
irregularities of the skin, and wrinkles.
21. The method of claim 16 wherein said region of skin is under
eye, under chin, under arm, buttock, cheek, brow, calf, back,
thigh, ankle, or stomach.
22. The method of claim 16 wherein said bile acid is deoxycholic
acid.
23. The method of claim 16 wherein said bile salt is sodium
deoxycholate.
24. The method of claim 16 wherein said bile salt is sodium
deoxycholate and is up to 5% w/w.
25. The method of claim 16 wherein the ratio of said lipid and said
bile acid or bile salt is up to 1% w/w.
26. The method of claim 16 wherein said lipid is
phosphatidylcholine.
27. The method of claim 16 wherein said lipid is
phosphatidylcholine and is up to 5% w/w.
28. The method of claim 16 wherein said composition is an aqueous
solution.
29. The method of claim 18 wherein said aqueous solution has total
volume up to 500 mL.
30. A method for reducing a subcutaneous fat deposit in a subject
comprising: administering locally to said subcutaneous fat deposit
a composition comprising: (i) a fat-dissolving effective amount of
one or more bile acids or bile salts; (ii) a pharmaceutical or
veterinary excipient; and (iii) optionally a lipid, wherein the
ratio of the lipid and bile acid or bile salt is up to 1% w/w and
wherein the composition does not include lipase or colipase.
31. The method of claim 30 wherein said condition is selected from
the group consisting of obesity, fat redistribution syndrome,
eyelid fat herniation, lipomas, Dercum's disease, lipodystrophy,
buffalo hump lipodystrophy, dorsocervical fat, visceral adiposity,
breast enlargement, hyperadiposity, diffused body fat around trunk
and arms, and fat deposits associated with cellulite.
32. The method of claim 30 wherein said bile acid is selected from
the group consisting of deoxycholic acid, cholic acid,
chenodeoxycholic acid, 7-alpha-dehydroxylate chenodeoxycholic acid,
lithocholic acid, ursodeoxycholic acid, dihydroxytaurin acid,
trihydroxytaurine acid, and glycine conjugates of any of the
above.
33. The method of claim 30 wherein said bile salt comprises a
cation selected from the group consisting of sodium (Na.sup.+),
potassium (K.sup.+), lithium (Li.sup.+), magnesium (Mg.sup.2+),
calcium (Ca.sup.2+), barium (Ba.sup.2+), strontium (Sr.sup.2+), and
ammonium (NH.sup.+).
34. The method of claim 30 wherein said bile salt comprises an
alkali metal or an alkaline earth metal.
35. The method of claim 34 wherein said alkali metal is sodium
(Na.sup.+), potassium (K.sup.+), or lithium (Li.sup.+).
36. The method of claim 34 wherein said alkaline earth metal is
magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium (Ba.sup.2+), or
strontium (Sr.sup.2+).
37. The method of claim 30 wherein said bile salt is sodium
deoxycholate.
38. The method of claim 30 wherein said method does not include
performing surgery on said subject.
39. The method of claim 30 wherein said composition is administered
locally under eye, under chin, under arm, buttock, calf, back,
thigh, or stomach of said subject.
40. The method of claim 30 wherein said administering is by a
subcutaneous or transdermal injection.
41. The method of claim 30 wherein said subject is a human, cat,
dog or horse.
42. The method of claim 30 wherein said subject is human HIV
patient
43. The method of claim 30 wherein said composition includes up to
5% w/w lipids.
44. The method of claim 30 wherein said lipid is
phosphatidylcholine and wherein said composition includes up to 5%
w/w of said lipid.
45. The method of claim 30 wherein said composition comprises about
0.001% w/w to about 10% w/w of said bile acid or bile salt.
46. The method of claim 30 wherein said composition comprises
between about 0.01% w/w and about 5% w/w of said bile acid or bile
salt.
47. The method of claim 30 wherein said composition is in
solution.
48. The method of claim 47 wherein said solution is aqueous
49. A syringe loadable container comprising: a fat-dissolving or
skin-tightening effective amount of a bile acid or a bile salt; a
pharmaceutical or veterinary excipient; and optionally a lipid
wherein the ratio of said bile acid or bile salt to said lipid is
greater than 1% w/w and wherein the solution does not contain
lipase or colipase.
50. The syringe loadable container of claim 47 wherein said bile
salt is sodium deoxycholate.
51. The syringe loadable container of claim 47 further comprising
an organic solvent.
52. The syringe loadable container of claim 47 comprising up to 10
g of said bile acid or bile salt.
Description
CROSS-REFERENCE
[0001] This application is a continuation-in-part application of
Ser. No. 11/134,727, filed May, 19, 2005 which is a
continuation-in-part application of Ser. No. 11/054,171, filed Feb.
8, 2005, which claims priority to U.S. Provisional Application Ser.
No. 60/572,879 filed May 19, 2004, which are incorporated herein by
reference in its entirety.
INCORPORATION OF REFERENCE
[0002] All references disclosed herein are incorporated in their
entirety by reference for any and all purposes.
BACKGROUND OF THE INVENTION
[0003] Surgical and non-surgical procedures for improving
appearance have increased in prevalence as populations age and gain
weight. Liposuction is a popular cosmetic surgery procedure and
involves the surgical removal of fat deposits using suction and
optionally assisted by solutions to assist in fat removal.
Liposuction is a surgical procedure that removes fat through an
incision in the skin through which a cannula is inserted. The
cannula is connected to a suction source and the unwanted fat is
aspirated through the cannula and discarded. Liposuction is
performed under general or local anesthesia, depending on the
amount and location of the fat to be removed. However, liposuction
and other surgical methods of fat removal are associated with
significant adverse events including temporary bruising, swelling,
numbness, soreness and burning sensation, risk of infection,
pigmentation changes, the formation of fat clots or blood clots
which can migrate to the lungs and cause death, excessive fluid
loss, which can lead to shock or fluid accumulation that must be
drained, friction burns or other damage to the skin or nerves or
perforation injury to the vital organs. Additionally, liposuction
requires a recovery time of one to two weeks wherein the patient
cannot work or perform certain daily activities. Moreover, because
surgical procedures such as liposuction require local and
occasionally general anesthesia, significant anesthesia-related
risks are associated with surgical fat removal. Furthermore,
liposuction and other drastic weight loss methods may result in
loose skin.
[0004] Therefore it would be desirable to have compositions and
methods for removing localized fat accumulations that does not
require surgery or prolonged recovery time as well as for
tightening loose skin.
SUMMARY OF THE INVENTION
[0005] The present invention provides compositions, methods, and
kits for reducing subcutaneous fat deposits as well as tightening
loose skin.
[0006] In one aspect, the compositions herein are in a solution.
Preferably the solution is aqueous.
[0007] In one embodiment, the present invention relates to a
solution for subcutaneous injection comprising: (i) a
therapeutically effective amount of one or more pharmacologically
active detergents, or bile acid(s) and/or bile salt(s), or
deoxycholic acid or a salt thereof, or sodium deoxycholate; (ii) a
pharmaceutical, veterinary, or cosmetic excipient; and (iii)
optionally a lipid, wherein the ratio of the lipid and bile acid or
bile salt is less than 1% w/w and wherein the solution does not
include lipase or colipase.
[0008] In some embodiments, the above solution can further comprise
a second therapeutic agent selected from the group consisting of:
anti-microbial agents, vasoconstrictors, anti-thrombotic agents,
anti-coagulation agents, suds-depressants, anti-inflammatory
agents, analgesics, dispersion agents, anti-dispersion agents,
penetration enhancers, steroids, tranquilizers, muscle relaxants,
and anti-diarrhea agents.
[0009] In some embodiments, a solution is in a container that
contains up to 500 mL of solution. Such container can be a syringe
or syringe-loadable container.
[0010] In some embodiments, a solution comprises a bile acid
selected from the group consisting of: deoxycholic acid, cholic
acid, chenodeoxycholic acid, 7-alpha-dehydroxylate chenodeoxycholic
acid, lithocholic acid, ursodeoxycholic acid, dihydroxytaurin acid,
trihydroxytaurine acid, and glycine conjugates of any of the
above.
[0011] In some embodiments, a solution comprises a bile salt
wherein the salt comprises a cation selected from the group
consisting of sodium (Na.sup.+), potassium (K.sup.+), lithium
(Li.sup.+), magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium
(Ba.sup.2+), strontium (Sr.sup.2+), and ammonium (NH.sup.4+).
Preferably, when the above solution comprises a bile salt, the bile
salt is sodium deoxycholate.
[0012] In some embodiments, a solution comprises a bile salt that
includes an alkali metal or an alkaline earth metal. Preferably an
alkali metal is selected from the group consisting of alkali metal
is sodium (Na.sup.+), potassium (K.sup.+), and lithium (Li.sup.+).
Preferably, an alkaline earth metal is selected from the group
consisting of magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium
(Ba.sup.2+), and strontium (Sr.sup.2+).
[0013] The compositions (e.g., solutions) include a therapeutically
effective amount of the pharmacological detergents (e.g., bile
acid(s) and/or bile salt(s). Such therapeutically effective amounts
are effective to reduce a subcutaneous fat deposit or tighten a
region of loose skin.
[0014] The bile acid(s) or bile salt(s) in a solution of the
invention can be at a concentration of about 0.001 to 10, 0.01 to
5, or 0.1 to 2% w/w, w/v, or v/v. Preferably, the bile acid(s) or
bile salt(s) in the above solution can be at a concentration of
about 0.1-5% w/w or more preferably about 1% w/w.
[0015] In some embodiments, the solutions herein include no lipids,
phospholipids, or phosphatidylcholine. In some embodiments, the
solutions herein include up to 5% w/w, w/v, or v/v lipids,
phospholipids, or phosphatidylcholine.
[0016] In one aspect, the present invention relates to methods for
reducing the appearance of a skin condition in a skin region of a
subject. Such methods comprise the step of: administering locally
to said skin region a composition comprising: (i) a skin-tightening
effective amount of one or more pharmacologically active
detergents, or bile acid(s) and/or bile salt(s), or deoxycholic
acid or a salt thereof, or sodium deoxycholate, (ii) a
pharmaceutical, veterinary, or cosmetic excipient, and (iii)
optionally a lipid.
[0017] In some embodiments, the administering step involves
delivering the compositions herein via a subcutaneous or
transdermal injection.
[0018] In some embodiments, the administering step involves
delivering the compositions herein via a dermal patch, a pump, or
subdermal depot.
[0019] In some embodiments, the administering step involves
delivering the compositions herein topically or subcutaneously.
[0020] In some embodiments, the skin condition being treated or
ameliorated is selected from the group consisting of: loose skin,
skin aging, irregularities of the skin, and wrinkles.
[0021] In some embodiments, the region of skin being treated is
under eye, under chin, under arm, buttock, cheek, brow, calf, back,
thigh, ankle, or stomach.
[0022] In some embodiments, the compositions used for reducing the
appearance of a skin condition in a skin region are formulation
into a skin tightening solution.
[0023] Such skin tightening solution can further comprise a second
therapeutic agent selected from the group consisting of:
anti-microbial agents, vasoconstrictors, anti-thrombotic agents,
anti-coagulation agents, suds-depressants, anti-inflammatory
agents, analgesics, dispersion agents, anti-dispersion agents,
penetration enhancers, steroids, tranquilizers, muscle relaxants,
and anti-diarrhea agents.
[0024] In some embodiments, the skin tightening solution is in a
container that contains up to 500 mL of solution. Such container
can be a syringe or syringe-loadable container.
[0025] In some embodiments, the skin tightening solution comprises
a bile acid selected from the group consisting of: deoxycholic
acid, cholic acid, chenodeoxycholic acid, 7-alpha-dehydroxylate
chenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid,
dihydroxytaurin acid, trihydroxytaurine acid, and glycine
conjugates of any of the above.
[0026] In some embodiments, a solution comprises a bile salt
wherein the salt comprises a cation selected from the group
consisting of sodium (Na.sup.+), potassium (K.sup.+), lithium
(Li.sup.+), magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium
(Ba.sup.2+), strontium (Sr.sup.2+), and ammonium (NH.sup.+).
Preferably, when the above solution comprises a bile salt, the bile
salt is sodium deoxycholate.
[0027] In some embodiments, the skin tightening solution comprises
a bile salt that includes an alkali metal or an alkaline earth
metal. Preferably an alkali metal is selected from the group
consisting of alkali metal is sodium (Na.sup.+), potassium
(K.sup.+), and lithium (Li.sup.+). Preferably, an alkaline earth
metal is selected from the group consisting of magnesium
(Mg.sup.2+), calcium (Ca.sup.2+), barium (Ba.sup.2+), and strontium
(Sr.sup.2+).
[0028] In some embodiments, the skin tightening solution includes a
therapeutically effective amount of the pharmacological detergents
(e.g., bile acid(s) and/or bile salt(s). Such therapeutically
effective amount is effective to tighten a loose skin region.
[0029] In some embodiments, the skin tightening solution comprises
one or more pharmacologically active detergents (e.g., bile acid(s)
and/or bile salt(s), such as sodium deoxycholate) at a
concentration of about 0.001 to 10, 0.01 to 5, or 0.1 to 2% w/w,
w/v, or v/v. Preferably, the one or more pharmacologically active
detergents in the skin tightening solution is at a concentration of
about 0.1-5% w/w, or more preferably about 1% w/w.
[0030] In some embodiments, the skin tightening solution comprises
up to 100, 50, 20, 10, 5, 2, 1, 0.5, 0.2, 0.05, 0.02, or 0.01 grams
of the one or more detergents, bile acids and/or bile salts,
deoxycholic acid or salts thereof or sodium deoxycholate.
[0031] In some embodiments, the skin tightening solution includes
no lipids, phospholipids, or phosphatidylcholine. In some
embodiments, the solutions herein include up to 5% w/w, w/v, or v/v
lipids, phospholipids, or phosphatidylcholine.
[0032] In one aspect, the present invention relates to methods for
reducing a subcutaneous fat deposit in a subject. Such methods
comprise the step of administering locally to a subcutaneous fat
deposit in the subject a composition comprising: (i) a
fat-dissolving effective amount of one or more pharmacologically
active detergents, or bile acid(s) and/or bile salt(s), or
deoxycholic acid or a salt thereof, or sodium deoxycholate; (ii) a
pharmaceutical, veterinary, or cosmetic excipient; and (iii)
optionally a lipid, wherein the ratio of the lipid and bile acid or
bile salt is up to 1% w/w and wherein the composition does not
include lipase or colipase.
[0033] In some embodiments, the fat deposit is associated with a
condition selected from the group consisting of obesity, fat
redistribution syndrome, eyelid fat herniation, lipomas, Dercum's
disease, lipodystrophy, buffalo hump lipodystrophy, dorsocervical
fat, visceral adiposity, breast enlargement, hyperadiposity,
diffused body fat around trunk and arms, and fat deposits
associated with cellulite.
[0034] In some embodiments, the detergent comprises a bile acid
selected from the group consisting of deoxycholic acid, cholic
acid, chenodeoxycholic acid, 7-alpha-dehydroxylate chenodeoxycholic
acid, lithocholic acid, ursodeoxycholic acid, dihydroxytaurin acid,
trihydroxytaurine acid, and glycine conjugates of any of the
above.
[0035] In some embodiments, the detergent comprises a bile salt
that includes a cation selected from the group consisting of sodium
(Na.sup.+), potassium (K.sup.+), lithium (Li.sup.+), magnesium
(Mg.sup.2+), calcium (Ca.sup.2+), barium (Ba.sup.2+), strontium
(Sr.sup.2+), and ammonium (NH.sup.4+).
[0036] In some embodiments, the detergent comprises a bile salt
with a cation that is an alkali metal or an alkaline earth metal.
Preferably, the alkali metal is sodium (Na.sup.+), potassium
(K.sup.+), or lithium (Li.sup.+) and the alkaline earth metal is
magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium (Ba.sup.2+), or
strontium (Sr.sup.2+). More preferably, the bile salt is sodium
deoxycholate.
[0037] In some embodiments, the above method does not include
performing surgery on said subject.
[0038] In some embodiments, the administration step involves
administering locally (e.g., subcutaneously or subdermally) to a
region under eye, under chin, under arm, buttock, calf, back,
thigh, or stomach of said subject. The administration can be made
by a subcutaneous or transdermal injection.
[0039] The subject being treated by the compositions herein can be
a human or a domesticated animal such as a cat, dog, or horse. In
some embodiments, the subject being treated is a human HIV patient.
