PCT
`
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(51) International Patent Classification 6 :
`C07C 405/00
`
`(11) International Publication Number:
`
`WO 98/21181
`
`(43) International Publication Date:
`
`22 May 1998 (22.05.98)
`
`(21) International Application Number:
`
`PCT/US97/20671
`
`(22) International Filing Date:
`
`7 November 1997 (07.11.97)
`
`(81) Designated States: AU, CA, CN, JP, KR, MX, US, European
`patent (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT,
`LU, MC, NL, PT, SE).
`
`(30) Priority Data:
`60/030,519
`
`12 November 1996 (12.11.96)
`
`US
`
`Published
`
`Without international search report and to be republished
`upon receipt of that report.
`
`(71) Applicant (for all designated States except US): ALCON
`LABORATORIES, INC.
`[US/US]; 6201 South Freeway,
`Fort Worth, TX 76134—2099 (US).
`
`(72) Inventors; and
`(75) Inventors/Applicants (for US only): KLIMKO, Peter, G.
`[US/US]; 2115 Pembroke Drive, Fort Worth, TX 76110
`(US). HELLBERG, Mark, R.
`[US/US]; 5211 Overridge
`Drive, Arlington, TX 76017 (US). ZlNKE, Paul, W.
`[US/US]; 4129 Willow Way Road, Fort Worth, TX 76133
`(US).
`
`
`
`(74) Agents: COPELAND, Barry, L. et al.; Alcon Laboratories, Inc.,
`Patent Dept, Q—148, 6201 South Freeway, Fort Worth, TX
`76134—2099 (US).
`
`(54) Title:
`
`15—FLUORO PROSTAGLANDINS AS OCULAR HYPOTENSIVES
`
`(57) Abstract
`
`IS—fluoro substituted analogs of PGan and methods of their use in treating glaucoma and ocular hypertension are disclosed.
`
`Micro Labs Exhibit 1012
`
`Micro Labs Exhibit 1012
`
`

`

`
`
`Albania
`Armenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`C6te d’Ivoire
`Cameroon
`China
`Cuba
`Czech Republic
`Germany
`Denmark
`Estonia
`
`SI
`SK
`SN
`SZ
`TD
`TG
`TJ
`TM
`TR
`TT
`UA
`UG
`US
`UZ
`VN
`YU
`ZW
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`Uganda
`United States of America
`Uzbekistan
`Viet Nam
`Yugoslavia
`Zimbabwe
`
`
`
`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
`
`ES
`Fl
`FR
`GA
`GB
`GE
`GI-I
`GN
`GR
`HU
`IE
`IL
`IS
`IT
`JP
`KE
`KG
`KP
`
`KR
`KZ
`LC
`LI
`LK
`LR
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Ireland
`Israel
`Iceland
`Italy
`Japan
`Kenya
`Kyrgyzstan
`Democratic People's
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`R0
`RU
`SD
`SE
`SG
`
`Lesotho
`Lithuania
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The former Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mexico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`Micro Labs Exhibit 1012-2
`
`Micro Labs Exhibit 1012-2
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`15-FLUORO PROSTAGLANDINS AS OCULAR HYPOTENSIVES
`
`Background of the Invention
`
`
`
`can affect patient compliance and/or necessitate the termination of treatment. Some beta—
`
`The present invention relates to compounds for the treatment of glaucoma and
`
`ocular hypertension. In particular, the present invention relates to the use of certain 15-
`
`fluoro analogs of F series prostaglandins to treat glaucoma and ocular hypertension.
`
`Glaucoma is a progressive disease which leads to optic nerve damage, and,
`
`ultimately, total loss of vision. The causes of this disease have been the subject of extensive
`
`studies for many years, but are still not fully understood. The principal symptom of and/or
`
`risk factor for the disease is elevated intraocular pressure or ocular hypertension due to
`
`excess aqueous humor in the anterior chamber of the eye.
`
`The causes of aqueous humor accumulation in the anterior chamber are not fully
`
`understood. It is known that elevated intraocular pressure (“IOP”) can be at least partially
`
`controlled by administering drugs which either reduce the production of aqueous humor
`
`within the eye, such as beta—blockers and carbonic anhydrase inhibitors, or increase the flow
`
`of aqueous humor out of the eye, such as miotics and sympathomimetics.
`
`Most types of drugs conventionally used to treat glaucoma have potentially serious
`
`side effects. Miotics such as pilocarpine can cause blurring of Vision and other visual side
`
`effects, which may lead either to decreased patient compliance or to termination of
`
`therapy. Systemically administered carbonic anhydrase inhibitors can also cause serious
`
`side effects, such as nausea, dyspepsia, fatigue, and metabolic acidosis, which side effects
`
`blockers have increasingly become associated with serious pulmonary side effects
`
`_1-
`
`Micro Labs Exhibit 1012-3
`
`Micro Labs Exhibit 1012-3
`
`

