U.S. patent application number 09/900021 was filed with the patent office on 2002-02-21 for treatment of ocular hypertension and glaucoma.
This patent application is currently assigned to R-TECH UENO, LTD.. Invention is credited to Ueno, Ryuji.
Application Number | 20020022644 09/900021 |
Document ID | / |
Family ID | 27414967 |
Filed Date | 2002-02-21 |
United States Patent
Application |
20020022644 |
Kind Code |
A1 |
Ueno, Ryuji |
February 21, 2002 |
Treatment of ocular hypertension and glaucoma
Abstract
Disclosed is treatment of ocular hypertension and glaucoma by
long-term therapy with a prostaglandin related compound for
eliminating or reducing potential iridic pigmentation. Composition
useful for the treatment, and use of the prostaglandin related
compound for producing the composition are also disclosed.
Inventors: |
Ueno, Ryuji; (Potomac,
MD) |
Correspondence
Address: |
SUGHRUE MION ZINN MACPEAK & SEAS, PLLC
2100 Pennsylvania Avenue, NW
Washington
DC
20037-3213
US
|
Assignee: |
R-TECH UENO, LTD.
|
Family ID: |
27414967 |
Appl. No.: |
09/900021 |
Filed: |
July 9, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09900021 |
Jul 9, 2001 |
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09817046 |
Mar 27, 2001 |
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09817046 |
Mar 27, 2001 |
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09730830 |
Dec 7, 2000 |
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09730830 |
Dec 7, 2000 |
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09527573 |
Mar 16, 2000 |
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Current U.S.
Class: |
514/330 ;
514/461; 514/560 |
Current CPC
Class: |
A61K 31/5575 20130101;
Y10S 514/913 20130101; A61K 31/5585 20130101; A61K 31/557
20130101 |
Class at
Publication: |
514/330 ;
514/461; 514/560 |
International
Class: |
A61K 031/5585; A61K
031/5575 |
Claims
What is claimed:
1. A process for the long-term treatment or prophylactic management
of ocular hypertension or glaucoma in a human patient by topical
ocular administration of a therapeutically effective amount of a
prostaglandin related compound, the improvement to eliminate or
reduce potential pigmentation of the iris of a treated eye
occurring by long-term topical ocular application of a
prostaglandin related compound.
2. A process for the long-term treatment or prophylactic management
of ocular hypertension or glaucoma in a human patient by topical
ocular administration of a therapeutically effective amount of a
prostaglandin related compound, the improvement to eliminate or
reduce potential pigmentation of the iris of a treated eye
occurring by long-term topical ocular application of a
prostaglandin related compound, wherein the compound used has the
formula (I) 8wherein W.sub.1, W.sub.2 and W.sub.3 are carbon or
oxygen atoms, L, M and N are hydrogen atom, hydroxy, halogen atom,
lower alkyl, lower alkoxy, hydroxy(lower)alkyl, or oxo, wherein at
least one of L and M is a group other than hydrogen, and the
five-membered ring may have at least one double bond; A is
--CH.sub.2OH, --COCH.sub.2OH, --COOH or a functional derivative
thereof; B is single bond, --CH.sub.2--, --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--; R.sub.1 is a
saturated or unsaturated bivalent lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen, an alkyl group, hydroxy, oxo, aryl or heterocyclic group;
and Ra is a saturated or unsaturated lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen atom, oxo, hydroxy, lower alkyl, lower alkoxy, lower
alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl,
aryloxy, heterocyclic group or heterocyclic-oxy group;
cyclo(lower)alkyl; cyclo(lower)alkyloxy; aryl; aryloxy;
heterocyclic group; or heterocyclic-oxy group.
3. The process of claim 2 wherein the functional derivative of A is
salts, ethers, esters or amides.
4. The process of claim 3 wherein the amide of A is 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.
5. The process of claim 3 wherein the compound used is a
13,14-dihydro-15-keto-20 ethyl PGF.sub.2.alpha. isopropyl
ester.
6. The process of claim 3 wherein the compound used is 15-keto
latanoprost.
7. The process of claim 3 wherein the compound used is
15-keto-17-phenyl-18,19,20-trinor PGF.sub.2.alpha.
N-ethylamide.
8. The process of claim 3 wherein the compound used is
13,14,dihydro-15-keto 17-phenyl-18,1920-trinor PGF.sub.2.alpha.
N-ethylamide.
9. The process of claim 3 wherein the compound used is
15-keto-16-(3-trifluoromethyl phenoxy)-17,18,19,20-tetranor
PGF.sub.2.alpha. isopropyl ester.
10. The process of claim 3 wherein the compound used is
13,14-dihydro-15-keto-16-(3-trifluoromethyl
phenoxy)-17,18,19,20-tetranor PGF.sub.2.alpha. isopropyl ester.
11. The process of any one of claims 5-10 wherein the compound used
is administered at least once a day for at least six months.
12. The process of claim 11 wherein the compound used is
administered twice daily for at least six months.
13. The process of claim 1 wherein the human patient is a
Caucasian.
14. The process of claim 1 wherein the compound used contains a
15-keto substituent and an omega chain containing a phenyl
ring.
15. A process for the long-term treatment or prophylactic
management of ocular hypertension or glaucoma in a human patient by
topical ocular administration of a therapeutically effective amount
of a prostaglandin related compound, the improvement to eliminate
or reduce potential pigmentation of the iris of a treated eye
occurring by long-term topical ocular application of a
prostaglandin related compound, wherein the compound used has the
formula (III) 9wherein W.sub.1, W.sub.2 and W.sub.3 are carbon or
oxygen atoms, L, M and N are hydrogen atom, hydroxy, halogen atom,
lower alkyl, lower alkoxy, hydroxy(lower)alkyl, or oxo, wherein at
least one of L and M is a group other than hydrogen, and the
five-membered ring may have at least one double bond; A is
--CH.sub.2OH, --COCH.sub.2OH, --COOH or a functional derivative
thereof; B is single bond, --CH.sub.2--, --CH.sub.2--CH.sub.2--,
--CH.ident.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--; R.sub.1 is
a saturated or unsaturated bivalent lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen, an alkyl group, hydroxy, oxo, aryl or heterocyclic group;
Z is 10wherein R.sub.4 and R.sub.5 are hydrogen atom, hydroxy,
halogen atom, lower alkyl, lower alkoxy or hydroxy(lower)alkyl,
wherein R.sub.4 and R.sub.5 are not hydroxy, lower alkoxy and/or
hydroxy (lower) alkyl at the same time; and Ra' comprises (1) a
saturated or unsaturated C.sub.3 to C.sub.5 straight chain
aliphatic hydrocarbon beyond carbon atom number 15 substituted by
cyclo (lower)alkyl, cyclo (lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group at its terminus or (2)
a saturated or unsaturated at least C.sub.6 straight chain
aliphatic hydrocarbon residue beyond carbon atom number 15, which
is unsubstituted or substituted with halogen atom, oxo, hydroxy,
lower alkyl, lower alkoxy, lower alkanoyloxy, cyclo(lower)alkyl,
cyclo(lower)alkyloxy, aryl, aryloxy, heterocyclic group or
heterocyclic-oxy group.
