BR112019016949A2 - ANTI-INFLAMMATORY COMPOSITIONS THAT UNDERSTAND IRAK AND JAK INHIBITORS - Google Patents

Abstract

the present invention describes compositions comprising compounds according to formula i: wherein r1, r2 and cy are as defined herein, and a second compound which has a jak inhibitory activity. the present invention relates to compositions, methods for their production, pharmaceutical compositions that comprise them and treatment methods that use them for the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection , diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of il6 or interferons through administration of the compound of the invention.

Description

Invention Patent Descriptive Report for ANTI-INFLAMMATORY COMPOSITIONS THAT UNDERSTAND IRAK AND JAK INHIBITORS.

FIELD OF THE INVENTION

[1] The present invention relates to compositions useful in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving cartilage turnover, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 cartilage or interferons. In a particular aspect, the composition comprises a first compound that has IRAK inhibitory activity and a second compound that has JAK inhibitory activity. The present invention also provides pharmaceutical compositions comprising the combination of the invention and methods for the prophylaxis and / or treatment of diseases, including inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases that involve impairment of cartilage renewal , congenital cartilaginous malformations and / or diseases associated with hypersecretion of IL6 or interferons by administering the combination of the invention.

BACKGROUND OF THE INVENTION

[2] Kinases are involved in many essential processes of cell physiology, for example, protein phosphorylation. In particular, proteins and lipid kinases are involved in cell activation, growth, differentiation and survival. Protein kinases can be divided between those that preferentially phosphorylate tyrosine residues and those that preferentially phosphorylate serine and / or threonine residues.

[3] Over the years, kinases have become very important targets for the development of anti-inflammatory drugs.

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2/154 inflammatory (Cohen, 2009). In particular, kinases associated with the interleukin-1 receptor (IRAK) and, more particularly, IRAK-4, have been identified as playing a role in inflammation and autoimmune diseases (Ringwood and Li, 2008; Wang etaL, 2009).

[4] IRAKs are expressed in many cell types and mediate signals from several cell receptors, including interleukin-1 (IL1) and toll-like receptors (TLRs). In the IRAK family, 4 members were identified, namely IRAK 1-4 and IRAK-4 (Wang et aL, 2009), the most recent member of the family, represents an attractive therapeutic target (Li et aL, 2002). In fact, IRAK-4 is believed to be the main protein kinase activated downstream of the IL-1 and TLRs receptor (except TLR3), initiating signaling via rapid activation of IRAK-1 and IRAK-2, leading to responses innate immune systems. In addition, other interleukins, such as IL-18 and IL-33, are dependent on IRAK-4 for signaling. As such, diseases for which these cytokines are involved in the pathogenic process (eg fibrosis (Li et aL, 2014; McHedlidze et aL, 2013; Rankin et aL, 2010) and atopic dermatitis (Salimi et aL, 2013)) are potential target diseases for treatment with IRAK-4 inhibitors.

[5] In mice expressing an inactive mutant instead of wild type IRAK-4, complete resistance to septic shock triggered by various TLR agonists is observed, as well as a poor response to IL-1. In addition, mice expressing an inactive mutant rather than wild type IRAK-4 are partially protected in several models of autoimmune diseases, such as rheumatoid arthritis (Koziczak-Holbro et al, 2009) and multiple sclerosis (Staschke et aL, 2009) . Interestingly, it has been shown that the serum of patients with rheumatoid arthritis and systemic lupus erythematosus activates plasmacytoid dendritic cells in an IRAK4-dependent manner (Chiang et al, 2011). Finally, pyogenic bacterial infection re

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3/154 current has been observed in children suffering from genetic defects that lead to inactivation of IRAK-4. Since these pyogenic infections are not seen in adults with IRAK-4 inactivating mutations, the IRAK-4 signaling system appears to be redundant for certain aspects of innate immunity in adults.

[6] Deregulation of signaling components of the innate immune system is also increasingly recognized as an important factor in the onset and progression of cancer (Rhyasen and Starczynowski, 2015). In fact, there is evidence that IL-1 plays a direct role in growth, angiogenesis, invasion, drug resistance and tumor cell metastasis (Carmi etal., 2013; Vidal-Vanaclocha et al., 2000). In addition, TLRs are involved in a multiplicity of pro-tumor responses, depending on the context of the tumor cell. As essential mediators of the IL-1 receptor and signaling to TLRs, kinases in the IRAK family represent promising targets for cancer drugs. In addition, several types of cancer have been shown to depend on activated forms of MYD88, an adapter molecule downstream of TLR and IL-1 R which activates IRAK-4. Activating mutations of MYD88 have been identified, for example, in diffuse large B-cell lymphomas (DLBCL) (Ngo et al., 2011) and in Waldenstrom's macroglobulinemia (Treon et al., 2012.). Another report supports the role of IRAK-4 in the field of oncology, acute T-cell lymphoblastic leukemia (T-ALL) in particular (Li etal., 2015). Pharmacological inhibition of IRAK-4 has been shown to increase the sensitivity of TALL to chemotherapeutic agents.

[7] IL-33 has been shown to play a role in the development of fibrotic and allergic diseases, asthma and atopic dermatitis in particular (Nabe, 2014). Since this cytokine signals via an IRAK-4 dependent pathway (Kroeger et al., 2009), these diseases

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4/154 ça may also represent a target for IRAK-4 inhibitors.

[8] Finally, it has been shown that several autoinflammatory diseases are dependent on IL-1 activity and, consequently, biological products that block IL-1 have some benefit for these patients. Gout, juvenile idiopathic arthritis, Muckle-Wells disease, familial Mediterranean fever, Behçet's disease, Still's disease beginning in adulthood are examples of these autoinflammatory diseases (Dinarello etal., 2012).