`

`WO 98/21181
`
`PCT/US97/2067l
`
`attributable to their effects on beta—2 receptors in pulmonary tissue. Sympathomimetics
`
`may cause tachycardia, arrhythmia and hypertension. There is therefore a continuing need
`
`for therapies which control the elevated intraocular pressure associated with glaucoma.
`
`Prostaglandins, which are metabolite derivatives of arachidonic acid, have recently
`
`been pursued for possible efficacy in lowering IOP. Arachidonic acid in the body is
`
`converted to prostaglandin G2, which is subsequently converted to prostaglandin H2.
`
`Other naturally occurring prostaglandins are derivatives of prostaglandin H2. A number of
`
`different types of prostaglandins have been discovered including A, B, D, E, F, G, I and J-
`
`Series prostaglandins (EP 0 561 073 A1). Of interest in the present invention are
`
`compounds which are believed to exhibit IOP lowering effects similar to those exhibited
`
`by PGFZa (an F-series prostaglandin):
`
`
`
`aqueous humor. Regardless of mechanism, PGFZQ and analogs have been shown to lower
`
`The relationship of PGF20L receptor activation and IOP lowering effects is not well
`
`understood. It is believed that PGFZQ receptor activation leads to increased outflow of
`
`IOP (Giuffre, The Eflects ofProstaglandin F2,, the Human Eye, giraefe’s Archive
`
`Ophthalmology, volume 222, pages 139-141 (1985); and Kerstetter et al., Prostaglandz'n
`
`F26,—] -Is0propylester Lowers Intraocular Pressure Without Decreasing Aqueous Humor
`
`Flow, American Journal of Ophthalmology, volume 105, pages 30-34 (1988)). Thus, it
`
`has been of interest in the field to develop synthetic PGFZ0c analogs with IOP lowering
`
`efficacy.
`
`Micro Labs Exhibit 1012-4
`
`Micro Labs Exhibit 1012-4
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`Synthetic PGFZa-type analogs have been pursued in the art (g iraefe’s Archive
`
`g ththalmology, volume 229, pages 411—413 (1991)). Though PGFZa-type molecules
`
`lower IOP, many of these types of molecules have also been associated with undesirable
`
`side effects resulting from topical ophthalmic dosing. Such effects include an initial
`
`increase in IOP, breakdown of the blood aqueous barrier and conjunctival hyperemia
`
`inion
`
`in Qphthalmology, volume 4, No. 11, pages 44-50 (1993)).
`
`Based on the foregoing, a need exists for the development of molecules that may
`
`activate PGFZQ receptors, yielding a more efficacious lowering of IOP, while exhibiting
`
`fewer or reduced side effects.
`
`An agent which exhibits the same or improved efficacy, but with reduced side
`
`effects when compared to other agents, is said to have an improved therapeutic profile. It
`
`is an obj ect of this invention to provide a class of IOP lowering agents with an improved
`
`therapeutic profile over PGFZQ, and methods of their use. It has now unexpectedly been
`
`discovered that the presently claimed 15-fluoro analogs of PGFZOL meet this objective.
`
`While some prostaglandins with fluorine in the omega chain are known in the art [EP
`
`435,443 A; JP 7,070,054 A2; Eksp. Klin. Farmakol., volume 57, number 2, pages 39-41
`
`(1994) (Chemical Abstracts, volume 121, abstract 50656 (1994)); Izv. Akad. Nauk SSSR,
`
`M, volume 6, pages 831—7 (1989) (W, volume 112, abstract 30749
`
`(1990))], the novel compounds of the present invention and their favorable therapeutic
`
`profiles in the treatment of glaucoma are neither disclosed nor suggested in that art.
`
`(Alm, The Potential ofProstaglandin Derivatives in Glaucoma Therapy, urrent
`
`
`
`Summary of the Invention
`
`The present invention is directed to compositions and methods of their use in
`
`treating IOP and ocular hypertension. In particular, the present invention provides 15-
`
`fluoro prostaglandin analogs believed to have functional PGF20L receptor agonist activity,
`
`-3-
`
`Micro Labs Exhibit 1012-5
`
`Micro Labs Exhibit 1012-5
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`and methods of their use in treating glaucoma and ocular hypertension. As previously
`
`stated, the mechanism of action by which PGFZO, type prostaglandins lower IOP is not well
`
`understood While the mechanism of action of the compounds of the present invention is
`
`not fully understood, the inventors theorize that such compounds exhibit enhanced FP
`
`receptor selectivity as a consequence of their decreased activity at the EP receptor site.
`
`While bound by no such theory, it is possible that an improved therapeutic index may
`
`result from a relative reduction of EP-mediated side—effects.
`
`Detailed Description of the Invention
`
`It has unexpectedly been found that 15-fluoro substituted PGF20c analogs of the
`
`present invention exhibit an improved therapeutic profile in the treatment of glaucoma and
`
`ocular hypertension when compared to natural prostaglandins and many of their known
`
`analogs. The substituted PGFZO, analogs of the present invention have the following
`
`formula I:
`
`
`
`H, acyl, or alkyl; with the proviso that if one of R7, R8 = acyl, then the other = H or
`
`wherein:
`
`R1 = co,R, C0NR4R5, CH20R6, or CHZNR7R8, where
`
`R = H or cationic salt moiety, or COZR = pharmaceutially acceptable ester moiety; R4,
`
`R5 = same or different = H or alkyl; R6 = H, acyl, or alkyl; R7, R8 = same or different =
`
`alkyl;
`
`Micro Labs Exhibit 1012-6
`
`Micro Labs Exhibit 1012-6
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`n=00r2;
`
`--—- = single or non—cumulated double bond, with the provisos that a double bond between
`
`carbons 4 and 5 may not be of the trans configuration; and that a double bond between