16. The process of claim 15 wherein the functional derivative of A
is salts, ethers, esters or amides.
17. The process of claim 16 wherein the amide of A is 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.
18. The process of claim 15 wherein the compound used contains a
straight chain beyond carbon atom number 15 of at least 6 carbon
atoms.
19. The process of claim 18 wherein the compound used is a
docosanoid.
20. The process of claim 15 wherein the compound used contains a
straight chain beyond carbon atom number 15 of at least 3 carbon
atoms substituted by cyclo(lower)alkyl, cyclo(lower) alkyloxy,
aryl, aryloxy, heterocyclic group ar heterocyclic-oxy group at the
terminus of the omega chain.
21. The process of claim 20 wherein the compound used is
substituted by is phenyl at the terminus of the omega chain.
22. The process of claim 19 wherein the compound used is
administered at least once a day for at least six months.
23. The process of claim 22 wherein the compound used is
administered twice daily for at least six months.
24. The process of claim 2 or claim 15 wherein the human patient is
a Caucasian.
25. A process for the long term treatment or prophylactic
management of ocular hypertension or glaucoma in a human patient by
topical ocular administration in a human patient of a
therapeutically effective amount of a prostaglandin related
compound, the improvement to eliminate or reduce potential
pigmentation of the iris of a treated eye occurring by long-term
topical ocular application of a prostaglandin related compound,
wherein the compound used has the formula (IV) 11wherein W.sub.1,
W.sub.2, and W.sub.3 are carbon or oxygen atoms, L, M and N are
hydrogen atom, hydroxy, halogen atom, lower alkyl, lower alkoxy,
hydroxy(lower)alkyl, or oxo, wherein at least one of L and M is a
group other than hydrogen, and the five-membered ring may have at
least one double bond; R' and R" is hydrogen atom, lower alkyl,
aryl, alkyl- or aryl-sulfonyl, lower alkenyl and lower alkynyl; B
is single bond, --CH.sub.2--, --CH.sub.2--CH.sub.2--,
--CH.dbd.CH--, --C.dbd.C--, --CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--, --CH.sub.2CH.dbd.CH--,
--C.ident.--C--CH.sub.2--, or CH.sub.2--C.ident.--C--; R.sub.1 is a
saturated or unsaturated bivalent lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen, an alkyl group, hydroxy, oxo, aryl or heterocyclic group;
Z is 12wherein R.sub.4 and R.sub.5 are hydrogen atom, hydroxy,
halogen atom, lower alkyl, lower alkoxy or hydroxy(lower)alkyl,
whrein R.sub.4 and R.sub.5 are not hydroxy, lower alkoxy and/or
hydroxy(lower)alkyl at the same time; and Ra is a saturated or
unsaturated lower or medium aliphatic hydrocarbon residue, which is
unsubstituted or substituted with halogen atom, oxo, hydroxy, lower
alkyl, lower alkoxy, lowr alkanoyloxy, cyclo(lower)alkyl,
cyclo(lower))alkyloxy, aryl, aryloxy, heterocyclic group or
hetrocyclic-oxy group; cyclo(lower)alkyl; cyclo(lower)alkyloxy;
aryl; aryloxy; heterocyclic group; heterocyclic-oxy group.
26. The process of claim 25 wherein the compound used is
17-phenyl-18,19,20-trinor PGF.sub.2.alpha. N-ethylamide.
27. The process of claim 25 wherein the compound used is
administered at least once a day for at least six months.
28. The process of claim 25 wherein the compound used is
administered twice daily for at least six months.
29. The process of claim 28 wherein the human patient is a
Caucasian.
Description
[0001] This application is a continuation-in-part application of
U.S. Ser. No. 09/817,046 filed Mar. 27, 2001, in turn a
continuation-in-part application of U.S. Ser. No. 09/730,830 filed
Dec. 7, 2000, in turn a continuation-in-part application of U.S.
Ser. No. 09/527,573 filed Mar. 16, 2000, now abandoned.
DESCRIPTION
BACKGROUND OF THE INVENTION
[0002] The present invention relates to the long-term treatment and
prophylactic management of intraocular pressure in human patients.
Further, the present invention relates to a composition useful for
said treatment and management. Further more, the present invention
relates to use of a specific compound for manufacturing said
pharmaceutical composition. More specifically, the present
invention relates to the long term management of hypertension or
glaucoma in the eyes of human patients, without causing
pigmentation or with causing comparatively minimal pigmentation of
the iris, by periodic topical ocular application of a prostaglandin
related compound.
[0003] Prostaglandins (hereinafter, referred to as PG(s)) are
members of a class of organic carboxylic acids, which are contained
in tissues or organs of human and most other animals, 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): 1
[0004] On the other hand, some of the synthetic analogues of
primary PGs have a modified skeleton. The primary PGs are
classified to 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:
[0005] Subscript 1: 13,14-unsaturated-15-OH
[0006] Subscript 2: 5,6- and 13,14-diunsaturated-15-OH
[0007] Subscript 3: 5,6-, 13,14-, and
17,18-triunsaturated-15-OH.
[0008] Further, the PGFs are classified, according to the
configuration of the hydroxyl group at the 9-position, into .alpha.
type (the hydroxyl group is of an .alpha.-configuration) and .beta.
type (the hydroxyl group is of a .beta.-configuration).
[0009] In addition, some 15-keto (i.e. having an oxo group at
position 15 in place of the hydroxy group) prostaglandins and
13,14-dihydro-15-keto-p- rostaglandins are known as substances
naturally produced by enzymatic reactions during in vivo metabolism
of primary PGs. 15-keto PGs have been disclosed in, for example,
EP-A-0281239, EP-A-0281480, EP-A-0289349, EP-A-0453127 and
EP-A-0690049. These cited references are herein incorporated by
reference.
