Treatment of hair loss disorders with deuterated jak inhibitors

RELATED APPLICATIONS

0001This application claims the benefit of <patcit id="pcit0001" dnum="US62331827"><text>U.S. Provisional Application Nos. 62/331,827, filed on May 4, 2016</text></patcit>, <patcit id="pcit0002" dnum="US62338869"><text>62/338,869, filed on May 19, 2016</text></patcit>, <patcit id="pcit0003" dnum="US62418774"><text>62/418,774, filed on November 7, 2016</text></patcit>, <patcit id="pcit0004" dnum="US62419237"><text>62/419,237, filed on November 8, 2016</text></patcit>, <patcit id="pcit0005" dnum="US62434404"><text>62/434,404, filed on December 14, 2016</text></patcit>, <patcit id="pcit0006" dnum="US62466358"><text>62/466,358, filed on March 2, 2017</text></patcit> and <patcit id="pcit0007" dnum="US62492758"><text>62/492,758, filed on May 1, 2017</text></patcit>. The entire teachings of the above applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

0002Many current medicines suffer from poor absorption, distribution, metabolism and/or excretion (ADME) properties that prevent their wider use or limit their use in certain indications. Poor ADME properties are also a major reason for the failure of drug candidates in clinical trials. While formulation technologies and prodrug strategies can be employed in some cases to improve certain ADME properties, these approaches often fail to address the underlying ADME problems that exist for many drugs and drug candidates. One such problem is rapid metabolism that causes a number of drugs, which otherwise would be highly effective in treating a disease, to be cleared too rapidly from the body. A possible solution to rapid drug clearance is frequent or high dosing to attain a sufficiently high plasma level of drug. This, however, introduces a number of potential treatment problems such as poor patient compliance with the dosing regimen, side effects that become more acute with higher doses, and increased cost of treatment. A rapidly metabolized drug may also expose patients to undesirable toxic or reactive metabolites.

0003Another ADME limitation that affects many medicines is the formation of toxic or biologically reactive metabolites. As a result, some patients receiving the drug may experience toxicities, or the safe dosing of such drugs may be limited such that patients receive a suboptimal amount of the active agent. In certain cases, modifying dosing intervals or formulation approaches can help to reduce clinical adverse effects, but often the formation of such undesirable metabolites is intrinsic to the metabolism of the compound.

0004In some select cases, a metabolic inhibitor will be co-administered with a drug that is cleared too rapidly. Such is the case with the protease inhibitor class of drugs that are used to treat HIV infection. The FDA recommends that these drugs be co-dosed with ritonavir, an inhibitor of cytochrome P450 enzyme 3A4 (CYP3A4), the enzyme typically responsible for their metabolism (see <nplcit id="ncit0001" npl-type="s"><text>Kempf, D.J. et al., Antimicrobial agents and chemotherapy, 1997, 41(3): 654-60</text></nplcit>). Ritonavir, however, causes adverse effects and adds to the pill burden for HIV patients who must already take a combination of different drugs. Similarly, the CYP2D6 inhibitor quinidine has been added to dextromethorphan for the purpose of reducing rapid CYP2D6 metabolism of dextromethorphan in a treatment of pseudobulbar affect. Quinidine, however, has unwanted side effects that greatly limit its use in potential combination therapy (see <nplcit id="ncit0002" npl-type="s"><text>Wang, L et al., Clinical Pharmacology and Therapeutics, 1994, 56(6 Pt 1): 659-67</text></nplcit>; and FDA label for quinidine at www.accessdata.fda.gov).

0005In general, combining drugs with cytochrome P450 (CYP) inhibitors is not a satisfactory strategy for decreasing drug clearance. The inhibition of a CYP enzyme's activity can affect the metabolism and clearance of other drugs metabolized by that same enzyme. CYP inhibition can cause other drugs to accumulate in the body to toxic levels.

0006A potentially attractive strategy for improving a drug's metabolic properties is deuterium modification. In this approach, one attempts to slow the CYP-mediated metabolism of a drug or to reduce the formation of undesirable metabolites by replacing one or more hydrogen atoms with deuterium atoms. Deuterium is a safe, stable, non-radioactive isotope of hydrogen. Compared to hydrogen, deuterium forms stronger bonds with carbon. In select cases, the increased bond strength imparted by deuterium can positively impact the ADME properties of a drug, creating the potential for improved drug efficacy, safety, and/or tolerability. At the same time, because the size and shape of deuterium are essentially identical to those of hydrogen, replacement of hydrogen by deuterium would not be expected to affect the biochemical potency and selectivity of the drug as compared to the original chemical entity that contains only hydrogen.

0007Over the past 35 years, the effects of deuterium substitution on the rate of metabolism have been reported for a very small percentage of approved drugs (see, e.g., <nplcit id="ncit0003" npl-type="s"><text>Blake, MI et al, J Pharm Sci, 1975, 64:367-91</text></nplcit>; <nplcit id="ncit0004" npl-type="s"><text>Foster, AB, Adv Drug Res, 1985, 14:1-40</text></nplcit> ("Foster"); <nplcit id="ncit0005" npl-type="s"><text>Kushner, DJ et al, Can J Physiol Pharmacol, 1999, 79-88</text></nplcit>; <nplcit id="ncit0006" npl-type="s"><text>Fisher, MB et al, Curr Opin Drug Discov Devel, 2006, 9:101-09</text></nplcit> ("Fisher")). The results have been variable and unpredictable. For some compounds deuteration caused decreased metabolic clearance <i>in vivo.</i> For others, there was no change in metabolism. Still others demonstrated increased metabolic clearance. The variability in deuterium effects has also led experts to question or dismiss deuterium modification as a viable drug design strategy for inhibiting adverse metabolism (see Foster at p. 35 and Fisher at p. 101).

0008The effects of deuterium modification on a drug's metabolic properties are not predictable even when deuterium atoms are incorporated at known sites of metabolism. Only by actually preparing and testing a deuterated drug can one determine if and how the rate of metabolism will differ from that of its non-deuterated counterpart. <i>See,</i> for example, <nplcit id="ncit0007" npl-type="s"><text>Fukuto et al. (J. Med. Chem., 1991, 34, 2871-76</text></nplcit>). Many drugs have multiple sites where metabolism is possible. The site(s) where deuterium substitution is required and the extent of deuteration necessary to see an effect on metabolism, if any, will be different for each drug.

0009Ruxolitinib phosphate, is a heteroaryl-substituted pyrrolo[2,3-d]pyrimidines also known as 3(R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate and as (R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile phosphate, inhibits Janus Associated Kinases (JAKs) JAK1 and JAK2. These kinases mediate the signaling of a number of cytokines and growth factors important for hematopoiesis and immune function. JAK signaling involves recruitment of STATs (signal transducers and activators of transcription) to cytokine receptors, activation and subsequent localization of STATs to the nucleus leading to modulation of gene expression.

0010Ruxolitinib phosphate is currently approved for the treatment of patients with intermediate or high-risk myelofibrosis, including primary myelofibrosis, post-polycythemia vera myelofibrosis and post-essential thrombocythemia myelofibrosis. Ruxolitinib phosphate is also currently in clinical trials for the treatment of additional conditions.

0011Despite the beneficial activities of ruxolitinib, there is a continuing need for new compounds to treat the aforementioned diseases and conditions.

SUMMARY OF THE INVENTION

0012It has now been found that deuterated analogs of ruxolitinib (including Compound (I), also referred to as (R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyclopentyl-2,2,3,3,4,4,5,5-ds)propanenitrile, or D8-ruxolitinib), are useful for the treatment of hair-loss disorders, including alopecia areata. Compound (I) is represented by the following structural formula: <chemistry id="chem0001" num="0001"><img file="EP4424367A2_D0001.tif" /></chemistry>

0013In certain embodiments, Compound (I) is administered as a pharmaceutically acceptable salt, such as the phosphate salt. Compound (I) can be administered in doses in the range of about 4 mg to about 50 mg per day (or the equivalent weight based on a salt, such as Compound (I) phosphate salt), administered as a single daily dose or in divided doses (e.g., twice per day). Based on these discoveries, novel therapies using Compound (I) or a pharmaceutically acceptable salt thereof, for treating a hair loss disorder in a mammalian subject are disclosed herein.

0014One aspect of the invention is a method for treating hair loss disorders that can be treated by compounds that modulate the activity of Janus Associated Kinase 1 (JAK1) and/or Janus Associated Kinase 2 (JAK2). The method comprises administering to a subject (e.g., a mammalian subject) an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt)), once or twice per day, wherein the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is in the range of about 4 mg/day to about 50 mg/day, for example, about 5 mg/day, about 10 mg/day, about 20 mg/day, about 30 mg/day, about 40 mg/day, or about 50 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg/day, 8 mg/day, 16 mg/day, 32 mg/day or 48 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof is 4 mg, 8mg, 12 mg or 16 mg twice per day. In certain embodiments, the hair loss disorder is alopecia areata. In certain embodiments, the subject is a human. Preferably, Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), is administered orally at any of the foregoing dosages. Preferably, the Compound (I), or a pharmaceutically acceptable salt thereof, is administered orally at any of the foregoing dosages in a pharmaceutical formulation which is a tablet.

0015In an alternative aspect, the invention provides a method for treating hair loss disorders, the method comprising topically administering to a subject (e.g., a mammalian subject) an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt). In certain embodiments, the compound is administered in a pharmaceutical composition which is formulated for topical administration, such as a cream, ointment, lotion, foam or the like.