Such patient can be undergoing a HIV protease treatment or
experiencing or susceptible to experiencing lipodystriphy.
[0040] In some embodiments, the composition (e.g., solution or
aqueous solution) being administered includes up to 5% w/w, w/v or
v/v lipids, phospholipids, or phosphatidylcholine. Preferably, the
composition (e.g., solution or aqueous solution) being administered
includes up to 5% w/w phosphatidylcholine.
[0041] In some embodiments, the composition (e.g., solution or
aqueous solution) being administered comprises 0.001 to 10, 0.01 to
5, or 0.1 to 2% w/w, w/v, or v/v detergent(s) or bile acid(s)
and/or bile salt(s). More preferably a composition (e.g., solution
herein) comprises about 0.1-5% w/w or more preferably about 1% w/w
bile salts such as sodium deoxycholate.
[0042] Preferably, the compositions once administered are not
removed from the subject. Furthermore, the compositions herein are
preferably administered without a surgical procedure (e.g.,
liposuction).
[0043] In some embodiments, the fat dissolving solution comprises
up to 100, 50, 20, 10, 5, 2, 1, 0.5, 0.2, 0.05, 0.02, or 0.01 grams
of the one or more detergents, bile acids and/or bile salts,
deoxycholic acid or salts thereof or sodium deoxycholate.
[0044] In one aspect, the present invention relates to a syringe
loadable container comprising: (i) a fat-dissolving or
skin-tightening effective amount of one or more pharmacologically
active detergents, or bile acid(s) and/or bile salts(s), or
deoxycholic acid or a salt thereof, or sodium deoxycholate; (ii) a
pharmaceutical, veterinary, or cosmetic excipient; and (iii)
optionally a lipid wherein the ratio of said bile acid or bile salt
to said lipid is greater than 1% w/w and wherein the solution does
not contain lipase or colipase.
[0045] Preferably, the one or more detergents herein comprise
sodium deoxycholate.
[0046] Preferably the container contains up to 500, 200, 100, 50,
20, 10, 5, 2, or 1 mL of a solution that is sterile.
[0047] In some embodiments, the solution comprises an organic
solvent, or more preferably benzyl alcohol.
[0048] In some embodiments, the solution comprises 0.001 to 10,
0.01 to 5, or 0.1 to 2% w/w, w/v, or v/v detergent(s) or bile
acid(s) and/or bile salt(s). More preferably the solution comprises
about 0.1-5% w/w or more preferably about 1% w/w bile salts such as
sodium deoxycholate.
[0049] In some embodiments, the solution comprises up to 100, 50,
20, 10, 5, 2, 1, 0.5, 0.2, 0.05, 0.02, or 0.01 grams of the one or
more detergents, bile acids and/or bile salts, deoxycholic acid or
salts thereof or sodium deoxycholate.
INCORPORATION BY REFERENCE
[0050] All publications and patent applications mentioned in this
specification are herein incorporated by reference to the same
extent as if each individual publication or patent application was
specifically and individually indicated to be incorporated by
reference.
BRIEF DESCRIPTION OF THE DRAWINGS
[0051] FIG. 1 depicts the molecular structure of (a)
phosphatidylcholine (b) sodium deoxycholate and (c) benzyl
alcohol.
[0052] FIG. 2 depicts the effects of phosphatidylcholine bile salt
formulation (PBF) (5.0% highly purified soy derived PC, 4.75%
sodium deoxycholate, and 0.9% benzyl alcohol, in sterile water) and
sodium deoxycholate alone on cultured cell viability according to
the teachings of the present invention: (a) MTS assay measuring
viability of keratinocytes exposed to the PBF and sodium
deoxycholate alone; (b) lactate dehydrogenase (LDH) assay measuring
LDH release by cells exposed to the PBF and sodium deoxycholate
alone.
[0053] FIG. 3 depicts the effects of PBF and sodium deoxycholate
alone on primary porcine fat tissue according to the teachings of
the present invention: (a) MTS assay producing purple pigment,
indicating living cells, in fat specimens treated with the PBS
buffer as negative control (-Cont), sodium deoxycholate alone (DC),
the PBF, and Triton detergent as positive control (+Cont); (b) A
comparison of fat cell viability between the different
treatments.
[0054] FIG. 4 depicts calcein fluorescence in fat specimens treated
with sodium deoxycholate alone (DC), PBF, Triton detergent as
positive control (+Cont), and PBS buffer as negative control
(-Cont) according to the teachings of the present invention.
[0055] FIG. 5 depicts light microscopy of porcine skin biopsies
after treatment revealing (a) control lipocytes and (b) lipocytes
after PBF injection (H&E, original magnification, .times.20);
(c) control lipocytes and (d) lipocytes after injection of sodium
deoxycholate alone (H&E, original magnification, .times.10);
(e) control muscle and (f) muscle after injection of
phosphatidylcholine alone (H&E, original magnification,
.times.10); (g) fat after injection with Empigen detergent
(H&E, original magnification, .times.20).
[0056] FIG. 6 depicts a lipoma removed from a patient two days
after injection with deoxycholate according to the teachings of the
present invention: (a) gross pathology and (b) histology (H&E,
original magnification, .times.20).
[0057] FIG. 7 depicts effects of sodium deoxycholate only and
sodium deoxycholate-1% phosphatidylcholine solutions on
melanocytes.
[0058] FIG. 8 depicts effects of sodium deoxycholate only and
sodium deoxycholate-1% phosphatidylcholine solutions on
adipocytes.
[0059] FIG. 9 depicts effects of addition of phosphatidylcholine to
4.75% sodium deoxycholate solutions on viable adipocytes.
[0060] FIG. 10 depicts inhibition of adipolysis by pre-incubation
with human lipoma fat.
[0061] FIG. 11 depicts a kit for reducing a subcutaneous fat
accumulation.
DETAILED DESCRIPTION OF THE INVENTION
[0062] The present invention addresses problems of localized fat
accumulation and loose skin in animals, such as humans. In one
aspect, the present invention provides compositions for reducing
fat deposits and tightening skin. Such compositions comprise,
consist essentially of, or consist of one or more pharmacologically
active detergents, more preferably bile acids or bile salts, more
preferably deoxycholic acid or a salt thereof, or more preferably
sodium deoxycholate. The amount of such detergent(s) in the
composition is an effective amount to dissolve or reduce a
subdermal fat deposit or to tighten loose skin. Such effective
amount will depend, in part, on the location of target site, size
of target site, length of treatment, etc. In some of the
embodiments, a composition includes at least 2, 3, 4, 5, 6, 7, 8,
9, or 10 pharmacologically active detergents.
[0063] Pharmacologically active and biologically compatible
detergents include, but are not limited to, lipophilic detergents
(whether ionic or non-ionic), hydrophilic detergents (whether ionic
or non-ionic), ionic detergents, non-ionic detergents, zwitterionic
detergents, glycerides, bile acids and bile salts.
[0064] Non-limiting examples of lipophilic detergents include,
inter alia, alcohols, polyoxyethylene alkylethers, fatty acids,
bile acids, glycerol fatty acid esters, acetylated glycerol fatty
acid esters, lower alcohol fatty acids esters, polyethylene glycol
fatty acid esters, polyethylene glycol glycerol fatty acid esters,
polypropylene glycol fatty acid esters, polyoxyethylene glycerides,
lactic acid derivatives of mono/diglycerides, propylene glycol
diglycerides, sorbitan fatty acid esters, polyoxyethylene sorbitan
fatty acid esters, polyoxyethylene-polyoxypropylene block
copolymers, transesterified vegetable oils, sterols, sterol
derivatives, sugar esters, sugar ethers, sucroglycerides,
polyoxyethylene vegetable oils, polyoxyethylene hydrogenated
vegetable oils, reaction mixtures of polyols and at least one
member of the group consisting of fatty acids, glycerides,
vegetable oils, hydrogenated vegetable oils, and sterols, and
mixtures thereof.
[0065] Non-limiting examples of non-ionic lipophilic detergents
include, inter alia, alkylglucosides, alkylmaltosides,
alkylthioglucosides, lauryl macrogolglycerides, polyoxyethylene
alkyl ethers, polyoxyethylene alkylphenols, polyethylene glycol
fatty acids esters, polyethylene glycol glycerol fatty acid esters,
polyoxyethylene sorbitan fatty acid esters,
polyoxyethylene-polyoxypropylene block copolymers, polyglycerol
fatty acid esters, polyoxyethylene glycerides, polyoxyethylene
sterols, derivatives, and analogues thereof, polyoxyethylene
vegetable oils, polyoxyethylene hydrogenated vegetable oils,
reaction mixtures of polyols and at least one member of the group
consisting of fatty acids, glycerides, vegetable oils, hydrogenated
vegetable oils, and sterols, tocopherol polyethylene glycol
succinates, sugar esters, sugar ethers, sucroglycerides, and
mixtures thereof.
[0066] Non-limiting examples of ionic hydrophilic detergents
include, inter alia, alkyl ammonium salts, bile acids and bile
salts, analogues, and derivatives thereof, fatty acid derivatives
of amino acids, carnitines, oligopeptides, and polypeptides,
glyceride derivatives of amino acids, oligopeptides, and
polypeptides, acyl lactylates, mono-, diacetylated tartaric acid
esters of mono-, diglycerides, succinoylated monoglycerides, citric
acid esters of mono-, diglycerides, alginate salts, propylene
glycol alginate, lecithins and hydrogenated lecithins, lysolecithin
and hydrogenated lysolecithins, lysophospholipids and derivatives
thereof, phospholipids and derivatives thereof, salts of
alkylsulphates, salts of fatty acids, sodium docusate, and mixtures
thereof.
[0067] Non-limiting examples of ionic detergents include, but not
limited to, cholate, sodium deoxycholate, sodium dodecylsulfate and
C-16 TAB. In preferred embodiment, a non-limiting example of an
ionic detergent useful in an embodiment of the present invention is
sodium deoxycholate.
[0068] Non-limiting examples of non-ionic detergents include, but
not limited to, Brij 35, n-alkyl PEO monoether such as,
polyoxylethylen(20)cetyl ether, Lubrol PX, Lubrol WX, nonidet P-40,
n-alkyl phenyl PEO such as,
octylphenolpoly(ethyleneglycolether)n10, and
octylphenolpoly(ethyleneglycolether)n7, tetramethylbutylphenyl PEO,
n-octylglucoside, octyl-thioglucopyranoside, tween-80 and tween-20,
and alkylaryl polyether alcohol (Triton X-100).
[0069] Non-limiting examples of zwitterionic detergents include,
but not limited to,
3-[(3-cholamidopropyl)dimthylammonio]propane-sulfonate (CHAPS),
N-tetradecyl-N,N-dimethyl-3-ammoniu-1-propanesulfonate, cholic acid
sulfobetaine, lauryldimethylbetaine (Empigen BB) and zwittergent
3-14.
[0070] Non-limiting examples of glycerides include, inter alia,
mono-, di- or tri-glycerides. Such triglycerides include, inter
alia, vegetable oils, fish oils, animal fats, hydrogenated
vegetable oils, partially hydrogenated vegetable oils, synthetic
triglycerides, modified triglycerides, fractionated triglycerides,
and mixtures thereof.
[0071] Non-limiting examples of bile acids include ursodeoxycholic
acids, cholic acid, glycolic acid, alcoholic acid, taurocholic
acid, deoxycholic acid, glycodeoxycholic acid, taurodeoxycholic
acid, chenodeoxycholic acid, glycochenodeoxychloic acid, and
taurochenodeoxycholic acid, ursocholic acid, 7-oxolithocholic acid,
lithocholic acid 3-sulfate, norcholic acid, bisnorcholic acid,
hyocholic acid, and hyodeoxycholic acid.
[0072] In preferred embodiments the compositions herein comprise,
consist essentially of, or consist of pharmaceutically acceptable
salts and esters of the detergents. Such salts and esters are meant
to be those salts and esters which are within the scope of sound
medical judgment, suitable for use in contact with the tissues of
humans and animals without undue toxicity, irritation, allergic
response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use. Preferably a
composition herein comprises, consists essentially of or consists
of a bile salt, or more preferably deoxycholate salt, or more
preferably sodium deoxycholate.
[0073] Among detergents, bile salts are preferred as they are
particularly potent solubilizers of lipid bilayer membranes. All
biologic cell membranes are composed of the same bilipid structure,
and are therefore subject to solubilization by detergents.
Solubilization of cell membranes by a detergent involves
distribution of the detergent between lipid bilayers,
destabilization of the bilayer, disintegration, and subsequent
formation of mixed micelles (composed of detergent and cell
membrane lipid). Bile salts, and other detergents, decrease surface
tension at the border of immiscible materials and allow the
breakdown of large aggregates into smaller and smaller particles.
In tissue, these agents dissolve cell membranes and cause cell
lysis. An inflammatory response is generated, causing the body to
remove the detergent solubilized material.
[0074] Bile salts have been used to improve the aqueous solubility
of phosphatidylcholine (PC) and more recently, medications like
amphotericin B, Taxol, and diazepam. Highly purified
phosphatidylcholine can be combined with the secondary bile salt
sodium deoxycholate, an anti-microbial, benzyl alcohol, and water
to form a stable, mixed micelle preparation that can be rapidly
sterilized and used for intravenous administration. Pharmaceutical
preparations of this mixture, known as Essentiale and Lipostabil,
are marketed in other countries for treatment of liver disease and
hyperlipidemia, respectively.
[0075] Bile salts may be formed from a bile acid in combination
with cations such as inorganic bases, ammonia, organic bases, basic
amino acids or the like. Examples of the inorganic bases include
alkali metal (e.g., Li.sup.+, Na.sup.+ and K.sup.+) and alkaline
earth metal (e.g., Mg.sup.2+, Ca.sup.2+, Sr.sup.2+, Ba.sup.2+).
Examples of the organic base include procaine,
2-phenylethylbenzylamine, dibenzylethylenediamine, etanolamine, di
etanolamine, tris(hydroxymethyl)aminomethane,
polyhydroxyalkylamine, and N-methyl glucosamine. Examples of the
basic amino acid include lysine, arginine, ornithine and histidine.
Example of other salts includes halogen ions. Non limiting examples
of the salts that can be combined with bile acids to form bile
salts include, but are not limited to, ammonium chloride, ammonium
sulphate, sodium chloride, sodium bromide, sodium iodide, sodium
fluoride, sodium citrate, sodium sulphate, sodium carbonate, sodium
bicarbonate, sodium acetate, sodium nitrate, sodium nitrite,
potassium acetate, potassium carbonate, potassium dichromate,
potassium chloride, potassium bromide, magnesium bromide, magnesium
chloride, potassium iodide, sodium fluoride, hydroxylamine
hydrochloride, sodium fluoride, sodium silicate, diammonium
phosphate, sodium thiocyanate, potassium thiocyanate, lithium
thiocyanate, sodium borohydride, calcium carbonate, barium
carbonate, sodium dihydrogen phosphate, lithium chloride, lithium
bromide, and lithium iodide. In some embodiments, the salt ion that
combines with the bile acid to form a bile salt is a cation. Non
limiting examples of cations include sodium (Na.sup.+), potassium
(K.sup.+), lithium (Li.sup.+), magnesium (Mg.sup.2+), calcium
(Ca.sup.2+), barium (Ba.sup.2+), strontium (Sr.sup.2+), and
ammonium (NH.sup.4+).
[0076] Examples of bile salts that can be formed from a combination
of a bile acid and a cation, include, but are not limited to,
sodium cholate, sodium deoxycholic, sodium cholic, sodium
chenodeoxycholic, sodium 7-alpha-dehydroxylate chenodeoxycholic,
sodium lithocholic, sodium ursodeoxycholic, potassium deoxychote,
potassium cholate, potassium chenodeoxycholate, potassium
7-alpha-dehydroxylate chenodeoxycholate, potassium lithocholate,
potassium ursodeoxycholate, lithium deoxycholate, lithium cholate,
lithium chenodeoxycholate, lithium 7-alpha-dehydroxylate
chenodeoxycholate, lithium lithocholate, lithium ursodeoxycholate,
magnesium deoxycholate, magnesium cholate, magnesium
chenodeoxycholate, magnesium 7-alpha-dehydroxylate
chenodeoxycholate, magnesium lithocholate, magnesium
ursodeoxycholate, ammonium cholate, ammonium deoxycholate, ammonium
cholate, ammonium chenodeoxycholate, ammonium 7-alpha-dehydroxylate
chenodeoxycholate, ammonium lithocholate, ammonium
ursodeoxycholate, dihydroxy- and trihydroxy- and taurine or glycine
conjugates of any of the above. Preferably a bile salt of the
invention is sodium deoxycholate. Any of the above bile salts can
be used in the compositions herein.
