U.S. patent application number 15/355416 was filed with the patent office on 2017-03-09 for steroid analogues for neuroprotection.
The applicant listed for this patent is Emory University. Invention is credited to David B. Guthrie, Dennis C. Liotta, Mark A. Lockwood, Christopher MacNevin, Michael G. Natchus, Iqbal Sayeed, Donald G. Stein.
Application Number | 20170065615 15/355416 |
Document ID | / |
Family ID | 41016714 |
Filed Date | 2017-03-09 |
United States Patent
Application |
20170065615 |
Kind Code |
A1 |
MacNevin; Christopher ; et
al. |
March 9, 2017 |
Steroid Analogues for Neuroprotection
Abstract
Provided are steroid analogues functionalized with polar
substituents at the C3 and/or C20 positions of the steroid ring
system that exhibit improved water solubility. Also provided are
pharmaceutical compositions comprising the steroid analogues and
methods using the novel steroid analogues for the treatment and
prevention of neurodegeneration in a patient following injury to
the central nervous system.
Inventors: |
MacNevin; Christopher;
(Durham, NC) ; Stein; Donald G.; (Atlanta, GA)
; Liotta; Dennis C.; (Atlanta, GA) ; Sayeed;
Iqbal; (Atlanta, GA) ; Guthrie; David B.;
(Avondale Estates, GA) ; Lockwood; Mark A.; (Johns
Creek, GA) ; Natchus; Michael G.; (Alpharetta,
GA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Emory University |
Atlanta |
GA |
US |
|
|
Family ID: |
41016714 |
Appl. No.: |
15/355416 |
Filed: |
November 18, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12918036 |
Nov 15, 2010 |
9527881 |
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PCT/US2009/035336 |
Feb 26, 2009 |
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15355416 |
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61031567 |
Feb 26, 2008 |
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61031629 |
Feb 26, 2008 |
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61032315 |
Feb 28, 2008 |
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61148811 |
Jan 30, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07J 9/00 20130101; A61P
21/00 20180101; A61P 25/00 20180101; A61K 31/573 20130101 |
International
Class: |
A61K 31/573 20060101
A61K031/573 |
Claims
1. A method of treating or preventing neurodegeneration following
injury to the central nervous system comprising administering to a
subject in need thereof an effective amount of a compound of the
following formula: ##STR00177## or a pharmaceutically acceptable
salt thereof, wherein: R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen, hydroxyl cycloalkyl,
cycloalkenyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl,
heterocyclic, heteroaryl, amino, thiol, alkoxy, sulfide, nitro,
cyano, azide, sulfonyl, acyl, carboxyl, an ester, an amide,
carbamate, carbonate, an amino acid residue or a carbohydrate;
R.sup.3 is --OR.sup.11; R.sup.11 is --C(O)R', where R' is alkyl, or
the residue of an amino acid; and the dotted line indicates the
presence of either a single bond or a double bond, wherein valences
of a single bond are completed by hydrogens.
2. The method of claim 1, wherein neurodegeneration following
injury to the central nervous system is traumatic brain injury or
stroke.
3. The method of claim 1, wherein R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently hydrogen or alkyl.
4. The method of claim 1 wherein the compound is selected from,
##STR00178##
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a division of U.S. application Ser. No.
12/918,036 filed Aug. 17, 2010, which is the National Stage of
International Application Number PCT/US2009/035336 filed Feb. 26,
2009, which claims priority to U.S. Provisional Application No.
61/031,567 filed Feb. 26, 2008, U.S. Provisional Application No.
61/031,629 filed Feb. 26, 2008, U.S. Provisional Application No.
61/032,315 filed Feb. 28, 2008, and U.S. Provisional Application
No. 61/148,811 filed Jan. 30, 2009. The entirety of each of these
applications is hereby incorporated by reference for all
purposes.
FIELD OF THE INVENTION
[0002] The present invention is in the area of pharmaceutical
chemistry and specifically relates to certain steroid analogues,
pharmaceutical compositions and methods for neuroprotection in the
treatment and prevention of traumatic brain injury and brain injury
caused by stroke.
BACKGROUND OF THE INVENTION
[0003] Brain injuries, including traumatic brain injury (TBI) and
stroke, affect well over 2 million Americans each year and are a
significant health concern worldwide. There are currently
approximately 5.7 million stroke survivors in the US, many with
permanent disabilities, and more than 5 million Americans who have
suffered a TBI that in some cases results in the permanent need for
help in performing daily activities. Traumatic brain injuries
result from a blow or jolt to the head or a penetrating head injury
that disrupts the function of the brain, with severity ranging from
"mild," i.e., a brief change in mental status or consciousness to
"severe," i.e., an extended period of unconsciousness or amnesia
after the injury. In contrast, strokes are a result of diseases
that affect the blood vessels that supply blood to the brain. A
stroke occurs when a blood vessel that brings oxygen and nutrients
to the brain either bursts (hemorrhagic stroke) or is clogged by a
blood clot or some other mass (ischemic stroke). The majority of
strokes are ischemic, however hemorrhagic strokes typically result
in more severe injuries.
[0004] Despite several decades of effort, scientists have not yet
found a pharmacological agent that consistently improves outcomes
after stroke or TBI (see Sauerland, S. et al., Lancet 2004, 364,
1291-1292; Brain Trauma Foundation, American Association of
Neurological Surgeons, Joint Section on Neurotrauma and Critical
Care. Guidelines for the management of severe head injury. J.
Neurotrauma 1996, 13, 641-734).
[0005] After TBI or stroke, inflammation is a principle cause of
secondary damage and long-term damage. Following insults to the
central nervous system, a cascade of physiological events leads to
neuronal loss including, for example, an inflammatory immune
response and excitotoxicity resulting from disrupting the
glutamate, acetylcholine, cholinergic, GABA.sub.A, and NMDA
receptor systems. In these cases, a complex cascade of events leads
to the delivery of blood-borne leucocytes to sites of injury to
kill potential pathogens and promote tissue repair. However, the
powerful inflammatory response has the capacity to cause damage to
normal tissue, and dysregulation of the innate, or acquired immune
response is involved in different pathologies.
[0006] In addition to TBI and stroke, inflammation is being
recognized as a key component of a variety of nervous system
disorders. It has long been known that certain diseases such as
multiple sclerosis result from inflammation in the central nervous
system, but it is only in recent years that it has been suggested
that inflammation may significantly contribute to neurodegenerative
disoders such as HIV-related dementia, Alzheimer's and prion
diseases. It is now known that the resident macrophages of the
central nervous system (CNS), the microglia, when activated may
secrete molecules that cause neuronal dysfunction, or
degeneration.
[0007] There is growing experimental evidence that progesterone,
its metabolites and other gonadal steroids such as estrogen and
possibly testosterone, are effective neuroprotective agents.
Pre-clinical and clinical research demonstrates that the hormone
progesterone is a potent neurosteroid that, acutely administered,
can dramatically reduce cerebral edema, inflammation, tissue
necrosis, and programmed cell death (see Djebaili, M. et al, J.
Neurotrauma 2005, 22, 106-118; Pettus, E. H. et al., Brain Res.
2005, 1049, 112-119; Grossman, K. J. et al., Brain Res, 2004, 1008,
29-39; He, J. et al., Exp. Neurol. 2004, 189, 404-412; He, J. et
al., Restor. Neurol. Neurosci. 2004, 22, 19-31; Djebaili, M. et
al., J. Neuroscience 2004, 123, 349-359; Hoffman, S. W. et al.,
Academy of Emergency Medicine, 2001, 8, 496-497; and Wright, D. W.
et al., J. Neurotrauma. 2001, 18, 901-909).
[0008] In vivo data has demonstrated progesterone's neuroprotective
effects in injured nervous systems. For example, following a
contusion injury, progesterone reduces the severity of post injury
cerebral edema. The attenuation of edema by progesterone is
accompanied by the sparing of neurons from secondary neuronal death
and improvements in cognitive outcome (Roof et al. (1994)
Experimental Neurology 129:64-69). Furthermore, following ischemic
injury in rats, progesterone has been shown to reduce cell damage
and neurological deficit (Jiang et al. (1996) Brain Research
735:101-107). A Phase II, single-center, controlled trial involving
100 moderate to severe TBI patients showed that 3 days of
intravenous progesterone treatment reduced mortality by over 60%
and significantly improved functional outcomes at 30 days
post-injury (see Wright, D. A. et al., Ann. Emerg. Med. 2007, 49,
391).
[0009] PCT Publication WO 02/30409 to Emory University provides
methods for conferring a neuroprotective effect on a population of
cells in a subject following a traumatic injury to the central
nervous system by administration of a progestin or progestin
metabolite following a traumatic brain injury.
[0010] PCT Publication WO 06/102644 also to Emory University
provides methods for the treatment or the prevention of neuronal
damage in the CNS by tapered administration of a progestin or
progestin metabolite following a traumatic or ischemic injury to
the CNS to avoid withdrawal.
[0011] PCT Publication No. WO/2006/102596 to Emory University
provides certain methods of treating a subject with a traumatic
central nervous system injury, more particularly, a traumatic brain
injury that include a therapy comprising a constant or a two-level
dosing regime of progesterone.
[0012] Although progesterone has been shown to be neuroprotective
in traumatic brain injury, its efficacy in stroke is less well
defined. However, studies have indicated that progesterone may be
useful in treating or preventing neurodegeneration following stroke
(see Stein, D. (2005) The Case for Progesterone US Ann. N.Y. Acad.
Sci. 1052:152-169; Murphy, et al. (2002) Progesterone
Administration During Reperfusion, But Not Preischemia Alone,
Reduces Injury in Ovariectomized Rats. J. Cereb. Blood Flow &
Metab. 22:1181-1188; Murphy, et al. (2000) Progesterone Exacerbates
Striatal Stroke Injury in Progesterone-Deficient Female Animals.
Stroke 31:1173).
[0013] U.S. Pat. No. 6,245,757, now expired, to Research
Corporation Technologies, Inc. provides a method for the treatment
of ischemic damage, such as damage due to stroke or myocardial
infarction comprising administering to a mammal afflicted with
stroke an effective amount of a neuroprotective steroid in a
suitable vehicle.
[0014] In addition to being a gonadal steroid, progesterone also
belongs to a family of autocrine/paracrine hormones called
neurosteroids. Neurosteroids are steroids that accumulate in the
brain independently of endocrine sources and which can be
synthesized from sterol precursors in nervous cells. These
neurosteroids can potentiate GABA transmission, modulate the
effects of glutamate, enhance the production of myclin, and prevent
release of free radicals from activated microglia.
[0015] Various metabolites of progesterone have also been suggested
to have neuroprotective properties. For instance, the progesterone
metabolites allopregnanolone or epipregnanolone are positive
modulators of the GABA receptor, increasing the effects of GABA in
a manner that is independent of the benzodiazepines (Baulieu, E. E.
(1992) Adv. Biochem. Psychopharmacol. 47:1-16; Robel et al. (1995)
Crit. Rev. Neurobiol. 9:383-94; Lambert et al. (1995) Trends
Pharmacol. Sci. 16:295-303; Baulieu, E. E. (1997) Recent Prog.
Horm. Res. 52:1-32; Reddy et al. (1996) Psychopharmacology
128:280-92). In addition, these neurosteroids act as antagonists at
the sigma receptor, which can activate the NMDA channel complex
(Maurice et al. (1998) Neuroscience 83:413-28; Maurice et al.
(1996) J. Neurosci. Res. 46:734-43; Reddy et al. (1998) Neuroreport
9:3069-73). These neurosteroids have also been shown to reduce the
stimulation of cholinergic neurons and the subsequent release of
acetylcholine by excitability. Numerous studies have shown that the
cholinergic neurons of the basal forebrain are sensitive to injury
and that excessive release of acetylcholine can be more excitotoxic
than glutamate (Lyeth et al. (1992) J. Neurotrauma 9(2):S463-74;
Hayes et al. (1992) J. Neurotrauma 9(1):S173-87).
[0016] As discussed above, following a traumatic injury to the
central nervous system, a cascade of physiological events leads to
neuronal loss. In addition, the injury is frequently followed by
brain and/or spinal cord edema that enhances the cascade of injury
and leads to further secondary cell death and increased patient
mortality. Methods are needed for the in vivo treatment of
traumatic CNS injuries that are successful at providing subsequent
trophic support to remaining central nervous system tissue, and
thus enhancing functional repair and recovery, under the complex
physiological cascade of events which follow the initial
insult.
[0017] Although successful, the use of progesterone itself as a
therapeutic suffers from several drawbacks, including most notably
its poor solubility and its potential for generating unwanted side
effects. Therefore, there is a need for progesterone analogues that
exhibit improved solubility and bioavailability for the treatment
of ischemic, hemorrhagic or traumatic central nervous injury.
SUMMARY OF THE INVENTION
[0018] Provided are novel steroid analogues functionalized with
polar groups at the C3 and C20 positions that exhibit greater water
solubility than the parent compounds and are useful for the
prevention and treatment of nervous system damage, in particular
due to neurodegenerative reactions to injury or disease. In one
embodiment, the steroid analogues of the invention are derivatized
at the 3- and/or 20-positions of the steroid ring with polar amino
acid substituents capable of forming water soluble salts. The amino
acids may be either the naturally occurring or synthetic amino
acids in either the D, L configuration or may be a mixture of D and
L forms. In other embodiments, the steroid analogues are
derivatized at the 3- and/or 20-positions with a carbohydrate or a
substituted acyl group. The steroid analogues are optionally
substituted with non-hydrogen substituents at the 9-, 1-, 2-, 3-,
and 4-positions and may contain double bonds between C1 and C2, C4
and C5 and between C5 and C6.
[0019] In general embodiments, compounds, pharmaceutical
compositions and methods of use of a compound of Formula I are
provided:
##STR00001##
[0020] wherein X is O, N or S;
[0021] Y is O, N or S;
[0022] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0023] R.sup.4 is hydrogen or alkyl; R.sup.4 together with R.sup.3
and X forms an optionally substituted 5-6 membered heterocyclic or
heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms; or
R.sup.4 and R.sup.7 together form a double bond;
[0024] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.3 together with X and R.sup.4 forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.3 is
absent;
[0025] R.sup.7 is hydrogen or is absent, or R.sup.7 together with
R.sup.4 forms a double bond;
[0026] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.8 together with R.sup.9 and Y forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.8
absent;
[0027] R.sup.9 is hydrogen or alkyl; R.sup.9 together with Y and
R.sup.8 forms an optionally substituted 5-6 membered heterocyclic
or heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms;
or R.sup.9 and R.sup.10 together form a double bond;
[0028] R.sup.10 is hydrogen or is absent, or R.sup.10 together with
R.sup.9 forms a double bond;
[0029] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, monophosphate, diphosphate, triphosphate,
the residue of an amino acid, a carbohydrate, an optionally
substituted ester, or --C(O)R', where R' is alkyl, aryl, aralkyl,
OR.sup.1, NR.sup.1R.sup.2 or the residue of an amino acid;
[0030] R.sup.12 is hydrogen or alkyl; and the dotted line indicates
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, with certain
provisos.
[0031] In more specific embodiments, a residue of an amino acid is
connected to the steroid ring system at the carboxyl group of the
amino acid. In other embodiments, a residue of an amino acid is
connected to the steroid at the amino acid side chain. For example,
amino acids that contain side chains with functional groups that
are capable of forming a bond with a hydroxy or a ketone group may
be bonded to the progestin ring by such a group. In other
embodiments, the reactive groups on the amino acid may displace
leaving groups formed on the progestin moiety to form a covalent
bond. Non-limiting examples of amino acids with reactive groups in
the side chain include lysine, cysteine, serine, tyrosine, aspartic
acid, arginine and the like.
[0032] In another embodiment, the enantiomers of the compounds of
Formula I are provided. In this embodiment, the stereochemical
configuration of each asymmetric carbon is opposite that of the
natural steroids and analogues of the natural steroids.
[0033] In specific embodiments of the invention, methods of
treating or preventing damage resulting from nervous system injury
or disease are provided, such as from traumatic brain injury or
stroke, comprising administering a compound of Formula I or a
pharmaceutically acceptable salt, ester or prodrug thereof in a
pharmaceutically acceptable carrier to a patient in need thereof.
In some embodiments, the damage is due to inflammation initiated by
a TBI. In other embodiments, methods of treating or preventing
neurodegeneration resulting from ischemic CNS injuries, in
particular from ischemic stroke are provided comprising
administering a compound of the invention to a patient in need
thereof. In yet other embodiments, methods of treating or
preventing neurodegeneration resulting from hemorrhagic CNS
injuries, in particular from hemorrhagic stroke are provided
comprising administering a compound of the invention to a patient
in need thereof. The methods can alleviate the initial damage to
the CNS. Therefore, in some embodiments, the compounds are
administered to a patient at risk of a CNS injury, in particular to
a patient at risk of a stroke. The compounds are also effective at
reducing or preventing secondary injuries. Therefore, in other
embodiments, the compounds are administered to a patient who has
suffered a CNS injury within a window of opportunity after the
initial insult. The initial insult can be either a TBI or a stroke,
whether that be an ischemic or hemorrhagic stroke. In other
embodiments, the neural damage occurs from a disease or disorder
such as multiple sclerosis or Amyotrophic Lateral Sclerosis (ALS).
Pharmaceutical compositions, including in combination with
additional neuroprotective agents, are also provided.
BRIEF DESCRIPTION OF THE FIGURES
[0034] FIG. 1 shows cerebral edema assay data for selected steroid
analogues.
[0035] FIG. 2 shows the solubility of selected steroid analogues in
phosphate buffered saline.
[0036] FIG. 3 shows pharmacokinetic data of steroid analogues P1-31
and P1-33 after dosing at 10 mg/kg.
[0037] FIG. 4 shows pharmacokinetic data of steroid analogue P1-131
after dosing at 10 mg/kg.
[0038] FIG. 5 shows pharmacokinetic data of steroid analogue P1-186
after dosing at 10 mg/kg.
DETAILED DESCRIPTION OF THE INVENTION
[0039] Progesterone is lipid-soluble and essentially water
insoluble. Therefore, the intravenous administration of
progesterone is difficult in a clinical setting and the dose is
limited by the low aqueous solubility of the compound. The present
invention provides novel steroid analogues that comprise polar
groups and exhibit increased aqueous solubility. Also provided are
pharmaceutically acceptable salts, esters and prodrugs of the
steroid analogues. The compounds, salts, esters and prodrugs
provided are useful for the treatment or prevention of central
nervous system injury, particularly for the treatment or prevention
of damage arising from a traumatic brain injury and the treatment
or prevention of injuries occurring from stroke.
[0040] The term "steroid analogues" as used herein is intended to
encompass analogues of progesterone, analogues of progesterone
metabolites or derivatives and other non-progestin steroid
compounds. The steroid analogues of the invention exhibit increased
solubility in aqueous solvents and are capable of forming
pharmaceutically acceptable salts that further increase their
aqueous solubility. Also provided are pharmaceutical compositions
comprising the inventive steroid analogues, pharmaceutically
acceptable salts, esters or prodrugs thereof, and methods for the
treatment or prevention of CNS injuries, including traumatic brain
injury and stroke. In particular, the present invention relates to
steroid analogues that comprise amino acid residues, carbohydrates
or other suitable polar groups at the 3- and/or 20-positions of the
steroid ring system. The improved water solubility of the steroid
analogues of the invention can facilitate the administration of the
compounds, in particular intravenous administration, which provides
the fastest possible exposure of the active agent to the brain or
other CNS sites where it is needed, increasing the efficacy of the
drug. In addition, the inventive steroid analogs can minimize
undesired side effects that are typically accompany acute or
prolonged treatment with progesterone, such as sleepiness, reduced
arousal and increased blood clotting.
I. Compounds of the Invention
[0041] The compounds of the present invention are steroid analogues
that are modified to comprise polar groups and exhibit a greater
aqueous solubility than the naturally occurring parent compounds.
In one embodiment, analogues of steroid compounds are provided that
are modified at the 3- and/or 20-position of the steroid ring
system to incorporate polar groups. The ring numbering shown below
for the structure of progesterone (PROG) is maintained throughout
this document to avoid ambiguity.
##STR00002##
[0042] Substituents on the steroid analogues that lie below the
plane of the paper as drawn are termed in the ".alpha." or "alpha"
configuration. Substituents that lie above the plane of the paper
are termed in the ".beta." or "beta" configuration. For example the
two methyl groups shown in the progesterone structure below are in
the beta configuration.
[0043] In one embodiment of the invention are provided steroid
analogues, such as progesterone, pregnenolone and the like,
comprising an amino acid residue, a carbohydrate or other polar
group bonded to the 3-position of the steroid ring system. In
another embodiment of the invention, steroid analogues that
comprise an amino acid residue, a carbohydrate or other polar group
bonded to the 20-position of the ring system are provided. In still
another embodiment, steroid analogues comprising amino acid
residues and/or carbohydrates or other polar groups at the 3- and
at the 20-positions of the ring system are provided. The inventive
steroid analogues have greater aqueous solubility than the parent
compounds and are thus advantageous for administration, in
particular in situations in which rapid availability and effective
dosing of the compounds are critical. In some embodiments, the
steroid analogues comprise a basic nitrogen group that enables the
formation of pharmaceutically acceptable salts and prodrugs. The
inventive steroid analogues are useful for the treatment or
prevention of central nervous system injury, particularly traumatic
brain injury and stroke.
[0044] In one embodiment, the invention provides a steroid analogue
of Formula I:
##STR00003##
[0045] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0046] wherein X is O, N or S;
[0047] Y is O, N or S;
[0048] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0049] R.sup.4 is hydrogen or alkyl; R.sup.4 together with R.sup.3
and X forms an optionally substituted 5-6 membered heterocyclic or
heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms; or
R.sup.4 and R.sup.7 together form a double bond;
[0050] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.3 together with X and R.sup.4 forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.3 is
absent;
[0051] R.sup.7 is hydrogen or is absent, or R.sup.7 together with
R.sup.4 forms a double bond;
[0052] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.8 together with R.sup.9 and Y forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.8
absent;
[0053] R.sup.9 is hydrogen, alkyl; R.sup.9 together with Y and
R.sup.8 forms an optionally substituted 5-6 membered heterocyclic
or heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms,
or R.sup.9 and R.sup.10 together form a double bond;
[0054] R.sup.10 is hydrogen or is absent, or R.sup.10 together with
R.sup.9 forms a double bond;
[0055] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, monophosphate, diphosphate, triphosphate,
the residue of an amino acid, a carbohydrate, an optionally
substituted ester, or --C(O)R', where R' is OR.sup.1,
NR.sup.1R.sup.2, alkyl, aryl, aralkyl, or the residue of an amino
acid;
[0056] R.sup.12 is hydrogen or alkyl; and the dotted line indicates
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
at least one of XR.sup.3R.sup.7 or YR.sup.8R.sup.10 is not .dbd.O
or OH; and that if the dotted line between C4 and C5 or between C5
and C6 represents a double bond then the other dotted line between
C4 and C5 or between C5 and C6 represents a single bond; and with
the proviso that neither XR.sup.3R.sup.7 nor YR.sup.8R.sup.10
represent an ester of aspartic acid, glutamic acid, gamma amino
butyric acid or .alpha.-2-(hydroxyethylamino)-propionic acid; and
with the proviso that when Y is N, R.sup.8 does not represent
aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0057] In one embodiment of Formula I, R.sup.3 is hydrogen,
optionally substituted acyl, a residue of an amino acid, a
carbohydrate, --OR.sup.11, --NR.sup.11R.sup.12 or R.sup.3 is
absent; R.sup.4 is hydrogen or alkyl, or R.sup.4 and R.sup.7
together form a double bond; R.sup.8 is hydrogen, optionally
substituted acyl, a residue of an amino acid, a carbohydrate,
--OR.sup.11, --NR.sup.11R.sup.12, or R.sup.8 is absent; R.sup.9 is
hydrogen or alkyl, or R.sup.9 and R.sup.10 together form a double
bond; and R.sup.11 is a residue of an amino acid, a carbohydrate,
or an optionally substituted ester.
[0058] In some embodiments, compounds of Formula I are provided
wherein: R.sup.3 is hydrogen, optionally substituted acyl, a
residue of an amino acid, a carbohydrate, --OR.sup.11,
--NR.sup.11R.sup.12 or R.sup.3 is absent; R.sup.8 is hydrogen,
optionally substituted acyl, a residue of an amino acid, a
carbohydrate, --OR.sup.11, --NR.sup.11R.sup.12 or R.sup.8 absent;
and R.sup.11 is the residue of an amino acid, a carbohydrate or an
optionally substituted ester.
[0059] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0060] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0061] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0062] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0063] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0064] In still another embodiment, R.sup.4 and R.sup.9 are
independently hydrogen or methyl.
[0065] In some embodiments, X and Y are O. In other embodiments, X
is O and Y is N or X is N and Y is O. In other embodiments, both X
and Y are N. In certain embodiments in which Y is O, R.sup.9 and
R.sup.10 come together to form a double bond.
[0066] In certain embodiments, one of R.sup.3 and R.sup.8 is a
residue of an amino acid. In particular embodiments, the amino acid
is a naturally occurring amino acid. In certain embodiments,
R.sup.3 is a residue of an amino acid. In certain other
embodiments, R.sup.8 is a residue of an amino acid. In yet further
embodiments, both R.sup.3 and R.sup.8 are residues of an amino
acid.
[0067] In other embodiments, R.sup.3 is a carbohydrate. In another
embodiment, R.sup.8 is a carbohydrate.
[0068] In one embodiment of Formula I, X is O, R.sup.3 is the
residue of an amino acid, and R.sup.7 is absent.
[0069] In another embodiment of Formula I, Y is O, R.sup.8 is the
residue of an amino acid, and R.sup.10 is absent;
[0070] In another embodiment of Formula I, X is N; R.sup.7 together
with R.sup.4 form a double bond; R.sup.3 is OR.sup.11 or
NR.sup.11R.sup.12; and R.sup.11 is the residue of an amino acid or
a carbohydrate.
[0071] In another embodiment of Formula I, Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is OR.sup.11 or
NR.sup.11R.sup.12; and R.sup.11 is the residue of an amino acid or
a carbohydrate.
[0072] In another embodiment of Formula I, X is O; R.sup.3 is the
residue of a naturally occurring amino acid or a carbohydrate;
R.sup.7 is absent; Y is O; R.sup.8 is absent; and R.sup.9 and
R.sup.10 together form a double bond.
[0073] In still another embodiment, Y is O; R.sup.8 is the residue
of a naturally occurring amino acid or a carbohydrate; R.sup.10 is
absent; X is O; R.sup.7 is absent; and R.sup.3 and R.sup.4 together
form a double bond.
[0074] In another embodiment of Formula I, X is O; R.sup.3 is the
residue of an amino acid; R.sup.7 is absent; Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is OR.sup.11 or
NR.sup.11R.sup.12; and R.sup.11 is the residue of an amino acid or
a carbohydrate.
[0075] In yet another embodiment of Formula I, X is N; R.sup.7
together with R.sup.4 form a double bond; R.sup.3 is OR.sup.11 or
NR.sup.11R.sup.12; R.sup.11 is the residue of an amino acid or a
carbohydrate; Y is O; R.sup.8 is the residue of an amino acid; and
R.sup.10 is absent.
[0076] In another embodiment of Formula I, X is O; R.sup.3 is the
residue of an amino acid or a carbohydrate; R.sup.7 is absent; Y is
O, R.sup.8 is the residue of an amino acid or a carbohydrate, and
R.sup.10 is absent.
[0077] In yet another embodiment of Formula I, X is N; R.sup.7
together with R.sup.4 form a double bond; R.sup.3 is OR.sup.11 or
NR.sup.11R.sup.12; Y is N; R.sup.10 together with R.sup.9 form a
double bond; R.sup.8 is OR.sup.11 or NR.sup.11R.sup.12; and
R.sup.11 is the residue of an amino acid or a carbohydrate.
[0078] In another embodiment, X is N, R.sup.7 together with R.sup.4
form a double bond, and R.sup.3 is --C(O)R'.
[0079] In another embodiment, Y is N, R.sup.9 and R.sup.10 together
form a double bond, and R8 is --C(O)R'.
[0080] In another embodiment, X is O and R.sup.3 is monophosphate,
diphosphate, triphosphate, In another embodiment, Y is O and
R.sup.8 is monophosphate, diphosphate, triphosphate,
[0081] In yet another embodiment of Formula I, X is O; R.sup.3 is
the residue of an amino acid; R.sup.7 is absent; R.sup.1, R.sup.2,
R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen, alkyl,
halogen or hydroxyl.
[0082] In yet another embodiment of Formula I, X is O; R.sup.3 is
the residue of an amino acid; R.sup.7 is absent; R.sup.1, R.sup.2,
R.sup.4, R.sup.5 and R.sup.6 are hydrogen.
[0083] In another embodiment of Formula I, Y is O; R.sup.8 is the
residue of an amino acid; R.sup.10 is absent; R.sup.1, R.sup.2,
R.sup.4, R.sup.5 and R.sup.6 are independently hydrogen, alkyl,
halogen or hydroxyl.
[0084] In still another embodiment of Formula I, Y is O; R.sup.8 is
the residue of an amino acid; R.sup.10 is absent; R.sup.1, R.sup.2,
R.sup.4, R.sup.5 and R.sup.6 are hydrogen.
[0085] In one embodiment of Formula I, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0086] In another embodiment of Formula I, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0087] In another embodiment of Formula I, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0088] In still another embodiment, the dotted line between C1 and
C2 represents a single bond. In another embodiment, the dotted line
between C1 and C2 represents a double bond.
[0089] In certain embodiments of Formula I, a residue of an amino
acid is connected to the steroid ring system at the carboxyl group
of the amino acid. In other embodiments, a residue of an amino acid
is connected to the steroid at the amino acid side chain. For
example, amino acids that contain side chains with functional
groups that are capable of forming a bond with a hydroxy or a
ketone group may be boded to the steroid ring by such a group. In
other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid, arginine and the like.
[0090] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0091] In one embodiment, a compound of Formula I with the
stereochemical configuration of Formula Ia is provided:
##STR00004##
[0092] wherein the definition of variables X, Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 and
R.sup.10 is as defined in Formula I above.
[0093] In another embodiment, a compound of Formula I with the
stereochemical configuration of Formula Ib is provided:
##STR00005##
[0094] wherein the definition of variables X, Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9 and
R.sup.10 is as defined in Formula I above.
[0095] In another embodiment, a progesterone or steroid analogue
Formula II is provided:
##STR00006##
[0096] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0097] wherein Y is O, N or S;
[0098] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0099] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid, monophosphate, diphosphate, triphosphate, a
carbohydrate; --C(O)R', where R' is alkyl, aryl, aralkyl, OR.sup.1,
NR.sup.1R.sup.2 or the residue of an amino acid; or R.sup.3
together with the oxygen and R.sup.4 forms an optionally
substituted 5-6 membered heterocyclic or heteroaryl ring containing
1-4 nitrogen, oxygen or sulfur atoms;
[0100] R.sup.4 is hydrogen or alkyl, or R.sup.4 together with
R.sup.3 and the oxygen atom forms an optionally substituted 5-6
membered heterocyclic or heteroaryl ring containing 1-4 nitrogen,
oxygen or sulfur atoms;
[0101] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.8 together with R.sup.9 and Y forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.8 is
absent;
[0102] R.sup.9 is hydrogen or alkyl; R.sup.9 together with Y and
R.sup.8 forms an optionally substituted 5-6 membered heterocyclic
or heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms;
or R.sup.9 and R.sup.10 together form a double bond;
[0103] R.sup.10 is hydrogen or absent, or R.sup.10 together with
R.sup.9 form a double bond;
[0104] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, monophosphate, diphosphate, triphosphate,
the residue of an amino acid, a carbohydrate, optionally
substituted acyl, or --C(O)R', where R' is alkyl, aryl, aralkyl,
OR.sup.1, NR.sup.1R.sup.2, or the residue of an amino acid;
[0105] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond; and with the
proviso that neither R.sup.3 nor YR.sup.8R.sup.10 represent an
ester of aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid; and with the proviso
that when Y is N, R.sup.8 does not represent aspartic acid,
glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0106] In one embodiment of Formula II, R.sup.3 is hydrogen,
optionally substituted acyl, a residue of an amino acid or a
carbohydrate; R.sup.8 is hydrogen, optionally substituted acyl, a
residue of an amino acid, a carbohydrate, --OR.sup.11,
--NR.sup.11R.sup.12 or R.sup.8 is absent; and R.sup.11 is the
residue of an amino acid, a carbohydrate or optionally substituted
acyl.
[0107] In some embodiments, Y is O. In other embodiments, Y is N.
In certain embodiments in which Y is O, R.sup.9 and R.sup.10 come
together to form a double bond. In certain embodiments, one of
R.sup.3 and R.sup.8 is a residue of an amino acid. In particular
embodiments, the amino acid is a naturally occurring amino acid. In
certain embodiments, R.sup.3 is a residue of an amino acid. In
certain other embodiments, R.sup.8 is a residue of an amino acid.
In yet further embodiments, both R.sup.3 and R.sup.8 are residues
of an amino acid.
[0108] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0109] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0110] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0111] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0112] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0113] In still another embodiment, R.sup.4 and R.sup.9 are
independently hydrogen or methyl.
[0114] In one embodiment of Formula II, Y is O; R.sup.8 is the
residue of an amino acid; and R.sup.10 is absent.
[0115] In another embodiment of Formula II, Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is OR.sup.11; and
R.sup.11 is the residue of an amino acid or a carbohydrate.
