Note: Descriptions are shown in the official language in which they were submitted.
2138181
USE OF CERTAIN pRo8T~aT~NnIN ~N~TO5UE8
TO TREAT GLA~COMA AND OCULAR HYPERTEN8ION
Background of the Invention
The present invention relates to the use of
certain prostaglandins and prostaglandin analogues for
the treatment of glaucoma and ocular hypertension. As
used herein, the terms "prostaglandin" and "PG" shall
refer to prostaglandins and derivatives and analogues
thereof, except as otherwise indicated by context.
Naturally-occurring prostaglandins are known to
lower intraocular pressure (IOP) after topical ocular
instillation, but generally cause inflammation, as well
as surface irritation characterized by conjunctival
hyperemia and edema. Many synthetic prostaglandins
have been observed to lower intraocular pressure, but
such compounds also produce the aforementioned side
effects. Various attempts have been made to overcome
the ocular side effects associated with prostaglandins.
Stjernschantz et al. (EP 364 417 B1) have synthesized
derivatives or analogues of naturally-occurring
prostaglandins in order to design out selectively the
undesired side effects while maintaining the IOP-
lowering effect. Others, including Ueno et al. (EP 330
511 A2) and Wheeler (EP 435 682 A2) have tried
complexing prostaglandins with various cyclodextrins.
Summary of the Invention
It has now been unexpectedly discovered that
certain D series prostaglandin analogues are
significantly more effective in lowering IOP than other
known prostaglandins. In particular, the prostaglandin
analogues of the present invention have unexpectedly
2138181
been found to lower IOP to a greater degree than other
known PGs without significant inflammation or
associated side effects which often accompany topical
ocular administration of D series prostaglandins.
Detailed DescriPtion of the Invention
The prostaglandins which are useful in the
compositions of the present invention have the general
formulae (I) and (II): .
X~,~"~R,
R, (I)
wherein:
Rl = CO2Rz, wherein R2 = H, a cationic salt moiety,
or an ophthalmically acceptable ammonium
moiety; or Rl may also represent an
ophthalmically acceptable ester moiety;
X = halogen, particularly Cl or F, in either
configuration;
Y = CH2 or O;
--- = single or trans double bond;
R3, R4 can be the same or different, and are
selected from: free or functionally modified
hydroxy groups; and
n = 0 or 1.
X R~ R2
<~""`~A~W X
~Y~
~ (II)
wherein:
Rl = CH2R, CO2R4;
21381~1
R = OH or functionally modified hydroxy group;
R2 and R3 can be the same or different and are
selected from: H and CH3;
R4 = H, a cationic salt moiety, an ophthalmically
acceptable ammonium moiety, substituted or
unsubstituted alkyl, cycloalkyl,
(cycloalkyl)alkyl, aryl, arylalkyl,
heteroaryl, or (heteroaryl)alkyl, wherein
substituents include alkyl, halo, a free or
functionally modified hydroxy group, or a
free or functionally modified thiol;
W = CH2, O, S()m wherein m = O, 1, 2;
A = CH2CH2, cis or trans CH=CH, or CQC;
X = Cl, F or R in either configuration, or H;
Zl1 may be selected from O (i.e. a carbonyl), H and
R in either configuration, or H and H;
Z,5 may be selected from O (i.e., a carbonyl) H and
R in either configuration or H and H;
Y = CH2CH2 or trans CH=CH, or CQC; and
n = 0 or 1.
As used in this specification, the term
"ophthalmically acceptable ester moiety" refers to an
ester moiety which is non-toxic and does not cause
undue irritation to the ocular tissues. For compounds
of formula (I), examples of ophthalmically acceptable
esters include, but are not limited to: R2 =
substituted or unsubstituted alkyl, cycloalkyl,
(cycloalkyl)alkyl, aryl, arylalkyl, heteroaryl, or
(heteroaryl)alkyl, wherein substituents include alkyl,
halo, a free or functionally modified hydroxy group, or
a free or functionally modified thiol. As used in this
specification, the term "heteroaryl" refers to a
monocyclic ring system of 5 or 6 atoms composed of C,
N, O, and/or S, such as furan, thiophene, pyrrole,
pyrazole, imidazole, triazole, tetrazole, oxazole,
isothiazole, thiazole, thiadiazole, pyridine,
21381~1
pyrimidine, pyradazine and pyrazine. The term
"functionally modified hydroxy groups" refers to either
etherified hydroxy groups or acylated (esterified)
hydroxy groups. Similarly, the term "functionally
modified thiol" refers to an alkylated thiol.
Preferred compounds of formula (I) include those
wherein: Rl = CO2R2; R2 = H, methyl, ethyl, n-propyl,
isopropyl, t-butyl, or benzyl; X = Cl in the ~ (R)
configuration; Y = O or CH2; R3 and R4 = OH; and n = 1.
The most preferred compounds of formula (I) are those
wherein: Rl = CO2R2; R2 = H, methyl, ethyl, isopropyl,
or t-butyl; X = Cl in the ~ (R) configuration; Y = O;
R3 and R4 = OH; and n = 1.
Preferred compounds of formula (II) include those
wherein: W = CH2 or 0; A = cis CH=CH; X = Cl or H; Z
= R and H in either configuration or H and H; Y =
CH2CH2, or trans CH=CH; and n = 1. The most preferred
compounds of formula (II) are those wherein: R2, R3 =
H; X = Cl in ~ (Rj configuration or H; Zll = H and R in
either configuration; Z15 = H and R in either
configuration; R1=CO2R4; and R4=H, a cationic salt
moiety, or substituted or unsubstituted C1-C10 alkyl.
Some of the compounds of formula (II) are novel.
The novel compounds of formula (II) are those wherein:
25 Rl = CH2R, CO2R4; R = OH or a functionally modified
hydroxy group; R2 and R3 can be the same or different
and are selected from: H and CH3; R4 = H, a cationic
salt moiety, substituted or unsubstituted alkyl,
cycloalkyl, (cycloalkyl)alkyl, aryl, arylalkyl,
heteroaryl, or (heteroaryl)alkyl, wherein substituents
include alkyl, halo, a free or functionally modified
hydroxy group, or a free or functionally modified
thiol; W = O, S(O)m, wherein m = O, 1, 2; A = CH2CH2,
cis or trans CH=CH, or CQC; X = H; Zll and Z15 may be the
2138181
same or different and may be selected from O (i.e., a
carbonyl), H and R in either configuration, or H and
H; Y = CH2CH2 or trans CH=CH, or CÇC; and n = O or 1.
The preferred novel compounds of formula (II) are
those wherein: Rl = CO2R4; R2 and R3 = H; R4=H, a cationic
salt moiety, or a substituted or unsubstituted Cl-C10
alkyl; W = O, A = cis CH=CH; Zll = R and H in either
configuration; Y = CH2CH2, or trans CH=CH; Z15 = R and H
in either configuration; and n = O or 1.
The following Table 1 contains examples of some
preferred compounds of the present invention.
213Sl~l
- TAR~
COMPOUND NAME COMPOUND STRUCTURE
III (5Z, 13E)~9R,llR,lSS)-9 Chloro-15- cl ~
cyclohexyl-11,15 dihydroxy~ ` CO2CH3
16,17,18,19,2~1t~or-5,13- ~ I ~
prostadienoic Acid Methyl Ester à~ ~¦
OH
IV (5Z, 13E)~9R,llR 155)-9 Chloro-15- a ~
cyclohexyl-11,15 dihyd~oA~-3-oxa- ~ . O co2c(cH3)s
16,17,18,19,20-~1~anor-5,13- < I
prostadienoic Acid t-Butyl Ester
OH
V (5Z)~9R, 11R, 15R)-9~hloro 15- a ` ~
cyclohexyl-11,15 dihydroxy-3-oxa- ~ . ` ~o co2C~CH3)3
16,17,18,19,2~nl~lor-5 -~
~ros~oic Acid t-Butyl Ester
OH --
OH
Vl (5Z)~9R, 11R, 15R)-9 Chloro-15- a ``
cyclohexyl-11,15-dil~lroA~oxa- ~.: ~ co2cH(aH3)2
16,17,18,19,20--~LldlWl--S < I /~
prOsl~oic Acid I~o~l Ester ~ J -
OH
VII (5Z)~9R, 11R, 15R)~loro-15- a
cyclohexyl-11,15 dihydroxy- ~ CO2CH(CH3)2
16,17,18,19,20-~lanor-5 < I , ~,
~icAcid ~opropyl E~ter ~ J
OH
VIII (5Z)~9R, 11R, 15R)-9-Chloro-15- a
cydohexyl-ll-l~ A~ 5~ ~ = Q~co2ca~
me~oxy-3-oxa-16,17,18,19,'~ < I ~
~t~or-5 ~Y)5l~WiC Acid ~-8utyl ~J
Oa~3
2138181
IX (5E)~llR, 15~)-15~ydohexyl-
11,15-dihydroxy-3-oxa~ ~`O~CO2CH(CH3)2
16,17,18,19,20-~n~or-5-
prostadienoic Acid Isopropyl Ester ~y
HO HO
X (5E, 13E)-(llR, 15S)-15~ydohexyl-
11,15~ihydroxy-3-oxa- ~` ~O~CO2CH(CH3)2
16,17,18,19,20-~ anor-5,13- \~
prostadienoic Acid Iso~.o~l Ester HO H~--)
XI (5Z)~9R, 11R, 15R)-9~hloro-15- a
cydohexyl-11,15 dihydroxy-3-oxa- ~ .~"`~ o~CH20H
16,17,18,19,20-y~lt~u~r-5 ?ro3~nol < I I f~
~
OH
XII (5Z)~9R, 11R, 15S)-9~hloro-15- a
cydohexyl-11,15-dil.ydloA~-3-oxa- ~ ~= 0 CH20H
16,17,18,19,20-y~ll~ or-5,13
prosta~
OH
XIII (5E, 13E)-(9R, 11R, 15S)-9-Chloro- a
15-cycdohexyl 11,15 dil~y~oAr 3 ~ ~ ~ ~O~CO2C~CH~,
oxa-16,17,18,19,20-~-~or-5,13-
prostadienoic Acid t-Butyl Ester J
OH
XIV (5Z)-(9R, 11R, 15S)~loro-15- a
cydohexyl-11,15 dil~y~OAy- ~ = o~CO2CH~H~2
16,17,18,19,20-~l~.or-5-
prwtenoic Aad I~o~l Ester ~J
OH
XV (5Z)-(9R, 11R, 15R)-9-Chloro~15- a
cycdohexyl-ll,15 dil-ydroAy-3 oxa- ~ ~ Q )2a~çH~,
16,17,18,19,20-~ænta~or-5 < I~ ~
~r~5t~0ic Acit Neo~ll~rl Ester _~J
OH OH
213~
XVI (SZ,13E)-(1 lR,lSS)-lS{:yclohexyl- ~ ~"~O~CO2C~(CH3)2
ll,lS dihydroxy-3-oxa- \_1~ ~
16,17,18,19,20-~ or-5,13- ~ )
proshdienoic acid isopropyl ester HO HO ~--
Some of the above-mentioned prostaglandins are
disclosed in US 5,004,752 (Raduechel et al.) and EP 299
914 (Buchmann et al.). To the extent that US 5,004,752
and EP 299 914 teach the preparation of the
prostaglandins of the present invention, these patents
are hereby incorporated by reference herein. In
addition, the syntheses of some of the prostaglandins
of Table 1, above, are detailed below in Examples 1-12.
In the following Examples 1-12, the following
standard abbreviations are used: g = grams (mg =
milligrams); mol = moles (mmol = millimoles); mol~ =
mole percent; mL = milliliters; mm Hg = millimeters of
mercury; mp = melting point; bp = boiling point; h =
hours; and min = minutes. In addition, "NMR" refers to
nuclear magnetic resonance spectroscopy and "CI MS"
refers to chemical ionization mass spectrometry.
2138181
EXAMPLE 1: SYNTHESIS OF COMPOUND V
21381~1
A: Dimethyl (2-cyclohexyl-2-oxo)ethylPhosphonate (2):
A solution of dimethyl methylphosphonate (100 g,
0.8 mol) in 1.0 L of anhydrous THF was cooled to -70C
and n-BuLi (2.5 M in hexanes, 320 mL, 0.8 mol) was
S added dropwise such that the temperature remained below
-60C. The mixture was stirred for 10 min at -70C and
then methyl cyclohexanecarboxylate (57.3 mL, 0.4 mol)
was added dropwise, via syringe, over a period of 15
min. The resulting mixture was then stirred for 14 h
at room temperature. The reaction was quenched by
first cooling to 0C followed by the addition of 2 M
HCl until the aqueous layer was at pH 2. The layers
were separated and the aqueous layer was extracted with
2 X 200 mL of CH2Cl2. The organic layers were combined
and washed sequentially with 200 mL each of water and
brine and then dried (MgSO4). Filtration and solvent
removal gave a yellow oil which was distilled under
vacuum to afford 67.3 g (72%) of 2 as a clear colorless
liquid: bp=100-115C (0.01 mmHg); IH NMR (CDCl3) ~ 3.74
(d, J = 12.0 Hz, 6H), 3.08 (d, J = 22 Hz, 2H), 2.55 (m,
lH), l.9S - 1.60 (m, SH), 1.40 - l.lS (m, SH).