[9] Inhibition of cytokine signaling with small molecules can help in consequently reducing a disease in immune-inflammatory diseases (Sundberg etal., 2014). In particular, cytokines can play a role in defending organisms against pathogens and infections. However, when developing new therapies for immunoinflammatory diseases, it is crucial, on the one hand, to select a target involved in a pathway that can be inhibited without compromising adaptive and / or innate immune responses, since simultaneous inhibition of multiple response pathways cytokines can excessively weaken the immune system. However, the selectivity of drugs by kinases is difficult to achieve (Bain et at, 2003; Fabian et al., 2005), however, it is highly desirable to avoid associated side effects, particularly in the context of chronic treatments (Broekman etal., 2011; Dy and Adjei, 2013; Force and Kolaja, 2011).

[10] In particular, it has recently been shown that the concomitant use of an agent that blocks IL-1 (anakinra) and a TNF-α blocker (Etanercept) has resulted in an increased risk of infection and neutropenia (Genovese et al., 2003, EMEA public statement, EMEA / 31631/02, February 5, 2003). This discovery highlights that selectivity is a crucial element in the development of new drugs and, therefore, it would be desirable to develop compounds capable of selectively modulating a signaling pathway.

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5/154 without affecting others, in particular compounds capable of selectively modulating the IL-1 response without affecting TNF-a signaling pathways.

[11] Janus kinases (JAKs) are cytoplasmic tyrosine kinases that transduce cytokine signaling from receptors on the membrane to STAT transcription factors. Four members of the JAK family are described, JAK1, JAK2, JAK3 and TYK2. When binding the cytokine to its receptor, members of the JAK family self-and / or transfosphorylate each other, followed by phosphorylation of STATs which then migrate to the nucleus to modulate transcription. JAK-STAT intracellular signal transduction serves interferons, most interleukins, as well as a variety of cytokines and endocrine factors, such as EPO, TPO, GH, OSM, LIF, CNTF, GMCSF and PRL (Vainchenker W. etal. (2008)).

[12] The combination of genetic models and research into small molecule inhibitors of JAK revealed the therapeutic potential of several JAKs. JAK3 is validated by genetic studies with mice and humans as an immunosuppression target (O'Shea J. et al. (2004)). JAK3 inhibitors have been successfully introduced in clinical development, initially for rejection of organ transplants, but later also for other immunoinflammatory indications, such as rheumatoid arthritis (RA), psoriasis and Crohn's disease (http://clinicaltrials.gov /).

[13] TYK2 is a potential target for immunoinflammatory diseases, being validated by genetic studies in humans and knock-out studies with mice (Levy D. and Loomis C. (2007)).

[14] JAK1 is a new target in the area of immunoinflammatory disease. JAK1 heterodimerizes with other JAKs to transduce pro-inflammatory signaling triggered by cytokines. Therefore, it is expected

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6/154 that inhibition of JAK1 and / or other JAKs is of therapeutic benefit for a number of inflammatory diseases, as well as for other diseases triggered by JAK-mediated signal transduction.

[15] Current therapies are unsatisfactory and, therefore, the need remains to identify additional compounds with reduced side effects that can be used in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection. , diseases involving impairment of cartilage renewal, congenital malformations of the cartilage and / or diseases associated with hypersecretion of IL6 or interferons.

BRIEF DESCRIPTION OF THE FIGURES

[16] Figure 1 describes the clinical results in an illustrative Compound 1 ASD therapeutic model at 1 mg / kg bid (solid triangles), 3 mg / kg bid (crosses), 10 mg / kg bid (asterisks) and 30 mg / kg bid (solid circles) compared to the vehicle (solid diamonds) during the period from day 31 to day 46.

[17] Figure 2 describes the clinical results in an ASD therapeutic model for illustrative Compound XXb at 7.5 mg / kg bid (solid triangles) and 15 mg / kg bid (inclined crosses) and Compound 1 (30 mg / kg bid) + Compound XXb (15 mg / kg bid) (solid circles) compared to the vehicle (solid diamonds) from day 31 to day 41.

SUMMARY OF THE INVENTION

[18] The present invention is based on the identification of the composition of the invention useful in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, cartilage malformations congenital and / or diseases associated with hypersecretion of IL6 or interferons. In a particular aspect, the composition comprises a first

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7/154 ro compound that has IRAK inhibitory activity and a second compound that has JAK inhibitory activity.

[19] The present invention also provides pharmaceutical compositions that comprise the combination of the invention and methods for the prophylaxis and / or treatment of diseases, including inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons by administering the combination of the invention.

[20] In a particular aspect, the JAK inhibitor has JAK1 inhibitory activity. In a more particular embodiment, the JAK inhibitor is a selective JAK1 inhibitor.

[21] In another particular aspect, the IRAK inhibitor has IRAK4 inhibitory activity. In a more particular embodiment, the IRAK inhibitor is a selective IRAK4 inhibitor.

