USOO6399593B1 (12) United States Patent (10) Patent No.: US 6,399,593 B1 Grubb et al. (45) Date of Patent: Jun. 4, 2002

(54) CYCLIC REGIMENS USING CYCLIC UREA 5,733,902 A 3/1998 Schneider ...... 514/177 AND CYCLIC AMIDE DERVATIVES 5,808,139 A 9/1998 Pathirana ...... 560/138 5,874,430 A 2/1999 Christ ...... 514/229.8 (75) Inventors: Gary S. Grubb, Newtown Square; 6,077,840 A 6/2000 Kurihara ...... 514/232.8 Puwen Zhang, Audubon; Arthur A. FOREIGN PATENT DOCUMENTS Santilli, Havertwon; Andrew Fensome, DE 363.3861 4/1988 Wayne; Eugene A. Terefenko, DE 4330 234 3/1995 Quakertown; Andrew Q. Viet, Upper Darby, all of PA (US); Jay E. Wrobel, (List continued on next page.) Lawrenceville, NJ (US); James P. OTHER PUBLICATIONS Edwards, San Diego, CA (US); Todd R.M. Evans, “The and Thyroid Hormone Receptor K. Jones, Solana Beach, CA (US); Superfamily”, Science, 240:889 (May 13, 1988). Christopher M. Tegley, Thousand A. Ulmann et al., “Clinical Uses of (MFP)”, Oaks, CA (US); Lin Zhi, San Diego, Ann. N.Y. Acad. Sci., 261:248 (Jun. 12, 1995). CA (US) R. Kekkonen et al., “Sequential Regiment of the Antiproges terone RU486 and Synthetic Progestin for Contraception”, (73) Assignees: Wyeth, Madison, NJ (US); Ligand Fertility and Sterility, 60(4):610 (Oct. 1993). Pharmaceuticals, Inc., San Diego, CA K. Horwitz et al., “Progestin, Receptors, and (US) Breast Cancer”, “Horm. Cancer', publisher: Birkhaeuser, Boston, Mass., ed. Vedeckis, p. 283-306 (1996) abstract (*) Notice: Subject to any disclaimer, the term of this only. patent is extended or adjusted under 35 A. A. Murphy et al., “Regression of Uterine Leiomyomata in U.S.C. 154(b) by 0 days. Response to the Antiprogesterone RU486”, J. Clin. Endo. Metab., 76(2):513 (Feb. 1993). (21) Appl. No.: 09/552,037 L. M. Kettel et al., “Endocrine Responses to Long-Term (22) Filed: Apr. 19, 2000 Administration of the Antiprogesterone RU486 in Patients with Pelvic Endometriosis”, Fertility and Sterility, Related U.S. Application Data 56(3):402 (Sep.1991). (60) Provisional application No. 60/198.238, filed on May 4, H. Michna et al., “Differentiation Therapy with Progesterone 1999. Antagonists”, Ann, N.Y. Acad. Sci., 761:224 (Jun. 1995). (51) Int. Cl...... A61K 31156; A61K 31/535; (List continued on next page.) A61K 31/44 Primary Examiner Minna Moezie (52) U.S. Cl...... 514/171; 514/230.5; 514/224.5; Assistant Examiner San-Ming Hui 514/229.2; 514/293; 514/300 (74) Attorney, Agent, or Firm Howson and Howson (58) Field of Search ...... 514/171,230.5, (57) ABSTRACT 514/224.5, 229.2, 293,300 This invention concerns cyclic combination therapies using (56) References Cited progestational agents and indoline derivatives which are antagonists of the general Structure: U.S. PATENT DOCUMENTS 3,635,964 A 1/1972 Skorcz et al...... 260/247.1 I 3,917,592 A 11/1975 Kobzina ...... 260/244 R1 R2 4,093,730 A 6/1978 Butti ...... 424/270 4,440,785 A 4/1984 Walsh ...... 424/317 R A. 4,666,913 A 5/1987 Kubla et al...... 514/259 y n W 4,670,566 A 6/1987 Walsh ...... 548/485 4,721,721 A 1/1988 Kuha ...... 514/312 B n 4. N1. O 4,822,794. A 4/1989 Spada ...... 514/230 4,831,027 A 5/1989 Narr et al...... 514/212 4.853,473 A 8/1989 Fischer et al. . ... 549/326 k 5,007,952 A 4/1991 Kume et al...... 71/73 5,171,851 A 12/1992 Kim et al...... 544/50 wherein: A, B and D are N or CH, though not all can be CH; 5,414,088 A 5/1995 Von Der Saal et al...... 546/158 R" and R are H, COR, NRCOR, or optionally substi 5,453,516 A 9/1995 Fischer et al...... 54.8/543 tuted alkyl, alkenyl, alkynyl, cycloalkyl, or aryl groups; or 5,475,020 A 12/1995 Johnson et al...... 548/466 R" and R form a spirocyclic ring; R is H or optionally

5,521,166 A 5/1996 Grubb ...... 514/170 5,681,817 A 10/1997 Hodgen et al...... 514/12 substituted alkyl aryl, alkoxy, or aminoalkyl; R is H, alkyl 5,688.808 A 11/1997 Jones et al...... 514/285 alkyl; R is H, OH, NH, alkyl, alkenyl, or COR; R is H, 5,688.810 A 11/1997 Jones et al...... 514/311 alkyl, aryl, alkoxy, or aminoalkyl, R' is benzene or a 5 or 6 5,693,646 A 12/1997 Jones et al...... 514/285 membered heteroaromatic ring; R is H, alkyl, aryl, alkoxy, 5,693,647 A 12/1997 Jones et al...... 514/285 or aminoalkyl; R is H or alkyl; R is H or alkyl; W is O or 5,696,127 A 12/1997 Jones et al...... 514/285 a chemical bond; or a pharmaceutically acceptable Salt 5,696,130 A 12/1997 Jones et al...... 514/291 thereof. These methods may be used for contraception. 5,696,133 A 12/1997 Pooley et al...... 514/314 5,719,136 A 2/1998 Chwalisz et al...... 514/170 20 Claims, No Drawings US 6,399,593 B1 Page 2

FOREIGN PATENT DOCUMENTS A. Andreani et al., “Potential Antitumor Agents XVII (1). DE 43 44 463 6/1995 Cytotoxic Agents from Indole Derivatives and Their Inter EP O22.317 1/1981 mediates”, Acta. Pharm. Nord, 2(6):407 (1990). EP O 208 510 1/1987 Sakata et al., “Silver Halide Photographic Materials Useful EP 31.1135 4/1989 for Platemaking", Chemical Abstracts, 123:30.1431 (1993). EP 385850 9/1990 P. Pflegel et al., “Polarografie CO EP 483.077 9/1991 7-Chlor-5-phenyl-2-thioxo-1H-2,3-dihydro-1,3,4-ben EP 454330 10/1991 zotriazepinen”, Pharmazie, 37(10): 714-717 (1982). EP OS35529 9/1992 E. I. Barengolts et al., “Progesterone Anatagonist RU 486 EP 510235 10/1992 Has Bone-Sparing Effects in Ovariectomized Rats', Bone, EP 947 SO7 10/1999 EP 978 279 2/2000 17(1):21 (Jul 1995). JP 63112584 5/1988 E. V. Gromachevskaya et al., “Studies of 4H-3, 1-Benzox WO WO 86/03749 7/1986 azines”, Chem. Heterocycl. Cmpds. 33(10): 1209–1214 WO WO 91/04974 4/1991 (1997). WO WO 91/06545 5/1991 D. Chiarino et al., “2, 1-Benzisothiazoline 2, 2-Dioxide and WO WO 93/12085 6/1993 Derivatives”, J. Heterocycl. Chem., 23(6):1645-1649 WO WO 94/14434 7/1994 (Nov.-Dec. 1986). WO WO 94/29272 12/1994 A. Turck et al., “On the Metabolism of 3-Substituted and 3, WO WO95/11013 4/1995 WO WO95/20389 8/1995 6-Disubstituted Pyridazines”, Tetrahedron, 49(3):599–606 WO WO95/20972 8/1995 (1993). WO WO95/33746 12/1995 V. Kumar et al., “Synthesis of 7-AZaindole and 7-AZaox WO WO 96/15794 5/1996 indole Derivatives through a Palladium-Catalyzed Cross WO WO 96/19458 6/1996 Coupling Reaction”, J. Org. Chem., 57(25):6995-6998 WO WO 96/19997 7/1996 (1992). WO WO 97/13767 4/1997 P. Canonne et al., “Spirocyclization of 1-(o-Aminophenyl WO WO 97/49.407 12/1997 )cycloalkanols and 1-(2-Amino-3-pyridinyl) cycloal WO WO 98/14436 4/1998 WO WO 98/27059 6/1998 kanols”, J. Heterocyclic Chem., 26:113 (Jan.-Feb. 1989). WO WO 98/55116 12/1998 M-C. Forest et al., “A Novel Class of Cardiotonic Agents: WO WO 99/10325 3/1999 Synthesis and Biological Evaluation of 5-Substituted WO WO 99/11264 3/1999 3,6-Dihydrothiadiazin-2-ones with Cyclic AMP Phos WO WO 99/15500 4/1999 phodiesterase Inhibiting and Myofibrillar Sensitiz WO WO 99/44608 9/1999 ing Properties”, J. Med. Chem., 35:163–172 (Jan. 1992). D. W. Combs et al., “Heteroatom Analogues of Bemoradan: OTHER PUBLICATIONS Chemistry and Cardiotonic Activity of 1,4-Benzothiazi nylpyridaziones”, J. Med. Chem., 35:172-176 (Jan. 1992). L. Zhi et al., “5-Aryl-1,2-Dihydrochromeno 3,4-fcuino Kurihari et al., “Synthesis of (+)-PF1092A, B, and C; New lines: A Novel Class of Human Progesterone Nonsteroidal Progesterone Receptor Ligands”, J. Antibiot Receptor Agonists”, J. Med Chem., 41(3):291 (Oct. 22, ics, 50(4):360 (Apr. 1997). 1998). A. Kende et al., “Regioselective C-3 Alkylation of Oxindole D. W. Combs et al., “Nonsteroidal Progesterone Receptor Dianion”, Synth. Commun. 12(1):1 (1982). Ligands. 2. High-Affinity Ligands with Selectivity for Bone T. Tucker et al., “Synthesis of a Series of Cell Progesterone Receptors”, J. Med. Chem., 38:4880 (Dec. 4-(Arylethylnyl)-6-Chloro-4-Cyclopropyl-3, 8, 1995). 4-dihydroquinazolin-2(1H)-ones as Novel Non-Nucleo K. L. Perlman et al., “20-Oxopregnacalciferols: Vitamin D side HIV-1 Reverse Transcriptase Inhibitors”, J. Med. Compounds that Bind the Progesterone Receptor”, Tet. Chem., 37:2347–2444 (Jul 22, 1994). Letters, 35(15):2295 (1994). J. P. Edwards et al., “5-Aryl-1,2-Dihydro-5H-Chromeno L. G. Hamann et al., “Synthesis and Biological Activity of 3,4-fCuinolines as Potent, Orally Active, Nonsteroidal Novel Nonsteroidal Progesterone Receptor Antagonists”, Progesterone Receptor Agonists: The Effect of D-Ring Ann. N. Y. Acad. Sci., 761:383 (Jun. 12, 1995). Substituents”, J. Med. Chem., 41:303-310 (Jan. 29, 1998). R. H. K. Chen et al., “Synthesis and SAR of a Novel Series Derwent WPI abstract, “New Imidazo-Pyridine of Spirobenzothlzaepine Derivatives with Antiprogestin Derivatives - Useful as Platelet Agglutination Inhibitor, Antiallergic, Antiinflammatory Sedative, Cardiac, and Car Activity", POI-37, 16" Int. Cong. Het. Chem., Montana diovascular Vasodilators”, JP 63112584 (May 1988). (1997). Derwent WPI abstract, N. Brumagniez et al., “Benzimida B. Narr et al., “Preparation, Testing, and Formulation of Zole and AZabenzimidazole(s)-Having Cardiotonic, Imidazobenzoxazinones as Cardiotonics', Chemical Vasodilating, Anti-Hypertensive, Anti-Aggregation, and Abstracts, 109:22973 (1988). Anti-Ulcer Activity”, EP385850 (Sep. 1990). R. J. Hartmann et al., “Effects of Brofoxine, A New Anxi Derwent WPI abstract, F. Arndt et al., “New Heterocycle olytic on Experimentally Induced Conflict in Rats', Proc Substituted Benzo-Fused Azine and Azole Derivatives - West. Pharmacol. Soc., 21:51-55 (1978). Useful as Selective Herbicides for Pre or Post-Emergence B. Singh et al., “Novel cAMP PDE III Inhibitor: Imidazo[4, Application”, EP31 1135 (Apr. 1989). 5-b]pyridin-2(3H)-ones and Thiazolo 4,5-b] Vinod K. Jhalani et al., “A Convenient Synthesis of pyridin-2(3H)-ones and Their Analogs”, J. Med. Chem., 5-Keto-6-methyl-5,6-dihydro-1,6-naphthyridine”, Indian 37:248 (Jan. 21, 1994). Journal of Chemistry, 22B:916 (Sep. 1983). US 6,399,593 B1 Page 3