[0010] At present, Latanoprost is available commercially in the
United States for use as a topical ocular hypotensive and an
anti-glaucoma agent. Chemically, Latanoprost is a
13,14-dihydro-17-phenyl-18,19,20-trin- or PGF.sub.2.alpha.
isopropyl ester. One side effect of Latanoprost is a brown
pigmentation of the iris found in about 10% or more of the human
patients treated with Latanoprost for about three or more months
for management of elevated intraocular pressure. Latanoprost
possesses a substantial specific binding affinity for the FP
receptor. Selen et al have reported that PGF.sub.2.alpha.-IE,
PGE.sub.2-IE and latanoprost induced increased iridial pigmentation
in cynomolgus monkeys (Survey of Ophthalmology, 41, supplement 2,
S125-S128 (1997)). ("IE" means isopropyl ester.)
[0011] Unoprostone isopropyl ophthalmic solution (Rescula.RTM.) has
been commercially available outside Europe and the United States
for topical application in the treatment of ocular hypertension and
glaucoma. Unoprostone isopropyl is a docosanoid, namely
13,14-dihydro-15-keto-20-et- hyl PGF.sub.2.alpha. isopropyl ester.
To the inventor's best knowledge, Resucla.RTM. has not been
commercially used by Caucasians in the management of ocular
hypertension or glaucoma by its periodic topical application to the
eye at least once a day for a period of at least six months, more
than one year prior to the filing date of this application.
Preliminary results regarding no iridic pigmentation from a
long-term monkey trial with Unoprostone isopropyl have been
published. Resucla.RTM. exhibits substantial absence of FP receptor
stimulatory activity.
[0012] 15-keto-latanoprost (13,14-dihydro-15-keto-17-phenyl-18, 19,
20-trinor PGF.sub.2.alpha. isopropyl ester) is a promising
candidate for use as a topical ocular hypotensive drug. Short-term
studies of its use have been reported in U.S. Pat. No.
5,321,128.
SUMMARY OF THE INVENTION
[0013] The present invention provides methods for the long-term
treatment and prophylactic management of ocular hypertension and
glaucoma in human patients without causing pigmentation or with
causing less pigmentation than latanoprost of the patient's iris,
by periodic topical administration of a prostaglandin related
compound.
[0014] The present invention also provides a composition suitable
for the long-term treatment and prophylactic management of ocular
hypertension and glaucoma in human patients by periodic topical
ocular administration, which comprises a prostaglandin related
compound as an active ingredient.
[0015] The present invention also provides use of a prostaglandin
related compound for producing a pharmaceutical composition
suitable for the long-term treatment and prophylactic management of
ocular hypertension and glaucoma in human patients by periodic
topical ocular administration.
[0016] According to the present invention, the term "prostaglandin
related compound" (hereinafter, referred as "PG related compound")
includes any of derivatives or analogs (including substituted
derivatives) of a compound having the prostanoic acid basic
structure irrespective of the configuration of the 5-membered ring,
number of double bonds in the .alpha. or .omega.-chain, presence or
absence of hydroxy and oxo groups or any other substituent, or any
other modification.
[0017] The nomenclature of the PG related compounds used herein is
based on the numbering system of the prostanoic acid skeleton
represented in the above formula (A).
[0018] The formula (A) shows a basic skeleton of 20 carbon atoms,
but the PG related compounds in the present invention are not
limited to those having a 20 carbon atom skeleton. 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 carboxy group (C-1). Similarly, when the
number of carbon atoms is decreased in the .omega.-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.
[0019] In general, each of the terms PGD, PGE and PGF represents PG
compounds having hydroxy group (s) at positions 9 and/or 11, but in
the present specification, these terms also include those PG
related compounds having substituents other than the hydroxy group
at positions 9 and/or 11 Such compounds are referred to as
9-dehydroxy-9-substituted-PG compounds or
11-dehydroxy-11-substituted-PG compounds. A PG compound having
hydrogen in place of the hydroxy group is simply named as 9- or
11-dehydroxy compound.
[0020] As stated above, the nomenclature of the PG related
compounds is based on the prostanoic acid skeleton. However, in
case the compound has a similar partial construction as a
prostaglandin, the abbreviation of "PG" may be used. Thus, a 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)-PG compound. Similarly, a PG
compound having 11 carbon atoms in the .alpha.-chain is named as
2-decarboxy-2-(4-carboxybutyl)-PG compound, and a PG compound
having 10 carbon atoms in the .omega.-chain is named as 20-ethyl-PG
compound. These compounds, however, may also be named according to
the IUPAC nomenclatures.
[0021] The PG related compounds used in the present invention may
include any of PG derivatives or analogs. Accordingly, for example,
a PG.sub.1 compound having a double bond at 13-14 position and a
hydroxy group at 15-position, a PG.sub.2 compound having another
double bond at 5-6 position, a PG.sub.3 compound having further
double bond at 17-18 position, a 15-keto-PG compound having an oxo
group in place of the hydroxy group at the 15-position, a
15-dehydroxy-PG compound having a hydrogen atom in place of the
hydroxy group at the 15-position, or the corresponding
13,14-dihydro-PG compounds wherein in each type of compound the
double bond at 13-14 position is single bond, or the corresponding
13,14-didehydro-PG compounds wherein in each type of compound the
double bond at the 13-14 position is a triple bond are included.
Moreover, examples of substituted compounds and derivatives include
a compound wherein the terminal carboxyl group in the .alpha.-chain
of the above described compound is ester, ether, amide, a
pharmaceutically acceptable salt thereof, a compound wherein the
number of carbon atoms in the .alpha.- or .omega.-chain is
decreased or increased, a compound having side chains (e.g., 1 to 3
carbon atoms) on .alpha.- or .omega.-chains, a compound having
substituent(s) such as hydroxy, halogen, lower alkyl,
hydroxy(lower)alkyl, and oxo, or double bond(s) on the
five-membered ring, a compound having substituent(s), such as
halogen, oxo, aryl and heterocyclic on the .alpha.-chain, a
compound having substituents such as halogen, oxo, hydroxy, lower
alkoxy, lower alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy,
aryl, aryloxy, heterocyclic group and heterocyclic-oxy group on the
.omega.-chain, and a compound having substituent such as lower
alkoxy, lower alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy,
aryl, aryloxy, heterocyclic group and heterocyclic-oxy group at the
terminal of the .omega.-chain of which is shorter, the same as or
longer than that of normal prostanoic acid.
