U.S. patent application number 13/605767 was filed with the patent office on 2013-01-24 for methods for inhibiting preterm labor and uterine contractility disorders and preventing cervical ripening.
This patent application is currently assigned to DIGNITY HEALTH. The applicant listed for this patent is Robert Garfield, Leili Shi, Shao-Qing Shi. Invention is credited to Robert Garfield, Leili Shi, Shao-Qing Shi.
Application Number | 20130023505 13/605767 |
Document ID | / |
Family ID | 44563831 |
Filed Date | 2013-01-24 |
United States Patent
Application |
20130023505 |
Kind Code |
A1 |
Garfield; Robert ; et
al. |
January 24, 2013 |
METHODS FOR INHIBITING PRETERM LABOR AND UTERINE CONTRACTILITY
DISORDERS AND PREVENTING CERVICAL RIPENING
Abstract
The invention relates to methods and pharmaceutical compositions
for inhibiting or preventing preterm birth, inhibiting or delaying
cervical ripening, inhibiting myometrial contractility and treating
or inhibiting uterine contractility disorders. The methods comprise
administering an effective amount of a composition comprising
steroid hormones such as soluble progesterone.
Inventors: |
Garfield; Robert; (Goodyear,
AZ) ; Shi; Shao-Qing; (Goodyear, AZ) ; Shi;
Leili; (Goodyear, AZ) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Garfield; Robert
Shi; Shao-Qing
Shi; Leili |
Goodyear
Goodyear
Goodyear |
AZ
AZ
AZ |
US
US
US |
|
|
Assignee: |
DIGNITY HEALTH
Phoenix
AZ
|
Family ID: |
44563831 |
Appl. No.: |
13/605767 |
Filed: |
September 6, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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PCT/US11/27788 |
Mar 9, 2011 |
|
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13605767 |
|
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61311944 |
Mar 9, 2010 |
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61434309 |
Jan 19, 2011 |
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Current U.S.
Class: |
514/178 |
Current CPC
Class: |
A61P 15/06 20180101;
A61K 31/405 20130101; A61K 31/57 20130101; A61K 31/4422 20130101;
A61K 9/0043 20130101; A61K 9/0014 20130101; A61K 9/0034 20130101;
A61K 47/44 20130101; A61K 9/0019 20130101; A61K 31/57 20130101;
A61K 2300/00 20130101; A61K 31/4422 20130101; A61K 2300/00
20130101; A61K 31/405 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/178 |
International
Class: |
A61K 31/567 20060101
A61K031/567; A61P 15/06 20060101 A61P015/06 |
Claims
1. A method for inhibiting preterm birth in a subject in need
thereof comprising: (i) providing a composition comprising a
steroid hormone or a pharmaceutical equivalent, analog, derivative
or a salt thereof, which inhibits preterm birth; and (ii)
administering a therapeutically effective amount of the composition
to the subject to inhibit preterm birth, thereby inhibiting preterm
birth.
2. A method for preventing preterm birth in a subject in need
thereof by the method of claim 1.
3. A method for delaying cervical ripening in a subject in need
thereof comprising: (i) providing a composition comprising a
steroid hormone or a pharmaceutical equivalent, analog, derivative
or a salt thereof, which delays cervical ripening; and (ii)
administering a therapeutically effective amount of the composition
to the subject to inhibit cervical ripening, thereby inhibiting
cervical ripening.
4. A method for inhibiting cervical ripening in a subject in need
thereof by the method of claim 3.
5. The method of claim 1, wherein the composition further comprises
an effective amount of an agent to render the steroid hormone
soluble.
6. The method of claim 5, wherein the agent is any one or more of
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil saline solution and/or
glucose solution.
7. The method of claim 6, wherein the agent is fish oil, peppermint
oil or saline solution.
8. The method of claim 5, wherein the effective amount of the agent
is about 0.05-0.1 ml/mg of steroid hormone, 0.1-0.2 ml/mg of
steroid hormone, 0.2-0.3 ml/mg of steroid hormone, 0.3-0.4 ml/mg of
steroid hormone, 0.4-0.5 ml/mg of steroid hormone, 0.5-0.6 ml/mg of
steroid hormone, 0.6-0.7 ml/mg of steroid hormone, 0.7-0.8 ml/mg of
steroid hormone, 0.8-0.9 ml/mg of steroid hormone, 0.9-1.0 ml/mg of
steroid hormone, 1.0-5.0 ml/mg of steroid hormone, 5.0-10.0 ml/mg
of steroid hormone, 10.0-15.0 ml/mg of steroid hormone, 15.0-20.0
ml/mg of steroid hormone, 20.0-25.0 ml/mg of steroid hormone or
25.0-30.0 ml/mg of steroid hormone.
9. The method of claim 1, wherein the composition is administered
beginning at about 18.sup.th to about 22.sup.nd of gestation and
ending at about 37.sup.th week of gestation.
10. The method of claim 1, wherein the composition is administered
beginning at about 16.sup.th of gestation and ending at about
37.sup.th week of gestation.
11. The method of claim 1, wherein the composition is administered
beginning at the time of positive pregnancy until the 37.sup.th
week of gestation or from time preterm labor is suspected to a time
when delivery is imminent.
12. The method of claim 1, wherein the composition is administered
for about 2 to 4 weeks, for about 4 to 6 weeks, for about 6 to 8
weeks, for about 8 to 10 weeks, for about 10 to 12 weeks, for about
12 to 14 weeks or for about 14 to 19 weeks.
13. The method of claim 1, wherein the composition is administered
for about 20 weeks, for about 21 weeks, for about 22 weeks, for
about 23 weeks, for about 25 weeks, for about 26 weeks, for about
27 weeks, for about 28 weeks or for about 29 weeks.
14. The method of claim 1, wherein the steroid hormone is a
progestogen or a pharmaceutical equivalent, analog, derivative or a
salt thereof.
15. The method of claim 14, wherein the progestogen is a compound
having the formula ##STR00007## or a pharmaceutical equivalent,
analog, derivative or a salt thereof.
16. The method of claim 14, wherein the progestogen is progesterone
(P4) or a pharmaceutical equivalent, analog, derivative or a salt
thereof.
17. The method of claim 14, wherein the progestogen is
17-hydroxyprogesterone (17OHP) or a pharmaceutical equivalent,
analog, derivative or a salt thereof.
18. The method of claim 14, wherein the progestogen is a
progestin.
19. The method of claim 18, wherein the progestin is
17-hydroxyprogesterone caproate (17P) or promegestone (R5020).
20. The method of claim 1, wherein the composition is in a soluble
form, crystalline form, gel form, tablet form or encapsulated
form.
21. The method of claim 1, wherein the composition is administered
subcutaneously, intradermally, intravenously, intramuscularly,
intraperitonealy, orally, vaginally or via inhalation.
22. The method of claim 1, wherein the composition is administered
topically.
23. The method of claim 1, wherein the effective amount of the
steroid hormone is about 0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day,
10-15 mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day, 30-35
mg/day, 35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55 mg/day,
55-60 mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day, 75-80
mg/day, 80-85 mg/day, 85-90 mg/day, 90-95 mg/day, 95-100 mg/day,
100-200 mg/day, 200-300 mg/day, 300-500 mg/day, 500-700 mg/day,
700-1000 mg/day, 1000-2000 mg/day, 2000-3000 mg/day, 3000-4000
mg/day or 4000-5000 mg/day.
24. A method for inhibiting myometrial contractility in a subject
in need thereof comprising: (i) providing a composition comprising
progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof, which inhibit myometrial
contractility; and (ii) administering a therapeutically effective
amount of the composition to the subject to inhibit myometrial
contractility, thereby inhibiting myometrial contractility.
25. A method for treating uterine contractility disorders in a
subject in need thereof by the method of claim 24.
26. A method for inhibiting uterine contractility disorders in a
subject in need thereof by the method of claim 24.
27. The method of claim 24, wherein the composition further
comprises an agent to render the steroid hormone soluble.
28. The method of claim 27, wherein the agent is any one or more of
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil, saline solution and/or
glucose solution.
29. The method of claim 24, wherein the composition further
comprises an effective amount of nifedipine or indomethacin.
30. The method of claim 24, wherein the composition is administered
subcutaneously, intradermally, intravenously, intramuscularly,
intraperitonealy, orally, vaginally or via inhalation.
31. The method of claim 24, wherein the composition is administered
topically.
32. The method of claim 1, wherein the subject is selected from the
group consisting of human, non-human primate, monkey, ape, dog,
cat, cow, horse, rabbit, mouse, pig and rat.
33. The method of claim 27, wherein the subject is human.
34. The method of claim 24, wherein the effective amount of
composition is about 0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day, 10-15
mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day, 30-35 mg/day,
35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55 mg/day, 55-60
mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day, 75-80 mg/day,
80-85 mg/day, 85-90 mg/day, 90-95 mg/day 95-100 mg/day, 100-200
mg/day, 200-300 mg/day, 300-500 mg/day, 500-700 mg/day, 700-1000
mg/day, 1000-2000 mg/day, 2000-3000 mg/day, 3000-4000 mg/day or
4000-5000 mg/day.
35. The method of claim 29, wherein progesterone and nifedipine are
administered concurrently or sequentially.
36. The method of claim 29, wherein progesterone and indomethacin
are administered concurrently or sequentially.
37. The method of claim 29, wherein the effective amount of
nifedipine is about 0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day, 10-15
mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day, 30-35 mg/day,
35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55 mg/day, 55-60
mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day, 75-80 mg/day,
80-85 mg/day, 85-90 mg/day, 90-95 mg/day or 95-100 mg/day.
38. The method of claim 29, wherein the effective amount of
indomethacin is about 0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day, 10-15
mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day, 30-35 mg/day,
35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55 mg/day, 55-60
mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day, 75-80 mg/day,
80-85 mg/day, 85-90 mg/day, 90-95 mg/day or 95-100 mg/day.
39. The method of claim 6, wherein cyclodextrins are selected from
the group consisting of .alpha.-cyclodextrin, .beta.-cyclodextrin,
.gamma.-cyclodextrin, 2-hydroxypropyl-.beta.-cyclodextrin and
methyl-.beta.-cyclodextrin.
40. A pharmaceutical composition comprising a therapeutically
effective amount of a compound of the formula: ##STR00008## or a
pharmaceutical equivalent, derivative, analog, and/or salt thereof,
and a pharmaceutically acceptable carrier.
41. The pharmaceutical composition of claim 40, further comprising
an agent to render the compound soluble.
42. The pharmaceutical composition of claim 41, wherein the agent
is any one or more of cyclodextrins, sesame oil, fish oil, corn
oil, olive oil, coconut oil, krill oil, omega fatty acids, mineral
oil, peppermint oil, flaxseed oil, vitamin E oil, argan oil, saline
solution and/or glucose solution.
43. The pharmaceutical composition of claim 40, further comprising
an effective amount of nifedipine and/or indomethacin.
44. The pharmaceutical composition of claim 41, wherein an
effective amount of the agent is about 0.05-0.1 ml/mg of steroid
hormone, 0.1-0.2 ml/mg of steroid hormone, 0.2-0.3 ml/mg of steroid
hormone, 0.3-0.4 ml/mg of steroid hormone, 0.4-0.5 ml/mg of steroid
hormone, 0.5-0.6 ml/mg of steroid hormone, 0.6-0.7 ml/mg of steroid
hormone, 0.7-0.8 ml/mg of steroid hormone, 0.8-0.9 ml/mg of steroid
hormone, 0.9-1.0 ml/mg of steroid hormone, 1.0-5.0 ml/mg of steroid
hormone, 5.0-10.0 ml/mg of steroid hormone, 10.0-15.0 ml/mg of
steroid hormone, 15.0-20.0 ml/mg of steroid hormone, 20.0-25.0
ml/mg of steroid hormone or 25.0-30.0 ml/mg of steroid hormone.