B: (3aR, 4R, SR, 6aS)-S-(Benzoyloxy)-4-[(E)-3-
cyclohexyl-3-oxo-1-propenyl]-hexahydro-2H-
cyclopentarb~furan-2-one (4):
A solution of anhydrous THF (1.4 L), LiCl (11.7 g,
0.28 mol) and the phosphonate 2 (67.0 g, 0.28 mol) was
cooled to 0C and triethylamine (39.2 mL, 0.28 mol) was
added dropwise. A solution of the aldehyde 3 (68.5 g,
0.25 mol) in dry CH2Cl2 (320 mL) was added dropwise to
the cold suspension and the resulting mixture was
stirred at 0C for 3 h. The reaction mixture was then
poured into S00 mL of 2 M HCl, and layers were
separated. The aqueous layer was extracted with S00 mL
of CH2Cl2. The combined organic layers were washed with
100 mL each of water and brine followed by drying over
MgSO4. Filtration and solvent removal gave a yellow
2138181
solid which was recrystallized from EtOAc to afford
85.8 g (89%) of 4 as a white solid: mp 151-153C; IH NMR
(CDCl3) ô 8.01 (d, J = 2.0 Hz, 2H), 7.65 - 7.40 (m,
3H), 6.70 (dd, J = 12, 6 Hz, lH), 6.35 (d, J = 12 Hz,
lH), 5.32 (m, lH), 5.15 (m, lH), 2.93 (m, 3H), 2.72 -
2.25 (m, 4H), 1.85 - 1.56 (m, 6H), 1.40 - 1.15 (m, 5H).
C: (3aR, 4R, 5R, 6aS)-5-(Benzoyloxy)-4-~(E)-(3S)-3-
cyclohexyl-3-hydroxy-1-propenyl~-hexahydro-2H-
cyclopenta r b~furan-2-one (5):
A solution of CeCl3-7H2O (19.5 g, 52.3 mmol) and
enone 4 (20.0 g, 52.3 mmol) in 150 mL of CH30H and 70
mL of CH2Cl2 was prepared. NaBH4 (1.92 g, 52.3 mmol)
was added in small portions over a period of 5 min.
The resulting mixture was stirred at ambient
temperature for 45 min and then was poured into a
separatory funnel containing 100 mL each of 25% (v/v)
aqueous acetic acid and CH2Cl2. The layers were
separated and the aqueous layer was extracted with 3 X
50 mL of CH2Cl2. The combined organic layers were
washed with sat. NaHCO3 (50 mL), and brine (50 mL), and
then dried (MgSO4). Upon solvent removal, 23.7 g of a
colorless oil containing nearly equal amounts of the
two diastereomeric allyl alcohols was obtained. The
diastereomers were separated by HPLC (40%
EtOAc/hexane), affording 5 (9.34 g (46%), the less
polar component) as a white solid. IH NMR (CDCl3) ~
8.01 (d, J = 8 Hz, 2H), 7.62 - 7.28 (m, 3H), 5.61 (m,
2H), 5.25 (m, lH), 5.08 (m, lH), 3.85 (m, lH), 2.95 -
2.45 (m, 5H), 2.30 (m, 2H), 1.95 - 1.55 (m, 6H), 1.50 -
0.80 (m, 5H).
D: (3aR, 4R 5R 6aS)-4-[(3R)-3-Cyclohexyl-3-
hydroxYpropyl~-hexahydro-5-hydroxy-2H-
cyclopenta[blfuran-2-one (7):
A solution of the allyl alcohol 5 (10.0 g, 26.0
35 mmol) in warm methanol (100 mL) was cooled to ambient
2138181
temperature. Anhydrous K2CO3 (3.6 g, 26.0 mmol) was
added and the resulting mixture was stirred at ambient
temperature for 3 h. The reaction mixture was
concentrated and the residue was partitioned between
100 mL each of EtOAc and 1 M HCl. The layers were
separated and the aqueous phase was extracted with 3 X
50 mL of EtOAc. The combined organic layers were
washed with 50 mL of water, 2 X 50 mL of sat. NaHCO3,
50 mL of brine, and dried over MgSO4. Filtration and
evaporation gave the diol 6 (9.8 g, 92% yield, ~
=0.26, 100% EtOAc), which was used in the subsequent
reaction without further purification.
The crude diol 6 (9.8 g, 26 mmol) was dissolved in
50 mL of EtOAc and a catalytic amount (0.1 g) of 5%
Pd/C was added. This mixture was hydrogenated at 30-40
psi in a Parr hydrogenation apparatus for 3 h and then
filtered through a short pad of Celite. The filtrate
was concentrated and the crude yellow oil was purified
by passage through a short column of silica (~ =0.26,
EtOAc) to afford 7 (5.06 g, 70% yield from 5) as a
colorless, viscous oil which solidified upon standing.
H NMR (CDCl3) ~ 4.95 (m, lH), 4.05 (m, lH), 3.35 (m,
lH), 2.80 (m, lH), 2.58 (m, 2H), 2.30 (m, lH), 2.00 (m,
14H).
E: (3aR, 4R, 5R, 6aS)-4-~(3R)-3-Cyclohexyl- 3-
(tetrahYdropyran-2-yloxy)propyl~-hexahydro-5-
(tetrahydropyran-2-yloxy)-2H-cyclopentarb1furan-2-one
(8):
A solution of the diol 7 (6.0 g, 21.2 mmol) and
dihydropyran (7.80 mL, 84.8 mmol) in CH2Cl2 (100 mL) was
cooled to 0C. A catalytic amount of p-TsOH (0.05 g,
0.26 mmol) was added and the mixture was stirred for 30
min at 0C. The reaction was then quenched by adding
sat. aqueous NaHCO3 (10 mL). The layers were separated
and the aqueous phase was extracted with 2 X 25 mL of
CH2Cl2. The combined organic layers were dried over
21~181
anhydrous K2CO3, filtered and concentrated to afford a
colorless oil which was purified by passage through a
short column of silica (Rf =0.46, 1:1 EtOAc/hexanes).
The bis-THP ether 8 (8.59 g, 89~ yield) was isolated as
a colorless oil which solidified upon standing. IH NMR
(CDCl3) ~ (characteristic peaks only) 5.00 (m, lH),
4.75 - 4.45 (m, 2H), 3.85 (m, 2H), 3.60 - 3.30 (m, 4H).
F: (9S, llR, 15R)-11,15-Bis-(tetrahydropyran-2-yloxy)-
15-cyclohexyl-2,3,4,5,6,16,17,18,19,20-decanor-9-
(triethylsilyloxy)prostanol Triethylsilyl Ether (10):
A suspension of lithium aluminum hydride (1.43 g,38.0 mmol) in 50 mL of anhydrous THF was cooled to 0C
and a solution of the lactone 8 (8.59 g, 19.0 mmol) in
THF (100 mL) was added dropwise. The resulting mixture
was stirred at 0C for 3 h, after which 1.5 mL of H2O,
1.5 mL of 15~ NaOH and 4.5 mL of H2O were sequentially
added. After warming to ambient temperature, 100 mL of
EtOAc was added and the solids were filtered off. The
filter cake was washed thoroughly with 3 X 50 mL of
EtOAc and the filtrates were dried by passage through a
short pad of anhydrous MgSO4. Evaporation afforded 9
(9.02 g) as a colorless oil which was used in the
subsequent step without further purification (Rf =0.31,
80:20 EtOAc/hexanes).
A mixture of the crude diol 9 (9.02 g, 19.0 mmol),
triethylsilyl chloride (9.65 mL, 57.0 mmol),
dimethylaminopyridine (0.41 g, 3.42 mmol),
triethylamine (16.0 mL, 114 mmol) and anhydrous N,N-
dimethylformamide (50 mL) was stirred at ambient
temperature for 14 h under N2. The reaction mixture
was then diluted with 250 mL of CH2Cl2 and the solution
was washed with 3 X 50 mL of H2O. The combined water
washes were extracted with 2 X 50 mL of CH2Cl2. The
organic layers were combined, dried (MgSO4), filtered
and concentrated to afford a yellow oil which was
chromatographed on silica (Rf =0.4, 1:9 EtOAc/hexanes).
2138181
Pure 10 (11.23 g, 86% yield from 8) was obtained as a
slightly yellow oil. lH NMR (CDCl3) ~ (characteristic
peaks only) 4.62 (m, 2H), 4.15 - 3.25 (broad m, 7H),
2.30 - 1.15 (broad m, 18H), 0.95 (broad t, 18H), 0.65
(broad q, 12H).
G: (9S, llR, 15R)-11,15-Bis-(tetrahYdropyran-2-yloxy)-
15-cyclohexyl-2,3,4,5,6,16 17,18,19 20-decanor-9-
(triethylsilyloxy)prostanal (11):
A solution of oxalyl chloride (0.51 mL, 0.57 mmol)
in anhydrous CH2Cl2 (15 mL) was cooled to -78C under N2.
A solution of anhydrous DMSO (0.81 mL, 11.4 mmol) in
CH2Cl2 (2.0 mL) was then added dropwise. After 2 min, a
solution of 10 (2.6 g, 3.8 mmol) in 8 mL of dry CH2Cl2
was introduced dropwise via syringe over a period of 2
min. The resulting mixture was stirred at -78C for 2
h at which time triethylamine (2.7 mL, 19.0 mmol) was
added. The reaction was stirred for 15 min and then
allowed to warm to ambient temperature. The mixture
was partitioned between 100 mL of EtOAc and 10 mL of
H2O and the organic layer was washed with an additional
10 mL of H2O, 10 mL of brine and dried (MgSO4). Solvent
removal gave a yellow oil which was subjected to
chromatography on silica gel (Rf 0.2, 10%
EtOAc/hexanes) to afford 11 (1.4 g, 65% yield) and some
starting material (0.83 g). IH NMR (CDCl3) ~ 9.80
(broad s, lH), 4.62 (m, 2H), 4.20 (m, lHj, 3.85 - 3.60
(m, 3H), 3.40 (m, 3H), 2.80 (m, lH), 2.45 - 2.05 (m,
4H), 1.95 - 1.10 (broad m, 27H), 0.95 (broad t, 9H),
0.55 (broad q, 6H).
H: (5Z)-(9S, llR, 15R~-11,15-Bis-(tetrahydropyran-2-
yloxy)-15-cyclohexyl-2,3,4,16,17,18,19,20-octanor-9-
(triethylsilyloxy)-5-prostenoic Acid Methyl Ester (12):
A solution of 18-crown-6 (8.50 g, 32.1 mmol) and
bis(2,2,2-trifluoroethyl)
(methoxycarbonylmethyl)phosphonate (3.72 g, 11.7 mmol)
213~181
in 110 mL of THF was cooled to -78C. KHMDS (0.5 M in
toluene, 23.4 mL, 11.7 mmol) was added to the above
mixture and the solution was stirred for 15 min.
Aldehyde 11 (6.11 g, 10.7 mmol) in 5.0 mL of THF was
added dropwise over a period of 15 min. The reaction
was stirred at -78C for 2 h, then warmed up to 0C and
stirred at that temperature for 2 more hours. The
reaction was quenched by adding 50 mL of saturated
aqueous NH4Cl and the mixture was allowed to warm to
room temperature. The layers were separated and the
aqueous layer was extracted with 2 X 50 mL of EtOAc.
The combined organic layers were washed with 2 X 50 mL
of brine and dried (K2CO3). Filtration and solvent
removal gave a crude yellow oil which was purified by
passage through a short plug of silica to afford a
mixture of 12 and its E isomer (9:1 ratio, 6.28 g, 95%
yield). The isomers were separated by chromatography
on silica gel (~ =0.56, and 0.47, for the major and
minor isomers respectively, 40% Et2O/hexane); 4.57 g of
pure 12 and 0.97 g of a 1:1 E/Z mixture were isolated.
IH NMR (CDCl3) ~ 6.35 (m, lH), 5.78 (broad d, J = 12.0
Hz, lH), 4.65 (m, 2H), 4.28 (m, lH), 3.90 (m, 2H), 3.70
(s, 3H), 3.55-3.30 (m, 3H), 2.80 (m, 2H), 2.35-2.05 (m,
lH), 2.00-1.10 (broad m, 30H), 0.95 (broad t, 9H), 0.60
(broad q, 6H).