[0116] In another embodiment of Formula II, Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of an amino acid or a
carbohydrate; and R.sup.12 is hydrogen.
[0117] In another embodiment of Formula II, R.sup.3 is the residue
of a naturally occurring amino acid or a carbohydrate; R.sup.4 is
hydrogen; Y is O; R.sup.10 together with R.sup.9 form a double
bond; and R.sup.8 is absent.
[0118] In another embodiment of Formula II, R.sup.3 is a
carbohydrate; R.sup.4 is hydrogen; Y is O; R.sup.10 together with
R.sup.9 form a double bond; and R.sup.8 is absent.
[0119] In another embodiment of Formula II, R.sup.3 is the residue
of a naturally occurring amino acid or a carbohydrate; R.sup.4 is
hydrogen; Y is O; R.sup.8 and R.sup.9 are hydrogen; and R.sup.10 is
absent.
[0120] In another embodiment of Formula II, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula II, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl.
[0121] In another embodiment, R.sup.1 and R.sup.2 are hydroxyl. In
still another embodiment, R.sup.1 and R.sup.2 are independently
hydroxyl or halogen. In another embodiment of Formula II, R.sup.1
is alkyl; and R.sup.2 and R.sup.5 are hydrogen. In another
embodiment, of Formula II, R.sup.2 is alkyl; and R.sup.1 and
R.sup.5 are hydrogen. In still another embodiment, of Formula II,
R.sup.5 is alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0122] In another embodiment of Formula II, R.sup.3 is the residue
of a naturally occurring amino acid; R.sup.4 is hydrogen; Y is O;
R.sup.10 together with R.sup.9 form a double bond; R.sup.8 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0123] In another embodiment of Formula II, R.sup.3 is the residue
of a naturally occurring amino acid; R.sup.4 is alkyl; Y is O;
R.sup.10 together with R.sup.9 form a double bond; R.sup.8 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0124] In one embodiment of Formula II, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0125] In another embodiment of Formula II, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0126] In another embodiment of Formula II, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0127] In still another embodiment of Formula II, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0128] In another embodiment of Formula II, OR.sup.3 is in the
alpha configuration. In still another embodiment, OR.sup.3 is in
the beta configuration.
[0129] In one embodiment of Formula II, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0130] In one embodiment of Formula II, R.sup.3 represents a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.3 is a residue of L-valine. In other
embodiments, R.sup.3 represents an amino acid residue with the
D-configuration or R.sup.3 represents a non-natural amino acid. In
other embodiments, R.sup.3 represents the residue of a .beta.
.gamma. or .delta. amino acid.
[0131] In one preferred embodiment of Formula II, R.sup.3
represents an ester of an amino acid. In another embodiment,
R.sup.3 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid. In certain embodiments of Formula II, a residue of
an amino acid is connected to the steroid ring system at the
carboxyl group of the amino acid. In other embodiments, a residue
of an amino acid is connected to the steroid at the amino acid side
chain. For example, amino acids that contain side chains with
functional groups that are capable of forming a bond with a hydroxy
or a ketone group may be boded to the steroid ring by such a group.
In other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid and arginine.
[0132] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0133] In one embodiment, a compound of Formula II with the
stereochemical configuration of Formula IIa is provided:
##STR00007##
[0134] wherein the definition of variables Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.8, R.sup.9 and R.sup.10
are as defined in Formula II above.
[0135] In still another embodiment, a compound of Formula II with
the stereochemical configuration of Formula IIb is provided:
##STR00008##
[0136] wherein the definition of variables Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.8, R.sup.9 and R.sup.10
are as defined in Formula II above.
[0137] In another embodiment, a progesterone analogue of Formula
III is provided:
##STR00009##
[0138] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0139] wherein X is O, N or S;
[0140] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0141] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate, --OR.sup.11, --NR.sup.11R.sup.12, --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid; R.sup.3 together with X and R.sup.4 forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.3 is
absent;
[0142] R.sup.4 is hydrogen or alkyl; R.sup.4 together with R.sup.3
and X forms an optionally substituted 5-6 membered heterocyclic or
heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms; or
R.sup.4 together with R.sup.7 form a double bond;
[0143] R.sup.7 is hydrogen or is absent, or R.sup.7 together with
R.sup.4 forms a double bond;
[0144] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, monophosphate, diphosphate, triphosphate, a
carbohydrate; --C(O)R', where R' is alkyl, aryl, aralkyl, OR.sup.1,
NR.sup.1R.sup.2 or the residue of an amino acid; or R.sup.8
together with R.sup.9 and the oxygen atom forms an optionally
substituted 5-6 membered heterocyclic or heteroaryl ring containing
1-4 nitrogen, oxygen or sulfur atoms;
[0145] R.sup.9 is hydrogen, alkyl, or R.sup.9 together with the
oxygen atom and R.sup.8 forms an optionally substituted 5-6
membered heterocyclic or heteroaryl ring containing 1-4 nitrogen,
oxygen or sulfur atoms;
[0146] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, a
carbohydrate, optionally substituted acyl, or --C(O)R', where R' is
alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2, or the residue of
an amino acid;
[0147] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond; and with the
proviso that neither XR.sup.3R.sup.7 nor R.sup.8 represent an ester
of aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0148] In one embodiment of Formula III, R.sup.3 is hydrogen,
optionally substituted acyl, a residue of an amino acid, a
carbohydrate, --OR.sup.11, --NR.sup.11R.sup.12 or R.sup.3 is
absent; R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, or a carbohydrate; R.sup.11 is the residue of an
amino acid, a carbohydrate or optionally substituted acyl.
[0149] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0150] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0151] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0152] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0153] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0154] In still another embodiment, R.sup.4 and R.sup.9 are
independently hydrogen or methyl.
[0155] In some embodiments, X is O. In other embodiments, X is N.
In certain embodiments in which X is O, R.sup.3 and R.sup.4 come
together to form a double bond. In certain embodiments, one of
R.sup.3 and R.sup.8 is a residue of an amino acid. In particular
embodiments, the amino acid is a naturally occurring amino acid. In
certain embodiments, R.sup.3 is a residue of an amino acid. In
certain other embodiments, R.sup.8 is a residue of an amino acid.
In yet further embodiments, both R.sup.3 and R.sup.8 are residues
of an amino acid.
[0156] In one embodiment, R.sup.3 is a carbohydrate. In another
embodiment, R.sup.8 is a carbohydrate.
[0157] In one embodiment of Formula III, X is O; R.sup.3 is the
residue of an amino acid or a carbohydrate; and R.sup.7 is
absent.
[0158] In another embodiment of Formula III, X is N; R.sup.4
together with R.sup.7 form a double bond; R.sup.3 is OR.sup.11; and
R.sup.11 is the residue of an amino acid or a carbohydrate.
[0159] In another embodiment of Formula III, X is N; R.sup.4
together with R.sup.7 form a double bond; R.sup.3 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of an amino acid or a
carbohydrate; and R.sup.12 is hydrogen.
[0160] In another embodiment of Formula III, R.sup.8 is the residue
of a naturally occurring amino acid; R.sup.9 is hydrogen; X is O;
R.sup.4 together with R.sup.7 form a double bond; and R.sup.3 is
absent.
[0161] In another embodiment of Formula III, R.sup.8 is a
carbohydrate; R.sup.9 is hydrogen; X is O; R.sup.4 together with
R.sup.7 form a double bond; and R.sup.3 is absent.
[0162] In another embodiment of Formula III, R.sup.8 is the residue
of a naturally occurring amino acid or a carbohydrate; R.sup.9 is
hydrogen; X is O; R.sup.3 and R.sup.4 are hydrogen; and R.sup.7 is
absent.
[0163] In another embodiment of Formula III, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula III, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula III, R.sup.1 is alkyl;
and R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of
Formula III, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are
hydrogen. In still another embodiment, of Formula III, R.sup.5 is
alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0164] In another embodiment of Formula III, R.sup.8 is the residue
of a naturally occurring amino acid; R.sup.9 is hydrogen; X is O;
R.sup.4 together with R.sup.7 form a double bond; R.sup.3 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0165] In another embodiment of Formula III, R.sup.8 is the residue
of a naturally occurring amino acid; R.sup.9 is alkyl; X is O;
R.sup.4 together with R.sup.7 form a double bond; R.sup.3 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0166] In one embodiment of Formula III, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0167] In another embodiment of Formula III, the dotted line
between C4 and C5 represents a single bond and the dotted line
between C5 and C6 represents a double bond.
[0168] In another embodiment of Formula III, the dotted line
between C4 and C5 represents a double bond and the dotted line
between C5 and C6 represents a single bond.
[0169] In still another embodiment of Formula III, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0170] In another embodiment of Formula III, --XR.sup.3R.sup.7 is
in the alpha configuration. In still another embodiment,
--XR.sup.3R.sup.7 is in the beta configuration.
[0171] In one embodiment of Formula III, the dotted lines between
C4 and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0172] In one embodiment of Formula III, R.sup.8 represents a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.8 is a residue of L-valine. In another
embodiment, R.sup.8 represents an amino acid residue with the
D-configuration. In another embodiment, R.sup.8 represents a
non-natural amino acid. In other embodiments, R.sup.8 represents
the residue of a .beta. .gamma. or .delta. amino acid.
[0173] In one preferred embodiment of Formula III, R.sup.8
represents an ester of an amino acid. In another embodiment,
R.sup.8 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid.
[0174] In one preferred embodiment of Formula III, R.sup.3
represents an ester of an amino acid. In another embodiment,
R.sup.3 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid. In certain embodiments of Formula III, a residue of
an amino acid is connected to the steroid ring system at the
carboxyl group of the amino acid. In other embodiments, a residue
of an amino acid is connected to the steroid at the amino acid side
chain. For example, amino acids that contain side chains with
functional groups that are capable of forming a bond with a hydroxy
or a ketone group may be boded to the steroid ring by such a group.
In other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid, arginine and the like.
[0175] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0176] In one embodiment, a compound of Formula III with the
stereochemical configuration of Formula IIIa is provided:
##STR00010##
[0177] wherein the definition of variables X, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, and R.sup.9
are as defined in Formula III above.
[0178] In yet another embodiment, a compound of Formula III with
the stereochemical configuration of Formula IIIb is provided:
##STR00011##
[0179] wherein the definition of variables X, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, and R.sup.9
are as defined in Formula III above.
[0180] In another embodiment a compound of Formula IV is
provided:
##STR00012##
[0181] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0182] wherein Y is O, N or S;
[0183] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0184] R.sup.3 is an --OR.sup.11, --NR.sup.11R.sup.12,
monophosphate, diphosphate, triphosphate, a carbohydrate, or
--C(O)R', where R' is alkyl, aryl, aralkyl, OR.sup.1,
NR.sup.1R.sup.2 or the residue of an amino acid;
[0185] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, --OR.sup.11, --NR.sup.11R.sup.12,
monophosphate, diphosphate, triphosphate, --C(O)R', where R' is
alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue of
an amino acid; or R.sup.8 together with R.sup.9 and Y forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.8 is
absent;
[0186] R.sup.9 is hydrogen or alkyl; R.sup.9 together with Y and
R.sup.8 forms an optionally substituted 5-6 membered heterocyclic
or heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms,
or R.sup.9 and R.sup.10 together form a double bond;
[0187] R.sup.10 is hydrogen or absent, or R.sup.10 together with
R.sup.9 form a double bond;
[0188] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, a
carbohydrate monophosphate, diphosphate, triphosphate; --C(O)R',
where R' is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the
residue of an amino acid; or an optionally substituted ester;
[0189] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond; and with the
proviso that YR.sup.8R.sup.10 does not represent an ester of
aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid; and with the proviso
that when Y is N, R.sup.8 does not represent aspartic acid,
glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0190] In one embodiment of Formula IV, R.sup.3 is an --OR.sup.11,
--NR.sup.11R.sup.12, or a carbohydrate; R.sup.8 is hydrogen,
optionally substituted acyl, a residue of an amino acid, a
carbohydrate, --OR.sup.11, --NR.sup.11R.sup.12, or R.sup.8 is
absent; R.sup.11 is the residue of an amino acid, a carbohydrate or
an optionally substituted ester;
[0191] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0192] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0193] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0194] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0195] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0196] In still another embodiment, R.sup.9 is hydrogen or
methyl.
[0197] In some embodiments, Y is O. In other embodiments, Y is N.
In certain embodiments in which Y is O, and R.sup.8 and R.sup.10
come together to form a double bond. In certain other embodiments,
one of R.sup.3 and R.sup.8 is a residue of an amino acid. In
particular embodiments, the amino acid is a naturally occurring
amino acid. In certain embodiments, R.sup.3 is a residue of an
amino acid. In certain other embodiments, R.sup.8 is a residue of
an amino acid. In yet further embodiments, both R.sup.3 and R.sup.8
are residues of an amino acid.
[0198] In other embodiments, R.sup.3 is a carbohydrate. In another
embodiment, R.sup.8 is a carbohydrate.
[0199] In one embodiment of Formula IV, Y is O; R.sup.8 is the
residue of an amino acid; and R.sup.10 is absent.
[0200] In another embodiment of Formula IV, Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is OR.sup.11; and
R.sup.11 is the residue of an amino acid or a carbohydrate.
[0201] In another embodiment of Formula IV, Y is N; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of an amino acid or a
carbohydrate; and R.sup.12 is hydrogen.
[0202] In another embodiment of Formula IV, R.sup.3 is --OR.sup.11
and R.sup.11 is the residue of a naturally occurring amino acid or
a carbohydrate; Y is O; R.sup.10 together with R.sup.9 form a
double bond; and R.sup.8 is absent.
[0203] In another embodiment of Formula IV, R.sup.3 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; Y is O; R.sup.10
together with R.sup.9 form a double bond; and R.sup.8 is
absent.
[0204] In another embodiment of Formula IV, R.sup.3 is a
carbohydrate; Y is O; R.sup.10 together with R.sup.9 form a double
bond; and R.sup.8 is absent.
[0205] In another embodiment of Formula IV, R.sup.3 is --OR.sup.11
and R.sup.11 is the residue of a naturally occurring amino acid or
a carbohydrate; Y is O; R.sup.8 and R.sup.9 are hydrogen; and
R.sup.10 is absent.
[0206] In another embodiment of Formula IV, R.sup.3 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; Y is O; R.sup.8 and
R.sup.9 are hydrogen; and R.sup.10 is absent.
[0207] In another embodiment of Formula IV, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula IV, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula IV, R.sup.1 is alkyl; and
R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of Formula
IV, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are hydrogen. In
still another embodiment, of Formula IV, R.sup.5 is alkyl; and
R.sup.1 and R.sup.2 are hydrogen.
[0208] In another embodiment of Formula IV, R.sup.3 is --OR.sup.11
and R.sup.11 is the residue of a naturally occurring amino acid; Y
is O; R.sup.10 together with R.sup.9 form a double bond; R.sup.8 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0209] In another embodiment of Formula IV, R.sup.3 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; Y is O; R.sup.10
together with R.sup.9 form a double bond; R.sup.8 is absent; and
R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0210] In one embodiment of Formula IV, the dotted line between C4
and C5 is a single bond and the dotted line between C5 and C6 is a
single bond.
[0211] In another embodiment of Formula IV, the dotted line between
C4 and C5 is a single bond and the dotted line between C5 and C6 is
a double bond.
[0212] In another embodiment of Formula IV, the dotted line between
C4 and C5 is a double bond and the dotted line between C5 and C6 is
a single bond.
[0213] In still another embodiment of Formula IV, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0214] In one embodiment of Formula IV, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0215] In one embodiment of Formula IV, R.sup.3 comprises a residue
of a naturally occurring .alpha.-amino acid in the L-configuration.
In another embodiment, R.sup.3 comprises a residue of L-valine. In
another embodiment, R.sup.3 comprises an amino acid residue with
the D-configuration. In another embodiment, R.sup.3 comprises a
non-natural amino acid. In other embodiments, R.sup.3 comprises the
residue of a .beta. .gamma. or .delta. amino acid.
[0216] In one preferred embodiment of Formula IV, R.sup.3
represents an ester of an amino acid. In another embodiment,
R.sup.3 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid. In certain embodiments of Formula IV, a residue of
an amino acid is connected to the steroid ring system at the
carboxyl group of the amino acid. In other embodiments, a residue
of an amino acid is connected to the steroid at the amino acid side
chain. For example, amino acids that contain side chains with
functional groups that are capable of forming a bond with a hydroxy
or a ketone group may be boded to the steroid ring by such a group.
In other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid, arginine and the like.
[0217] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0218] In one embodiment, a compound of Formula IV with the
stereochemical configuration of Formula IVa is provided:
##STR00013##
[0219] wherein the definition of variables Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.5, R.sup.6, R.sup.8, R.sup.9 and R.sup.10 are as
defined in Formula IV above.
[0220] In another embodiment, a compound of Formula IV with the
stereochemical configuration of Formula IVb is provided:
##STR00014##
[0221] wherein the definition of variables Y, R.sup.1, R.sup.2,
R.sup.3, R.sup.5, R.sup.6, R.sup.8, R.sup.9 and R.sup.10 are as
defined in Formula IV above.
[0222] In another embodiment, a progesterone or steroid analogue of
Formula V is provided:
##STR00015##
[0223] or a pharmaceutically acceptable salt, ester or prodrug
thereof, wherein X is O, N or S;
[0224] R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0225] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid a carbohydrate; --OR.sup.11; --NR.sup.11R.sup.12,
monophosphate, diphosphate, triphosphate; --C(O)R', where R' is
alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue of
an amino acid; R.sup.3 together with X and R.sup.4 forms an
optionally substituted 5-6 membered heterocyclic or heteroaryl ring
containing 1-4 nitrogen, oxygen or sulfur atoms, or R.sup.3 is
absent;
[0226] R.sup.4 is hydrogen or alkyl; R.sup.4 together with R.sup.3
and X forms an optionally substituted 5-6 membered heterocyclic or
heteroaryl ring containing 1-4 nitrogen, oxygen or sulfur atoms, or
R.sup.4 together with R.sup.7 form a double bond;
[0227] R.sup.8 is --OR.sup.11, --NR.sup.11R.sup.12, monophosphate,
diphosphate, triphosphate, a carbohydrate, --C(O)R', where R' is
alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue of
an amino acid;
[0228] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, a
carbohydrate, monophosphate, diphosphate, triphosphate; --C(O)R',
where R' is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2, or the
residue of an amino acid; or an optionally substituted ester;
[0229] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond; and with the
proviso that XR.sup.3R.sup.7 does not represent an ester of
aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid; and with the proviso
that R.sup.8 does not represent aspartic acid, glutamic acid, gamma
amino butyric acid or .alpha.-2-(hydroxyethylamino)-propionic
acid.
[0230] In one embodiment of Formula V, R.sup.3 is hydrogen,
optionally substituted acyl, a residue of an amino acid a
carbohydrate; --OR.sup.11; --NR.sup.11R.sup.12, or R.sup.3 is
absent; R.sup.8 is --OR.sup.11, --NR.sup.11R.sup.12 or a
carbohydrate; R.sup.11 is the residue of an amino acid, a
carbohydrate or an optionally substituted ester.
[0231] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0232] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0233] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0234] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0235] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0236] In still another embodiment, R.sup.4 is hydrogen or
methyl.
[0237] In some embodiments, X is O. In other embodiments, X is N.
In certain embodiments in which X is O, R.sup.3 and R.sup.4 come
together to form a double bond. In certain embodiments, one of
R.sup.3 and R.sup.8 is a residue of an amino acid. In particular
embodiments, the amino acid is a naturally occurring amino acid. In
certain embodiments, R.sup.3 is a residue of an amino acid. In
certain other embodiments, R.sup.8 is a residue of an amino acid.
In yet further embodiments, both R.sup.3 and R.sup.8 are residues
of an amino acid.
[0238] In other embodiments, R.sup.3 is a carbohydrate. In another
embodiment, R.sup.8 is a carbohydrate.
[0239] In one embodiment of Formula V, X is O; R.sup.3 is the
residue of an amino acid or a carbohydrate; and R.sup.7 is
absent.
[0240] In another embodiment of Formula V, X is N; R.sup.4 together
with R.sup.7 form a double bond; R.sup.3 is OR.sup.11; and R.sup.11
is the residue of an amino acid or a carbohydrate.
[0241] In another embodiment of Formula V, X is N; R.sup.4 together
with R.sup.7 form a double bond; R.sup.3 is --NR.sup.11R.sup.12;
R.sup.11 is the residue of an amino acid or a carbohydrate; and
R.sup.12 is hydrogen.
[0242] In another embodiment of Formula V, R.sup.8 is --OR.sup.11;
R.sup.11 is the residue of a naturally occurring amino acid or a
carbohydrate; X is O; R.sup.4 together with R.sup.7 form a double
bond; and R.sup.3 is absent.
[0243] In another embodiment of Formula V, R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; X is O; R.sup.4
together with R.sup.7 form a double bond; and R.sup.3 is
absent.
[0244] In another embodiment of Formula V, R.sup.8 is --OR.sup.11;
R.sup.11 is a carbohydrate; X is O; R.sup.4 together with R.sup.7
form a double bond; and R.sup.3 is absent.
[0245] In another embodiment of Formula V, R.sup.8 is --OR.sup.11;
R.sup.11 is the residue of a naturally occurring amino acid or a
carbohydrate; X is O; R.sup.3 and R.sup.4 are hydrogen; and R.sup.7
is absent.
[0246] In another embodiment of Formula V, R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid or a carbohydrate; R.sup.12 is hydrogen; X is
O; R.sup.3 and R.sup.4 are hydrogen; and R.sup.7 is absent.
[0247] In another embodiment of Formula V, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula V, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula V, R.sup.1 is alkyl; and
R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of Formula
V, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are hydrogen. In still
another embodiment, of Formula V, R.sup.5 is alkyl; and R.sup.1 and
R.sup.2 are hydrogen.
[0248] In another embodiment of Formula V, R.sup.8 is --OR.sup.11;
R.sup.11 is the residue of a naturally occurring amino acid; X is
O; R.sup.4 together with R.sup.7 form a double bond; R.sup.3 is
absent; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0249] In another embodiment of Formula V, R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; X is O; R.sup.4
together with R.sup.7 form a double bond; R.sup.3 is absent; and
R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0250] In another embodiment of Formula V, R.sup.8 is --OR.sup.11;
R.sup.11 is the residue of a naturally occurring amino acid; X is
O; R.sup.3 and R.sup.4 are hydrogen; R.sup.7 is absent; and
R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0251] In another embodiment of Formula V, R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid; R.sup.12 is hydrogen; X is O; R.sup.3 and
R.sup.4 are hydrogen; R.sup.7 is absent; and R.sup.1, R.sup.2,
R.sup.5 and R.sup.6 are hydrogen; and R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are hydrogen.
[0252] In one embodiment of Formula V, the dotted line between C4
and C5 is a single bond and the dotted line between C5 and C6 is a
single bond.
[0253] In another embodiment of Formula V, the dotted line between
C4 and C5 is a single bond and the dotted line between C5 and C6 is
a double bond.
[0254] In another embodiment of Formula V, the dotted line between
C4 and C5 is a double bond and the dotted line between C5 and C6 is
a single bond.
[0255] In still another embodiment of Formula V, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0256] In another embodiment of Formula V, --XR.sup.3R.sup.7 is in
the alpha configuration. In still another embodiment,
--XR.sup.3R.sup.7 is in the beta configuration.
[0257] In one embodiment of Formula V, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, the hydrogen
at the C5 bridgehead carbon is in the alpha configuration. In
another embodiment, the dotted lines between C4 and C5 and between
C5 and C6 represent a single bond, the hydrogen at the C5
bridgehead carbon is in the beta configuration.
[0258] In one embodiment of Formula V, R.sup.8 comprises a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.8 comprises a residue of L-valine. In
another embodiment, R.sup.8 comprises an amino acid residue with
the D-configuration. In another embodiment, R.sup.8 comprises a
non-natural amino acid. In other embodiments, R.sup.8 comprises the
residue of a .beta. .gamma. or .delta. amino acid.
[0259] In one preferred embodiment of Formula V, R.sup.3 represents
an ester of an amino acid. In another embodiment, R.sup.3
represents an ester of an amino acid residue where the ester bond
is formed with a carboxylate group on the side chain of the amino
acid. In certain embodiments of Formula V, a residue of an amino
acid is connected to the steroid ring system at the carboxyl group
of the amino acid. In other embodiments, a residue of an amino acid
is connected to the steroid at the amino acid side chain. For
example, amino acids that contain side chains with functional
groups that are capable of forming a bond with a hydroxy or a
ketone group may be boded to the steroid ring by such a group. In
other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid, arginine and the like.
[0260] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0261] In one embodiment, a compound of Formula V with the
stereochemical configuration of Formula Va is provided:
##STR00016##
[0262] wherein the definition of variables X, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, and R.sup.8 are as
defined in Formula V above.
[0263] In another embodiment, a compound of Formula V with the
stereochemical configuration of Formula Vb is provided:
##STR00017##
[0264] wherein the definition of variables X, R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, and R.sup.8 are as
defined in Formula V above.
[0265] In another embodiment of the invention, a progesterone
analogue of Formula VI is provided:
##STR00018## [0266] or a pharmaceutically acceptable salt, ester or
prodrug thereof, wherein R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen, hydroxyl cycloalkyl,
cycloalkenyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl,
heterocyclic, heteroaryl, amino, thiol, alkoxy, sulfide, nitro,
cyano, azide, sulfonyl, acyl, carboxyl, an ester, an amide,
carbamate, carbonate, an amino acid residue or a carbohydrate;
[0267] R.sup.3 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate; --C(O)R', where R' is alkyl, aryl, aralkyl, OR.sup.1,
NR.sup.1R.sup.2 or the residue of an amino acid; or R.sup.3
together with the oxygen and R.sup.4 forms an optionally
substituted 5-6 membered heterocyclic or heteroaryl ring containing
1-4 nitrogen, oxygen or sulfur atoms;
[0268] R.sup.4 is hydrogen, alkyl or R.sup.4 together with R.sup.3
and the oxygen forms an optionally substituted 5-6 membered
heterocyclic or heteroaryl ring containing 1-4 nitrogen, oxygen or
sulfur atoms; and the dotted lines indicate the presence of either
a single bond or a double bond, wherein the valences of a single
bond are completed by hydrogens, provided that if the dotted line
between C4 and C5 or between C5 and C6 represents a double bond
then the other dotted line between C4 and C5 or between C5 and C6
represents a single bond; and with the proviso that neither R.sup.3
does not represent an ester of aspartic acid, glutamic acid, gamma
amino butyric acid or .alpha.-2-(hydroxyethylamino)-propionic
acid.
[0269] In one embodiment of Formula VI, R.sup.3 is hydrogen,
optionally substituted acyl, a residue of an amino acid or a
carbohydrate.
[0270] In another embodiment of Formula VI, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula VI, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula VI, R.sup.1 is alkyl; and
R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of Formula
VI, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are hydrogen. In
still another embodiment, of Formula VI, R.sup.5 is alkyl; and
R.sup.1 and R.sup.2 are hydrogen.
[0271] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0272] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0273] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0274] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0275] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0276] In still another embodiment, R.sup.4 is hydrogen or
methyl.
[0277] In another embodiment of Formula VI, R.sup.3 is the residue
of a naturally occurring amino acid; and R.sup.1, R.sup.2, R.sup.4,
R.sup.5 and R.sup.6 are hydrogen.
[0278] In another embodiment of Formula VI, R.sup.3 is the residue
of a naturally occurring amino acid or a carbohydrate; R.sup.4 is
alkyl; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0279] In one embodiment of Formula VI, R.sup.1 is alkyl; and
R.sup.2, R.sup.4 and R.sup.5 are hydrogen.
[0280] In another embodiment of Formula VI, R.sup.1 and R.sup.4 are
alkyl; and R.sup.2 and R.sup.5 are hydrogen.
[0281] In another embodiment of Formula VI, R.sup.3 is a
carbohydrate; and R.sup.1, R.sup.2, R.sup.4, R.sup.5 and R.sup.6
are hydrogen.
[0282] In another embodiment of Formula VI, R.sup.3 is --C(O)R',
where R' is alkyl, aryl, aralkyl, or the residue of an amino acid;
and R.sup.1, R.sup.2, R.sup.4, R.sup.5 and R.sup.6 are
hydrogen.
[0283] In one embodiment of Formula VI, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0284] In another embodiment of Formula VI, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0285] In another embodiment of Formula VI, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0286] In still another embodiment of Formula VI, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0287] In another embodiment of Formula VI, OR.sup.3 is in the
alpha configuration. In still another embodiment, OR.sup.3 is in
the beta configuration.
[0288] In one embodiment of Formula VI, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration.
[0289] In one embodiment of Formula VI, R.sup.3 represents a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.3 is a residue of L-valine. In another
embodiment, R.sup.3 represents an amino acid residue with the
D-configuration. In another embodiment, R.sup.3 represents a
non-natural amino acid. In other embodiments, R.sup.3 represents
the residue of a .beta. .gamma. or .delta. amino acid.
[0290] In one preferred embodiment of Formula VI, R.sup.3
represents an ester of an amino acid. In another embodiment,
R.sup.3 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid. In certain embodiments of Formula VI, a residue of
an amino acid is connected to the steroid ring system at the
carboxyl group of the amino acid. In other embodiments, a residue
of an amino acid is connected to the steroid at the amino acid side
chain. For example, amino acids that contain side chains with
functional groups that are capable of forming a bond with a hydroxy
or a ketone group may be bonded to the steroid ring by such a
group. In other embodiments, the reactive groups on the amino acid
side chains may displace leaving groups formed on the steroid
moiety to form a covalent bond. Non-limiting examples of amino
acids with reactive groups in the side chain include lysine,
cysteine, serine, tyrosine, aspartic acid, arginine and the
like.
[0291] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0292] In one embodiment, a compound of Formula VI with the
stereochemical configuration of Formula VIa is provided:
##STR00019##
[0293] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are as defined in Formula VI
above.
[0294] In another embodiment, a compound of Formula VI with the
stereochemical configuration of Formula VIb is provided:
##STR00020##
[0295] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are as defined in Formula VI
above.
[0296] In still another embodiment, a progesterone analogue of
Formula VII is provided:
##STR00021##
[0297] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0298] wherein, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen, hydroxyl cycloalkyl,
cycloalkenyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl,
heterocyclic, heteroaryl, amino, thiol, alkoxy, sulfide, nitro,
cyano, azide, sulfonyl, acyl, carboxyl, an ester, an amide,
carbamate, carbonate, an amino acid residue or a carbohydrate;
[0299] R.sup.3 is --OR.sup.11, --NR.sup.11R.sup.12, monophosphate,
diphosphate, triphosphate, a carbohydrate, or --C(O)R', where R' is
alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue of
an amino acid;
[0300] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, a
carbohydrate, monophosphate, diphosphate, triphosphate; --C(O)R',
where R' is OR.sup.1, NR.sup.1R.sup.2, alkyl, aryl, aralkyl, or the
residue of an amino acid; or an optionally substituted ester;
[0301] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond.
[0302] In one embodiment of Formula VII, R.sup.3 is --OR.sup.11,
--NR.sup.11R.sup.12, or a carbohydrate; and R.sup.11 is the residue
of an amino acid, a carbohydrate, or an optionally substituted
ester.
[0303] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0304] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0305] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0306] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0307] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0308] In one embodiment of Formula VII, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula VII, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula VII, R.sup.1 is alkyl;
and R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of
Formula VII, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are
hydrogen. In still another embodiment, of Formula VII, R.sup.5 is
alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0309] In another embodiment of Formula VII, R.sup.3 is --OR.sup.11
and R.sup.11 is the residue of a naturally occurring amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0310] In another embodiment of Formula VII, R.sup.3 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid or a carbohydrate; R.sup.12 is hydrogen; and
R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0311] In another embodiment of Formula VII, R.sup.3 OR.sup.11,
R.sup.11 is a carbohydrate; and R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are hydrogen.
[0312] In another embodiment of Formula VII, R.sup.3 is --C(O)R',
where R' is alkyl, aryl, aralkyl, or the residue of an amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0313] In one embodiment of Formula VII, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0314] In another embodiment of Formula VII, the dotted line
between C4 and C5 represents a single bond and the dotted line
between C5 and C6 represents a double bond.
[0315] In another embodiment of Formula VII, the dotted line
between C4 and C5 represents a double bond and the dotted line
between C5 and C6 represents a single bond.
[0316] In still another embodiment of Formula VII, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0317] In one embodiment of Formula VII, the dotted lines between
C4 and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0318] In one embodiment of Formula VII, R.sup.3 comprises a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.3 comprises a residue of L-valine. In
another embodiment, R.sup.3 comprises an amino acid residue with
the D-configuration. In another embodiment, R.sup.3 comprises a
non-natural amino acid. In other embodiments, R.sup.3 comprises the
residue of a .beta. .gamma. or .delta. amino acid.
[0319] In one preferred embodiment of Formula V, R.sup.3 represents
an ester of an amino acid. In another embodiment, R.sup.3
represents an ester of an amino acid residue where the ester bond
is formed with a carboxylate group on the side chain of the amino
acid. In certain embodiments of Formula V, a residue of an amino
acid is connected to the steroid ring system at the carboxyl group
of the amino acid. In other embodiments, a residue of an amino acid
is connected to the steroid at the amino acid side chain. For
example, amino acids that contain side chains with functional
groups that are capable of forming a bond with a hydroxy or a
ketone group may be boded to the steroid ring by such a group. In
other embodiments, the reactive groups on the amino acid side
chains may displace leaving groups formed on the steroid moiety to
form a covalent bond. Non-limiting examples of amino acids with
reactive groups in the side chain include lysine, cysteine, serine,
tyrosine, aspartic acid, arginine and the like.