0016In another aspect, the invention provides a method for inducing hair growth in a subject. The method comprises administering to a mammalian subject an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), once or twice per day, wherein the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is in the range of about 4 mg/day to about 50 mg/day, for example, about 5 mg/day, about 10 mg/day, about 20 mg/day, about 30 mg/day, about 40 mg/day, or about 50 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg/day, 8 mg/day, 16 mg/day, 32 mg/day or 48 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day. In certain embodiments, the subject suffers from a hair loss disorder; in further embodiments, the hair loss disorder is alopecia areata. In certain embodiments, the subject is a human. In one embodiment, the subject is a human 6 years of age or older. Preferably, Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), is administered orally at any of the foregoing dosages. Preferably, the Compound (I), or a pharmaceutically acceptable salt thereof, is administered orally at any of the foregoing dosages in a pharmaceutical formulation which is a tablet.

0017Another aspect of the invention is a method for treating autoimmune skin disorders disorders that can be treated by compounds that modulate the activity of Janus Associated Kinase 1 (JAK1) and/or Janus Associated Kinase 2 (JAK2). The method comprises administering to a subject (e.g., a mammalian subject) an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), once or twice per day, wherein the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is in the range of about 4 mg/day to about 50 mg/day, for example, about 5 mg/day, about 10 mg/day, about 20 mg/day, about 30 mg/day, about 40 mg/day, or about 50 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg/day, 8 mg/day, 16 mg/day, 32 mg/day or 48 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof is 4 mg, 8mg, 12 mg or 16 mg twice per day. In certain embodiments, the autoimmune skin disorder is alopecia areata, vitiligo, atopic dermatitis (ezcema), or psoriasis. In certain embodiments, the subject is a human. Preferably, Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), is administered orally at any of the foregoing dosages. Preferably, the Compound (I), or a pharmaceutically acceptable salt thereof, is administered orally at any of the foregoing dosages in a pharmaceutical formulation which is a tablet.

0018Another aspect of the invention is Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), for use in treating hair loss disorders that can be treated by compounds that modulate the activity of Janus kinase 1 (JAK1) and/or Janus kinase 2 (JAK2). The compound may be administered at the dosing regimens disclosed herein. In certain embodiments, the hair loss disorder is alopecia areata.

0019Still another aspect of the invention is the use of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), for the manufacture of a medicament for method for treating hair loss disorders that can be treated by compounds that modulate the activity of Janus Associated Kinase 1 (JAK1) and/or Janus Associated Kinase 2 (JAK2). The compound may be administered at the dosing regimens disclosed herein. In certain embodiments, the hair loss disorder is alopecia areata.

0020Another aspect of the invention is a pharmaceutical composition comprising Compound (I), in the range of about 4 mg to about 50 mg (for example, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg), or an equivalent amount of a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), together with a pharmaceutically acceptable carrier or diluent. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg, 8 mg, 16 mg, 24 mg, 32 mg or 48 mg. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 4 mg, 8 mg, 12 mg, or 16 mg. In certain embodiments, the pharmaceutical composition is a tablet.

BRIEF DESCRIPTION OF THE DRAWINGS

0021<ul id="ul0001" list-style="none" compact="compact"><li><figref idref="f0001 f0002">FIG. 1</figref> shows the design and results from a single ascending dose (SAD) trial in healthy volunteers. <figref idref="f0001">FIG. 1A</figref> depicts the SAD study design; <figref idref="f0001">FIG. 1B</figref> is a graph showing plasma concentration of CTP-543 (Compound (I)) from 0 - 48 hours after dosing; <figref idref="f0002">FIG.1C</figref> is a table showing mean PK parameters for CTP-543 (Compound (I)) in the SAD study.</li><li><figref idref="f0003 f0004 f0005">FIG. 2</figref> shows the design and results from a multiple ascending dose (MAD) trial in healthy volunteers. <figref idref="f0003 f0004 f0005">FIG. 2</figref><figref idref="f0003">2A</figref> depicts the MAD study design; <figref idref="f0004">FIG. 2B</figref> is a graph showing plasma concentration of CTP-543 (Compound (I)) from 0 - 24 hours on Day 1 and Day 7 of the MAD study; <figref idref="f0005">FIG. 2C</figref> is a table showing mean PK parameters for CTP-543 (Compound (I)) in the MAD study.</li></ul>

DETAILED DESCRIPTION OF THE INVENTION

<u>Definitions</u>

0022The term "treat" means decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease (e.g., a disease or disorder delineated herein), lessen the severity of the disease or improve the symptoms associated with the disease. For example, treatment of a hair loss disorder includes regrowth of hair, prevention of further hair loss, or diminishing the rate of hair loss.

0023"Hair loss disorder" means any condition or disorder that results in loss of hair on one or more areas of the body. Hair loss disorders include, without limitation, androgenetic alopecia, alopecia areata, telogen effluvium, alopecia areata, alopecia totalis, and alopecia universalis.

0024The term "mammal", as used herein, includes humans, as well as non-human mammals such as cats, dogs, sheep, cattle, pigs, goats, non-human primates (including monkeys and apes) and the like.

0025It will be recognized that some variation of natural isotopic abundance occurs in a synthesized compound depending upon the origin of chemical materials used in the synthesis. Thus, a preparation of ruxolitinib will inherently contain small amounts of deuterated isotopologues. The concentration of naturally abundant stable hydrogen and carbon isotopes, notwithstanding this variation, is small and immaterial as compared to the degree of stable isotopic substitution of compounds of this invention. See, for instance, <nplcit id="ncit0008" npl-type="s"><text>Wada, E et al., Seikagaku, 1994, 66:15</text></nplcit>; <nplcit id="ncit0009" npl-type="s"><text>Gannes, LZ et al., Comp Biochem Physiol Mol Integr Physiol, 1998, 119:725</text></nplcit>.

0026In Compound (I), any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as "H" or "hydrogen", the position is understood to have hydrogen at its natural abundance isotopic composition. Also unless otherwise stated, when a position is designated specifically as "D" or "deuterium", the position is understood to have deuterium at an abundance that is at least 3000 times greater than the natural abundance of deuterium, which is 0.015% (i.e., at least 45% incorporation of deuterium).

0027The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance and the natural abundance of a specified isotope.

0028In other embodiments, a compound of this invention has an isotopic enrichment factor for each designated deuterium atom of at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

0029The term "isotopologue" refers to a species in which the chemical structure differs from Compound (I) only in the isotopic composition thereof.

0030The term "compound," when referring to a compound of this invention, refers to a collection of molecules having an identical chemical structure, except that there may be isotopic variation among the constituent atoms of the molecules. Thus, it will be clear to those of skill in the art that a compound represented by a particular chemical structure containing indicated deuterium atoms, will also contain lesser amounts of isotopologues having hydrogen atoms at one or more of the designated deuterium positions in that structure. The relative amount of such isotopologues in a compound of this invention will depend upon a number of factors including the isotopic purity of deuterated reagents used to make the compound and the efficiency of incorporation of deuterium in the various synthesis steps used to prepare the compound.

0031The invention also provides salts of Compound (I). A salt of a compound of this invention is formed between an acid and a basic group of the compound, such as an amino functional group, or a base and an acidic group of the compound, such as a carboxyl functional group. According to another embodiment, the compound is a pharmaceutically acceptable acid addition salt, such as a phosphate salt.

0032The term "pharmaceutically acceptable," as used herein, refers to a component that is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other mammals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. A "pharmaceutically acceptable salt" means any non-toxic salt that, upon administration to a recipient, is capable of providing, either directly or indirectly, a compound of this invention. A "pharmaceutically acceptable counterion" is an ionic portion of a salt that is not toxic when released from the salt upon administration to a recipient.

0033Acids commonly employed to form pharmaceutically acceptable salts include inorganic acids such as hydrogen bisulfide, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and phosphoric acid, as well as organic acids such as para-toluenesulfonic acid, salicylic acid, tartaric acid, bitartaric acid, ascorbic acid, maleic acid, besylic acid, fumaric acid, gluconic acid, glucuronic acid, formic acid, glutamic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, lactic acid, oxalic acid, para-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid and acetic acid, as well as related inorganic and organic acids. Such pharmaceutically acceptable salts thus include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-l,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2- sulfonate, mandelate and other salts. In one embodiment, pharmaceutically acceptable acid addition salts include those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and especially those formed with organic acids such as maleic acid.

0034The term "stable compounds," as used herein, refers to compounds which possess stability sufficient to allow for their manufacture and which maintain the integrity of the compound for a sufficient period of time to be useful for the purposes detailed herein (e.g., formulation into therapeutic products, intermediates for use in production of therapeutic compounds, isolatable or storable intermediate compounds, treating a disease or condition responsive to therapeutic agents).

0035"D" and "d" both refer to deuterium. "Stereoisomer" refers to both enantiomers and diastereomers. "Tert" and "t-" each refer to tertiary. "US" refers to the United States of America.

0036"Substituted with deuterium" refers to the replacement of one or more hydrogen atoms with a corresponding number of deuterium atoms.

0037In one aspect, the invention provides a method for treating hair loss disorders that can be treated by compounds that modulate (e.g., inhibit) the activity of a JAK (JAK1, JAK2 and/or JAK3). The method comprises administering to a mammalian subject an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), once or twice per day, wherein the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is in the range of about 4 mg/day to about 50 mg/day (such as 4 mg/day to 50 mg/day), for example, about 5 mg/day (such as 5 mg/day), about 10 mg/day (such as 10 mg/day), about 20 mg/day (such as 20 mg/day), about 30 mg/day (such as 30 mg/day), about 40 mg/day (such as 40 mg/day), or about 50 mg/day (such as 50 mg/day).

0038In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt) administered in the method for treating hair loss disorders, is about 4 mg/day (such as 4 mg/day), about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 32 mg/day (such as 32 mg/day) or about 48 mg/day (such as 48 mg/day).

