AU2011244870B2 - Cyclic progestin regimens and kits - Google Patents

Abstract

Abstract A method of contraception is provided which involves delivery of 21 to 27 consecutive days of a progestin such as tranaproget in the absence of an estrogen or other steroidal compound, 5 followed by I to 7 days without an effective amount of an active agent. Also described is a pharmaceutically useful kit to facilitate delivery of this regimen.

Description

AUSTRALIA Patents Act COMPLETE SPECIFICATION (ORIGINAL) Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: Wyeth Actual Inventor(s): Ginger Dale Constantine, Gary Sondermann Grubb Address for Service and Correspondence: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: CYCLIC PROGESTIN REGIMENS AND KITS Our Ref: 926697 POF Code: 878/460161 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): -1- Ia CYCLIC PROGESTIN REGIMENS AND KITS BACKGROUND OF INVENTION The present application is a divisional application from Australian Patent Application 5 No. 2005269938; the entire disclosure of which is incorporated herein by reference. Soon after the introduction of progestin/estrogen combination oral contraceptives (OCs) pills in 1960, several progestin-only pills (POPs) were introduced. The dose of the progestin in the POPs was made lower than in the combined OCs to minimize the occurrence of amenorrhea resulting from complete ovarian suppression. Consequently, ovulation was 10 inhibited in about half the users of POPs. (The standard POPs primarily depend upon cervical mucus thickening to provide contraceptive protection for those who ovulate). Partly because of the lower progestin dose, the absence of exogenous estrogen and the absence of regular withdrawal bleed, POP users have a much higher rate of unscheduled breakthrough bleeding and spotting than combination OC users. Primarily because of the bleeding problems, POPs 15 are used by only about 1-2% of contracepting women, compared to about 30% using combination OCs. What is needed is a progestin-containing contraceptive which avoids breakthrough bleeding and spotting problems. 20 SUMMARY OF THE INVENTION The present invention provides a contraceptive regimen which involves delivery of an effective amount of a progestin for 21 to 27 consecutive days following by I to 7 consecutive days without delivery of same. In one embodiment, the progestin is 5-(4,4-Dimethyl-2-thioxo 1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yl)-1-methyl-iH-pyrrole-2-carbonitrile, also termed 25 tanaproget. The invention further provides a pharmaceutical kit for use in the delivery of the regimen. Other aspects of the invention will be readily apparent from the following detailed description of the invention. 30 C:\pcono SPEC-92097j1J.doc 2 DETAILED DESCRIPTION OF THE INVENTION In one aspect, the present invention provides a method of contraception in a female of child-bearing age. In this method, a progestin, or combination of progestins, is delivered for a period of consecutive days as the sole active (i.e., anti 5 contraceptive) agent in order to prevent conception. In another aspect, the present invention provides for the use of a progestin, or combination of progestins, in preparing a medicament useful for contraception in a female of child-bearing age. In one embodiment, the medicament comprises one phase in which, from 21 to 27 daily dosage units are consecutively administered, each 10 containing an active agent comprising a progestin. In a further embodiment, the medicament also comprises from I to 7 daily dosage units of a pharmaceutically acceptable placebo for administration in a second phase. In other embodiments, progestins are useful in preparing medicaments useful in any of the methods of contraception described herein. 15 Without wishing to be bound by theory, it is believed that this invention will reduce the bleeding problems of conventional progestin-only contraceptives in two ways. First, due to the incorporation of I to 7 days in each cycle with no effective amount of a progestin, a regular withdrawal bleed will occur approximately every 28 days. Second, increasing the progestin dose above standard progestin-only 20 contraceptives, produces almost complete ovulation inhibition and the composition of the invention does not depend on cervical mucus thickening to maintain. the contraceptive effectiveness as do conventional progestin-only regimens. The term "progestin," as used herein, refers to any progestationally active compound, i.e., any compound that binds to and activates the progesterone receptor. 25 Representative progestins include progesterone synthetic derivatives such as, for example, 17-hydroxy progesterone esters, 19-nor-I 7-hydroxy progesterone esters, 17a-ethinyltestosterone and derivatives thereof,. I 7ct-ethinyl-19-nor-testosterone and derivatives thereof, norethindrone, norethindrone acetate, ethynodiol diacetate, dydrogesterone, medroxy-progesterone acetate, norethynodrel, allylestrenol, 30 lynoestrenol, fuingestanol acetate, medrogestone, norgestrienone, dimethiderome, ethisterone, cyproterone acetate, levonorgestrel, dl-norgestrel, d-1 7a-acetoxy-1 30- 3 ethyl-I 7a -a-ethinyl-gon-4-en-3-one oxime, cyproterone acetate, gestodene, desogestrel, etonorgestrel, norgestimate and norelgestromin. Other compounds with progestational activity used in oral contraceptives include chlormadione, dienogest, and drospirenone. 5 In one desirable embodiment, the progestin is 5-(4,4-Dimethyl-2-thioxo-l,4 dihydro-2H-benzofd] [ ,3]oxazin-6-yl)-I-methyl-IH-pyrrole-2-carbonitrile, also termed tanaproget and NSP-989. This compound can have the formula: NC / / N 0