`
`carbons 13 and 14 may not be of the cis configuration;
`
`R2, R3 = same or different = H, alkyl, or acyl;
`
`D, D1 = different = H and fluorine;
`
`X = (CH2)q or (CH2)qO; where q = 1—6; and
`
`Y = a phenyl ring optionally substituted with alkyl, halo, trihalomethyl, alkoxy, acyl,
`
`acyloxy, amino, alkylamino, acylamino, or hydroxy; or
`
`X—Y = (CH2)pY1; where p = 0—6; and
`
`
`
`W = CH2, 0, S(O)m, NR9, CHZCHZ, CH=CH, CHZO, CHZS(O)m, CH=N, or
`
`wherein:
`
`CHZNRg; where m = 0—2, and R9 = H, alkyl, or acyl;
`
`Z = H, alkyl, alkoxy, acyl, acyloxy, halo, trihalomethyl, amino, alkylamino,
`
`acylamino, or hydroxy; and
`
`--—— = single or double bond.
`
`Micro Labs Exhibit 1012-7
`
`Micro Labs Exhibit 1012-7
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`For purposes of the foregoing definition, the term “pharmaceutically acceptable
`
`ester” means any ester that would be suitable for therapeutic administration to a patient by
`
`any conventional means without significant deleterious health consequences; and
`
`“ophthalmically acceptable ester” means any pharmaceutically acceptable ester that would
`
`be suitable for ophthalmic application, i.e. non—toxic and non—irritating. Preferred among
`
`the ophthalmically acceptable esters are alkyl esters. Most preferred are C2-C4 alkyl
`
`esters, and especially isopropyl esters.
`
`Preferred for use in the methods and compositions of the present invention are
`
`those compounds of formula I above, wherein:
`
`R1 = COZR, where R = H or COZR = ophthalmically acceptable ester moiety;
`
`n = 0;
`
`---- = single or non—cumulated double bond, with the provisos that a double bond between
`
`carbons 4 and 5 may not be of the trans configuration; and that a double bond between
`
`carbons l3 and 14 may not be of the cis configuration;
`
`
`
`D = fluorine in the alpha (or) configuration, and D1 = H in the beta (B) configuration;
`
`X = CH2O or CHZCHz; and
`
`Y = phenyl, optionally substituted with halo or trihalomethyl.
`
`Especially preferred are those preferred compounds of formula I above, wherein:
`
`R1 = COZR and COZR = lower alkyl (i.e., 1—6 carbons) carboxylic acid alkyl ester.
`
`Included in these especially preferred compounds are the following novel compounds:
`
`-6-
`
`Micro Labs Exhibit 1012-8
`
`Micro Labs Exhibit 1012-8
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`Compound Number
`
`Compound Name
`
`Compound Structure
`
`(5Z)—(9S,11R,15R)-16-(3-
`Chlorophenoxy)-9, l l-dihydroxy-
`lS-fluoro-17,18,19,20-tetranor—5-
`prostenoic acid isopropyl ester
`
`(4Z)-(9S,11R,15R)-16-(3-
`Chlorophenoxy)-9, 1 l-dihydroxy-
`15—fluoro-17,18,19,20-tetranor—4-
`prostenoic acid isopropyl ester
`
`III
`
`(SZ)-(9S,11R,15R)-9,11-
`Dihydroxy-lS-fluoro-16-[3-
`(trifluoromethyl)phenoxy]—
`17,18,19,20-tetranor-5-prostenoic
`acid isopropyl ester
`
`
`
`(5Z)-(9S,11R,15R)-9,11-
`Dihydroxy- l S-fluoro- l 6-
`phenoxy-17,18,19,20-tetranor-5-
`prostenoic acid isopropyl ester
`
`Included within the scope of the present invention are the individual enantiomers
`
`of the title compounds, as well as their racemic and non-racemic mixtures. The individual
`
`enantiomers can be enantioselectively synthesized from the appropriate enantiomerically
`
`pure or enriched starting material by means such as those described below. Alternatively,
`
`they may be enantioselectively synthesized from racemic/non—racemic or achiral starting
`
`materials (Asymmetric Synthesis by J. D. Morrison and J. W. Scott, Eds., Academic Press
`
`Publishers: New York, 1983—1985 (five volumes) and Principles ofAsymmetric Synthesis
`
`by RE. Gawley and J. Aube, Eds., Elsevier Publishers: Amsterdam, 1996). They may
`
`also be isolated from racemic and non—racemic mixtures by a number of known methods,
`
`6. g. by purification of a sample by chiral HPLC (A Practical Guide to Chiral Separations
`
`by HPLC, G. Subramanian, Ed., VCH Publishers: New York, 1994; Chiral Separations by
`
`HPLC, A.M. Krstulovic, Ed., Ellis Horwood Ltd. Publishers, 1989), or by enantioselective
`
`hydrolysis of a carboxylic acid ester sample by an enzyme (Ohno, M.; Otsuka, M. m
`
`Reactions volume 37, page 1 (1989)). Those skilled in the art will appreciate that racemic
`
`-7-
`
`Micro Labs Exhibit 1012-9
`
`Micro Labs Exhibit 1012-9
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`and non-racemic mixtures may be obtained by several means, including without limitation,
`
`nonenantioselective synthesis, partial resolution or even mixing samples having different
`
`enantiomeric ratios.
`
`In the foregoing illustrations, as well as those provided hereinafier, wavy line
`
`attachments indicate either the alpha (0L) or beta ([3) configuration. The carbon numbering
`
`
`
`is as indicated in the structural depiction of formula I, even when n = 2. A hatched line, as
`
`used e.g. at carbon 9, indicates the 0!. configuration. A solid triangular line, as used e.g. at
`
`carbon 12, indicates the B configuration. Dashed lines on bonds, 6. g. between carbons 13
`
`and 14, indicate a single or double bond. Two solid lines between carbons indicate a
`
`double bond of the specified configuration.
`
`In the following Examples l-4, the following standard abbreviations are used:
`
`g = grams (mg = milligrams); mol = moles (mmol = millimoles); 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 "MS" refers to mass spectrometry.
`
`Micro Labs Exhibit 1012-10
`
`Micro Labs Exhibit 1012-10
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`EXAMPLE 1:
`
`Synthesis of II
`
`
`
`Micro Labs Exhibit 1012-11
`
`Micro Labs Exhibit 1012-11
`
`