[0077] Other examples of bile salts include steroids having 1-3
hydroxyl groups and a five carbon atom side chain terminating in a
carboxyl group, which can be conjugated to glycine or taurine.
[0078] In some embodiments, a composition herein comprises,
consists essentially of, or consists of one or more esters of a
bile acid. Such esters include, but are not limited to, esters
formed by a combination of a bile acid with a hydrogen of a COOH
group optionally substituted C.sub.1-C.sub.6 alkyl, C.sub.2-C.sub.6
alkenyl, C.sub.3-C.sub.10 cycloalkyl, C.sub.3-C.sub.10
cycloalkyl(C.sub.1-C.sub.6)alkyl, optionally substituted
C.sub.6-C.sub.10 aryl, optionally substituted C.sub.7-C.sub.12
aralkyl, di(C.sub.6-C.sub.10)arylmethyl,
tri(C.sub.6-C.sub.10)arylmethyl, or a substituted silyl.
[0079] Examples of the optionally substituted C.sub.1-6 alkyl
include e.g., methyl, ethyl, n-propyl, n-butyl, t-butyl, n-pentyl,
and n-hexyl, each may be substituted with benzyloxy, C.sub.1-4
alkylsulfonyl (e.g., methanesulfonyl), trimethylsilyl, halogen
(e.g., F, Cl, and Br), acetyl, nitrobenzoyl, mesylbenzoyl,
phthalimide, succinoylimide, benzenesulfonyl, phenylthio,
di-C.sub.1-4 alkylamino (e.g., dimethylamino), pyridyl, C.sub.1-4
alkylsulfinyl (e.g., methanesulfinyl), cyano and the like. Such
substituted C.sub.1-6 alkyl include e.g., benzyloxymethyl,
2-methanesulfonylethyl, 2-trimethylsilylethyl,
2,2,2-trichloroethyl, 2-iodoethyl, acetylmethyl,
p-nitrobenzoylmethyl, p-mesylbenzoylmethyl, phthalimidemethyl,
succinoylimidemethyl, benzenesulfonylmethyl, phenylthiomethyl, and
1-dimethylaminoethyl. The above C.sub.2-6 alkenyl includes e.g.,
vinyl, aryl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl,
1,1-dimethylaryl, 3-methyl and 3-butenyl. The above C.sub.3-10
cycloalkyl includes e.g., cyclopropyl, cyclobutyl, cyclopentyl,
cyclohexyl, cycloheptyl, norbornyl, and adamantyl. The above
C.sub.3-10 cycloalkyl(C.sub.1-6)alkyl includes e.g.,
cyclopropylmethyl, cyclopentylmethyl, and cyclohexylmethyl. The
above C.sub.6-10 aryl includes e.g., phenyl, -naphthyl, 8-naphthyl,
and biphenyl, each may be substituted with nitro, halogen (e.g., F,
Cl, and Br) or the like, and such substituted aryl includes e.g.,
p-nitrophenyl and p-chlorophenyl. The above optionally substituted
C.sub.7-12 aralkyl includes e.g., benzyl, 1-phenylethyl,
2-phenylethyl, phenylpropyl and naphthylmethyl, each may be
substituted with nitro, C.sub.1-4 alkoxy (e.g., methoxy), C.sub.1-4
alkyl (e.g., methyl, ethyl), hydroxy or the like. Such substituted
group is exemplified by p-nitrobenzyl, p-methoxybenzyl (PMB), or
3,5-di-t-butyl-4-hydroxybenzyl. The above di(C.sub.6-10 aryl)methyl
includes benzhydryl and the C.sub.6-10 arylmethyl includes trityl,
and the substituted silyl includes trimethylsilyl and
tert-butyldimethylsilyl. Examples of the active ester include
organic phosphate esters (e.g., diethoxy phosphate ester and
diphenoxy phosphate ester), cyanomethyl ester, and the active
thioester includes esters formed with aromatic heterocyclicthio
compound (e.g., 2-pyridilthio ester).
[0080] Derivatives of bile acids can also be used as detergents.
Such derivatives include bile acid halides, bile acid azides, bile
acid anhydrides, mixed bile acid anhydride, bile acid amide, and
bile acid thioester. The bile acid halide includes bile acid
chloride such as deoxycholic chloride and bile acid bromide such as
deoxycholic acid bromide; the mixed bile acid anhydride includes
mixed monoalkylcarboxylic acid anhydride, mixed alphatic carboxylic
acid anhydride, aromatic carboxylic acid anhydride, organic
sulfonic acid anhydride, and wherein the active amide includes
amide formed with heterocyclic compound containing N atom.
[0081] The pharmacologically active detergents (including bile
acids and bile salts) are preferably micelle-forming compounds.
Micelles can significantly increase the solubility of hydrophobic
molecules not ordinarily soluble in water (e.g., the lipids that
comprise cell membranes) by burying their hydrophobic portions away
from aqueous solvent (e.g., water). In some embodiments, a
composition herein comprises homogenous micelles (micelles produced
by a single detergent). In some embodiments, a composition herein
comprises mixed micellar formations (micelles produced by two or
more compounds--one of which is a detergent).
[0082] In some embodiments, a composition comprising micelles with
an average size in the range of 10.sup.-9 m-10.sup.-5 m, 10.sup.-6,
5.times.10.sup.-9 m-10.sup.-6 m, 10.times.10.sup.-9 m-10.sup.-7, or
50.times.10.sup.-9-10.times.10.sup.-8 m. In some embodiments, an
average size of a micelle in a composition of the present invention
may be up to 10.sup.-5, 10.sup.-6, 10.sup.-7, 10.sup.-8, 10.sup.-9
m. In some embodiments, an average size of a micelle in a
composition of the present invention may be greater than 10.sup.-5,
10.sup.-6, 10.sup.-7, 10.sup.-8, 10.sup.-9 m. Moreover, the shape
of the micelle can vary and can be, for example, prolate, oblate or
spherical; spherical micelles are most typical.
[0083] Table 1 below illustrates several detergents contemplated by
the present invention, their monomeric molecular weight of these
detergents as monomers, and their critical micellar concentration
(CMCs), which is the minimum concentration at which the detergent
is predominantly in the form of micelles. TABLE-US-00001 TABLE 1
Micellar Molecular Molecular CMC in Weight Weight H2O Detergent
Name (AMU) (AMU) (M) Anionic Cholate 430 4300 1.4 .times. 10-2
Deoxycholate 415-432 4200 5 .times. 10-3 Sodium dodecyl sulfate 288
18000 8.3 .times. 10-3 Cationic C16-TAB 365 62000 1 .times. 10-3
Amphoteric (Zwiterionic) Cholic acid-sulfobetaine 615 6150 4
.times. 10-3 Cholic acid-sulfobetaine 631 6940 8 .times. 10-3
Lysophophatidylcholine 495 92000 7 .times. 10-6 Zwitergent 3-14 364
30000 3 .times. 10-4 Non-Ionic Brij 35 1225 49000 9 .times. 10-5
polyoxylethylen(20)cetyl ether 1120 82000 7.7 .times. 10-5 Lubrol
PX 582 64000 1 .times. 10-4 Nonidet P-40 603 90000 3 .times. 10-4
Octylphenolpoly(ethylene- 647 90000 0.2 .times. 10-3
glycolether)n10 Octylphenolpoly(ethylene- 515 0.2 .times. 10-3
glycolether)n7 n-Octylglucoside 292 8000 14.5 .times. 10-3
Octyl-thioglucopyranoside 308 9 .times. 10-3 Tween-80 1310 76000
1.2 .times. 10-5 Tween-20 1228 6.0 .times. 10-5
[0084] In some embodiments, the concentration of the one or more
pharmacologically active detergents (e.g., bile acids or bile
salts) in a composition is such that it is at approximately the CMC
concentration (i.e., +/-5 mM), or above the CMC level (e.g., more
than 1, 5, 10 15, 20, 25, 30, 35, 40, 50, 55, 60, 65, 70, 75, 80,
85, 90, 99, 150, 200, 400, 800, 1600, 3200, 6800, 13,600, 27,200,
or 54,400%, above the CMC concentration level).
[0085] In some embodiments, at least 10, 15, 20, 25, 30, 35, 40,
45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 99% of the
detergents (e.g., bile acids and bile salts), in the compositions
are in micellar formation. In other embodiments, up to 90, 80, 70,
60, 50, 40, 30, 20, 10, or 5% of the detergents (e.g., bile acid(s)
and/or bile salt(s)), in the compositions are in micellar
formation. In other embodiments, about 10-90, 20-80, 30-70, 40-60,
or about 50% of the detergents (e.g., bile acids and bile salts),
of the compositions are in micellar formation.
[0086] In some embodiments, an average molecular weight of a
micelle in a composition of the present invention may be up to
100,000, 50,000, 40,000, 30,000, 20,000, 10,000, 9,000, 8,000,
7,000, 6,000, 5,000, 4,000, 3,000, 2,000, 1,000, or 500 Daltons
(D). In some embodiments, an average molecular weight of a micelle
in a composition of the present invention may be greater than 500,
1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, 5,000,
5,500, 6,000, 6,500, 7,000, 7,500, 8,000, 8,500, 9,000, 9,500,
10,000, or 15,000 D. In some embodiments, an average molecular
weight of a micelle in a composition of the present invention may
be in the range of 100-20,000, 1,000-10,000, 2,000-1,000, or
3,000-5,000 D.
[0087] In any of the embodiments herein the concentration of the
one or more pharmacologically active detergents (e.g., bile acid(s)
and/or bile salt(s) or more preferably sodium deoxycholate) can be
up to 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4,
3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05,
0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005, 0.004,
0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005,
0.0004, 0.0003, 0.0002, or 0.0001% w/w, w/v or v/v. Preferably, a
composition herein comprises a bile salt, such as sodium
deoxycholate, wherein the concentration of the bile salt is up to
5, 4, 3, 2 or 1% w/w.
[0088] In any of the embodiments herein the concentration of the
one or more pharmacologically active detergents (e.g., bile acid(s)
and/or bile salt(s) or more preferably sodium deoxycholate) is
greater than 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006,
0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006,
0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07,
0.08, 0.09, 0.10, 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.80, 0.90,
1.00, 1.25, 1.50, 1.75, 2.00, 2.25, 2.50, 2.75, 3.00, 3.25, 3.50,
3.75, 4.00, 4.25, 4.50, 4.75, 5.00, 5.25, 5.50, 5.75, 6.00, 6.25,
6.50, 6.75, 7.00, 7.25, 7.50, 7.75, 8.00, 8.25, 8.50, 8.75, 9.00,
9.25, 9.50, 9.75, or 10.00% w/w, w/v, or v/v. Preferably, a
composition herein comprises one or more bile salts, such as sodium
deoxycholate at a concentration of more than 0.0001, 0.001, 0.01 or
0.1% w/w.
[0089] In any of the embodiments herein, the concentration of the
pharmacologically active detergent(s) (e.g., bile acid(s) and/or
bile salt(s)) is approximately in the range 0.0001-50, 0.00140,
0.01-30, 0.02-29, 0.03-28, 0.04-27, 0.05-26, 0.06-25, 0.07-24,
0.08-23, 0.09-22, 0.1-21, 0.2-20, 0.3-19, 0.4-18, 0.5-17, 0.6-16,
0.7-15, 0.8-14, 0.9-12, or approximately in the range of 1-10% w/w,
w/v or v/v. Preferably, the concentration of the one or more of the
pharmacologically active detergents (e.g., bile acids or bile
salts) in a composition of the invention in the range from
approximately 0.005-15%, 0.01-10%, 0.05-15%, 0.10-1%, 0.1-10% or
0.5-1%, 0.5-5% w/w, w/v or v/v. It is understood that the final
concentration is dependent on many factors known to persons skilled
in the art including, but not limited to, location and size of the
target site.
[0090] In some embodiments, a composition contains up to 10, 9.5,
9.0, 8.5, 8.0, 7.5, 7.0, 6.5, 6.0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0,
2.5, 2.0, 1.5, 1.0, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6,
0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.09, 0.08,
0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007,
0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007,
0.0006, 0.0005, 0.0004, 0.0003, 0.0002, or 0.0001 grams of the one
or more pharmacologically active detergent(s) (e.g., bile acid(s)
and/or bile salt(s)), or more preferably up to 5, 4, 3, 2, 1, 0.5,
0.4, 0.3, 0.2 or 0.1 grams of a bile salt such as sodium
deoxycholate.
[0091] In some embodiments, a composition contains more than
0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008,
0.0009, 0.001, 0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045,
0.005, 0.0055, 0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009,
0.0095, 0.01, 0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05,
0.055, 0.06, 0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.10,
0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65,
0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5,
4.0 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10.0
grams of the one or more pharmacologically active detergents (e.g.,
bile acids or bile salts), or more preferably, more than 0.01,
0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08 or 0.09 grams.
[0092] In some embodiments, a composition herein contains between
0.0001-10, 0.0005-5, 0.001-8, 0.005-7, 0.01-6, 0.05-5, 0.1-4,
0.5-4, or 1-3 grams of the one or more pharmacologically active
detergent(s) (e.g., bile acid(s) and/or bile salt(s)), or more
preferably, between 0.001-10, 0.002-9, 0.003-8, 0.004-7, 0.005-6,
0.006-5, 0.0074, 0.008-3, 0.009-2, 0.01-1, 0.02-0.5, 0.03-0.4,
0.04-0.3, 0.05-0.2 or 0.06-0.1 grams of the one or more detergents
(e.g. sodium deoxycholate).
[0093] In some embodiments herein, a composition includes one or
more lipids, phospholipids, or phosphatidylcholine. Preferably, the
amount of lipids, phospholipids, or phosphatidylcholine in a
composition is at a concentration up to 50, 40, 30, 20, 15, 10, 9,
8, 7, 6, 5, 4, 3, 2, 1, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6,
0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.09, 0.08,
0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007,
0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007,
0.0006, 0.0005, 0.0004, 0.0003, 0.0002, or 0.0001% (w/w, w/v, or
v/v). In preferred embodiments, the amount of lipids,
phospholipids, or phosphatidylcholine in a composition is up to 5%
(w/w, w/v, or v/v). For example, the present invention contemplates
a composition comprising one or more detergents, preferably bile
salts, or more preferably sodium deoxycholate wherein the
composition includes up to 5, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3,
4.2, 4.1, 4.0, 3.0, 2.0, or 1.0% lipids or phospholipids or
phosphatidylcholine.
[0094] In some embodiments, the ratio between the detergent(s) and
lipids, phospholipids, or phosphatidylcholine is such that there is
more detergents by mass than lipids, phospholipids or
phosphatidylcholine respectively. For example, in some embodiments,
the mass ratio (% w/w) of detergent(s) and lipids, phospholipids,
or phosphatidylcholine is greater than 1.0, 1.2, 1.4, 1.6, 1.8,
2.0, 2.2, 2.4, 2.6, 2.8, or 3.0% w/w. For example, in some
embodiments, a composition herein can include 5% w/w sodium
deoxycholate and 4% w/w phosphatidylcholine. In some embodiments,
the ratio of % w/v of detergent(s) and lipids, phospholipids or
phosphatidylcholine is greater than 1.0, 1.2, 1.4, 1.6, 1.8, 2.0,
2.2, 2.4, 2.6, 2.8, or 3.0% w/v. Furthermore, in some embodiments,
the ratio of % v/v of detergent(s) and lipids, phospholipids or
phosphatidylcholine is greater than 1.0, 1.2, 1.4, 1.6, 1.8, 2.0,
2.2, 2.4, 2.6, 2.8, or 3.0% v/v.
[0095] In some embodiments, the compositions herein do not include
phosphatidylcholine, phospholipids, or lipids. In some embodiments,
the compositions herein do not include lipase or colipase, lipase
and colipase, a platelet activating factor inhibitor, or all
enzymes. In some embodiments, the compositions herein do not
include testosterone, prostaglandins, progesterone, estrogen, or
steroids. In some embodiments, the compositions herein do not
include tyloxapol or another alkylaryl polyether alcohol. In some
embodiments, the compositions herein do not include carprofen, a
non-steroidal anti-inflammatory compound, or any anti-inflammatory
compound. In some embodiments, the compositions herein do not
include lipids, oils, fatty acids, triglycerides or polyacrylamides
or a combination thereof. Any of the above or all of the above may
be excluded from the composition herein.
