Canine Janus kinase inhibitor and application thereof

Abstract

Provided herein are compounds having inhibitory activity on the Janus kinase 1 (JAK-1) enzyme, which are useful as therapeutic and / or prophylactic agents. The compounds may be used in the treatment of inflammatory disorders in non-human mammals, for example, in the treatment of canine atopic dermatitis. Also provided herein are methods for preparing such compounds.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 470,589, filed June 2, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0003] This disclosure generally relates to canine Janus kinase (JAK) inhibitors and methods of using such inhibitors to treat inflammatory disorders. Background Technology

[0004] Protein kinases are a family of enzymes that catalyze the phosphorylation of specific amino acid residues present in certain proteins. These protein kinases are broadly classified into tyrosine kinases and serine / threonine kinases. Inappropriate kinase activity—which may arise from, for example, mutations, overexpression, or inappropriate regulation, as well as overexpression or underexpression of growth factors or cytokines—has been identified as being associated with a wide range of diseases. These diseases include, but are not limited to, cancer, cardiovascular disease, allergies, asthma and other respiratory diseases, autoimmune diseases, inflammatory diseases, bone diseases, metabolic disorders, and neurodegenerative disorders. Inappropriate kinase activity triggers a variety of cellular responses related to cell growth, differentiation, survival, apoptosis, and mitosis, as well as cell cycle control and cell movement, each of which is associated with the aforementioned diseases.

[0005] Therefore, protein kinases have become important target enzymes for therapeutic interventions. In particular, Janus kinases (JAKs) are a family of cellular protein tyrosine kinases that play a central role in cytokine signaling (Kisseleva et al., Gene 2002, 285, 1; Yamaoka et al., Genome Biology 2004, 5, 253). JAKs function as dimers in the signaling processes of many cytokine receptors. JAKs comprise four family members: JAK-1, JAK-2, JAK-3, and tyrosine kinase 2 (TYK-2). Many cytokines are known to activate the JAK family. Upon binding to their receptors, cytokines activate JAKs, which then phosphorylate the cytokine receptors, creating docking sites for signaling molecules. These signaling molecules include members of the signal transducer and activator of transcription (STAT) family, which ultimately lead to gene expression.

[0006] JAK plays a key role in both innate and adaptive immunity, making it an attractive target for the treatment of inflammatory diseases. For autoimmune diseases, targeting the JAK signaling pathway is supported by the involvement of multiple proinflammatory cytokines that signal through the JAK pathway in the pathogenesis of these immune-related disorders. Activation of JAK signaling initiates the expression of survival factors, cytokines, chemokines, and other molecules that promote leukocyte trafficking and cell proliferation, leading to inflammatory and autoimmune disorders. (O'Shea et al., N Engl J Med. 2013, 368(2), 161-70).

[0007] In view of the potential of JAK inhibitors to target pathways associated with a variety of diseases and disorders, including inflammatory disorders, there is a need in the art to provide additional JAK inhibitors and methods of utilizing such inhibitors in the treatment of diseases and disorders that respond to such inhibitors. In particular, there is a clear unmet need for new pharmaceutical agents to control atopic dermatitis in animals that can address certain limitations of existing therapeutic agents. SUMMARY

[0008] The present disclosure relates to compounds that are inhibitors of Janus kinases (JAKs), have efficacy against canine Janus kinase-1 (cJAK-1), and are selective relative to canine Janus kinase-2 (cJAK-2), canine Janus kinase-3 (cJAK-3), and canine tyrosine kinase 2 (cTYK-2). Accordingly, the disclosed compounds can be used as therapeutic agents for indications requiring immunosuppression and / or immunomodulation, including but not limited to canine atopic dermatitis.

[0009] Atopic dermatitis (AD; also known as atopic eczema) is an inflammatory, pruritic, chronic or chronically relapsing skin disease with a genetic predisposition. It is most commonly associated with IgE antibodies to environmental allergens. Common clinical features include erythema, edema, dryness, erosion / excoriation, weeping, and crusting. The disease typically affects dogs aged 6 months to 3 years, characterized by pruritus and secondary skin lesions, characteristically distributed around the face (mouth, eyes), concave surface of the pinnae, ventral abdomen, flexor surfaces of the elbows, wrists, and hock joints, interdigital skin, and / or perineal area. Animals with atopic dermatitis are prone to secondary skin infections, ear infections, and yeast infections. Atopic dermatitis cannot be cured. Therefore, the goal is to control the disease to improve the quality of life for the dog and its owner.

[0010] The treatment market for atopic dermatitis in animals has historically been dominated by corticosteroids and antihistamines, but each has various shortcomings. For example, corticosteroids can produce adverse side effects in animals, particularly in companion animals such as dogs, while antihistamines are less efficacious. In particular, short- and long-term side effects of corticosteroids include polydipsia, polyphagia, polyuria, pancreatitis, gastrointestinal ulceration, lipemia, diabetes mellitus, muscle atrophy, and iatrogenic Cushing's syndrome. In addition, complex dosing regimens can be challenging for dogs and their owners. One canine formulation of cyclosporine (ATOPICATM) has been marketed for atopic dermatitis, but is expensive, slow to take effect, and has issues with gastrointestinal tolerability. In 2013, the FDA approved APOQUEL ® (Olacitinib; N-methyl[trans-4-(methyl-7H-pyrrolo[2,3-d]pyrimidin-4-ylamino)cyclohexyl]methanesulfonamide (2Z)-2-butenedioate; Zoetis) for the treatment of pruritus associated with allergic dermatitis and control of atopic dermatitis in dogs at least 12 months of age. APOQUEL ® is a relatively selective JAK-1 and JAK-3 inhibitor, thereby blocking the action of inflammatory cytokines (IL-2, IL-4, IL-6, IL-13) released by activated lymphocytes, as well as IL-31, a cytokine directly involved in the sensation of itch. Despite commercial success, APOQUEL ® requires twice daily oral dosing during an initial induction phase, which lasts up to two weeks. Given the challenges associated with oral dosing in companion animals, there is a need to provide an alternative therapeutic agent with properties that allow once daily dosing.

[0011] The compounds of the present disclosure are JAK inhibitors that are potent, efficacious, and selective for human JAK-1. Given the very high homology between canine and human JAKs, this potency and selectivity obtained for the human isoform is a suitable surrogate indicator for the canine isoform, and it is expected that the in vitro pharmacology profile in the canine will mirror that in humans. The activity profile and selectivity for JAK-1 can allow once daily dosing, while providing desirable efficacy and tolerability. Thus, these compounds represent a valuable alternative to existing therapeutic agents for the treatment of chronic pruritus and inflammation associated with atopic dermatitis. Accordingly, the present disclosure provides compounds of Formula I, their use as cJAK-1 inhibitors for the treatment of canine atopic dermatitis, pharmaceutical compositions containing these compounds, and methods for preparing these compounds.

[0012] In one aspect, there is provided a compound having a structure according to Formula (I):

[0013] (I),

[0014] or a pharmaceutically acceptable salt or solvate thereof, wherein:

[0015] R1is H or optionally substituted C1-C3alkyl;

[0016] n = 0 or 1 ;

[0017] R2is H or CH3;

[0018] R3is C1-C4alkyl, phenyl,

[0019] , or ,

[0020] wherein:

[0021] m is 0 or 1 ;

[0022] X is H, F, CI, OCH3, or CH3;

[0023] R4is H or CH3;

[0024] R5is H, C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, or 2-pyrimidyl; or

[0025] R4and R5together with the included N atom form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally includes one additional heteroatom selected from N and O; or

[0026] R2and R3together with the included N atom form a 6-membered ring, which is optionally substituted with C1-C4alkyl, C7to C 12 aralkyl or -C(O)-NR6R7,

[0027] wherein:

[0028] R6is H or C1-C4alkyl;

[0029] R7is H, C1-C4alkyl, phenyl; or

[0030] R6and R7together with the included N atom form a 6-membered ring, which can be optionally substituted with C1-C4alkyl, and wherein the 6-membered ring optionally includes one additional N atom.

[0031] In some embodiments, R2is H and R3is:

[0032] .

[0033] In some embodiments, n is 1 and R1is H.

[0034] In some embodiments, n is 1 and R1is CH3.

[0035] In some embodiments, R3is:

[0036] .

[0037] In some embodiments, m is 0.

[0038] In some embodiments, R4and R5are each H.

[0039] In some embodiments, the compound of Formula I is selected from:

[0040] and .

[0041] In some embodiments, m is 1.

[0042] In some embodiments, R4and R5are each CH3.

[0043] In some embodiments, the compound of Formula I is:

[0044] .

[0045] In some embodiments:

[0046] R1is H;

[0047] R2is H;

[0048] R4is H; and

[0049] R5is selected from C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidyl.

[0050] In some embodiments:

[0051] R1is CH3;

[0052] R2is H;

[0053] R4is H; and

[0054] R5is selected from C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidyl.

[0055] In some embodiments, the compound of Formula I is selected from:

[0056] .

[0057] In some embodiments:

[0058] R1is H or CH3;

[0059] R2is H; and

[0060] R4and R5are each independently selected from the group consisting of C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidyl.

[0061] In some embodiments, the compound of Formula I is selected from:

[0062] .

[0063] In some embodiments:

[0064] R1is H or CH3;

[0065] R2is H; and

[0066] R4and R5together with the included N atom form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally includes one additional heteroatom selected from N and O;

[0067] In some embodiments, the compound of Formula I is selected from:

[0068] .

[0069] In some embodiments, the compound of Formula I is:

[0070] , , or .

[0071] In some embodiments, the compound of Formula I is:

[0072] , , or .

[0073] In some embodiments, R2and R3together with the included N atom form a 6-membered ring, which is optionally substituted with C1-C4alkyl, C7to C 12 aralkyl, or -C(O)-NR6R7.

[0074] In some embodiments, the compound of Formula I has a structure according to Formula II:

[0075] (II),

[0076] wherein Z is C or N.

[0077] In some embodiments, the compound of Formula II has a structure selected from:

[0078] .

[0079] In some embodiments, the compound of Formula I has the following structure:

[0080] .

[0081] In another aspect, a pharmaceutical composition is provided comprising a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof.

[0082] In still another aspect, a method is provided for treating an allergic reaction, allergic dermatitis, atopic dermatitis, eczema, or pruritus in a mammal comprising administering to a mammal in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, or a composition of any of the foregoing.

[0083] In some embodiments, the mammal is a companion animal. In some embodiments, the companion animal is a dog.

[0084] In some embodiments, the compound of Formula I is administered orally, parenterally, or topically. In some embodiments, the compound of Formula I is administered orally once daily.

[0085] These and other features, aspects, and advantages of the present disclosure will become evident to those of ordinary skill in the art from a reading of the following detailed description. The present disclosure includes any combination of two, three, four, or more of the features or elements set forth in the present disclosure, whether or not those features or elements are explicitly combined or listed in a particular example embodiment described herein. The present disclosure is intended to be construed as a whole, such that any separable feature or element of the present disclosure, in any of its aspects and example embodiments, should be considered combinable unless the context of the present disclosure explicitly indicates otherwise. Therefore, it is to be understood that the present disclosure is provided for the purpose of summarizing some example embodiments only and to provide a basic understanding of some aspects of the present disclosure. It is not intended to be a comprehensive or exhaustive presentation of the present disclosure nor is it intended to be a listing of elements or features that must be included in the practice of the present disclosure. Therefore, it is to be understood that the example embodiments described above are merely examples and should not be construed as limiting the scope or spirit of the present disclosure in any way. Other example embodiments, aspects, and advantages will become apparent from the following detailed description. DETAILED DESCRIPTION

[0086] The present disclosure will now be described more fully hereinafter with reference to example embodiments thereof. Before describing the several embodiments of the technology, it is to be understood that the technology is not limited to the details of construction or process steps set forth in the following description. The technology is capable of other embodiments and of being practiced or being carried out in various ways.

[0087] The following description sets forth numerous specific configurations, methods, parameters, and the like, in an effort to provide a thorough understanding of various embodiments of the disclosure. It should be recognized, however, that such description is not intended as limiting the scope of the disclosure, but rather as a description of exemplary embodiments.

[0088] The present disclosure relates generally to compounds of Formula I:

[0089] (I),

[0090] including stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof, and pharmaceutical compositions comprising the compounds or stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof. The present disclosure is particularly directed to such compounds having canine Janus kinase-1 (cJAK-1) inhibitory activity, stereoisomers, tautomers, solvates, and pharmaceutically acceptable salts thereof. Such compounds and pharmaceutical compositions of the present disclosure can be used to treat canine atopic dermatitis with its associated chronic pruritus and inflammation. The compounds, compositions, and methods of treatment are further described below.

[0091] Definitions

[0092] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. In connection with the terms used in the present disclosure, the following definitions are provided. Unless the text of the definition of a term requires a different meaning in a specific context, the term used in this application will be defined as follows.

[0093] The articles “a” and “an” as used in the present disclosure can refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” can mean one element or more than one element.

[0094] The term “and / or” as used in the present disclosure can mean “and” or “or”, unless otherwise indicated.

[0095] The term “about” as used in the present specification is used to describe and account for small fluctuations. For example, the term “about” can mean less than or equal to ± 10%, less than or equal to ± 5%, less than or equal to ± 2%, less than or equal to ± 1%, less than or equal to ± 0.5%, less than or equal to ± 0.2%, less than or equal to ± 0.1%, or less than or equal to ± 0.05%. All numerical amounts herein are modified by the term “about”, whether or not explicitly indicated. Values modified by the term “about” certainly include the specific values. For example, “about 5.0” must include 5.0.

[0096] Unless the context otherwise requires, the terms “comprising,” “containing,” and “including” are used in their open, non-limiting sense in this specification and claims as used herein.

[0097] "Alkyl" refers to a straight-chain or branched hydrocarbon chain consisting only of carbon and hydrogen atoms, without unsaturated bonds, and preferably having 1 to 15 carbon atoms (i.e., C1-C1). 15 Alkyl groups. In some embodiments, the alkyl group comprises 1 to 13 carbon atoms (i.e., C1-C1). 13 Alkyl group. In some embodiments, the alkyl group comprises 1 to 8 carbon atoms (i.e., C1-C8 alkyl). In other embodiments, the alkyl group comprises 1 to 5 carbon atoms (i.e., C1-C5 alkyl). In other embodiments, the alkyl group comprises 1 to 4 carbon atoms (i.e., C1-C4 alkyl). In other embodiments, the alkyl group comprises 1 to 3 carbon atoms (i.e., C1-C3 alkyl). In other embodiments, the alkyl group comprises 1 to 2 carbon atoms (i.e., C1-C2 alkyl). In other embodiments, the alkyl group comprises 1 carbon atom (i.e., C1 alkyl). In other embodiments, the alkyl group comprises 5 to 15 carbon atoms (i.e., C5-C6 alkyl). 15 Alkyl group. In other embodiments, the alkyl group comprises 5 to 8 carbon atoms (i.e., C5-C8 alkyl). In other embodiments, the alkyl group comprises 2 to 5 carbon atoms (i.e., C2-C5 alkyl). In other embodiments, the alkyl group comprises 3 to 5 carbon atoms (i.e., C3-C5 alkyl). In some embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (isopropyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). The alkyl group is connected to the rest of the molecule by a single bond.

[0098] When used in conjunction with a chemical moiety (such as alkyl, alkenyl, or ynyl), the term "C" is used. x-y "This refers to groups that contain x to y carbons in the chain. For example, the term "C" 1-6 "Alkyl" refers to a substituted or unsubstituted saturated hydrocarbon group containing 1 to 6 carbon atoms, including straight-chain alkyl and branched-chain alkyl groups. Term -C x-y Alkylene refers to an alkylene chain having x to y carbons, whether substituted or unsubstituted. For example, -C 1-6 The alkylene group can be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any of which may be optionally substituted.

[0099] "Alkoxy" refers to a group bonded by an oxygen atom of the formula -O-alkyl, wherein the alkyl is an alkyl chain as defined above.

[0100] "Alkenyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing at least one carbon to carbon double bond, and preferably having from 2 to 12 carbon atoms (i.e., C2-C 12 Alkenyl). In certain embodiments, an alkenyl comprises 2 to 8 carbon atoms (i.e., C2-C8 alkenyl). In certain embodiments, an alkenyl comprises 2 to 6 carbon atoms (i.e., C2-C6 alkenyl). In other embodiments, an alkenyl comprises 2 to 4 carbon atoms (i.e., C2-C4 alkenyl). The alkenyl is attached to the remainder of the molecule by a single bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e., allyl), but-1-enyl, pent-1-enyl, pent-1,4-dienyl, and the like.

[0101] "Alkynyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing at least one carbon to carbon triple bond, and preferably having from 2 to 12 carbon atoms (i.e., C2-C 12 Alkynyl). In certain embodiments, an alkynyl comprises 2 to 8 carbon atoms (i.e., C2-C8 alkynyl). In other embodiments, an alkynyl comprises 2 to 6 carbon atoms (i.e., C2-C6 alkynyl). In other embodiments, an alkynyl comprises 2 to 4 carbon atoms (i.e., C2-C4 alkynyl). The alkynyl is attached to the remainder of the molecule by a single bond, for example, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0102] The terms "C x-y Alkenyl" and "C x-y Alkynyl" refer to substituted or unsubstituted unsaturated aliphatic groups, which are analogous to the alkyl groups described above in terms of length and possible substitution, but which contain at least one double or triple bond, respectively. The term -C x-y Alkenylene- refers to a substituted or unsubstituted alkenylene chain having x to y carbons in the alkenylene chain. For example, -C 2-6 Alkenylene- can be selected from the group consisting of ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any of which is optionally substituted. The alkenylene chain can have one double bond or more than one double bond in the alkenylene chain. The term -C x-y Alkynylene- refers to a substituted or unsubstituted alkynylene chain having x to y carbons in the alkenylene chain. For example, -C 2-6 Alkynylene- can be selected from the group consisting of ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any of which is optionally substituted. The alkynylene chain can have one triple bond or more than one triple bond in the alkynylene chain.

[0103] "alkylene" or "alkylene chain" refers to a straight-chain or branched divalent hydrocarbon chain in which the remainder of a molecule is attached to a group. It consists only of carbon and hydrogen, contains no unsaturated bonds, and preferably has 1 to 12 carbon atoms, such as methylene, ethylene, propylene, n-butylene, etc. The alkylene chain is attached to the remainder of the molecule and to the group via single bonds. The connection points between the alkylene chain and the remainder of the molecule and to the group can be through any two carbons within the chain. In some embodiments, the alkylene chain contains 1 to 10 carbon atoms (i.e., C64 ... 1- In some embodiments, the alkylene oxide comprises 1 to 8 carbon atoms (i.e., C1-C8 alkylene oxide). In other embodiments, the alkylene oxide comprises 1 to 5 carbon atoms (i.e., C1-C5 alkylene oxide). In other embodiments, the alkylene oxide comprises 1 to 4 carbon atoms (i.e., C1-C4 alkylene oxide). In other embodiments, the alkylene oxide comprises 1 to 3 carbon atoms (i.e., C1-C3 alkylene oxide). In other embodiments, the alkylene oxide comprises 1 to 2 carbon atoms (i.e., C1-C2 alkylene oxide). In other embodiments, the alkylene oxide comprises 1 carbon atom (i.e., C1 alkylene oxide). In other embodiments, the alkylene oxide comprises 5 to 8 carbon atoms (i.e., C5-C8 alkylene oxide). In other embodiments, the alkylene oxide comprises 2 to 5 carbon atoms (i.e., C2-C5 alkylene oxide). In other embodiments, the alkylene oxide comprises 3 to 5 carbon atoms (i.e., C3-C5 alkylene oxide).

[0104] "Alkenyl" or "alkenyl chain" refers to a straight-chain or branched divalent hydrocarbon chain that connects the rest of a molecule to a group, consisting only of carbon and hydrogen, containing at least one carbon-carbon double bond, and preferably having 2 to 12 carbon atoms. The alkenyl chain is connected to the rest of the molecule and to the group via single bonds. The connection points between the alkenyl chain and the rest of the molecule and with the group can be through any two carbons in the chain. In some embodiments, the alkenyl group contains 2 to 10 carbon atoms (i.e., C2-C). 10 (Alkenyl group). In some embodiments, the alkenyl group comprises 2 to 8 carbon atoms (i.e., C2-C8 alkenyl). In other embodiments, the alkenyl group comprises 2 to 5 carbon atoms (i.e., C2-C5 alkenyl). In other embodiments, the alkenyl group comprises 2 to 4 carbon atoms (i.e., C2-C4 alkenyl). In other embodiments, the alkenyl group comprises 2 to 3 carbon atoms (i.e., C2-C3 alkenyl). In other embodiments, the alkenyl group comprises 2 carbon atoms (i.e., C2 alkenyl). In other embodiments, the alkenyl group comprises 5 to 8 carbon atoms (i.e., C5-C8 alkenyl). In other embodiments, the alkenyl group comprises 3 to 5 carbon atoms (i.e., C3-C5 alkenyl).

[0105] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a group, which consists solely of carbon and hydrogen, contains at least one carbon-carbon triple bond, and optionally, one or more double bonds, and preferably has 2 to 12 carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to the group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the group can be through any two carbons within the chain. In certain embodiments, an alkynylene comprises 2 to 10 carbon atoms (i.e., C2-C10 alkynylene). In certain embodiments, an alkynylene comprises 2 to 8 carbon atoms (i.e., C2-C8 alkynylene). In other embodiments, an alkynylene comprises 2 to 5 carbon atoms (i.e., C2-C5 alkynylene). In other embodiments, an alkynylene comprises 2 to 4 carbon atoms (i.e., C2-C4 alkynylene). In other embodiments, an alkynylene comprises 2 to 3 carbon atoms (i.e., C2-C3 alkynylene). In other embodiments, an alkynylene comprises 2 carbon atoms (i.e., C2 alkynylene). In other embodiments, an alkynylene comprises 5 to 8 carbon atoms (i.e., C5-C8 alkynylene). In other embodiments, an alkynylene comprises 3 to 5 carbon atoms (i.e., C3-C5 alkynylene). 10 "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule to a group, which consists solely of carbon and hydrogen, contains at least one carbon-carbon triple bond, and optionally, one or more double bonds, and preferably has 2 to 12 carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to the group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the group can be through any two carbons within the chain. In certain embodiments, an alkynylene comprises 2 to 10 carbon atoms (i.e., C2-C

[0106] "Aryl" refers to a radical derived by removal of a hydrogen atom from a ring carbon atom of an aromatic monocyclic or aromatic polycyclic hydrocarbon ring system. The aromatic monocyclic or aromatic polycyclic hydrocarbon ring system contains only hydrogen and carbon atoms and from 5 to 18 carbon atoms, wherein at least one ring in the ring system is aromatic, that is, it contains a ring of cyclic, delocalized (4n+2) π-electrons, according to Hückel theory. Ring systems from which the aryl radical is derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin, and naphthalene.

[0107] "Arylalkyl" refers to a radical of the formula -R c -aryl, wherein R c is an alkylene chain as defined above, for example methylene, ethylene, and the like.

[0108] "Arylalkenyl" refers to a radical of the formula -R d -aryl, wherein R d is an alkenylene chain as defined above. "Arylalkynyl" refers to a radical of the formula -R e -aryl, wherein R e is an alkynylene chain as defined above.

[0109] "Carbocyclyl" refers to a saturated, unsaturated or aromatic ring wherein each ring atom is carbon. Carbocyclyl can include 3- to 10-membered monocyclic, 6- to 12-membered bicyclic and 6- to 12-membered bridged rings. Each ring of the bicyclic carbocyclyl can be selected from saturated, unsaturated and aromatic rings. In some embodiments, the carbocyclyl is aryl. In some embodiments, the carbocyclyl is cycloalkyl. In some embodiments, the carbocyclyl is cycloalkenyl. In one exemplary embodiment, an aromatic ring (e.g., phenyl) can be fused to a saturated or unsaturated ring (e.g., cyclohexane, cyclopentane or cyclohexene). Any combination of saturated, unsaturated and aromatic bicyclic rings is included in the definition of carbocyclyl as long as the valency permits. Exemplary carbocyclyl groups include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl and naphthyl.

[0110] "Cycloalkyl" refers to a stable, fully saturated monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, including fused or bridged ring systems, preferably having from three to twelve carbon atoms. In certain embodiments, the cycloalkyl group contains three to ten carbon atoms. In other embodiments, the cycloalkyl group contains five to seven carbon atoms. The cycloalkyl group can be attached to the rest of the molecule through a single bond. Examples of monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptan-2-yl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptan-2-yl, and the like.

[0111] "Cycloalkenyl" refers to a stable, unsaturated, non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, including fused or bridged ring systems, preferably having from three to twelve carbon atoms and containing at least one double bond. In certain embodiments, the cycloalkenyl group contains three to ten carbon atoms. In other embodiments, the cycloalkenyl group contains five to seven carbon atoms. The cycloalkenyl group can be attached to the rest of the molecule through a single bond. Examples of monocyclic cycloalkenyl groups include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0112] "Cycloalkylalkyl" refers to a radical of the formula -R c - cycloalkyl, wherein R c is an alkylene chain as described above.