K. Horwitz et al., “Progestin, Progesterone Receptors, and tives of 2-Benzoxazolone.” Chemical Abstracts, vol. 99, Breast Cancer”, “Hormones and Cancer', publisher: No. 1, Abst. No. 157a, Jul. 4, 1983. Birkhaeuser, Boston, Mass., ed. Vedeckis, p. 283-306 (1996). Meanwell N.A., et al. “Regiospecific Functionalization of Mamaev, V.P., et al., “Synthesis of 4H-Thieno 3.2-B 1,3-dihydro-2H-Benzimidazol-2-One and Structurally Pyrrol-5(6H)-One” Bulletin of the Academy of Sciences on Related Cyclic Urea Derivatives”. J. Organic Chem., 60(6): the USSR. Division of Chemical Science US Consultants 1565–82 (Mar. 24, 1995). Bureau. New York. Vol. 9, p. 1549-1553, 1966. Singh, B., et al., “An Efficient and Novel Synthesis of Fused Derwent WPI Abstract, Chwalisz, K., et al. “Female Con traceptive Method Comprises Gestation Treatment with Thiazol-2(3H)-ones” Heterocycles, 36(1): 133-134, p. 136, Intermittent Progesterone Antagonist Administration., DE compounds 16a, 18a, Jan. 1993. 4,330,234. (Mar. 1995). Vernin, G., et al., “Etude Dans la Serie des Radicaux Derwent WPI Abstract, Chwalisz, K., et al. “Contraceptive Heterocycliques. Partie XV. Decomposition aprotique de 1 Pack for Implantation Inhibition -Contains Competitive amino-6-ethyl-2-benzothiazole dans des Substrats aroma Progesterone Antagonist and Gestagen for Sequential Oral tiques et heteroaromatiques: preparation des mesityl-6- et Administration.”, DE 4,344,463. (Jun. 1995). furyl-6-ethyl-2-benzothiazoles, des Sels quaternaires et des Kolasa, K., et al., “Preliminary Pharmacological Studies of Spiropyrannes correspondants' Helvetica Chimica Acta, the Central Action of Phenyl and Piperidinomethyl Deriva 62(1/3):21–30 Jan. 24, 1979. US 6,399,593 B1 1 2 CYCLIC REGIMENS USING CYCLIC UREA tone and , have been shown to be effective in a AND CYCLIC AMIDE DERVATIVES model of hormone dependent prostate cancer, which may indicate their utility in the treatment of this condition in men CROSS-REFERENCE TO RELATED APPLICATIONS (Michna, et al, Ann. N.Y. Acad. Sci., 761, 224, 1995). This application claims the benefit of the priority of U.S. Provisional Patent Application No. 60/198,238, filed May 4, Jones, et al., (U.S. Pat. No. 5,688,810) describe the PR 1999. antagonist dihydroquinoline 1. FIELD OF THE INVENTION This invention relates to regimens of administering com N pounds which are antagonists of the progesterone receptor in W combination with a progestin, an estrogen, or both. 15 s Me BACKGROUND OF THE INVENTION S Intracellular receptors (IR) form a class of structurally 2 N related gene regulators known as “ligand dependent tran Me scription factors” (R. M. Evans, Science, 240, 889, 1988). N The steroid receptor family is a subset of the IR family, H Me including progesterone receptor (PR), estrogen receptor (ER), receptor (AR), glucocorticoid receptor (GR), and mineralocorticoid receptor (MR). 25 The natural hormone, or ligand, for the PR is the steroid Jones, et al, described the enol ether 2 (U.S. Pat. No. progesterone, but Synthetic compounds, Such as medroX 5,693,646) as a PR ligand. yprogesterone or , have been made which also serve as ligands. Once a ligand is present in the fluid Surrounding a cell, it passes through the membrane Via Me passive diffusion, and binds to the IR to create a receptor/ ligand complex. This complex binds to specific gene pro moters present in the cells DNA. Once bound to the DNA the complex modulates the production of mRNA and protein 35 encoded by that gene. A compound that binds to an IR and mimics the action of the natural hormone is termed an agonist, whilst a compound which inhibits the effect of the hormone is an antagonist. PR agonists (natural and Synthetic) are known to play an impor 40 tant role in the health of women. PR agonists are used in birth control formulations, typically in the presence of an ER agonist. ER agonists are used to treat the Symptoms of menopause, but have been associated with a proliferative 45 effect on the uterus which can lead to an increased risk of Jones, et al, described compound 3 (U.S. Pat. No. 5,696, uterine cancers. Co-administration of a PR agonist reduceS/ 127) as a PR ligand. ablates that risk. PR antagonists may also be used in contraception. In this 3 context they may be administered alone (Ulmann, et al., Ann. 50 Me Cl N.Y. Acad. Sci., 261,248, 1995), in combination with a PR agonist (Kekkonen, et al, Fertility and Sterility, 60, 610, 1993) or in combination with a partial ER antagonist such as O ( ) tamoxifen (WO 96/19997 A1, Jul. 4, 1996). PR antagonists 55 may also be useful for the treatment of hormone dependent Me breast cancers (Horwitz, et al, Horm. Cancer, 283, pub: F N Birkhaeuser, Boston, Mass., ed. Vedeckis) as well as uterine Me and ovarian cancers. PR antagonists may also be useful for 60 N Me the treatment of non-malignant chronic conditions Such as H fibroids (Murphy, etal, J. Clin. Endo. Metab., 76,513, 1993) and endometriosis (Kettel, et al, Fertility and Sterility, 56, 402, 1991). PR antagonists may also be useful in hormone replacement therapy for post menopausal patients in com 65 bination with a partial ER antagonist Such as tamoxifen Zhi, et al, described lactones 4, 5 and 6 as PR antagonists (U.S. Pat. No. 5719136). PR antagonists, such as mifepris (J. Med. Chem, 41,291, 1998). US 6,399,593 B1

4

CH CH

5 O CH2 O Me

15 N HO s Me

N H Me Hamann, et al, described the PR antagonist 10 (Ann, N.Y. 6

Acad. Sci., 761, 383, 1995).

1O OMe

Br CH2 Me 25

OAc Me Me

Zhi, etal, described the ether 7 as a PR antagonist (J. Med. Chen, et al, described the PR antagonist 11 (Chen, et al., Chem., 41,291, 1998). POI-37, 16'. Int. Cong. Het. Chem., Montana, 1997).

7 35 11 Cl

C 40 O Me

N Me 45 N H Me cy

50 Kurihari, et. al., described the PR ligand 12 (J. Antibiotics, 50, 360, 1997). Combs, et al., disclosed the amide 8 as a ligand for the PR (J. Med. Chem., 38, 4880, 1995). 12

55 OAc

N Br 60 The patent by Christ et al. (WO 9814436) claims cyclo amide such as compound A as inhibitors of HIV reverse transcriptase. Other prior art includes pyridazine cyclo 65 amide such as compound B by Turck et al. (Tetrahedron, Perlman, et. al., described the vitamin D analog 9 as a PR 49(3), 599–606(1993)) and compound such as C by ligand (Tet. Letters, 35, 2295, 1994). Canonne et al. (J. heterocyclic Chem. 26, 113(1989)). No US 6,399,593 B1 S 6 activity data were reported in Turek's and Canonne publi with one or more progestational agents. This invention cations. further provides methods of treatment and dosing regimens further utilizing in combination with these antiprogestins A.

and progestins, an estrogen, Such as ethinyl . These regimens and combinations may be administered to a mammal to induce contraception or for the treatment and/or prevention of Secondary amenorrhea, dysfunctional bleeding, uterine leiomyomata, endometriosis, polycystic ovary Syndrome, carcinomas and adenocarcinomas of the endometrium, ovary, breast, colon, prostate. Additional uses of the invention include stimulation of food intake. The uses herein for the treatment and/or prevention of the conditions or diseases described above includes the continuous admin 15 istration or periodic discontinuation of administration of the B invention to allow for minimization of effect dose or mini mization of Side effects or cyclic menstrual bleeding. The use of this invention for contraception includes administration, preferably orally, to a female of child bear ing age an antiprogestin in combination with an estrogen or 21 O progestin or both. These administration regimens are pref erably carried out over 28 consecutive days, with a terminal portion of the cycle containing administration of no NinN N 1sO 25 H progestins, estrogens or anti-progestins. C The progestins of these combinations may be adminis tered alone or in combination with an estrogen for the first 14-24 days of the cycle, the progestins being administered 21 O at a dosage range equal in progestational activity to about 35 tug to about 150 lug levonorgestrel per day, preferably equal s-s-sN N O in activity to from about 35 lug to about 100 ug levonorg H estrel per day. An antiprogestin may then be administered 35 alone or in combination with an estrogen for a period of 1 Regarding cyclic amides, Singh et al. and Kumar et al. to 11 days to begin on any cycle day between day 14 and 24. (Singh et al. J. Med. Chem. 37(2), 248-254(1994); Kumaret The anti-progestin in these combinations may be adminis al. J. Org. Chem. 57(25), 6995-6998(1992)) disclose com tered at a dose of from about 2 ug to about 50 ug per day and pounds such as D and E claimed as cAMP PDE III inhibi the estrogen may be administered at a dose of from about 10 torS. 40 tug to about 35 lug per day. In an oral administration, a package or kit containing 28 tablets will include a placebo