H

3 C N S H 10 and encompasses pharmaceutically acceptable salts, esters or other prodrug forms thereof This compound and methods of making same are described in US Patent No. 6,436,929, US Patent Application No. 11/113,794 (filed April 25, 2005), and US Provisional Patent Application Nos. 60/675,550 (filed April 28, 2005); 60/675,551 15 (filed-April 28,2005); 60/675,559 (filed April 28, 2005); 60/675,737 (filed April 28, 2005); and 60/675,738 (filed April 28, 2005). In addition, other progestins described in US Patent Nos. 6,436,929; 6,355,648; 6,521,657; 6,407,101; 6,562,857; and 6,358,947, US Patent Publication No. 2003-0158182-AI, and US Provisional Patent Application No. 60/601,254 (filed 20 August 13, 2004) may be useful in the methods and kits of the invention. The progestin compounds useful in the present invention can be used in the form of salts derived from pharmaceutically or physiologically acceptable acids or bases. These salts include, but are not limited to, the following salts with inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and, as the case 25 may be, such organic acids as acetic acid, oxalic acid, succinic acid, and maleic acid. Other salts include salts with alkali metals or alkaline earth metals, such as sodium, potassium, calcium or magnesium in the form of esters, carbamates and other - I conventional "pro-drug" forms, which, when administered in such form, convert to the active moiety in vivo. The method of the invention is performed for a period of time corresponding to the length of a menstrual cycle, i.e., in the range of 23 to 35 days, with 28 days 5 being the average. Thus, the method of the invention involves delivering a daily dosage unit containing an effective amount of an active agent consisting of a progestin to a female of child bearing age over a period of 18 to 28 consecutive days followed by I to 7 consecutive days in which no effective amount of an active agent is delivered to the subject. 10 The term "effective amount" of a progestin(s) is a dosage that provides contraception. Without being bound by theory, this is achieved primarily by preventing ovulation. The term "no effective amount" of a progestin(s) is used to refer to the I to 7 days following delivery of an effective amount of the progestin(s). During this period, preferably, no amount of a progestin(s) is delivered to the animal. 15 However, it is possible, that a sustained release formulation or other delivery method may be "leaky" and continue to deliver low amounts of a progestin which are not effective at contraception during this period. The phrase "no effective amount" encompasses delivery of no amount of progestin(s). According to the present invention, a female is preferably a human. However, 20 as used herein, a female can include non-human mammals, e.g., cattle or livestock, horses, pigs, domestic animals, etc. In one aspect, the method of invention involves delivering a daily dosage unit containing an active agent consecutively for at least 21 of 28 consecutive days. In the embodiment, the regimen consists of delivering a progestin to a female of child 25 bearing age over a period of 21 to 27 consecutive days followed by I to 7 consecutive days in which no effective amount or no amount of active agent is delivered to the subject. Optionally, the period of I to 7 days in which no effective amount of an active agent is delivered to the subject can involve delivery of a second phase of daily dosage units of I to 7 days of a pharmaceutically acceptable placebo. Alternatively, 30 during this "placebo period", no placebo is administered.

