WO2014143617A1

WO2014143617A1 - Deuterium-enriched tanaproget and processes for its preparation

Info

Publication number: WO2014143617A1
Application number: PCT/US2014/023256
Authority: WO
Inventors: Philmore Robertson, Jr.; Roberta Volonte; Augusto Canavesi
Original assignee: Teva Women's Health, Inc.
Priority date: 2013-03-14
Filing date: 2014-03-11
Publication date: 2014-09-18

Abstract

The present invention provides a deuterium enriched Tanaproget compound, isomer thereof, or a pharmaceutically acceptable salt thereof, including the following compound. These compounds may be utilized in reference standards and compositions. Also provided are methods of contraception, hormone replacement therapy, and treating and/or preventing uterine fibroids, dysfunctional bleeding, uterine leiomyomata, endometriosis, polycystic ovary syndrome, and benign and malignant neoplastic disease using these compounds.

Description

DEUTERIUM-ENRICHED TANAPROGET AND PROCESSES FOR ITS

PREPARATION

BACKGROUND OF THE INVENTION

[0001] Reference standards are utilized over a wide range of industries, including the pharmaceutical industries. A drug reference standard is a standardized

pharmaceutical compound which is used as a measurement base for similar compounds, i.e., a control or internal standard. A reference standard may be compared with other compounds, known or unknown, to determine the identity, strength, quality, and purity of other chemical compounds, i.e., pharmaceutical agents, biologies, etc. The comparisons may be biological, chemical, or spectroscopic comparisons, among others. Typically, a reference standard is a highly-characterized chemical compound. Reference standards may also be utilized to ensure proper performance of analytical instruments. Specifically, reference standards can be utilized to ensure that the analytical instrument is properly calibrated.

[0002] Progesterone receptor (PR) modulators are utilized in today's society for birth control compositions, emergency contraception, and hormone replacement therapy, among others. Tanaproget, 5-(4,4-dimethyl-2-thioxo-l,4-dihydro-2H-3, l- benzoxazin-6-yl)- 1 -methyl- lH-pyrrole-2-carbonitrile, is a progesterone receptor modulator and is effective in contraception, hormone replacement therapy, and treating carcinomas and adenocarcinomas, dysfunctional bleeding, uterine

leiomyomata, endometriosis, and polycystic ovary syndrome, among others.

Tanaproget

[0003] What is needed are alternate forms of Tanaproget for analytical, diagnostic, and treatment methods. SUMMARY OF THE INVENTION

[0004] In one aspect, a deuterium enriched Tanaproget compound, isomer thereof, or a pharmaceutically acceptable salt thereof is provided.

[0005] In a further aspect, a deuterium enriched Tanaproget compound is provided and is:

[0006] In another aspect, a reference standard is provided and contains a deuterium enriched Tanaproget compound.

[0007] In yet a further aspect, a composition is provided and contains a deuterium enriched Tanaproget compound and a pharmaceutically acceptable carrier.

[0008] In still another aspect, a method of contraception is provided and the method includes administering a deuterium enriched Tanaproget compound to a female in need thereof.

[0009] In a further aspect, a method of hormone replacement therapy is provided and the method includes administering a deuterium enriched Tanaproget compound to a subject in need thereof.

[00010] In yet another aspect, a method of treating or preventing uterine fibroids, dysfunctional bleeding, uterine leiomyomata, endometriosis, or polycystic ovary syndrome is provided and includes administering a deuterium enriched Tanaproget compound or a composition containing a deuterium enriched Tanaproget compound to a female in need thereof.

[00011] In still a further aspect, a method of treating or preventing benign and malignant neoplastic disease is provided and the method includes administering a deuterium enriched Tanaproget compound or a composition containing a deuterium enriched Tanaproget compound to a subject in need thereof. [00012] In another aspect, a process of preparing a deuterium enriched

Tanaproget is provided.

[00013] In yet a further aspect, a kit containing deuterium-enriched Tanaproget is provided.

[00014] In still another aspect, an internal standard is provided and contains deuterium-enriched Tanaproget.

[00015] In a further aspect, a method for measuring the amount of Tanaproget in a biological sample is provided and includes subjecting the sample to an analytical technique, wherein deuterium-enriched Tanaproget is the internal standard in the analytical technique. In one embodiment, the analytical technique is high

performance liquid chromatography-mass spectral analysis.

[00016] Other aspects and advantages of the invention will be readily apparent from the following detailed description of the invention.

DETAILED DESCRIPTION OF THE INVENTION

[00017] The present invention provides an alternate form of Tanaproget which has use in analytical, comparative, and treatment methods. Specifically, a deuterium enriched Tanaproget compound, isomer thereof, or a pharmaceutically acceptable salt thereof is provided herein. Desirably, deuterium enriched Tanaproget discussed herein performs identically to the corresponding non-deuterium-enriched Tanaproget.

[00018] The terms "deuterium-enriched" and "deuterated" as used herein are synonymous and refer to a chemical compound where one or more hydrogen atom is replaced with deuterium. In one embodiment, the deuterium-enriched Tanaproget is stable under conditions of storage and at ambient temperatures, pressures, and relative humidities. In a further embodiment, the deuterium-enriched Tanaproget described herein is about 3 to 7 mass units heavier than non-deuterium-enriched Tanaproget. In another embodiment, one or both methyl groups at the 4-position of the base molecule of Tanaproget is deuterated. In another embodiment, the deuterium enriched

Tanaproget compound is:

[00019] The deuterium-enriched Tanaproget encompasses tautomeric forms of deuterium-enriched Tanaproget and salts derived from pharmaceutically or physiologically acceptable acids, bases, alkali metals and alkaline earth metals. Also provided are derivatives of deuterium-enriched Tanaproget, such as those discussed below.

