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`U.S. PATENT APPLICATION
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`60/030, 519
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`PAUL H. ZINE, FORT WORTH, TX.
`
`g Pmn G. KLIHKO, FORT WORTH, TX: MARK R. NERO,
`
`0 5 FEB 1998
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`eemxmnqc “unannnnn"an...
`VERIFIED
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`“FOREIGN/PCT APPLICATIONS""" "u"
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`BARRY L COPELAND
`FARM DEPT Q 148 MOON LABORATORIES INC
`5201 SOUTH menu
`NR! WORTH TX 16134-2099
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`ls-nuono PROSTAGMNDINS AS OCULAR mmsms
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`IS-FLUORO PROSTAGLANDINS AS OCULAR HYPOTENSIVES
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`lS-FLUORO PROSTAGLANDINS AS OCULAR HYPOTENSIVE
`
`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
`
`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 intraoculsr 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 (“10?”) 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 tennination of therapy.
`
`Systemically administered carbonic anhydrase inhibitors can also cause serious side efl'ects,
`
`such as nausea, dyspepsia, fatigue, and metabolic acidosis, which side effects can affect
`
`patient compliance and/or necessitate the termination of treauuent. Some beta-blockers
`
`have increasingly become associated with serious pulmonary side effects attributable to
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`Micro Labs Exhibit 1013-4
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`their effects on beta-2 receptors in pulmonary tissue. Sympathomimetics may cause
`
`‘ tachycardia1 arrhythmia and hypertension. There is therefore a continuing need for
`- therapies which control the elevated intraocular pressure associated with glaucoma.
`
`
`
`s
`
`‘
`
`Prostaglandins, which are metabolite derivatives of arachidonic acid. have recently
`
`been pursued for possible efficacy in lowering IOP. Arachidonic acid in the body is
`
`convened to prostaglandin 62, which is subsequently convened to prostaglandin H2. Other
`
`naturally occurring prostaglandins are derivatives of prostaglandin Hz. A number of
`
`5'
`
`different types of prostaglandins have been discovered including A, B, D, E, F. G. i and J—
`
`to
`
`Series prostaglandins (EPO 561 073 Al). or interest in the present invention are
`
`compounds which are believed to exhibit IOP lowering effects similar to those exhibin by
`
`PGqu (an F—series prostaglandin):
`
`The relationship of P65. receptor activation and IOP lowering effects is not well
`
`
`
`understood. It is believed that PGcm receptor activation leads to increased outflow of
`aqueous humor. Regardiess omeZe-chanism, PGF;a and analogs have‘been shown to lower
`ror> (Giuffre, 11.: 5mm afProrrngbndin a. the Human Eye,mm;
`
`Qahthalmology, volume 222. pages l39-l4l (1985); and Kerstetter et al.. Prosraglandin
`
`
`
`
`
`
`
`
`20
`
`
`
`
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`
`
`rs
`
`been of interest in the field to develop synthetic PGqu analogs with IOP lowering efficacy.
`
`Synthetic PGFh-typc analogs have been pursued in the artW
`
`9911mm. volume 229, pages 4| 1-413 (1991)). Though PGFm-type molecules lower
`
`Micro Labs Exhibit 1013-5
`
`‘
`
`F1.1.] -Iropropylester lowers Intraocular Pressure Without Decreasing Aqueous Humor
`
`FIDW. mmmmnamm. volume “)5. Pages 3034 (1988)). Tim. it has
`
`Micro Labs Exhibit 1013-5
`
`