[0022] In preferred embodiments, the human patients are of the
Caucasian race.
[0023] In preferred embodiments, the periodic administration is at
least once a day for at least six months.
[0024] In another preferred embodiment of the invention, the
compound administered is a docosanoid.
[0025] In yet another preferred embodiment of the invention, the
compound administered is Unoprostone isopropyl.
[0026] In still another preferred embodiment of the invention, the
compound administered is 15-keto-latanoprost.
[0027] In another preferred embodiment of the invention, the
compound administered is 17-phenyl-18, 19, 20-trinor
PGF.sub.2.alpha. N-ethylamide
15-keto-17-phenyl-18,19,20-trinor-PGF.sub.2.alpha. N-ethylamide or
13,14-dihydro-15-keto-17-phenyl-18,19,20-trinor PGF.sub.2.alpha.
N-ethylamide.
[0028] In still another preferred embodiment of the invention, the
compound administered is a 15-keto-16-(3-trifluoro methyl
phenoxy)-17,18,19,20-tetranor PGF.sub.2.alpha. or a 13,
14-dihydro-15-keto-16-(3-trifluoromethyl
phenoxy)-17,18,19,20-tetranor PGF.sub.2.alpha., especially their
isopropyl esters.
DETAILED DESCRIPTION OF THE INVENTION
[0029] The present invention is directed to the long-term care and
management of intraocular pressure and glaucoma in human patients.
Latanoprost, 13,14-dihydro-17-phenyl-18,19, 20-trinor
PGF.sub.2.alpha. isopropyl ester, has been used in such treatment,
but causes brown pigmentation of the iris of Caucasians in a
significant number of patients (10% or more). Since it is not
believed at present possible to predict with relative certainty in
which patient iridic pigmentation will occur, this is a significant
side effect in a color-conscious world, especially for female
patients. The iridic pigmentation usually occurs by three or more
months with continuous treatment, i.e., periodic administration on
a daily basis. It is believed that the iridic pigmentation results
from latanoprost's high specific binding affinity for the
prostaglandin FP or EP receptor. (Journal of Japan Glaucoma
Society, 5, 136 (1995))
[0030] Now, it has been found that a prostaglandin related compound
of the present invention which substantially does not stimulate the
prostaglandin FP receptor or possesses an FP specific affinity
one-tenth or less than that of latanoprost and is otherwise usable
as a topically administered ocular hypotensive, can be safely
administered to humans over prolonged time periods without causing
dark colored iridic pigmentation. Certain of these compounds are
those in which carbon atom number 15 is substituted by an oxo group
(15-keto compounds), or those in which carbon atom number 15 is
substituted by a hydroxy group and the omega chain beyond carbon
atom number 15 contains a straight chain of at least 6 carbon atoms
or a straight chain of at least 3 carbon atoms with a ring at the
terminal of the omega chain. As above discussed, the compounds of
this invention can safely be administered topically for ocular
hypotensive effect to human patients over prolonged time periods
without causing the brown iridic pigmentation found with
Latanoprost.
[0031] Prostaglandin related compounds of the present invention and
of the following formula (I) are one preferred embodiment. 2
[0032] wherein W.sub.1, W.sub.2 and W.sub.3 are carbon or oxygen
atoms,
[0033] L, M and N are hydrogen atom, hydroxy, halogen atom, lower
alkyl, lower alkoxy, hydroxy(lower)alkyl, or oxo, wherein at least
one of L and M is a group other than hydrogen, and the
five-membered ring may have at least one double bond;
[0034] A is --CH.sub.2OH, --COCH.sub.2OH, --COOH or a functional
derivative thereof;
[0035] B is single bond, --CH.sub.2--, --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--;
[0036] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon residue, which is unsubstituted or
substituted with halogen, an alkyl group, hydroxy, oxo, aryl or
heterocyclic group; and
[0037] Ra is a saturated or unsaturated lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen atom, oxo, hydroxy, lower alkyl, lower alkoxy, lower
alkanoyloxy, cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl,
aryloxy, heterocyclic group or heterocyclic-oxy group;
cyclo(lower)alkyl; cyclo(lower)alkyloxy; aryl; aryloxy;
heterocyclic group; heterocyclic-oxy group.
[0038] A group of particularly preferable compounds among the above
described compounds is represented by the general formula (II):
3
[0039] wherein L, M, R.sub.1, A and B are the same definitions
described above.
[0040] X.sub.1 and X.sub.2 are hydrogen, lower alkyl, or
halogen;
[0041] R.sub.2 is a single bond or lower alkylene; and
[0042] R.sub.3 is lower alkyl, lower alkoxy, cyclo(lower)alkyl,
cyclo(lower)alkyloxy, aryl, aryloxy, heterocyclic group or
heterocyclic-oxy group.
[0043] Another preferred embodiment of this invention resides in
prostaglandin related compounds of the present invention and of the
formula (III): 4
[0044] wherein W.sub.1, W.sub.2 and W.sub.3 are carbon or oxygen
atoms,
[0045] L, M and N are hydrogen atom, hydroxy, halogen atom, lower
alkyl, lower alkoxy, hydroxy(lower)alkyl, or oxo, wherein at least
one of L and M is a group other than hydrogen, and the
five-membered ring may have at least one double bond;
[0046] A is --CH.sub.2OH, --COCH.sub.2OH, --COOH or a functional
derivative thereof;
[0047] B is single bond, --CH.sub.2--, --CH.sub.2--CH.sub.2--,
--CH.ident.CH--, --C--C--, --CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.ident.CH--,
--C.dbd.--C--CH.sub.2--, or --CH.sub.2-- C.ident.C--;
[0048] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon residue, which is unsubstituted or
substituted with halogen, an alkyl group, hydroxy, oxo, aryl or
heterocyclic group;
[0049] Z is 5
[0050] wherein R.sub.4 and R.sub.5 are hydrogen atom, hydroxy,
halogen atom, lower alkyl, lower alkoxy or hydroxy(lower)alkyl,
wherein R.sub.4 and R.sub.5 are not hydroxy, lower alkoxy and/or
hydroxy (lower) alkyl at the same time; and
[0051] Ra' comprises (1) a saturated or unsaturated C.sub.3 to
C.sub.5 straight chain aliphatic hydrocarbon beyond carbon atom
number 15 substituted by cyclo (lower) alkyl, cyclo (lower)
alkyloxy, aryl, aryloxy, heterocyclic group or heterocyclic oxy
group at its terminus or (2) a saturated or unsaturated at least
C.sub.6 straight chain aliphatic hydrocarbon residue beyond carbon
atom number 15, which is unsubstituted or substituted with halogen
atom, oxo, hydroxy, lower alkyl, lower alkoxy, lower alkanoyloxy,
cyclo(lower)alkyl, cyclo(lower)alkyloxy, aryl, aryloxy,
heterocyclic group or heterocyclic-oxy group.