tablet on those days when the antiprogestin or progestin or estrogen is not administered. 45 In a preferred embodiment of this invention, the progestins of this invention may be administered alone or in combination with estrogen for the initial 18 to 21 days of a 28-day cycle, followed by administration of an antiprogestin, alone or in combination with an estrogen, for 50 from 1 to 7 days. The estrogen to be used in the combinations and formu lations of this invention is preferably ethinyl estradiol. ProgeStational agents useful with this invention include, 55 but are not limited to, levonorgestrel, , , 3-ketodesogestrel, norethindrone, , norethindrone acetate, , OSaterone, acetate, , , droSpire none, , or (17 U.S Pat. No. 5,521,166 (Grubb) teaches cyclophasic deacetyl)norgestimate. Among the preferred progestins for hormonal regimens comprising an antiprogestin and a 60 progestin wherein the progestin is administered in the alter use in the combinations of this invention are levonorgestrel, nating presence and absence of an antiprogestin. The dis gestodene and trimegestone. closed regimens also provide for use of an estrogen for a Examples of orally administered regimens of this inven period of from 2–4 days to prevent breakthrough bleeding. tion over a 28 day cycle include administration of a proges DESCRIPTION OF THE INVENTION 65 tational agent Solely for the first 21 days at a daily dose equal This invention provides combination therapies and dosing in progestational activity to from about 35 to about 100 lug regimens utilizing antiprogestational agents in combination of levonorgestrel. An antiprogestin compound of this inven US 6,399,593 B1 7 8 tion may then be administered at a daily dose of from about daily dosage unit containing an antiprogestin com 2 to 50 mg from day 22 to day 24, followed by no pound at a daily dosage of from about 2 to 50 mg, and administration or administration of a placebo for days 25 to c) optionally, a third phase of 4 daily units of an orally and 28. It is most preferred that the daily dosages of each pharmaceutically acceptable placebo for each of days relevant active ingredient be incorporated into a combined, 5 25 to 28. Single daily dosage unit, totaling 28 daily units per 28-day Another 28-day cycle packaging regimen or kit of this cycle. invention comprises: In another regimen, a progestational agent may be coad a) a first phase of from 18 to 21 daily dosage units of a ministered for the first 21 days at a daily dose equal in progestational agent equal in progestational activity to progestational activity to from about 35 to about 150 ug about 35 to about 150 ug levonorgestrel, preferably levonorgestrel, preferably equal in activity to from about 35 equal in activity to from about 35 to about 100 tug to about 100 lug levonorgestrel, with an estrogen, Such as levonorgestrel, and, as an estrogen, ethinyl estradiol at ethinyl estradiol, at a daily dose range of from about 10 to a daily dose range of from about 10 to about 35 lig; and about 35 lug. This may be followed as described above with 15 b) a second phase of from 1 to 7 daily dosage units of an an antiprogestin administered at a daily dose of from about antiprogestin of this invention at a daily dose of from 2 to 50 mg from day 22 to day 24, followed by no about 2 to 50 mg; and administration or administration of a placebo for days 25 to c) optionally, an orally and pharmaceutically acceptable placebo for each of the remaining 0-9 days in the 28. 28-day cycle in which no progestational agent, estrogen Still another regimen within the scope of this invention or antiprogestin is administered. will include coadministration from dayS 1 to 21 of a proges A preferred embodiment of the kit described above may tational agent, the progestational agent, preferably comprise: levonorgestrel, being administered at a daily dose equal in progestational activity to from about 35 to about 100 ug a) a first phase of 21 daily dosage units of a progestational 25 agent equal in progestational activity to about 35 to levonorgestrel, and an estrogen, Such as ethinyl estradiol at about 150 lug levonorgestrel, preferably equal in activ a daily dose range of from about 10 to about 35ug. This will ity to from about 35 to about 100 lug levonorgestrel, be followed on days 22 to 24 by coadministration of an and, as an estrogen, ethinyl estradiol at a daily dose antiprogestin (2 to 50 mg/day) and an estrogen, Such as range of from about 10 to about 35 lug; and ethinyl estradiol, at a daily dose of from about 10 to about b) a second phase of 3 daily dosage units for days 22 to 35 ug. From day 25 to day 28, this regimen may be followed 24 of an antiprogestin administered at a daily dose of by no administration or administration of a placebo. from about 2 to 50 mg; and This invention also kits or packages of pharmaceutical c) optionally, a third phase of 4 daily dose units of an formulations designed for use in the regimens described orally and pharmaceutically acceptable placebo for herein. These kits are preferably designed for daily oral 35 each of days 25 to 28. administration over a 28-day cycle, preferably for one oral A further 28-day packaged regimen or kit of this invention administration per day, and organized So as to indicate a comprises: Single oral formulation or combination of oral formulations a) a first phase of from 18 to 21 daily dosage units, each to be taken on each day of the 28-day cycle. Preferably each containing a progestational agent of this invention at a 40 daily dose equal in progestational activity to about 35 kit will include oral tablets to be taken on each the days to about 150 lug levonorgestrel, preferably equal in Specified, preferably one oral tablet will contain each of the activity to from about 35 to about 100 lug combined daily dosages indicated. levonorgestrel, and ethinyl estradiol at a daily dose According to the regimens described above, one 28-day range of from about 10 to about 35 lug; kit may comprise: 45 b) a second phase of from 1 to 7 daily dose units, each a) an initial phase of from 14 to 21 daily dosage units of daily dose unit containing an antiprogestin of this a progestational agent equal in progestational activity invention at a concentration of from 2 to 50 mg, and to about 35 to about 150 lug levonorgestrel, preferably ethinyl estradiol at a concentration of from about 10 to equal in progestational activity to about 35 to about 100 about 35 lug; and pig levonorgestrel; 50 c) optionally, an orally and pharmaceutically acceptable b) a second phase of from 1 to 11 daily dosage units of an placebo for each of the remaining 0-9 days in the antiprogestin compound of this invention, each daily 28-day cycle in which no progestational agent, estrogen dosage unit containing an antiprogestin compound at a or antiprogestin is administered. daily dosage of from about 2 to 50 mg, and 55 A preferred embodiment of the package or kit just c) optionally, a third phase of an orally and pharmaceu described comprises: tically acceptable placebo for the remaining days of the a) a first phase of 21 daily dosage units, each containing cycle in which no antiprogestin, progestin or estrogen a progestational agent of this invention at a daily dose is administered. equal in progestational activity to about 35 to about 150 A preferred embodiment of this kit may comprise: 60 tug levonorgestrel, preferably from about 35 to about a) an initial phase of 21 daily dosage units of a proges 100 ug levonorgestrel, and ethinyl estradiol at a daily tational agent equal in progestational activity to about dose range of from about 10 to about 35 lug; 35 to about 150 lug levonorgestrel, preferably equal in b) a second phase of 3 daily dose units for days 22 to 24, progestational activity to about 35 to about 100 ug each dose unit containing an antiprogestin of this levonorgestrel; 65 invention at a concentration of from 2 to 50 mg, and b) a Second phase of 3 daily dosage units for days 22 to ethinyl estradiol at a concentration of from about 10 to 24 of an antiprogestin compound of this invention, each about 35 lug; and US 6,399,593 B1 9 10 c) optionally, a third phase of 4 daily units of an orally and R is H, C, to C, alkyl, substituted C, to C, alkyl, aryl, pharmaceutically acceptable placebo for each of days Substituted aryl, C to C alkoxy, Substituted C to C. 25 to 28. alkoxy, C to C aminoalkyl, or Substituted C to C. In each of the regimens and kits just described, it is aminoalkyl, preferred that the daily dosage of each pharmaceutically R’ is H, C, to C alkyl, or substituted C to C alkyl; active component of the regimen remain fixed in each R is H, OH, NH, C, to C alkyl, Substituted C to C, particular phase in which it is administered. It is also alkyl, C. to Calkenyl, Substituted C to Calkenyl, or understood that the daily dose units described are to be COR; administered in the order described, with the first phase R is H, C, to C alkyl, substituted C to C alkyl, aryl, followed in order by the second and third phases. To help Substituted aryl, C to C alkoxy, Substituted C to C. facilitate compliance with each regimen, it is also preferred alkoxy, C to C aminoalkyl, or Substituted C to C. that the kits contain the placebo described for the final days aminoalkyl, of the cycle. It is further preferred that each package or kit R" is a trisubstituted benzene ring containing the Substitu comprise a pharmaceutically acceptable package having ents X, Y and Z as shown below: indicators for each day of the 28-day cycle, Such as a labeled 15 blister package or dial dispenser packages known in the art. In this disclosure, the terms anti-progestational agents, Z. anti-progestins and progesterone receptor antagonists are understood to be Synonymous. Similarly, progestins, proges e/ N tational agents and progesterone receptor agonists are under stood to refer to compounds of the Same activity. These dosage regimens may be adjusted to provide the optimal therapeutic response. For example, Several divided X is taken from the group including halogen, CN, C to doses of each component may be administered daily or the 25 C alkyl, Substituted C to C alkyl, C to Calkoxy, dose may be proportionally increased or reduced as indi Substituted C to C alkoxy, C to C thioalkoxy, cated by the exigencies of the therapeutic Situation. In the Substituted C to C thioalkoxy, amino, C to C descriptions herein, reference to a daily dosage unit may also aminoalkyl, Substituted C to C aminoalkyl, NO, include divided units which are administered over the course C to C perfluoroalkyl, 5 or 6 membered heterocy of each day of the cycle contemplated. clic ring containing 1 to 3 heteroatoms, CORP, Compounds of this invention which may be used as the OCORP, or NRFCOR; anti-progestational agents in the kits, methods and regimens R’ is H, C, to C alkyl, substituted C, to C, alkyl, aryl, herein are those of the Formula I: substituted aryl, C to Calkoxy, substituted C to C. alkoxy, C to C aminoalkyl, or Substituted C to C. 35 I aminoalkyl, R’ is H, C, to C, alkyl, or substituted C, to C, alkyl; R1 R2 Y and Z are independent substituents taken from the R A. group including H, halogen, CN, NO, C to C n W alkoxy, C to C alkyl, or C to C thioalkoxy; or 40 R" is a five or six membered ring with 1, 2, or 3 B N 2 N 1sO heteroatoms from the group including O, S, SO, SO or NR and containing one or two independent k Substituents from the group including H, halogen, CN, NO and C to C alkyl, C. to C alkoxy, C to wherein: 45 Caminoalkyl, COR, or NRCOR; A, B and D are N or CH, with the proviso that A, B and R is H, C, to C, alkyl, substituted C, to C alkyl, aryl, D can not all be CH; Substituted aryl, C to Calkoxy, Substituted C to C. R" and R are independent substituents selected from H, alkoxy, C to C aminoalkyl, or Substituted C to C. C to C alkyl, Substituted C to C alkyl, C. to C aminoalkyl, alkenyl, Substituted C to Calkenyl, C. to C alkynyl, 50 R’ is H, C, to C, alkyl, or substituted C, to C, alkyl; Substituted C to C alkynyl, C. to Cs cycloalkyl, R is H or C, to C, alkyl; Substituted C to Cs cycloalkyl, aryl, Substituted aryl, W is O or a chemical bond; heterocyclic, substituted heterocyclic, COR, NR’. or a pharmaceutically acceptable Salt thereof. COR; When W is a chemical bond, it is understood that Formula or R' and Rare fused to form a spirocyclic ring selected 55 from a), b) or c), each Spirocyclic ring optionally I exists as: Substituted by from 1 to 3 C-C alkyl groups: a) a 3 to 8 membered Spirocyclic alkyl ring; R1 R2 b) a 3 to 8 membered Spirocyclic alkenyl ring; or R A. c) an optionally substituted 3 to 8 membered hetero 60 n cyclic ring containing one to three heteroatoms from O. the group including O, S and N, the Spirocyclic rings B na N ofa), b) and c) being optionally substituted by from V 1 to 4 groups Selected from fluorine, C to C alkyl, R3 C to C alkoxy, C to C thioalkyl, -CF, -OH, 65 -CN, NH, -NH(C. to C alkyl), or -N(C. to C Preferred antiprogestin compounds are those of the for alkyl). mula: US 6,399,593 B1 11 12 or NR and containing one or two independent R1 R2 Substituents from the group including H, halogen, R A. CN, NO, and C to C alkyl, or C to C, alkoxy; n W R is H or C, to C, alkyl; W is O or a chemical bond; Ba 2 Y; O or a pharmaceutically acceptable Salt thereof. R3 Still, more preferred antiprogestin compounds are those of the formula: wherein: R1 R2 A, B and D are N or CH, with the proviso that A, B and D can not all be CH; R A. R" is H, C, to C alkyl, substituted C to C alkyl, C. to n W Cs cycloalkyl, Substituted C to Cs cycloalkyl, aryl, 15 Ba 2 Substituted aryl, heterocyclic, Substituted heterocyclic, N O COR, or NRCOR; R3 R is H, C, to C, alkyl, substituted C, to C alkyl, C. to C alkenyl, Substituted C to C alkenyl, C. to Cs cycloalkyl, Substituted C to C cycloalkyl, aryl, Sub wherein: Stituted aryl, heterocyclic, Substituted heterocyclic, COR, or NRCOR; or A, B and D are N or CH, with the proviso that A, B and R" and R are fused to form the optionally substituted 3 D cannot all be CH; to 8 membered Spirocyclic alkyl, alkenyl or heterocy 25 R=R and are selected from the group of C to C alkyl, clic rings described above; Substituted C to C alkyl, or Spirocyclic alkyl con R is H, C, to C, alkyl, substituted C, to C, alkyl, aryl, structed by fusing R' and R to form a 3 to 6 membered Substituted aryl, C to C alkoxy, Substituted C to C. Spirocyclic ring; alkoxy, C to C aminoalkyl, or Substituted C to C. R is H, OH, NH, C, to C alkyl, Substituted C to C, aminoalkyl, alkyl, or COR; R is H, C, to C alkyl, or substituted C to C alkyl; R is H, OH, NH, C, to C alkyl, Substituted C to C R is H, C to C alkyl, or C to Calkoxy; alkyl, C. to Calkenyl, Substituted C to Calkenyl, or R" is a disubstituted benzene ring containing the Substitu COR; ents X, and Y as shown below: 35 R is H, C, to C alkyl, Substituted C to C alkyl, aryl, Substituted aryl, C to C alkoxy, Substituted C to C. alkoxy, C to C aminoalkyl, or Substituted C to C. aminoalkyl, R" is a trisubstituted benzene ring containing the Substitu ents X, Y and Z as shown below: 40

rá 45 X is Selected from the group of halogen, CN, C to C. N alkoxy, C to C alkyl, NO, C to C perfluoroalkyl, 5 membered heterocyclic ring containing 1 to 3 heteroatoms, or C to C thioalkoxy, Y is a substituent on the 4 or 5' position from the group X is Selected from halogen, CN, C to C alkyl, 50 Substituted C to C alkyl, C to C alkoxy, Substi including H, halogen, CN, NO, C to C alkoxy, C to tuted C to C alkoxy, C to C thioalkoxy, Substi C alkyl, or C to C thioalkoxy; or tuted C to C thioalkoxy, C to C aminoalkyl, R" is a five membered ring with the structure: Substituted C to C aminoalkyl, NO, C to C. perfluoroalkyl, a 5 membered heterocyclic ring con 55 X taining 1 to 3 heteroatoms, CORP, OCORP, or NRFCORP; pay R’ is H, C to C alkyl, Substituted C to C alkyl, aryl, Y'--UN/ A Substituted aryl, C to C alkoxy, Substituted C to C. alkoxy, C to C aminoalkyl, or Substituted C to C 60 aminoalkyl, R’ is H, C, to C, alkyl, or substituted C, to C, alkyl; U is O, S, or NR; Y and Z are independent substituents selected from H, halogen, CN, NO, C to C alkoxy, C to C alkyl, R is H, or C, to C, alkyl, or C, to C, CO-alkyl; or C to C thioalkoxy; or 65 X is selected from halogen, CN, NO, C, to C alkyl, or R" is a five or six membered ring with 1, 2, or 3 C to C alkoxy, heteroatoms from the group including O, S, SO, SO Y is selected from H and C to C alkyl; or US 6,399,593 B1 13 14 R" is a six membered ring with the structure: istry in Formula I, the present invention includes Such optical isomers and diastereomers, as well as the racemic and resolved, enantiomerically pure R and S Stereoisomers, s as well as other mixtures of the R and S Stereoisomers and NN2 pharmaceutically acceptable Salts thereof. The term “alkyl is used herein to refer to both straight and branched-chain Saturated aliphatic hydrocarbon groups having one to eight carbon atoms, preferably one to Six X is N or CX; carbon atoms; “alkenyl' is intended to include both straight X is halogen, CN or NO; and branched-chain alkyl groups with at least one carbon or a pharmaceutically acceptable Salt thereof. carbon double bond and two to eight carbon atoms, prefer Further preferred antiprogestational compounds include ably two to six carbon atoms; “alkynyl group is intended to those of the formula: cover both Straight- and branched-chain alkyl groups with at 15 least one carbon-carbon triple bond and two to eight carbon atoms, preferably two to Six carbon atoms. R A. The terms “substituted alkyl”, “substituted alkenyl', and n W “substituted alkynyl' refer to alkyl, alkenyl, and alkynyl as just described having one or more Substituents from the Ba 2 group including halogen, CN, OH, NO, amino, aryl, N; O heterocyclic, Substituted aryl, Substituted heterocyclic, alkoxy, aryloxy, Substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, arylthio. These Substituents may wherein: 25 be attached to any carbon of an alkyl, alkenyl, or alkynyl A, B and D are N or CH, with the proviso that A, B and group provided that the attachment constitutes a stable D can not all be CH; chemical moiety. R'=R and are selected from CH and spirocyclic alkyl The term “aryl' is used herein to refer to an aromatic constructed by fusing R' and R to form a 6 membered System which may be a single ring or multiple aromatic rings Spirocyclic ring; fused or linked together as Such that at least one part of the R is H, OH, NH, CH, substituted methyl, or COR; fused or linked rings forms the conjugated aromatic System. R is H, C, to C, alkyl, or C to Calkoxy; The aryl groups include but not limited to phenyl, naphthyl, R" is a disubstituted benzene ring containing the Substitu biphenyl, anthryl, tetrohydronaphthyl, phenanthryl. ents X and Y as shown below: 35 The term “substituted aryl” refers to aryl as just defined