I: (5Z)-(9S, llR, 15R)-11,15-Bis-(tetrahydropyran-2-
yloxy)-15-cyclohexyl-2,3 4,16,17 18,19 20-octanor-9-
(triethylsilyloxy)-5-prosten-1-ol (13):
A solution of 12 (2.0 g, 3.22 mmol) in 20 mL of
anhydrous THF was cooled to 0C under N2. A solution of
diisobutylaluminum hydride (1.5 M in toluene, 6.5 mL,
9.66 mmol) was added dropwise and the resulting mixture
was stirred at 0C for 2 h. The reaction was then
quenched by careful addition of CH30H (5 mL), allowed
to warm up to ambient temperature, and diluted with 50
mL of THF. The resulting cloudy solution was treated
21381~1
with 50 mL of a saturated aqueous solution of sodium
potassium tartrate and the biphasic mixture was stirred
for 1 h. The layers were then separated and the
aqueous layer was extracted with 2 X 50 mL of THF. The
organic extracts were combined, washed with brine (50
mL), and dried (MgSO4). Filtration and solvent removal
gave a pale yellow oil which was purified by
chromatography on silica gel (~ =0.26, 4:6 Et2O/hexane)
to yield 13 (1.95 g, 95% yield) as a colorless oil.
This compound was used immediately in the subsequent
reaction. IH NMR (CDCl3) ~ 5.65 (m, 2H), 4.65 (m, 2H),
4.30-3.25 (broad m, 5H), 2.40-2.05 (broad m, 4H), 2.00-
1.10 (broad m, 32H), 1.00 (broad t, 9H), 0.60 (broad q,
6H).
J: (5Z)-(9S, llR, 15R)-11,15-Bis-(tetrahydropyran-2-
yloxy)-15-cYclohexyl-9-hYdroxy-3-oxa-16,17 18,19,20-
pentanor-5-prostenoic Acid t-Butyl Ester (15):
A mixture of 13 (1.95 g, 3.28 mmol), t-butyl
bromoacetate (5.11 g, 26.24 mmol), tetrabutylammonium
hydrogen sulfate (0.8 g, 2.35 mmol), toluene (45 mL)
and 25% (w/w) aqueous NaOH (30 mL) was stirred
vigorously at room temperature for 18 h. The layers
were separated and the aqueous layer was extracted with
2 X 25 mL of EtOAc. The combined organic extracts were
washed with brine (15 mL), dried (MgSO4), and
concentrated. The crude product was purified by
chromatography on silica gel (~F0.56, 20%
EtOAc/hexane) to yield 2.19 g of 14 (contaminated with
some t-butyl bromoacetate) and 0.48 g of the starting
allyl alcohol 13. The allyl ether 14 thus obtained was
used in the desilylation reaction without further
purification.
The silyl ether 14 (0.5 g) obtained above was
dissolved in 3.0 mL of DMSO and to it was added 2.2 mL
of tetrabutylammonium fluoride (1.0 M in THF, 2.2
mmol). The mixture was stirred at ambient temperature
213~
for 30 min and then partitioned between 50 mL EtOAc and
10 mL of brine. The aqueous layer was extracted with 2
X 10 mL of EtOAc and the combined organic extracts were
dried over MgSO4. Evaporation and chromatography on
silica gel (Rf =0.44, 50% EtOAc/hexane) afforded 0.28 g
of 15 as a colorless oil. IH NMR (CDCl3) ~ 5.65 (m,
2H), 4.62 (m, 2H), 4.16 (m,. lH), 4.10-3.75 (m, 3H),
3.95 (s, 2H), 3.45 (m, 2H), 2.50-0.90 (broad m, 35H),
1.46 (s, 9H); High Resolution CI MS m/z (CI) calcd.
for C34H59O8 (MH+) 595.4209, found 595.4208.
K: (5Z)-(9R, llR 15R)-9-Chloro-15-cyclohexyl-11,15-
dihydroxy-3-oxa-16 17 18,19,20-pentanor-5-prostenoic
Acid t-Butyl Ester (V):
The hydroxyester 15 (0.28 g, 0.47 mmol) was
dissolved in 4.0 mL of a stock solution containing 48.0
mL of CH3CN, 0.79 mL of pyridine, and 0.97 mL of CCl4.
Triphenylphosphine (0.18 g, 0.70 mmol) was added and
the resulting mixture was stirred at ambient
temperature for 17 h. The reaction mixture was treated
with 10 mL of a 1:1 solution of Et2O/hexanes and the
precipitate formed was filtered off. The filtrate was
concentrated and purified by chromatography (silica
gel, Rf =0.47, 40:60 Et2O/hexanes) to yield pure 16 (90
mg, 34%) as a colorless oil.
A solution of 16 (80 mg, 0.13 mmol) in 7.0 mL of
65% (v/v) aqueous acetic acid was heated to 65-70~C for
45 min. The reaction mixture was cooled to room
temperature and concentrated. The resulting residue
was redissolved in anhydrous EtOH and the solvent was
again evaporated. The residue thus obtained was
purified by chromatography on silica gel (~ =0.4,
60:40 EtOAc/hexanes) to yield 60 mg (100%) of V as a
colorless, viscous oil. lH NMR (CDCl3) ~ 5.69 (m, 2H),
4.32 - 3.85 (m, 5H), 3.38 (m, lH), 2.50 - 1.95 (m, 5H),
1.95 - 0.80 (broad m, 29H) 1.43 (s, 9H); l3C NMR (CDCl3)
~ 169.9, 131.7, 126.8, 82.0, 75.6, 75.1, 67.9, 66.6,
2138181
54.2, 51.0, 44.3, 43.7, 31.4, 30.3, 30.1, 29.3, 28.1,
28.0, 26.5, 26.3, 26.1; High Resolution Cl MS m/z
calcd. for C24H42OsCl (MH+) 445.2720, found 445.2716.
18
2138181
E~CAMPLE 2: 8Y~n~:sI8 OF CONPOIJND VI
~O~CO~u~ S,~ o~COz~ O~CO~
~0 ~0 ~0 "
THP~ Tl IPO T~P~ THI~ Pd
t7
a .~=~OAco~ cl ~=~o~
T~ ~0
A: (5Z)-(9S, llR 15~)-11,15-Bis-(tetrahydropyran-2-
yloxy)-15-cyclohexyl-9-hydroxY-3-oxa-16 17 18,19.20-
pentanor-5-prostenoic Acid (17):
Hydroxyester 15 (0.454 g; 0.76 mmol; see Example
1) was dissolved in 10 mL of methanol and 2 mL of
water. Lithium hydroxide monohydrate (0.16 g; 500
mol%) was added and the mixture was stirred at room
temperature. After 18 h, 20 mL of saturated, aqueous
KH2PO4 and 20 mL CH2Cl2 were added, the layers were
separated, and the aqueous phase was washed with
additional CH2Cl2 (3 X 20 mL). The combined organic
layers were dried over Na2SO4, filtered, and
concentrated, affording 0.47 g of a colorless oil which
was used directly in the next reaction.
B: (5Z)-(9S l}R 15R)-11 15-Bis-ttetrahydropYran-2-
yloxy3-15-cyclohexyl-9-hydroxy-3-oxa-16.17 18.19.20-
pentanor-5-prostenoic Acid Isopropyl Ester (18):
Crude acid 17 from above (0.23 g; 0.43 mmol) was
dissolved in 10 mL of acetone. DBU (0.25 mL; 400 mol%)
and isopropyl iodide (0.21 g; 300 mol%) were added and
the mixture was stirred for 12 h at room temperature.
After evaporation, the residue was applied to a silica
gel column and eluted with hexane/EtOAc, 1/1, to afford
0.157 g (63%) of isopropyl ester 18 as a colorless oil
19
2I3Sl~l
(Rf -0.49). lH NMR (CDCl3) ~ (characteristic peaks
only) 5.80-5.52 (m, 2H), 5.15 (sep, lH, J=6.2 Hz), 4.03
(broad s, 2H), 1.27 (d, 6H, J=6.2 Hz).
C: (5Z)-(9R llR 15R)-9-Chloro-15-cyclohexY~ l5
dihYdroxY-3-oxa-16.17 18 19 20-pentanor-5-~rostenoic
Acid IsopropYl Ester (VI):
The hydroxyester 18 (0.146 g; 0.25 mmol) was
dissolved in 3.0 mL of a stock solution containing 48
mL of CH3CN, 0.79 mL of pyridine, and 0.97 mL of CC14.
Triphenylphosphine (0.10 g; 150 mol%) was added and the
resulting mixture was stirred at room temperature for
17 h. The reaction mixture was treated with 10 mL of a
1:1 solution of Et2O/hexanes and the precipitate was
filtered off. The filtrate was concentrated and
chromatographed on silica gel (hexane/EtOAc, 4/1),
affording 0.108 g of a colorless oil which consisted of
a nearly equimolar mixture of desired chlorinated
material 19 with its undesired 5,8-diene elimination
product.
A solution of crude l9 from above in 10 mL of 65%
(v/v) aqueous acetic acid was warmed to 65C for 45
min. The mixture was then cooled to room temperature
and concentrated. The resulting residue was then
purified by silica gel chromatography (hexane/EtOAc,
2/3), affording 27 mg (25% based on 18) of pure VI (~
=0.56) as a colorless oil, and 69 mg of a mixture of VI
and its 5,8-diene elimination product (~ =0.45). IH
NMR (CDCl3) ~ 5.67 (m, 2H), 5.08 (sep, lH, J=6.1),
4.30-3.95 (m, 6H), 3.40 (m, lH), 2.35 (m, 2H), 2.30-
2.00 (m, 3H), 1.93-1.35 (m, 12H), 1.25 (d, 6H, J=6.2
Hz), 1.22-0.90 (m, 6H); l3C NMR (CDCl3) ~ 170.2, 131.8,
126.7, 75.7, 75.2, 68.8, 67.6, 66.7, 61.2, 54.2, 51.1,
44.4, 43.6, 31.4, 30.2, 30.1, 29.3, 28.0, 26.5, 26.3,
26.1, 21.8; High Resolution CI MS m/z calcd. for C23
H40OsCl (MH+) 431.2564, found 431.2569.
21381~1
E~P~B 3: 8YNTHB~I8 OF COMPOIJND VII
o a~
~ ~ V~
A: (5Z)-(9S llR, lSR)-11 15-Bis-(tetrahYdropYran-2-
ylox~)-15-cyclohexyl-9-hydroxy-16 17,18,19 20-pentanor-
5-prostenoic Acid Isopropyl Ester (22):
To a solution of 5.0 g (11 mmol) of lactone 8 (see
Example 1) in 40 mL of THF at -78C was added dropwise
9.6 mL (14.4 mmol) of a 1.5 M solution of
diisobutylaluminum hydride in toluene. After 1.5 h, 5
mL of MeOH was added, the mixture was stirred for 10
min at -78C and then warmed to room temperature. This
solution was then added to a mixture of 20 mL of
saturated, aqueous NH4Cl, 35 mL of EtOAc, and 35 mL of
saturated, aqueous sodium potassium tartrate. The
mixture was stirred for 20 min, layers were separated,
and the agueous layer was washed with EtOAc (3 X 40
mL). The combined organic layers were dried over
MgSO4, filtered, and evaporated. The resulting residue
was purified by silica gel chromatography
(EtOAc/hexane, 1/1), affording 4.5 g (90%) of lactol 20
which was used directly in the next reaction.
To a solution of 14.1 g (31.8 mmol) of (1-
carboxypent-5-yl)triphenyl-phosphonium bromide in 100
mL of THF at 0C was added dropwise 59 mL (59 mmol) of
a 1 M solution of potassium t-butoxide in THF. After
20 min, 4.5 g (9.9 mmol) of lactol 20 in 20 mL of THF
was added dropwise. The reaction was quenched after 2
h by pouring into 150 mL of a 1/1 (v/v) mixture of
EtOAc/saturated, aqueous NH4Cl . The layers were
21
213~181
separated and the aqueous layer was extracted with
EtOAc (3 X 70 mL). The combined organic layers were
dried over MgSO4, filtered and evaporated, leaving 7.6
g of crude acid 21 as an oil.
Crude acid 21 (7.6 g) was dissolved in 55 mL of
acetone, cooled to 0C, and 8.6 g (56 mmol) of DBU was
added dropwise. The reaction was warmed to room
temperature and 8.5 g (50 mmol) of isopropyl iodide was
added dropwise. After stirring for 14 h, the mixture
was poured into 100 mL of a 1/1 (v/v) mixture of
EtOAc/saturated, aqueous NH4Cl. The layers were
separated and the aqueous phase was extracted with
additional EtOAc (2 X 100 mL). The combined organic
layers were dried over MgSO4, filtered, evaporated, and
chromatographed on silica gel (EtOAc/hexane, 2/3)
affording 1.98 g (35% from lactol 20) of 22. IH NMR
(CDCl3) ~ (characteristic peaks only) 5.58-5.28 (m,
2H), 4.97 (sep, J=6.2 Hz, lH), 1.1 (d, J=6.2 Hz, 6
H).
B: (5Z)-(9R llR 15R)-9-Chloro-15-cyclohexyl-11,15-
dihydroxy-16,17 18,19,20-pentanor-5-prostenoic Acid
Isopropyl Ester (VII):
Hydroxyester 22 (513.1 mg; 0.8864 mmol) was
dissolved in 6.6 mL of a stock solution containing 48.0
mL of CH3CN, 0.79 mL of pyridine, and 0.97 mL of CCl4.