[0096] The compositions herein can be formulated for various types
of delivery e.g., topical, subcutaneous, subdermal,
intra-adipocyte, intramuscular etc. by any means known in the art.
Such formulations can be in the form of a tablet, powder, gel,
solution, cream, vapor, ointment, etc. Preferable, the
pharmacologically active detergent(s), more preferably bile acid(s)
and/or bile salt(s), more preferably deoxycholic acid or salt
thereof, or more preferably sodium deoxycholate, are formulated
into a solution. Preferably, such solution is aqueous. The term
"aqueous" as used herein refers to a solution which is a homogenous
mixture prepared by dissolving a solid or a liquid in water such
that the molecules of the solute or dissolved substance are
dispersed among those of water. Pharmacologically acceptable
aqueous vehicles for the compositions of the present invention can
include, for example, any liquid solution that is capable of
dissolving a detergent and is not toxic to the particular
individual receiving the formulation. Examples of pharmaceutically
acceptable aqueous vehicles include, without limitation, saline,
water, benzyl alcohol and acetic acid. Typically, pharmaceutically
acceptable aqueous vehicles are sterile.
[0097] In some embodiments, compositions herein are formulated for
veterinary applications with one or more veterinary excipients. In
some embodiments, compositions herein are formulated for cosmetic
applications with one or more cosmetic excipients. For delivery
into humans (e.g., transdermally or subcutaneously), the
compositions herein are formulated with one or more pharmaceutical
excipients.
[0098] Examples of pharmaceutical excipients include: buffers,
diluents, lubricating agents, solubilizers, solvents; surfactants,
penetration enhancers, polymers, dispersion agents, wetting agents,
emulsifying and suspending agents, and preserving agents. Examples
of dispersion agents include, but are not limited to, hyaluronidase
and collagenase. Hyaluronidase functions to augment tissue
permeability and spread or dispersion of other drugs. Collagenase
has been used to isolate adipocytes from subcutaneous fat and does
not have lytic effects on adipocytes themselves. Additionally
hyaluronidase and collagenase can facilitate healing by
accelerating reduction of necrotic tissue after treatment with the
detergent formulations of the present invention. In some
embodiments, the dispersion agents, such as collagenase, are
administered prior to the administration of the detergent(s)
herein. This may help release the adipocytes from the extracellular
matrix to enhance their exposure to the detergent(s).
[0099] Examples of wetting agents include, but not limited to,
acetylene glycols, acetylene alcohols, glycol ethers, alkylene
glycols, lower alcohols, and nonionic surface active agents.
Further, other alcohols, water-soluble organic solvents, anionic
surface active agents, cationic surface active agents, amphoteric
surface active agents, and saccharides may be used singly or in
combination of two or more thereof. Some examples of lower alcohols
include but not limited to, methanol, ethanol, 1-propanol,
2-propanol, 1-butanol, 2-butanol, isobutyl alcohol, tert-butyl
alcohol, 1-pentanol, 2-pentanol, 3-pentanol, 2-methyl-1-butanol,
isopentyl alcohol, tert-pentyl alcohol, 3-methyl-2-butanol,
neopentyl alcohol and water-soluble organic solvents such as
ethylene glycol, diethylene glycol, triethylene glycol, propylene
glycol, tetraethylene glycol, polyethylene glycol, dipropylene
glycol, polypropylene glycol, hexylene glycol, thiodiglycol,
glycerin, and 1,2,6-hexanetriol. In preferred embodiments, an
excipient is an organic solvent, more preferably organic alcohols,
or more preferably benzyl alcohol.
[0100] Examples of lubricating agents include, but not limited to,
talc, magnesium stearate, stearic acid, and silica gel.
[0101] Examples of the preserving agents include, but not limited
to, benzalkonium chloride, benzethonium chloride, chlorobutanol,
phenylethyl alcohol, paraoxybenzoic acid esters, and the like. Few
examples of penetration enhancers are, but not limited to,
dimethylsulfoxide, n-decyl methyl sulfoxide, N,N-dimethylacetamide,
N,N-methyl-2-pyrrolidone, and octylphenylpolyethylene glycols.
[0102] Examples of suitable emulsifiers include, but not limited
to, sodium lauryl sulfate, sodium cetyl stearyl sulfate, sucrose
stearate, and Polysorbate 80.
[0103] Examples of anionic surfactants include, but are not limited
to, fatty acid soaps, N-acylamino acids, alkyl ether carboxylates,
acylated peptides, alkylbenzenesulfonates,
alkylnaphthalenesulfonates, naphthalenesulfonate-formalin
polymerization condensates, melaminesulfonate-formalin
polymerization condensates, dialkylsulfosuccinic ester salts,
alkylsulfoacetates, .alpha.-olefinsulfonates, N-acylmethyltaurine,
sulfated oil, higher alcohol sulfuric ester salts, secondary higher
alcohol sulfuric ester salts, alkylether sulfates, secondary higher
alcohol ethoxysulfates, polyoxyethylene alkylphenyl ether sulfates,
monoglysulfates, fatty acid alkylolamide sulfuric ester salts,
alkyl ether phosphoric ester salts, and alkylphosphoric ester
salts.
[0104] Examples of nonionic surfactants include, but are not
limited to, polyoxyethylene alkyl ethers, single chain length
polyoxyethylene alkyl ethers, polyoxyethylene secondary alcohol
ethers, polyoxyethylene alkylphenyl ethers, polyoxyethylene sterol
ethers, polyoxyethylene lanoline derivatives, ethylene oxide
derivatives of alkylphenol-formalin condensates, polyoxyethylene
polyoxypropylene block polymers, polyoxyethylene polyoxypropylene
alkyl ethers, polyoxyethylene glycerol fatty acid esters,
polyoxyethylene sorbitan fatty acid esters, polyoxyethylene
sorbitol fatty acid esters, polyethyleneglycol fatty acid esters,
fatty acid monoglycerides, polyglycerol fatty acid esters, sorbitan
fatty acid esters, propyleneglycol fatty acid esters, sucrose fatty
acid esters, fatty acid alkanolamides, polyoxyethylene fatty acid
amides, polyoxyethylene alkylamines, alkylamine oxides,
polyoxyethylene castor oil derivatives, polyvinylpyrrolidone,
polyvinyl alcohol, carboxymethyl cellulose, lecithin, gelatin, and
hyaluronic acid. Any and all of the above may be used in
combination with one another as appropriate.
[0105] Other examples of suitable excipients include lactose,
dextrose, sucrose, sorbitol, mannitol, starches, gum acacia,
calcium phosphate, alginates, tragacanth, gelatin, calcium
silicate, microcrystalline cellulose, polyvinylpyrrolidone,
phosphatidylcholine, cellulose, sterile water, syrup, and methyl
cellulose.
[0106] In some embodiments, a composition herein comprises: (i) a
therapeutically effective amount of one or more pharmacologically
active detergent(s) (e.g., bile acid(s) and/or bile salt(s)); (ii)
one or more pharmaceutical, veterinary, or cosmetic excipient(s);
and optionally one or more lipids, wherein the ratio of lipids and
pharmacologically active detergent(s) is up to (but optionally not
including) 1% w/w, w/v or v/v and wherein the solution does not
include lipase or colipase. When the pharmacologically active
detergent is a bile acid, in some embodiments, the bile acid is
selected from the group consisting of: deoxycholic acid, cholic
acid, chenodeoxycholic acid, 7-alpha-dehydroxylate chenodeoxycholic
acid, lithocholic acid, ursodeoxycholic acid, dihydroxytaurin acid,
trihydroxytaurine acid, and glycine conjugates of any of the above.
When the pharmacologically active detergent is a bile salt, the
salt preferably includes a cation selected from the group
consisting of: sodium (Na.sup.+), potassium (K.sup.+), lithium
(Li.sup.+), magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium
(Ba.sup.2+), strontium (Sr.sup.2+), and ammonium (NH.sup.4+) in
combination with a bile acid listed above. In some embodiment the
cation is an alkali metal, such as those selected from the group
consisting of alkali metal is sodium (Na), potassium (K.sup.+), and
lithium (Li.sup.+). In some embodiments, the cation is an alkaline
earth metal, such as those selected from the group consisting of:
magnesium (Mg.sup.2+), calcium (Ca.sup.2+), barium (Ba.sup.2+), and
strontium (Sr.sup.2+). Preferably, a solution comprises a bile acid
salt wherein the bile salt is sodium deoxycholate.
[0107] The pharmacologically active detergent(s), more preferably
bile acid(s) and/or bile salt(s), more preferably deoxycholic acid
or salt thereof, or more preferably sodium deoxycholate is
administered at various concentrations such that a therapeutically
effective amount is delivered to a subject. A therapeutically
effective amount is the amount of detergent(s) effective to reduce
the size of a subcutaneous fat deposit or reduce the amount or
appearance of loose skin. In some embodiments, a composition with a
therapeutically effective amount of detergent(s) comprises between
about 0.001 to 10, 0.01 to 5, or 0.01 and 2% w/w, w/v, or v/v of
the detergent(s). In some embodiments, a therapeutically effective
amount of the detergent(s) is less than 5, 2, 1, 0.5, 0.2, 0.1,
0.05, 0.02, or 0.01 grams of detergent(s). If one or more lipids
are also included in the composition, the mass ratio of the
detergent(s) and lipid(s) is preferably greater than 1% w/w, w/v,
or v/v. For example, a solution of the present invention can
include about 5% bile salts (e.g., sodium deoxycholate) and up to
but not including 5% w/w, w/v, or v/v lipids, phospholipids,
phosphatidylcholine.
[0108] When the composition is formulated as a solution (preferably
aqueous solution), it may be in a container, such as a syringe or a
syringe loadable container. A solution in a container or a unit
dose of a solution herein is preferably up to 500, 250, 100, 25,
10, or 2.5 mL.
[0109] Aside from the detergent(s), the compositions/solutions
herein can also include additional active ingredient(s) or second
therapeutic agent(s). In some embodiments, such second therapeutic
agents are selected from the group consisting of: anti-microbial
agents, vasoconstrictors, anti-thrombotic agents, anti-coagulation
agents, suds-depressants, anti-inflammatory agents, analgesics,
dispersion agents, anti-dispersion agents, penetration enhancers,
steroids, tranquilizers, muscle relaxants, and anti-diarrhea
agents.
[0110] Additional Active Ingredients
[0111] In some embodiments of the present invention, a composition
can be co-formulated, co-administered, and/or co-marketed with one
or more additional active ingredients, such as, for example,
anti-microbial agents, vasoconstrictors, anti-thrombotic agents,
anti-coagulation agents, suds-depressants, anti-inflammatory
agents, analgesics, dispersion agents, anti-dispersion agents,
penetration enhancers, steroids, tranquilizers, muscle relaxants,
and anti-diarrhea agents.
[0112] Examples of anti-microbial agents suitable for use in the
compositions, methods, and kits herein include, but not limited to,
anti-bactericidal agents, anti-fungal agents, anti-viral agents and
the like, and are preferably efficacious against a broad spectrum
of microbes.
[0113] Examples of anti-fungal agents that can be used with the
composition, methods, and kits herein include dithiocarbamates,
phthalimides, dicarboximides, organophosphates, benzimidazoles,
carboxanilides, phenylamides, phosphites, and the like.
[0114] Examples of anti-bacterial agents include, but are not
limited to, erythromycin, clarithromycin, penicillins,
cephalosporins, aminoglycosides, sulfonamides, macrolides,
tetracyclins, lincosides, quinolones, chloramphenicol, vancomycin,
metronidazole, rifampin, isoniazid, spectinomycin, trimethoprim,
sulfamethoxazole, penems, carbapenems, monobactams mupirocin,
neomycin sulfate bacitracin, polymyxin B, 1-ofloxacin,
tetracyclines (chlortetracycline hydrochloride, oxytetracyc line
hydrochloride and tetrachcycline hydrochoride), clindamycin
phosphate, gentamicin sulfate, benzalkonium chloride, benzethonium
chloride, hexylresorcinol, methylbenzethonium chloride, phenol,
quaternary ammonium compounds, triclocarbon, triclosan, tea tree
oil, and their pharmaceutically acceptable salts, and
pharmaceutically acceptable salts and esters thereof.
[0115] Other examples of anti-bacterial agents include, but are not
limited to, Acrofloxacin, Amoxicillin plus clavulonic acid (i.e.,
Augmentin), Amikacin, Amplicillin, Apalcillin, Apramycin,
Astromicin, Arbekacin, Aspoxicillin, Azidozillin, Azithromycin,
Azlocillin, Bacitracin, Benzathine penicillin, Benzylpenicillin,
Carbencillin, Cefaclor, Cefadroxil, Cefalexin, Cefamandole,
Cefaparin, Cefatrizine, Cefazolin, Cefbuperazone, Cefcapene,
Cefdinir, Cefditoren, Cefepime, Cefetamet, Cefixime, Cefinetazole,
Cefminox, Cefoperazone, Ceforanide, Cefotaxime, Cefotetan,
Cefotiam, Cefoxitin, Cefpimizole, Cefpiramide, Cefpodoxime,
Cefprozil, Cefradine, Cefroxadine, Cefsulodin, Ceftazidime,
Ceftriaxone, Cefuroxime, Chlorampenicol, Chlortetracycline,
Ciclacillin, Cinoxacin, Ciprofloxacin, Clarithromycin, Clemizole
penicillin, Clindamycin, Cloxacillin, Daptomycin, Demeclocycline,
Desquinolone, Dibekacin, Dicloxacillin, Dirithromycin, Doxycycline,
Enoxacin, Epicillin, Erthromycin, Ethambutol, Fleroxacin, Flomoxef,
Flucloxacillin, Flumequine, Flurithromycin, Fosfomycin,
Fosmidomycin, Fusidic acid, Gatifloxacin, Gemifloxaxin, Gentamicin,
Imipenem, Imipenem plus Cilistatin combination, Isepamicin,
Isoniazid, Josamycin, Kanamycin, Kasugamycin, Kitasamycin,
Latamoxef, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin,
Loracarbaf, Lymecycline, Mecillinam, Meropenem, Methacycline,
Methicillin, Metronidazole, Mezlocillin, Midecamycin, Minocycline,
Miokamycin, Moxifloxacin, Nafcillin, Nafcillin, Nalidixic acid,
Neomycin, Netilmicin, Norfloxacin, Novobiocin, Oflaxacin,
Oleandomycin, Oxacillin, Oxolinic acid, Oxytetracycline, Paromycin,
Pazufloxacin, Pefloxacin, Penicillin G, Penicillin V,
Phenethicillin, Phenoxymethyl penicillin, Pipemidic acid,
Piperacillin, Piperacillin and Tazobactam combination, Piromidic
acid, Procaine penicillin, Propicillin, Pyrimethamine, Rifabutin,
Rifamide, Rifampicin, Rifamycin SV, Rifapentene, Rokitamycin,
Rolitetracycline, Roxithromycin, Rufloxacin, Sitafloxacin,
Sparfloxacin, Spectinomycin, Spiramycin, Sulfadiazine, Sulfadoxine,
Sulfamethoxazole, Sisomicin, Streptomycin, Sulfamethoxazole,
Sulfisoxazole, Synercid (Quinupristan-Dalfopristan combination),
Teicoplanin, Telithromycin, Temocillin, Tetracycline, Tetroxoprim,
Thiamphenicol, Ticarcillin, Tigecycline, Tobramycin, Tosufloxacin,
Trimethoprim, Trimetrexate, Trovafloxacin, Vancomycin, and
Verdamicin.
[0116] Examples of vasoconstrictor agents that can be used with the
compositions, methods, and kits of the present invention include
dihydroergotamine, ergotamine and methysergide, and
pharmaceutically-acceptable salts thereof.
[0117] Examples of anti-thrombotic agents that can be used with the
compositions, methods, and kits of the present invention include
argatroban, iloprost, lamifiban, taprostene, tirofiban, tissue
plasminogen activator (natural or recombinant), tenecteplase (TNK),
and lanoteplase (nPA), factor VIIa inhibitors, factor Xa
inhibitors, thrombin inhibitors (such as hirudin and argatroban),
PAI-1 inhibitors (i.e., inactivators of tissue plasminogen
activator inhibitors), alpha2-antiplasmin inhibitors,
streptokinase, urokinase and prourokinase, and anisoylated
plasminogen streptokinase activator complex, anti-coagulants (e.g.,
hirudin, heparin, etc.), plasminogen activators (e.g., t-PA,
urokinase, etc.), fibrinolytic enzymes (e.g., plasmin, subtilisin,
etc.), anti-platelet-aggregation agents (e.g., prostacyclin,
aspirin, etc.) and the like.