having one to four Substituents from the group including halogen, CN, OH, NO, amino, alkyl, cycloalkyl, alkenyl, alkynyl, alkoxy, aryloxy, Substituted alkyloxy, alkylcarbonyl, alkylcarboxy, alkylamino, or arylthio. 40 The term “heterocyclic” is used herein to describe a stable 4- to 7-membered monocyclic or a stable multicyclic het erocyclic ring which is Saturated, partially unsaturated, or unsaturated, and which consists of carbon atoms and from X is halogen, CN, methoxy, NO, or 2-thiazole; 45 one to four heteroatoms Selected from the group including Y is a substituent on the 4' or 5" position selected from N, O, and Satoms. The N and Satoms may be oxidized. The H and F; or heterocyclic ring also includes any multicyclic ring in which R" is a five membered ring of the structure: any of above defined heterocyclic rings is fused to an aryl ring. The heterocyclic ring may be attached at any heteroa 50 X tom or carbon atom provided the resultant Structure is chemically Stable. Such heterocyclic groups include, for example, tetrahydrofuran, piperidinyl, piperaZinyl, 2-oxopiperidinyl, azepinyl, pyrrollidinyl, imidazolyl, 55 pyridyl, pyrazinyl, pyrimidinyl, pyridaZinyl, oxazolyl, isoxazolyl, morpholinyl, indolyl, quinolinyl, thienyl, furyl, benzofuranyl, benzothienyl, thiamorpholinyl, thiamorpholi U is O, S, or NH; X is halogen, CN, or NO; nyl Sulfoxide, and isoquinolinyl. The term “substituted heterocyclic” is used herein to Y is H or C to C alkyl; 60 W is O or a chemical bond; describe the heterocyclic just defined having one to four or a pharmaceutically acceptable Salt thereof. Substituents Selected from the group which includes The antiprogestin compounds of this invention may con halogen, CN, OH, NO, amino, alkyl, Substituted alkyl, tain an asymmetric carbon atom and Some of the compounds cycloalkyl, alkenyl, Substituted alkenyl, alkynyl, alkoxy, of this invention may contain one or more asymmetric 65 aryloxy, Substituted alkyloxy, alkylcarbonyl, alkylcarboxy, centers and may thus give rise to optical isomers and alkylamino, or arylthio. The term “alkoxy” is used herein to diastereomers. While shown without respect to stereochem refer to the OR group, where R is alkyl or substituted alkyl. US 6,399,593 B1 15 16 The term “aryloxy” is used herein to refer to the OR group, Solid compositions, particularly tablets and hard-filled or where R is aryl or substituted aryl. The term “alkylcarbonyl liquid-filled capsules. Oral administration of the compounds is used herein to refer to the RCO group, where R is alkyl is preferred. or substituted alkyl. The term “alkylcarboxy” is used herein These active compounds may also be administered to refer to the COOR group, where R is alkyl or substituted parenterally or intraperitoneally. Solutions or Suspensions of alkyl. The term “aminoalkyl” refers to both secondary and these active compounds as a free base or pharmacologically tertiary amines wherein the alkyl or Substituted alkyl groups acceptable Salt can be prepared in water Suitably mixed with containing one to eight carbon atoms, which may be either a Surfactant Such as hydroxypropylcellulose. Dispersions Same or different and the point of attachment is on the can also be prepared in glycerol, liquid, polyethylene glycols nitrogen atom “Halogen” refers to Cl, Br, F, or I. and mixtures thereof in oils. Under ordinary conditions of The antiprogestin compounds of the present invention can Storage and use, these preparations contain a preservative to be used in the form of salts derived from pharmaceutically prevent the growth of microorganisms. or physiologically acceptable acids or bases. These Salts The pharmaceutical forms Suitable for injectable use include, but are not limited to, the following salts with 15 inorganic acids Such as hydrochloric acid, Sulfuric acid, include Sterile aqueous Solutions or dispersions and Sterile nitric acid, phosphoric acid and, as the case may be, Such powders for the extemporaneous preparation of Sterile organic acids as acetic acid, Oxalic acid, Succinic acid, and injectable Solutions or dispersions. In all cases, the form maleic acid. Other Salts include Salts with alkali metals or must be sterile and must be fluid to the extent that easy alkaline earth metals, Such as Sodium, potassium, calcium or Syringe ability exits. It must be stable under conditions of in the form of esters, carbamates and other manufacture and Storage and must be preserved against the conventional “pro-drug forms, which, when administered contaminating action of microorganisms. Such as bacterial in Such form, convert to the active moiety in Vivo. and fungi. The carrier can be a Solvent or dispersion medium These methods may be used for contraception or treat containing, for example, Water, ethanol (e.g., glycerol, pro ment and/or prevention of Secondary amenorrhea, dysfunc 25 pylene glycol and liquid polyethylene glycol), Suitable mix tional bleeding, uterine leiomyomata, endometriosis, poly tures thereof, and vegetable oil. cystic ovary Syndrome, carcinomas and adenocarcinomas of The antiprogestin compounds of this invention can be the endometrium, Ovary, breast, colon, prostate, or minimi prepared following the Schemes illustrated below: Zation of Side effects or cyclic menstrual bleeding. Addi AS demonstrated in Scheme I, the antiprogestin com tional uses of the invention include stimulation of food pounds of this invention are generally prepared by employ intake. ing the Suitable coupling reaction as a final Step. An appro When the compounds are employed for the above priately Substituted ortho-amino acid or its derivatives Such utilities, they may be combined with one or more pharma as ethyl ester (X= Br, I, Cl, or a latent coupling precursor ceutically acceptable carriers or excipients, for example, 35 Such as alkoxy group which can be converted into OTf group Solvents, diluents and the like, and may be administered Suitable in the coupling reaction) is treated with a Suitable orally in Such forms as tablets, capsules, dispersible organo metallic reagent, e.g. Grignard reagent, in appropri powders, granules, or Suspensions containing, for example, ate nonprotic Solvents which include but are not limited to from about 0.05 to 5% of suspending agent, syrups THF or ether to give ortho-amino carbinol 2 under an inert containing, for example, from about 10 to 50% of Sugar, and 40 atmosphere Such as argon or nitrogen at -78 C. to room elixirs containing, for example, from about 20 to 50% temperature. Ring closure of carbinol 2 to yield oxazin-2- ethanol, and the like, or parenterally in the form of Sterile ones 3 is commonly effected by a condensing agent Such as injectable Solutions or Suspensions containing from about carbonyldiimidazole, phosgene, dimethylcarbonate, or 0.05 to 5% Suspending agent in an isotonic medium. Such 45 diethylcarbonate in a suitable nonprotic solvent such as THF pharmaceutical preparations may contain, for example, from at temperatures ranging from room temperature to 65 C. about 25 to about 90% of the active ingredient in combina The arylation of oxazin-2-ones 3 to yield 4 can be effected tion with the carrier, more usually between about 5% and by various coupling reactions including Suzuki, Stille reac 60% by weight. tions etc. These reactions are commonly performed in the These active agents of the methods and regimens herein 50 presence of transition metallic catalyst, e.g., palladium or may be administered orally as well as by intravenous, nickel complex often with phosphino ligands, e.g., PhP, intramuscular, or Subcutaneous routes. Solid carriers include 1, 1'-bis(diphenylphosphino) ferrocene, 1,2-bis Starch, lactose, dicalcium phosphate, microcrystalline (diphenylphosphino)ethane or palladium salts such as pal cellulose, Sucrose and kaolin, while liquid carriers include 55 ladium acetate. Under this catalytic condition, an appropri Sterile water, polyethylene glycols, non-ionic Surfactants and ately Substituted nucleophilic reagent, e.g., aryl boronic edible oils. Such as corn, peanut and Sesame oils, as are acid, arylstannane, or aryl compound, is coupled with appropriate to the nature of the active ingredient and the oxazinones 3 to give compounds 4. If a base is needed in the particular form of administration desired. Adjuvents cus reaction, the commonly used bases include but not limited to tomarily employed in the preparation of pharmaceutical 60 Sodium bicarbonate, Sodium carbonate, potassium compositions may be advantageously included, Such as phosphate, barium carbonate, potassium acetate, or cesium flavoring agents, coloring agents, preserving agents, and fluoride. The most commonly used Solvents in these reac antioxidants, for example, Vitamin E, ascorbic acid, BHT tions include benzene, DMF, isopropanol, ethanol, DME, and BHA. 65 ether, acetone or a mixture of any one of these Solvent and The preferred pharmaceutical compositions from the water. The coupling reaction is generally executed under an Standpoint of ease of preparation and administration are inert atmosphere Such as nitrogen or argon at temperatures US 6,399,593 B1 17 18 ranging from room temperature to 95 C. Oxazinones 3 can non-protic Solvent Such as toluene, chlorobenzene, benzene, be converted into a nucleophile Such as boronic acid which or Xylene under an inert atmosphere Such as argon or can be coupled with an appropriate electrophile, e.g., aryl nitrogen at the temperature of boiling Solvent. bromide or aryl iodide, to yield 6 employing the coupling reaction condition as described above. Scheme II describes the procedures to prepare oxazinones bearing two different substituents at position-4. The Weinreb Scheme I amide 9 can be prepared from an appropriately Substituted is a to ic anhydride when treated with N-, O X A. COOR dimethylhydroxyl-amine hydrochloride Salt in a protic Sol 21 RMgBr, THF, rt, N2 vent Such as ethanol or isopropanol at reflux under an inert Bn. H atmosphere Such as argon or nitrogen. Coupling of amide 9 n NH2 with an aryl electrophile Such as aryl boronic acid or 1. arylstannane to give 10 can be effected by employing a R R 15 typical coupling reaction Such as Suzuki, Stille coupling X A. 21 OH procedure in a similar fashion as described for the prepara CDI, THF, 50 degrees C., N2 tion of oxazinones 4. Treatment of Weinreb amide 10 with He Bn organo metallic compounds, e.g., alkyllithium, s NH2 alkynyllithium, aryllithium, or their Grignard counterpart in 2 a nonprotic solvent such as TBF or ether under an inert R R atmosphere Such as argon or nitrogen at -78 C. to room X A. 21 O ArB(OH)2, Pd(Ph3P)4, Na2COs temperature affords amino ketone 11. Conversion of ketone 1s DME/H2O, N, 85 degrees C. 11 to carbinol 12 can be effected by treatment of 10 with an BS 25 s N O organo metallic reagent Such as alkyl, alkynyl, or aryl H Grignard compound in a nonprotic Solvent Such as THE or 3 ether under an inert atmosphere Such as argon or nitrogen at R R -78 C. to room temperature. Conversion of ketone 11 to Air A. carbinol 12 can also be effected by reduction of the ketone 21 O n-BuLi, THF, B(OMe)3 group of 11 to the carbinol moiety of 12 using an appropriate Bn 1s -78 degrees C. rt, N2 se s N O reducing reagent Such as lithium aluminum hydride, Sodium H borohydride in a suitable solvent such as THF, ether, or 4 anhydrous alcohol under an inert atmosphere in the tem R R 35 perature ranging from O C. to the boiling point of the B(OH)2 A. Solvent. Ring closure of carbinol 12 to produce the com Y O ArBr, Pd(Ph3P)4, Na2CO3 pounds of this invention, 13, can be accomplished with BS -N HossDME/H2O, 85 degrees C., N2 condensing agents Such as carbonyldiimidazole, phosgene, s N O H 40 dimethylcarbonate, or diethylcarbonate in a Suitable non 5 protic Solvent Such as THF at temperatures ranging from R R room temperature to 65 C. Conversion of carbamate 13 to Air A. thiocarbamate 14 can be readily effected by treatment of 13 21 O Lawesson's reagent with a Suitable Sulfur reagent Such as PSs or Lawesson's He 45 BS reagent in a Suitable nonprotic Solvent Such as toluene, n N O chlorobenzene, benzene, or Xylene under an inert atmo H Sphere Such as argon or nitrogen at the temperature of R R boiling Solvent. 