Triphenylphosphine (348.8 mg; 150 mol%) was added and
the mixture was stirred for 45 h at room temperature.
The reaction was then diluted with 7 mL of Et2O and 14
mL of hexane. After stirring for 10 min, solids were
filtered off and the filtrate was evaporated. The
resulting solids were triturated three times with 15 mL
of hexane/Et2O (1/2). Combined hexane/Et2O washes were
concentrated down to 0.75 g of a white solid which was
then redissolved in hexane/Et2O and chromatographed on
silica gel. Elution with hexane/Et2O, 5/1, afforded
21381~1
372.8 mg of semipure 23 which was used directly in the
next reaction.
Crude 23 from above was dissolved in 20 mL of 65
(v/v) aqueous HOAc and warmed to room temperature.
After 1.5 h, the reaction was concentrated, 15 mL of
H2O was added, and the solution was reconcentrated.
Absolute EtOH (15 mL) was added followed, again, by
reconcentration. The resulting oil was purified by
silica gel chromatography (hexane/EtOAc, 2/1),
affording 244.1 mg of a mixture of VII contaminated
with an approximately equal quantity of the 5,8-diene
side product. An 8.2 mg sample was then further
purified by reverse phase HPLC, giving 4.4 mg of pure
VII as a clear, viscous oil. IH NMR (CDCl3) ~ 5.47 (m,
J=8.5 Hz, 2H),-5.01 (sep, J=6.3Hz, lH), 4.10 (dt,
J=4.0, 6.2 Hz, lH), 4.04 (q, J=7.6 Hz, lH), 3.37 (m,
lH), 2.35-2.24 (m, 4H), 2.20-2.07 (m, 4 H), 1.82 (br s,
2H), 1.80-1.50 (m, 13H), 1.36-0.96 (m, 12H). ~3C NNR
(CDCl3) ~ 173.2, 131.0, 126.8, 76.2, 76.0, 67.5, 60.8,
54.3, 51.7, 44.5, 43.5, 34.0, 31.7, 30.0, 29.2 (two
overlapping resonances), 27.9, 26.6, 26.4, 26.2, 26.1,
24.9, 21.8. High Resolution CI MS m/z calcd. for
C24H42O4Cl (MH+) 429.2772, found 429.2763.
EXANPLE 4: 8YNTHE8I8 OF CONPOUND VIII
v~
213S181
A: (5Z)-(9R, llR, 15R)-11-(t-ButyldimethYlsiloxy)-9-
chloro-15-cyclohexyl-15-hydroxy-3-oxa-16,17,18,19,20-
pentanor-5-prostenoic acid t-butyl ester (24):
To a mixture of 127 mg (0.285 mmol) of Compound V
(Example 1), 49 mg (0.72 mmol) of imidazole, 10 mg
(0.082 mmol) of 4-(dimethylamino)pyridine (DMAP), and 5
mL of CH2Cl2 was added 90 mg (0.59 mmol) of tert-
butyldimethylsilyl chloride. After stirring overnight,
10 mL of saturated NH4Cl was added, the mixture was
extracted with CH2Cl2 (3 X 10 mL), the combined organic
layers were dried over MgSO4, filtered and
concentrated, and the residue was chromatographed on
silica gel (20% ethyl acetate in hexane) to afford 87
mg (55%) of 24.
B: (5Z)-(9R, llR, 15R)-11-(t-Butyldimethylsiloxy)-9-
chloro-15-cyclohexyl-15-methoxy-3-oxa-16,17,18,19,20-
pentanor-5-prostenoic acid t-butyl ester (25):
A mixture of 80 mg (0.14 mmol) of 24, 100 mg (0.52
mmol) of 2,6-di-t-butylpyridine, 80 mg (0.51 mmol) of
methyl trifluoromethanesulfonate, and 2 mL of CH2Cl2 was
refluxed overnight. The reaction was cooled to room
temperature, poured into 10 mL of saturated NaHC03,
extracted with CH2Cl2 (3 X 10 mL), the combined organic
layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel (10% ethyl acetate in hexane) to afford 35
mg (44%) of 25.
C: (5Z)-(9R, llR, 15R)-9-Chloro-15-cyclohexYl-ll-
hydroxy-15-methoxy-3-oxa-16,17,18,19,20-pentanor-5-
prostenoic acid t-butyl ester (VIII):
To a mixture of 32 mg (0.056 mmol) of 25 and 1.5
mL of THF was added 0.12 mL (0.12 mmol) of 1 M
tetrabutylammonium fluoride (TBAF) in THF. After 30
min, 4 mL of saturated NH4Cl was added, the mixture was
extracted with ethyl acetate (3 X 5 mL), the combined
24
21~81~1
organic layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel (40% ethyl acetate in hexane) to afford 24
mg (93%) of VIII. 13C NMR (CDCl3) ~ 169.77 (C), 130.90
(CH), 127.43 (CH), 85.89 (CH), 81.69 (C), 76.05 (CH),
67.83 (CH2), 66.56 (CH2), 61.00 (CH), 57.83 (CH3), 54.06
(CH), 51.92 (CH), 44.45 (CH2), 40.65 (CH), 29.92 (CH2),
29.83 (CH2), 29.02 (CH2), 28.42 (CH3), 28.10 (CH2), 26.64
(CH2), 26.37 (CH2). CI MS, m/z calcd. for C25H~O5Cl
(MH+) 459.2877, found 459.2877.
EXAMPLB 5: 8YNTHE8I8 OF CONPOUND IX
~ ~
A: (5E)-(llR 15R~-15-CyclohexYl-11.15-dihYdroxY-3-
oxa-16 17 18 19 20-pentanor-5-prostenoic acid isopropyl
ester (IX):
To a solution of the ester VI (390 mg, 0.90 mmol)
(see example 2) and AIBN (10 mg) in anhydrous toluene
(9.0 mL) was added n-Bu3SnH (0.47 mL, 1.80 mmol). The
resulting mixture was heated at reflux for 4 h. The
solvent was evaporated and the residue was applied to a
silica gel column for purification. The product IX
( ~ 0.4, 60% EtOAc/hexane) was isolated as a colorless
oil (340 mg isolated, 95% yield). IH NMR (CDCl3) ~
5.60 (m, 2H), 5.05 (septet, J = 6.5 Hz, lH), 3.98 (m,
4H), 3.85 (m, lH), 3.30 (m, lH), 2.22 (m, lH), 2.00 (m,
lH), 1.80 - 0.85 (broad m, 23H), 1.21 (d, J = 6.4 Hz,
6H); C NMR (CDCl3) ~ 170.11, 134.59, 126.54, 79.15,
76.35, 72.07, 68.50, 67.18, 53.57, 44.21, 43.62, 38.21,
34.19, 32.05, 29.47, 29.32, 28.94, 27.98, 26.56, 26.36,
26.21, 21.84.
213~181
EXAMPLE 6: SY~.-n~;8IS OF COMPOIJND X
HO dH l~d ~ ~ ~T~
~
26
2138181
A: (3aR, 4R 5R, 6aS)-4-[(E)-(3S)-3-Cyclohexyl-3-
(tetrahydropyran-2-yloxy)Propenyll-hexahydro-5-
(tetrahydropyran-2-yloxy)-2H-cyclopenta[b~furan-2-one
(26):
To 15.7 g (55.9 mmol) of diol 6 (Example 1) in 120
mL of CH2Cl2 at 0C was added 12.0 g (142 mmol) of 3,4-
dihydro-2H-pyran (DHP) and 520 mg (2.7 mmol) of
p-toluenesulfonic acid monohydrate (pTSA). After 1 h
at 0C, 100 mL of saturated NaHCO3 was added, the
mixture was extracted with CH2C12 (3 X 100 mL), the
combined organic layers were dried over MgSO4, filtered
and concentrated and the residue was chromatographed on
silica gel (40% ethyl acetate in hexane) to afford
13.3 g (53%) of bis-THP ether 26.
B: (13E)-(9S~llR~155)-11~15-Bis(tetrahydropYran-2
yloxy)-15-cYclohexyl-2~3~4~5~6~l6~l7~l8~l9~2o-decanor
9-hydroxy-13-prostenol (27):
To a solution of 15.0 g (33.4 mmol) of 26 in 150
mL of THF at 0C was added 53 mL (80 mmol) of 1.5 N
solution of diisobutylaluminum hydride (DIBAL-H) in
toluene. After 3 h the reaction was poured into 300 mL
of 1:1 ethyl acetate:saturated sodium potassium
tartrate and stirred for 1 h. The layers were
separated, the aqueous layer was extracted with ethyl
acetate (3 X 100 mL), the combined organic layers were
dried over MgSO4, filtered and concentrated to afford
14.89 g (98%) of crude diol 27.
C: (13E)-(9S llR 15S)-11 15-Bis(tetrahydropyran-2-
yloxy)-15-cYclohexyl-2~3~4~5~6~l6~l7~l8~l9~2o-decanor
9-(triethylsiloxy)-13-prostenol triethylsilyl ether
(28):
To a mixture of 14.8 g (32.7 mmol) of 27, 5.94 g
(87.4 mmol) of imidazole, 0.44 g (3.6 mmol) of DMAP,
and 150 mL of CH2Cl2 was added 11.5 g (76.3 mmol) of
triethylsilyl chloride. After 5 h, 150 mL of saturated
2138181
NH4Cl was added, the layers were separated, and the
aqueous layer was extracted with CH2Cl2 (2 X 100 mL).
The combined organic layers were dried over MgSO4,
filtered, and concentrated, and the residue was
chromatographed on silica gel (10% ethyl acetate in
hexane) to afford 21.9 g (100%) of bis-silyl ether 28.
D: (13E)-(9S,llR,155)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-2,3,4,5,6,16,17,18,19,20-decanor-
9-(triethylsiloxy)-13-prostenal (29):
To 12.5 g (98.6 mmol) of oxalyl chloride in 150 mL
of CH2Cl2 at -78C was added dropwise a solution of DMSO
(13.0 g, 166 mmol) in 15 mL of CH2Cl2. After 30 min, a
solution of 28 (22.4 g, 32.9 mmol) in 60 mL of CH2Cl2.
After 5 h, 36 g (360 mmol) of NEt3 was added, and the
reaction was stirred for 30 min at -78C and then at
room temperature for 30 min. The mixture was poured
into 200 mL of saturated NH4Cl and extracted with CH2Cl2
(3 X 150 mL), the combined organic layers were dried
over MgSO4, filtered, and concentrated, and the residue
was chromatographed on silica gel (20% ethyl acetate in
hexane) to afford 18.3 g (99%) of aldehyde 29.
E: (5Z, 13E)-(9S,llR, 155)-11,15-Bisftetrahydropyran-
2-yloxy)-15-cyclohexyl-2,3,4,16,17,18,19,20-octanor-9-
(triethylsiloxy)-5,13-prostadienoic acid methyl ester
(30):
To a mixture of 16.5 g (51.9 mmol) of bis(2,2,2-
trifluoroethyl) (methoxycarbonylmethyl)phosphonate,
28.2 g (107 mmol) of 18-crown-6, and 200 mL of THF at
-78C was added dropwise 85 mL (42.5 mmol) of 0.5 N
KHMDS in toluene. After 30 min, a solution of 18.3 g
(32.4 mmol) of 29 in 50 mL of THF was added dropwise
and the reaction stirred for 2 h. The mixture was
poured into 150 mL of saturated NH4Cl, extracted with
ethyl acetate (3 X 100 mL), the combined organic layers
were dried over MgSO4, filtered and concentrated, and
28
213~
the residue was chromatographed on silica gel (15%
ethyl acetate in hexane) to afford 9.78 g (49%) of
crotonate 30, as well as 3.58 g (18%) of a mixture of
olefin cis: trans isomers.
F: (5Z, 13E)-(9S,llR, 15S)-11,15-Bis(tetrahydropyran-2-
Yloxy)-15-cyclohexyl-2,3,4,16,17,18,19,20-octanor-9-
(triethylsiloxy)-5,13-prostadienol (31):
To a solution of 9.11 g (14.7 mmol) of 30 in 40 mL
of THF at 0C was added dropwise 24 mL (36 mmol) of a
1.5 M solution of DIBAL-H in toluene. After 1 h, the
reaction was added to 100 mL of saturated NH4Cl,
extracted with ethyl acetate (3 X 75 mL), the combined
organic layers were dried over MgSO4, filtered, and
concentrated to afford 8.7 g (100%) of crude allyl
alcohol 31.
G: (5Z, 13E)-(9S,llR, 15S)-11,15-Bis(tetrahYdropyran-2-
yloxY)-l5-cyclohexyl-9-hydroxy-2~3~4~l6~l7~l8~l9~2
octanor-5,13-prostadienol (32):
To a solution of 8.7 g (14.7 mmol) of 31 in 60 mL
of THF at 0C was added 20 mL (20 mmol) of a 1 M
solution of TBAF in THF. After lh, 75 mL of saturated
NH4Cl was added, the mixture was extracted with ethyl
acetate (3 X 75 mL), the combined organic layers were
dried over MgSO4, filtered, and concentrated, and the
residue was chromatographed on silica gel (3/2 ethyl
acetate/hexane) to afford 3.79 g (54%) of diol 32.