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`
`
`The present invention is directed to compositions and methods of their use in
`
`treating 101’ and ocular hypertension. In particular. the present invention provides l5-
`
`fluoro prostaglandin analogs believed to have functional PGFz. receptor agonist activity,
`
`and methods of their use in treating glaucoma and ocular hypertension. As previously
`stated, the mechanism of action by which PGFz. type prostaglandins lower 101' is not well
`
`'
`
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`
`1
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`so
`
`understood While the mechanism of action of the compounds of the present invention is
`
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`Micro Labs Exhibit 1013-6
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`10?, many of these types of molecules have also been associated with urnlesirable side
`
`_ effects resulting from topical ophthalmic dosing. Such effects include an
`
`increase in
`
`‘ IOP, breakdown of the blood aqueous barrier and conjunctival hyperemia (Alm. 771:
`. Eoteritia! ofPrastaglandin Derivatives in Glaucoma Therapy.
`5 mm, volume 4. No. 11. pages 44-50 (1993)).
`
`Based on the foregoing, it need exists for the development of molecules that may
`
`activate f’GFz.I receptors, yielding a more efficacious lowering of 10?. while exhibiting
`
`fewer or reduced side effects.
`
`to
`
`‘ ' '
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`L '
`
`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 object of this invention to provide a class of [OP lowering agents with an improved
`
`therapeutic profile over PCP“. and methods of their use. It has now unexpectedly been
`
`is
`
`discovered that the presently claimed 15-fluoro analogs of PGFI. meet this objective.
`
`While some prostaglandins with fluorine in the omega chain are known in the an (B?
`
`435,443 A; 1? 7,070,054 A2;MW, volume 57. number 2. pages 39-41
`
`(1994) (Cheuu'mlAhsmts. volume 121. abstract 50656 (1994));WEE.
`
`m. volume 6. pages 831-7 (1989)mm. volume 112, abstract 30749
`
`20
`
`(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.
`
`V1,.Wu".1
`
`Micro Labs Exhibit 1013-6
`
`

`

`
`
`not fully understood. the inventors theorize that such compounds exhibit enhanced FP
`
`I
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`r
`
`_ receptor selectivity as a consequence of their decreased activity at the E? 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.
`
`2.!m..“1|.
`
`found that lS-fluom substituted PGFuanalogs of the
`It has unexpecmdly
`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 PGFz. analogs of the present invention have the following
`formula I:
`
`320’,
`"
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`R35
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`wherein:
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`R' =C02R, CONR‘R’. CH10R30I’CH1NR7R'. where
`R = H or cationic salt moiety,_gr—C01R = pharmaceutially acceptable ester moiety; R'.
`RS = same or different = H or alkyl; R“ = H, acyl. or alkyl; R’, R‘ = same or different =
`
`H, acyl. or alkyl; with the proviso that if one of 11'. R' = acyl. then the other = H or
`
`alkyl;
`
`n = 0 or 2;
`
`—— = single or non-cumulated double bond, with the provisos that a double bond between _
`
`V
`
`carbons 4and 5 may notheoftheuamconfigmafiomand thatadouble bond between
`
`carbons l3 and 14 may not be ofthe ci: configuration;
`
`‘ IMicro Labs Exhibit 1013-7
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`Micro Labs Exhibit 1013-7
`
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`x = (cagq or (cagqo; where q = 1-6; and
`
`Y = a phenyl ring optionally substituwd with alkyl. halo, uihalomcthyl, alkoxy, acyl,
`acyloxy, amino, alkylamino. acylamino, or hydmxy; or
`
`X-Y = (CH2)PY'; whererp =' 0-6; and
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`w =Cl-lz, 0, 8(0).. NR’, CH1Cl-l2, CH=CH, CH,o. 0123(0)... CH=N. or
`CHzNR’; wherem =o-2, and R’=H, alkyl. oracyl;
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`Z= H. alkyl,alkoxy, acyl,acyloxy, halo,u'ihalomethyl,amino,alkylamino.
`acylamin'ov or'h di'oxy; and
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`-— = single or double bond.
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`For purposes of the {ongoing definition, the term “phannaceuu'cally acceptable
`eaten" means any ester that would be suitable for therapeutic administration to a patient by
`any conventional means without significant deleterious health conseqmncm; and
`“ophthalmically aooeplable ester” means any pharmaceutically acceptable can that would
`
`be suitable for ophthalmic application, i.c. non—toxic and non-inhaling. Ptefeaed among
`
`
`
`Micro Labs Exhibit 1013-8
`
`