[00020] Physiologically acceptable acids include those derived from inorganic and organic acids. A number of inorganic acids are known in the art and include, without limitation, hydrochloric, hydrobromic, hydroiodic, sulfuric, nitric, and phosphoric acid. A number of organic acids are also known in the art and include, without limitation, lactic, formic, acetic, fumaric, citric, propionic, oxalic, succinic, glycolic, glucuronic, maleic, furoic, glutamic, benzoic, anthranilic, salicylic, tartaric, malonic, mallic, phenylacetic, mandelic, embonic, methanesulfonic, ethanesulfonic, panthenoic, benzenesulfonic, toluenesulfonic, stearic, sulfanilic, alginic, and galacturonic acids.

[00021] Physiologically acceptable bases include those derived from inorganic and organic bases. A number of inorganic bases are known in the art and include, without limitation, aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc sulfate or phosphate compounds, among others. Organic bases include, without limitation, Ν,Ν-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, and procaine, among others.

[00022] Physiologically acceptable alkali salts and alkaline earth metal salts include, without limitation, sodium, potassium, calcium and magnesium salts, optionally in the form of esters, and carbamates.

[00023] The deuterium-enriched Tanaproget salts can be also in the form of esters, carbamates, sulfates, ethers, oximes, carbonates, and other conventional "pro- drug" forms, which, when administered in such form, convert to the active moiety in vivo. In one embodiment, the prodrugs are esters.

[00024] The deuterium-enriched Tanaproget discussed herein also encompasses

"metabolites" which are unique products formed by processing Tanaproget by the cell or subject. In one embodiment, metabolites are formed in vivo.

[00025] A "subject" or "patient", as used herein, is a mammal. In one embodiment, the patient or subject is a human. In another embodiment, the patient or subject is a veterinary subject such as a mouse, rat, guinea pig, dog, cat, horse, cow, or pig. In a further embodiment, the patient or subject is a non-human primate such as a monkey, chimpanzee, baboon or gorilla.

[00026] As referred to herein below, the term "wt/wt" refers to the weight of one component based on the total weight of the composition. Typically, this ratio does not include the weight of any capsule utilized, the weight of any filler utilized, and seal coating, if so utilized.

[00027] The term "purified" as used herein refers to deuterium-enriched

Tanaproget that contains less than about 1% impurities. In one example, deuterium- enriched Tanaproget contains less than about 0.5% impurities. In another example, deuterium-enriched Tanaproget contains less than or equal to about 0.4% impurities. In one embodiment, deuterium-enriched Tanaproget is at least about 99% pure. In another embodiment, deuterium-enriched Tanaproget is at least about 99.5% pure. In a further embodiment, deuterium-enriched Tanaproget is at least about 99.6% pure. In yet another embodiment, deuterium-enriched Tanaproget is at least about 99.7% pure. In still a further embodiment, deuterium-enriched Tanaproget is at least about 99.8% pure. In another embodiment, deuterium-enriched Tanaproget is at least about 99.9% pure. In still a further embodiment, deuterium-enriched Tanaproget is about 100% pure.

[00028] The term "solvent" as used herein refers to a solvent in which deuterium-enriched Tanaproget has acceptable, moderate, good, or complete solubility. [00029] Deuterium-enriched Tanaproget can be milled and/or micronized. In one embodiment, deuterium-enriched Tanaproget is milled or micronized under nitrogen. Conventional milled and micronizing techniques may be utilized. In one embodiment, micronization may be performed using a Trost or jet mill, applied to non-micronized tanaproget.

[00030] Micronized and/or milled deuterium-enriched Tanaproget may have a median particle size of less than about 20 μιη, desirably less than about 15 μιη, and more desirably less than about 10 μιη. Specifically, 90% of the particles are less than or equal to about 10 μιη and 50% are less than or equal to about 3 μιη as determined by the Malvern method, which is readily understood by one of skill in the art.

[00031] Internal/Reference Standards

[00032] Further provided is a reference (or internal) standard of deuterium- enriched Tanaproget. Desirably, the reference standard is stable under conditions of manufacture and storage, and free of the contaminating action of microorganisms such as bacteria and fungi. The reference standard optionally contains a solvent in which deuterium-enriched Tanaproget is soluble and/or a solid excipient required for analysis. In one embodiment, the solvent is selected from among acetone, ethyl acetate, methyl acetate, methyl isobutyl ketone, butanol, isobutyl acetate, methanol, ethanol, 1-propanol, 2-propanol, acetonitrile, dichloromethane, among others.

Desirably, deuterium-enriched Tanaproget has acceptable, moderate, good, or complete solubility therein. In another embodiment, the deuterium-enriched

Tanaproget is combined with a solid such as potassium bromide.

[00033] The reference standard may also contain Tanaproget forms aside from deuterium-enriched Tanaproget. In one embodiment, the reference standard may contain non-deuterium enriched Tanaproget as described in US Patent No. 6,436,929. In another embodiment, the reference standard may contain polymorph Form II of Tanaproget as described in International Patent Publication No. WO-2006/1 16526. In a further embodiment, the reference standard may contain the purified Form I of Tanaproget as described in International Patent Publication No. WO-2006/1 16498. [00034] The reference standard is thereby useful for a variety of purposes. In one embodiment, the deuterium-enriched Tanaproget may be utilized for clinical work. In another embodiment, the deuterium-enriched Tanaproget may be utilized in non-clinical work.

[00035] In one embodiment, the reference/internal standard is useful for validating a sample containing Tanaproget. In this case, the reference standard contains deuterium-enriched Tanaproget. The sample being measured is obtained from the subject and contains non-deuterium-enriched Tanaproget, if any, which was not metabolized or processed by the subject. By using the reference standard, the amount of Tanaproget in the sample can be quantified.