0039In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders, is about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 24 mg/day (such as 24 mg/day), or about 32 mg/day (such as 32 mg/day). In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 8 mg/day (such as 8 mg/day), about 12 mg/day (such as 12 mg/day), about 16 mg/day (such as 16 mg/day) or about 24 mg/day (such as 24 mg/day).

0040In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders, is 10.6 mg/day of Compound (I) phosphate, e.g., administered as a 5.3 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 21.1 mg/day of Compound (I) phosphate, e.g., administered as a 10.5 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 31.6 mg/day of Compound (I) phosphate, e.g., administered as a 15.8 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 42.2 mg/day of Compound (I) phosphate, e.g., administered as a 21.1 mg dose twice daily.

0041In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders is about 4 mg (such as 4 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 5.3 mg (such as 5.3 mg) of the phosphate salt of Compound (I) twice per day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders is about 8 mg (such as 8 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 10.5 mg (such as 10.5 mg) of the phosphate salt of Compound (I) twice per day.

0042In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders is about 12 mg (such as 12 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 15.8 mg (such as 15.8 mg) of the phosphate salt of Compound (I) twice per day. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for treating hair loss disorders is about 16 mg (such as 16 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 21.1 mg (such as 21.1 mg) of the phosphate salt of Compound (I) twice per day. In certain embodiments, the hair loss disorder is alopecia areata. In certain embodiments, the subject is a human. In one embodiment, the subject is a human 6 years of age or older. Preferably, Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), is administered orally at any of the dosages described herein. Preferably, the Compound (I), or a pharmaceutically acceptable salt thereof, is administered orally at any of the dosages described herein in a pharmaceutical formulation which is a tablet.

0043In another aspect, the invention provides a method for inducing hair growth in a subject. The method comprises administering to a mammalian subject an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), once or twice per day, wherein the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is in the range of about 4 mg/day to about 50 mg/day (such as 4 mg/day to 50 mg/day), for example, about 5 mg/day (such as 5 mg/day), about 10 mg/day (such as 10 mg/day), about 20 mg/day (such as 20 mg/day), about 30 mg/day (such as 30 mg/day), about 40 mg/day (such as 40 mg/day), or about 50 mg/day (such as 50 mg/day).

0044In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth, is about 4 mg/day (such as 4 mg/day), about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 32 mg/day (such as 32 mg/day) or about 48 mg/day (such as 48 mg/day).

0045In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth, is about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 24 mg/day (such as 24 mg/day), or about 32 mg/day (such as 32 mg/day). In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth, is about 8 mg/day (such as 8 mg/day), about 12 mg/day (such as 12/mg/day), about 16 mg/day (such as 16 mg/day), or about 24 mg/day (such as 24 mg/day).

0046In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth, is 10.6 mg/day of Compound (I) phosphate, e.g., administered as a 5.3 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 21.1 mg/day of Compound (I) phosphate, e.g., administered as a 10.5 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 31.6 mg/day of Compound (I) phosphate, e.g., administered as a 15.8 mg dose twice daily. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 42.2 mg/day of Compound (I) phosphate, e.g., administered as a 21.1 mg dose twice daily.

0047In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth is about 4 mg (such as 4 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 5.3 mg (such as 5.3 mg) of the phosphate salt of Compound (I) twice per day.

0048In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth is about 8 mg (such as 8 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 10.5 mg (such as 10.5 mg) of the phosphate salt of Compound (I) twice per day.

0049In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth is about 12 mg (such as 12 mg) twice per day. In a specific embodiment, Compound (I) is administered as the about 15.8 mg (such as 15.8 mg) of the phosphate salt of Compound (I) twice per day.

0050In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof administered in the method for inducing hair growth is about 16 mg (such as 16 mg) twice per day. In a specific embodiment, Compound (I) is administered as the about 21.1 mg (such as 21.1 mg) of the phosphate salt of Compound (I) twice per day.

0051In certain embodiments, the subject suffers from a hair loss disorder; in further embodiments, the hair loss disorder is alopecia areata. In certain embodiments, the subject is a human. In one embodiment, the subject is a human 6 years of age or older. Preferably, Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), is administered orally at any of the foregoing dosages. Preferably, the Compound (I), or a pharmaceutically acceptable salt thereof, is administered orally at any of the foregoing dosages in a pharmaceutical formulation which is a tablet.

0052Hair loss disorders include, without limitation, androgenetic alopecia, alopecia areata, telogen effluvium, alopecia totalis, and alopecia universalis.

0053Alopecia areata is an autoimmune disease that results in partial or complete loss of hair on the scalp and body that may affect up to 650,000 Americans at any given time. The scalp is the most commonly affected area, but any hair-bearing site can be affected alone or together with the scalp. Onset of the disease can occur throughout life and affects both women and men. Alopecia areata can be associated with serious psychological consequences, including anxiety and depression. There are currently no drugs approved by the U.S. Food and Drug Administration (FDA) for the treatment of alopecia areata.

0054In a specific embodiment, the condition is alopecia areata in a subject such as a mammalian (e.g., human) patient in need thereof. In certain embodiments, the alopecia areata is moderate to severe alopecia areata (for example, hair loss over at least 30% of the scalp, hair loss over at least 40% of the scalp, or hair loss over at least 50% of the scalp).

0055In one embodiment of any aspect, the compound is administered orally once a day. In other embodiments of any aspect, the compound is administered orally twice per day.

0056Effective doses will also vary, as recognized by those skilled in the art, depending on the diseases treated, the severity of the disease, the route of administration, the sex, age and general health condition of the subject, excipient usage, the possibility of co-usage with other therapeutic treatments such as use of other agents and the judgment of the treating physician.

0057The administration of Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt), can continue for as long as necessary to treat a hair loss disorder, e.g., for one week, two weeks, one month, two months, three months, four months, six months, one year, two years, five years, ten years, or longer.

0058The efficacy of treatment of hair loss disorders such as alopecia areata can be measured in a variety of ways, some of which are known in the art. For example, the "severity of alopecia tool", otherwise known as SALT, is a validated assessment scale - developed by the National Alopecia Areata Foundation working committee - to evaluate the degree of hair loss. See, e.g., <nplcit id="ncit0010" npl-type="s"><text>Olsen EA, Hordinsky MK, Price VH, et al. Alopecia areata investigational assessment guidelines - Part II. J Am Acad Dermatol 2004: 51: 440-447</text></nplcit> (incorporated herein by reference). The SALT score is calculated for a patient by measuring the percentage of hair loss in each of the 4 areas of the scalp and adding the total to achieve a composite score. Hair regrowth is reflected by a decrease in the SALT score. For example, no hair on the scalp would have a SALT score of 100 while complete hair regrowth would be a SALT score of 0. In certain embodiments, methods of treatment as described herein can provide a SALT score improvement of at least 10 points after treatment (for example, from a SALT score of 100 prior to treatment to a SALT score of 90 after treatment). In further embodiments, methods of treatment as described herein can provide a SALT score improvement of at least 20 points, 30 points, 40 points, 50 points, 60 points, 70 points, 80 points, 90 points, or 100 points. In certain embodiments, methods of treatment as described herein can provide after treatment at least a 20% improvement from baseline in the patient's SALT score, or at least a 30% improvement from baseline in the patient's SALT score, or at least a 40% improvement from baseline in the patient's SALT score, or at least a 50% improvement from baseline in the patient's SALT score, or at least a 60% improvement from baseline in the patient's SALT score, or at least a 70% improvement from baseline in the patient's SALT score.

0059In certain embodiments, treatment is continued for a period of at least four weeks, or at least 8 weeks, or at least 12 weeks, or at least 16 weeks, or at least 20 weeks, or at least 24 weeks, or at least 28 weeks, or at least 32 weeks, or at least 36 weeks, or at least 40 weeks, or at least 44 weeks, or at least 48 weeks, or at least 52 weeks.

0060In certain embodiments, Compound (I), or a pharmaceutically acceptable salt thereof, is administered in combination with a second therapeutic agent. Preferably, the second therapeutic agent is an agent useful in the treatment of hair loss disorders or autoimmune conditions, such as inhibitors of JAK1, JAK2, or JAK3, and/or STAT1. Such inhibitors include ruxolitinib, tofacitinib, baricitinib, filgotinib, and the like. Other orally administered second therapeutic agents include agents used in the treatment of alopecia areata, including, for example, oral corticosteroids.

0061For pharmaceutical compositions that comprise a second therapeutic agent, an effective amount of the second therapeutic agent is between about 20% and 100% of the dosage normally utilized in a monotherapy regime using just that agent. Preferably, an effective amount is between about 70% and 100% of the normal monotherapeutic dose. The normal monotherapeutic dosages of these second therapeutic agents are well known in the art. <i>See,</i> e.g., <nplcit id="ncit0011" npl-type="b"><text>Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000</text></nplcit>); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000); the FDA-approved labeling information for ruxolitinib and tofacitinib; and clinical trial information for baricitinib and filgotinib, each of which references are incorporated herein by reference in their entirety.

0062It is expected that some of the second therapeutic agents referenced above will act synergistically with the compounds of this invention. When this occurs, it will allow the effective dosage of the second therapeutic agent and/or Compound (I), or a pharmaceutically acceptable salt thereof, to be reduced from that required in a monotherapy. This has the advantage of minimizing toxic side effects of either the second therapeutic agent or Compound (I), or a pharmaceutically acceptable salt thereof, synergistic improvements in efficacy, improved ease of administration or use and/or reduced overall expense of compound preparation or formulation.

0063In another embodiment, any of the above methods of treatment comprises the further step of co-administering to the subject in need thereof one or more second therapeutic agents. The choice of second therapeutic agent may be made from any second therapeutic agent known to be useful for treatment of hair loss disorders such as alopecia areata. The choice of second therapeutic agent is also dependent upon the particular disease or condition to be treated. Examples of second therapeutic agents that may be employed in the methods of this invention are those set forth above for use in combination compositions comprising Compound (I), or a pharmaceutically acceptable salt thereof, and a second therapeutic agent. Additional therapeutic agents include agents used in the treatment of alopecia areata, including, for example, topical minoxidil, injected corticosteroids, and anthralin cream or ointment.