4a One preferred embodiment is a method of contraception in a female of child bearing age, which comprises administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit containing an effective amount of from 5 0.03 to 0.6 mg of an active agent comprising 5-(4,4-Dimethyl-2-thioxo-1,4-dihydro 2H-benzo[d][1,3]oxazin-6-yl)-1-methyl-iH-pyrrol-2-carbonitrile (Tanaproget) of formula: NC N 00 H3C N "S H ; and 0 (b) a second phase of from 1 to 7 days wherein no effective amount of an active agent is administered; wherein said method is performed in the absence of a second steroidal compound. 5 Another embodiment is a method of contraception in a female of child bearing age, said method comprising administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit containing an effective amount of from 20 0.03 to 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro 2H-benzo[d][1,3]oxazin-6-yl)-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of said progestin is administered; wherein said method is performed in the absence of an estrogen. 25 A further embodiment is a method of contraception in a female of child bearing age which consists essentially of administering over a period of 28 consecutive days: 4b (a) a first phase in which from 21 to 27 daily dosage units of from 0.3 to 0.6 mg of an active agent are administered, each daily dosage unit consisting of an effective amount of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6 5 yl]-1 -methyl-i H-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of Tanaproget is administered; wherein said method is performed in the absence of a second steroidal compound. 0 A further embodiment is a method of contraception in a female of child bearing age which consists essentially of administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of from 0.03 to 0.6 mg of an active agent are administered, each daily dosage unit consisting of an 5 effective amount of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6 yl]-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of Tanaproget is administered; wherein said method is performed in the absence of an estrogen. A further embodiment is a method of contraception in a female of child bearing age which consists of administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit consisting of an effective amount of 25 from 0.03 to 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4 dihydro-2H-benzo[d][1,3]oxazin-6-yl]-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of Tanaproget is administered; 30 wherein said method is performed in the absence of a second steroidal compound.

4c A further embodiment is a method of contraception in a female of child bearing age which consists of administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit consisting of an effective amount of 5 from 0.03 to 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4 dihydro-2H-benzo[d][1,3]oxazin-6-yl]-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of Tanaproget is administered; 0 wherein said method is performed in the absence of an estrogen.

In one embodiment, the method of the invention involves delivering a progestin as the sole active agent for 21 consecutive days followed by 7 days in which no effective amount of an active agent is delivered. Optionally, during these 7 days, a second phase of 7 daily dosage units of an orally and pharmaceutically acceptable 5 placebo can be delivered. In another embodiment, the method of the invention involves delivering a progestin as the sole active agent for 23 consecutive days followed by 5 days in which no effective amount of an active agent is delivered. Optionally, during these 5 days, a second phase of 5 daily dosage units of an orally and pharmaceutically acceptable 10 placebo can be delivered. In still another embodiment, the method of the invention involves delivering a progestin as the sole active agent for 25 consecutive days followed by 3 days in which no effective amount of an active agent is delivered. Optionally, during these 3 days, a second phase of 3 daily dosage units of an orally and pharmaceutically acceptable 15 placebo can be delivered. In still another embodiment, the method of the invention involves delivering a progestin as the sole active agent for 27 consecutive days followed by I day in which no effective amount of an active agent is delivered. Optionally, a second phase of I daily dosage unit of an orally and pharmaceutically acceptable placebo can be 20 delivered. This invention includes the use of pharmaceutical compositions containing one or more progestin compound(s) as the sole active ingredient in the formulation and regimen. The progestin compounds are formulated with a pharmaceutically acceptable carrier or excipient. 25 Suitably, the progestins used in the invention are formulated for delivery by any suitable route including, e.g., transdermal, mucosal (intranasal, buccal, vaginal), oral, parenteral, etc, by any suitable delivery device including, e.g., transdermal patches, topical creams or gels, a vaginal ring, among others. In a further embodiment, the progestins are delivered by any suitable route in a sustained release 30 formulation. Such sustained release formulations are known to those of skill in the art.

6 When the compounds are employed for the above utilities, they may be combined with one or more pharmaceutically acceptable carriers or excipients, for example, solvents, diluents and the like. When formulated for oral delivery, the progestin compound can be in theform of a tablet, capsule, caplet, gel tab, dispersible 5 powders, granules, or suspensions containing, for example, from about 0.05 to 5% of suspending agent, syrups containing, for example, from about 10 to 50% of sugar, and elixirs containing, for example, from about 20 to 50% ethanol, and the like. When formulated for parenteral delivery, the compositions can be delivered in the form of sterile injectable solutions or suspensions containing from about 0.05 to 5% 10 suspending agent in an isotonic medium. Such pharmaceutical preparations may contain, for example, from about 25 to about 90% of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight. The effective dosage of active ingredient employed may vary depending on the particular compound employed, the mode of administration and the degree of 15 ovarian suppression desired. However, in general, satisfactory results are obtained when the compounds of the invention are administered at a daily dosage of from about 0.03 to 0.6 mg, or about 0.1 to about 0.5 mg, preferably given daily or in a sustained release form. This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily 20 or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. These active compounds (one or more progestins) may be administered orally. Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, non-ionic surfactants, 25 ethanol (e.g., glycerol, propylene glycol and liquid polyethylene glycols), suitable mixtures thereof, and vegetable or edible oils such as com, peanut and sesame oils, as are appropriate to the nature of the active ingredient and the particular form of administration desired. Adjuvants customarily employed in the preparation of pharmaceutical compositions may be advantageously included, such as flavoring 30 agents, coloring agents, preserving agents, and antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA.

