KR102261733B1 - Deuterated derivatives of ruxolitinib - Google Patents

Abstract

In one embodiment, the present invention relates to a compound of formula (A) or a pharmaceutically acceptable salt thereof; Pharmaceutical compositions comprising such compounds and methods of treating the diseases disclosed herein are provided:

Description

DEUTERATED DERIVATIVES OF RUXOLITINIB

The present invention relates to novel heteroaryl-substituted pyrrolo[2,3-d]pyrimidines, and pharmaceutically acceptable salts thereof. The present invention also relates to compositions comprising a compound of the present invention and methods of treating diseases and conditions which are advantageously treated by administering an inhibitor of a Janus-associated kinase having selectivity for subtypes 1 and 2 (JAK1/JAK2). of the use of such a composition.

Many current medications suffer from poor absorption, distribution, metabolism and/or excretion (ADME) properties that prevent their widespread use or limit their use in certain regimens. are suffering Poor ADME properties are also a major cause of failure of drug candidates in clinical trials. Although formulation techniques and prodrug strategies can be used to improve certain ADME properties in some cases, these methods often fail to address the underlying ADME problem present in many drugs and drug candidates. . One such problem is the rapid metabolism, which causes many drugs to be eliminated from the body too quickly, which would otherwise be very effective in treating disease. A possible solution to rapid drug clearance is frequent or high dose administration to obtain sufficiently high drug plasma levels. However, these solutions lead to many potential treatment problems, such as poor patient acceptance with dosing regimens, side effects exacerbated by high doses, and increased cost of treatment. Rapidly metabolized drugs may also expose patients to undesirable toxic or reactive metabolites.

Another ADME limitation on which many drugs are affected is the formation of toxic or biologically reactive metabolites. As a result, some patients to whom the drug is administered may experience toxic effects, or the safe dosing of such drugs may be limited, resulting in the patient receiving sub-optimal amounts of the active agent. In certain instances, although varied dosing intervals or formulation methods can help reduce clinical side effects, often the formation of such undesirable metabolites is essential in the metabolism of compounds.

In some select cases, metabolic inhibitors may be co-administered with drugs that are cleared too quickly. Such is the case with drugs in the class of protease inhibitors used to treat HIV infection. The FDA recommends that these drugs be co-administered with ritonavir, an inhibitor of the cytochrome P450 enzyme 3A4 (CYP3A4), an enzyme typically responsible for their metabolism [see, e.g., Kempf, D.J. et al., Antimicrobial agents and chemotherapy, 1997, 41(3): 654-60]. However, ritonavir causes side effects and is added to pills that burden HIV patients who already have to take a combination of several drugs. Similarly, the CYP2D6 inhibitor quinidine has been added to dextromethorphan for the purpose of reducing the rapid CYP2D6 metabolism of dextromethorphan in the treatment of pseudobulbar affect. However, quinidine has undesirable side effects that greatly limit its use in possible combination therapies (Wang, L et al., Clinical Pharmacology and Therapeutics, 1994, 56(6 Pt 1): 659-67; and FDA label for quinidine at www.accessdata.fda.gov].

In general, combining drugs with cytochrome P450 inhibitors is not a satisfactory strategy to reduce drug clearance. Inhibition of CYP enzyme activity can affect the metabolism and clearance of other drugs metabolized by those same enzymes. CYP inhibition can cause other drugs to accumulate in the body at toxic levels.

A potentially attractive strategy for improving the metabolic properties of drugs is deuterium modification. In these methods, attempts have been made to slow the CYP-mediated metabolism of drugs or reduce the formation of undesirable metabolites by replacing one or more hydrogen atoms with deuterium. Deuterium is a safe, stable and non-radioactive isotope of hydrogen. Compared to hydrogen, deuterium forms a stronger bond with carbon. In selected cases, increased binding strength by deuterium can positively affect the ADME properties of a drug, creating the potential for improved drug efficacy, safety and/or tolerability. At the same time, since the size and shape of deuterium is essentially the same as that of hydrogen, the replacement of hydrogen by deuterium would not be expected to affect the biochemical efficacy and selectivity of the drug over the original chemical entity containing only hydrogen. can

Over the past 35 years, the effect of deuterium substitution on metabolic rate has been reported for a very small percentage of approved drugs (Blake, MI et al, J Pharm Sci, 1975, 64:367-91; Foster, AB, Adv Drug Res 1985, 14:1-40 (“Foster”); Kushner, DJ et al, Can J Physiol Pharmacol 1999, 79-88; Fisher, MB et al, Curr Opin Drug Discov Devel, 2006, 9:101-09 (“Fisher”)]. Many examples of these references report the effect of local deuterium isotopes (effect on metabolic rate at specific deuterated sites in the substrate) rather than the effect of deuterium on the overall metabolic stability of the drug, i.e., overall substrate consumption through metabolism. are doing The reported results of these studies measuring the effect of deuterium substitution on overall metabolic safety are varied and unpredictable. For some compounds, deuteration reduced metabolic clearance in vivo. In other cases, there was no change in metabolism. Others also demonstrated increased metabolic clearance. Diversity in deuterium effects also led experts to question or ignore deuterium modification as a viable drug design strategy to suppress side-effect metabolism [Foster at p. 35 and Fisher at p. 101].

The effect of deuterium modification on drug metabolic properties is not predictable even when deuterium atoms are incorporated at known metabolic sites. By actually making and testing a deuterated drug, one can determine whether and how the metabolic rate differs from its non-deuterated counterpart. Literature example [Fukuto et al. (J. Med. Chem. 1991, 34, 2871-76)]. Many drugs have metabolizable ascites sites. The extent of deuteration, if any, required to determine the site where deuterium substitution is required and its effect on metabolism will be different for each drug.

Ruxolitinib phosphate is 3(R)-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile phosphate and (R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-cyclopentylpropanenitrile phosphate is a heteroaryl-substituted pyrrolo[2,3-d]pyrimidine, which inhibits the Janus-associated kinases (JAKs) JAK1 and JAK2. These kinases mediate the signaling of many cytokines and growth factors important for hematopoietic and immune function. JAK signaling involves the recruitment and activation of STATs (signal transducers and transcriptional activators) to cytokine receptors and the subsequent localization of STATs to the nucleus leading to regulation of gene expression.

Ruxoritinib phosphate currently suffers from moderate or high risk myelofibrosis, including primary myelofibrosis, post-polycythemia vera myelofibrosis, and post-essential thrombocythemia. approved for the treatment of patients with Ruxortinib phosphate is also currently used in essential thrombocythemia, pancreatic cancer, prostate cancer, breast cancer, leukemia, non-Hodgkin's lymphoma, multiple myeloma and psoriasis. The treatment is in clinical trials.

The three metabolites in humans are active, one resulting from hydroxylation at the 2-position on the cyclopentyl moiety and one resulting from hydroxylation at the 3-position on the cyclopentyl moiety, and cyclopentyl. It was identified as a ketone resulting from further oxidation at the 3-position on the moiety. See Shilling, A.D. et al., Drug Metabolism and Disposition, 2010, 38(11): 2023-2031; FDA Prescribing Information and US20080312258].

The most common adverse hematological reactions associated with ruxoritinib are thrombocytopenia and anemia. The most common non-hematologic adverse reactions are bruising, dizziness and headache.

Despite the beneficial activity of ruxolitinib, there is a continuing need for new compounds to treat the diseases and conditions mentioned above.

Summary of the invention

The present invention relates to novel heteroaryl-substituted pyrrolo[2,3-d]pyrimidines, and pharmaceutically acceptable salts thereof. The present invention also relates to compositions comprising a compound of the present invention and methods of treating diseases and conditions which are advantageously treated by administering an inhibitor of a Janus-associated kinase having selectivity for subtypes 1 and 2 (JAK1/JAK2). of the use of such a composition.

1 shows the results of a metabolic stability test of a referenced compound.

DETAILED DESCRIPTION OF THE INVENTION

Justice

The term "treatment" refers to reducing, inhibiting, ameliorating, curtailing, arresting, stabilizing, or alleviating the severity of a disease (eg, a disease or disorder described above) on the occurrence or progression. Or, it means to improve the symptoms associated with the disease.

The term “disease” means any condition or disorder that impairs or interferes with the standard function of a cell, tissue, or organ.

It will be appreciated that some variability in natural isotopic abundance occurs in synthesized compounds depending on the origin of the chemical substances used in the synthesis. Thus, preparations of ruxolitinib will inherently contain small amounts of deuterated isotopologues. The concentrations of stable hydrogen and carbon isotopes abundant in nature, despite this diversity, are small and non-problematic compared to the stable isotopic degrees of substitution of the compounds of the present invention. Reference Examples [Wada, E et al., Seikagaku, 1994, 66:15; Gannes, LZ et al., Comp Biochem Physiol Mol Integr Physiol, 1998, 119:725]

In the compounds of the present invention, 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 specifically designated as "H" or "hydrogen," it is understood that the position has hydrogen in its natural abundance isotopic composition. Also, unless otherwise stated, when a position is specifically designated as "D" or "deuterium," that position contains deuterium in an abundance 3000 times greater than its natural abundance of 0.015% deuterium (i.e., at least 45% deuterium incorporation). It is understood to have

As used herein, the term “isotopic abundance factor” refers to the ratio between the isotopic abundance and natural abundance of a specified isotope.