`

`WO 98/21181
`
`PCT/US97/2067l
`
`A. |3aR, 4R1 1E,3S ), 5R, 6gfil—5-Benzoyloxy-4—l4—1 3-chlorophenoxy1-3-hydroxy-1-
`
`buten l—hexah dro—2H-c clo enta b furan-2—one 2
`
`To a solution of [3aR, 4R(1E), 5R, 6aS]—5-benzoyloxy-4-[4-(3—chlorophenoxy)-3-
`
`oxo-l-buteny1]—hexahydro—2H—cyclopenta[b]finan-2-one (1; for preparation, see published
`
`European Patent Application EP 639563 A2, which is incorporated herein by this
`
`reference) (1.02 g, 2.32 mmol) in THF (10 mL) at -23 °C (bath temperature) was added
`
`dropwise a solution of (+)—B-chlorodiisopinocampheylborane (available from Aldrich
`
`Chemical Co., Milwaukee, Wisconsin) (1.4 g, 4.4 mmol) in THF (10 mL). The mixture
`
`was then warmed to 0 °C (bath temperature) and was quenched after 90 min by the
`
`addition of methanol (10 mL). Saturated NH4C1 was added (35 mL), the mixture was
`
`extracted with ethyl acetate (3 x 40 mL), dried (MgSO4), filtered, concentrated, and
`
`chromatographed on a 30 cm tall x 41 mm diameter silica gel column eluting with 1:1
`
`ethyl acetatezhexane to afford 2 (502 mg, 49%) as well as a mixture of 2 and its epimeric
`
`alcohol (254 mg, 23%).
`
`B. 3aR 4R 3
`
`5R 6a -5-Benzo 10x -4- 4- 3-chloro henox -3-h drox but 1-
`
`hexah dro—ZH—c clo ent
`
`furan—2- ne 3
`
`A solution of 2 (500 mg, 1.14 mmol) and 10% w/w Pd/C (200 mg) in ethyl acetate
`
`(18 mL) was stirred under 1 atm of Hz for 5.5 h, filtered through Celite, and concentrated
`
`to afford 3 (486 mg, 97%).
`
`
`
`mixture of4 and a by-product (44 mg). 13c NMR (CDC13) 5 176.57 (C), 166.04 (C),
`
`£2.
`
`|3aR, 4R1 3R ), 5R, 6&5 -5-Benzoyloxy—4—|4-(3-chlorophenoxy)-3-fluorobutyl|-
`
`hexahydro-2H—cyclopental b furan-2-one (4 1
`
`To a solution of 3 (480 mg, 1.08 mmol) in CHZCIZ (10 mL) at 0 °C (bath T) was
`
`added (diethylamino)su1fur trifluoride (DAST) available from Aldrich Chemical Co.,
`
`Milwaukee, Wisconsin (450 mg, 2.8 mmol). After 3 h, saturated sodium bicarbonate was
`
`added (20 mL), the layers were separated, extracted with CHZCIZ (2 x 20 mL), dried
`
`(MgSO4), filtered, and chromatograhed on an 18 cm tall x 26 mm diameter silica gel
`
`column eluting with 1:1 ethyl acetatezhexane to afford 4 (117 mg, 33 %) as well as a
`
`-10-
`
`Micro Labs Exhibit 1012-12
`
`Micro Labs Exhibit 1012-12
`
`