7 The preferred pharmaceutical compositions from the standpoint of ease of preparation and administration are solid compositions, particularly tablets and hard filled or liquid-filled capsules. Oral administration of the compounds is preferred. These active compounds may also be administered via a vaginal ring. 5 Suitably, use of the vaginal ring is timed to the 28 day cycle. In one embodiment, the ring is inserted into the vagina, and it remains in place for 3 weeks. During the fourth week, the vaginal ring is removed and menses occurs. The following week a new ring is inserted to be wom another 3 weeks until it is time for the next period. In another embodiment, the vaginal ring is inserted weekly, and is replaced for three consecutive 10 weeks. Then, following one week without the ring, a new ring is inserted to begin a new regimen. In yet another embodiment, the vaginal ring is inserted for longer, or shorter periods of time. For use in the vaginal ring, a progestin compound is formulated in a manner similar to that described for contraceptive compounds previously described for 15 delivery via a vaginal ring. See, e.g., US Patent Nos. 5,972,372; 6,126,958; and 6,125,850. Optionally, a progestin composition can be formulated for parenteral delivery in a sustained release formulation and administered by injection, e.g., monthly or quarterly. 20 In another aspect of the invention, a progestin compound is formulated for delivery via a cream or a gel, by a suitable route. Suitably, carriers for such routes are known to those of skill in the art. In still another aspect of the invention, the progestin compound(s) are delivered via a transdermal patch. Suitably, use of the patch is timed to the 28 day 25 cycle. In one embodiment, the patch is applied via a suitable adhesive on the skin; where it remains in place for 1 week and is replaced weekly for a total period of three weeks. During the fourth week, no patch is applied and menses occurs. The following week a new patch is applied to be worn to begin a new regimen. In yet another embodiment, the patch remains in place for longer, or shorter periods of time. 30 This invention also includes kits or packages of pharmaceutical formulations designed for use in the regimens described herein. Suitably, the kits contain one or 8 more progestin compounds as described herein. In one embodiment, the progestin is selected from among 17-hydroxy progesterone esters, 19-nor-I 7-hydroxy progesterone esters, 17c.-ethinyltestosterone and derivatives thereof, 17a-ethinyl-19 nor-testosterone and derivatives thereof, norethindrone, norethindrone acetate, 5 ethynodiol diacetate, dydrogesterone, medroxy-progesterone acetate, norethynodrel, allylestrenol, lynoestrenol, fuingestanol acetate, medrogestone, norgestrienone, dimethiderome, ethisterone, cyproterone acetate, levonorgestrel, di-norgestrel, d-I7a acetoxy-1 30-ethyl-1 7ca -a-ethinyl-gon-4-en-3-one oxime, cyproterone acetate, gestodene, desogestrel, etonorgestrel, norgestimate, norelgestromin, chlormadione, 10 dienogest, and drospirenone. In addition, other progestins described in US Patent Nos. 6,436,929; 6,355,648; 6,521,657; 6,407,101; and 6,562,857, may be useful in the methods and kits of the invention. In one desirable embodiment, the progestin is NSP-989, also termed tanaproget, or a pharmaceutically acceptable salt or prodrug thereof. 15 Advantageously, for use in the kits of the invention, the progestin is formulated for the desired delivery vehicle and route. For example, a progestin can be formulated for oral delivery, parenteral delivery, vaginal ring, transdernnal delivery, or mucosal delivery. In one embodiment,the kit of the invention is designed for daily oral 20 administration over a 28-day cycle, preferably for one oral administration per day, and organized so as to indicate a single oral formulation or combination of oral formulations to.be taken on each day of the 28-day cycle. Preferably each kit will include oral tablets to be taken on each the days specified; preferably one oral tablet will contain each of the combined daily dosages indicated. For example, a kit of the 25 invention can contain 21 to 27 daily dosage units of an effective amount of an active agent and, optionally, I to 7 daily dosage units of a placebo and other appropriate components including, e.g., instructions for use. The kit of the invention is preferably a pack (e.g. a blister pack) containing daily doses arranged in the order in which they are to be taken. 30 In another embodiment, the kit of the invention is designed for weekly or monthly administration via a vaginal ring over a 28-day cycle. Suitably, such a kit 9 contains individual packaging for each of the vaginal rings, i.e. one to three, required for a monthly cycle and other appropriate components, including, e.g., instructions for use. In another embodiment, the kit of the invention is designed for weekly or 5 monthly administration via a transdermal patch over a 28-day cycle. Suitably, such a kit contains individual packaging for each of the patches, i.e. one to three, required for a monthly cycle and other appropriate components including, e.g., instructions for use. In still another embodiment, the kit of the invention is designed for parenteral 10 delivery of the progestin. Such a kit is typically designed for delivery, at home and may include needles, syringes, and other appropriate packaging and instructions for use. In yet another embodiment, the kit of the invention contains a progestin compound in a gel or cream formulation. Optionally, the kit can include appropriate 15 packaging such as a tube or other container, an applicator, and/or instructions for use. In each of the regimens and kits described herein, it is preferred that the daily dosage of each pharmaceutically active component of the regimen remain fixed in each particular phase in which it is administered. It is also understood that the daily dose units described are to be administered in the order described, with the first phase 20 followed in order by the optional second phase. To help facilitate compliance with each regimen, it is also preferred that the kits contain the placebo described for the final days of the cycle. It is further preferred that each package or kit comprise a pharmaceutically acceptable package having indicators for each day of the 28-day cycle, such as a labeled blister package, dial dispenser, or other packages known in 25 the art. These dosage regimens may be adjusted to provide the optimal contraceptive effect. For example, several divided doses of each component may be administered daily or the dose may be proportionally increased or reduced as indicated by the contraceptive effectiveness. In the descriptions herein, reference to a daily dosage 30 unit may also include divided units which are administered over the course of each day of the cycle contemplated.