In another embodiment, the compound of the present invention is 3500 or greater (52.5% deuterium incorporation at each designated deuterium atom), 4000 or greater (60% deuterium incorporation at each designated deuterium atom), 4500 or greater (67.5% deuterium incorporation) , 5000 or more (75% deuterium), 5500 or more (82.5% deuterium mixing), 6000 or more (90% deuterium mixing), 6333.3 or more (95% deuterium mixing), 6466.7 or more (97% deuterium mixing), 6600 or more (99%) deuterium incorporation), or an isotopic abundance factor for each designated deuterium atom of 6633.3 or greater (99.5% deuterium incorporation).

The term "isotopologue" refers to a species whose chemical structure differs from a particular compound of the invention only in isotopic composition.

The term "compound" when referring to a compound of the present invention refers to a group of molecules having the same chemical structure, except that there may be isotopic variations in the constituent atoms of the molecule. Thus, it will be clear to those skilled in the art that a compound represented by a particular chemical structure containing the indicated deuterium will also contain lower amounts of isotopologues having a hydrogen atom at one or more of the designated deuterium positions in that structure. The relative amounts of such isotopologues in the compounds of the present invention will depend on many factors including the efficiency of incorporation of deuterium in the various synthetic steps used to prepare the compounds and the isotopic purity of the deuterated reagents used to prepare the compounds. will be. However, as noted above, the relative amount of all such isotopologues will be less than 49.9% of the compound. In other embodiments, the relative amount of all such isotopologues is less than 47.5%, less than 40%, less than 32.5%, less than 25%, less than 17.5%, less than 10%, less than 5%, less than 3%, less than 1% of the compound. , or less than 0.5%.

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

As used herein, the term "pharmaceutically acceptable" means, within the scope of sound medical judgment, suitable for use in contact with tissues of humans and other mammals without undue toxicity, irritation, and allergic reaction, and has a reasonable benefit/risk ratio. indicates the corresponding component. "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 the present invention. A “pharmaceutically acceptable counterion” is the ionic moiety of a salt that is not toxic when released from the salt upon administration to a recipient.

Acids commonly used to form pharmaceutically acceptable salts include inorganic acids such as hydrogen bisulfonic acid, 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-bromophenylsulf phonic acid, carbonic acid, succinic acid, citric acid, benzoic acid and acetic acid, as well as related inorganic and organic acids. Accordingly, such pharmaceutically acceptable salts 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, fuma Late, maleate, buphine-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate , terephthalate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfo nate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and other salts. In one embodiment, pharmaceutically acceptable acid addition salts include those formed with inorganic acids such as hydrochloric acid and hydrobromic acid, particularly those formed with organic acids such as maleic acid.

A compound of the present invention (eg, a compound of formula (I) or a compound of formula (A)) may contain asymmetric carbon atoms, eg, asymmetric carbon atoms as a result of deuterium substitution or otherwise. Thus, the compounds of the present invention may exist as individual enantiomers or as mixtures of two enantiomers. Accordingly, the compounds of the present invention may exist as racemic mixtures or scalemic mixtures, or as individual individual stereoisomers substantially free of other possible stereoisomers. As used herein, the term “substantially free of other stereoisomers” means less than 25% other stereoisomers, preferably less than 10% other stereoisomers, more preferably less than 5% other stereoisomers, most preferably means that less than 2% other stereoisomers are present. Methods for obtaining or synthesizing individual enantiomers for a given compound are known in the art and can be practicably applied to final compounds or to starting materials or intermediates.

Unless otherwise indicated, disclosed compounds are shown by name or structure without specifying stoichiometry and are understood to represent all possible stereoisomers of the compounds when they bear one or more centers of chiral.

As used herein, the term "mammal" refers to a human or non-human animal, such as a mouse, rat, guinea pig, dog, cat, horse, cow, pig, monkey, chimpanzee, baboon, or rhesus monkey. (rhesus). In one embodiment, the mammal is a non-human animal. In another embodiment, the mammal is a human.

The term "stable compound," as used herein, has sufficient safety to permit their manufacture and is responsive to a disease or condition for the purposes detailed herein (eg, a therapeutic product, an intermediate for use in the production of a therapeutic compound, a therapeutic agent). refers to a compound that retains the integrity of the compound for a period of time sufficient to be useful (formulation into an isolated or storable intermediate compound for the treatment of

Both "D" and "d" represent deuterium. "Stereoisomer" refers to both enantiomers and diastereomers. "tert" and "t" each represent tertiary. "US" stands for United States of America.

“Substituted with deuterium” indicates replacement of one or more hydrogen atoms with the corresponding number of deuterium atoms.

Throughout this specification, variables may be denoted generally (eg, "each R") or may be denoted specifically (eg, R ¹ , R ² , R ³ , etc.). Unless otherwise indicated, when a variable appears generally, it is meant to include all specific embodiments of that specific variable.

therapeutic compound

In one embodiment, the present invention provides a compound of formula (A) or a pharmaceutically acceptable salt thereof:

In the above formula,

Y ¹ is selected from hydrogen and deuterium;

each Y ² is independently selected from hydrogen and deuterium, provided that each Y ² bonded to a common carbon is the same;

each Y ³ is independently selected from hydrogen and deuterium, provided that each Y ³ bonded to a common carbon is the same;

Y ⁴ is selected from hydrogen and deuterium;

each Y ⁵ is the same and is selected from hydrogen and deuterium;

Y ⁶ , Y ⁷ , Y ⁸ , Y ⁹ , and Y ¹⁰ are each independently selected from hydrogen and deuterium; provided that Y ¹ is hydrogen, each Y ² and each Y ³ is hydrogen, Y ⁴ is hydrogen, and each Y ⁶ , Y ⁷ , Y ⁸ , Y ⁹ , and Y ¹⁰ is hydrogen, each of Y ₅ is deuterium.

In one embodiment of Formula A, each Y ² is the same, each Y ³ is the same, and each Y ⁵ is the same. In one aspect of this embodiment, each Y ² is deuterium. In a further aspect, each Y ³ is deuterium. In yet a further aspect, each Y ³ is hydrogen. In another aspect of this embodiment, each Y ² is hydrogen. In a further aspect, each Y ³ is deuterium. In yet a further aspect, each Y ³ is hydrogen. In one example of any of the aforementioned aspects, Y ¹ is deuterium. In another of any of the preceding aspects, Y ¹ is hydrogen. In further specific examples of any of the preceding embodiments, Y ¹ is deuterium, Y ⁴ is deuterium, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is deuterium, Y ⁴ is deuterium, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is deuterium, Y ⁴ is hydrogen, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is hydrogen, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is hydrogen, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is deuterium, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is deuterium, and each Y ⁵ is hydrogen.

In one embodiment, Y ⁶ is deuterium. In one aspect of this embodiment, each of Y ⁷ and Y ⁸ is deuterium. In another aspect of this embodiment, each of Y ⁷ and Y ⁸ is hydrogen.

In one embodiment, Y ⁶ is hydrogen. In one aspect of this embodiment, each of Y ⁷ and Y ⁸ is deuterium. In another aspect of this embodiment, each of Y ⁷ and Y ⁸ is hydrogen.

In one embodiment, the present invention provides a compound of formula (I): or a pharmaceutically acceptable salt thereof:

In the above formula,

Y ¹ is selected from hydrogen and deuterium;

each Y ² is independently selected from hydrogen and deuterium, provided that each Y ² bonded to a common carbon is the same;

each Y ³ is independently selected from hydrogen and deuterium, provided that each Y ³ bonded to a common carbon is the same;

Y ⁴ is selected from hydrogen and deuterium;

each Y ⁵ is the same and is selected from hydrogen and deuterium;

Y ⁶ , Y ⁷ , and Y ⁸ are each independently selected from hydrogen and deuterium;

with the proviso that each Y ⁵ is deuterium ^{when Y 1} is hydrogen, each Y ² and each Y ³ is hydrogen, Y ⁴ is hydrogen, and each Y ⁶ , Y ⁷ and Y ^{8 is hydrogen.}

In one embodiment, each Y ² is the same, each Y ³ is the same, and each Y ⁵ is the same. In one aspect of this embodiment, each Y ² is deuterium. In a further aspect, each Y ³ is deuterium. In yet a further aspect, each Y ³ is hydrogen. In another aspect of this embodiment, each Y ² is hydrogen. In a further aspect, each Y ³ is deuterium. In yet a further aspect, each Y ³ is hydrogen. In one example of any of the aforementioned aspects, Y ¹ is deuterium. In another example of any of the preceding aspects, Y ¹ is hydrogen. In further specific examples of any of the preceding embodiments, Y ¹ is deuterium, Y ⁴ is deuterium, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is deuterium, Y ⁴ is deuterium, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is deuterium, Y ⁴ is hydrogen, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is hydrogen, and each Y ⁵ is hydrogen. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is hydrogen, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is deuterium, and each Y ⁵ is deuterium. In still further specific examples of any of the above aspects, Y ¹ is hydrogen, Y ⁴ is deuterium, and each Y ⁵ is hydrogen.

In one embodiment, Y ⁶ is deuterium. In one aspect of this embodiment, each of Y ⁷ and Y ⁸ is deuterium. In another aspect of this embodiment, each of Y ⁷ and Y ⁸ is hydrogen.

In one embodiment, Y ⁶ is hydrogen. In one aspect of this embodiment, each of Y ⁷ and Y ⁸ is deuterium. In another aspect of this embodiment, each of Y ⁷ and Y ⁸ is hydrogen.