[0113] "Cycloalkylalkoxy" refers to a radical of the formula -O-R c - cycloalkyl, bonded through an oxygen atom, wherein R c is an alkylene chain as described above.

[0114] "Halo" or "halogen" means a halogen substituent, i.e., bromo, chloro, fluoro, and iodo.

[0115] As used herein, the term "haloalkyl" or "haloalkane" refers to an alkyl group as defined above, which is substituted with one or more halogen groups, such as trifluoromethyl, dichloromethyl, bromomethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc. In some embodiments, the alkyl portion of the fluoroalkyl group may optionally be further substituted. Examples of halogen-substituted alkanes (“haloalkanes”) include halomethanes (e.g., chloromethane, bromomethane, fluoromethane, iodomethane), dihalomethanes and trihalomethanes (e.g., chloroform, tribromomethane, trifluoromethane, triiodomethane), 1-haloethane, 2-haloethane, 1,2-dihaloethane, 1-halopropane, 2-halopropane, 3-halopropane, 1,2-dihalopropane, 1,3-dihalopropane, 2,3-dihalopropane, 1,2,3-trihalopropane, and any other suitable combination of alkanes (or substituted alkanes) with halogens (e.g., Cl, Br, F, I, etc.). When the alkyl group is substituted by more than one halogen group, each halogen can be chosen independently, for example, 1-chloro-2-fluoroethane.

[0116] "Fluoroalkyl" refers to an alkyl group as defined above that is substituted with one or more fluorine groups, such as trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc.

[0117] "Heterocycle" refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring in a bicyclic heterocycle can be selected from saturated, unsaturated, and aromatic rings. In some embodiments, the heterocycle is a heteroaryl group. In some embodiments, the heterocycle is a heterocyclic alkyl group. "Hypoheterocyclic group" refers to a divalent heterocycle that links the remainder of the molecule to a group.

[0118] "Heterocyclic alkyl" refers to a stable 3- to 12-membered non-aromatic cyclic group containing 2 to 12 carbon atoms and at least one heteroatom, wherein each heteroatom may be selected from N, O, Si, P, B, and S atoms. Heterocyclic alkyl groups may be selected from monocyclic or bicyclic groups, as well as fused or bridged ring systems. The heteroatom in a heterocyclic alkyl group may optionally be oxidized. If one or more nitrogen atoms are present, it may optionally be quaternized. Heterocyclic alkyl groups may be partially or fully saturated. Wherever the valence allows, the heterocyclic alkyl group is connected to the rest of the molecule through any atom of the heterocyclic alkyl group (such as any carbon or nitrogen atom of the heterocyclic alkyl group). Examples of heterocyclic alkyl groups include, but are not limited to: dioxolane, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolinyl, isothiazolinyl, isoxazolinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, oxazolinyl, piperidinyl, piperazinyl, 4-piperidinoneyl, pyrrolyl, pyrazolyl, quininecycloyl, thiazolinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0119] "Heterocyclic alkyl alkyl" refers to the formula -R c - Heterocyclic alkyl groups, wherein R c The heterocyclic alkyl group is an alkylene chain as defined above. If the heterocyclic alkyl group is a nitrogen-containing heterocyclic alkyl group, the heterocyclic alkyl group may optionally be attached to the alkylene chain at the nitrogen atom.

[0120] "Heteroaryl" or "aromatic heterocycle" refers to a group derived from a 3- to 12-membered aromatic ring group comprising 1 to 11 carbon atoms and at least one heteroatom, wherein each heteroatom may be selected from N, O, and S. As used herein, the heteroaryl ring may be selected from monocyclic or bicyclic rings and fused or bridged ring systems, wherein at least one ring in the ring system is aromatic, i.e., according to Hückel theory, it comprises a cyclic, delocalized (4n+2)π electron system. The heteroatom(s) in the heteroaryl group may optionally be oxidized. One or more nitrogen atoms (if present) may optionally be quaternized. Wherever the valence allows, the heteroaryl group may be linked to the remainder of the molecule by any atom of the heteroaryl group (such as a carbon or nitrogen atom of the heteroaryl group). Examples of heteroaryl groups include, but are not limited to, aziridinetrienyl, acridine, benzimidazolyl, benzoindolyl, 1,3-benzo[m]dioxacyclopentenyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzo[thiadiazolyl, benzo[] ... b[1,4]dioxacycloheptenyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxacyclohexyl, benzonaphthofuranyl, benzoxazolyl, benzom-dioxacyclopentenyl, benzodioxacyclohexenyl, benzopyranyl, benzopyranoneyl, benzofuranyl, benzofuranoneyl, benzothienyl (benzothienyl) hiophenyl), benzothiophene[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridyl, carbazole, cenolinyl, cyclopentano[d]pyrimidinyl, 6,7-dihydro-5H-cyclopentano[4,5]thiophene[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cenolinyl, 6,7-dihydro-5H-benzo[ [6,7]cycloheptano[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanoneyl, furano[3,2-c]pyridyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridazinyl, 5,6,7,8,9,10-hexahydrocyclooctano[d]pyridyl, isothiazolyl, imidazolyl, indazole, indole, indazole , isoindolyl, dihydroindolyl, isodihydroindolyl, isoquinolinyl, indeneyl, isoxazolyl, 5,8-methyl-5,6,7,8-tetrahydroquinazolinyl, naphridinyl, 1,6-naphridinoneyl, oxadiazolyl, 2-oxoazacycloheptatrienyl (2oxoazepinyl), oxazolyl, oxacyclopropane, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1 H -pyrrole, phenazinyl, phenothiazinyl, phenotoxazinyl, phthalazinyl, pteridinyl, purine, pyrrole, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrole, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thiopheno[2] The following are listed: [3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cycloheptano[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyridano[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl, and thiophenyl (i.e., thienyl). "X-membered heteroaryl" refers to the number of inner ring atoms in the ring, i.e., X. For example, a 5-membered heteroaryl ring or a 5-membered aromatic heterocycle has 5 inner ring atoms, such as triazoles, oxazoles, and thienyl.

[0121] "Heteroarylalkyl" refers to the formula -R c - A heteroaryl group, wherein R c The alkylene chain is as defined above. If the heteroaryl group is a nitrogen-containing heteroaryl group, it may optionally be attached to the alkylene chain at the nitrogen atom.

[0122] As used in this article, the term "amino" refers to -NH2.

[0123] The terms "hydroxyl" and "hydroxyl" refer to -OH.

[0124] As used in this article, the term "oxo" refers to the "=O" group. It may also be abbreviated as C(O) or C=O in this article.

[0125] The term "substituted" refers to a portion of a structure in which a substituent replaces one or more hydrogen atoms on a carbon atom or a substituted heteroatom (e.g., NH). It should be understood that "substitution" or "replaced by" includes the implicit condition that such substitution meets the permissible valence of the substituted atom and the substituent, and that the substitution produces a stable compound, i.e., a compound that does not spontaneously transform, for example, through rearrangement, cyclization, elimination, etc. In some embodiments, substitution refers to a portion having a substituent that replaces two hydrogen atoms on the same carbon atom, for example, replacing two hydrogen atoms on a single carbon atom with an oxo, imino, or thio group. As used herein, the term "substituted" is intended to include all permissible substituents in an organic compound. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents in organic compounds. Permissible substituents may be one or more, and may be the same or different for a suitable organic compound. For the purposes of this disclosure, heteroatoms (e.g., nitrogen) may have hydrogen substituents and / or any permissible substituents of the organic compounds described herein, which satisfy the valence of the heteroatoms.

[0126] In some embodiments, the substituents may include any substituents described herein, such as: halogen, hydroxyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a -R b -OC(O)-R a -R b -OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(Ra )2、-R b -C(O)R a -R b -C(O)OR a -R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a -R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, areneyl, arynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl and heteroarylalkyl, any of which may optionally be substituted by: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=NNH2), -R b -OR a -R b -OC(O)-R a -R b -OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a -R b -C(O)OR a -R b -C(O)N(R a )2、-Rb -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a -R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); where each R a Independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl, wherein each R a (As long as the valence allows) it may be optionally substituted with the following: alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thio (=S), cyano (-CN), nitro (-NO2), imino (=NH), oxime (=N-OH), hydrazine (=N-NH2), -R b -OR a -R b -OC(O)-R a -R b -OC(O)-OR a -R b -OC(O)-N(R a )2、-R b -N(R a )2、-R b -C(O)R a -R b -C(O)OR a -R b -C(O)N(R a )2、-R b -OR c -C(O)N(R a )2、-R b -N(R a )C(O)OR a-R b -N(R a )C(O)R a -R b -N(R a S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2); and where each R b Independently selected from straight-chain or branched alkylene, alkenyl, or ynylene chains, and each R c It is a straight-chain or branched alkylene, alkenyl, or ynylene chain.

[0127] As used herein, the term “unsubstituted” means that the specified group is not substituented except for the stated portion (e.g., the valence is satisfied by hydrogen).

[0128] "Isomers" are different compounds having the same molecular formula. "Stereoisomers" are isomers that differ only in the spatial arrangement of their atoms. "Enantiomers" are a pair of stereoisomers that are non-overlapping mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture. The term "(±)" is used where appropriate to denote racemic mixtures. "Diadiamers" or "diastereomers" are stereoisomers having at least two asymmetric atoms that are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When the compound is a pure enantiomer, the stereochemistry at each chiral carbon can be specified by R or S. Resolved compounds with unknown absolute configuration can be specified as (+) or (-) based on the direction (dextrorotatory or levorotatory) in which they rotate plane-polarized light at the sodium D line wavelength. Some of the compounds described herein contain one or more asymmetric centers and thus can produce enantiomers, diastereomers, and other stereoisomers, whose asymmetric centers can be defined as (R)- or (S)- according to absolute stereochemistry. The chemical entities, pharmaceutical compositions, and methods of the present invention are intended to include all such possible stereoisomers, including racemic mixtures, optically pure forms, diastereomer mixtures, and intermediate mixtures. Optically active (R)- and (S)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. The optical activity of the compounds can be analyzed by any suitable method, including but not limited to chiral chromatography and optical rotation determination, and the degree of dominance of one stereoisomer relative to another can be determined.

[0129] In some embodiments, the compounds of this disclosure may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. The term "stereoisomer" can refer to a group of compounds having the same number and type of atoms and the same bonding between these atoms but different in three-dimensional structure. The term "stereoisomer" can refer to any member of this group of compounds. For example, a stereoisomer can be an enantiomer or a diastereomer. All stereoisomeric forms of the compounds of this disclosure and mixtures thereof, including racemic mixtures, are intended to form part of this disclosure.

[0130] When stereochemistry is not specified, certain molecules described herein include isomers, such as enantiomers and diastereomers, mixtures of enantiomers (including racemates), mixtures of diastereomers, and other mixtures thereof, provided they can be prepared by those skilled in the art through routine experiments. In some embodiments, a single enantiomer or diastereomer, i.e., the optically active form, can be obtained by asymmetric synthesis or by resolving a mixture of racemates or diastereomers. The resolution of a mixture of racemates or diastereomers (if possible) can be accomplished, for example, by conventional methods, such as crystallization in the presence of a resolving agent, or by chromatography using, for example, a chiral high-performance liquid chromatography (HPLC) column. Furthermore, a mixture of two enantiomers enriched in one of the two enantiomers can be purified by recrystallization and / or grinding to provide a further optically enriched form of the major enantiomer.

[0131] In some embodiments, the chiral center of this disclosure may have an S or R configuration as defined in the IUPAC 1974 recommendation.

[0132] As used herein, the term "group of a compound" refers to a structure derived from a parent compound by removing one or more atoms (e.g., hydrogen atoms). In one embodiment, "group of a compound" is a monovalent group derived by removing a hydrogen atom from a parent compound.

[0133] It should be understood that certain group naming conventions may include monovalent or divalent groups depending on the context. For example, when a substituent requires two connection points with the rest of the molecule, the substituent should be understood as a divalent group. For instance, substituents of alkyl groups that are identified as requiring two connection points include divalent groups, such as -CH2-, -CH2CH2-, -CH2CH(CH3)CH2-, etc. Other group naming conventions explicitly indicate that the group is a divalent group, such as "alkylene", "alkenyl", "arylene", etc.

[0134] In any case where a substituent is described as a divalent group (i.e., having two connection points with the rest of the molecule), it should be understood that, unless otherwise stated, the substituent may be connected in any configurational orientation.

[0135] A "tautomer" is a molecule in which a proton can transfer from one atom to another atom of the same molecule. In some embodiments, the compounds presented herein exist as tautomers. In cases where tautomerism is possible, a chemical equilibrium will exist between the tautomers. The exact proportions of the tautomers depend on several factors, including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibria include:

[0136]

[0137] "Stable compound" and "stable structure" can refer to compounds that are stable enough to be isolated from the reaction mixture with useful purity and formulated into effective therapeutic agents.

[0138] As used in this article, the phrases “parenteral administration” and “administered via parenteral route” refer to administration methods other than enteral and local administration, usually by injection, and including but not limited to intravenous, intramuscular, intra-arterial, intrathecal, intracapsular, intra-bursal, intraorbital, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, and intrasternal injections and infusions.

[0139] In this article, the phrase “pharmaceutically acceptable” is used to refer to compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues without causing excessive toxicity, irritation, allergic reactions, or other problems or complications, within the bounds of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio.

[0140] As used herein, the phrases “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” refer to pharmaceutically acceptable materials, compositions, or media, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials. Each carrier must be “acceptable,” meaning it is compatible with other components of the formulation and harmless to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and Soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic compatible substances used in pharmaceutical preparations.

[0141] As used in this disclosure, the term "carrier" can include carriers, excipients, and diluents, and can refer to materials, compositions, or media, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials, that participate in carrying or transporting an agent (such as one or more compounds of this disclosure or their pharmaceutically acceptable salts, solvates (e.g., hydrates), isomers (e.g., stereoisomers), and tautomers) from one organ or body part of a subject to another. The carrier should be selected based on the compatibility and release characteristics of the desired dosage form. Exemplary carrier materials may include, for example, excipients, binders, suspending agents, disintegrants, fillers, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, spray-dried dispersions, etc. See, for example, Hoover, John E. Remington’s Pharmaceutical Sciences , Mack Publishing Co., Easton, Pa. 1975. Exemplary carrier materials may also include, but are not limited to, any adjuvants, excipients, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0142] The terms “pharmaceutical acceptable” or “pharmacologically acceptable” can refer to a material that is biologically or otherwise undesirable—that can be administered to an individual without causing any substantial adverse biological effects or interacting in a harmful manner with any component of the composition therein.

[0143] "Pharmaceutical composition" can refer to a formulation of the compounds of this disclosure with a medium generally accepted in the art for delivering a bioactive compound to a subject (e.g., a mammal or a human). Such a medium can include all pharmaceutically acceptable carriers used therein.

[0144] The terms “subject,” “individual,” and “patient” are used interchangeably and refer to non-human mammals (e.g., non-human primates, dogs, horses, cats, pigs, cattle, ungulates, lagomorphs, etc.).

[0145] As used herein, the phrase “subjects in need” refers to subjects who have or are at risk of having a pathological condition as described below, which can be preventively or therapeutically treated with the compounds or salts described herein.

[0146] The terms “administer,” “administered,” “administers,” and “administering” are defined as the administration of a composition to a subject via routes known in the art, including but not limited to intravenous, intra-arterial, oral, parenteral, oral, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, transmucosal, or intraperitoneal administration. In some embodiments, an oral route of administration of the composition may be used.

[0147] The term "effective amount" or "therapeutic effective amount" refers to the amount of a compound or salt described herein sufficient to affect the intended application, including but not limited to the treatment of a disease as defined below. Therapeutic effective amounts can vary depending on the intended application (in vitro or in vivo), or the subject being treated and the disease condition (e.g., the subject's weight and age, the severity of the disease condition, the method of administration, etc.), which can be readily determined by one of ordinary skill in the art. This term may also apply to doses that can induce a specific response in target cells (e.g., reduced proliferation or downregulation of target protein activity). Specific doses can vary depending on the particular compound selected, the dosing regimen to be followed, whether it is administered in combination with other compounds, the time of administration, the tissue to which it is administered, and the physical delivery system carrying it.

[0148] As used herein, “treatment” or “treating” means a method for obtaining a beneficial or desired outcome (including, but not limited to, therapeutic and / or preventive benefits) for a disease, disorder, or medical condition. In some embodiments, treatment or treating involves administering to a subject a compound or composition disclosed herein. In some embodiments, therapeutic benefits may include relieving, reducing, or improving symptoms of a disease or condition; preventing additional symptoms; improving or preventing the underlying cause of symptoms; inhibiting a disease or condition, such as preventing its progression; alleviating a disease or condition; causing a remission of a disease or condition; relieving a condition caused by a disease or condition; or preventing symptoms of a disease or condition, whether preventive and / or therapeutic.

[0149] In some embodiments, the therapeutic benefit includes eradicating or improving the underlying impairment being treated. In some embodiments, the therapeutic benefit includes eradicating or improving one or more physiological symptoms associated with the underlying impairment, such as observing improvement in the subject, although the subject may still be troubled by the underlying impairment. In some embodiments, for preventative benefit, a compound or composition is administered to a subject at risk of developing a specific disease, or to a subject exhibiting one or more physiological symptoms of a disease, even if the disease may not yet be diagnosed. Treatment may include, for example, reducing, delaying, or alleviating the severity of one or more symptoms of a disease or condition, or it may include reducing the frequency with which a patient experiences symptoms of a disease, defect, impairment, or adverse condition. Herein, treatment can be used to refer to methods that result in some level of treatment or improvement of a disease or condition, and may encompass a range of outcomes for that purpose, including but not limited to complete prevention of the condition.

[0150] In some implementations, terms such as “prevent” or “preventing” related to a disease or disorder may refer to a compound that, in a statistical sample, reduces the incidence of a disorder or condition in a treated sample relative to an untreated control sample, or that, relative to an untreated control sample, delays the onset of one or more symptoms of a disorder or condition or reduces their severity.

[0151] In some implementations, the terms “disease” and “symptom” may be used interchangeably or may differ, meaning that a particular disease or symptom may not have a known causative agent (making the cause undetermined), and therefore it is not recognized as a disease but is only considered as an adverse condition or syndrome in which a clinician has identified a more or less specific set of symptoms.

[0152] Compounds of the disclosure

[0153] Generally, this disclosure provides compounds having a structure according to Formula I:

[0154] (I).

[0155] In some embodiments, such compounds exhibit affinity, inhibitory activity, or both for the canine JAK1 subtype and may be used to treat diseases and disorders mediated by the canine JAK1 subtype, such as canine atopic dermatitis. In some embodiments, compounds according to Formula I exhibit selectivity for the canine JAK1 subtype relative to other canine JAK subtypes (e.g., cJAK2, cJAK3, canine tyrosine kinase 2 (cTYK2), or combinations thereof). As described herein, this activity and selectivity may be desirable when providing companion animal therapeutics that have improved tolerability relative to known companion animal therapeutics for treating, for example, canine atopic dermatitis, and may offer the potential for once-daily dosing.

[0156] Therefore, in one aspect, a compound having a structure according to Formula I is provided.

[0157] (I)

[0158] Or a pharmaceutically acceptable salt or solvate thereof, wherein:

[0159] R1 is H or an optionally substituted C1-C3 alkyl group;

[0160] n = 0 or 1;

[0161] R2 is H or CH3;

[0162] R3 is a C1-C4 alkyl, phenyl,

[0163] ,or ,

[0164] in:

[0165] m is 0 or 1;

[0166] X is H, F, Cl, OCH3, or CH3;

[0167] R4 is H or CH3;

[0168] R5 is H, C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3, or 2-pyrimidinyl; or

[0169] R4 and R5, together with the contained N atom, form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally contains an additional heteroatom selected from N and O; or

[0170] R2 and R3 together with the contained N atom form a 6-membered ring, which is optionally converted into a C1-C4 alkyl group, C7 to C8 alkyl group, C9 alkyl group, C1 alkyl group, C2 alkyl group, C3 alkyl group, C4 alkyl group, C5 alkyl group, C6 alkyl group, C7 alkyl group, C9 alkyl group, C1 alkyl group, C2 alkyl group, C3 alkyl group, C4 alkyl group, C5 alkyl 12 Aryl or -C(O)-NR6R7 substitution,

[0171] in:

[0172] R6 is H or a C1-C4 alkyl group;

[0173] R7 is H, C1-C4 alkyl, or phenyl; or

[0174] R6 and R7 together with the contained N atom form a 6-membered ring, which may optionally be substituted with a C1-C4 alkyl group, and wherein the 6-membered ring optionally contains an additional N atom.

[0175] In some implementations, R1 is H.

[0176] In some implementations, R1 is CH3.

[0177] In some implementations, R2 is H.

[0178] In some implementations, R2 is H, and R3 is:

[0179] .

[0180] In some implementations, n is 1 and R1 is H.

[0181] In some implementations, R3 is:

[0182] .

[0183] In some implementations, m is 0. In some implementations, m is 1.

[0184] In some embodiments, X is H. In some embodiments, X is F or Cl. In some embodiments, X is OCH3. In some embodiments, X is CH3.

[0185] In some embodiments, R4 is H or CH3. In some embodiments, R4 is H. In some embodiments, R4 is CH3.

[0186] In some embodiments, R5 is H, C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3, or 2-pyrimidinyl.

[0187] In some embodiments, R4 and R5 together with the contained N atom form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally contains an additional heteroatom selected from N and O.

[0188] In some implementations, m is 0 and R4 and R5 are each H.

[0189] In some embodiments, the compounds of formula I are selected from:

[0190] and .

[0191] In some implementations, m is 1.

[0192] In some implementations, R4 and R5 are each CH3.

[0193] In some embodiments, the compound of formula I is:

[0194] .

[0195] In some embodiments, R1, R2, and R4 are H; and R5 is selected from C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3, and 2-pyrimidinyl.

[0196] In some embodiments, R1 is CH3; R2 and R4 are H; and R5 is selected from C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3 and 2-pyrimidinyl.

[0197] In some embodiments, R1 is H or CH3; R2 and R4 are H; and R5 is a C1-C4 alkyl group. In some embodiments, R1 is H or CH3; R2 and R4 are H; and R5 is a C3-C5 cycloalkyl group. In some embodiments, R1 is H or CH3; R2 and R4 are H; and R5 is CH2CH2OCH3. In some embodiments, R1 is H or CH3; R2 and R4 are H; and R5 is a 2-pyrimidinyl group.

[0198] In some embodiments, the compound of formula I is selected from...

[0199] .

[0200] In some embodiments, the compounds of formula I are selected from:

[0201] .

[0202] In some embodiments, the compounds of formula I are selected from:

[0203] .

[0204] In some embodiments, the compound of formula I is:

[0205] , ,or .

[0206] In some embodiments, the compound of formula I is:

[0207] , ,or .

[0208] In some embodiments, R2 and R3 together with the contained N atom form a 6-membered ring, which is optionally bounded by a C1-C4 alkyl group, C7 to C8 alkyl group, or C9-C9 alkyl group. 12 Aryl alkyl or -C(O)-NR6R7 substitution.

[0209] Therefore, in some embodiments, the compound of formula I has a structure according to formula II:

[0210] (II),

[0211] Where Z is either C or N.

[0212] In some implementations, Z is C. In some implementations, Z is N.

[0213] In some implementations, n is 1. In some implementations, n is 0.

[0214] In some implementations, R1 is H.

[0215] In some embodiments, R6 is H. In some embodiments, R6 is a C1-C4 alkyl group.

[0216] In some embodiments, R7 is H. In some embodiments, R7 is a C1-C4 alkyl group. In some embodiments, R7 is a phenyl group.

[0217] In some embodiments, R6 and R7 together with the contained N atom form a 6-membered ring, which may optionally be substituted with a C1-C4 alkyl group, and wherein the 6-membered ring optionally contains an additional N atom.

[0218] In some embodiments, the compound of formula II has a structure selected from the following:

[0219] .

[0220] In some embodiments, the compound of formula I has the following structure:

[0221] .

[0222] Pharmacological profile of compounds of the disclosure

[0223] Compounds of Formula I typically exhibit inhibitory activity against one or more Janus kinase (JAK) isoforms. The effects of compounds on JAK enzymes and / or animals can be determined or measured. Methods for determining JAK activity include those described in the examples of this disclosure, as well as those disclosed in WO1999 / 65908, WO1999 / 65909, WO2001 / 42246, WO2002 / 00661, WO2002 / 096909, WO2004 / 046112, or WO2007 / 012953, the contents of which are incorporated herein by reference.