[22] Therefore, in a first aspect of the invention, a composition of the invention is provided which comprises:

[23] a compound according to Formula I:

CN

Que where:

[24] Cy is

[25] - C3-7 monocyclic cycloalkyl optionally substituted by one or more independently selected R ³ , or

[26] - monocyclic heterocycloalkyl of 4 to 7 elements that

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8/154 comprises one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more R ³ independently selected;

[27] R ¹ is

[28] - H,

[29] --SO3H,

[30] - -P (= O) (OH) ₂ ,

[31] - C1-4 alkyl,

[32] - -C (= O) - (monocyclic heterocycloalkyl of 4 to 7 elements comprising one or two heteroatoms independently selected from N, S and O) or

[33] - -C (= O) C1-6 alkyl, C1-6 alkyl which is optionally substituted by one or more independently selected R ⁴ groups;

[34] R ² is H or C1-4 alkyl;

[35] each R ³ is independently selected from:

[36] - OH,

[37] - = 0,

[38] - halo, and

[39] - C1-4 alkyl;

[40] each R ⁴ is independently selected from:

[41] --NR ^5a R ^5b ,

[42] - -C (= O) OH,

[43] - 4- to 7-element monocyclic heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more C1-4 alkyl independently selected and

[44] - -NHC (= O) -Ci-4 alkyl-NH ₂ ; and

[45] R ^5a and R ^5b are independently H or C1-4 alkyl;

[46] or a salt or solvate or 0 salt of a pharmaceutical solvate

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9/154 of the same; and

[47] b) a second compound that has a JAK inhibitory activity.

[48] In a particular aspect, the compositions of the invention are provided for use in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, malformations congenital cartilage and / or diseases associated with hypersecretion of IL6 or interferons.

[49] In one aspect, the combination of the invention can inhibit members of the IRAK kinase family and, more particularly, IRAK-4.

[50] In one aspect, the combination of the invention can inhibit members of the JAK kinase family and, more particularly, JAK1.

[51] In another aspect, the combination of the invention may show selectivity for IRAK-4 and JAK1, which can result in improved safety and reduced off-target side effects. In a particular aspect, the combination of the invention can be selective inhibitors of IL-1.

[52] In yet another aspect of the invention, it has also been unexpectedly demonstrated that combinations of the invention exhibit increased effectiveness compared to each individual member. In a particular aspect, the combination of the invention can show a synergy which, in turn, can allow a reduced dosage of each component of the composition of the invention. This can be advantageous to avoid taking unnecessary amounts of drug, while maintaining efficacy and thus reducing the risk of drug-related adverse events.

[53] In another aspect, the present invention provides pharmaceutical compositions that comprise a composition of the invention and a pharmaceutical carrier, excipient or diluent. In one aspect

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In particular, the pharmaceutical composition may further comprise therapeutically active ingredients suitable for use in combination with the compounds of the invention. In a more particular aspect, the additional therapeutically active ingredient is an agent for the treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons.

[54] In addition, the compositions of the invention useful in the pharmaceutical compositions and treatment methods described herein are pharmaceutically acceptable as prepared and used.

[55] In another aspect of the invention, the present invention provides a method of treating a mammal, in particular humans, affected by a condition selected from those listed here and, in particular, inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons, a method which comprises administering an effective amount of the pharmaceutical composition or combinations of the invention as described here.

[56] The present invention also provides pharmaceutical compositions comprising a composition of the invention and a pharmaceutical carrier, excipient or diluent suitable for use in medicine. In a particular aspect, the pharmaceutical composition is for use in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital and / or cartilage malformations disease

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11/154 children associated with hypersecretion of IL6 or interferons.

[57] In additional aspects, the present invention provides methods for synthesizing the compounds of the compositions of the invention, with synthetic protocols and representative pathways described hereinafter.

[58] Other objectives and advantages will become apparent to those skilled in the art from a consideration of the detailed description below.

[59] It should be understood that the compounds of the invention can be metabolized to yield biologically active metabolites.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[60] The following terms are intended to have the meanings given below and are useful for understanding the description and intended scope of the present invention.

[61] When describing the invention, which may include compounds, pharmaceutical compositions containing such compounds and methods of using such compounds and compositions, the following terms, if present, have the following meanings, unless otherwise indicated. It should also be understood that, when described here, any of the portions defined below can be replaced by a variety of substituents and that the respective definitions are intended to include such substituted portions within their scope, as shown below. Unless otherwise indicated, the term replaced must be defined as described below. It should also be understood that the terms groups and radicals can be considered interchangeable when used here.

[62] Articles one and one can be used here to refer to one or more of one (that is, at least one) of the grammatical objects in the article. For example, an analogue means an analogue

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12/154 or more than one analog.

[63] Alkyl means straight or branched aliphatic hydrocarbon that has the specified number of carbon atoms. Particular alkyl groups have 1 to 6 carbon atoms or 1 to 4 carbon atoms. Branched means that one or more alkyl groups, such as methyl, ethyl or propyl, are attached to a linear alkyl chain. Particular alkyl groups are methyl (-CH3), ethyl (-CH2-CH3), npropyl (-CH2-CH2-CH3), isopropyl (-CH (CH3) 2), n-butyl (-CH2-CH2CH2-CH3), tert-butyl (-CH ₂ -C (CH ₃ ) ₃ ), sec-butyl (-CH2-CH (CH ₃ ) 2), n-pentyl (-CH2-CH2-CH2-CH2-CH3), n-hexyl (- CH2-CH2-CH2-CH2-CH2CH ₃ ) and 1,2-dimethylbutyl (-CHCH3) -C (CH ₃ ) H2-CH2-CH ₃ ). Particular alkyl groups have between 1 and 4 carbon atoms.

[64] Alkenyl refers to monovalent olefinically (in) saturated hydrocarbon groups with the specified number of carbon atoms. The particular alkenyl has 2 to 8 carbon atoms and, more particularly, from 2 to 6 carbon atoms, which can be straight or branched and which has at least 1 and, particularly, from 1 to 2 olefinic unsaturation sites. Particular alkenyl groups include ethylene (-CH = CH2), n-propenyl (-ChbChkChk), isopropenyl (-C (CH3) = CH2) and so on.