In one embodiment, the compound is a compound of Formula I, wherein ^{Y 6 , Y 7} and Y ⁸ are each hydrogen, the compound being any of the compounds (Cmpd) listed in Table 1 (below), or a pharmaceutically acceptable compound thereof. salts, wherein any atom not denoted as deuterium is present in its natural isotopic abundance.

Table 1: Exemplary embodiments of Formula I

In one embodiment, the compound is a compound of Formula I, wherein ^{Y 6 , Y 7} and Y ⁸ are each D, the compound being any of the compounds (Cmpd) listed in Table 2 (below) or a pharmaceutically acceptable compound thereof. salts, wherein any atom not denoted as deuterium is present in its natural isotopic abundance.

Table 2: Exemplary embodiments of Formula I

In another set of embodiments, any atom not designated as deuterium in any of the aforementioned embodiments is present in its natural isotopic abundance.

The following compounds, or salts thereof, are useful for preparing several compounds of the present invention, wherein any atom not designated as deuterium is present in its natural isotopic abundance:

The synthesis of a compound of Formula I or Formula A can be readily accomplished by a synthetic chemist of ordinary skill with reference to the exemplary syntheses and examples described herein. Related procedures analogous to those used to prepare compounds of Formula I or Formula A and intermediates thereof are described, for example, in US Pat. No. 7,598,257 and Organic Letters, 2009, 11(9): 1999-2009. have.

These methods employ the corresponding deuterated and optionally other isotopically containing reagents and/or intermediates to synthesize the compounds described herein, or apply standard known synthetic protocols for introducing isotopic atoms into chemical structures. can be performed.

Exemplary Synthesis

Compounds of formula (I) or formula (A) can be prepared in a manner analogous to those syntheses set forth in U.S. Pat. No. 7,598,257 and Organic Letters, 2009, 11(9): 1999-2009 using suitably deuterated starting materials. can

Compounds of formula (I) or formula (A) can also be prepared as shown in the schemes below.

Scheme 1. Preparation of compounds of formula (I).

Scheme 1 discloses an exemplary preparation of a compound of Formula I, wherein Y ¹ , each Y ² and each Y ³ is deuterium, Y ⁴ , each Y ⁵ , Y ⁶ , Y ⁷ and Y ⁸ is hydrogen. In a manner analogous to that described in WO 2010/083283, commercially available 4-chloro-7H-pyrrolo[2,3-d]pyrimidine 11 (Aldrich) was treated with sodium hydride and SEM chloride to obtain 12 provided, which reacts with commercially available 13 to give 14. 11 instead of 4-Bromo-7H-pyrrolo[2,3-d]pyrimidine was also used in the first step to SEM-protected 4-bromo-7H-pyrrolo[2,3-d]pyrimidine ( 12 and it may provide similar), which can provide a 14 to react with 13. The reactions of 15 and 14 prepared as described in Scheme 2a below are described in Lin, Q. et al. Org. Lett. 2009, 11, 1999] to give 16 . The reaction is carried out in the presence of a chiral ligand 27 , prepared as described in Lin, Q. et al. 16 is converted to 17 by treatment with _{NH 4} OH and I _{2 .} Thereafter, the SEM protecting group of 17 _{is deprotected with LiBF 4} and NH ₄ OH to give the compound of formula (I).

Scheme 2a . Preparation of compound 15.

As shown in scheme 2a, 18 commercially available is processed into a phosphonium one lead 20 and DCl / D ₂ O provides 19 and 19 is processed to 20 and DiBAl-H provides 15.

Scheme 2b . Preparation of compound 23.

Scheme 2c . Preparation of compound 26.

Compounds similar to 15 can also be prepared. For example, as shown in Scheme 2b, commercially available 21 can be converted to 23 in a manner similar to that disclosed in Scheme 2a. As another example, as shown in Scheme 2c, commercially available 24 can be converted to 26 in a manner similar to that disclosed in Schemes 2a and 2b. 23 can be converted to a compound of formula I in a manner analogous to that disclosed in Scheme 1, wherein Y ¹ and each Y ³ are deuterium, Y ⁴ , each Y ² , each Y ⁵ , Y ⁶ , Y ⁷ and Y ⁸ are hydrogen. Likewise, 26 can be converted to a compound of formula I in a manner analogous to that disclosed in Scheme 1, wherein Y ¹ and each Y ² are deuterium, Y ⁴ , each Y ³ , each Y ⁵ , Y ⁶ , Y ⁷ and Y ⁸ are hydrogen.

The specific methods and compounds shown above are not intended to be limiting. Chemical structural formulas in the schemes herein ^{are identified or not identified by the same variable name (ie, R 1} , R ² , R ^{3 ,} etc.) chemical group definitions (moieties, atoms, etc.) at positions corresponding to the formulas of the compounds herein. Variables defined herein as suitable for The suitability of a chemical group in a compound structural formula used in the synthesis of another compound is within the knowledge of those skilled in the art.

Additional methods of synthesizing compounds of Formula I or Formula A and their synthetic precursors, including those within routes not explicitly shown in the schemes herein, are within the means of a chemist skilled in the art. In synthesizing applicable compounds useful synthetic chemical change and protecting group methodologies (protection and deprotection) are known in the art, see, e.g., [Larock R, Comprehensive Organic Transformations , VCH Publishers (1989); Greene, TW et al., Protective Groups in Organic Synthesis , 3 ^rd Ed., John Wiley and Sons (1999); Fieser, L et al., Fieser and Fieser's Reagents for Organic Synthesis , John Wiley and Sons (1994); and Paquette, L, ed., Encyclopedia of Reagents for Organic Synthesis , John Wiley and Sons (1995) and their subsequent editions.

Combinations of substituents and variables contemplated by the present invention are only those that result in the formation of stable compounds.

composition

The present invention also provides an effective amount of a compound of Formula I or Formula A (eg, including any of the formulas herein), or a pharmaceutically acceptable salt of said compound; and a pharmaceutically acceptable carrier. A carrier(s) is "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof in the amounts used in the medicament.

Pharmaceutically acceptable carriers, adjuvants and excipients that can be used in the pharmaceutical compositions of the present invention include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphate, 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 salt, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat, It is not limited thereto.

If desired, the solubility and bioavailability of a compound of the present invention in a pharmaceutical composition can be improved by methods well known in the art. One method involves the use of lipid excipients in the formulation [see: "Oral Lipid-Based Formulations: Enhancing the Bioavailability of Poorly Water-Soluble Drugs (Drugs and the Pharmaceutical Sciences)," David J. Hauss, ed. Informa Healthcare, 2007; and "Role of Lipid Excipients in Modifying Oral and Parenteral Drug Delivery: Basic Principles and Biological Examples," Kishor M. Wasan, ed. Wiley-Interscience, 2006].

Another known method of enhancing bioavailability is the use of a compound of the invention in an amorphous form optionally formulated with a poloxamer, such as LUTROL ^™ and PLURONIC ^™ (BASF Corporation), or a block copolymer of ethylene oxide and propylene oxide. See U.S. Patent Nos. 7,014,866; and US Patent Publication Nos. 20060094744 and 20060079502].

Pharmaceutical compositions of the present invention include those suitable for oral, rectal, intranasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous and intradermal) administration. In certain embodiments, a compound of a formula herein is administered transdermally (eg, using a transdermal patch or iontophoretic technique). Other dosage forms may conveniently be presented in unit dosage form, such as tablets, sustained release capsules, and liposomes, and may be prepared by any method well known in the art of pharmacy. See Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, Baltimore, MD (20th ed. 2000)].

Such preparations include the step of combining an ingredient, such as a carrier, which constitutes one or more accessory ingredients, with the molecule to be administered. In general, the compositions are prepared uniformly and intimately and combined with the active ingredient with liquid carriers, liposomes or finely divided solid carriers, or both, and, if necessary, molded into an article.

In certain embodiments, the compound is administered orally. Compositions of the present invention suitable for oral administration each comprise a predetermined amount of the active ingredient; powder or granules; solutions or suspensions in aqueous or non-aqueous liquids; oil-in-water liquid emulsions; It may contain an aqueous-in-oil liquid emulsion and may be presented as discrete units, such as capsules, sachets, or tablets, packed with liposomes, or as a bolus or the like. Soft gelatin capsules may be useful for containing such suspensions, which may advantageously increase the rate of compound absorption.

In the case of tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also typically added. For oral administration in capsule form, useful diluents include lactose and dry corn starch. 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.

Compositions suitable for oral administration include lozenges comprising flavor-based ingredients, usually sucrose and acacia or tragacanth; and pastilles comprising active ingredients on an inert basis, such as gelatin and glycerin, or sucrose and acacia.

Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injectable solutions which may contain antioxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may contain suspending and thickening agents. The formulations may be presented in unit-dose or multi-dose containers, for example, hermetically sealed ampoules and vials, and stored in a freeze-dried (lyophilized) state immediately prior to use in a sterile liquid carrier, e.g., water for injection. Only additions may be required. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

Such injectable solutions may be in the form of, for example, sterile injectable aqueous or oleaginous suspensions. Such suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents (eg, such as Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable excipients and solvents that may be used are mannitol, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as the solvent or suspending medium. For this purpose, any bland fixed oil may be used, including synthetic mono- or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful for the preparation of injectable natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially polyoxyethylated versions thereof. Such oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants.