45. The pharmaceutical composition of claim 43, wherein the
effective amount of indomethacin is about 0.5-1 mg/day, 1-5 mg/day,
5-10 mg/day, 10-15 mg/day, 15-20 mg/day, 20-25 mg/day, 25-30
mg/day, 30-35 mg/day, 35-40 mg/day, 40-45 mg/day, 45-50 mg/day,
50-55 mg/day, 55-60 mg/day, 60-65 mg/day, 65-70 mg/day, 70-75
mg/day, 75-80 mg/day, 80-85 mg/day, 85-90 mg/day, 90-95 mg/day or
95-100 mg/day.
46. The pharmaceutical composition of claim 43, wherein the
effective amount of nifedipine is about 0.5-1 mg/day, 1-5 mg/day,
5-10 mg/day, 10-15 mg/day, 15-20 mg/day, 20-25 mg/day, 25-30
mg/day, 30-35 mg/day, 35-40 mg/day, 40-45 mg/day, 45-50 mg/day,
50-55 mg/day, 55-60 mg/day, 60-65 mg/day, 65-70 mg/day, 70-75
mg/day, 75-80 mg/day, 80-85 mg/day, 85-90 mg/day, 90-95 mg/day or
95-100 mg/day.
47. The pharmaceutical composition of claim 40, wherein the
therapeutically effective amount of the steroid hormone is about
0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day, 10-15 mg/day, 15-20 mg/day,
20-25 mg/day, 25-30 mg/day, 30-35 mg/day, 35-40 mg/day, 40-45
mg/day, 45-50 mg/day, 50-55 mg/day, 55-60 mg/day, 60-65 mg/day,
65-70 mg/day, 70-75 mg/day, 75-80 mg/day, 80-85 mg/day, 85-90
mg/day, 90-95 mg/day, 95-100 mg/day, 100-200 mg/day, 200-300
mg/day, 300-500 mg/day, 500-700 mg/day, 700-1000 mg/day, 1000-2000
mg/day, 2000-3000 mg/day, 3000-4000 mg/day or 4000-5000 mg/day.
48. The pharmaceutical composition of claim 42, wherein the agent
is fish oil, peppermint oil or saline solution.
49. A method of increasing bioavailability of a steroid hormone
comprising: (i) providing a composition comprising a steroid
hormone or a pharmaceutical equivalent, analog, derivative or a
salt thereof; and (ii) administering a therapeutically effective
amount of the composition via topical delivery, thereby increasing
bioavailability of the steroid hormone.
50. The method of claim 49, wherein the therapeutically effective
composition comprises an effective amount of an agent to render the
steroid hormone soluble, the steroid hormone or a pharmaceutical
equivalent, analog, derivative or a salt thereof comprises a
progestogen, and increasing bioavailability comprises higher levels
of the progestogen in blood plasma.
Description
[0001] This application is a continuation-in-part of, and includes
a claim of priority under 35 U.S.C. .sctn.120 to, International
Application No. PCT/US2011/027788, filed on Mar. 9, 2011, which
designated the U.S. and that International Application was
published under PCT Article 21(2) in English. This application also
includes a claim of priority under 35 U.S.C. .sctn.119(e) to U.S.
Ser. No. 61/311,944 filed Mar. 9, 2010 and to U.S. Ser. No.
61/434,309 filed Jan. 19, 2011, the contents of all of which are
herein incorporated by reference.
FIELD OF INVENTION
[0002] The invention relates to methods for inhibiting or
preventing preterm birth, delaying cervical ripening, inhibiting
myometrial contractility and treating uterine contractility
disorders, in subjects in need thereof. The method comprises
administering an effective amount of a steroid hormone such as
progesterone or a pharmaceutical equivalent, analog, derivative or
a salt thereof. The invention also relates to pharmaceutical
compositions and kits, comprising steroid hormones such as
progesterone or a pharmaceutical equivalent, analog, derivative or
a salt thereof and a pharmaceutically acceptable carrier.
BACKGROUND OF THE INVENTION
[0003] All publications herein are 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. The following description includes information that may
be useful in understanding the present invention. It is not an
admission that any of the information provided herein is prior art
or relevant to the presently claimed invention, or that any
publication specifically or implicitly referenced is prior art.
[0004] Preterm birth (less than 37 completed weeks of gestation) is
one of the major problems and challenges in obstetrics. The
frequency of preterm births is about 12-13% in the USA and 5-9% in
many other developed countries. Despite all efforts to reduce the
number of preterm births the problem is continuing to escalate.
Since 1990 the percentage of births delivered preterm has risen
more than 20 percent and is 36 percent higher since the early 1980s
in the USA..sup.3 Preterm birth is not only a major determinant of
neonatal and infant morbidity, including neurodevelopmental
handicaps, chronic respiratory problems, intraventricular
hemorrhage, infection, retrolental fibroplasia, and necrotizing
enterocolitis, but it is also the single most important cause of
perinatal mortality in North America, Europe and particularly in
undeveloped countries. Additionally, the neonatal and long-term
health care costs of preterm infants impose a considerable economic
strain both on individual families and on healthcare costs
(>$26.2 billion in 2005 in the USA).
[0005] Both uterine and cervical functions play important roles in
the onset and progression of term and preterm labor and delivery.
The cervix undergoes dramatic changes throughout pregnancy and
parturition, a process that is termed cervical ripening--from a
firm, rigid and closed state that is protecting the special milieu
of the fetus from the environment, to a soft and easy-to-open state
that is essential for successful vaginal delivery. The cervix is
dominated by fibrous connective tissue that is composed of an
extracellular matrix which consists mostly of collagen (70% type I
and .about.30% type III) with elastin and proteoglycans and a
cellular portion that consists of smooth muscles, fibroblasts,
epithelium and blood vessels. Cervical ripening is an active
biochemical process, which occurs independent of uterine
contractions. Studies have shown that cervical ripening is
associated with a strong reorganization of the extracellular
matrix, especially collagen: Not only does the concentration
decrease by 30-70%, but there is also a switch from insoluble to
more soluble collagen. Ripening of the cervix is an
inflammatory-like reaction with infiltration of leukocytes,
increase of cytokines (interleukin (IL)-1 and IL-8) and an increase
in metalloproteinases. This process also seems to be at least
partially regulated by steroid hormones (in particular progesterone
(P4) and estrogen), as antiprogestins successfully induce cervical
ripening. Other hormones and mediators shown to be involved in
cervical ripening are dihydrotestosterone, prostaglandins, and
local mediators such as platelet-activating factor and nitric
oxide. Various methods have been used to evaluate cervical ripening
and effects of progestins, including cervical length. However the
biochemical mechanisms that are responsible for the remarkable
changes in the cervix remain poorly understood. Although
progesterone has been known to be used for recurrent or high risk
preterm labor (PTL), its current use is crystalline progesterone in
micronized form, and used to treat preterm labor and uterine
contractile disorders by the often inconvenient and less effective
routes of vaginal, oral or IM. Since the half life of progesterone
is roughly 32 hours, progesterone by this matter must be given
daily, and since crystalline progesterone can only be dissolved in
oil, it can only be applied via vaginal administration or injected
IM.
SUMMARY OF THE INVENTION
[0006] The invention provides methods for inhibiting preterm birth
in a subject in need thereof. The methods comprise providing a
composition comprising a steroid hormone (for example progesterone
(P4) or a pharmaceutical equivalent, analog, derivative or a salt
thereof) and administering a therapeutically effective amount of
the composition to the subject to inhibit preterm birth, thereby
inhibiting preterm birth.
[0007] The invention also provides methods for preventing preterm
birth in a subject in need thereof. The methods comprise providing
a composition comprising a steroid hormone (for example
progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof) and administering a therapeutically
effective amount of the composition to the subject to prevent
preterm birth, thereby preventing preterm birth.
[0008] The invention further provides methods for inhibiting
cervical ripening in a subject in need thereof. The method
comprises providing a composition comprising a steroid hormone (for
example progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof) and administering a therapeutically
effective amount of the composition to the subject to inhibit
cervical ripening, thereby inhibiting cervical ripening.
[0009] Methods for delaying cervical ripening in a subject in need
thereof are also provided herein. The methods comprise providing a
composition comprising a steroid hormone (for example progesterone
(P4) or a pharmaceutical equivalent, analog, derivative or a salt
thereof) and administering a therapeutically effective amount of
the composition to the subject to delay cervical ripening, thereby
delaying cervical ripening.
[0010] Further provided are methods for inhibiting myometrial
contractility in a subject in need thereof. The methods comprise
providing a composition comprising progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof and
administering a therapeutically effective amount of the composition
to the subject to inhibit myometrial contractility, thereby
inhibiting myometrial contractility.
[0011] The invention also provides methods for treating uterine
contractility disorders in a subject in need thereof. The methods
comprise providing a composition comprising progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof and
administering a therapeutically effective amount of the composition
to the subject to treat uterine contractility disorders, thereby
treating uterine contractility disorders.
[0012] The invention further provides methods for inhibiting
uterine contractility disorders in a subject in need thereof. The
method comprises providing a composition comprising progesterone
(P4) or a pharmaceutical equivalent, analog, derivative or a salt
thereof and administering a therapeutically effective amount of the
composition to the subject to inhibit uterine contractility
disorders, thereby inhibiting uterine contractility disorders.
[0013] The invention also provides that steroid hormones such as
progesterone (P4) may be rendered soluble by mixing with agents
such as cyclodextrins, sesame oil, fish oil, corn oil, olive oil,
coconut oil, krill oil, omega fatty acids, mineral oil, peppermint
oil, flaxseed oil, vitamin E oil, argan oil, saline solution and/or
glucose solution. The composition may be applied topically,
intravenously, subcutaneously or nasally.
[0014] Also provided are pharmaceutical compositions and kits
comprising steroid hormones (for example progesterone or a
pharmaceutical equivalent, analog, derivative or a salt thereof)
and a pharmaceutically acceptable carrier. The pharmaceutical
compositions may be mixed with agents such as cyclodextrins, sesame
oil, fish oil, corn oil, olive oil, coconut oil, krill oil, omega
fatty acids, mineral oil, peppermint oil, flaxseed oil, vitamin E
oil, argan oil, saline solution and/or glucose solution, to render
the steroid hormone soluble.
BRIEF DESCRIPTION OF THE FIGURES
[0015] Exemplary embodiments are illustrated in referenced figures.
It is intended that the embodiments and figures disclosed herein
are to be considered illustrative rather than restrictive.
[0016] FIG. 1 depicts bar graphs showing means.+-.SD of cervical
light-induced fluorescence (LIF) obtained in vivo from nonpregnant
(N=3), pregnant (d13, 15 and 17: N=12/group; d19 and d21:
N=11/group; d22: N=6) and postpartum rats (pp 3, 5 and 7: N=7; pp
10: N=6). Significant differences (P<0.05) between groups are
marked with different letters.
[0017] FIG. 2 depicts bar graphs showing means.+-.SD of cervical
light-induced fluorescence (LIF) obtained in vivo from pregnant
rats at different days of pregnancy and postpartum (N=6/group)
treated with various progestins or vehicle. FIG. 2A: Daily
treatment with vehicle (controls) or P4 (4 mg, subcutaneously).