[65] Alkylene refers to divalent alkene radical groups having 0 number of specified carbon atoms, in particular having 1 to 6 carbon atoms and, more particularly, from 1 to 4 carbon atoms, which can be straight or branched chain. This term is exemplified by groups such as methylene (-CH2), ethylene (-CH2-CH2-) or -CH (CH3) - and so on.

[66] Alkylene refers to divalent alkaline radical groups that have 0 number of carbon atoms and 0 number of specified triple bonds, in particular 2 to 6 carbon atoms and, more particularly, 2 to 4 carbon atoms, which can be in jail

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13/154 straight or branched. This term is exemplified by groups such as -C = C-, -CH ₂ -ChC- and -C (CH ₃ ) H-ChCH-.

[67] Aloxy refers to the O-alkyl group, where the alkyl group has the specified number of carbon atoms. In particular, the term refers to the -0-0-6 alkyl group. Particular alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, npentoxy, n-hexoxy and 1,2-dimethylbutoxy. Particular alkoxy groups are lower alkoxy, that is, with between 1 and 6 carbon atoms. Other particular alkoxy groups have between 1 and 4 carbon atoms.

[68] Amino refers to the radical -NH2.

[69] Arila refers to a monovalent aromatic hydrocarbon group derived by the removal of a hydrogen atom from a single carbon atom from a parental aromatic ring system. In particular, aryl refers to a fused aromatic, monocyclic or polycyclic ring structure, with the number of atoms specified in the ring. Specifically, the term includes groups comprising from 6 to 10 elements in the ring. Particular aryl groups include phenyl and naphthyl.

[70] Cycloalkyl refers to a non-aromatic, monocyclic, fused polycyclic, bridged polycyclic or spirocyclic ring structure, with the number of atoms specified in the ring. A cycloalkyl can have from 3 to 12 carbon atoms, in particular from 3 to 10 and, more particularly, from 3 to 7 carbon atoms. Such cycloalkyl groups include, by way of example, single ring structures, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

[71] Cyan refers to the radical -CN.

[72] Halo or halogen refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I). Particular halo groups are fluorine or chlorine.

[73] Straight, when used to describe a compound or

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14/154 a group present in a compound, means that one or more carbon atoms in the compound or group have been replaced by a nitrogen, oxygen or sulfur heteroatom. Hetero can be applied to any of the hydrocarbyl groups described above, such as alkyl, for example, heteroalkyl, cycloalkyl, for example, heterocycloalkyl, aryl, for example, heteroaryl, and so on that has from 1 to 4 and, particularly, from 1 to 3 hetero atoms, more typically 1 or 2 hetero atoms, for example, a single hetero atom.

[74] Heteroaryl means a fused aromatic, monocyclic or polycyclic ring structure, which includes one or more heteroatoms independently selected from O, N and S and the number of atoms specified in the ring. In particular, the aromatic ring structure can have from 5 to 9 elements in the ring. The heteroaryl group can be, for example, a monocyclic ring of five elements or six elements or a fused bicyclic structure formed from fused rings of five and six elements or two fused rings of six elements or, by way of another example, two fused rings of five elements. Each ring can contain up to four heteroatoms typically selected from nitrogen, sulfur and oxygen. Typically, the heteroaryl ring will contain up to 4 heteroatoms, more typically up to 3 heteroatoms, more usually up to 2, for example, a single heteroatom. In one embodiment, the heteroaryl ring contains at least one nitrogen atom in the ring. The nitrogen atoms in the heteroaryl rings can be basic, as in the case of an imidazole or pyridine, or essentially non-basic, as in the case of an indole or pyrrole nitrogen. In general, the number of basic nitrogen atoms present in the heteroaryl group, including any amino group substituting the ring, will be less than five.

[75] Examples of five-membered monocyclic heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thiophe groups

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15/154 nila, imidazolyl, furazanil, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl.

[76] Examples of six-membered monocyclic heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.

[77] Particular examples of bicyclic heteroaryl groups that contain a five-element ring fused to another five-element ring include, but are not limited to, imidazothiazolyl and imidazoimidazolyl.

[78] Particular examples of bicyclic heteroaryl groups containing a six-element ring fused to a five-element ring include, but are not limited to, benzofuranyl, benzothiophenyl, benzoimidazolyl, benzoxazolyl, isobenzoxazolyl, benzisoxazolyl, benzothiazolyl, benzoisothiazolyl, isobenzolyl, isobenzolyl, isobenzol , indolizinyl, purinyl (e.g., adenine, guanine), indazolyl, pyrazolopyrimidinyl, triazolopyrimidinyl and pyrazolopyridinyl.

[79] Particular examples of bicyclic heteroaryl groups containing two fused six-element rings include, but are not limited to, quinolinyl, isoquinolinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinolinyl, phthalazinyl, naphthyridinyl and pteridinyl groups. Particular heteroaryl groups are those derived from thiophenyl, pyrrolyl, benzothiophenyl, benzofuranyl, indolyl, pyridinyl, quinolinyl, imidazolyl, oxazolyl and pyrazinyl.

[80] Examples of representative heteroarilas include the following:

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[81] where each Y is selected from> C = O, NH, O and S.