The pharmaceutical composition of the present invention may be administered in the form of a suppository for rectal administration. Such compositions can be prepared by mixing a compound of the present invention with a suitable non-irritating excipient, which is solid at room temperature but liquid at rectal temperature and will melt in the rectum to release the active ingredient. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycol.

The pharmaceutical composition of the present invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation, benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons, and/or other stabilizing or dispersing agents known in the art. (see, e.g., Rabinowitz JD and Zaffaroni AC, US Pat. No. 6,803,031 assigned to Alexza Molecular Delivery Corporation).

Topical administration of the pharmaceutical compositions of the present invention is particularly useful when the desired treatment involves areas or organs readily accessible by topical application. For topical topical application to the skin, the pharmaceutical composition should be formulated in a suitable ointment containing the active ingredient suspended or dissolved in a carrier. Carriers for topical administration of a compound of the present invention include, but are not limited to, mineral oil, liquefied petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax, and water. Alternatively, the pharmaceutical composition may be formulated into a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol, and water. The pharmaceutical compositions of the present invention may also be applied topically to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topical-transdermal patches and iontophoretic administration are also encompassed by the present invention.

Application of the therapeutics of the present invention may be topical to be administered to the site of interest. Various techniques, such as injections, use of catheters, trocars, projectiles, pluronic gels, stents, sustained release drug release polymers or other devices that allow in vivo access can be used to apply the composition of the present invention to the region of interest.

Thus, according to yet another embodiment, the compounds of the present invention may be included in compositions for coating implantable medical devices, such as prostheses, artificial valves, vascular grafts, stents, or catheters. Suitable coatings and general preparation of coated implantable devices are known in the art and are described in U.S. Patent Nos. 6,099,562; 5,886,026; and 5,304,121. The coating is typically a biocompatible polymeric material such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coating may optionally be further covered with a suitable topcoat of a fluorosilicone, polysaccharide, polyethylene glycol, phospholipid or combination thereof to impart controlled release characteristics in the composition. Coatings for invasive devices should be included within the definition of the term pharmaceutically acceptable carrier, adjuvant or excipient as used herein.

According to another embodiment, the present invention provides a method of coating an implantable medical device comprising the step of contacting the implantable medical device with the coating composition described above. It will be apparent to those skilled in the art that the coating of the device will be made prior to implantation into a mammal.

According to another embodiment, the present invention provides a method of impregnating an implantable drug release device comprising the step of contacting the implantable drug release device with a compound or composition of the present invention.

Implantable drug release devices include, but are not limited to, biodegradable polymer capsules or bullets, non-degradable, diffusible polymer capsules, and biodegradable polymer wafers.

According to another embodiment, the present invention provides an implantable medical device coated with a compound or a composition comprising a compound of the present invention, wherein the compound is therapeutically active.

According to another embodiment, the present invention provides an implantable drug release device into which a compound or a composition comprising a compound of the present invention is injected, wherein the compound is released from the device and is therapeutically active.

When an organ or tissue becomes available upon removal from the subject, the organ or tissue may be immersed in a medium containing the composition of the present invention, or the composition of the present invention may be applied to the organ, or the present invention may The composition may be applied in any other conventional manner.

In another embodiment, the composition of the present invention further comprises a second therapeutic agent. The second therapeutic agent may be selected from any compound or therapeutic agent known to have or exhibit beneficial properties when administered with a compound having the same mechanism of action as ruxoritinib. Such agents include those described as useful in combination with ruxolitinib.

Preferably, the second therapeutic agent is primary myelofibrosis, polycythemia vera, post-polycythemia vera myelofibrosis, chronic idiopathic myelofibrosis, the present invention Myelofibrosis, including post-essential thrombocythemia myelofibrosis, and essential thrombocythemia, pancreatic cancer, prostate cancer, breast cancer, leukemia (leukemia), non-Hodgkin's lymphoma, multiple myeloma, psoriasis and alopecia areata are useful agents for the treatment or prevention of a disease or condition selected from.

In one embodiment, the second therapeutic agent is selected from lenalidomide, panobinostat, capecitabine, exemestane, and combinations thereof.

In another embodiment, the invention provides separate dosage forms of a compound of the invention and one or more of any of the second therapeutic agents described above, wherein the compound and the second therapeutic agent are in association with each other. As used herein, the term “associated with each other” means that the separate dosage forms are packaged together or otherwise attached to each other, whereby the separate dosage forms are purchased and administered together (within less than 24 hours of each other, sequentially or simultaneously). ) is readily apparent.

In the pharmaceutical compositions of the present invention, the compound of the present invention is present in an effective amount. As used herein, the term “effective amount” refers to an amount sufficient to treat a target disease when administered in an appropriate dosing regimen.

The correlation of dose (based on milligrams per square meter of body surface) for animals and humans is described in Freireich et al., Cancer Chemother. Rep, 1966, 50: 219. The body surface area can be approximately determined from the height and weight of the subject. See, eg, Scientific Tables, Geigy Pharmaceuticals, Ardsley, N.Y., 1970, 537.

In one embodiment, an effective amount of a compound of the present invention may range from 1 mg to 500 mg, such as 5 mg to 100 mg, such as 5 mg to 50 mg. Examples of ranges are 40 mg to 50 mg, 25 mg to 40 mg, 25 mg to 50 mg, 20 mg to 40 mg, 20 mg to 50 mg, 10 mg to 25 mg, 10 mg to 20 mg, 5 mg to 25 mg, 5 mg to 20 mg, and 5 mg to 10 mg. In one embodiment, doses of 10 mg, 20 mg, 40 mg, and 50 mg are administered once daily. In one embodiment, doses of 5 mg, 10 mg, 20 mg, 40 mg, and 50 mg are administered twice daily.

An effective dose also depends on the disease being treated, the severity of the disease, the route of administration, the subject's sex, age and general health, excipient usage, concomitant use with other therapeutic treatments, e.g., other This will vary depending on the availability of the agent and the judgment of the treating physician. For example, guidelines for selecting an effective dose may be determined with reference to prescribing information for ruxoritinib.

For example, for a pharmaceutical composition comprising a second therapeutic agent, an effective amount of the second therapeutic agent is from about 20% to 100% of the dosage typically used in monotherapy regimens employing the agent. Preferably, the effective amount is from about 70% to 100% of a typical monotherapeutic dose. Typical monotherapeutic dosages of the second therapeutic agent are well known in the art. See, for example, Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000)].

It is expected that some of the aforementioned second therapeutic agents will act synergistically with the compounds of the present invention. If synergy occurs, the effective dosage of the second therapeutic agent and/or the compound of the present invention will be reduced from the effective dosage required in monotherapy. This has the advantage of minimizing the toxic side effects of the second therapeutic agent of the compound of the invention, a synergistic improvement in efficacy, improved ease of administration or use, and/or reduced overall consumption of the compound preparation or formulation.

treatment method

In another embodiment, the present invention provides Janus Associated Kinase (JAK) JAK1 and JAK2 in a cell comprising contacting the cell with one or more compounds of Formula I or Formula A herein, or a pharmaceutically acceptable salt thereof A method of inhibiting one or more of

According to another embodiment, the present invention provides ruxortinib in a subject in need of treatment of a disease advantageously treated by ruxoritinib comprising administering to the subject an effective amount of a compound or composition of the present invention. It provides a method of treating a disease that is advantageously treated by In one embodiment, the subject is a patient in need of such treatment. Such diseases are well known in the art and are disclosed in, but not limited to, US Pat. No. 7,598,257. The disease may include diseases associated with the immune system, such as organ transplant rejection (eg, allograft rejection and graft-versus-host disease); autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, juvenile arthritis, type I diabetes, lupus, psoriasis, inflammatory bowel disease, ulcerative colitis, Crohn's disease, myasthenia gravis, immunoglobulin nephropathy, autoimmune thyroid disease; allergic diseases such as asthma, food allergy, atopic dermatitis and rhinitis; viral diseases such as Epstein Barr virus (EBV), hepatitis B, hepatitis C, HIV, HTLV1, varicella-zoster virus (VZV) and human papillomavirus (HPV); skin diseases such as psoriasis (eg, psoriasis vulgaris), atopic dermatitis, skin rash, skin irritation, skin hypersensitivity (eg, contact dermatitis or allergic contact dermatitis); Cancer, eg, a cancer characterized by a solid cancer (eg, prostate cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, Kaposi's sarcoma, Castleman's disease, melanoma) tumors), hematological cancers (e.g., lymphoma, leukemia, e.g., acute lymphoblastic leukemia, or multiple myeloma), and skin cancers, e.g., cutaneous T-cell lymphoma (CTCL) and cutaneous B-cell lymphoma ( examples thereof include Sézary syndrome and mycosis fungoides); Myeloproliferative diseases (MPDs) such as polycythemia vera (PV), essential thrombocythemia (ET), myeloid metaplasia with myelofibrosis (MMM), chronic myelogenous monocytic leukemia (CMML), hypereosinophilic syndrome (HES) ), systemic mast cell disease (SMCD); Inflammatory and inflammatory diseases, such as inflammatory diseases of the eye (eg, iritis, uveitis, scleritis, conjunctivitis, or related diseases), inflammatory diseases of the airways (eg, upper respiratory tract, including nose and sinuses) eg rhinitis or sinusitis or lower respiratory tract, eg bronchitis, chronic obstructive pulmonary disease, etc.), inflammatory myopathies, eg myocarditis; systemic inflammatory response syndrome (SIRS) and septic shock; diseases or disorders associated with ischemic reperfusion injury or inflammatory ischemic events, eg, stroke or heart attack; loss of appetite; cachexy; fatigue, such as arising from or associated with cancer; restenosis; sclerodermitis; fibrosis; diseases associated with hypoxia or astrogliosis, such as diabetic retinopathy, cancer, or neurodegeneration; ventilation; Examples include, but are not limited to, benign prostatic hyperplasia or increased prostate size due to benign prostatic hyperplasia.

