U.S. patent application number 13/470377 was filed with the patent office on 2012-09-06 for method for treating central nervous system disorders.
This patent application is currently assigned to SUCAMPO AG. Invention is credited to Ryuji UENO.
Application Number | 20120225938 13/470377 |
Document ID | / |
Family ID | 36740907 |
Filed Date | 2012-09-06 |
United States Patent
Application |
20120225938 |
Kind Code |
A1 |
UENO; Ryuji |
September 6, 2012 |
METHOD FOR TREATING CENTRAL NERVOUS SYSTEM DISORDERS
Abstract
The present invention provides a method for treating a central
nervous system disorder in a mammalian subject, which comprises
administering an effective amount of a 11-deoxy-prostaglandin
compound to a subject in need thereof. The invention also provide
novel 11-deoxy-prostaglandin compound.
Inventors: |
UENO; Ryuji; (Potomac,
MD) |
Assignee: |
SUCAMPO AG
Zug
CH
|
Family ID: |
36740907 |
Appl. No.: |
13/470377 |
Filed: |
May 14, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11339495 |
Jan 26, 2006 |
8202909 |
|
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13470377 |
|
|
|
|
60647008 |
Jan 27, 2005 |
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Current U.S.
Class: |
514/530 ;
514/559; 514/573; 554/222; 562/503 |
Current CPC
Class: |
A61K 31/557 20130101;
A61P 11/00 20180101; A61P 21/04 20180101; A61P 25/06 20180101; A61P
9/10 20180101; A61P 25/28 20180101; C07C 405/00 20130101; A61P
25/14 20180101; A61P 25/00 20180101; A61P 25/02 20180101; A61P 9/00
20180101; A61P 25/16 20180101; A61P 29/00 20180101; A61P 9/02
20180101; A61P 25/36 20180101; A61P 25/08 20180101; A61P 35/00
20180101 |
Class at
Publication: |
514/530 ;
514/573; 562/503; 514/559; 554/222 |
International
Class: |
A61K 31/19 20060101
A61K031/19; A61K 31/215 20060101 A61K031/215; A61K 31/20 20060101
A61K031/20; A61P 25/00 20060101 A61P025/00; C07C 405/00 20060101
C07C405/00 |
Claims
1. A method for protecting cerebrovascular endothelial cells in a
mammalian subject, which comprises administering an effective
amount of an 11-deoxyprostaglandin compound represented by the
formula (III): ##STR00022## wherein L is hydrogen, hydroxy,
halogen, lower alkyl, hydroxy(lower)alkyl, lower alkanoyloxy or
oxo, wherein the five-membered ring may optionally have at least
one double bond; A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH,
--COOH or a functional derivative thereof; B is single bond,
--CH.sub.2--CH.sub.2--, --CH--CH--, --C.ident.C--,
--CH.sub.2--CH.sub.2--CH.sub.2--, --CH.dbd.CH--CH.sub.2--,
--CH.sub.2--CH.dbd.CH--, --C.ident.C--CH.sub.2-- or
--CH.sub.2--C.ident.C--; C.dbd.Z is ##STR00023## wherein R.sub.4
and R.sub.5 are hydrogen, hydroxy, halogen, lower alkyl, lower
alkoxy or hydroxy(lower)alkyl, wherein R.sub.4 and R.sub.5 are not
hydroxy and lower alkoxy at the same time; X.sub.1 and X.sub.2 are
hydrogen, lower alkyl, or halogen; R.sub.1 is a saturated or
unsaturated bivalent lower or medium aliphatic hydrocarbon, which
is unsubstituted or substituted with halogen, alkyl, hydroxy, oxo,
aryl or heterocyclic group, and at least one of carbon atom in the
aliphatic hydrocarbon is optionally substituted by oxygen, nitrogen
or sulfur; and R.sub.2 is a single bond or lower alkylene; and
R.sub.3 is lower alkyl, lower alkoxy, lower alkanoyloxy,
cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group, and at least one of
carbon atom in the aliphatic hydrocarbon is optionally substituted
by oxygen, nitrogen or sulfur, provided that one of X.sub.1 and
X.sub.2 is substituted by halogen and/or C.dbd.Z is C.dbd.O; to a
subject in need thereof.
2. The method described in claim 1, wherein the cerebrovascular
endothelial cell is that of a Blood-brain barrier.
3. A compound represented by the formula (IV): ##STR00024## wherein
L is hydrogen, hydroxy, halogen, lower alkyl, hydroxy(lower)alkyl,
lower alkanoyloxy or oxo, wherein the five-membered ring may
optionally have at least one double bond; A is --CH.sub.3,
--CH.sub.2OH, --COCH.sub.2OH, --COOH or a functional derivative
thereof; B is single bond, --CH.sub.2--CH.sub.2--, --CH.dbd.CH--,
--C.ident.C--, --CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.dbd.CH--,
--C.ident.C--CH.sub.2-- or --CH.sub.2--C.ident.C--; C.dbd.Z is
##STR00025## wherein R.sub.4 and R.sub.5 are hydrogen, hydroxy,
halogen, lower alkyl, lower alkoxy or hydroxy(lower)alkyl, wherein
R.sub.4 and R.sub.5 are not hydroxy and lower alkoxy at the same
time; X.sub.1' and X.sub.2' are same or different halogen atoms;
R.sub.1 is a saturated or unsaturated bivalent lower or medium
aliphatic hydrocarbon, which is unsubstituted or substituted with
halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and at
least one of carbon atom in the aliphatic hydrocarbon is optionally
substituted by oxygen, nitrogen or sulfur; and R.sub.2 is a single
bond or lower alkylene; and R.sub.3 is lower alkyl, lower alkoxy,
lower alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl,
aryloxy, heterocyclic group or heterocyclic-oxy group, and at least
one of carbon atom in the aliphatic hydrocarbon is optionally
substituted by oxygen, nitrogen or sulfur; provided that the
compound is not
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1.
4. The compound as described in claim 3 selected from the group
consisting of: 11-deoxy-13,14-dihydro-16,16-difluoro-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 isopropyl
ester,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE.sub.1 isopropyl ester,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1
isopropyl ester,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 methyl
ester,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.1
isopropyl ester and
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGF.sub.1.alpha.
isopropyl ester.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is a divisional of application Ser. No. 11/339,495
filed Jan. 26, 2006, which claims the benefit of U.S. Provisional
Application No. 60/647,008 filed Jan. 27, 2005, the disclosures of
which are all hereby incorporated by reference.
TECHNICAL FIELD
[0002] The present invention relates to a method for treating a
central nervous system disorder in a mammalian subject. The
invention also relates to a novel prostaglandin compound.
BACKGROUND ART
[0003] Intercellular junctions mediate adhesion and communication
between adjoining endothelial and epithelial cells. In the
endothelium, junctional complexes comprise tight junctions,
adherens junctions, and gap junctions. The expression and
organization of these complexes depend on the type of vessels and
the permeability requirements of perfused organs. Gap junctions are
communication structures, which allow the passage of small
molecular weight solutes between neighboring cells. Tight junctions
serve the major functional purpose of providing a "barrier" and a
"fence" within the membrane, by regulating paracellular
permeability and maintaining cell polarity. Adherens junctions play
an important role in contact inhibition of endothelial cell growth,
paracellular permeability to circulating leukocytes and solutes. In
addition, they are required for a correct organization of new
vessels in angiogenesis (Physiol. Rev. 84(3), 869-901, 2004).
[0004] The mechanism by which epithelial and endothelial cells
interact to form polarized tissue is of fundamental importance to
multicellular organisms. Dysregulation of these barriers occurs in
a variety of diseases, destroying the normal cellular environments
and leading to organ failure.
[0005] Cerebral microvascular endothelial cells that form the
blood-brain barrier (BBB) have tight junctions that are critical
for maintaining brain homeostasis and low permeability.
[0006] The blood-brain barrier (BBB) is a specialized structure in
the central nervous system (CNS), which participates in maintenance
of a state of cerebrospinal fluid homeostasis by controlling the
access of nutrients and toxic substances to the central nervous
system (CNS).
[0007] The base membrane underlying the vasculature plays a
critical role in maintaining the integrity of the BBB by providing
structural support to the endothelial cell wall (Trends Neurosci.
1990; 13(5): 174-178). The BBB serves to protect the central
nervous system (CNS) from invasive agents, such as inflammatory
cells and bacteria, as well as from chemical agents.
[0008] A wide range of central nervous system (CNS) disorders
associated with disruption of the BBB are known. Examples of the
disorders include multiple sclerosis, experimental allergic
encephalomyelitis, bacterial meningitis, ischemia, brain edema,
Alzheimer's disease, acquired immune deficiency syndrome dementia
complex (Helga E. DE Vries et al, Pharmacological Reviews, 49(2):
143-155, 1997), brain tumors (Davies D. C. et al., J. Anat., 200
(6): 639-46, 2002), traumatic brain injury (Hartl R et. al., Acta
Neurochir Suppl. 70: 240-242, 1997).