Note that delivery is inhibited in the treatment group. FIG. 2B:
Twice a day treatment with vehicle (controls) or vaginal P4 (15 mg
bid). Note that no significant differences are observed at any time
between controls vs. treated animals. FIG. 2C: Treatment daily with
vehicle (controls) or 17P (10 mg, subcutaneously). Note that
significant differences are only observed until day 19 of
gestation. FIG. 2D: Twice a day treatment with vehicle (controls)
or vaginal promegestone (R5020) (1 mg bid). Note that significant
differences are observed only until day 19, but delivery is blocked
in the treatment group. Asterisks indicate P<0.05 compared with
controls.
[0018] FIG. 3 depicts bar graphs showing means.+-.SD of cervical
light-induced fluorescence (LIF) obtained in vivo from pregnant
rats at different days of pregnancy and postpartum (N=6/group)
treated once on day 16 with vehicle (controls) or RU-486 (3 mg
subcutaneously). Asterisks indicate P<0.05 compared with
controls.
[0019] FIG. 4 depicts the percent of animals delivering versus day
of pregnancy after various treatments. FIG. 4A: Delivery times
after daily treatment with vehicle (subcutaneously, controls), P4
(4 mg, subcutaneously) and 17P (10 mg, subcutaneously). Note that
injections of P4 completely blocked delivery, whereas 17P had no
significant effect on delaying term delivery (see also FIG. 5).
FIG. 4B: Percent of animals delivering versus time of delivery
following twice a day treatment with vaginal vehicle (controls),
vaginal P4 (15 mg bid) and vaginal promegestone (R5020) (1 mg bid).
Note that vaginal promegestone (R5020) completely blocked delivery,
whereas vaginal P4 had no significant effect on delaying term
delivery (P>0.05 compared with controls, see also FIG. 5).
[0020] FIG. 5 depicts the time of delivery (=hours after 8 a.m. of
day 22 of gestation) of pregnant rats treated with vehicles
(controls) and various progestins by different routes of
administration--injections (subcutaneously; daily): vehicle: sesame
oil; P4 (4 mg); 17P (10 mg); vaginal (bid): vehicle: Replens; P4
(15 mg, Crinone); promegestone (R5020) (1 mg); oral (bid): vehicle:
sesame oil or H.sub.2O; P4 (15 mg); topical (bid): vehicle:
Replens, sesame oil or fish oil; P4 (15 mg). Rats with delayed
parturition were sacrificed on day 25. Asterisks indicate P<0.05
compared with controls.
[0021] FIG. 6 depicts plasma levels of progesterone following
various treatments. Plasma progesterone (P4) levels in pregnant
rats at day 18 and 21 days of gestation after treatment with P4
vaginal gel (15 mg bid), P4 injections (4 mg subcutaneous daily),
topical P4 in fish oil (15 mg bid) and controls (Ctr) treated with
vehicles.
[0022] FIG. 7 depicts, in accordance with an embodiment herein, an
organ chamber system set up for study of uterine contractions.
[0023] FIG. 8 depicts, in accordance with an embodiment herein,
effects of progesterone (depicted as PR) on human myometrial
contractions. As shown, 10.sup.-4M progesterone inhibited
myometrial contractions.
[0024] FIG. 9 depicts, in accordance with an embodiment herein,
effects of soluble (SOL) progesterone and crystalline P4 in ethanol
(ET) on myometrial contractility.
[0025] FIG. 10 depicts, in accordance with an embodiment herein,
effects of various concentrations of MgSO.sub.4 on myometrial
contractility.
[0026] FIG. 11 depicts, in accordance with an embodiment herein,
effects of progesterone (P4) with MgSO.sub.4 on myometrial
contractility.
[0027] FIG. 12 depicts, in accordance with an embodiment herein,
effects of nifedipine (Nife) and progesterone (P4) and their
combination on myometrial contractility.
[0028] FIG. 13 depicts, in accordance with an embodiment herein,
effects of indomethacin (IND) and progesterone (P4) and their
combination on myometrial contractility.
[0029] FIG. 14 depicts, in accordance with an embodiment herein,
effect of pinacidil (10.sup.-6.5 M) alone, or with P4 (10.sup.-5M)
on human myometrium contractility (term, not in labor).
[0030] FIG. 15 depicts, in accordance with an embodiment herein, in
vivo electromyographic activity of uterine contractility over one
hour periods after treatment with soluble progesterone (P4)
(dissolved in 0.9% NaCl) at 2 mg/ml marked in rats.
[0031] FIG. 16 depicts, in accordance with an embodiment herein, in
vivo electromyographic activity of uterine contractility over four
minute periods after treatment with soluble progesterone (P4)
(dissolved in 0.9% NaCl) at 2 mg/ml marked in rats.
[0032] FIG. 17 depicts, in accordance with an embodiment herein,
electromyographic activity of uterine contractility in rats both
(A) before treatment, and (B) after treatment, with soluble
progesterone (P4) (dissolved in 0.9% NaCl) at 2 mg/ml.
[0033] FIG. 18 depicts effects of various routes of administration
of progesterone (P4) in delivery of pregnant rats treated beginning
at day 13 (D13) of gestation.
[0034] FIG. 19 depicts delivery time after treatment beginning at
day 19 (D19) of gestation with progesterone (P4) with various oil
vehicles.
[0035] FIG. 20 depicts delay in pregnant rats treated with
progesterone (P4) beginning at day 20 (D20) of gestation with
various fish oil vehicles.
[0036] FIG. 21 depicts delay of delivery in pregnant rats treated
with progesterone (P4) beginning at day 20 (D20) of gestation with
various vehicles.
[0037] FIG. 22 depicts delay of delivery in pregnant rats treated
with progesterone (P4) either subcutaneously or topically beginning
at days 19, 20, 21 or 22 of gestation.
[0038] FIG. 23 depicts chemical structures of some embodiments of
the invention.
[0039] FIG. 24 depicts pharmokinetic data related to plasma serum
levels after topical delivery in rats following various treatments
on day 19 of gestation (single treatment only).
DETAILED DESCRIPTION OF THE INVENTION
[0040] Unless defined otherwise, technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs.
Singleton et al., Dictionary of Microbiology and Molecular Biology
3.sup.rd ed., J. Wiley & Sons (New York, N.Y. 2001); March,
Advanced Organic Chemistry Reactions, Mechanisms and Structure
5.sup.th ed., J. Wiley & Sons (New York, N.Y. 2001); and
Sambrook and Russel, Molecular Cloning: A Laboratory Manual 3rd
ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor, N.Y.
2001), provide one skilled in the art with a general guide to many
of the terms used in the present application.
[0041] One skilled in the art will recognize many methods and
materials similar or equivalent to those described herein, which
could be used in the practice of the present invention. Indeed, the
present invention is in no way limited to the methods and materials
described. For purposes of the present invention, the following
terms are defined below.
[0042] "Treatment" and "treating," as used herein refer to both
therapeutic treatment and prophylactic or preventative measures,
wherein the object is to prevent or slow down (lessen) the targeted
pathologic condition, prevent the pathologic condition, pursue or
obtain beneficial results, or lower the chances of the individual
developing the condition even if the treatment is ultimately
unsuccessful. Those in need of treatment include those already with
the condition as well as those prone to have the condition or those
in whom the condition is to be prevented.
[0043] As used herein, the term "P4" means progesterone.
[0044] As used herein, progesterone (P4) has the formula (Formula
1):
##STR00001##
[0045] As used herein, Promegestone (R5020) has the formula
(Formula 2):
##STR00002##
[0046] As used herein, 17-hydroxyprogesterone has the formula
(Formula 3):
##STR00003##
[0047] As used herein, 17-hydroxyprogesterone caproate has the
formula (Formula 4):
##STR00004##
[0048] As used herein, Nifedipine has the formula (Formula 5):
##STR00005##
[0049] As used herein, Indomethacin has the formula (Formula
6):
##STR00006##
[0050] As used herein, the term "17P" means
17-alpha-hydroxyprogesterone caproate, a synthetic caproate ester
of the naturally occurring metabolite of progesterone.
[0051] As used herein, the term "LIP" means light-induced
fluorescence.
[0052] As used herein, soluble progesterone is progesterone (P4)
mixed in or suspended in agents that render the progesterone
soluble. Examples of such agents include but are not limited to
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil, saline solution, and/or
glucose solution. For example, encapsulated progesterone (such as
progesterone encapsulated in cyclodextrins) may be solubilized in
any water based solutions including but not limited to saline
solution or glucose solution to render it soluble. Alternately,
powdered progesterone may be may be solubilized in any water based
solutions including but not limited to saline solution or glucose
solution to render it soluble. Additionally, crystalline or
microcrystalline progesterone may be dissolved in various oils to
render it soluble. Saline solution may be isotonic. Saline solution
may be 0.9% w/v NaCl. Any form of progesterone (powdered,
desiccated, crystalline or microcrystalline) may be dissolved with
any suitable agent to render the progesterone soluble.
Therapeutic Methods of the Invention
[0053] The present invention is based, at least in part, on the
findings described herein and thus the present invention describes
methods, pharmaceutical compositons and kits for using steroid
hormones to treat conditions in a subject, such as preterm birth.
In an embodiment of the invention claimed herein, the steroid
hormone is soluble. For example, the steroid hormone may be a
progestogen or a pharmaceutical equivalent, analog, derivative or a
salt thereof. In a preferred embodiment, the progestogen is
progesterone (P4) and is mixed with or suspended in agents that
render the progesterone soluble. These agents include but are not
limited to cyclodextrins, sesame oil, fish oil, corn oil, olive
oil, coconut oil, krill oil, omega fatty acids, mineral oil,
peppermint oil, flaxseed oil, vitamin E oil, argan oil, saline
solution and/or glucose solution. In the most preferred embodiment,
progesterone is mixed with fish oil, peppermint oil or isotonic
saline solution. While not wishing to be bound by any particular
theory, the inventors believe that the combination of progesterone
and agents such as fish oil or peppermint oil, when administered
topically (for example placed or rubbed on the abdominal surface of
pregnant patients), delay the onset of contractions, labor and
delivery. In an embodiment, delay of the delivery acts on the
subject's myometrium, for instance, via inhibition of uterine
contractility.
[0054] The present invention provides methods for inhibiting
preterm birth or preventing preterm birth in a subject in need
thereof. The method comprises providing a composition comprising a
steroid hormone and administering a therapeutically effective
amount of the composition to the subject to so as to inhibit
preterm birth or prevent preterm birth. In an embodiment, the
steroid hormone is soluble and is a progestogen (for example
progesterone) or a pharmaceutical equivalent, analog, derivative or
a salt thereof.
[0055] The invention also provides methods for delaying cervical
ripening or inhibiting cervical ripening in subjects in need
thereof. The methods comprise providing a composition comprising a
steroid hormone and administering a therapeutically effective
amount of the composition to the subject so as to delaying cervical
ripening or inhibit cervical ripening. In an embodiment, the
steroid hormone is soluble and is a progestogen (for example
progesterone) or a pharmaceutical equivalent, analog, derivative or
a salt thereof.
[0056] Also provided herein is a method for inhibiting myometrial
contractility in a subject in need thereof. The method comprises
providing a composition comprising progestogen (for example
progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof) and administering a therapeutically
effective amount of the composition to the subject to inhibit
myometrial contractility. In one embodiment, the progestogen is
soluble.
[0057] Further, the invention provides methods for treating uterine
contractility disorders or inhibiting uterine contractility
disorders in a subject in need thereof. The methods comprise
providing a composition comprising progestogen (for example
progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof) and administering a therapeutically
effective amount of the composition to the subject to treat uterine
contractility disorders or to inhibit uterine contractility
disorders. In one embodiment, inhibiting uterine contractility
disorders is promoting prophylaxis of uterine contractility
disorders. In another embodiment, the progestogen is soluble.