[0320] The amino acid(s) in any of the embodiments of the invention
described herein may be naturally occurring or synthetic amino
acids and may be in the D or L stereoisomeric form or may exist as
a D, L mixture. For example the 20 naturally occurring
.alpha.-amino acids in the L-configuration are encompassed by the
invention as well as .alpha.-amino acids in the D-configuration.
Synthetic amino acids in either stereoisomeric form are also
encompassed.
[0321] In one embodiment, a compound of Formula VII with the
stereochemical configuration of Formula VIIa is provided:
##STR00022##
[0322] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.3, R.sup.5, R.sup.6, are as defined in Formula VII above.
[0323] In another embodiment, a compound of Formula VII with the
stereochemical configuration of Formula VIIb is provided:
##STR00023##
[0324] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5, and R.sup.6 are as defined in Formula
VII above.
[0325] In another embodiment of the invention, a progesterone
analogue of Formula VIII is provided:
##STR00024##
[0326] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0327] wherein R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen, hydroxyl cycloalkyl,
cycloalkenyl, alkenyl, alkynyl, aryl, alkylaryl, arylalkyl,
heterocyclic, heteroaryl, amino, thiol, alkoxy, sulfide, nitro,
cyano, azide, sulfonyl, acyl, carboxyl, an ester, an amide,
carbamate, carbonate, an amino acid residue or a carbohydrate;
[0328] R.sup.8 is hydrogen, optionally substituted acyl, a residue
of an amino acid, a carbohydrate, monophosphate, diphosphate,
triphosphate; --C(O)R', where R' is alkyl, aryl, aralkyl, OR.sup.1,
NR.sup.1R.sup.2 or the residue of an amino acid; or, R.sup.8
together with R.sup.9 and the oxygen forms an optionally
substituted 5-6 membered heterocyclic or heteroaryl ring containing
1-4 nitrogen, oxygen or sulfur atoms;
[0329] R.sup.9 is hydrogen, alkyl, or R.sup.9 together with the
oxygen and R.sup.8 forms an optionally substituted 5-6 membered
heterocyclic or heteroaryl ring containing 1-4 nitrogen, oxygen or
sulfur atoms; and the dotted lines indicate the presence of either
a single bond or a double bond, wherein the valences of a single
bond are completed by hydrogens, provided that if the dotted line
between C4 and C5 or between C5 and C6 represents a double bond
then the other dotted line between C4 and C5 or between C5 and C6
represents a single bond; and with the proviso that R.sup.8 does
not represent an ester of aspartic acid, glutamic acid, gamma amino
butyric acid or .alpha.-2-(hydroxyethylamino)-propionic acid; and
with the proviso that when Y is N, R.sup.8 does not represent
aspartic acid, glutamic acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0330] In one embodiment of Formula VIII, R.sup.8 is hydrogen,
optionally substituted acyl, a residue of an amino acid or a
carbohydrate.
[0331] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0332] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0333] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0334] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0335] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0336] In still another embodiment, R.sup.9 is hydrogen or
methyl.
[0337] In another embodiment of Formula VIII, R.sup.6 is alkyl or
fluoro. In yet another embodiment of Formula VIII, R.sup.1, R.sup.2
and R.sup.5 are independently hydrogen or alkyl. In another
embodiment, R.sup.1 and R.sup.2 are hydroxyl. In still another
embodiment, R.sup.1 and R.sup.2 are independently hydroxyl or
halogen. In another embodiment of Formula VIII, R.sup.1 is alkyl;
and R.sup.2 and R.sup.5 are hydrogen. In another embodiment, of
Formula VIII, R.sup.2 is alkyl; and R.sup.1 and R.sup.5 are
hydrogen. In still another embodiment, of Formula VIII, R.sup.5 is
alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0338] In another embodiment of Formula VIII, R.sup.8 is the
residue of a naturally occurring amino acid; and R.sup.1, R.sup.2,
R.sup.5, R.sup.6 and R.sup.9 are hydrogen.
[0339] In another embodiment of Formula VIII, R.sup.8 is the
residue of a naturally occurring amino acid or a carbohydrate;
R.sup.9 is alkyl; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
hydrogen.
[0340] In one embodiment of Formula VIII, R.sup.1 is alkyl; and
R.sup.2, R.sup.5 and R.sup.9 are hydrogen.
[0341] In another embodiment of Formula VIII, R.sup.1 and R.sup.9
are alkyl; and R.sup.2 and R.sup.5 are hydrogen.
[0342] In another embodiment of Formula VIII, R.sup.8 is a
carbohydrate; and R.sup.1, R.sup.2, R.sup.5, R.sup.6 and R.sup.9
are hydrogen.
[0343] In another embodiment of Formula VIII, R.sup.8 is --C(O)R',
where R' is alkyl, aryl, aralkyl, or the residue of an amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0344] In one embodiment of Formula VIII, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a single bond.
[0345] In another embodiment of Formula VIII, the dotted line
between C4 and C5 represents a single bond and the dotted line
between C5 and C6 represents a double bond.
[0346] In another embodiment of Formula VIII, the dotted line
between C4 and C5 represents a double bond and the dotted line
between C5 and C6 represents a single bond.
[0347] In still another embodiment of Formula VIII, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0348] In one embodiment of Formula VIII, the dotted lines between
C4 and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0349] In one embodiment of Formula VIII, R.sup.8 is the residue of
a naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.8 is a residue of L-valine. In another
embodiment, R.sup.8 is an amino acid residue with the
D-configuration. In another embodiment, R.sup.8 represents a
residue of a non-natural amino acid. In other embodiments, R.sup.8
represents the residue of a .beta. .gamma. or .delta. amino
acid.
[0350] In one preferred embodiment of Formula VIII, R.sup.8
represents an ester of an amino acid. In another embodiment,
R.sup.8 represents an ester of an amino acid residue where the
ester bond is formed with a carboxylate group on the side chain of
the amino acid.
[0351] In one embodiment, a compound of Formula VIII with the
stereochemical configuration of Formula VIIIa is provided:
##STR00025##
[0352] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, R.sup.6, R.sup.8, and R.sup.9 are as defined in Formula
VIII above.
[0353] In another embodiment, a compound of Formula VIII with the
stereochemical configuration of Formula VIIIb is provided:
##STR00026##
[0354] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, R.sup.6, R.sup.8 and R.sup.9 are as defined in Formula
VIII above.
[0355] In still another embodiment of the invention, a steroid
analogue of Formula IX is provided:
##STR00027##
[0356] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate;
[0357] R.sup.8 is --OR.sup.11, --NR.sup.11R.sup.12, --C(O)R', where
R' is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the
residue of an amino acid; or a carbohydrate;
[0358] R.sup.11 is hydrogen, optionally substituted alkyl,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, a
carbohydrate or an optionally substituted ester; --C(O)R', where R'
is alkyl, aryl, aralkyl, OR.sup.1, NR.sup.1R.sup.2 or the residue
of an amino acid;
[0359] R.sup.12 is hydrogen or alkyl; and the dotted lines indicate
the presence of either a single bond or a double bond, wherein the
valences of a single bond are completed by hydrogens, provided that
if the dotted line between C4 and C5 or between C5 and C6
represents a double bond then the other dotted line between C4 and
C5 or between C5 and C6 represents a single bond; and with the
proviso that R.sup.8 does not represent aspartic acid, glutamic
acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0360] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0361] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0362] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0363] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0364] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0365] In one embodiment of Formula IX, R.sup.6 is alkyl or fluoro.
In yet another embodiment of Formula IX, R.sup.1, R.sup.2 and
R.sup.5 are independently hydrogen or alkyl. In another embodiment,
R.sup.1 and R.sup.2 are hydroxyl. In still another embodiment,
R.sup.1 and R.sup.2 are independently hydroxyl or halogen. In
another embodiment of Formula IX, R.sup.1 is alkyl; and R.sup.2 and
R.sup.5 are hydrogen. In another embodiment, of Formula IX, R.sup.2
is alkyl; and R.sup.1 and R.sup.5 are hydrogen. In still another
embodiment, of Formula IX, R.sup.5 is alkyl; and R.sup.1 and
R.sup.2 are hydrogen.
[0366] In another embodiment of Formula IX, R.sup.3 is --OR.sup.11
and R.sup.11 is the residue of a naturally occurring amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0367] In another embodiment of Formula IX, R.sup.8 is
--NR.sup.11R.sup.12; R.sup.11 is the residue of a naturally
occurring amino acid or a carbohydrate; R.sup.12 is hydrogen; and
R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0368] In another embodiment of Formula IX, R.sup.8 is a
carbohydrate; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
hydrogen.
[0369] In another embodiment of Formula IX, R.sup.8 is --C(O)R',
where R' is alkyl, aryl, aralkyl, or the residue of an amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are hydrogen.
[0370] In one embodiment of Formula IX, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0371] In another embodiment of Formula IX, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0372] In another embodiment of Formula IX, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0373] In still another embodiment of Formula IX, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0374] In one embodiment of Formula IX, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, and the hydrogen at the
C5 bridgehead carbon is in the beta configuration
[0375] In one embodiment of Formula IX, R.sup.8 comprises a
naturally occurring .alpha.-amino acid in the L-configuration. In
another embodiment, R.sup.8 comprises a residue of L-valine. In
another embodiment, R.sup.8 comprises an amino acid residue with
the D-configuration. In another embodiment, R.sup.8 comprises a
non-natural amino acid. In other embodiments, R.sup.8 comprises the
residue of a .beta. .gamma. or .delta. amino acid.
[0376] In one embodiment, a compound of Formula IX with the
stereochemical configuration of Formula IXa is provided:
##STR00028##
[0377] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, R.sup.6, and R.sup.8 are as defined in Formula IX
above.
[0378] In another embodiment, a compound of Formula IX with the
stereochemical configuration of Formula IXb is provided:
##STR00029##
[0379] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, R.sup.6, and R.sup.8 are as defined in Formula IX
above.
[0380] In still another embodiment of the invention, a steroid
analogue of Formula X is provided:
##STR00030##
[0381] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
wherein R is the side chain of a naturally occurring amino acid;
and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate; and with the proviso that
R does not represent the side chain of aspartic acid, glutamic
acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0382] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0383] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0384] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0385] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0386] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0387] In one embodiment of Formula X, R.sup.6 is alkyl or fluoro.
In yet another embodiment of Formula X, R.sup.1, R.sup.2 and
R.sup.5 are independently hydrogen or alkyl. In another embodiment,
R.sup.1 and R.sup.2 are hydroxyl. In still another embodiment,
R.sup.1 and R.sup.2 are independently hydroxyl or halogen. In
another embodiment of Formula X, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are hydrogen. In one embodiment of Formula X, R.sup.1 is
alkyl; and R.sup.2, and R.sup.5 are hydrogen. In another
embodiment, of Formula X, R.sup.2 is alkyl; and R.sup.1 and R.sup.5
are hydrogen. In still another embodiment, of Formula X, R.sup.5 is
alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0388] In one embodiment of Formula X, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0389] In another embodiment of Formula X, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0390] In another embodiment of Formula X, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0391] In still another embodiment of Formula X, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0392] In one embodiment of Formula X, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, and the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, the hydrogen at the C5
bridgehead carbon is in the beta configuration
[0393] In one embodiment of Formula X, R comprises the side chain
of a naturally occurring .alpha.-amino acid in the L-configuration.
In another embodiment, R comprises a residue of L-alanine,
L-leucine, L-isoleucine, L-proline, L-proline, L-phenylalanine,
L-tryptophan, L-lysine, or L-valine. In another embodiment, R
comprises an amino acid residue with the D-configuration.
[0394] In one embodiment, a compound of Formula X with the
stereochemical configuration of Formula Xa is provided:
##STR00031##
[0395] wherein the definition of variables R, R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula X above.
[0396] In another embodiment, a compound of Formula X with the
stereochemical configuration of Formula Xb is provided:
##STR00032##
[0397] wherein the definition of variables R, R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula X above.
[0398] In still another embodiment of the invention, a steroid
analogue of Formula XI is provided:
##STR00033##
[0399] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0400] where R is the side chain of a naturally occurring amino
acid; and R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are independently
hydrogen, alkyl, halogen, hydroxyl cycloalkyl, cycloalkenyl,
alkenyl, alkynyl, aryl, alkylaryl, arylalkyl, heterocyclic,
heteroaryl, amino, thiol, alkoxy, sulfide, nitro, cyano, azide,
sulfonyl, acyl, carboxyl, an ester, an amide, carbamate, carbonate,
an amino acid residue or a carbohydrate; and with the proviso that
R does not represent the side chain of aspartic acid, glutamic
acid, gamma amino butyric acid or
.alpha.-2-(hydroxyethylamino)-propionic acid.
[0401] In one embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6 are
independently hydrogen, alkyl, halogen or hydroxyl.
[0402] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently methyl, ethyl or propyl.
[0403] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently thiomethyl, hydroxymethyl or cyano.
[0404] In another embodiment, R.sup.1, R.sup.2, R.sup.5 and R.sup.6
are independently vinyl or ethynyl.
[0405] In still another embodiment, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are independently fluoro, bromo, chloro or iodo.
[0406] In one embodiment of Formula X, R.sup.6 is alkyl or fluoro.
In yet another embodiment of Formula X, R.sup.1, R.sup.2 and
R.sup.5 are independently hydrogen or alkyl. In another embodiment,
R.sup.1 and R.sup.2 are hydroxyl. In still another embodiment,
R.sup.1 and R.sup.2 are independently hydroxyl or halogen. In
another embodiment of Formula X, R.sup.1, R.sup.2, R.sup.5 and
R.sup.6 are hydrogen. In one embodiment of Formula X, R.sup.1 is
alkyl; and R.sup.2, and R.sup.5 are hydrogen. In another
embodiment, of Formula X, R.sup.2 is alkyl; and R.sup.1 and R.sup.5
are hydrogen. In still another embodiment, of Formula X, R.sup.5 is
alkyl; and R.sup.1 and R.sup.2 are hydrogen.
[0407] In one embodiment of Formula X, the dotted line between C4
and C5 represents a single bond and the dotted line between C5 and
C6 represents a single bond.
[0408] In another embodiment of Formula X, the dotted line between
C4 and C5 represents a single bond and the dotted line between C5
and C6 represents a double bond.
[0409] In another embodiment of Formula X, the dotted line between
C4 and C5 represents a double bond and the dotted line between C5
and C6 represents a single bond.
[0410] In still another embodiment of Formula X, the dotted line
between C1 and C2 represents a single bond. In still another
embodiment, the dotted line between C1 and C2 represents a double
bond.
[0411] In one embodiment of Formula X, the dotted lines between C4
and C5 and between C5 and C6 represent a single bond, and the
hydrogen at the C5 bridgehead carbon is in the alpha configuration.
In another embodiment, and the dotted lines between C4 and C5 and
between C5 and C6 represent a single bond, the hydrogen at the C5
bridgehead carbon is in the beta configuration
[0412] In one embodiment of Formula X, R comprises the side chain
of a naturally occurring .alpha.-amino acid in the L-configuration.
In another embodiment, R comprises a residue of L-alanine,
L-leucine, L-isoleucine, L-proline, L-proline, L-phenylalanine,
L-tryptophan, L-lysine, or L-valine. In another embodiment, R
comprises an amino acid residue with the D-configuration.
[0413] In one embodiment, a compound of Formula XI with the
stereochemical configuration of Formula XIa is provided:
##STR00034##
[0414] wherein the definition of variables R, R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula XI above.
[0415] In another embodiment, a compound of Formula XI with the
stereochemical configuration of Formula XIb is provided:
##STR00035##
[0416] wherein the definition of variables R, R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula XI above.
[0417] In another embodiment, a steroid analogue of Formula XII is
provided:
##STR00036##
[0418] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0419] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula I; and R.sup.13 and
R.sup.14 are hydrogen, optionally substituted alkyl, carboxy,
sulfonyl, sulfonic acid, sulfonamide, an ester, and amide,
hydroxyalkyl, aminoalkyl, the residue of an amino acid, or a
carbohydrate.
[0420] In another embodiment, a compound of Formula XII with the
stereochemical configuration of Formula XIIa is provided:
##STR00037##
[0421] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.13 and R.sup.14 are as defined for Formula XII.
[0422] In another embodiment, a compound of Formula XII with the
stereochemical configuration of Formula XIIb is provided:
##STR00038##
[0423] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.13 and R.sup.14 are as defined for Formula XII.
[0424] In still another embodiment of the invention, a steroid
analogue of Formula XIII is provided:
##STR00039##
[0425] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0426] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula I; and R.sup.13 and
R.sup.14 are independently selected from hydrogen, optionally
substituted alkyl, hydroxy, alkoxy, amino, alkylamino,
dialkylamino, carboxy, sulfonyl, sulfonic acid, sulfonamide, an
ester, and amide, hydroxyalkyl, aminoalkyl, alkylaminoalkyl,
dialkylaminoalkyl, the residue of an amino acid, or a
carbohydrate.
[0427] In some embodiments of Formula XIII, variables R.sup.1,
R.sup.2, R.sup.5, and R.sup.6, are as defined in Formula I; and
R.sup.13 and R.sup.14 are independently selected from hydrogen,
optionally substituted alkyl, carboxy, sulfonyl, sulfonic acid,
sulfonamide, an ester, and amide, hydroxyalkyl, aminoalkyl,
alkylaminoalkyl, dialkylaminoalkyl, the residue of an amino acid,
or a carbohydrate.
[0428] In another embodiment, a compound of Formula XIII with the
stereochemical configuration of Formula XIIIa is provided:
##STR00040##
[0429] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.13 and R.sup.14 are as defined for Formula XIII.
[0430] In another embodiment, a compound of Formula XIII with the
stereochemical configuration of Formula XIIIb is provided:
##STR00041##
[0431] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.3 and R.sup.14 are as defined for Formula XIII.
[0432] In still another embodiment, a compound of Formula XIV is
provided:
##STR00042##
[0433] or a pharmaceutically acceptable salt, ester or prodrug
thereof,
[0434] wherein the definition of variables R.sup.1, R.sup.2,
R.sup.5, and R.sup.6, are as defined in Formula I; and R.sup.13 and
R.sup.14 are hydrogen, optionally substituted alkyl, carboxy,
sulfonyl, sulfonic acid, sulfonamide, an ester, and amide,
hydroxyalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, the
residue of an amino acid, or a carbohydrate.
[0435] In another embodiment, a compound of Formula XIV with the
stereochemical configuration of Formula XIVa is provided:
##STR00043##
[0436] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.13 and R.sup.14 are as defined for Formula XIV.
[0437] In another embodiment, a compound of Formula XIV with the
stereochemical configuration of Formula XIVb is provided:
##STR00044##
[0438] wherein variables R.sup.1, R.sup.2, R.sup.5, R.sup.6,
R.sup.13 and R.sup.14 are as defined for Formula XIV.
[0439] In particular embodiments of the invention, the invention
provides the steroid analogues having the Formulas presented in
Table 1 below, or a pharmaceutically acceptable salts, esters or
prodrugs thereof.
TABLE-US-00001 TABLE 1 Compound # Structure P1-31 ##STR00045##
P1-57 ##STR00046## P1-79 ##STR00047## P1-113 ##STR00048## P1-123
##STR00049## P1-131 ##STR00050## P1-133 ##STR00051## P1-135
##STR00052## P1-29 ##STR00053## P1-32 ##STR00054## P1-33
##STR00055## P1-34 ##STR00056## P1-163 ##STR00057## P1-185
##STR00058## P1-186 ##STR00059## P2-29-E ##STR00060## P2-29-Z
##STR00061## P2-13 ##STR00062## ##STR00063## ##STR00064##
##STR00065## ##STR00066## ##STR00067## ##STR00068## ##STR00069##
##STR00070## ##STR00071## ##STR00072## ##STR00073## ##STR00074##
##STR00075## ##STR00076## ##STR00077## ##STR00078## ##STR00079##
##STR00080## ##STR00081## ##STR00082## ##STR00083##
##STR00084##
Stereochemistry
[0440] It is understood that based on the number of asymmetric
centers, a total number of 2.sup.n possible stereochemical isomers
is possible. The present invention includes all possible
stereochemical configurations of the compounds.
[0441] In some embodiments the stereochemistry of the compounds of
the invention will retain the natural stereochemistry of the
natural steroid. For example, the stereochemistry at C8, C9, C10,
C13, C14 and C17 will retain the stereochemistry of the natural
steroid compounds. In contrast, the compounds of the invention
include compounds with variable configurations at C-3 and C-5 of
the steroid ring system. In some embodiments, the configuration of
C-3 is alpha. In other embodiments, the configuration of C-3 is
beta. Similarly, in some embodiments, the configuration of C-5 is
alpha, and in other embodiments the configuration at C-5 is beta.
All possible combinations of stereochemical configurations at C-3
and C-5 are embraced by the invention.
[0442] Unless otherwise indicated, the stereochemistry of the
compounds of the invention will retain the natural stereochemistry
of progesterone at the bridgehead carbon atoms C-8, C-9,C-14 and
C-17. In addition, the stereochemistry of the quaternary carbons
C-10 and C-13 will also retain the stereochemistry of the
progesterone, unless indicated otherwise. In contrast, the
compounds of the invention include compounds with variable
configurations at C-3 and C-5 of the steroid ring system. In some
embodiments, the configuration of C-3 is alpha. In other
embodiments, the configuration of C-3 is beta. Similarly, in some
embodiments, the configuration of C-5 is alpha, and in other
embodiments the configuration at C-5 is beta. All possible
combinations of stereochemical configurations at C-3 and C-5 are
embraced by the invention.
[0443] In one embodiment of the invention, the pure E- or Z-isomers
of the carbonyl-derivatives of the steroid compounds, such as
oximes derivatives and the like, are provided. In another
embodiment, the invention provides mixtures of E- and Z-isomers of
the carbonyl derivatives of the neuroprotective compounds.
[0444] The present invention also encompasses all possible
stereochemical configurations of asymmetric substituents, such as
amino acids. As described above, the naturally occurring a-amino
acids in L, D, and D,L configurations are encompassed. Furthermore,
all possible stereochemical configurations of non-natural synthetic
amino acids are encompassed by the invention.
II. Definitions
[0445] It should be understood that the various possible
stereoisomers of the groups mentioned above and herein are within
the meaning of the individual terms and examples, unless otherwise
specified. As an illustrative example, "1-methyl-butyl" exists in
both the (R) and the (S) form, thus, both (R)-1-methyl-butyl and
(S)-1-methyl-butyl is covered by the term "1-methyl-butyl," unless
otherwise specified. Several biological compounds are designed by
the (D) and the (L) form, rather than the (R) and the (S) form,
respectively. As an another illustrative example, "glycine" exists
in both the (D) and the (L) form; therefore, both (D)-glycine and
(L)-glycine are covered by the term "glycine" unless otherwise
specified.
[0446] As used herein, the term "isolated enantiomer" refers to a
composition that includes at least approximately 95% to 100%, or
more preferably, over 97% of a single enantiomer of that
compound.
[0447] As used herein, the term "substantially free of" or
"substantially in the absence of" refers to a composition that
includes at least 85 or 90% by weight, preferably 95% to 98% by
weight, and even more preferably 99% to 100% by weight, of the
designated enantiomer of that compound.
[0448] The term "independently" is used herein to indicate that the
variable that is independently applied varies independently from
application to application. Thus, in a compound such as R''XYR'',
wherein R'' is "independently carbon or nitrogen," both R'' can be
carbon, both R'' can be nitrogen, or one R'' can be carbon and the
other R'' nitrogen.
[0449] Whenever a range is referred to herein, the range
independently includes every element thereof. For example, the term
"C.sub.1-5 alkyl" independently includes C.sub.1-alkyl,
C.sub.2-alkyl, C.sub.3-alkyl, C.sub.4-alkyl, C.sub.5-alkyl.
[0450] The term "alkyl," as used herein unless otherwise specified,
is intended to have its customary meaning in the art and includes
optionally substituted saturated straight, branched, or cyclic,
primary, secondary, or tertiary hydrocarbons. Alkyl, typically is
C.sub.1-16 alkyl but can also be lower alkyl, including C.sub.1-10
or C.sub.1-5 alkyl. In exemplary embodiments, alkyl includes
methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl,
cyclopentyl, isopentyl, neopentyl, hexylisohexyl, cyclohexyl,
cyclohexylmethyl, 3-methylpentyl, 2, 2-dimethylbutyl and
2,3-dimethylbutyl. When the alkyl group is a cyclic group, it is
typically between C.sub.3-12 or between C.sub.3-8 and can include
one or more cycles. The alkyl group can be optionally substituted
with one or more moieties. Examples of suitable substituents
include alkyl, halo, haloalkyl, hydroxyl, carboxyl, acyl, acyloxy,
amino, amido, carboxyl derivatives, alkylamino, dialkylamino,
arylamino, alkoxy, aryloxy, nitro, cyano, thiol, imine, sulfonic
acid, sulfate, sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester,
carboxylic acid, amide, phosphonyl, phosphinyl, phosphoryl,
phosphine, thioester, thioether, acid halide, anhydride, oxime,
hydrozine, carbamate, phosphonic acid, phosphate, phosphonate, or
any other viable functional group that does not inhibit the
pharmacological activity of this compound, either unprotected, or
protected as necessary, as known to those skilled in the art, for
example, as taught in Greene et al., "Protective Groups in Organic
Synthesis," John Wiley and Sons, 4.sup.th edition, 2007.
[0451] The term "protected" as used herein and unless otherwise
defined refers to a group that is added to an oxygen, nitrogen,
sulfur or phosphorus atom to prevent its further reaction or for
other purposes. A wide variety of oxygen and nitrogen protecting
groups are known to those skilled in the art or organic synthesis.
Suitable protecting groups are described, for example, in Greene,
et al., "Protective Groups in Organic Synthesis," John Wiley and
Sons, 4.sup.th edition, 2007.
[0452] The term "aryl," as used herein, is intended to have its
customary meaning in the art and includes, for example, phenyl,
biphenyl, and naphthyl and the like. The aryl group can be
optionally substituted. Non-limiting examples of substituents
include hydroxyl, amino, amido, alkylamino, dialkylamino,
haloalkyl, arylamino, alkoxy, aryloxy, halo, nitro, cyano, sulfonic
acid, thiol, imine, sulfonyl, sulfanyl, fulfinyl, fulfamonyl,
ester, sulfate, phosphonic acid, phosphate, phosphonyl, phosphinyl,
phosphoryl, phosphonate, phosphine, thioester, thioether, acid
halide, anhydride, oxime, hydrozine, carbamate or carboxyl, either
unprotected, or protected as necessary, as known to those skilled
in the art, for example, as taught in Greene, et al., "Protective
Groups in Organic Synthesis," John Wiley and Sons, 4.sup.th
edition, 2007.
[0453] The term "aralkyl," as used herein, and unless otherwise
specified, refers to an optionally substituted aryl group as
defined above linked to the molecule through an alkyl group as
defined above. The term alkaryl or alkylaryl as used herein, and
unless otherwise specified, refers to an alkyl group as defined
above linked to the molecule through an aryl group as defined
above. In each of these groups, the alkyl group can be optionally
substituted as describe above and the aryl group can be optionally
substituted as described above or with any other viable functional
group that does not inhibit the pharmacological activity of this
compound, either unprotected, or protected as necessary, as known
to those skilled in the art, for example, as taught in Greene, et
al., "Protective Groups in Organic Synthesis," John Wiley and Sons,
4.sup.th edition, 2007. Specifically included within the scope of
the term aryl are phenyl; naphthyl; phenylmethyl; phenylethyl;
3,4,5-trihydroxyphenyl; 3,4,5-trimethoxyphenyl;
3,4,5-triethoxyphenyl; 4-chlorophenyl; 4-methylphenyl;
3,5-di-tertiarybutyl-4-hydroxyphenyl; 4-fluorophenyl;
4-chloro-1-naphthyl; 2-methyl-1-naphthylmethyl; 2-naphthylmethyl;
4-chlorophenylmethyl; 4-tertiarybutylphenyl;
4-tertiarybutylphenylmethyl and the like.
[0454] The term "halo" or "halogen," as used herein includes
chloro, bromo, iodo and fluoro.
[0455] The term "heteroatom," as used herein, refers to oxygen,
sulfur, nitrogen or phosphorus.
[0456] The term "alkylamino" or "arylamino" refers to an amino
group that has one or two alkyl or aryl substituents,
respectively.
[0457] The term "alkoxy," as used herein, and unless otherwise
specified, refers to a moiety of the structure --O-alkyl, wherein
alkyl is as defined above.
[0458] The term "acyl" refers to moiety of the Formula --C(O)R',
wherein R' is alkyl, aryl, alkaryl, aralkyl, heteroaromatic,
heterocyclic, alkoxyalkyl including methoxymethyl, arylalkyl
including benzyl, aryloxyalkyl, such as phenoxymethyl, aryl
including optionally substituted phenyl.
[0459] As used herein, a "leaving group" means a functional group
that is cleaved from the molecule to which it is attached under
appropriate conditions.
[0460] The term "heteroaryl" or "heteroaromatic," as used herein
are intended to have their customary meaning in the art, and
include an aromatic group that includes at least one sulfur,
oxygen, nitrogen or phosphorus in the aromatic ring. The term
"heterocyclic" refers to a nonaromatic cyclic group wherein there
is at least one heteroatom, such as oxygen, sulfur, nitrogen or
phosphorus in the ring. Nonlimiting examples of heteroaryl and
heterocyclic groups include furyl, furanyl, pyridyl, pyrimidyl,
thienyl, isothiazolyl, imidazolyl, tetrazolyl, pyrazinyl,
benzofuranyl, benzothiophenyl, quinolyl, isoquinolyl, benzothienyl,
isobenzofuryl, pyrazolyl, indolyl, isoindolyl, benzimidazolyl,
purinyl, carbazolyl, oxazolyl, thiazolyl, isothiazolyl,
1,2,4-thiadiazolyl, isooxazolyl, pyrrolyl, quinazolinyl,
cinnolinyl, phthalazinyl, xanthinyl, hypoxanthinyl, thiophene,
furan, pyrrole, isopyrrole, pyrazole, or imidazole. The
heteroaromatic group can be optionally substituted as described
above for aryl. The heterocyclic group can be optionally
substituted with one or more moieties. Non-limiting examples of
suitable substituents include alkyl, halo, haloalkyl, hydroxyl,
carboxyl, acyl, acyloxy, amino, amido, carboxyl derivatives,
alkylamino, dialkylamino, arylamino, alkoxy, aryloxy, nitro, cyano,
sulfonic acid, thiol, imine, sulfonyl, sulfanyl, sulfinyl,
sulfamonyl, ester, carboxylic acid, amide, phosphonyl, phosphinyl,
phosphoryl, phosphine, thioester, thioether, acid halide,
anhydride, oxime, hydrozine, carbamate, phosphonic acid,
phosphonate, or any other viable functional group that does not
inhibit the pharmacological activity of this compound, either
unprotected, or protected as necessary, as known to those skilled
in the art, for example, as taught in Greene, et al., "Protective
Groups in Organic Synthesis," John Wiley and Sons, 4.sup.th
edition, 2007. The heteroaromatic can be partially or totally
hydrogenated as desired. As a non-limiting example, dihydropyridine
can be used in place of pyridine. Functional oxygen and nitrogen
groups on the heteroaryl group can be protected as necessary or
desired. Suitable protecting groups are well known to those skilled
in the art, and include, but are not limited to,
9-fluorenylmethoxycarbonyl (Fmoc), benzyl, trimethylsilyl,
dimethylhexylsilyl, t-butyldimethylsilyl, and t-butyldiphenylsilyl,
trityl or substituted trityl, alkyl groups, acyl groups such as
acetyl, benzoyl, and propionyl, methanesulfonyl, and
p-toluenesulfonyl.
[0461] Unless otherwise specified, the term "amino acid" includes
naturally occurring and synthetic .alpha., .beta. .gamma. or
.delta. amino acids. The naturally occurring amino acids are
glycine, alanine, valine, leucine, isoleucine, methionine,
phenylalanine, tryptophan, proline, serine, threonine, cysteine,
tyrosine, asparagine, glutamine, aspartate, glutamate, lysine,
arginine and histidine.
[0462] The term "residue of an amino acid" is intended to mean that
an amino acid is bonded to the compound at any substitutable
position, including for example, through an ester bond to the
carboxyl group of the amino acid. Other substitutable positions
include the amino group or the sidechain of the amino acid.