[0009] It has also been reported that, after focal stroke, there is
a breakdown of the BBB with an associated increase in vascular
permeability. Damage to the BBB often results in hemorrhage and
edema, resulting in neuronal cell death (Biomedicine. 1974;
21:36-39, Stroke, 1998; 29(5): 1020-1030, Stroke, 2003;
34(3):806-812, J. Neurotrauma. 1995; 12:833-842). Brain injury
after focal stroke is primarily a result of the decrease in blood
flow and of energy depletion due to occlusion of a cerebral blood
vessel. The neuronal tissue becomes infracted as a result of these
events, with contributions from excitotoxicity, enzyme activation,
edema, and inflammation (Trends Pharmacol Sci. 1996; 17:227-233,
Crit. Care Med. 1988; 16:954-963).
[0010] Furthermore, systemic-derived inflammation has recently been
shown to cause BBB tight junctional disruption and increased
paracellular permeability. The BBB is capable of rapid modulation
in response to physiological stimuli at the cytoskeletal level,
which enables it to protect the brain parenchyma and maintain a
homeostatic environment.
[0011] Research has shown that destruction of the BBB is associated
with diseases of the CNS. However, there is little research on how
the BBB might be protected.
[0012] Prostaglandins (hereinafter, referred to as PG(s)) are
members of class of organic carboxylic acids, which are contained
in tissues or organs of human or other mammals, and exhibit a wide
range of physiological activity. PGs found in nature (primary PGs)
generally have a prostanoic acid skeleton as shown in the formula
(A):
##STR00001##
[0013] On the other hand, some of synthetic analogues of primary
PGs have modified skeletons. The primary PGs are classified into
PGAs, PGBs, PGCs, PGDs, PGEs, PGFs, PGGs, PGHs, PGIs and PGJs
according to the structure of the five-membered ring moiety, and
further classified into the following three types by the number and
position of the unsaturated bond at the carbon chain moiety:
[0014] Subscript 1: 13,14-unsaturated-15-OH
[0015] Subscript 2: 5,6- and 13,14-diunsaturated-15-OH
[0016] Subscript 3: 5,6-, 13,14-, and
17,18-triunsaturated-15-OH.
[0017] Further, the PGFs are classified, according to the
configuration of the hydroxyl group at the 9-position, into a type
(the hydroxyl group is of an .alpha.-configuration) and .beta.-type
(the hydroxyl group is of a .beta.-configuration).
[0018] PGE.sub.1 and PGE.sub.2 and PGE.sub.3 are known to have
vasodilation, hypotension, gastric secretion decreasing, intestinal
tract movement enhancement, uterine contraction, diuretic,
bronchodilation and anti ulcer activities. PGF.sub.1.alpha.,
PGF.sub.2.alpha. and PGF.sub.3.alpha. have been known to have
hypertension, vasoconstriction, intestinal tract movement
enhancement, uterine contraction, lutein body atrophy and
bronchoconstriction activities.
[0019] Some 15-keto (i.e., having oxo at the 15-position instead of
hydroxy)-PGs and 13,14-dihydro (i.e., having single bond between
the 13 and 14-position)-15-keto-PGs are known as the substances
naturally produced by the action of enzymes during the metabolism
of primary PGs.
[0020] U.S. Pat. No. 5,290,811 to Ueno et al. describes that some
15-keto-PG compounds are useful for improvement of encephalic
function. U.S. Pat. No. 5,290,811 indicates that when the bond
between 13- and 14-positions is saturated, a keto-hemiacetal
equilibrium may sometimes be formed by the formation of a
hemiacetal between the hydroxy group at 11-position and the keto
group at 15-position.
[0021] U.S. Pat. No. 5,317,032 to Ueno et al. describes
prostaglandin compound cathartics, including the existence of
bicyclic tautomers and U.S. Pat. No. 6,414,016 to Ueno describes
the bicyclic tautomers as having pronounced activity as
anti-constipation agents. The bicyclic tautomers, substituted by
one or more halogen atoms can be employed in small doses for
relieving constipation. At the C-16 position, especially, fluorine
atoms can be employed in small doses for relieving
constipation.
SUMMARY OF THE INVENTION
[0022] The present inventor conducted an intensive study and found
that 11-deoxy-prostaglandin compounds possessed significant effects
on the central nervous system disorders, which resulted in the
completion of the present invention.
[0023] Namely, the present invention relates to a method for
treating a central nervous system disorder in a mammalian subject,
which comprises administering an effective amount of a
11-deoxy-prostaglandin compound to a subject in need thereof.
[0024] The present invention further relates to a composition for
treating a central nervous system disorder in a mammalian subject,
which comprises an effective amount of a 11-deoxy-prostaglandin
compound.
[0025] Furthermore, the present invention relates to a use of
11-deoxy-prostaglandin compound for manufacturing a composition for
treating a central nervous system disorder in a mammalian subject,
which comprises an effective amount of a 11-deoxy-prostaglandin
compound.
[0026] Another embodiment of the present invention relates to a
method for protecting cerebrovascular endothelial cells in a
mammalian subject, which comprises administering an effective
amount of a 11-deoxy-prostaglandin compound to a subject in need
thereof.
[0027] In another aspect of the present invention, a novel compound
represented by the formula (IV):
##STR00002##
[0028] wherein L is hydrogen, hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, lower alkanoyloxy or oxo; wherein the
five-membered ring may optionally have at least one double
bond;
[0029] A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH, --COOH or a
functional derivative thereof;
[0030] B is single bond, --CH.sub.2--CH.sub.2--, --CH.dbd.CH--,
--C.ident.C--, --CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.dbd.CH--,
--C.ident.C--CH.sub.2-- or --CH.sub.2--C.dbd.C--;
[0031] Z is
##STR00003##
[0032] wherein R.sub.4 and R.sub.5 are hydrogen, hydroxy, halogen,
lower alkyl, lower alkoxy or hydroxy(lower)alkyl, wherein R.sub.4
and R.sub.5 are not hydroxy and lower alkoxy at the same time;
[0033] X.sub.1' and X.sub.2' are same or different halogen
atoms;
[0034] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon, which is unsubstituted or substituted
with halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and
at least one of carbon atom in the aliphatic hydrocarbon is
optionally substituted by oxygen, nitrogen or sulfur;
[0035] R.sub.2 is a single bond or lower alkylene; and
[0036] R.sub.3 is lower alkyl, lower alkoxy, lower alkanoyloxy,
cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group, and at least one of
carbon atom in the aliphatic hydrocarbon is optionally substituted
by oxygen, nitrogen or sulfur;
[0037] provided that the formula (IV) is not
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 is
provided.
BRIEF DESCRIPTION OF THE DRAWINGS
[0038] FIG. 1 is a graph showing the effect of Compound A on
Recovery of Transendothelial Electrical Resistance (TEER). Human
vascular endothelial cell cultures were brought to confluence, as
measured by transendothelial electrical resistance (TEER). The cell
cultures were then deprived of oxygen for 30 minutes by incubation
in a nitrogen atmosphere. The cells were then either treated with
0.1% DMSO or with 5 nM Compound A with 0.1% DMSO. Statistical
significance is indicated at all data points after drug treatment.
N=10 cells.
[0039] FIG. 2 is a graph showing the effect of Compound A on
Recovery of ATP Level. Human microvascular endothelial cells
(adult) (HMVEC-AD) were grown to confluence. The cells were then
treated for 30 minutes with a nitrogen atmosphere and returned to
normal oxygen. ATP levels were monitored at the indicated time
points using a luciferin-luciferase assay system (ATPlite, Perkin
Elmer). ATP levels are given as relative luminescence. N=6 cells at
each time point.
[0040] FIG. 3 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (6) obtained in Synthesis Example 2 below.
[0041] FIG. 4 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (6) obtained in Synthesis Example 2 below.
[0042] FIG. 5 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (9) obtained in Synthesis Example 3 below.
[0043] FIG. 6 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (9) obtained in Synthesis Example 3 below.
[0044] FIG. 7 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (12) obtained in Synthesis Example 4 below.
[0045] FIG. 8 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (12) obtained in Synthesis Example 4 below.
[0046] FIG. 9 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (15) obtained in Synthesis Example 5 below.
[0047] FIG. 10 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (15) obtained in Synthesis Example 5 below.
[0048] FIG. 11 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (18) obtained in Synthesis Example 6 below.
[0049] FIG. 12 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (18) obtained in Synthesis Example 6 below.
[0050] FIG. 13 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (21) obtained in Synthesis Example 7 below.
[0051] FIG. 14 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (21) obtained in Synthesis Example 7 below.
[0052] FIG. 15 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (23) obtained in Synthesis Example 8 below.
[0053] FIG. 16 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (23) obtained in Synthesis Example 8 below.
[0054] FIG. 17 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (25) obtained in Synthesis Example 9 below.
[0055] FIG. 18 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (25) obtained in Synthesis Example 9 below.
[0056] FIG. 19 is a .sup.1H-NMR (200 MHz, CDCl.sub.3) chart of the
compound (34) obtained in Synthesis Example 10 below.