[00036] The term sample as used herein refers to aliquots which are obtained from a subject or prepared in a laboratory lacking mammalian components. The invention described herein is not limited as to the sample being analyzed. In one embodiment, the sample is obtained from a mammalian subject and is a biomatrix such as blood, plasma, serum, urine, saliva, mucous, tissue, bile, intestinal content, feces, hair, skin, seminal fluid, tears, seat, synovial fluid, bone marrow, lymph fluid, or cerebrospinal fluid. One of skill in the art would be able to prepare such samples for analysis as described herein. Desirably, the biological properties of the sample do not affect the analysis of the deuterium-enriched Tanaproget and/or non-deuterium- enriched Tanaproget present therein.

[00037] In another embodiment, the reference/internal standard is useful for determining the purity of a sample containing Tanaproget.

[00038] In a further embodiment, the reference/internal standard is useful for standardizing a sample containing Tanaproget.

[00039] In yet another embodiment, the reference/internal standard is useful for performing quality control on a sample containing Tanaproget.

[00040] In still a further embodiment, the reference/internal standard is useful for forecasting the path of Tanaproget through the subject. [00041] In another embodiment, the reference/internal standard is useful for predicting the chemical structure of metabolites of Tanaproget formed in the body of the subject.

[00042] In yet a further embodiment, the reference/internal standard is useful for safety testing.

[00043] In still another embodiment, the reference/internal standard is useful for calibration of instruments. Desirably, the calibration is utilized in characterization of a sample containing Tanaproget, among others, using analytical techniques known to those of skill in the art.

[00044] Analytical techniques that may utilize the reference standard of deuterium-enriched Tanaproget include, without limitation, nuclear magnetic resonance (proton and carbon), mass spectroscopy (MS) optionally preceded by a separation technique, infrared spectroscopy, chromatography such as high performance liquid chromatography (HPLC), HPLC-MS, HPLC-MS/MS, HPLC- NMR, matrix-assisted laser/desorption ionization (MALDI), desorption electrospray ionization (DESI), X-ray diffraction (XRD), photodiode-array, fluorescence, polarimetry, capillary electrophoresis and differential scanning calorimetry (DSC), among others. In one embodiment, the analytical technique is a MS-liquid phase separation and detection technique. In one embodiment, the analytical technique is a MS-gas phase separation and detection technique. One of skill in the art would be able to perform such techniques.

[00045] In one embodiment, deuterium enriched Tanaproget elutes at the same time, i.e., has the same retention time, as non-deuterium enriched Tanaproget when subjected to HPLC, HPLC-MS, or HPLC-MS/MS. In another embodiment, the m/z ratio for deuterium enriched Tanaproget differs from the m/z ratio for non-deuterium enriched Tanaproget when subjected to MS alone or in combination with other spectroscopic techniques. In a further embodiment, the fragmentation pattern of deuterium-enriched Tanaproget is similar to the fragmentation pattern of non- deuterium-enriched Tanaproget when subjected to MS alone or in combination with other spectroscopic techniques. [00046] In another embodiment, deuterium-enriched Tanaproget is utilized as an internal standard for HPLC -MS/MS analysis of samples containing Tanaproget. Such samples are typically analyzed to determine the amount of Tanaproget present therein. This information is optimally utilized in therapeutic drug monitoring to optimize the therapeutic dosing, thereby being able to include different subjects' ability to absorb, metabolize, and/or excrete the administered drug. Bioequivalence studies can then be performed to compare pharmacokinetic parameters of the deuterium-enriched and non-deuterium enriched samples.

[00047] By analyzing such samples using the methods and reagents utilized therein, the absorption of Tanaproget in the subject treated with deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget can be traced, i.e., the transfer of Tanaproget from the site of administration. It is also possible to detect, measure, and analyze any metabolites present therein. Advantageously, the use of deuterium- enriched Tanaproget in such analyses is highly accurate.

[00048] The use of deuterium-enriched Tanaproget as the internal standard permits accurate quantification of the amount of non-deuterium enriched Tanaproget in the sample being analyzed, thereby preventing variability in the % extraction of non-deuterium-enriched Tanaproget from the sample. A deuterium-enriched Tanaproget internal standard serves as a means to normalize for differences in the extraction efficiencies of the non-deuterated material in different samples. By doing so, the ratio of the mass spectrometric response for the deuterium-enriched internal standard to that for the non-deuterated compound is not dependent on the extraction efficiency of the assay. In one embodiment, any effects of the biomatrix extract on the ionization of the non-deuterated analyte are also reflected in similar effects on the deuterated standard, which co-elutes with the non-deuterated analyte, thus neutralizing another potential source of assay variability.

[00049] Additional Components of Deuterium Enriched Tanaproget Compositions

[00050] Additional components may be included in the compositions discussed herein. It is advantageous that the additional components do not interfere with the function of the required components. The compositions can thereby further include other adjuvants, syrups, elixirs, diluents, excipients, binders, lubricants, surfactants, granulating agents, disintegrating agents, emollients, metal chelators, pH adjustors, fillers, antioxidants, flavoring agents, coloring agents, preservatives, and

combinations thereof, among others.