10 The following examples are illustrative only and are not intended to be a limitation on the present invention. Example 1 - Ovulation Inhibition by the Progestin Compound NSP-989 5 The activity of NSP-989 was evaluated orally in three 3 different rat models for progestin activity along with reference progestins medroxyprogesterone acetate (MPA) and trimegestone (TMG) in 2% Tween 80/0.5% methylcellulose vehicle. These models are described in Parts A, B and C of this example. A. Effect of NSP-989 in the Rat Ovulation Inhibition Model 10 The ovulation inhibition assay measures a compound's ability to inhibit ovulation in adult female rats. This activity is essential for contraceptive efficacy. In this assay, NSP-989 had a mean ED 1 00 value of 0.03 mg/kg, whereas both TMG and MPA had ED 1 00 values of I mg/kg (n = 2- to 3). Random cycling mature female Sprague-Dawley rats (-200 g) were 15 obtained from Charles River Laboratory (Boston, MA). Rats were synchronized for estrus with 2 sg of LHRH (in phosphate buffered saline containing 0.1% bovine serum albumin) administered subcutaneously (sc) per rat at 0900 h and again at 1600 h. Animals were allowed to rest for 8 days before the administration of test compounds. Animals were then grouped, with 7 to 9 rats per treatment group. The 20 morning of the ninth day following LHRH treatment, the rats were treated with test compounds once daily, by gavage. This continued for 4 consecutive days. The animals were euthanized the morning following the last treatment. Oviducts were removed, placed between 2 glass slides, and viewed through a dissecting microscope to count ova. The number of animals presenting ova in the oviduct from each 25 treatment group and the number of ova in the oviduct of each animal were recorded. B. Rat decidualization Decidualization Model The second rat model to determine progestational activity is the uterine decidualization assay in adult ovariectomized rats. Only compounds that are progesterone receptor agonists will be active in this model, as a progestin is absolutely 30 required to transform uterine stromal cells to differentiated decidual cells.