0064The term "co-administered" as used herein means that the second therapeutic agent may be administered together with Compound (I) or a pharmaceutically acceptable salt thereof, as part of a single dosage form (such as a composition of this invention comprising a compound of the invention and an second therapeutic agent as described above) or as separate, multiple dosage forms. Alternatively, the additional agent may be administered prior to, consecutively with, or following the administration of Compound (I), or a pharmaceutically acceptable salt thereof. In such combination therapy treatment, both Compound (I), or a pharmaceutically acceptable salt thereof, and the second therapeutic agent(s) are administered by conventional methods. The administration of a composition of this invention, comprising both Compound (I), or a pharmaceutically acceptable salt thereof, and a second therapeutic agent, to a subject does not preclude the separate administration of that same therapeutic agent, any other second therapeutic agent or Compound (I), or a pharmaceutically acceptable salt thereof, to said subject at another time during a course of treatment.

0065Effective amounts of these second therapeutic agents are well known to those skilled in the art and guidance for dosing may be found in patents and published patent applications referenced herein, as well as in <nplcit id="ncit0012" npl-type="b"><text>Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000</text></nplcit>); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000), and other medical texts. However, it is well within the skilled artisan's purview to determine the second therapeutic agent's optimal effective-amount range.

0066In one embodiment of the invention, where a second therapeutic agent is administered to a subject, the effective amount of Compound (I), or a pharmaceutically acceptable salt thereof, is less than its effective amount would be where the second therapeutic agent is not administered. In another embodiment, the effective amount of the second therapeutic agent is less than its effective amount would be where Compound (I), or a pharmaceutically acceptable salt thereof, is not administered. In this way, undesired side effects associated with high doses of either agent may be minimized. Other potential advantages (including without limitation improved dosing regimens and/or reduced drug cost) will be apparent to those of skill in the art.

0067In yet another aspect, the invention provides the use of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt), alone or together with one or more of the above-described second therapeutic agents in the manufacture of a medicament, either as a single composition or as separate dosage forms, for treatment or prevention in a subject of a disease, disorder or symptom set forth above. Another aspect of the invention is Compound (I), or a pharmaceutically acceptable salt thereof, for use in the treatment or prevention in a subject of a disease, disorder or symptom thereof delineated herein.

0068Another aspect of the invention is a pharmaceutical composition comprising Compound (I), in the range of about 4 mg to about 50 mg (for example, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg), or an equivalent amount of a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or diluent. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg, 8 mg, 16 mg, 24 mg, 32 mg or 48 mg. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 4 mg, 8 mg, 12 mg, or 16 mg. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 5.3 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 10.5 or 10.6 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 15.8 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 21.1 mg of Compound (I) phosphate. In certain embodiments, the pharmaceutical composition is a tablet.

0069Another aspect of the invention is a unit dose form comprising Compound (I), in the range of about 4 mg to about 50 mg (for example, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg), or an equivalent amount of a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier or diluent. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is about 4 mg, 8 mg, 16 mg, 24 mg, 32 mg or 48 mg. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 4 mg, 8 mg, 12 mg, or 16 mg. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 5.3 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 10.5 or 10.6 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 15.8 mg of Compound (I) phosphate. In certain embodiments, the amount of Compound (I), or a pharmaceutically acceptable salt thereof, is 21.1 mg of Compound (I) phosphate. In certain embodiments, the unit dose form is a tablet.

0070In one embodiment, any atom not designated as deuterium is present at its natural isotopic abundance in Compound (I), or a pharmaceutically acceptable salt thereof.

0071The synthesis of Compound (I), or a pharmaceutically acceptable salt thereof (such as the phosphate salt) may be readily achieved by the methods described <patcit id="pcit0008" dnum="US9249149B"><text>U.S. Patent No. 9,249,149</text></patcit>, the teachings of which are incorporated herein by reference, with appropriate modifications. For example, <patcit id="pcit0009" dnum="US9249149B"><text>U.S. Patent No. 9,249,149</text></patcit> describes the use of a D9-intermediate <b>15</b> to produce a D9-ruxolitinib product; use of the intermediate <b>A</b><chemistry id="chem0002" num="0002"><img file="EP4424367A2_D0002.tif" /></chemistry> in the methods described in <patcit id="pcit0010" dnum="US9249149B"><text>U.S. Patent No. 9,249,149</text></patcit> furnishes Compound (I). Additionally, intermediate <b>B</b><chemistry id="chem0003" num="0003"><img file="EP4424367A2_D0003.tif" /></chemistry> can be used instead of intermediate <b>14</b> of <patcit id="pcit0011" dnum="US9249149B"><text>U.S. Patent No. 9,249,149</text></patcit> to prepare Compound (I); removal of the amino protecting group can be accomplished with basic cleavage (e.g., with sodium hydroxide). Phosphoric acid can be used to convert Compound (I) (Free base) to its phosphate salt. Additional methods of preparing ruxolitinib (i.e., non-deuterated Compound (I)) are disclosed in <patcit id="pcit0012" dnum="US9000161B"><text>U.S. Patent No. 9,000,161</text></patcit>, and can be used, with use of suitable deuterated reagents, to prepare Compound (I).

0072Such methods can be carried out utilizing corresponding deuterated and optionally, other isotope-containing reagents and/or intermediates to synthesize the compounds delineated herein, or invoking standard synthetic protocols known in the art for introducing isotopic atoms to a chemical structure.

0073The invention also provides pharmaceutical compositions comprising an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt); and a pharmaceutically acceptable carrier. The carrier(s) are "acceptable" in the sense of being compatible with the other ingredients of the formulation and, in the case of a pharmaceutically acceptable carrier, not deleterious to the recipient thereof in an amount used in the medicament. In certain embodiments, the pharmaceutical composition is provided as a unit dose form.

0074The invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and 4 to 50 mg of a compound represented by the following structural formula: <chemistry id="chem0004" num="0004"><img file="EP4424367A2_D0004.tif" /></chemistry> or a pharmaceutically acceptable salt thereof (i.e., an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt).

0075Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

0076If required, the solubility and bioavailability of the compounds of the present invention in pharmaceutical compositions may be enhanced by methods well-known in the art. One method includes the use of lipid excipients in the formulation. See "<nplcit id="ncit0013" npl-type="b"><text>Oral Lipid-Based Formulations: Enhancing the Bioavailability of Poorly Water-Soluble Drugs (Drugs and the Pharmaceutical Sciences)," David J. Hauss, ed. Informa Healthcare, 2007</text></nplcit>; and "<nplcit id="ncit0014" npl-type="b"><text>Role of Lipid Excipients in Modifying Oral and Parenteral Drug Delivery: Basic Principles and Biological Examples," Kishor M. Wasan, ed. Wiley-Interscience, 2006</text></nplcit>.

0077Another known method of enhancing bioavailability is the use of an amorphous form of a compound of this invention optionally formulated with a poloxamer, such as LUTROL^™ and PLURONIC^™ (BASF Corporation), or block copolymers of ethylene oxide and propylene oxide. See <patcit id="pcit0013" dnum="US7014866B"><text>United States patent 7,014,866</text></patcit>; and <patcit id="pcit0014" dnum="US20060094744A"><text>United States patent publications 20060094744 </text></patcit>and <patcit id="pcit0015" dnum="US20060079502A"><text>20060079502</text></patcit>.

0078The pharmaceutical compositions of the invention include those suitable for oral administration. Other formulations may conveniently be presented in unit dosage form, e.g., tablets, sustained release capsules, granules, and in liposomes, and may be prepared by any methods well known in the art of pharmacy. See, for example,<nplcit id="ncit0015" npl-type="b"><text> Remington: The Science and Practice of Pharmacy, Lippincott Williams &amp; Wilkins, Baltimore, MD (20th ed. 2000</text></nplcit>).

0079Such preparative methods include the step of bringing into association with the molecule to be administered ingredients such as the carrier that constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers, liposomes or finely divided solid carriers, or both, and then, if necessary, shaping the product.

0080In certain embodiments, the compound is administered orally. Compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, sachets, or tablets each containing a predetermined amount of the active ingredient; a powder or granules; a solution or a suspension in an aqueous liquid or a non-aqueous liquid; an oil-in-water liquid emulsion; a water-in-oil liquid emulsion; packed in liposomes; or as a bolus, etc. Soft gelatin capsules can be useful for containing such suspensions, which may beneficially increase the rate of compound absorption. In a specific embodiment, the compound is administered orally as a tablet.

0081In the case of tablets for oral use, carriers that are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added. In another embodiment, the composition is in the form of a tablet. In certain embodiments, exemplary formulations for the tablet are disclosed in <patcit id="pcit0016" dnum="US8754224B"><text>US. Patent No. 8,754,224</text></patcit>, the teachings of which are herein incorporated by reference.