50 Air A. NY O Scheme II O Bns Nls S H X A. 7 55 21 O 1s EtOH, NHOMeMe, HCl, reflux BS The transformation of 3 to 5 can be effected by treating 3 n N O with an organo metallic reagent, e.g., n-BuLi, in a nonprotic H solvent such as THF or ether followed by quenching the 8 reaction Solution wiht a Suitable electrophile Such as trime 60 O thil borate, triisopropyl borate, bisheXalkyl tin reagent, or X A. /O 21 NN Zinc chloride at temperatures ranging from -78 C. to room ArB(OH)2, Pd(Ph3P)4, Na2COs temperature under an inert atmosphere Such as argon or BN DME/H2O, 85 degrees C., N2 nitrogen. Conversion of carbamate 6 to thiocarbamate 7 can 65 D NH2 be readily effeted by treatment of 6 with a suitable sulfur 9 reagent Such as PSs or Lawesson's reagent in a Suitable US 6,399,593 B1 20 -continued -continued O Air A. /O Y NN R.Li or RMgX, THF, -78 degrees C. to rt BN D NH2 1O O 1O The following non-limiting examples illustrate the com Air A. 21 R6 pounds useful in the invention. R7MgX, -78 degrees C. to rt, N2 Her EXAMPLE 1. Ban or reducing reagent D NH2 2-Amino-5-bromo-3-pyridine Carboxylic Acid 11 15 To a solution of 2-amino-nicotinic acid (10g, 72.5 mmol) OH R7 in acetic acid (70 mL) was dropwise added bromine (9.8 mL, Air A. 79.8 mmol) at room temperature under nitrogen. Upon 21 R6 CDI or triphosgene, THF completion of the reaction, the Solvent was removed in 0 degrees C. to 65 degrees C. Bn H vacuo, the residue triturated with ether and collected on a s NH2 filter to give 2-amino-5-bromo-3-pyridine carboxylic acid as 12 a yellow solid (15.7g, 99%); mp 272 C., (decomposed); R6 R7 H-NMR (DMSO-d) & 8.8–7.8 (bs, 2H), 8.44 (d. 1H, Air A. J=2.48 Hz), 8.34 (d. 1H, J=2.48 Hz); MS (EI) m/z 216/218 21 O 25 (IM+H, 100%). 1. HerLawesson's reagent EXAMPLE 2 BS n N O H 2-(2-Amino-5-bromo-pyridin-3-yl)-propan-2-ol 13 To a solution of 2-amino-5-bromo-3-pyridine carboxylic R6 R7 acid (5 g, 23 mmol) in THF (70 mL) at 0° C. was added methyl magnesium bromide (13.7 g, 115 mmol) under Air A. nitrogen. After addition, the reaction mixture was heated at Y O 65 C. for 12 hours, cooled to room temperature and quenched with Saturated aqueous ammonium chloride. The BSs N 1. S 35 H ethyl acetate was added and organic layer was separated, 14 dried over Sodium Sulfate and concentrated. The residue was purified via flash chromatography to give 2-(2-amino-5- bromo-pyridin-3-yl)propan-2-ol as a yellow Solid (1.2 g, 23%); mp 107–108° C.; H-NMR (DMSO-d) & 7.89 (d. 1H, Alternatively, ortho-amino ketone 11 can be prepared by 40 J=2.3 Hz), 7.40 (d. 1H, J=2.3 Hz), 6.28 (bs, 2H), 5.51 (s, treatment of ortho-amino nitrile 16 with an organo metallic 1H), 1.4 (s, 6H); MS (APCI) m/z 231 (IM+H", 100%). compound Such as organo lithium reagent or Gringard EXAMPLE 3 reagent in a suitable solvent such as THF or ether under an 6-Bromo-4,4'-dimethyl-1,4-dihydro-3-oxa-1,8-diaza inert atmosphere Such as argon or nitrogen at temperatures 45 naphthlalen-2-one ranging from -78 C. to room temperature as illustrated in A mixture of 2-(2-amino-5-bromo-pyridin-3-yl)-propan Scheme III. Nitrile 16 can be readily prepared from an 2-ol (0.86 g., 3.7 mmol) and 1,1'-carbonyldiimidazole appropriately Substituted nitrite Such as bromobenzonitrile (excess) in THF (10 mL) was heated at 35° C. overnight. The 15 using a Suitable coupling reaction Such as Stille or Suzuki 50 reaction Solution was cooled to room temperature, poured protocol carried out in a Similar fashion as described for the into ethyl acetate (200 mL), washed with 1 NHCl (2x50 mL), dried over Sodium Sulfate, and concentrated. The preparation of the Weinreb amide 10. residue was purified by a flash chromatography (silica gel, 25% ethyl acetate/hexane) to afford 6-bromo-4,4'-dimethyl Scheme III 55 1,4-dihydro-3-oxa-1,8-diaza-naphthalen-2-one (0.9 g, 94%) X A. CN as a white solid: mp 251-252 C.; H-NMR (DMSO-d) & ArB(OH)2. Na2COs 10.9 (s, 1H), 8.32 (d. 1H, J=2.19 Hz), 8.0 (d. 1H, J=2.22 Hz), Her YBn C Pd(0), DME/HO, N. 1.6 (s, 6H); MS (EI) m/z 256 (M", 80%); Anal. Calc. For s NH2 60 CHBrNO: C, 42.05, H, 3.53, N, 10.90. Found: C, 42.15, 15 H, 3.65, N, 10.80. Air A. CN EXAMPLE 4 R6 Lior R6MgX, 0 degrees C. 6-(3-Chloro-phenyl)-4,4-dimethyl-1,4-dihydro-3- NYBn. C n NH2 65 oxa-1,8-diaza-naphthalene-2-one 16 A mixture of 6-bromo-4,4'-dimethyl-1,4-dihydro-3-oxa 1,8-diaza-naphthalen-2-one (0.1 g, 0.39 mmol), US 6,399,593 B1 21 22 3-chlorophenylboronic acid (0.075 g, 0.47 mmol), tetrakis nitrogen was added a Solution of methylmagnesium bromide (triphenylphosphine)-palladium (0) (0.023 g, 0.02 mmol), (3M in ether, 8.1 mL, 24.3 mmol). The reaction mixture was and sodium carbonate (0.1 g, 0.94 mmol) in DME (8 mL) stirred for 30 minutes and then slowly quenched with H2O, and water (5 mL) was subject to a blanket of nitrogen for 15 treated with 5N HCl Solution. The mixture was extracted minutes at 50° C. and then was heated at 85° C. for 30 with ethyl acetate (3x100 mL) and organic extracts were minutes. The reaction was cooled to room temperature and ethyl acetate (100 mL) was added. The organic layer washed washed with brine, and dried (MgSO). After removal of with aqueous ammonium chloride (2x50 mL) and with brine Solvent, the residue was purified by chromatography using (50 mL), dried over magnesium Sulfate and concentrated. EtOAc/hexane mixture (1:3) as eluent to afford the title The product was dissolved in dichloromethane and passed compound as a greenish crystalline Solid: (0.71g, 85%): mp: through a plug of magneSol. The Solvent was removed and 108-110° C. "H NMR (DMSO-d) & 2.51 (s.3H), 7.28–7.39 the clear oil obtained triturated with ether (25 mL) to give (m, 4H); MS(ESI) m/z. 171/173 (M+H)"; Anal. Calc. For 6-(3-chloro-phenyl)-4,4-dimethyl-1,4-dihydro-3-oxa-1,8- C.H.CINO: C, 49.28, H, 4.14, N, 16.14. Found: C, 49.70, diaza-naphthalene-2-one as a white solid (0.087g,80%): mp 15 H, 4.03, N, 16.41. 214–215° C.; H-NMR (DMSO-d) & 10.9 (s, 1H), 8.56 (d. 1H, J-2.35 Hz), 8.06 (d. 1H, J=2.35 Hz), 7.86 (t, 1H, J=2.35 EXAMPLE 8 Hz), 7.72 (td, 1H, J=9.4, 2.35 Hz), 7.54-7.44 (m, 2H), 1.69 (s, 6H); MS (ESI) m/z 289 (IM+H, 100%); Anal. Calc. For 1-(3-Amino-6-chloro-pyridin-2-yl)-propan-2-ol C.H.NO: C, 62.40, H, 4.54, N, 9.70. Found: C, 60.53, H, 4.40, N, 9.21. To a solution of 1-(3-amino-6-chloro-pyridin-2-yl)- EXAMPLE 5 ethanone in anhydrous THF under N2 was added a solution 25 of methylmagnesium bromide (3N in ether). The resulting 6-Chloro-3-nitro-pyridine-2-carbonitrile reaction mixture was stirred at room temperature for 4 h, A mixture of 2,6-dichloro-3-nitropyridine and cuprous then slowly quenched with HO, treated with 0.5 NHCl and cyanide in 1-methyl-2-pyrrolidinone was quickly heated to stirred for 30 minutes, diluted with ethyl acetate, washed 180° C. and maintained at that temperature for 15 minutes with brine, dried (MgSO4), concentrated, and chromato with vigorous stirring. The mixture was cooled to -10 C. graphed using a mixture of EtOAc/Hexane (3.5: 6.5) to and the deep brown Solution was poured into ice water (3.5 afford the title compound as a white crystals (0.45 g, 82%); L) and stirred for 30 min. The flocculent brown precipitate mp: 118–121° C. "H NMR(DMSO-d) & 1.45(s, 6H), 3.35(s, was collected and washed with H2O. After drying for about 1H), 5.51(s, 1H), 5.68(s, 1H), 7.02(s, 1H); MS((+)APCI) 1.5 h, the moist Solid was extracted with boiling toluene 35 m/z 187/189 (M+H)"; Anal. Calc. For CHCINO: C, (3x300 mL). The combined toluene extracts were washed 51.48, H, 5.94, N, 15.01. Found: C, 51.22, H, 5.99, N, 14.47. with H2O, brine, and dried (MgSO), concentrated. The crude product was crystallized from EtOAc/hexane to afford EXAMPLE 9 the title compound: mp 115-117 C.; H NMR(DMSO-d) 40 & 8.16 (dd, 1H, J=8.7, 3.0 Hz), 8.82 (d. 1H, J=9.0 Hz); MS 6-Chloro-4,4-dimethyl-1,4-dihydro-3-oxa-1,5-diaza (EI) m/z 183/185 (M+H)"; Anal. Calc. For CHCINO: C, naphthalen-2-one 39.26, H, 1.10, N, 22.89. Found: C, 39.47, H, 1.38, N, 22.77. To a solution of 1-(3-amino-6-chloro-3-nitro-pyridin-2- EXAMPLE 6 45 yl)-propan-2-ol (0.3g, 1.67 mmol) in anhydrous THF (20 3-Amino-6-chloro-pyridine-2-carbonitrile mL) was added a Solution of 1,1'-carbonyldiimidazole To a solution of 2,6-dichloro-3-nitropyridine (4.8 g., 26.15 (0.68g, 4.12 mmol) in anhydrous THF (10 mL). The reaction mmol) in MeOH (60 mL) and concentrated HCl (25 mL) mixture was heated under reflux for 3 h. The reaction was slowly added iron powder (5.12 g, 91.53 mmol). After 50 mixture was treated with 0.5N HCl, washed with brine, the completion of addition, the mixture was refluxed for 45 HO, dried (MgSO), concentrated and crystallized from minutes and poured into 700 mL of H.O. Filtration of the EtOAc/hexane to obtain the title compound as a white resulting Slurry gave a dull yellow Solid. The filtrate was crystalline solid (0.2g, 56%): mp 175-178° C. "H NMR made basic with concentrated NHOH, the resulting slurry 55 (DMSO-d) & 1.60 (s, 6H), 7.30 (d. 1H, J=8.41 Hz), 7.41 (d. was filtered and both the Solid and the filtrates were 1H, J=8.41 Hz), 10.47 (s, 1H); MS((+)APCI) m/z 213 extracted with ether. The combined extracts were dried (M+H)"; Anal. Calc. For CHCINO: C, 50.84, H, 4.27, N, (MgSO) and evaporated to give the title compound as a 13.17. Found: C, 50.99, H, 4.28, N, 12.98. creamy solid (2.8 g., 58%); mp 162-165 C. which was used in next step without further purification. 60 EXAMPLE 10 EXAMPLE 7 The compounds of this invention were tested in the 1-(3-Amino-6-chloro-pyridin-2-yl)-ethanone relevant assay as described below and their potency are in 65 the range of 0.01 nM to 5 uM in the in vitro assays and 0.001 To a solution of 3-amino-6-chloro-pyridine-2-carbonitrile to 300 mg/kg in the in Vivo assays. The Selected example is (0.75g, 4.88 mmol) in anhydrous THF (25 mL) under example 4. US 6,399,593 B1 23 24 progesterone. Twenty-four hr. after treatment, the medium is C discarded, cells are washed three times with D-PBS (GIBCO, BRL). Fifty ul of cell lysis buffer (Promega, Madison, Wis.) is added to each well and the plates are Me Me shaken for 15 min in a Titer Plate Shaker (Lab Line Instrument, Inc.). Luciferase activity is measured using luciferase reagents from Promega. rry c. Analysis of Results N 2 N 1sO Each treatment consists of at least 4 replicates. Log H transformed data are used for analysis of variance and nonlinear dose response curve fitting for both agonist and Example 4, hPR CV-1, IC=0.8 uM antagonist modes. Huber weighting is used to downweight A. In-vitro Biology The in-vitro biology is determined by (1) competitive the effects of outliers. ECso or ICso values are calculated Radioligand Binding: using the A-form of the human 15 from the retransformed values. JMP software (SAS Institute, progesterone receptor with progesterone as the radioligand; Inc.) is used for both one-way analysis of variance and (2) co-transfection assay, which provides functional activity nonlinear response analyses. expressed as agonist EC50 and Antagonist IC50 values; (3) d. Reference Compounds a T47D cell proliferation, which is a further functional assay Progesterone and trimegestone are reference progestins which also provides agonist and antagonist data; and (4) and RU486 is the reference antiprogestin. All reference T47D cell alkaline phosphatase assay, which is a further compounds are run in full dose-response curves and the functional assay which also provides agonist and antagonist ECso or ICso values are calculated. data. 1. hPR Binding Assay 25 TABLE 1. This assay is carried out in accordance with: Pathirana, Estimated ECso, standard error (SE), and 95% confidence intervals C.; Stein, R. B.; Berger, T. S.; Fenical, W.; Ianiro, T., Mais, CI) for reference progestins from three individual studies D. E.; Torres, A, Glodman, M. E., NonSteroidal human EC50 95% CI progesterOne receptor modulators from the marine alga cymoplia barbata, J. Steroid Biochem Mol. Biol., 1992, 41, Compound Exp. (nM) SE lower upper 733 738. Progesterone 1. O. 616 O.O26 O.509 O.746 2 O.4O2 O.O19 O.323 OSO1 2. PRE-luciferase ASSay in CV-1 Cells 3 O.486 O.O28 O.371 O.637 The object of this assay is to determine a compounds Trimegestone 1. O.OO75 O.OOO2 O.OO66 O.OO85 progestational or antiprogestational potency based on its 35 2 O.OO81 O.OOO3 O.OOFO O.OO94 effect on PRE-luciferase reporter activity in CV-1 cells 3 O.OO67 O.OOO3 O.OO55 O.OO82 co-transfected with human PR and PRE-luciferase plasmids. The materials methods used in the assay are as follows. a. Medium TABLE 2 40 The growth medium was as follows: DMEM (Bio Estimated ICso standard error (SE), and 95% confident interval (CI) Whittaker) containing 10% (v/v) fetal bovine serum (heat for the antiprogestin, RU486 from three individual studies inactivated), 0.1 mM MEM non-essential amino acids, 100 U/ml penicillin, 100 mg/ml streptomycin, and 2 mM IC 50 95% CI GlutaMax (GIBCO, BRL). The experimental medium was 45 Compound Exp. (nM) SE lower upper as follows: DMEM (Bio Whittaker), phenol red-free, con RU486 1. O.O28 O.OO2 O.O19 O.O42 taining 10% (v/v) charcoal-stripped fetal bovine serum 2 O.O37 O.OO2 O.O29 O.O48 (heat-inactivated), 0.1 mM MEM non-essential amino acids, 3 O.O19 O.OO1 O.O13 0.027 100 U/ml penicillin, 100 mg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL). 