[0118] Examples of anti-coagulation agents that can be used with
the compositions, methods, and kits of the present invention
include cilostazol, clopidogrel, ticlopidine, tirofiban,
eptifibatide, abciximab, anagrelide, dipyridamole, aspirin,
dipyridamole/aspirin, dalteparin, enoxaparin, tinzaparin, heparin
(various), danaparoid, antithrombin III, lepirudin, argatroban,
bivalirudin, warfarin, anisidione, alteplase, reteplase,
tenecteplase, drotrecogin, anistreplase, streptokinase, urokinase,
and combinations thereof.
[0119] Examples of suds-depressants that can be used with the
compositions, methods and kits of the present invention include
monocarboxylic fatty acid and soluble salts thereof. The
monocarboxylic fatty acids and their salts can have hydrocarbyl
chains of about 1 to about 50 carbon atoms, about 10 to about 24
carbon atoms, or about 12 to about 18 carbon atoms. Suitable salts
include the alkali metal salts such as sodium, potassium, and
lithium salts, and ammonium and alkanolammonium salts. Additional
suds-depressants include, for example, high molecular weight
hydrocarbons such as paraffin, fatty acid esters (e.g., fatty acid
triglycerides), fatty acid esters of monovalent alcohols, aliphatic
C.sub.18-C.sub.40 ketones (e.g., stearone), etc. Other
suds-depressants include N-alkylated amino triazines such as tri-
to hexa-alkylmelamines or di- to tetra-alkyldiamine chlortriazines
formed as products of cyanuric chloride with 1-5 or 2-3 moles of a
primary or secondary amine containing 1 to 24 carbon atoms,
propylene oxide, and monostearyl phosphates such as monostearyl
alcohol phosphate ester and monostearyl di-alkali metal (e.g.,
K.sup.+, Na.sup.+, and Li.sup.+) phosphates and phosphate esters.
The hydrocarbons such as paraffin (including mixtures of true
paraffins and cyclic hydrocarbons) and haloparaffin can be utilized
in liquid form. It is also known to utilize waxy hydrocarbons,
preferably having a melting point below about 100.degree. C. The
hydrocarbons constitute a preferred category of suds-suppressor for
detergent compositions. The hydrocarbons can include aliphatic,
alicyclic, aromatic, and heterocyclic saturated or unsaturated
hydrocarbons having from about 12 to about 70 carbon atoms. Another
example of suds suppressors comprises silicone suds suppressors.
This category includes the use of polyorganosiloxane oils, such as
polydimethylsiloxane, dispersions or emulsions of
polyorganosiloxane oils or resins, and combinations of
polyorganosiloxane with silica particles wherein the
polyorganosiloxane is chemisorbed or fused onto the silica.
Examples also include, but not limited to, silicones, and
silica-silicone mixtures. Silicones can be generally represented by
alkylated polysiloxane materials while silica is normally used in
finely divided forms exemplified by silica aerogels and xerogels
and hydrophobic silicas of various types. Silicone suds controlling
agent, DC-544, is commercially available from Dow Corning, which is
a siloxane-glycol copolymer. Other suds suppressors include
mixtures of silicone oils and 2-alkyl-alcanols. Suitable
2-alkyl-alkanols are 2-butyl-octanol which are commercially
available.
[0120] Examples of anti-dispersion agents that can be used with the
compositions, methods, and kits herein include, but are not limited
to, sucrose, glyercerol, and glycerin.
[0121] Examples of steroids that can be used with the compositions,
methods, and kits herein include, but are not limited to,
betamethasone, chloroprednisone, clocortolone, cortisone, desonide,
dexamethasone, desoximetasone, difluprednate, estradiol,
fludrocortisone, flumethasone, flunisolide, fluocortolone,
fluprednisolone, hydrocortisone, meprednisone, methylprednisolone,
paramethasone, prednisolone, prednisone, pregnan-3-alpha-ol-20-one,
testosterone, and triamcinolone, estradiol, estron, estriol,
polyestradiol, polyestriol, dienestrol, diethylstilbestrol,
dihydroergosterone, cyproterone, danazol, testosterone,
progesterone, norethindrone, levonorgestrol, ethynodiol,
norgestimate, gestanin, 3-keton-desogestrel, demegestone,
promethoestrol, testosterone, spironolactone, and esters thereof,
budesonide, rofleponide, rofleponide palmitate, ciclesonide,
momethasone furoate, fluticasone propionate, tipredane,
fluocinolone acetonide, flunisolide, flumethasone, dexamethasone,
beclomethasone dipropionate, deflazacort, cortivazol, or cortisol
and/or hydrocortisol, prednisone, fluorometholone acetate,
dexamethasone sodium phosphate, suprofen, fluorometholone, and
medrysone, optionally in their pure isomeric forms (where such
forms exist) and pharmaceutically acceptable salts thereof.
[0122] Examples of anti-inflammatory agents that can be used with
the compositions, methods, and kits herein include both steroidal
and non-steroidal anti-inflammatory agents. Suitable steroidal
anti-inflammatory agent include, but are not limited to,
corticosteroids such as hydrocortisone, hydroxyltriamcinolone
alphamethyl dexamethasone, dexamethasone-phosphate, beclomethasone
dipropionate, clobetasol valerate, desonide, desoxymethasone,
desoxycorticosterone acetate, dexamethasone, dichlorisone,
diflorasone diacetate, diflucortolone valerate, fluadrenolone,
fluclarolone acetonide, fludrocortisone, flumethasone pivalate,
fluosinolone acetonide, fluocinonide, flucortine butylester,
fluocortolone, fluprednidene (fluprednylidene)acetate,
flurandrenolone, halcinonide, hydrocortisone acetate,
hydrocortisone butyrate, methylprednisolone, triamcinolone
acetonide, cortisone, cortodoxone, flucetonide, fludrocortisone,
difluorosone diacetate, fluradrenalone acetonide, medrysone,
amciafel, amcinafide, betamethasone and the balance of its esters,
chlorprednisone, chlorprednisone acetate, clocortelone,
clescinolone, dichlorisone, difluprednate, flucloronide,
flunisolide, fluoromethalone, fluperolone, fluprednisolone,
hydrocortisone valerate, hydrocortisone cyclopentylproprionate,
hydrocortamate, meprednisone, paramethasone, prednisolone,
prednisone, beclomethasone dipropionate, betamethasone
dipropionate, triamcinolone, and mixtures thereof.
[0123] Suitable non-steroidal anti-inflammatory agents include, but
are not limited to, oxicams, such as piroxicam, isoxicam,
tonexicam, sudoxicam, and CP-14,304; salicylates, such as salicylic
acid, aspirin, disalcid, benorylate, trilisate, safapryn, solprin,
diflunisal, and fendosal; acetic acid derivatives, such as
diclofenac, fenclofenac, indomethacin, sulindac, tolmetin,
isoxepac, furofenac, tiopinac, zidometacin, acematacin, fentiazac,
zomepiract, clidanac, oxepinac, and felbinac; the fenamates, such
as mefenamic, meclofenamic, flufenamic, niflumic, and tolfenamic
acids; propionic acid derivates, such as ibuprofen, naproxen,
benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen,
indoprofen, pirprofen, carprofen, oxaprozin, pranoprofen,
miroprofen, tioxaprofen, suprofen, alminoprofen, and tiaprofenic;
and pyrazoles, such as phenybutazone, oxyphenbutazone, feprazone,
azapropazone, and trimethazone. Mixtures of these nonsteroidal
anti-inflammatory agents can also be employed, as well as the
pharmaceutically-acceptable salts and esters thereof.
[0124] Examples of analgesics that can be used with the
compositions, methods, and kits of the present invention to reduce
discomfort due to inflammation include, but are not limited to,
lidocaine, mepivacaine, bupivacaine, procaine, chloroprocaine,
etidocaine, prilocaine dyclonine, hexylcaine, procaine, cocaine,
ketamine, morphine, pramoxine, propophol, phenol, naloxone,
meperidine, butorphanol or pentazocine, or morphine-6-glucuronide,
codeine, dihydrocodeine, diamorphine, dextropropoxyphene,
pethidine, fentanyl, alfentanil, alphaprodine, buprenorphine,
dextromoramide, diphenoxylate, dipipanone, heroin
(diacetylmorphine), hydrocodone (dihydrocodeinone), hydromorphone
(dihydromorphinone), levorphanol, meptazinol, methadone, metopon
(methyldihydromorphinone), nalbuphine, oxycodone
(dihydrohydroxycodeinone), oxymorphone (dihydrohydroxymorphinone),
phenadoxone, phenazocine, remifentanil, tramadol, tetracaine, and
mixtures thereof, as well as pharmaceutically acceptable salts and
esters thereof. In preferred embodiments, a composition includes an
analgesic selected from the group consisting of lidocaine,
hydromorphone, oxycodone, morphine and pharmaceutically-acceptable
salts thereof.
[0125] Examples of tranquilizer and sedative drugs that may be
included in the compositions, methods, and kits herein include, but
are not limited to, chlordiazepoxide, benactyzine, benzquinamide,
flurazepam, hydroxyzine, loxapine, promazine, and acceptable salts
and esters thereof.
[0126] Examples of muscle relaxants that can be included in the
compositions, methods, and kits herein include, but are not limited
to, cinnamedrine, cyclobenzaprine, flavoxate, orphenadrine,
papaverine, mebeverine, idaverine, ritodrine, dephenoxylate,
dantrolene, azumolene, and pharmaceutically-acceptable salts
thereof.
[0127] Examples of anti-diarrhea agents that can be included in the
compositions, methods and kits herein include, but are not limited
to, loperamide and pharmaceutically-acceptable salts thereof.
[0128] The examples herein and other active agents can be
co-formulated or co-administered with the one or more
pharmacologically active detergents (e.g bile acids or bile salts).
When co-formulated with a detergent (e.g., bile salt) the
additional agent can be up to 20, 19, 18, 17, 16, 15, 14, 13, 12,
11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09,
0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008,
0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008,
0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0.0002, or 0.0001% w/w, w/v
or v/v or at greater than 0.0001, 0.0002, 0.0003, 0.0004, 0.0005,
0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005,
0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06,
0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 1.0, 1.25, 1.50, 1.75,
2.0, 2.25, 2.50, 2.75, 3.0, 3.25, 3.50, 3.75, 4.0, 4.25, 4.50,
4.75, 5.0, 5.25, 5.50, 5.75, 6.0, 6.25, 6.50, 6.75, 7.0, 7.25,
7.50, 7.75, 8.0, 8.25, 8.50, 8.75, 9.0, 9.25, 9.50, 9.75, 10,
10.25, 10.50, 10.75, 11, 11.25, 11.50, 1.1.75, 12, 12.25, 12.50,
12.75, 13, 13.25, 13.50, 13.75, 14, 14.25, 14.50, 14.75, 15, 15.25,
15.50, 15.75, 16, 16.25, 16.50, 16.75, 17, 17.25, 17.50, 17.75, 18,
18.25, 18.50, 18.75, 19, 19.25, 19.50, 19.75, or 20% (w/w, w/v or
v/v) of the composition.
[0129] In some embodiments, the additional active ingredient(s) are
co-formulated with the one or more pharmacologically active
detergents (e.g., bile acids or bile salts) such that they are at a
concentration in the range of from approximately 0.001-50, 0.00140,
0.01-30, 0.02-29, 0.03-28, 0.04-27, 0.05-26, 0.06-25, 0.07-24,
0.08-23, 0.09-22, 0.1-21, 0.2-20, 0.3-19, 0.4-18, 0.5-17, 0.6-16,
0.7-15, 0.8-14, 0.9-12, or 1-10% w/w, w/v or v/v of the
composition.
[0130] In some embodiments, a composition contains up to 10, 9.5,
9.0, 8.5, 8.0, 7.5, 7.0, 6.5, 6.0, 5.5, 5.0, 4.5, 4.0, 3.5, 3.0,
2.5, 2.0, 1.5, 1.0, 0.95, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6,
0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.09, 0.08,
0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007,
0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007,
0.0006, 0.0005, 0.0004, 0.0003, 0.0002, or 0.0001 grams of
non-detergent active ingredient(s) or more than 0.0001, 0.0002,
0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001,
0.0015, 0.002, 0.0025, 0.003, 0.0035, 0.004, 0.0045, 0.005, 0.0055,
0.006, 0.0065, 0.007, 0.0075, 0.008, 0.0085, 0.009, 0.0095, 0.01,
0.015, 0.02, 0.025, 0.03, 0.035, 0.04, 0.045, 0.05, 0.055, 0.06,
0.065, 0.07, 0.075, 0.08, 0.085, 0.09, 0.095, 0.1, 0.15, 0.2, 0.25,
0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85,
0.9, 0.95, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0,
6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10 grams of the additional
active ingredient(s) or at a range between about 0.0001-10,
0.0005-9, 0.001-8, 0.005-7, 0.01-6, 0.05-5, 0.14, 0.54, or 1-3
grams of non-detergent active ingredient(s).
[0131] In some embodiments, the pharmacologically active
detergent(s) (e.g., bile acids and bile salts) herein formulated
for subcutaneous or subdermal injection directly into fat deposits
or under loose skin. Formulations for injection can be presented in
unit dosage form, for example, in ampoules, syringe loadable
containers, or in multi-dose containers, with an added
preservative. The injectable formulations can take such forms as
suspensions, solutions, or emulsions in oily or aqueous vehicles,
and can contain agents such as suspending, stabilizing
anti-dispersion and/or dispersion agents. Such formulations can
further comprise active agents such as a vasodilator to prevent the
compositions herein (e.g., deoxycholic acid or salt thereof) from
entering into the vascular system.
[0132] The compositions of the present invention can be formulated
so as to provide quick, sustained or delayed release of the active
ingredient after administration to the patient by employing
procedures known in the art. In some embodiments, a composition
herein is formulated for slow release. A slow release formulation
or a biodegradable controlled release dosage forms provide a
composition for prolonging an effect of a fat-solubilizing
effective amount of a bile acids or bile salts in vivo at a desired
site of treatment. In some embodiments, the detergents herein (e.g.
bile acids or salts thereof) are formulated with a pharmaceutically
acceptable augmenting agent which is effective to further prolong
the duration of effect of the detergent(s). The controlled release
formulation can be formed into slabs, rods, pellets,
microparticles, (e.g., microspheres, microcapsules), spheroids,
pastes solution, spray, patch, etc. Such formulations can be used
to form a suspension in isotonic saline or other physiological
buffer or a solution acceptable for subdermal injection, for a
patch, for a pump, or for a depot.
[0133] The slow release formulation can be administered by
applying, implanting, inserting or injecting a composition herein
(e.g., injectable microspheres loaded with a bile acid or bile salt
in sustained release form) into a site at or adjacent to a target
site to provide treatment. In some embodiments, the composition
herein is administered to a target site using in situ gel
implantation. In some embodiments, the pharmacologically active
detergent(s) are entrapped in a polymer carrier such as, but not
limited to, poly(DL-lactide-co-glycolide);
poly(lactide-co-glycolide); poly(DL-lactide); poly(L-lactide);
poly(glycolide); poly(.epsilon.-caprolactone);
poly(DL-lactide-co-caprolactone).
[0134] Unit Dose
[0135] The present invention also contemplates unit doses of the
compositions herein. Such unit doses are preferably in a container,
a syringe or a syringe loadable container. Such unit doses can
have, for example, a total volume of up to 500, 400, 300, 200, 100,
90, 80, 70, 60, 50, 40, 30, 20, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9,
0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06,
0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008, 0.007, 0.006, 0.005,
0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008, 0.0007, 0.0006, 0.0005,
0.0004, 0.0003, 0.0002, or 0.0001 mL. In some embodiments, a unit
dose has a total volume in the range of 0.0001-500, 0.0005-400,
0.001-300, 0.005-200, 0.01-100, 0.05-90, 0.06-80, 0.07-70, 0.08-60,
0.09-50, 0.140, 0.2-30, 0.3-29, 0.4-28, 0.5-27, 0.6-26, 0.7-25,
0.8-24, 0.9-23, 10-22, 11-21, 12-20, 13-19, 14-18, or 15-17 mL per
target site. Other embodiments contemplate a unit dose with a total
volume in the range of 0.01-30, 0.02-20, 0.03-10, 0.4-5 or 0.5-1 mL
total volume. In some embodiments, a unit dose has a total volume
greater 0.0001, 0.0002, 0.0005, 0.001, 0.002, 0.005, 0.01, 0.02,
0.05, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 50, 100 mL. Preferably, a
unit dose has a total volume of up to 1.0, 0.8, 0.6, 0.4, or
0.2.