[0224] In some embodiments, the compound of formula I has JAK1 activity, for example, through enzymatic binding to IC1. 50 The measured binding affinity is less than about 1 micromolar, for example less than about 500 nM, less than 250 nM, less than 100 nM, or less than 50 nM. In some embodiments, the compound of formula I is expressed in IC50. 50 The compounds of Formula I bind to JAK1 at a concentration of about 25 to about 100 nM, about 100 to about 250 nM, about 250 to about 500 nM, or about 500 to about 1000 nM. In some embodiments, the compounds of Formula I have human JAK1 activity, canine JAK1 activity, or both.

[0225] In some embodiments, compounds of formula I exhibit binding selectivity for JAK-1 relative to other enzymes (e.g., other JAK isoforms). In some embodiments, compounds of formula I exhibit selectivity for JAK-1 of about 5-fold or more, or about 10-fold or more, relative to JAK-2, for example, about 5 or about 10 to about 20, about 50 or about 100 times. In some embodiments, compounds of formula I are selective for human JAK-1. In some embodiments, compounds of formula I are selective for canine JAK-1. In some embodiments, compounds of formula I are selective for both human and canine JAK-1.

[0226] In some embodiments, the compound of Formula I is selective for inhibiting the function of JAK-1 (canine, human, or both). In some embodiments, the compound of Formula I has a selectivity of about 5 times or more, or about 10 times or more, for example about 5 or about 10 to about 20, about 50, or about 100 times for inhibiting JAK-1, relative to JAK-2 inhibition.

[0227] Preparation of compounds of the disclosure

[0228] The compounds of this disclosure can be prepared by methods known in the field of organic synthesis, as partially illustrated by the synthetic schemes described below, in examples, and in conjunction with the guidance provided herein. In the schemes described below and in the figures, it should be understood that protecting groups may be used on sensitive or reactive groups where necessary, based on general chemical principles or the guidance provided herein. Protecting groups can be manipulated according to standard methods of organic synthesis (TW Greene and PGM Wuts, “Protective Groups in Organic Synthesis,” 3rd edition, Wiley, New York, 1999). These groups can be removed at appropriate stages of the compound synthesis using methods readily apparent to those skilled in the art based on the detailed teachings provided herein. The selection of procedures, reaction conditions, and their sequence of execution should be consistent with this disclosure.

[0229] Typically, methods for preparing the compounds of this disclosure involve combinations of reactions and conditions. Schemes 1 and 2 illustrate representative, non-limiting alternative strategies for preparing compounds of formula (I). Various aspects of the preparation methods according to Schemes 1 and 2 are further discussed below. The reactions in the schemes can be carried out using chemical reactions according to standard synthetic chemical procedures and practices known in the art or described herein.

[0230] Referring to exemplary embodiment 1, in some embodiments, the compounds of this disclosure (e.g., compounds of formula I) can be prepared by amidation of a common intermediate carboxylic acid (4). Continuing with embodiment 1, in some embodiments, 4-chloro-7H-pyrrolo[2,3-d]pyrimidine 1 is reacted with piperazine 2 to form ester intermediate 3. In some embodiments, n is zero. In some embodiments, n is 1. In some embodiments, R1 is H. In some embodiments, R1 is a C1-C3 alkyl group, which may optionally be substituted. In some embodiments, R1 is methyl. In a particular embodiment, R1 is H and n is 0. The compounds of structure 2 may be commercially available or readily obtainable by known methods. Suitable conditions for alkylation of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine 1 with piperazine 2 include, but are not limited to, reactions in the presence of a nonnucleophilic amide base and in a polar solvent, typically at elevated temperatures. For example, the reaction can be conveniently carried out in n-butanol in the presence of diisopropylethylamine at about 90°C. Continuing with reference to Scheme 1, ester 3 can be saponified to provide acid 4. Suitable conditions include, but are not limited to, reaction with an alkali metal hydroxide (e.g., lithium hydroxide in aqueous methanol) in an aqueous alcohol solvent. Continuing with reference to Scheme 1, acid 4 is then coupled with the desired amine 5 to provide a compound of formula I. Those skilled in the art will recognize suitable coupling conditions. Suitable coupling conditions include, but are not limited to, those used for the formation of peptides in the presence of amine bases (e.g., pyridine, triethylamine, diisopropylethylamine, etc.) (e.g., HATU, TBTU, BOP, PyBOP, PyBROP, etc.). Amine component 5 can be commercially available or readily obtained by standard synthetic procedures (e.g., those further described below).

[0231] Option 1

[0232]

[0233] Scheme 2 provides an exemplary alternative synthetic scheme for compounds of Formula I. Referring to Scheme 2, in some embodiments, compounds of this disclosure (e.g., compounds of Formula I) can be prepared by alkylation of piperazine intermediate 6 with chloroamide 7. Chloroamide 7 can in turn be prepared from amine 5 and acyl chloride 8. Typically, piperazine intermediate 6 can be prepared as described above for Schemes 1 and intermediate 3 by reacting 4-chloro-7H-pyrrolo[2,3-d]pyrimidine 1 with piperazine. Suitable reaction conditions include, but are not limited to, the presence of an amine base (e.g., pyridine) and optionally a catalyst (e.g., pyridine). N,NThe reaction is carried out in an aprotic solvent in the presence of (-dimethylaminopyridine). In some embodiments, R1 is H and n is zero (i.e., 8 is chloroacetyl chloride). In some embodiments, alkylation with chloride 7 to 6 is carried out in a polar aprotic solvent (e.g., DMF) in the presence of an amine base (e.g., triethylamine), optionally in a catalytic or stoichiometric amount of sodium iodide or potassium iodide.

[0234] Option 2

[0235]

[0236] Continuing with reference to embodiments 1 and 2, in some embodiments, amine 5 is a primary amine. In some embodiments, amine 5 is a secondary amine. In some embodiments, amine 5 is a primary or secondary alkylamine, benzylamine, aromatic amine, or heterocyclic alkylamine. In some embodiments, R2 is H or CH3. In some embodiments, R3 is a C1-C4 alkyl or phenyl group. In some embodiments, R3 is:

[0237] ,or .

[0238] in:

[0239] m is 0 or 1;

[0240] X is H, F, Cl, OCH3, or CH3;

[0241] R4 is H or CH3;

[0242] R5 is H, C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3, or 2-pyrimidinyl; or

[0243] R4 and R5, together with the contained N atom, form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally contains an additional heteroatom selected from N and O; or

[0244] R2 and R3 together with the contained N atom form a 6-membered ring, which is optionally converted into a C1-C4 alkyl group, C7 to C8 alkyl group, C9 alkyl group, C1 alkyl group, C2 alkyl group, C3 alkyl group, C4 alkyl group, C5 alkyl group, C6 alkyl group, C7 alkyl group, C9 alkyl group, C1 alkyl group, C2 alkyl group, C3 alkyl group, C4 alkyl group, C5 alkyl 12 Aryl or -C(O)-NR6R7 substitution,

[0245] in:

[0246] R6 is H or a C1-C4 alkyl group;

[0247] R7 is H, C1-C4 alkyl, or phenyl; or

[0248] R6 and R7 together with the contained N atom form a 6-membered ring, which may optionally be substituted with a C1-C4 alkyl group, and wherein the 6-membered ring optionally contains an additional N atom.

[0249] In some implementations, R2 is H and R3 is

[0250] ,

[0251] Where m, X, R4 and R5 are each defined as described above.

[0252] In some implementations, m is 1.

[0253] In some implementations, m is 1, X is H, and amine 5 is selected from:

[0254] .

[0255] Such amines are usually commercially available or can be readily prepared using methods known to those skilled in the art.

[0256] In some embodiments, m is 0. Therefore, in some embodiments, amine 5 has structure 9:

[0257] ,

[0258] X, R4, and R5 are each as described above. Such amines are commercially available or can be readily prepared according to known methods. For example, sulfonamide 9 can be conveniently prepared from the corresponding nitrobenzenesulfonyl chloride 10 according to Scheme 3. Referring to Scheme 3, generally, the commercially available nitrobenzenesulfonyl chloride 10 is reacted with amine 11 in the presence of a suitable base (e.g., pyridine or a non-nucleophilic amine) to form sulfonamide 12. Such sulfonamides can be readily reduced to aminosulfonamide 9, for example, by hydrogenation over a suitable catalyst or by reduction with iron, zinc, etc., under acidic conditions.

[0259] Option 3

[0260]

[0261] In some embodiments, sulfonamide 9 is selected from:

[0262] ,

[0263] in:

[0264] X is H, F, Cl, OCH3, or CH3; and

[0265] R5 is a C1-C4 alkyl, C3-C5 cycloalkyl, CH2CH2OCH3, or 2-pyrimidinyl.

[0266] In some embodiments, sulfonamide 9 is selected from:

[0267] .

[0268] In some embodiments, sulfonamide 9 is selected from:

[0269] .

[0270] In some implementations, amine 5 is:

[0271] .

[0272] In some embodiments, the compound of formula I has a structure according to formula II:

[0273] (II),

[0274] Where Z is C or N, and R1, R6, R7, and n are each as described above. Therefore, in some embodiments, amine 5 has a structure according to formula III:

[0275] (III).

[0276] In some embodiments, Z is C. Such amines can be prepared by coupling N-protected piperidine-4-carboxylic acid with amine HNR6R7 according to methods known to those skilled in the art.

[0277] In some implementations, amine 5 is:

[0278] .

[0279] In some implementations, amine 5 is:

[0280] .

[0281] In some embodiments, Z is N. Such amines can be prepared by coupling a mono-N-protected piperazine with the amine HNR6R7 and a phosgene equivalent according to methods known to those skilled in the art.

[0282] In some implementations, amine 5 is:

[0283] .

[0284] In some embodiments, the compound of formula I has a structure according to formula IV:

[0285] (IV),

[0286] Where Ar is a phenyl or a substituted phenyl, and L is an integer from 1 to 6. Therefore, in some embodiments, amine 5 has the following structure:

[0287] 。

[0288] Such amines can be prepared by the coupling reaction between an aryl halide and a piperidinyl olefin according to Exemplary Scheme 4. Referring to Scheme 4, an aryl bromide is coupled with a suitable piperidinyl olefin (13, 14 or 15) under palladium catalysis (e.g., Heck conditions). Those skilled in the art will recognize that piperidine olefin 13 will provide amine 5 wherein L is 1; piperidine olefin 14 will provide amine 5 wherein L is 2; and piperidine olefin 15 wherein p is 1, 2, 3 or 4 will provide amines 5 wherein L is 3, 4, 5 or 6, respectively. Those skilled in the art will also recognize that suitable protecting groups (e.g., BOC) can be used on the nitrogen of the corresponding piperidine olefins.

[0289] Scheme 4

[0290]

[0291] In some embodiments, Ar is phenyl and L is 1. Thus, in some embodiments, amine 5 is:

[0292] 。

[0293] Isotopes and Isotopically Labeled Compounds

[0294] The compounds described herein may occur in their natural isotopic abundances, or one or more atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. All isotopic variants of the compounds of the present disclosure, whether radioactive or not, are included within the scope of the present disclosure. Thus, reference to an element is intended to include all isotopes of that element. For example, if the R group is defined to include hydrogen or H, it also includes isotopes of hydrogen. For example, hydrogen has three naturally occurring isotopes, designated as 1 H (protium), 2 H (deuterium), and 3 H (tritium). Protium is the most abundant hydrogen isotope in nature. Enrichment of deuterium can provide certain therapeutic advantages, such as increased in vivo half-life and / or exposure, or can provide compounds useful for studying drug elimination and metabolic pathways in vivo. Isotopically enriched compounds can be prepared by conventional techniques well known to those skilled in the art.

[0295] The compounds described herein also include all pharmaceutically acceptable isotopically labeled compounds. An "isotopically labeled" or "radiolabeled" compound can be one in which one or more atoms are replaced or substituted with atoms having an atomic mass or mass number different from those normally found in nature (i.e., naturally occurring). For example, in some embodiments, in the compounds described herein, hydrogen atoms are replaced or substituted with one or more deuterium or tritium atoms.

[0296] Certain isotope-labeled compounds disclosed herein, such as those doped with radioactive isotopes, can be used for drug and / or substrate tissue distribution studies. Radioactive isotope tritium (i.e., 3 H) and carbon-14 (i.e., ... 14 C) It is particularly suitable for this purpose due to its ease of incorporation and convenient detection methods. Using heavier isotopes (e.g., deuterium, i.e., 2 H) substitution may offer certain therapeutic advantages due to increased metabolic stability, such as increased in vivo half-life or reduced dose requirements, and may therefore be preferred in some cases. In some embodiments, the compound contains at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of this disclosure may be replaced or substituted with deuterium. In some embodiments, the compound contains two or more deuterium atoms. In some embodiments, the compound contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Suitable isotopes that may be incorporated into the compounds described herein include, but are not limited to, those found in the present disclosure. 2 H (also written as D for deuterium) 3 H (also written as T for tritium) 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 18 F, 35 S, 36 Cl、 82 Br、 75 Br、 76 Br、 77 Br、 123 I, 124 I, 125 I and 131 I. Using positron emission of isotopes (e.g.) 11 C 18 F, 15 O and 13 N) substitution can be used in positron emission tomography (PET) studies.

[0297] Isotopically labeled versions of the compounds disclosed herein can generally be prepared by replacing non-isotopically labeled reagents with appropriate isotopically labeled reagents by following a procedure similar to that disclosed in the schemes and / or examples herein.

[0298] Isomers

[0299] In some embodiments, the compounds of this disclosure may be enriched to primarily provide one enantiomer of the compounds described herein. The enantiomer-enriched mixture may contain, for example, at least 60 mol% of one enantiomer, or at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, at least 99.5, or even 100 mol%. In some embodiments, the compounds described herein enriched with one enantiomer may be substantially free of the other enantiomer, where substantially free may mean, for example, that in the compound mixture, the substance comprises less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% compared to the amount of the other enantiomer. For example, if the compound mixture contains 98 grams of the first enantiomer and 2 grams of the second enantiomer, it can be said to contain 98 mol% of the first enantiomer and only 2 mol% of the second enantiomer.

[0300] In some embodiments, the compounds of this disclosure may be enriched to primarily provide one diastereomer of the compounds disclosed herein. The diastereomer-enriched mixture may contain, for example, at least 60 mol% of one diastereomer, or at least 75, at least 80, at least 85, at least 90, at least 95, at least 96, at least 97, at least 98, at least 99, at least 99.5, or even 100 mol%. In some embodiments, the compounds enriched with one diastereomer described herein may be substantially free of other diastereomers, where substantially free may mean, for example, that the substance comprises less than 10%, or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1% of the amount of other diastereomers in the compound mixture.

[0301] A mixture of diastereomers can be separated into their respective diastereomers based on their physicochemical differences using methods well known to those skilled in the art (e.g., by chromatography and / or fractional crystallization). Enantiomers can be separated by reacting the enantiomer mixture with a suitable optically active compound (e.g., a chiral auxiliary, such as a chiral alcohol or Mosher's acyl chloride), separating the diastereomers, and converting (e.g., hydrolyzing) each diastereomer to its corresponding pure enantiomer. Enantiomers can also be separated using a chiral HPLC column. Furthermore, some compounds of this disclosure may be in the form of transisomers or rotational isomers and are considered part of this disclosure.

[0302] Metabolites

[0303] The disclosure herein is also intended to cover in vivo metabolites of the disclosed compounds. Such products may be generated, for example, by oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound, primarily due to enzymatic processes. Therefore, this disclosure may include compounds produced by methods comprising administering a compound of this disclosure to a subject (e.g., a mammal) for a period sufficient to produce its metabolites. Such products are typically determined by administering a radiolabeled compound of this disclosure to a subject (e.g., a rat, mouse, guinea pig, monkey, or human) at a detectable dose, allowing sufficient time for metabolism, and isolating its metabolites from urine, blood, or other biological samples.

[0304] Salts and solvates

[0305] This disclosure also provides pharmaceutically acceptable salts, solvates (e.g., hydrates), and combinations thereof of any compounds disclosed herein. The terms “salt,” “hydrate,” “solvent,” etc., are intended to equally apply to salts, hydrates, or solvates of enantiomers, diastereomers, isomers, stereoisomers, rotational isomers, tautomers, positional isomers, or racemates of the disclosed compounds.

[0306] The term "salt" or "pharmaceutically acceptable salt" refers to a salt derived from a variety of organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can form with both inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. Pharmaceutically acceptable base addition salts can form with both inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, etc., specifically such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium, potassium, sodium, calcium, and magnesium salts. In some implementations, "pharmaceutically acceptable salts" may include, for example, water-soluble and water-insoluble salts such as acetates, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonates, benzoates, bicarbonates, bisulfates, tartrates, borates, bromides, butyrates, calcium salts, calcium ethylenediaminetetraacetate, camphorsulfonates, carbonates, chlorides, citrates, clavulanates, dihydrochlorides, ethylenediaminetetraacetate, ethanedisulfonates, estolates, ethanesulfonates, fiunarates, glucono-p-ethylhexanoate, gluconate, glutamate, glycolyllarsanilate, hexafluorophosphates, hexylresorcinol salts, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthyl carbamate, iodides, hydroxyl... Sethionate, lactate, lacturonate, laurate, magnesium salt, malate, maleate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, mucilage, naphthalene sulfonate, nitrate, N-methylglucosamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, dihydroxynaphthoate, 1,1-methylene-bis-2-hydroxy-3-naphthoate, einbonate, pantothenate, phosphate / bisphosphonate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, basic acetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, theochloroate, toluenesulfonate, triethiodide, and valerate.

[0307] Suitable anionic salt forms include, but are not limited to, acetates, benzylsulfonates, benzoates, benzylates, bicarbonates, tartrates, bromides, calcium ethylenediaminetetraacetate, camphor sulfonates, carbonates, chlorides, citrates, dihydrochlorides, ethylenediaminetetraacetate, ethanedisulfonates, etolates, ethanesulfonates, fumarates, glucono-p-glucose, gluconate, glutamate, p-hydroxyacetaminophenarsine, hexylresorcinol, hyaluronic acid, hydrobromide, and hydrochloric acid. Salts, hydroxynaphthyl carboxylates, iodides, hydroxyethanesulfonates, lactates, lacturonates, malates, maleates, mandelates, methanesulfonates, methyl bromides, methyl nitrates, methyl sulfates, mucilages, naphthalenesulfonates, nitrates, bis(hydroxynaphthyl) salts (emborates), pantothenates, phosphates and diphosphates, polygalacturonates, salicylates and disalicylates, stearates, basic acetates, succinates, sulfates, tannins, tartrates, theochloroate, toluenesulfonates, triethyl iodide, valerates, etc.

[0308] Suitable cation salt forms include, but are not limited to, aluminum salts, benzyl sulfadiazine salts, calcium salts, ethylenediamine salts, lysine salts, magnesium salts, meglumine salts, potassium salts, procaine salts, sodium salts, tromethamine salts, and zinc salts.

[0309] In some embodiments, the salt is selected from acetate, ascorbate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, hydrogen sulfate / sulfate, borate, camphorsulfonate, citrate, ethanedisulfonate, α-ketoglutarate, ethanesulfonate, formate, fumarate, glucohepanoate, glucuronate, glyceryl phosphate, hexafluorophosphate, hymenate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, hydroxyethanesulfonate, lactate, malate, maleate, malonate, methanesulfonate, methyl sulfate, naphthylformate, 2-naphthylsulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, dihydroxynaphthyl, phosphate / hydrogen phosphate / dihydrogen phosphate, sucrose, stearate, succinate, tartrate, toluenesulfonate, and trifluoroacetate.

[0310] The compounds disclosed herein also include crystalline and amorphous forms of these compounds, pharmaceutically acceptable salts, and active metabolites of these compounds having the same type of activity, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolventized polymorphs (including anhydrous forms), conformational polymorphs, and amorphous forms of the compounds, as well as mixtures thereof.

[0311] The compounds of this disclosure, including their stereoisomers and tautomers, and salts of any of them, can exist as solvates. Typically, crystallization produces solvates of the compounds of this disclosure. As used herein, the term "solvate" can refer to an aggregate comprising one or more molecules of the compound of this disclosure and one or more solvent molecules. The solvent can be water, in which case the solvate can be a hydrate. Alternatively, the solvent can be an organic solvent. Therefore, the compounds and salts of this disclosure can exist as hydrates, including monohydrates, dihydrates, hemihydrates, sesquihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of this disclosure can be true solvates, while in other cases, the compounds of this disclosure may contain only exogenous water or a mixture of water and some exogenous solvent.

[0312] Pharmaceutical formulations and compositions

[0313] The compounds, salts, solvates, isomers, etc., disclosed herein can be administered to a subject, alone or in pharmaceutical compositions, wherein the compounds, salts, solvates, isomers, etc., disclosed herein are mixed with one or more biologically suitable and pharmaceutically acceptable carriers or excipients. The compositions can be in various forms, including but not limited to oral formulations, injectable formulations, suppository formulations, and topical, skin, or subcutaneous formulations. The selection of appropriate formulations of the compounds provided herein can be based, for example, on the physicochemical properties of the compound (and any other optional active pharmaceutical agent to be administered), the type of animal to be treated, the condition of the animal to be treated, and cost.

[0314] Pharmaceutical compositions may comprise at least one compound or salt described herein, and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents, and / or thickeners. Specific components and the relative amounts of each component will vary depending on, for example, the intended route of administration. The compositions may be formulated to contain a single daily dose or an appropriate amount of a daily dose in a dosage unit (e.g., a single tablet, a single capsule, an appropriate volume of liquid / ointment, etc.). The amount of compounds of the above-provided formula contained in a given composition for use in animal and human health can vary widely. The compounds are typically contained in the composition in amounts likely to induce the desired effect. Compositions to be administered to a subject or patient are typically in the form of one or more dosage units, wherein, for example, a tablet may be a single dosage unit, while a container for one or more compounds of this disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers (in aerosol form) may contain multiple dosage units.

[0315] Pharmaceutical compositions used according to this disclosure can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers, said carriers containing excipients and adjuvants that facilitate the processing of the active compound into a pharmaceutically usable preparation. Appropriate formulations depend on the chosen route of administration. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are therefore included in this invention. Such excipients and carriers are described, for example, in “Remingtons Pharmaceutical Sciences”, Mack Publishing Co., New Jersey (1991). Generally, pharmaceutical compositions of the disclosed compounds can be prepared in ways known per se, such as by conventional mixing, dissolving, granulation, pelleting, grinding, emulsification, encapsulation, embedding, or lyophilization processes. Techniques for formulation can be found in references well known to those skilled in the art, such as “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, PA, latest edition.

[0316] Oral dosage forms

[0317] The compositions described herein are formulated for oral administration. When intended for oral administration, the pharmaceutical compositions of this disclosure are generally in solid or liquid form, wherein semi-solid, semi-liquid, suspension, and gel forms may be included in forms regarded herein as solid or liquid.

[0318] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) a filler or extender, such as starch, lactose, sucrose, glucose, mannitol, and silicate; b) a binder, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; c) a humectant, such as glycerin; d) a disintegrant, such as agar, calcium carbonate, potato or cassava starch, alginic acid, certain silicates, and sodium carbonate; e) a solution retarder, such as paraffin; f) an absorption enhancer, such as a quaternary ammonium compound; g) a wetting agent, such as cetyl alcohol and glyceryl monostearate; h) an absorbent, such as kaolin and bentonite; and i) a lubricant, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer. When the pharmaceutical composition is in the form of capsules (e.g., gelatin capsules), it may also contain a liquid carrier such as polyethylene glycol or oil, in addition to the materials disclosed herein.

[0319] Similar solid compositions can also be used as fillers in soft-filled and hard-filled gelatin capsules, which use excipients such as lactose or milk sugar and high molecular weight polyethylene glycol. Solid dosage forms such as tablets, sugar-coated pills, capsules, pellets, and granules can be prepared using coatings and shells (e.g., enteric coatings and other coatings well known in the pharmaceutical formulation field). They may optionally contain light-blocking agents and may also be compositions that release (multiple) active ingredients only or preferentially in a portion of the intestine, optionally in a delayed manner. Examples of encapsulation compositions that can be used include polymers and waxes. In some embodiments, capsules (hard or soft capsules) comprising compounds according to the formulas described herein are provided, such as gelatin capsules. In some embodiments, hard gelatin capsules may comprise compounds according to the formula described herein, which are mixed with an inert solid diluent (e.g., starch, powdered cellulose (e.g., crystalline or microcrystalline cellulose), sugar (e.g., fructose, mannitol, or sucrose), cereal flour, calcium carbonate, calcium phosphate, and / or kaolin) and encapsulated within a capsule. In some embodiments, soft or liquid gelatin capsules may comprise compounds according to the formula described herein, which are mixed with water or a solvent (e.g., propylene glycol, polyethylene glycol (PEG), and / or ethanol), or with an oil medium (e.g., peanut oil, liquid paraffin, or olive oil), and encapsulated within a capsule. In some embodiments, such capsules are microcapsules.