0082In a particular embodiment, a tablet formulation contains about 4 mg to about 50 mg of Compound (I), or an equivalent amount of a pharmaceutically acceptable salt thereof (such as the phosphate salt), and the following inactive ingredients: colloidal silicon dioxide, magnesium stearate, microcrystalline cellulose, and povidone. Wet granulation followed by compression provides tablets comprising Compound (I), or a pharmaceutically acceptable salt thereof. For example, to prepare a 200 mg tablet comprising the equivalent of 16 mg of Compound (I), 10.6 wt% of Compound (I) phosphate and 64.44 wt% Avicel PH-101 microcrystalline cellulose are mixed in a higher shear granulator, and an 8.5% w/w aqueous Kollidon 30 solution (containing Kollidon 30, a polyvinylpyrrolidone (povidone); 5 wt% (based on the total formulation weight) is added during mixing to form granules. The granules are tray-dried in an oven at 60±10°C and milled using a Quadro Comil U5 mill. The granules retained on the comil screen are forced through a #20 mesh sieve using a stainless steel spatula. The resulting milled granules are mixed with Avicel PH-200 microcrystalline cellulose (18.5 wt%), Aerosil 200 colloidal silicon dioxide (0.5 wt%) and Hyqual magnesium stearate (1 wt%) in a Turbula mixer to form the final blend. The final blend is compressed into 200 mg tablets using a Riva Piccola rotary press tooled with 0.451" × 0.229" D-type modified capsule shape tooling. Each tablet contains 21.1 mg Compound (I) (equivalent to 16 mg of Compound (I) free base).

0083In a particular embodiment, the tablet contains about 10.5 mg or about 10.6 mg of the phosphate salt of Compound (I) (equivalent to 8 mg of Compound (I) free base).

0084In a particular embodiment, the tablet comprises the following ingredients: <tables id="tabl0001" num="0001"><table frame="all"><title><b>4 mg Tablet</b></title><tgroup cols="4"><colspec colnum="1" colname="col1" colwidth="42mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><thead valign="top"><row><entry>Component</entry><entry>Function</entry><entry align="center">Wt %</entry><entry align="center">Amount per unit (mg)</entry></row></thead><tbody><row><entry>Compound (I) Phosphate</entry><entry>Active</entry><entry align="center">2.6</entry><entry align="center">5.3*</entry></row><row><entry>Microcrystalline Cellulose</entry><entry>Diluent/Binder</entry><entry align="center">90.9</entry><entry align="center">181.7</entry></row><row><entry>Povidone</entry><entry>Binder</entry><entry align="center">5.0</entry><entry align="center">10.0</entry></row><row><entry>Colloidal Silicon Dioxide</entry><entry>Glidant</entry><entry align="center">0.5</entry><entry align="center">1.0</entry></row><row><entry>Magnesium Stearate</entry><entry>Lubricant</entry><entry align="center">1.0</entry><entry align="center">2.0</entry></row><row><entry>Purified Water</entry><entry>Solvent</entry><entry namest="col3" nameend="col4" align="center">Removed during processing</entry></row><row><entry namest="col1" nameend="col2" align="right">Total</entry><entry align="center">100.0</entry><entry align="center">200.0</entry></row></tbody></tgroup><tgroup cols="4" rowsep="0"><colspec colnum="1" colname="col1" colwidth="42mm" align="justify" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><tbody><row><entry namest="col1" nameend="col4">*Equivalent to 4 mg Compound (I) free base</entry></row></tbody></tgroup></table></tables>

0085In another particular embodiment, the tablet comprises the following ingredients: <tables id="tabl0002" num="0002"><table frame="all"><title><b>8 mg Tablet</b></title><tgroup cols="4"><colspec colnum="1" colname="col1" colwidth="42mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><thead valign="top"><row><entry>Component</entry><entry>Function</entry><entry align="center">Wt %</entry><entry align="center">Amount per unit (mg)</entry></row></thead><tbody><row><entry>Compound (I) Phosphate</entry><entry>Active</entry><entry align="center">5.2</entry><entry align="center">10.5*</entry></row><row><entry>Microcrystalline Cellulose</entry><entry>Diluent/Binder</entry><entry align="center">90.8</entry><entry align="center">181.5</entry></row><row><entry>Povidone</entry><entry>Binder</entry><entry align="center">2.5</entry><entry align="center">5.0</entry></row><row><entry>Colloidal Silicon Dioxide</entry><entry>Glidant</entry><entry align="center">0.5</entry><entry align="center">1.0</entry></row><row><entry>Magnesium Stearate</entry><entry>Lubricant</entry><entry align="center">1.0</entry><entry align="center">2.0</entry></row><row><entry>Purified Water</entry><entry>Solvent</entry><entry namest="col3" nameend="col4" align="center">Removed during processing</entry></row><row><entry namest="col1" nameend="col2" align="right">Total</entry><entry align="center">100.0</entry><entry align="center">200.0</entry></row></tbody></tgroup><tgroup cols="4" rowsep="0"><colspec colnum="1" colname="col1" colwidth="42mm" align="justify" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><tbody><row><entry namest="col1" nameend="col4">*Equivalent to 8 mg Compound (I) free base</entry></row></tbody></tgroup></table></tables>

0086In an alternative particular embodiment, the tablet comprises the following ingredients: <tables id="tabl0003" num="0003"><table frame="all"><title><b>8 mg Tablet</b></title><tgroup cols="4"><colspec colnum="1" colname="col1" colwidth="42mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><thead valign="top"><row><entry>Component</entry><entry>Function</entry><entry align="center">Wt %</entry><entry align="center">Amount per unit (mg)</entry></row></thead><tbody><row><entry>Compound (I) Phosphate</entry><entry>Active</entry><entry align="center">5.3</entry><entry align="center">10.6*</entry></row><row><entry>Microcrystalline Cellulose</entry><entry>Diluent/Binder</entry><entry align="center">88.2</entry><entry align="center">176.4</entry></row><row><entry>Povidone</entry><entry>Binder</entry><entry align="center">5.0</entry><entry align="center">10.0</entry></row><row><entry>Colloidal Silicon Dioxide</entry><entry>Glidant</entry><entry align="center">0.5</entry><entry align="center">1.0</entry></row><row><entry>Magnesium Stearate</entry><entry>Lubricant</entry><entry align="center">1.0</entry><entry align="center">2.0</entry></row><row><entry>Purified Water</entry><entry>Solvent</entry><entry namest="col3" nameend="col4" align="center">Removed during processing</entry></row><row><entry namest="col1" nameend="col2" align="right">Total</entry><entry align="center">100.0</entry><entry align="center">200.0</entry></row></tbody></tgroup><tgroup cols="4" rowsep="0"><colspec colnum="1" colname="col1" colwidth="42mm" align="justify" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><tbody><row><entry namest="col1" nameend="col4">*Equivalent to 8 mg Compound (I) free base</entry></row></tbody></tgroup></table></tables>

0087In still another particular embodiment, the tablet comprises the following ingredients: <tables id="tabl0004" num="0004"><table frame="all"><title><b>16 mg Tablet</b></title><tgroup cols="4"><colspec colnum="1" colname="col1" colwidth="42mm" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><thead valign="top"><row><entry>Component</entry><entry>Function</entry><entry align="center">Wt %</entry><entry align="center">Amount per unit (mg)</entry></row></thead><tbody><row><entry>Compound (I) Phosphate</entry><entry>Active</entry><entry align="center">10.6</entry><entry align="center">21.1*</entry></row><row><entry>Microcrystalline Cellulose</entry><entry>Diluent/Binder</entry><entry align="center">82.9</entry><entry align="center">165.9</entry></row><row><entry>Povidone</entry><entry>Binder</entry><entry align="center">5.0</entry><entry align="center">10.0</entry></row><row><entry>Colloidal Silicon Dioxide</entry><entry>Glidant</entry><entry align="center">0.5</entry><entry align="center">1.0</entry></row><row><entry>Magnesium Stearate</entry><entry>Lubricant</entry><entry align="center">1.0</entry><entry align="center">2.0</entry></row><row><entry>Purified Water</entry><entry>Solvent</entry><entry namest="col3" nameend="col4" align="center">Removed during processing</entry></row><row><entry namest="col1" nameend="col2" align="right">Total</entry><entry align="center">100.0</entry><entry align="center">200.0</entry></row></tbody></tgroup><tgroup cols="4" rowsep="0"><colspec colnum="1" colname="col1" colwidth="42mm" align="justify" /><colspec colnum="2" colname="col2" colwidth="26mm" /><colspec colnum="3" colname="col3" colwidth="21mm" /><colspec colnum="4" colname="col4" colwidth="34mm" /><tbody><row><entry namest="col1" nameend="col4">*Equivalent to 16 mg Compound (I) free base</entry></row></tbody></tgroup></table></tables>

0088In another embodiment, a composition of this invention further comprises a second therapeutic agent. The second therapeutic agent may be selected from any compound or therapeutic agent known to have or that demonstrates advantageous properties when administered with a compound having the same mechanism of action as ruxolitinib.

0089Preferably, the second therapeutic agent is an agent useful in the treatment of hair loss disorders or autoimmune conditions, including inhibitors of JAK1, JAK2, or JAK3, and/or STAT1. Such inhibitors include ruxolitinib, tofacitinib, baricitinib, filgotinib, and the like. Other second therapeutic agents include oral corticosteroids.

0090In another embodiment, the invention provides separate dosage forms of Compound (I), or a pharmaceutically acceptable salt thereof, and one or more of any of the above-described second therapeutic agents, wherein Compound (I), or a pharmaceutically acceptable salt thereof, and second therapeutic agent are associated with one another. The term "associated with one another" as used herein means that the separate dosage forms are packaged together or otherwise attached to one another such that it is readily apparent that the separate dosage forms are intended to be sold and administered together (within less than 24 hours of one another, consecutively or simultaneously).

0091In the pharmaceutical compositions of the invention, Compound (I), or a pharmaceutically acceptable salt thereof, is present in an effective amount. As used herein, the term "effective amount" refers to an amount which, when administered in a proper dosing regimen, is sufficient to treat the target disorder.

0092The interrelationship of dosages for animals and humans (based on milligrams per meter squared of body surface) is described in<nplcit id="ncit0016" npl-type="s"><text> Freireich et al., Cancer Chemother. Rep, 1966, 50: 219</text></nplcit>. Body surface area may be approximately determined from height and weight of the subject. See, e.g., <nplcit id="ncit0017" npl-type="b"><text>Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 1970, 537</text></nplcit>.