1 Rat decidualization assay was run as described previously [Lundeen SG, et al., "Rat uterine complement C3 expression as a model for progesterone receptor modulators: characterization of the new progestin trimegestone", J Steroid BiolMolBiol. 2001;78:137-143.] 5 Briefly, mature female Sprague-Dawley rats (-220 g) were ovariectomized at least 10 days prior tobefore treatment to reduce circulating sex. steroids. NSP-989 was administered once daily for seven 7 days orally by gavage (0.5 ml) in 2% Tween 80/0.5% methyl- cellulose vehicle. Approximately 24 hours after the third daily treatment, decidualization was induced in onel uterine horn of 10 each anesthetized rat by scratching the antimesometrial luminal epithelium with a blunt 21- gauge needle. The contralateral hom was not scratched and served as a non-stimulated control. Animals were euthanized by CO 2 asphyxiation 24 hoursr following the final treatment. The uteri were removed, and trimmed of fat, and the decidualized (D) and control (C) horns were weighed separately. The decidual 15 response is expressed as D/C. NSP-989 induced endometrial decidualization with an EDso value of 0.01 mg/kg (n = 23) and was approximately 40- and 100-fold more potent than MPA and TMG, respectively (Table 1). Table 1. Summary of Progestational Activity of NSP-989 in Various Rodent Models Rat Decidual Rabbit Clauberg Rat Ovulation Assay Rat C3 Assay Assay inhibition Assay (ED 5 o, mg/kg., (EDso, mg/kg. (AED5e, mg/kg, Compound