[0320] In some embodiments, compositions in tablet form are provided, comprising a compound according to the formula described herein and one or more pharmaceutically acceptable excipients. Tablets are typically prepared by direct compression (e.g., wet or dry granulation of the ingredients). Excipients suitable for tablet preparation are generally known and include, for example, inert diluents (e.g., starch, lactose, mannitol, powdered sugar, powdered cellulose derivatives, kaolin, calcium carbonate, sodium carbonate, lactose, calcium phosphate, calcium sulfate, sodium phosphate and / or inorganic salts such as sodium chloride); granulating agents and disintegrants (e.g., starch such as corn or potato starch, clay, cellulose, methylcellulose, carboxymethylcellulose, alginic acid and / or seaweed extract, agar, bentonite, wood cellulose, powdered natural sponge, cation exchange resin, guar gum, citrus pulp and / or sodium lauryl sulfate); binders (e.g., starch, gelatin, sugars (e.g., lactose, fructose, glucose, etc.), natural or synthetic gums such as gum arabic, alginate, methylcellulose, polyvinylpyrrolidone, etc.), polyethylene glycol, ethylcellulose and / or waxes; and lubricants (e.g., magnesium stearate, calcium stearate, stearic acid, hydrogenated vegetable oil and / or talc). Tablets may be uncoated or coated using known techniques (which can be used to delay disintegration and absorption in the gastrointestinal tract, thereby providing sustained action over a longer period). For example, in some embodiments, time-delaying materials such as glyceryl monostearate or glyceryl distearate may be used. Tablets may also optionally be coated using techniques described in U.S. Patent Nos. 4,256,108 (Theeuwes); 4,166,452 (Generales Jr.); and 4,265,874 (Bonsen et al.), which relate to the preparation of controlled-release osmotic therapeutic tablets and are incorporated herein by reference. Tablets are an example of tablets, typically formulated as small, hard tablets that dissolve slowly when placed under the tongue. Tablets may optionally be coated, for example with sugar as a flavoring and sealing agent, or with film-forming protectants to modify the dissolution properties of the tablet.

[0321] The pharmaceutical compositions of this disclosure may be in liquid form, such as elixirs, syrups, solutions, emulsions, or suspensions. Liquids may be intended for oral administration or delivery by injection (as two examples). When intended for oral administration, in addition to containing one or more compounds of this disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers, the pharmaceutical compositions of this disclosure typically also contain one or more sweeteners, preservatives, dyes / colorants, and flavor enhancers.

[0322] In some embodiments, compositions comprising compounds according to the formulas described herein are provided in the form of emulsions, such as oil-in-water or water-in-oil emulsions. The oil phase of the emulsion may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture of any such oils. In some embodiments, the oil phase consists of unsaturated polysaccharide-based glycerides, triglycerides (e.g., medium-chain triglycerides, such as C8-C...). 10 The emulsion is formed from caprylic / capric triglycerides or combinations thereof. The aqueous phase may contain water or glycol derivatives (e.g., propylene glycol, ethylene glycol ethers, polyethylene glycol, or glycerol). Specific examples include, but are not limited to, propylene glycol, diethylene glycol monoethyl ether, dipropylene glycol monomethyl ether, and mixtures thereof. Suitable emulsifiers may include, for example, naturally occurring phospholipids, such as soybean or lecithin; and esters or metaesters derived from fatty acids and hexitan anhydrides, such as sorbitan monooleate; and condensation products of the aforementioned metaesters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also optionally contain sweeteners, bittering agents, flavoring agents, and / or preservatives. In one embodiment, the emulsion is in the form of a microemulsion (consisting of a stable dispersion of aqueous phase droplets in an oil phase or oil phase droplets in an aqueous phase). Microemulsions are quaternary systems comprising an aqueous phase, an oil phase, a surfactant, and a co-surfactant. They are translucent and isotropic liquids. These droplets are less than 200 nm in size (in contrast, emulsion droplets range from approximately 1,000 nm to 100,000 nm). In some embodiments, the oil phase will comprise the microemulsion in the range of % v / v selected from: about 2% to about 15% v / v; about 7% to about 10% v / v; and about 8% to about 9% v / v. Typically, the aqueous phase will comprise the microemulsion in the range of about 1% to about 4% v / v. The interfacial film is composed of alternating surfactant (SA) and co-surfactant (Co-SA) molecules, which enable the microemulsion to form spontaneously by reducing interfacial tension. Surfactants used in the microemulsion include diethylene glycol monoethyl ether, dipropylene glycol monomethyl ether, polyethylene glycol-modified C8-C10 glycerol esters, or polyglycerol-6 dioleate. In addition to these surfactants, co-surfactants include short-chain alcohols such as ethanol and propanol. Some compounds are common to the three components discussed above (i.e., the aqueous phase, surfactant, and co-surfactant). However, using different compounds for various components of the same formulation is entirely within the skill level of those skilled in the art. In one embodiment, the ratio of co-surfactant to surfactant will be about 1 / 7 to about 1 / 2. In another embodiment, the microemulsion will contain about 25 to about 75% v / v of surfactant and about 10 to about 55% v / v of co-surfactant.

[0323] In some embodiments, compositions comprising compounds according to formulas as described herein are provided, which are in the form of suspensions and typically comprise compounds dispersed in a liquid (optionally together with other ingredients). The compounds are typically in the form of dispersible powders or granules; dispersible powders and granules suitable for preparing suspensions typically provide an active ingredient mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents can be illustrated by those already mentioned above. Other excipients may also be present, such as sweeteners, bittering agents, flavoring agents, and coloring agents. The liquid may be oily or aqueous. Oily suspensions can be formulated by suspending the compound in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (e.g., liquid paraffin). Oily suspensions may contain thickeners such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners (e.g., sucrose, saccharin, or aspartame), bittering agents, and flavoring agents may be added to provide a palatable oral formulation. These compositions can be preserved by adding antioxidants (such as ascorbic acid) or other known preservatives. Aqueous suspensions comprise compounds mixed with excipients suitable for preparing aqueous suspensions. For example, such aqueous suspensions may contain excipients as suspending agents, such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; dispersants or wetting agents may be naturally occurring phospholipids, such as lecithin, or condensation products of alkyl esters and fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide and long-chain aliphatic alcohols, such as heptadecaethyleneoxycetyl alcohol, or condensation products of ethylene oxide and esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or condensation products of ethylene oxide and esters derived from fatty acids and hexitol anhydrides, such as polyvinyl dehydrated sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives, such as ethylparaben or n-propylparaben, one or more colorants, one or more flavoring agents, and one or more sweeteners and / or bittering agents, such as those described above. Aqueous suspensions may contain, for example, fruit juice (such as apple juice or orange juice).

[0324] In some embodiments, compositions comprising compounds according to the formulas described herein are provided in the form of syrups or elixirs. Syrups and elixirs may be formulated with sweeteners (e.g., glycerin, propylene glycol, sorbitol, or sucrose). Such formulations may also contain modifiers, preservatives, flavoring agents, and / or coloring agents.

[0325] In some embodiments, compositions comprising compounds according to the formulas described herein are in paste form. Examples of embodiments in paste form include, but are not limited to, those described in U.S. Patent Nos. 6,787,342 (Chen), 7,001,889 (Freehauf et al.), and 7,563,773 (Freehauf), each of which is incorporated herein by reference in its entirety. In addition to the compounds of the present invention, the paste may also contain, for example, fumed silica; viscosity modifiers (e.g., selected from PEG 200, PEG 300, PEG 400, PEG 600, monoethanolamine, triethanolamine, glycerol, propylene glycol, polyoxyethylene (20) sorbitan monooleate (polysorbate 80 or Tween 80) and poloxamer (e.g., Pluronic L 81)); carriers (e.g., hydrophilic carriers selected from triacetin, monoglycerides, diglycerides and triglycerides); optionally, absorbents (e.g., selected from magnesium carbonate, calcium carbonate, starch, cellulose and their derivatives); and optionally, colorants (e.g., selected from titanium dioxide, iron oxide and FD&C Blue #1 aluminum lake), stabilizers, surfactants and / or preservatives.

[0326] Sustained-release formulations can also be prepared. Examples of sustained-release formulations may include a semi-permeable matrix of a solid hydrophobic polymer containing the compound or its salts, which may be in the form of a molded article (e.g., a film or microcapsule). Examples of sustained-release matrices may include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactide, copolymers of L-glutamic acid and γ-ethyl-L-glutamic acid, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT™ (i.e., injectable microspheres composed of lactic acid-glycolic acid copolymers and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.

[0327] Injectable forms

[0328] The compositions described herein can be formulated for administration as injections. Compositions intended for administration by injection may contain one or more of surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers, and isotonic agents. Non-limiting examples of injectable formulations may include sterile suspensions, solutions, or emulsions in oily or aqueous media. Suitable oily media may include, but are not limited to, lipophilic solvents or media such as fatty oils or synthetic fatty acid esters, or liposomes. Aqueous injectable suspensions may contain substances that increase the viscosity of the suspension. Suspensions may also contain suitable stabilizers. Injections can be formulated for bolus injection or continuous infusion. Alternatively, the compositions described herein may be lyophilized or in powder form prior to use for reconstitution with a suitable media (e.g., sterile pyrogen-free water).

[0329] For parenteral administration, the compound or salt may be formulated in a unit dose injectable form (e.g., using a solution, suspension, or emulsion) in combination with a pharmaceutically acceptable parenteral medium. Such a medium may be inherently non-toxic and non-therapeutic. The medium may be water, saline, Ringer's solution, dextran solution, and 5% human serum albumin. Non-aqueous mediums, such as fixed oils and ethyl oleate, may also be used. Liposomes may be used as carriers. The medium may contain small amounts of additives, such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives).

[0330] The liquid pharmaceutical compositions disclosed herein, whether solutions, suspensions, or other similar forms, may include one or more of the following adjuvants: sterile diluents, such as water for injection, saline solution, physiological saline, Ringer's solution, isotonic sodium chloride, fixed oils (e.g., synthetic monoglycerides or diglycerides that can serve as solvents or suspension media), polyethylene glycol, glycerol, propylene glycol, or other solvents; antimicrobial agents, such as benzyl alcohol or methylparaben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetates, citrates, or phosphates; and agents for adjusting tonicity, such as sodium chloride or dextran. Parenteral formulations may be packaged in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. In some embodiments, the adjuvant is physiological saline. In some embodiments, the injectable pharmaceutical composition is sterile.

[0331] Other dosage forms

[0332] The pharmaceutical compositions disclosed herein may be intended for topical application, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel matrix. For example, the matrix may comprise one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents (such as water and alcohol), and emulsifiers and stabilizers. Thickeners may be present in pharmaceutical compositions intended for topical application.

[0333] In some embodiments, compositions suitable for topical, dermal, and / or subcutaneous formulations are provided, comprising compounds according to multiple formulas described herein. Topical, dermal, and subcutaneous formulations may include, for example, emulsions, creams, ointments, gels, pastes, powders, patches, shampoos, pour-on formulations, ready-to-use formulations, spray formulations, and spot-on formulations. Such formulations may be, for example, concentrated solutions, suspensions, microemulsions, or emulsions. In some embodiments, topical application may cause the distribution of multiple active compounds through the glands (e.g., sebaceous glands) of an animal, and / or cause the multiple active compounds to achieve a systemic effect (plasma concentration), and / or allow distribution throughout the coat. Certain suitable formulations for topical, dermal, and / or subcutaneous application include, but are not limited to, those disclosed in U.S. Patent No. 6,395,765 (Etchegaray), which is incorporated herein by reference in its entirety.

[0334] Spot-applied compositions are typically applied to localized areas, which are distinct from the entire body of the animal. Spot-applied compositions are usually applied by applying the composition to a specific location on the animal (e.g., between the shoulders). Another embodiment of a localized area is a strip-shaped area, such as a strip running from the animal's head to its tail. Such compositions can be applied by, for example, pipetting, squeezing, or dripping.

[0335] The carrier may be a liquid carrier medium as described in U.S. Patent No. 6,426,333, which is incorporated herein by reference in its entirety. For example, in one embodiment, the dot application formulation comprises a solvent and a co-solvent, wherein the solvent is selected from acetone, acetonitrile, benzyl alcohol, diethylene glycol butyl ether, dimethylacetamide, dimethylformamide, dipropylene glycol n-butyl ether, ethanol, isopropanol, methanol, ethylene glycol monoethyl ether, ethylene glycol monomethyl ether, monomethylacetamide, dipropylene glycol monomethyl ether, liquid polyoxyethylene glycol, propylene glycol, 2-pyrrolidone (e.g., N-methylpyrrolidone), diethylene glycol monoethyl ether, ethylene glycol, diethyl phthalate, fatty acid esters (e.g., diethyl adipate or diisobutyl adipate), and mixtures of at least two of these solvents; the co-solvent is selected from anhydrous ethanol, isopropanol, or methanol. The liquid carrier medium may optionally contain a crystallization inhibitor selected from anionic surfactants, cationic surfactants, nonionic surfactants, amine salts, amphoteric surfactants, or copolymers of polyvinylpyrrolidone, polyvinyl alcohol, vinyl acetate and vinylpyrrolidone, polyethylene glycol, benzyl alcohol, mannitol, glycerol, sorbitol, polyoxyethylene dehydrated sorbitol ester; lecithin, sodium carboxymethyl cellulose and acrylic acid derivatives, or mixtures of these crystallization inhibitors.

[0336] Pouring compositions are typically used by pouring the composition along the animal's dorsal line (e.g., from the neck to the tail). Pouring formulations are advantageously oily and typically contain a diluent or carrier, and if the active ingredient is insoluble in the diluent, also contain a solvent (e.g., an organic solvent). Certain non-limiting pouring compositions are disclosed, for example, in U.S. Patent No. 6,010,710 to Etchegaray and U.S. Patent No. 8,097,266 to Gogolewski et al., which are incorporated herein by reference in their entirety.

[0337] Spray compositions are typically used by spraying the composition along the animal's back line (e.g., from the neck to the tail). Each of these compositions can involve application, such as a concentrated solution, suspension, microemulsion, or emulsion.

[0338] Such topical compositions (e.g., dotting, spraying, and pouring compositions) may typically contain compounds as provided herein, in combination with one or more diluents / media and / or one or more solvents. Diluents / media include, but are not limited to, vegetable oils (e.g., soybean oil, peanut oil, castor oil, corn oil, cottonseed oil, olive oil, grapeseed oil, sunflower oil, etc.); mineral oils (e.g., petrolatum, paraffin, silicone, etc.); aliphatic or cyclic hydrocarbons; medium-chain hydrocarbons (e.g., C8 to C9). 12Triglycerides and combinations thereof. Solvents (e.g., organic solvents) that may be added in some embodiments include, but are not limited to, acetyl tributyl citrate, fatty acid esters (e.g., dimethyl adipate, diisobutyl adipate), acetone, acetonitrile, benzyl alcohol, diethylene glycol butyl ether, dimethylacetamide, dimethylformamide, dipropylene glycol n-butyl ether, ethanol, isopropanol, methanol, ethylene glycol monoethyl ether, ethylene glycol monomethyl ether, monomethylacetamide, dipropylene glycol monomethyl ether, liquid polyoxyethylene glycol, propylene glycol, 2-pyrrolidone (e.g., N-methylpyrrolidone), diethylene glycol monoethyl ether, ethylene glycol, and diethyl phthalate, and mixtures of two or more thereof. In some embodiments, the composition contains emollients and / or spreaders and / or film-forming agents. Emollients and / or spreaders and / or film-forming agents may be selected, for example, from: polyvinylpyrrolidone, polyvinyl alcohol, copolymers of vinyl acetate and vinylpyrrolidone, polyethylene glycol, benzyl alcohol, mannitol, glycerin, sorbitol, polyoxyethylene dehydrated sorbitol ester; lecithin, sodium carboxymethyl cellulose, silicone oil, polydiorganosiloxane oil (e.g., polydimethylsiloxane (PDMS) oil), such as those containing silanol functional groups, or 45V2 oil; anionic surfactants, such as basic stearates, sodium stearate, potassium stearate, or ammonium stearate; calcium stearate, triethanolamine stearate; sodium rosinate; alkyl sulfates (e.g., sodium dodecyl sulfate and sodium hexadecyl sulfate); sodium dodecylbenzene sulfonate, sodium dioctyl sulfosuccinate; fatty acids (e.g., fatty acids derived from coconut oil); cationic surfactants, such as those of the formula N+R′R″R′″R″″ The following are examples of surfactants: water-soluble quaternary ammonium salts of Y-, wherein R- is an optionally hydroxylated hydrocarbon group and Y- is an anion of a strong acid, such as a halide, sulfate, or sulfonate anion; hexadecyltrimethylammonium bromide, which is a cationic surfactant of the formula N+R′R″R′″, wherein R- is an optionally hydroxylated hydrocarbon group (e.g., octadecylamine hydrochloride); nonionic surfactants, such as optionally polyoxyethyleneized sorbitan esters (e.g., polysorbate 80), polyoxyethyleneized alkyl ethers; polyoxypropylized fatty alcohols, such as polyoxypropylene-styrene ethers; polyethylene glycol stearate, polyoxyethyleneized derivatives of castor oil, polyglycerol esters, polyoxyethyleneized fatty alcohols, polyoxyethyleneized fatty acids, copolymers of ethylene oxide and propylene oxide; amphoteric surfactants, such as substituted dodecyl compounds of betaine; and mixtures of at least two of these agents.

[0339] The pharmaceutical compositions disclosed herein may be intended for rectal administration, for example in the form of suppositories, which will dissolve and release the drug in the rectum. Compositions for rectal administration may contain an oily matrix as a suitable non-irritating excipient. Such a matrix may include, but is not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0340] The pharmaceutical compositions of this disclosure can comprise a variety of materials that alter the physical form of solid or liquid dosage units. For example, the composition can include a material that forms a coating around the active ingredient. The material forming the coating is typically inert and can be selected from, for example, sugars, shellac, and other enteric coating agents. Alternatively, the active ingredient can be encapsulated in a gelatin capsule.

[0341] Administration

[0342] The dosage of compounds of Formula I as described herein will vary depending on the specific compound, the subject and their physical condition, the nature and severity of the disease, and the chosen route of administration. Therefore, the amount of compound or composition administered to an animal requiring treatment can vary widely. The amount of compound administered can depend on factors such as the efficacy of the compound, the type of animal being treated, the animal's weight, the animal's age, the desired effect, and the nature and severity of the disease. It should be understood that the dosage may vary depending on each subject's requirements and the severity of the disorder or disease being treated. Those skilled in the art will be able to determine the specific effective dosage. Generally, a therapeutically effective dosage is the amount of compound or composition sufficient to provide a beneficial effect or otherwise reduce harmful non-beneficial events to the subject to whom the compound or composition has been administered. A therapeutically effective dosage can be a dosage that produces one or more desired or ideal (e.g., beneficial) effects, which are the purpose of its administration, and this administration occurs once or multiple times within a given time period. Additionally, it should be understood that an initial higher dose (i.e., one or more loading doses) may be administered to rapidly achieve the desired plasma concentration. On the other hand, the initial dose may be less than the optimal dose, and the daily dose may be gradually increased during treatment (i.e., dose adjustment) depending on the specific circumstances.

[0343] The frequency and duration of administration of the compounds or compositions provided herein may vary. Administration may be intermittent in time and may be administered daily, weekly, bi-weekly, monthly, bi-monthly, quarterly, or even for longer periods. In some embodiments, administration is performed daily for durations of at least about one week, two weeks, three weeks, one month, or two months, up to six months, one year, or more, including the subject's lifetime.

[0344] In some implementations, the required dose may be administered as a single dose or as multiple doses administered at appropriate intervals, such as two, three, four, or more sub-dose per day. The sub-dose itself may be further subdivided, for example, into multiple discrete, loosely spaced administrations, such as multiple oral dosage forms.

[0345] Those skilled in the art will recognize that the optimal amount and interval of an individual dose of a compound as described herein or a composition containing a compound as described herein will be determined by the nature and extent of the condition being treated, the form, route, and site of administration, and the age and condition of the specific subject being treated, and the physician will ultimately determine the appropriate dose, frequency, and duration of treatment. The selected dose may be repeated as needed. If side effects occur, the amount and / or frequency of the dose may be modified or reduced according to normal clinical practice. Those skilled in the art will be able to develop specific administration protocols for specific situations.

[0346] Methods of treating atopic dermatitis and / or pruritus in dogs

[0347] This document provides a method for treating canine atopic dermatitis and / or pruritus. The method generally comprises administering an effective amount of a compound of this disclosure, its salts or solvates, or a composition comprising said compound or its salts or solvates. Without wishing to be bound by any particular theory, the disclosed compounds are believed to provide efficacy and tolerability in inhibiting Janus kinase 1 (JAK-1; for example, canine JAK-1 (cJAK-1)), which, together with their long half-life, allows for once-daily dosing.

[0348] The pathogenesis of canine atopic dermatitis is complex. Transdermal sensitization to environmental allergens (e.g., dust mites, pollen, mold) and / or food allergens induces skin infiltration of various inflammatory cells, activation of resident cells, and local production of inflammatory / pruritic mediators. Multiple factors can exacerbate canine atopic dermatitis, including ectoparasites (especially fleas), environmental factors, bacterial and yeast colonization / infection of the skin, and epidermal barrier dysfunction.

[0349] The JAK / STAT signaling pathway has been identified as being involved in mediating many aberrant immune responses, such as allergies, asthma, and autoimmune diseases (e.g., rheumatoid arthritis). JAK1 inhibition blocks the signaling of many important pro-inflammatory cytokines, including interleukin (IL)-2, IL-6, IL-7, and IL-15, which are known to contribute to inflammatory disorders such as atopic dermatitis. Therefore, inhibition of JAK signaling, particularly JAK-1 signaling, has been proposed for the treatment of immune-mediated disorders such as atopic dermatitis. In addition to the anti-inflammatory activity of JAK-1 inhibitors, type 2 cytokines IL-4 and IL-13 have been reported to directly stimulate sensory neurons, and chronic pruritus depends on neuronal IL-4Rα and JAK1 signaling (see, for example, U.S. Patent Application Publication No. 2021 / 0338812). Furthermore, as mentioned above, olatinib (APOQUEL) ®JAK inhibitors have been approved by the FDA for the treatment of canine atopic dermatitis, which further supports the concept of using JAK inhibitors to treat pruritus / immune / allergic disorders in companion animals. (Gonzales et al., J. Vet. Pharmacol Ther. 2014 Aug; 37(4):317-24).

[0350] Olapinib is most effective in inhibiting JAK-1 (IC50). 50 = 10 nM), and also in IC 50 Olapinib, with a range of 36 to 249 nM, inhibits the function of JAK1-dependent cytokines involved in allergy and inflammation (IL-2, IL-4, IL-6, and IL-13) and pruritus (IL-31) (Gonzales et al., 2014). Olapinib has been described as a targeted therapy that selectively inhibits JAK1-dependent cytokines involved in allergy, inflammation, and pruritus, suggesting that these are the mechanisms by which olalapinib controls clinical symptoms associated with allergic dermatitis in dogs.

[0351] It has been found that olatinib exhibits relatively low selectivity for cJAK-2, cJAK-3, and cJAK-4 compared to cJAK-1. This lack of selectivity is believed to be the cause of dose-limiting vomiting observed in dogs and may be related to cJAK-2. As described above, there is a desire in the art to provide a cJAK inhibitor that is effective in treating canine atopic dermatitis and related symptoms while allowing for once-daily (QD) dosing.

[0352] Therefore, this document discloses compounds that exhibit selectivity for JAK-1 inhibition that is approximately 10 times greater than that for JAK-2 inhibition (in dogs, humans, or both), and methods for treating canine atopic dermatitis, pruritus, or both using such compounds. The treatment typically involves administering a therapeutically effective amount of a compound of formula I, its salt, or a solvate, or a composition comprising said compound, its salt, or a solvate.

[0353] Typically, the methods include oral, parenteral, topical, rectal, or transmucosal administration of the compounds or pharmaceutical compositions thereof disclosed herein. Parenteral administration includes indirect injection to produce a systemic effect or direct injection into the affected area. Topical administration includes treatment of easily accessible skin or organs, such as the ear, by local application. It also includes transdermal delivery to produce a systemic effect. Rectal administration includes suppository forms. In some embodiments, the route of administration is oral or parenteral. In a specific embodiment, the route of administration is oral.

[0354] As described above, the frequency of administration can vary depending on the specific compound, the subject, the nature and severity of the dermatitis, etc. In some embodiments, the compound or composition is administered orally daily. Also as described above, in some embodiments, the compounds of this disclosure have pharmacological and pharmacokinetic properties suitable for once-daily administration. Therefore, in some embodiments, administration is once-daily.

[0355] Combination therapy

[0356] One or more compounds disclosed herein, or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, may be administered simultaneously, before, or after one or more other therapeutic agents. Such combination therapy may include administration of a single pharmaceutical dosage form containing one or more compounds of this disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and one or more additional active agents, or administration of one or more compounds of this disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, and each active agent in its own separate pharmaceutical dosage form. For example, one or more compounds of this disclosure or pharmaceutically acceptable salts, solvates, stereoisomers, or tautomers thereof, along with other active agents, may be administered to a subject in a single oral dose composition (e.g., tablets or capsules), or each agent may be administered in a separate oral dose formulation. When using individual dosage forms, one or more compounds of this disclosure or their pharmaceutically acceptable salts, solvates, stereoisomers or tautomers, and one or more additional active agents may be administered substantially simultaneously (e.g., concurrently) or separately at staggered times (e.g., sequentially); combination therapy should be understood to include all of these regimens.