[00051] The additional components can therefore include, without limitation, one or more of vitamin E, ascorbic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), cysteine, sodium thiosulfate, povidone, cellulose, methylcellulose, hydroxymethylcellulose, carboxymethylcellulose calcium, carboxymethylcellulose sodium, hydroxypropylcellulose,

hydroxypropylmethylcellulose phthalate, noncrystalline cellulose,

polypropylpyrrolidone, polyvinylpyrrolidone (PVP), gelatin, gum arabic and acacia, polyethylene glycols, starch, sugar such as sucrose, kaolin, dextrose, and lactose, cholesterol, tragacanth, stearic acid, gelatin, casein, lecithin (phosphatide), cetostearyl alcohol, cetyl alcohol or ester wax, dextrate, dextrin, glyceryl monooleate, monostearate, or palmitostearate, polyoxyethylene alkyl ethers, castor oil derivative or stearate, polyvinyl alcohol, gelatin, light anhydrous silicic acid, talc, stearic acid, sodium lauryl sulfate (SLS), magnesium stearate, sodium stearyl furamate, silicon dioxide, starch, calcium carbonate, pectin, crospovidone (polyplasdone),

carboxymethylcellulose, sodium bicarbonate, calcium phosphate, calcium citrate, sodium starch glycolate, pregelatinized starch, crospovidone, stearyl alcohol, mink oil, cetyl alcohol, oleyl alcohol, isopropyl laurate, polyethylene glycol, olive oil, petroleum jelly, palmitic acid, oleic acid, myristyl myristate, polysorbates, sorbitan esters, poloxamer, edetic acid, malic acid, fumaric acid, citric acid, ascorbic acid, fumaric acid, or malic acid, mannitol, calcium phosphate, pregelatinized starch, or sucrose, or combinations thereof.

[00052] Treatment Methods of Using Deuterium Enriched Tanaproget

[00053] Also described are treatment methods of using deuterium enriched Tanaproget. In one embodiment, deuterium enriched Tanaproget may be utilized in methods of treating a variety of conditions affected by the progesterone receptor. In another embodiment, any condition which may be treated or prevented by using a PR agonist is contemplated for treatment with deuterium enriched Tanaproget. In a further embodiment, the deuterium-enriched Tanaproget may be utilized to analyze samples obtained from subjects treated with non-deuterium-enriched Tanaproget.

[00054] Both deuterium-enriched Tanaproget and its undeuterated form are therefore useful in a variety of conditions modulated by the progesterone receptor. In one embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful for contraception. In another embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful in hormone replacement therapy. In a further embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful in the treatment and/or prevention of uterine myometrial fibroids. In still another embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful in treating and/or preventing benign prostatic hypertrophy. In yet a further embodiment, deuterium- enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful in preventing and/or treating benign and malignant neoplastic disease. In another embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched

Tanaproget are useful in preventing and/or treating dysfunctional bleeding. In still another embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched Tanaproget are useful in treating and/or preventing uterine leiomyomata. In a further embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched

Tanaproget are useful in treating and/or preventing endometriosis. In yet another embodiment, deuterium-enriched Tanaproget and/or non-deuterium-enriched

Tanaproget are useful in treating and/or preventing polycystic ovary syndrome. In still a further embodiment, deuterium-enriched Tanaproget and/or non-deuterium- enriched Tanaproget are useful in treating and/or preventing carcinomas and adenocarcinomas. In one embodiment, the carcinoma or adenocarcinoma is of the pituitary, endometrium, kidney, ovary, breast, colon, or prostate.

[00055] Deuterium-enriched Tanaproget may be formulated into a dosing unit for delivery to a subject. Suitable dosing units include oral dosing units, such as a tablet, caplet, capsule, tablet-in-capsule, powder, suspension, microcapsule, dispersible powder, granule, suspension, syrup, elixir, and aerosol, among others. These dosing units are readily prepared using the methods described herein and those known to those of skill in the art.

[00056] Solid forms, including tablets, caplets, capsules, tablet-in-capsules, or caplet-in-capsule containing deuterium-enriched Tanaproget can be formed. The tablets or caplets that contain deuterium-enriched Tanaproget are optionally film- coated using reagents and techniques known to those skilled in the art.

[00057] A pharmaceutically effective amount of deuterium-enriched

Tanaproget can vary depending on the components of the composition, delivery mode, condition severity, the agent and weight of the subject, among others. The dosing regimen can also be adjusted to provide the optimal therapeutic response. A single dose can be delivered or several divided doses can be delivered daily, e.g., in divided doses 2 to 4 times a day. The dose can be reduced or increased as indicated by the exigencies of the therapeutic situation. In one embodiment, the delivery is on a daily, weekly, or monthly basis. In another embodiment, the delivery is on a daily delivery.

[00058] Deuterium-enriched Tanaproget may optionally be administered with one or more other progesterone receptor agonists, estrogen receptor agonists, progesterone receptor antagonists, and selective estrogen receptor modulators, chemotherapeutic agents, all of which can readily be selected by one of skill in the art.

[00059] The dosage requirements of deuterium-enriched Tanaproget or non- deuterium-enriched Tanaproget may vary based on the formulation, potency of the drug, severity of the symptoms presented, particular subject being treated including their age, weight and sex, route of delivery, and response pattern of the subject.

Treatment can be initiated with small dosages less than the optimum dose of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget. The dosage may be increased until the optimum effect under the circumstances is reached. Precise dosages will be determined by the administering physician based on experience with the individual subject treated. In one embodiment, deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is administered at a concentration that will generally afford effective results without causing any unacceptable harmful or deleterious side effects. In one embodiment, an effective amount of deuterium- enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.001 mg to 50 mg. In a further embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.01 mg to about 1 mg. In another embodiment an effective amount of deuterium-enriched Tanaproget or non- deuterium-enriched Tanaproget is 0.05 mg to about 0.3 mg. In yet another embodiment, an effective amount of deuterium-enriched Tanaproget or non- deuterium-enriched Tanaproget is about 0.01 mg. In a further embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.05 mg. In another embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.075 mg. In still a further embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.1 mg. In yet another embodiment, an effective amount of deuterium-enriched Tanaproget or non- deuterium-enriched Tanaproget is about 0.15 mg. In a further embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.2 mg. In still another embodiment, an effective amount of deuterium-enriched Tanaproget or non-deuterium-enriched Tanaproget is about 0.3 mg. In yet a further embodiment, an effective amount of deuterium-enriched Tanaproget is about 1 mg. In another embodiment, an effective amount of deuterium- enriched Tanaproget or non-deuterium-enriched Tanaproget is about 5 mg.