(ED

1 6, mg/kg, po) Po) po) Po) NSP-989 0.03 0.01 0.0005 0.001 MPA 1.0 0.4 0.03 0.03 TMG 1.0 1.0 0.005 0.001 20 C. Rat Uterine C3 model The third model for PR agonist activity was the adult ovariectomized rat uterine C3 model. This assay evaluates the ability of a progestin to block estrogen-induced C3 expression in the uterine epithelium. Ovariectomized female, 60 day-old Sprague-Dawley rats were 25 obtained from Ha'rlan (Indianapolis, IN). Ovariectomies were performed by the supplier a minimum of 8 days prior tobefore treatment. The rats were randomized 12 and placed in groups of 6. The animals were treated once daily for (2) two days orally by gavage (p.o.) in a volume of 0.5 mL. On the second day of treatment, the animals were also treated with EE (0.08 mg/kg body weight (BW)) orally by gavage. Approximately 24 hours after the final treatment, the animals were euthanized by C02 5 asphyxiation. The uteri were then removed, stripped of remaining fat and mesentery, weighed, and snap-frozen on dry ice. Total RNA was isolated from the uteri using the Trizol Reagent (GibcoBRL) as described by the manufacturer.. Real-time reverse transcription polymerase chain reaction (RT-PCR) as previously reported [Sampath D, et al., "Aberrant expression of Cyr61, a member of the CCN 10 (CTGF/Cyr6l/CeflO/NOVH) family, and dysregulation by 17b-estradiol and basic fibroblast growth factor in human uterine leiomyomas" J Clin Endocrinol Metab. 2001;86:1707-1715] was used to quantitate complement C3 expression. Briefly, RNA samples were DNAse-I treated using a DNA-free kit (Ambion). A total of 50 ng of RNA was analyzed in triplicate using C3 specific primer pair (5'primer 15 GGTCGGTCAAGGTCTACTCCTACTA [SEQ ID NO: 1], 3'primer CACAGCGGCACATTCATTG [SEQ ID NO: 2]) and customized probe (6FAM AGCATrCCATCGTCCTTCTCCGGATG-TAMRA [SEQ ID NO: 3]). C3 messenger RNA (mRNA) levels were normalized to 18s 18S ribosomal RNA contained within each sample reaction using primers and probe supplied by PE 20 Applied Biosystems. The mean EDso value for NSP-989 was 0.0005 mg/kg (n =6). Both MPA and TMG had mean EDSO values of 0.03 and 0.005 mg/kg, respectively in this assay. In summary, in the rats, in several unrelated models for progestin activity, NSP-989 was 3010- to 10060- fold more potent than the reference progestins used in 25 these studies. D. Rabbit Endometrial Transformation (Clauberg) Assay In addition to the rat progestational models described above, NSP-989 was also evaluated in the Clauberg model, a classic progestational assay in the rabbit endometrial transformation model [McPhail MK, "The assay of progestin." JPhysiol. 30 1934; 83:145-1567]. Briefly, immature female New Zealand White rabbits (~1 kg body weight) were injected subcutaneously with 5 pg 170-estradiol (E 2 )/rabbit/day for 13 six consecutive days. Beginning 24 hours after the final E 2 injection, vehicle alone or test compounds were given orally for (5) five consecutive days. Progestational activity was determined by increases in uterine weight and endometrial glandular arborization (McPhail Index). 5 In limited dose response studies, NSP-989 had an estimated ED 5 0 (AEDso) of 0.001 mg(kg. Its potency in this assay was similar to TMG and about 30 fold more potent than MPA. Example 2 - Cyclic Regimen Using Progestin Compound 10 A phase 2, randomized, double-blind, multicenter, dose-ranging study of 3 doses of NSP-989 in a 21-day regimen followed by 7 days of placebo pills, and a comparator (the combination steroidal OC desogestrel (DSG) 150 sg/20 pg ethinyl estradiol for 21 days followed by 2 days of placebo pills, followed by 5 days of 10 pg EE, marketed in the United States under the name Mircette) is planned. 15 Approximately 20 sites will participate with approximately 16 subjects per site. However, the enrollment will be competitive, and additional subjects can be enrolled at any site. The study will have 2 parts. Part 1 (days 1-84) of the study will evaluate the ability of NSP-989 to produce ovarian suppression, along with evaluating cycle 20 control, side effects, and metabolic data. Part 2 (days 85-168).will continue to follow the subjects to collect cycle control, side effects, and metabolic data. The study will be monitored routinely by the blinded project medical monitor and study team for efficacy failures and safety Each subject will participate for up to 9 months, depending on the length of the subject's screening period. Eight (8) cycles will be 25 observed. The first cycle will be a baseline observation of ovulation. Six (6) treatment cycles will be followed by 1 posttreatment observation cycle to assess return to ovulation. The subjects will be healthy women of 2 18 years of age who are younger than 36 years at the time of randomization. Subjects must have had spontaneous regular (24- to 32-day) menstrual cycles for the 3-month period 30 preceding entry into the pretreatment observation cycle, excluding postabortal and nonbreastfeeding postpartum subjects. Postabortal and nonbreastfeeding postpartum 14 subjects must have completed at least 1 regular (24- to 32-day) spontaneous menstrual cycle before entry into the pretreatment observation cycle. The pretreatment observation cycle for all subjects will begin on.day I of the subsequent spontaneous menses after completion of the prestudy screening (visit 1). 5 The pretreatment observation cycle is a control cycle; no test article will be administered. Each subject will begin test article on the first day of her menstrual bleeding (first subject pack only). Each subject pack will contain NSP-989 or the steroid combination OC comparatQr. Subjects will take NSP-989 orally, once daily for 21 days (days 1 through 21), followed by 7 days of placebo pills (days 22 through 10 28) for 6 cycles. Subjects assigned to a steroid combination OC comparator, DSG 150 Mg, will take test article orally once daily for 21 days (days I through 21), followed by 2 days of placebo pills (days 22 through 23), followed by 5 days of 10 pg EE (days 24 through 28) for 6 cycles. There will also be a posttreatment cycle in which no test article will be administered and return to ovulation will be assessed. 15 Each subject will be randomly assigned to receive one of the following: Group Treatment A 100 pg of NSP-989 for 21 days followed by 7 days of.placebo pills B 200 Mg of NSP-989 for 21 days followed by 7 days of placebo pills C 300 pg of NSP-989 for 21 days followed by 7 days of placebo pills 20 D Desogestrel 150 pig for 21 days followed by 2 days of placebo pills, followed by 5 days of 10 pg EE Each subject will begin test article on the first day of her menstrual bleeding (first subject pack only). Subjects will take test article orally, once daily for 28 days, at approximately the same time each day. All subsequent subject packs will begin 25 following day 28 of the previous pill pack. Subjects will take test article daily without interruption during the treatment cycles. It is anticipated that one or more treatment groups A, B and C receiving a regimen of the invention will have experience effective contraception, cessation of ovulation, and all groups will have a withdrawal bleed during the fourth week of each 30 month of treatment.

15 Example 3 A blister pack with 28 blister containers is made with a cardboard, paperboard, foil or plastic backing and enclosed in a suitable cover. The blister containers are arranged to house a 5 sequence of 21 pills each providing a daily dose of 100 ptg of NSP-989 followed by 7 daily doses of placebo pills (or 7 empty blisters). Each blister container may conveniently be numbered or otherwise marked, e.g., starting with the first of the 21 dosage units that contain the active ingredient followed by 7 empty blisters or by 7 dosage units that contain no active agent. 10 Throughout the description and claims of the specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps. 15 A reference herein to a patent document or other matter which is given as prior art is not to be taken as an admission that that document or matter was known or that the information it contains was part of the common general knowledge as at the priority date of any of the claims. SPEC-7903osa0c

Claims (18)

1. A method of contraception in a female of child bearing age, which comprises administering over a period of 28 consecutive days: 5 (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit containing an effective amount of from 0.03 to 0.6 mg of an active agent comprising 5-(4,4-Dimethyl-2-thioxo-1,4-dihydro 2H-benzo[d][1,3]oxazin-6-yl)-1-methyl-iH-pyrrol-2-carbonitrile (Tanaproget) of formula: 0 NC N0 H3C N -"S H ;and (b) a second phase of from 1 to 7 days wherein no effective amount of an active agent is administered; 5 wherein said method is performed in the absence of a second steroidal compound.