[0357] Suitable therapeutic agents for combination therapy with the disclosed compounds include agents that modulate the mammalian immune system, antihistamines, and anti-inflammatory agents. These agents may include, but are not limited to, cyclosporine A (e.g., Sandimmune). ® or Neoral ® Rapamycin, FK-506 (tacrolimus), leflunomide, deoxyguanidine, mycophenolate (e.g., Cellcept) ® ), azathioprine (e.g., Imuram) ® ), daclizumab (e.g., Zenapax) ® OKT3 (e.g., Orthocolone) ®(e.g., AtGam, aspirin, acetaminophen, ibuprofen, naproxen, piroxicam, and anti-inflammatory steroids (e.g., prednisolone or dexamethasone). These agents may be administered as part of the same or different dosage forms, via the same or different routes of administration, and according to the same or different administration regimens, in accordance with standard pharmaceutical practice known to those skilled in the art.

[0358] Although the technology described herein has been illustrated with reference to specific embodiments, it should be understood that these embodiments are merely illustrative of the principles and applications of the technology. It will be apparent to those skilled in the art that various modifications and variations can be made to the methods and apparatus of this technology without departing from the spirit and scope of the technology. Therefore, this technology is intended to include modifications and variations within the scope of the appended claims and their equivalents. Thus, this disclosure is not limited except by the appended claims.

[0359] Throughout this specification, references to "an embodiment" or "a particular embodiment" mean that a specific feature, structure, or characteristic described in connection with said embodiment is included in at least one embodiment of this disclosure. Therefore, the phrases "in an embodiment" or "in a particular embodiment" appearing throughout the specification do not necessarily all refer to the same embodiment. Furthermore, a particular feature, structure, or characteristic may be combined in any suitable manner in one or more embodiments. Any scope referenced herein includes endpoints.

[0360] Although preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, modifications, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in carrying out the invention. The following claims are intended to define the scope of the invention, and methods and structures within the scope of these claims and their equivalents are intended to be covered therewith.

[0361] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference to the extent that each individual publication, patent or patent application is expressly and individually indicated as incorporated by reference.

[0362] Various aspects of this technology are further illustrated with reference to the following embodiments. Before describing several exemplary embodiments of this technology, it should be understood that this technology is not limited to the details of the construction or process steps set forth in the following description. This technology can have other embodiments and can be implemented or carried out in various ways. The following embodiments are intended to illustrate certain aspects of this technology and should not be construed as limiting the technology. It should be understood that those skilled in the art can prepare these compounds by similar methods or in combination with other methods known to them. It should also be understood that those skilled in the art can prepare other compounds within the scope of this disclosure in a manner similar to that described below, by using appropriate starting materials and modifying the synthetic route as needed. Typically, starting materials and reagents are available from commercial suppliers, synthesized from sources known to those skilled in the art, or prepared as described herein.

[0363] The names of the reagents / reactants given should be as indicated on the commercial vials or according to IUPAC conventions, ChemDraw 19.0 (CAMBRIDGESOFT). ® (PerkinElmer) generated. Compounds designated as salts (e.g., hydrochlorides, acetates, sulfates) may contain more than one molar equivalent of acid.

[0364] Example

[0365] Various aspects of the present invention will be more fully described through the following embodiments, which are intended to illustrate certain aspects of the invention and should not be construed as limiting the invention.

[0366] Example 1. 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N,N-dimethylacetamide N - (4- N,N - dimethyl Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(tert- butylsulfamoyl)phenyl)acetamide (Compound 1).

[0367]

[0368] Step A. Synthesis of ethyl 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)

[0369] At room temperature, ethyl 2-(piperazin-1-yl)acetate (16.86 g, 98.04 mmol, 1.50 equivalent) and DIEA (25.29 g, 196.08 mmol, 3.00 equivalent) were added to a stirred solution of 4-chloro-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetate in 1-butanol (50 mL). The resulting mixture was stirred at 90 °C for 18 hours. The mixture was concentrated, diluted with water, and extracted with 1-butanol. The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with DCM / MeOH = 10 / 1. Ethyl 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetate (18.00 g, 95.0%) was produced as a creamy white color. [M+H] + = 290.

[0370] Step B. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid

[0371] At room temperature, LiOH·H₂O (3.49 g, 83.04 mmol, 3.00 equivalent) was added to a stirred solution of ethyl 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (8.00 g, 27.68 mmol, 1.00 equivalent) in EtOH / H₂O (20 mL / 5 mL). The resulting mixture was stirred at room temperature for 2 hours. The mixture was concentrated, diluted with water, adjusted to pH 2–3 with 2N HCl solution, and then extracted with EtOAc. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to give 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (6.36 g, 88.0%) as a white solid. [M+H] + = 263.

[0372] Step C. 2-Methoxy- N,N Synthesis of 4-dimethyl-4-nitrobenzenesulfonamide

[0373] At room temperature, 2-methoxy-4-nitrobenzenesulfonyl chloride (1.00 g, 3.98 mmol, 1.00 equivalent) was added to a solution of dimethylamine in THF (15 mL, 2 M). The resulting mixture was stirred at room temperature for 30 minutes. The mixture was concentrated, diluted with NaHCO3 solution, and then extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give 2-methoxy- N,N1-Dimethyl-4-nitrobenzenesulfonamide (1.00 g, 100.0%) is a yellow solid. [M+H] + = 261.

[0374] Step D. 4-Amino-2-methoxy- N,N Synthesis of dimethylbenzenesulfonamide

[0375] At room temperature and in H2, 2-methoxy- N,N 1-Dimethyl-4-nitrobenzenesulfonamide (1.00 g, 3.85 mmol, 1.00 equivalent) was added to a solution of MeOH (15 mL) with 10% Pd / C (120.00 mg, 1.13 mmol, 0.29 equivalent). The resulting mixture was stirred at 60 °C for 3 hours. The mixture was filtered to give 4-amino-2-methoxy- N,N 1-Dimethylbenzenesulfonamide (352.00 mg, 40.0%) is a yellow solid. [M+H] + = 231.

[0376] Step E. 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)- N -(4-( N,N Synthesis of (-dimethylaminosulfonyl)-3-methoxyphenyl)acetamide (compound 1)

[0377] At room temperature, HATU (196.00 mg, 0.52 mmol, 1.20 equivalent), pyridine (102.00 mg, 1.29 mmol, 3.00 equivalent), and 4-amino-2-methoxy- N,N - Dimethylbenzenesulfonamide (100.00 mg, 0.43 mmol, 1.00 equivalent). The resulting mixture was stirred at 40 °C for 18 hours. The resulting solution was purified by preparative HPLC under the following conditions: column: RP-PREP-5 Xbridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH3H2O), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 10% B to 55% B over 11 minutes; wavelength: 214 nm. The title compound was given as a white solid (52.00 mg, 25.0%). [M+H] + = 474. 1H NMR (400 MHz, CDCl3) δ 2.71-2.82 (10H, m), 3.19 (2H, s), 3.87 (3H, s), 4.04 (4H, d, J = 4.4Hz), 6.46 (1H, d, J = 3.6 Hz), 6.76-6.79 (1H, m), 7.06 (1H, d, J = 3.6 Hz), 7.77-7.80 (2H, m), 8.29 (1H, s), 9.25 (1H, s), 10.15 (1H, s).

[0378] Example 2. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(tert- butylsulfamoyl)phenyl)acetamide (Compound 2) Example 3. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(tert- butylsulfamoyl)phenyl)acetamide (Compound 3) .

[0379]

[0380] Step A. Synthesis of N-(tert-butyl)-4-nitrobenzenesulfonamide

[0381] Add 4-nitrobenzenesulfonyl chloride (500.00 mg, 2.26 mmol, 1.00 equivalent) and 2-methylpropyl-2-amine (330.00 mg, 4.52 mmol, 2.00 equivalent) to a solution of THF (10 mL) under stirring. Stir the resulting mixture at 30 °C for 1 hour. LCMS showed the reaction was complete. Wash the resulting solution with water and extract with EA (10 mL × 3). Remove the solvent to give the desired product. N-(tert-butyl)-4-nitrobenzenesulfonamide (532.00 mg, 91.00%) is produced as a yellow solid. [M+H] + =259.

[0382] Step B. Synthesis of 4-amino-N-(tert-butyl)benzenesulfonamide

[0383] To a stirred solution of N-(tert-butyl)-4-nitrobenzenesulfonamide (532.00 mg, 2.06 mmol, 1.00 equivalent) in EtOH (20 mL), 10% Pd / C (50 mg) was added. The resulting mixture was stirred overnight at 60 °C under a H2 atmosphere. LC-MS showed that the reaction was complete. The resulting solution was filtered to remove the solvent, giving the title compound (600.00 mg, 100.00%) as a yellow oil. [M+H] + =229.

[0384] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(tert-butyl)aminosulfonyl)phenyl)acetamide (compound 2)

[0385] To a stirred solution of 4-amino-N-(tert-butyl)benzenesulfonamide (100.00 mg, 0.44 mmol, 1.00 equivalent) in DMF (5 mL), 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (137.00 mg, 0.53 mmol, 1.20 equivalent), pyridine (103.90 mg, 1.32 mmol, 3.00 equivalent), and HATU (333.00 mg, 0.88 mmol, 2.00 equivalent) were added. The resulting mixture was stirred at 40 °C for 18 hours. LCMS showed the desired product. The mixture was concentrated under vacuum, and the residue was purified by preparative HPLC under the following conditions: column: XBridge C18SN.271 / 3611211207 waters, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 50% B over 11 min; wavelength: 214 nm. The title compound was given as a white solid (38.00 mg, 18%). [M+H] + = 472. 1 H NMR (400 MHz, DMSO- d 6 ): δ 1.08 (9H, s), 2.65-2.67 (4H, m), 3.24 (2H, s), 3.93-3.95 (4H, m), 6.62 (1H, s), 7.18 (1H, s),7.39 (1H, s), 7.76 (2H, d, J = 8.8 Hz), 7.83 (2H, d, J = 8.8 Hz), 8.14 (1H, s), 10.13 (1H, s), 11.69 (1H, s).

[0386] Example 4. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(azetidin- 1-ylsulfonyl)phenyl)acetamide (Compound 4) Example 5. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N,N- dimethylsulfamoyl)-3-fluorophenyl)acetamide (Compound 5)

[0387]

[0388] Step A. Synthesis of N-ethyl-4-nitrobenzenesulfonamide

[0389] Ethylamine (2.0 mol / L, 20 mL, 8.85 equivalents) was added to a stirred solution of 4-nitrobenzenesulfonyl chloride (1.00 g, 4.52 mmol, 1.00 equivalents) in THF. The resulting mixture was stirred at 30 °C for 1 hour. LC-MS showed the reaction was complete. The resulting solution was washed with water and extracted with EA (10 mL × 3). The solvent was removed to give N-ethyl-4-nitrobenzenesulfonamide (1.05 g, 100%) as a yellow solid. [M+Na] + =253.

[0390] Step B. Synthesis of N-ethyl-N-methyl-4-nitrobenzenesulfonamide

[0391] To a stirred solution of N-ethyl-4-nitrobenzenesulfonamide (500.00 mg, 2.17 mmol, 1.00 equivalent) in THF (10 mL), 60% NaH (174.00 mg, 2.00 equivalent) and methyl iodide (617.00 mg, 4.35 mmol, 2.00 equivalent) were added. The resulting mixture was stirred at 30 °C for 30 min. LCMS showed the reaction was complete. The resulting solution was washed with water and extracted with EA (10 mL × 3). The solvent was removed to give N-ethyl-N-methyl-4-nitrobenzenesulfonamide (930 mg, 100%) as a yellow solid. [M+H] + =245.

[0392] Step C. Synthesis of 4-amino-N-ethyl-N-methylbenzenesulfonamide

[0393] To a stirred solution of N-ethyl-N-methyl-4-nitrobenzenesulfonamide (930.00 mg, 3.8 mmol, 1.00 equivalent) in EtOH (20 mL), 10% Pd / C (93 mg) was added. The resulting mixture was stirred overnight at 60 °C under a H2 atmosphere. LCMS showed that the reaction was complete. The resulting solution was filtered to remove the solvent, yielding 4-amino-N-ethyl-N-methylbenzenesulfonamide (500.00 mg, 60.00%) as a yellow oil. [M+H] + =215.

[0394] Step D. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(tert-butyl)aminosulfonyl)phenyl)acetamide (compound 3)

[0395] To a stirred solution of 4-amino-N-ethyl-N-methylbenzenesulfonamide (100.00 mg, 0.47 mmol, 1.00 equivalent) in DMF (5 mL), 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (146.00 mg, 0.56 mmol, 1.20 equivalent), pyridine (111.00 mg, 1.40 mmol, 3.00 equivalent), and HATU (533.00 mg, 1.40 mmol, 3.00 equivalent) were added. The resulting mixture was stirred at 45 °C for 18 hours. LCMS showed the desired product. The residue was purified by preparative HPLC under the following conditions: column: XBridge C18 SN.271 / 3611211207 waters, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 25% B to 50% B over 11 min; wavelength: 214 nm. The title compound (48.00 mg, 22%) was given as a white solid. [M+H] + = 458. 1 H NMR (400 MHz, CDCl3): δ 1.16 (3H, t, J = 7.2 Hz), 2.76 (3H, s), 2.81-2.84 (4H, m), 3.10-3.15 (2H, m), 3.28 (2H, s), 4.10-4.12 (4H, m), 6.54 (1H,s), 7.17 (1H, s), 7.76-7.81 (4H, m), 8.40 (1H, s), 9.39 (1H, s), 10.84 (1H, s).

[0396] Example 6. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(3-chloro-4- (N,N-dimethylsulfamoyl)phenyl)acetamide (Compound 6) N,N

[0397]

[0398] Step A. Synthesis of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine

[0399] At room temperature, piperazine (1.44 g, 16.70 mmol, 1.00 equivalent) and DIEA (3.23 g, 25.05 mmol, 1.50 equivalent) were added to a stirred solution of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (2.56 g, 16.70 mmol, 1.00 equivalent) in 1-butanol (15 mL). The resulting mixture was stirred at 90 °C for 18 hours. The mixture was filtered and concentrated to give 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (3.25 g, 96.0%) as a white solid. [M+H] + =204.

[0400] Step B. Synthesis of 1-((4-nitrophenyl)sulfonyl)azacyclobutane

[0401] At room temperature, aziridine (0.63 g, 6.79 mmol, 1.50 equivalent) and DIEA (1.75 g, 13.56 mmol, 3.00 equivalent) were added to a stirred solution of 4-nitrobenzenesulfonyl chloride (1.00 g, 4.52 mmol, 1.00 equivalent) in 15 mL of THF. The resulting mixture was stirred at room temperature for 2 hours. The mixture was diluted with water and then extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give 1-((4-nitrobenzene)sulfonyl)aziridine (670 mg, 61.0%) as a yellow solid. [M+H] + =243.

[0402] Step C. Synthesis of 4-(azacyclobutane-1-ylsulfonyl)aniline

[0403] At room temperature, Fe (231.00 mg, 4.13 mmol, 10.00 equivalent) and NH4Cl (110.00 mg, 2.07 mmol, 5.00 equivalent) were added to a solution of 1-((4-nitrophenyl)sulfonyl)azacyclobutane (100.00 mg, 0.41 mmol, 1.00 equivalent) in EtOH / H2O (12 / 3 mL). The resulting mixture was stirred at 80 °C for 1 hour. The mixture was filtered and concentrated, diluted with NaHCO3 solution, and then extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give 4-(azacyclobutane-1-ylsulfonyl)aniline (60.00 mg, 68.0%) as a yellow solid. [M+H] + =213.

[0404] Step D. Synthesis of N-(4-(azacyclobutane-1-ylsulfonyl)phenyl)-2-chloroacetamide

[0405] At room temperature, pyridine (44.00 mg, 0.56 mmol, 2.00 equivalent) and DMAP (3.00 mg, 0.03 mmol, 0.10 equivalent) were added to a stirred solution of 4-(azacyclobutane-1-ylsulfonyl)aniline (60.00 mg, 0.28 mmol, 1.00 equivalent) in DCM (10 mL), followed by 2-chloroacetyl chloride (32.00 mg, 0.28 mmol, 1.00 equivalent). The resulting mixture was stirred at room temperature for 15 minutes. The mixture was diluted with H2O and extracted with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give N-(4-(azacyclobutane-1-ylsulfonyl)phenyl)-2-chloroacetamide (81.00 mg, 100%) as a yellow solid. [M+H] + =289.

[0406] Step E. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(azacyclobutane-1-ylsulfonyl)phenyl)acetamide (compound 4)

[0407] At room temperature, 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (57.00 mg, 0.28 mmol, 1.00 equivalent), TEA (141.00 mg, 1.40 mmol, 5.00 equivalent), and KI (23.00 mg, 0.14 mmol, 0.50 equivalent) were added to a solution of N-(4-(azacyclobutan-1-ylsulfonyl)phenyl)-2-chloroacetamide (81.00 mg, 0.28 mmol, 1.00 equivalent) in DMF (10 mL). The resulting mixture was stirred at 80 °C for 1 hour. The resulting solution was purified using preparative HPLC under the following conditions: column: RP-PREP-5 Xbridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 35% B to 65% B over 10 min; wavelength: 214 nm. The title compound (46.00 mg, 36.0%) was obtained as a white solid. [M+H] + = 456. 1 H NMR (400 MHz, CDCl3): δ 2.06-2.14 (2H, m), 2.84 (4H, t, J= 5.2 Hz), 3.30 (2H, s), 3.78-3.82 (4H,m), 4.10- 4.13 (4H, m), 6.55 (1H, d, J = 4.0 Hz), 7.15 (1H, d, J = 3.6 Hz), 7.88-7.83 (4H, m), 8.39 (1H, s), 9.44 (1H, s), 10.23 (1H, s).

[0408] N,N N,N

[0409]

[0410] Step A. Synthesis of 2-fluoro-N,N-dimethyl-4-nitrobenzenesulfonamide

[0411] To a solution of 2-fluoro-4-nitrobenzenesulfonyl chloride (300.00 mg, 1.25 mmol, 1.00 equivalent) and dimethylamine hydrochloride (112.75 mg, 1.37 mmol, 1.10 equivalent) in DCM (10 mL), DIEA (322.00 mg, 2.50 mmol, 2.00 equivalent) was added. The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was complete. The residue was extracted with DCM and washed with brine. The organic layer was dried over Na₂SO₄ and concentrated to give 2-fluoro-N,N-dimethyl-4-nitrobenzenesulfonamide (280 mg, 90.3%) as a creamy white solid. [M+H] + =249.

[0412] Step B. Synthesis of 4-amino-2-fluoro-N,N-dimethylbenzenesulfonamide

[0413] To a solution of 2-fluoro-N,N-dimethyl-4-nitrobenzenesulfonamide (130.00 mg, 0.52 mmol, 1.00 equivalent) in EtOH / water (10 mL / 2 mL), Fe (293.00 mg, 5.20 mmol, 10.00 equivalent) and NH4Cl (140.00 mg, 2.60 mmol, 5.00 equivalent) were added. The mixture was stirred at 80 °C for 1 hour. LCMS showed the reaction was complete. The mixture was filtered, and the filtrate was extracted with EtOAc and washed with water. The organic layer was dried over Na2SO4 and concentrated to give 4-amino-2-fluoro-N,N-dimethylbenzenesulfonamide (161.00 mg, >100%) as a gray solid. [M+H] + =219.

[0414] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N,N-dimethylaminosulfonyl)-3-fluorophenyl)acetamide (compound 5)

[0415] To a solution of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (90.00 mg, 0.34 mmol, 1.00 equivalent) in DCM (10 mL), add (COCl)₂ (131.00 mg, 1.03 mmol, 3.00 equivalent) and one drop of DMF. Stir the mixture at room temperature for 2 hours and concentrate to obtain the residue. To a solution of 4-amino-2-fluoro-N,N-dimethylbenzenesulfonamide (90.00 mg, 0.41 mmol, 1.20 equivalent) and DIEA (88.70 mg, 0.69 mmol, 2.00 equivalent) in DCM (10 mL), add the solution of the above residue in DCM. Stir the mixture at 80 °C for 3 hours. LCMS showed the reaction was complete. Concentrate the mixture. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 25% B to 40% B over 12 min; wavelength: 214 nm. The title compound (6.00 mg, 3.1%) was obtained as a creamy white solid. [M+H] + =462. 1 H NMR (400 MHz, DMSO- d 6 ): δ 2.64-2.69 (10H, m), 3.27 (2H, s), 3.94 (4H, t, J = 4.8 Hz), 6.62 (1H, d, J = 3.2 Hz), 7.19 (1H, d, J = 3.6 Hz),7.62-7.65 (1H, m), 7.73 (1 H, t, J = 8.8 Hz), 7.88-7.92 (1 H, m), 8.15 (1 H, s), 10.42 (1 H, s), 11.70 (1 H, s).

[0416] N,N Example 7. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- cyclopropylsulfamoyl)phenyl)acetamide (Compound 7)

[0417]

[0418] Step A. Synthesis of 2-chloro-N,N-dimethyl-4-nitrobenzenesulfonamide

[0419] To a solution of 2-chloro-4-nitrobenzenesulfonyl chloride (512.00 mg, 2.00 mmol, 1.00 equivalent) and dimethylamine hydrochloride (200.00 mg, 2.40 mmol, 1.20 equivalent) in DCM (15 mL), DIEA (516.00 mg, 4.00 mmol, 2.00 equivalent) was added. The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was complete. The residue was extracted with DCM and washed with brine. The organic layer was dried over Na2SO4 and concentrated to give 2-chloro- Example 8. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- methylsulfamoyl)phenyl)acetamide (Compound 8) Dimethyl-4-nitrobenzenesulfonamide (518.00 mg, 98.1%) is a creamy white solid. [M+H] + =265.

[0420] Step B. 4-Amino-2-chloro- Example 9. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(3-fluoro-4- (N-methylsulfamoyl)phenyl)acetamide (Compound 9) Synthesis of dimethylbenzenesulfonamide

[0421] Fe (1.10 g, 19.60 mmol, 10.00 equivalent) and NH₄Cl (520.00 mg, 9.80 mmol, 5.00 equivalent) were added to a solution of 2-chloro-N,N-dimethyl-4-nitrobenzenesulfonamide (518.00 mg, 1.96 mmol, 1.00 equivalent) in EtOH / water (10 mL / 3 mL). The mixture was stirred at 80 °C for 1 hour. LCMS showed that the reaction was complete. The mixture was filtered, and the filtrate was extracted with DCM and washed with water. The organic layer was dried over Na₂SO₄ and concentrated to give 4-amino-2-chloro-N,N-dimethylbenzenesulfonamide (440.00 mg, 96.1%) as a gray solid. [M+H] + =235.

[0422] Step C. 2-Chloro-N-(3-Chloro-4-( Example 10. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(2- methoxyethyl)-N-methylsulfamoyl)phenyl)acetamide (Compound 10) Synthesis of 1,2-dimethylaminosulfonyl)phenyl)acetamide

[0423] To 4-amino-2-chloro- Example 11. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- butylsulfamoyl)phenyl)acetamide (Compound 11)A solution of dimethylbenzenesulfonamide (200.00 mg, 0.85 mmol, 1.00 equivalent), pyridine (135.00 mg, 1.71 mmol, 2.00 equivalent), and DMAP (10.00 mg, 0.08 mmol, 0.10 equivalent) in DCM (10 mL) was mixed with 2-chloroacetyl chloride (97.00 mg, 0.85 mmol, 1.00 equivalent). The mixture was stirred at room temperature for 1 hour. LCMS showed that the reaction was complete. The residue was extracted with DCM and washed with brine. The organic layer was dried over Na2SO4 and concentrated to give 2-chloro-N-(3-chloro-4-(N,N-dimethylaminosulfonyl)phenyl)acetamide (304 mg, >100%) as a brown solid. [M+H] + =311.