[00060] The deuterium-enriched Tanaproget or non-deuterium-enriched

Tanaproget may be administered to the subject by any route, taking into consideration the specific condition for which it has been selected. Such delivery may include, without limitation, topical, oral, by injection (intravascular, intravenous or intraarterial), inhalation, ocular, transdermal, subcutaneous, intramuscular, sublingual, intracranial, epidural, rectal, and vaginal, among others.

[00061] Kits Containing Deuterium Enriched Tanaproget

[00062] A kit containing a reference standard of deuterium-enriched

Tanaproget or a composition containing same is also provided. The kits containing the reference standard optionally contain a solvent, non-deuterium-enriched Tanaproget, and/or instructions for performing the kit, among others, and are described in detail below. The kit contains instructions on dosing and an insert regarding Tanaproget. Optionally, the kit may further contain instructions for monitoring local or circulating levels of product and materials for performing such assays including, e.g., reagents, well plates, containers, markers or labels, and the like. Such kits are readily packaged in a manner suitable for treatment of a desired indication. For example, the kit may also contain instructions for use of an oral dosage form such as a pill, capsule, patch, spray pump or other delivery device. Other suitable components to include in such kits will be readily apparent to one of skill in the art, taking into consideration the desired indication and the delivery route. One or more components of these kits also may be provided in dried or lyophilized forms. When reagents or components are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. It is envisioned that the solvent also may be provided in another package.

[00063] In one embodiment, the kit includes tablets, caplets, or capsules. A number of packages or kits for use in dispensing pharmaceutical agents for oral use are known in the art. In one embodiment, the kit contains tablets, caplets, or capsules which are packaged in blister packs, dial dispenser package, bottle, or desirably Ultrx™ 2000 blister packs. In another embodiment, the package/kit has indicators for each day of the 28-day cycle. When the compositions are delivered with periodic discontinuation, a package or kit can include placebos on those days when the composition is not delivered.

[00064] In one embodiment, a kit includes a single phase of a daily dosage of the composition over a 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31-day cycle. Alternatively, a kit can include a single phase of a daily dosage of the composition over the first 21 days of a 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or 31 -day cycle. A kit can also include a single phase of a daily dosage of the composition over the first 28 days of a 28, 29, 30, or 31 -day cycle.

[00065] In another embodiment, a 28-day kit can include the following, wherein the total number of the daily dosage units is 28. [00066] a first phase of from 14 to 28 daily dosage units of the composition; and a second phase of from 0 to 7 daily dosage units of an orally and

pharmaceutically acceptable placebo

[00067] a first phase of from 14 to 21 daily dosage units of the composition; a second phase of from 7 to 14 daily dosage units of an orally and pharmaceutically acceptable placebo

[00068] a first phase of from 18 to 21 daily dosage units of the composition; a second phase of from 7 to 10 daily dose units of a pharmaceutically acceptable placebo

[00069] a first phase of 14 daily dosage units of the composition; and a second phase of 14 daily units of an orally and pharmaceutically acceptable placebo

[00070] a first phase of 17 daily dosage units of the composition; and a second phase of 1 1 daily units of an orally and pharmaceutically acceptable placebo

[00071] a first phase of 21 daily dosage units of the composition; and a third phase of 7 daily units of an orally and pharmaceutically acceptable placebo.

[0001] In one embodiment, the daily dosage of deuterium-enriched Tanaproget remains fixed in each phase. In another embodiment, the daily dosage of deuterium- enriched Tanaproget varies in each phase. The daily dose units described may be delivered in the order described, with the first phase followed in order by the second and third phases, etc., or may be varied.

[0002] The kit can further contain instructions for administering the deuterium- enriched Tanaproget composition.

[00072] Processes for Preparing Compounds of the Invention

[00073] Also provided herein are processes for preparing compounds of the invention. In one embodiment, process for preparing deuterium enriched Tanaproget are provided. In another embodiment, processes for producing deuterium enriched Tanaproget (wherein the methyl groups at the 4-position are deuterated) are provided. In a further embodiment, processes for producing the following compound are provided:

[00074] Methods of preparing non-deuterated Tanaproget are described in US

Patent Nos. 6,436,929 and 7,786,297 and US Patent Application Publication No. 2005/0272702, which are herein incorporated by reference. In one embodiment, deuterium-enriched Tanaproget may be prepared as described in Scheme 1.

[00076] Scheme 1 sets forth the preparation of deuterium enriched Tanaproget.

In this scheme, methyl 2-amino-5-bromobenzoate (I) is reacted with at least 2 equivalents of a deuterated methylating agent to provide compound II. In one embodiment, about 2 to about 6 equivalents of the methylating agent may be utilized. In a further embodiment, the methylating agent is CDsMgl, CDsMgCl, CDsMgBr, or CD₃L1. In another embodiment, the CDsMgl is added to the methyl 2-amino-5- bromobenzoate solution. In still another embodiment, the methyl 2-amino-5- bromobenzoate solution is added to the CDsMgl. In a further embodiment, the methylation of compound I is performed at reduced temperatures. In yet another embodiment, the methylation of compound I is performed at about -78°C to about - 50°C. The methylation is performed in a solvent such as an ether such as

tetrahydrofuran, diethylether, dimethoxyethane, dioxane, dioxolane,

methylcyclopentyl ether. Finally, compound II may be isolated using techniques known to those skilled in the art.