[0463] The term "side chain of a naturally occurring amino acid"
refers to the side chains of the 20 naturally occurring amino
acids, including hydrogen (i.e. the sidechain of glycine). The side
chains of the naturally occurring amino acids are shown below:
TABLE-US-00002 Alanine A, Ala Arginine R, Arg Asparagine N, Asn
Aspartic acid D, Asp Cysteine C, Cys Glutamine Q, Gln Glutamic acid
E, Glu Glycine G, Gly Histidine H, His Isoleucine I, Ile CH.sub.3--
Leucine HN.dbd.C(NH.sub.2)--NH--(CH.sub.2).sub.3-- Lysine
H.sub.2N--CO--CH.sub.2-- Methionine HOOC--CH.sub.2-- Phenylalanine
HS--CH.sub.2-- Proline H.sub.2N--CO--(CH.sub.2).sub.2-- Serine
HOOC--(CH.sub.2).sub.2-- Threonine H-- Tryptophan ##STR00085##
Tyrosine CH.sub.3--CH.sub.2--CH(CH.sub.3)-- Valine L, Leu
(CH.sub.3).sub.2--CH--CH.sub.2-- K, Lys
H.sub.2N--(CH.sub.2).sub.4-- M, Met CH.sub.3--S--(CH.sub.2).sub.2--
F, Phe Phenyl-CH.sub.2-- P, Pro ##STR00086## S, Ser HO--CH.sub.2--
T, Thr CH.sub.3--CH(OH)-- W, Trp ##STR00087## Y, Tyr
4-OH-Phenyl-CH.sub.2-- V, Val CH.sub.3--CH(CH.sub.2)--
[0464] In certain embodiments, the amino acid is in the
L-configuration. Alternatively, the amino acid can be a derivative
of alanyl, valinyl, leucinyl, isoleuccinyl, prolinyl,
phenylalaninyl, tryptophanyl, methioninyl, glycinyl, serinyl,
threoninyl, cysteinyl, tyrosinyl, asparaginyl, glutaminyl,
aspartoyl, glutaroyl, lysinyl, argininyl, histidinyl,
.beta.-alanyl, .beta.-valinyl, .beta.-leucinyl,
.beta.-isoleuccinyl, .beta.-prolinyl, .beta.-phenylalaninyl,
.beta.-tryptophanyl, .beta.-methioninyl, .beta.-glycinyl,
.beta.-serinyl, .beta.-threoninyl, .beta.-cysteinyl,
.beta.-tyrosinyl, .beta.-asparaginyl, .beta.-glutaminyl,
.beta.-aspartoyl, .beta.-glutaroyl, .beta.-lysinyl,
.beta.-argininyl or .beta.-histidinyl. When the term amino acid is
used, it is considered to be a specific and independent disclosure
of each of the esters of .alpha., .beta. .gamma. or .delta.
glycine, alanine, valine, leucine, isoleucine, methionine,
phenylalanine, tryptophan, proline, serine, threonine, cysteine,
tyrosine, asparagine, glutamine, aspartate, glutamate, lysine,
arginine and histidine in the D and L-configurations.
[0465] The term "thio" refers to a sulfur covalently bound to a
hydrogen or a carbon based group. Some non-limiting examples
include methylmercapto, ethylmercapto, n-propylmercapto,
isopropylmercapto or n-butylmercapto, ethylthio, n-propylthio or
isopropylthio group. The thio group also can be optionally
substituted with one or more moieties selected from the group
consisting of alkyl, halo, haloalkyl, hydroxyl, carboxyl, acyl,
acyloxy, amino, amido, carboxyl derivatives, alkylamino,
dialkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic
acid, thiol, imine, sulfonyl, sulfanyl, sulfinyl, sulfamonyl,
ester, carboxylic acid, amide, phosphonyl, phosphinyl, phosphoryl,
phosphine, thioester, thioether, acid halide, anhydride, oxime,
hydrozine, carbamate, phosphonic acid, phosphonate, or any other
viable functional group that does not inhibit the pharmacological
activity of this compound, either unprotected, or protected as
necessary, as known to those skilled in the art, for example, as
taught in Greene, et al., "Protective Groups in Organic Synthesis,"
John Wiley and Sons, 4.sup.th edition, 2007.
[0466] The term "ester" refers to a carbonyl flanked by an alkoxy
group and a carbon based group. Esters may be formed from the
reaction of a hydroxy group on the compound of the invention with a
carboxyl group of another group. Alternatively, an ester may be
formed by reaction of a carboxyl group on the compound of the
invention with a hydroxy group of another molecule. Some
non-limiting examples include hydroxycarbonyl, methoxycarbonyl,
ethoxycarbonyl, n-propyloxycarbonyl, isopropyloxycarbonyl,
n-butyloxycarbonyl, isobutyloxycarbonyl, tert-butyloxycarbonyl or
1-(cinnamyloxycarbonyloxy)-ethoxy-carbonyl. Esters of amino acids,
as used herein, include groups where a carboxyl group of the amino
acid forms an ester bond with a hydroxyl group of the molecule.
Also included are groups where a hydroxyl group on the amino acid
forms a ester bond with a carboxyl group on the molecule. The ester
group also can be optionally substituted with one or more moieties
selected from the group consisting of alkyl, halo, haloalkyl,
hydroxyl, carboxyl, acyl, acyloxy, amino, amido, carboxyl
derivatives, alkylamino, dialkylamino, arylamino, alkoxy, aryloxy,
nitro, cyano, sulfonic acid, thiol, imine, sulfonyl, sulfanyl,
sulfinyl, sulfamonyl, ester, carboxylic acid, amide, phosphonyl,
phosphinyl, phosphoryl, phosphine, thioester, thioether, acid
halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid,
phosphonate, or any other viable functional group that does not
inhibit the pharmacological activity of this compound, either
unprotected, or protected as necessary, as known to those skilled
in the art, for example, as taught in Greene, et al., "Protective
Groups in Organic Synthesis," John Wiley and Sons, 4.sup.th
edition, 2007.
III. Pharmaceutically Acceptable Salt formulations
[0467] Modifications of the active compound can affect the
bioavailability and rate of metabolism of the active species, thus
providing control over the delivery of the active species. Further,
the modifications can affect the activity of the compound, in some
cases increasing the activity over the parent compound. This can
easily be assessed by preparing the derivative and testing its
activity according to the methods described herein, or other method
known to those skilled in the art.
[0468] In cases where compounds are sufficiently basic or acidic to
form stable nontoxic acid or base salts, administration of the
compound as a pharmaceutically acceptable salt may be appropriate.
The term "pharmaceutically acceptable salts" or "complexes" refers
to salts or complexes that retain the desired biological activity
of the compounds of the present invention and exhibit minimal
undesired toxicological effects.
[0469] Examples of pharmaceutically acceptable salts are organic
acid addition salts formed with acids, which form a physiological
acceptable anion, for example, tosylate, methanesulfonate, acetate,
citrate, malonate, tartarate, succinate, benzoate, ascorbate,
.alpha.-ketoglutarate and .alpha.-glycerophosphate. Suitable
inorganic salts may also be formed, including, hydrochloride,
sulfate, nitrate, bicarbonate and carbonate salts. Alternatively,
the pharmaceutically acceptable salts may be made with sufficiently
basic compounds such as an amine with a suitable acid affording a
physiologically acceptable anion. Alkali metal (for example,
sodium, potassium or lithium) or alkaline earth metal (for example
calcium) salts of carboxylic acids can also be made.
[0470] Nonlimiting examples of such salts are (a) acid addition
salts formed with inorganic acids (for example, hydrochloric acid,
hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and
the like), and salts formed with organic acids such as acetic acid,
oxalic acid, tartaric acid, succinic acid, malic acid, ascorbic
acid, benzoic acid, tannic acid, pamoic acid, alginic acid,
polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic
acid, and polygalcturonic acid; (b) base addition salts formed with
metal cations such as zinc, calcium, bismuth, barium, magnesium,
aluminum, copper, cobalt, nickel, cadmium, sodium, potassium, and
the like, or with a cation formed from ammonia,
N,N-dibenzylethylenediamine, D-glucosamine, tetraethylammonium, or
ethylenediamine; or (c) combinations of (a) and (b); e.g., a zinc
tannate salt or the like. Also included in this definition are
pharmaceutically acceptable quaternary salts known by those skilled
in the art, which specifically include the quaternary ammonium salt
of the Formula --NR.sup.+A.sup.-, wherein R is as defined above and
A is a counterion, including chloride, bromide, iodide, --O-alkyl,
toluenesulfonate, methylsulfonate, sulfonate, phosphate, or
carboxylate (such as benzoate, succinate, acetate, glycolate,
maleate, malate, citrate, tartrate, ascorbate, benzoate,
cinnamoate, mandeloate, benzyloate, and diphenylacetate).
[0471] Pharmaceutically acceptable prodrugs refer to a compound
that is metabolized, for example hydrolyzed or oxidized, in the
host to form the compound of the present invention. Typical
examples of prodrugs include compounds that have biologically
labile protecting groups on a functional moiety of the active
compound. Prodrugs include compounds that can be oxidized, reduced,
aminated, deaminated, hydroxylated, dehydroxylated, hydrolyzed,
dehydrolyzed, alkylated, dealkylated, acylated, deacylated,
phosphorylated, dephosphorylated to produce the active
compound.
[0472] Any of the compounds described herein can be administered as
a prodrug to increase the activity, bioavailability, stability or
otherwise alter the properties of the compound. A number of prodrug
ligands are known. In general, alkylation, acylation or other
lipophilic modification of the compound will increase the stability
of the compound. Examples of substituent groups that can replace
one or more hydrogens on the compound are alkyl, aryl, steroids,
carbohydrates, including sugars, 1,2-diacylglycerol and alcohols.
Any of these can be used in combination with the disclosed
compounds to achieve a desired effect.
IV. Treatment of CNS Damage
[0473] The present invention provides methods and compositions for
the treatment or prevention of neurodegeneration following an
injury to the central nervous system or due to certain
neurodegenerative disorders comprising administering an effective
amount of steroid analog, or a pharmaceutically acceptable salt,
ester or prodrug thereof to a host in need thereof. Multiple
physiological events lead to neurodegeneration. These events
include, for example, increase in the immune and inflammatory
response, demyelinization, and lipid peroxidation. The present
invention provides compositions and methods for reducing or
eliminating neuronal cell death, edema, ischemia, and enhancing
tissue viability following injury to the central nervous system or
certain disorders. The analogues, salts, esters or prodrugs of the
steroid or secosteroid analogs may be optionally administered with
a pharmaceutically acceptable carrier or diluent.
[0474] By "treatment or prevention" is intended any enhanced
survival, proliferation, and/or neurite outgrowth of the neurons
that either prevents or retards neurodegeneration, the progressive
loss of neurons. As used herein, "neuroprotection" is the
prevention, arrest or reverse progression of neurodegeneration
following a central nervous system injury. The neuroprotective
effect includes both improved morphological (i.e., enhanced tissue
viability) and/or behavioral recovery. CNS injuries that are
encompassed within the scope of treatment of the present invention
include both traumatic injuries, in particular TBI, and
physiological insults such as an ischemic or hemorrhagic stroke. In
both instances, a progressive loss of neurons after the initial
insult occurs and can be alleviated by use of the inventive
compounds.
[0475] In certain embodiments, a method of preventing or reducing
inflammatory reactions in a patient is provided that includes
administering a steroid analog to a host in need thereof. In
certain embodiments, methods of neuroprotection are provided
comprising administering a compound of the invention, its
physiologically acceptable salt or prodrug, optionally in a
pharmaceutically acceptable carrier, to a patient at risk of
suffering from a stroke. In other embodiments, methods of treating
or preventing neuronal damage are provided comprising administering
a compound of the invention or its physiologically acceptable salt
or prodrug, optionally in a pharmaceutically acceptable carrier, to
a patient who has suffered from an ischemic stroke. The method can
reduce or prevent neurodegeneration such as that caused by
excitotoxic or inflammatory reactions, or can enhance neuronal
proliferation, growth or differentiation in the period after the
injury. In yet further embodiments, methods of treating or
preventing cognitive or behavioral deficits after a stroke is
provided comprising administering a compound of the invention or
its physiologically acceptable salt or prodrug, optionally in a
pharmaceutically acceptable carrier, to a patient who has suffered
a stroke. In certain embodiments, the stroke is an ischemic stroke,
but it can alternatively be a hemorrhagic stroke.
[0476] In other embodiments, the present invention provides a
method to achieve a neuroprotective effect following a traumatic
CNS injury in a mammal, in particular in a human, comprising
administering a therapeutically effective amount of steroid
analogue of the invention. A traumatic injury to the CNS is
characterized by a physical impact to the central nervous system.
The physical forces resulting in a traumatic brain injury cause
their effects by inducing three types of injury: skull fracture,
parenchymal injury, and vascular injury. A blow to the surface of
the brain typically leads to rapid tissue displacement, disruption
of vascular channels, and subsequent hemorrhage, tissue injury and
edema. Morphological evidence of injury in the neuronal cell body
includes pyknosis of nucleus, eosinophilia of the cytoplasm, and
disintegration of the cell. Furthermore, axonal swelling can
develop in the vicinity of damage neurons and also at great
distances away from the site of impact.
[0477] In certain embodiments, the compound is administered within
twelve hours after onset of a stroke. In certain embodiments, the
steroid analogue of the invention is administered within twelve
hours after an injury, such as a TBI. In some embodiments, the
compounds are administered within 11 hours of a TBI, stroke or
other injury to the brain, or within 10 hours, or within 9 hours,
or within 8 hours, or within 7 hours, or within 6 hours, or within
5 hours, or within 4 hours, or within 3 hours, such as within two
or one hour. In some other embodiments, the compounds are
administered within one day (i.e. 24 hours) of the injury. In
certain embodiments, the compounds are provided to individuals at
risk of a stroke, such as those who are suffering from
atherosclerosis or have a family history of heart disease. These
compounds can be provided to individuals as a preventative therapy
to decrease neural trauma.
[0478] In another embodiment, a method for decreasing ischemia
following a brain injury is provided comprising administering an
effective amount of a steroid analogue of the invention. The
methods of the invention provide a means to reduce or eliminate the
inflammatory immune reactions that follow a CNS injury. By reducing
the inflammatory response, the steroid analogues of the present
invention can substantially reduce brain swelling and reduce the
amount of neurotoxic substances (e.g., free radicals and
excitotoxins) that are released from the site of injury.
[0479] The present invention provides for a method of treating a
brain injury by administering to a subject steroid analogue of the
invention, a pharmaceutically acceptable salt or a prodrug or ester
thereof. The concentration of the steroid analogue or salt, ester
or prodrug thereof, in accordance with the present invention is
effective in the treatment or prevention of neuronal damage that
follows either a traumatic, ischemic or hemorrhagic injury to the
CNS and hence, elicits a neuroprotective effect. The
therapeutically effective amount will depend on many factors
including, for example, the specific activity of the steroid
analogue administered, the type of injury, the severity and pattern
of the injury, the resulting neuronal damage, the responsiveness of
the patient, the weight of the patient along with other intraperson
variability, the method of administration, and the formulation
used.
[0480] It is recognized that a traumatic injury to the CNS results
in multiple physiological events that impact the extent and rate of
neurodegeneration, and thus the final clinical outcome of the
injury. The treatment of a traumatic injury to the CNS, as defined
by the present invention, encompasses any reduction and/or
prevention in one or more of the various physiological events that
follow the initial impact. For example, cerebral edema frequently
develops following a traumatic injury to the CNS and is a leading
cause of death and disability. Cortical contusions, for example,
produce massive increases in brain tissue water content which, in
turn, can cause increased intracranial pressure leading to reduced
cerebral blood flow and additional neuronal loss. Hence, the
methods of the invention find use in reducing and/or eliminating
cerebral edema and/or reducing the duration of the edemic event
following a traumatic injury to the CNS. Assays to determine a
reduction in edema are known in the art and include, but are not
limited to, a decrease in tissue water content following the
administration of the progestin or the progestin metabolite (Betz
et al. (1990) Stroke 21:1199-204, which is herein incorporated by
reference). Furthermore, an overall improvement in behavioral
recovery can also be used as a measure for a decrease in edema. A
decrease in edema in the effected tissue by at least about 15% to
30%, about 30% to 45%, about 45% to 60%, about 60% to 80%, or about
80% to 95% or greater will be therapeutically beneficial, as will
any reduction in the duration of the edemic event.
[0481] Further physiological effects of brain injury include an
inflammatory response. In particular, some studies indicate that
the acute inflammatory response contributes significantly to injury
after ischemia (see Perera, et al. (2005) Inflammation following
stroke. J. Clin. Neurosc. 13:1-8; Barone and Feuerstein (1999)
Inflammatory mediators and stroke: new opportunities for novel
therapeutics). The stroke process triggers an inflammatory reaction
that may last up to several months. Suppression of inflammation can
reduce infarct volume and improve clinical outcomes even with the
initiation of therapy after 3 hours of onset of stroke. In
addition, an immune response can be triggered both by strokes.
Infiltrating leukocytes are thought to contribute to secondary
ischemic damage by producing toxic substances that kill brain cells
and disrupt the blood-brain barrier (see del Zoppo, et al. (2000)
Advances in the vascular pathophysiology of ischemic stroke. Thromb
Res. 98: 73-81)
Infiltration occurs when leukocytes bind endothelial intercellular
adhesion molecule-1 (ICAM-1) and ICAM-1 is upregulated after
ischemia.
[0482] TBI also elicits inflammatory, and in particular a immune
responses. See, for example, Soares et al. (1995) J. Neurosci.
15:8223-33; Holmin et al. (1995) Acta Neurochir. 132:110-9; Arvin
et al. (1996) Neurosci. Biobehav. Rev. 20:445-52. Following a
cortical impact, severe inflammatory reactions and gliosis at the
impact site and at brain areas distal to the primary site of injury
occurs. The inflammatory response is characterized by the
expression of adhesion molecules on the vascular surfaces,
resulting in the adherence of immune cells and subsequent
extravasation into the brain parenchyma. By releasing cytokines,
the invading macrophages and neutrophils stimulate reactive
astrocytosis. Release of different chemokines by other cell types
induces these immune cells to become phagocytic, with the
simultaneous release of free radicals and pro-inflammatory
compounds, e.g., cytokines, prostaglandins, and excitotoxins (Arvin
et al. (1996) Neurosci. Biobehav. Ref 20:445-52; Raivich et al.
(1996) Kelo J. Med. 45:239-47; Mattson et al. (1997) Brain Res.
Rev. 23:47-61; all of which are herein incorporated by
reference).
[0483] Assays for assessing the efficacy of the compounds described
herein include assays to determine a decrease in an ischemic event
include, for example, a decrease in infarct area, improved body
weight, and improved neurological outcome. Assays to measure a
reduction in lipid peroxidation in both brain homogenate and in
mitochondria are known in the art and include, for example, the
thiobarbituric acid method (Roof et al. (1997) Mol. Chem.
Neuropathol. 31: 1-11; Subramanian et al. (1993) Neurosci. Lett.
155:151-4; Goodman et al. (1996) J. Neurochem. 66:1836-44; Vedder
et al. (1999) J. Neurochem. 72:2531-8; all of which are herein
incorporated by reference) and various in vitro free radical
generating systems. Furthermore, alterations in the levels of
critical free radical scavenger enzymes, such as mitochondrial
glutathione can be assayed. See, for example, Subramanian et al.
(1993) Neurosci. Lett. 155:151-4; and Vedder et al. (1999) J.
Neurochem. 72:2531-8; both of which are herein incorporated by
reference.
[0484] Methods to quantify the extent of central nervous system
damage (i.e., neurodegeneration) and to determine if neuronal
damage was treated or prevented following the administration of a
progesterone or steroid analogue are well known in the art. Such
neuroprotective effects can be assayed at various levels,
including, for example, by promoting behavioral and morphological
(i.e., enhancing tissue viability) recovery after traumatic brain
injury. A variety of anatomical, immunocytochemical and
immunological assays to determine the effect of the progestin
metabolite on necrosis, apoptosis, and neuronal glial repair are
known in the art. As such, the neuroprotection resulting from the
methods of the present invention will result in at least about a
10% to 20%, 20% to 30%, 30% to 40%, 40% to 60%, 60% to 80% or
greater increase in neuronal survival and/or behavioral recovery as
compared to the control groups.
[0485] Histological and molecular marker assays for an increase in
neuronal survival are known. For example, Growth Associated Protein
43 (GAP-43) can be used as a marker for new axonal growth following
a CNS insult. See, for example, Stroemer et al. (1995) Stroke
26:2135-2144, Vaudano et al. (1995) J. of Neurosci 15:3594-3611.
Other histological markers can include a decrease in astrogliosis
and microgliosis. Alternatively, a delay in cellular death can be
assayed using TUNEL labeling in injured tissue. Further anatomical
measures that can be used to determine an increase in
neuroprotection include counting specific neuronal cell types to
determine if the progestin or the progestin metabolite is
preferentially preserving a particular cell type (e.g., cholinergic
cells) or neurons in general.
[0486] In addition, behavioral assays can be used to determine the
rate and extent of behavior recovery in response to the treatment.
Improved patient motor skills, spatial learning performance,
cognitive function, sensory perception, speech and/or a decrease in
the propensity to seizure may also be used to measure the
neuroprotective effect. Such functional/behavioral tests used to
assess sensorimortor and reflex function are described in, for
example, Bederson et al. (1986) Stroke 17:472-476, DeRyck et al.
(1992) Brain Res. 573:44-60, Markgraf et al. (1992) Brain Res.
575:238-246, Alexis et al. (1995) Stroke 26:2336-2346; all of which
are herein incorporated by reference. Enhancement of neuronal
survival may also be measured using the Scandinavian Stroke Scale
(SSS) or the Barthl Index. Behavioral recovery can be further
assessed using the recommendations of the Subcommittee of the
NIH/NINDS Head Injury Centers in Humans (Hannay et al. (1996) J.
Head Trauma Rehabil. 11:41-50), herein incorporated by reference.
Behavioral recovery can be further assessed using the methods
described in, for example, Beaumont et al. (1999) Neurol Res.
21:742-754; Becker et al. (1980) Brain Res. 200:07-320; Buresov et
al. (1983) Techniques and Basic Experiments for the Study of Brain
and Behavior; Kline et al. (1994) Pharmacol. Biochem. Behav.
48:773-779; Lindner et al. (1998) J. Neurotrauma 15:199-216; Morris
(1984) J. Neurosci. Methods 11:47-60; Schallert et al. (1983)
Pharmacol. Biochem. Behav. 18:753-759.
[0487] Assays that can be used to determine if the steroid analogue
of the invention is imparting an anti-inflammatory and a
nonspecific suppressive effect on the immune system following a
traumatic CNS injury include, for example, a reduction in cytokine
induced microglial proliferation in vitro (Hoffman et al. (1994) J.
Neurotrauma 11:417-31; Garcia-Estrada et al. (1993) Brain Res.
628:271-8; both of which are herein incorporated by reference); a
reduction in the generation of cytotoxic free radicals by activated
macrophages (Chao et al. (1994) Am. J. Reprod. Immunol. 32:43-52;
Robert et al. (1997) Nitric Oxide 1:453-62; Kelly et al. (1997)
Biochem. Biophys. Res. Commun. 239:557-61; Ganter et al. (1992) J.
Neurosci. Res. 33:218-30; all of which are herein incorporated by
reference); a reduction in the expression of inducible nitric oxide
synthetase and the amount of nitric oxide release by macrophages
(Robert et al. (1997) Nitric Oxide 1:453-62; Miller et al. (1996)
J. Leukoc. Biol. 59:442-50; both of which are herein incorporated
by reference); the release of a "progesterone-induced blocking
factor" that inhibits natural killer cell activity (Cheek et al.
(1997) Am. J. Reprod. Immunol. 37:17-20; Szekeres-Bartho et al.
(1997) Cell Immunol. 177:194-9; Szekeres-Bartho et al. (1996) Am.
J. Reprod. Immunol. 35:348-51; all of which are herein incorporated
by reference); a decrease in the number of GFAP-positive astrocytes
after brain injury which is suggestive of less secondary damage
(Garcia-Estrada et al. (1993) Brain Res. 628:271-8; Garcie-Estrada
et al. (1999) Int. J. Dev. Neurosci. 17:145-51; Cheek et al. (1997)
Am. J. Reprod. Immunol. 37:17-20; Szekeres-Bartho et al. (1997)
Cell Immunol. 177:194-9; Szekeres-Bartho et al. (1996) Am. J.
Reprod. Immunol. 35:348-51; all of which are herein incorporated by
reference); a reduction in the number of inflammatory immune cells
(OX42-positive cells); a reduction in the loss of ChAT-positive and
COX-positive neurons; a reduction in the number of TUNEL-positive
and MnSOD-positive neurons; and an increase in the intensity of
succinate dehydrogenase and cytochrome oxidase activity.
[0488] Furthermore, a reduction in the inflammatory immune
reactions following a traumatic brain injury can be assayed by
measuring the cytokines level following the injury in the sham
controls versus the progestin treated subjects. Cytokines are
mediators of inflammation and are released in high concentrations
after brain injury. The level of pro-inflammatory cytokines (e.g.,
interleukin 1-beta, tumor necrosis factor, and interleukin 6) and
the level of anti-inflammatory cytokines (e.g., interleukin 10 and
transforming growth factor-beta) can be measured. For instance,
"real-time" polymerase chain reactions (PCR) can be used to measure
the strength of the mRNA signal and ELISA can be used to determine
protein levels. In addition, histological analysis for different
inflammatory cell types (e.g., reactive astrocytes, macrophages and
microglia) can be used to measure a reduction in the inflammatory
response.
[0489] The compounds of the invention can also have potential for
use in other disorders including multiple sclerosis, catamenial
epilepsy, diabetic neuropathy, inflammatory disorders (e.g.,
rheumatoid arthritis, inflammatory bowel disease), hemorrhagic
shock, Niemann-Pick disorder, cerebral palsy, and congenital heart
disorders.
[0490] In specific embodiments, a method of treatment or prevention
of neural degeneration related to Amyotrophic Lateral Sclerosis
(ALS), is provided comprising administering a steroid analog
described herein, or a pharmaceutically acceptable salt, ester or
prodrug thereof, to a patient suffering from or at risk of
suffering from ALS. ALS, more commonly known as Lou Gehrig's
Disease, strikes both males and females, typically between the ages
of 40 and 70. This is a motor neuron disorder in which both the
upper and lower motor neurons are affected. Patients' muscles
atrophy as the motor neurons cease sending signals to initiate
movement. This affects not only muscles required for locomotion but
also the muscles used in swallowing. Up until the age of 60, males
are disproportionally affected at a ratio of 1.5 to 1. After the
age of 60, the numbers are equal across genders. The incidence of
ALS is approximately 1/2 that of multiple sclerosis. Life
expectancy post-diagnosis is 2-5 years. There are 120,000 cases of
ALS diagnosed worldwide and 350,000 patients coping with the
disease at any given time. A treatment for ALS will clearly qualify
for orphan drug status. The cause of ALS has not been identified.
The pathogenesis is poorly understood but excitotoxicity,
inflammation, oxidative stress and protein aggregation have been
shown. In some cases, super oxide dismutase 1 (SOD 1) has been
determined to be aberrant. Glutamate toxicity is now generally
accepted as part of AS pathology. Progesterone has proven to
protect neurons from the effects of this toxicity. The only
compound approved for the treatment of ALS is Rilutek.TM. which may
reduce glutamate levels. It is not curative but has reduced the
rate of progression in some patients.
[0491] In another specific embodiments, a method of treatment or
prevention of neural degeneration related to Parkinson's Disease
(PD), is provided comprising administering a steroid analog
described herein, or a pharmaceutically acceptable salt, ester or
prodrug thereof, to a patient suffering from or at risk of
suffering from PD. PD is a neurodegenerative disease of unknown
etiology that results in the progressive loss of nerve cell
function in the brain. Life expectancy is 15-25 years
post-diagnosis; however, there is no cure. It is estimated that one
million people in the U.S. are living with Parkinson's; a number
that is greater than the combined total of multiple sclerosis,
muscular dystrophy and amyotrophic lateral sclerosis patients. The
incidence of PD increases with age. Nearly 40,000 people are
diagnosed each year with PD, of which .about.15% will be less than
50 years in age. The cost of PD annually exceeds $25 billion when
both direct and indirect costs are combined. In PD, cells in the
substania nigra of the brain cease to function properly and die.
These cells produce dopamine, a neurotransmitter. Dopamine
regulates those parts of the brain which control the initiation of
movement and coordination. Without dopamine, a patient will begin
to experience tremors, bradykinesia, postural instability, rigidity
of limbs and trunk, and/or impaired balance and coordination. Not
all patients experience all symptoms nor do they progress at the
same rate. PD is ultimately debilitating for many sufferers who
require assistance in everyday living.
[0492] In another specific embodiments, a method of treatment or
prevention of neural degeneration related to spinal cord trauma is
provided comprising administering a steroid analog described
herein, or a pharmaceutically acceptable salt, ester or prodrug
thereof, to a patient in need thereof. In another specific
embodiments, a method of treatment or prevention of neural
degeneration related to hypoxia is provided comprising
administering a steroid analog described herein, or a
pharmaceutically acceptable salt, ester or prodrug thereof, to a
patient in need thereof.
V. Combination and Alternation Therapy
[0493] In further embodiments of the present invention, the steroid
analogues of the invention may be administered in combination or
alternation with at least one additional neuroprotective agent to
enhance neuroprotection following a traumatic CNS injury. In one
embodiment, the inventive steroid analogues of the invention may be
administered in combination or alternation with other steroid
analogues or with progesterone.
[0494] Other neuroprotective agents of interest include, for
example, compounds that reduce glutamate excitotoxicity and enhance
neuronal regeneration. Such agents may be selected from, but not
limited to, the group comprising growth factors. As used herein,
"growth factor" refers to an extracellular polypeptide-signaling
molecule that stimulates a cell to grow or proliferate. Preferred
growth factors are those to which a broad range of cell types
respond. Examples of neurotrophic growth factors include, but are
no limited to, fibroblast growth factor family members such as
basic fibroblast growth factor (bFGF) (Abraham et al. (1986)
Science 233:545-48), acidic fibroblast growth factor (aFGF) (Jaye
et al. (1986) Science 233:541-45), the hst/Kfgf gene product, FGF-3
(Dickson et al. (1987) Nature 326-833), FGF-4 (Zhan et al. (1988)
Mol. Cell. Biol. 8:3487-3495), FGF-6 (deLapeyriere et al. (1990)
Oncogene 5:823-831), keratinocyte growth factor (KGF) (Finch et al.
(1989) Science 245:752-755), and androgen-induced growth factor
(AIGF) (Tanaka et al. (1992) Proc. Natl. Acad. Sci. USA
89:8928-8923).
[0495] Additional neuroprotective agents include, ciliary
neurotrophic factor (CNTF), nerve growth factor (NGF) (Seiler, M.
(1984) Brain Research 300:33-39; Hagg T. et al. (1988) Exp Neurol
101:303-312; Kromer L. F. (1987) Science 235:214-216; and Hagg T.
et al. (1990) J. Neurosci 10(9):3087-3092), brain derived
neurotrophic factor (BDNF) (Kiprianova, I. et al. (1999) J.
Neurosci. Res. 56:21-27), Neurotrophin 3 (NT3), Neurotrophin 4
(NT4), transforming growth factor-.beta.1 (TGF-.beta.1)
(Henrick-Noack, P. et al. (1996) Stroke 27:1609-14), bone
morphogenic protein (BMP-2) (Hattori, A. et al. (1999) J.
Neurochem. 72:2264-71), glial-cell line derived neurotrophic factor
(GDNF) (Miyazaki, H. et al. (1999) Neuroscience 89:643-7),
activity-dependant neurotrophic factor (ADNF) (Zamostiano, R. et
al. (1999) Neurosci Letter 264:9-12), cytokine leukemia inhibiting
factor (LIF) (Blesch, A. et al. (1999) J. Neurosci. 19:3356-66),
oncostatin M, interleukin, and the insulin-like growth factors 1
and 2.
[0496] Other forms of neuroprotective therapeutic agents include,
for example, Clomethiazole (Zendra) (Marshal, J. W. et al. (1999)
Exp. Neurol. 156:121-9); kynurenic acid (KYNA) (Salvati, P. et al.
(1999) Prog Neruopsychopharmacol Biol Psychiatry 23:741-52), Semax
(Miasoedova, N. F. et al. (1999) Zh Nevrol Psikhiatr Imss Korsakova
99:15-19), FK506 (tacrolimus) (Gold, B. G. et al. (1999) J.
Pharmacol. Exp. Ther. 289:1202-10),
L-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (Inokuchi,
J. et al. (1998) Act Biochim Pol 45:479-92),
andrenocorticotropin-(4-9) analoge (ORG 2766) and dizolcipine
(MK-801) (Herz, R. C. et al. (1998) Eur J. Pharmacol 346:159-65),
cerebral interleukin-6) (Loddick, S. A. et al. (1998) J. Cereb
Blood Flow Metab 18:176-9), selegiline (Semkova, I. et al. (1996)
Eur J. Pharmacol 315:19-30), MK-801 (Barth, A. et al. (1996) Neuro
Report 7:1461-4; glutamate antagonist such as, NPS1506, GV1505260,
MK801 (Baumgartner, W. A. et al. (1999) Ann Thorac Surg 67:1871-3),
GV150526 (Dyker, A. G. et al. (1999) Stroke 30:986-92); AMPA
antagonist such as NBQX (Baumgartner, W. A. (1999) et al. Ann
Thorac Surg 67:1871-3, PD152247 (PNQX) (Schielke, G. P. et al.
(1999) Stroke 30:1472-7), SPD 502 (Nielsen, E. O. et al. (1999) J.
Pharmacol Exp Ther 289:1492-501), LY303070 and LY300164 (May, P. C.
et al. (1999) Neuroscience Lett 262:219-221).
[0497] When the steroid analogues of the present invention are
administered in combination or alternation with other
pharmaceutically active agents, (i.e., other neuroprotective
agents) even less of the steroid analogue may be therapeutically
effective. The steroid analogue may be administered once or several
times a day. The duration of the treatment may be once per day for
a period of from two to three weeks and may continue for a period
of months or even years. The daily dose can be administered either
by a single dose in the form of an individual dosage unit or
several smaller dosage units or by multiple administration of
subdivided dosages at certain intervals.