[82] Heterocycloalkyl means a fully saturated, monocyclic, fused polycyclic, spirocyclic or bridged polycyclic ring structure that includes one or more heteroatoms selected independently from O, N and S and the number of atoms specified in the ring. The heterocycloalkyl ring structure can have from 4 to 12 elements in the ring, in particular from 4 to 10 elements in the ring and, more particularly, from 4 to 7 elements in the ring. Each ring can contain up to four heteroatoms typically selected from nitrogen, sulfur and oxygen. Typically, the heterocycloalkyl ring will contain up to 4 hetero atoms, more typically up to 3 hetero atoms, more usually up to 2, for example, a single hetero atom. Examples of heterocyclic rings include, but are not limited to, azetidinyl, oxetanyl, tietanyl, pyrrolidinyl (for example, 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl), tetrahydrofuranyl (for example, 1-tetrahydrofuranyl, 2-tetrahydrofuranyl and 3-tetrahydrofuranyl), tetrahydrothiophenyl (eg 1-tetrahydrothiophenyl, 2-tetrahydrothiophenyl and 3-tetrahydrothiophenyl), piperidinyl (eg 1-piperidinyl, 2-piperidinyl, 3-piperidinyl and 4-piperidinyl and 4-piperidinyl and 4 ), tetrahydropyranyl (e.g., 4-tetrahydropyranyl), tetrahydrothiopyranyl (e.g., 4-tetrahydrothiopyranyl), morpholinyl, thiomorpholinyl, dioxanyl, or piperazinyl.

[83] As used here, the term heterocycloalkenyl means a heterocycloalkyl that comprises at least one double bond. Particular examples of heterocycloalkenyl groups are shown in the following illustrative examples:

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[84] where each W is selected from CH2, NH, O and S; each Y is selected from NH, O, C (= 0), SO2 and S; and each Z is selected from N or CH.

[85] Particular examples of monocyclic rings are shown in the following illustrative examples:

[86] where each W and Y are independently selected from -CH2-, -NH-, -O- and -S-.

[87] Particular examples of fused bicyclic rings are shown in the following illustrative examples:

[88] where each W and Y are independently selected from -CH2-, -NH-, -O- and -S-.

[89] Particular examples of bridged bicyclic rings are shown in the following illustrative examples:

[90] where each W and Y are independently selected from -CH2-, -NH-, -O- and -S- and each Z is selected from N or CH.

[91] Particular examples of spirocyclic rings are shown in the following illustrative examples:

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[92] where each Y is selected from -CH2-, -NH-, -O- and -S- ..

[93] Hydroxyl 'refers to the radical -OH.

[94] Oxo refers to the radical = 0.

[95] Substituted refers to a group in which one or more hydrogen atoms are each independently replaced by the same or different substituent (s).

[96] Sulfo or sulfonic acid refers to a radical such as -SO3H.

[97] Thiol refers to the -SH group.

[98] As used here, the term substituted by one or more refers to one to four substituents. In one embodiment, it refers to one to three substituents. In other embodiments, it refers to one or two substituents. In yet another modality, it refers to a substituent.

[99] Thioalkoxy refers to the -S- alkyl group in which the alkyl group has the specified number of carbon atoms. In particular, the term refers to the -S-C1-6 alkyl group. Particular thioalkoxy groups are thiomethoxy, thioethoxy, n-thiopropoxy, isothiopropoxy, n-thiobutoxy, tert-thiobutoxy, sec-thiobutoxy, n-thiopentoxy, n-thio-hexoxy and 1,2-dimethylthiobutoxy. Particular thioalkoxy groups are lower thioalkoxy, that is, with between 1 and 6 carbon atoms. Other particular alkoxy groups have between 1 and 4 carbon atoms.

[100] Those skilled in the organic synthesis technique will recognize that the minimum number of heteroatoms in a chemically viable stable heterocyclic ring, whether aromatic or non-aromatic, is determined by the size of the ring, the degree of unsaturation and the valence of the heteroatoms. In general, a heterocyclic ring can have one to four hetero atoms, as long as the heteroaromatic ring is chemical

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[101] Pharmaceutically acceptable means approved or approved by a federal or state government regulatory agency or the corresponding agency in countries other than the United States or listed in the U.S. Pharmacopoeia or other pharmacopoeia generally recognized for use in animals, more particularly, in humans.

[102] Pharmaceutically acceptable salt refers to a salt of a compound of the invention that is pharmaceutically acceptable and that has the desired pharmacological activity of the parent compound. In particular, these salts are non-toxic and can be acid addition salts and inorganic or organic base addition salts. Specifically, these salts include: (1) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and so on; or formed with organic acids, such as acetic acid, propionic acid, hexanoic acid, cyclopentanopropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanesulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid , hydroxinaftoic acid, salicylic acid, stearic acid, muconic acid and so on; or (2) salts formed when an acidic proton in the parent compound is replaced by a metal ion, for example, an alkali metal ion, a

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20/154 alkaline earth metal ion or aluminum ion; or coordinated with an organic base, such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and so on. Salts also include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium and so on; and, when the compound contains basic functionality, salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and so on. The term pharmaceutically acceptable cation refers to an acceptable cationic counterion of an acidic functional group. Such cations are exemplified by cations of sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium and so on.

[103] Pharmaceutically acceptable vehicle refers to a diluent, adjuvant, excipient or carrier with which a compound of the invention is administered.

[104] Prodrugs refers to compounds, including derivatives of the compounds of the invention, that have cleavable groups and become, through solvolysis or under physiological conditions, the compounds of the invention that are pharmaceutically active in vivo. Such examples include, but are not limited to, choline ester derivatives and so on, N-alkylmorpholine esters and so on.

[105] Solvate refers to forms of the compound that are associated with a solvent, usually through a solvolysis reaction. This physical association includes hydrogen bonding. Conventional solvents include water, EtOH, acetic acid and so on. The compounds of the invention can be prepared, for example, in crystalline form and can be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include stoichiometric solvates and non-stoichiometric solvates. In certain cases, the solvate will be capable of isolation, for

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21/154 example, when one or more solvent molecules are incorporated into the crystalline network of the crystalline solid. Solvate comprises solution and isolatable phase solvates. Representative solvates include hydrates, ethanolates and methanolates.