[0057] FIG. 20 is a .sup.13C-NMR (50 MHz, CDCl.sub.3) chart of the
compound (34) obtained in Synthesis Example 10 below.
DETAILED DESCRIPTION OF THE INVENTION
[0058] In the present invention, the "11-deoxy-prostaglandin
compound" (hereinafter, referred to as "11-deoxy-PG compound") may
include any derivatives or analogs (including substituted
derivatives) of a compound having no substituent at 11-position of
the prostanoic acid skeleton, irrespective of the configuration of
the five-membered ring, the number of double bonds, presence or
absence of a substituent, or any other modification in the .alpha.
or .omega. chain.
[0059] The formula (A) shows a basic skeleton of the C-20 carbon
atoms, but the present invention is not limited to those having the
same number of carbon atoms. In the formula (A), the numbering of
the carbon atoms which constitute the basic skeleton of the PG
compounds starts at the carboxylic acid (numbered 1), and carbon
atoms in the .alpha.-chain are numbered 2 to 7 towards the
five-membered ring, those in the ring are 8 to 12, and those in the
.omega.-chain are 13 to 20. When the number of carbon atoms is
decreased in the .alpha.-chain, the number is deleted in the order
starting from position 2; and when the number of carbon atoms is
increased in the .alpha.-chain, compounds are named as substitution
compounds having respective substituents at position 2 in place of
the carboxy group (C-1). Similarly, when the number of carbon atoms
is decreased in the w-chain, the number is deleted in the order
starting from position 20; and when the number of carbon atoms is
increased in the .omega.-chain, the carbon atoms beyond position 20
are named as substituents. Stereochemistry of the compounds is the
same as that of the above formula (A) unless otherwise
specified.
[0060] As stated above, the nomenclature of the 11-deoxy-PG
compounds is based on the prostanoic acid skeleton. However, in
case the compound has a similar partial structure as a
prostaglandin, the abbreviation of "PG" may be used. Thus, a
11-deoxy-PG compound of which .alpha.-chain is extended by two
carbon atoms, that is, having 9 carbon atoms in the .alpha.-chain
is named as 2-decarboxy-2-(2-carboxyethyl)-1'-deoxy-PG compound.
Similarly, 11-deoxy-PG compound having 11 carbon atoms in the
.alpha.-chain is named as
2-decarboxy-2-(4-carboxybutyl)-11-deoxy-PG compound. Further,
11-deoxy-PG compound of which .alpha.-chain is extended by two
carbon atoms, that is, having 10 carbon atoms in the .alpha.-chain
is named as 11-deoxy-20-ethyl-PG compound. These compounds,
however, may also be named according to the IUPAC
nomenclatures.
[0061] Examples of the analogs (including substituted derivatives)
or derivatives include a 11-deoxy-PG compound of which carboxy
group at the end of .alpha.-chain is esterified; a compound of
which .alpha.-chain is extended; physiologically acceptable salt
thereof; a compound having a double bond at 2-3 position or a
triple bond at position 5-6, a compound having substituent(s) at
position 3, 5, 6, 16, 17, 18, 19 and/or 20; and a compound having
lower alkyl or a hydroxy (lower) alkyl group at position 9 in place
of the hydroxy group.
[0062] According to the present invention, preferred substituents
at position 3, 17, 18 and/or 19 include alkyl having 1-4 carbon
atoms, especially methyl and ethyl. Preferred substituents at
position 16 include lower alkyl such as methyl and ethyl, hydroxy,
halogen atoms such as chlorine and fluorine, and aryloxy such as
trifluoromethylphenoxy. Preferred substituents at position 17
include lower alkyl such as methyl and ethyl, hydroxy, halogen
atoms such as chlorine and fluorine, aryloxy such as
trifluoromethylphenoxy. Preferred substituents at position 20
include saturated or unsaturated lower alkyl such as C1-4 alkyl,
lower alkoxy such as C1-4 alkoxy, and lower alkoxy alkyl such as
C1-4 alkoxy-C1-4 alkyl. Preferred substuents at position 5 include
halogen atoms such as chlorine and fluorine. Preferred substituents
at position 6 include an oxo group forming a carbonyl group.
Stereochemistry of PGs having hydroxy, lower alkyl or
hydroxy(lower)alkyl substituent at position 9 may be .alpha.,
.beta. or a mixture thereof.
[0063] Further, the above analogs or derivatives may be compounds
having an alkoxy, cycloalkyl, cycloalkyloxy, phenoxy or phenyl
group at the end of the .omega.-chain where the chain is shorter
than the primary PGs.
[0064] The nomenclature of the 11-deoxy-PG compounds used herein is
based on the numbering system of the prostanoic acid represented in
the above formula (A).
[0065] A preferred compound used in the present invention is
represented by the formula (I):
##STR00004##
[0066] wherein L and N are hydrogen, hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, lower alkanoyloxy or oxo, wherein the
five-membered ring may optionally have at least one double
bond;
[0067] A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH, --COOH or a
functional derivative thereof;
[0068] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon, which is unsubstituted or substituted
with halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and
at least one of carbon atom in the aliphatic hydrocarbon is
optionally substituted by oxygen, nitrogen or sulfur; and
[0069] R.sub.0 is a saturated or unsaturated lower or medium
aliphatic hydrocarbon residue, which is unsubstituted or
substituted with halogen, oxo, hydroxy, lower alkyl, lower alkoxy,
lower alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl,
aryloxy, heterocyclic group or hetrocyclic-oxy group; lower alkoxy;
lower alkanoyloxy; cyclo(lower)alkyl; cyclo(lower)alkyloxy; aryl;
aryloxy; heterocyclic group; heterocyclic-oxy group, and at least
one of carbon atom in the aliphatic hydrocarbon is optionally
substituted by oxygen, nitrogen or sulfur.
[0070] A more preferred compound used in the present invention is
represented by the formula (II):
##STR00005##
[0071] wherein L and N are hydrogen, hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, lower alkanoyloxy or oxo, wherein the
five-membered ring may optionally have at least one double
bond;
[0072] A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH, --COOH or a
functional derivative thereof;
[0073] B is single bond, --CH.sub.2--CH.sub.2--, --CH.dbd.CH--,
--CH.sub.2--CH.sub.2--CH.sub.2--, --CH.dbd.CH--CH.sub.2--,
--CH.sub.2--CH.dbd.CH--, --C.ident.C--CH.sub.2-- or
--CH.sub.2--C.ident.C--;
[0074] Z is
##STR00006##
[0075] wherein R.sub.4 and R.sub.5 are hydrogen, hydroxy, halogen,
lower alkyl, lower alkoxy or hydroxy(lower)alkyl, wherein R.sub.4
and R.sub.5 are not hydroxy and lower alkoxy at the same time;
[0076] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon, which is unsubstituted or substituted
with halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and
at least one of carbon atom in the aliphatic hydrocarbon is
optionally substituted by oxygen, nitrogen or sulfur; and
[0077] Ra is a saturated or unsaturated lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen, oxo, hydroxy, lower alkyl, lower alkoxy, lower
alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl,
aryloxy, heterocyclic group or hetrocyclic-oxy group; lower alkoxy;
lower alkanoyloxy; cyclo(lower)alkyl; cyclo(lower)alkyloxy; aryl;
aryloxy; heterocyclic group; heterocyclic-oxy group, and at least
one of carbon atom in the aliphatic hydrocarbon is optionally
substituted by oxygen, nitrogen or sulfur.
[0078] A group of particularly preferable compounds among the
above-described compounds is represented by the formula (III):
##STR00007##
[0079] wherein L is hydrogen, hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, lower alkanoyloxy or oxo, wherein, and the
five-membered ring may optionally have at least one double
bond;
[0080] A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH, --COOH or a
functional derivative thereof;
[0081] B is single bond, --CH.sub.2--CH.sub.2--, --CH.dbd.CH--,
--C.dbd.C--, --CH.sub.2--CH.sub.2--CH.sub.2--, --CH'CH--CH.sub.2--,
--CH.sub.2--CH.dbd.CH--, --C.ident.C--CH.sub.2-- or
--CH.sub.2--C.ident.C--;
[0082] Z is
##STR00008##
[0083] wherein R.sub.4 and R.sub.5 are hydrogen, hydroxy, halogen,
lower alkyl, lower alkoxy or hydroxy(lower)alkyl, wherein R.sub.4
and R.sub.5 are not hydroxy and lower alkoxy at the same time;
[0084] X.sub.1 and X.sub.2 are hydrogen, lower alkyl, or
halogen;
[0085] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon, which is unsubstituted or substituted
with halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and
at least one of carbon atom in the aliphatic hydrocarbon is
optionally substituted by oxygen, nitrogen or sulfur; and
[0086] R.sub.2 is a single bond or lower alkylene; and
[0087] R.sub.3 is lower alkyl, lower alkoxy, lower alkanoyloxy,
cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group, and at least one of
carbon atom in the aliphatic hydrocarbon is optionally substituted
by oxygen, nitrogen or sulfur.