[0498] For example, a dosage unit can be administered from 0 hours
to 1 hr, 1 hr to 24 hr or 24 hours to at least 100 hours post
injury. Alternatively, the dosage unit can be administered from
about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 30, 40, 48, 72, 96, 120 hours or
longer post injury. Subsequent dosage units can be administered any
time following the initial administration such that a therapeutic
effect is achieved. For instance, additional dosage units can be
administered to protect the subject from the secondary wave of
edema that may occur over the first several days post-injury.
[0499] In combination therapy, effective dosages of two or more
agents are administered together, whereas during alternation
therapy an effective dosage of each agent is administered serially.
The dosages will depend on absorption, inactivation and excretion
rates of the drug as well as other factors known to those of skill
in the art. It Dosage values will also vary with the severity of
the condition to be alleviated. For any particular subject,
specific dosage regimens and schedules should be adjusted over time
according to the individual need and the professional judgment of
the person administering or supervising the administration of the
compositions.
[0500] When the compounds or compositions of the present invention
are administered in combination or alternation with other
pharmaceutically active agents, (i.e., other neuroprotective
agents) a lower level of the steroid analog in certain embodiments
may be used. The compositions may be administered once or several
times a day. The duration of the treatment may be once per day for
a period of from two to three weeks and may continue for a period
of months or even years. The daily dose can be administered either
by a single dose in the form of an individual dosage unit or
several smaller dosage units or by multiple administration of
subdivided dosages at certain intervals.
[0501] For example, a dosage unit can be administered from 0 hours
to 1 hr, 1 hr to 24 hr or 24 hours to at least 100 hours post
injury. Alternatively, the dosage unit can be administered from
about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,
17, 18, 19, 20, 21, 22, 23, 24, 30, 40, 48, 72, 96, 120 hours or
longer post injury. Subsequent dosage units can be administered any
time following the initial administration such that a therapeutic
effect is achieved. For instance, additional dosage units can be
administered to protect the subject from the secondary wave of
edema that may occur over the first several days post-injury.
[0502] In combination therapy, effective dosages of two or more
agents are administered together, whereas during alternation
therapy an effective dosage of each agent is administered serially.
The dosages will depend on absorption, inactivation and excretion
rates of the drug as well as other factors known to those of skill
in the art. It is to be noted that dosage values will also vary
with the severity of the condition to be alleviated. It is to be
further understood that for any particular subject, specific dosage
regimens and schedules should be adjusted over time according to
the individual need and the professional judgment of the person
administering or supervising the administration of the
compositions.
[0503] The efficacy of a drug can be prolonged, augmented, or
restored by administering the compound in combination or
alternation with a second, and perhaps third, agent. Alternatively,
the pharmacokinetics, biodistribution or other parameter of the
drug can be altered by such combination or alternation therapy. In
general, combination therapy is typically preferred over
alternation therapy because it induces multiple simultaneous
stresses on the condition.
[0504] In another embodiment, the active compound is administered
in combination or alternation with one or more other non-steroidal
anti-inflammatory drug(s) (NSAIDS). Examples ofNSAIDS that can be
used in alternation or combination therapy are carboxylic acids,
propionic acids, fenamates, acetic acids, pyrazolones, oxicans,
alkanones, gold compounds and others that inhibit prostaglandin
synthesis, preferably by selectively inhibiting cylcooxygenase-2
(COX-2). Some nonlimiting examples of COX-2 inhibitors are Celebrex
(celecoxib) and Vioxx (rofacoxib). Some non-limiting examples
ofNSAIDS are aspirin (acetylsalicylic acid), Dolobid (diflunisal),
Disalcid (salsalate, salicylsalicylate), Trisilate (choline
magnesium trisalicylate), sodium salicylate, Cuprimine
(penicillamine), Tolectin (tolmetin), ibuprofen (Motrin, Advil,
Nuprin Rufen), Naprosyn (naproxen, Anaprox, naproxen sodium),
Nalfon (fenoprofen), Orudis (ketoprofen), Ansaid (flurbiprofen),
Daypro (oxaprozin), meclofenamate (meclofanamic acid, Meclomen),
mefenamic acid, Indocin (indomethacin), Clinoril (sulindac),
tolmetin, Voltaren (diclofenac), Lodine (etodolac), ketorolac,
Butazolidin (phenylbutazone), Tandearil (oxyphenbutazone),
piroxicam (Feldene), Relafen (nabumetone), Myochrysine (gold sodium
thiomalate), Ridaura (auranofin), Solganal (aurothioglucose),
acetaminophen, colchicine, Zyloprim (allopurinol), Benemid
(probenecid), Anturane (sufinpyrizone), Plaquenil
(hydroxychloroquine), Aceclofenac, Acemetacin, Acetanilide,
Actarit, Alclofenac, Alminoprofen, Aloxiprin, Aluminium Aspirin,
Amfenac Sodium, Amidopyrine, Aminopropylone, Ammonium Salicylate,
Ampiroxicam, Amyl Salicylate, Anirolac, Aspirin, Auranofin,
Aurothioglucose, Aurotioprol, Azapropazone, Bendazac (Bendazac
Lysine), Benorylate, Benoxaprofen, Benzpiperylone, Benzydamine
hydrochloride, Bomyl Salicylate, Bromfenac Sodium, Bufexamac,
Bumadizone Calcium, Butibufen Sodium, Capsaicin, Carbaspirin
Calcium, Carprofen, Chlorthenoxazin, Choline Magnesium
Trisalicylate, Choline Salicylate, Cinmetacin, Clofexamide,
Clofezone, Clometacin, Clonixin, Cloracetadol, Cymene, Diacerein,
Diclofenac (Diclofenac Diethylammonium Salt, Diclofenac Potassium,
Diclofenac Sodium), Diethylamine Salicylate, Diethylsalicylamide,
Difenpiramide, Diflunisal, Dipyrone, Droxicam, Epirizole,
Etenzamide, Etersalate, Ethyl Salicylate, Etodolac, Etofenamate,
Felbinac, Fenbufen, Fenclofenac, Fenoprofen Calcium, Fentiazac,
Fepradinol, Feprazone, Floctafenine, Flufenamic, Flunoxaprofen,
Flurbiprofen (Flurbiprofen Sodium), Fosfosal, Furprofen, Glafenine,
Glucametacin, Glycol Salicylate, Gold Keratinate, Harpagophytum
Procumbens, Ibufenac, Ibuprofen, Ibuproxam, Imidazole Salicylate,
Indomethacin (Indomethacin Sodium), Indoprofen, Isamifazone,
Isonixin, Isoxicam, Kebuzone, Ketoprofen, Ketorolac Trometamol,
Lithium Salicylate, Lonazolac Calcium, Lomoxicam, Loxoprofen
Sodium, Lysine Aspirin, Magnesium Salicylate, Meclofenamae Sodium,
Mefenamic Acid, Meloxicam, Methyl Butetisalicylate, Methyl
Gentisate, Methyl Salicylate, Metiazinic Acid, Metifenazone,
Mofebutazone, Mofezolac, Morazone Hydrochloride, Morniflumate,
Morpholine Salicylate, Nabumetone, Naproxen (Naproxen Sodium),
Nifenazone, Niflumic Acid, Nimesulide, Oxametacin, Oxaprozin,
Oxindanac, Oxyphenbutazone, Parsalmide, Phenybutazone, Phenyramidol
Hydrochloride, Picenadol Hydrochloride, Picolamine Salicylate,
Piketoprofen, Pirazolac, Piroxicam, Pirprofen, Pranoprofen,
Pranosal, Proglumetacin Maleate, Proquazone, Protizinic Acid,
Ramifenazone, Salacetamide, Salamidacetic Acid, Salicylamide,
Salix, Salol, Salsalate, Sodium Aurothiomalate, Sodium Gentisate,
Sodium Salicylate, Sodium Thiosalicylate, Sulindac, Superoxide
Dismutase (Orgotein, Pegorgotein, Sudismase), Suprofen, Suxibuzone,
Tenidap Sodium, Tenoxicam, Tetrydamine, Thurfyl Salicylate,
Tiaprofenic, Tiaramide Hydrochloride, Tinoridine Hydrochloride,
Tolfenamic Acid, Tometin Sodium, Triethanolamine Salicylate,
Ufenamate, Zaltoprofen, Zidometacin and Zomepirac Sodium.
VI. Pharmaceutical Compositions
[0505] The described compounds can be formulated as pharmaceutical
compositions and administered for the treatment or prevention of
CNS injury from trauma or disease, and in particular for injury
resulting from traumatic brain injury or stroke. The compositions
can be administered in any of a variety of forms adapted to the
chosen route of administration, including systemically, such as
orally, or parenterally, by intravenous, intramuscular, topical,
transdermal or subcutaneous routes.
[0506] The compound can be included in the pharmaceutically
acceptable carrier or diluent in an amount sufficient to deliver to
a patient a therapeutically effective amount of compound to treat
traumatic CNS injury in vivo without causing serious toxic effects
in the patient treated.
[0507] The steroid analogues used in the methods of the invention
may further comprise an inorganic or organic, solid or liquid,
pharmaceutically acceptable carrier. The carrier may also contain
preservatives, wetting agents, emulsifiers, solubilizing agents,
stabilizing agents, buffers, solvents and salts. Compositions may
be sterilized and exist as solids, particulants or powders,
solutions, suspensions or emulsions.
[0508] The steroid analogues can be formulated according to known
methods to prepare pharmaceutically useful compositions, such as by
admixture with a pharmaceutically acceptable carrier vehicle.
Suitable vehicles and their formulation are described, for example,
in Remington: The Science and Practice of Pharmacy, 21st ed,
Lippincott Williams & Wilkins (2005). In order to form a
pharmaceutically acceptable composition suitable for effective
administration, such compositions will contain an effective amount
of the steroid analogue, either alone, or with a suitable amount of
carrier vehicle.
[0509] The pharmaceutically acceptable carrier of the present
invention will vary depending on the method of drug administration.
The pharmaceutical carrier employed may be, for example, either a
solid, liquid, or time release. Representative solid carriers are
lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia,
magnesium stearate, stearic acid, microcrystalin cellulose, polymer
hydrogels, and the like. Typical liquid carriers include syrup,
peanut oil, olive oil, cyclodextrin, and the like emulsions. Those
skilled in the art are familiar with appropriate carriers for each
of the commonly utilized methods of administration. Furthermore, it
is recognized that the total amount of steroid analogue
administered as a therapeutic effective dose will depend on both
the pharmaceutical composition being administered (i.e., the
carrier being used) and the mode of administration.
[0510] In one embodiment, a steroid analogue, pharmaceutically
acceptable salt, ester or prodrug thereof, is administered via
parenteral administration in a dose of about 0.1 ng to about 100 g
per kg of body weight, about 10 ng to about 50 g per kg of body
weight, from about 100 ng to about 1 g per kg of body weight, from
about 1 .mu.g to about 100 mg per kg of body weight, from about 1
.mu.g to about 50 mg per kg of body weight, from about 1 mg to
about 500 mg per kg of body weight; and from about 1 mg to about 50
mg per kg of body weight. Alternatively, the amount of steroid
analogue administered to achieve a therapeutic effective dose is
about 0.1 ng, 1 ng, 10 ng, 100 ng, 1 .mu.g, 10 .mu.g, 100 .mu.g, 1
mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg,
12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 30
mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 500 mg per kg
of body weight or greater.
[0511] In certain embodiments, the compounds described herein are
compounded with a suitable pharmaceutically acceptable carrier in a
unit dosage form. A unit dosage form, such as a preselected amount
of liquid composition, can, for example, contain the compound in
amounts ranging from about 5 to about 1000 mg, or from about 250 to
about 750 mg. Expressed in proportions, the active compound is
generally present in from about 10 to about 750 mg/ml of carrier.
Liquid formulations of steroid analogs can comprise about 1-100
mg/ml of vehicle. In the case of compositions containing
supplementary active ingredients, the dosages are determined by
reference to the usual dose and manner of administration of the
said ingredients.
[0512] The active ingredients can exhibit activity, particularly in
treatment or prevention of secondary reactions from brain injuries
such as TBI or stroke when administered in amounts ranging from
about 0.1 mg to about 100 mg per kilogram of body weight per day. A
preferred dosage regimen for optimum results would be from about
0.5 mg to about 50 mg per kilogram of body weight per day, and such
dosage units are employed that a total of from about 0.25 gram to
about 3.0 grams of the active compound for a subject of about 70 kg
of body weight are administered in a 24-hour period. This dosage
regimen may be adjusted to provide the optimum therapeutic response
and can be administered one to three times a day in dosages of
about 600 mg per administration. For example, several divided doses
may be administered daily or the dose may be proportionally reduced
as indicated by the exigencies of the therapeutic situation.
[0513] In one embodiment of the present invention, the
neuroprotective steroid is administered once or several times a
day. The duration of the treatment may be once per day for a period
of about 1, 2, 3, 4, 5, 6, 7 days or more. The daily dose can be
administered either by a single dose in the form of an individual
dosage unit or several smaller dosage units or by multiple
administration of subdivided dosages at certain intervals. For
instance, a dosage unit can be administered from about 0 hours to
about 1 hr, about 1 hr to about 24 hr, about 1 to about 72 hours,
about 1 to about 120 hours, or about 24 hours to at least about 120
hours post injury. Alternatively, the dosage unit can be
administered from about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 30, 40, 48, 72, 96,
120 hours or longer post injury. The duration of the constant
dosing regimen is about 12, 24, 36, 60, 72, 84, or 120 hours or
about 1 to 24 hours, about 12 to 36 hours, about 24 to 48 hours,
about 36 to 60 hours, about 48 to 72 hours, about 60 to 96 hours,
about 72 to 108 hours, about 96 to 120 hours, or about 108 to 136
hours. Subsequent dosage units can be administered any time
following the initial administration such that a therapeutic effect
is achieved. For instance, additional dosage units can be
administered to protect the subject from the secondary wave of
edema that may occur over the first several days post-injury. In
specific embodiments, the subject undergoing the therapy with is
administered a constant neuroprotective steroid dosing regimen. By
"constant dosing regimen" is intended the subject undergoing
therapy is administered a constant total hourly infusion dose over
the course of treatment.
[0514] Administration of the steroid analogues of the invention may
be performed by many methods known in the art. The present
invention comprises all forms of dose administration including, but
not limited to, systemic injection, parenteral administration,
intravenous, intraperitoneal, intramuscular, transdermal, buccal,
subcutaneous and intracerebroventricular administration.
Alternatively, the steroid analogue may be administered directly
into the brain or cerebrospinal fluid by any
intracerebroventricular technique including, for example, lateral
cerebro ventricular injection, lumbar puncture or a surgically
inserted shunt into the cerebro ventricle of a patient. Methods of
administering may be by dose or by control release vehicles.
[0515] If administered intravenously, preferred carriers are
physiological saline or phosphate buffered saline (PBS).
[0516] While the methods of the invention are not bound by any
theory, it is believed that a traumatic CNS injury, may make the
blood/brain barrier more permeable allowing entry of large
molecules that would not normally cross the blood/brain barrier to
enter the cerebral spinal fluid. For examples of intravenous,
intraperitoneal, intramuscular, and subcutaneous administration of
neurotrophic agents to treat CNS injuries see, for example, U.S.
Pat. No. 5,733,871 and WO 97/21449, both of which are herein
incorporated by reference.
[0517] Additional pharmaceutical methods may be employed to control
the duration of action. Controlled release preparations may be
achieved by the use of polymers to complex or absorb the steroid
analogue. The controlled delivery may be exercised by selecting
appropriate macromolecules (for example, polyesters, polyamino
acids, polyvinyl pyrrolidone, ethylene-vinylacetate,
methylcellulose, carboxymethylcellulose, or protamine sulfate). The
rate of drug release may also be controlled by altering the
concentration of such macromolecules.
[0518] Another possible method for controlling the duration of
action comprises incorporating the therapeutic agents into
particles of a polymeric substance such as polyesters, polyamino
acids, hydrogels, poly(lactic acid) or ethylene vinylacetate
copolymers. Alternatively, it is possible to entrap the therapeutic
agents in microcapsules prepared, for example, by coacervation
techniques or by interfacial polymerization, for example, by the
use of hydroxymethyl cellulose or gelatin-microcapsules or
poly(methylmethacrylate) microcapsules, respectively, or in a
colloid drug delivery system, for example, liposomes, albumin,
microspheres, microemulsions, nanoparticles, nanocapsules, or in
macroemulsions. Such teachings are disclosed in Remington's
Pharmaceutical Sciences (1980). Ideally the compounds should be
administered to achieve peak plasma concentrations of the active
compound of from about 0.2 to 70 .mu.M, preferably about 1.0 to 10
.mu.M. This may be achieved, for example, by the intravenous
injection of an appropriate concentration of the active ingredient,
optionally in saline, or administered as a bolus of the active
ingredient.
[0519] The concentration of the compounds in the drug composition
will depend on absorption, inactivation and excretion rates of the
extract as well as other factors known to those of skill in the
art. It is to be noted that dosage values will also vary with the
severity of the condition to be alleviated. It is to be further
understood that for any particular subject, specific dosage
regimens should be adjusted over time according to the individual
need and the professional judgment of the person administering or
supervising the administration of the compositions, and that the
concentration ranges set forth herein are exemplary only and are
not intended to limit the scope or practice of the claimed
composition. The compounds may be administered at once, or may be
divided into a number of smaller doses to be administered at
varying intervals of time.
[0520] Oral compositions will generally include an inert diluent or
an edible carrier. They may be enclosed in gelatin capsules or
compressed into tablets. For the purpose of oral therapeutic
administration, the active compound can be incorporated with
excipients and used in the form of tablets, troches or capsules.
Pharmaceutically compatible binding agents, and/or adjuvant
materials can be included as part of the composition.
[0521] The tablets, pills, capsules, troches and the like can
contain any of the following ingredients, or compounds of a similar
nature: a binder such as microcrystalline cellulose, gum tragacanth
or gelatin; an excipient such as starch or lactose, a
disintegrating agent such as alginic acid, Primogel, or corn
starch; a lubricant such as magnesium stearate or Sterotes; a
glidant such as colloidal silicon dioxide; a sweetening agent such
as sucrose or saccharin; or a flavoring agent such as peppermint,
methyl salicylate, or orange flavoring. When the dosage unit form
is a capsule, it can contain, in addition to material of the above
type, a liquid carrier such as a fatty oil. In addition, dosage
unit forms can contain various other materials which modify the
physical form of the dosage unit, for example, coatings of sugar,
shellac, or other enteric agents.
[0522] The compounds can be administered as a component of an
elixir, suspension, syrup, wafer, chewing gum or the like. A syrup
may contain, in addition to the active compounds, sucrose as a
sweetening agent and certain preservatives, dyes and colorings and
flavors. The compounds can also be mixed with other active
materials that do not impair the desired action, or with materials
that supplement the desired action, such as antibiotics,
antifungals, anti-inflammatories, or other anti-autoimmune
compounds. Solutions or suspensions used for parenteral,
intradermal, subcutaneous, or topical application can include the
following components: a sterile diluent such as water for
injection, saline solution, fixed oils, polyethylene glycols,
glycerine, propylene glycol or other synthetic solvents;
antibacterial agents such as benzyl alcohol or methyl parabens;
antioxidants such as ascorbic acid or sodium bisulfite; chelating
agents such as ethylenediaminetetraacetic acid; buffers such as
acetates, citrates or phosphates and agents for the adjustment of
tonicity such as sodium chloride or dextrose. The parental
preparation can be enclosed in ampoules, disposable syringes or
multiple dose vials made of glass or plastic.
[0523] Formulations suitable for parental administration
conveniently comprise a sterile aqueous preparation of the active
compound, which can be isotonic with the blood of the
recipient.
[0524] Nasal spray formulations comprise purified aqueous solutions
of the active agent with preservative agents and isotonic agents.
Such formulations are preferably adjusted to a pH and isotonic
state compatible with the nasal mucous membranes.
[0525] Formulations for rectal administration may be presented as a
suppository with a suitable carrier such as cocoa butter, or
hydrogenated fats or hydrogenated fatty carboxylic acids.
[0526] Ophthalmic formulations are prepared by a similar method to
the nasal spray, except that the pH and isotonic factors are
preferably adjusted to match that of the eye.
[0527] Topical formulations comprise the active compound dissolved
or suspended in one or more media such as mineral oil, petroleum,
polyhydroxy alcohols or other bases used for topical formulations.
The addition of other accessory ingredients as noted above may be
desirable.
[0528] Further, the present invention provides liposomal
formulations of the steroid analogues, salts, esters and prodrugs
thereof. The technology for forming liposomal suspensions is well
known in the art. When the steroid analogue or salt thereof is an
aqueous-soluble salt, using conventional liposome technology, the
same may be incorporated into lipid vesicles. In such an instance,
due to the water solubility of the compound or salt, the compound
or salt will be substantially entrained within the hydrophilic
center or core of the liposomes. The lipid layer employed may be of
any conventional composition and may either contain cholesterol or
may be cholesterol-free. When the compound or salt of interest is
water-insoluble, again employing conventional liposome formation
technology, the salt may be substantially entrained within the
hydrophobic lipid bilayer that forms the structure of the liposome.
In either instance, the liposomes that are produced may be reduced
in size, as through the use of standard sonication and
homogenization techniques. The liposomal formulations containing
the progesterone analogue or salts thereof, may be lyophilized to
produce a lyophilizate which may be reconstituted with a
pharmaceutically acceptable carrier, such as water, to regenerate a
liposomal suspension.
[0529] Pharmaceutical formulations are also provided which are
suitable for administration as an aerosol, by inhalation. These
formulations comprise a solution or suspension of the desired
steroid analogue or a salt thereof or a plurality of solid
particles of the compound or salt. The desired formulation may be
placed in a small chamber and nebulized. Nebulization may be
accomplished by compressed air or by ultrasonic energy to form a
plurality of liquid droplets or solid particles comprising the
compounds or salts.
[0530] In another embodiment, the compounds are prepared with
carriers that will protect them against rapid elimination from the
body, such as a controlled release formulation, including implants
and microencapsulated delivery systems. Biodegradable,
biocompatible polymers can be used, such as ethylene vinyl acetate,
polyanhydrides, polyglycolic acid, collagen, polyorthoesters and
polylactic acid. Methods for preparation of such formulations will
be apparent to those skilled in the art.
VII. Synthesis of the Active Compounds
[0531] General schemes 1-8 below describe the preparation of
selected steroid analogues. It is understood that the specific
synthetic steps are not limited to the reactions shown in the
schemes and that many alternative reaction sequences known in the
art are suitable for the preparation of the analogues. Furthermore,
it is understood that any naturally occurring or synthetic amino
acid in the D, L or D,L configuration may be used. The invention is
not limited by the type of protecting group and any suitable
protecting group may be used. Protecting groups for amino groups
and ketone groups are well known in the art and described by Greene
et al. "Protective Groups in Organic Synthesis," John Wiley and
Sons, 4.sup.th edition, 2007.
Progesterone Analogues Substituted at the 3-Position:
[0532] Compounds derivatized at the 3-position of the steroid ring
system to comprise an ester of an amino acid may be prepared using
the general process described in Scheme 1 below. Starting from
progesterone, the carbonyl group at the 3-position is selectively
reduced to produce the allylic alcohol 2. One example of a
selective reduction is the Luche Reduction which use sodium
borohydride and cerium trichloride in methanol (see Luche, J.-L. J.
Am. Chem. Soc., 1978, 100, 2226). Typically, the reduction is run
at low temperature to avoid over-reduction of the C-20 carbonyl.
Alcohol 2 is then esterified with a suitable amino acid derivative
form the progesterone analogue 3. The protecting group is removed
and a suitable salt, such as an HCl salt, may be formed, if
desired.
##STR00088##
[0533] In one embodiment, the ester bond may be formed by reaction
of the hydroxyl group of 2 with a protected amino acid acyl halide,
where X is chloro, bromo, iodo or fluoro. In another embodiment,
the ester bond may be formed by reacting the hydroxyl group with an
activated carboxylic acid, where X is an activated leaving group.
Many reagents are known that will activate carboxyl groups to react
with nucleophiles. For example, a variety of peptide coupling
reagents well known in the art are used to activate carboxyl groups
in-situ to react with amino groups of amino acids to form peptide
bonds. These reagents can also activate carboxylic acids to form
reactive intermediates that will react with hydroxy groups on the
steroid compound. Non-limiting examples of the carboxyl activating
groups include carbodiimide reagents, phosphonium reagents such as
benzotriazolyloxy-tris-(dimethylamino) phosphonium
hexafluorophosphate (BOP) and the like, uronium or carbonium
reagents such as O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (HBTU),
benzotriazol-1-yl-oxy-tripyrrolidinophosphonium hexafluorophosphate
(PyBOP) and the like; 1-ethoxycarbonyl-2-ethoxy-1,2-dihydroqunoline
(EEDQ); 1-methyl-2-chloropyridinium iodide (Muikaiyama's reagent)
and the like. In other embodiments, the ester may be formed by
trans-esterification of another ester group including active esters
such as ap-nitrophenyl ester, a pentafluorophenylester, an
N-hydroxysuccinimidyl ester, a 1-hydroxybenzotriazolyl ester and
the like. An acyl azide group may also be used to form the ester
bond. In another embodiment, the ester may also be formed by
reaction of the hydroxy with a symmetric or mixed anhydride (X is
RC(O)O--). Catalysts such as 4-dimethylaminopyridine (DMAP) and the
like may be used to facilitate the ester formation.
Progesterone Analogues Substituted at the 20-Position:
[0534] Scheme 2 below illustrates the general synthetic process for
the formation of steroid analogues that comprise amino acid
residues at the 20-position of the ring. In this process,
progesterone is reduced to the diol using a strong reducing
reagent, such as lithium aluminum hydride. The allylic hydroxyl
group is then selectively oxidized to produce the enone 4, with the
C-20 hydroxyl group intact. Any suitable oxidizing agent that will
selectively oxidize an allylic alcohol may be used. One
non-limiting example is manganese dioxide (MnO.sub.2). The
resulting alcohol 3 is then esterified to produce the desired
steroid analogue 5 comprising an amino acid residue at the
20-position. As described above for Scheme 1, the esterification
reaction may be accomplished with a variety of reagents, including
a protected amino acid halide or with a protected amino acid using
a coupling reagent known in the art to activate carboxylate
groups.
##STR00089##
C-3 Progesterone --C.dbd.NR Derivatives
[0535] Scheme 3 below illustrates the preparation of an analogue of
Formula VII, substituted at C-3 with the group .dbd.N--R.sup.3
where R.sup.3 is --OR.sup.11, and R.sup.11 is an amino acid
residue. The C20 carbonyl is first protected with a suitable
protecting group such as the cyclic ketal 6 to prevent reaction
with the nucleophilic hydroxylamine. The remaining enone is reacted
with hydroxylamine to produce a mixture of E/Z 7a and 7b. The
E-oxime 7a is then esterified with a suitably protected amino acid
halide or using a protected amino acid with a coupling reagent as
described above for Scheme 1 to produce the E-isomer of protected
analogue 8a. Removal of the cyclic acetal under acidic conditions
followed by removal of the amino protecting group under typically
basic conditions provides the C3 analogue 9a, which is converted to
the hydrochloride salt upon treatment with HCl. The corresponding
Z-isomers 8b and 9b are prepared using Z-oxime 7b in the same
manner.
##STR00090##
Scheme 4 shows the general synthesis of an analogue of Formula IV
substituted at C-3 with the group R.sup.5--N.dbd.C where R.sup.5 is
R--NH--, and R is an amino acid residue. Starting from protected
intermediate 6, the C-3 carbonyl is reacted with hydrazine to
produce the hydrazone 10. The hydrazone is then reacted with a
suitable reactive amino acid derivative as described above for
Scheme 1 to yield the hydrazide 11. The hydrazide may be converted
to a pharmaceutically acceptable salt by treatment with a
pharmaceutically acceptable acid, such as HCl.
##STR00091##
C-3 Pregnanolone and Allopregnanolone Derivatives
[0536] Scheme 5 below shows the preparation of allopregnanolone
(ALLO) analogues substituted at C-3 with an amino acid residue.
Pregnenolone is first reduced with hydrogen catalyzed by palladium
on carbon to produce compound 12 in the 3-beta, 5-alpha
configuration. Compound 12 is then esterified as described for
Scheme 1 above with a reactive protected amino acid reagent
followed by deprotection to produce compound 13 substituted at the
C-3 position with an amino acid residue. The HCl salt is formed by
treatment with HCl as before.
[0537] To produce compound 15, in which the amino acid substituent
has the opposite stereoisomeric configuration at C-3, the stereo
configuration of the hydroxyl group in compound 12 is inverted
using Mitsunobu conditions (see Mitsunobu et al., Bull. Chem. Soc.
Japan 1967, 40, 2380-2382 and Mitsunobu et al., Synthesis 1981,
1-28 and Castro et al., Org. React. 1983, 29, 1) to form compound
14 with the 3-alpha, 5-alpha configuration. Compound 14 is
esterified as described above to produce compound 15 with the
3-alpha, 5-alpha configuration, followed by treatment with HCl to
form the salt.
##STR00092##
[0538] Scheme 6 below shows a general process for the preparation
of C-3 substituted pregnanolone analogues in different
stereoisomeric configurations. Starting from progesterone,
reduction of the enone with hydrogen under palladium on carbon
forms compound 16. Reduction of the cyclic ketone, using a suitable
reducing agent such as sodium borohydride, forms a mixture of
alcohols 17a and 17b. Esterification of alcohols 17a and 17b
followed by removal of the protecting group and salt formation
provides pregnanolone analogues 18a and 18b.
##STR00093##
[0539] Steroid analogues with a double bond between the C5 and C6
positions may be prepared according to the general process shown in
Scheme 7 below.
##STR00094##
[0540] Esterification of pregnenolone with a suitably protected
amino acid as described for scheme 1 above provides compound 19,
with an amino acid residue at the 3-position. Protection of the
hydroxyl of pregnenolone followed by reaction with hydroxylamine
provides the E- and Z-isomers 20a and 20b. If desired, the isomers
may be separated at this stage. Reaction of 20a and 20b with a
suitably protected amino acid, followed by deprotection and
treatment with HCl provides compounds 21a and 21b.
[0541] Modification of the process shown in Scheme 7 leads to
related compounds with a double bond between C5 and C6. For
example, to obtain the steroid analogues corresponding to compounds
21a and 21b in which the C-3 hydroxyl is in the ketone oxidation
state, compounds 20a and 20b may be deprotected to the alcohol and
oxidized to form the ketone prior to reaction with the activated
amino acids reagent. Reduction of compound 19 will provide the
corresponding C20 alcohol, which may be esterified as described
above to form an analogue substituted at both C3 and C20
positions.
[0542] Reduction of protected pregnenolone followed by
esterification of the resulting C20 hydroxyl group with a suitably
protected activated amino acid will provide the C20 amino acid
substituted derivative after removal of the protecting group.
[0543] Protection of the C20 ketone, for example as a cyclic ketal,
followed by oxidation of the C3 hydroxyl to the corresponding
ketone and then reaction with hydroxyl amine will provide the
corresponding C3 oximes, which can be reacted with suitably
protected activated amino acids to prepare the
.dbd.N--NR.sup.11R.sup.12 compounds.
[0544] Analogues with a double bond between the C1 and C2 carbons
may be prepared according to the process depicted in Scheme 8
below.
##STR00095##
[0545] Starting from protected compound 6, treatment with a bulky
base such as lithium diisopropylamide (LDA) or the like, to form
the enolate species, followed by reaction with a suitable source of
electrophilic selenium, such as diphenyldiselenide, provides
compound 22. Treatment of compound 22 with a suitable oxidizing
agent, such as hydrogen peroxide, provides compound 24, which is
deprotected to provide compound 25.
Enantiomeric Progesterone Compounds
[0546] In one embodiment, the invention provides enantiomeric
progesterone and neuroprotective steroid compounds. The enantiomer
of progesterone (ent-PROG) has shown similar efficacy to
progesterone and allopregnanolone across several measures relevant
to neuroprotection, including the reduction of cerebral edema,
reduction of pro-inflammatory cytokine expression, and reduction in
proapoptotic p53 protein expression. Ent-PROG treatment was also
shown to result in significantly increased glutathione reductase
activity, a measure of its potential to minimize oxidative stress
following TBI, relative to both progesterone and allopregnanolone.
Although it binds with moderate affinity to the classical
progesterone receptor (PR), ent-PROG does not activate PR-mediated
gene transcription. Thus it is thought that ent-PROG is able to
achieve its neuroprotective effects either through
transcription-independent PR-mediated signaling or via
PR-independent pathways. In light of these findings, and with the
goal of providing a compound of improved efficacy relative to PROG
or allopregnanolone, the development of a complementary set of
ent-PROG based analogue compounds was pursued.
[0547] Addition of methyl vinyl ketone (MVK) to
2-methyl-1,3-cyclopentadione (37, Scheme 9) gave the triketone 38.
The organocatalyst D-proline was then used in order to achieve
asymmetric cyclization of 38 to give the Hajos-Parrish ketone (39).
Sodium borohydride reduction of 39 was followed by protection of
the newly formed secondary alcohol 40 as its tert-butyl ether (41).
Introduction of an .alpha.-methylene group was achieved through
initial carbonation of 41 with Stiles' reagent, methyl magnesium
carbonate (MMC), in DMF. Selective reduction of the C-4-C-5 double
bond of compound 43 was immediately followed by decarboxylation of
the unstable saturated intermediate 44 to give the enone 45 with
trans ring junction.