[0058] In one embodiment, the present invention provides a method
of suppressing delivery in a subject comprising administering a
therapeutically effective amount of a composition comprising
progestogen (for example, progesterone, or pharmaceutical
equivalent, analog, derivative, or salt thereof) to the
subject.
[0059] In some embodiments of the claimed methods, the steroid
hormones are progestogens. In a further embodiment, progestogens
include but are not limited to progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof,
17-hydroxyprogesterone or a pharmaceutical equivalent, analog,
derivative or a salt thereof and progestins. In a preferred
embodiment of the invention, the progestogen is progesterone (P4)
or a pharmaceutical equivalent, analog, derivative or a salt
thereof. In another embodiment of the invention, the progestins
include but are not limited to 17-hydroxyprogesterone caproate or a
pharmaceutical equivalent, analog, derivative or a salt thereof or
promegestone (R5020) or a pharmaceutical equivalent, analog,
derivative or a salt thereof. In a further embodiment of the
instant invention, progestins include but are not limited to
medroxyprogesterone acetate, norethindrone, norethindrone acetate,
norethindrone enanthate, desogestrel, levonorgestrel, lynestrenol,
ethynodiol diacetate, norgestrel, norgestimate, norethynodrel,
gestodene, drospirenone, trimegstone, levodesogestrel, gestodyne,
nesterone, etonogestrel and derivatives of 19-nor-testerone.
[0060] Since steroid hormones (such as progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or salts thereof) have low solubilities, these hormones
may be suspended in or mixed with agents that render the steroid
hormones soluble. For instance, suspending or mixing steroid
hormones with agents such as cyclodextrins, sesame oil, fish oil,
corn oil, olive oil, coconut oil, krill oil, omega fatty acids,
mineral oil, peppermint oil, flaxseed oil, vitamin E oil, argan
oil, saline solution and/or glucose solution, facilitates
dissolution. In an embodiment, progesterone (P4) or pharmaceutical
equivalents, analogs, derivatives or salts thereof may be suspended
in or mixed with REPLENS vaginal moisturizer (available from Lil'
Drug Store Products, Inc.).
[0061] In another embodiment, the steroid hormones may be mixed
with carrier molecules such as cyclodextrins to render the steroid
hormone (such as progesterone (P4)) soluble. Examples of
cyclodextrins include but are not limited to .alpha.-cyclodextrin,
.beta.-cyclodextrin, .gamma.-cyclodextrin,
2-hydroxypropyl-.beta.-cyclodextrin and methyl-.beta.-cyclodextrin.
For example by encapsulating progesterone in cyclodextrins and
solubilizing it in any of the agents described above, renders the
progesterone soluble and can be used intravenously, topically,
parenterally, nasally, subcutaneously, intravascularly, vaginally
and/or topically or by other routes of administration.
[0062] In some preferred embodiments, progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof, is
suspended in or mixed with omega fatty acid, omega-3-fatty acids,
fish oil or peppermint oil and may be used topically, parenterally,
nasally or by other routes of administration. In a preferred
embodiment, administration of progesterone mixed with fish oil or
peppermint oil is topical. For example, topical application of
progesterone mixed in fish oil can inhibit delivery (such as
preterm delivery) in a subject in need thereof.
[0063] In another preferred embodiment, progesterone (P4) is
suspended in or mixed with saline solution (for example isotonic
saline solution) to solubilize it and is subsequently administered
via numerous routes of administration including but not limited to
intravenous, topical, parenteral, nasal, subcutaneous injections,
intravascular, vaginal and/or topical or by other routes of
administration.
[0064] In another embodiment, the steroid hormone is
17-hydroxyprogesterone or a pharmaceutical equivalent, analog,
derivative or a salt thereof and may be suspended in or mixed with
agents such as cyclodextrins, sesame oil, fish oil, corn oil, olive
oil, coconut oil, krill oil, omega fatty acids, mineral oil,
peppermint oil, flaxseed oil, vitamin E oil, argan oil, saline
solution and/or glucose solution, to facilitate dissolution.
[0065] In a further embodiment, the steroid hormones are
17-hydroxyprogesterone caproate or a pharmaceutical equivalent,
analog, derivative or a salt thereof and may be suspended in or
mixed with agents such as cyclodextrins, sesame oil, fish oil, corn
oil, olive oil, coconut oil, krill oil, omega fatty acids, mineral
oil, peppermint oil, flaxseed oil, vitamin E oil, argan oil, saline
solution and/or glucose solution, facilitate dissolution.
[0066] In an additional embodiment, the steroid hormone is
promegestone (R5020) or a pharmaceutical equivalent, analog,
derivative or a salt thereof and may be suspended in or mixed
agents such as cyclodextrins, sesame oil, fish oil, corn oil, olive
oil, coconut oil, krill oil, omega fatty acids, mineral oil,
peppermint oil, flaxseed oil, vitamin E oil, argan oil, saline
solution and/or glucose solution, to facilitate dissolution.
[0067] In an embodiment of the invention, the composition of the
claimed methods comprises progesterone (P4) and further comprises
nifedipine, indomethacin and/or oxytocin antagonists (for example
atosiban). In one embodiment, progesterone (P4) and nifedipine may
be administered concurrently. In another embodiment, progesterone
(P4) and nifedipine may be administered sequentially. Similarly, in
one embodiment, progesterone (P4) and indomethacin may be
administered concurrently. In another embodiment, progesterone (P4)
and indomethacin may be administered sequentially. Additionally,
progesterone (P4) and oxytocin antagonists (for example atosiban)
may be administered concurrently. In another embodiment,
progesterone (P4) and oxytocin antagonists (for example atosiban)
may be administered sequentially. In an additional embodiment,
progesterone (P4), nifedipine and indomethacin may be administered
concurrently. Alternatively, progesterone (P4), nifedipine and
indomethacin may be administered sequentially.
[0068] In a preferred embodiment, progesterone (P4) is suspended in
or mixed with agents that render progesterone soluble and is
administered concurrently or sequentially with nifedipine and/or
with indomethacin and/or with oxytocin antagonists such as
atosiban. Examples of agents that render progesterone soluble
include but are not limited to such as cyclodextrins, sesame oil,
fish oil, corn oil, olive oil, coconut oil, krill oil, omega fatty
acids, mineral oil, peppermint oil, flaxseed oil, vitamin E oil,
argan oil, saline solution and/or glucose solution.
[0069] In an embodiment, the composition comprising progesterone
(with or without agents that render the progesterone soluble) and
nifedipine and/or indomethacin is administered topically.
[0070] The subjects treated by the present invention include
mammalian subjects, including but not limited to human, monkey,
ape, dog, cat, cow, horse, goat, pig, rabbit, mouse and rat.
[0071] In a preferred embodiment, the subject is human. In an
embodiment, the human subject is administered a steroid hormone
(for example, progestogen such as progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or a salts thereof) beginning at about the 16.sup.th
week up to the 37.sup.th week of gestation, beginning at about
18.sup.th week up to about 22.sup.nd week of gestation, beginning
at about 18.sup.th week up to about 35.sup.th week of gestation,
beginning at about 18.sup.th week up to about 37.sup.th week of
gestation, beginning at the time of positive pregnancy until the
37.sup.th week of gestation or beginning at the time preterm labor
is suspected up to when time of delivery is imminent. In an
embodiment, the steroid hormone (such as progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or salts thereof) is mixed with or suspended in agents
to render the steroid hormones soluble and is applied topically. In
some preferred embodiments, progesterone (P4) is mixed with fish
oil, peppermint oil or with omega fatty acids and is applied
topically, for example, by placing or rubbing the progesterone
mixed in oil on the abdominal surface of a pregnant woman. In other
preferred embodiments, the steroid hormones such as progesterone
(P4) is mixed with saline solution (such as isotonic saline
solution) and is administered intravenously, topically, nasally or
via any other route of administration.
[0072] In another embodiment, the human subject is administered a
steroid hormone (for example, progestogen such as progesterone
(P4), 17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or a salts thereof) for about 2 to 4 weeks, for about 4
to 6 weeks, for about 6 to 8 weeks, for about 8 to 10 weeks, for
about 10 to 12 weeks, for about 12 to 14 weeks, for about 14 to 19
weeks, for about 20 weeks, for about 21 weeks, for about 22 weeks,
for about 23 weeks, for about 25 weeks, for about 26 weeks, for
about 27 weeks, for about 28 weeks or for about 29 weeks, for about
30 weeks, for about 35 weeks or for about 37 weeks. In an
embodiment, the steroid hormone (such as progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or salts thereof) is mixed with or suspended in agents
to render the steroid hormones soluble and is applied topically. In
some preferred embodiments, progesterone (P4) is mixed with fish
oil, peppermint oil or with omega fatty acids and is applied
topically, for example, by placing or rubbing the progesterone
mixed in oil on the abdominal surface of a pregnant woman. In other
preferred embodiments, the steroid hormones such as progesterone
(P4) is mixed with saline solution (such as isotonic saline
solution) and is administered intravenously, topically, nasally or
via any other route of administration.
[0073] In a further embodiment, the human subject is administered a
steroid hormone (for example, progestogen such as progesterone
(P4), 17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or salts thereof) when the pregnant woman's cervix
length is greater than 1.0 cm, or when the cervix length is less
than or equal to about 3.0 cm, or when the cervix length is between
1.0 and 8.0 cm. In an embodiment, the steroid hormone (such as
progesterone (P4), 17-hydroxyprogesterone, 17-hydroxyprogesterone
caproate, promegestone (R5020), or pharmaceutical equivalents,
analogs, derivatives or salts thereof) is mixed with or suspended
in agents to render the steroid hormones soluble and is applied
topically. In some preferred embodiments, progesterone (P4) is
mixed with fish oil or with omega fatty acids or with cyclodextrins
and is applied topically, for example, by placing or rubbing the
progesterone mixed in oil on the abdominal surface of a pregnant
woman. In other preferred embodiments, the steroid hormones such as
progesterone (P4) is mixed with saline solution (such as isotonic
saline solution) and is administered intravenously, topically,
nasally or via any other route of administration.
[0074] In one embodiment, the steroid hormones in the composition
of the claimed methods (for example, progestogens such as
progesterone (P4), 17-hydroxyprogesterone, 17-hydroxyprogesterone
caproate, promegestone (R5020), or pharmaceutical equivalents,
analogs, derivatives or a salts thereof) are in a soluble form,
crystalline form, gel form, tablet form or encapsulated form.
[0075] Various methods may be utilized to administer the
compositions comprising steroid hormones of the claimed methods,
including but not limited to aerosol, nasal, oral, subcutaneous,
transmucosal, transdermal, parenteral, implantable pump, continuous
infusion, topical application, capsules, injections, intradermally,
intravenously, intramuscularly, intraperitonealy, rectally,
non-vaginally and/or vaginally. In a preferred embodiment of the
claimed invention, the steroid hormone (for example progesterone
(P4)) is administered topically or subcutaneously. In another
preferred embodiment of the claimed methods, promegestone (R5020)
is administered vaginally. In another preferred embodiment,
progesterone (P4) is mixed with or suspended in agents to render it
soluble and is applied topically by placing or rubbing on the
abdominal surface of pregnant patients. Agents that render the
steroid hormones soluble include but are not limited to
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil, saline solution and/or
glucose solution. In the most preferred embodiment of the claimed
invention, progesterone is mixed with fish oil or peppermint oil
and is applied topically to prevent or inhibit preterm birth and/or
delay or inhibit cervical ripening. In other preferred embodiments,
the steroid hormones such as progesterone (P4) is mixed with saline
solution (such as isotonic saline solution) and is administered
intravenously, topically, nasally or via any other route of
administration so as to prevent or inhibit preterm birth and/or
delay or inhibit cervical ripening
[0076] In one embodiment, the present invention provides a method
increasing bioavailability of a therapeutically effective
composition. In one embodiment, increased bioavailability is shown
by higher plasma levels of the composition. In another embodiment,
the therapeutically effective composition includes progestogen (for
example, progesterone, or pharmaceutical equivalent, analog,
derivative, or salt thereof) to the subject. In another embodiment,
the method includes topical delivery. In another embodiment, the
method includes topical delivery in a single treatment.