[106] Individual includes human beings. The terms human, patient and individual are used interchangeably here.

[107] Effective amount means the amount of a compound of the invention that, when administered to an individual to treat a disease, is sufficient to effect such treatment for the disease. The effective amount may vary depending on the compound, the disease and its severity, and the age, weight, etc., of the individual to be treated.

[108] Prevention or prevention refers to a reduction in the risk of acquiring or developing a disease or disorder (that is, preventing at least one of the clinical symptoms of the disease from developing) in an individual who may be exposed to an agent causing the disease or predisposed to the disease before the onset of the disease.

[109] The term prophylaxis is related to prevention and refers to a measure or procedure aimed at preventing, rather than treating or curing a disease. Non-limiting examples of prophylactic measures may include administration of vaccines; the administration of low molecular weight heparin to hospitalized patients at risk of thrombosis due, for example, to immobilization; and administration of an antimalarial agent, such as chloroquine, before a visit to a geographical region where malaria is endemic or the risk of contracting malaria is high.

[110] Treating or treating any disease or disorder refers, in one embodiment, to improving the disease or disorder (that is, stopping the disease or reducing the manifestation, extent or severity of at least one of the clinical symptoms). In another modality,

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22/154 treatment or treating refers to improving at least one physical parameter that may not be discernible by the individual. In yet another modality, treatment or treating refers to the modulation of the disease or disorder, whether physically (for example, stabilization of a discernible symptom), physiologically (for example, stabilization of a physical parameter) or both. In another modality, treatment or treating refers to slowing the progression of the disease.

[111] As used here, the term inflammatory disease (s) refers to the group of conditions that includes rheumatoid arthritis, osteoarthritis, juvenile idiopathic arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, allergic airway disease (for example , asthma, rhinitis), chronic obstructive pulmonary disease (COPD), inflammatory bowel diseases (eg, Crohn's disease, ulcerative colitis), sarcoidosis, endotoxin-induced disease states (eg, complications after bypass surgery or endotoxin states that contribute, for example, to chronic heart failure) and cartilage-related diseases such as joints. In particular, the term refers to rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, osteoarthritis, allergic airway disease (eg, asthma), sarcoidosis, chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. More particularly, the term refers to rheumatoid arthritis, sarcoidosis, psoriatic arthritis, ankylosing spondylitis and inflammatory bowel diseases.

[112] As used here, the term autoimmune disease (s) refers to the group of diseases that includes obstructive airway disease, including conditions such as COPD, asthma (eg, intrinsic asthma, extrinsic asthma, asthma by dust, childhood asthma), particularly chronic or confirmed asthma (eg, late asthma and airway hyper-reactivity), bronchitis, including bronchial asthma, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus, nephritis

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23/154 membranous lupus, dermatomyositis, Sjogren's syndrome, multiple sclerosis, psoriasis, dry eye disease, type I diabetes and associated complications, atopic eczema (atopic dermatitis), thyroiditis (Hashimoto's thyroiditis and autoimmune), contact dermatitis and additional eczematous dermatitis, inflammatory bowel disease (eg Crohn's disease and ulcerative colitis), atherosclerosis and amyotrophic lateral sclerosis, alopecia areata, vitiligo. In particular, the term refers to COPD, asthma, cutaneous lupus erythematosus, membranous lupus nephritis, alopecia areata, vitiligo, type I diabetes mellitus and inflammatory bowel disease.

[113] As used here, the term proliferative disease (s) refers to conditions such as cancer (for example, uterine leiomyosarcoma or prostate cancer), myeloproliferative disorders (for example, polycythemia vera, essential thrombocytosis and myelofibrosis) , leukemia (for example, acute myeloid leukemia, acute and chronic lymphoblastic leukemia), multiple myeloma, psoriasis, restenosis, scleroderma or fibrosis. In particular, the term refers to cancer, leukemia, multiple myeloma and psoriasis.

[114] As used here, the term cancer refers to a malignant or benign growth of cells in the skin or organs of the body, for example, but without limitation, breast, prostate, lung, kidney, pancreas, stomach or intestine. A cancer tends to infiltrate adjacent tissue and spread (metastasize) to distant organs, for example, bone, liver, lung or brain. As used here, the term cancer includes both metastatic tumor-type cells (such as, but not limited to, melanoma, lymphoma, leukemia, fibrosarcoma, rhabdomyosarcoma and mastocytoma) and tissue-type carcinoma (such as, but not limited to, colorectal cancer, prostate cancer, small cell lung cancer and non-small cell lung cancer, breast cancer, pancreatic cancer, cancer

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24/154 bladder cancer, kidney cancer, gastric cancer, glioblastoma, primary liver cancer, ovarian cancer, prostate cancer and uterine leiomyosarcoma). In particular, the term cancer refers to acute lymphoblastic leukemia, acute myeloid leukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytomas, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer ( osteosarcoma and malignant fibrous histiocytoma), brainstem glioma, brain and spinal cord tumors, breast cancer, bronchial tumors, Burkitt's lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniofaring , lymphoma, embryonic tumors, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, Ewing's sarcoma family, eye tumors, eye cancer, retinoblastoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (gastrointestinal stromal tumor) GIST), gastrointestinal stromal cell tumor, germ cell tumor, glioma, cell leukemia hair, head and neck cancer, hepatocellular (liver) cancer, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell endocrine tumors), Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer , leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, hairy cell leukemia, liver cancer, non-small cell lung cancer, large cell lung cancer, Burkitt's lymphoma, cutaneous T lymphoma , non-Hodgkin's lymphoma, lymphoma, Waldenstrom's macroglobulinemia, medulloblastoma, meduloepithelioma, melanoma, mesothelioma, mouth cancer, chronic myeloid leukemia, myeloid leukemia, multiple myeloma, nasopharyngeal cancer, neuroblastoma, non-small cell lung cancer , oropharyngeal cancer, osteos