0093In one embodiment, an effective amount of Compound (I) (either as the free base, or as an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt) can range from about 4 mg to 50 mg per day (such as 4 mg to 50 mg per day), such as, about 5 mg/day (such as 5 mg/day), about 10 mg/day (such as 10 mg/day), about 20 mg/day (such as 20 mg/day), about 30 mg/day (such as 30 mg/day), about 40 mg/day (such as 40 mg/day), or about 50 mg/day (such as 50 mg/day). In certain embodiments, the amount is about 4 mg/day (such as 4 mg/day), about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 24 mg/day (such as 24 mg/day), about 32 mg/day (such as 32 mg/day) or about 48 mg/day (such as 48 mg/day). In one embodiment, a dose of about 4 mg/day (such as 4 mg/day), about 8 mg/day (such as 8 mg/day), about 16 mg/day (such as 16 mg/day), about 24 mg/day (such as 24 mg/day), about 32 mg/day (such as 32 mg/day) or about 48 mg/day (such as 48 mg/day) is administered once a day. In a specific example, a dose of 16 mg/day is administered as two 8 mg tablets of Compound (I) (either as the free base, or as an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt) administered together (i.e., as a single dose). In another specific example, a dose of 16 mg/day is administered as one 16 mg tablet of Compound (I) (either as the free base, or as an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt). In another embodiment, a dose of 4 mg/day, 8 mg/day, 16 mg/day, 24 mg/day, 32 mg/day or 48 mg/day is administered in divided doses, twice a day (e.g., a 48 mg/day dose is administered as 24 mg twice daily). In another embodiment, a dose of 8 mg/day, 16 mg/day, 24 mg/day, or 32 mg/day is administered in divided doses, twice a day (e.g., a 32 mg/day dose is administered as 16 mg of Compound (I) (either as the free base, or as an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt) twice daily, i.e., in separate doses. In one specific embodiment, a dose of 16 mg/day is administered as 8 mg of Compound (I) (either as the free base, or as an equivalent amount of a pharmaceutically acceptable salt, such as the phosphate salt) twice daily, i.e., in separate doses. It will be understood that reference to an amount of Compound (I), or a pharmaceutically acceptable salt thereof, includes an amount of a pharmaceutically acceptable salt of Compound (I) (such as the phosphate salt) which is equivalent to the stated amount of Compound (I) as the free base (e.g., 10.5 mg of Compound (I) phosphate salt is equivalent to 8 mg of Compound (I) free base).

0094In certain embodiments, an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof is about 4 mg (such as 4 mg) twice per day. In a specific embodiment, an effective amount of Compound (I) is administered as about 5.3 mg (such as 5.3 mg) of the phosphate salt of Compound (I) twice per day. In certain embodiments, an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof is about 8 mg (such as 8 mg) twice per day. In a specific embodiment, Compound (I) is administered as about 10.5 mg (such as 10.5 mg) of the phosphate salt of Compound (I) twice per day.

0095In certain embodiments, an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof is about 12 mg (such as 12 mg) twice per day. In a specific embodiment, effective amount of Compound (I) is about 15.8 mg (such as 15.8 mg) of the phosphate salt of Compound (I) twice per day. In certain embodiments, an effective amount of Compound (I), or a pharmaceutically acceptable salt thereof is about 16 mg (such as 16 mg) twice per day. In a specific embodiment, the effective amount of Compound (I) is about 21.1 mg (such as 21.1 mg) of the phosphate salt of Compound (I) twice per day.

Examples

Example 1. Determination of Metabolic Stability of D-Ruxolitinib using CYP3A4 Supersomes

Materials and Methods:

0096<b><i>Materials:</i></b> CYP3A4 supersomes^™ were obtained from Corning Gentest. β-nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), magnesium chloride (MgClz), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich. Deuterated test compounds were supplied by Concert Pharmaceuticals.

0097<b><i>Determination of Metabolic Stability:</i></b> 10 mM stock solutions of test compounds were prepared in DMSO. The 7.5 mM stock solutions were diluted to12.75 µM in acetonitrile (ACN). The CYP3A4 supersomes were diluted in 0.1 M potassium phosphate buffer, pH 7.4, containing 3 mM MgCl₂. The diluted supersomes were added to wells of a 96-well deep-well polypropylene plate in triplicate. A 10 µL aliquot of the 12.75 µM test compound was added to the supersomes and the mixture was pre-warmed for 10 minutes. Reactions were initiated by addition of pre-warmed NADPH solution. The final reaction volume was 0.5 mL and contained 50 pmol/mL CYP3A4 supersomes, 0.25 µM test compound, and 2 mM NADPH in 0.1 M potassium phosphate buffer, pH 7.4, and 3 mM MgCl₂. The reaction mixtures were incubated at 37°C and 50 µL aliquots were removed at 0, 5, 10, 20, and 30 minutes and added to shallow-well 96-well plates which contained 50 µL of ice-cold ACN with internal standard to stop the reactions. The plates were stored at 4°C for 20 minutes after which 100 µL of water was added to the wells of the plate before centrifugation to pellet precipitated proteins. Supernatants were transferred to another 96-well plate and analyzed for amounts of parent remaining by LC-MS/MS using an Applied Bio-systems mass spectrometer.

0098<b><i>Data analysis:</i></b> The <i>in vitro</i> t½s for test compounds were calculated from the slopes of the linear regression of % parent remaining (ln) vs incubation time relationship. <maths id="math0001"><math display="block"><mi>in</mi><mspace width="1ex" /><mi>vitro</mi><mspace width="1ex" /><msub><mi mathvariant="normal">t</mi><mstyle scriptlevel="+1"><mfrac bevelled="true"><mn>1</mn><mn>2</mn></mfrac></mstyle></msub><mo>=</mo><mn>0.693</mn><mo>/</mo><mi mathvariant="normal">k</mi></math><img file="EP4424367A2_D0005.tif" /></maths> k = -[slope of linear regression of % parent remaining(ln) vs incubation time] Data analysis was performed using Microsoft Excel Software.

0099It was found that Compound (I) had a t½, about 80% longer than the t½ of non-deuterated ruxolitinib. These results show that Compound (I) is substantially more stable metabolically than ruxolitinib in the CYP3A4 supersome assay.

Example 2. Determination of Metabolic Stability of D-Ruxolitinib using Human Liver Microsomes

0100<b><i>Materials:</i></b> Human liver microsomes (20 mg/mL) were obtained from Xenotech, LLC (Lenexa, KS). β-nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), magnesium chloride (MgClz), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich. Deuterated test compounds were supplied by Concert Pharmaceuticals.

0101<b><i>Determination of Metabolic Stability:</i></b> 7.5 mM stock solutions of test compounds were prepared in DMSO. The 7.5 mM stock solutions were diluted to12.5 µM in acetonitrile (ACN). The human liver microsomes were diluted in 0.1 M potassium phosphate buffer, pH 7.4, containing 3 mM MgCl₂. The diluted microsomes were added to wells of a 96-well deep-well polypropylene plate in triplicate. A 10 µL aliquot of the 12.5 µM test compound was added to the microsomes and the mixture was pre-warmed for 10 minutes. Reactions were initiated by addition of pre-warmed NADPH solution. The final reaction volume was 0.5 mL and contained 5 mg/mL human liver microsomes, 0.25 µM test compound, and 2 mM NADPH in 0.1 M potassium phosphate buffer, pH 7.4, and 3 mM MgCl₂. The reaction mixtures were incubated at 37°C, and 50 µL aliquots were removed at 0, 5, 10, 20, and 30 minutes and added to shallow-well 96-well plates which contained 50 µL of ice-cold ACN with internal standard to stop the reactions. The plates were stored at 4°C for 20 minutes after which 100 µL of water was added to the wells of the plate before centrifugation to pellet precipitated proteins. Supernatants were transferred to another 96-well plate and analyzed for amounts of parent remaining by LC-MS/MS using an Applied Bio-systems mass spectrometer.

0102<b><i>Data analysis:</i></b> The <i>in vitro</i> t½s for test compounds were calculated from the slopes of the linear regression of % parent remaining (ln) vs incubation time relationship. <maths id="math0002"><math display="block"><mi>in</mi><mspace width="1ex" /><mi>vitro</mi><mspace width="1ex" /><msub><mi mathvariant="normal">t</mi><mstyle scriptlevel="+1"><mfrac bevelled="true"><mn>1</mn><mn>2</mn></mfrac></mstyle></msub><mo>=</mo><mn>0.693</mn><mo>/</mo><mi mathvariant="normal">k</mi></math><img file="EP4424367A2_D0006.tif" /></maths> k = -[slope of linear regression of % parent remaining(ln) vs incubation time] Data analysis was performed using Microsoft Excel Software.

0103It was found that Compound (I) had a t½ about 75% longer than the t½ of non-deuterated ruxolitinib. These results show that Compound (I) is substantially more stable metabolically than ruxolitinib in the HLM assay.

Example 3

-

Human Studies

0104Single Ascending Dose (SAD) Study: Healthy volunteers were administered doses of Compound (I) or placebo under fasted conditions. The objective of the study was to evaluate the pharmacokinetics of a single dose of 8 mg, 16 mg, 32 mg, or 48 mg of Compound (I) as the phosphate salt (e.g., 10.5 mg of Compound (I) phosphate salt is equivalent to 8 mg of Compound (I) free base). For each dose group, 6 subjects received Compound (I), and 2 subjects received placebo. The study design is shown in <figref idref="f0001">FIG. 1A</figref>. Metabolites of Compound (I) were analyzed. Doses of Compound (I) were administered as Compound (I) phosphate salt as a powder-in-capsule, administered with water. The preliminary results are shown in <figref idref="f0001">FIGS. 1B</figref> and <figref idref="f0002">1C</figref>.