[0052] Still another preferred embodiment of this invention resides
in prostaglandin related compounds of the present invention and of
the formula (IV): 6
[0053] wherein W.sub.1, W.sub.2, and W.sub.3 are carbon or oxygen
atoms,
[0054] L, M and N are hydrogen atom, hydroxy, halogen atom, lower
alkyl, lower alkoxy, hydroxy(lower)alkyl, or oxo, wherein at least
one of L and M is a group other than hydrogen, and the
five-membered ring may have at least one double bond;
[0055] R' and R" is hydrogen atom, lower alkyl, aryl, alkyl- or
aryl-sulfonyl, lower alkenyl and lower alkynyl;
[0056] B is single bond, --CH.sub.2--, --CH.sub.2--CH.sub.2--,
--CH.dbd.CH--, --C.dbd.C--, CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--, --CH.sub.2CH.dbd.CH--,
--C.ident.C--CH.sub.2--, or CH.sub.2--C.ident.C--
[0057] R.sub.1 is a saturated or unsaturated bivalent lower or
medium aliphatic hydrocarbon residue, which is unsubstituted or
substituted with halogen, an alkyl group, hydroxy, oxo, aryl or
heterocyclic group;
[0058] Z is 7
[0059] wherein R.sub.4 and R.sub.5 are hydrogen atom, hydroxy,
halogen atom, lower alkyl, lower alkoxy or hydroxy(lower)alkyl,
wherein R.sub.4 and R.sub.5 are not hydroxy, lower alkoxy and/or
hydroxy(lower)alkyl at the same time; and
[0060] Ra is a saturated or unsaturated lower or medium aliphatic
hydrocarbon residue, which is unsubstituted or substituted with
halogen atom, oxo, hydroxy, lower alkyl, lower alkoxy, lower
alkanoyloxy, cyclo(lower)alkyl, cyclo(lower))alkyloxy, aryl,
aryloxy, heterocyclic group or hetrocyclic-oxy group;
cyclo(lower)alkyl; cyclo(lower)alkyloxy; aryl; aryloxy;
heterocyclic group; heterocyclic-oxy group.
[0061] The compounds used with the present invention can be of the
prostaglandins A, B, C, D, E, F, or J type and include subtypes 1,
2, and 3, all as explained in U.S. Pat. No. 5,001,153, the entire
content of which is incorporated herein by reference. Compounds
usable in the present invention are described in U.S. Pat. No.
5,001,153, and in U.S. Pat. No. 5,312,128, including their
ophthalmic preparations.
[0062] In the above formula, the term "unsaturated" in the
definitions for R.sub.1, Ra 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.
[0063] 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 1 to 8 carbon atoms for R.sub.1 and
1 to 10, especially 1 to 8 carbon atoms for Ra.
[0064] The term "halogen atom" covers fluorine, chlorine, bromine
and iodine. Particularly preferable is a fluorine atom.
[0065] The term "lower" throughout the specification is intended to
include a group having 1 to 6 carbon atoms unless otherwise
specified.
[0066] 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.
[0067] The term "lower alkoxy" refers to a group of lower
alkyl--O--, wherein lower alkyl is as defined above.
[0068] 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- .
[0069] 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.
[0070] 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.
[0071] The term "cyclo(lower)alkyloxy" refers to the group of
cyclo(lower)alkyl--O--, wherein cyclo(lower)alkyl is as defined
above.
[0072] The term "aryl" may include unsubstituted or substituted
aromatic hydrocarbon rings (preferably monocyclic groups), for
example, phenyl, naphthyl, tolyl, and xylyl. Examples of the
substituents are halogen atom, lower alkoxy and halo(lower)alkyl,
wherein halogen atom and lower alkyl are as defined above.
[0073] The term "aryloxy" refers to a group represented by the
formula ArO--, wherein Ar is aryl as defined above.
[0074] The term "heterocyclic group" may include mono- to
tri-cyclic, preferably monocyclic heterocyclic group which has a 5
to 14, preferably 5 to 10 membered ring having optionally
substituted carbon atom(s) and 1 to 4, preferably 1 to 3, of 1 or 2
types 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, pyridazyl,
pyrimidyl, pyrazyl, 2-pyrrolinyl, pyrrolidinyl, 2-imidazolinyl,
imidazolidinyl, 2-pyrazolinyl, pyrazolidinyl, piperazinyl,
morpholino, indolyl, benzothienyl, quinolyl, isoquinolyl, puryl,
quinazolinyl, carbazolyl, acridinyl, phenanthridinyl,
benzimidazolyl, benzimidazolonyl, 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.
[0075] The term "heterocyclic-oxy group" means a group represented
by the formula HcO--, wherein Hc is a heterocyclic group as
described above.
[0076] The term "functional derivative," of A includes salts
(preferably pharmaceutically acceptable salts), ethers, esters and
amides.
[0077] 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.
[0078] 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.
[0079] 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, tosyl
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.
[0080] The amides 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.
[0081] Preferred examples of L and M include hydroxy and oxo, and
especially, M and L are hydroxy to provide a 5-membered ring
structure of, so called, PGF type.
[0082] Preferred A is --COOH, --CH.sub.2OH, or its pharmaceutically
acceptable salt, ester, ether or amide thereof.
[0083] Preferred example of X.sub.1 and X.sub.2 is 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.
[0084] Preferred R.sub.1 is an unsubstituted saturated or
unsaturated bivalent lower-medium aliphatic hydrocarbon residue. It
may preferably have 1-10 carbon atoms, more preferably, 2-8 carbon
atoms.