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25/154 sarcoma, malignant bone fibrous histiocytoma, ovarian cancer, epithelial ovarian cancer, ovarian germ cell tumor, low potential malignant ovarian tumor, pancreatic cancer, papillomatosis, parathyroid cancer, penis cancer, pharyngeal cancer, intermediate differentiating parenchymal tumors, pineoblastoma and primitive supratentorial neuroectodermal tumors, pituitary tumor, plasma cell neoplasia / multiple myeloma, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, kidney (kidney) cancer, kidney (renal) cells , rhabdomyosarcoma, salivary gland cancer, sarcoma, Sezary syndrome, skin cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer, T cell lymphoma, testicular cancer, cancer of throat, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, uterine sarcoma, vagi cancer nal, vulvar cancer, Waldenstrom's macroglobulinemia and Wilms' tumor.

[115] As used here, the term leukemia refers to neoplastic diseases of the blood and blood-forming organs. Such diseases can cause dysfunction of the bone marrow and the immune system, which makes the host highly susceptible to infections and bleeding. In particular, the term leukemia refers to acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) and chronic lymphoblastic leukemia (CLL).

[116] As used here, the term allergic disease (s) refers to the group of conditions characterized by a hypersensitivity disorder of the immune system, including allergic airway disease (e.g., asthma, rhinitis), sinusitis, eczema and hives, as well as food allergies or allergies to insect venom.

[117] As used here, the term asthma refers to any disorder of the lungs characterized by variations in gas flow

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26/154 pulmonary associated with airway constriction of any cause (intrinsic, extrinsic or both; allergic or non-allergic). The term asthma can be used with one or more adjectives to indicate the cause.

[118] As used here, the term transplant rejection refers to the acute or chronic rejection of allografts of solid cells, tissues or organs, for example, pancreatic islets, stem cells, bone marrow, skin, muscle, corneal tissue, tissue neuronal, heart, lung, combined heart-lung disease, kidney, liver, intestine, pancreas, trachea or esophagus or graft versus host disease.

[119] As used here, the term diseases involving impaired cartilage renewal includes conditions such as osteoarthritis, psoriatic arthritis, juvenile rheumatoid arthritis, gouty arthritis, septic or infectious arthritis, reactive arthritis, reflex sympathetic dystrophy, algodystrophy, Tietze syndrome or costal chondricte, fibromyalgia, osteochondritis, neurogenic or neuropathic arthritis, arthropathy, endemic forms of arthritis, such as endemic deforming osteoarthritis, Mseleni disease and Handigodu disease; degeneration resulting from fibromyalgia, systemic lupus erythematosus, scleroderma and ankylosing spondylitis.

[120] As used here, the term congenital cartilaginous malformation (s) includes conditions such as hereditary chondrolysis, chondrodysplasias and pseudochondrodysplasias, in particular, but without limitations, microtia, annotation, metaphyseal chondrodysplasia and related disorders.

[121] As used here, the term disease (s) associated with IL6 hypersecretion includes conditions such as Castleman's disease, multiple myeloma, psoriasis, Kaposi's sarcoma and / or mesangial proliferative glomerulonephritis.

[122] As used here, the term disease (s) associated with

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27/154 interferon hypersecretion includes conditions such as systemic and cutaneous lupus erythematosus, lupus nephritis, dermatomyositis, Sjögren's syndrome, psoriasis, rheumatoid arthritis.

[123] Composition (s) of the invention and equivalent expressions are intended to encompass compounds of the Formula (s) as described herein, an expression that includes pharmaceutically acceptable salts, and solvates, for example, hydrates, and solvates of pharmaceutically acceptable salts, when the context permits. Likewise, the reference to intermediaries, whether or not they are claimed in themselves, is intended to cover their salts and solvates, when the context allows it.

[124] When tracks are mentioned here, for example, but without limitations, C1-8alkyl, the quote for a track should be considered as a representation of each member of that track.

[125] Other derivatives of the compounds of the present invention have activity in both their acid and acid forms, however, in the acid-sensitive form, they often offer advantages of solubility, tissue compatibility or delayed release in a mammal's organism (Bundgard , H, 1985)). Prodrugs include acid derivatives well known to those skilled in the art such as, for example, esters prepared by reacting the parent acid with a suitable alcohol or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendent in the compounds of the present invention are particularly useful prodrugs. In some cases, it is desirable to prepare double ester-type prodrugs, such as (acyloxy) -alkyl esters or ((alkoxycarbonyl) oxy) -alkyl esters. Particularly, such prodrugs are the esters of C1-8 alkyl, C2-8 alkenyl, optional Ce-io aryl

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28/154 and (Ce-io aryl) - (C1-4 alkyl) compounds of the invention.