In one specific embodiment, the method of the present invention is for treating myelofibrosis, e.g., primary myelofibrosis, myelofibrosis after polycythemia vera, myelofibrosis after essential thrombocytosis, essential thrombocytopenia, or a combination thereof; pancreatic cancer; prostate cancer; breast cancer; Leukemia, Non-Hodgkin's Lymphoma; multiple myeloma; for treating a disease or condition selected from psoriasis and combinations thereof in a subject in need thereof.

In another specific embodiment, the method of the present invention is used in a subject in need of treatment for a disease or condition selected from myelofibrosis, e.g., primary myelofibrosis, myelofibrosis after polycythemia vera and myelofibrosis after essential thrombocytopenia. used to treat the disease or condition.

Identification of a subject in need of such treatment may be at the discretion of the subject or health care professional, which may be subjective (eg, opinion) or objective (eg, measurable by a test or diagnostic method). ) can be

In another embodiment, any of the above methods of treatment comprise the additional step of co-administering one or more second therapeutic agents to a subject in need thereof. The choice of second therapeutic agent can be from any second therapeutic agent known to be useful for co-administration with ruxoritinib. The choice of the second therapeutic agent also depends on the particular disease or condition being treated. Examples of second therapeutic agents that can be used in the methods of the invention are those described above for use in combination compositions comprising a compound of the invention and a second therapeutic agent.

In particular, the combination therapy of the present invention comprises the co-administration of a compound of formula (I) or formula (A) and a second therapeutic agent to a subject in need thereof for the treatment of the following diseases (the specific second therapeutic agent indicated in parentheses after the indications) using): myelofibrosis (lenalidomide or panobinostat); pancreatic cancer (capecitabine); and breast cancer (exemestane).

As used herein, the term “co-administered” means that the second therapeutic agent is part of a single dosage form (eg, a composition of the present invention comprising a compound of the present invention and a second therapeutic agent as described above) or separate or separate. It means that it can be administered with the compound of the present invention in multiple dosage forms. Alternatively, the additional agent may be administered prior to, sequentially with, or after administration of the compound of the present invention. In such combination therapy treatment, both the compound of the invention and the second therapeutic agent(s) are administered by conventional methods. Administration of a composition of the present invention comprising both a compound of the present invention and a second therapeutic agent to a subject may result in said same therapeutic agent, any other second therapeutic agent, or any compound of the present invention at another time during the course of treatment. Separate administration to the subject is not excluded.

Effective amounts of such second therapeutic agents are well known to those skilled in the art, and instructions for administration are provided in the patents and published patent applications referred to herein, as well as in Wells et al., eds., Pharmacotherapy Handbook, 2nd Edition, Appleton and Lange, Stamford, Conn. (2000); PDR Pharmacopoeia, Tarascon Pocket Pharmacopoeia 2000, Deluxe Edition, Tarascon Publishing, Loma Linda, Calif. (2000)], and other medical textbooks. However, it is completely within the skill of one of ordinary skill in the art to determine the optimal effective amount range of the second therapeutic agent.

In one embodiment of the invention, when a second therapeutic agent is administered to a subject, the effective amount of a compound of the invention is less than its effective amount that would be present if the second therapeutic agent was not administered. In another embodiment, the effective amount of the second therapeutic agent is less than its effective amount that would be present if the compound of the present invention was not administered. In this way, the undesirable side effects associated with high doses of either agent can be minimized. Other potential advantages (including, but not limited to, improved dosing regimens and/or reduced drug costs) will be apparent to those skilled in the art.

In another embodiment, the present invention provides a compound of Formula I or Formula A alone or an agent described above in the manufacture of a medicament as a single composition or separate dosage form for the treatment or prophylaxis in a subject of a disease, disorder or condition described above. 2 provides for use with one or more of the therapeutic agents. Another aspect of the present invention is a compound of formula (I) or formula (A) for use in the treatment or prophylaxis in a subject of a disease, condition or symptom thereof described herein.

Example

Example One . ( R )-3-(4-(7H- pyrrolo[2,3-d]pyrimidine -4-yl)-1H- pyrazole -1-yl)-3-(2,2,5,5- d ₄ -cyclopentyl) propanenitrile (compound 107 ) synthesis.

Scheme 3. Compound 1 07 manufacture of

Step 1. Diethyl 2,2,5,5- d ₄ - cyclopentane-1,1-dicarboxylate 32. To a solution of diethyl malonate (6.57 mL, 43.3 mmol) in ethanol (40 mL) was added a 21 wt % solution of sodium ethoxide in ethanol (32.3 mL, 86.6 mmol) followed by 1,1,4,4- Tetradeutero-1,4-dibromobutane ( 31 , 5.53 mL, 45.5 mmol, CDN isotope, 98 atomic % D) was added. The resulting solution was stirred at reflux for 2 h, then cooled to room temperature and diluted with excess water. Then, most of the ethanol was removed through distillation, and the resulting aqueous solution was extracted with ethyl acetate (3 x 75 mL). The organic layers were combined, washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give 32 as a yellow oil, which was used in the next step without purification (9.45 g, 100). %).

Step 2. 2,2,5,5- d ₄ - cyclopentane-1-carboxylic acid (33). To a solution of compound 32 (9.45 g, 43.3 mmol) in ethanol (20 mL) was added a 5M solution of sodium hydroxide (20 mL). Additional water (15 mL) was then added and the reaction stirred at reflux for 3 h. When cooled to room temperature, the reaction was diluted with excess water and most of the ethanol was removed via distillation. The aqueous solution was made acidic (pH<2) with 1N HCl and subsequently extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting bright orange solid was transferred to a pressure flask and water (140 mL) was added. The pressure flask was sealed and the reaction stirred at 160° C. for 15 h before cooling to room temperature. The reaction was diluted with 1N HCl and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 33 (4.37 g, 86%) as an amber oil, which was used without purification.

Step 3. 2,2,5,5- d ₄ -N -methoxy - N-methylcyclopentanecarboxamide ( 34 ) . In a solution of compound 33 (4.37 g, 37.0 mmol) in acetonitrile (60 mL) at 0° C. N,O-dimethylhydroxylamine hydrochloride (4.33 g, 44.4 mmol), TBTU (12.5 g, 38.9 mmol) and N ,N-diisopropylethylamine (19.0 mL, 111 mmol) was added. The reaction was stirred at room temperature for 15 h, then diluted with 1N HCl and extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with saturated NaHCO ₃ , dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The obtained product was purified by column chromatography (SiO ₂ , 0-50% ethyl acetate/hexanes) to give compound 34 (2.22 g, 37%) as a clear oil. MS (ESI) 162.3 [(M + H) ⁺ ].

Step 4. 2,2,5,5- d ₄ - cyclopentane-1-carboxaldehyde 35. To a solution of compound 34 (2.22 g, 13.8 mmol) in _{THF (50 mL) at 0° C. was added dropwise a 1M solution of LiAlH 4} in THF (24.8 mL, 24.8 mmol). The reaction was stirred at 0° C. for 1 h before it was quenched by sequential dropwise addition of water (940 μL), 15% NaOH (940 μL) and water (2.82 mL). The quenched reaction stirred at room temperature for 30 min was then filtered through Celite® and concentrated under reduced pressure. The resulting oil was diluted with 1N HCl and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 35 (850 mg, 60%) as a clear oil, which was used without purification.

Step 5. 3- (2,2,5,5- d ₄ - cyclopentyl) acrylonitrile (36). To a 1M solution of potassium tert-butoxide in THF (8.74 mL, 8.74 mmol) was added dropwise a solution of diethyl cyanomethylphosphonate (1.48 mL, 9.15 mmol) in THF (12 mL) at 0°C. The reaction was allowed to warm to room temperature, stirred for 15 minutes and then cooled to 0°C. Then aldehyde 35 (850 mg, 8.32 mmol) was added dropwise as a solution in THF (3 mL). The reaction was stirred at room temperature for 48 h, then diluted with excess water and extracted with diethyl ether (1 x 50 mL) and ethyl acetate (3 x 50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 36 (1.17 g, >100%) as a light orange oil, which was used without purification.

Step 6. (+/-) - (4- (1- (2-cyano -1- (2,2,5,5- d ₄ - cyclopentyl) ethyl) -1H- pyrazol-4-yl) -7H -pyrrolo[2,3-d]pyrimidin- 7-yl) methyl Pivalate ( (+/-)38 ) . To a solution of compound 37 (400 mg, 1.34 mmol, the preparation described in Lin, Q. et al. Org. Lett., 2009, 11, 1999-2002) in acetonitrile (10 mL) was added compound 36 (418 mg) , 3.34 mmol) was added followed by DBU (421 μL, 2.81 mmol). After that, the reaction mixture was stirred at room temperature for 15 hours and concentrated under vacuum under reduced pressure. The resulting crude mixture was diluted with water and extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with 1N HCl, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. Compound (+/- ) purified via normal phase column chromatography (SiO ₂ , 0-60% ethyl acetate/hexanes) followed by reverse phase column chromatography (C18, water containing 5-70% acetonitrile/0.1% formic acid) ) 38 (68 mg, 12%) was obtained as a white foam.