[00077] Compound II is then reacted with triphosgene, 1, 1'- carbonyldiimidazole, dimethylcarbonate or diethylcarbonate to provide 6-bromo dimethyl dihydrobenzoxazinone (III). The reaction may be performed in an inert solvent such as an aprotic solvent. In one embodiment, the aprotic solvent is an ether. In another embodiment, the inert solvent is tetrahydrofuran, toluene, or cyclopentyl methyl ether (CPME). In a further embodiment, this reaction is performed at reduced temperatures. In another embodiment, this reaction is performed at a temperature of about 0°C to about 10°C. In a further embodiment, the triphosgene or 1,1'- carbonyldiimidazole is added to compound II. Finally, compound III may be isolated using techniques known to those skilled in the art. In one embodiment, compound III may be recrystallized using techniques, skill, and reagents known in the art. In another embodiment, compound III is recrystallized using acetonitrile.

[00078] Compound III is then coupled with a 1 -methyl-2-cyanopyrrole moiety to provide compound VII. In one embodiment, compound III is reacted with an aryl boronic acid, aryl stannane or aryl zinc compound bearing a 5 -cyano-1 -methyl- 1H- pyrrol-2-yl substituent. In one embodiment, compound III is reacted with borate- 1- (5-cyano-l-methyl-lH-pyrrol-2-yl)tris(2-propanolato)lithium (V). In one

embodiment, borate- 1 -(5-cyano- 1 -methyl- lH-pyrrol-2-yl)tris(2-propanolato)lithium is added to the 6-bromo dimethyl dihydrobenzoxazinone. In another embodiment, this coupling reaction is performed in the presence of a palladium catalyst.

[00079] In a further embodiment, the reaction is performed in the presence of a palladium or nickel catalyst. In one embodiment, the catalyst is a palladium or nickel complex with phosphino ligands such as Ph₃P, dppf (l, l'-bis-diphenylphosphino ferrocene), or dppe (l,2-bis(diphenylphosphino)ethane). In another embodiment the catalyst is palladium tetrakis(triphenylphosphine). In a further embodiment, the catalyst is palladium acetate. The reaction may be performed in an inert solvent such as tetrahydrofuran, benzene, DMF, isopropanol, ethanol, DME, ether, acetone or a mixture of above solvents, optionally in the presence of water. In yet another embodiment, the coupling reaction may be performed elevated temperatures. Finally, compound VII may be isolated using techniques known to those skilled in the art. Compound VII may optionally be recrystallized using techniques, skill, and reagents known in the art. In one embodiment, compound VII is recrystallized using toluene or acetone/water.

[00080] Finally, compound VII is reacted with a thionating agent to provide compound VIII. In one embodiment, the thionating agent is Lawesson's reagent, P2S5, or bistrimethylsylilsulfide. The reaction may be performed in an inert solvent such as toluene or dimethoxyethane. In one embodiment, the thionation is performed at elevated temperatures. Finally, compound VIII may be isolated using techniques known to those skilled in the art.

[00081] Compound VIII may be purified using techniques, skill, and reagents known in the art. In one embodiment, compound VIII is purified using

recrystallization, chromatography, or combinations thereof. In a further embodiment, compound VIII is purified using flash chromatography. In another embodiment, deuterium enriched Tanaproget is recrystallized from acetone/water, methyl ethyl ketone, ethylene glycol, dimethylformamide (DMF)/water, n-heptane, tetrahydrofuran (THF)/water, pentane, dimethylacetamide (DMA)/water, methyl cyclohexane, sec- butanol, 1,2-dichlroethane, tert-butyl methyl ether, 2-methyl-l-propanol, butyl acetate, isobutyl acetate, cyclopentanone, methyl isobutyl ketone, diethyl ether, methyl THF, l-methyl-2-pyrrolidinone, n-butanol, isopropyl acetate, methanol, ethanol, 1-propanol, 2-propanol, acetone, acetonitrile, ethylacetate, dichloromethane. In a further embodiment, recrystallization of deuterium enriched Tanaproget includes dissolving deuterium enriched Tanaproget in acetone as step a. Step b then includes warming the solution of step (a). In one embodiment, step b is performed at a temperature of about 50°C to about reflux temperatures of the solvent. Step c includes adding water optionally in the presence of an anti-solvent such as heptane, hexane, diethylether, MTBE, without limitation, to the solution of step (b). In one embodiment, the ratio of acetone:water is at least about 2: 1. In another embodiment, the ratio of acetone:water is about 3 : 1. Step d include cooling the solution of step (c). In one embodiment, the solution is cooled to about -15°C to about 10°C. As step e, the solid from step (d) is isolated using techniques and skill in the art. Finally, the isolated product is dried using techniques and skill in the art.

[00082] The following examples are illustrative only and are not intended to limit the present invention.