Dosages of the Invention
[0077] In some embodiments of the invention, the effective amounts
of the steroid hormone (for example progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020)) is about 0.5-1 mg/day, 1-5 mg/day, 5-10
mg/day, 10-15 mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day,
30-35 mg/day, 35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55
mg/day, 55-60 mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day,
75-80 mg/day, 80-85 mg/day, 85-90 mg/day, 90-95 mg/day or 95-100
mg/day, 100-200 mg/day, 200-300 mg/day, 300-400 mg/day, 400-500
mg/day, 500-600 mg/day, 600-700 mg/day, 700-800 mg/day, 800-900
mg/day, 900-1000 mg/day, 1000-1100 mg/day, 1100-1200 mg/day,
1200-1300 mg/day, 1300-1400 mg/day, 1400-1500 mg/day, 1500-1600
mg/day, 1600-1700 mg/day, 1700-1800 mg/day, 1800-1900 mg/day,
1900-2000 mg/day, 2000-2100 mg/day, 2100-2200 mg/day, 2200-2300
mg/day, 2300-2400 mg/day, 2400-2500 mg/day, 2500-2600 mg/day,
2600-2700 mg/day, 2700-2800 mg/day, 2800-2900 mg/day, 2900-3000
mg/day, 3000-3100 mg/day, 3100-3200 mg/day, 3200-3300 mg/day,
3300-3400 mg/day, 3400-3500 mg/day, 3500-3600 mg/day, 3600-3700
mg/day, 3700-3800 mg/day, 3800-3900 mg/day, 3900-4000 mg/day,
4000-4200 mg/day, 4200-4400 mg/day, 4400-4600 mg/day, 4600-4800
mg/day or 4800-5000 mg/day. In one embodiment, the steroid hormone
administered at the aforementioned dosage is progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof. In
another embodiment, the steroid hormone administered at the
aforementioned dosage is 17-hydroxyprogesterone caproate or a
pharmaceutical equivalent, analog, derivative or a salt thereof. In
a further embodiment, the steroid embodiment administered at the
aforementioned dosage is promegestone (R5020) or a pharmaceutical
equivalent, analog, derivative or a salt thereof. In an embodiment,
the steroid hormone, for example progesterone (P4) such as soluble
progesterone (P4) or a pharmaceutical equivalent, analog,
derivative or a salt thereof is administered daily, biweekly,
weekly, every fortnight or monthly. In a preferred embodiment,
progesterone (P4) such as soluble progesterone (P4) or a
pharmaceutical equivalent, analog, derivative or a salt thereof is
administered daily.
[0078] As described above, in one embodiment of the invention the
steroid hormones including but not limited to progesterone (P4),
17-hydroxyprogesterone caproate, promegestone (R5020), or
pharmaceutical equivalents, analogs, derivatives or a salts
thereof, may be suspended in or mixed with agents that render the
steroid hormone soluble. Such agents include but are not limited to
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil, saline solution and/or
glucose solution. The effective amount of the agent may be about
0.05-0.1 ml/mg of steroid hormone, 0.1-0.2 ml/mg of steroid
hormone, 0.2-0.3 ml/mg of steroid hormone, 0.3-0.4 ml/mg of steroid
hormone, 0.4-0.5 ml/mg of steroid hormone, 0.5-0.6 ml/mg of steroid
hormone, 0.6-0.7 ml/mg of steroid hormone, 0.7-0.8 ml/mg of steroid
hormone, 0.8-0.9 ml/mg of steroid hormone, 0.9-1.0 ml/mg of steroid
hormone, 1.0-5.0 ml/mg of steroid hormone, 5.0-10.0 ml/mg of
steroid hormone, 10.0-15.0 ml/mg of steroid hormone, 15.0-20.0
ml/mg of steroid hormone, 20.0-25.0 ml/mg of steroid hormone or
25.0-30.0 ml/mg of steroid hormone. In a preferred embodiment,
progesterone (P4) is mixed with fish oil or peppermint oil or with
saline solution wherein the aforementioned amounts are the
effective amounts of the fish oil or peppermint oil or saline
solution.
[0079] Further, the steroid hormones including but not limited to
progesterone (P4), 17-hydroxyprogesterone caproate, promegestone
(R5020), or pharmaceutical equivalents, analogs, derivatives or a
salts thereof, may be administered concurrently or sequentially
with an effective amount of Nifedipine. In some embodiments of the
invention, the effective amounts of Nifedipine is about 0.5-1
mg/day, 1-5 mg/day, 5-10 mg/day, 10-15 mg/day, 15-20 mg/day, 20-25
mg/day, 25-30 mg/day, 30-35 mg/day, 35-40 mg/day, 40-45 mg/day,
45-50 mg/day, 50-55 mg/day, 55-60 mg/day, 60-65 mg/day, 65-70
mg/day, 70-75 mg/day, 75-80 mg/day, 80-85 mg/day, 85-90 mg/day,
90-95 mg/day or 95-100 mg/day, 100-200 mg/day, 200-300 mg/day,
300-400 mg/day, 400-500 mg/day, 500-600 mg/day, 600-700 mg/day,
700-800 mg/day, 800-900 mg/day, 900-1000 mg/day, 1000-1100 mg/day,
1100-1200 mg/day, 1200-1300 mg/day, 1300-1400 mg/day, 1400-1500
mg/day, 1500-1600 mg/day, 1600-1700 mg/day, 1700-1800 mg/day,
1800-1900 mg/day, 1900-2000 mg/day, 2000-2100 mg/day, 2100-2200
mg/day, 2200-2300 mg/day, 2300-2400 mg/day, 2400-2500 mg/day,
2500-2600 mg/day, 2600-2700 mg/day, 2700-2800 mg/day, 2800-2900
mg/day, 2900-3000 mg/day, 3000-3100 mg/day, 3100-3200 mg/day,
3200-3300 mg/day, 3300-3400 mg/day, 3400-3500 mg/day, 3500-3600
mg/day, 3600-3700 mg/day, 3700-3800 mg/day, 3800-3900 mg/day,
3900-4000 mg/day, 4000-4200 mg/day, 4200-4400 mg/day, 4400-4600
mg/day, 4600-4800 mg/day or 4800-5000 mg/day.
[0080] In another embodiment of the invention, the steroid hormones
including but not limited to progesterone (P4),
17-hydroxyprogesterone caproate, promegestone (R5020), or
pharmaceutical equivalents, analogs, derivatives or a salts
thereof, may be administered concurrently or sequentially with an
effective amount of Indomethacine. In some embodiments of the
invention, the effective amounts of Indomethacin is about 0.5-1
mg/day, 1-5 mg/day, 5-10 mg/day, 10-15 mg/day, 15-20 mg/day, 20-25
mg/day, 25-30 mg/day, 30-35 mg/day, 35-40 mg/day, 40-45 mg/day,
45-50 mg/day, 50-55 mg/day, 55-60 mg/day, 60-65 mg/day, 65-70
mg/day, 70-75 mg/day, 75-80 mg/day, 80-85 mg/day, 85-90 mg/day,
90-95 mg/day or 95-100 mg/day, 100-200 mg/day, 200-300 mg/day,
300-400 mg/day, 400-500 mg/day, 500-600 mg/day, 600-700 mg/day,
700-800 mg/day, 800-900 mg/day, 900-1000 mg/day, 1000-1100 mg/day,
1100-1200 mg/day, 1200-1300 mg/day, 1300-1400 mg/day, 1400-1500
mg/day, 1500-1600 mg/day, 1600-1700 mg/day, 1700-1800 mg/day,
1800-1900 mg/day, 1900-2000 mg/day, 2000-2100 mg/day, 2100-2200
mg/day, 2200-2300 mg/day, 2300-2400 mg/day, 2400-2500 mg/day,
2500-2600 mg/day, 2600-2700 mg/day, 2700-2800 mg/day, 2800-2900
mg/day, 2900-3000 mg/day, 3000-3100 mg/day, 3100-3200 mg/day,
3200-3300 mg/day, 3300-3400 mg/day, 3400-3500 mg/day, 3500-3600
mg/day, 3600-3700 mg/day, 3700-3800 mg/day, 3800-3900 mg/day,
3900-4000 mg/day, 4000-4200 mg/day, 4200-4400 mg/day, 4400-4600
mg/day, 4600-4800 mg/day or 4800-5000 mg/day.
[0081] Typical dosages of an effective amount of a steroid hormone,
progesterone (P4), 17-hydroxyprogesterone, 17-hydroxyprogesterone
caproate, promegestone (R5020), or pharmaceutical equivalents,
analogs, derivatives or a salts thereof, can be in the ranges
recommended by the manufacturer where known therapeutic compounds
are used, and also as indicated to the skilled artisan by the in
vitro responses or responses in animal models. The same or similar
dosing can be used in accordance with various embodiments of the
present invention, or an alternate dosage may be used in connection
with alternate embodiments of the invention, with or without oil,
nifedipine or indomethacin. The actual dosage can depend upon the
judgment of the physician, the condition of the patient, and the
effectiveness of the therapeutic method based, for example, on the
in vitro responsiveness of relevant cultured cells or histocultured
tissue sample, or the responses observed in the appropriate animal
models.
Pharmaceutical Compositions
[0082] The instant invention also provides a pharmaceutical
composition comprising a steroid hormone such as progesterone (P4),
17-hydroxyprogesterone, 17-hydroxyprogesterone caproate,
promegestone (R5020), or pharmaceutical equivalents, analogs,
derivatives or salts thereof and a pharmaceutically acceptable
carrier. In one embodiment, the pharmaceutical composition further
comprises agents that render the steroid hormone soluble. Such
agents include but are not limited to cyclodextrins, sesame oil,
fish oil, corn oil, olive oil, coconut oil, krill oil, omega fatty
acids, mineral oil, peppermint oil, flaxseed oil, vitamin E oil,
argan oil and/or glucose solution. In another embodiment, the
pharmaceutical composition further comprises Nifedipine. In yet
another embodiment, the pharmaceutical composition further
comprises Indomethacin. In a further embodiment, the pharmaceutical
composition further comprises a steroid hormone, Nifedipine and
Indomethacin.
[0083] In one embodiment, the effective amount of the agent in the
pharmaceutical composition of about 0.05-0.1 ml/mg of steroid
hormone, 0.1-0.2 ml/mg of steroid hormone, 0.2-0.3 ml/mg of steroid
hormone, 0.3-0.4 ml/mg of steroid hormone, 0.4-0.5 ml/mg of steroid
hormone, 0.5-0.6 ml/mg of steroid hormone, 0.6-0.7 ml/mg of steroid
hormone, 0.7-0.8 ml/mg of steroid hormone, 0.8-0.9 ml/mg of steroid
hormone, 0.9-1.0 ml/mg of steroid hormone, 1.0-5.0 ml/mg of steroid
hormone, 5.0-10.0 ml/mg of steroid hormone, 10.0-15.0 ml/mg of
steroid hormone, 15.0-20.0 ml/mg of steroid hormone, 20.0-25.0
ml/mg of steroid hormone or 25.0-30.0 ml/mg of steroid hormone.