Step 7. (R)-(4-(1-(2- cyano- 1-(2,2,5,5 - tetradeuterocyclopentyl)ethyl)-1H-pyrazol-4-yl)-7H -pyrrolo [2,3-d] pyrimidin-7-yl) methyl Pivalate ( (R)-38 ) . Racemic (+/-)38 (62 mg) was dissolved in acetonitrile to a concentration of 30 mg/mL and a Daicel ChiralPak AD column (20 x 250 ) using 500 μL of compound (+/-)38 solution per injection. mm, 10 μm) by preparative HPLC using isocratic method: 30% isopropanol (+ 0.1% diethylamine)/70% hexanes (+ 0.1% diethyl) at a flow rate of 17 mL/min. amines). Baseline separations were achieved under these conditions with compound (S)-38 eluting at 15.0 min and compound (R)-38 eluting at 20.2 min.

Fractions containing each enantiomer were pooled and concentrated to give 28 mg of compound (S)-38 as a colorless film and 29 mg of compound (R)-38 as a colorless film.

Step 8. (R)-3-(4-(7H -pyrrolo[2,3-d]pyrimidin- 4-yl)-1H- pyrazol -1-yl)-3-(2,2,5 , between the claw-pentyl 5-te Tra dew Tero) propanenitrile (compound 107). Compound (R)-38 (28 mg, 0.066 mmol, 1 eq) was dissolved in methanol (1 mL) in a 20 mL scintillation vial. Sodium hydroxide (0.13 mL of a 1M solution, 0.13 mmol, 2 eq) was added and the reaction was stirred at room temperature for 18 h. The reaction was diluted with water (10 mL) and brine (20 mL). The aqueous mixture was extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered and evaporated. The crude material was purified using an Analogix automated chromatography system eluting with 0-6% methanol in dichloromethane. Product fractions were pooled and evaporated to give compound 107 as a white foam. Chiral purity >99% ee (Chiralpak OD 4.6 x 250 mm, 10 um, 70% (hexane + 0.1% diethylamine) + 30% (isopropanol + 0.1% diethylamine), 1 mL/min, 254 nm residence time = 8.85 min).

Example 2. (R)-3-(4-(7H -pyrrolo[2,3-d]pyrimidin- 4-yl)-1H- pyrazol -1-yl)-3-(3,3,4,4- d Synthesis of ₄ -cyclopentyl) propanenitrile (Compound 103 ).

Scheme 4. Compounds 103 manufacture of

Step 1. diethyl 3,3,4,4- d ₄ - cyclopentane-1,1-dicarboxylate 40. To a solution of diethyl malonate (3.25 mL, 21.4 mmol) in ethanol (20 mL) a 21 wt % solution of sodium ethoxide in ethanol (16.0 mL, 42.8 mmol) followed by 2,2,3,3-tetradeutero -1,4-dibromobutane ( 39 , 4.95 g, 22.5 mmol, CDN isotope, 98 atomic %D) was added. The resulting solution was stirred at reflux for 2 h, then cooled to room temperature and diluted with excess water. After that, most of the ethanol was removed through distillation, and the resulting aqueous solution was extracted with ethyl acetate (3 x 75 mL). The organic layers were combined, washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 40 as a yellow oil, which was used in the next step without purification. (4.67 g, 100%).

Step 2. 3,3,4,4- d ₄ - cyclopentane-1-carboxylic acid (41). To a solution of compound 40 (4.67 g, 21.4 mmol) in ethanol (10 mL) was added a 5M solution of sodium hydroxide (10 mL). Additional water (10 mL) was then added and the reaction stirred at reflux for 3 h. When cooled to room temperature, the reaction was diluted with excess water and most of the ethanol was removed via distillation. The aqueous solution was made acidic (pH<2) with 1N HCl and subsequently extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting bright orange solid was transferred to a pressure flask and water (70 mL) was added. The pressure flask was sealed and the reaction stirred at 160° C. for 15 h before cooling to room temperature. The reaction was diluted with 1N HCl and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 41 (1.93 g, 76%) as an amber oil, which was used without purification.

Step 3. 3,3,4,4- d ₄ -N -methoxy - N-methylcyclopentanecarboxamide ( 42 ) . To a solution of compound 41 (1.93 g, 16.3 mmol) in acetonitrile (30 mL) at 0° C. N,O-dimethylhydroxylamine hydrochloride (1.91 g, 19.6 mmol), TBTU ((5.50 g, 17.1 mmol) and N,N-diisopropylethylamine (8.52mL, 48.9mmol) was added.The reaction was stirred at room temperature for 15 hours, then diluted with 1N HCl and extracted with ethyl acetate (3×50mL).The organic layers were combined and , washed with saturated NaHCO ₃ , dried (Na ₂ SO ₄ ), filtered, and concentrated under reduced pressure The resulting product was purified by column chromatography (SiO ₂ , 0-40% acetone/hexane) to compound 42 (1.47 g, 56%)) was obtained as a clear oil. MS (ESI) 162.3 [(M + H) ⁺ ].

Step 4. 3,3,4,4- d ₄ - cyclopentane-1-carboxaldehyde 43. To a solution of compound 42 (1.47 g, 9.12 mmol) in _{THF (35 mL) at 0° C. was added a 1M solution of LiAlH 4} in THF (16.4 mL, 16.4 mmol) dropwise. The reaction was then stirred at room temperature for 1 h at 0° C. and quenched by sequential dropwise addition of water (623 μL), 15% NaOH (623 μL) and water (1.87 mL). The quenched reaction stirred at room temperature for 30 min was then filtered through Celite® and concentrated under reduced pressure. The resulting oil was diluted with 1N HCl and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 43 (767 mg, 82%) as a clear oil, which was used without purification.

Step 5. 3- (3,3,4,4- d ₄ - cyclopentyl) acrylonitrile (44). To a solution of diethyl cyanomethylphosphonate (0.607 mL, 3.75 mmol) in THF (10 mL) at 0° C. was added dropwise a 1M solution of potassium tert-butoxide in THF (3.75 mL, 3.75 mmol). The reaction was stirred at 0° C. for 1 hour. Then aldehyde 43 (767 mg, 7.51 mmol) was added dropwise as a solution in THF (3 mL). The reaction was stirred at room temperature for 15 h, then diluted with an excess of 1:1 water/brine and extracted with MTBE (3 x 50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting oil _{was dissolved in CH 2} Cl ₂ (100 ml) and washed with NaHSO ₃ (3×25 mL). The organic layer was dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 44 (537 mg, 57%) as a light orange oil, which was used without purification.

Step 6. (+/-) - (4- (1- (2-cyano -1- (3,3,4,4- d ₄ - cyclopentyl) ethyl) -1H- pyrazol-4-yl) -7H -pyrrolo[2,3-d]pyrimidin- 7-yl) methyl Pivalate ( (+/-)45 ) . In a solution of compound 37 (514 mg, 1.72 mmol, the preparation described by Lin, Q. et al. Org. Lett., 2009, 11, 1999-2002) in acetonitrile (15 mL) , compound 44 (537 mg) , 4.29 mmol) was added followed by DBU (540 μL, 3.61 mmol). The reaction was stirred at room temperature for 15 hours and then concentrated under vacuum under reduced pressure. The resulting crude mixture was diluted with water and extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with 1N HCl, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. Purification via normal phase column chromatography (SiO ₂ , 0-60% ethyl acetate/hexanes) gave compound (+/-)45 (368 mg, 50%) as a white foam.

Step 7. (R) - (4- ( 1- (2- cyano -1- (3,3,4,4- d ₄ - cyclopentyl) ethyl) -1H- pyrazol-4-yl) -7H -pyrrolo [2,3-d] pyrimidin-7-yl) methyl Pivalate ( (R)-45 ) . Racemic (+/-)45 (151 mg) was dissolved in acetonitrile to a concentration of 30 mg/mL and a Daicel ChiralPak AD column (20 x 250 ) using 1000 μL of compound (+/-)45 solution per injection mm, 10 μm) by preparative HPLC using isocratic method: 30% isopropanol (+ 0.1% diethylamine)/70% hexanes (+ 0.1% diethyl) at a flow rate of 17 mL/min. amines). Baseline separations were achieved under these conditions with compound (S)-45 eluting at 15.5 min and compound (R)-45 eluting at 20.7 min.

Fractions containing each enantiomer were pooled separately and concentrated to give 51 mg of compound (S)-45 as a colorless film and 53 mg of compound (R)-45 as a colorless film.