[0424] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(3-chloro-4-(N,N-dimethylaminosulfonyl)phenyl)acetamide (compound 6)

[0425] To a solution of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (HCl salt; 80.00 mg, 0.39 mmol, 1.00 equivalent) and 2-chloro-N-(3-chloro-4-(N,N-dimethylaminosulfonyl)phenyl)acetamide (121 mg, 0.39 mmol, 1.00 equivalent) in EtOH (10 mL), TEA (196.00 mg, 1.96 mmol, 5.00 equivalent) and KI (33.00 mg, 0.20 mmol, 0.50 equivalent) were added. The mixture was stirred at 80 °C for 2 hours. LCMS showed the reaction was complete. The mixture was then concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH3·H2O), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 10% B to 55% B over 10 min; wavelength: 214 nm. The title compound (43 mg, 23%) was given as a white solid. [M+H] + =478. 1 HNMR (400 MHz, DMSO- d 6 ): δ 2.65 (4H, t, J = 4.4 Hz), 2.77 (6H, s), 3.26 (2H,s), 3.94 (4H, t, J= 4.4 Hz), 6.61 (1H, d, J = 3.2 Hz), 7.18 (1H, d, J = 3.2Hz), 7.80-7.82 (1H, m), 7.90 (1H, d, J = 8.8 Hz), 8.09 (1H, d, J = 2.4 Hz), 8.15 (1H, s), 10.34 (1H, s), 11.70 (1H, s).

[0426] Example 14. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- propylsulfamoyl)phenyl)acetamide (Compound 12) ​

[0427]

[0428] Step A. Synthesis of N-cyclopropyl-4-nitrobenzenesulfonamide

[0429] To a solution of cyclopropylamine (130 mg, 2.20 mmol, 1.00 equivalent) in DCM / Et3N (10 ml / ml), 4-nitrobenzenesulfonyl chloride (500 mg, 2.20 mmol, 1.00 equivalent) was added. The resulting mixture was stirred at room temperature for 1 hour and monitored by TLC. The mixture was extracted with ethyl acetate and water, and the combined organic layers were evaporated under reduced pressure to give crude N-cyclopropyl-4-nitrobenzenesulfonamide (530.00 mg, 96.36%) as a yellow oil. [M+H] + = N / A

[0430] Step B. Synthesis of 4-amino-N-cyclopropylbenzenesulfonamide

[0431] Under H2 balloon conditions, N-cyclopropyl-4-nitrobenzenesulfonamide (530 mg, 2.19 mmol, 1.00 equivalent) in MeOH (30 mL) was added to a flask. The reaction mixture was stirred at room temperature for 16 hours, monitored by LC-MS. After the starting material was consumed, the mixture was filtered and evaporated under reduced pressure to give the crude title product (500.00 mg, >100%) as a gray solid. [M+H] + = 213

[0432] Step C. Synthesis of 2-chloro-N-(4-(N-cyclopropylaminosulfonyl)phenyl)acetamide

[0433] 2-Chloroacetyl chloride (30 mg, 0.23 mmol, 1.00 equivalent) was added to a solution (50 mg, 0.23 mmol, 1.00 equivalent) in DCM / Et3N (5 ml / 0.2 ml). The resulting mixture was stirred at room temperature for 1 hour and monitored by TLC. The mixture was extracted with ethyl acetate and water, and the combined organic layers were evaporated under reduced pressure to give crude 2-chloro-N-(4-(N-cyclopropylaminosulfonyl)phenyl)acetamide (80 mg, >100.00%) as a yellow oil. [M+H] + = N / A

[0434] Step D. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-cyclopropylaminosulfonyl)phenyl)acetamide (compound 7)

[0435] To a solution of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (80.00 mg, 0.27 mmol, 1.10 equivalent) and 2-chloro-N-(4-(N-cyclopropylaminosulfonyl)phenyl)acetamide (50.00 mg, 0.27 mmol, 1.00 equivalent) in DMF (8 mL), K₂CO₃ (100.00 mg, 0.81 mmol, 3.00 equivalent) and KI (20.00 mg, 0.13 mmol, 0.50 equivalent) were added. The mixture was stirred at 80 °C for 1 hour. LCMS showed the reaction was complete. The mixture was then concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; gradient: 15% B to 60% B over 10 min; flow rate: 15 mL / min; wavelength: 214 nm. The title compound (20 mg, 15%) was given as a white solid. [M+H] = 456. 1 H NMR (400 MHz, DMSO- d 6 ): δ 0.34-0.37 (2H, m), 0.44-0.49 (2H, m), 2.07-2.11 (1H, m), 2.65-2.67 (4H, m), 3.26 (2H, s), 3.93-3.95 (4H, m), 6.61-6.62 (1H, m), 7.18-7.19(1H, m), 7.74-7.76 (3H, m), 7.88 (2H, d, J= 8.8 Hz), 8.15 (1H, s), 10.18(1H, s), δ 11.69 (1H, s).

[0436] ​ ​

[0437]

[0438] Step A. Synthesis of N-methyl-4-nitrobenzenesulfonamide

[0439] To a solution of methylamine (500 mg, 1.30 mmol, 1.00 equivalent) in DCM / Et3N (10 ml / 1 ml), 4-nitrobenzenesulfonyl chloride (300 mg, 1.30 mmol, 1.00 equivalent) was added. The resulting mixture was stirred at room temperature for 1 hour and monitored by TLC. The mixture was extracted with ethyl acetate and water, and the combined organic layers were evaporated under reduced pressure to give crude N-cyclopropyl-4-nitrobenzenesulfonamide (280.00 mg, 96.55%) as a yellow oil. [M+H] + = N / A.

[0440] Step B. Synthesis of 4-amino-N-methylbenzenesulfonamide

[0441] Under H2 balloon conditions, N-cyclopropyl-4-nitrobenzenesulfonamide (280 mg, 1.30 mmol, 1.00 equivalent) and 10% Pd / C (60.00 mg) in MeOH (30 mL) were added to a flask. The reaction mixture was stirred at room temperature for 16 hours, monitored by LC-MS. After the starting material was consumed, the mixture was filtered and evaporated under reduced pressure to give crude 4-amino-N-methylbenzenesulfonamide (240 mg, >100.00%) as a yellow solid. [M+H] + = 187.

[0442] Step C. Synthesis of 2-chloro-N-(4-(N-methylaminosulfonyl)phenyl)acetamide

[0443] To a solution of 4-amino-N-methylbenzenesulfonamide (100.00 mg, 0.53 mmol, 1.00 equivalent) in DCM / Et3N (8 ml / 0.2 ml), 2-chloroacetyl chloride (60.00 mg, 0.53 mmol, 1.00 equivalent) was added. The resulting mixture was stirred at room temperature for 1 hour and monitored by TLC. The mixture was extracted with ethyl acetate and water, and the combined organic layers were evaporated under reduced pressure to give crude 2-chloro-N-(4-(N-methylaminosulfonyl)phenyl)acetamide (60 mg, 42.80%) as a yellow oil. [M+H] + = 263.

[0444] Step D. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-methylaminosulfonyl)phenyl)acetamide (compound 8)

[0445] To a solution of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (47 mg, 0.22 mmol, 1.00 equivalent) and 2-chloro-N-(4-(N-cyclopropylaminosulfonyl)phenyl)acetamide (60 mg, 0.22 mmol, 1.00 equivalent) in DMF (8 mL), K₂CO₃ (100 mg, 0.66 mol, 3.00 equivalent) and KI (20 mg, 0.13 mmol, 0.5 equivalent) were added. The mixture was stirred at 80 °C for 1 hour. LCMS showed the reaction was complete. The mixture was then concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; gradient: 20% B to 65% B over 10 min; flow rate: 15 mL / min; wavelength: 214 nm. The title compound (20 mg, 15%) was given as a white solid. [M+H] = 430. 1 H NMR (400 MHz, DMSO- d 6 ): 2.39(3H, d, J = 5.2 Hz), 2.64-2.66 (4H, m), 3.25 (2H, s), 3.92-3.95 (4H, s), 6.61-6.62 (1H, m), 7.18-7.19 (1H, m), 7.31 (1H, d, J= 5.2 Hz), 7.71-7.73(2H, m), 7.86-7.88 (2H, m), 8.17 (1H, s), 10.18 (1H, s), δ 11.70 (1H, s).

[0446] ​ ​

[0447]

[0448] Step A. Synthesis of 2-fluoro-N-methyl-4-nitrobenzenesulfonamide

[0449] To a solution of 2-fluoro-4-nitrobenzenesulfonyl chloride (400.00 mg, 1.67 mmol, 1.00 equivalent) and methylamine hydrochloride (134.00 mg, 2.0 mmol, 1.20 equivalent) in DCM (10 mL), DIEA (431.00 mg, 3.34 mmol, 2.00 equivalent) was added. The mixture was stirred at room temperature for 2 hours. LCMS showed the reaction was complete. The residue was extracted with DCM and washed with brine. The organic layer was dried over Na2SO4 and concentrated. The residue was purified by column elution with PE / EA = 4 / 1 to give 2-fluoro-N-methyl-4-nitrobenzenesulfonamide (103 mg, 26.4%) as a yellow solid. [M+H] + =235.

[0450] Step B. Synthesis of 4-amino-2-fluoro-N-methylbenzenesulfonamide

[0451] To a solution of 2-fluoro-N-methyl-4-nitrobenzenesulfonamide (101.00 mg, 0.43 mmol, 1.00 equivalent) in EtOH / water (5 mL / 1 mL), Fe powder (241 mg, 4.30 mmol, 10.00 equivalent) and NH4Cl (115.00 mg, 2.14 mmol, 5.00 equivalent) were added. The mixture was stirred at 80 °C for 1 hour. LCMS showed that the reaction was complete. The mixture was filtered, and the filtrate was extracted with DCM and washed with water. The organic layer was dried over Na2SO4 and concentrated to give 4-amino-2-fluoro-N-methylbenzenesulfonamide (81 mg, 93.1%) as a yellow solid. [M+H] + =205.

[0452] Step C. Synthesis of 2-chloro-N-(3-fluoro-4-(N-methylaminosulfonyl)phenyl)acetamide

[0453] To a solution of 4-amino-2-fluoro-N-methylbenzenesulfonamide (80 mg, 0.39 mmol, 1.00 equivalent), pyridine (53.70 mg, 0.68 mmol, 2.00 equivalent), and DMAP (4.76 mg, 0.04 mmol, 0.10 equivalent) in DCM (10 mL), 2-chloroacetyl chloride (44 mg, 0.39 mmol, 1.00 equivalent) was added. The mixture was stirred at room temperature for 1 hour. LC-MS showed the reaction was complete. The residue was extracted with DCM and washed with brine. The organic layer was dried over Na2SO4 and concentrated to give 2-chloro-N-(3-fluoro-4-(N-methylaminosulfonyl)phenyl)acetamide (135 mg, >100%) as a yellow solid. [M+H] + =281.

[0454] Step D. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(3-fluoro-4-(N-methylaminosulfonyl)phenyl)acetamide (compound 9)

[0455] To a solution of 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (HCl salt) (127.00 mg, 0.30 mmol, 1.10 equivalent) and 2-chloro-N-(3-fluoro-4-(N-methylaminosulfonyl)phenyl)acetamide (135.00 mg, 0.48 mmol, 1.00 equivalent) in DMF (10 mL), DIEA (186.00 mg, 1.44 mmol, 3.00 equivalent) and KI (40.00 mg, 0.24 mmol, 0.50 equivalent) were added. The mixture was stirred at 80 °C for 3 hours. LCMS showed the reaction was complete. The mixture was then concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 10% B to 65% B over 10 min; wavelength: 214 nm. The title compound (62.00 mg, 29%) was given as a white solid. [M+H] + =448. 1 H NMR (400 MHz, DMSO- d 6 ): δ 2.46 (3H, d, J = 4.4 Hz), 2.65 (4H, t, J=4.8 Hz), 3.27 (2H, s), 3.94 (4H, t, J = 4.4 Hz), 6.61-6.62 (1H, m), 7.18-7.20(1H, m), 7.57-7.60 (2H, m), 7.69-7.75 (1H, m), 7.84-7.88 (1H, m), 8.15 (1H,s), 10.35 (1H, s), 11.71 (1H, s).

[0456] ​ ​

[0457]

[0458] Step A. Synthesis of N-(2-methoxyethyl)-N-methyl-4-nitrobenzenesulfonamide

[0459] At 0 °C, TEA (1.83 g, 18.10 mmol, 2.00 equivalent) and 2-methoxy-N-methylethyl-1-amine (1.21 g, 13.58 mmol, 1.50 equivalent) were added to a solution of 4-nitrobenzenesulfonyl chloride (2.00 g, 9.05 mmol, 1.00 equivalent) in DCM (30 mL). The resulting mixture was stirred at 0 °C for 1 hour. The residue was purified by silica gel column chromatography (PE / EtOAc = 0–30%) to give N-(2-methoxyethyl)-N-methyl-4-nitrobenzenesulfonamide (2.40 g, 96.8%) as a yellow solid.

[0460] Step B. Synthesis of 4-amino-N-(2-methoxyethyl)-N-methylbenzenesulfonamide

[0461] At room temperature, Pd / C (100 mg) was added to a solution of N-(2-methoxyethyl)-N-methyl-4-nitrobenzenesulfonamide (500 mg, 1.82 mmol, 1.00 equivalent) in EtOH (15 mL). The resulting mixture was stirred at room temperature in H2 (1 atm) for 1 hour. The mixture was filtered and dried to give 4-amino-N-(2-methoxyethyl)-N-methylbenzenesulfonamide (430 mg, 96.6%) as a yellow solid, which was used directly in the next step without further purification. [M+H] + = 245.

[0462] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-(2-methoxyethyl)-N-methylaminosulfonyl)phenyl)acetamide (compound 10)

[0463] At room temperature, pyridine (120.10 mg, 1.52 mmol, 4.00 equivalent) and 4-amino-N-(2-methoxyethyl)-N-methylbenzenesulfonamide (92.72 mg, 0.38 mmol, 1.00 equivalent) were added to a stirred solution of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (100 mg, 0.38 mmol, 1.00 equivalent) and HATU (216.60 mg, 0.57 mmol, 1.50 equivalent) in DMF (15 mL). The resulting mixture was stirred at 40 °C for 1 hour. The DMF was removed, the mixture was extracted with EA, dried over Na2SO4, and filtered. The resulting solution was purified by preparative HPLC under the following conditions: column: RP-PREP-1 XBridge C18 column, 90 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 40% B over 10 min; wavelength: 214 nm. The title compound (35 mg, 18.8%) was obtained as a white solid. [M+H] + = 488, 1 H NMR (400 MHz, DMSO- d 6 ): δ 2.50-2.69 (7H,m), 3.10 (2H, t, J = 5.6 Hz), 3.23-3.25 (5H, m), 3.44 (2H, t, J = 5.6 Hz), 3.94 (4H, t, J = 4.4 Hz), 6.62 (1H, d, J = 1.6 Hz), 7.18 (1H, t, J = 2.8 Hz), 7.73 (2H, d, J = 8.8 Hz), 7.90 (2H, d, J = 8.4 Hz), 8.14 (1H, s), 10.22 (1H, s), 11.69 (1H, s).

[0464] ​​

[0465]

[0466] Step A. Synthesis of N-butyl-4-nitrobenzenesulfonamide

[0467] At 0 °C, TEA (1.83 g, 18.10 mmol, 2.00 equivalent) and butyl-1-amine (990.98 mg, 13.58 mmol, 1.50 equivalent) were added to a solution of 4-nitrobenzenesulfonyl chloride (2.00 g, 9.05 mmol, 1.00 equivalent) in DCM (30 mL). The resulting mixture was stirred at 0 °C for 1 hour. The residue was purified by silica gel column chromatography (PE / EtOAc = 0–30%) to give N-butyl-4-nitrobenzenesulfonamide (900 mg, 38.6%) as a yellow solid.

[0468] Step B. Synthesis of 4-amino-N-butylbenzenesulfonamide

[0469] At room temperature, Fe powder (977 mg, 17.45 mmol, 5.00 equivalent) and NH4Cl (1.88 g, 34.90 mmol, 10.00 equivalent) were added to a solution of N-butyl-4-nitrobenzenesulfonamide (900 mg, 3.49 mmol, 1.00 equivalent) in EtOH (30.00 mL) and H2O (10.00 mL). The resulting mixture was stirred at 80 °C for 30 min. The mixture was cooled to room temperature, filtered, the solvent was removed under vacuum, and then extracted with EA. After drying with Na2SO4, the mixture was filtered and dried to give 4-amino-N-butylbenzenesulfonamide (780.00 mg, 97.7%) as a yellow solid, which could be used directly in the next step without further purification. [M+H] + = 229.

[0470] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-butylaminosulfonyl)phenyl)acetamide (compound 11)

[0471] At room temperature, pyridine (120.10 mg, 1.52 mmol, 4.00 equivalent) and 4-amino-N-butylbenzenesulfonamide (86.60 mg, 0.38 mmol, 1.00 equivalent) were added to a stirred solution of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (100 mg, 0.38 mmol, 1.50 equivalent) and HATU (216.60 mg, 0.57 mmol, 1.50 equivalent) in DMF (15 mL). The resulting mixture was stirred at 40 °C for 1 hour. The DMF was removed, the mixture was extracted with EA, dried over Na2SO4, and filtered. The resulting solution was purified by preparative HPLC under the following conditions: column: RP-PREP-1XBridge C18 column, 90 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 30% B to 42% B over 12 min; wavelength: 214 nm; the title compound (35 mg, 19.4%) was obtained as a white solid. [M+H] + = 472. 1 H NMR (400 MHz, DMSO- d 6 ): δ 0.79 (3H, t, J = 7.2 Hz),1.20-1.25 (2H, m), 1.30-1.35 (2H, m), 2.65-2.72 (6H, m), 3.25 (2H, s), 3.93(4H, d, J = 5.4 Hz), 6.62 (1H, t, J = 1.6 Hz), 7.18 (1H, t, J = 2.4 Hz), 7.42 (1H, t, J = 5.6 Hz), 7.73 (2H, d, J = 8.8 Hz), 7.86 (2H, d, J = 8.8 Hz), 8.17 (1H, s), 10.16 (1H, s), 11.69 (1H, s).

[0472] ​ ​

[0473]

[0474] Step A. Synthesis of 4-nitro-N-propylbenzenesulfonamide

[0475] At 0 °C, TEA (3.03 g, 30.00 mmol, 3.00 equivalent) and propan-1-amine (0.88 g, 15.00 mmol, 1.50 equivalent) were added to a solution of 4-nitrobenzenesulfonyl chloride (2.20 g, 10.00 mmol, 1.00 equivalent) in DCM (50 mL). The resulting mixture was stirred at 0 °C for 2 hours. The residue was purified by silica gel column chromatography (PE / EtOAc = 0–30%) to give 4-nitro-N-propylbenzenesulfonamide (2.20 g, 90.5%) as a yellow solid.

[0476] Step B. Synthesis of 4-amino-N-propylbenzenesulfonamide

[0477] At room temperature, Fe (2.52 g, 45.00 mmol, 5.00 equivalent) and NH4Cl (4.78 g, 90.00 mmol, 10.00 equivalent) were added to a solution of 4-nitro-N-propylbenzenesulfonamide (2.20 g, 9.00 mmol, 1.00 equivalent) in EtOH (30.00 mL) and H2O (10.00 mL). The resulting mixture was stirred at 80 °C for 1 hour. The mixture was cooled to room temperature, filtered, the solvent was removed under vacuum, and then extracted with EA. After drying with Na2SO4, the mixture was filtered and dried to give 4-amino-N-propylbenzenesulfonamide (1.70 g, 89.5%) as a white solid, which could be used directly in the next step without further purification. [M+H] + = 215.

[0478] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-propylaminosulfonyl)phenyl)acetamide (compound 12)

[0479] At room temperature, pyridine (186.45 mg, 2.36 mmol, 4.00 equivalent) and 4-amino-N-propylbenzenesulfonamide (126.26 mg, 0.59 mmol, 1.00 equivalent) were added to a stirred solution of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (154 mg, 0.59 mmol, 1.00 equivalent) and HATU (672.64 mg, 1.77 mmol, 3.00 equivalent) in DMF (15 mL). The resulting mixture was stirred at 40 °C for 18 hours. The DMF was removed, the mixture was extracted with EA, dried over Na2SO4, and filtered. The resulting solution was purified by preparative HPLC under the following conditions: column: RP-PREP-1XBridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 25% B to 45% B over 11 min; wavelength: 214 nm. The title compound (38.00 mg, 14.1%) was obtained as a white solid. [M+H] + = 458, 1 H NMR (400 MHz, DMSO- d 6 ): δ 0.79 (3H, t, J =7.6 Hz), 1.33-1.39 (2H, m), 2.65-2.69 (6H, m), 3.25 (2H, s), 3.93-3.95 (4H,m), 6.62 (1H, t, J = 1.6 Hz), 7.18 (1H, t, J = 2.4 Hz), 7.44 (1H, t, J =6.0 Hz), 7.73 (2H, d, J = 8.8 Hz), 7.85 (2H, d, J = 8.8 Hz), 8.15 (1H, s), 10.16 (1H, s), 11.69 (1H, s).

[0480] Example 13. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N- (4-(N-ethylsulfamoyl)phenyl)acetamide (Compound 13) Example 14. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N- methyl-N-(4-(N-methylsulfamoyl)phenyl)acetamide (Compound 14)

[0481]

[0482] Step A. Synthesis of 4-amino-N-ethylbenzenesulfonamide

[0483] Pd / C (50 mg) was added to a solution of N-ethyl-4-nitrobenzenesulfonamide (200 mg, 0.87 mmol, 1.00 equivalent) in EtOH (10 mL) at room temperature. The resulting mixture was stirred at room temperature in H2 (1 atm) for 16 hours. The mixture was filtered and dried to give 4-amino-N-ethylbenzenesulfonamide (170 mg, 97.7%) as a yellow solid, which was used directly in the next step without further purification. [M+H] + = 201.

[0484] Step B. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-ethylaminosulfonyl)phenyl)acetamide (compound 13)

[0485] At room temperature, pyridine (120.10 mg, 1.52 mmol, 4.00 equivalent) and 4-amino-N-ethylbenzenesulfonamide (92.72 mg, 0.38 mmol, 1.00 equivalent) were added to a stirred solution of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetic acid (102 mg, 0.39 mmol, 1.00 equivalent) and HATU (442 mg, 1.16 mmol, 3.00 equivalent) in DMF (6 mL). The resulting mixture was stirred at 40 °C for 24 hours. The DMF was removed, the mixture was extracted with EA, dried over Na2SO4, filtered, and concentrated. The obtained product was purified by preparative HPLC under the following conditions: RP-PREP-1 XBridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 40% B over 10 min; wavelength: 214 nm. The title compound (31 mg, 18.0%) was obtained as a white solid. [M+H] + =444, 1 H NMR (400 MHz, DMSO- d 6): δ 0.96 (3H, t, J = 7.6 Hz), 2.65-2.67 (4H, m), 2.72-2.79 (2H, m), 3.25 (2H, s), 3.93-3.95 (4H, m), 6.62 (1H, t, J = 1.6 Hz), 7.18 (1H, t, J = 2.4 Hz), 7.42 (1H, t, J= 5.6 Hz), 7.73 (2H, d, J = 8.8Hz), 7.86 (2H, d, J = 8.8 Hz), 8.15 (1H, s), 10.17 (1H, s), 11.70 (1H, s).

[0486] Example 14. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N- methyl-N-(4-(N-methylsulfamoyl)phenyl)acetamide (Compound 14) Example 17. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N,N- diethylsulfamoyl)phenyl)acetamide (Compound 15)

[0487]

[0488] Step A. Synthesis of N-methyl-4-(methylamino)benzenesulfonamide

[0489] To a solution of 4-amino-N-methylbenzenesulfonamide (0.20 g, 1.08 mmol, 1.00 equivalent) and paraformaldehyde (0.29 g, 3.23 mmol, 3.00 equivalent) in MeOH (10 mL), 30% MeONa / MeOH (0.58 g, 3.23 mmol, 3.00 equivalent) was added. The mixture was stirred at 65 °C for 16 h. Then, NaBH4 (0.12 g, 3.23 mmol, 3.00 equivalent) was slowly added to the mixture at room temperature, followed by heating to 65 °C for 2 h. LCMS showed the desired product. The mixture was concentrated under vacuum, and water (15 mL) was added to the residue, followed by extraction with EtOAc (3 × 10 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated under vacuum to obtain N-methyl-4-(methylamino)benzenesulfonamide (208 mg, 96%), a yellow oil. [M+H] + =201.

[0490] Step B. Synthesis of 2-chloro-N-methyl-N-(4-(N-methylaminosulfonyl)phenyl)acetamide

[0491] At room temperature, pyridine (119.00 mg, 1.50 mmol, 2.00 equivalent) and DMAP (10.00 mg, 0.08 mmol, 0.10 equivalent) were added to a stirred solution of N-methyl-4-(methylamino)benzenesulfonamide (150.00 mg, 0.75 mmol, 1.00 equivalent) in DCM (10 mL), followed by dropwise addition of a solution of 2-chloroacetyl chloride (85.00 mg, 0.75 mmol, 1.00 equivalent) in DCM (1 mL). The resulting mixture was stirred at room temperature for 1 hour. LCMS showed the reaction was complete. The mixture was diluted with H2O and extracted with DCM. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give crude 2-chloro-N-methyl-N-(4-(N-methylaminosulfonyl)phenyl)acetamide (210 mg, 100%) as a yellow solid. [M+H] + =277.