`

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`
`the ophthalmically acceptable essters are alkyl esters. Most preferred are C1-C4 alkyl
`
`esters. and especially isopropyl esters.
`
`.
`
`. Preferred for use in the methods and compositions of the present invention are
`compounds of formula 1 above. wherein:
`
`R‘ = C0112, where R = H or C02R = ophthalmically acceptable ester moiety;
`
`n=0;
`
`5
`
`IO
`
`~~ = single or non-emulated 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;
`
`is R2=R3=H;
`
`D = fluorine in the alpha (:1) configuration. and D' = H in the beta (a) configuration;
`
`X = CH20 or CHzCHz: and
`
`Y = phenyl. optionally sulgstituted with halo or nihalomethyl.
`
`Especially preferred are those preferred compounds of {canola 1 above. wherein: R'
`
`= COIR and com = lower alkyl (i.e.. 1-6 carbons) carboxylic acid alkyl esmr. lnriuded in
`
`these especially preferred compounds are the following novel compounds:
`
`20
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`Micro Labs Exhibit 1013-9
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`Micro Labs Exhibit 1013-9
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`
`
`Compound Name
`(52)-(9S,llR.lSR)—l&(3-
`Chlmophemny)9.l l-dihydroxy-
`lS-flml1,18,19.20-tetranor-5-
`prostenoic acid isopmpyl cam
`
`(4Z)—(9S.11R.l5R)-16-(3-
`Chlorophenoxyw, l l-dihydroxy-
`lS-fluoro-l7,18.l9.20-tetranor-4-
`prostcnoic acid isopropyt ester
`
`acid isopropyl ester
`
`(SD-(95.1 1R,15R)-9,l lo
`Dihydmxy-lS-fluoro-lG-B-
`(trifluoromethyl)pbenoxy]—
`l7, 18.19.20-tetmnor-5-postenoic
`
`Compound Number
`
`ompound Structure
`
`“Qt ,WP',
`
`In the foregoing illustrations. as well as those provided hereinafter. wavy line
`
`attachments indicate either the alpha (0.) or beta (B) 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 a configuration. A solid triangular line, as used eg. at
`
`carbon 12. indicates the B configuration. Dashed lines on bonds, eg. between carbons l3
`
`and 14, indicate a single or double bond. Two solid lines between carbons indicate a
`
`double bond of the specified configuration.
`
`In the following-1:2. the following standard abbreviations are used:
`g = grams (mg = milligrams): mol = molm (mmol = millimoles); mL = milliliters;
`
`mm Hg = millimeters of mercury; mp = melting point; bp = boiling point; it = hours; and
`
`min = minutes. In addition. 'NMR" refers to nuclear magnetic resonance spectroscopy and
`
`'MS' refers to mass spectrometry.
`
`Micro Lab§ Exhibit 1013-10
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`Micro Labs Exhibit 1013-10
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`Synthesis of II
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`Mi'cko Lab; Exhibit 1013-11
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`Micro Labs Exhibit 1013-11
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`- To a solution of [3aR. 4R(1E). 5R. 6aS].5-benzoyloxy-4-[4—(3-chlorophenoxy)~3-
`'oxo‘lLbutenyl]—hexahydro—2H-cyclopenta[b]furan-Z-onc (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
`
`IO
`
`dropwise a solution of (+)-B-chlomdiisopinocampheylborane (1.4 g, 4.4 mmol) in THF (10
`mL). The mixture
`warmed to 0 °C (bath temperature) and was quenched after 90
`min by the addition Biimemanol (10 mL). Saturated NH4CI was added (35 mL). the
`
`mixture was extracted with ethyl acetate (3 x 40 mL). dried (Mgso.). 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%).
`
`15
`
`B
`
`4
`
`R
`
`- .
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`
`-4- 4-
`
`_
`
`A solution of 2 (500 mg. 1.14 mmol) and 10% w/w MC (200 mg) in ethyl acetate
`
`(l8 ml.) was stirred under 1 atm of H; for 5.5 h. filtered through Celite. and concentrated
`
`to afford 3 (486 mg, 97%).
`
`
`exalt
`n i
`i
`n
`4
`
`To a solution of 3 (480 mg. 1.08 mmol) in CH1Cl1 (10 ml.) at 0 °C (bath 1') was
`
`added (diethylamino)sulfur trifluoride (DAS'D (450 mg, 2.8 mmol). After 3 h, saturated
`sodium bicarbonate was added (20 mL).vthe layers were separated. extracted with GM];
`(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 (1 17 mg. 33 %)
`
`as well as a mixture of 4 and a lay-product (44 mg). "C NMR (CD03) 8 176.57 (C).
`
`166.04 (C). 159.03 (C). 134.93 (C), 133.34 (CH). 130.31 (CH), 129.62 (CH). [28.54
`
`(CH). 12L54 (CH). 115.06 (CH). 113.07 (CH), 91.0201, 1 = 171 Hz, CH). 84.27 (CH),
`
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`Micro Labs Exhibit 1013-12
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`Micro Labs Exhibit 1013-12
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`