`

`WO 98/21181
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`PCT/US97/20671
`
`159.03 (C), 134.93 (C), 133.34 (CH), 130.31 (CH), 129.62 (CH), 128.54 (CH), 121.54
`
`(CH), 115.06 (CH), 113.07 (CH), 91.02 (d, J = 171 Hz, CH), 84.27 (CH), 79.80 (CH),
`
`69.60 (d, J = 24 Hz, CH2), 52.26 (CH), 43.59 (CH), 36.96 (d, J = 76 Hz, CH2), 29.34 ((1, J
`
`= 21 Hz, CH2), 28.57 (CH2).
`
`D. [ 321R, 4R1 3R 1, 5R, 6a,S[-4—|4-13-glhlorophenoxy1-3-fluorobutyl l-5-hydroxy-hexahydro-
`
`2H—c clo enta b furan-Z-one 5
`
`To a solution of 4 (117 mg, 0.26 mmol) in methanol (5 mL) was added KZCO3 (57
`
`mg, 0.41 mmol). After 90, min saturated NH4C1 was added (10 mL), the mixture was
`
`extracted with ethyl acetate (3 x 15 mL), dried (MgSO4), filtered, and chromatographed on
`
`
`
`a 16 cm tall x 26 mm diameter silica gel column eluting with ethyl acetate to afford 5 (60
`
`mg, 67%).
`
`E. |3aR= 4R13R), 5R, 6@ |-4-|4-(3—thorophenoxy)-3-fluorobutyl |-5—(tetrahydrop)aan—2—
`
`yloxy )—hexahydro-ZH—cyclopental b |furan-2—one (6)
`
`To a solution of 5 (59 mg, 0.17 mmol) and 3,4-dihydro-2H-pyran (25 mg, 0.30
`
`mmol) in CHZCl2 (2.5 mL) at 0 °C (bath temperature) was added p-toluenesulfonic acid
`
`monohydrate (8 mg, 0.04 mmol). After 1 h, NEt3 was added (0.1 mL), saturated sodium
`
`bicarbonate was added (5 mL), the layers were separated, extracted with CHZCIZ (3 x 5
`
`mL), dried (MgSO4), filtered, and chromatographed on an 11 cm tall x 26 mm diameter
`
`silica gel column eluting with 1:1 ethyl acetatezhexane to afford 6 (59 mg, 82 %).
`
`F. | 3aR, 4R1 3R 1, 5R, 6a,§l-4-|4—13-Chlorophenoxy)-3-fluorobutyll-S-(tetrahydropyaan-2-
`
`yloxy )—hexahydro-ZH-cyclopental b Ifuran-2-ol (7 )
`
`To a solution of 6 (59 mg, 0.14 mmol) in toluene (2 mL) at —78 OC (bath
`
`temperature) was added dropwise a 1.5 M solution of diisobutylaluminum hydride in
`
`toluene (0.14 mL, 0.21 mmol). After 90 min, the reaction was quenched by the addition of
`
`1:1 methanolzethyl acetate (1 mL), warmed to room temperature, added to a saturated
`
`solution of sodium potassium tartarate (4 mL), and stirred until the emulsion broke. The
`
`layers were separated, extracted with ethyl acetate (3 x 5 mL), dried (MgSO4), filtered,
`
`-11-
`
`Micro Labs Exhibit 1012-13
`
`Micro Labs Exhibit 1012-13
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`

`

`WO 98/21181
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`PCT/US97/20671
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`concentrated, and passed through a pipette plug of silica gel eluting with ethyl acetate to
`
`afford 7 (59 mg, 99%), which was used immediately in the following step.
`
`G. (SQ—(95, 1 1R, 15 R)-16-( 3-g :hlorophenoxyI-l 5-fluoro—9—hydroxy-1 1—(tetrahydrop3gan—
`
`2—yloxy)—17,18, 19,20-tetranor-S-prostenoic acid isopropyl ester (8)
`
`To a suspension of (4-carboxybutyl)triphenylphosphonium bromide (205 mg, 0.46
`
`mmol) in THF (2.5 mL) at 0 °C (bath temperature) was added a 1 M solution of potassium
`
`t-butoxide in THF (0.88 mL, 0.88 mmol). After 15 min, a solution of 7 (59 mg) in THF (1
`
`mL) was added. After 1 h, the reaction was quenched by the addition of saturated NH4CI
`
`(10 mL), extracted with ethyl acetate (3 x 10 mL), dried (MgSO4), filtered, and
`
`concentrated to afford an oil. This oil was dissolved in acetone (4 mL), the solution was
`
`cooled to 0 °C (bath temperature), and DBU was added (106 mg, 0.7 mmol). After 20
`
`min, isopropyl iodide was added, and the reaction was allowed to come to room
`
`temperature overnight. The mixture was added to saturated NH4C1 (5 mL), extracted with
`
`ethyl acetate (3 x 5 mL), dried (MgSO4), filtered, concentrated, and passed through a
`
`pipette plug of silica gel eluting with 1:1 ethyl acetatezhexane to afford 8 (147 mg)
`
`contaminated with triphenylphosphine oxide. The sample was used without further
`
`purification in the next step.
`
`
`
`ethyl acetate:hexane to afford II (31 mg, 47% from lactol 7). 13C NMR (CDC13) 5 173.40
`
`- 9S 11R 15 R -16— 3—Chloro henox -9 ll-dih drox —15-fluoro—17 18 19 20—
`
`tetranor—S-prostenoic acid isopropyl ester (11)
`
`To a solution of impure 8 from above (147 mg) in isopropanol (4 mL) was added
`
`12 M HCl (0.8 mL). After 90, min saturated sodium bicarbonate was added (10 mL), the
`
`mixture was extracted with ethyl acetate (4 x 10 mL), filtered, concentrated, and
`
`chromatographed on a 17 cm tall x 10 mm diameter silica gel column eluting with 1:1
`
`(C), 159.18 (C), 134.90 (C), 130.26 (CH), 129.81 (CH), 129.08 (CH), 121.41 (CH),
`
`115.06 (CH), 113.13 (CH), 91.44 (d, J = 172 Hz, CH), 78.65 (CH), 74.61 (CH), 69.91 (d, J
`
`= 23 Hz, CH2), 67.63 (CH), 52.57 (CH), 51.72 (CH), 42.66 (CH2), 34.01 (CH2), 30.05 ((1,
`
`-12-
`
`Micro Labs Exhibit 1012-14
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`Micro Labs Exhibit 1012-14
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`