50 Progestational activity: Compounds that increase PRE b. Cell Culture, Transfection, Treatment, and Luciferase luciferase activity significantly (p<0.05) compared to ASSay vehicle control are considered active. Stock CV-1 cells are maintained in growth medium Antiprogestational activity: Compounds that decrease 3 Co-transfection is done using 1.2x107 cells, 5 mg plEM 55 nM progesterone induced PRE-luciferase activity signifi plasmid with hPR-B inserted at Sph1 and BamH1 sites, 10 cantly (p<0.05) mg pGL3 plasmid with two PREs upstream of the luciferase ECso: Concentration of a compound that gives half Sequence, and 50 mg Sonicated calf thymus DNA as carrier maximal increase PRE-luciferase activity (default-nM) with DNA in 250 ml. Electroporation is carried out at 260 V and SE 1,000 m F in a Biorad Gene Pulser II. After electroporation, 60 ICso: Concentration of a compound that gives half cells are resuspended in growth medium and plated in maximal decrease in 3 nM progesterone induced PRE 96-well plate at 40,000 cells/well in 200 ul. Following luciferase activity (default-nM) with SE. overnight incubation, the medium is changed to experimen 3. T47D Cell Proliferation Assay tal medium. Cells are then treated with reference or test 65 The objective of this assay is the determination of proges compounds in experimental medium. Compounds are tested tational and antiprogestational potency by using a cell for antiprogestational activity in the presence of 3 nM proliferation assay in T47D cells. A compound's effect on US 6,399,593 B1 25 26 DNA synthesis in T47D cells is measured. The materials and methods used in this assay are as follows. TABLE 3 a. Growth Medium Estimated ECso, standard error (SE), and 95% confidence intervals CI) for individual studies DMEM:F12 (1:1) (GIBCO, BRL) supplemented with 5 10% (v/v) fetal bovine serum (not heat-inactivated), 100 U/ml penicillin, 100 mg/ml streptomycin, and 2 mM ECso 95% CI GlutaMax (GIBCO, BRL). Compound Exp (nM) SE lower upper b. Treatment Medium Trimegestone 1. O.O17 O.OO3 O.OO7 O.O40 2 O.O14 O.OO1 O.O11 O.O17 Minimum Essential Medium (MEM) (#51200 3 O.O19 O.OO1 O.O16 O.O24 038GIBCO, BRL) phenol red-free supplemented with 0.5% MPA 1. O.O19 O.OO1 O.O13 0.027 charcoal stripped fetal bovine serum, 100 U/ml penicillin, 2 O.O17 O.OO1 O.O11 O.O24 200 mg/ml streptomycin, and 2 mM GlutaMax (GIBCO, BRL). 15 c. Cell Culture TABLE 4 Stock T47D cells are maintained in growth medium. For Estimated ICs, standard error, and 95% confident interval for the BrdU incorporation assay, cells are plated in 96-well plates antiprogestin, RU486 (Falcon, Becton Dickinson Labware) at 10,000 cells/well in ICso 95% CI growth medium. After overnight incubation, the medium is changed to treatment medium and cells are cultured for an Compound Exp (nM) SE lower upper additional 24 hr before treatment. Stock compounds are RU486 1. O.O11 O.OO1 O.OO8 O.O14 2 O.O16 O.OO O.O14 O.O2O dissolved in appropriate vehicle (100% ethanol or 50% 3 O.O18 O.OOI O.O14 O.O22 ethanol/50% DMSO), Subsequently diluted in treatment 25 medium and added to the cells. Progestin and antiprogestin reference compounds are run in full dose-response curves. ECso: Concentration of a compound that gives half-maximal increase in BrdU incorporation with SE; ICso: Concentration The final concentration of vehicle is 0.1%. In control wells, of a compound that gives half-maximal decrease in 0.1 cells receive vehicle only. Antiprogestins are tested in the trimegestone induced BrdU incorporation with SE presence of 0.03 nM trimegestone, the reference progestin 4. T47D Cell Alkaline Phosphatase Assay agonist. Twenty-four hours after treatment, the medium is The purpose of this assay is to identify progestins or discarded and cells are labeled with 10 mM BrdU antiprogestins by determining a compound's effect on alka (Amersham Life Science, Arlington Heights, Ill.) in treat line phosphatase activity in T47D cells. The materials and ment medium for 4 hr. 35 methods used in this assay are as follows. d. Cell Proliferation Assay a. Culture Medium DMEM:F12 (1:1) (GIBCO, BRL) supplemented with 5% At the end of BrdU labeling, the medium is removed and (v/v) charcoal Stripped fetal bovine Serum (not heat BrdU incorporation is measured using a cell proliferation inactivated), 100 U/ml penicillin, 100 tug/ml streptomycin, ELISA kit (#RPN250, Amersham Life Science) according 40 and 2 mM GlutaMax (GIBCO, BRL). to manufacturers instructions. Briefly, cells are fixed in an b. Alkaline Phosphatase Assay Buffer ethanol containing fixative for 30 min, followed by incuba I. 0.1 M Tris-HCl, pH 9.8, containing 0.2% Triton tion in a blocking buffer for 30 min to reduce background. X-100II. 0.1 M Tris-HCl, pH 9.8 containing 4 mM Peroxidase-labeled anti-BrdU antibody is added to the wells p-nitrophenyl phosphate (Sigma). and incubated for 60 min. The cells are rinsed three times 45 c. Cell Culture and Treatment with PBS and incubated with 3,3'5,5'-tetramethylbenzidine Frozen T47D cells were thawed in a 37° C. water bath and (TMB) Substrate for 10–20 mindepending upon the potency diluted to 280,000 cells/ml in culture medium. To each well of tested compounds. Then 25 ul of 1 M Sulfuric acid is in a 96-well plate (Falcon, Becton Dickinson Labware), 180 added to each well to Stop color reaction and optical density 50 All of diluted cell Suspension was added. Twenty ul of is read in a plate reader at 450 nm within 5 min. reference or test compounds diluted in the culture medium e. Analysis of Results was then added to each well. When testing for progestin antagonist activity, reference antiprogestins or test com Square root-transformed data are used for analysis of pounds were added in the presence of 1 nM progesterone. variance and nonlinear dose response curve fitting for both 55 The cells were incubated at 37° C. in a 5% CO/humidified agonist and antagonist modes. Huber weighting is used to atmosphere for 24 hr. downweight the effects of outliers. ECso or ICso values are d. Alkaline Phosphatase Enzyme ASSay calculated from the retransformed values. JMP Software At the end of treatment, the medium was removed from (SAS Institute, Inc.) is used for both one-way analysis of the plate and fifty ul of assay buffer I was added to each well. variance and non-linear dose response analyses in both 60 The plates were shaken in a titer plate Shaker for 15 min. Single dose and dose response Studies. Then 150 ul of assay buffer II was added to each well. f. Reference Compounds Optical density measurements were taken at 5 min intervals Trimegestone and acetate (MPA) for 30 min at a test wavelength of 405 nM. are reference progestins and RU486 is the reference anti 65 e. Analysis of Results: Analysis of DoSe-response Data progestin. All reference compounds are run in full dose For reference and test compounds, a dose response curve response curves and the ECso or ICso values are calculated. is generated for dose (X-axis) vs. the rate of enzyme reaction US 6,399,593 B1 27 28 (slope) (Y-axis). Square root-transformed data are used for Farms, N.Y.) following surgery. Ovariectomy is performed analysis of variance and nonlinear dose response curve at least 10 days prior to treatment to reduce circulating SeX fitting for both agonist and antagonist modes. Huber weight . Animals are housed under 12 hr light/dark cycle ing is used to downweight the effects of outliers. ECso or and given Standard rat chow and water ad libitum. ICso values are calculated from the retransformed values. JMP software (SAS Institute, Inc.) is used for both one-way c. Treatment analysis of variance and non-linear dose response analyses Rats are weighed and randomly assigned to groups of 4 or in both Single dose and dose response Studies. 5 before treatment. Test compounds in 0.2 ml vehicle are Reference Compounds: administered by Subcutaneous injection in the nape of the Progesterone and trimegestone are reference progestins neck or by gavage using 0.5 ml. The animals are treated once and RU486 is the reference antiprogestin. All reference daily for Seven days. For testing antiprogestins, animals are compounds are run in full dose response curves and the ECso given the test compounds and a ECso dose of progesterone or ICso values are calculated. (5.6 mg/kg) during the first three days of treatment. Follow 15 ing decidual Stimulation, animals continue to receive TABLE 5 progesterone until necropsy four days later. Estimated ECso standard error (SE), and 95% confidence intervals d. Dosing CI) for reference progestins from three independent experiments Doses are prepared based upon mg/kg mean group body EC50 95% CI weight. In all Studies, a control group receiving vehicle is Compound Exp. (nM) SE lower upper included. Determination of dose-response curves is carried out using doses with half log increases (e.g. 0.1, 0.3, 1.0, 3.0 Progesterone 1. O.839 O.O3O O.7O6 O.996 2 O.639 O.OO6 O. 611 O.669 mg/kg. . . ). 3 1.286 0.029 1.158 1.429 25 e. Decidual Induction Trimegestone 1. O.O84 O.OO2 O.O76 O.O91 2 O.O76 O.OO1 O.O72 O.O8O Approximately 24 hr after the third injection, decidual 3 O.16O O.OO4 O.141. O.181 ization is induced in one of the uterine horns by Scratching the antimesometrial luminal epithelium with a blunt 21 G needle. The contralateral horn is not Scratched and Serves as TABLE 6 an unstimulated control. Approximately 24 hr following the Estimated ICs, standard error, and 95% confident interval for the final treatment, rats are sacrificed by CO asphyxiation and reference antiprogestin RU486 from three independent expeXiments body weight measured. Uteri are removed and trimmed of fat. Decidualized (D-horn) and control (C-horn) uterine IC50 95% CI 35 horns are weighed Separately. Compound Exp (nM) SE lower upper f. Analysis of Results RU486 1. O.103 O.OO2 O.O92 O.115 The increase in weight of the decidualized uterine horn is 2 O.120 O.OO1 O. 115 O.126 3 O.O94 O.OO7 O.O66 O.134 calculated by D-horn/C-horn and logarithmic transformation 40 is used to maximize normality and homogeneity of variance. The Huber M-estimator is used to down weight the outlying B. In-vivo Biology transformed observations for both dose-response curve fit The primary in-vivo assay is the rat decidualization ting and one-way analysis of variance. JMP software (SAS model, which may be used to determine progestational Institute, Inc.) is used for both one-way ANOVA and non effects of both agonists and antagonists. The Secondary 45 linear dose-response analyses. in-vivo assay is the rat ovulation inhibition model, which is under development, and hence the protocol is un-available. g. Reference Compounds 1. Rat Decidualization ASSay All progestin reference compounds were run in full dose The objective of this procedure is used to evaluate the response curves and the ECso for uterine wet weight were effect of progestins and antiprogestins on rat uterine decidu 50 calculated. alization and compare the relative potencies of various test compounds. The materials and methods used in this assay TABLE 7 are as follows. Estimated ECs standard error (SE), and 95% confidence intervals for a. Methods 55 individual studies Test compounds are dissolved in 100% ethanol and mixed with corn oil (vehicle). Stock solutions of the test com ECso 95% CI pounds in oil (Mazola"M) are then prepared by heating (-80 Compound Exp (mg/kg S.C.) SE lower upper C. the mixture to evaporate ethanol. Test compounds are Progesterone 1. 5.50 O.77 4.21 7.20 Subsequently diluted with 100% corn oil or 10% ethanol in 60 2 6.21 1.12 4.41 8.76 3-Ketodesogestrel 1. O.11 O.O2 O.O7 O16 corn oil prior to the treatment of animals. No difference in 2 O.10 O.OS O.11 O.25 decidual response was found when these two vehicles were 3 O.O6 O.O3 O.O3 O.14 compared. Levonorgestrel 1. O.O8 O.O3 O.04 O16 2 O.12 O.O2 O.09 0.17 b. Animals (RACUC Protocol #5002) 65 3 O.O9 O.O2 O.O6 O.13 Ovariectomized mature female Sprague-Dawley rats 4 O.O9 O.O2 O.O6 O.14 (~60-day old and 230 g) are obtained from Taconic (Taconic US 6,399,593 B1 29 30 What is claimed: TABLE 7-continued 1. A method of contraception, which comprises adminis Estimated ECso standard error (SE), and 95% confidence intervals for tering to a female of child bearing age for 28 consecutive individual studies days: 5 ECso 95% CI a) a first phase of from 14 to 24 daily dosage units of a progestational agent equal in progestational activity to Compound Exp (mg/kg S.C.) SE lower upper about 35 to about 100 lug levonorgestrel; MPA 1. O.42 O.O3 O.29 O.60 b) a Second phase of from 1 to 11 daily dosage units, at 2 O.39 O.OS O.22 O.67 1O a daily dosage of from about 2 to 50 mg, of an 3 O.39 O.04 O.25 O.61 antiprogestin compound of Formula I:

TABLE 8 15 R1 R2 Estimated average EC50, standard error, and 95% confidence R A. intervals for dose-response curves of 3 reference compounds n W EC50 95% CI B 2 1s Compound (mg/kg, s.c.) SE lower upper N N O Progesterone 5.62 O.62 4.55 7.00 k 3-Ketodesogestrel O.10 O.O2 O.O7 O.14 Levonorgestrel O.10 O.O1 O.O8 O.12 wherein: 25 A, B and D are N or CH, with the proviso that A, B and D can not all be CH; TABLE 9 R" and R are independent substituents selected from Estimated ICs, standard error, and 95% confident interval for the the group consisting of H, C to C alkyl, Substituted antiprogestin, RU 486 C to C alkyl, C. to Calkenyl, Substituted C to C ICso 95% CI alkenyl, C. to C alkynyl, Substituted C to C alkynyl, C. to Cs cycloalkyl, Substituted C to Cs Compound Exp. (mg/kg p.o.) SE lower upper cycloalkyl, aryl, Substituted aryl, heterocyclic, Sub RU486 1. O.21 O.O7 O.OS O.96 stituted heterocyclic, COR, and NRCOR; 2 O.14 O.O2 O.O8 0.27 or R' and R are fused to form a spirocyclic ring 35 Selected from the group consisting of (i), (ii), and (iii): Concentration: Compound concentration in assay (i) a carbon-based Saturated 3 to 8 membered Spiro (default-mg/kg body weight) cyclic ring; Route of administration: Route the compound is admin 40 (ii) a carbon-based 3 to 8 membered Spirocyclic ring istered to the animals having one or more carbon-carbon double bonds, Body weight: Mean total animal body weight (default-kg) and D-horn: Wet weight of decidualized uterine horn (default (iii) a 3 to 8 membered heterocyclic Spirocyclic ring mg) containing in its backbone one to three heteroat 45 oms Selected from the group consisting of O, S C-horn: Wet weight of control uterine horn (default-mg) and N, Decidual response: (D-C)/Cx100% the Spirocyclic rings of (i), (ii), and (iii) being option Progestational activity: Compounds that induce decidu ally substituted by from 1 to 4 groups selected from alization significantly (p<0.05) compared to vehicle the group consisting of fluorine, C to C alkyl, C to control are considered active 50 C alkoxy, C to C thioalkyl, CF, OH, CN, NH, Antiprogestational activity: Compounds that decrease NH(C. to C alkyl), and N(C. to C alkyl); ECso progesterone induced decidualization signifi R is H, C, to C alkyl, substituted C to C alkyl, aryl, cantly (p<0.05) Substituted aryl, C to Calkoxy, Substituted C to C. ECso for uterine weight: Concentration of compound that alkoxy, C to Caminoalkyl, or Substituted C to C. gives half-maximal increase in decidual response 55 aminoalkyl, (default-mg/kg) R’ is H, C, to C, alkyl, or substituted C, to C, alkyl; ICso for uterine weight: Concentration of compound that R is H, OH, NH, C, to C alkyl, substituted C to C, gives half-maximal decrease in ECso progesterone alkyl, C. to Calkenyl, Substituted C to Calkenyl, induced decidual response (default-mg/kg) or COR; All publications cited in this Specification are incorpo 60 R is H, C, to C alkyl, substituted C to C alkyl, aryl, rated herein by reference herein. While the invention has Substituted aryl, C to Calkoxy, Substituted C to C. been described with reference to a particularly preferred alkoxy, C to C aminoalkyl, or Substituted C to C. embodiment, it will be appreciated that modifications can be aminoalkyl, made without departing from the Spirit of the invention. 65 R" is selected from the group consisting of (iv) and (v): Such modifications are intended to fall within the scope of (iv) a Substituted benzene ring containing the Sub the appended claims. stituents X, Y and Z as shown below: US 6,399,593 B1 31 32 group consisting of H, halogen, CN, NO, C to C. alkyl, and C to C alkoxy, R is H or C, to C, alkyl. 3. The method according to claim 1, wherein the proges rá 5 N tational agent is levonorgestrel and wherein: R'=R and are selected from the group consisting of C, to C. alkyl and substituted C, to C, alkyl, or R' and R' are fused to form the carbon-based Saturated 3 to 6 X is Selected from the group consisting of halogen, CN, membered Spirocyclic ring, C to C alkyl, Substituted C to C alkyl, C to C. R is H, OH, NH, C, to C, alkyl, substituted C, to C, alkoxy, Substituted C to C alkoxy, C to C alkyl, or COR; thioalkoxy, Substituted C to C thioalkoxy, amino, C to C aminoalkyl, Substituted C to C aminoalkyl, R is H, C, to C, alkyl, or C, to C alkoxy; NO, C to C perfluoroalkyl, 5 or 6 membered het 15 R" is selected from the group consisting of (viii), (ix), and erocyclic ring containing in its backbone 1 to 3 (x): heteroatoms, CORP, OCORP, and NRCORP; (viii) the Substituted benzene ring (iv) of the formula: R’ is H, C, to C, alkyl, substituted C, to C, alkyl, aryl, Substituted aryl, C to C alkoxy, Substituted C to C. alkoxy, C to C aminoalkyl, or Substituted C to C. aminoalkyl, R is H, C,1. to C, alalkyl, Ky or substituted C,1. to C, alalkyl; Ky Y and Z are independent substituents selected from the group consisting of H, halogen, CN, NO, C to C. 25 alkoxy, C to C alkyl, and C to C thioalkoxy; and (v) a five or six membered ring having in its back wherein bone 1, 2, or 3 heteroatoms selected from the X is Selected from the group consisting of halogen, group consisting of O, S, SO, SO and NR and CN, C to C alkoxy, C to C alkyl, NO, C to containing one or two independent Substituents C. perfluoroalkyl, 5 membered heterocyclic ring Selected from the group consisting of H, halogen, containing in its backbone 1 to 3 heteroatoms, and CN, NO, C, to C alkyl, C, to C alkoxy, C, to C to C thioalkoxy, Caminoalkyl, COR, and NRCOR; Y is on the 4 or 5' position and is selected from the R is H, C, to C, alkyl, substituted C, to C, alkyl, aryl, group consisting of H, halogen, CN, NO, C to Substituted aryl, C to C alkoxy, Substituted C to C. 35 Calkoxy, C to C alkyl, and C to C thioalkoxy, alkoxy, C to C aminoalkyl, or Substituted C to C. (ix) the five or six membered ring (v) of the structure: aminoalkyl, R’ is H, C,1. to C alKyalkyl, or substituted C,1. to C alKyalkyl; X R is H or C, to C, alkyl; 40 W is O or a chemical bond; or a pharmaceutically acceptable Salt thereof; and c) optionally, a third phase of daily dosage units of an orally and pharmaceutically acceptable placebo for the remaining days of the 28 consecutive days in which no 45 U is O, S, or NR; antiprogestin, progestin or estrogen is administered; X is Selected from the group consisting of halogen, wherein the total daily dosage units of the first, Second CN, NO, C to C alkyl, and C to C alkoxy; and third phases equals 28. Y is selected from the group consisting of H and C 2. The method according to claim 1, wherein the proges to C alkyl, and tational agent is levonorgestrel and wherein: 50 (x) the five or six membered ring (v) of the structure: R" is H, C, to C alkyl, substituted C to C alkyl, C. to Cs cycloalkyl, Substituted C to Cs cycloalkyl, aryl, Substituted aryl, heterocyclic, Substituted heterocyclic, COR, or NRCOR; 55 R" is (vi) or (vii): (vi) the substituted benzene ring (iv), wherein X is Selected from the group consisting of halogen, CN, X is N or CX; C to C alkyl, Substituted C to C alkyl, C to C. X is halogen, CN or NO. alkoxy, Substituted C to C alkoxy, C to C. 60 4. The method according to claim 1, wherein the proges thioalkoxy, Substituted C to C thioalkoxy, C to C. tational agent is levonorgestrel and wherein: aminoalkyl, Substituted C to C aminoalkyl, NO, C to C perfluoroalkyl, a 5 membered heterocyclic R=R and are CH or R' and R are fused to form the ring containing in its backbone 1 to 3 heteroatoms, carbon-based Saturated 6 membered Spirocyclic ring, CORP, OCORP, and NRCORP; or 65 R is H, OH, NH, CH, substituted CH, or COR; (vii) the five or six membered ring (V), wherein said one R" is the substituted benzene ring (iv) having the structure or two independent Substituents are Selected from the shown below: US 6,399,593 B1 34