H: (5Z, 13E)-(9S,llR, 15S)-11,15-Bis(tetrahydropyran-
2-yloxy)-15-cyclohexyl-9-hYdroxy-3-oxa-16,17,18,19,20-
pentanor-5,13-prostadienoic acid isopropyl ester (33):
A mixture of 32 (3.76 g, 7.7 mmol), NaOH (6.1 g,
150 mmol), water (30 mL), toluene (30 mL),
tetrabutylammonium hydrogen sulfate (1.05 g, 3.1 mmol),
and isopropyl bromoacetate (3.63 g, 20.1 mmol) at room
temperature was vigorously stirred for 30 min. The
29
2~3~181
layers were separated, the aqueous layer was extracted
with ethyl acetate (2 X 30 mL), the combined organic
layers were dried over MgSO4, filtered and
concentrated, and the residue was chromatographed on
silica gel (40% ethyl acetate in hexane) to afford
ester 33 (3.06 g, 69%).
I: (5Z 13E)-(9R llR 15S)-11 15-Bis(tetrahydropyran-2-
yloxY)-9-chloro-15-cyclohexyl-3-oxa-16 17 18 19 20-
pentanor-5 13-prostadienoic acid isopropyl ester (34):
To a mixture of 33 (3.0 g, 5.2 mmol) and 30 mL of
pyridine at 0C was added methanesulfonyl chloride
(1.48 g, 12.9 mmol). The reaction was brought to room
temperature after 15 min, stirred for an additional 1.5
h, and poured into a suspension of Bu4NCl (14.5 g, 52.2
mmol) in 45 mL of toluene. The mixture was stirred at
room temperature overnight, at 55-60C for 2 h, poured
into 100 mL of saturated NH4Cl, extracted with ethyl
acetate (3 X 75 mL), the combined organic layers were
dried over MgSO4, filtered, and concentrated, and the
residue was chromatographed on silica gel (30% ethyl
acetate in hexane) to afford 3.0 g of an oil which
consisted of a mixture of chlorinated compound 34 and
the 8,9-olefin elimination by-product. This mixture
was used in the next step without further purification.
J: (5Z 13E)-(9R llR 15S)-9-Chloro-15-cyclohexyl-
11 15-dihydroxy-3-oxa-16 17 18 19 20-pentanor-5 13-
prostadienoic acid isopropyl ester (35):
The sample from above (3.0 g) was dissolved in 40
mL of isopropyl alcohol and 5 mL of water was added,
followed by 2.3 mL of 12 M HCl. After 3 h, saturated
NaHC03 was added (50 mL), the mixture was extracted
with ethyl acetate (3 X 50 mL), the combined organic
layers were dried over MgS04, filtered, and
concentrated, and the residue was purified by radial
chromatography (20/1 toluene/isopropanol) to afford 324
21381~1
mg of pure chloride 35 (32%), as well as 278 mg (28~,
calculated as the chloride) of a mixture of 35 with the
8,9-olefin. ~3C NMR (CDCl3) ~ 170.04 (C), 134.51 (CH),
132.64 (CH), 130.43 (CH), 127.58 (CH), 77.34 (CH),
75.40 (CH), 68.60 (CH), 67.60 (CH2), 66.64 (CH2), 59.64
(CH), 55.95 (CH), 53.36 (CH), 43.61 (CH), 43.57 (CH2),
28.83 (CH2), 28.70 (CH2), 26.49 (CH2), 26.06 (CH2), 25.99
(CH2), 21.82 (CH3). CI MS, m/z calcd. for C23H38O5Cl
(MH+) 429.2408, found 429.2408.
K: (5E, 13E)-(llR, 15S)-15-CyclohexYl-11 15-dihydroxy-
3-oxa-16 17,18 19 20-pentanor-5,13-prostadienoic acid
isopropYl ester (X):
A mixture of 35 (60 mg, 0.14 mmol), benzene (3
mL), AIBN (10 mg), and Bu3SnH (90 mg, 0.31 mmol) was
deoxygenated by purging with N2 for 15 min, and heated
at reflux for 1 h. The reaction was concentrated and
chromatographed on silica gel (3:2 ethyl
acetate:hexane) to afford 43 mg (78~) of dechlorinated
product X 13C NMR (CDCl3) ~ 169.98 (C), 134.27 (CH),
134.22 (CH), 133.94 (CH), 126.62 (CH), 77.76 (CH),
77.72 (CH), 71.92 (CH2), 68.41 (CH), 67.06 (CH2), 58.01
(CH), 43.33 (CH), 42.18 (CH), 36.99 (CH2), 32.02 (CH2),
28.94 (CH2), 28.72 (CH2), 27.38 (CH2), 26.48 (CH2), 26.03
(CH2), 25.96 (CH2), 21.74 (CH3). CI MS, m/z calcd. for
C23H39O5 (MH+) 395.2798, found 395.2798.
B2CAMPLE: 7: 8YIITHE:8I~ OF COMPOlrND XI
cl~ c~ 20H
11~ ~
2138181
(5Z)-(9R llR. 15R)-9-Chloro-15-cYclohexyl-11.15-
dihydroxy-3-oxa-16,17,18,19,20-pentanor-5-prostenol
(XI):
A solution of the ester 16 (150 mg, 0.24 mmol;
Example 1) in THF (5.0 mL) was cooled to 0C and to it
DIBAL-H (1.5 M in toluene, 0.5 mL, 0.72 mmol) was added
and the resulting mixture was stirred at 0C for 2.5 h.
The reaction was carefully quenched with a saturated
solution of potassium sodium tartrate (10 mL) and the
biphasic mixture was stirred at ambient temperature for
lh. The organic layer was then separated and the
aqueous layer was extracted with EtOAc (5 X 10 mL).
The combined organic layers were dried (Na2SO4),
filtered, and concentrated to afford a pale yellow
liquid which was purified by chromatography on silica
gel. The intermediate alcohol (RfO.15, 30%
EtOAc/hexanes) was isolated (114 mg, 87% yield) as a
colorless oil and was used in the subsequent reaction.
The above alcohol (54 mg, 0.09 mmol) was mixed
with 65% acetic acid/water (15 mL) and then heated at
70C for 1 h. The solvent was evaporated and the crude
was applied to a column of silica gel for purification.
The triol XI (RfO.15 EtOAc) was isolated as a
colorless oil (33 mg, 88% yield). 1H NMR (CDC13) ~ 5.68
(m, 2H), 4.08 (m, 4H), 3.74 (m, 2H), 3.57 (m, 2H), 3.42
(m, lH), 2.37 (m, 2H), 2.35-1.90 (m, 4H), 1.85-0.90
(broad m, 18H); l3C NMR (CDCl3) ~ 131.06, 127.52, 75.80,
75.54, 71.94, 66.38, 61.82, 60.82, 54.05, 50.87, 44.69,
43.60, 31.29, 29.71, 29.46, 29.22, 28.08, 26.48, 26.28,
26.14; CI MS calcd. for C20H36O4Cl (MH+) 375.2302, found
375.2299.
EXAMPLE 8: S~..n~8I8 OF COMPOUND XII
c~ ~o~co~l cl~ O~c~
~0 ~0
3~ XJI
(5Z, 13E) - (9R.llR. 155)-9-Chloro-15-cvclohexlYl-11.15-
dihydroxy-3-oxa-16.17.18.19.20-pentanor-5,13-
prostadienol (XII):
S To a solution of ester 36 (41 mg, 0.093 mmol)
(German patent DE 3724189) in 5 mL of THF at 0C was
added a 1.5 M solution of DIBAL-H (0.7 mL, 1.05 mmol).
After warming to room temperature and stirring for 1.5
h, 15 mL of saturated NH4Cl was added, the mixture was
extracted with ethyl acetate (3 X 15 mL), the combined
organic layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel (ethyl acetate) to afford triol XII (21 mg,
68%). IC NMR ~CDCl3) ~ 134.59 (CH), 133.00 (CH),
129.61 (CH), 128.21 (CH), 77.56 (CH), 75.16 (CH), 71.54
(CH2), 67.93 (CH2), 66.46 (CH2), 61.76 (CH2), 59.49 (CH),
55.85 (CH), 53.23 (CH), 43.43 (CH), 28.81 (CH2), 28.56
(CH2), 26.46 (CH2), 26.02 (CH2), 26.94 (CH2), 25.57
(CH2). CI MS, m/z calcd. for C20H34O4Cl (MH+) 373.2146,
found 373.2101.
2138181
E~ANP~E 9: 8YNTHE8I8 OF CONPO~ND XIII
O OH
2~
~CO-M _ a d~
~ 4.
A: (3aR. 4R, 5R. 6aS)-4-~(E)-(35)-3-CYclohexyl-3-
(tetrahydropyran-2-yloxv)~ropenvl]-hexahydro-5-
(tetrahydropyran-Z-yloxy)-2H-cYclopentarb~furan-2-ol
(37~:
To a solution of lactone 26 (5.7 g, 12.7 mmol;
Example 6) in 40 mL of THF at -78C was added dropwise
a 1.5 M solution of DIBAL-H in toluene (11.5 mL, 17.2
mmol). After 2 h, the reaction was poured into 70 mL
of a saturated solution of sodium potassium tartrate
and was stirred for 30 min. The mixture was extracted
with ethyl acetate (3 X 50 mL), the combined organic
layers were dried over MgSO4, filtered and
concentrated, and the residue was chromatographed on
silica gel (1/1 hexane/ethyl acetate) to afford 4.7 g
(82%) of lactol 37.
B: (5E. 13E)-(9S. llR. 155)-11.15-Bis(tetrahydro~Yran-
2-YloxY)-15-cyclohexyl-9-hydroxy-2,3,4,16,17,18,19,20-
octanor-5.13-prostadienoic acid methyl ester (38):
A mixture of 37 (5.1 g, 11.3 mmol), Ph3PCH=CO2Me
213~181
(6.6 g, 19.7 mmol), CH2Cl2 (50 mL), and acetic acid (8
drops) was stirred overnight at room temperature. The
mixture was concentrated and chromatographed on silica
gel (1/1 hexane/ethyl acetate) to afford 5.7 g (99%) of
trans-crotonate 38.
C: (5E 13E)-(9S llR, 15S)-11 15-Bis(tetrahYdropyran-
2-yloxY)-9-(t-butYldimethylsiloxY)-15-cyclohexyl-
2 3 4 16,17 18 19 20-octanor-5 13-prostadienoic acid
methYl ester (39):
A mixture of 38 (5.7 g, 11.6 mmol), CH2Cl2 (150
mL), imidazole (1.46 g, 21.5 mmol), DMAP (500 mg, 4.1
mmol), and t-butyldimethylsilyl chloride (2.54 g, 16.9
mmol) was stirred for 1 h, 50 mL of saturated NH4Cl was
added, the layers were separated, the aqueous layer was
extracted with CH2Cl2 (2 X 50 mL), the combined organic
layers were dried over MgSO4, filtered and
concentrated, and the residue was chromatographed on
silica gel (20% ethyl acetate in hexane) to afford 39
(6.05 g, 84%).
D: (5E 13E)-(9S, 11~ 155)-11 15-Bis(tetrahydropyran-
2-yloxy)-9-(t-butyldimethylsiloxy)-15-cyclohexyl-
2 3 4 16 17 18 19 20-octanor-5 13-prostadienol (40):
To a solution of 39 (6.0 g, 9.8 mmol) in THF (50
mL) at 0C was added dropwise a 1.5 M solution of
DIBAL-H in toluene (16 mL, 24 mmol). The reaction was
brought to room temperature and was stirred for 2 h, 75
mL of a saturated solution of sodium potassium tartrate
was added, and the mixture was stirred for 35 min. The
layers were separated, the aqueous layer was extracted
with ethyl acetate (2 X 50 mL), the combined organic
layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel (25% ethyl acetate in hexane) to afford 40
(4.28 g, 74%).
213~181
E: (5E. 13E)-(9S, llR. 155)-11.15-Bis(tetrahydropyran-
2-yloxy)-9-(t-butyldimethylsiloxy)-15-cyclohexyl-3-oxa-
16.17.18,19,20-pentanor-5,13-prostadienoic acid t-butyl
ester (41):
A mixture of 40 (2.4 g, 4.1 mmol), water (25 mL),
toluene (30 mL), NaOH (3.8 g, 95 mmol), Bu4NHSO4 (300
mg, 0.88 mmol), and t-butyl bromoacetate (5.0 g, 25.6
mmol) was stirred vigorously overnight. The layers
were separated, the aqueous layer was extracted with
ethyl acetate (2 X 50 mL), the combined organic layers
were dried over MgSO4, filtered and concentrated, and
the residue was chromatographed on silica gel (20%
ethyl acetate in hexane) to afford 41 (1.48 g, 48%).