[0088] The present invention further relates to a compound
represented by the formula (IV):
##STR00009##
[0089] wherein L is hydrogen, hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, lower alkanoyloxy or oxo, wherein the
five-membered ring may optionally have at least one double
bond;
[0090] A is --CH.sub.3, --CH.sub.2OH, --COCH.sub.2OH, --COOH or a
functional derivative thereof;
[0091] B is single bond, --CH.sub.2--CH.sub.2--, --CH.dbd.CH--,
--CH.sub.2--CH.sub.2--CH.sub.2--, --CH.dbd.CH--CH.sub.2--,
--CH.sub.2--CH.dbd.CH--, --C.ident.C--CH.sub.2-- or
--CH.sub.2--C.ident.C--;
[0092] Z is
##STR00010##
[0093] wherein R.sub.4 and R.sub.5 are hydrogen, hydroxy, halogen,
lower alkyl, lower alkoxy or hydroxy(lower)alkyl, wherein R.sub.4
and R.sub.5 are not hydroxy and lower alkoxy at the same time;
[0094] X.sub.1' and X.sub.2' are same or different halogen
atoms;
[0095] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon, which is unsubstituted or substituted
with halogen, alkyl, hydroxy, oxo, aryl or heterocyclic group, and
at least one of carbon atom in the aliphatic hydrocarbon is
optionally substituted by oxygen, nitrogen or sulfur;
[0096] R.sub.2 is a single bond or lower alkylene; and
[0097] R.sub.3 is lower alkyl, lower alkoxy, lower alkanoyloxy,
cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group, and at least one of
carbon atom in the aliphatic hydrocarbon is optionally substituted
by oxygen, nitrogen or sulfur;
[0098] provided that the formula (IV) is not
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1, and a
method for producing the same.
[0099] In the above formula, the term "unsaturated" in the
definitions for R.sub.1 and Ra is intended to include at least one
or more double bonds and/or triple bonds that are isolatedly,
separately or serially present between carbon atoms of the main
and/or side chains. According to the usual nomenclature, an
unsaturated bond between two serial positions is represented by
denoting the lower number of the two positions, and an unsaturated
bond between two distal positions is represented by denoting both
of the positions.
[0100] The term "lower or medium aliphatic hydrocarbon" refers to a
straight or branched chain hydrocarbon group having 1 to 14 carbon
atoms (for a side chain, 1 to 3 carbon atoms are preferable) and
preferably 1 to 10, especially 6 to 10 carbon atoms for R.sub.1 and
1 to 10, especially 1 to 8 carbon atoms for R.sub.a.
[0101] The term "halogen" covers fluorine, chlorine, bromine and
iodine.
[0102] The term "lower" throughout the specification is intended to
include a group having 1 to 6 carbon atoms unless otherwise
specified.
[0103] The term "lower alkyl" refers to a straight or branched
chain saturated hydrocarbon group containing 1 to 6 carbon atoms
and includes, for example, methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, t-butyl, pentyl and hexyl.
[0104] The term "lower alkoxy" refers to a group of lower
alkyl-O--, wherein lower alkyl is as defined above.
[0105] The term "hydroxy(lower)alkyl" refers to a lower alkyl as
defined above which is substituted with at least one hydroxy group
such as hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl and
1-methyl-1-hydroxyethyl.
[0106] The term "lower alkanoyloxy" refers to a group represented
by the formula RCO--O--, wherein RCO-- is an acyl group formed by
oxidation of a lower alkyl group as defined above, such as
acetyl.
[0107] The term "cyclo(lower)alkyl" refers to a cyclic group formed
by cyclization of a lower alkyl group as defined above but contains
three or more carbon atoms, and includes, for example, cyclopropyl,
cyclobutyl, cyclopentyl and cyclohexyl.
[0108] The term "cyclo(lower)alkyloxy" refers to the group of
cyclo(lower)alkyl-O--, wherein cyclo(lower)alkyl is as defined
above.
[0109] The term "aryl" may include unsubstituted or substituted
aromatic hydrocarbon rings (preferably monocyclic groups), for
example, phenyl, tolyl, xylyl. Examples of the substituents are
halogen atom and halo(lower)alkyl, wherein halogen atom and lower
alkyl are as defined above.
[0110] The term "aryloxy" refers to a group represented by the
formula ArO-, wherein Ar is aryl as defined above. The term
"heterocyclic group" may include mono- to tri-cyclic, preferably
monocyclic heterocyclic group which is 5 to 14, preferably 5 to 10
membered ring having optionally substituted carbon atom and 1 to 4,
preferably 1 to 3 of 1 or 2 type of hetero atoms selected from
nitrogen atom, oxygen atom and sulfur atom. Examples of the
heterocyclic group include furyl, thienyl, pyrrolyl, oxazolyl,
isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl,
furazanyl, pyranyl, pyridyl, pyridazinyl, pyrimidyl, pyrazinyl,
2-pyrrolinyl, pyrrolidinyl, 2-imidazolinyl, imidazolidinyl,
2-pyrazolinyl, pyrazolidinyl, piperidino, piperazinyl, morpholino,
indolyl, benzothienyl, quinolyl, isoquinolyl, purinyl,
quinazolinyl, carbazolyl, acridinyl, phenanthridinyl,
benzimidazolyl, benzimidazolinyl, benzothiazolyl, phenothiazinyl.
Examples of the substituent in this case include halogen, and
halogen substituted lower alkyl group, wherein halogen atom and
lower alkyl group are as described above.
[0111] The term "heterocyclic-oxy group" means a group represented
by the formula HcO--, wherein Hc is a heterocyclic group as
described above.
[0112] The term "functional derivative" of A includes salts
(preferably pharmaceutically acceptable salts), ethers, esters and
amides.
[0113] Suitable "pharmaceutically acceptable salts" include
conventionally used non-toxic salts, for example a salt with an
inorganic base such as an alkali metal salt (such as sodium salt
and potassium salt), an alkaline earth metal salt (such as calcium
salt and magnesium salt), an ammonium salt; or a salt with an
organic base, for example, an amine salt (such as methylamine salt,
dimethylamine salt, cyclohexylamine salt, benzylamine salt,
piperidine salt, ethylenediamine salt, ethanolamine salt,
diethanolamine salt, triethanolamine salt,
tris(hydroxymethylamino)ethane salt, monomethyl-monoethanolamine
salt, procaine salt and caffeine salt), a basic amino acid salt
(such as arginine salt and lysine salt), tetraalkyl ammonium salt
and the like. These salts may be prepared by a conventional
process, for example from the corresponding acid and base or by
salt interchange.
[0114] Examples of the ethers include alkyl ethers, for example,
lower alkyl ethers such as methyl ether, ethyl ether, propyl ether,
isopropyl ether, butyl ether, isobutyl ether, t-butyl ether, pentyl
ether and 1-cyclopropyl ethyl ether; and medium or higher alkyl
ethers such as octyl ether, diethylhexyl ether, lauryl ether and
cetyl ether; unsaturated ethers such as oleyl ether and linolenyl
ether; lower alkenyl ethers such as vinyl ether, allyl ether; lower
alkynyl ethers such as ethynyl ether and propynyl ether;
hydroxy(lower)alkyl ethers such as hydroxyethyl ether and
hydroxyisopropyl ether; lower alkoxy (lower)alkyl ethers such as
methoxymethyl ether and 1-methoxyethyl ether; optionally
substituted aryl ethers such as phenyl ether, tosyl ether,
t-butylphenyl ether, salicyl ether, 3,4-di-methoxyphenyl ether and
benzamidophenyl ether; and aryl(lower)alkyl ethers such as benzyl
ether, trityl ether and benzhydryl ether.
[0115] Examples of the esters include aliphatic esters, for
example, lower alkyl esters such as methyl ester, ethyl ester,
propyl ester, isopropyl ester, butyl ester, isobutyl ester, t-butyl
ester, pentyl ester and 1-cyclopropylethyl ester; lower alkenyl
esters such as vinyl ester and allyl ester; lower alkynyl esters
such as ethynyl ester and propynyl ester; hydroxy(lower)alkyl ester
such as hydroxyethyl ester; lower alkoxy (lower) alkyl esters such
as methoxymethyl ester and 1-methoxyethyl ester; and optionally
substituted aryl esters such as, for example, phenyl ester, tolyl
ester, t-butylphenyl ester, salicyl ester, 3,4-di-methoxyphenyl
ester and benzamidophenyl ester; and aryl(lower)alkyl ester such as
benzyl ester, trityl ester and benzhydryl ester.
[0116] The amide of A mean a group represented by the formula
--CONR'R'', wherein each of R' and R'' is hydrogen atom, lower
alkyl, aryl, alkyl- or aryl-sulfonyl, lower alkenyl and lower
alkynyl, and include for example lower alkyl amides such as
methylamide, ethylamide, dimethylamide and diethylamide; arylamides
such as anilide and toluidide; and alkyl- or aryl-sulfonylamides
such as methylsulfonylamide, ethylsulfonyl-amide and
tolylsulfonylamide.