##STR00096##
[0548] Synthesis of the .beta.-keto ester annulating agent 50 began
with ketalization of ethyl-5-oxohexanoate and subsequent reduction
of the ester 47 with LiAlH4 to give alcohol 48 (Scheme 10). Swern
oxidation of 48 was followed by tin(II) chloride catalyzed coupling
with ethyl diazoacetate to give the P-keto ester 50.
##STR00097##
[0549] Michael addition of 6-keto ester 50 to enone 45, along with
in situ Robinson annulation, saponification, and finally
decarboxylation, gave the BCD ring system 51. Reductive alkylation
served to introduce what would become the C-19 methyl group of
ent-PROG. Reflux of 52 overnight in methanolic HC1 gave
ent-testosterone (53). Ent-testosterone was then prepared as the
C-3 ketal 54 and the C-17 secondary alcohol was oxidized using
pyridinium chlorochromate (PCC) to give ketone 55. Treatment of 55
with ethyltriphenylphosphonium bromide under Wittig conditions gave
the alkene 56. A final three step sequence involving hydroboration,
oxidation, and acid catalyzed removal of the ketal was carried out
without intermediate purification steps to give ent-PROG (57) in
good yield.
##STR00098##
[0550] Luche reduction of ent-PROG gave the C-3 a-hydroxy compound
58 (Scheme 12). The same series of reactions involving amino acid
coupling, Fmoc cleavage, and HCl salt formation that had been
developed for the C-3 nut-PROG series of compounds was applied here
to give the ent-PROG derivative P2-13. Additional neuroprotective
analogues derived from ent-PROG are prepared according to the
description provided above and in the following examples.
##STR00099##
[0551] The present invention will be understood more readily by
reference to the following examples, which are provided by way of
illustration and are not intended to be limiting of the
invention.
EXAMPLES
Example 1
Effectiveness of Progesterone Analogues in Reducing Post-Injury
Edema
[0552] A well established whole animal model of TBI was employed in
order to investigate the potential efficacy of the PROG analogue
compounds relative to PROG in reducing cerebral edema following
injury. Anesthetized male rats were first subjected to cortical
contusion and were then given two 8 mg/kg doses of the test
compound, the first at 1 h post-surgery and the second at 6 h. The
animals were then sacrificed and tissue samples were taken from
both injured and non-injured sections of the brain. Wet and dry
weights were collected for each sample and cerebral edema (% water
content) was determined as the difference in wet and dry weights
divided by the wet weight. A "% mean difference" value could then
be calculated based on the relative edema difference between
injured and non-injured tissue samples for a given animal. The
"sham" animals did not receive an injury but served as a control
group for possible anesthesia and stress factors. The "vehicle"
group were subjected to cortical injury but received only the drug
carrier (22.5% 2-hydroxypropyl-.beta.-cyclodextrine in water). All
experimental treatments given by injection were made in stock
solutions using 2-Hydroxypropyl-b-cyclodextrin (HBC; 45% w/v
solution in H.sub.2O) as the solvent. These experimental solutions
were then diluted 1:1 with sterile water for a final concentration
of HBC of 22.5%. Several of the analogues showed equivalent
efficacy to progesterone in the cerebral edema assay, including the
valine tethered C-3-.beta.-hydroxy PROG derivative P1-31 and the
oxime based prodrug compound P1-185. Compound P1-131, the valine
coupled derivative of ALLO itself, showed the greatest edema
reduction among the ALLO isomer group. Perhaps most notably though
was the activity of oxime prodrug P1-186, which showed an average
reduction in edema levels almost twice that of PROG.
Results:
[0553] FIG. 1 shows the % difference edema results for brain tissue
24 hours post brain injury. The mean % difference calculated for
sham, vehicle, progesterone, Compound 31, Compound 57 and Compound
79 subjects were 0.6%, 1.2%, 2.0%, 2.2%, 3.3% and 1.9%,
respectively. Samples treated with progesterone, and Compounds 31,
57 and 79 all showed a decrease in edema compared to subjects
treated with vehicle.
Example 2
Preparation of Compounds General Experimental
[0554] All reagents were obtained from Aldrich. Reactions requiring
anhydrous conditions were performed in oven-dried glassware under
dry argon. All solvents used were anhydrous or kept dry over
activated 4 .ANG. molecular sieves. Convection was achieved by use
of a magnetic stirring bar unless otherwise noted. The following
abbreviations may be used: dichloromethane (DCM), diethyl ether
(ether), water (DI), hexane (hex), ethyl acetate (ea),
dimethylformamide (DMF), acetonitrile (ACN), tetrahydrofuran (THF),
round bottomed flask (RBF), hours (h), minutes (min), millimole
(mmol), equivalents (eq). Reaction progress was monitored via
thin-layer chromatography (TLC) on pre-coated glass-backed plates
(silica gel 60 .ANG. F.sub.254, 0.25 mm thickness) purchased from
EM Science. Flash chromatography was carried out with silica gel 60
.ANG. (230-400 mesh) from Sorbent Technologies. Automated
chromatography was performed on an Isco Combiflash Companion.
Unless otherwise stated, organic extracts were dried over
commercially available magnesium sulfate and the solvents were
removed by rotary evaporation. Brine refers to a saturated sodium
chloride solution. .sup.1H and .sup.13C NMR spectra were recorded
on either a 400 MHz Inova spectrometer or 600 MHz Inova
spectrometer in deuterated chloroform (CDCl.sub.3) and referenced
to the residual solvent peak (.sup.1H .delta. 7.27 ppm, .sup.13C
.delta. 77.23 ppm). Chemical shifts are reported in parts per
million (.delta.), and coupling constants are reported in hertz
(Hz). The following abbreviations will be used: singlet (s),
doublet (d), triplet (t), quartet (q), multiplet (m). Mass spectra
were obtained on either a VG 70-S Nier Johnson or JEOL Mass
Spectrometer. Elemental analyses were performed by Atlantic
Microlab (Norcross, Ga.).
[0555] The abbreviations PROG and ALLO are used for progesterone
and allopregnanolone, respectively. Scheme 13 below depicts the
synthesis of various C3 amino acid derivatives of the invention,
along with representative reaction conditions and reagents.
TABLE-US-00003 Scheme 13 ##STR00100##
Example 3
C-3 Progesterone Derivatives
##STR00101##
[0556] 3-.beta.-Hydroxy-progesterone (2)
[0557] Progesterone (3.14 g, 10.0 mmol) was added with cerium
chloride heptahydrate (3.73 g, 10.0 mmol, 1.00 eq) to an oven dried
three necked 250 mL RBF with thermometer. Methanol (100 mL) was
added under argon and the solution was chilled to -20.degree. C.
Sodium borohydride (0.189 g, 5.00 mmol, 0.500 eq) was then added in
bulk. Solution temperature raised briefly up to -16.degree. C.
After 15 minutes, 37 mL acetone was added and the solution was
warmed to ambient temperature. Water (25 mL) was added and the
solvent volume was reduced by approximately 100 mL. Ether was
added, along with more water, which caused the solution to become
clear and colorless. The aqueous layer was extracted with ether.
The organic layers were combined, washed with brine, dried,
filtered, and concentrated to give 3.14 g white solid. The solid
was prepared as a silica cake, loaded onto a 500 mL silica column,
and eluted with 3 L 20% ethyl acetate in hexanes, followed by 2 L
25% ethyl acetate in hexanes. Initially eluting pure fractions were
combined and concentrated to give 1.56 g white solid that was 90%
pure as determined by proton NMR (other 10% was progesterone).
(44%) white solid; R.sub.f=0.38 (1:1 EA/hex, PMA stain); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 5.29 (d, 1H, J=1.6 Hz), 4.18-4.12
(m, 1H), 2.51 (t, 1H, J=8.8 Hz), 2.25-0.77 (m, 20H), 2.11 (s, 3H),
1.04 (s, 3H), 0.62 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 209.9, 147.4, 123.8, 68.1, 63.9, 56.5, 54.5, 44.3, 39.1,
37.5, 36.1, 35.6, 33.1, 32.3, 31.7, 29.6, 24.6, 22.9, 21.2, 19.1,
13.6.
##STR00102##
N-Fmoc-L-valine-3-.beta.-progesterone (Fmoc 3a)
[0558] An oven dried 50 mL RBF was charged with 90%
3-beta-hydroxy-progesterone (0.352 g, 1.00 mmol), N-Fmoc-L-valine
(0.339 g, 1.00 mmol, 1.00 eq), and dimethylaminopyridine (DMAP)
(0.0244 g, 0.200 mmol, 0.200 eq). The flask was sealed, evacuated,
and inert gas flushed and 15 mL anhydrous dichloromethane was
added, followed by addition of 1.10 mL (1.10 mmol, 1.10 eq) 1 M
dicyclohexylcarbodiimide (DCC) in dichloromethane. The solution was
stirred overnight then filtered through Celite. The filtrate was
concentrated, prepared as a silica cake and eluted on a 40 g silica
column with a 0-25% ethyl acetate in hexanes gradient. The main
product was isolated as 0.554 g (87%) clear oil that foamed on
drying. R.sub.f=0.40 (1:1 EA/hex, PMA stain); .sup.1H NMR (600 MHz,
CDCl.sub.3) .delta. 7.78 (d, 2H, J=7.2 Hz), 7.63-7.61 (m, 2H), 7.41
(t, 2H, J=7.2 Hz), 7.33 (t, 2H, J=7.2 Hz), 5.35 (d, 1H, J=9.0 Hz),
5.31 (t, 1H, J=7.8 Hz), 5.21 (s, 1H), 4.40 (d, 2H, J=7.2 Hz), 4.31
(dd, 1H, J=9.0, 4.2 Hz), 4.25 (t, 1H, J=7.2 Hz), 2.52 (t, 1H, J=9.0
Hz), 2.23-2.16 (m, 3H), 2.12 (s, 3H), 2.05-1.96 (m, 3H), 1.78-1.55
(m, 6H), 1.50-1.33 (m, 4H), 1.25-1.10 (m, 2H), 1.06 (s, 3H), 1.00
(d, 3H, J=7.2 Hz), 0.93 (d, 3H, J=7.2 Hz), 0.90-0.79 (m, 2H), 0.64
(s, 3H).
##STR00103##
3-.beta.-L-Valine-progesterone (3a)
[0559] A 25 mL RBF was charged with 0.340 g (0.533 mmol) compound
3a. The flask was evacuated and inert gas flushed and 5 mL each of
acetonitrile and dimethylformamide were added. A 0.527 mL (5.33
mmol, 10.0 eq) volume of piperidine was added and the clear
colorless solution was stirred at room temperature for 30 min. The
solvent was removed with addition of toluene for complete removal
of DMF. A white solid formed that was redissolved in a minimum
amount of toluene and loaded neat onto a 12 g silica column and
eluted with 0-75% ea in hexanes. Main product containing fractions
were combined and dried to give 0.196 g (89%) white foam. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 5.29-5.23 (m, 1H), 5.20 (d, 1H,
J=1.6 Hz), 3.27 (d, 1H, J=4.8 Hz), 2.52 (t, 1H, J=9.2 Hz),
2.36-1.93 (m, 6H), 2.11 (s, 3H), 1.79-1.08 (m, 14H), 1.06 (s, 3H),
0.98 (d, 3H, J=6.8 Hz), 0.95-0.77 (m, 3H), 0.90 (d, 3H, J=6.8 Hz),
0.62 (s, 2H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.8,
175.6, 149.3, 119.3, 71.3, 63.8, 60.2, 56.4, 54.2, 44.3, 39.0,
37.5, 36.0, 35.2, 33.0, 32.3 (2C), 31.7, 25.3, 24.6, 22.9, 21.1,
19.6, 19.0, 17.3, 13.6; IR (solid): 2934, 2843, 1724, 1705, 1384,
1354, 1166, 1146, 978, 873, 852 cm.sup.-1; HRMS-ESI m/z 416.3156
([M+H].sup.+, C.sub.26H.sub.42NO.sub.3 requires 416.3159).
##STR00104##
3.beta.-L-Valine-progesterone-HCl (P1-31)
[0560] A 10 mL RBF with stir bar was charged with 83 mg compound 4
and the flask was evacuated and flushed with argon. Anhydrous ether
(2 mL) was added and the solution was chilled in an ice bath.
Hydrogen chloride solution (0.10 mL 2.0 M in ether, 0.20 mmol, 1.0
eq) was added dropwise. A white precipitate formed in solution. The
precipitate was filtered and washed with chilled ether. The product
was recovered as 68 mg (75%) off-white solid.
[0561] The following compounds were prepared by the methods as
described above:
##STR00105##
3.beta.-L-Alanine-progesterone-HCl (P1-29)
[0562] (52% from P1-30) white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.63 (bs, 2H), 5.25 (t, 1H, J=7.6 Hz), 5.14 (s,
1H), 4.12 (d, 1H, J=7.6 Hz), 2.64 (bs, 1H), 2.43 (t, 1H, J=8.8 Hz),
2.20-0.68 (m, 21H), 2.04 (s, 3H), 1.65 (d, 3H, J=7.2 Hz), 0.99 (s,
3H), 0.57 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta.
209.7, 170.1, 150.1, 118.4, 73.4, 63.8, 56.4, 54.1, 49.5, 44.2,
39.0, 37.5, 36.0, 35.0, 33.0, 32.3, 31.7, 25.0, 24.6, 23.0, 21.2,
19.0, 16.4, 13.5; hR (film): 2934, 2849, 1741, 1703, 1237, 1207,
1113, 916, 731 cm'; HRMS-ESI m/z 388.2847 ([M-Cl].sup.f,
C.sub.24H.sub.38NO.sub.3 requires 388.2846); Anal. Calcd for
C.sub.24H.sub.38ClNO.sub.3+%2H.sub.20: C, 66.57; H, 9.08; N, 3.23.
Found C, 66.42; H, 9.01; N, 3.19.
##STR00106##
3.beta.-L-Phenylalanine-progesterone-HCl (P1-32)
[0563] (51% from P1-30) white solid. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 8.71 (bs, 2H), 7.30-7.21 (m, 5H), 5.17 (bs,
1H), 4.97 (s, 1H), 4.32 (bs, 1H), 3.47-3.29 (m, 2H), 2.48 (t, 1H,
J=8.6 Hz), 2.20-0.68 (m, 20H), 2.10 (s, 3H), 0.97 (s, 3H), 0.60 (s,
3H); .sup.13CNMR (100 MHz, CDCl.sub.3) .delta. 209.7, 168.8, 149.9,
134.2, 130.1, 129.0, 127.8, 118.2, 73.6, 63.8, 56.4, 54.5, 54.1,
44.2, 39.0, 37.4, 36.6, 36.0, 34.8, 33.0, 32.2, 31.7, 24.9, 24.6,
23.0, 21.2, 18.9, 13.6; IR (film): 2929, 2848, 1732, 1701, 1233,
1202, 1109, 912, 729, 700 cm.sup.-1; HRMS-ESI m/z 464.3160
([M-Cl]+, C.sub.30H.sub.42NO.sub.3 requires 464.3159); Anal. Calcd
for C.sub.30H.sub.42ClNO.sub.3+1/2H.sub.20: C, 70.77; H, 8.51; N,
2.75. Found C, 70.74; H, 8.31; N, 2.78.
##STR00107##
3.beta.1-L-Proline-progesterone-HCl (P1-33)
[0564] (38% from P1-30) white solid. .sup.1H NMR (400 MHz, DMSO)
.delta. 10.32 (bs, 1H), 9.02 (bs, 1H), 5.26 (s, 1H), 5.22 (s, 1H),
4.33 (s, 1H), 3.40 (s, 1H), 3.20 (d, 2H, J=7.2 Hz), 2.56 (t, 1H,
J=8.6 Hz), 2.30-0.73 (m, 23H), 2.05 (s, 3H), 1.02 (s, 3H), 0.54 (s,
3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.6, 168.9,
150.6, 118.0, 74.1, 63.8, 59.5, 56.4, 54.0, 46.6, 44.2, 38.9, 37.5,
35.9, 34.8, 33.0, 32.6, 31.7, 29.4, 25.1, 24.5, 24.0, 22.9, 21.1,
19.0, 13.5; IR (film): cm.sup.-1; HRMS-ESI m/z 414.3005
([M-Cl].sup.+, C.sub.26H.sub.40NO.sub.3 requires 414.3003); Anal.
Calcd. for C.sub.26H.sub.40ClNO.sub.3+1/2H.sub.20: C, 68.03; H,
9.00; N, 3.05. Found C, 68.21; H, 8.89; N, 3.02.
##STR00108##
3.beta.-L-Tryptophan-progesterone-HCl (P1-34)
[0565] (43% from P1-30) white solid. .sup.1H NMR (400 MHz, DMSO)
.delta. 11.1 (bs, 1H), 8.62 (bs, 2H), 7.53 (d, 1H, J 7.6 Hz), 7.36
(d, 1H, J=7.6 Hz), 7.24 (s, 1H), 7.08 (t, 1H, J=7.4 Hz), 6.99 (t,
1H, J 7.4 Hz), 5.07 (bs, 1H), 4.73 (s, 1H), 4.14 (bs, 1H),
3.46-3.20 (m, 2H), 2.54 (t, 1H, J=8.8 Hz), 2.20-0.67 (m, 20H), 2.05
(s, 3H), 0.95 (s, 3H), 0.53 (s, 3H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 209.7, 169.3, 150.3, 136.4, 126.9, 126.6,
122.0, 119.3, 118.5, 118.1, 112.1, 105.9, 74.1, 63.8, 56.4, 54.0,
53.6, 44.2, 39.0, 37.5, 35.9, 34.7, 33.0, 32.3, 31.7, 26.1, 25.0,
24.6, 23.0, 21.1, 19.0, 13.6; IR (film): 2929, 2849, 1.732, 1701,
1456, 1435, 1354, 1218, 1108, 730 cm.sup.-1; HRMS-ESI m/z 503.3271
([M-Cl].sup.+, C.sub.32H.sub.43N.sub.20.sub.3 requires 503.3268);
Anal. Calcd for C.sub.32H.sub.43ClN.sub.20.sub.3+.sup.3/4H.sub.20:
C, 69.54; H, 8.12; N, 5.07. Found C, 69.57; H, 8.06; N, 5.04.
[0566] Scheme 14 below depicts the synthesis of C3 valine
derivative, where the C3 hydroxy group is in the .beta.
configuration. Similar conditions may be used for the synthesis of
other amino acid derivatives.
##STR00109##
##STR00110##
3.alpha.-Hydroxy-progesterone (26)
[0567] Ester formation: Compound 12 (0.560 g, 1.59 mmol),
triphenylphosphine (0.877 g, 3.34 mmol, 2.10 eq), and
p-nitrophenylbenzoic acid (0.559 g, 3.34 mmol, 2.10 eq) were added
to a 100 mL oven dried flask that was evacuated and inert gas
flushed. A 20 mL volume of anhydrous THF was added. The solution
was cooled to 0.degree. C. and diisopropylazodicarbonate (0.693 mL
in 10 mL THF solution, 3.34 mmol, 2.10 eq) was added dropwise over
1 h. The solution was stirred at 0.degree. C. for an additional 1
h. The reaction was diluted with 50 mL ether and washed with
saturated sodium bicarbonate solution (3.times.25 mL). The aqueous
layers were combined and extracted with ether. The organic layers
were combined, dried, filtered, and concentrated to give a pale
orange oil. The oil was loaded onto a 40 g silica column in a
minimum amount of DCM and eluted with 0-25% ea in hex over 35 min.
Main peak fractions were combined and concentrated to give a clear
oil that crystallized on standing. Crude wet mass was 0.820 g.
Saponification: The esterification product (0.613 g, 1.32 mmol) was
dissolved in 12 mL 5:3 MeOH/THF in a 25 mL RBF and potassium
carbonate (0.364 g, 2.63 mmol, 2.00 eq), dissolved in 2 mL DI, was
added. A white precipitate formed in solution that gradually
dissolved over the course of 1 h. The solvents were removed and the
aqueous layer was extracted with ether. The organic layers were
combined, washed with brine, dried, filtered, and concentrated. The
resulting white solid was re-dissolved in a minimum amount of DCM
and eluted on a 40 g silica column with 0-35% ea in hexanes
gradient over 40 min. The desired major product was isolated as
0.380 g (91%) white solid. R.sub.f=0.31 (1:1 EA/hex, PMA stain);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.47 (dd, 1H, J=4.8, 1.6
Hz), 4.10-4.06 (m, 1H), 2.53 (t, 1H, J=9.2 Hz), 2.27-0.82 (m, 20H),
2.12 (s, 3H), 0.99 (s, 3H), 0.64 (s, 3H); .sup.13C NMR. (100 M.Hz,
CDCl.sub.3) .delta. 209.8, 150.1, 121.1, 64.4, 63.9, 56.5, 54.1,
44.4, 39.2, 37.7, 36.0, 32.9, 32.5, 31.9, 31.7, 28.0, 24.6, 23.0,
21.7, 18.3, 13.6; TR (solid): 3485, 3414, 2929, 2842, 1694, 1356,
1.015 cmi.sup.-1 HRMS-ESI m/z 299.2366 ([M+H-H.sub.2O].sup.+,
C.sub.21H.sub.31O requires 299.2369).
##STR00111##
N-Fmoc-L-valine-3a-progesterone (Fmoc P1-163)
[0568] Compound 14 (0.100 g, 0.314 mmol), N-Fmoc-valine (0.213 g,
0.628 mmol, 2.00 eq),
N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCI)
(0.120 g, 0.628 mmol, 2.00 eq), and DMAP (4 mg, 0.10 eq) were added
to an oven dried 25 mL RBF. The flask was evacuated and inert gas
flushed and 6 mL anhydrous DCM was added. The reaction was stirred
at room temperature for 36 h. The solution was quenched and washed
with saturated ammonium chloride solution (2.times.20 mL). The
aqueous layers were combined and extracted with DCM. The organic
layers were combined, washed with brine, dried, filtered, and
concentrated. The clear oil was loaded in a minimum amount of DCM.
onto a 12 g silica column and eluted with a 0-15% ea in hex
gradient over 45 minutes. The desired product was obtained as 0.193
g (96%) white foam. R.sub.f=0.63 (1:1 EA/hex); .sup.1H NVIR (400
MHz, CDCl.sub.3) 6 7.77 (d, 211, J 7.6 Hz), 7.63 (d, 2H, J 7.2 Hz),
7.41 (t, 2H, J=7.2 Hz), 7.33 (dt, 2H, J=7.2, 0.8 Hz), 5.44 (d, 1H,
J=4.4 Hz), 5.36 (d, 1H, J=8.8 Hz), 5.19 (d, 1H, J=2.4 Hz),
4.48-4.44 (m, 1H), 4.35-4.23 (m, 3H), 2.43 (t, 1H, J=8.8 Hz),
2.28-0.79 (m, 20H), 2.09 (s, 3H), 1.00 (d, 3H, J=6.8 Hz), 0.99 (s,
3H), 0.94 (d, 3H, J=7.2 Hz), 0.62 (s, 3H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 209.8, 172.0, 156.5, 152.9, 144.2, 143.9,
141.5, 127.9, 127.3, 125.5, 125.4, 120.2, 116.5, 69.0, 67.2, 63.8,
59.2, 56.2, 53.9, 47.5, 44.2, 38.9, 37.6, 35.8, 32.5, 31.7, 31.6,
25.1, 24.5, 22.9, 21.6, 19.3, 1.8.1, 17.6, 13.5; IR (solid): 3335,
2935, 1700, 1449, 1237, 1195, 759, 740 cm.sup.-1
##STR00112##
3.alpha.-L-Valine-progesterone (PI163 Free Base)
[0569] Prepared according to the method described for compound 5b.
(85%) clear oil; .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 5.44 (d,
1H, 1-4.8 Hz), 5.17 (s, 1H), 3.28 (d, 111, J=4.8 Hz), 2.53 (t, 1H,
J=9.0 Hz), 2.27-0.86 (m, 2211), 2.12 (s, 311), 1.01 (s, 3H), 1.00
(d, 3H, J=6.6 Hz), 0.91 (d, 3H, J=7.2 Hz), 0.65 (s, 3H); .sup.13C
NMR (150 MHz, CDCl.sub.3) .delta. 209.8, 175.5, 152.5, 117.1, 68.1,
63.9, 60.1, 56.4, 54.0, 44.3, 39.1, 37.6, 35.9, 32.6, 32.5 (2C),
32.4, 31.7, 25.2, 24.6, 23.0, 21.6, 19.6, 18.2, 17.3, 13.6; IR
(solid): 2933, 2872, 1722, 1704, 1383, 1358, 1237, 1178, 976
cm.sup.-1; H12MS-ESI m/z 416.3155 ([.M+H].sup.+,
C.sub.26H.sub.42NO.sub.3 requires 416.3159).
##STR00113##
3.alpha.-L-Valine-progesterone-HCl (P1-163)
[0570] Prepared according to the method described for compound
P1-31. (21%) white powdery solid; .sup.1H NMR (400 MHz, CDCl.sub.3)
5.45 (d, 1H, J=4.4 Hz), 5.25 (s, 111), 3.82 (d, 1H, J=3.2 Hz), 2.53
(t, 1TI, J-=9.2 Hz), 2.48-2.37 (m, 1H), 2.29-0.79 (m, 22H), 2.12
(s, 3H), 1.15 (d, 3H, J=6.8 Hz), 1.11. (d, 3H, J=6.8 Hz), 1.00 (s,
3H), 0.64 (s, 3H); .sup.13C NMR. (100 MHz, CDCl.sub.3) .delta.
209.8, 169.7, 153.3, 116.3, 70.0, 63.9, 59.0, 56.3, 53.9, 44.3,
39.0, 37.6, 35.9, 32.5, 32.4 (2C), 31.7, 30.6, 25.0, 24.6, 23.0,
21.6, 18.7, 18.2 (2C), 13.6; IR (film): 2935, 2876, 2620, 1734,
1703, 1383, 1357, 1228, 731 cm.sup.-1; Anal. Calcd for
C.sub.26H.sub.42CINO.sub.3+1/2H.sub.2O: C, 67.73; H, 9.40; N, 3.04.
Found C, 67.00; H, 9.53; N, 3.04.
Example 4
C-20 Progesterone Derivatives
[0571] Scheme 15 describes the synthesis of C20 valine derivative
P1-57 with representative reagents and conditions. This procedure
may be used to prepare C20 derivatives of other amino acid using
the same or alternate reagents.
##STR00114##
3,20-Hydroxy-progesterone (4a)
[0572] An oven dried RBF was charged with 25 mL anhydrous THF and
chilled in an ice bath. A 4.50 mL volume (9.00 mmol, 2.25 eq) of
2.0 M lithium aluminum hydride in THF was added. A separate
.about.10 mL solution of progesterone (1.26 g, 4.00 mmol) in
anhydrous THF was prepared in a dry flask. The solution was
transferred to the reaction flask dropwise over 30 minutes. The
mixture was heated under reflux for 1 h, cooled to room
temperature, and quenched by the addition of ethyl acetate,
followed by aqueous sodium sulfate. Solid sodium sulfate was added
to remove excess water. The remaining salts were filtered and
washed with THF. The organic filtrates were combined and
concentrated to give 1.24 g (97%, recovered with 8% progesterone)
white crystalline solid.
##STR00115##
20-S-Hydroxy-progesterone (4)
[0573] A 100 mL RBF was charged with 1.00 g crude compound 5 and
5.00 g manganese dioxide (activated by heating in oven for 2 days
then cooled in a dessicator) and the reactants were suspended in 30
mL chloroform. The mixture was stirred at room temperature
overnight. The mixture was then filtered through a pad of Celite
and rinsed with chloroform. The clear, colorless filtrate was
evaporated to dryness to give an off-white solid. The solid was
recrystallized from ethyl acetate/hexane to give 0.565 g (57%)
white solid.
##STR00116##
20-S--N-Fmoc-L-valine-progesterone (Fmoc 5a)
[0574] An oven dried 50 mL RBF was charged with compound 4 (0.250
g, 0.790 mmol), N-Fmoc-L-valine (0.271 g, 0.798 mmol, 1.01 eq), and
DMAP (0.010 g, 0.079 mmol, 0.100 eq). The flask was sealed,
evacuated, and inert gas flushed and 10 mL anhydrous
dichloromethane was added, followed by addition of 0.869 mL (0.434
mmol, 1.10 eq) 1 M DCC in dichloromethane. The solution was stirred
overnight and then filtered through Celite and washed with
dichloromethane. The crude product was loaded as a silica cake on a
40 g silica column and eluted with a 0-25% ethyl acetate in hexanes
gradient over 45 min. Main product containing fractions were
combined and dried under vacuum to give 0.436 g (87%) white
foam.
##STR00117##
20-S-L-Valine-progesterone (5b)
[0575] Compound 5a (0.374 g, 0.586 mmol) was dissolved in 6 mL
anhydrous acetonitrile in a 25 mL RBF under argon. Piperidine
(0.646 mL, 6.54 mmol, 10.0 eq) was added quickly dropwise at room
temperature. A white clumping precipitate was observed in solution
after 20 minutes. The precipitate was filtered and rinsed with
acetonitrile. The filtrate was concentrated and the resulting white
solid was redissolved in dichloromethane and concentrated in the
presence of 1 g silica. The silica cake was eluted with 0-75% ethyl
acetate in hexanes over 45 minutes on a 12 g silica column. Main
product containing fractions were combined and concentrated to give
a white solid. The solid was recrystallized from hexanes/ethyl
acetate to give 0.097 g (40%) white powdery solid. R.sub.f=0.06
(1:1 EA/hex); .sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.72 (s,
1H), 4.93-4.86 (m, 1H), 3.23 (d, 1H, J=4.4 Hz), 2.46-2.23 (m, 5H),
2.10-0.80 (m, 18H), 1.17 (s, 3H), 1.16 (d, 3H, J=6.4 Hz), 0.98 (d,
3H, J=7.2 Hz), 0.88 (d, 3H, J=6.8 Hz), 0.68 (s, 3H); .sup.13C NMR
(100 MHz, CDCl.sub.3) .delta. 199.8, 175.1, 171.5, 124.0, 73.4,
59.9, 55.4, 55.1, 54.0, 42.5, 39.2, 38.8, 35.9, 35.6, 34.2, 33.0,
32.2 (2C), 25.6, 24.4, 21.1, 20.0, 19.5, 17.6, 17.1, 12.7; IR
(film): 2933, 1721, 1672, 1381, 1187, 1071, 864 cm.sup.-1; HRMS-ESI
m/z 416.3156 ([M+H].sup.+, C.sub.26H.sub.42NO.sub.3 requires
416.3159).
##STR00118##
20-S-L-Valine-progesterone HCl Salt (P1-57)
[0576] Compound 8 (62 mg, 0.150 mmol) was dissolved in 1.5 mL
anhydrous ether in a 5 mL RBF under argon and the solution was
chilled in an ice bath. A 0.158 mL volume (0.158 mmol, 1.05 eq) of
1.00 M hydrochloric acid in diethyl ether was added. A precipitate
formed in solution. The precipitate was filtered and washed with
chilled ether to give 40 mg (59%) off-white solid.
Example 5
Progesterone Prodrug Compounds
[0577] Scheme 15 depicts the synthesis of P1-79, a C20 tryptophan
oxime derivative. Similar reagents and conditions may be used to
prepare C20 oxime derivatives.
##STR00119##
##STR00120##
20-Ketal-progesterone (6)
[0578] Progesterone (25.0 g, 79.5 mmol), oxalic acid (7.16 g, 79.5
mmol, 1.00 eq) and 350 mL benzene were added to a 1 L RBF with stir
bar, followed by 75.4 mL (1.35 mol, 17.0 eq) ethylene glycol. The
flask was fitted with a condenser topped Dean Stark apparatus and
refluxed for 48 h. The solution was cooled and quenched with
saturated sodium bicarbonate solution. The aqueous phase was
extracted with benzene. The organic layers were combined and washed
with DI. The organic layer was treated with magnesium sulfate to
the point of free flowing solid and stirred at room temperature
overnight. The solution was filtered and concentrated to give a
sticky white solid. The solid was recrystallized from petroleum
ether/acetone to give 9.30 g (31% at 94% purity) white solid.
##STR00121##
3-Hydroxy-oxime-20-ketal-progesterone (7a/7b)
[0579] Hydroxylamine HCl (2.78 g, 40.0 mmol, 4.00 eq) was added to
a 100 mL oven dried RBF with 15 mL anhydrous dichloromethane.
Triethylamine (6.97 mL, 50.0 mmol, 5.00 eq) was added and the
mixture was stirred for 45 minutes. Compound 6 was dissolved in 20
mL anhydrous DCM and added quickly dropwise to the reaction
mixture. The reaction was stirred for 24 h at room temp. The
solution was quenched with the addition of DI. The organic layer
was washed with water. The aqueous washes were combined and
extracted with dichloromethane. The organic layers were combined,
dried, filtered, and concentrated with 10 g silica. The silica cake
was eluted with a 0-25% ea in hex gradient over 60 minutes on a 120
g silica column. Main product were recovered as 2.23 g (60%) E
oxime and 1.33 g (36%) Z oxime, both as white solids.
##STR00122##
N-Fmoc-valine-3-E-oxime-20-ketal-progesterone (29)
[0580] Prepared according to the method described for compound 4b.