[0085] Examples of R.sub.1 include, for example, the following
groups:
[0086] --CH.sub.2--CH.sub.2--,
[0087] --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
[0088] --CH.sub.2--CH.dbd.CH--CH.sub.2--,
[0089] --CH.sub.2--C.ident.--C--CH.sub.2--,
[0090] --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
[0091] --CH.sub.2--CH.dbd.CH--CH.sub.2--CH.sub.2--,
[0092] --CH.sub.2--C.ident.--C --CH.sub.2--CH.sub.2--,
[0093]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--,
[0094] --CH.sub.2--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--,
[0095] --CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.dbd.CH--,
[0096] --CH.sub.2--C.ident.C --CH.sub.2--CH.sub.2--CH.sub.2--,
[0097] --CH.sub.2--CH.sub.2--CH.sub.2 --H.sub.2 --CH (CH.sub.3)
--CH.sub.2--
[0098]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.su-
b.2--CH.sub.2--,
[0099] --CH.sub.2--CH.ident.CH --CH.sub.2 --CH.sub.2 --H.sub.2
--CH.sub.2 --CH.sub.2--,
[0100]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.db-
d.CH--,
[0101]
--CH.sub.2--C.dbd.--C--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.s-
ub.2--,
[0102]
--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.sub.2--CH.su-
b.2(CH.sub.3)--H.sub.2--,
[0103] Preferred R.sub.2 is a single bond or a saturated or
unsaturated bivalent lower to medium aliphatic hydrocarbon residue,
which may preferably have 1-10 carbon atoms, more preferably 1-8
carbon atoms, especially 1-6 alkylene.
[0104] Preferred R.sub.3 is a hydrogen atom, aryl or aryloxy.
[0105] Preferred Ra is a hydrocarbon containing 1-10 carbon atoms,
more preferably, 1-8 carbon atoms and, especially, that having 7
carbon atoms beyond carbon atom number 15.
[0106] Preferred Ra' is a straight chain beyond carbon atom number
15 of at least 6 carbon atoms, or at least 3 carbon atoms with a
ring, more preferably a phenyl ring, at the terminal of the omega
chain.
[0107] Preferred R' and R" is hydrogen atom or C1-6 alkyl,
C.sub.2-6 alkenyl and C.sub.3-6 alkynyl.
[0108] The configuration of the ring and the .alpha.- and/or
.omega. chains in the present invention 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.
[0109] When a 15-keto-PG compound of the present invention has for
example a single bond between carbon atom number 13 and 14, the
compound may be in the keto-hemiacetal equilibrium by formation of
a hemiacetal between hydroxy at position 11 and oxo at position
15.
[0110] If such tautomeric isomers as above are present, the
proportion of both tautomeric isomers varies with the structure of
the rest of the molecule or the kind of the substituent present.
Sometimes one isomer may predominantly be present in comparison
with the other. However, it is to be appreciated that the compounds
used in the invention include both isomers. Further, while the
compounds used in the invention may be represented by a structure
formula or name based on keto-type regardless of the presence or
absence of the isomers, it is to be noted that such structure or
name does not intend to exclude the hemiacetal type compound.
[0111] The present invention includes any of the isomers such as
the individual tautomeric isomers, a mixture thereof, or the
optical isomers, a mixture thereof, a racemic mixture, and other
steric isomers useful for the same purpose.
[0112] Compounds useful in the practice of this invention are
13,14-dihydro-15-keto-20-ethyl PGF.sub.2.alpha. isopropyl ester;
13,14-dihydro-15-keto-17-phenyl-18,19,20-trinor-PGF.sub.2.alpha.
isopropyl ester;
17-phenyl-18,19,20-trinor-PGF.sub.2.alpha.N-ethylamide;
15-keto-17-phenyl-18,19,20-trinor PGF.sub.2.alpha. N-ethylamide;
13,14-dihydro-15-keto-17-phenyl-18,19,20-trinor PGF.sub.2.alpha.
N-ethylamide; 15-keto-16-(3-trifluoromethyl
phenoxy)-17,18,19,20-tetranor PGF.sub.2.alpha. isopropyl ester and
13,14-dihydro-15-keto-(3-trifluorome- thyl
phenoxy)-17,18,19,20-tetranor PGF.sub.2.alpha. isopropyl ester.
[0113] In the method of the present invention, the above-described
compounds are topically administered to the affected eye once or
twice a day for at least six months, up to a time period as long as
required, to complete treatment for elevated ocular pressure or
glaucoma, as needed, or to maintain a prophylactic level for six
months or longer, without causing iridic pigmentation, especially
the brown pigmentation caused by long term use of Latanoprost.
[0114] The pharmaceutical composition of the present invention
include ophthalmic solution and ointment. The ophthalmic solution
may be prepared by dissolving the active ingredient into sterilized
aqueous solution such as saline or buffer. A powder composition for
ophthalmic solution to be dissolved before use may also be used.
The ophthalmic ointment may be prepared by mixing the active
ingredient with ointment base.
[0115] The pharmaceutical composition of the present invention will
usually have a concentration of less than approximately 0.20% (w/v)
of active compound. Preferred concentrations are usually within the
concentration range of about 0.00005 to 0.18% (w/v).
[0116] The composition of the present invention may further be
admixed with any of pharmaceutically active agents in so far as
said agent is compatible with the purpose of the present
invention.
EXAMPLE 1
[0117] 813 human subjects, mostly Caucasian, were studied for about
one year. One drop of 0.15% Rescula.RTM. was administered to each
treated eye twice a day. After about the first six months of the
trial, there was no reported increase in brown pigmentation of the
iris.
EXAMPLE 2
IN VITRO FUNCTIONAL RECEPTOR STUDIES
FP-receptor activity
[0118] Functional FP-receptor studies were performed using iris
sphincter muscles from cat eyes. The eyes were either used directly
after enucleation or stored in ice cold saline overnight. The iris
sphincter muscles were prepared, cut in halfs, and mounted in
thermostated (37.degree. C.) tissue baths with oxygenated modified
Kreb's solution containing indomethacin (2.8.times.10.sup.-6 M),
atropine (10.sup.-7 M) and propranolol (10.sup.-7 M). A resting
tension of 150 mg was applied, and the contractile force was
measured isometrically after cumulative dosing of test compounds. A
manual or automated tissue bath system (Buxco, STC 400) was used
for the experiments. For each tissue sample the maximal response
was normalized to 100%. The mean response of two to four different
preparations was calculated.
EP.sub.1-receptor activity
[0119] Iris sphincter muscles from bovine eyes were used for
studies of EP.sub.1-receptor responses. Freshly enucleated bovine
eyes were transported on ice from a slaughterhouse. The experiments
were performed essentially in the same way as with cat iris
sphincters, but since bovine iris sphincter expresses both EP.sub.1
and TP-receptors, a TP-receptor agonist (GR32 191B) was added to
the tissue baths in concentration of 0.1 .mu.M.