[0492] Step C. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-methyl-N-(4-(N-methylaminosulfonyl)phenyl)acetamide (compound 14)

[0493] At room temperature, to a solution of 2-chloro-N-methyl-N-(4-(N-methylaminosulfonyl)phenyl)acetamide (100.00 mg, 0.36 mmol, 1.00 equivalent) in EtOH / DMF (8 mL / 5 mL), 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (73.00 mg, 0.36 mmol, 1.00 equivalent), TEA (182.00 mg, 1.80 mmol, 5.00 equivalent), and KI (30.00 mg, 0.18 mmol, 0.50 equivalent) were added. The resulting mixture was stirred at 80 °C for 2 hours. LCMS showed the desired product. The mixture was concentrated under vacuum. The residue was purified by preparative HPLC under the following conditions: column: RP-PREP-5 Xbridge C18 column, 5 μm, 19 × 150 mm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 10% B to 40% B over 11 min; wavelength: 214 nm. The title compound (55 mg, 34%) was given as a white solid. [M+H] + = 444. 1 H NMR (400 MHz, DMSO- d 6): δ 2.42-2.45 (7H, m), 3.15 (2H, s), 3.27 (3H, s), 3.71-3.73 (4H, m), 6.56 (1H, d, J = 1.6 Hz), 7.16 (1H, t, J = 2.8 Hz), 7.49 (1H, s), 7.60 (2H, d, J = 8.4 Hz), 7.80 (2H, d, J = 8.4 Hz), 8.11 (1H, s), 11.67 (1H, s).

[0494] Example 16. Synthesis of 1-(2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetyl)-N- phenylpiperidine-4-carboxamide (Compound 16) Example 17. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- isopropylsulfamoyl)phenyl)acrylamide (Compound 17).

[0495]

[0496] Step A. Synthesis of N,N-diethyl-4-nitrobenzenesulfonamide

[0497] At room temperature, 4-nitrobenzenesulfonyl chloride (400 mg, 1.81 mmol, 1.00 equivalent) was added to a solution of dimethylamine in THF (15 mL, 2 M). The resulting mixture was stirred at room temperature for 2 hours. The mixture was concentrated, diluted with NaHCO3 solution, and extracted with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to give N,N-diethyl-4-nitrobenzenesulfonamide (450 mg, 97%) as a yellow solid. [M+H] + =259.

[0498] Step B. Synthesis of 4-amino-N,N-diethylbenzenesulfonamide

[0499] At room temperature and H2 (1 atm), 10% Pd / C (100 mg) was added to a stirred solution of N,N-diethyl-4-nitrobenzenesulfonamide (450 mg, 1.74 mmol, 1.00 equivalent) in MeOH (15 mL). The resulting mixture was stirred at room temperature for 2 hours. The mixture was filtered and concentrated to give 4-amino-N,N-diethylbenzenesulfonamide (352 mg, 88%) as a yellow solid. [M+H] + =229.

[0500] Step C. Synthesis of 2-chloro-N-(4-(N,N-diethylaminosulfonyl)phenyl)acetamide

[0501] At room temperature, K₂CO₃ (425 mg, 3.08 mmol, 2.00 equivalent) was added to a stirred solution of 4-amino-N,N-diethylbenzenesulfonamide (350 mg, 1.54 mmol, 1.00 equivalent) in DCM (10 mL), followed by 2-chloroacetyl chloride (173 mg, 1.54 mmol, 1.00 equivalent). The resulting mixture was stirred at room temperature for 2 hours. The mixture was diluted with H₂O and extracted with DCM. The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to give 2-chloro-N-(4-(N,N-diethylaminosulfonyl)phenyl)acetamide (325 mg, 70%) as a yellow solid. [M+H] + =305.

[0502] Step D. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N,N-diethylaminosulfonyl)phenyl)acetamide (compound 15)

[0503] At room temperature, 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (217 mg, 1.07 mmol, 1.00 equivalent), TEA (216 mg, 2.14 mmol, 2.00 equivalent), and KI (189 mg, 1.07 mmol, 1.0 equivalent) were added to a solution of 2-chloro-N-(4-(N,N-diethylaminosulfonyl)phenyl)acetamide (325 mg, 1.07 mmol, 1.00 equivalent) in EtOH (10 mL). The resulting mixture was stirred at 80 °C for 4 hours. The resulting solution was evaporated, and the residue was purified by preparative HPLC under the following conditions: column: RP-PREP-5 Xbridge C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 35% B to 65% B over 10 min; wavelength: 214 nm. The title compound (200 mg, 40%) was obtained as a white solid. [M+H] + = 472. 1 H NMR (400 MHz, CDCl3): δ 1.03 (t, J = 6.8 Hz, 6H), 2.64-2.66 (m, 4H), 3.13 (q, J = 7.2 Hz, 4H), 3.25 (2H, s), 3.92-3.95 (m, 4H), 6.62 (q, J = 2.0 Hz, 1H), 7.18 (t,J =2.8 Hz, 1H), 7.74 (d, J = 8.8 Hz, 2H), 7.87 (d, J = 8.8 Hz, 2H), 8.14 (s, 1H), 10.19 (s, 1H), 11.69 (s, 1H).

[0504] Example 18. Synthesis of (R)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- isopropylsulfamoyl)phenyl)propionamide (Compound 18) Example 19. Synthesis of (S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N- isopropylsulfamoyl)phenyl)acrylamide (Compound 19)

[0505]

[0506] Step A. Synthesis of tert-butyl 4-(phenylcarbamoyl)piperidine-1-carboxylate

[0507] To a stirred mixture of 1-(tert-butyloxycarbonyl)piperidin-4-carboxylic acid (2.00 g, 8.73 mmol, 1.00 equivalent) and HATU (3.98 g, 10.47 mmol, 1.20 equivalent) in DCM (30 mL), DIEA (2.25 g, 17.46 mmol, 2.00 equivalent) and aniline (812 mg, 8.73 mmol, 1.00 equivalent) were added. The resulting mixture was stirred at room temperature for 16 hours. After concentration under vacuum, the residue was purified by rapid chromatography (EA / PE = 25%) to give tert-butyl 4-(phenylcarbamoyl)piperidin-1-carboxylic acid (1.20 g, 50%) as a white solid. [M+H] + = 305.

[0508] Step B. Synthesis of N-phenylpiperidine-4-carboxamide (TFA salt)

[0509] To a solution of tert-butyl 4-(phenylcarbamoyl)piperidine-1-carboxylate (1.20 g, 3.90 mmol, 1.00 equivalent) in DCM (12 mL), TFA (3 mL) was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum to give N-phenylpiperidine-4-carboxamide (TFA salt; 1.00 g, 85%) as a yellow oil. [M+H] + =205.

[0510] Step C. Synthesis of 1-(2-chloroacetyl)-N-phenylpiperidine-4-carboxamide

[0511] To a solution of N-phenylpiperidin-4-carboxamide (TFA salt; 1.00 g, 3.90 mmol, 1.00 equivalent) in DCM (20 mL), TEA (799 mg, 7.89 mmol, 2.00 equivalent) and 2-chloroacetyl chloride (529 mg, 4.68 mmol, 1.20 equivalent) were added. The mixture was stirred at room temperature for 2 hours. After concentration under vacuum, the residue was purified by rapid chromatography (EA / PE = 30%) to give 1-(2-chloroacetyl)-N-phenylpiperidin-4-carboxamide (600 mg, 55%) as a white solid. [M+H] + = 281.

[0512] Step D. Synthesis of 1-(2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)acetyl)-N-phenylpiperidin-4-carboxamide (compound 16)

[0513] To a solution of 1-(2-chloroacetyl)-N-phenylpiperidin-4-carboxamide (200 mg, 0.71 mmol, 1.00 equivalent) in EtOH (10 mL), 4-(piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidine (218 mg, 0.78 mmol, 1.10 equivalent), TEA (287 mg, 2.84 mmol, 4.00 equivalent), and KI (5 mg) were added. The mixture was stirred at 80 °C for 16 hours. After concentration under vacuum, the crude product was purified by preparative HPLC under the following conditions (column: XBridge C18 SN. column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 30% B to 55% B over 8 minutes; wavelength: 214 nm), yielding the title compound (100 mg, 32%) as a white solid. [M+H] + =448. 1 HNMR (400 MHz, DMSO- d 6 ): δ 1.44-1.45 (m, 1H), 1.47-1.48 (m, 1H), 1.66-1.68(m, 2H), 2.54-2.67 (m, 6H), 3.04-3.16 (m, 2H), 3.44-3.48 (m, 1H), 3.86-3.87(m, 4H), 4.12-4.17 (m, 1H), 4.39-4.42 (m, 1H), 6.61-6.62 (m, 1H), 7.02 (t, J= 3.2 Hz, 1H), 7.18 (t, J = 2.8 Hz, 1H), 7.26-7.29 (m, 2H), 7.59 (d, J = 8.0Hz, 2H), 8.14 (s, 1H), 9.90 (s, 1H), 11.68 (s, 1H).

[0514] Example 20. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin-1- ylsulfamoyl)phenyl)acrylamide (Compound 20) Example 21. Synthesis of (R)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin- 1-ylsulfamoyl)phenyl)acrylamide (Compound 21)

[0515]

[0516] Step A. Synthesis of N-isopropyl-4-nitrobenzenesulfonamide

[0517] At 0 °C, propan-2-amine (1.26 g, 21.3 mmol, 1.50 equivalent) and DIEA (2.33 g, 28.4 mmol, 2.00 equivalent) were slowly added to a stirred solution of 4-nitrobenzenesulfonyl chloride (3.00 g, 14.2 mmol, 1.00 equivalent) in 30 mL of THF. The resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated, diluted with water, and extracted with EtOAc. The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give N-isopropyl-4-nitrobenzenesulfonamide (1.70 g, 49%) as a creamy white solid. [M+H] + = 245.1.

[0518] Step B. Synthesis of 4-amino-N-isopropylbenzenesulfonamide

[0519] At room temperature and in H2 (1 atm), 10% Pd / C (190 mg) was added to a stirred solution of N-isopropyl-4-nitrobenzenesulfonamide (1.70 g, 6.96 mmol, 1.00 equivalent) in EtOH / THF (25 mL / 5 mL). The resulting mixture was stirred at 60 °C for 3 hours. The mixture was filtered to give 4-amino-N-isopropylbenzenesulfonamide (1.30 g, 87.2%) as a creamy white solid. [M+H] + = 215.1.

[0520] Step C. Synthesis of 2-bromo-N-(4-(N-isopropylaminosulfonyl)phenyl)propionamide

[0521] To a solution of 4-amino-N-isopropylbenzenesulfonamide (280 mg, 1.31 mmol, 1.00 equivalent) and 2-bromopropionic acid (298 mg, 1.96 mmol, 1.50 equivalent) in DMF (10 mL), HATU (597 mg, 1.57 mmol, 1.20 equivalent) and pyridine (310 mg, 3.93 mmol, 3.00 equivalent) were added. The resulting mixture was stirred at room temperature for 16 hours. The mixture was extracted with EtOAc and washed with water. The organic layer was dried over Na2SO4 and concentrated. The mixture was purified by silica gel chromatography to give 2-bromo-N-(4-(N-isopropylaminosulfonyl)phenyl)propionamide (424 mg, 92%) as a white solid. [M+H] + =349.0.

[0522] Step D. Synthesis of tert-butyl 4-(1-((4-(N-isopropylaminosulfonyl)phenyl)amino)-1-oxopropyl-2-yl)piperazine-1-carboxylate

[0523] To a solution of 2-bromo-N-(4-(N-isopropylaminosulfonyl)phenyl)propionamide (424 mg, 1.21 mmol, 1.00 equivalent) and piperazine-1-carboxylic acid tert-butyl ester (270 mg, 1.45 mmol, 1.20 equivalent) in ACN (15 mL), DIEA (315 mg, 2.42 mmol, 2.00 equivalent) was added. The mixture was stirred at 85 °C for 2 hours. The mixture was concentrated. The residue was purified by silica gel chromatography to give 4-(1-oxo-1-((4-(piperidin-1-ylsulfonyl)phenyl)amino)propyl-2-yl)piperazine-1-carboxylic acid tert-butyl ester (498 mg, 90%) as a white solid. [M+H] + = 455.2.

[0524] Step E. Synthesis of N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt)

[0525] To a solution of tert-butyl piperazine-1-carboxylate (120 mg, 0.26 mmol, 1.00 equivalent) in 2 mL of DCM, 1.5 mL of HCl (4 M, in dioxane) was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated to give N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazine-1-yl)propionamide (HCl salt; 110 mg, >100%), as a white solid. [M+H] + = 355.2.

[0526] Step F. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-isopropylaminosulfonyl)phenyl)acrylamide (compound 17)

[0527] DIEA (74 mg, 0.57 mmol, 3.00 equivalent) was added to a solution of N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt) (80 mg, 0.19 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (121 mg, 0.21 mmol, 1.10 equivalent) in 1-butanol (10 mL). The mixture was stirred at 90 °C for 8 hours. The mixture was concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 15% B to 70% B over 10 min; wavelength: 214 nm. The title compound (15.00 mg, 11%) was obtained as a white solid.

[0528] Example 22. Synthesis of (S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin- 1-ylsulfamoyl)phenyl)acrylamide (Compound 22) Example 23. Synthesis of 1-[4-(3-methylpiperidine-1-carbonyl)piperidin-1-yl]-2-(4-[1H- pyrrolo[2,3-d]pyrimidin-4-yl]piperazin-1-yl)ethan-1-one (Compound 23)

[0529]

[0530] Step A. Separation of (R)-4-(1-((4-(N-isopropylaminosulfonyl)phenyl)amino)-1-oxopropyl-2-yl)piperazine-1-carboxylic acid tert-butyl ester by chiral resolution

[0531] 4-(1-((4-(N-isopropylaminosulfonyl)phenyl)amino)-1-oxopropyl-2-yl)piperazine-1-carboxylic acid tert-butyl ester (480 mg) was separated by chiral HPLC to give two enantiomers. The R-enantiomer was obtained in a quantity of 200 mg.

[0532] Step B. Synthesis of (R)-N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt)

[0533] To a solution of (R)-4-(1-((4-(N-isopropylaminosulfonyl)phenyl)amino)-1-oxopropyl-2-yl)piperazin-1-carboxylic acid tert-butyl ester (step A; 120 mg, 0.26 mmol, 1.00 equivalent) in 2 mL of DCM, 1.5 mL of HCl was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated to give (R)-N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)acrylamide (HCl salt; 110 mg, >100%) as a white solid. [M+H] + = 355.2.

[0534] Step C. Synthesis of (R)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-isopropylaminosulfonyl)phenyl)propionamide (compound 18)

[0535] DIEA (74 mg, 0.57 mmol, 3.00 equivalent) was added to a solution of (R)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt) (80 mg, 0.19 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (121 mg, 0.21 mmol, 1.10 equivalent) in 1-butanol (10 mL). The mixture was stirred at 90 °C for 8 hours. The mixture was concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 15% B to 70% B over 10 min; wavelength: 214 nm. (R)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-isopropylaminosulfonyl)phenyl)propionamide (15 mg, 11%) was obtained as a white solid. [M+H] + = 472.2. 1 HNMR (400 MHz, DMSO- d 6 ): δ 0.93 (d, J = 6.8 Hz, 6H), 1.23 (d, J= 7.2 Hz, 3H),2.60-2.75 (m, 4H), 3.14-3.25 (m, 1H), 3.35-3.42 (m, 1H), 3.89-3.94 (m, 4H),6.61 (s, 1H), 7.17 (s, 1H), 7.43 (d, J = 8.0 Hz, 1H), 7.74 (d, J = 8.8 Hz, 2H), 7.85 (d, J = 8.4 Hz, 2H), 8.13 (s, 1H), 10.21 (s, 1H), 11.68 (s, 1H).

[0536] Example 24. Synthesis of N-methyl-N-(propan-2-yl)-1-[2-(4-{1H-pyrrolo[2,3-d]pyrimidin-4- yl}piperazin-1-yl)acetyl]piperidine-4-carboxamide (Compound 24) Example 26. Synthesis of 1-(4-benzylpiperidin-1-yl)-2-(4-{1H-pyrrolo[2,3-d]pyrimidin-4- yl}piperazin-1-yl)ethan-1-one (Compound 26)

[0537]

[0538] Step A. Synthesis of (S)-N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt)

[0539] To a solution of (S)-4-(1-((4-(N-isopropylaminosulfonyl)phenyl)amino)-1-oxopropyl-2-yl)piperazin-1-carboxylic acid tert-butyl ester (Example 13, Step A; 120 mg, 0.26 mmol, 1.00 equivalent) in 2 mL of DCM, 1.5 mL of HCl was added. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated to give (S)-N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt; 70 mg, >100%), as a white solid. [M+H] + = 355.2.

[0540] Step B. Synthesis of (S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(N-isopropylaminosulfonyl)phenyl)acrylamide (compound 19)

[0541] DIEA (78 mg, 0.60 mmol, 3.00 equivalent) was added to a solution of (S)-N-(4-(N-isopropylaminosulfonyl)phenyl)-2-(piperazin-1-yl)propionamide (HCl salt) (70 mg, 0.20 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (31 mg, 0.20 mmol, 1.00 equivalent) in 1-butanol (10 mL). The mixture was stirred at 90 °C for 8 hours. The mixture was concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 15% B to 70% B over 10 min; wavelength: 214 nm. The title compound (18 mg, 19%) was obtained as a white solid. [M+H] + =472.2. 1 H NMR (400 MHz, DMSO- d 6 ): δ 0.93 (d, J = 6.4 Hz, 6H), 1.23 (d, J =6.8Hz, 3H), 2.59-2.74 (m, 4H), 3.14-3.26 (m, 1H), 3.34-3.42 (m, 1H), 3.90-3.94 (m, 4H), 6.05 (s, 1H), 7.17 (s, 1H), 7.73(d, J = 8.8 Hz, 2H), 7.85 (d, J = 8.8 Hz, 2H), 8.13 (s, 1H), 10.21 (s, 1H), 11.66 (s, 1H).

[0542] ​ ​

[0543]

[0544] Step A. Synthesis of 1-((4-nitrophenyl)sulfonyl)piperidine

[0545] At 0 °C, piperidine (1.15 g, 13.54 mmol, 1.50 equivalent) and DIEA (2.33 g, 18.04 mmol, 2.00 equivalent) were slowly added to a stirred solution of 4-nitrobenzenesulfonyl chloride (2.00 g, 9.02 mmol, 1.00 equivalent) in 30 mL of THF. The resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated, diluted with water, and extracted with EtOAc. The organic layer was dried over Na2SO4 and concentrated under reduced pressure to give 1-((4-nitrobenzene)sulfonyl)piperidine (1.97 g, 80.9%) as a creamy white solid. [M+H] + = 271.1.

[0546] Step B. Synthesis of 4-(piperidin-1-ylsulfonyl)aniline

[0547] At room temperature and H2 (1 atm), 10% Pd / C (197 mg) was added to a stirred solution of 1-((4-nitrophenyl)sulfonyl)piperidine (1.97 g, 7.30 mmol, 1.00 equivalent) in EtOH / THF (25 mL / 5 mL). The resulting mixture was stirred at 60 °C for 3 hours. The mixture was filtered and concentrated under vacuum to give 4-(piperidin-1-ylsulfonyl)aniline (1.07 g, 61%) as a creamy white solid. [M+H] + = 241.1.

[0548] Step C. Synthesis of 2-bromo-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide

[0549] To a solution of 4-(piperidin-1-ylsulfonyl)aniline (400 mg, 1.67 mmol, 1.00 equivalent) and 2-bromopropionic acid (306 mg, 2.0 mmol, 1.20 equivalent) in DMF (10 mL), HATU (760 mg, 2.00 mmol, 1.20 equivalent) and pyridine (196 mg, 5.01 mmol, 3.00 equivalent) were added. The resulting mixture was stirred at room temperature for 16 hours. The mixture was extracted with EtOAc and washed with water. The organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The mixture was purified by silica gel chromatography to give 2-bromo-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (500 mg, 80%) as a white solid. [M+H] + = 375.0.

[0550] Step D. Synthesis of tert-butyl piperazine-1-carboxylate

[0551] To a solution of 2-bromo-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (460 mg, 1.23 mmol, 1.00 equivalent) and tert-butyl piperazine-1-carboxylate (274 mg, 1.47 mmol, 1.20 equivalent) in ACN (15 mL), DIEA (316 mg, 2.45 mmol, 2.00 equivalent) was added. The mixture was stirred at 85 °C for 2 hours. LC-MS showed the reaction was complete. The mixture was concentrated, and the residue was purified by silica gel chromatography to give tert-butyl piperazine-1-carboxylate (533 mg, 90%) as a white solid. [M+H] + = 481.2.

[0552] Step E. Synthesis of 2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide

[0553] To a solution of tert-butyl piperazine-1-carboxylate (100 mg, 0.21 mmol, 1.00 equivalent) in 4 mL of DCM, 1.5 mL of HCl (4 M, in dioxane) was added. The mixture was stirred at room temperature for 6 hours. The mixture was concentrated to give 2-(piperazin-1-yl)-N-(4-(piperazin-1-ylsulfonyl)phenyl)propionamide (HCl salt; 80 mg, 91.6%) as a white solid. [M+H] + =381.1.

[0554] Step F. Synthesis of 2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)acrylamide (compound 20)

[0555] To a solution of 2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (80 mg, 0.19 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (121 mg, 0.21 mmol, 1.10 equivalent) in 1-butanol (10 mL), DIEA (74 mg, 0.57 mmol, 3.00 equivalent) was added. The mixture was stirred at 100 °C for 16 h. The mixture was concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 75% B over 10 min; wavelength: 214 nm. The title compound (89 mg, 93%) was obtained as a white solid.

[0556] ​ ​

[0557]

[0558] Step A. Chiral separation of racemic 4-(1-oxo-1-((4-(piperidin-1-ylsulfonyl)phenyl)amino)propyl-2-yl)piperazine-1-carboxylic acid tert-butyl ester

[0559] Racemic tert-butyl 4-(1-oxo-1-((4-(piperidin-1-ylsulfonyl)phenyl)amino)propyl-2-yl)piperazine-1-carboxylate (530 mg) was separated by chiral HPLC to obtain two enantiomers: (S)-enantiomer (234 mg) and (R)-enantiomer (220 mg).

[0560] Step B. Synthesis of (R)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt)

[0561] To a solution of (R)-4-(1-oxo-1-((4-(piperidin-1-ylsulfonyl)phenyl)amino)prop-2-yl)piperazin-1-carboxylic acid tert-butyl ester (100 mg, 0.21 mmol, 1.00 equivalent) in 4 mL of DCM, 1.5 mL of HCl (4 M, in dioxane) was added. The mixture was stirred at room temperature for 6 hours. The mixture was concentrated to give (R)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt; 80 mg, 91.6%), as a white solid. [M+H] + =381.1.

[0562] Step C. Synthesis of (R)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)acrylamide (compound 21)

[0563] To a solution of (R)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt; 80 mg, 0.19 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (121 mg, 0.21 mmol, 1.10 equivalent) in 1-butanol (10 mL), DIEA (74 mg, 0.57 mmol, 3.00 equivalent) was added. The mixture was stirred at 100 °C for 16 hours. The mixture was then concentrated. The residue was purified by preparative HPLC under the following conditions: column: XBridgePrep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 75% B over 10 min; wavelength: 214 nm. The title compound (89 mg, 93%) was obtained as a white solid. [M+H] + =498.2. 1 H NMR (400 MHz, DMSO- d 6 ): δ 1.24 (t, J = 6.8 Hz, 3H),1.34-1.35 (m, 2H), 1.52-1.53 ​​(m, 4H), 2.60-2.73 (m, 4H), 2.85 (t, J = 5.2 Hz,4H), 3.37-3.42 (m, 1H), 3.92 (t, J = 4.8 Hz, 4H), 6.61 (d, J = 2.4 Hz, 1H), 7.18 (t, J = 3.2 Hz, 1H), 7.67 (d, J = 8.8 Hz, 2H), 7.92 (d, J = 8.8 Hz, 2H), 8.14 (s, 1H), 10.28 (s, 1H), 11.68 (s, 1H).

[0564] ​ ​

[0565]

[0566] Step A. Synthesis of (S)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt)

[0567] To a solution of (S)-4-(1-oxo-1-((4-(piperidin-1-ylsulfonyl)phenyl)amino)prop-2-yl)piperazin-1-carboxylic acid tert-butyl ester (Example 26, step A; 100 mg, 0.21 mmol, 1.00 equivalent) in 4 mL of DCM, 1.5 mL of HCl (4 M, in dioxane) was added. The mixture was stirred at room temperature for 6 hours. The mixture was concentrated to give (S)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt; 80 mg, 92%) as a white solid. [M+H] + =381.1.