(99%) clear oil that foamed on drying; R.sub.f=0.54 (1:1 EA/hex);
.sup.1H NIIVIR (600 MHz, CDCl.sub.3) .delta. 7.77 (d, 2H, J 7.8
Hz), 7.61 (dd, 2H, J=7.2, 3.0 Hz), 7.42-7.39 (m, 2H), 7.32 (t, 211,
J 7.6 Hz), 5.98 (s, 1H), 5.46 (d, 1H, J 9.6 Hz), 4.44-4.40 (m, 3H),
4.24 (t, 1H, J=7.2 Hz), 4.02-3.86 (m, 4H), 3.01 (d, 1H, J=16.8),
2.37-0.81 (m, 20H), 1.30 (s, 3H), 1.08 (s, 3H), 1.02 (d, 3H, J 7.2
Hz), 0.99 (d, 3H, J 7.2 Hz), 0.80 (s, 3H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. 170.1, 164.2, 162.4, 156.4, 144.1, 143.9,
127.9, 127.3, 125.3, 120.2, 115.9, 112.0, 67.3, 65.4, 63.4, 58.4,
58.3, 56.0, 53.7, 47.3, 42.0, 39.5, 38.3, 35.3, 34.4, 33.1, 32.1,
31.9, 24.7, 23.9, 23.1, 21.2, 21.0, 19.1, 18.0, 17.8, 13.1; IR
(film): 3347, 2937, 2880, 1756, 1718, 1513, 1374, 1339, 1239, 911,
710 cm.sup.-1; HRMS-ESI m/z 695.4059 ([M+H].sup.+,
C.sub.43H.sub.55N.sub.2O.sub.6 requires 695.4055).
##STR00123##
N-Fmoc-valine-3-E-oxime-progesterone (30)
[0581] Compound 29 (0.265 g, 0.381 mmol) was dissolved in 15 mL
acetone and 0.0164 g (0.0953 mmol, 0.250 eq) PTSA was added. The
reaction was stirred at room temperature for 2 h then heated to
40.degree. C. for 1 h. Ethyl acetate was added and the reaction was
concentrated to remove acetone. Ethyl acetate was added and washed
with water (2.times.10 mL). The aqueous layers were combined and
extracted with ethyl acetate. The organic layers were combined,
washed with brine, dried, and concentrated. The recovered oil was
re-dissolved in DCM and prepared as a silica cake with 0.750 g
silica. The cake was eluted on a 12 g silica column with a 0-35% EA
in hex gradient over 45 minutes. The main product was recovered as
0.245 g (99%) waxy off-white solid. R.sub.f=0.52 (1:1 EA/hex, PMA
stain); .sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 7.77 (d, 2H, J
7.8 Hz), 7.61 (dd, 2H, J 7.2, 2.4 Hz), 7.42-7.39 (m, 2H), 7.32 (t,
2H, J=7.2 Hz), 6.00 (s, 1H), 5.46 (d, 2H, J 9.6 Hz), 4.40-4.40 (m,
3H), 4.24 (t, 1H, J 7.2 Hz), 3.03 (d, 1H, J 17.4 Hz), 2.53 (t, 1H,
J 9.6 Hz), 2.37-0.85 (m, 19H), 2.12 (s, 3H), 1.08 (s, 3H), 1.02 (d,
3H, J 7.2 Hz), 0.99 (d, 3H, J 6.6 Hz), 0.65 (s, 3H); .sup.13C NMR
(150 MHz, CDCl.sub.3) .delta. 209.6, 170.1, 164.1, 161.8, 156.4,
144.1, 143.9, 141.5, 127.9, 127.3, 125.3, 120.2, 116.2, 67.4, 63.7,
58.4, 56.3, 53.6, 47.4, 44.1, 38.9, 38.3, 35.8, 34.4, 32.9, 32.1,
31.9, 31.7, 24.6, 23.0, 21.4, 21.0, 19.1, 18.1, 17.8, 13.5;
HRMS-ESI m/z 651.3799 ([M+H].sup.+, C.sub.41H.sub.51N.sub.2O.sub.5
requires 651.3793).
##STR00124##
3-Valine-E-oxime-progesterone HCl (P1-185)
[0582] Oxime compound 30 (0.260 g, 0.400 mmol) was added to an oven
dried 50 mL RBF and the flask was evacuated and inert gas flushed.
Anhydrous ACN (20 mL) was added and the clear colorless solution
was chilled to 0.degree. C. Freshly distilled piperidine (0.395 mL,
4.00 mmol, 10.0 eq) was added and the solution was stirred and
allowed to gradually equilibrate to room temperature over 30 mm.
The reaction was concentrated with added toluene to give a clear
oil. The oil was loaded neat with minimum DCM rinse onto a 12 g
silica column and eluted with 0-80% ea in hexane over 40 min. Main
product containing fractions were combined and concentrated by
rotary evaporation at 10.degree. C. After being brought to complete
dryness and re-dissolved in 10 mL ethyl acetate, the sample was
concentrated and dried under high vacuum while being chilled in an
ice bath. Anhydrous ether (7-8 mL) was added and the clear
colorless solution was allowed to cool to 0.degree. C. HCl ether
solution (0.195 mL, 2.0 M. 1.0 eq) was added dropwise to the
rapidly stirring solution. A white precipitate was observed. The
mixture was stirred for 15 minutes at 0.degree. C., then filtered
through a 15 mL fine fit glass ground filter and washed with ice
chilled anhydrous ether. (67%, two steps) white solid; R.sub.f=0.25
(1:1 EA/hex, PMA stain); .sup.1H NMR (400 MHz, DMSO) .delta. 8.71
(bs, 2H), 5.90 (s, 1H), 4.00 (bs, 1H), 3.53 (bs, 1H), 3.03 (d, 1H,
J 16.8 Hz), 2.56 (t, 1H, J 9.0 Hz), 2.40-0.79 (m, 20H), 2.06 (s,
3H), 1.05 (s, 3H), 1.02 (d, 3H, J 6.8 Hz), 0.96 (d, 3H, J 6.8 Hz),
0.56 (s, 3H); .sup.13C NMR (150 MHz, CDCl.sub.3) 6209.6, 166.5,
164.8, 162.5, 115.9, 63.7, 58.4, 56.3, 53.6, 44.1, 38.9, 38.3,
35.8, 34.5, 33.0, 32.2, 31.8, 30.3, 24.6, 23.0, 21.5, 19.2, 18.4,
17.9, 15.3, 13.5; IR (solid): 2933, 2874, 2648, 1760, 1699, 1627,
1377, 1358, 1185, 847 cm.sup.-1; HRMS-ESI m/z 429.3109
([M-Cl].sup.+, C.sub.26H.sub.41N.sub.2O.sub.3 requires 429.3112);
Anal. Calcd for C.sub.26H.sub.41ClN.sub.2O.sub.3+1/2H.sub.2O: C,
65.87; H, 8.93; N, 5.91. Found C, 66.03; H, 8.90; N, 5.83.
[0583] The following compounds were prepared according to the
methods developed for the E-oxime pro-drug series:
##STR00125##
N-Fmoc-valine-3-Z-oxime-20-ketal-progesterone (31)
[0584] (99%) white foam; R.sub.f=0.50 (1:1 EA/hex); .sup.1H NMR
(600 MHz, CDCl.sub.3) .delta. 7.77 (d, 2H, J 7.8 Hz), 7.62-7.60 (m,
2H), 7.43-7.36 (m, 2H), 7.32 (t, 2H, J 7.2 Hz), 6.33 (s, 1H), 5.45
(d, 1H, J 9.6 Hz), 4.44 (dd, 1H, J 9.0, 4.8 Hz), 4.40 (d, 2H, J 7.2
Hz), 4.24 (t, 1H, J 7.2 Hz), 4.03-3.84 (m, 4H), 2.60-0.74 (m, 21H),
1.30 (s, 3H), 1.12 (s, 3H), 1.04 (d, 3H, J 6.6 Hz), 1.00 (d, 3H,
J=7.2 Hz), 0.80 (s, 3H); .sup.13C NMR (150 MHz, CDCl.sub.3) .delta.
170.2, 165.7, 161.4, 156.4, 144.1, 144.0, 141.5, 127.9, 127.3,
125.3, 120.2, 112.0, 110.8, 67.3, 65.4, 63.4, 58.4, 58.3, 56.0,
54.0, 47.4, 42.1, 39.5, 39.4, 35.7, 35.3, 34.2, 33.5, 32.6, 31.9,
25.8, 23.9, 23.1, 21.1, 19.3, 18.0, 17.9, 13.1; IR (solid): 3318,
2935, 2876, 1716, 1468, 1371, 1309, 1236, 1042, 862, 739 cm.sup.1
HRMS-ESI m/z 695.4054 ([M+H].sup.+, C.sub.43H.sub.55N.sub.2O.sub.6
requires 695.4066).
##STR00126##
N-Fmoc-valine-3-Z-oxime-progesterone (32)
[0585] (89%) white foam; R.sub.f=0.40 (1:1 EA/hex); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 7.77 (d, 2H, J=7.6 Hz), 7.61 (dd, 2H,
J=7.2, 2.4 Hz), 7.40 (t, 2H, J 7.6 Hz), 7.32 (t, 2H, J 7.6 Hz),
6.35 (s, 1H), 5.47 (d, 1H, J 8.8 Hz), 4.44 (dd, 1H, J 9.2, 4.8 Hz),
4.40 (d, 2H, J 6.8 Hz), 4.24 (t, 1H, J 7.2 Hz), 2.62-0.86 (m, 21H),
2.12 (s, 3H), 1.11 (s, 3H), 1.04 (d, 3H, J 6.8 Hz), 1.00 (d, 3H, J
7.2 Hz), 0.65 (s, 3H); .sup.13C NMR (150 MHz, CDCl.sub.3) .delta.
209.6, 170.2, 165.1, 161.3, 156.4, 144.1, 143.9, 141.5, 127.9,
127.3, 125.3, 120.2, 110.9, 67.3, 63.7, 58.4, 56.2, 53.8, 47.3,
44.1, 39.3, 38.8, 35.7 (2C), 33.3, 32.5, 31.9, 31.7, 24.7, 24.5,
23.0, 21.2, 19.3, 18.0, 17.9, 13.5; IR (solid): 3327, 2935, 1699,
1449, 1371, 1355, 1235, 1032, 859, 760, 740 cm.sup.-1; HRMS-ESI m/z
651.3793 ([M+H].sup.+, C.sub.4, H.sub.51N.sub.2O.sub.5 requires
651.3793).
##STR00127##
3-Valine-Z-oxime-progesterone HCl (P1-186)
[0586] (61%, two steps) white solid; .sup.1H NMR (400 MHz, DMSO)
.delta. 8.74 (bs, 3H), 6.50 (s, 1H), 3.98 (s, 1H), 3.37 (s, 1H),
2.56 (t, 1H, J 8.8 Hz), 2.48-0.82 (m, 20H), 2.06 (s, 3H), 1.09 (s,
3H), 1.01 (d, 3H, J 7.2 Hz), 0.98 (d, 3H, J=6.4 Hz), 0.56 (s, 3H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.6, 167.0, 166.0,
162.0, 111.3, 63.7, 58.2, 56.2, 53.8, 44.1, 39.3, 38.9, 35.7 (2C),
33.3, 32.5, 31.7, 30.3, 24.7, 24.6, 23.0, 21.3, 19.1, 18.5, 18.1,
13.6; IR (solid): 2935, 1756, 1698, 1620, 1379, 1356, 1185, 852
cm.sup.-1 HRMS-ESI m/z 429.3108 ([M-Cl]+,
C.sub.26H.sub.41N.sub.2O.sub.3 requires 429.3112); Anal. Calcd for
C.sub.26H.sub.41ClN.sub.2O.sub.3,+1/2H.sub.2O: C, 65.87; H, 8.93;
N, 5.91. Found C, 65.44; H, 8.94; N, 5.82.
##STR00128##
N-Fmoc-valine-3-hydrazide-20-ketal-progesterone (33)
[0587] Compound 26 (1.79 g, 5.00 mmol) was added to an oven dried
250 mL RBF with 50 mL absolute ethanol. Hydrazine (1.28 mL, 25.0
mmol, 5.00 eq) was added and the reaction was set to reflux
overnight. DMF was added and the solution was concentrated
(40.degree. C. water bath to 25 mbar). Brine was added and the
solution was extracted with EA. The organic layers were combined,
washed with brine, dried, filtered, and concentrated. The crude
mixture (assumed to be 5.00 mmol), was dissolved in anhydrous DCM
and added with N-Fmoc-L-valine (1.87 g, 5.50 mmol, 1.10 eq) and
DMAP (0.0611 g, 0.500 mmol, 0.100 eq) to a 250 mL RBF under argon.
After complete dissolution of the reaction components, DCC (5.50 mL
1 M soln. in DCM, 5.50 mmol, 1.10 eq) was added. After stirring
overnight, the mixture was filtered through Celite. The filtrate
was concentrated and prepared as a silica cake that was eluted on a
120 g silica column with 0-45% EA in hex over 45 min. Main peak
containing fractions were combined and concentrated to give 1.91 g
(55%) off-white solid that was a .about.2:1 mixture of the two main
products. R.sub.f=0.37/0.32 (1:1 EA/hex); major isomer: .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 8.54 (bs, 1H), 7.72-7.16 (m, 8H),
5.80 (s, 1H), 5.65 (d, 1H, J 9.2 Hz), 5.16 (dd, 1H, 9.6, 4.4 Hz),
4.35-3.74 (m, 7H), 2.50-0.64 (m, 21H), 1.30 (s, 3H), 1.06-1.00 (m,
6H), 0.92 (d, 3H, J 6.4 Hz), 0.80 (s, 3H); IR (solid): 3273, 2931,
2874, 1662, 1505, 1240, 1050, 739 cm.sup.-1; HRMS-ESI m/z 716.4032
([M+Na].sup.+, C.sub.43H.sub.55N.sub.3O.sub.5Na requires
716.4045).
##STR00129##
N-Fmoc-valine-3-hydrazide-progesterone (34)
[0588] Compound 33 (0.732 g, 1.05 mmol) was added to a 15 mL 0.85 M
thiourea 1:1 ethanol/water solution. The mixture was heated to
reflux overnight. An orange-amber mass of insoluble oily semi-solid
had formed. The ethanol was removed and DCM and DI were added. The
aqueous layer was extracted with DCM (3.times.50 mL). The organic
layers were combined, washed with brine, dried, filtered, and
concentrated with silica. The silica cake was eluted through a 40 g
silica column with 0-35% ea in hex over 45 minutes. The desired
product(s) were isolated as 0.216 g (32%) off-white solid in a 3:2
mixture of E/Z hydrazides. .sup.1H NMR (400 MHz, CDCl.sub.3)
.delta. 8.63 (bs, 1H), 7.80-7.25 (m, 8H), 5.87 (s, 1H), 5.67 (d,
1H, J 10.0 Hz), 5.16 (dd, 1H, J 9.6, 4.4 Hz), 4.41-4.16 (m, 3H),
2.60-0.78 (m, 21H), 2.13 (s, 3H), 1.04 (s, 3H), 1.01 (d, 3H, J 6.8
Hz), 0.92 (d, 3H, J=6.8 Hz), 0.65 (s, 3H); IR (solid): 3270, 2931,
1698, 1666, 1505, 1234, 1030, 739 cm.sup.-1; HRMS-BSI m/z 650.3944
([M+H].sup.+, C.sub.41H.sub.52N.sub.3O.sub.4 requires
650.3952).
##STR00130##
3-Valine-hydrazide-progesterone (35)
[0589] Compound mixture 34 (0.130 g, 0.200 mmol) was dried in a 25
mL RBF and placed under argon. DMF (5 mL) was added which
completely dissolved the substrate. The solution was chilled in an
ice bath and piperidine (0.206 mL, 2.00 mmol, 10.0 eq) was added.
The ice bath was removed after 15 minutes and the reaction stirred
an additional 15 minutes. Ether was added along with brine. The
aqueous layer was extracted with ether (4.times.20 mL). The organic
layers were combined, washed with brine, dried, filtered, and
concentrated. The sample was loaded in a minimum amount of DCM onto
a 12 g silica column and eluted with 0-5% MeOH in DCM over 35 mm
(initial 5 mm 100% DCM). Main peak containing fractions were
combined and concentrated to give 0.079 g (92%) clear oil. .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 10.20 (bs, 1H), 6.06 (s, 1H),
5.30 (s, 1H), 3.42 (d, 1H, J 3.6 Hz), 2.60-0.78 (m, 22H), 2.11 (s,
3H), 1.06 (s, 3H), 1.01 (d, 3H, J 6.8 Hz), 0.86 (d, 3H, J 6.8 Hz),
0.65 (s, 3H); .sup.13C NMR (100 MHz, CDCl,) .delta. 209.7, 170.0,
156.9, 154.5, 121.5, 63.8, 60.1, 56.3, 53.6, 44.1, 38.9, 37.9,
35.9, 34.9, 32.5, 32.2, 31.7, 30.8, 24.6, 23.0, 21.6, 20.8, 19.8,
17.9, 16.1, 13.5; IR (solid): 3236, 2933, 2874, 1666, 1385, 1358,
1206, 910, 728 cm.sup.-1; HIRMS-BSI m/z 428.3268 ([M+H].sup.+,
C.sub.26H.sub.42N.sub.3O.sub.2 requires 428.3272).
##STR00131##
3-Valine-hydrazide-progesterone (P2-29)
[0590] Prepared according to the method described for compound
P1-31. (43%) off-white solid; .sup.1H NMR (400 MHz, CD.sub.3OD)
.delta. 6.59 (s, 1H), 6.16 (s, 1H), 4.62-4.59 (m, 1H), 3.96 (bs,
2H), 3.02-0.86 (m, 23H), 2.12 (s, 3H) 1.24 (d, 3H, J 7.2 Hz), 1.18
(d, 3H, J 6.8 Hz), 1.11 (s, 3H), 0.67 (s, 3H); HRMSBSI m/z 428.3268
([M+H-2HCl].sup.+, C.sub.26H.sub.42N.sub.3O.sub.2 requires
428.3272); Anal. Calcd for
C.sub.26H.sup.4.sub.3Cl.sub.2N.sub.3O.sub.2+1/2H.sub.2O: C, 61.29;
H, 8.70; N, 8.25. Found C, 61.57; H, 8.85; N, 8.14.
##STR00132##
[0591]
(2R,5S)-3-((E)-((10R,13S,17S)-17-acetyl-10,13-dimethyl-7,8,9,11-tet-
rahydro-1H-cyclopenta[a]phenanthren-3
(2H,6H,10H,12H,13H,14H,15H,16H,17H)-ylidene)amino)-5-isopropyl-2-methylim-
idazolidin-4-one (36). Compound mixture 35 (0.140 g, 0.215 mmol)
was dried in a 25 mL RBF and placed under argon. Acetonitrile was
added (5 mL) along with DMF (2 mL) to completely dissolve the
substrate. The solution was chilled in an ice bath and piperidine
(0.213 mL, 2.15 mmol, 10.0 eq) was added. The ice bath was removed
after 15 minutes and the reaction stirred an additional 15 minutes.
Ethyl acetate was added along with half saturated ammonium chloride
solution. The aqueous layer was extracted with EA and the organic
layers were combined, washed with brine, dried, filtered, and
concentrated. Dichloromethane was added and twice concentrated with
the sample. The resulting pale amber oil was loaded in a minimum
amount of DCM onto a 12 g silica column run in 0-100% ea in hex
over 40 minutes. The two main products were isolated separately as
white crystalline solids (87% combined yield). First eluting
product: R.sub.f=0.32 (95:5 DCM/MeOH); .sup.1H NMR (600 MHz,
CDCl.sub.3) .delta. 5.99 (s, 1H), 4.81 (q, 1H, J 5.4 Hz), 3.48 (d,
1H, J 3.6 Hz), 2.54-2.48 (m, 1H), 2.40-0.80 (m, 21H), 2.11 (s, 3H),
1.32 (d, 3H, J=5.4 Hz), 1.12 (s, 3H), 1.03 (d, 3H, J 6.6 Hz), 0.94
(d, 3H, J 7.2 Hz), 0.65 (s, 3H); .sup.13C NMR (150 MHz, CDCl,)
.delta. 209.6, 170.5, 170.2, 160.8, 120.7. 73.3, 63.8, 63.7, 56.4,
53.8, 44.1, 39.0, 38.5, 35.8 (2C), 32.7, 32.2, 31.8, 31.7, 25.0,
24.6, 23.0, 21.7, 21.3, 19.7, 17.7, 17.4, 13.5; HRMS-ESI m/z
454.3430 ([M+H].sup.+, C.sub.28H.sub.44N.sub.3O.sub.2 requires
454.3428).
TABLE-US-00004 TABLE 3 Crystal data and structure refinement for
compound 36. Identification code p2264s Empirical Formula C28 H42
N3 O2 Formula weight 452.65 Temperature 173(2) K Wavelength 1.54178
.ANG. Crystal system Monoclinic Space group C2 Unit cell dimensions
a = 65.281(5) .ANG. .alpha. = 90.degree.. b = 5.9668(6) .ANG.
.beta. = 96.847(6).degree.. c = 10.6916(8) .ANG. .gamma. =
90.degree.. Volume 4134.8(6) .ANG..sup.3 Z 4 Density (calculated)
0.727 Mg/m.sup.3 Absorption coefficient 0.355 mm.sup.-1 F(000) 988
Crystal size 0.42 .times. 0.07 .times. 0.03 mm.sup.3 Theta range
for data 2.73 to 66.48.degree.. collection Index ranges -76 <= h
<= 73, -6 <= k <= 6, -12 <= 1 <= 12 Reflections
collected 12330 Independent reflections 5937 [R(int) = 0.0866]
Completeness to 96.0% theta = 66.48.degree. Absorption correction
Semi-empirical from equivalents Max. and min. 0.9894 and 0.8651
transmission Refinement method Full-matrix least-squares on F.sup.2
Data/restraints/ 5937/1/306 parameters Goodness-of-fit on F.sup.2
1.061 Final R indices R1 - 0.1571, [I > 2sigma(I)] wR2 - 0.3866
R indices (all data) R1 = 0.2305, wR2 = 0.4236 Absolute structure
0.6(14) parameter Extinction coefficient 0.0037(6) Largest diff
peak and 0.470 and -0.465 e. .ANG.-.sup.3 hole
##STR00133##
O--N-Fmoc-L-tryptophan-C3-oxime-C20-ketal-progesterone (8a)
[0592] An oven dried 25 mL RBF was charged with oxime 11 (0.187 g,
0.500 mmol), N-Fmoc-L-tryptophan (0.242 g, 0.22 mmol, 1.05 eq), and
DMAP (0.0061 g, 0.021 mmol, 0.10 eq). The flask was sealed,
evacuated, and inert gas flushed and 15 mL anhydrous
dichloromethane was added, followed after complete dissolution by
addition of 0.550 mL (0.23 mmol, 1.10 eq) 1 M DCC in
dichloromethane. The solution was stirred for 16 h at room
temperature. The mixture was filtered through Celite, the filtrates
concentrated, and the crude oil loaded as a silica cake onto 1.17 g
silica. The cake was eluted on a 40 g silica column in 0-35% ea in
hex over 90 minutes. The main product peak was isolated as 0.383 g
(98%) white foam.
##STR00134##
O--N-Fmoc-L-tryptophan-C3-oxime-progesterone (Fmoc 9a)
[0593] Compound 8a (0.350 g, 0.448 mmol) was dissolved in 15 mL
acetone and 0.0193 g (0.112 mmol, 0.250 eq) PTSA was added. The
reaction was stirred at room temperature for 2.5 h. Ethyl acetate
was added and the solvent was concentrated twice with re-addition
of ethyl acetate. The ethyl acetate was washed with water
(2.times.25 mL). The aqueous layers were combined and extracted
with ethyl acetate. The organic layers were combined, washed with
brine, dried, and concentrated to give a yellow oil that solidified
on further drying. The mixture was redissolved in a minimum amount
of DCM with toluene and loaded neat onto a 40 g silica column and
eluted in a 0-40% ea in hex gradient over 70 minutes to give 0.315
g (95%) pale amber foam.
##STR00135##
O-L-Tryptophan-C3-oxime-progesterone (9a)
[0594] Compound Fmoc 9a (0.280 g, 0.379 mmol) was added to an oven
dried 25 mL RBF. Anhydrous acetonitrile (7.5 mL) was added,
followed by piperidine (0.141 mL, 1.42 mmol, 10.0 eq). The reaction
was stirred for 30 minutes at room temperature. The solvent was
removed by evaporation and the crude oil redissolved in toluene and
reduced to dryness twice in succession. The crude off-white solid
was redissolved in a minimum amount of DCM, loaded neat onto a 12 g
silica column, and eluted with a 0-95% ea in hex gradient over 60
min. The main product was obtained as 0.120 g (61%) off-white
solid.
##STR00136##
O-L-Tryptophan-C3-oxime-progesterone HCl Salt (P1-79)
[0595] Compound 14 (46 mg, 0.089 mmol) was dissolved in 2.5 mL
anhydrous ether in a 10 mL RBF under argon. The solution was
chilled in an ice bath and 0.195 mL 1 M HCl solution in ether was
added. A white precipitate was observed to have immediately formed
in solution. The mixture was filtered and the precipitate was
washed with cold ether to give 21 mg (43%) white solid.
##STR00137##
3-Hydrazine-20-ketal-progesterone (10)
[0596] Compound 6 (0.377 g, 95% w/w, 1.00 mmol) was added to an
oven dried 25 mL RBF and 5 mL absolute ethanol was added. Hydrazine
(5.00 mL 1.0 M solution in THF, 5.00 mmol, 5.00 eq) was added which
served to completely dissolve the starting material. This was
stirred at room temperature for 1.5 h and set to reflux overnight.
The solution was concentrated and dried under vacuum to give a
white foam. Dichloromethane was added and the solution was
re-concentrated to generate a solid that was filtered and washed
with 3:1 hex/ether to give 0.164 g (44%) pale yellow crystals.
Example 6
Allopregnanolone Derivatives
[0597] Scheme 16 below shows the synthesis of C3 ALLO valine
derivatives. Other C3 amino acid ALLO derivatives may be prepared
with similar reagents and conditions.
##STR00138##
##STR00139##
3.beta.-hydroxy-5.alpha.-pregnan-20-one (12)
[0598] An oven dried 500 mL RBF was charged with 10% palladium on
carbon (0.400 g) and 5-pregnen-3-beta-ol-20-one (4.00 g, 12.6 mmol)
and the flask was evacuated and flushed with argon. A 200 mL volume
of absolute ethanol was added and the flask was flushed with
hydrogen. The reaction was stirred at room temperature for 4 h. The
mixture was filtered through Celite and the recovered clear,
colorless filtrate was concentrated to reveal a white solid of mass
4.08 g. The solid was recrystallized from hexane/ethyl acetate
(.about.3:1 total 175 mL) to give 3.19 g white solid. A second
recrystallization provided an additional 0.43 g for a total of 3.62
g (90%) white crystalline solid.
##STR00140##
3.beta.-N-Fmoc-L-valine-5.alpha.-pregnan-20-one (Fmoc 13a)
[0599] An oven dried 25 mL RBF was charged with compound 20 (0.318
g, 1.00 mmol), N-Fmoc-L-valine (0.356 g, 1.05 mmol, 1.05 eq) and
DMAP (12 mg, 0.100 mmol, 0.10 eq). The flask was sealed, evacuated
and inert gas flushed, and 9 mL anhydrous DCM was added, followed
after complete substrate dissolution by 1.10 mL (1.10 mmol, 1.10
eq) 1.0 M DCC in DCM. The reaction mixture was stirred at room
temperature for 24 h. The mixture was filtered through Celite and
rinsed with DCM. The sample was prepared as a silica cake and
eluted on a 40 g silica column with 0-25% ea in hex over 45 min.
Main product containing fractions were combined and isolated as
0.578 g (90%) white foam.
##STR00141##
3.beta.-L-valine-5.alpha.-pregnan-20-one (13a)
[0600] A 25 mL RBF was charged with compound Fmoc 13a (0.488 g,
0.725 mmol) and 7.5 mL acetonitrile. Piperidine (0.716 mL, 7.25
mmol, 10.0 eq) was added and the solution was stirred at room
temperature for 30 min. Toluene was added and the solution was
concentrated 3 times with addition of toluene. The resulting white
solid was redissolved in a minimum amount of toluene and loaded
onto a 12 g silica column. The column was eluted with 0-100% ea in
hex over 40 minutes. The main product was obtained as 0.317 g (99%)
clear/white semi-solid.
##STR00142##
3.beta.-L-valine-5.alpha.-pregnan-20-one HCl Salt (P1-123)
[0601] Compound 22 (0.317 g, 0.759 mmol) was dissolved in
.about.2:1 anhydrous ether/DCM (6 mL total) under argon. The clear,
slightly amber solution was chilled in an ice bath and 0.759 mL
(0.759 mmol, 1.0 eq) 1 M HCl in ether solution was added slowly
dropwise. A white precipitate was observed in solution. The
solution was stirred at 0.degree. C. for 30 min and then filtered.
The precipitate was washed with ice chilled ether. The product was
recovered as a slightly off-white solid of mass 0.175 g (51%).
##STR00143##
3.alpha.-hydroxy-5.alpha.-pregnan-20-one (14)
[0602] An oven dried 100 mL RBF with magnetic stir bar was charged
with 1.59 g (5.00 mmol) compound 20 and 15 mL anhydrous THF.
Diethylazodicarboxylate (2.85 mL 40% soln. in toluene, 6.25 mmol,
1.25 eq) was added, followed by trifluoroacetic acid (0.482 mL,
6.25 mmol, 1.25 eq) and the flask was set in a room temperature
water bath. To this pale amber suspension was added
triphenylphosphine (1.64 g, 6.25 mmol, 1.25 eq). Sodium benzoate
(0.901 g, 6.25 mmol, 1.25 eq) was then added and the suspension was
stirred under argon for 24 h at room temperature. The THF was
completely removed with methanol addition/evaporation. Methanol (20
mL) was then added. The flask was fitted with a drying tube topped
condenser and set for reflux. After 24 h, the methanol was removed
and the remaining solid was redissolved in DCM. The organic layer
was washed with DI (3.times.20 mL). The aqueous layers were
combined and extracted with DCM. The organic layers were combined,
dried, filtered, and concentrated to give a white solid. The solid
was prepared as a silica cake and eluted with 0-35% ea in hex on a
120 g silica column over 40 min. Main product containing fractions
were combined and concentrated to give 1.46 g (92%) white
solid.
##STR00144##
3.alpha.-N-Fmoc-L-valine-5.alpha.-pregnan-20-one (Fmoc 15a)
[0603] An oven dried 50 mL RBF was charged with compound 14 (0.478
g, 1.50 mmol), N-Fmoc-L-valine (0.535 g, 1.58 mmol, 1.05 eq), and
DMAP (18 mg, 0.150 mmol, 0.10 eq). The flask was sealed, evacuated
and inert gas flushed, and 15 mL anhydrous DCM was added, followed
after complete substrate dissolution by 1.65 mL (1.65 mmol, 1.10
eq) 1.0 M DCC in DCM. The flask was stirred at room temperature for
24 h. The mixture was filtered through Celite and rinsed with DCM.
Silica (.about.3 g) was added and the mixture was concentrated. The
silica cake was eluted on a 40 g silica column with 0-25% ea in hex
over 45 min. The main product was isolated as 0.834 g (87%) white
foam.
##STR00145##
3.alpha.-L-Valine-5.alpha.-pregnan-20-one (15a)
[0604] A 25 mL RBF was charged with compound Fmoc 15a (0.320 g,
0.500 mmol), 5 mL ACN, and 3 mL DMF. Piperidine (0.494 mL, 5.00
mmol, 10.0 eq) was added. The solution was stirred at room
temperature for 30 minutes. Toluene was added and the solution was
concentrated 3 times with addition of toluene. The pale amber oil
was loaded in a minimum amount of toluene onto a 12 g silica
column. The column was eluted with 0-100% ea in hex over 40
minutes. Main product fractions were combined to give 0.196 g (94%)
sticky white solid.
##STR00146##
3.alpha.-L-Valine-5.alpha.-pregnan-20-one HCl Salt (P1-131)
[0605] Compound 25 (0.251 g, 0.600 mmol) was dissolved in 6 mL
anhydrous ether under argon. The clear solution was chilled in an
ice bath and 0.300 mL (0.600 mmol, 1.0 eq) 2.0 M HCl/ether solution
was added slowly dropwise. A white precipitate was observed in
solution. The solution was stirred at 0.degree. C. for 30 min and
then filtered. The precipitate was washed with ice chilled ether.
The product was recovered as 0.150 g (55%) slightly off-white
solid.
[0606] Scheme 17 below shows the synthesis of 5.beta. ALLO isomers
17a and 17b and C3 valine derivatives of the isomers. The isomers
17a and 17b were separated by conventional chromatography and
carried forward as described above to prepare the valine
derivatives P1-133 and P1-135. Derivatives with other amino acids
may be prepared from compounds 17a and 17b using similar reagents
and conditions.
##STR00147##
##STR00148##
5.beta.-Pregnane-3,20-dione (16)
[0607] A three necked 500 mL RBF was charged with progesterone
(2.00 g, 6.36 mmol), 5% Pd/CaCO.sub.3 (0.180 g, 9% w/w), 200 mL
absolute ethanol, and KOH (0.360 g in 1 mL DI). The flask was
evacuated and flushed with hydrogen and the reaction stirred for 1
h. The ethanol was removed and the residue was redissolved in ether
and washed with water. The water layer was extracted with ether
(2.times.50 mL). The aqueous layer was then acidified to pH<3
with 1 M HCl and extracted with ether. The organic layers were
combined, dried, filtered, and concentrated to give an off-white
solid of mass 2.08 g. The sample was loaded in a minimum amount of
toluene onto a 120 g silica column and eluted with 0-35% ea in hex
gradient. The main product was recovered as 1.20 g (60%) white
solid.