0105Multiple Ascending Dose Study: Healthy volunteers were administered doses of Compound (I) or placebo under fasted conditions. The objective of the study was to evaluate the pharmacokinetics of daily administration of 8 mg, 16 mg, 24 mg, or 32 mg of Compound (I) as the phosphate salt (e.g., 10.5 mg of Compound (I) phosphate salt is equivalent to 8 mg of Compound (I) free base). Compound (I) was administered once per day (QD) (8 mg, 24 mg, 32 mg doses) or twice per day (BID) (two 8 mg doses totaling 16 mg/day, or two 16 mg doses totaling 32 mg/day) for seven consecutive days. For each dose group, 8 subjects received Compound (I), and 2 subjects received placebo. The study design is shown in <figref idref="f0001">FIG. 1A</figref>. Metabolites of Compound (I) were analyzed. Doses of Compound (I) were administered as 8 mg tablets, with water. The preliminary results are shown in <figref idref="f0001">FIGS.1B</figref> and <figref idref="f0002">1C</figref>.

0106Final results are shown in the following table: <tables id="tabl0005" num="0005"><table frame="all"><tgroup cols="9"><colspec colnum="1" colname="col1" colwidth="21mm" /><colspec colnum="2" colname="col2" colwidth="17mm" /><colspec colnum="3" colname="col3" colwidth="17mm" /><colspec colnum="4" colname="col4" colwidth="17mm" /><colspec colnum="5" colname="col5" colwidth="22mm" /><colspec colnum="6" colname="col6" colwidth="22mm" /><colspec colnum="7" colname="col7" colwidth="19mm" /><colspec colnum="8" colname="col8" colwidth="17mm" /><colspec colnum="9" colname="col9" colwidth="17mm" /><thead valign="top"><row><entry align="center" /><entry align="center"><b>Median (range)</b></entry><entry namest="col3" nameend="col9" align="center"><b>Arithmetic Mean (CV%)</b></entry></row><row><entry align="right"><b>Treatment</b></entry><entry align="center"><b>T_max (hr)</b></entry><entry align="center"><b>C_max (ng/mL)</b></entry><entry align="center"><b>t_1/2 (hr)</b></entry><entry align="center"><b>AUC_{0-24 hr} (hr*ng/mL)</b></entry><entry align="center"><b>AUC_0-inf (hr*ng/mL)</b></entry><entry align="center"><b>C_24hr (ng/mL)</b></entry><entry align="center"><b>CL/F (L/hr)</b></entry><entry align="center"><b>Vz/F (L)</b></entry></row></thead><tbody><row><entry align="right">CTP-543 8 mg (n=6)</entry><entry align="center">1.25 (0.50 - 1.50)</entry><entry align="center">118 (28.4%)</entry><entry align="center">3.14 (28.4%)</entry><entry align="center">584 (31.3%)</entry><entry align="center">589 (31.6%)</entry><entry align="center">1.02 (94.9%)</entry><entry align="center">14.92 (34.9%)</entry><entry align="center">64.1 (27.8%)</entry></row><row><entry align="right">CTP-543 16 mg (n=6)</entry><entry align="center">1.50 (0.50 - 3.20)</entry><entry align="center">260 (39.2%)</entry><entry align="center">3.32 (25.9%)</entry><entry align="center">1329 (34.7%)</entry><entry align="center">1347 (35.2%)</entry><entry align="center">2.77 (78.4%)</entry><entry align="center">13.87 (52.4%)</entry><entry align="center">61.0 (28.0%)</entry></row><row><entry align="right">CTP-543 32mg (n=6)</entry><entry align="center">1.25 (0.50 - 2.00)</entry><entry align="center">535 (29.0%)</entry><entry align="center">3.52 (15.9%)</entry><entry align="center">2424 (25.2%)</entry><entry align="center">2462 (26.3%)</entry><entry align="center">5.60 (104.5%)</entry><entry align="center">13.88 (30.2%)</entry><entry align="center">68.4 (21.3%)</entry></row><row><entry align="right">CTP-543 48mg (n=5)</entry><entry align="center">1.50 (0.50 - 2.00)</entry><entry align="center">853 (25.7%)</entry><entry align="center">3.61 (21.4%)</entry><entry align="center">4095 (34.2%)</entry><entry align="center">4159 (35.1%)</entry><entry align="center">9.26 (80.1%)</entry><entry align="center">12.75 (34.1%)</entry><entry align="center">63.2 (25.6%)</entry></row></tbody></tgroup></table></tables>

0107Preliminary results from the SAD study are shown in <figref idref="f0001">FIGS. 1B</figref> and <figref idref="f0002">1C</figref>. Preliminary results from the MAD study are shown in <figref idref="f0004">FIGS. 2B</figref> and <figref idref="f0005">2C</figref>. Final results from the MAD study are shown in the following tables. In <figref idref="f0001 f0002">FIGS. 1</figref> and <figref idref="f0003 f0004 f0005">2</figref>, CTP-543 is Compound (I), administered as the phosphate salt. <tables id="tabl0006" num="0006"><table frame="all"><tgroup cols="7"><colspec colnum="1" colname="col1" colwidth="32mm" /><colspec colnum="2" colname="col2" colwidth="18mm" /><colspec colnum="3" colname="col3" colwidth="17mm" /><colspec colnum="4" colname="col4" colwidth="22mm" /><colspec colnum="5" colname="col5" colwidth="37mm" /><colspec colnum="6" colname="col6" colwidth="22mm" /><colspec colnum="7" colname="col7" colwidth="23mm" /><thead><row valign="top"><entry namest="col1" nameend="col2" align="center" /><entry align="center"><b>Median (range)</b></entry><entry namest="col4" nameend="col7" align="center"><b>Arithmetic Mean (CV%)</b></entry></row><row><entry align="center"><b>Dose Level of CTP-543</b></entry><entry align="center"><b>Study Day</b></entry><entry align="center" valign="top"><b>T_max (hr)</b></entry><entry align="center" valign="top"><b>C_max (ng/mL)</b></entry><entry align="center" valign="top"><b>AUCtau/ AUC_0-24hr (hr*ng/mL)</b></entry><entry align="center" valign="top"><b>C_24hr (ng/mL)</b></entry><entry align="center" valign="top"><b>t_1/2 (hr)</b></entry></row></thead><tbody><row><entry morerows="1" align="center">8 mg QD (n=7)</entry><entry align="center" valign="middle">Day 1</entry><entry align="center">1.00 (0.50 - 2.00)</entry><entry align="center">156 (31.2%)</entry><entry align="center">704 (26.4%)</entry><entry align="center">0.86 (45.3%)</entry><entry align="center">3.23 (9.6%)</entry></row><row><entry align="center" valign="middle">Day 7</entry><entry align="center">1.00 (0.25 - 1.50)</entry><entry align="center">151 (34.1%)</entry><entry align="center">668 (27.8%)</entry><entry align="center">1.01 (75.0%)</entry><entry align="center">3.51 (20.9%)</entry></row><row><entry morerows="1" align="center">24 mg QD (n=8)</entry><entry align="center" valign="middle">Day 1</entry><entry align="center">0.50 (0.25 - 2.00)</entry><entry align="center">384 (22.2%)</entry><entry align="center">1672 (25.7%)</entry><entry align="center">2.48 (134%)</entry><entry align="center">3.14 (28.8%)</entry></row><row><entry align="center" valign="middle">Day 7</entry><entry align="center">1.00 (0.50 - 2.00)</entry><entry align="center">310 (22.9%)</entry><entry align="center">1534 (25.2%)</entry><entry align="center">3.39 (160%)</entry><entry align="center">3.50 (25.2%)</entry></row><row><entry morerows="1" align="center">32 mg QD (n=8)</entry><entry align="center" valign="middle">Day 1</entry><entry align="center">0.75 (0.50 - 2.00)</entry><entry align="center">509 (32.8%)</entry><entry align="center">2219 (36.2%)</entry><entry align="center">2.95 (141%)</entry><entry align="center">2.86 (32.1%)</entry></row><row><entry align="center" valign="middle">Day 7</entry><entry align="center">0.75 (0.50 - 2.00)</entry><entry align="center">492 (18.0%)</entry><entry align="center">2104 (27.8%)</entry><entry align="center">5.12 (153%)</entry><entry align="center">3.45 (37.1%)</entry></row><row><entry morerows="1" align="center" valign="middle">8 mg BID (n=4)</entry><entry align="center" valign="middle">Day 1</entry><entry align="center">1.00 (0.50 - 1.50)</entry><entry align="center">160 (17.5%)</entry><entry align="center">703 (24.5%)</entry><entry align="center">13.7 (55.9%)</entry><entry align="center">3.77 (23.4%)</entry></row><row><entry align="center" valign="middle">Day 7</entry><entry align="center">1.50 (0.25 - 2.00)</entry><entry align="center">183 (25.7%)</entry><entry align="center">844 (39.8%)</entry><entry align="center">13.2 (59.6%)</entry><entry align="center">4.20 (32.3%)</entry></row><row><entry morerows="1" align="center" valign="middle">16 mg BID (n=8)</entry><entry align="center" valign="middle">Day 1</entry><entry align="center">1.00 (0.50 - 2.00)</entry><entry align="center">290 (19.7%)</entry><entry align="center">1062 (22.5%)</entry><entry align="center">15.3 (53.9%)</entry><entry align="center">2.89 (31.4%)</entry></row><row><entry align="center" valign="middle">Day 7</entry><entry align="center">0.75 (0.50 - 2.00)</entry><entry align="center">288 (36.3%)</entry><entry align="center">1127 (26.6%)</entry><entry align="center">16.5 (71.6%)</entry><entry align="center">3.53 (19.3%)</entry></row></tbody></tgroup></table></tables>

0108In the combined SAD and MAD studies, a total of 77 subjects were dosed (60 received CTP-543; 17 received placebo). CTP-543 was rapidly absorbed and did not accumulate with repeated dosing. No serious adverse events were reported; the most common adverse event reported was headache. No withdrawal or dose modification related to CTP-543 occurred. Cases of mild neutropenia resolved or trended toward recovery after dosing completion. Severe neutropenia (Grade 3 or 4) was not observed.