`

`WO 98/21 181
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`PCT/US97/20671
`
`J = 21 Hz, CH2), 28.86 (CH2), 28.78 (CH2), 26.73 (d, J = 10 Hz, CH2), 24.90 (CH2), 21.82
`
`(CH3). MS, m/z calcd. for C25H3605FC1Na [(M + Na)+], 493; found, 493.
`
`EXAMPLE 2:
`
`Synthesis of III
`
`
`
`which is reacted with Ph3P+(CH2)3C02H Br' in the presence of KOBut in THF, followed
`
`4- rostenoic acid iso ro
`
`l ester III
`
`Wittig condensation of 7 with Ph3P+CHZOMe C1' in the the presence of KOBu' in
`
`THF yields enol ether 9. Acidic hydrolysis using TsOH in THF/water gives lactol 10,
`
`-fluoro-l7 18 19 20-tetranor-
`
`by treatment of an acetone solution of the resulting carboxylic acid with DBU and
`
`isopropyl iodide, to afford III.
`
`Micro Labs Exhibit 1012-15
`
`Micro Labs Exhibit 1012-15
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`Synthesis of IV
`
`
`
`Micro Labs Exhibit 1012-16
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`Micro Labs Exhibit 1012-16
`
`

`

`WO 98/21181
`
`PCT/US97/2067l
`
`A. 3 aR, 4R1 1E,3 S ), 5R: 6& |-5-Benzoyloxy—4-| 3-hydroxy-4-( 3-
`
`(trifluoromethyl )phenoxy)-1-buteny1 -hexahydro—2H—cyclopental b furan-2-one (12)
`
`To a solution of [3aR, 4R(1E), 5R, 6aS]—5-benzoyloxy-4-[3-oxo-4-(3—
`
`(trifluoromethyl)phenoxy)—1-buteny1]-hexahydro-2H—cyclopenta[b]furan-2-one (1 1; for
`
`preparation, see US. Patent Number 4,321,275, which is incorporated by this reference)
`
`(1.77 g, 3.8 mmol) in THF (22 mL) at 0 °C (bath temperature) was added a solution of
`
`—(+)-B-chlorodiisopinocampheylborane (2.59 g, 8.1 mmol) in THF (37 mL). After 1 h, the
`
`reaction was warmed to room temperature, and, after 1 additional h, saturated sodium
`
`bicarbonate (30 mL) was added. The solution was extracted with ethyl acetate (3 x 30
`
`mL), dried (MgSO4), and filtered, and the residue was dissolved in 150 mL of 1:2
`
`acetonitn'lezhexane. The two-phase mixture was shaken in a separatory funnel and the
`
`bottom layer was concentrated. The residue was chromatographed on a 28 cm tall x 41
`
`mm diameter silica gel column eluting with 3:2 ethyl acetate:hexane to afford 12 (624 mg,
`
`35%) as well as a mixture of 12 and the corresponding epimeric alcohol (681 mg, 39%).
`
`B. |3aR, 4R13S), 5R, 6&3 l-5—Benzoyloxy-4—l3-hydroxy-4-13—(trifluoromethyl)phenoxy)-
`
`butyl —hexahydro-2H-cyclopental b Ifuran-Z—one ( 13)
`
`A solution of 12 (600 mg, 1.29 mmol) and 10% Pd/C (77 mg) in ethyl acetate (15
`
`mL) was stirred under 1 atm of H2 for 3.5 h, filtered through Celite, and concentrated to
`
`afford 13 (601 mg, 100% yield).
`
`
`
`with 4:1 diethyl etherzhexane to afford 14 (312 mg, 50 %). 13C NMR (CDC13) (partial
`
`C. I3aR, 4R13R), 5R, 6a,§|-5-Benzoyloxy-4- 3—fluoro-4-(3-
`
`(trifluoromethyl )phenoxy )butyl |—hexahydro-2H—cyclopenta| b Ifuran-Z-one 1 14)
`
`To a solution of 13 (600 mg, 1.29 mmol) in CHZCIZ (10 mL) at 0 °C (bath T) was
`
`added DAST (340 mg, 2.1 mmol). After 50 min, saturated sodium bicarbonate was added
`
`(15 mL), the layers were separated, extracted with CHZCIZ (2 x 15 mL), dried (MgSO4),
`
`filtered, and chromatograhed on an 30 cm tall x 41 mm diameter silica gel column eluting
`
`spectrum) 6 91.10 (d, J = 172 Hz, CH), 84.32 (CH), 79.88 (CH), 69.67 (d, J = 23 Hz,
`
`-15-
`
`Micro Labs Exhibit 1012-17
`
`Micro Labs Exhibit 1012-17
`
`