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`,ga,g_.<m“y..-.t.“na'i-'fl‘rr'v-uaznI“
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`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 (d. J = 21 Hz. CH2). 28.57 (CH2).
`
`4
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`
`To a solution of 4 (l 17 mg, 0.26 mmol) in methanol (5 ml.) was added K1C03 (57
`
`mg, 0.41 mmol). After 90. min saturated NH.CI was added (10 mL). the mixture was
`
`extracted with ethyl acetate (3 x 15 mL), dried (MgSOd. filtered, and chromatographed on
`a 16 cm tall x 26
`diameter silica gel column eluting with ethyl acetate to afford 5 (60
`IO mg. 67%).
`
`[5
`
`"m.
`
`
`
`23:; w4
`X
`‘
`n
`‘ ‘
`. To a solution of 5 (59 mg, 0.17 mmol) and 3.4—dihydro-2H—pyran (25 mg. 0.30
`mmol) in CHzClz (2.5 ml.) at 0 °C (bath temperature) was addedp-toluenesulfonic acid
`monohydrate (8 mg, 0.04 mmol). After 1 h, NEt, was added (0.1 mL), saturated sodium
`bicarbonate was added (5 mL). the layers were separated, extracted with (:qu1 (3 x 5
`mL). dried (MgSO.), filtered, and chromatographed on an I] cm tall x 26 mm diameter
`silica gel column eluting with H ethyl acetatezhexane to afford 6 (59 mg. 82 96).
`_ >
`I
`p g
`:_
`.
`.
`.
`,
`F, mg, 4R(3R), 5k,agents-gmgmhgggxfl-a-nggmm:1-5-gggmydmxm-g-
`-h x
`i
`n
`-
`-
`l
`To a solution of 6 (59 mg. 0.14 mmol) in toluene (2 ml.) at ~78 °C (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
`l:l methanolzethyi 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
`
`5 1.iii“S
`
`.
`'i
`'Ie".4‘
`I"?
`.-.'
`(2'7‘aa’5 n‘
`9.1.4
`
`.
`
`1'23;
`a
`{25:35
`
`4;
`
`10
`
`25
`
`layers were separated, extracted with ethyl acetate (3 x 5 mL). dried (M3504). filtered, r
`
`concentrated, and passed through a pipette plug of silica gel eluting with ethyl acetate to
`
`30
`
`afford 7 (59 mg, 99%), which was used immediately in the following step.
`
`-10-
`
`
`
`461.353.
`
`n
`
`Micro Labs, Exhibit 1013-13
`
`Micro Labs Exhibit 1013-13
`
`