2. The method according to claim 1, wherein the effective amount is between 0.1 and 0.5 mg. 20

3. The method according to claim 1 or claim 2, wherein the effective amount is 0.1 mg.

4. The method according to claim 1 or claim 2, wherein the effective amount is 25 0.2 mg.

5. The method according to claim 1 or claim 2, wherein the effective amount is 0.3 mg. 17

6. The method according to any one of claims 1 to 5, wherein from 1 to 7 daily dosage units of a pharmaceutically acceptable placebo are delivered in the second phase.

7. The method according to any one of claims 1 to 5, which comprises: 5 a) a first phase of 21 daily dosage units; and b) a second phase of 7 daily dosage units of an orally and pharmaceutically acceptable placebo.

8. The method according to any one of claims 1 to 5, which comprises: 0 a) a first phase of 23 daily dosage units; and b) a second phase of 5 daily dosage units of an orally and pharmaceutically acceptable placebo.

9. The method according to any one of claims 1 to 5, which comprises: 5 a) a first phase of 25 daily dosage units; and b) a second phase of 3 daily dosage units of an orally and pharmaceutically acceptable placebo.

10. The method according to any one of claims 1 to 5, which comprises: to a) a first phase of 27 daily dosage units; and b) a second phase of 1 daily dosage units of an orally and pharmaceutically acceptable placebo.

11. The method according to claim 1, which produces complete ovulation 25 inhibition in said female.

12. The method according to claim 1, which does not rely on cervical mucus thickening of said female. 30

13. A method of contraception in a female of child bearing age, said method comprising administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit containing an effective amount of from 0.03 to 18 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H benzo[d][1,3]oxazin-6-yl)-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of said progestin is administered; 5 wherein said method is performed in the absence of an estrogen.

14. A method of contraception in a female of child bearing age which consists essentially of administering over a period of 28 consecutive days: (a) a first phase in which from 21 to 27 daily dosage units of from 0.3 to 10 0.6 mg of an active agent are administered, each daily dosage unit consisting of an effective amount of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yl]-1-methyl-iH pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of Tanaproget is administered; 15 wherein said method is performed in the absence of a second steroidal compound.

15. A method of contraception in a female of child bearing age which consists essentially of administering over a period of 28 consecutive days: ?0 (a) a first phase in which from 21 to 27 daily dosage units of from 0.03 to 0.6 mg of an active agent are administered, each daily dosage unit consisting of an effective amount of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yl]-1-methyl-iH pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of 25 Tanaproget is administered; wherein said method is performed in the absence of an estrogen.

16. A method of contraception in a female of child bearing age which consists of administering over a period of 28 consecutive days: 30 (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit consisting of an effective amount of from 0.03 to 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H benzo[d][1,3]oxazin-6-yl]-1-methyl-iH-pyrrole-2-carbonitrile (Tanaproget); and 19 (b) a second phase of from I to 7 days wherein no effective amount of Tanaproget is administered; wherein said method is performed in the absence of a second steroidal compound. 5

17. A method of contraception in a female of child bearing age which consists of administering over a period of 28 consecutive days: A method of contraception in a female of child bearing age which consists essentially of administering over a period of 28 consecutive days: 10 (a) a first phase in which from 21 to 27 daily dosage units of an active agent are administered, each daily dosage unit consisting of an effective amount of from 0.03 to 0.6 mg of an active agent consisting of 5-(4,4-dimethyl-2-thioxo-1,4-dihydro-2H benzo[d][1,3]oxazin-6-yl]-1-methyl-1H-pyrrole-2-carbonitrile (Tanaproget); and (b) a second phase of from 1 to 7 days wherein no effective amount of 15 Tanaproget is administered; wherein said method is performed in the absence of an estrogen.

18. The method according to any one of claims I and 13 to 17, substantially as hereinbefore described. ?O 2011244870 28 Oct 2011