[0084] In an embodiment, the therapeutically effective amount of
the steroid hormone (such as) in the pharmaceutical composition is
about 0.5-1 mg/day, 1-5 mg/day, 5-10 mg/day, 10-15 mg/day, 15-20
mg/day, 20-25 mg/day, 25-30 mg/day, 30-35 mg/day, 35-40 mg/day,
40-45 mg/day, 45-50 mg/day, 50-55 mg/day, 55-60 mg/day, 60-65
mg/day, 65-70 mg/day, 70-75 mg/day, 75-80 mg/day, 80-85 mg/day,
85-90 mg/day, 90-95 mg/day, 95-100 mg/day, 100-200 mg/day, 200-300
mg/day, 300-500 mg/day, 500-700 mg/day, 700-1000 mg/day, 1000-2000
mg/day, 2000-3000 mg/day, 3000-4000 mg/day or 4000-5000 mg/day.
[0085] In an additional embodiment, the effective amount of
indomethacin in the pharmaceutical composition is about 0.5-1
mg/day, 1-5 mg/day, 5-10 mg/day, 10-15 mg/day, 15-20 mg/day, 20-25
mg/day, 25-30 mg/day, 30-35 mg/day, 35-40 mg/day, 40-45 mg/day,
45-50 mg/day, 50-55 mg/day, 55-60 mg/day, 60-65 mg/day, 65-70
mg/day, 70-75 mg/day, 75-80 mg/day, 80-85 mg/day, 85-90 mg/day,
90-95 mg/day or 95-100 mg/day.
[0086] In another embodiment, effective amount of nifedipine in the
pharmaceutical composition is about 0.5-1 mg/day, 1-5 mg/day, 5-10
mg/day, 10-15 mg/day, 15-20 mg/day, 20-25 mg/day, 25-30 mg/day,
30-35 mg/day, 35-40 mg/day, 40-45 mg/day, 45-50 mg/day, 50-55
mg/day, 55-60 mg/day, 60-65 mg/day, 65-70 mg/day, 70-75 mg/day,
75-80 mg/day, 80-85 mg/day, 85-90 mg/day, 90-95 mg/day or 95-100
mg/day.
[0087] In various embodiments, the present invention provides
pharmaceutical compositions including a pharmaceutically acceptable
excipient along with a therapeutically effective amount of a
steroid hormone, such as progesterone (P4), 17-hydroxyprogesterone,
17-hydroxyprogesterone caproate, promegestone (R5020), or
pharmaceutical equivalents, analogs, derivatives or salts thereof
"Pharmaceutically acceptable excipient" means an excipient that is
useful in preparing a pharmaceutical composition that is generally
safe, non-toxic, and desirable, and includes excipients that are
acceptable for veterinary use as well as for human pharmaceutical
use. Such excipients may be solid, liquid, semisolid, or, in the
case of an aerosol composition, gaseous.
[0088] In various embodiments, the pharmaceutical compositions
according to the invention may be formulated for delivery via any
route of administration. "Route of administration" may refer to any
administration pathway known in the art, including but not limited
to aerosol, nasal, oral, transmucosal, transdermal or
parenteral.
[0089] The pharmaceutical compositions according to the invention
can also contain any pharmaceutically acceptable carrier.
"Pharmaceutically acceptable carrier" as used herein refers to a
pharmaceutically acceptable material, composition, or vehicle that
is involved in carrying or transporting a compound of interest from
one tissue, organ, or portion of the body to another tissue, organ,
or portion of the body. For example, the carrier may be a liquid or
solid filler, diluent, excipient, solvent, or encapsulating
material, or a combination thereof. Each component of the carrier
must be "pharmaceutically acceptable" in that it must be compatible
with the other ingredients of the formulation. It must also be
suitable for use in contact with any tissues or organs with which
it may come in contact, meaning that it must not carry a risk of
toxicity, irritation, allergic response, immunogenicity, or any
other complication that excessively outweighs its therapeutic
benefits.
[0090] The pharmaceutical compositions according to the invention
can also be encapsulated, tableted or prepared in an emulsion or
syrup for oral administration. Pharmaceutically acceptable solid or
liquid carriers may be added to enhance or stabilize the
composition, or to facilitate preparation of the composition.
Liquid carriers include syrup, peanut oil, olive oil, glycerin,
saline, alcohols and water. Solid carriers include starch, lactose,
calcium sulfate, dihydrate, terra alba, magnesium stearate or
stearic acid, talc, pectin, acacia, agar or gelatin. The carrier
may also include a sustained release material such as glyceryl
monostearate or glyceryl distearate, alone or with a wax.
[0091] The pharmaceutical preparations are made following the
conventional techniques of pharmacy involving milling, mixing,
granulation, and compressing, when necessary, for tablet forms; or
milling, mixing and filling for hard gelatin capsule forms. When a
liquid carrier is used, the preparation will be in the form of
syrup, elixir, emulsion or an aqueous or non-aqueous suspension.
Such a liquid formulation may be administered directly per os
(p.o., by mouth) or filled into a soft gelatin capsule.
[0092] The pharmaceutical compositions according to the invention
may be delivered in a therapeutically effective amount. The precise
therapeutically effective amount is that amount of the composition
that will yield the most effective results in terms of efficacy of
treatment in a given subject. This amount will vary depending upon
a variety of factors, including but not limited to the
characteristics of the therapeutic compound (including activity,
pharmacokinetics, pharmacodynamics, and bioavailability), the
physiological condition of the subject (including age, sex, disease
type and stage, general physical condition, responsiveness to a
given dosage, and type of medication), the nature of the
pharmaceutically acceptable carrier or carriers in the formulation,
and the route of administration. One skilled in the clinical and
pharmacological arts will be able to determine a therapeutically
effective amount through routine experimentation, for instance, by
monitoring a subject's response to administration of a compound and
adjusting the dosage accordingly. For additional guidance, see
Remington: The Science and Practice of Pharmacy (Gennaro ed. 20th
edition, Williams & Wilkins PA, USA) (2000).
Kits of the Invention
[0093] The present invention is also directed to a kit to treat
and/or inhibit preterm delivery. The kit is an assemblage of
materials or components, including at least one of the inventive
compositions. Thus, in some embodiments the kit contains a
composition including progesterone or pharmaceutical equivalent,
analog, derivative, or salt thereof for topical application, as
described above.
[0094] In one embodiment, the kit is configured particularly for
the purpose of treating mammalian subjects. In another embodiment,
the kit is configured particularly for the purpose of treating
human subjects. In further embodiments, the kit is configured for
veterinary applications, treating subjects such as, but not limited
to, farm animals, domestic animals, and laboratory animals.
[0095] Instructions for use may be included in the kit.
"Instructions for use" typically include a tangible expression
describing the technique to be employed in using the components of
the kit to effect a desired outcome, such as to apply progesterone
topically. Optionally, the kit also contains other useful
components, such as, diluents, buffers, pharmaceutically acceptable
carriers, syringes, catheters, applicators, pipetting or measuring
tools, bandaging materials or other useful paraphernalia as will be
readily recognized by those of skill in the art.
[0096] The materials or components assembled in the kit can be
provided to the practitioner stored in any convenient and suitable
ways that preserve their operability and utility. For example the
components can be in dissolved, dehydrated, or lyophilized form;
they can be provided at room, refrigerated or frozen temperatures.
The components are typically contained in suitable packaging
material(s). As employed herein, the phrase "packaging material"
refers to one or more physical structures used to house the
contents of the kit, such as inventive compositions and the like.
The packaging material is constructed by well known methods,
preferably to provide a sterile, contaminant-free environment.
[0097] One skilled in the art will recognize many methods and
materials similar or equivalent to those described herein, which
could be used in the practice of the present invention. Indeed, the
present invention is in no way limited to the methods and materials
described. For purposes of the present invention, the following
terms are defined below.
Advantages of the Invention
[0098] As described herein, use of steroid hormones like
progesterone (P4), when mixed with agents that render the steroid
hormone soluble, inhibit uterine contractility and cervical
ripening, prevent preterm labor and prevent miscarriages. Agents
that render steroid hormones soluble include but are not limited to
cyclodextrins, sesame oil, fish oil, corn oil, olive oil, coconut
oil, krill oil, omega fatty acids, mineral oil, peppermint oil,
flaxseed oil, vitamin E oil, argan oil, saline solution and/or
glucose solution. Progesterone has a relatively short half life of
about 30 minutes and therefore must be administered daily.
Progesterone dissolved in saline solution (for example isotonic
saline solution) or in glucose solution may be administered by
subcutaneous injections, intravascularly (IV), vaginally, topically
or by other routes not applicable for oily compositions of
progesterone. Oily compositions of progesterone such as those
dissolved in fish oil, omega fatty acid or peppermint oil may
readily be used topically, vaginally, intramuscularly or via other
routes of administration.
[0099] In non-pregnant women, progesterone (P4) when mixed with the
aforementioned agents is useful for postmenopausal hormone
replacement therapy with and without estrogen, prevention of
amenorrhea and abnormal uterine bleeding due to hormonal imbalance
and prevention of cancer fibroids. Soluble progesterone is
particularly useful because it can be used in injections,
intravenously, subcutaneously, parenterally, topically and/or
nasally.
EXAMPLES
[0100] The following example is provided to better illustrate the
claimed invention and is not to be interpreted as limiting the
scope of the invention. To the extent that specific materials are
mentioned, it is merely for purposes of illustration and is not
intended to limit the invention. One skilled in the art may develop
equivalent means or reactants without the exercise of inventive
capacity and without departing from the scope of the invention.
[0101] As disclosed herein, the inventor evaluated cervical changes
and delivery at term during pregnancy in rats after various
progestin treatments. Pregnant rats were treated by various routes
and vehicles with progesterone (P4), 17-alpha-hydroxyprogesterone
caproate (17P), promegestone (R5020) and RU-486. Delivery time was
determined and cervical ripening assessed in vivo by collagen
light-induced fluorescence (LIF).
[0102] As further disclosed herein, the cervix is rigid in P4
injection, 17P and vaginal promegestone (R5020) groups versus
controls. Vaginal P4 had no effect. RU-486 treatment softened the
cervix during preterm delivery. Only subcutaneously P4,
promegestone (R5020) (subcutaneously and vaginal) and topical P4 in
sesame and fish oil inhibits delivery. Delivery was not changed by
subcutaneously 17P, vaginal P4, oral P4 and topical P4 in Replens.
These results demonstrate why many of the commonly used treatments
for preterm labor are not efficacious.
Example 1
Experimental Methods
Animals
[0103] Nonpregnant and timed-pregnant Sprague-Dawley rats (200-250
g) from Charles-River Laboratories (Wilmington, Mass., USA) were
delivered to animal care facilities on day 12 of gestation (day 1
being the day when a sperm plug was observed). The animals were
housed separately, with free access to food and water and
maintained on a constant 12-hour light-dark cycle. Control pregnant
rats were spontaneously delivering on day 22 and 23 of gestation.
For the measurements with the collascope the animals were
anaesthetized (interperitoneal (i.p). injection) with a combination
of xylazine (Gemini, Burns Veterinary Supply Inc, Rockville Center,
N.Y., USA) and ketamine HCl (Ketaset; Fort Dodge Laboratories Inc,
Fort Dodge, JO, USA). The animals were randomly allocated to one of
the groups and sacrificed by carbon dioxide inhalation on day 3, 5,
7 and 10 postpartum or on day 25 of pregnancy in the groups with
delayed delivery. All procedures were approved by the Animal Care
and Use Committee of the St. Joseph's Hospital and Medical Center
in Phoenix.