F: (5E 13E)-(9S llR 15S)-11.15-Bis(tetrahydropyran-
2-Yloxy)-15-cyclohexYl-9-hydroxy-3-oxa-16 17 18 19 20-
pentanor-5,13-prostadienoic acid t-butyl ester (42):
A mixture of 41 (1.4 g, 2.0 mmol), THF (20 mL), and a 1
M solution of TBAF in THF (6 mL, 6 mmol) was stirred
for 2 h, saturated NH4Cl was added (30 mL), the layers
were separated, the aqueous layer was extracted with
ethyl acetate (2 X 40 mL), the combined organic layers
were dried over MgSO4, filtered, and concentrated, and
the residue was chromatographed on silica gel (1/1
hexane/ethyl acetate) to afford 42 (0.45 g, 38%).
G: (5E 13E)-(9R llR. 15S)-11 15-Bis(tetrahydropyran-
2-yloxy)-9-chloro-15-cyclohexyl-3-oxa-16 17 18 19 20-
pentanor-5 13-prostadienoic acid t-butyl ester (43):
A mixture of 42 (430 mg, 0.72 mmol), PPh3 (350 mg,
1.34 mmol), CH3CN (6 mL), pyridine (112 mg, 1.42 mmol),
and CCl4 (240 mg, 1.55 mmol) was stirred overnight.
The reaction was concentrated and the residue was
chromatographed on silica gel (20% ethyl acetate in
hexane) to afford 43 as a mixture with the
corresponding 8,9-olefin (362 mg, 82% calculated as the
36
21381~1
chloride). This mixture was separated in the next
step.
H: (5E, 13E)-(9R, llR, 15S)-9-Chloro-15-cyclohexyl-
11,15-dihydroxy-3-oxa-16,17,18,19,20-pentanor-5,13-
prostadienoic acid t-butyl ester (XIII):
A mixture of 43 (310 mg, 0.51 mmol calculated as
the chloride), THF (5 mL), water (1 mL), and acetic
acid (9 mL) was heated at 65C for 1 h. The reaction
was concentrated and the residue was chromatographed on
silica gel (4/1 ethyl acetate/hexane) to afford XIII as
a mixture with the corresponding 8,9-olefin (188 mg,
83% calculated as the chloride). The mixture was
separated by reverse-phase HPLC to afford pure XIII (58
mg, 26% from alcohol 46). 13C NMR (CDCl3) ~ 169.67 (C),
134.67 (CH), 133.09 (CH), 131.18 (CH), 128.56 (CH),
81.62 (C), 77.31 (CH), 75.04 (CH), 71.63 (CH2), 67.59
(CH2), 59.38 (CH), 56.34 (CH), 53.08 (CH), 43.32 (CH),
43.38 (CH2), 33.88 (CH2), 28.87 (CH2), 28.77 (CH2), 28.10
(CH3), 26.48 (CH2), 26.05 (CH2), 25.97 (CH2). CI MS m/z
calcd. for C24H40O5Cl 445.2535, found 445.2574.
~13~
EXAMPLE 10: 8YN~HE8I8 OF COMPOIJND VIV
'~ ~ ~
I
s~ sl s~
ndC a~rH ' ~;=~
S~ s~ s~ ~3
n DV
38
21381~1
.
A: 3aR 4R. 5R 6aS)-5-(Benzoyloxy)-4-r(E)-(3R)-3-
cyclohexyl-3-hydroxy-1-propenyll-hexahydro-2H-
cyclopentarb]furan-2-one (44):
To a solution of enone 4 (150 g, 392 mmol; Example
1), cerium trichloride heptahydrate (152 g, 408 mmol),
methanol (500 mL), and CH2Cl2 (1.5 L) at 0C was added
NaBH4 (14.4 g) in 0.2 g portions over 1 h. After
stirring for 2 h, the reaction was poured into 1 M HCl
(500 mL), the layers were separated, the aqueous layer
was extracted with CH2Cl2 (2 X 300 mL), and the combined
organic layers were dried over MgS04, filtered, and
concentrated to provide 150.4 g (100%) of a 1:1 mixture
of the two diastereomeric allylic alcohols. Separation
of 200 g of the mixture (from several combined
reactions) by HPLC (40% ethyl acetate in hexane)
afforded 23.7 g (12%) of alcohol 44.
B: (3aR 4R 5R 6aS)-4-~(E)-(3R)-3-Cyclohexyl-3-
hydroxy-1-propenyl]-hexahydro-5-hydroxy-2N-
cyclopentarb]furan-2-one (45):
To a solution of 44 (7.35 g, 19.1 mmol) in
methanol (100 mL) was added K2CO3 (2.64 g, 19.1 mmol).
After 2 h, 200 mL of 2 M HCl was added, the mixture was
extracted with ethyl acetate (3 X 100 mL), and the
combined organic layers were dried over MgSO4,
filtered, and concentrated to afford crude diol 45
(5.53 g), which was used in the next step without
purification.
C: (3aR, 4R, 5R 6aS)-4-r(3S)-3-Cyclohexyl-3-
hydroxypropyl~-hexahydro-5-hydroxy-2H-
cyclopenta~b]furan-2-one (46):
A mixture of 45 (5.3 g, 18.9 mmol), 10% w/w Pd/C
(320 mg), ethyl acetate (30 mL), and CH2Cl2 (10 mL) was
hydrogenated in a Parr hydrogenation apparatus for 3 h
at 45 psi. The mixture was filtered through Celite and
concentrated, and the residue was chromatographed on
39
2138181
silica gel (ethyl acetate) to afford 46 (4.3 g, 80%).
3C NMR (CDCl3) ~ 177.64 (C), 83.87 (CH), 77.21 (CH),
76.02 (CH), 53.45 (CH), 43.84 (CH), 42.96 (CH), 40.46
(CH2), 35.88 (CH2), 31.37 (CH2), 29.23 (CH2), 29.10
(CH2), 27.85 (CH2), 26.41 (CH2), 26.22 (CH2), 26.08
( CH2 ) .
D: (3aR, 4R, 5R, 6aS)-4-~(3S)-3-Cyclohexyl-3-
(tetrahydroPYran-2-yloxy)propyl~-hexahydro-5-
(tetrahydropyran-2-yloxy)-2H-cyclopenta~b~furan-2-one
(47):
To a mixture of 46 (3.7 g, 12.9 mmol), DHP (2.9 g,
35 mmol), and CH2Cl2 (80 mL) at 0C was added pTSA (300
mg, 1.58 mmol). After 30 min 50 mL of saturated NaHC03
was added, the layers were separated, the aqueous layer
was extracted with CH2CL2 (2 X 50 mL), the combined
organic layers were dried over MgS04, filtered, and
concentrated, and the residue was chromatographed on
silica gel (1/1 hexane/ethyl acetate) to afford bis-THP
ether 47 (4.89 g, 89%).
E: (9S, llR, 15S)-11,15-Bis(tetrahYdropyran-2-yloxy)-
15-cyclohexyl-2,3,4,5,6,16,17,18,19,20-decanor-9-
hYdroxYprostanol (48):
To a suspension of lithium aluminum hydride (920
mg, 24.2 mmol) in 50 mL of THF at 0C was added
dropwise a solution of lactone 47 (10.87 g, 24.1 mmol)
in THF (100 mL). After stirring for 1 h, ethyl acetate
was cautiously added dropwise (50 mL), followed by
saturated NH4Cl (100 mL). The mixture was extracted
with ethyl acetate (3 X 100 mL), the combined organic
layers were dried over MgSO4, filtered, and
concentrated to provide crude diol 48 (9.73 g, 89%),
which was used in the next step without purification.
2138181
F: (9S, llR 15S)-11,15-Bis(tetrahydropyran-2-yloxy)-
15-cyclohexyl- 2 3 4,5 6 16 17 18 19 20-decanor-9-
(triethylsiloxy)prostanol triethylsilyl ether (49):
A mixture of 48 (9.7 g, 21.3 mmol), triethylsilyl
chloride (7.36 g, 48.9 mmol), DMAP (289 mg, 2.36 mmol),
NEt3 (5.3 g, 52 mmol), and CH2Cl2 (100 mL) was stirred
overnight at room temperature. The mixture was poured
into 100 mL of saturated NH4Cl, the layers were
separated, the aqueous layer was extracted with CH2Cl2
(50 mL), the combined organic layers were dried over
MgS04, filtered, and concentrated, and the residue was
chromatographed on silica gel (15% ethyl acetate in
hexane) to afford 49 (11.75 g, 81%).
G: (9S llR 15S)-11 15-Bis(tetrahydropyran-2-yloxy)-
15-cyclohexyl- 2 3 4 5,6 16 17 18 19 20-decanor-9-
(triethylsiloxy)prostanal (50):
To a solution of oxalyl chloride (6.5 g, 51 mmol)
in 30 mL of CH2Cl2 at -78C was added dropwise a
solution of DMSO (6.7 g, 86 mmol) in 10 mL of CH2Cl2.
After 30 min, a solution of 49 (11.75 g, 17.2 mmol) in
90 mL of CH2Cl2 was added dropwise. After 6 h, NEt3
(18.9 g, 187 mmol) was added and the reaction was
warmed to room temperature. The mixture was added to
150 mL of saturated NH4Cl, the layers were separated,
the aqueous layer was extracted with CH2Cl2 (2 X 50 mL),
the combined organic layers were dried over MgSO4,
filtered, and concentrated, and the residue was
chromatographed on silica gel (10% ethyl acetate in
hexane) to afford aldehyde 50 (8.75 g, 90%).
30 H: (5Z)-(9S, llR, 15S)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl- 2 3 4 16 17 18 19 20-octanor-9-
(triethylsiloxy)-5-prostenoic acid methyl ester (51):
To a mixture of 18-crown-6 (13.1 g, 49.6 mmol),
bis(2,2,2-trifluoroethyl)
(methoxycarbonylmethyl)phosphonate (7.8 g, 24.6 mmol),
~1381~1
and 75 mL of THF at -78C was added dropwise a 0.5 M
solution of KHMDS in toluene (42 mL, 21.5 mmol).
After 30 min, a solution of 54 (8.75 g, 15.4 mmol) in
THF (75 mL) was added dropwise. After 2.5 h methanol
was added (10 mL), the reaction was warmed to room
temperature and added to saturated NH4Cl (100 mL), the
layers were separated, the aqueous layer was extracted
with ethyl acetate (2 X 50 mL), the combined organic
layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel (15% ethyl acetate in hexane) to afford cis-
crotonate 51 (4.05 g, 44%), as well as a mixture of
cis:trans olefin isomers (2.3 g, 25%).
I: (5Z)-(95,llR, 15S)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl- 2,3,4,16,17,18,19,20-octanor-9-
(triethylsiloxy)-5-prostenol (52):
To a solution of 51 (4.0 g, 6.4 mmol) in 25 mL of
THF at 0C was added dropwise a 1.5 M solution of
DIBAL-H in toluene (10 mL, 15 mmol). The reaction was
brought to room temperature and stirred overnight. The
mixture was added to 75 mL of saturated sodium
potassium tartrate and stirred for 30 min, the layers
were separated, the aqueous layer was extracted with
ethyl acetate (2 X 50 mL), the combined organic layers
were dried over MgS04, filtered, and concentrated, and
the residue was chromatographed on silica gel (20%
ethyl acetate in hexane) to provide allyl alcohol 52
(2.84 g, 75%).
J: (5Z)-(9S llR 155)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-9-hydroxy- 2,3,4,16,17,18,19,20-
octanor-5-prostenol (53):
To a solution of 52 (1.7 g, 2.9 mmol) in 20 mL of
THF at 0C was added a 1 M solution of TBAF in THF (4.6
mL, 4.6 mmol). After 30 min 30 mL of saturated NH4Cl
was added, the mixture was extracted with ethyl acetate
42
2138181
(3 X 30 mL), the combined organic layers were dried
over MgSO4, filtered, and concentrated, and the residue
was chromatographed on silica gel (3/2 ethyl
acetate/hexane) to yield diol 53 (1.29 g, 93%).
K: (5Z)-(9S, llR, 15S)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-9-hydroxy-3-oxa-16,17,18,19,20-
pentanor-5-prostenoic acid t-butyl ester (54):
A mixture of 53 (1.29 g, 2.68 mmol), toluene (20
mL), 25% w/w aqueous NaOH (20 mL), BU4NHSO4 (175 mg,
0.52 mmol), and t-butyl bromoacetate (2.6 g, 13.5 mmol)
was vigorously stirred for 10 min at 0C and 45 min at
room temperature. The layers were separated, the
aqueous layer was extracted with ethyl acetate (2 X 20
mL), the combined organic layers were dried over MgSO4,
filtered, and concentrated, and the residue was
chromatographed on silica gel (40% ethyl acetate in
hexane) to afford 54 (1.56 g, 98%).