##STR00149##
3-Hydroxy-5.beta.-pregnane-20-one (17a/17b)
[0608] A 250 mL RBF was charged with compound 26 (1.00 g, 3.16
mmol) and 40 mL absolute ethanol. The solution was warmed in an oil
bath to 50.degree. C. and sodium borohydride (0.179 g, 4.74 mmol,
1.50 eq) was added. The reaction was stirred for 10 min and 75-100
mL hot water was added until a slight cloudiness remained in
solution. The solution was then allowed to cool gradually to room
temperature and chilled in a 4.degree. C. freezer for 3 h. The
mixture was filtered and the white solid was washed with 30%
ethanol in DI. After drying, the recovered solids were loaded in a
minimum amount of DCM onto a 120 g silica column and eluted with
0-25% ea/hex over 60 min. Main product containing fractions were
combined and concentrated to give 0.710 g (71%)
3.alpha.-hydroxy-5.beta.-pregnane-20-one and 0.110 g (11%)
3.beta.-hydroxy-5.beta.-pregnane-20-one isomer.
##STR00150##
3.alpha.-N-Fmoc-L-valine-5.beta.-pregnane-20-one (Fmoc 18a)
[0609] An oven dried 25 mL RBF was charged with compound 27 (0.333
g, 1.05 mmol), N-Fmoc-L-valine-OH (0.373 g, 1.10 mmol, 1.05 eq),
and DMAP (12.8 mg, 0.10 mmol, 0.10 eq). The flask was sealed,
evacuated and inert gas flushed and 9 mL anhydrous DCM was added,
followed after complete substrate dissolution by 1.15 mL (1.15
mmol, 1.10 eq) 1.0 M DCC in DCM. A white precipitate appeared in
solution during DCC addition. The flask was stirred at room
temperature for 24 h. The mixture was filtered through Celite and
rinsed with DCM. The filtrate was concentrated with 2 g silica and
the silica cake was eluted on a 40 g silica column with 0-25% ea in
hex over 45 min. The main product was recovered as 0.541 g (81%)
white foam.
##STR00151##
3.alpha.-L-valine-5.beta.-pregnane-20-one (18a)
[0610] A 25 mL RBF was charged with compound Fmoc 18a (0.500 g,
0.742 mmol) and 7 mL ACN. Piperidine (0.733 mL, 7.42 mmol, 10.0 eq)
was added and the solution was stirred at room temperature for 30
minutes. A flaky white precipitate appeared in solution. The
precipitate was filtered and washed with ACN. The organic layers
were combined with toluene and the solution was concentrated 3
times with addition of toluene. The white solid was redissolved in
a minimum amount of toluene and loaded onto a 12 g silica column.
The column was eluted with 0-75% ea in hex over 40 minutes. The
main product was isolated as 0.301 g (97%) white solid.
##STR00152##
3.alpha.-L-Valine-5.beta.-pregnane-20-one HCl Salt (P1-133)
[0611] Compound 30 (0.155 g, 0.371 mmol) was dissolved in 4 mL
anhydrous ether under argon. The clear solution was chilled in an
ice bath and 0.186 mL (0.371 mmol, 1.0 eq) 2 M HCl in ether
solution was added slowly dropwise. A white precipitate was
observed in solution. The solution was stirred at 0.degree. C. for
30 minutes and then filtered. The precipitate was washed with ice
chilled 2:1 hex/ether. The product was recovered as a slightly
off-white solid of mass 0.120 g (71%).
##STR00153##
3.beta.-N-Fmoc-L-valine-5.beta.-pregnane-20-one (Fmoc 18b)
[0612] An oven dried 25 mL RBF was charged with compound 28 (0.234
g, 0.735 mmol), N-Fmoc-L-valine (0.262 g, 1.10 mmol, 1.05 eq) and
DMAP (9 mg, 0.10 mmol, 0.10 eq). The flask was sealed, evacuated
and inert gas flushed, and 8 mL anhydrous DCM was added, followed
after complete substrate dissolution by 0.808 mL (0.808 mmol, 1.10
eq) 1.0 M DCC in DCM. The flask was stirred at room temperature for
24 h. The mixture was concentrated with 1.5 g silica, and the
silica cake was eluted on a 40 g silica column with 0-25% ea in hex
over 45 min. The main product was isolated as 0.345 g (73%) white
foam.
##STR00154##
3.beta.-L-Valine-5.beta.-pregnane-20-one (18b)
[0613] A 25 mL RBF was charged with compound 31 (0.307 g, 0.456
mmol) and dissolved in 7 mL ACN. Piperidine (0.450 mL, 4.56 mmol,
10.0 eq) was added and the solution was stirred at room temperature
for 15 minutes. The precipitate was filtered and washed with ACN.
The organic layers were combined with toluene and the solution was
concentrated 3 times. The white solid was redissolved in a minimum
amount of toluene, loaded onto a 12 g silica column, and eluted
with 0-75% ea in hex over 35 minutes. The product was obtained as
0.176 g (93%) white foam.
##STR00155##
3.beta.-L-Valine-5.beta.-pregnane-20-one HCl Salt (P1-135)
[0614] Compound 32 (0.123 g, 0.290 mmol) was dissolved in 3 mL
anhydrous ether under argon. The clear solution was chilled in an
ice bath and 0.15 mL (0.29 mmol, 1.0 eq) 2 M HCl in ether solution
was added slowly dropwise. A white precipitate was observed in
solution. The solution was stirred at 0.degree. C. for 30 minutes
and then filtered. The precipitate was washed with ice chilled 2:1
hex/ether. The product was recovered as a slightly off-white solid
of mass 0.052 g (39%).
Enantiomeric Progesterone Synthesis
[0615] The following compounds were prepared according to
previously published methods (Micheli et al., J. Org. Chem., 1975,
40, 675-681; Hajos et al., J. Org. Chem., 1973, 38, 3239-3243;
Hajos et al., J. Org. Chem., 1973, 38, 3244-3249; Rychnovsky et al.
J. Org. Chem., 1992, 57, 2732-2736).
##STR00156##
2-Methyl-2-(3-oxobutyl)-1,3-cyclopentanedione (38)
[0616] (95%) amber oil; R.sup.f=0.62 (95:5 DCM/MeOH); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 2.87-2.69 (m, 4H), 2.43 (t, 2H, J
7.2), 2.08 (s, 3H), 1.86 (t, 2H, J=7.2), 1.09 (s, 3H); .sup.13C NMR
(100 MHz, CDCl.sub.3) .delta. 216.0 (2C), 208.1, 55.3, 37.6, 34.9
(2C), 30.2, 27.9, 19.2; JR (film): 2930, 1712, 1366, 1169; HRMS-ESI
m/z 183.1014 ([M+H].sup.+C.sub.10H.sub.15O.sub.3 requires
183.1016).
##STR00157##
(R)-7a-Methyl-2,3,7,7a-tetrahydro-6H-indene-1,5-dione (39)
[0617] (80%) pale tan solid; R.sub.f=0.47 (95:5 DCM/MeOH); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 5.95 (d, 1H, J=2.4), 3.00-2.90
(m, 1H), 2.82-2.70 (m, 2H), 2.56-2.37 (m, 3H), 2.09 (ddd, 1H,
J=13.6, 2.4, 2.0), 1.83 (dt, 1H, J=136, 5.2), 1.30 (s, 3H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 216.8, 198.4, 170.0,
124.0, 48.9, 36.0, 33.0, 29.3, 27.0, 20.7; JR (solid): 2970, 2876,
1742, 1699, 1660, 1447, 1146 cm.sup.1; HRMS-ESI m/z 165.0911
([M+H].sup.+, C.sub.10H.sub.13O.sub.2 requires 165.0910).
##STR00158##
(1R,7aR)-1-Hydroxy-7a-methyl-1,2,3,6,7,7a-hexahydro-inden-5-one
(40)
[0618] (94%) amber semisolid; R.sub.f=0.17 (95:5 DCM/MeOH); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 5.78 (s, 1H), 3.84 (t, 1H, J=8.8
Hz), 2.75-2.64 (m, 1H), 2.57-2.33 (m, 4H), 2.17-2.07 (m, 2H),
1.88-1.72 (m, 2H), 1.14 (s, 3H); .sup.3C NMR (100 MHz, CDCl.sub.3)
.delta. 199.7, 175.6, 123.5, 80.7, 45.4, 34.2, 33.5, 29.2, 26.6,
15.3; IR (film): 3335 (br), 2935, 1632, 1326, 1221, 1075 cm.sup.-1;
HRMS-ESI m/z 167.1068 ([M+H].sup.+, C.sub.10H.sub.15O.sub.2
requires 167.1067).
##STR00159##
(1R,7aR)-1-tert-Butoxy-7a-methyl-1,2,3,6,7,7a-hexahydro-inden-5-one
(41)
[0619] (84%) pale yellow solid; R.sub.f=0.48 (95:5 DCM/MeOH);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.75 (s, 1H), 3.56 (dd,
1H, J=9.6, 7.6 Hz), 2.72-2.63 (m, 1H), 2.55-2.47 (m, 1H), 2.46-2.31
(m, 2H), 2.05-1.93 (m, 2H), 1.84-1.67 (m, 2H), 1.17 (s, 9H), 1.10
(s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 199.7, 175.8,
123.1, 79.8, 73.2, 45.0, 34.5, 33.6, 29.7, 28.8 (3C), 27.0, 15.9;
IR (solid): 2972, 1669, 1361, 1198, 1089 cm.sup.-1; HRMS-ESI m/z
223.1691 ([M+H].sup.+, C.sub.14H.sub.23O.sub.2 requires
223.1693).
##STR00160##
(-)-(1R,7aR)-5,6,7,7a-Tetrahydro-1-tert-butoxy-7a-methyl-5-oxo-4-indancar-
boxylic acid (43)
[0620] (71%) brown-yellow solid; R.sub.f=0.21 (DCM/EA);
[.alpha.].sup.23.sub.D-31.1 (c=1.00, CHCl.sub.3) [lit. -37 (c=1.02
CHCl.sub.3)].sup.129 1H NMR (400 MHz, CDCl.sub.3) .delta. 3.67 (dd,
1H, J=10.2, 7.2 Hz), 3.33-3.15 (m, 2H), 2.84-2.61 (m, 2H),
2.12-2.03 (m, 2H), 1.93-1.77 (m, 2H), 1.19 (s, 12H); .sup.13C NMR
(100 MHz, CDCl.sub.3) .delta. 203.1, 196.5, 164.5, 120.5, 78.9,
73.7, 48.5, 33.7, 32.1, 31.6, 30.1, 28.8 (3C), 16.5; IR (solid):
2967, 2750, 1732, 1625, 1599, 1437, 1190, 1099 cm.sup.-1; HRMS-ESI
m/z 267.1588 ([M+H].sup.+, C.sub.15H.sub.23O.sub.4 requires
267.1591).
##STR00161##
(-)-(1R,3aS,7aR)-1-tert-Butoxy-7a-methyl-3a,6,7,7a,-tetrahydro-4-methylen-
eindan-5(4H)-one (45)
[0621] (79%) brown oil; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
5.93 (q, 1H, J 2.8, 1.6 Hz), 5.01 (q, 1H, J 2.4, 1.6 Hz), 3.64-3.57
(m, 1H), 2.59-1.48 (m, 9H), 1.16 (s, 9H), 0.78 (s, 3H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 202.5, 147.5, 118.3, 80.2, 72.8,
49.1, 43.3, 36.2, 34.1, 31.9, 28.9 (3C), 22.9, 11.4; IR (film):
2970, 2874, 1694, 1361, 1191, 1090 cm.sup.-1; HRMS-ESI m/z 237.1847
([M+H].sup.+, C.sub.15H.sub.25O.sub.2 requires 237.1849).
##STR00162##
5-[(1,3-Dioxolan-2-yl)ethyl]hexanoate (47)
[0622] (93%) pale amber oil; R.sub.f=0.40 (2:1 hex/EA, PMA stain);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 4.12 (q, 2H, J 7.2 Hz),
3.96-3.91 (m, 4H), 2.32 (t, 2H, J=7.2 Hz), 1.78-1.64 (m, 4H), 1.32
(s, 3H), 1.25 (t, 3H, J=7.2 Hz); .sup.13C NMR (100 MHz, CDCl.sub.3)
.delta. 173.7, 109.9, 64.8 (2C), 60.4, 38.5, 34.5, 24.0, 19.8,
14.4; IR (neat): 2981, 2880, 1731, 1375, 1176, 1050, 855 cm.sup.-1;
HRMS-ESI m/z 203.11277 ([M+H].sup.+, C.sub.10H,.sub.90.sub.4
requires 203.1278).
##STR00163##
5-(1,3-Dioxolan-2-yI)hexanol (48)
[0623] (99%) clear oil; R.sub.f=0.14 (1:1 hex/EA, PMA stain);
.sup.1H NMR (600 MHz, CDCl.sub.3) .delta. 3.97-3.91 (m, 4H), 3.65
(t, 2H, J 6.6 Hz), 1.69-1.66 (m, 2H), 1.61-1.56 (m, 3H), 1.50-1.45
(m, 2H), 1.32 (s, 3H); .sup.13C NMR (150 MHz, CDCl.sub.3) .delta.
110.2, 64.8 (2C), 63.0, 39.0, 33.0, 24.0, 20.4; IR (neat): 3417
(br), 2942, 2873, 1376, 1220, 1139, 1037 cm.sup.-1; HRMS-ESI m/z
160.13330 ([M-OH+NH.sub.3].sup.+, C.sub.8H.sub.18N0.sub.2 requires
160.1332).
##STR00164##
5-(1,3-Dioxolan-2-yI)hexanal (49)
[0624] (81%) amber oil; R.sub.f=0.37 (1:1 hex/EA, PMA stain);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 9.75 (t, 1H, J 1.6 Hz),
3.97-3.88 (m, 4H), 2.46 (dt, 2H, J 7.2, 1.6 Hz), 1.79-1.64 (m, 4H),
1.31 (s, 3H); .sup.13C NMR (100 MHz, CDCl,) .delta. 202.6, 109.8,
64.8 (2C), 44.0, 38.4, 24.0, 16.8; IR (film): 2982, 2881, 1721,
1376, 1211, 1068, 852 cm.sup.-1; HRMS-ESI m/z 159.1014
([M+H].sup.+, C.sub.8H.sub.15O.sub.3 requires 159.1016).
##STR00165##
Ethyl 7-(1,3-Dioxolan-2-yI)-3-oxooctanoate (50)
[0625] (78%) pale green-amber oil; .sup.1H NMR (600 MHz,
CDCl.sub.3) .delta. 4.19 (q, 2H, J 7.2 Hz), 3.95-3.91 (m, 4H), 3.43
(s, 2H), 2.58 (t, 2H, J 7.2 Hz), 1.74-1.63 (m, 4H), 1.31 (s, 3H),
1.28 (t, 3H, J 7.2 Hz); `.sup.13C NMIR (150 MHz, CDCl.sub.3)
.delta. 202.8, 167.5, 110.0, 64.8 (2C), 61.5, 49.5, 43.0, 38.2,
23S, 18.2, 14.3 cm.sup.-1.
##STR00166##
(-)-3.alpha.-tert-Butoxy-3a.alpha.-methyl-1,2,3,3a,4,5,8,9,9a.quadrature.-
,9ba-dodecahydro-6-[2-(2-methyl-1,3-dioxolan-2-yI)-ethyl]-7H-benz[e]inden--
7-one (51)
[0626] (72%) brown oil; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
3.98-3.91 (m, 4H), 3.40 (t, 1H, J=8.4 Hz), 2.84-2.75 (m, 1H),
2.50-1.04 (m, 17H), 1.14 (s, 9H), 0.88 (s, 3H); .sup.13C NMR (100
MHz, CDCl,) .delta. 199.2, 160.2, 134.1, 110.0, 80.4, 72.7, 64.8
(2C), 51.0, 42.2, 39.1, 38.2, 37.2, 36.8, 31.3, 28.9 (3C), 26.9,
26.8, 24.1, 23.7, 20.3, 11.1; IR (film): 2970, 2872, 1663, 1361,
1194, 1095, 1058 cm.sup.-1; HRMS-ESI m/z 391.2837 ([M+H].sup.+,
C.sub.24H.sub.39O.sub.4 requires 391.2843).
##STR00167##
(-)-3.quadrature.-ter-Butoxy-3a.quadrature.,5a.quadrature.-dimethyl-1,2,3-
,3a,4,5,5a,6,8,9,9a.quadrature.,9a-dodecahydro-6-[2-(2-methyl-1,3-dioxolan-
-2-yl)-ethyl]-7H-benz[c]inden-7-one (52)
[0627] (53%) brown oil; .sup.1H NMR (400 MHz, CDCl,) .delta.
3.98-3.91 (m, 4H), 3.37 (1, 1H, J=8.4 Hz), 2.61-2.40 (m, 1H),
2.28-2.19 (m, 1H), 1.96-0.90 (m, 17H), 1.36 (s, 3H), 1.13 (s, 9H),
1.10 (s, 3H), 0.78 (s, 3H); .sup.13C NMIR (100 MHz, CDCl.sub.3)
.delta. 215.1, 110.5, 80.8, 72.5, 64.7, 64.7, 50.8, 50.6, 47.8,
42.7, 38.3, 36.9, 35.0, 33.2, 31.2, 31.0, 29.2, 28.9 (3C), 24.0,
23.6, 21.4, 21.1, 11.8; IR (film): 2967, 2937, 1703, 1360, 1195,
1067, 1040, 948 cm.sup.-1; HRMS-ESI m/z 407.3154 ([M+H]+,
C.sub.25H.sub.43O.sub.4 requires 407.3156).
##STR00168##
ent-Testosterone (53)
[0628] (71%) white solid; R.sub.f=0.22 (1:1 EA/hex); .sup.1H NMIR
(400 MHz, CDCl.sub.3) .delta. 5.73 (s, 1H), 3.68-3.62 (m, 1H),
2.48-2.23 (m, 4H), 2.13-1.98 (m, 2H), 1.89-1.23 (m, 10H), 1.19 (s,
3H), 1.13-0.88 (m, 4H), 0.79 (s, 3H); .sup.13C NMR (150 MHz,
CDCl.sub.3) .delta. 199.8, 171.5, 124.1, 81.8, 54.1, 50.7, 43.0,
38.9, 36.6, 35.9, 35.8, 34.1, 33.0, 31.7, 30.6, 23.5, 20.8, 17.6,
11.2; IR (solid): 3391, 2930, 2880, 1645, 1612, 1229, 1056
cm.sup.-1; HRMS-ESI m/z 289.2158 ([M+H]+, C.sub.19H.sub.29O.sub.2
requires 289.2162).
[0629] The following compounds were prepared based on previously
published methods (Auchus et al. Arch. Biochem. Biophys. 2003, 409,
134-144).
##STR00169##
ent-(17.beta.)-17-Hydroxyandrost-5-en-3-one cyclic
3-(1,2-ethanediyl acetal) (54)
[0630] (71%) white solid; R.sub.f=0.42 (1:1 EA/hex); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 5.34-5.30 (m, 1H), 3.96-3.89 (m, 4H),
3.62 (t, 1H, J 8.4 Hz), 2.55 (dq, 1H, J 14.2, 3.0 Hz), 2.12-1.90
(m, 3H), 1.83-0.88 (m, 16H), 1.01 (s, 3H), 0.74 (s, 3H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 140.4, 122.1, 109.6, 82.0, 64.6,
64.4, 51.5, 50.0, 42.9, 42.0, 36.9, 36.8, 36.5, 32.1, 31.5, 31.2,
30.7, 23.7, 20.8, 19.1, 11.2; IR (solid): 3213, 2933, 2886, 1093,
1058 cm.sup.-1; HRMS-ESI m/z 333.2426 ([M+H]+,
C.sub.21H.sub.33O.sub.3 requires 333.2424).
##STR00170##
ent-Androst-5-ene-3,17-dione cyclic 3-(1,2-ethanediyl acetal)
(55)
[0631] (95%) white solid; R.sub.f=0.52 (1:1 EA/hex); .sup.1H NMR
(400 MHz, CDCl.sub.3) .delta. 5.41-5.36 (m, 1H), 3.99-3.92 (m, 4H),
2.59 (dq, 1H, J=14.0, 2.8 Hz), 2.46 (dd, 1H, J=19.2, 8.0 Hz),
2.17-2.03 (m, 3H), 1.99-1.90 (m, 1H), 1.88-1.08 (m, 13H), 1.06 (s,
3H), 0.89 (s, 3H); `.sup.13C NMR (100 MHz, CDCl,) .delta. 221.4,
140.6, 121.6, 109.5, 64.7, 64.5, 51.9, 50.0, 47.8, 42.0, 36.9,
36.4, 36.0, 31.7, 31.6, 31.2, 30.8, 22.1, 20.5, 19.1, 13.8; IR
(solid): 2946, 2933, 2883, 1735, 1373, 1092, 992 cm.sup.-1;
HRMS-ESI m/z 331.2283 ([M+H].sup.+, C.sub.21H.sub.31O.sub.3
requires 331.2268).
##STR00171##
ent-[(172)-Pregna-5,17(20)-dien-3-one cyclic (1,2-ethanediyl
acetal)] (56)
[0632] (76%) white solid; R.sub.f=0.73 (1:1 BA/hex, KMnO4 stain);
.sup.1H NMR (400 MHz, CDCl,) .delta. 5.38-5.36 (m, 1H), 5.17-5.10
(m, 1H), 4.00-3.92 (m, 4H), 2.61-2.54 (m, 1H), 2.42-1.07 (m, 21H),
1.05 (s, 3H), 0.90 (s, 3H); .sup.13C NMR (100 MHz, CDCl,) .delta.
150.5, 140.4, 122.3, 113.7, 109.7, 64.7, 64.4, 56.6, 49.9, 44.3,
42.0, 37.2, 36.9, 36.5, 31.8 31.6 (2C), 31.3, 24.7, 21.4, 19.1,
16.8, 13.4; JR (solid): 2930, 2881, 1421, 1370, 1255, 1089, 955
cm.sup.-1; HRMS-ESI m/z 343.2625 ([M+H].sup.+,
C.sub.23H,.sub.50.sub.2 requires 343.2632).
##STR00172##
ent-Progesterone (57)
[0633] (78%) white solid; R.sub.f=0.41 (1:1 EA/hex);
[c].sup.23.sub.0-200.2 (c=1.00, CHCl,) [lit. -200 (c=0.25,
CHCl,)]''.sup.5 `H NIMR (400 MHz, CDCl,) .delta. 5.71 (s, 1H), 2.51
(t, 1H, J=9.0 Hz), 2.46-0.91 (m, 19H), 2.10 (s, 3H), 1.16 (s, 3H),
0.64 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.6,
199.7, 171.2, 124.1, 63.7, 56.2, 53.8, 44.1, 38.8 (2C), 35.9, 35.7,
34.1, 33.0, 32.1, 31.7, 24.6, 23.0, 21.2, 17.6, 13.5; IR (solid):
2943, 2925, 2850, 1697, 1660, 1615, 1437, 1356, 1194, 1162, 948,
871 cm.sup.-1; HRMS-ESI m/z 315.2319 ([M+H].sup.+,
C.sub.21H310.sub.2 requires 315.2319).
Enantiomeric Progesterone Derivatives
[0634] The following compounds were prepared according to the
procedures described above for the synthesis of P1-31 and related
series analogues.
##STR00173##
3.alpha.-Hydroxy-ent-progesterone (58)
[0635] (47%) white solid; R.sub.f=0.31 (99:5 DCM/MeOH, PMA stain);
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 5.29 (d, 1H, J 1.6 Hz),
4.18-4.13 (m, 1H), 2.45-0.74 (m, 21H), 2.11 (s, 3H), 1.04 (s, 3H),
0.62 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.9,
147.3, 123.8, 68.1, 63.9, 56.5, 54.5, 44.3, 39.1, 37.5, 36.1, 35.6,
33.1, 32.3, 31.7, 29.6, 24.6, 22.9, 21.2, 19.1, 13.5; IR (solid):
3400, 2932, 2848, 1701, 1662, 1355, 917, 730 cm.sup.-1, HRMS-ESI
m/z 299.2370 ([M+H-H.sub.2O].sup.+, C.sub.21H.sub.31O requires
299.2369).
##STR00174##
N-Fmoc-L-valine-3.alpha.-ent-progesterone (59)
[0636] (83%) white foam; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
7.78 (d, 2H, J 7.6 Hz), 7.61 (d, 2H, J 7.6 Hz), 7.41 (1, 2H, J 7.6
Hz), 7.33 (t, 2H, J 7.6 Hz), 5.34 (d, 1H, J=9.2 Hz), 5.31 (s, 1H),
5.24 (s, 1H), 4.40 (d, 2H, J 7.2 Hz), 4.31 (dd, 1H, J 9.2, 4.8 Hz),
4.25 (t, 1H, J 6.8 Hz), 2.53 (t, 1H, J 8.8 Hz), 2.30-0.70 (m, 20H),
2.12 (s, 3H), 1.07 (s, 3H), 0.99 (d, 3H, J 6.8 Hz), 0.93 (d, 3H, J
7.2 Hz), 0.64 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta.
209.8, 172.1, 156.5, 149.8, 144.2, 144.0, 141.5, 127.9, 127.3,
125.3, 120.2, 119.0, 72.1, 67.2, 63.7, 59.2, 56.5, 54.2, 47.4,
44.3, 39.0, 37.5, 36.0, 35.1, 33.0, 32.3, 31.6, 25.3, 24.6, 23.0,
19.2, 19.0, 17.7, 13.6; IR (solid): 3360, 2935, 1702, 1388, 1351,
1205, 734 cm.sup.-1.
##STR00175##
3.alpha.-L-Valine-ent-progesterone (60)
[0637] (87%) clear oil; .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
5.32-5.28 (m, 2H), 5.23 (d, 1H, J 0.8 Hz), 3.28 (d, 1H, J 5.2 Hz),
2.53 (t, 1H, J 9.2 Hz), 2.30-0.75 (m, 21H), 2.12 (s, 3H), 1.07 (s,
3H), 0.98 (d, 3H, J 7.2 Hz), 0.90 (d, 3H, J 6.8 Hz), 0.64 (s, 3H);
.sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 209.8, 175.7, 149.5,
119.3, 71.3, 63.9, 60.2, 56.5, 54.2, 44.3, 39.0, 37.5, 36.0, 35.1,
33.0, 32.3 (2C), 31.8, 25.3, 24.6, 23.0, 21.2, 19.6, 19.0, 17.3,
13.6; IR (solid): 2933, 2850, 1724, 1705, 1385, 1357, 1178, 977
cm.sup.-1; HRMS-ESI m/z 416.3158 ([M+H].sup.+,
C.sub.26H.sub.42NO.sub.3 requires 416.3159).
##STR00176##
3.alpha.-L-Valine-ent-progesterone-HCl (P2-13)
[0638] (58%) off-white solid; .sup.1H NMR (400 MHz, DMSO) .delta.
8.53 (bs, 3H), 5.28 (t, 1H, J 8.0 Hz), 5.22 (s, 1H), 3.82 (d, 1H, J
4.8 Hz), 3.34 (s, 1H), 2.56 (t, 1H, J 9.2 Hz), 2.25-0.74 (m, 21H),
2.05 (s, 3H), 1.02 (s, 3H), 0.99 (d, 3H, J=6.8 Hz), 0.94 (d, 3H,
J=6.8 Hz), 0.54 (s, 3H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta.
209.7, 168.5, 150.0, 118.7, 73.8, 63.9, 59.2, 56.5, 54.2, 44.2,
39.1, 37.5, 36.0, 35.2, 33.1, 32.3, 31.8, 30.3, 25.4, 24.6, 23.0,
21.2, 19.2, 18.9 (2C), 13.6; IR (solid): 2932, 2849, 2600, 1737,
1702, 1379, 1354, 1219, 1110 cm.sup.-1; HRMS-ESI m/z 416.3160
([M+H]+, C.sub.26H.sub.42NO.sub.3 requires 416.3159); Anal. Calcd
for C.sub.26H.sub.41ClNO.sub.3+1/2H.sub.2O: C, 67.73; H, 9.40; N,
3.04. Found C, 67.39; H, 9.24; N, 3.01.
Example 7
Solubility of Steroid Analogues
[0639] Several compounds were screened for solubility (FIG. 2).
Solubility testing was done in phosphate buffered saline, which is
considered to be a good model system for physiological conditions.
Compounds were added in small portions at room temperature with
stirring until a visible endpoint of saturation was reached.
Neither PROG nor ALLO showed any degree of solubility by this
method (designated at <0-05 mg/mL solubility). The valine
coupled C-3-.beta.-hydroxy PROG derivative P1-31 showed the best
solubility (3.85 mg/mL) of the different amino acid substituted
analogues within that series. Compound P1-133, the 3.alpha.-5.beta.
ALLO isomer, showed the highest solubility within that group, (0.33
mg/mL). Both the E and Z oxime derivatives P1-185 and P1-186 showed
excellent solubility, with values of 13.0 and 15.2 mg/mL
respectively.
Example 8
In Vitro Screening
[0640] Primary cortical cells were seeded in multi-well plates and
cultured for 8 days. Cells were then pre-treated with various
concentrations of different PROG analogues (0.1, 1, 5, 10, 20, 40,
and 80 .mu.M) for 24 h. Cells were next exposed to glutamate (0.5
.mu.M) for the following 24 h. Cytotoxicity was assessed by the MTT
[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]
assay, which is based on the ability of a mitochondrial
dehydrogenase enzyme from viable cells to cleave the tetrazolium
rings of the pale yellow MTT. This reaction forms dark blue
formazan crystals which are largely impermeable to cell membranes,
thus resulting in their accumulation within healthy cells.
Solubilization of the cells results in the liberation of the
crystals, which are then also solubilized. The number of surviving
cells is directly proportional to the level of the formazan product
created.
[0641] Table 2 below shows the effect of several compounds of the
invention compared with PROG and ALLO in the reduction in cortical
neuron cell death caused by glutamate toxicity. Several of the
derivatives proved to be significantly more potent than PROG or
ALLO when the compounds are compared at the 5 .mu.M concentration.
The P1-185 and P1-186 oxime prodrug compounds achieved the highest
levels of cell survival among the compounds screened. The C-20
reduced PROG derivative P1-57 and the ent-PROG derivative P2-13
also showed significant reductions in cell death.
TABLE-US-00005 TABLE 2 Reduction in cortical neuron cell death
caused by glutamate toxicity. reduction in cell death reduction in
cell best concentration death at 5 .mu.M compound (%) (%) PROG 42
(20 .mu.M) 4 ALLO 40 (80 .mu.M) -3 P1-57 30 (10 .mu.M) 23 P1-185 27
(5 .mu.M) 27 P1-186 34 (5 .mu.M) 34 P2-13 26 (5 .mu.M) 26
Example 9
Pharmacokinetic Data
[0642] Select compounds were submitted for pharmacokinetic analysis
in a non-injury whole animal rat study. The C-3-.beta.-hydroxy PROG
derivatives P1-31 and P1-33 were chosen in order to observe any
potential differences in the stability of the compounds that may be
attributable to the amino acid component. The natural ALLO
derivative P1-131 was selected based on its activity in the
cerebral edema assay and as an example of a compound containing a
saturated A ring. Finally, the oxime derived compound P1-186 was
also submitted in order to evaluate its potential to act as a
prodrug in vivo as envisioned.
[0643] The compounds were intravenously (IV) dosed at 10 mg/kg and
serum samples were taken at several time points over the course of
12 h. Serum concentration levels were determined for the analogues,
as well as for their respective parent compound. The derivatives
P1-31 and P1-33 behaved very similarly in vivo (FIG. 7). The amino
acid side chain in both compounds was cleaved almost immediately to
give rapid conversion to their mutual parent, the
C-3-.beta.-hydroxy compound 3. However, in the case of the ALLO
derivative P1-131, the valine tethered compound was stable for a
prolonged period of time and only gradually converted to its
parent, ALLO (2, FIG. 4). The data indicates a distinct difference
in the susceptibility of the amino acid side chain to hydrolysis
resulting from relatively small differences in saturation and
conformation. The more sterically congested A-ring of P1-131 could
be thought to provide some protection to active esterases relative
to the unsaturated A-ring of P1-31 type compounds, or perhaps also
important is the relative .alpha.-(P1-131) or .beta.-(P1-31) face
orientation of the side chain.
[0644] The oxime based compound P1-186 did indeed generate PROG in
vivo when dosed either IV or intraperitoneally (IP) at 10 mg/kg
(FIG. 5). The PROG levels observed via both modes of administration
reached a maximum of approximately 100 ng/mL, which compares
favorably to a previously reported maximum serum concentration of
28 ng/mL when PROG was administered IP at 4 mg/kg to male rats
(Wright et al., J. Neurotrama, 2001, 18, 901-094). Despite the
observance of PROG however, the amino acid tether of compound
P1-186 is readily cleaved to give primarily the parent oxime P2-02
(FIG. 8), which is stable in vivo for several hours. The potential
implication of this data is that it is the intermediate oxime
compound (P2-02) that may be responsible for achieving the
neuroprotective effects observed in both the in vitro and cerebral
edema assays, not progesterone, as might have been suspected.
[0645] The above embodiments are provided merely as non-limiting
examples to which the invention should not be limited.
* * * * *