Treatments
[0104] Prior to any treatment LIF measurements were made in control
rats throughout pregnancy and postpartum to estimate the LIF
profile during gestation (see FIG. 1). Pregnant rats (N=6/group)
were treated (see FIG. 2), when not otherwise mentioned (see FIG.
3), from day 13 of pregnancy until delivery. Single daily
treatments were performed at 8 a.m. and twice a day treatments at 8
a.m. and 8 p.m. All single injections (4 mg P4, 10 mg 17P, 2 mg
promegestone (R5020)) were by the subcutaneous route (s.c.) in
sesame oil (0.2 ml), which was also used for the controls. Vaginal
gels were applied twice a day with a blunt ball-top needle deep
into the vagina. Crinone was used for the P4 vaginal group
(equivalent volumes of Crinone were used for 2-15 mg P4/treatment),
all data presented show the results of the highest dose (total
daily dose of 30 mg P4=1/3 of a applicator of 8% Crinone that
contains 90 mg P4). For the vaginal promegestone (R5020) group
micronized promegestone (R5020) (1 mg/treatment) was mixed into
0.18 ml of Replens. The control rats for the vaginal groups were
treated with Replens (0.18 ml/treatment). For oral P4 treatments
(15 mg, bid, vehicle sesame oil or H.sub.2O, volume 1 ml) gavage
was used. For topical P4 treatment (15 mg, bid, P4 in 1 ml sesame
oil, fish oil or in Replens) the drug was applied on the back of
animals that were shaved on day 13, 17 and 21. In some animals
(N=6) RU-486 (3 mg in 0.2 ml sesame oil) was injected
subcutaneously once on day 16 of gestation.
Reagents
[0105] Crystalline progesterone (used for oral, topical and
subcutaneous P4), RU-486, sesame oil and ethanol were purchased
from Sigma (St Louis, Mo., USA), fish oil (concentrated omega-3
fatty acids) was obtained from General Nutrition Corp. (Pittsburgh,
Pa., USA), 17-alpha-hydroxyprogesterone caproate from MP
Biomedicals (Solon, Ohio, USA), promegestone (promegestone (R5020))
from Roussel Uclaf, France. P4, 17P, promegestone (R5020) and
RU-486 were dissolved in ethanol and then mixed with sesame oil.
Crinone (micronized P4 in Replens, a bioadhesive gel) was used for
vaginal P4) and Replens were gifts from Columbia Laboratories
(Livingston, N.J., USA).
Assessment of Cervical Ripening
[0106] The amount of cervical collagen was evaluated in vivo (only
in group subcutaneously P4, vaginal P4, subcutaneously 17P, vaginal
promegestone (R5020), subcutaneously RU-486) by measurement of the
auto-fluorescent properties of cross-linked collagen with a new
prototype of an instrument, termed collascope (Reproductive
Research Technologies, Houston, Tex., USA), as used previously with
an earlier prototype.
[0107] After insertion of a small speculum into the vagina of the
anesthetized animal, the optical probe of the collascope was placed
on the surface of the exocervix. The probe, which is connected to
the main unit of the instrument by a fiberoptic cable, delivers not
only excitation light (wavelength: 339 nm) onto the cervix but also
carries the fluorescent light (mainly caused by pyridinoline
cross-links of collagen with a maximum peak at 390 nm) back to the
instrument to a CCD camera to display the full spectrum of
fluorescence and analysis of the photons emitted by the cervix. The
exposure time for excitation was 100 msec. The average of 20
measurements of the detected fluorescent intensity (photon count)
at 390 nm was used for each animal at any given time. Measurements
of cervical light-induced fluorescence (LIF) were performed on
nonpregnant animals once and in pregnant animals every other day
starting at day 13 of gestation until delivery and on postpartum
day 3 and/or postpartum day 5 (see FIGS. 2 and 3), and for some
animals also on postpartum days 7 and 10 (see FIGS. 1 and 3).
Determining the Changes in Delivery Time
[0108] Times of delivery (see also FIG. 4) of controls and various
treatment groups were determined as hours after 8 a.m. of day 22 of
gestation (FIG. 5). The expulsion of one pup was defined as
delivery.
Statistical Analyses
[0109] The cervical LIFs obtained at different times of gestation
(FIG. 1) were compared using one-way analysis of variance (ANOVA)
and multiple pairwise comparison procedures (Holm-Sidak). Student's
t-test was used to compare the LIF results of a treatment group to
its specific control group at any time in gestation and postpartum
and also to determine the differences in delivery times (FIGS. 2, 3
and 5). A two-tailed probability value of P<0.05 was considered
statistically significant.
Example 2
Effects on Preterm Delivery
LIF in Nonpregnant, Pregnant and Postpartum Rats:
[0110] Measurements of cervical light-induced-fluorescence (LIF) in
pregnant, non-treated animals show (FIG. 1) a continuously
decreasing photon count throughout pregnancy, reaching lowest
values at term, and reversal postpartum. After significant
(P<0.05) decrease from day 13 to day 15 LIF reaches a wider
plateau of non-significant (P>0.05) decreases prior to delivery.
LIF values progressively increase postpartum (P<0.05).
Effects of Injections of P4 on LIF:
[0111] LIF is significantly (P<0.05) higher in the P4 injection
group compared with vehicle controls for any day of gestation (FIG.
2A). There is no difference (P<0.05) between day 25 (delayed
delivery) in the P4 injection group (and delivery is blocked see
FIG. 4A) compared to control animals at day 21 of gestation. LIF
before treatment at day 13 shows no significant differences
(P>0.05) between the treatment and the control group and this is
similar for all treatment groups mentioned with other treatments
(FIGS. 2A-D and 3).
Effects of Vaginal P4 on LIF:
[0112] There are no significant differences (P>0.05) between the
vaginal P4 group and vehicle controls at any time in gestation
(FIG. 2B) and vaginal P4 failed to inhibit delivery (see FIG.
4B).
Effects of Injections of 17P on LIF:
[0113] LIF is significantly higher (P<0.05) in the 17P treated
group (until day 19 only) compared with vehicle controls (FIG.
2C).
Effects of Vaginal Promegestone (R5020) on LIF:
[0114] LIF is significantly higher (P<0.05) in the promegestone
(R5020) treated group (until day 19 only) compared with vehicle
controls (FIG. 2D).
Effects of a Single Injection of RU-486 on LIF:
[0115] LIF is significantly lower (P<0.05) in the RU-486 treated
group 24 and 72 hours after treatment compared with vehicle
controls (FIG. 3). LIF is higher in the RU-486 treated group 5 days
after treatment compared with vehicle controls (P<0.05).
Effects of Injections of P4 and 17P on Time of Delivery:
[0116] Injections of P4, but not 17P, completely block delivery
(FIG. 4A).
Effects of Vaginal P4 and Promegestone (R5020) on Time of
Delivery:
[0117] Vaginal promegestone (R5020), but not vaginal P4, completely
block delivery (FIG. 4B).
Effect of Various Progestins and Routes of Administration on Time
of Delivery:
[0118] FIG. 5 summarizes the results of the different treatment
groups on time of delivery and shows additional treatment groups.
Animals treated as mentioned above. Other treatment groups followed
the same design with treatments starting at day 13 of gestation
until delivery. Additional treatment groups: Injections of
promegestone (R5020) also completely block delivery. Oral P4
suspended in sesame oil or H.sub.2O had no effect on time of
delivery. However, topical P4 in sesame oil (partially) and in fish
oil (completely), but not in Replens prolongs delivery
(P<0.05).
Effects of RU-486 on Time of Delivery:
[0119] The P4 antagonist RU-486 (mifepristone) induced preterm
delivery 24-48 hours after injection (see FIG. 3; 4 of 6 animals
delivered after 24 hours and the remaining two animals delivered
within 48 hours after treatment).
Example 3
Applications of Progesterone
[0120] Various studies have raised questions about the ability of
P4 and 17P to inhibit recurrent preterm labor and whether these
progestins have effects on the cervix to prevent cervical ripening.
It is not established which of the progestins and which route of
administration is superior and there is controversy in the
findings. Existing treatment do not completely prevent preterm
birth and in many studies, women were exposed to progestins whether
they needed it or not.
[0121] As described herein, injections of P4 show (FIG. 2A) the
longest effect on delaying cervical ripening in rats of all
compounds used. This treatment inhibited the progression of
softening of the cervix in a stage of rat pregnancy, where the
physiological P4-levels are still rising..sup.33,34 Thus, higher P4
levels in midgestation of pregnancy in rat has an anti-ripening
effect on the cervix. Exogenous P4, administered by subcutaneous
injections also decelerate the consequences of the sharp withdrawal
of P4 that occurs at day 19 of rat pregnancy..sup.33,34 The
consequences of blocking the P4 receptors during pregnancy with the
P4 antagonist RU-486 results in termination of pregnancy in both
rats and humans..sup.28,35 RU-486-induced cervical ripening occurs
after 24 hours followed by preterm delivery (FIG. 3) and this
confirms effects of antiprogestins on the cervix.sup.36 and
indicates the importance of P4 in control of the cervix and
maintaining pregnancy.
[0122] Data described herein demonstrates that the intrinsic
properties of the progestins dictate their affects on the cervix
and myometrium. Thus, 17P and promegestone (R5020) delayed cervical
ripening but not at term immediately preceding delivery and
subcutaneous and vaginal promegestone (R5020) and parenteral P4
prevented delivery. Cervical ripening is only attenuated until day
19 of gestation with promegestone (R5020) and 17P in contrast to
injection of P4 which also prevents further ripening on day 21
(FIGS. 2A, 2C, 2D). This indicates that P4 accomplishes inhibitory
effects on the cervix beyond the ability of promegestone (R5020)
and 17P and demonstrates how properties of the progestins are
significant factors affecting action.
[0123] Despite any of the treatments with progestins the cervix
still manages to ripen at the end of pregnancy (FIGS. 2A-D). The
fact that prophylactic parenteral P4 can only delay, but not
completely inhibit cervical ripening indicates the involvement of
other control pathways in the ripening process. Following the
concept of the liver first-pass effect after administration of oral
drugs, de Ziegler et al. established the term "uterine first-pass
effect" to point out the minimized systemic, but optimized uterine
exposure after transvaginal treatment with sex steroids. However,
in data described herein for vaginal P4, even at a very high dose
(7.5.times. the injected dose) had no effect on cervical ripening
and on delivery
[0124] The importance of the vehicle is demonstrated by the
observations where topical P4 in fish oil completely, in sesame oil
only partially and in Replens not at all inhibits delivery (FIG.
5). This indicates that Replens may not release P4 as effectively
as oil. It could be suggested that P4 regulates parturition through
genomic actions via various proteins that are thought to be
involved in controlling myometrial contractility.
[0125] Recently, it was shown that P4, at concentrations equivalent
to those present in the placenta and uterus, inhibits spontaneous
myometrial contractility by nongenomic mechanism..sup.39 As
disclosed herein, the inventors measured delivery and cervical
ripening. Delivery at term or preterm is thought from many studies
to be due to these two main processes and involves both uterine
muscle activity and changes in the cervical connective tissue. The
inventor demonstrates that delivery is completely inhibited by some
progestin treatments (e.g. subcutaneous P4 and both subcutaneous
and vaginal promegestone (R5020)), but cervical ripening is delayed
but not entirely blocked during the final days of gestation (e.g.
subcutaneous P4) or not significantly different from controls (e.g.
vaginal promegestone (R5020)) on day 21. Therefore inhibition of
delivery is not due to an unripe cervix, but must be due to
inhibition of uterine contractions. On the other hand 17P partly
delays ripening, similar to subcutaneous P4, and is not
significantly different from controls at day 21, like promegestone
(R5020), but does not block delivery at all. RU-486, the
antiprogestin used in this study is well known to act both on the
cervix and uterus to induce delivery by stimulation of uterine
contractility and cervical ripening. Additionally the uterine
contractility increases dramatically during spontaneous delivery at
term and preterm after RU-486 treatment. 17P had no effect on
delaying term delivery supporting the conclusion that 17P is not an
effective treatment for preventing birth.