L: (5Z)-(9S, llR, 155)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-9-hydroxy-3- oxa-16,17,18,19,20-
pentanor-5-prostenoic acid (55):
A mixture of 54 (2.0 g, 3.4 mmol), lithium
hydroxide monohydrate (830 mg, 20 mmol), methanol (20
mL), and water (1 mL) was stirred for 1.5 h, 35 mL of
saturated KH2PO4 was added to adjust the pH to ca. 6,
and the mixture was extracted with ethyl acetate (3 X
25 mL). The combined organic layers were dried over
Na2SO4, filtered, and concentrated to provide crude acid
55, which was used in the next step without
purification.
M: (5Z)-(9S, llR, 155)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-9-hydroxy-3- oxa-16,17,18,19,20-
pentanor-5-prostenoic acid isopropyl ester (56):
A mixture of 55 from above, acetone (34 mL),
isopropyl iodide (2.9 g, 17 mmol), and DBU (3.0 g, 20
2138181
mmol) was stirred overnight. The reaction was added to
40 mL of saturated NH4Cl, the mixture was extracted
with ethyl acetate (3 X 25 mL), the combined organic
layers were dried over MgS04, filtered, and
concentrated, and the residue was chromatographed on
silica gel (1/1 ethyl acetate/hexane) to yield
isopropyl ester 56 (1.26 g, 64% from t-butyl ester 58).
N: (5Z)-(9R, llR, 15S)-11,15-Bis(tetrahydropYran-2-
yloxy)-9-chloro-15-cyclohexYl-3- oxa-16,17,18,19,20-
pentanor-5-prostenoic acid isopropyl ester (57):
To a solution of 56 (1.26 g, 2.2 mmol) in 13 mL of
pyridine at 0C was added dropwise methanesulfonyl
chloride (600 mg, 5.2 mmol). After 2 h, the mixture
was added to a suspension of BU4NCl (6.1 g, 21.9 mmol)
in toluene (9 mL) and heated at 55-63C for 4 h. After
the reaction cooled to room temperature, 30 mL of
saturated NH4Cl was added, the mixture was extracted
with ethyl acetate (3 X 30 mL), the combined organic
layers were dried over MgS04, filtered, and
concentrated, and the residue was chromatographed on
silica gel (20% ethyl acetate in hexane) to provide
chlorinated product 57 as a mixture with the
corresponding 8,9-olefin (1.07 g, 81% calculated as the
chloride). The 8,9-olefin by-product was separated in
the next step.
0: (5Z)-(9R, llR, 15S)-9-Chloro-15-cYclohexyl-11,15-
dihydroxy-16,17,18,19,20-pentanor-5-prostenoic acid
isopropyl ester (XIV):
The mixture from above containing 57 and the 8,9-
olefin (1.07 g, 1.79 mmol, calculated as the chloride)was added to a solution of isopropanol (14 mL), water
(1 mL), and 12 M HCl (2 mL), and was stirred for 2 h.
The reaction was added to 50 mL of saturated NaHC03,
the mixture was extracted with ethyl acetate (3 X 30
mL), the combined organic layers were dried over MgS04,
44
2138181
filtered, and concentrated, and the residue was
purified by radial chromatography (5% isopropanol in
toluene) to afford pure XIV (172 mg, 22%), as well as a
mixture of containing mostly XIV and a smaller
proportion (ca. 30%) of the corresponding 8,9-olefin
(333 mg, 42% calculated as the chloride). 13C NMR
(CDCl3) o~ 170.21 (C), 131.12 (CH), 127.19 (CH), 76.75
(CH), 75.44 (CH), 68.70 (CH), 67.67 (CH2), 66.78 (CH2),
61.02 (CH), 54.28 (CH), 51.18 (CH), 44.31 (CH2), 44.19
10 (CH), 31.35 (CH2), 31.19 (CH2), 29.73 (CH2), 29.17 (CH2),
27.60 (CH2) 26.50 (CH2), 26.31 (CH2), 26.16 (CH2), 21.80
(CH3). CI MS, m/z calcd. for C23H~O5Cl (MH+) 431, found
431.
EXA~P~E 11S 8YNTHE8I8 OF CONrOu~v XV
v s~ s~
C~ a~ C~_o~co2a~-
~0--~0 ~0
.. .. ..
.~o~c~
HO~Q
~V
A: (SZ)-(9R, llR, 15R)-9-Chloro-15-cyclohexyl-11,15-
dihydroxy-3-oxa-16,17 18,19,20-~entanor-S-prostenoic
acid (58):
To a solution of 32 mg (0.072 mmol) of V (Example
1) in a mixture of 2.5 mL of methanol and 1.2 mL of
water was added lS mg (0.36 mmol) of lithium hydroxide
monohydrate. After 9 h, 5 mL of 0.1 M HCl was added,
and the mixture was extracted with CHC13 (S X 5 mL).
The combined organic layers were washed with water (20
mL) and brine (20 mL), dried over Na2SO4, filtered, and
2138181
concentrated to afford 30 mg (100%) of 58 as a
colorless oil. 13C NMR (CDCl3) ~ 173.17 (C), 132.77
(CH), 126.03 (CH), 75.68 (CH), 75.24 (CH), 66.89 (CH2),
66.40 (CH2), 61.32 (CH), 54.12 (CH), 50.62 (CH), 43.59
(CH2), 43.33 (CH), 30.92 (CH2), 30.73 (CH2), 29.89
(CH2), 29.30 (CH2), 27.95 (CH2), 26.44 (CH2), 26.25
(CH2), 26.09 (CH2).
B: (5Z~-(9R, llR, 15R)-9-Chloro-15-cYclohexy~ l5-
dihydroxy-3-oxa-16 17 18 19 20-pentanor-5-~rostenoic
acid methyl ester (59):
To a solution of 58 (22 mg, 0.057 mmol) in 20 mL
of ether was added excess diazomethane as a solution in
ether. After 20 min residual diazomethane was removed
from solution by a stream of N2 and the solution was
concentrated. The residue was chromatographed on
silica gel (40% ethyl acetate in hexane) to afford 59
(21 mg, 91%). 13C NMR (CDCl3) ~ 171.06 (C), 131.75
(CH), 126.74 (CH), 75.73 (CH), 75.47 (CH), 67.34 (CH2),
66.77 (CH2), 61.08 (CH), 54.19 (CH), 51.94 (CH3), 51.29
(CH), 44.47 (CH2), 43.62 (CH), 31.49 (CH2), 30.03 (CH2),
29.30 (CH2), 27.98 (CH2), 26.49 (CH2), 26.28 (CH2), 26.13
(CH2) .
C: (5Z)-(9R llR, 15R)-11 15-Bis(tetrahydropyran-2-
yloxy)-9-chloro-15-cyclohexyl-3-oxa-16,17 18 19,20-
pentanor-5-prostenoic acid methyl ester (60):
To a mixture of diol 59 (645 mg, 0.65 mmol), DHP
(0.56 mL, 6.4 mmol), and CH2Cl2 (10 mL) at 0C was added
pTSA (10 mg, 0.053 mmol). After 15 min, 15 mL of
saturated NaHC03 was added, the layers were separated,
the aqueous layer was extracted with CH2Cl2 (3 X 15 mL),
the combined organic layers were dried over MgS04,
filtered, and concentrated, and the residue was
chromatographed on silica gel (30% ethyl acetate in
hexane) to afford 781 mg (86%) of bis-THP ether 60.
46
2138181
.
D: (5Z)-(9R, llR, 15R)-11,15-Bis(tetrahydropyran-2-
yloxy)-9-chloro-15-cyclohexyl-3-oxa-16,17,18,19,20-
pentanor-5-prostenoic acid (61):
A solution of 60 (342 mg, 0.65 mmol) and lithium
hydroxide monohydrate (96 mg, 2.3 mmol) in methanol (20
mL) was stirred for 2 h, the pH of the solution was
adjusted to pH 6 with saturated KH2P04, and the mixture
was extracted with CH2Cl2 (6 X 20 mL). The combined
organic layers were dried over Na2S04, filtered, and
concentrated to afford acid 61 (334 mg, 92%).
E: (5Z)-(9R, llR, 15R)-11,15-Bis(tetrahydropyran-2-
yloxY)-9-chloro-15-cYclohexyl-3-oxa-16 17,18,19 20-
pentanor-5-prostenoic acid neopentyl ester (62):
A mixture of 61 (80 mg, 0.14 mmol),
dicyclohexylcarbodiimide (30 mg, 0.18 mmol), DMAP (8
mg, 0.07 mmol), 2,2-dimethyl-1-propanol (60 mg, 0.7
mmol), and CH2Cl2 (1 mL) was stirred for 3 h at room
temperature, the solution was concentrated, and the
residue was chromatographed on silica gel (20% ethyl
acetate in hexane) to provide 62 contaminated with some
dicyclohexyl urea (110 mg total). The sample was used
in the next reaction without further purification.
F: (5Z)-(9R, llR, 15R)-9-Chloro-15-cyclohexyl-11,15-
dihYdroxy-3-oxa-16,17,18,19,20-pentanor-5-Prostenoic
acid neopentyl ester (XV):
The llo mg sample from above was dissolved in a
mixture of methanol (10 mL) and water (1 mL) at 0C,
and 10 drops of 12 N HCl was added. After 15 min, the
solution was warmed to room temperature for 30 min.
The solution was added to CH2Cl2 (10 mL), the layers
were separated, and the aqueous layer was extracted
with CH2Cl2 (3 X 10 mL). The combined organic layers
were dried over MgSO4, filtered, concentrated, and
chromatographed on silica gel (1/1 ethyl
acetate/hexane) to afford XV (46 mg, 71% from 61).
47
2133181
3C NMR (CDCl3) ~ 170.88 (C), 131.82 (CH), 126.72 (CH),
75.68 (CH), 75.23 (CH), 74.20 (CH2), 67.34 (CH2) 66.76
(CH2), 61.23 (CH), 54.24 (CH), 51.19 (CH), 44.43 (CH2),
43.65 (CH), 31.44 (CH2), 31.34 (C), 30.23 (CH2), 30.09
5 (CH2), 29.33 (CH2), 28.00 (CH2), 26.50 (CH2), 26.37
(CH3), 26.28 (CH2), 26.14 (CH2) . CI MS m/z calcd. for
C25H~OsCl (MH+) 459.2877, found 459.2872.
48
21381~1
EXA~IPLE: 12 s ~ I8 OF COMPOUND ~
.f~ H t .~ o^co~ o^~l,OH
~0 O ' ' ~0 ''
32 o ~
~3 ~
A: (5Z. 13E)-(9S, llR, 15S)-11,15-Bis(tetrahydropYran-
2-Yloxy)-15-cyclohexyl-9-hydroxY-3-oxa-16,17,18,19,20-
pentanor-5,13-prostadienoic acid t-butYl ester (63)
To a vigorously stirring mixture of diol 32 (see
Example 6) (925 mg, 1.93 mmol), toluene (25 mL),
BrCH2CO2Bu' (1.2 g, 6.3 mmol), and Bu4HSO4 (200 mg, 0.59
mmol) was added 20 mL of a 20% w/w aqueous solution of
NaOH. After 40 min the layers were separated, the
aqueous layer was extracted with ethyl acetate (2 X 30
mL), the combined organic layers were dried over MgSO4,
filtered, and concentrated, and the residue was
chromatographed on silica gel eluting with 40% ethyl
acetate in hexanes to afford 63 (1.10 g, 96%, ~0.5).
B: (5Z 13E)-(llR, 15S)-11,15-Bis(tetrahydropyran-2-
YloxY)-15-cyclohexYl-3-oxa-16,17 18,19 20-~entanor-
5,13-prostadienol (64)
To a mixture of 63 (1.09 g, 1.84 mmol) and
pyridine (11 mL) at 0C was added dropwise ~
methanesulfonyl chloride (0.51 g, 4.5 mmol). The
reaction was stirred at 0C~for 30 min and for 2 h at
room temperature, 40 mL of saturated NH4Cl was added,
and the mixture was extracted with ethyl acetate (2 X
40 mL). The combined organic layers were dried over
49
213~181
MgSO4, filtered, and concentrated, and the residue was
chromatographed on silica gel eluting with 1:1
hexane:ethyl acetate to afford the intermediate 9~-
mesylate (1.08 g, 87%, RfO.5).
To a mixture of the mesylate in THF (11 mL) at 0C
was added dropwise 1 M LiEt3BH in THF (11 mL, 11 mmol)
and the reaction was stirred overnight at room
temperature. The mixture was poured into 50 mL of a
1:1 mixture of saturated NH4Cl:ethyl acetate, the
layers were separated, and the aqueous layer was
extracted with ethyl acetate (2 X 30 mL). The combined
organic layers were dried over MgSO4, filtered, and
concentrated, and the residue was chromatographed on
silica gel eluting with 40% ethyl acetate in hexane to
afford 64 (642 mg, 79%, RfO.4).