EP.sub.2-receptor activity
[0120] Segments of guinea-pig ileum were used to study the effect
on EP.sub.2-receptors. Male guinea pigs were killed with a blow to
the head and bled from the carotid arteries. Circular rings (2-3 mm
wide) were taken from the ileum about 10 cm from the caecum, a
resting tension of 1 g was applied and the preparation was
stimulated by using a Gras S88 stimulator delivering trains of
square pulses (frequency 7.5 Hz, pulse duration 0.1 msec, train
duration 4 sec) to two parallel platinum electrodes in the bath.
The voltage over the electrodes was approximately 20 V, and the
interval between trains was 100 seconds. The peak contractile force
at each stimulation train was measured. Test compounds were added
cumulatively, and the inhibitory effect on the muscle twitch was
evaluated. A response was taken when three successive peaks had a
similar height.
EP.sub.3-receptor activity
[0121] Guinea-pig vas deferens was used to assess the effect on
EP.sub.3-receptors. Tissue segments of about 10 mm were mounted in
tissue baths and the experiments were performed essentially in the
same way as described for the guinea-pig circular ileum.
Platelet studies of TP and IP/DP receptor activity
[0122] The activity of test compounds on platelet aggregation was
performed on samples from guinea-pig blood. Blood was collected in
Venoject tubes containing sodium citrate (to give a concentration
of 10 mM) and centrifuged at 200 G for 10 minutes to obtain
platelet rich plasma (PRP). The PRP was removed and the remainder
was further centrifuged 2000 G for 15 minutes and the supernatant
was used as platelet poor plasma (PPP). Platelet aggregation was
measured in a Chrono-Log aggregometer using PPP as a blank. 500
.mu.l PRP was incubated for 2 minutes at 37.degree. C. and 5 .mu.l
of the test solution was added. The maximal change in light
transmission was recorded.
[0123] Aggregation of guinea-pig thrombocytes was used to measure
the effect on TP receptors. The maximal change in light
transmission for each test solution was calculated in percent of
the maximal aggregation obtained with stable TxA2 analogue U46619
(3 .mu.M) in the same batch of PRP.
[0124] The activity on IP and DP-receptors was determined from the
ability of the compounds to inhibit ADP induced aggregation of
guinea pig PRP. The test substance (5 .mu.l) was added to 500 .mu.l
PRP and incubated at 37.degree. C. for two minutes. ADP (final
concentration 10 .mu.M) was added and the maximal change in light
transmission induced by ADP in absence and presence of the test
drug in the same batch of PRP was measured and expressed in
percent.
[0125] Concentration-effect curves were fitted to the data using
the logistic sigmoid function.
[0126] Table I.
[0127] Prostaglandin receptor profile of latanoprost acid and
15-keto-latanoprost acid based on EC.sub.50 values (moles/l)
obtained in smooth muscle bath experiments
1 PG-analogues FP EP1 EP2 EP3 Latanoprost 1.0 .times. 10.sup.-8 4.9
.times. 10 5.3 .times. 10.sup.-4 2.8 .times. 10.sup.-5 Acid I5-keto
Latanoprost 2.6 .times. 10.sup.-7 2.8 .times. 10.sup.-5
>10.sup.-4 >10.sup.-4 Acid
[0128] The results in Table I indicate that a concentration of
latanoprost acid of only 1.0.times.10.sup.-8 M is needed to
stimulate the FP-receptor at 50% efficacy compared to significantly
higher concentrations for the other prostaglandin receptors. This
means that latanoprost is a selective agonist for the FP receptor
and exerts its biological effect primarily through stimulation of
this receptor when administered to the eye in clinical doses. A
considerably higher concentration of 2.6.times.10.sup.-7 M was
required for 15-keto-Iatanoprost acid for a similar stimulation of
the FP-receptor. Accordingly, the receptor profile study of Table I
provides evidence that 15-keto-latanoprost acid is at least ten
times less potent than latanoprost acid with respect to the
activity on the FP receptor, and has virtually no activity on the
other prostaglandin receptors (EP.sub.1 EP.sub.2, and
EP.sub.3.).
EXAMPLE 3
AFFINITY PROFILE OF UNOPROSTONE FOR PROSTAGLANDIN RECEPTORS
[0129] The studies undertaken were to determine whether Unoprostone
isopropyl or its metabolite (the acid obtained by hydrolysis of the
isopropyl ester) has affinity for any prostaglandin receptor.
[0130] Ligand binding assays and signal transduction studies.
[0131] In ligand binding studies, the specificity of labeled
Unoprostone isopropyl and the metabolite binding to prostaglandin
receptor sites in bovine corpus luteal membranes was determined.
This tissue expresses most of the PG receptors including an
abundance of FP receptors. In signal transduction studies, the
mobilization of intracellular calcium using Fluorescence Imaging
technique in the primary culture of human ciliary muscle cells and
cyclic AMP generation in rabbit iris-ciliary body was measured.
METHODS
[0132] 1) Ligand binding assay method
[0133] This method is well described in the literature. Briefly,
8-16 nM .sup.3H-PGF.sub.2.alpha. or .sup.3H- Unoprostone isopropyl
or the metabolite was incubated in the presence or absence of
1,000-fold excess of unlabeled PGF.sub.2.alpha. or Unoprostone
isopropyl or its metabolite with bovine corpus luteal membranes.
Free unbound ligands were separated by rapid filtration using a
millipore filtration assembly. Membrane bound radioactivity
retained by the filter was counted. It is important to define the
following terms for appreciation of the binding data:
[0134] Total binding is defined as the amount bound in the absence
of unlabeled ligand.
[0135] Specific binding is defined as the binding displaced by
unlabeled ligand.
[0136] Non-specific binding is the binding not displaced by
unlabeled ligand.
[0137] 2) Signal transduction methods
[0138] Intracellular calcium mobilization
[0139] Human ciliary muscle cells at confluency were loaded with
Fura 2-AM. Cells in a petri dish were put in a jacketed chamber
mounted on the microscope platform. The chamber was maintained at
37.degree. C. by thermostatically controlled water circulating
through the chamber jacket. The cells were then exposed to the
excitation wavelengths of 334 and 380 nm with an emission
wavelength of 510 nm. Cells were perfused with HEPES buffer with or
without test compounds. The intracellular calcium was visualized
and photographed using Attofluor RatioVision software program and
Zeiss fluorescence microscope.