`

`
`
`
` . To a suspension of (4-carboxybutyl)triphenylphosphonium bromide (205 mg. 0.46
`
`inmol) in THF (2.5 ml.) at 0 °C (bath temperature) wu added a 1 M solution of potassium
`
`r-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 NH.C|
`
`(10 mL). extracted with ethyl acetate (3 x 10 mL). dried (MgSOJ. 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 ovemight. The mixture was added to saturated NH4C1(5 mL), extracted with
`
`ethyl acetate (3 x 5 mL). dried (MgSOr). filtered. concentrated. and passed through a
`
`pipette plug of silica gel eluting with 1:1 ethyl acetatezhexane to afford 8 (147 mg)
`
`contaminated with u-iphenylphosphine oxide. The sample was used without further
`
`purification in the next step.
`
`.2_
`
`.
`
`.l.
`
`I
`
`E
`
`To a solution of impure 8 from above (147 mg) in isopropanol (4 mL) was added
`
`.1W/Jd‘W.--
`
`..1_slrlf.I:"It!
`
`‘2.
`
`5 a
`
`
`
`'.at..mullfllit
`
`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
`
`chromamgraplied on a 17 cm tall x 10 mm diameter silica gel column eluting with 1:1 ethyl
`
`acetatezhexane to afford-'11 (31 mg. 47% from lactol 7). "C NMR(CDC1;) 8 173.40 (C),
`159.13 (C). 134.90(C).130.2(57(CH):129.81 (CH), 129.03 (CH). 121.41 (CH). 115.06
`(CH). 113.13 (CH). 91.44 (d. l = 172 Hz. CH). 78.65 (CH), 74.61 (Cl-1), 69.91 (d. J = 13
`
`Hz. CH2). 67.63 (CH). 52.57 (CH). 51.72 (CH). 42.66 (CH2). 34.01 (CH2). 30.05 (d. .l =
`
`21 Hz. CH2). 28.86 (CH2). 23.78 (CH2). 26.73 (d, .1 = 10 Hz. CH2). 24.90 (CH2). 21.82
`
`(CH;). MS. mlz calcd. for ConDgFClNa [(M + Na)‘], 493; found. 493.
`
`
`
`..._--~va~‘‘
`
`
`
`—»
`
`Micro Labs Exhibit 1013-14
`
`Micro Labs Exhibit 1013-14
`
`

`

`
`
`-
`
`EXAMPLE 2:
`
`'
`
`Synthesis of m
`
`32
`go.
`HQ“- 3‘9“.
`We .—_.
`
`E
`7
`
`Cl
`
`THP6
`
`F
`9
`
`—.
`
`Cl
`
`a
`
`mp5
`
`HO
`
`"a
`
`=
`H6
`
`5
`
`"Qt.
`
`,o‘V=\/\c D P ,
`
`-
`T:
`10
`
`° —’ 5
`H5
`
`C'
`
`5
`i
`ll
`
`4 -
`
`-
`
`-
`
`-
`
`1
`
`-
`
`
`
`Witn'g condensation of 7 with PthGlzoMe Cl‘ in the the pmence of KOBu' in
`THF yields enol ether 9. Acidic hydrolysis using TsOH in THE/water gives lactol 10,
`
`ID
`
`which is reacted with Pth‘(CH2)3COzl-l Bi" in the presence of KOBu‘ THF, followed by
`treatment of an acetone-solution 'ohhe resulting carboxylic acid with .DBU and isopropyl
`iodide, to afford m.
`
`.12.
`
`"3'
`
`_. .--
`
`1'
`
`-
`
`'
`
`A
`
`-
`
`-'-".~- =Z'-
`
`sill-1:? Rift-"'5
`
`‘3
`
`'.' =z'.-i".‘=.‘:' ~23.-
`
`‘
`
`-
`
`-
`
`'v .
`
`-
`
`Micro Labs Exh‘ibit .1013L15
`
`Micro Labs Exhibit 1013-15
`
`

`

`
`
`Micro Labs Exhibit 1013-16
`
`

`

`
`
`
`
`,.nr-l-"
`‘o‘.I—-',~( A
`
`fl."
`....-:‘~
`
`
`
`rr1«5%'IJ'JJ
`
`gag
`
`
`
` _
`
`-' To a solution of Bali. 4R(1E). 5R, oasl-S—benzoyloxy-4-[3-oxo-4-(3-
`(trifluoromethyl)phenoxy)—1~butenyl]~hexahydro-2H-cyclopentaIblfirran-2-one (11; 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 (MgSOa), and filtered. and the residue was dissolved in 150 ml. of 1:2
`
`acetonitrilezhexane. The two-phase mixture was shaken in a sepamtory 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%).
`
`
`
`A solution of 12 (600 mg, 1.29 mmol) and 10% PdIC (77 mg) in ethyl acetate (15
`
`ml.) was stirred under 1 atm of Hz for 3.5 h, filtered through Celite, and concentrated to
`
`afford 13 (601 mg, 100% yield).
`
`
`
`To a solution of 13 (600 mg, 1.29 mmol) in CH1Clz (10 m1.) at 0 °C (bath ’1') was
`
`added DAST (340 mg. 2.1 mmol). After 50 min, saturated sodium bicarbonate was added
`
`(15 mL). the layers were separated. extracted with CH2C1; (2 x 15 mL), dried (Mgsoo,
`
`filtered. and chromatograhed on an 30 cm tall x 41 mm diameter silica gel column eluting
`
`with 4:1 diethyl ethenhexane to afford 14 (312 mg, 50 96). "C m (coca) (partial
`
`specmun) 8 91.10 (d, J = 172 Hz. CH). 84.32 (CH). 79.88 (CH). 69.67 ((1, J = 23 Hr.
`
`CH2). 52.25 (CH). 43.55 (CH). 36.93 (d. J = 75 Hz. CH2). 29.36 (CH2), 28.71 (d, J = 11
`
`Hz. CH1). 28.41 (CH;).
`
`44’
`
`.o ati‘l'u ‘H
`‘0 V‘.‘
`mu'
`.3...‘ .3“: 3:4 .41.. 529.19.. ". I.‘a.1¢1‘y'h\..t M‘hl‘ NU -
`
`his.“ new "Raw.
`‘
`
`—-
`
`Micro LabsExhibit 1013-17
`
`
`
`
`
`Micro Labs Exhibit 1013-17
`
`