`

`WO 98/21181
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`PCT/US97/20671
`
`CH2), 52.25 (CH), 43.55 (CH), 36.93 (d, J = 75 Hz, CH2), 29.36 (CH2), 28.71 (d, J = 11
`
`Hz, CH2), 28.41 (CH2).
`
`D. 3aR, 4R13R), 5R, 6a,: |-4—| 3-Fluoro-4-( 3-1 trifluoromethyl )phenoxy )butyl [-5-hydroxy-
`
`5
`
`hexahydro—ZH—cyclopental b lfuran—2—one ( 15)
`
`To a solution of 14 (310 mg, 0.64 mmol) in methanol (10 mL) was added KZCO3
`
`(128 mg, 0.93 mmol). After 2.5 h, saturated citric acid was added (20 mL), the mixture
`
`was extracted with ethyl acetate (3 x 20 mL), dried (MgSO4), filtered, and
`
`chromatographed on a 14 cm tall x 26 mm diameter silica gel column eluting with 3:2
`
`ethyl acetatezhexane to afford 15 (197 mg, 82%).
`
`E. |3aR, 4R13R), 5R, 6a§ -4— 3—Fluoro-4-l3-(trifluoromethyl)phenoxy)butyl|-5-
`
`
`
`tetrah dro
`
`an—2-
`
`lox —hexah dro—2H—c clo enta b furan—2—one 16
`
`To a solution of 15 (191 mg, 0.51 mmol) and 3,4—dihydro—2H—pyran (64 mg, 0.77
`
`mmol) in CH2C12 (3.2 mL) at 0 °C (bath temperature) was added p-toluenesulfonic acid
`
`monohydrate (60 mg, 0.31 mmol). After 25 min, NEt3 was added (0.2 mL), saturated
`
`sodium bicarbonate was added (20 mL), the layers were separated, extracted with ethyl
`
`acetate (2 x 20 mL), dried (MgSOA), filtered, and chromatographed on an 14 cm tall x 26
`
`mm diameter silica gel column eluting with 3:2 ethyl acetatezhexane to afford 16 (224 mg,
`
`95 %).
`
`F. |3aR, 4R13R), 5R, 6a§|—4-| 3-Fluoro—4—1 3-( trifluoromethyl )phenoxy )butyl |—5-
`
`tetrah dro
`
`ran-2— lox
`
`-hexah dro-2H—c clo enta b furan-2-ol 17
`
`To a solution of 16 (220 mg, 0.48 mmol) in toluene (4 mL) at - 78 °C (bath
`
`temperature) was added dropwise a 1.5 M solution of diisobutylaluminum hydride in
`
`toluene (0.48 mL, 0.72 mmol). After 30 min, the reaction was quenched by the addition of
`
`1:1 methan012ethyl acetate (3 mL), warmed to room temperature, added to a saturated
`
`solution of sodium potassium tartarate (20 mL), and stirred until the emulsion broke. The
`
`layers were separated, extracted with ethyl acetate (2 x 20 mL), dried (MgSO4), filtered,
`
`-16-
`
`Micro Labs Exhibit 1012-18
`
`Micro Labs Exhibit 1012-18
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`

`

`WO 98/21181
`
`PCT/US97/20671
`
`and concentrated to afford 17 (220 mg, 100%), which was used immediately in the
`
`following step.
`
`— 9S 11R 15 R ~15—F1uoro-9-h drox -11- tetrah dro
`
`trifluorometh 1 henox -17 18 19 20-tetranor-5- rostenoic acid iso r0
`
`1 ester 1
`
`To a suspension of (4-carboxybutyl)tripheny1phosphonium bromide (750 mg, 1.7
`
`mmol) in THF (5 mL) at 0 °C (bath temperature) was added a 1 M solution of potassium
`
`t-butoxide in THF (3.0 mL, 3.0 mmol). After 10 min, a solution of 17 (220 mg) in THF (3
`
`mL) was added. After 90 min, the reaction was quenched by the addition of saturated
`
`7 NH4C1 (20 mL), extracted with ethyl acetate (3 x 20 mL), dried (MgSO4), filtered, and
`
`concentrated to afford an oil. This oil was dissolved in acetone (11 mL), the solution was
`
`cooled to 0 °C (bath temperature), and DBU was added (380 mg, 2.5 mmol). After 10
`
`min, isopropyl iodide was added, and the reaction was allowed to come to room
`
`temperature overnight. The mixture was added to saturated NH4C1 (15 mL), extracted
`
`with ethyl acetate (3 x 20 mL), dried (MgSO4), filtered, concentrated, and
`
`chromatographed on a 14 cm tall x 26 mm diameter silica gel column eluting with 3:2
`
`hexanezethyl acetate to afford 18 (147 mg, 52%).
`
`
`
`ethyl acetate gradient to afford IV (92 mg, 73%). 13C NMR (CDC13) (partial spectrum) 5
`
`H. (52H9S, 11R, 15 R)-9,11—Dihydroxy-15—fluoro-16-l3-1trifluoromethyljphenoxyl~
`
`1 7,1 8,19,20—tetranor—5—prostenoic acid isopropyl ester (IV)
`
`To a solution of 18 (146 mg) in isopropanol (6 mL) was added 12 M HCl (0.4 mL).
`
`After 3.5 h, saturated sodium bicarbonate was added (10 mL), the mixture was extracted
`
`with ethyl acetate (2 x 10 mL), filtered, concentrated, and chromatographed on a 17 cm tall
`
`x 26 mm diameter silica gel column eluting with a 3:2 ethyl acetatezhexane to straight
`
`91.44 ((1, J = 172 Hz, CH), 78.60 (CH), 74.54 (CH), 69.97 ((1, J = 23 Hz, CH2), 67.64
`
`(CH), 52.49 (CH), 51.70 (CH), 42.66 (CH2), 34.01 (CH2), 30.01 (d, J = 21 Hz, CH2),
`
`28.84 (CH2), 28.77 (CH2), 2670 (d, J = 9 Hz, CH2), 24.89 (CH2), 21.79 (CH3). MS, m/z
`
`calcd. for C26H3705F4 [(M + H)+], 505.25706; found, 505.25705.
`
`-17-
`
`Micro Labs Exhibit 1012-19
`
`Micro Labs Exhibit 1012-19
`
`