Results
[0140] 1) Binding Studies
[0141] To optimize the specific binding of .sup.3H-Unoprostone
isopropyl or the metabolite, initially binding was determined as
the function of time, membrane and radioligand concentrations in
the presence or absence of unlabeled Unoprostone or
PGF.sub.2.alpha.. At 5 minutes, total binding was 21.0 fmoles/mg
protein, which did not increase with time up to 60 minutes in
bovine corpus luteal membranes.
[0142] Tables II and III showed that total binding of
.sup.3H-Unoprostone isopropyl increased with concentrations of the
membrane and radioligand but none of the binding was displaced by
unlabeled Unoprostone isopropyl or PGF.sub.2.alpha. suggesting no
specific binding.
[0143] Table II.
[0144] Total Binding of .sup.3H-Unoprostone isopropyl in bovine
corpus luteal membranes as a function of membrane
concentration.
2 Total binding: fmoles/mg Protein Conc. protein 50 .mu.g 13.0 .+-.
2.0 (3) 100 .mu.g 23.0 .+-. 6.0 (3) 200 .mu.g 32.0 .+-. 8.0 (3)
[0145] Unlabeled Unoprostone isopropyl or PGF.sub.2.alpha. did not
displace any of the above binding.
[0146] Table III.
[0147] Binding of .sup.3H-Unoprostone isopropyl and
.sup.3H-PGF.sub.2.alpha. in bovine corpus luteal membranes as a
function of concentration.
3 .sup.3H-Unoprostone isopropyl .sup.3H-PGF2.alpha. Total Specific
Specific Binding Binding Binding Binding Conc. fmoles/mg fmoles/mg
Conc. fmoles/mg fmoles/mg (nM) protein protein (nM) protein protein
8.0 19.4 .+-. 2.0(3) 0 2.0 150.0 .+-. 16.0(3) 120.0 .+-. 15.0(3)
16.0 25.5 .+-. 4.0(3) 0 4.0 325.0 .+-. 25.0(3) 261.0 .+-. 27.0(3)
32.0 40.0 .+-. 7.0(3) 0 8.0 505.0 .+-. 75.0(3) 423.0 .+-.
40.0(3)
[0148] There were no specific binding sites for Unoprostone
isopropyl as shown in the Table IV because none of the unlabeled
Unoprostone isopropyl concentrations displaced the bound
.sup.3H-Unoprostone isopropyl. In contrast,
.sup.3H-PGF.sub.2.alpha. was bound specifically to its binding
sites. Competition studies showed that 17-phenyl trinor PGE.sub.1
(EP.sub.1), Butaprost (EP.sub.2), sulprostone (EP.sub.3) , iloprost
(IP), U46619 (TP) and BW245C (DP) did not displace
.sup.3H-Unoprostone isopropyl from the binding sites suggesting
that Unoprostone isopropyl does not bind to DP, EP.sub.1, EP.sub.2,
EP.sub.3, IP and TP receptor sites. The metabolite has similar
binding profiles as Unoprostone isopropyl, i.e. it does not bind to
any PG receptor sites.
[0149] Table IV.
[0150] Competition for .sup.3H-Unoprostone isopropyl or
PGF.sub.2.alpha. binding sites by prostaglandin receptor
agonists
4 Total .sup.3H-Uno- Specific Total Competing prostone Binding Com-
.sup.3H-PGF.sub.2.alpha. Ligand Uno- isopropyl of .sup.3H- peting
binding Specific prostone binding Uno- Ligand nmoles/ Binding
Isopropyl fmoles/mg prostone PGF.sub.2.alpha. mg fmoles/mg (.mu.M)
protein isopropyl (.mu.M) Protein Protein 2.5 7.0 0 0.062 0.56 0.07
5.0 6.5 0 0.25 0.56 0.15 8.0 6.7 0 0.5 0.56 0.25 16.0 7.6 0 8.0
0.56 0.42
[0151] 2) Second messenger study: Calcium mobilization
[0152] Unoprostone isopropyl or the metabolite at concentrations
ranging from 10-1000 nM did not mobilize intracellular calcium in
human ciliary muscle cells. Fluprostenol, a FP receptor agonist
linked to intracellular calcium mobilization pathway, mobilized
calcium. The inability of Unoprostone isopropyl or the metabolite
to mobilize intracellular calcium suggests that these compounds
have no affinity for EP.sub.1 or FP or TP receptors. These
receptors are linked to phospholipase C pathway, stimulation of
which results in the mobilization of intracellular calcium.
Conclusions
[0153] The results of the studies demonstrated that Unoprostone
isopropyl or the metabolite does not bind to PG receptor sites.
[0154] In addition to the above Examples 2 and 3, Goth et al,
Japanese Journal Ophthalmol, 36: 236-245 (1994), investigated the
agonistic activity and affinity of Unoprostone (the free acid form)
and PhXA34 (also in the free acid form). PhXA34 is an epimeric
mixture of 15R and 15S compounds thereby including latanoprost and
its 15S-epimer. PhXA34 exhibited 1/2 the agonistic activity of
PGF.sub.2.alpha., to PGF.sub.2.alpha., receptors, as determined
from contraction of cat iris sphincter, while Unoprostone's
agonistic activity was only {fraction (1/1600)} of that of
PGF.sub.2.alpha. Binding inhibition studies showed that PhXA34
strongly inhibited [.sup.3H] PGF.sub.2.alpha. binding using bovine
corpus luteum, which has PGF.sub.2.alpha. receptors while the
binding inhibitory activity of Unoprostone was infinitely weaker
than that of PGF.sub.2.alpha. (<<{fraction (1/280)} of
PGF.sub.2.alpha.
[0155] The terminology "causing less pigmentation than latanoprost"
means over the same treatment time period of at least six months
using a clinically approved dosage of once or twice per day, that
the compound used in accordance with the present invention causes
less irridic pigmentation in comparable irides and/or causes a
reduced frequency of pigmentation in a patient population.
[0156] The compounds usable in the present invention are at least
five times less potent than latanoprost in stimulating the FP
receptor as determined by the test of Example 2 herein. Preferably,
the usable compounds are at least about ten times less potent than
latanoprost regarding stimulation of the FP receptor. Most
preferably, the usable compounds also are those exhibiting
virtually no stimulation activity on the EP.sub.1, EP.sub.2,
EP.sub.3, TP and IP/DP receptors.
[0157] Variations of the invention will be apparent to the skilled
artisan.
INDUSTRIAL APPLICABILITY
[0158] The process, composition and use of the present invention
are useful for long-term treatment of ocular hypertension and
glaucoma without causing pigmentation or with causing comparatively
minimal pigmentation of the iris.
* * * * *