Step 8. (R)-3-(4-(7H -pyrrolo[2,3-d]pyrimidin- 4-yl)-1H- pyrazol -1-yl)-3-(3,3,4 ,4-d ₄ -Cyclopentyl) propanenitrile (Compound 103 ) . (R)-45 (53 mg, 0.13 mmol, 1 eq) was dissolved in methanol (2 mL) in a 20 mL scintillation vial. Sodium hydroxide (0.25 mL of a 1M solution, 0.25 mmol, 2 equiv) was added and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was diluted with water (10 mL) and brine (20 mL). The aqueous mixture was extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated. The crude material was purified using an Analogix automated chromatography system eluting with 0-6% methanol in dichloromethane. Product fractions were pooled and evaporated to give compound 103 as a white foam with ˜90% purity with incompletely deprotected hydroxymethyl intermediate as the major impurity. Further chromatography did not further improve the purity. The 90% pure material was dissolved in THF (2 mL) and treated with several drops of 10% aqueous sodium hydroxide at 40° C. for 8 hours to effect complete conversion to compound 103. The resulting mixture was diluted with water (10 mL) and extracted with ethyl acetate (2×10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated to a white foam. The foam was dissolved in minimal acetonitrile, diluted with water and lyophilized to give compound 103 (14 mg, 35% yield) as a white solid. Chiral purity >99% ee (Chiralpak OD 4.6 x 250 mm, 10 um, 70% (hexane + 0.1% diethylamine) + 30% (isopropanol + 0.1% diethylamine), 1 mL/min, 254 nm residence time = 7.56 min).

Example 3. (R)-3-(4-(7H -pyrrolo[2,3-d]pyrimidin- 4-yl)-1H- pyrazol -1-yl)-3-( cyclopentyl - d9 ) propane Synthesis of Nitrile (Compound 127 ).

Scheme 5. Preparation of compound 127

Step 1. Diethyl 2,2,3,3,4,4,5,5-d ₈ -cyclopentane-1,1-dicarboxylate (47). To a solution of diethyl malonate (6.24 mL, 41.1 mmol) in ethanol (40 mL) was added a 21 wt % solution of sodium ethoxide in ethanol (30.7 mL, 82.2 mmol) followed by 1,1,2,2,3 ,3,4,4-octadeutero-1,4-dibromobutane (46, 9.67 g, 43.2 mmol, CDN isotope, 98 atomic % D) was added. The resulting solution was stirred under reflux for 2 hours, then cooled to room temperature and diluted with excess water. Most of the ethanol was then removed through distillation, and the obtained aqueous solution was extracted with ethyl acetate (3 x 75 mL). The organic layers were combined, washed with brine, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give 47 as a yellow oil (9.12 g, 100%), which was used in the next step without purification. did.

Step 2. Purdeuterocyclopentane-1-carboxylic acid (48). To a solution of 47 (9.12 g, 41.1 mmol) in ethanol (20 mL) was added 5M sodium hydroxide solution (20 mL). Additional water (15 mL) was then added and the reaction stirred at reflux for 3 h. Upon cooling to room temperature, the reaction was diluted with excess water and most of the ethanol was removed via distillation. The aqueous solution was made acidic (pH<2) with 1N HCl, then extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The resulting pale orange solid was transferred to a pressure flask and D ₂ O (120 mL) was added. The pressure flask was sealed and the reaction stirred at 160° C. for 15 h, then cooled to room temperature. The reaction was diluted with 1N HCl and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 48 (4.58 g, 90%) as a yellow oil, which was used without purification.

Step 3. N-Methoxy-N-methyl(cyclopentane-d ₉ )carboxamide (49). To a solution of 48 (4.58 g, 37.2 mmol) in acetonitrile (60 mL) at 0° C. N,O-dimethylhydroxylamine hydrochloride (4.35 g, 44.6 mmol), TBTU (12.5 g, 39.1 mmol) and N,N-diisopropylethylamine (19.4 mL, 112 mmol). The reaction was stirred at room temperature for 15 h, then diluted with 1N HCl and extracted with ethyl acetate (3×50 mL). The organic layers were combined, washed with saturated NaHCO ₃ , dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The obtained product was purified by column chromatography (SiO ₂ , 0-50% ethyl acetate/hexanes) to give compound 49 (3.41 g, 55%) as a clear oil. MS (ESI) 167.2 [(M + H) ⁺ ].

Step 4. Purdeuterocyclopentane-1-carboxaldehyde (50). To a solution of compound 49 (3.41 g, 20.5 mmol) in THF (80 mL) at 0° C. was added 1M LiAlH ₄ solution in THF (37.0 mL, 37.0 mmol) dropwise. The reaction was stirred at room temperature for 1 h, then quenched at 0° C. by sequential dropwise addition of _{D 2} O (1.41 mL), 15% NaOD/D ₂ O (1.41 mL) and D _{2 O (4.23 mL).} The quenched reaction was stirred at room temperature for 30 min, then filtered through Celite® and concentrated under reduced pressure. The resulting oil was diluted with 1N DCl/D ₂ O and extracted with diethyl ether (3×50 mL). The organic layers were combined, dried (MgSO ₄ ), filtered and concentrated under reduced pressure to give compound 50 (1.79 g, 82%) as a clear oil, which was used without purification.

Step 5. 3-(purdeuterocyclopentyl)acrylonitrile (51). To a solution of diethyl cyanoethylphosphonate (1.35 mL, 8.34 mmol) in THF (25 mL) at 0° C. was added 1M potassium tert-butoxide solution in THF (8.34 mL, 8.34 mmol) dropwise. The reaction was stirred at 0° C. for 1 h. Aldehyde (50) (1.79 g, 16.7 mmol) was then added dropwise as a solution in THF (5 mL). The reaction was stirred at room temperature for 15 h, then diluted with an excess of 1:1 water/brine and extracted with MTBE (3×50 mL). The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. The organic layers were combined, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure to give compound 51 (1.61 g, 74%) as a pale orange oil, which was used without purification.

Step 6. (+/-)-(4-(1-(2-cyano-1-(cyclopentyl-d ₉ )ethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)methyl pivalate ((+/-)52). In a solution of compound 37 (619 mg, 2.07 mmol, the preparation described in Lin, Q. et al. Org. Lett., 2009, 11, 1999-2002) in acetonitrile (15 mL), compound 51 (673) mg, 5.17 mmol) followed by DBU (650 μl, 4.35 mmol). The reaction was stirred at room temperature for 15 h, then concentrated under reduced pressure. The resulting crude mixture was diluted with water and extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, washed with 1N HCl, dried (Na ₂ SO ₄ ), filtered and concentrated under reduced pressure. Purification via normal phase column chromatography (SiO ₂ , 0-60% ethyl acetate/hexanes) gave ((+/-)52) (447 mg, 50%) as a white foam. 1H NMR (DMSO-d ₆ , 400 MHz) δ 8.84 (s, 1H), 8.79 (s, 1H), 8.39 (s, 1H), 7.75 (d, J = 3.7 Hz, 1H), 7.12 (d, J) = 3.7 Hz, 1H), 6.24 (s, 2H), 4.53 (dd, J = 9.6, 4.2 Hz, 1H), 3.32 - 3.13 (m, 2H), 1.08 (s, 9H).; MS (ESI) 430.3 [(M + H) ⁺ ].

Step 7. (R)-(4-(1-(2-cyano-1-(cyclopentyl-d ₉ )ethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3- d]pyrimidin-7-yl)methyl pivalate ((R)-52). Racemic compound ((+/-)52) (162 mg) was dissolved in acetonitrile to a concentration of 30 mg/mL and injected using the isocratic method on a Daicel ChiralPak AD column (20 x 250 mm, 10 μm). chiral separation by preparative HPLC with 1000 μl of compound (+/-)52 solution per sugar: 30% isopropanol (+ 0.1% diethylamine)/ 70% hexanes (+ 0.1% diethylamine) at a flow rate of 17 mL/min . Under these conditions, the baseline separation was achieved with compound (S)-52 eluting at 15.4 min and compound (R)-52 eluting at 20.5 min.

Fractions containing each enantiomer were collected separately and concentrated to give 61 mg of compound ((S)-52) as a colorless film and 63 mg of compound ((R)-52) as a colorless film.

Step 8. (R)-3-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl)-3-(cyclopentyl-d ₉ ) propanenitrile (compound 127). Compound (R)-52 (60 mg, 0.14 mmol, 1 equiv) was dissolved in methanol (2 mL) in a 20 mL scintillation vial. Sodium hydroxide (0.28 mL of a 1 M solution, 0.28 mmol, 2 equiv) was added and the reaction mixture was stirred at room temperature for 18 h. The reaction mixture was diluted with water (10 mL) and brine (20 mL). The aqueous mixture was extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated. The crude material was purified using an Analogix automated chromatography system eluting with 0-6% methanol in dichloromethane. Product fractions were pooled and evaporated to give compound 127 (34 mg) as a white foam in -90% purity with incompletely deprotected hydroxymethyl intermediate as the major impurity. Additional chromatography failed to further improve the purity. The 90% pure material was dissolved in THF (2 mL) and treated with several drops of 10% aqueous sodium hydroxide at 40° C. for 8 hours to complete conversion to compound 127. The reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (2×10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated to a white foam. The foam was dissolved in minimal acetonitrile, diluted with water and lyophilized to give compound 127 (19 mg, 42% yield) as a white solid. Chiral purity >99% ee (Chiralpak OD 4.6 x 250 mm, 10 μm, 70% (hexane + 0.1% diethylamine) + 30% (isopropanol + 0.1% diethylamine), 1 mL/min, 254 nm retention time = 7.55 min).