`

`W0 98/21 181
`
`PCT/US97/20671
`
`EXAMPLE 4:
`
`
`
`Synthesis of V
`
`Micro Labs Exhibit 1012-20
`
`Micro Labs Exhibit 1012-20
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`W A
`
`solution of [3aR,4R(1E),5R,6aS]-5-benzoyloxy-4-formyl-hexahydro-ZH—
`
`cyclopenta[b]furan-2—one (19) (available from Cayman Chemical Company, Ann Arbor,
`
`Michigan) (8.00 g, 29.2 mmol) in dichloromethane (50 mL) was added to a 0 °C (bath
`
`temperature) suspension of dimethyl (3-phenoxy-2-oxopropyl)phosphonate (prepared in a
`
`manner analogous to that described in US. Patent No. 5,665,773 for dimethyl (2-oxo—3-(3-
`
`chlorophenoxy)propyl)phosphonate, which patent is incorporated herein by this reference)
`
`(8.29 g, 32.1 mmol), lithium chloride (1.37 g, 35.04 mmol) and triethylamine (4.64 ml,
`
`33.3 mmol) in THF (100 mL) with mechanical stirring for three hours. The reaction was
`
`quenched with aqueous 2 N hydrochloric acid (50 mL) and water (100 mL). The mixture
`
`was extracted with dichloromethane (200 ml and then 100 mL). The combined organic
`
`layers were washed with brine (100 mL), dried over magnesium sulfate and evaporated to
`
`a yellow oil. The resulting residue was purified by chromatography using silica gel and
`
`dichloromethane/ethyl acetate (8:2) to afford 20 as a white solid, 9.17 g (mp 120—5 °C ).
`
`MS, m/z calcd. for C24H2206Na [(M+Na)+], 429; found, 429
`
`2H—cyclopental b Ifuran-Z—one (21 1
`
`A solution of 20 (8.16 g, 20 mmol) in THF (100 mL) was cooled to 0 °C (bath
`
`temperature) and treated with (+)-B—chlorodiisopinocampheylborane (12.8 g, 40 mmol).
`
`The reaction was warmed to room temperature and stirred overnight. The reaction was re-
`
`cooled to 0 °C and quenched with acetone (44 ml, 600 mmol) with stirring for one hour.
`
`Then methanol (50 mL) was added and the mixture was stirred for fifteen minutes. After
`
`the solvent was evaporated the residue was taken up in acetonitrile (100 mL) and washed
`
`with hexane (100 mL). The acetonitrile solution was passed through a pad of silica gel
`
`and evaporated, and the residue was purified by chromatography on silica gel eluting with
`
`hexane/ethyl acetate/dichloromethane (35:65:5) to afford 21 as a colorless oil (1.56 g,
`
`19%). MS, m/z calcd. for C24H24O6 [(M+Na)+], 431; found, 431.
`
`
`
`B. I 3aR,4R1 71E,3§ ),5R,6a§ |~5—( Benzoyloxy)—4—| 3-hydroxy—4-phenoxybuteny1 -hexahydro-
`
`-19-
`
`Micro Labs Exhibit 1012-21
`
`Micro Labs Exhibit 1012-21
`
`

`

`WO 98/21181
`
`PCT/US97/20671
`
`cyclopenta b |furan~2~one (22)
`
`A solution of 21 (1.55 g, 3.8 mmol) in ethyl acetate (50 mL) was treated with 10%
`
`Pd/C (200 mg) with shaking under hydrogen (50~60 psi) for 8 hours. The reaction was
`
`-hexah dro-ZH-
`
`g, 97%). MS, m/z calcd. for C24H2406Na [(M+Na)+], 433; found, 433.
`
`filtered through celite and the solvent was evaporated to provide 22 as a colorless oil (1.51
`
`
`
`D. | 3aR,4R( 3R ), 5R,6a§ |-5-1 Benzoyloxy )—4-| 3-fluoro-4-phenoxybutyl I-hexahydro-ZH-
`
`cyclopental b lfuran-Z-one (23)
`
`A 0 °C (bath temperature) solution of 22 (1.50 g, 3.65 mmol) in dichloromethane
`
`(40 mL) was treated with DAST (0.72 ml, 5.48 mmol) an