[0136] A unit dose comprises, consists essentially, or consists of
an amount of the one or more pharmacologically active detergents
(e.g., bile acid and bile salts, or more preferably deoxycholic
acid or salts thereof) in a therapeutically effective amount. Such
amount that can be determined by a person of ordinary skill in the
art and will depend, in part, on the localization of the fat
deposit or loose skin, size of the fat deposit or loose skin and
concentration of active agent(s).
[0137] In some embodiments, a unit dose includes up to 10, 9, 8, 7,
6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1,
0.09, 0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02, 0.01, 0.009, 0.008,
0.007, 0.006, 0.005, 0.004, 0.003, 0.002, 0.001, 0.0009, 0.0008,
0.0007, 0.0006, 0.0005, 0.0004, 0.0003, 0.0002, or 0.0001 grams of
the pharmacologically active detergents. In some embodiments, a
unit dose includes a range of approximately 0.00001 to 10, 0.00005
to 1.0, 0.0001 to 0.5, 0.0005 to 0.1, 0.001 to 0.05, or 0.005 to
0.01 grams. Preferably a unit dose comprises about 0.01 grams of a
bile acid or bile salt (e.g., sodium deoxycholate).
[0138] A unit dose can further include lipids such as phospholipids
or more preferably phosphatidylcholine. Such lipids can be added in
amount and concentrations identified herein. However, in preferred
embodiments, a unit dose has up to 5% w/w, w/v or v/v lipids,
phospholipids, or phosphatidylcholine. Preferably, the ratio of %
w/w of detergent(s) and % w/w of lipids (e.g., phosphatidylcholine
in a unit dose is greater than 1, greater than 1.2, 1.4, 1.6, 1.8,
2.0, 2.2, 2.4, 2.6, 2.8, or 3.0.
[0139] Uses
[0140] The compositions herein can be used to prevent or reduce the
appearance of a skin condition, prevent or reduce the symptoms of
sleep apnea, and prevent or reduce the appearance or effects of an
adipose condition in a subject. Such subject can be an animal, more
preferably a mammal, more preferably a primate (e.g., a monkey,
chimpanzee, etc.), a domesticated animal (e.g., a dog, cat, horse,
etc.), a farm animal (e.g., goat, sheep, pig, cattle, etc.), a
laboratory animal (e.g., mouse, rat, etc.), or more preferably a
human.
[0141] In some embodiments, the compositions herein are used to
prevent or reduce the appearance of a skin condition selected from
the group consisting of loose skin, skin aging, irregularities of
the skin, and wrinkles. Such methods involve administering locally
to a to a skin region of interest a composition of the invention
comprising: (i) one or more pharmacologically active detergent(s),
more preferably one or more bile acids or bile salts, more
preferably deoxycholic acid or salt thereof, or more preferably
sodium deoxycholate; (ii) a pharmaceutical, cosmetic, or veterinary
excipient, and (iii) optionally one or more lipids. The one or more
detergents are administered in a skin-tightening effective amount.
In some embodiments, up to 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1,
0.05, 0.04, 0.03, 0.02, or 0.01 grams of detergent(s) are
administered. In some embodiments, a composition includes up to 5,
4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.04, 0.03, 0.02, or
0.01% w/w, w/v or v/v of detergent(s). When one or more lipids are
optionally included, such lipids can be phospholipids or more
preferably phosphatidylcholine. The mass or volume ratio (e.g., %
w/w, w/v or v/v) of detergent(s) to lipid(s) in the composition
delivered is preferably greater than 1. In some embodiments, a
composition comprises up to 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1,
0.05, 0.04, 0.03, 0.02, or 0.01% w/w, w/v or v/v lipids,
phospholipids or phosphatidylcholine. In any of the above
embodiments, the compositions being administered are preferably in
an aqueous solution. Such solution preferably has a total volume up
to 500 mL, 100 mL, 50 mL, 10 mL or 5 mL.
[0142] The above compositions are preferably administered locally
to a target area to create an inflammatory response causing a scar
formation. The formation of a scar results in skin tightening
especially in areas where the skin is under little or no tension
and provides little resistance to scar contraction. Such treatment
may be relevant in a number of clinical scenarios such as, for
example, commonly performed fat treatments, including, but not
limited to, large volume liposuction. The latter may be associated
with post-treatment skin laxity (loose skin in areas of fat
removal) and skin surface irregularities. Thus, in some embodiments
a composition comprising a therapeutically effective amount of
detergent(s) or bile salt(s) may be administered to a liposuction
site after completion of the liposuction procedure.
[0143] In some embodiments, the target area is an area under eye,
under chin, under arm, buttock, calf, back, thigh, stomach, cheek,
brow, or any other skin regions showing aging, wrinkles, loose skin
or skin irregularity.
[0144] In some embodiment, the compositions herein are delivered to
the target area via a subdermal injection, a pump, a patch, or a
subdermal depot. In some embodiments, the compositions herein are
administered topically.
[0145] In one aspect, the compositions herein are used for reducing
a subcutaneous fat deposit in a subject. The subcutaneous fat
deposit can be associated with an adipose conditions such as, for
example, obesity, fat redistribution syndrome, eyelid fat
herniation, lipomas, lipodystrophy (including buffalo hump
lipodystrophy), dorsocervical fat, visceral adiposity, breast
enlargement, hyperadiposity, diffused body fat around trunk and
arms, fat deposits associated with cellulite, Dercum's disease,
Madelung's neck, lipedema, piezogenic nodules, launois cleret
syndrome and xanthelasma. The subcutaneous fat deposit can be
located in an area of the body, including, but not limited to,
under eye, under chin, under arm, buttock, calf, back, thigh,
stomach, cheek, brow, "love-handles", ankles, fingers, lips,
trachea, etc.
[0146] The subcutaneous fat deposits can be reduced by
administering locally to the fat deposits a composition comprising:
(i) one or more pharmacologically active detergents, more
preferably one or more bile acids or salts thereof, more preferably
deoxycholic acid or salts thereof, or more preferably sodium
deoxycholate; (ii) a pharmaceutical or veterinary excipient; and
(iii) optionally a lipid, wherein the ratio of the lipid and bile
acid or bile salt is up to 1% w/w. Preferably, the composition does
not include lipase or colipase. The above method preferably does
not include performing surgery (e.g., liposuction) on the
subject.
[0147] Preferably, the one or more pharmacologically active
detergents are administered in a fat-dissolving effective amount
such as up to 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.04,
0.03, 0.02, or 0.01 grams of detergent(s). In some embodiments, a
composition includes up to 5, 4, 3, 2, 1, 0.5, 0.4, 0.3, 0.2, 0.1,
0.05, 0.04, 0.03, 0.02, or 0.01% w/w, w/v or v/v of detergent(s).
Preferably a composition comprises between about 0.001 to 10% w/w
of detergent(s) or bile salt(s), or more preferably between about
0.01 and 5% w/w detergent(s) or bile salt(s).
[0148] When one or more lipids are optionally included, such lipids
can be phospholipids or more preferably phosphatidylcholine. The
mass or volume ratio (e.g., % w/w, w/v or v/v) of detergent(s) to
lipid(s) in the composition delivered is preferably greater than 1.
In some embodiments, a composition comprises up to 5, 4, 3, 2, 1,
0.5, 0.4, 0.3, 0.2, 0.1, 0.05, 0.04, 0.03, 0.02, or 0.01% w/w, w/v
or v/v lipids, phospholipids or phosphatidylcholine. In any of the
above embodiments, the composition being administered is preferably
in a solution or more preferably an aqueous solution. Such solution
preferably has a total volume up to 500 mL, 100 mL, 50 mL, 10 mL or
5 mL.
[0149] The subcutaneous fat deposit can be associated with an
adipose condition selected from the group consisting of obesity,
fat redistribution syndrome, eyelid fat herniation, lipomas,
Dercum's disease, lipodystrophy, buffalo hump lipodystrophy,
dorsocervical fat, visceral adiposity, breast enlargement,
hyperadiposity, diffused body fat around trunk and arms, and fat
deposits associated with cellulite. Thus, the compositions herein
can be used to treat or ameliorate conditions associated with such
conditions. In some embodiments, the composition herein are
administered locally under eye, under chin, under arm, buttock,
calf, back, thigh, or stomach of said subject to reduce a
subcutaneous fat deposit in such site.
[0150] For example, in some embodiments, a composition herein
(e.g., a pharmaceutically active detergent, or more preferably a
bile salt, or more preferably sodium deoxycholate) is use to treat
or ameliorate lipodystrophy conditions in a subject, such as a
human HIV patient. Lipodystrophy is a condition that is often a
side effect from HIV treatments (e.g., treatment with protease
inhibitors). Lipodystrophy is characterized by regional or
generalized loss of subcutaneous fat or abnormal fat redistribution
and metabolic disturbances. Signs of lipodystrophy can include a
swollen belly along with loss of tissue from the face, arms and
legs, which can give an appearance of sunken eyes and sticking-out
cheekbones. Other signs of lipodystrophy include fat accumulates on
the back of the neck, which is sometimes referred to as buffalo
humps (diffused fat accumulation). Female HIV patients may also be
treated for breast enlargement which is attributed to the
lipodystrophy syndrome.
[0151] In some embodiments, the compositions herein are used to
treat lipomas. Lipomas are localized fat accumulations that are
benign neoplastic growth. There are various forms of lipomas and,
in some embodiments; the compositions herein are used to treat
multiple familial lipomatosis.
[0152] In some embodiments, the compositions herein are used to
prevent, treat or ameliorate an adipose in an animal such as a cat,
a dog or a horse.
[0153] In some embodiments, the compositions herein are used to
treat obstructive sleep apnea. Obstructive sleep apnea is
characterized by repetitive pauses in breathing during sleep due to
the obstruction and/or collapse of an upper airway (throat),
usually accompanied by a reduction in blood oxygen saturation, and
followed by an awakening to breathe. It is a dangerous (sometimes
life threatening) condition that often affects obese people. Obese
people have a large amount of fat around their trachea, and this
fat may cause their airway to collapse when their muscles relax
during sleep. In one embodiment, the compositions herein (e.g.,
sodium deoxycholate) are used to treat obstructive sleep apnea by
dissolving fat around the trachea. In such embodiments, a
composition of the invention is administered locally (e.g., via
injection) to a target site of fat around the trachea in a
therapeutically effective amount.
[0154] For the treatment of an adipose condition, the compositions
herein (e.g., pharmacologically active detergent(s), bile salt(s),
or sodium deoxycholate) are preferably administered locally to the
site of fat accumulation. Localized delivery can be made via, e.g.,
subcutaneous or transdermal injection, external or internal pump,
dermal patch, subcutaneous depot, or any other means known in the
art.
[0155] In some embodiments, a composition herein is delivered via a
dermal patch. A dermal patch is a self-adhesive unit, and is worn
on a patient's body. It delivers small doses of a drug into the
skin, where it then diffuses into the skin. The patch incorporates
a series of thin, flexible films: a backing layer, a drug
reservoir, a rate-controlling film and an adhesive. Enhancers may
be added to further increase drug permeation through the skin. The
patch can be coupled with a low-level electrical energy to actively
transport drugs through intact skin.
[0156] In some embodiments, a composition herein is delivered via
an external or internal pump. A pump is a specialized device, which
delivers drug into the body via a small catheter. For example, an
infusion pump can be programmed to deliver drugs at precise dosages
and delivery rates. These pumps may have a feedback device that
controls drug delivery according to need. A size of the pump
depends on an amount of a drug and intended length of
treatment.
[0157] In some embodiments, a composition herein is delivered via a
depot. A depot is a non-biodegradable and an osmotically driven
implant which is used to enable delivery of drugs for therapy.
Powered by osmosis, the depot incorporates a miniature metal alloy
cylinder and can provide continuous drug delivery from days to up
to one year. In some embodiments, the compositions herein may be
administered parenterally. Parenteral routes of administration
involve injections into various compartments of the body such as
but not limited to, intravenous, subcutaneous, intramuscular,
intraperitoneal and the like.
[0158] In one example, two injections of up to 1 mL each are
administered under the chin wherein each injection comprises
between 0.005 and 0.5, or more preferably between 0.002 and 0.08
grams sodium deoxycholate. In another embodiment 3 treatments of 2
mL 1% w/w sodium deoxycholate are administered to a cheek area.
[0159] In any of the embodiments herein, a therapeutic regimen can
include administering one or more unit doses to a target site. A
target site can be for example 0.1 cm.sup.2, to about 5 cm.sup.2.
The compositions herein may be administered at the same, adjacent,
or nearby target sites at various intervals, dosages, volumes, as
disclosed herein. When delivered, the compositions can be
administered at various levels below the dermis, including, for
example, 0.1-4, 0.2-3.5, 0.3-3, 0.4-2.5, or 0.5-2 inches below the
dermis.
[0160] For example, in some embodiments up to 100, 10, 9, 8, 7, 6,
5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09,
0.08, 0.07, 0.06, 0.05, 0.04, 0.03, 0.02 or 0.01 mL of a solution
is delivered locally to a target site (e.g., a site of fat
accumulation or loose skin) at a time. The solution can comprise,
consist essentially of, or consist of one or more pharmacologically
active detergent(s), (e.g. bile acids or salts thereof, or more
preferably deoxycholic acid or salts thereof, more preferably
sodium deoxycholate) wherein the solution contains up to 5, 4, 3,
2, 1% w/w of the detergent(s). Such a solution can include up to 5%
w/w, w/v, or v/v lipids, or phospholipids, or phosphatidylcholine,
or in some embodiments no phosphatidylcholine. In some embodiments,
the compositions (e.g., solutions) herein include no lipase or no
colipase, or no colipase and lipase. In some embodiments, the
compositions (e.g., solutions) herein include no enzymes. In some
embodiments up to 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7,
0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.09, 0.08, 0.07, 0.06, 0.05, 0.04,
0.03, 0.02 or 0.01 grams of the detergent(s) herein (preferably
bile acids or bile salts) are administered locally to a target site
at a time. In other embodiments more than 0.001, 0.002, 0.003,
0.004, 0.005, 0.006, 0.007, 0.008, 0.009 or 0.01 g is delivered to
a target site at a time. Generally, the total volume, unit dose and
number of treatments administered will vary depending on the amount
of fat in a target site, the location of the target site, type of
fat composition, and desired results. In general, the greater is
the amount of fat being treated, the greater is the dose that is
administered. Also, the greater the amount of loose skin in a
target area, the greater the dose or the more number of injections
that will be delivered. It should be noted that while the
compositions and unit dosages herein may be administered into an
individual as part of a treatment regimen, they are not necessarily
actively removed from the individual as part of the treatment
regimen (e.g., via suction).
[0161] In any of the embodiments herein, a therapeutic regimen can
include administration to a target site at least 1, 2, 3, 4, 5, 6,
7, 8, 9, or 10 unit doses. Such unit doses can be delivered
simultaneously or over a period of time. For example administration
can occur once, twice, three or four times an hour, a day, a week,
a month, or a year. In some embodiments, multiple administrations
involve between 1-100, 2-50, 3-30, 4-20, or 5-10 administrations to
a target site an hour, a day, a week, a month or a year. In some
embodiments, multiple administrations involve up to 10, 9, 8, 7, 6,
5, 4, 3, or 2 administrations to a target site a year, a week, a
day or an hour. The total number of administrations in a
therapeutic regimen can be completed with a period of 1 year, 6
months, 5 months, 4 months, 3 months, 2 months, 1 month, 3 weeks,
2, weeks, or 1 week or less.
[0162] In any of the embodiments herein, the methods do not include
the surgical removal of one or more localized fat deposits in a
subject. A non-surgical method of fat reduction does not include
liposuction, lipoplasty or suction lipectomy. For example, in some
embodiments, the non-surgical methods herein do not include
liposuction. In some embodiments, the methods herein also exclude
non-invasive means for reducing fat, e.g., ultrasonification. In
other embodiments, non-invasive means can be used in conjunction
with the compositions herein.
[0163] Any of the treatments disclosed may be supplemented by
further administering to the patient additional active agent(s).
Such additional agent(s) can be administered separately or in
combination with the compositions herein. A second agent can be
administered locally or systemically. In some embodiments, a second
agent is co-formulated with the detergent(s). In other embodiments,
second agents are administered prior to, simultaneous with, or
after the administration of the detergents.