`

`
`
` i
`
`grist.
`
`
`
`
`
`- To a solution of 14 (310 mg, 0.64 mmol) in methanol (10 ml.) was added K2CO:
`
`(128 mg, 0.93 mmol). After 2.5 h. saturated citric acid was added (20 mL), the mixture
`
`5
`
`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%).
`
`IO
`
`To a solution of 15 (191 mg, 0.51 mmol) and 3,4-dihydro-2H~pyran (64 mg, 0.77
`
`mmol) in Cl-lzflz (3.2 mL) at 0 °C (bath temperature) was added p-toluenesulfonic acid
`
`monohydrate (60 mg, 0.31 mmol). After 25 min, NEt; was added (0.2 mL). saturated
`
`sodium bicarbonate was added (20 mL), the layers were separated. extracted with ethyl
`
`ts
`
`acetate (2 x 20 mL), dried (MgSOd, 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 %).
`
`
`
`To a solution of 16 (220 mg. 0.48 mmol) in toluene (4 ml.) at — 78 °C (bath
`
`temperature) was added'dropwiséa 1.5 M solution of diisobutylaltuninum hydride in
`toluene (0.48 mL, 0.72
`inertia min, the reaction was quenched by the addition of
`1:1 methanolzethyl acetate (3 mL), warmed to room temperature, added to a saturated
`
`2.5
`
`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 (Mgsoo. filtered.
`
`and concentrated to afford 17 (220 mg. 100%). which was used immediately in the
`
`(
`
`.IQ‘
`'5‘
`
`3
`
`We?”I
`‘a'u
`‘NI"l
`'0
`ha
`fl!' ..3.
`,o.
`
`or.
`
`
`
`following step.
`
`Micro Labs Exhibit 1013-18
`
`‘
`
`Micro Labs Exhibit 1013-18
`
`