[0117] Preferred examples of L include hydroxy or oxo which has a
5-membered ring structure of, so called, especially PGF or PGE
type.
[0118] Preferred example A is --COON, its pharmaceutically
acceptable salt, ester or amide thereof.
[0119] Preferred example B is --CH.sub.2--CH.sub.2--, which provide
the structure of so-called, 13,14-dihydro type.
[0120] Preferred example of X.sub.1 and X.sub.2 is hydrogen, or
that at least one of them is halogen, more preferably, both of them
are halogen, especially, fluorine that provides a structure of, so
called 16,16-difluoro type.
[0121] Preferred X.sub.1' and X.sub.2' are difluoro atoms.
[0122] Preferred R.sub.1 is a hydrocarbon containing 1-10 carbon
atoms, preferably, 6-10 carbon atoms. Further, at least one of
carbon atom in the aliphatic hydrocarbon is optionally substituted
by oxygen, nitrogen or sulfur.
[0123] Examples of R.sub.1 include, for example, the following
groups: [0124]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
[0125] --CH.sub.2--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--,
[0126] --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.dbd.CH--,
[0127] --CH.sub.2--C.ident.C--CH.sub.2--CH.sub.2--CH.sub.2--,
[0128]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH(CH.sub.3)--CH.sub.2--,
[0129] --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--O--CH.sub.2--
[0130] --CH.sub.2--CH.dbd.CH--CH.sub.2--O--CH.sub.2--, [0131]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2
[0132]
--CH.sub.2--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--
[0133]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.dbd.CH--
[0134]
--CH.sub.2--C.ident.C--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
[0135]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH(CH.sub.3)---
CH.sub.2--, [0136]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--C-
H.sub.2 [0137]
--CH.sub.2--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2---
, [0138]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH--
-CH--, [0139]
--CH.sub.2--C.ident.C--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2---
, [0140]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH(-
CH.sub.3)--CH.sub.2--
[0141] Preferred Ra is a hydrocarbon containing 1-10 carbon atoms,
more preferably, 1-8 carbon atoms. Ra may have one or two side
chains having one carbon atom.
[0142] Preferred R.sub.2 is single bond, and preferred R.sub.3 is
lower alkyl. R.sub.3 may have one or two side chains having one
carbon atom.
[0143] The configuration of the ring and the .alpha.- and/or
.omega. chains in the above formula (I), (II), (III) and (IV) may
be the same as or different from that of the primary PGs. However,
the present invention also includes a mixture of a compound having
a primary type configuration and a compound of a non-primary type
configuration.
[0144] The typical example of the present compound is a
11-deoxy-13,14-dihydro-16,16-difluoro-PGE or PGF compound,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE or PGF compound,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE or PGF compound, or
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl or
ethyl-PGE or PGF compound and its derivative or analogue.
[0145] The preferred example of the present compound is
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1,
11-deoxy-13,14-dihydro-16,16-difluoro-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 isopropyl
ester,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE.sub.1 isopropyl ester,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE.sub.I,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1
isopropyl ester,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.I,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 methyl
ester,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.1
isopropyl ester or
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGF.sub.1.alpha.
isopropyl ester.
[0146] In the present invention, any of isomers such as the
individual tautomeric isomers, the mixture thereof, or optical
isomers, the mixture thereof, a racemic mixture, and other steric
isomers may be used in the same purpose.
[0147] Some of the compounds used in the present invention may be
prepared by the method disclosed in U.S. Pat. Nos. 5,073,569,
5,166,174, 5,221,763, 5,212,324, 5,739,161 and 6,242,485 (these
cited references are herein incorporated by reference).
[0148] According to the present invention, a mammalian subject may
be treated by the instant invention by administering the compound
used in the present invention. The subject may be any mammalian
subject including a human. The compound may be applied systemically
or topically. Usually, the compound may be administered by oral
administration, intravenous injection (including infusion),
subcutaneous injection, intra rectal administration, intra vaginal
administration, transdermal administration and the like.
[0149] The dose may vary depending on the strain of the animal,
age, body weight, symptom to be treated, desired therapeutic
effect, administration route, term of treatment and the like. A
satisfactory effect can be obtained by systemic administration 1-4
times per day or continuous administration at the amount of
0.00001-500 mg/kg per day, more preferably 0.0001-100 mg/kg.
[0150] The compound may preferably be formulated in a
pharmaceutical composition suitable for administration in a
conventional manner. The composition may be those suitable for oral
administration, injection or perfusion as well as it may be an
external preparation, suppository or pessary.
[0151] The composition of the present invention may further contain
physiologically acceptable additives. Said additives may include
the ingredients used with the present compounds such as excipient,
diluent, filler, resolvent, lubricant, adjuvant, binder,
disintegrator, coating agent, cupsulating agent, ointment base,
suppository base, aerozoling agent, emulsifier, dispersing agent,
suspending agent, thickener, tonicity agent, buffering agent,
soothing agent, preservative, antioxidant, corrigent, flavor,
colorant, a functional material such as cyclodextrin, and
biodegradable polymer, stabilizer. The additives are well known to
the art and may be selected from those described in general
reference books of pharmaceutics.
[0152] The amount of the above-defined compound in the composition
of the invention may vary depending on the formulation of the
composition, and may generally be 0.000001-10.0%, more
preferably0.00001-5.0%, most preferably 0.0001-1%.
[0153] Examples of solid compositions for oral administration
include tablets, troches, sublingual tablets, capsules, pills,
powders, granules and the like. The solid composition may be
prepared by mixing one or more active ingredients with at least one
inactive diluent. The composition may further contain additives
other than the inactive diluents, for example, a lubricant, a
disintegrator and a stabilizer. Tablets and pills may be coated
with an enteric or gastroenteric film, if necessary.
[0154] They may be covered with two or more layers. They may also
be adsorbed to a sustained release material, or microcapsulated.
Additionally, the compositions may be capsulated by means of an
easily degradable material such gelatin. They may be further
dissolved in an appropriate solvent such as fatty acid or its mono,
di or triglyceride to be a soft capsule. Sublingual tablet may be
used in need of fast-acting property.
[0155] Examples of liquid compositions for oral administration
include emulsions, solutions, suspensions, syrups and elixirs and
the like. Said composition may further contain a conventionally
used inactive diluentseg. purified water or ethyl alcohol. The
composition may contain additives other than the inactive diluents
such as adjuvant e.g. wetting agents and suspending agents,
sweeteners, flavors, fragrance and preservatives.
[0156] The composition of the present invention may be in the form
of spraying composition, which contains one or more active
ingredients and may be prepared according to a known method.
[0157] Examples of the injectable compositions of the present
invention for parenteral administration include sterile aqueous or
non-aqueous solutions, suspensions and emulsions.
[0158] Diluents for the aqueous solution or suspension may include,
for example, distilled water for injection, physiological saline
and Ringer's solution.
[0159] Non-aqueous diluents for solution and suspension may
include, for example, propylene glycol, polyethylene glycol,
vegetable oils such as olive oil, alcohols such as ethanol and
polysorbate. The composition may further comprise additives such as
preservatives, wetting agents, emulsifying agents, dispersing
agents and the like. They may be sterilized by filtration through,
e.g. a bacteria-retaining filter, compounding with a sterilizer, or
by means of gas or radioisotope irradiation sterilization.
[0160] The injectable composition may also be provided as a
sterilized powder composition to be dissolved in a sterilized
solvent for injection before use.
[0161] The external preparation of the invention may be any form of
the external preparations used in the fields of dermatology and
otolaryngology, which includes ointment, cream, lotion and
spray.
[0162] Another form of the composition is suppository or pessary,
which may be prepared by mixing active ingredients into a
conventional base such as cacao butter that softens at body
temperature, and nonionic surfactants having suitable softening
temperatures may be used to improve absorbability.
[0163] The term "treatment" used herein includes any means of
control such as prevention, care, relief of the condition,
attenuation of the condition, arrest of progression, etc.
[0164] The term "central nervous system disorder" used herein
includes any central nervous system disorder involved or being
associated with any type of condition and/or diseases, or caused by
ischemia, trauma, infection, inflammation, tumor, edema,
hypotension, hypoxemia, blood clot (thrombus), enzyme activation,
arterial obstruction (embolus), arteriosclerosis, metabolic
disorder, degeneration, aging, drugs, medications or surgical
procedures.