I R1 R2 R A. n W

NellsN N O

1O k wherein: X is halogen, CN, methoxy, or NO; wherein: Y is on the 4' or 5" position and is selected from the A, B and D are N or CH, with the proviso that A, B and group consisting of H and halogen. D can not all be CH; 5. The method according to claim 1, wherein the proges 15 R" is selected from the group consisting of H, C, to C. tational agent is levonorgestrel and wherein: alkyl, Substituted C to C alkyl, C. to Cs cycloalkyl, Substituted C to Cs cycloalkyl, aryl, Substituted aryl, R=R and are CH or R and R are fused to form the heterocyclic, substituted heterocyclic, COR, and carbon-based Saturated 6 membered Spirocyclic ring, NRFCOR^; R is H, OH, NH, CH, substituted CH, or COR; R is selected from the group consisting of H, C, to C. alkyl, Substituted C to C alkyl, C. to C alkenyl, R" is the five or six membered ring (v) of the structure: Substituted C to C alkenyl, Cs to Cs cycloalkyl, Substituted C to Cs cycloalkyl, aryl, Substituted aryl, X heterocyclic, substituted heterocyclic, COR, and NRFCOR^; or R' and R are fused to form a spirocyclic ring Selected from the group consisting of (i), (ii), and (iii): (i) a carbon-based Saturated 3 to 8 membered Spiro 3O cyclic ring; U is O, S, or NH; (ii) a carbon-based 3 to 8 membered Spirocyclic ring X is halogen, CN, or NO; having one or more carbon-carbon double bonds, Y is H or C to C alkyl. and 6. The method according to claim 1 wherein the anti (iii) a 3 to 8 membered heterocyclic ring containing progestin compound is 6-(3-Chloro-phenyl)-4,4-dimethyl-1, 35 in its backbone one to three heteroatoms Selected from the group consisting of O, S and N, 4-dihydro-3-Oxa-1,8-diaza-naphthalene-2-one. the Spirocyclic rings of (i), (ii), and (iii) being option 7. The method according to claim 1 wherein the proges ally substituted by from 1 to 4 groups selected from tational agent is Selected from the group consisting of the group consisting of fluorine, C to C alkyl, C to levonorgestrel, norgestrel, desogestrel, 3-ketodesogestrel, 40 C alkoxy, C to C thioalkyl, CF, OH, CN, NH, NH(C. to C alkyl), and N(C. to C alkyl); nore thindrone, gestodene, no rethindrone acetate, R is H, C, to C, alkyl, substituted C, to C alkyl, aryl, norgestimate, OSaterone, , trimegestone, Substituted aryl, C to Calkoxy, Substituted C to C. dienogest, drospire none, nomegestrol, and (17-deacetyl) alkoxy, C to C aminoalkyl, or Substituted C to C. norgestimate. 45 aminoalkyl, 8. The method according to claim 1 which comprises: R’ is H, C, to C, alkyl, or substituted C, to C, alkyl; R is H, OH, NH, C, to C alkyl, substituted C to C, a) a first phase of 21 daily dosage units of Said proges alkyl, C. to Calkenyl, Substituted C to Calkenyl, tational agent, or COR; b) a Second phase of 3 daily dosage units of Said anti 50 R is H, C, to C alkyl, substituted C to C alkyl, aryl, progestin compound of formula I; and Substituted aryl, C to Calkoxy, Substituted C to C. alkoxy, C to Caminoalkyl, or Substituted C to C. c) optionally, 4 daily dosage units of Said orally and aminoalkyl, pharmaceutically acceptable placebo to be adminis R" is selected from the group consisting of (iv) and (v): tered on each day of the 28-day cycle following the first 55 (iv) a Substituted benzene ring containing the Sub phase and Second phase. stituents X, Y and Z as shown below: 9. A method of contraception which comprises adminis tering to a female of child bearing age for 28 consecutive Z. days: 60 a) a first phase of from 14 to 24 daily dosage units of a er/ S. progestational agent equal in progestational activity to about 35 to about 100 lug levonorgestrel; b) a second phase of from 1 to 11 daily dosage units, at 65 a daily dosage of from about 2 to 50 mg, of an X is Selected from the group consisting of antiprogestin compound of Formula I: halogen, CN, C to C alkyl, Substituted C to US 6,399,593 B1 35 36 C alkyl, C to C alkoxy, Substituted C to C. R" is selected from the group consisting of (viii), (ix), and alkoxy, C to C thioalkoxy, Substituted C to (x): C. thioalkoxy, C to C aminoalkyl, Substi (viii) the Substituted benzene ring (iv) of the formula: tuted C to C aminoalkyl, NO, C to C. perfluoroalkyl, 5 membered heterocyclic ring containing in its backbone 1 to 3 heteroatoms, CORD, OCORD, and NRFCOR; R’ is H, C to C alkyl, substituted C to C. alkyl, aryl, Substituted aryl, C to C alkoxy, 1O Substituted C to C alkoxy, C to C aminoalkyl, or Substituted C to C ami noalkyl, wherein R is H, C, to C alkyl, or substituted C to C. X is Selected from the group consisting of halogen, alkyl, 15 CN, C to C alkoxy, C to C alkyl, NO, C to C. perfluoroalkyl, 5 membered heterocyclic ring Y and Z are independent Substituents Selected containing in its backbone 1 to 3 heteroatoms, and from the group consisting of H, halogen, CN, C to C thioalkoxy, NO, C to C alkoxy, C to C alkyl, and C Y is on the 4 or 5" position and is selected from the to C thioalkoxy; and group consisting of H, halogen, CN, NO, C to Calkoxy, C to C alkyl, and C to C thioalkoxy, (v) a five or six membered ring having in its back (ix) the five or six membered ring (v) of the structure: bone 1, 2, or 3 heteroatoms selected from the group consisting of O, S, SO, SO and NR and X containing one or two independent Substituents 25 Selected from the group consisting of H, halogen, CN, NO, C to C alkyl, and C to C alkoxy; R is H or C, to C, alkyl; W is O or a chemical bond; or a pharmaceutically acceptable Salt thereof, and U is O, S, or NR; c) optionally, a third phase of daily dosage units of an X is Selected from the group consisting of halogen, CN, NO, C to C alkyl and C to C alkoxy; orally and pharmaceutically acceptable placebo for the Y' is Selected from the group consisting of H and C remaining days of the 28 consecutive days in which no 35 to C alkyl, and antiprogestin, progestin or estrogen is administered; (x) the five or six membered ring (v) of the structure: wherein the total daily dosage units of the first, Second and third phases equals 28. 10. The method according to claim 9 wherein the proges tational agent is levonorgestrel wherein: 40 when R is: (vi) the substituted benzene ring (iv), X is selected from the group consisting of halogen, CN, C to C alkyl, X is N or CX; 45 X is halogen, CN or NO. Substituted C to C alkyl C to C alkoxy, Substi 12. The method according to claim 9, wherein the proges tuted C to C alkoxy, C to C thioalkoxy, Substi tational agent is levonorgestrel and wherein: tuted C to C thioalkoxy, C to C aminoalkyl, R=R and are CH; Substituted C to C aminoalkyl, NO, C to C. R is H, OH, NH, CH, substituted CH, or COR; perfluoroalkyl, a 5 membered heterocyclic ring con 50 R" is the substituted benzene ring (iv) having the structure taining in its backbone 1 to 3 heteroatoms, CORP, shown below: OCORP, and NRFCORP; or (vii) the five or six membered ring (V), Said one or two independent Substituents are Selected from the group consisting of H, halogen, CN, NO, C, to C alkyl, 55 and C to C alkoxy, R is H or C, to C, alkyl. 11. The method according to claim 9, wherein the proges tational agent is levonorgestrel and wherein 60 R=R and are selected from the group consisting of C, to wherein: C alkyl and Substituted C to C alkyl, X is halogen, CN, methoxy, or NO; Y is on the 4' or 5" position and is selected from the R is H, OH, NH, C, to C alkyl, Substituted C, to C, 65 group consisting of H and halogen. alkyl, or COR; 13. The method according to claim 9, wherein the proges R is H, C, to C, alkyl, or C, to C, alkoxy; tational agent is levonorgestrel and wherein: US 6,399,593 B1 37 38 R=R and are CH; wherein: R is H, OH, NH, CH, substituted CH, or COR; A, B and D are N or CH, with the proviso that A, B and R" is the five or six membered ring (v) of the structure: D can not all be CH; R'=R and are C, to C, alkyl or substituted C, to C, X alkyl or R and Rare fused to form a carbon-based 3 to 6 membered Saturated Spirocyclic ring; R is H, OH, NH, C, to C alkyl, substituted C to C. alkyl, or COR; 1O R is H, C, to C, alkyl, or C, to C, alkoxy; R" is selected from the group consisting of (i), (ii), and U is O, S, or NH; X is halogen, CN, or NO; (iii): Y is H or C to C alkyl. 15 (i) a Substituted benzene ring of the formula: 14. The method according to claim 9 wherein the anti progestin compound is a pharmaceutically acceptable Salt of 6-(3-Chloro-phenyl)-4,4-dimethyl-1,4-dihydro-3-oxa-1,8- diaza-naphthalen-2-one. 15. The method according to claim 9 wherein the proges tational agent is Selected from the group consisting of levonorgestrel, norgestrel, desogestrel, 3-ketodesogestrel, nore thindrone, gestodene, no rethindrone acetate, norgestimate, OSaterone, cyproterone acetate, trimegestone, 25 wherein dienogest, drospire none, nomegestrol, and (17-deacetyl) X is Selected from the group consisting of norgestimate. halogen, CN, C to C alkoxy, C to C alkyl, 16. The method according to claim 9, which comprises: NO, C, to C perfluoroalkyl, 5 membered a) a first phase of 21 daily dosage units of Said proges heterocyclic ring containing in its backbone 1 tational agent, to 3 heteroatoms, and C to C thioalkoxy, b) a Second phase of 3 daily dosage units of Said anti Y is on the 4' or 5" position and is selected from progestin compound of formula I; and the group consisting of H, halogen, CN, NO, c) optionally, 4 daily dosage units of Said orally and C to C alkoxy, C to C alkyl, and C to C. pharmaceutically acceptable placebo to be adminis 35 thioalkoxy, tered on each day of the 28-day cycle following the first (ii) a five membered ring of the structure: phase and Second phase. 17. The method according to claim 1, wherein R' and R' X are fused to form a Spirocyclic ring which is optionally 40 substituted by from 1 to 3 C, to C alkyl groups. 18. The method according to claim 9, wherein R and R' are fused to form a Spirocyclic ring which is optionally substituted by from 1 to 3 C to C alkyl groups. 45 19. A method of contraception, which comprises admin istering to a female of child bearing age for 28 consecutive U is O, S, or NR; days: X is Selected from the group consisting of halogen, CN, NO, C to C alkyl, C to C. a) a first phase of from 14 to 24 daily dosage units of a 50 perfluoroalkyl, 5-membered heterocyclic ring progestational agent equal in progestational activity to containing in its backbone 1 to 3 heteroatoms, about 35 to about 100 lug levonorgestrel; C to C thioalkoxy, and C to C alkoxy, b) a second phase of from 1 to 11 daily dosage units, at Y' is Selected from the group consisting of H, a daily dosage of from about 2 to 50 mg, of an halogen, CN, NO, C to C alkoxy, C to C. antiprogestin compound of Formula I: 55 thioalkoxy, and C to C alkyl, R is H or C, to C, alkyl; and R1 R2 (iii) a six membered ring of the structure: R A. n W 60

B N 2 N 1sO k 65 X is N or CX; X is halogen, CN or NO; US 6,399,593 B1 39 40 W is O or a chemical bond; or a pharmaceutically (i) a Substituted benzene ring of the formula: acceptable Salt thereof, and c) optionally, a third phase of daily dosage units of an orally and pharmaceutically acceptable placebo for the remaining days of the 28 consecutive days in which no antiprogestin, progestin or estrogen is administered; wherein the total daily dosage units of the first, Second and third phases equals 28. 20. A method of contraception which comprises admin 1O istering to a female of child bearing age for 28 consecutive wherein days, X is Selected from the group consisting of a) a first phase of from 14 to 24 daily dosage units of a halogen, CN, methoxy, NO, and 2-thiazole; progestational agent equal in progestational activity to 15 Y is on the 4' or 5" position and is selected from the group consisting of H and F; about 35 to about 100 lug levonorgestrel; (ii) a five membered ring of the structure: b) a second phase of from 1 to 11 daily dosage units, at a daily dosage of from about 2 to 50 mg, of an X antiprogestin compound of Formula I:

R1 R2 R A. 25 n W U is O, S, or NH; X is Selected from the group consisting of B N 2 N 1sO halogen, CN, or NO; l Y' is Selected from the group consisting of H and C to C alkyl, W is O or a chemical bond; or a pharmaceutically wherein: acceptable Salt thereof, and A, B and D are N or CH, with the proviso that A, B and c) optionally, a third phase of daily dosage units of an D can not all be CH; 35 orally and pharmaceutically acceptable placebo for the R=R and are CH or R' and R are fused to form a remaining days of the 28 consecutive days in which no carbon-based 3 to 6 membered Saturated Spirocyclic antiprogestin, progestin or estrogen is administered; ring: wherein the total daily dosage units of the first, Second R is H, OH, NH, CH, substituted CH, or COR; and third phases equals 28. R is H, C, to C, alkyl, or C, to C, alkoxy; 40 R" is selected from the group consisting of (i) and (ii): k k k k k UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OF CORRECTION

PATENT NO. : 6,399,593 B1 Page 1 of 1 DATED : April 19, 2002 INVENTOR(S) : G. Grubb et al.

It is certified that error appears in the above-identified patent and that said Letters Patent is hereby corrected as shown below:

Title page, Item 57, ABSTRACT, after the word “using”, delete “progestational agents and”. Column 17 Line 61, replace “thil” with -- thyl --. Column 26 Line 23, Table 4, replace "0.00' with -- 0.001 --. Line 43, replace "X-100II.” Start new paragraph with -- II. --. Column 35 Line 54, after “(v)', insert -- wherein --. Line 56, replace “NO” with -- NO --.

Signed and Sealed this Twenty-sixth Day of November, 2002

Attest.

JAMES E ROGAN Attesting Officer Director of the United States Patent and Trademark Office