Example 4. Evaluation of Metabolic Stability in ^{CYP3A4 Supersomes TM}

Evaluation of the metabolic stability of compounds 103, 107 and 127 in human CYP3A4 Supersomes ^™

SUPERSMES ^TM Assay. Stock solutions of 10 mM test compounds 103, 107, 127 and ruxolitinib were prepared in DMSO. 10 mM stock solutions were diluted to 15.6 μM in acetonitrile (ACN). Human CYP3A4 supersomes ^™ (1000 pmol/mL, ^{purchased from BD Gentest™} Products and Services) was diluted to 62.5 pmol/mL in 0.1 M potassium phosphate buffer containing _{3 mM MgCl 2 , pH 7.4.} Diluted supersomes were added in triplicate to the wells of a 96-well polypropylene plate. A 10 μl aliquot of 15.6 μM test compound was added to the supersomes and the mixture was pre-warmed for 10 minutes. The reaction was initiated by addition of a 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 MgCl 2 . The reaction mixture was incubated at 37° C., 50 μl aliquots removed at 0, 5, 10, 20, and 30 min, and 96 containing 50 μl ice-cold ACN with internal standard to stop the reaction. -Added to well plate. The plate was stored at 4° C. for 20 minutes, after which 100 mL of water was added to the wells and centrifuged to pellet the precipitated proteins. The supernatant was transferred to another 96-well plate and analyzed for the amount of residual parent compound by LC-MS/MS using an Applied Bio-systems API 4000 mass spectrometer.

Data analysis: was calculated from the slope of the linear regression of the in vitro half-life for the test compound (t _1/2 value) for LN (residual% of the parent compound) versus incubation time relationship:

In vitro t _½ = 0.693/k, where k = -[slope of linear regression of % parent compound remaining (ln) versus incubation time].

The results of these experiments are shown in Table 3 and FIG. 1 . As described in Table 3, the half-life of ruxolitinib was calculated to be 14.5 minutes. On the other hand, compounds 103, 107 and 127 were more stable in supersomes, respectively, with calculated half-lives of 16.9, 17.9 and 32.0 min, respectively. This represents a 17% increase, 121% increase in the t _1/2 In the case of t 23% increase in _1/2, and in the case of compound 127 Compound 107 of t _1/2 In the case of compound 103.

Table 3: Metabolic stability of compounds 103, 107 and 127 versus ruxolitinib in human Human CYP3A4 Supersomes ^™

Example 5. Evaluation of Metabolic Stability in Human Liver Micro

Microsome Assay: Human liver microsomes (20 mg/mL) were obtained from Xenotech, LLC (Lenexa, KS). β-Nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), magnesium chloride (MgCl ₂ ), and dimethyl sulfoxide (DMSO) were purchased from Sigma Aldrich.

Determination of metabolic stability: 7.5 mM stock solutions of test compounds were prepared in DMSO. The 7.5 mM stock solution was diluted to 12.5-50 μM in acetonitrile (ACN). 20 mg/mL human liver microsomes were diluted to 0.625 mg/mL in 0.1 M potassium phosphate buffer containing _{3 mM MgCl 2 , pH 7.4.} Diluted microsomes were added in triplicate to wells of 96-well deep-well polypropylene plates. A 10 μl aliquot of 12.5-50 μM test compound was added to the microsomes and the mixture was pre-warmed for 10 minutes. The reaction was initiated by addition of a pre-warmed NADPH solution. The final reaction volume was 0.5 mL and contained 0.5 mg/mL human liver microsomes, 0.25-1.0 μM test compound, and 2 mM NADPH in _{0.1 M potassium phosphate buffer, pH 7.4 and 3 mM MgCl 2 .} The reaction mixture was incubated at 37° C., 50 μl aliquots removed at 0, 5, 10, 20, and 30 min, and shallow containing 50 μl ice-cold ACN with internal standard to stop the reaction. -Added to 96-well plates of wells. The plate was stored at 4° C. for 20 minutes, after which 100 μl of water was added to the wells of the plate and centrifuged to pellet the precipitated protein. The supernatant was transferred to another 96-well plate and analyzed for the amount of residual parent compound by LC-MS/MS using an Applied Bio-systems API 4000 mass spectrometer. The same procedure was performed for the non-deuterated counterpart of the compound of formula (I) or formula (A) and the positive control, 7-ethoxycoumarin (1 μM). The test was performed three times.

Data analysis: was calculated from the slope of linear regression of% parent compound remaining in vitro t _1/2 for the test compound (ln) versus incubation time relationship:

In vitro t _½ = 0.693/k,

where k = -[slope of linear regression of residual parent compound % (ln) versus incubation time].

Data analysis was performed using Microsoft Excel software.

It is understood that, without further explanation, one skilled in the art, using the preceding description and illustrative examples, will be able to make, use, and practice the claimed methods of the compounds of the present invention. It is to be understood that the above discussion and examples merely set forth a detailed description of certain preferred embodiments. It will be apparent to those skilled in the art that various modifications and equivalents can be made without departing from the spirit and scope of the invention.

Claims (31)

A compound of formula (I): or a pharmaceutically acceptable salt thereof:

wherein Y ¹ is hydrogen;
each Y ² is independently selected from hydrogen and deuterium, provided that each Y ² bonded to a common carbon is the same;
each Y ³ is independently selected from hydrogen and deuterium, provided that each Y ³ bonded to a common carbon is the same;
Y ⁴ is selected from hydrogen and deuterium;
each Y ⁵ is the same and is selected from hydrogen and deuterium;
Y ⁶ , Y ⁷ , and Y ⁸ are each independently selected from hydrogen and deuterium;
provided that each Y ² is deuterium, each Y ³ is deuterium, or each Y ² and Y ³ is deuterium;
Each position designated as D (deuterium) has at least 90% incorporation of deuterium. The compound according to claim 1, wherein each Y ² is hydrogen, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein each Y ³ is hydrogen, or a pharmaceutically acceptable salt thereof. 4. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein each Y ^{5 is hydrogen.} The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Y ⁷ , and Y ^{8 are each hydrogen.} The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein Y ^{6 is hydrogen.} The compound according to any one of claims 1 to 3, wherein Y ⁴ is hydrogen, or a pharmaceutically acceptable salt thereof. The compound according to claim 1, wherein Y ⁶ , Y ⁷ , and Y ⁸ are each hydrogen, and the compound is selected from any one of the compounds (Cmpd) listed in the table below, or a pharmaceutically acceptable salt thereof:

Here, any atom not designated as deuterium is present in its natural isotopic abundance. The compound according to claim 1, wherein Y ⁶ , Y ⁷ , and Y ⁸ are each deuterium, and the compound is selected from any one of the compounds (Cmpd) listed in the table below, or a pharmaceutically acceptable salt thereof:

Here, any atom not designated as deuterium is present in its natural isotopic abundance. The compound according to claim 1, wherein the compound is selected from the group consisting of: or a pharmaceutically acceptable salt thereof:

,

, and

. 11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and 8 to 10, wherein each position designated as D (deuterium) has at least 95% incorporation of deuterium. 11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and 8 to 10, wherein each position designated as D (deuterium) has at least 97% incorporation of deuterium. 11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and 10, wherein any atom not designated as deuterium is present in its natural isotopic abundance. 11. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and 8 to 10, wherein the pharmaceutically acceptable salt of the compound is a phosphate salt. delete delete delete Claims 1 to 3 and 8 to 10, comprising the compound of any one of claims or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, myelofibrosis (myelofibrosis), pancreatic cancer, prostate cancer A pharmaceutical composition for treating a disease selected from the group consisting of prostate cancer, breast cancer, leukemia, non-Hodgkin's lymphoma, multiple myeloma, and psoriasis. 19. The medicament of claim 18, further comprising a therapeutic agent selected from lenalidomide, panobinostat, capecitabine, exemestane, and combinations thereof. academic composition. 19. The method of claim 18, wherein the myelofibrosis is primary myelofibrosis, post-polycythemia vera myelofibrosis, post-essential thrombocythemia myelofibrosis, essential platelets A pharmaceutical composition which is essential thrombocythemia or a combination thereof. According to claim 1, wherein the compound represented by the following structure, or a pharmaceutically acceptable salt thereof:

. 22. The compound or pharmaceutically acceptable salt thereof according to claim 21, wherein each position designated as D (deuterium) has at least 95% incorporation of deuterium. 22. The compound or pharmaceutically acceptable salt thereof according to claim 21, wherein each position designated as D (deuterium) has at least 97% incorporation of deuterium. 24. The compound or pharmaceutically acceptable salt thereof according to any one of claims 21 to 23, wherein any atom not designated as deuterium is present in its natural isotopic abundance. 24. The compound or a pharmaceutically acceptable salt thereof according to any one of claims 21 to 23, wherein the pharmaceutically acceptable salt of the compound is a phosphate salt. delete delete delete 24. Myelofibrosis, pancreatic cancer, prostate cancer, breast cancer, leukemia, non-Hodgkin's lymphoma, comprising a compound of any one of claims 21 to 23, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier; A pharmaceutical composition for treating a disease selected from the group consisting of multiple myeloma, and psoriasis. 30. The pharmaceutical composition of claim 29, further comprising a therapeutic agent selected from lenalidomide, panobinostat, capecitabine, exemestane, and combinations thereof. 30. The pharmaceutical composition of claim 29, wherein the myelofibrosis is primary myelofibrosis, myelofibrosis after polycythemia vera, myelofibrosis after essential thrombocytopenia, essential thrombocytopenia, or a combination thereof.

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