[0165] Examples of "central nervous system disorder" include, but
not limited to, cerebrovascular disorders such as stroke and
cerebral infarction (e.g., cerebral thrombosis, cerebral embolism,
lacunar cerebral infarction, asymptomatic cerebral'infarction);
vasospasm due to intracerebral hemorrhage or subarachnoid
hemorrhage; cerebrovascular dementia; neuronal disorders such as
Alzheimer disease, Parkinson's disease, Huntington's chorea,
dementia, Pick disease, spino-cerebellar degeneration, chorea, AIDS
encephalopathy, hepatic encephalopathy, amyotrophic lateral
sclerosis, anticancer drug-induced peripheral neuropathy, diabetic
neuropathy, traumatic neurological disorder and multiple sclerosis;
cerebral edema, hypernatremic cerebral disorder and brain tumor;
ischemic diseases such as cerebral ischemia caused by vascular
disorders, transient ischemic attack (TIA), reversible ischemic
neurological deficit (RIND), cerebrovascular ischemia caused by
migraine or cocaine abuse, cerebral ischemia including epilepsy or
epileptic psychiatric symptoms, cerebral ischemia during surgical
operation (ischemic tissue injury), cerebral ischemia caused by
head injury, cerebral ischemia due to hypotension, hypoxemia or
dyspnea and cerebral ischemia due to cardiac arrest; inflammatory
cerebral disorders such as choronic relapsing multiple sclerosis,
encephalomyelitis, meningitis, traumatic brain injury; neonatal
asphyxia and secondary complications of these diseases.
[0166] According to the present invention, the compounds used
herein have a significant effect on recovery of barrier function of
cerebrovascular endothelial cells, especially blood brain barrier,
so it is also useful for protecting cerebrovascular endothelial
cells.
[0167] The pharmaceutical composition of the present invention may
further contain other pharmacological ingredients as far as they do
not contradict the purpose of the present invention.
[0168] The present formulations may contain a single active
ingredient or a combination of two or more active ingredients. In a
combination of plural active ingredients, their respective contents
may be suitably increased or decreased in consideration of their
therapeutic effects and safety.
[0169] Further, the present formulations may contain other
pharmacologically active ingredients, as far as they are not
contrary to the objects of the present invention.
[0170] The present invention will be described in detail with
reference to the following examples, which, however, are not
intended to limit the scope of the present invention.
Example 1
Method
[0171] Four-week-old male ddY mice were housed in aluminum cages in
an animal room controlled for temperature (24.+-.3.degree. C.),
relative humidity (55.+-.10%), ventilation rate (.about.12
times/hour) and light-dark cycle (fluorescent lighting: 8:00 to
20:00) for at least 7 days. The animals were allowed free access to
pellet diet and tap water from water bottles. Healthy animals
without abnormalities in general signs were used in this study.
[0172] 11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1
(hereinafter, "Compound A") was dissolved in a vehicle (physiologic
saline containing 0.01% polysorbate 80 and 0.5% ethanol), and was
administered subcutaneously to the animals. The control group
received an equal amount of the vehicle in the same manner.
The animals were decapitated at 30 minutes after the
administration, and the persistent time of gasping movements was
measured.
Results
[0173] As shown in Table 1, Compound A at 10, 30, 100 and 300
.mu.g/kg produced a dose-dependent prolongation of the persistent
time of gasping movement after decapitation. The results indicate
that Compound A has a neuroprotective activity and that Compound A
is useful for the treatment of ischemic disease.
TABLE-US-00001 TABLE 1 Effects of Compound A on Persistent Time of
Gasping Movements after Decapitation in Mice Dose Persistent Time
of Level Dose No. of Gasping Movements Group (.mu.g/kg) Route
Animals (sec, Mean .+-. SE) Control 0 s.c. 10 20.7 .+-. 0.6
(Vehicle) Compound A 10 s.c. 10 21.7 .+-. 0.6 Compound A 30 s.c. 10
22.0 .+-. 0.4 Compound A 100 s.c. 10 23.2 .+-. 0.8* Compound A 300
s.c. 10 23.6 .+-. 0.6** s.c.: subcutaneous, **p < 0.01, *p <
0.05 compared to vehicle-treated control group (Dunnett's multiple
comparison test).
Example 2
Method
[0174] Four-week-old male ddY mice were housed in aluminum cages in
an animal room controlled for temperature (24.+-.3.degree. C.),
relative humidity (55.+-.10%), ventilation rate (.about.12
times/hour) and light-dark cycle (fluorescent lighting: 8:00 to
20:00) for at least 7 days. The animals were allowed free access to
pellet diet and tap water from water bottles. Healthy animals
without abnormalities in general signs were used in this study. The
animals were fasted for 20 hours or longer with free access to
water before use.
[0175] Compound A and
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1 methyl
ester (hereinafter, "Compound B") were dissolved in a vehicle
(physiologic saline containing 0.01% polysorbate 80 and 0.5%
ethanol), and was administered orally to the animals. The control
group received an equal amount of the vehicle in the same
manner.
[0176] The animals were decapitated at 30 minutes after the
administration, and the persistent time of gasping movements was
measured.
<Results>
[0177] As shown in Table 2, oral administration of Compound A and
Compound B at 100, 300 and 1000 .mu.g/kg produced a dose-dependent
prolongation of the persistent time of gasping movement after
decapitation. The results indicate that Compound A and Compound B
have a neuroprotective activity by oral administration and that
Compound A and Compound B are useful for the treatment of ischemic
disease.
TABLE-US-00002 TABLE 2 Effects of Oral Administration of Compound A
and B on Persistent Time of Gasping Movements after Decapitation in
Mice Dose Persistent Time of Level Dose No. of Gasping Movements
Group (.mu.g/kg) Route Animals (sec, Mean .+-. SE) Control 0 p.o.
10 17.6 .+-. 0.4 (Vehicle) Compound A 100 p.o. 10 18.8 .+-. 0.5
Compound A 300 p.o. 10 18.9 .+-. 0.3 Compound A 1000 p.o. 10 20.2
.+-. 0.6** Compound B 100 p.o. 10 17.6 .+-. 0.5 Compound B 300 p.o.
10 19.1 .+-. 0.5 Compound B 1000 p.o. 10 19.1 .+-. 0.4 p.o.: per
os, **p < 0.01, *p < 0.05 compared to vehicle-treated control
group (Dunnett's multiple comparison test).
Example 3
Method
[0178] Four-week-old male ddY mice were housed in aluminum cages in
an animal room controlled for temperature (24.+-.3.degree. C.),
relative humidity (55.+-.10%), ventilation rate (.about.12
times/hour) and light-dark cycle (fluorescent lighting: 8:00 to
20:00) for at least 7 days. The animals were allowed free access to
pellet diet and tap water from water bottles. Healthy animals
without abnormalities in general signs were used in this study.
[0179] 11-deoxy-13,14-dihydro-16,16-difluoro-PGE.sub.1
(hereinafter, "Compound C") was dissolved in a vehicle (physiologic
saline containing 0.01% polysorbate 80 and 0.5% ethanol), and was
administered subcutaneously to the animals. The control group
received an equal amount of the vehicle in the same manner.
[0180] The animals were decapitated at 30 minutes after the
administration, and the persistent time of gasping movements was
measured.
<Results>
[0181] As shown in Table 3, Compound C at 300 .mu.g/kg produced a
significant prolongation of the persistent time of gasping movement
after decapitation. The results indicate that Compound C has a
neuroprotective activity.
TABLE-US-00003 TABLE 3 Effect of Compound C on Persistent Time of
Gasping Movements after Decapitation in Mice Dose Persistent Time
of Level Dose No. of Gasping Movements Group (.mu.g/kg) Route
Animals (sec, Mean .+-. SE) Control 0 s.c. 10 21.9 .+-. 0.5
(Vehicle) Compound C 300 s.c. 10 25.2 .+-. 0.7** s.c.:
subcutaneous, **p < 0.01 compared to vehicle-treated control
group.
Example 4
Method
[0182] Seven-week-old Crj: CD (SD) male rats were housed in
polymethylpentene cages in an animal room controlled for room
temperature (22-26.degree. C.), relative humidity (47-60%),
ventilation rate (10-20 times/hour) and light-dark cycle (lighting:
7:00 to 19:00) for at least 6 days. The animals were allowed free
access to pellet diet and water from water bottles. Animals judged
to be in good health were used in this study.
[0183] Rats were anesthetized by inhalation of a gas mixture of 2%
isoflurane and N.sub.2O:O.sub.2 (=7:3), stabilized in the supine
position, and maintained in the anesthetized state by inhalation of
the above gas mixture. The animals were monitored for rectal
temperature using a temperature probe during the period of the
surgical operation. When a fall in body temperature was observed,
an incandescent lamp was used to maintain the temperature at around
37.degree. C. The right common carotid artery, external carotid
artery, and internal carotid artery were exposed for occluding the
middle cerebral artery (hereafter, MCA). The right common carotid
artery and the external carotid artery were ligatured using sutures
(5-0), and a 19 mm-long segment of No. 4-0 nylon suture which were
precoated with silicone was inserted into the MCA through the
bifurcation of the external and internal carotid arteries to
occlude the MCA. At 2 hours after the MCA occlusion, the suture was
removed and the blood flow in the MCA was restored.
[0184] Compound A was dissolved in a vehicle (physiological saline
containing 1% polysorbate 80), and was administered intravenously
to the animals at a volume of 2 ml/kg immediately after the MCA
occlusion-reperfusion and minutes after the MCA
occlusion-reperfusion. The control group received an equal volume
of the vehicle in the same manner.