[00083] EXAMPLES

[00084] Example 1: Preparation of d⁶-6-bromo dimethyl

dihydrobenzoxazinone (III)

[00085] To a suspension of magnesium (4.2 g; 175 mmol) in diethyl ether (100 mL) at 25°C was added about 1/10 of a solution of iodomethane-d³ (25 g 170 mmol) in diethyl ether (75 mL). After 15 minutes, the temperature was slowly increased to the reflux temperature. The remaining d³-iodomethane solution was added dropwise, keeping the suspension to gently reflux. At the end of addition, the solution (with a little amount of unreacted magnesium) was added to a solution of methyl 2-amino-5- bromobenzoate (I, 6.5 g; 28 mmol) in THF (65 mL) and the solution cooled at -60°C. The mixture as a yellow suspension was warmed to 20°C and stirred at this temperature overnight. The mixture was then added to a water/ice mixture (100 g) and the pH of the two phase system was adjusted to 7.2 by addition of 18% hydrochloric acid. After phase separation, the organic phase was evaporated to dryness under vacuum. The residue was taken up with tetrahydrofuran (THF; 70 mL) and the insoluble salt filtered off and discharged. The THF solution was evaporated under vacuum to provide a residue (II, 5.6 g) that was dissolved in THF (60 mL). The solution was cooled to 6°C and triphosgene (7.4 g) was added portion-wise keeping the temperature below 10°C. The temperature was increased to 20°C and, after 3 hours, the solvent was evaporated under vacuum to give a residue. To the residue, methylene chloride (100 mL) and water (100 mL) were added and the pH was adjusted to 6-7 by addition of 15% sodium hydroxide. After phase separation, the organic phase was evaporated under vacuum to a residue. Acetonitrile (25 mL) was added to the residue and the resulting suspension was cooled to 0-5°C. The product was filtered off and dried at 40°C under vacuum for 4 hour to provide the product III (3.5 g HPLC purity 99.4%).

[00086] Example 2: Preparation of d⁶-5-(4,4-dimethyl-2-oxo-l,4-dihydro-

2H-3,l-benzoxazin-6-yl)-l-methyl-lH-pyrrole-2-carbonitrile (VII)

[00087] To a solution of N-methyl 2-pyrrolcarbonitrile (IV, 3.1 g; 29 mmol) in

THF (22 mL), tri-isopropyl borate (5.5 g 29 mmol) was added. The solution was cooled to 6-8°C and 2M lithium diisopropylamide (LDA; 19 mL; 38 mmol) was added dropwise over about 1 hour. The suspension was stirred for 1 hour and methanol (1.5 g 47 mmol) was added.

[00088] The resulting solution (V) was transferred at 20°C to a dropping funnel and added over about 3 hours to a well-stirred mixture of d⁶-6-bromo dimethyl dihydrobenzoxazinone (VI, 4.0 g 15 mmol), THF (32 mL), water (13.8 mL), potassium carbonate (4.2 g 30 mmol) and palladium tetrakis(triphenylphosphine) (0.29 g 0.25 mmol) kept at reflux (about 67°C). The reaction mixture was kept at reflux for 1 hour, cooled at 25°C, and water (25 mL) was the added. The pH was adjusted to 5-6 by addition of 18% hydrochloric acid and the phases were separated. The organic phase was evaporated under vacuum to a residue. Toluene (30 mL) was added to the residue and the resulting suspension was cooled to 0-5°C. The solid was filtered off and dried at 50°C under vacuum for 4 hours to provide the product VII (4.8 g; HPLC purity 90.7%).

[00089] Example 3: Preparation of d⁶-5-(4,4-dimethyl-2-thioxo-l,4- dihydro-2H-3,l-benzoxazin-6-yl)-l-methyl-lH-pyrrole-2-carbonitrile (VII)

[00090] A mixture of d⁶-5-(4,4-dimethyl-2-oxo-l,4-dihydro-2H-3,l- benzoxazin-6-yl)- 1 -methyl- lH-pyrrole-2-carbonitrile (VII, 4.8 g), toluene (32 mL), acetonitrile (3.2 mL) and Lawesson's reagent (3.6 g; 9 mmol) was heated at 90°C for 12 hours. The solvents were evaporated and the residue (VIII) was purified by flash chromatography on silica gel (mobile phase 7:3 hexane/ethyl acetate). The crude product (2.3 g) was dissolved at 56°C in acetone (16 mL). Water (8 mL) was added and the suspension was cooled to 10°C for about 2 hours. The solid was filtered off and dried at 25°C under vacuum for 3 hours to provide the purified product (1.6 g; HPLC purity 99.7%).

[00091] All publications and priority applications, including US Provisional Patent Application No. 61/782,587, filed March 14, 2013, cited in this specification are incorporated herein by reference. While the invention has been described with reference to particular embodiments, it will be appreciated that modifications can be made without departing from the spirit of the invention. Such modifications are intended to fall within the scope of the appended claims.

Claims

WHAT IS CLAIMED IS:

1. A deuterium enriched Tanaproget compound, isomer thereof, or a

pharmaceutically acceptable salt thereof.

2. The deuterium enriched Tanaproget compound according to claim 1, wherein the methyl groups at the 4-position are deuterated.

3. The deuterium enriched Tanaproget compound according to claim 1, which is:

4. A reference standard comprising the deuterium enriched Tanaproget compound of any one of claims 1 to 3.

5. The reference standard according to claim 4, further comprising a solvent.

6. The reference standard according to claim 4, further comprising non- deuterium enriched Tanaproget.

7. A composition comprising the deuterium-enriched Tanaproget compound of any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

8. The composition according to claim 7, comprising about 0.1 to about 5 mg of said deuterium enriched Tanaproget compound.

9. The composition according to claim 7, which is in tablet, caplet, capsule, or tablet-in-capsule form.

10. A method of contraception, said method comprising administering said deuterium enriched Tanaproget compound of any one of claims 1 to 3 to a female in need thereof.

11. A method of contraception, said method comprising administering a composition of any one of claims 7 to 9 to a female in need thereof.

12. A method of hormone replacement therapy, said method comprising administering said deuterium enriched Tanaproget compound of any one of claims 1 to 3 to a subject in need thereof.