C: (5Z, 13E)-(llR, 15S)-11,15-Bis(tetrahydropyran-2-
yloxy)-15-cyclohexyl-3-oxa-16,17,18,19,20-pentanor-
5,13-prostadienoic acid isopropyl ester (65)
To a mixture of 64 (560 mg, 1.12 mmol) and DMF (5
mL) was added pyridinium dichromate (1.32 g, 3.51
mmol). After stirring for 48 h, 20 mL of water was
added, and the mixture was extracted with ethyl acetate
(4 X 20 mL). The combined organic layers were filtered
and concentrated, the residue was dissolved in acetone
(20 mL), and DBU was added (780 mg, 5.12 mmol),
followed in 15 min by the addition of isopropyl iodide
(850 mg, 5.0 mmol). After 20 h, the reaction was
concentrated, and the residue was chromatographed on
silica gel eluting with 20% ethyl acetate in hexane to
afford 65 (238 mg, 38%, RfO.4).
D: (5Z, 13E)-(llR, 15S)-15-Cyclohexyl-11,15-dihydroxy-
3-oxa-16,17,18,19,20-pentanor-5,13-prostadienoic acid
isopropyl ester (XVI)
To a mixture of 65 (230 mg, 0.41 mmol),
isopropanol (18 mL), and water (2 mL) was added 12 M
2 1 ~
HCl (1 mL). After 2 h, 20 mL of saturated NaHCO3 was
added, the mixture was extracted with ethyl acetate (3
X 20 mL), the combined organic layers were dried over
MgSO4, filtered, and concentrated, and the residue was
chromatographed on silica gel eluting with 3:2 ethyl
acetate:hexane to afford XVI (120 mg, 74%, ~0.25).
3C NMR (CDCl3) ~ 170.00 (C), 134.17 (CH), 132.73 (CH),
125.97 (CH), 77.92 (CH), 77.68 (CH), 68.47 (CH), 67.38
(CH2), 66.73 (CH2), 58.02 (CH), 43.44 (CH), 42.77 (CH),
32.15 (CH2), 28.89 (CH2), 28.80 (CH2), 27.63 (CH2), 26.51
(CH2), 26.06 (CH2), 25.94 (CH2), 21.97 (CH3).
The compounds of formulae (I) and (II) are useful
in lowering intraocular pressure and thus are useful in
the treatment of glaucoma. The preferred route of
administration is topical. The dosage range for
topical administration is generally between about 0.001
and about 1000 micrograms per eye (~g/eye) and is
preferably between about 0.01 and about 100 ~g/eye and
most preferably between about 0.05 and 50 ~g/eye. The
compounds of the present invention can be administered
as solutions, suspensions, or emulsions (dispersions)
in a suitable ophthalmic vehicle.
In forming compositions for topical
administration, the compounds of the present invention
are generally formulated as between about 0.00002 to
about 0.5 percent by weight (wt%) solutions in water at
a pH between about 4.5 and about 8Ø The compounds
are preferably formulated as between about 0.0001 to
about 0.1 wt% and, most preferably, between about 0.001
and about 0.05 wt%. While the precise regimen is left
to the discretion of the clinician, it is recommended
that the compositions be topically applied by placing
one or more drops in each eye one or more times a day.
Other ingredients which may be desirable to use in
the ophthalmic preparations of the present invention
~138181
include preservatives, co-solvents and viscosity
building agents.
Antimicrobial Preservatives:
Ophthalmic products are typically packaged in
multidose form, which generally require the addition of
preservatives to prevent microbial contamination during
use. Suitable preseryatives include: benzalkonium
chloride, thimerosal, chlorobutanol, methyl paraben,
propyl paraben, phenylethyl alcohol, edetate disodium,
sorbic acid, ONAMER M~, or other agents known to those
skilled in the art. Such preservatives are typically
employed at a concentration between about 0.001 and
about 1.0 wt%.
Co-Solvents:
Prostaglandins, and particularly ester
derivatives, typically have limited solubility in water
and therefore may require a surfactant or other
appropriate co-solvent in the composition. Such co-
solvents include: Polysorbate 20, 60 and 80; Pluronic
F-68, F-84 and P-103; Tyloxapol; Cremophor EL; sodium
dodecyl sulfate; glycerol; PEG 400; propylene glycol;
cyclodextrins; or other agents known to those skilled
in the art. Such co-solvents are typically employed at
a concentration between about 0.01 and about 2 wt%.
Viscosity Agents:
Viscosity greater than that of simple aqueous
solutions may be desirable to increase ocular
absorption of the active compound, to decrease
variability in dispensing the formulations, to decrease
physical separation of components of a suspension or
emulsion of formulation and/or otherwise to improve the
ophthalmic formulation. Such viscosity building agents
include: polyvinyl alcohol; polyvinyl pyrrolidone;
cellulosic polymers, such as methyl cellulose, hydroxy
propyl methylcellulose, hydroxyethyl cellulose,
2138181
carboxymethyl cellulose, hydroxy propyl cellulose;
carboxy vinyl polymers, such as carbomer 910, carbomer
940, carbomer 934P and carbomer 1342; or other agents
known to those skilled in the art. Such agents are
typically used at a concentration between about 0.01
and about 2 wt~.
53
213Sl~l
EXAMPLE 13
The following Tables 2-4 containing Formulations
A-M are representative pharmaceutical compositions of
the invention for topical use in lowering of
intraocular pressure. Each of Formulations A-M may be
formulated in accordance with procedures known to those
skilled in the art.
TABLE 2
FORMULATION (wt%)
INGREDIENT A B C D
Compound lll --- 0.01 --- ---
Compound IV 0.002 --- --- ---
Compound XVI --- --- 0 05 ---
Compound XVII --- --- --- 0.1
Dextran 70 0.1 0.1 -- ---
Hydroxypropyl 0.3 0.5 --- ---
Methylcellulose
Monobasic Sodium --- 0.05 0.05 0.05
Phosphate
Dibasic Sodium --- 0.15 0.15 0.15
Phosphate (anhydrous)
Sodium Chloride 0.77 0.75 0.5 0.6
Potassium Chloride0.12 --- --- ---
Disodium EDTA 0.05 0.05 --- ---
Hydroxypropyl-,~- --- --- 1.0 ---
cyclodextrin
Tyloxapol --- 0.1 --- 0.4
Benzalkonium Chloride 0.01 0.01 0.01 0.02
Polysorbate 80 0.01 --- --- ---
HCI and/or NaOHq.s. to pHq.s. to pHq.s. to pHq.s. to pH
7.2-7.5 7.3-7.4 6.3-6.6 6.3-6.6
Purified Water q.s. to q.s. to q.s. to q.s. to
100% 100% 100% 100%
54
2138181
TABLE 3
FORMULATION (wt%)
INGREDIENT E F G H
Compound V 0.01 --- --- ---
Compound Vl --- 0.01 --- ---
Compound XVII --- --- 0.1 ---
Compound XIX --- --- --- 0.2
Monobasic Sodium 0.05 0.05 0.05 0.05
Phosphate
Dibasic Sodium 0.15 0.15 0.15 0.15
Phosphate (anhydrous)
Sodium Chloride 0.75 0.75 0.5 0.6
Disodium EDTA 0.01 0.05 --- ---
Cremophor~ EL --- 01 --- ---
Hydroxypropyl-,~- --- , --- 4.0 ---
cyclodextrin
Tyloxapol --- --- --- 0.5
Benzalkonium Chloride 0.02 0.01 0.01 0.02
Polysorbate 80 0.05 --- --- ~~~
HCI and/or NaOHq.s. to pHq.s. to pHq.s. to pHq.s. to pH
7.3-7.4 7.3-7.4 6.3-6.6 6.3-6.6
Purified Water q.s. to q.s. to q.s. to q.s. to
100% 100/0 100% 100%
21~8181
TABLE 4
FORMULATION (wt%)
INGREDIENT J K L M
Compound Vlll 0.01 --- --- ---
Compound IX --- 0.01 --- ---
Compound X --- --- 0.1 0.2
Monobasic Sodium 0.05 0.05 0.05 0.05
Phosphate
Dibasic Sodium 0.1 5 0.1 5 0.1 5 0.1 5
Phosphate (anhydrous)
Sodium Chloride 0.75 0.75 0.5 0.6
Disodium EDTA 0.01 0.05 --- ---
Cremophor~ EL --- 0 01 --- ---
Hydroxypropyl-,~- --- --- 4.0 ---
cyclodextrin
Tyloxapol --- --- --- 0.5
Benzalkonium Chloride 0.02 0.01 0.01 0.02
Polysorbate 80 0.05 --- --- ~~~
HCI and/or NaOHq.s. to pHq.s. to pHq.s. to pHq.s. to pH
7.3-7.4 7.3-7.4 6.3-6.6 6.3-6.6
Purified Water q.s. to q.s. to q.s. to q.s. to
100% 100% 100% 100%
56
213~
EXAMPLE 14
The ability of certain compounds of the present
invention to reduce intraocular pressure (IOP) was
evaluated in cynomolgus monkeys with ocular
hypertension produced by previous laser trabeculoplasty
in the right eye. Animals had been trained to sit in
restraint chairs and conditioned to accept experimental
procedures without chemical restraint. IOP was
determined with a pneumatonometer after light corneal
anesthesia with dilute proparacaine. The test protocol
included a treatment regimen consisting of 5 divided
doses administered over a period of 2 and 1/2 days.
Doses Z-5 were given 8, 24, 32 and 48 hours after the
initial dose. Baseline IOP values were determined
prior to treatment with the test formulation, and then
IOP was determined 16 hours after the fourth dose, and
2, 4 and 6 hours after the fifth dose. Prostaglandin
doses are micrograms of compound contained in each
treatment.
The two tested compounds are the previously
identified Compounds III-VII.
57
TABLE 5
Baseline Percent IOP Reduction at Hours after Dose/Dose~
Compound PG Dose IOP
(mm Hg) 16/4 215 415 615
111 1/J9 31.1 15.6+4.733.7+5.5 28.1 +4.131.0~4.6
IV 1,ug 31.3 47.6+2.652.8+3.3 54.7+3.8 53.1 +4.1
V 5 ~9 36.3 38.3+3.047.5+5.1 43.7+5.6 36.5+5.8
Vl 1 IJ9 36.9 22.3+2.730.8+4.7 26.4+4.7 24.9+4.0
Vll 20,ug 32.6 13.9+2.730.1 i4.822.4+5.2 19.3~4.3 2
C~
58
213~
Results are presented in Table S, above.
Compounds III-VII produced significant reduction of
intraocular pressure at doses which are marginal or
ineffective for other prostaglandins in published
clinical studies. Compound IV was especially potent,
producing greater than 50% reduction of intraocular
pressure with just 1 ~g of compound. In contrast,
Nakajima et al. (Graefe's Arch. Clin. Exp. Ophthalmol.,
229:411-413 (1991)) reported that 50 ~g of PGD2 and 2.5
~g of BW245C (a PGD2 analogue) reduced intraocular
pressure in human eyes by 12% and 10%, respectively.
Other studies (Woodward et al., Invest. Ophthalmol.
Vis. Sci., 31:138-146 (1990)) reported for these
reference compounds in rabbits describe a maximum IOP
reduction of approximately 28% for 250 ~g of PGD2 and
22% for 25 ~g of BW245C. These comparisons indicate
the unexpected potency of Compounds III - VII in
reducing intraocular pressure. No indications of
inflammation were observed during these studies.
EXAMPLE 15
The ability of certain compounds of the present
invention to reduce intraocular pressure (IOP) was
evaluated in cynomolgus monkeys with ocular
hypertension produced by previous laser trabeculoplasty
in the right eye. Animals had been trained to sit in
restraint chairs and conditioned to accept experimental
procedures without chemical restraint. IOP was
determined with a pneumatonometer after light corneal
anesthesia with dilute proparacaine.
The two compounds tested are those previously
identified as Compound XI and Compound XII.
2l38I8l
TABLE 7
MAXIMUM PERCENT IOP
REDUCTION FROM
COMPOUND PG DOSE BASELINE
Xl 3,ug 42
Xll 0.3,ug 44
Compounds XI and XII produced significant
reduction of intraocular pressure at doses which are
marginal or ineffective for other prostaglandins in
published clinical studies. By comparison, Nakajima et
al. (Graefe's Arch. Clin. Exp. OPhthalmol. 229:411-413
(1991)) reported that 50 ~g of PGD2 and 2.5 ~g of
BW245C (a PGD2 analogue) reduce intraocular pressure in
human eyes by 12% and 10%, respectively. Other studies
(Woodward et al., Invest. Ophthalmol. Vis. Sci. 31:138-
146 (1990)) reported for these reference compounds in
rabbits describe a maximum IOP reduction of
approximately 28% for 250 ~g of PGD2 and 22% for 25 ~g
of BW245C. These comparisons indicate the unexpected
potency of compounds of the present invention in
reducing intraocular pressure. No indications of
inflammation were observed during these studies.
The invention has been described by reference to
certain preferred embodiments; however, it should be
understood that it may be embodied in other specific
forms or variations thereof without departing from its
spirit or essential characteristics. The embodiments
described above are therefore considered to be
illustrative in all respects and not restrictive, the
scope of the invention being indicated by the appended
claims rather than by the foregoing description.