[0185] At 24 hours after MCA occlusion, the animals were
decapitated and the brains were immediately isolated. Using a
tissue chopper (Micro-3D; The Mickle Laboratory Engineering Co.,
Ltd.), sequential brain sections 2 mm in thickness were prepared.
The brain tissue sections were positioned following the brain atlas
of Paxinos and Watson to include the coronal plane at 4 mm anterior
to the bregma, at 2 mm anterior to the bregma, at the bregma, at 2
mm posterior to the bregma, at 4 mm posterior to the bregma, and at
6 mm posterior to the bregma. The brain sections were stained in 1%
TTC solution and photographed. Graphic analysis (Adobe
Photoshop.TM., version 3.0 J; Adobe Systems Incorporated, Color
Count 0.3b; K&M Software Corporation) was applied to the
photographs, and the infarct area was measured. Based on these
results, the infarct volume (4 mm anterior to the bregma--6 mm
posterior to the bregma) was calculated using the following
formula.
V = 2 ( a + b ) / 2 + 2 ( b + c ) / 2 + 2 ( c + d ) / 2 + 2 ( d + e
) / 2 + 2 ( e + f ) / 2 = a + 2 ( b + c + d + e ) + f ##EQU00001##
[0186] V: infarct volume [0187] a: infarct area at the
cross-section 4 mm before the bregma [0188] b: infarct area at the
cross-section mm before the bregma [0189] c: infarct area at the
cross-section just at the bregma [0190] d: infarct area at the
cross-section 2 mm behind the bregma [0191] e: infarct area at the
cross-section 4 mm behind the bregma [0192] f: infarct area at the
cross-section 6 mm behind the bregma
<Results>
[0193] As shown in Table 4, Compound A at 0.05 and 0.5 mg/kg
significantly reduced the cerebral infarct volume after ischemia in
a dose-dependent manner compared with that in the vehicle group.
The results indicate that Compound A is useful for the treatment of
cerebrovascular disorders such as cerebral infarct.
TABLE-US-00004 TABLE 4 Effects of Compound A on cerebral infarct
volume after transient focal cerebral ischemia in rats Dose Infarct
volume Group mg/kg n mm.sup.3 Control -- 10 280.8 .+-. 16.2
(Vehicle) Compound A 0.05 10 208.2 .+-. 22.2* Compound A 0.5 10
172.9 .+-. 25.5**
[0194] Brain was removed at 24 hours after MCA occlusion. Each
value represents the mean.+-.S.E. of 10 rats. Compounds were
administered intravenously immediately after MCA
occlusion--reperfusion and 30 minutes after MCA
occlusion-reperfusion. * P<0.05, P<0.01; Significant
difference from vehicle group and Compound A group (Dunnett's
multiple comparison test)
Example 5
Method
[0195] Alzheimer's disease model animals were prepared by bilateral
ibotenic acid lesions of basal ganglia in rats. Briefly, rats were
anesthetized with pentobarbital sodium and placed in a small animal
stereotaxic apparatus. Bilateral infusions of 5 .mu.g/0.5 .mu.L of
ibotenic acid into the basal ganglia were made at a rate of 0.1
.mu.L/min via a syringe pump and a stainless steel cannula (outer
diameter: 0.5 mm). Stereotaxic coordinates were as follows: -0.8 mm
posterior from bregma, 2.6 mm lateral (both sides) from midline,
and 7.4 mm depth from the bone surface. Animals in sham group
received only anesthesia. Animals were then housed with free access
to food and water for the rest of the study.
[0196] Compound A was orally administered for 14 days after surgery
to the model animals. Control group received the same amount of the
vehicle.
[0197] Morris water maze test was performed to evaluate the effect
of test compound. The water maze was a circular pool (painted gray,
1.48 m in diameter, 0.33 m high). The pool contained water that was
maintained at a temperature of 17-18.degree. C. During testing in
the water maze, a platform, 12 cm in diameter, was located 2 cm
below the water in one of four locations (zone 4) in the pool,
approximately 38 cm from the sidewall. A light bulb was placed
around the pool as a cue external to the maze. The animals received
2 trials per day from 10 days after the initiation of the
administration with Compound A or the vehicle. The rats were
trained to locate the hidden escape platform, which remained in a
fixed location throughout testing. Trials lasted a maximum of 90
sec. The latency to find the submerged platform was recorded and
used as a measure of acquisition of the task. The animals were
tested in this way for 4 days (total 8 trials), and then they
received a probe trial on the 5th day. For the probe trial, the
platform was removed from the pool and then the animal was released
from the quadrant opposite to where the platform would have been
located. The length of the trial was 90 sec, after which the rat
was removed from the pool. The time the rat spent searching for the
platform in the training quadrant (zone 4); i.e., the previous
location of the platform was recorded and used as an index of
memory.
<Results>
[0198] As shown in table 5 and 6, vehicle group showed severely
impaired spatial cognition. Treatment with Compound A produced
significant reversals of the deficit in learning and memory. These
results suggest that Compound A is useful for the treatment of
neuronal disorders such as Alzheimer's disease.
TABLE-US-00005 TABLE 5 Effect of Compound A on goal latency in
Morris water maze learning test. Dose Goal latency (8.sup.th trial)
Group mg/kg n mean .+-. SE, sec Sham 0 10 24.6 .+-. 2.7 Vehicle 0
10 90.0 .+-. 0.0.sup.## Compound A 1 10 51.5 .+-. 13.7** .sup.##p
< 0.01 compared with sham group, **p < 0.01 compared with
vehicle group
TABLE-US-00006 TABLE 6 Effect of Compound A on time spent in
quadrant (zone 4) where previous location of the platform in Morris
water maze learning test. Dose Time spent in zone 4 Group mg/kg n
mean .+-. SE, sec Sham 0 10 24.5 .+-. 2.0 Vehicle 0 10 12.2 .+-.
1.5.sup.## Compound A 1 10 20.8 .+-. 3.6* .sup.##p < 0.01
compared with sham group, *p < 0.05 compared with vehicle
group
Example 6
Method
[0199] Human vascular endothelial cell cultures were brought to
confluence, as measured by transendothelial electrical resistance
(TEER). The cell cultures were then deprived of oxygen for 30
minutes by incubation in a nitrogen atmosphere. The cells were then
either treated with 0.1% DMSO or with 5 nM Compound A with 0.1%
DMSO final.
<Results>
[0200] As shown FIG. 1, the DMSO-treated cells showed very little
recovery of TEER. The Compound A-treated cells showed immediate
recovery of TEER.
[0201] The results demonstrate that TEER, a measured barrier
function of endothelial cells, recovers rapidly from damage after
Compound A-treatment.
Example 7
Method
[0202] Human microvascular endothelial cells (adult) (HMVEC-AD)
were grown to confluence. The cells were then treated for 30
minutes with a nitrogen atmosphere and returned to normal oxygen.
ATP levels were monitored at the indicated time points using a
luciferin-luciferase assay system (ATPlite, Perkin Elmer).
<Results>
[0203] As shown in FIG. 2, ATP levels decreased when the cells were
exposed to a nitrogen atmosphere for 30 minutes. ATP levels
returned more quickly in cells treated with 5 nM Compound A
compared to cells treated with 0.01% DMSO alone.
[0204] The results indicate that the Compound A is useful for the
treatment of central nervous system disorders.
Synthesis Example 1
##STR00011##
[0205] Synthesis of 16,16-difluoro-PGA.sub.1 benzyl Ester (2)
[0206] 16,16-Difluoro-PGE.sub.1 benzyl ester (1) (457.8 mg, 0.95
mmol) was dissolved in acetic acid (13.7 mL, 0.24 mol), and the
solution was stirred at 80.degree. C. for 18 hours. The reaction
mixture was cooled to room temperature. 10 mL of toluene was added
to the solution and concentrated under reduced pressure. This
operation was repeated five times to removed acetic acid. The
residue was purified by silica gel column chromatography (silica
gel: FL60D (70 g), Fuji Silysia, hexane/ethyl acetate (2:1)) to
obtain compound (2) as yellow oil. Yield: 391.6 mg (88.9%).
Synthesis of 11-deoxy-13,14-dihydro-16,16-difluoro-PGE.sub.1 (3)
16,16-Difluoro-PGA.sub.1 benzyl ester (compound (2)) (382.5 mg,
0.83 mmol) was hydrogenated in ethyl acetate (10 mL) under the
presence of 10% palladium-carbon (57.4 mg, wet with 50% w/w of
water) at room temperature, at atmospheric pressure for 2 hours.
The reaction mixture was filtered through a Celite pad, the filter
cake was washed with ethyl acetate, and then the filtrate was
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (silica gel BW-300SP (50 g, wet
with 15% w/w of water), Fuji Silysia, hexane/ethyl acetate (1:1))
to obtain crude compound (3) (298.5 mg, 95.7%).