13. A method of hormone replacement therapy, said method comprising administering a composition of any one of claims 7 to 9 to a subject in need thereof.

14. A method of treating or preventing uterine fibroids, dysfunctional bleeding, uterine leiomyomata, endometriosis, or polycystic ovary syndrome, said method comprising administering a compound of any one of claims 1 to 3 to a female in need thereof.

15. A method of treating or preventing uterine fibroids, dysfunctional bleeding, uterine leiomyomata, endometriosis, or polycystic ovary syndrome, said method comprising administering a composition of any one of claims 7 to 9 to a female in need thereof.

16. A method of treating or preventing benign and malignant neoplastic disease, said method comprising administering said deuterium enriched Tanaproget compound of any one of claims 1 to 3 to a subject in need thereof.

17. A method of treating or preventing benign and malignant neoplastic disease, said method comprising administering a composition of any one of claims 7 to 9 to a subject in need thereof.

18. A process of preparing deuterium enriched Tanaproget, said process comprising:

(i) reacting methyl 2-amino-5-bromobenzoate with at least 2 equivalents of d³-iodomethane;

(ii) reacting the product of step (i) with triphosgene or 1 , Γ- carbonyldiimidazole to provide 6-bromo dimethyl dihydrobenzoxazinone;

(iii) reacting 6-bromo dimethyl dihydrobenzoxazinone with Borate- 1 -(5- cyano- 1 -methyl- 1 H-pyrrol-2-yl)tris(2-propanolato)lithium; and

(iv) reacting the product of step (iii) with Lawesson's reagent.

19. The process according to claim 18, wherein said d3-iodomethane is added to said methyl 2-amino-5-bromobenzoate.

20. The process according to claim 18, wherein step (i) is performed at reduced temperatures.

21. The process according to claim 18, wherein said reduced temperature is about -78°C to about -50°C.

22. The process according to any one of claims 18 to 21, wherein step (i) is performed in tetrahydrofuran, diethylether, dimethoxyethane, or dioxane.

23. The process according to any one of claims 18 to 21, wherein the product of step (i) is isolated.

24. The process according to any one of claims 18 to 21, wherein step (ii) is performed in tetrahydrofuran, toluene, or CP ME.

25. The process according to any one of claims 18 to 21, wherein step (ii) is performed at reduced temperatures.

26. The process according to claim 25, wherein said reduced temperature is about 0°C to about 10°C.

27. The process according to any one of claims 18 to 21, wherein said triphosgene is added to the product of step (i).

28. The process according to any one of claims 18 to 21, wherein the product of step (ii) is isolated.

29. The process according to claim 28, wherein the product of step (ii) is recrystallized from acetonitrile.

30. The process according to any one of claims 18 to 21, wherein step (iii) is performed in the presence of a palladium catalyst.

31. The process according to claim 30, wherein said catalyst is palladium tetrakis(triphenylphosphine).

32. The process according to any one of claims 18 to 21, wherein step (iii) is performed in tetrahydrofuran.

33. The process according to any one of claims 18 to 21, wherein said cyanopyrrole is added to said 6-bromo dimethyl dihydrobenzoxazinone.

34. The process according to any one of claims 18 to 21, wherein step (iii) is performed at elevated temperatures.

35. The process according to any one of claims 18 to 21, wherein said deuterium enriched Tanaproget is isolated.

36. The process according to any one of claims 18 to 21, wherein said deuterium enriched Tanaproget is recrystallized from toluene.

37. The process according to any one of claims 18 to 21, wherein step (iv) is performed in toluene.

38. The process according to any one of claims 18 to 21, wherein step (iv) is performed at elevated temperatures.

39. The process according to any one of claims 18 to 21, further comprising (v) purifying the product of step (iv).

40. The process according to claim 39, wherein said purification comprises flash chromatography.

41. The process according to claim 40, wherein said deuterium enriched Tanaproget is recrystallized from acetone/water.

42. The process according to claim 41, wherein said recrystallization comprises:

(a) dissolving said deuterium enriched Tanaproget in acetone;

(b) warming the solution of step (a);

(c) adding water to the solution of step (b), wherein the ratio of acetone:water is 2: 1 ;

(d) cooling the solution of step (c) to about -15°C to about 10°C;

(e) isolating the solid from step (d); and

(f) drying said solid from step (e).

43. The process according to claims 42, which yields 99.7% pure deuterium enriched Tanaproget.

44. A kit comprising said deuterium-enriched Tanaproget of any one of claims 1 to 3.

45. An internal standard comprising said deuterium-enriched Tanaproget of any one of claims 1 to 3.

46. A method for measuring the amount of Tanaproget in a biological sample, said method comprising subjecting said sample to an analytical technique, wherein said deuterium-enriched Tanaproget of any one of claims 1 to 3 is the internal standard in said analytical technique.

47. The method according to claim 46, wherein said analytical technique is chromatography such as high performance liquid chromatography (HPLC), or HPLC- MS, HPLC-MS/MS.

48. Use of said deuterium enriched Tanaproget compound of any one of claims 1 to 3 in the preparation of medicament for contraception in a female in need thereof.

49. Use of said deuterium enriched Tanaproget compound of any one of claims 1 to 3 in the preparation of a medicament for hormone replacement therapy in a subject in need thereof.

50. Use of said deuterium enriched Tanaproget compound of any one of claims 1 to 3 in the preparation of a medicament for treating or preventing uterine fibroids, dysfunctional bleeding, uterine leiomyomata, endometriosis, or polycystic ovary syndrome in a female in need thereof.

51. Use of said deuterium enriched Tanaproget compound of any one of claims 1 to 3 in the preparation of a medicament for treating or preventing benign and malignant neoplastic disease in a subject in need thereof.