0109In the Phase 1 study, Compound (I) was generally well tolerated. Based on these studies, doses of 8 mg BID (twice daily), 12 mg BID, 16 mg BID and 32 mg BID were selected.

0110Additional findings from the Phase 1 study show that the average systemic exposure of 16 mg BID of Compound (I), the highest dose being evaluated in the Phase 2a trial described below, appears comparable to published findings for average reported exposure of the 20 mg BID ruxolitinib dose shown to be effective at inducing hair regrowth in patients with moderate to severe alopecia. (See, e.g., <nplcit id="ncit0018" npl-type="s"><text>JCI Insight. 2016;1(15):e89790. doi:10.1172/jci.insight.89790</text></nplcit>.)

0111Pharmacodynamic analyses were also performed during the MAD study of the Phase 1 trial to assess the inhibition of IL-6- and IFN-γ-mediated JAK/STAT signaling. Consistent with the established pharmacological activity of CTP-543 (Compound (I)), a dose-related reduction in IL-6-stimulated phosphorylated STAT3 (pSTAT3) was observed. In general, pSTAT3 inhibition in all treatment groups returned to approximate baseline values at 24 hours post-dose. Also, IFN-γ-mediated STAT1 signaling, which is believed to play a key role in the pathogenesis of alopecia areata, was significantly inhibited in disease-relevant immune cell types at all doses evaluated.

0112A Phase 2a trial has been designed to evaluate the safety and efficacy of Compound (I) after 12 months of dosing with the primary efficacy analysis at week 24. The Phase 2a trial is a double-blind, randomized, placebo-controlled, parallel dose trial to evaluate the safety and efficacy of Compound (I) in adult patients with moderate-to-severe alopecia areata. Approximately 100 patients will be randomized to receive one of four doses of Compound I as the phosphate salt (e.g., 10.5 mg of Compound (I) phosphate salt is equivalent to 8 mg of Compound (I) free base). The four doses of Compound (I) are 4, 8 (i.e., about10.5 mg of Compound (I) phosphate salt), 12 and 16 mg twice daily, and there is also a patient group receiving placebo. The primary outcome measure will utilize the severity of alopecia tool (SALT) after 24 weeks of dosing. The trial will include an additional 28 weeks of dosing where all patients enrolled in the study will receive Compound (I).

0113Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the illustrative examples, make and utilize the compounds of the present invention and practice the claimed methods. It should be understood that the foregoing discussion and examples merely present a detailed description of certain preferred embodiments. It will be apparent to those of ordinary skill in the art that various modifications and equivalents can be made without departing from the spirit and scope of the invention.

0114Further embodiments of the present disclosure include those described in the following, numbered disclosures. <ol id="ol0001"><li>(1) A method of treating a hair loss disorder in a mammalian subject, the method comprising administering to the subject an amount in the range of 4 mg to 50 mg per day, of a compound represented by the following structural formula: <chemistry id="chem0005" num="0005"><img file="EP4424367A2_D0007.tif" /></chemistry><b>Compound (I)</b> , or a pharmaceutically acceptable salt thereof.</li><li>(2) The method of Disclosure 1, wherein the hair loss disorder is alopecia areata.</li><li>(3) The method of any one of Disclosures 1-2, comprising administering to the subject about 5 mg/day, about 10 mg/day, about 20 mg/day, about 30 mg/day, about 40 mg/day, or about 50 mg/day of Compound (I), or a pharmaceutically acceptable salt thereof.</li><li>(4) The method of any one of Disclosures 1-3, comprising administering to the subject about 8 mg/day, about 12 mg/day, about 16 mg/day, or about 24 mg/day of Compound (I), or a pharmaceutically acceptable salt thereof.</li><li>(5) The method of any one of Disclosures 1-4, wherein the compound is administered orally.</li><li>(6) The method of any one of Disclosures 1-5, wherein the compound is administered in a pharmaceutical formulation which is a tablet.</li><li>(7) The method of any one of Disclosures 1-6, wherein the compound is administered once a day.</li><li>(8) The method of any one of Disclosures 1-6, wherein the compound is administered twice a day.</li><li>(9) The method of any one of Disclosures 1-8, wherein in Compound (I), any atom not designated as deuterium is present at its natural isotopic abundance.</li><li>(10) A pharmaceutical composition comprising a pharmaceutically acceptable carrier or diluent and 4 to 50 mg of a compound represented by the following structural formula: <chemistry id="chem0006" num="0006"><img file="EP4424367A2_D0008.tif" /></chemistry> or a pharmaceutically acceptable salt thereof.</li><li>(11) The pharmaceutical composition of Disclosure 10, comprising about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, or about 50 mg of Compound (I), or a pharmaceutically acceptable salt thereof.</li><li>(12) The pharmaceutical composition of Disclosure 10 or 11, wherein the pharmaceutical composition is suitable for oral administration.</li><li>(13) The pharmaceutical composition of Disclosure 10 or 11, wherein the composition is a tablet.</li><li>(14) The pharmaceutical composition of any one of Disclosure 10-13, wherein the composition is administered once a day.</li><li>(15) The pharmaceutical composition of Disclosure 10, wherein the composition is a tablet comprising: <ol id="ol0002" compact="compact"><li>(i) about 5.2 wt% of Compound (I) phosphate salt;</li><li>(ii) about 90.8 wt% microcrystalline cellulose;</li><li>(iii) about 2.5 wt% povidone;</li><li>(iv) about 0.5 wt% colloidal silicon dioxide; and</li><li>(v) about 1.0 wt% magnesium stearate.</li></ol></li><li>(16) The pharmaceutical composition of Disclosure 15, wherein the composition is a 200 mg tablet comprising about 10.5 mg of Compound (I) phosphate salt.</li><li>(17) The pharmaceutical composition of Disclosure 10, wherein the composition is a tablet comprising: <ol id="ol0003" compact="compact"><li>(i) about 5.3 wt% of Compound (I) phosphate salt;</li><li>(ii) about 88.2 wt% microcrystalline cellulose;</li><li>(iii) about 5.0 wt% povidone;</li><li>(iv) about 0.5 wt% colloidal silicon dioxide; and</li><li>(v) about 1.0 wt% magnesium stearate.</li></ol></li><li>(18) The pharmaceutical composition of Disclosure 17, wherein the composition is a 200 mg tablet comprising about 10.6 mg of Compound (I) phosphate salt.</li><li>(19) The pharmaceutical composition of Disclosure 10, wherein the composition is a tablet comprising: <ol id="ol0004" compact="compact"><li>(i) about 10.6 wt% of Compound (I) phosphate salt;</li><li>(ii) about 82.9 wt% microcrystalline cellulose;</li><li>(iii) about 5.0 wt% povidone;</li><li>(iv) about 0.5 wt% colloidal silicon dioxide; and</li><li>(v) about 1.0 wt% magnesium stearate.</li></ol></li><li>(20) The pharmaceutical composition of Disclosure 19, wherein the composition is a 200 mg tablet comprising about 21.1 mg of Compound (I) phosphate salt.</li><li>(21) A method of treating a hair loss disorder in a mammalian subject, the method comprising administering to the subject twice a day about 4 mg of a compound represented by the following structural formula: <chemistry id="chem0007" num="0007"><img file="EP4424367A2_D0009.tif" /></chemistry> or a pharmaceutically acceptable salt thereof.</li><li>(22) The method of Disclosure 21, wherein Compound (I) is administered as about 5.3 mg of the phosphate salt twice per day.</li><li>(23) A method of treating a hair loss disorder in a mammalian subject, the method comprising administering to the subject twice a day about 8 mg of a compound represented by the following structural formula: <chemistry id="chem0008" num="0008"><img file="EP4424367A2_D0010.tif" /></chemistry> or a pharmaceutically acceptable salt thereof.</li><li>(24) The method of Disclosure 23, wherein Compound (I) is administered as about 10.5 mg of the phosphate salt twice per day.</li><li>(25) A method of treating a hair loss disorder in a mammalian subject, the method comprising administering to the subject twice per day about 12 mg of a compound represented by the following structural formula: <chemistry id="chem0009" num="0009"><img file="EP4424367A2_D0011.tif" /></chemistry> or a pharmaceutically acceptable salt thereof.</li><li>(26) The method of Disclosure 25, wherein Compound (I) is administered as about 15.8 mg of the phosphate salt twice per day.</li><li>(27) A method of treating a hair loss disorder in a mammalian subject, the method comprising administering to the subject twice a day about 16 mg of a compound represented by the following structural formula: <chemistry id="chem0010" num="0010"><img file="EP4424367A2_D0012.tif" /></chemistry> or a pharmaceutically acceptable salt thereof.</li><li>(28) The method of Disclosure 27, wherein Compound (I) is administered as about 21.1 mg of the phosphate salt twice per day.</li><li>(29) The method of any one of Disclosures 21-28, wherein the hair loss disorder is alopecia areata.</li><li>(30) The method of any one of Disclosures 21-29, wherein the compound is administered orally.</li><li>(31) The method of any one of Disclosures 21-30 wherein the compound is administered in a pharmaceutical formulation which is a tablet.</li></ol>