[0164] Kits
[0165] FIG. 11 is an illustration of a kit 101 for non-surgical
reduction of subcutaneous fat and/or tightening of loose skin. Kit
101 includes one or more containers 102. For example, a first
container 102 can include a therapeutically effective amount of a
pharmacologically active detergent (e.g., bile acids and salts
thereof) and up to 5% w/v phosphatidylcholine and a second
container 102 can include a therapeutically effective amount of a
pharmacologically active detergent and no phosphatidylcholine.
Containers 102 are preferably syringe loadable. Containers 102 can
each hold one or more unit doses. For example, a container 102 can
hold up to 500, 100, 20, 10, 5 or 1 mL.
[0166] In some embodiments, a container 102 can include one or more
additional active ingredients, either independently of or in
combination with the one or more detergents herein. Examples of
additional active ingredients include antimicrobial agents,
anti-thrombotic agents, anti-coagulation agents, suds-depressants,
anti-inflammatory agents, analgesics, anesthetics, anti-dispersion
agents, dispersion agents, penetration enhancers, steroids,
tranquilizers, muscle relaxants, and anti-diarrhea agents.
[0167] In one example, a container 102 includes up to 5% w/v of
phospholipids (e.g. phosphatidylcholine) or no phospholipids (e.g.
phosphatidylcholine) and up to 10, 5.0, 1.0, or 0.5% w/w
detergent(s).
[0168] In one example, a container 102 contains more than 0.01,
0.1, 1.0, 2.0, 3.0, 4.0, or 5.0% w/w, w/v or v/v pharmacologically
active detergent(s) (e.g., bile acids or bile salts) and up to 5%
w/v phospholipids (e.g. phosphatidylcholine) or no phospholipids
(e.g. phosphatidylcholine).
[0169] The solution of container 102 is administered according to
the instructions for use 103. Instructions for use 103 can provide
dosing instructions which may depend upon, for example, location of
target site, animal being treated, desired results, size of target
site, concentration of detergent(s) in composition, etc. Preferably
instructions for use 103 are for the treatment of an animal such as
a human, a dog, a cat, or a horse. Instructions for use 103 can
also include information for treatment of other domesticated
animals and/or farm animals. Instruction for use 103 may also
include information on the use of the compositions herein to treat
specific target sites, such as, e.g., fat deposits or areas of
loose skin localized under eye, under chin, under arm, buttock,
cheek, brow, calf, back, thigh, ankle, or stomach. In some
embodiments, instruction for use 103 detail an explanation for use
of the compositions herein to treat a fat condition of obesity, fat
redistribution syndrome, dorsocervical fat, visceral adiposity,
breast enlargement, hyperadiposity, eyelid fat herniation, lipomas,
lipodystrophy, buffalo hump lipodystrophy, diffused body fat around
trunk and arms, or fat deposits associated with cellulite.
[0170] In some embodiments, instructions for use 103 detail an
explanation for use of the compositions herein to treat a skin
condition selected from the group consisting of loose skin, skin
aging, irregularities of the skin, and wrinkles.
[0171] Instruction for use 103 may include information regarding
proper diluents and volumes for dilution, if any, of the container
102. The instructions for use 103 may also provide information
regarding the proper administration of the compositions herein,
such as frequency and dosage of administration.
[0172] Kit 101 can comprise alternatively or in addition to
container 102 one or more syringes 104 or other suitable delivery
devices (e.g., patches, subcutaneous depots) for delivering the
compositions herein (e.g. those in container 102) to a target site
of fat accumulation or loose skin. In some embodiments, syringe or
other delivery device 104 may be preloaded with one or more unit
doses of the compositions herein.
[0173] Preferably, a kit includes one or more syringes for local
subcutaneous injection of a solution having total volume up to 100
mL containing between 0.1-10% of a bile acid or a salt, or more
preferably deoxycholic acid or salt thereof or sodium deoxycholate.
The solution preferably does not contain lipase or colipase or
both. The solution preferably contains up to or no
phosphatidylcholine.
[0174] The invention contemplates kits having a first container
comprising a pharmacologically active detergent and up to 5% w/v
phosphatidylcholine, as well as written instructions for reducing
subcutaneous fat deposits in a mammal without the use of surgery.
Preferably, the kits herein may be used to reduce fat deposits in a
variety of mammals such as, for example, a human, a horse, a dog,
or a cat. In some embodiments the mammal is a human.
[0175] In some preferred embodiments, the first container has a
total volume of up to 500 mL and/or is provided as an injectable
formulation. In other preferred embodiments, the first container
may contain a % w/v of detergent greater than the % w/v of
phosphatidylcholine or may contain no phosphatidylcholine. In one
preferred embodiment, the present invention provides the detergent
at a concentration above its critical micellar concentration (CMC).
The kits may comprise a variety of pharmacologically active
detergents such as, for example, a lipophilic detergent, a
hydrophilic detergent, an ionic detergent, a non-ionic detergent, a
glyceride, a bile salt, and a zwitterionic detergent. In a more
preferred embodiment, the active detergent is a bile salt, most
preferably sodium deoxycholate. A first container in the kit herein
may, in some embodiments include up to 3 g detergent. In other
embodiments, a first container in the kit herein may include more
than 0.0002 g detergent. In any of the embodiments herein the first
container may further include a second detergent.
[0176] Preferably, the first container may further comprise a
second therapeutic agent such as, for example, an anti-microbial
agent, a vasoconstrictor, an anti-thrombotic agent, an
anti-coagulation agent, a suds-depressant, an anti-inflammatory
agent, an analgesic, a dispersion agent, an anti-dispersion agent,
a penetration enhancer, a steroid, a tranquilizer, a muscle
relaxant, and an anti-diarrhea agent. In some embodiments the
second therapeutic agent is an analgesic, anti-microbial agent, or
an anti-inflammatory agent. More preferably, the second therapeutic
agent is an analgesic, or most preferably lidocaine. In another
embodiment, the kit provides a second container comprising the
second therapeutic agent as described herein.
[0177] One embodiment of the present invention contemplates a kit
herein for reducing fat deposits under the eye, chin, or arm, as
well as the buttock, calf, back, thigh, ankle, or stomach of a
mammal. In another embodiment, the kit may reduce specific types of
fat deposits such as, for example, eyelid fat herniation, lipomas,
lipodystrophy, buffalo hump lipodystrophy, diffused body fat around
trunk and arms, or fat deposits associated with cellulite.
[0178] Business Methods
[0179] The methods and the kits disclosed herein can be used to
perform business services and/or sell business products.
[0180] In some embodiments, the present invention contemplates a
business method that sells the kits herein or provides treatment
services. For example, the business can make a formulation based on
the compositions described herein. The business methods herein can
then manufacture kits containing such formulations and sell such
kits in exchange for a fee. In some embodiments, the business
method licenses a third party to manufacture the kit. In some
embodiments, the business contracts a sales support to sell such
kits.
[0181] The business can alternatively or in addition perform
treatment services in exchange for service fees. The service can be
provided directly to patients and the fee can vary depending on the
length of the procedure and/or amount of active detergents
used.
[0182] It is understood that there are numerous other embodiments
and methods of using the present invention that will be apparent
embodiments to those of ordinary skill in the art after having read
and understood this specification and examples. The following
examples are meant to illustrate one or more embodiments of the
invention and are not meant to limit the invention to that which is
described below.
EXAMPLES
Example 1
Sodium Deoxycholate and Phosphatidylcholine Formulations
[0183] Phosphatidylcholine bile salt formulation (PBF) (5.0% highly
purified soy derived PC, 4.75% sodium deoxycholate, and 0.9% benzyl
alcohol, in sterile water, Table 2) was obtained from Hopewell
Pharmacy, Hopewell, N.J. Sodium deoxycholate and Triton X-100
detergent (Triton, alkylaryl polyether alcohol) were obtained from
Sigma-Aldrich Corp. (St. Louis, Mo.). Empigen BB detergent
(Empigen, lauryldimethylbetaine) was obtained from Calbiochem,
Biosciences, Inc., (La Jolla, Calif.). An injectible PBF solution
was made according to Table 2 below. Stock reagents (5% dilutions)
were prepared in PBS buffer. TABLE-US-00002 TABLE 2 Injectable PBF
Phosphatidylcholine 5.00% (w/v) Sodium deoxycholate 4.75% Benzyl
alcohol 0.90% Water 100 mL
[0184] The molecular structure of (a) phosphatidylcholine, (b)
sodium deoxycholate and (c) benzyl alcohol are depicted in FIG.
1.
Example 2
Effects of Sodium Deoxycholate and Phosphatidylcholine Solutions in
Cultured Cells
[0185] To measure cell viability after detergent treatment, HaCaT
human keratinocyte cells were cultured in DMEM (Dulbecco's modified
Eagle's medium) supplemented with 10% fetal calf serum, penicillin,
and streptomycin. HaCaT cells were cultured in 6 well plates and
incubated with 0, 0.005, 0.050 or 0.500% of the PBF or sodium
deoxycholate for 30 min at 37.degree. C. prior to determination of
cell viability using the MTS assay, which uses a tetrazolium
compound that produces a color change when bioreduced by
metabolically active cells (CellTiter 96 Aqueous Non-Radioactive
Cell Proliferation Assay, Promega, Corp. Madison, Wis.). Cell
viability was determined by an absorbance spectrophotometer (at 490
nm) after 4 hour incubation with the assay at 37.degree. C. To
determine cell viability in fresh tissue, fat specimens were
incubated for 4 hours in 24 well plates with stock reagents and the
MTS assay. Tissue specimens were then visualized for color change
and the amount of MTS in their supernatants was measured by
absorbance (at 490 nm). All studies were performed in triplicate.
Absorbance at 490 nm (OD 490) is proportional to the number of
living cells in the culture. There was comparable OD 490 in the
control and 0.005% dilutions of both compounds (FIG. 2a),
indicating little effect of these substances on cell viability at
this concentration. Cell viability progressively decreased at 0.05%
and 0.5% concentrations of both solutions.
[0186] Cell lysis in response to detergent treatment was determined
in HaCaT cells incubated with the reagents at the indicated cell
dilutions for 30 min at 37.degree. C. Lysis in cultured cells was
measured using a lactate dehydrogenase (LDH) assay and within
tissue using calcein, a fluorescent marker retained in cells with
intact cell membranes. The LDH assay measures the activity of LDH,
which is a cytosolic enzyme released when cells are lysed. Lactate
dehydrogenase release was measured by absorbance (at 490 nm) after
a 1 hour incubation with the LDH assay as recommended by the
manufacturer (CytoTox 96 Non-Radioactive Cytotoxicity Assay,
Promega). All studies were performed in triplicate. LDH release is
directly proportional to absorbance at 490 nm (OD 490). Because
penetration into intact tissues may be likely a limiting factor,
cell cultures were used to determine the dilutions of the reagents
(PBF and deoxycholate) necessary to affect cells. As is illustrated
in FIG. 2a, sodium deoxycholate profoundly decreased the viability
of cultured cells approximately equal to the complete PBF. This
finding was reproduced in tissue by exposing porcine fat to PBF and
deoxycholate (FIG. 3). These results support the unexpected
observation that sodium deoxycholate plays a major, active role in
the PBF. There was minimal LDH release from control cells and those
incubated with 0.005% dilutions of both compounds (FIG. 2b).
Moreover, both the PBF and deoxycholate treated cell cultures
demonstrated a concentration dependent increase in cell lysis (FIG.
2b). Moreover, the direct lytic effects observed in cultured cells
treated with these agents suggest activity independent of
endogenous lipase. There was progressively more LDH released at
0.05% and 0.5% of the PBF and deoxycholate. Comparing the effects
of the PBF to deoxycholate in cell culture led to the surprising
result that deoxycholate caused similar loss of cell viability, but
less cell lysis. These data unexpectedly demonstrate that
deoxycholate acts as the active component in PBF.
Example 3
Effects of Sodium Deoxycholate and Phosphatidylcholine Solutions in
Porcine Tissue
[0187] Porcine tissue was obtained immediately after sacrifice,
shaved, and placed on ice for a maximum of four hours before use.
Fat specimens were obtained by removing the epidermis and dermis of
a punch biopsy with a scalpel and trimmed. Fat specimens were
loaded with calcein dye by incubating 1 hour at 37.degree. C. with
Calcein-AM (Sigma). Stock reagents were added to the fat specimens
and incubated for 30 min at 37.degree. C. with gentle agitation.
Calcein retention was determined by tissue fluorescence using
purple (411 nm) light and visually observing the emitted green (500
nm) light using an emission filters.
[0188] Histology was performed by injecting stock reagent solutions
(0.5 mL) into full thickness porcine skin at various levels
(epidermis, dermis, and subcutaneous tissue) with 1.0 mL syringes
and 30-gauge, 0.5 inch needles. Needle depth was visualized along
the margin of the porcine tissue with the intent of saturating the
target tissue. One hour after incubation with PBS at 37.degree. C.,
multiple 5.0 mm biopsy specimens were obtained from the injected
sites, each condition performed in triplicate. Tissue was fixed in
formaldehyde, paraffin-embedded, and stained with
hematoxylin-eosin. Specimens were evaluated by a board-certified
dermatopathologist who was blinded to the treatment protocol.
[0189] Fresh porcine skin was used to determine if the effects of
these detergent substances on cultured cells were similar in
tissue. FIG. 3a demonstrates the production of dark purple pigment
(indicating viable cells) in fat tissue treated with the PBS buffer
(negative control) using the MTS assay. The PBF and 5% solutions of
deoxycholate and Triton detergent (positive control) demonstrated a
comparable loss of purple dye (indicating cell death) in the
treated fat specimens. The difference in fat cell viability between
the solutions was quantified by measuring the absorbance (at 490)
of the supernatants collected from the treated fat specimens (FIG.
3b). All reagents had significant effects on the fat cell viability
of fresh tissue.
[0190] Cell lysis was confirmed using a calcein dye release assay.
Calcein becomes fluorescent after hydrolysis and is retained in
cells that have intact cell membranes. Because it does not label
dead cells and is lost under conditions that cause cell lysis, loss
of green fluorescence in fat tissue samples loaded with the dye
calcein indicates cell lysis (FIG. 4). Samples treated with the
deoxycholate, PBF, and Triton detergent (positive control)
exhibited similar loss of fluorescence.
[0191] The histologic changes resulting from injection of PBF,
deoxycholate, and Empigen, are shown in FIG. 5. Phosphatidylcholine
bile salt formulation (FIG. 5b) and deoxycholate (FIG. 5d) produced
histologic effects similar to those caused by Empigen (FIG. 5g) and
Triton (not shown), two well-characterized laboratory detergents.
These changes were apparent in both fat and muscle. Marked blurring
and dissolution of adipocyte cell membranes with disruption of its
normal lobular architecture were seen, after injection of both the
PBF (FIG. 5b) and deoxycholate (FIG. 5d). FIG. 5f demonstrates
muscle fiber disarray and atrophy after PBF injection. Similar
changes in muscle tissue were visible in the specimens treated with
deoxycholate and the Triton and Empigen detergents. There were no
changes in the epidermis, dermis, or adnexal structures after
injection of the reagents with the exception of Empigen, which
caused loss of fibroblast nuclear staining and hyalinization of
dermal collagen. Moreover, from clinical reports, it is apparent
that a brisk inflammatory response, such as erythema and edema,
occurs after injection with the detergents. Repeated inflammation
can potentially lead to fibrosis, especially after multiple
injections. Fibrosis has been reported in several patients who
developed firm nodules at injection sites after PBF administration
that eventually resolve over several months.
[0192] Histologic findings reveal that the injectable PBF and
deoxycholate alone cause architectural disruption in fat and
muscle, but had no apparent affect on the epidermis, dermis, or
adnexae (FIG. 5). However, Empigen BB, a potent laboratory
detergent, had profound histologic effects on dermal collagen
(connective tissue). Alternatively, fat and muscle can be more
sensitive to detergent treatment than these other structures at the
tested concentrations (similar to those used in clinical
practice).
[0193] Through a series of laboratory experiments utilizing fresh
tissue specimens and cell cultures, it has been demonstrated that
the PBF popularly used in subcutaneous injections for fat
dissolution works primarily by causing non-specific lysis of cell
membranes. Cell membranes are constituents of all tissue types;
specifically, the present inventor demonstrated that these
detergents cause solubilization of fat, muscle and connective
tissue. Therefore sodium deoxycholate, the bile salt component of
the formula used to dissolve the phosphatidylcholine, was the major
active ingredient of these prior art formulations. This conclusion
is supported by the fact that pharmacologically active detergents,
such as bile salts are potent solubilizers of cell membranes.