[0207] The crude compound (3) was combined with another lot of the
crude compound. And then, totally about 350 mg of the crude
compound was purified by preparative HPLC (YMC-Pack D-SIL-5-06
20.times.250 mm, hexane/2-propanol/acetic acid (250:5:1), 20
mL/min) to obtain compound (3) as colorless oil. Yield: 297.3 mg
(HPLC purification recovery: 83.5%). .sup.1H-NMR (200 MHz,
CDCl.sub.3) .delta.
[0208] 0.94 (3H, t, J=7.1 Hz), 1.22-2.29 (28H, m), 2.34 (2H, t,
J=7.3 Hz), 3.65-3.81 (1H, m)
[0209] .sup.13C-NMR (50 MHz, CDCl.sub.3) .delta.
[0210] 13.70, 22.40, 23.25, 24.32, 26.28, 26.63), 27.18, 27.58,
28.49, 29.09, 30.39, 31.77 (t, J=24.4 Hz), 33.67, 37.63, 41.05,
54.76, 72.73 (t, J=29.0 Hz), 124.09 (t, J=244.3 Hz), 179.07,
220.79.
Synthesis Example 2
##STR00012##
[0212] According to the similar manner described in Synthesis
Example 1, 11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1
isopropyl ester (Compound (6)) was obtained as colorless oil by the
above two-step reaction. Yield: 0.285 g (1.sup.st step: 96.2%,
2.sup.nd step: 97.6%, HPLC purification: recovery 81.0%).
.sup.1H-NMR (200 MH2, CDCl.sub.3) and .sup.13C-NMR (50 MHz,
CDCl.sub.3) of the Compound (6) are shown in FIGS. 3 and 4
respectively.
Synthesis Example 3
##STR00013##
[0214] According to the similar manner described in Synthesis
Example
1,2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-dif-
luoro-PGE.sub.1 isopropyl ester (Compound (9)) was obtained as
colorless oil. Yield: 0.402 g (1.sup.st step: 94.9%, 2.sup.nd step:
92.2%, HPLC purification: recovery 83.1%). .sup.1H-NMR (200 MHz,
CDCl.sub.3) and .sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound
(9) are shown in FIGS. 5 and 6 respectively.
Synthesis Example 4
##STR00014##
[0216] According to the similar manner described in Synthesis
Example 1,
2-decarboxy-2-(2-carboxyethyl)-11-deoxy-13,14-dihydro-15-keto-16,16-diflu-
oro-PGE.sub.1 (Compound (12)) was obtained as colorless oil. Yield:
0.696 g (1' step: 95.6%, 2.sup.nd step: 99.3%, HPLC purification:
recovery: 87.4%). .sup.1H-NMR (200 MHz, CDCl.sub.3) and
.sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound (12) are shown in
FIGS. 7 and 8 respectively.
Synthesis Example 5
##STR00015##
[0218] According to the similar manner described in Synthesis
Example 1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1
isopropyl ester (Compound (15)) was obtained as colorless oil.
Yield: 0.271 g (1.sup.st step: 91.4%, 2.sup.nd step: 97.3%, HPLC
purification: recovery: 79.0%). .sup.1H-NMR (200 MHz, CDCl.sub.3)
and .sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound (15) are
shown in FIGS. 9 and 10 respectively.
Synthesis Example 6
##STR00016##
[0220] According to the similar manner described in Synthesis
Example 1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-methyl-PGE.sub.1
(Compound (18)) was obtained as colorless oil. Yield: 0.637 g
(1.sup.st step: 93.3%, 2.sup.nd step: 96.6%, HPLC purification:
recovery: 73.9%). .sup.1H-NMR (200 MHz, CDCl.sub.3) and
.sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound (18) are shown in
FIGS. 11 and 12 respectively.
Synthesis Example 7
##STR00017##
[0222] According to the similar manner described in Synthesis
Example 1,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.1
(Compound (21)) was obtained as colorless oil. Yield: 0.401 g
(1.sup.st step: 90.6%, 2.sup.nd step: 92.7%, HPLC purification:
recovery: 29.2%). .sup.1H-NMR (200 MHz, CDCl.sub.3) and
.sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound (21) are shown in
FIGS. 13 and 14 respectively.
Synthesis Example 8
##STR00018##
[0224] 11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGE.sub.1
methyl ester (Compound (23)) was obtained as colorless oil by
esterification of compound (22) with diazomethane. Yield: 0.860 g
(72.9%, after purification by silica gel column chromatography).
.sup.1H-NMR (200 MHz, CDCl.sub.3) and .sup.13C-NMR (50 MHz,
CDCl.sub.3) of the Compound (23) were shown in FIGS. 15 and 16.
Synthesis Example 9
##STR00019##
[0226] Compound (24) (0.67 g, 1.66 mmol) was dissolved in DMF (13
mL), and added K.sub.2CO.sub.3 (460.1 mg, 3.33 mmol) and isopropyl
iodide (831 .mu.L, 8.32 mmol). The solution was stirred at room
temperature for 2 hours. The reaction mixture was cooled with ice,
added water (10 mL) and brine, and extracted with ethyl acetate (30
mL). The organic layer was washed with brine (10 mL), dried with
anhydrous magnesium sulfate, and then concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography (silica gel FL60D (50 g), Fuji Silysia, hexane/ethyl
acetate (5:1)) to obtain crude
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-20-ethyl-PGE.sub.1
isopropyl ester (compound (25)) (0.70 g, 94.6%). The crude compound
(25) was purified by preparative HPLC to obtain compound (25) as
colorless oil. Yield 245.8 mg (35.1%). .sup.1H-NMR (200 MHz,
CDCl.sub.3) and .sup.13C-NMR (50 MHz, CDCl.sub.3) for the Compound
(25) are shown in FIGS. 17 and 18 respectively.
Synthesis Example 10
##STR00020## ##STR00021##
[0228] Compound (26) (8.71 g, 20.2 mmol) was dissolved in
1,2-dichloroethane (70 mL) and added 1,1'-Thiocarbonyldiimidazole
(5.41 g, 30.3 mmol). The solution was stirred at 70.degree. C. for
an hour. The reaction mixture was cooled to room temperature, and
then concentrated under reduced pressure. The residue was purified
by silica gel column chromatography (silica gel BW-300SP (650 g),
Fuji Silysia, hexane/ethyl acetate (1:1)) to obtain compound (27)
as light yellow oil (10.61 g, 97.0%).
[0229] Bu.sub.3SnH (11.21 g, 38.5 mmol) was dissolved in toluene
(224 mL), and refluxed by heating. The solution of Compound (27)
(10.41 g, 19.2 mmol) in toluene (208 mL) was dropped to the
reaction mixture at a reflux temperature for 70 minutes. And then,
the reaction mixture was cooled to room temperature, concentrated
under reduced pressure to obtain crude compound (28) as light
yellow oil.
[0230] The crude compound (28) (19.2 mmol) was dissolved in THF (52
mL) and TBAF solution (1.0M in THF, 38.5 mL, 38.5 mmol) was dropped
for 10 minutes. After an hour, TBAF solution (1.0M in THF, 19.2 ml,
19.2 mmol) was dropped to the solution. After stirring for total
3.5 hours, the reaction mixture was concentrated under reduced
pressure. The residue was purified by silica gel column
chromatography (silica gel BW-300SP (1,000 g), Fuji Silysia,
hexane/ethyl acetate (1:1)) to obtain compound (29) as yellow oil
(4.01 g, 69.3%).
[0231] Compound (31) was obtained from compound (29) by Swern
oxidation and introduction of .omega.-chain.
[0232] Compound (31) (807.4 mg, 1.88 mmol) was hydrogenated in
ethyl acetate (8 mL) under the presence of 10% palladium-carbon at
room temperature for 2 hours. The reaction mixture was filtered
through a Celite pad, and the filtrate was concentrated under
reduced pressure to obtain crude compound (32) as the light brown
oil.
[0233] The crude compound (32) (1.88 mmol) was dissolved in EtOH (8
mL). 1N--NaOH solution (7.4 mL, 7.4 mol) was dropped to the
solution at room temperature for 10 minutes. The reaction mixture
was stirred at room temperature for 10 hours, and then cooled with
ice. 1N--HCl (7.1 mL) was dropped to the reaction mixture to adjust
pH around 3-4. Then the reaction mixture was extracted with TBME
(30 mL). The organic layer was washed with water (10 mL) and brine
(10 mL), dried with anhydrous magnesium sulfate, and then
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (silica gel 15% water including
FL-60D (80 g), Fuji Silysia, hexane/ethyl acetate (2:1)) to obtain
compound (33) as light yellow oil (481.4 mg, 68.8%).
[0234] According to the similar manner described in Synthesis
Example 9,
11-deoxy-13,14-dihydro-15-keto-16,16-difluoro-PGF.sub.1.alpha.
isopropyl ester (compound (34)) was obtained from compound (33) as
colorless oil. Yield: 166.6 mg (reaction step 91.9%: HPLC
purification: recovery: 55.4%). .sup.1H-NMR (200 MHz, CDCl.sub.3)
and .sup.13C-NMR (50 MHz, CDCl.sub.3) of the Compound (34) are
shown in FIGS. 19 and 20 respectively.
* * * * *