`

`
`
`
`
`mL) was added. After 90 min the reaction was quenched by the addition of saturated
`
`NH4C1 (20 mL), extracted with ethyl acetate (3 x 20 mL), dried (Mg804), 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 wasadded, and the reaction was allowed to come to room
`
`temperature overnight. The mixture was added to saturated NH.C! (15 mL), extracted
`
`with ethyl acetate (3 x 20 mL), dried (MgSOl). filtered. concentrated. and
`
`chromatographed on a 14 cm tall x 26 mm diameter silica gel column eluting with 3:2
`
`hexane:ethyl acetate to afford 18 (147 mg, 52%).
`
`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 acetatethexane to straight ethyl
`
`acetate gradient to afford rv (92 trig;.73%). l’c NMR (coca) (partial spectrum) 5 91.44
`(d, J = 172 Hz, CH). 78.60
`73.54 (CH). 69.97 ((1. J = 23 Hz, CH1), 67.64 (CH).
`
`52.49 (CH), 51.70 (Cl-1). 42.66 (CH2), 34.01 (CH1). 30.01 (d, J = 21 Hz. CH2). 28.84
`
`(CH1), 28.77 (CH1). 26.70 (d. J = 9 Hz. CH2), 24.89 (CH1), 21.79 (CH3). MS, mlz calod.
`
`for CzennosFa [(M + HY], 505.25706; found, 505.25705.
`
`The 15-fluoro PGF's of the present invention may be formulated in various
`
`pharmaceutical compositions for administering to humans and other animak as a treatment
`
`of glaucoma or ocular hypertension. As used herein. the term "pharmaceutically effective
`
`amount” refers to that amount of a compound of the present invention which lowers IOP
`
`
`
`
`"t's
`‘5
`.56'
`2
`a,“
`‘'51:.—
`s":
`
`a {
`
`
`
`thLLASD52.1v1“‘
`
`
`
`- To a suspension of (rt-carboxybutyl)triphenylphosphonium bromide (750 mg. 1.7
`mmol) in THF (5 mL) at 0 °C (bath temperature) was added a l M solution of potassium :-
`butoxide in THF (3.0 ml... 3.0 mmol). After 10 min. a solution of 17 (220 mg) in THF (3
`
`' M
`
`h *
`
`.
`«*5I
`
`.1.
`
`Micro Labs Exhibit 1013-19
`
`

`

`
`
`
`
`aultimo-r"~
`..4w...
`
`when administered to a patient, especially a mammal. The preferred route of administration
`
`.
`
`is topical. _The compounds of the present invention can be administered as solutions.
`
`~ suspensions. or emulsions (dispersions) in an ophthalmieally acceptable vehicle. As used
`the term “ophthalmically acceptable vehicle” refers to any substance or combination
`of substances which are non-reactive with the compounds and suitable for administration to
`
`a patient. Solubilizers and stabilizers are deemed to be non-reactive. Preferred are aqueous
`
`vehicles suitable for topical application to the patient’s eyes.
`
`In fanning composiiions for topical administration, the compounds of the present
`invention are generally ionnulated as between about 0.00003 to about 0.5 percent by
`
`weight (W1%) solutions in water at a pH between 4.5 to 8.0.
`
`'lhe compounds are
`
`preferably formulated as between about 0.0005 to about 0.03 wt% and. most preferably.
`
`between about 0.001 and about 0.01 M95. While the precise regimen is left to the
`
`discretion of the clinician. it is recommended that the resulting solution he topically applied
`
`by placing one drop in each eye one or two times a day.
`
`.17.
`
`Other ingredients which may be desirable to use in the ophthalmic preparations of
`
`the present invention include preservatives. co—solvents and viscosity building agents.
`
`5
`
`.
`
`.
`
`l
`
`. IE
`
`.
`
`:
`
`Ophthalmic productsare typically packaged in multidose form. Preservatives are
`
`thus required to prevent microbial contamination during use. Suitable preservatives
`
`include: bemalkonium chloride. tbimerosal. 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 level
`
`between about 0.001% and about 1.0% by weight.
`
`CttSDlxcnts:
`
`Prostaglandins. and particularly ester derivatives thereof. typically have limited
`
`so
`
`solubility in water and therefore may require a surfactant or other appropriate co-solvcnt in
`
`the composition. Such co-solvents include: Polysorbate 20, 60 and 80; Plumnic F458, F-84
`
`..up»
`
`Micro Labs Exhibit 1013-20
`
`Micro Labs Exhibit 1013-20
`
`