[0568] Step B. Synthesis of (S)-2-(4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)acrylamide (compound 22)

[0569] DIEA (74 mg, 0.57 mmol, 3.00 equivalent) was added to a solution of (S)-2-(piperazin-1-yl)-N-(4-(piperidin-1-ylsulfonyl)phenyl)propionamide (HCl salt; 80 mg, 0.19 mmol, 1.00 equivalent) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (121 mg, 0.21 mmol, 1.10 equivalent) in 1-butanol (10 mL). The mixture was stirred at 100 °C for 16 hours. LC-MS showed that the reaction was complete. The mixture was concentrated, and the residue was purified by preparative HPLC under the following conditions: XBridge Prep C18 column, 19 × 150 mm, 5 μm; mobile phase A: water (0.1% NH4HCO3), mobile phase B: ACN; flow rate: 15 mL / min; gradient: 20% B to 75% B over 10 min; wavelength: 214 nm. The title compound (81 mg, 84%) was given as a white solid. [M+H] + =498.2. 1 H NMR (400 MHz, DMSO- d 6 ): δ 1.24 (t, J=6.8Hz, 3H), 1.34-1.36 (m, 2H), 1.52-1.53 ​​(m, 4H), 2.60-2.73 (m, 4H), 2.85 (t, J = 5.2 Hz, 4H), 3.37-3.42 (m, 1H), 3.92 (t, J = 4.4 Hz, 4H), 6.61 (d, J =2.0 Hz, 1H), 7.18 (t, J = 3.2 Hz, 1H), 7.67 (d, J = 8.8 Hz, 2H), 7.92 (d, J =8.4 Hz, 2H), 8.14 (s, 1H), 10.28 (s, 1H), 11.68 (s, 1H).

[0570] ​ ​

[0571]

[0572] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (67 mg, 0.244 mmol), 2-chloro-1-[4-(3-methylpiperidin-1-carbonyl)piperidin-1-yl]ethyl-1-one (71 mg, 0.248 mmol), diisopropylamine (122 mg, 1.206 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C185 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% aqueous NH3. The title product was obtained in 59% yield (65.2 mg).

[0573] ​ ​

[0574]

[0575] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (70 mg, 0.254 mmol), 1-(2-chloroacetyl)-N-methyl-N-(propyl-2-yl)piperidine-4-carboxamide (74 mg, 0.284 mmol), diisopropylamine (136 mg, 1.345 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C185 μm 130 Å column (Waters) using a gradient of 20% to 70% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 42% yield (45.3 mg).

[0576] Example 25. Synthesis of N-{4-[(pyrimidin-2-yl)aminosulfonyl]phenyl}-2-(4-{1H-pyrrolo[2,3-d]pyrimidin-4-yl}piperazin-1-yl)acetamide (compound 25)

[0577]

[0578] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (59 mg, 0.214 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-{4-[(pyrimidin-2-yl)aminosulfonyl]phenyl}acetamide (70 mg, 0.215 mmol), diisopropylamine (106 mg, 1.048 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 16% yield (16.8 mg).

[0579] ​ ​

[0580]

[0581] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (72 mg, 0.262 mmol), 1-(4-benzylpiperidin-1-yl)-2-chloroethyl-1-one (67 mg, 0.267 mmol), diisopropylamine (131 mg, 1.295 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 50% to 100% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 49% yield (53.9 mg).

[0582] Example 27. Synthesis of 2-(4-{lH-pyrrolo[2,3-d]pyrimidin-4-yl}piperazin-l- yl)-N-(3-sulfamoylphenyl)acetamide (Compound 27) Example 28. Synthesis of N-phenyl-4-[2-(4-{lH-pyrrolo[2,3-d]pyrimidin-4-yl}piperazin- 1-yl)acetyl]piperazine- 1 -carboxamide (Compound 28)

[0583]

[0584] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (72 mg, 0.262 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-(3-aminosulfonylphenyl)acetamide (69 mg, 0.278 mmol), diisopropylamine (138 mg, 1.365 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 20% to 70% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 33% yield (36 mg).

[0585] Example 29. Synthesis of N-[3-(dimethylsulfamoyl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 29) Example 30. Synthesis of N-[3-(morpholine-4-sulfonyl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 30)

[0586]

[0587] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (65 mg, 0.236 mmol), 4-(2-chloroacetyl)-N-phenylpiperazine-1-carboxamide (71 mg, 0.253 mmol), diisopropylamine (133 mg, 1.315 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 54% yield (57.5 mg).

[0588] Example 31. Synthesis of N-[4-(piperidin- 1 -sulfonyl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 31) Example 32. Synthesis of N-[3-(pyrrolidin- 1 -sulfonyl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 32)

[0589]

[0590] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (66 mg, 0.24 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[3-(dimethylaminosulfonyl)phenyl]acetamide (69 mg, 0.25 mmol), diisopropylamine (121 mg, 1.197 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 41% yield (43.5 mg).

[0591] Example 33. Synthesis of N-[4-(2-phenyldiazen- 1 -yl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 33) Example 34. Synthesis of N-[4-chloro-3-(dimethylsulfamoyl)phenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 34)

[0592]

[0593] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (59 mg, 0.214 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[3-(morpholino-4-sulfonyl)phenyl]acetamide (62 mg, 0.195 mmol), diisopropylamine (109 mg, 1.108 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 36% yield (37.5 mg).

[0594] Example 35. Synthesis of N-[3-(dimethylsulfamoyl)-4-methylphenyl]-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 35) Example 36. Synthesis of N-[3-(dimethylsulfamoyl)phenyl]-2-(4-{lH-pyrrolo[2,3-d]pyrimidin- 4-yl}piperazin- 1 -yl)acrylamide (Compound 36)

[0595]

[0596] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (54 mg, 0.196 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[4-(piperidin-1-sulfonyl)phenyl]acetamide (65 mg, 0.206 mmol), diisopropylamine (102 mg, 1.01 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 50% to 100% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 52% yield (49.8 mg).

[0597] Example 37. Synthesis of N-{4-[methyl(propan-2-yl)sulfamoyl]phenyl}-2-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acetamide (Compound 37) Example 38. Synthesis of 3-(4-{lH-pyrrolo[2,3-d]pyrimidin-4-yl}piperazin-l-yl)-N-(3- sulfamoylphenyl)acrylamide (Compound 38)

[0598]

[0599] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (68 mg, 0.247 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[3-(pyrrolidine-1-sulfonyl)phenyl]acetamide (78 mg, 0.258 mmol), diisopropylamine (129 mg, 1.276 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 38% yield (44.4 mg).

[0600] Example 39. Synthesis of N-[3-(pyrrolidin- 1 -sulfonyl)phenyl]-3-(4-{lH-pyrrolo[2,3- d]pyrimidin-4-yl}piperazin- 1 -yl)acrylamide (Compound 39) Example 40. Synthesis of N-[4-(dimethylsulfamoyl)phenyl]-3-(4-{lH-pyrrolo[2,3-d]pyrimidin- 4-yl}piperazin- 1 -yl)acrylamide (Compound 40)

[0601]

[0602] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (61 mg, 0.222 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[4-(2-phenyldiazepine-1-yl)phenyl]acetamide (67 mg, 0.245 mmol), diisopropylamine (112 mg, 1.108 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 60% to 100% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 68% yield (66 mg).

[0603] ​ ​

[0604]

[0605] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (58 mg, 0.211 mmol), 2-chloro-N-[4-chloro-3-(dimethylaminosulfonyl)phenyl]acetamide (65 mg, 0.21 mmol), diisopropylamine (109 mg, 1.078 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 25% yield (25.1 mg).

[0606] ​ ​

[0607]

[0608] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (60 mg, 0.218 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[3-(dimethylaminosulfonyl)-4-methylphenyl]acetamide (68 mg, 0.234 mmol), diisopropylamine (117 mg, 1.157 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 54% yield (53.8 mg).

[0609] ​ ​

[0610]

[0611] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (66 mg, 0.24 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-[3-(dimethylaminosulfonyl)phenyl]propionamide (74 mg, 0.255 mmol), diisopropylamine (128 mg, 1.266 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 53% yield (57.7 mg).

[0612] ​ ​

[0613]

[0614] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (54 mg, 0.196 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-{4-[methyl(propan-2-yl)aminosulfonyl]phenyl}acetamide (64 mg, 0.21 mmol), diisopropylamine (105 mg, 1.04 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 50% to 100% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 53% yield (49.2 mg).

[0615] ​ ​

[0616]

[0617] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (64 mg, 0.233 mmol) in anhydrous DMSO (approximately 0.5 mL), 3-chloro-N-(3-aminosulfonylphenyl)propionamide (70 mg, 0.267 mmol), diisopropylamine (126 mg, 1.246 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 55% yield (54.7 mg).

[0618] ​ ​

[0619]

[0620] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (55 mg, 0.20 mmol), 3-chloro-N-(3-aminosulfonylphenyl)propionamide (64 mg, 0.202 mmol), diisopropylamine (104 mg, 1.028 mmol), and sodium iodide (0.2 equivalents) were added to anhydrous DMSO (approximately 0.5 mL). The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 50% to 100% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 57% yield (54.9 mg).

[0621] ​ Synthesis of (piperazin- 1 -yl)acrylamide (Compound 40)

[0622]

[0623] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (62 mg, 0.225 mmol) in anhydrous DMSO (approximately 0.5 mL), 3-chloro-N-[4-(dimethylaminosulfonyl)phenyl]propionamide (73 mg, 0.252 mmol), diisopropylamine (122 mg, 1.206 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 40% to 90% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 53% yield (54.3 mg).

[0624] Example 41. Synthesis of N-{[2-(dimethylsulfamoyl)phenyl]methyl}-2-(4-{lH- pyrrolo[2,3-d]pyrimidin-4-yl}piperazin-l-yl)acetamide (Compound 41) Example 41. Synthesis of N-{[2-(dimethylsulfamoyl)phenyl]methyl}-2-(4-{lH- pyrrolo[2,3-d]pyrimidin-4-yl}piperazin-l-yl)acetamide (Compound 41)

[0625]

[0626] To 1-[1H-pyrrolo[2,3-d]pyrimidin-4-yl]piperazine dihydrochloride (66 mg, 0.24 mmol) in anhydrous DMSO (approximately 0.5 mL), 2-chloro-N-{[2-(dimethylaminosulfonyl)phenyl]methyl}acetamide (76 mg, 0.262 mmol), diisopropylamine (129 mg, 1.276 mmol), and sodium iodide (0.2 equivalents) were added. The reaction mixture was sealed and heated at 100 °C for 16 hours. The reaction mixture was then cooled, filtered, and purified by HPLC on an XBridge BEH C18 5 μm 130 Å column (Waters) using a gradient of 30% to 80% 0.1% NH3 / methanol in 0.1% ammonia. The title product was given in 49% yield (54.3 mg).

[0627] Example 42. JAK enzymatic assay

[0628] All enzyme assays were performed according to Eurofins Discovery's KinaseProfiler™ and IC50. 50 Profiler™ service was performed. The test compound stock solution (10 mM, in DMSO) was incubated at nine different concentrations (half-logarithmic dilution series) with the reaction conditions described below to generate and fit IC50. 50 curve.

[0629] Human JAK-1 (Uniprot ID: P23458) was incubated with 20 mM Tris / HCl pH 7.5, 0.2 mM EDTA, 500 μM GEEPLYWSFPAKKK (SEQ ID NO.: 1) peptide substrate, 10 mM magnesium acetate, and [γ-33P]-ATP (specific activity and concentration as needed). The reaction was initiated by adding the Mg / ATP mixture. After incubation at room temperature for 40 minutes, the reaction was terminated by adding phosphoric acid to a concentration of 0.5%. Aliquots of the reaction solution were then spotted onto filter paper, washed four times with 0.425% phosphoric acid for 4 minutes each time, followed by one wash with methanol, then dried and scintillation counted. The known inhibitor astrococcus was used as a positive control for the assay.

[0630] Human JAK-2 (Uniprot ID: O60674) was incubated with 8 mM MOPS at pH 7.0, 0.2 mM EDTA, 100 μM KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC (SEQ ID NO: 2) peptide substrate, 10 mM magnesium acetate, and [γ-33P]-ATP (specific activity and concentration as needed). The reaction was initiated by adding the Mg / ATP mixture. After incubation at room temperature for 40 minutes, the reaction was terminated by adding phosphoric acid to a concentration of 0.5%. Aliquots of the reaction solution were then spotted onto filter paper, washed four times with 0.425% phosphoric acid for 4 minutes each time, followed by one wash with methanol, dried, and then scintillation counted.

[0631] Human JAK-3 (Uniprot ID: P52333) was incubated with 8 mM MOPS at pH 7.0, 0.2 mM EDTA, 500 μM GEEEEYFELVKKKK (SEQ ID NO: 3) peptide substrate, 10 mM magnesium acetate, and [γ-33P]-ATP (specific activity and concentration as needed). The reaction was initiated by adding the Mg / ATP mixture. After incubation at room temperature for 40 minutes, the reaction was terminated by adding phosphoric acid to a concentration of 0.5%. Aliquots of the reaction solution were then spotted onto filter paper, washed four times with 0.425% phosphoric acid for 4 minutes each time, followed by one wash with methanol, dried, and then scintillation counted.

[0632] Human TYK-2 (Uniprot ID: P29597) was incubated with 8 mM MOPS pH 7.0, 0.2 mM EDTA, 250 µM GMGMEDIYFEFMGGKKK (SEQ ID NO: 4) peptide substrate, 10 mM magnesium acetate, and [γ-33P]-ATP (specific activity and concentration as needed). The reaction was initiated by adding the Mg / ATP mixture. After incubation at room temperature for 40 minutes, the reaction was terminated by adding phosphoric acid to a concentration of 0.5%. Aliquots of the reaction solution were then spotted onto filter paper, washed four times with 0.425% phosphoric acid for 4 minutes each time, followed by one wash with methanol, dried, and then scintillation counted.

[0633] The data is then imported into an internal application, where each data point is represented as inhibition % based on uninhibited and enzyme-free controls. Using this scheme, IC50 values ​​of the compound examples of this disclosure are generated. 50 Results. The inhibitory value (IC50) of each compound on JAK enzyme was calculated. 50 The values ​​are divided into four groups. Table 1 provides the group numbers and the selective IC values ​​of JAK1 relative to JAK2. 50 ratio.

[0634] Table 1. JAK inhibitory activity and selectivity

[0635]

[0636] Affinity (IC) 50 Categories: 1: <250 nM; 2: 250 to 500 nM; 3: 500 to 1000 nM; 4: >1000 nM

[0637] Referring to Table 1, many compounds were observed to have good affinity (IC50). 50 <250 nM), and in many cases, the selectivity for JAK1 is greater than 5-fold or greater than 10-fold relative to JAK2. As mentioned above, the very high homology between the human and canine subtypes (nearly 100% at the active site) is considered to be directly translated into the corresponding canine activity, with human assays being a convenient and readily available alternative method.

[0638] Example 43. In vitro T cell proliferation assay in dogs

[0639] T cell activation plays a crucial role in a variety of inflammatory and autoimmune disorders, as well as asthma, allergies, and pruritus. Since T cell activation can be partially triggered by cytokines that signal through the JAK-STAT pathway, JAK inhibitors may be effective for diseases involving aberrant T cell activation.

[0640] Methods:

[0641] Whole blood was collected from 29 beagles and 23 mixed-breed dogs in heparinized sodium tubes. The whole blood (20 µL) was plated together with 180 µL of culture medium (RPMI 1640, Gibco #21870-076, containing 1% heat-inactivated fetal bovine serum, Gibco #10082-39; 292 µg / ml L-glutamine, Gibco #250030-081; 100 µg / ml penicillin and 100 µg / ml streptomycin, Gibco #15140-122) in 96-well plates (Costar 3598) containing a mediator control or test compound (0.001 to 10 µM), concanavalin A (ConA; 1 µg / ml, Sigma C5275), and canine interleukin-2 (IL-2; 50 ng / ml, R&D Systems 1815-CL / CF). Wells containing whole blood, medium with a mediator control, and free of ConA or IL-2 were used as background controls. Plates were incubated at 37°C for 48 hours. 0.4 µCi of tritium-labeled thymidine (PerkinElmer, NetO27A-005MC) was added to each well, and the plates were incubated for another 20 hours. The plates were frozen, then thawed, washed, and filtered using a Brandel MLR-96 cell collector and a pre-wetted filter membrane (Wallac 1205-401, PerkinElmer). The filter membrane was dried at 60°C for one hour (Precision 16EG convection oven) and placed in a filter sample bag (Wallac 1205-411, PerkinElmer) containing 10 mL of scintillation solution (Wallac 1205-440, PerkinElmer). The sealed filter membranes were counted using an LKB Wallac1205 Betaplate liquid scintillation counter. Data was collected using the Gterm Betaplate program vl.l (Wallac copyright, 1989-1990) and converted to inhibition percentages. The data was then graphically displayed as inhibition percentages using GraphPad Prism 4.0, and IC was fitted using point-to-point analysis. 50 curve.

[0642] Example 44. Cell proliferation inhibition assay

[0643] Feline cell lines

[0644] MYA-I and FETJ are feline T lymphoblast cell lines obtained from ATCC (Manassas, VA). These cells were cultured in RPMI 1640 complete medium supplemented with 10% FBS in a humidified incubator containing 5% CO2 at 37°C.

[0645] Ex vivo canine lymphoma node tissue

[0646] Malignant lymph nodes were surgically removed by a veterinarian and placed in transport medium (Advanced RPMI 1640 complete medium supplemented with 10% fetal bovine serum (FBS), 100 U / mL penicillin, 100 μg / mL streptomycin, and 0.25 μg / mL amphotericin B (Invitrogen / Gibco®)). Within 24 hours post-removal, the lymph nodes were processed by mincing them into small pieces and passing them through a tissue sieve. The cell suspension was centrifuged at 200 × g, the supernatant was removed, and the cell pellet was resuspended in NH4Cl solution at room temperature for 10 minutes. The cell suspension was then precipitated by centrifugation; NH4Cl was removed, the pellet was washed once with Hanks' balanced salt solution (HBSS), and then resuspended in proliferation medium (Advanced RPMI complete medium, 1% FBS, 50 nM 2-mercaptoethanol, 100 U / mL penicillin, 100 μg / mL streptomycin, and 0.25 μg / mL amphotericin B). The cell suspension was then passed through a 100 µm nylon cell filter (BD-Falcon) and counted using a hemocytometer. Cells were then placed in a separate proliferation medium supplemented with 0.005% pansorbin. ® Cultured in proliferation medium containing heat-inactivated, formalin-fixed Staphylococcus aureus (SAC) cells (Calbiochem) and 10 ng / mL canine IL-2 (R&D Systems), or proliferation medium supplemented with 125 ng / mL concanavalin A (Sigma) and 125 ng / mL lipopolysaccharide (LPS; Calbiochem).

[0647] Cells cultured in the above culture medium will be at a rate of 1 × 10⁻⁶. 3 Cells / well (cat cell line) or 2 × 10⁻⁶ 5 Cells / well (lymph node cells) were seeded at a density in 96-well Costa plates (Corning) and exposed to different concentrations of test compounds for up to 5 days at 37°C in a humidified incubator containing 5% CO2. CellTiter 96 was used according to the manufacturer's instructions. ®The effect of the aqueous non-radioactive cell proliferation assay (Promega) on proliferation was determined. Typically, proliferation is measured indirectly using soluble tetrazolium salt (MTS) and an electron coupling agent. The bioreduction of MTS to formazan, a product soluble in tissue culture medium, was monitored at absorbance of 490 nM on a Spectramax microplate reader using Softmax Pro 4.6 software (Molecular Devices). Data were graphically displayed as a percentage relative to the DMSO control using GraphPad Prism 4.00, and the IC50 was fitted using a non-linear regression model with an S-shaped dose-response. 50 curve.

[0648] Example 45. Reduction of flea-associated pruritus and dermatitis assay

[0649] Flea-associated pruritus and dermatitis are common skin conditions in dogs. Itching is one of the most severe clinical symptoms associated with flea-associated dermatitis, and persistent scratching, rubbing of the face, and biting of the paws can lead to various skin changes such as erythema, edema, hair loss, lichenification, and hyperpigmentation. Flea-associated pruritus and dermatitis can be induced experimentally. In these models, inflammatory cells and cytokines have been shown to mediate immune responses to allergens. Therefore, JAK inhibitors that inhibit signal transduction of pruritogenic and pro-inflammatory cytokine receptors may be effective in suppressing, alleviating, or minimizing flea-associated pruritus and dermatitis.

[0650] Study design

[0651] Fourteen days before starting medication, use approximately 100 uneaten adult cat fleas (Ctenopharynx catenella) Ctenocephalides felis Twenty-eight mixed-breed male and female dogs weighing 5–35 kg and older than one year were infected with fleas, and each dog was then reinfected with 30 fleas every four days throughout the study period. Seven days prior to administration, 24 dogs were randomly assigned to three different treatment groups based on their Visual Analogue Scale (VAS) scores for skin lesions: placebo, 0.5 mg / kg test compound, or 0.25 mg / kg test compound. Treatment was administered orally twice daily for 28 days, and pruritus behavior, erythema, and skin lesions were assessed during the study. Pruritus behavior was recorded by placing the dogs in a video-enabled enclosure and recording their activities over four hours. Pruritus activity was quantified by determining how many seconds the dogs scratched. Skin lesions were recorded by acquiring images of the abdominal and groin areas and were graded for severity according to the VAS.

[0652] Many modifications and other embodiments of the invention will arise to those skilled in the art from the teachings presented in the foregoing description. Therefore, it should be understood that the invention is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims. Although specific terms are used herein, they are used in a general and descriptive sense only and not for limiting purposes.

Claims

1. A compound having a structure according to Formula I: or a pharmaceutically acceptable salt or solvate thereof, wherein: R1is H or optionally substituted C1-C3alkyl; n = 0 or 1; R2is H or CH3; R3is C1-C4alkyl, phenyl, wherein: m is 0 or 1; X is H, F, CI, OCH3, or CH3; R4is H or CH3; R5is H, C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, or 2-pyrimidinyl; or R4and R5together with the included N atom form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally includes one additional heteroatom selected from N and O; or wherein: R6is H or C1-C4alkyl; R7is H, C1-C4alkyl, phenyl; or R6and R7together with the included N atom form a 6-membered ring, which can optionally be substituted with C1-C4alkyl, and wherein the 6-membered ring optionally includes one additional N atom. (I)， 2. The compound of claim 1, wherein: R2is H; and R3is:

3. The compound of claim 2, wherein n is 1 and R1is H.

4. The compound of claim 1, wherein R3is:

5. The compound of claim 4, wherein m is 0.

6. The compound of claim 5, wherein R4and R5are each H. , or , 7. The compound of claim 6, selected from:

8. The compound of claim 4, wherein m is 1.

9. The compound of claim 8, wherein R4and R5are each CH3.

10. The compound of claim 9, having the structure 11. The compound of claim 4, wherein: R1is H; R2is H; R4is H; and R5is selected from C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidinyl.

12. The compound of claim 4, wherein: R1is CH3; R2is H; R4is H; and R5is selected from C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidinyl. R2and R3together with the included N atom form a 6-membered ring, which is optionally substituted by C1-C4alkyl, C7to C12aralkyl or -C(O)-NR6R7, 12 aralkyl or -C(O)-NR6R7, 13. The compound of claim 11 or 12, selected from:

14. The compound of claim 4, wherein: R1is H or CH3; R2is H; and R4and R5are each independently selected from C1-C4alkyl, C3-C5cycloalkyl, CH2CH2OCH3, and 2-pyrimidinyl.

15. The compound of claim 14, selected from:

16. The compound of claim 4, wherein: R1is H or CH3; R2is H; and R4and R5together with the included N atom form a 4-, 5-, or 6-membered ring, wherein the 6-membered ring optionally includes one additional heteroatom selected from N and O.

17. The compound of claim 16, selected from:

18. The compound of claim 1, which is 19. The compound of claim 1, which is 。 21. The compound of claim 20, having a structure according to Formula II: ​ 。 ​ ​ ​ and . ​ ​ ​ 。 ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ ​ 。 ​ ​ ​ ​ ​ 。 ​ ​ ​ ​ ​ 。 ​ , , or . ​ , , or .

20. The compound of claim 1, wherein R2 and R3 together with the included N atom form a 6-membered ring, which is optionally substituted with C1-C4 alkyl, C7 to C 12 aralkyl or -C(O)-NR6R7. ​ (I), (II), wherein Z is C or N.

22. The compound of claim 21 having a structure selected from: 。 23. The compound of claim 20 having a structure: 。 24. A method for treating an allergic reaction, allergic dermatitis, atopic dermatitis, eczema, or pruritus in a mammal comprising administering to a mammal in need thereof a therapeutically effective amount of a compound according to any one of claims 1-23.

25. The method of claim 24, wherein the mammal is a companion animal.

26. The method of claim 25, wherein the companion animal is a dog.

27. The method of any one of claims 24-26, wherein the compound of Formula I is administered orally, parenterally, or topically.

28. The method of any one of claims 24-27, wherein the compound of Formula I is administered orally once daily.

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