Example 4
Clinical Experience with Sodium Deoxycholate Compositions
[0194] Patients having lipomas, benign, isolated collections of
adipose tissue, were injected with sodium deoxycholate (DC)
solutions without phosphatidylcholine directly into the lipoma. The
results of this study demonstrate that the detergent effects of
deoxycholate seen on fat in animal tissues are reproducible
clinically in humans. All injected lipomas were reduced in size
after at least one treatment with varied concentrations of
deoxycholate (Table 3). A lipoma from one patient, injected with 1%
DC, was excised after treatment and pathological and histological
analysis performed. Within the excised lipoma, necrosis is visible
grossly (FIG. 6a) with a well demarcated area of hemorrhage and
necrosis on the lateral edge extending into the middle of the
lipoma fat which contrasts with the normal lipoma fat which is
lighter in color. Histological analysis (FIG. 6b) reveals a well
defined area of hemorrhage and necrotic fat as well as a
significant inflammatory reaction which contrasts to the adjacent
normal round clear fat cells. TABLE-US-00003 TABLE 3 Reduction in
size of lipomas after DC treatment Size (cm) Size(cm) Total
Treatments Lipoma Pre-treatment Post-treatment (% DC injected) 1
2.00 .times. 1.00 1.25 .times. 0.50 2 (2.5%) 2 2.00 1.50 .times.
0.50 3 (5% and 2.5%) 3 2.00 .times. 2.50 2.00 .times. 1.00 3 (5%
and 2.5%) 4 4.00 .times. 1.75 2.50 .times. 2.00 2 (1%) 5 2.00
.times. 1.75 1.25 2 (1%) 6 2.80 0.50 1 (5%) 7 1.00 Imperceptible 1
(1%)
Example 5
Treatment of Lipodystrohy in a HIV Patient with Sodium Deoxycholate
Compositions
[0195] An HIV lipodystrophy patient with symmetrical fat deposits
in his face just vental of his master muscle (sort of in his
cheeks, like acorns in the mouth of a chipmunk) was injected with
1% deoxycholate. At 1 month follow up, there was a 50% apparent
reduction in deposit volume on both sides. The results of this
study demonstrate that the effects of deoxycholate seen on fat in
animal tissues are reproducible clinically in humans.
Example 6
Effects of Sodium Deoxycholate and Sodium
Deoxycholate-Phosphatidylcholine Solutions on Melanocytes
[0196] Melanocytes in melanocyte media on a 96-well dish were
differentiated at 37.degree. C. in a humidified, 5% CO.sub.2
incubator for a week. Media was aspirated and washed with
1.times.PBS once. Cells were treated with different concentrations
of DC (from 0 to 0.5%) with or without 1% PC in, 1.times.PBS. Each
treatment was done in triplicates (Table 4). The plates were
incubated at 37.degree. C. in a humidified, 5% CO.sub.2 incubator
for 30 min. The incubated plates were aspirated and washed with
1.times.PBS.
[0197] 10 mL of MTS
(3-(4,5-dimethyl-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-te-
trazolium, inner salt) assay solution was prepared (mixed 40 uL MTS
(Promega, Madison Wis.) per Mel 1.times.PBS). 100 ul of MTS
solution was added to each well of the plates. The plates were
incubated at 37.degree. C. incubator (non-humified) for 120 min. OD
was taken with spectrophotometer at 490 nm.
[0198] FIG. 7 illustrates the survival of melanocytes in sodium
deoxycholate solution and sodium deoxycholate with 1%
phosphatidylcholine solution. A375P is a melanoma cell line (P
represents poorly metastatic). On y-axis, 1 represents 100%
survival when no DC is added. All % concentrations are w/v %. LD50
(concentration at which 50% of the fat cells are dead) of only
sodium deoxycholate solution for melanocyte cell line was found to
be around 0.04% sodium deoxycholate. The addition of sodium
deoxycholate with 1% phosphatidylcholine solution increases the
LD50 by .about.5-6 fold.
[0199] Results of this study demonstrate that addition of 1%
phosphatidylcholine inhibits apoptosis in vitro i.e., the presence
of PC makes it 5-times harder for DC to kill the melanocytes,
showing that PC detracts from, and thus does not enhance the cell
killing power of DC. TABLE-US-00004 TABLE 4 Melanocytes after
treatment with DC only and DC + 1% PC Melanocytes Melanocytes
surviving in Percentage surviving in Percentage DC DC only Average
surviving % DC + 1% PC Average surviving % 0% 0.163 0.166667 1
0.283 0.342333 1 0.16 0.326 0.177 0.418 0.001 0.158 0.155333 0.932
0.248 0.356333 1.040896 0.148 0.239 0.16 0.582 0.005 0.128 0.113333
0.68 0.226 0.263667 0.770204 0.109 0.259 0.103 0.306 0.01 0.124
0.112333 0.674 0.197 0.220667 0.644596 0.107 0.223 0.106 0.242 0.05
0.056 0.047333 0.284 0.207 0.232 0.677702 0.038 0.214 0.048 0.275
0.1 0.009 0.013 0.078 0.191 0.182333 0.532619 0.014 0.168 0.016
0.188 0.5 0.001 0.004 0.024 0.012 0.006333 0.0185 0.009 0.003 0.002
0.004
Example 7
Effects of Sodium Deoxycholate and Sodium
Deoxycholate-Phosphatidylcholine Solutions on Adipocytes
[0200] Adipocytes in adipocyte media on a 96-well dish were
differentiated at 37.degree. C. in a humidified, 5% CO.sub.2
incubator for a week. Media was aspirated and washed with
1.times.PBS once. Cells were treated with different concentrations
of DC (from 0 to 0.5%) with or without 1% PC in 1.times.PBS. Each
treatment was done in triplicates (Table 5). The plates were
incubated at 37.degree. C. in a humidified, 5% CO.sub.2 incubator
for 30 min. The incubated plates were aspirated and washed with
1.times.PBS.
[0201] 10 mL of MTS
(3-(4,5-dimethyl-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-te-
trazolium, inner salt) assay solution was prepared (mixed 40 uL MTS
(Promega, Madison Wis.) per 1 mL 1.times.PBS). 100 ul of MTS
solution was added to each well of the plates. The plates were
incubated at 37.degree. C. incubator (non-humidified) for 120 min.
OD was taken with spectrophotometer at 490 nm.
[0202] FIG. 8 illustrates the survival of adipocytes in sodium
deoxycholate solution and sodium deoxycholate with 1%
phosphatidylcholine solution. The adipocyte cells are primary
(i.e., taken freshly from a person) adipocytes, cultured in vitro
to maturity before treatment with DC. On y-axis, 1 represents 100%
survival when no DC is added. All % concentrations are w/v %. LD50
(concentration at which 50% of the fat cells are dead) of only
sodium deoxycholate solution for adipoocyte cell line was found to
be around 0.02% sodium deoxycholate. The addition of sodium
deoxycholate with 1% phosphatidylcholine solution increases the
LD50 by .about.5-6 fold.
[0203] Results of this study demonstrate that addition of 1%
phosphatidylcholine inhibits apoptosis in vitro i.e., the presence
of PC makes it 5-times harder for DC to kill the adipocytes,
showing that PC detracts from, and thus does not enhance the fat
cell killing power of DC. TABLE-US-00005 TABLE 5 Adipocytes after
treatment with DC only and DC + 1% PC Adipocytes surviving
Percentage Adipocytes Percentage in DC Aver- surviving surviving in
Aver- surviving DC only age % DC + 1% PC age % 0% 0.036 0.113 1
0.191 0.200 1 0.112 0.19 0.114 0.22 0.005 0.52 0.111 0.982 0.195
0.186 0.930 0.105 0.178 0.117 0.186 0.01 0.067 0.131 1.163 0.174
0.178 0.891 0.133 0.183 0.13 0.179 0.05 0.01 0.09 0.079 0.167 0.159
0.795 0.009 0.164 0.008 0.147 0.1 0 0 0 0.138 0.141 0.707 0 0.14 0
0.147 0.5 0 0 0 0.032 0.024 0.123 0 0.02 0 0.022
Example 8
Effects of Addition of Phosphatidylcholine to 4.75% Sodium
Deoxycholate Solutions on Viable Adipocytes
[0204] FIG. 9 illustrates survival of adipocytes when
phosphatidylcholine is added to 4.75% sodium deoxycholate solution.
On y-axis, 1 represents 100% survival. All % concentrations are w/v
%. Survival of the cells in the absence of DC and PC is on an
average 0.113.
[0205] Results of this study demonstrate that addition of
phosphatidylcholine to sodium deoxycholate solution does not
necessarily contribute to adipolysis in vitro, as deoxycholate is
both necessary and sufficient to mediate 100% adipolysis of
cultured adipocytes. Furthermore, a close examination of these data
(Table 6) shows that 5% phosphatidylcholine actually inhibits
adipolysis slightly, which is not surprising in the light of the
5-6 fold increase in LD50 conferred by the addition of
phosphatidylcholine (as demonstrated in example 7). These data
collectively support the notion that sodium deoxycholate is an
active ingredient for adipolysis, and that phosphatidylcholine is
not only unnecessary, but actually inhibits fat removal.
TABLE-US-00006 TABLE 6 Adipocytes after addition of PC to 4.75% DC
Adipocytes surviving by Percentage PC adding PC to 4.75% DC Average
surviving % 0 0 0 0 0 0 0.01 0 0 0 0 0 0.1 0 0 0 0 0 1 0 0 0 0 0 5
0.002 0.009 0.079 0.013 0.012
Example 9
Inhibition of Adipolysis by Pre-Incubation with Human Lipoma
Fat
[0206] The experiment was performed on a series of immortalized
melanocyte cell lines. Resected human lipomas were ground up, and
DC-containing media was mixed with the human fat and agitated for
24 hours. The material was centrifuged to pellet insoluble
contents. The supernatants were then added on top of cultured
melanocytes. Control was the identical DC-containing media not
exposed to the fat chunks. Also, fat alone (no media) was added as
a control.
[0207] FIG. 10 shows that pre-incubation of 0.1% DC with the fat
(Table 7) appears to reduce killing. It could be by releasing some
form of inhibitor (e.g., example 7 shows that PC inhibits DC
killing in vitro) or could be by retaining the DC in the fat pellet
that was spun out in the centrifuge. But it is evident in this
experiment that the presence of the fat limits the killing
properties of DC, may be by direct inhibition (like PC) or by
sequestration (i.e., into the fat pellet). This can further explain
the observation that injection of DC into fat spares the
surrounding tissues. TABLE-US-00007 TABLE 7 Inhibition of
adipolysis by pre-incubation with human lipoma fat Control DC 0.1%
DC 0.1% + fat Fat A375M 0.598 0.602 0.616 0.066 0.063 0.063 0.456
0.382 0.365 0.494 0.517 0.601 Actual 0.598 0.602 0.616 0.066 0.063
0.063 0.456 0.382 0.365 0.494 0.517 0.601 Average 0.605 0.064 0.401
0.537 Percentage 1.000 0.106 0.662 0.888 CHL 0.63 0.683 0.663 0.064
0.066 0.064 0.402 0.426 0.378 0.612 0.623 0.646 Actual 0.63 0.683
0.663 0.064 0.066 0.064 0.402 0.426 0.378 0.612 0.623 0.646 Average
0.659 0.065 0.402 0.627 Percentage 1 0.098 0.610 0.952 WM266 0.548
0.56 0.567 0.07 0.061 0.058 0.281 0.308 0.314 0.48 0.507 0.533
Actual 0.548 0.56 0.567 0.07 0.061 0.058 0.281 0.308 0.314 0.48
0.507 0.533 Average 0.558 0.063 0.301 0.507 Percentage 1.000 0.113
0.539 0.907 SKmel28 0.583 0.643 0.620 0.061 0.060 0.060 0.290 0.304
0.302 0.569 0.591 0.616 Actual 0.583 0.643 0.620 0.061 0.060 0.060
0.290 0.304 0.302 0.569 0.591 0.616 Average 0.615 0.060 0.299 0.592
Percentage 1.000 0.098 0.485 0.962 Blank 0.052 0.052 0.052 0.052
0.052 0.052 0.052 0.052 0.052 0.052 0.052 0.052
Example 10
Human Lipoma Studies with Sodium Deoxycholate Compositions
[0208] Six patients having 12 lipomas were injected with sodium
deoxycholate (DC) solutions over a period of six months. All
injected lipomas reduced in size after at least one treatment
(Table 8). Measurements were made by physical measurements and
ultrasound imaging. The results of this study demonstrate that the
detergent effects of deoxycholate seen on fat in animal tissues are
reproducible clinically in humans and that there is a significant
reduction in the size of lipomas after the treatment.
TABLE-US-00008 TABLE 8 Human lipoma studies with sodium
deoxvcholate compositions Pre-treatment Post-treatment Change in
Total Lipoma size (cm) size volume (%) Treatments 1 1 0 -100% 1 2 2
1.0, 0.3, 0.5 n/a 3 3 2 .times. 2.5 1 .times. 1.5 84% 3 4 4 .times.
3.5 2 84% 4 5 2 .times. 1.8 1.3 92% 4 6 2.8 0.5 99% 1 7 3 .times. 2
0.8 97% 2 8 1 0 100% 2 9 2 .times. 1 1.2 .times. 0.8 67% 2 10 2 1.3
73% 2 11 2 1.4 .times. 1 79% 1 12 1 0.8 50% 1
[0209] Unless otherwise indicated, all numbers expressing
quantities of ingredients, properties such as molecular weight,
reaction conditions, and so forth used in the specification and
claims are to be understood as being modified in all instances by
the term "about." Accordingly, unless indicated to the contrary,
the numerical parameters set forth in the following specification
and attached claims are approximations that may vary depending upon
the desired properties sought to be obtained by the present
invention. At the very least, and not as an attempt to limit the
application of the doctrine of equivalents to the scope of the
claims, each numerical parameter should at least be construed in
light of the number of reported significant digits and by applying
ordinary rounding techniques. Notwithstanding that the numerical
ranges and parameters setting forth the broad scope of the
invention are approximations, the numerical values Size (cm) set
forth in the specific examples are reported as precisely as
possible. Any numerical value, however, inherently contains certain
errors necessarily resulting from the standard deviation found in
their respective testing measurements.
[0210] The terms "a" and "an" and "the" and similar referents used
in the context of describing the invention (especially in the
context of the following claims) are to be construed to cover both
the singular and the plural, unless otherwise indicated herein or
clearly contradicted by context. Recitation of ranges of values
herein is merely intended to serve as a shorthand method of
referring individually to each separate value falling within the
range. Unless otherwise indicated herein, each individual value is
incorporated into the specification as if it were individually
recited herein. All methods described herein can be performed in
any suitable order unless otherwise indicated herein or otherwise
clearly contradicted by context. The use of any and all examples,
or exemplary language (e.g., "such as") provided herein is intended
merely to better illuminate the invention and does not pose a
limitation on the scope of the invention otherwise claimed. No
language in the specification should be construed as indicating any
non-claimed element essential to the practice of the invention.
[0211] Groupings of alternative elements or embodiments of the
invention disclosed herein are not to be construed as limitations.
Each group member may be referred to and claimed individually or in
any combination with other members of the group or other elements
found herein. It is anticipated that one or more members of a group
may be included in, or deleted from, a group for reasons of
convenience and/or patentability. When any such inclusion or
deletion occurs, the specification is herein deemed to contain the
group as modified thus fulfilling the written description of all
Markush groups used in the appended claims.
[0212] Preferred embodiments of this invention are described
herein, including the best mode known to the inventors for carrying
out the invention. Of course, variations on those preferred
embodiments will become apparent to those of ordinary skill in the
art upon reading the foregoing description. The inventor expects
skilled artisans to employ such variations as appropriate, and the
inventors intend for the invention to be practiced otherwise than
specifically described herein. Accordingly, this invention includes
all modifications and equivalents of the subject matter recited in
the claims appended hereto as permitted by applicable law.
Moreover, any combination of the above-described elements in all
possible variations thereof is encompassed by the invention unless
otherwise indicated herein or otherwise clearly contradicted by
context.
[0213] Furthermore, numerous references have been made to patents
and printed publications throughout this specification. Each of the
above cited references and printed publications are herein
individually incorporated by reference in their entirety.
[0214] In closing, it is to be understood that the embodiments of
the invention disclosed herein are illustrative of the principles
of the present invention. Other modifications that may be employed
are within the scope of the invention. Thus, by way of example, but
not of limitation, alternative configurations of the present
invention may be utilized in accordance with the teachings herein.
Accordingly, the present invention is not limited to that precisely
as shown and described.
* * * * *