`

`
`
`-—-'
`
`“we.”
`
`j.;
`
`- V.
`
`r"
`
`‘
`
`v
`
`.
`
`
`
`1
`
`i
`
`l
`
`
`
`www"’.‘fl'‘9‘,"3"‘'
`
`
`,
`ii?-
`.-
`taxi
`gkiticl
`Mai:
`
`Ingredient“. "
`fi"""
`
`_
`
`Compound ll
`
`HtosphateBufiered Saline
`
`‘
`
`Polysorbate so
`
`hmunflwflo)
`‘
`'
`
`~
`
`0.005
`
`1.0
`
`as
`
`Pin-itiedwatu
`
`q.s.tolOO%
`
`_.
`
`Micro Labs Exhibit 1013-21
`
`
`
`: fi' .
`
`2
`
`‘.
`
`3
`
`and P-103; CREMOPHORE‘ EL (polyoxyl 35 castor oil) cyclodextrin; or other agents
`
`known to those skilled in the art. Such co-solvents are typically employed at a level
`
`- between about 0.01% and about 2% by weight.
`
`5 museum:
`
`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 decreasephysical separation of components of a suspension or emulsion of
`
`formulation and/or otherylise to improve the ophthalmic formulation. Such viscosity
`building agents include, for example, polyvinyl alcohol, polyvinyl pynolidone. methyl
`cellulose, hydroxy propyl methylcellulose, bydroxyethyl cellulose, carboxymethyl cellulose,
`
`hydroxy propyl cellulose, chondruitin sulfate and salts thereof, hyaluronic acid and salts
`
`thereof, and other agents known to those skilled in the an. Such agents are typically
`
`employed at a level between about 0.01% and about 2% by weight.
`
`Preferred formulations of substituted tetrahydmfurans of the present invention
`include the following Examples 3-6:
`
`in
`
`u
`
`
`
`EXAMPLE8
`
`20
`
`25
`
`
`
`
`Micro Labs Exhibit 1013-21
`
`

`

`
`
`
`
`
`
`
`
`
`
`
`
`'32;av}:.—~:.v.:-,
`
`{I
`’
`"4
`
`L;
`
`EN—
`Ingredient
`
`Ms“—
`COW [I
`0.”
`
`Monowa sodium phospmg
`
`005
`
`Dibasic sodium phosphate
`(anhydmus) .
`‘ "
`
`Sodium ddodde
`
`.'
`
`z.
`
`’-
`
`Disodium EDTAmasodim)
`
`Bennlknnium chloride
`
`HCI and/or NaOH
`
`Purified water
`
`_—
`lugnsdlen!’ W3,
`MN
`Compound m
`0'0“
`0.05
`Mombaslc sodium mm
`
`Dibnic sodium phosphate
`(anhydrous)
`
`KC! and/or mm!
`
`Pulsed was
`
`.-
`
`-
`
`.. «.z
`
`y "
`
`A
`
`_
`
`-
`
`Micro Labs Exhibit 1013-22
`
` -vaum-www-
`
`
`
`
`
`-»--v-r.—.-rV
`
`Micro Labs Exhibit 1013-22
`
`

`

`
`
`
`
`15
`
`'L
`
`
`he 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.
`
`Phosphate Buffered Saline
`
`Hydroxypropyl-B-cydodextrin
`
`mm’Qém '
`
`N T
`
`Micro Labs Exhibit 1013-23
`
`Micro Labs Exhibit 1013-23
`
`

`

`-21-
`
`R = H or cationic salt moiety. or COZR = ophthalmically acceptable ester
`moiety; R‘. R’ = same or difl'ucnt = H or alkyl; R‘ = H, acyl. or alkyl; R’,
`R' = same or different: H. acyl. or alkyl; with the proviso that if one ofR’.
`R' = may]. then the other: H or alkyl;
`
`R’, R’ =_ same or difietent = H. alkyl. or any];
`
`D, D' = difi'crent = H and fluorine;
`
`x=(cnz)q or (CHan whereq = l-6;and
`
`.
`
`o
`whaem:
`
`R‘ = com, CONR‘R’. cmoa‘, or cnzm’k‘; where
`
`n=Oor2;
`
`~— = single or non-cumuiated double bond. with the provisos that a double bond
`between carbons 4 and 5 may not be of the tram configuration; and that a double
`bond between carbons 13 and I4 rnay not be of the cis configuration;
`
`Micro Labs Exhibit 1013-24
`
`

`

`
`
`y
`
`g;
`
`’I“
`
`
`
`i‘m’fi‘arxrrr"flz'v'v-"
`
`
`
`
`
`Y = a phenyl ring optionally substituted with alkyl. halo. tdhalomcthyl, alkoxy,
`
`acyl, acyloxy, amino, alkylamino, acylamino. or bey; or
`
`X‘Y = (CH2)le; where p = 0-6; and
`
`Yr:
`
`W / '
`
`\\ \ 'T_Z or W
`
`whaein:
`
`w = CH1, 0, 3(0)... NR