[0126] In addition use of progestins for other indications (such as
menstrual cramps, uterine and other cancers, osteoporosis,
contraception, to oppose unwanted effects of estrogens, amenorrhea
and abnormal uterine bleeding, infertility, endometriosis, etc.)
could be greatly improved by the methods described herein.
Example 4
[0127] The inventors tested whether a soluble form of progesterone
would inhibit human myometrial contractility in vitro and in vivo.
The soluble form was equally effective in inhibiting myometrial
contractility as compared to crystalline progesterone dissolved in
ethanol (FIG. 9). In addition, the inventors showed that soluble
progesterone inhibits uterine contractions (electromyographic
activity) when given to pregnant rats in labor (FIGS. 8, 15, 16,
17). Recent studies with progesterone show promise in treatment of
preterm labor but these preparations use crystalline progesterone
which is usually given vaginally. Use of soluble progesterone
offers many advantages as described above.
[0128] The inventors tested whether a soluble form of progesterone
would inhibit human myometrial contractility in vitro (FIG. 7).
FIG. 7 is a schematic drawing of the muscle bath that is used to
measure uterine contractility in vitro. The system consists of a
water jacketed chamber to maintain temperature at 37 degrees and an
inner chamber where the tissue is suspended in Krebs'-Ringer
solution and bubbled with air. The tissue is connected to a force
transducer and the signals from the transducer are amplified,
stored and analyzed by a data acquisition system.
[0129] The soluble form of progesterone (P4) was found to be
effective in inhibiting myometrial contractility (FIGS. 8, 15, 16
and 17). The inventors found that P4 inhibits myometrial
contractility (soluble and crystalline in ETOH) (FIG. 9).
Progesterone (P4) with nifedipine (Nife) (FIG. 11) or indomethacin
(IND) (FIG. 12), but not MgSO.sub.4 increased contractile
inhibition, by additive or synergistic actions. Additionally, use
of P4 with Nife or IND could also be useful for preterm labor
treatment, and higher endogenous P4 levels can increase
effectiveness of Nife or IND.
[0130] The inventors examined the direct effects of various
tocolytics on uterine contractility with and without progesterone,
as it was possible that a tocolytic used with progesterone would
produce additive, synergistic or antagonistic action on
contractility. Methods used for collecting samples and data were:
[0131] IRB approval [0132] Informed consent [0133] Women at term
not in labor (n=37 patients and 280 tissues) [0134] N=10/group
(Based on power analysis) [0135] Cesarean section [0136]
0.2.times.0.2.times.1.0 cm piece of tissue from lower uterine
segment [0137] Samples used within 24 hours of collection [0138]
Myometrial strips equilibrated with 1 gm of passive tension [0139]
Spontaneous contractile activity stabilized [0140] P4 dissolved in
ETOH added to tissue bath at 10-5 M concentration [0141] MgSO4,
indomethacin, nifedipine and pinacidil dissolved in H2O or ethanol
[0142] Solvent time-controls were run in parallel [0143]
Contractile activity registered, stored and analyzed [0144]
Statistics--One-way ANOVA used to determine statistical differences
(P<0.05=significant).
[0145] The inventors found that P4 inhibits myometrial
contractility (soluble and crystalline in ETOH). P4 with nifedipine
(Nife) or indomethacin (IND), but not MgSO.sub.4 increase
contractile inhibition, by additive or synergistic actions.
Additionally, use of P4 with Nife or IND could also be useful for
PTL treatment, and higher endogenous P4 levels can increase
effectiveness of Nife or IND. P4 decreases pinacidil inhibition of
contractility possibly by interaction of agents at K+ channels.
[0146] As described herein, progesterone applied topically in many
types of oils can inhibit delivery Soluble progesterone is usually
applied in oil preparations because of its solubility. For example,
by encapsulating progesterone in cyclodexrins progesterone becomes
soluble and can be used parenterally, and by other routes of
administration.
[0147] FIG. 15 depicts delay of delivery times in timed pregnant
rats treated with progesterone (P4) with different routes and
vehicles. Treatment was started on day 13 of gestation and
continued until delivery of pups or until delivery was blocked at
80 hours (day 25) past day 22 of gestation. 8 am on day 22 is noted
as hour 0 and delay is compared to control group treated only with
vehicle. Treatment groups included: P4 in fish oil (vaginal, nasal,
injection and topical groups), P4 in cocoa butter (rectal) and P4
in sesame oil (oral). All treatment groups received a total daily
dose of 30 mg P4/day except the nasal and subcutaneous injection
group which received 1 mg/day and 4 mg/day, respectively. Rectal,
subcutaneous injection and topical administration in fish oil had
significant delay of delivery.
[0148] As depicted in FIG. 16, pregnant timed rats were treated
topically with progesterone (P4) in different oil vehicles
beginning on day 19 of gestation. Vehicles included: Omegasorb fish
oil, coconut oil, corn oil and olive oil. Treatment was
administered twice a day (15 mg P4, bid) and continued until
delivery of pups or until complete block of delivery was observed.
8 am on day 22 is noted as hour 0 and complete block is determined
by an 80 hour delay (day 25). All groups but corn oil had
significant delay of delivery.
[0149] FIG. 17 depicts delay of delivery in time pregnant rats
treated with progesterone (P4) with different vehicles. Treatment
was started on day 20 of gestation and continued until delivery of
pups or delivery was blocked at 80 hours (day 25). 8 am on day 22
is noted as hour 0 and delay is compared to control group treated
only with vehicle. Treatment group included: DHA=Docosahexanoic
acid, EPA=Eicosapentanoic acid (both omega-3 fatty acids),
Nutraseafish oil, Nutrasea+D fish oil, GNC Triple Strength fish
oil, Neptune Krill Oil, Omegasorb GNC fish oil. All treatment
groups received a total daily dose of 30 mg P4. All treatment
groups except EPA had significant delays when compared to controls
but only Omegasorb GNC fish oil vehicle group demonstrated complete
block of delivery.
[0150] FIG. 18 depicts delay of delivery in timed pregnant rats
treated with progesterone (P4) with different vehicles. Treatment
was started on day 20 of gestation and continued until delivery of
pups or delivery was blocked at 80 hours (day 25). 8 am on day 22
is noted as hour 0 and delay is compared to control group treated
only with vehicles. Vehicles included: ETOH=Ethanol, Mineral oil,
Peppermint oil, Flaxseed oil, Vitamin E oil, Argan Oil, DMSO. All
treatment groups received a total daily dose of 30 mg P4.
[0151] As depicted in FIG. 19, animals were treated either
subcutaneously or topically with P4 in fish oil beginning on days
19, 20, 21 and 22 of gestation. Delivery time of timed pregnant
rats treated with progesterone (P4) either with subcutaneous
injection or topical application on different days of gestation.
Treatment groups consisted of P4 in fish oil administered starting
on different days of gestation and with different routes. Treatment
was administered on either days 19, 20 21 or day 22 of gestation
and continued until delivery of pups or until complete block of
delivery was observed. 8 am on day 22 is noted as Hour 0 and
complete block is determined by an 80 hour delay (day 25). All
groups had significant delay of delivery. D19 topical application
and D21 subcutaneous injection demonstrated complete block in all
animals. Delivery was blocked or significantly delayed even when
treatment was begun just prior to delivery on day 22.
[0152] As depicted in FIG. 6, topical administration of
progesterone is highly effective in increasing plasma circulation
levels of progesterone, when compared to vaginal or subcutaneous
injection, when provided at day 21. As depicted in FIG. 24, plasma
levels increase markedly after only a single treatment only day 19,
proving to be even higher than subcutaneous injection, and can
persist for 1, 2, or more days following administration.
[0153] The various methods and techniques described above provide a
number of ways to carry out the invention. Of course, it is to be
understood that not necessarily all objectives or advantages
described may be achieved in accordance with any particular
embodiment described herein. Thus, for example, those skilled in
the art will recognize that the methods can be performed in a
manner that achieves or optimizes one advantage or group of
advantages as taught herein without necessarily achieving other
objectives or advantages as may be taught or suggested herein. A
variety of advantageous and disadvantageous alternatives are
mentioned herein. It is to be understood that some preferred
embodiments specifically include one, another, or several
advantageous features, while others specifically exclude one,
another, or several disadvantageous features, while still others
specifically mitigate a present disadvantageous feature by
inclusion of one, another, or several advantageous features.
[0154] Furthermore, the skilled artisan will recognize the
applicability of various features from different embodiments.
Similarly, the various elements, features and steps discussed
above, as well as other known equivalents for each such element,
feature or step, can be mixed and matched by one of ordinary skill
in this art to perform methods in accordance with principles
described herein. Among the various elements, features, and steps
some will be specifically included and others specifically excluded
in diverse embodiments.
[0155] Although the invention has been disclosed in the context of
certain embodiments and examples, it will be understood by those
skilled in the art that the embodiments of the invention extend
beyond the specifically disclosed embodiments to other alternative
embodiments and/or uses and modifications and equivalents
thereof.
[0156] Many variations and alternative elements have been disclosed
in embodiments of the present invention. Still further variations
and alternate elements will be apparent to one of skill in the art.
Among these variations, without limitation, are sources and
composition of progestogens, including progesterone, further
including pharmaceutical compositions, preparation methods,
dosages, administration methods, and/or other diseases and
conditions and the particular use of the products created through
the teachings of the invention. Various embodiments of the
invention can specifically include or exclude any of these
variations or elements.
[0157] In some embodiments, the numbers expressing quantities of
ingredients, properties such as concentration, reaction conditions,
and so forth, used to describe and claim certain embodiments of the
invention are to be understood as being modified in some instances
by the term "about." Accordingly, in some embodiments, the
numerical parameters set forth in the written description and
attached claims are approximations that can vary depending upon the
desired properties sought to be obtained by a particular
embodiment. In some embodiments, the numerical parameters should 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
some embodiments of the invention are approximations, the numerical
values set forth in the specific examples are reported as precisely
as practicable. The numerical values presented in some embodiments
of the invention may contain certain errors necessarily resulting
from the standard deviation found in their respective testing
measurements.
[0158] In some embodiments, the terms "a" and "an" and "the" and
similar references used in the context of describing a particular
embodiment of the invention (especially in the context of certain
of the following claims) can be construed to cover both the
singular and the plural. The 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 with respect to
certain embodiments 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.
[0159] Groupings of alternative elements or embodiments of the
invention disclosed herein are not to be construed as limitations.
Each group member can be referred to and claimed individually or in
any combination with other members of the group or other elements
found herein. One or more members of a group can 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.
[0160] Preferred embodiments of this invention are described
herein, including the best mode known to the inventors for carrying
out the invention. Variations on those preferred embodiments will
become apparent to those of ordinary skill in the art upon reading
the foregoing description. It is contemplated that skilled artisans
can employ such variations as appropriate, and the invention can be
practiced otherwise than specifically described herein.
Accordingly, many embodiments of this invention include 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.
[0161] 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.
[0162] 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 can be employed
can be within the scope of the invention. Thus, by way of example,
but not of limitation, alternative configurations of the present
invention can be utilized in accordance with the teachings herein.
Accordingly, embodiments of the present invention are not limited
to that precisely as shown and described.
* * * * *