[126] As used here, the term isotopic variant refers to a compound that contains unnatural proportions of isotopes in one or more of the atoms that make up that compound. For example, an isotopic variant of a compound can contain one or more non-radioactive isotopes, such as, for example, deuterium ( ² H or D), carbon-13 ( ¹³ C), nitro ( ¹⁵ N) or similar. It should be understood that, in a compound in which such an isotopic substitution is made, the following atoms, when present, may vary so that, for example, any hydrogen atom can be ² H / D, any carbon atom can be ¹³ C or any nitrogen can be ¹⁵ N, and that the presence and positioning of such atoms can be determined within the skill of the art. Likewise, the invention can include the preparation of isotopic variants with radioisotopes, for example, where the resulting compounds can be used for tissue and / or substrate tissue distribution studies. The radioactive isotopes of tritium, that is, ³ H, and carbon-14, that is, ¹⁴ C, are particularly useful for this purpose, taking into account their ease of incorporation and ready means of detection. In addition, compounds that are replaced by positron-emitting isotopes, such as ¹¹ C, ¹⁸ F, ¹⁵ O and ¹³ N, can be prepared and these will be useful in Positron Emission Topography (PET) studies to examine the occupation of receivers on substrates.

[127] It should also be understood that compounds that have the same molecular formula, but differ in the nature or binding sequence of their atoms or the arrangement of their atoms in space, are called isomers. Isomers that differ in the arrangement of their atoms in space are called stereomers.

[128] Stereomers that are not mirror images of each other

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29/154 others are called diastereomers and those that are mirror images not superimposed on each other are called enantiomers. When a compound has an asymmetric center, for example, it is linked to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by Cahn and Prelog's R and S sequencing rules or by the way in which the molecule rotates the polarized light plane and is designated as dextrorotatory or levogiratory (that is, as (+) or (-) isomers, respectively). A chiral compound can exist as an individual enantiomer or as a mixture thereof. A mixture that contains equal proportions of the enantiomers is called a racemic mixture.

[129] Tautomers refer to compounds that are interchangeable forms of a particular compound structure and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures can be in equilibrium through the movement of π electrons and an atom (usually H). For example, enols and ketones are tautomers, as they are quickly interconverted through treatment with acid or base. Another example of tautomerism is the acid and nitro forms of phenylnitromethane, which are also formed by treatment with acid or base.

[130] Tautomeric forms may be relevant for obtaining the ideal chemical reactivity and biological activity of a compound of interest.

[131] The compounds of the invention can have one or more asymmetric centers; such compounds can therefore be produced as individual (R) or (S) stereomers or as mixtures thereof.

[132] Unless otherwise specified, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic

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30/154 cas or not. Methods for determining stereochemistry and separating stereomers are well known in the art.

[133] It will be appreciated that the compounds of the invention can be metabolized to produce biologically active metabolites.

THE INVENTION

[134] The present invention is based on the identification of the composition of the invention, useful in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, malformations congenital cartilage and / or diseases associated with hypersecretion of IL6 or interferons. In a particular aspect, the composition comprises a first compound that has IRAK inhibitory activity and a second compound that has JAK inhibitory activity.

[135] The present invention also provides pharmaceutical compositions comprising the combination of the invention and methods for the prophylaxis and / or treatment of diseases, including inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons by administering the combination of the invention.

[136] In a particular aspect, the JAK inhibitor has JAK1 inhibitory activity. In a more particular embodiment, the JAK inhibitor is a selective JAK1 inhibitor.

[137] In another particular aspect, the IRAK inhibitor has IRAK4 inhibitory activity. In a more particular embodiment, the IRAK inhibitor is a selective IRAK4 inhibitor.

[138] Therefore, in a first aspect of the invention, a composition of the invention is provided which comprises:

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[139] a compound according to Formula I:

CN where:

[140] Cyé

[141] - C3-7 monocyclic cycloalkyl optionally substituted by one or more independently selected R ³ , or

[142] - monocyclic heterocycloalkyl of 4 to 7 elements comprising one or two hetero atoms independently selected from N, S and O, optionally substituted by one or more R ³ independently selected;

[143] R ¹ is

[144] - H,

[145] --SO3H,

[146] - -P (= O) (OH) ₂ ,

[147] - C1-4 alkyl,

[148] - -C (= O) - (monocyclic heterocycloalkyl of 4 to 7 elements comprising one or two heteroatoms independently selected from N, S and O) or

[149] - -C (= O) C1-6 alkyl, C1-6 alkyl which is optionally substituted by one or more independently selected R ⁴ groups;

[150] R ² is H or C1-4 alkyl;

[151] each R ³ is independently selected from:

[152] -OH,

[153] - = O,

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[154] - halo, and

[155] - C1-4 alkyl;

[156] each R ⁴ is independently selected from:

[157] --NR ^5a R ^5b ,

[158] --C (= O) OH,

[159] - 4- to 7-element monocyclic heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more independently selected C1-4 alkyl and

[160] - -NHC (= O) -Ci-4 alkyl-NH ₂ ; and

[161] R ^5a and R ^5b are independently H or C1-4 alkyl;

[162] or a salt or solvate or a pharmaceutically acceptable solvate salt thereof; and

[163] b) a second compound that has a JAK inhibitory activity.

[164] In one embodiment, the composition of the invention comprises the compound according to Formula I, wherein Cy is monocyclic C37cycloalkyl. In a particular embodiment, Cy is cyclohexyl.

[165] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein Cy is C3-7 monocyclic cycloalkyl substituted by one, two or three independently selected R ^3's . In a particular embodiment, Cy is cyclohexyl substituted by one, two or three R ³ independently selected. In another particular embodiment, Cy is C3-7 monocyclic cycloalkyl substituted by one or two R ³ . In a more particular embodiment, Cy is cyclohexyl substituted by one or two independently selected R ³ .

[166] In one embodiment, the composition of the invention comprises a compound according to Formula I, where Cy is heterocycle

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33/154 alkyl of 4 to 7 elements comprising one or two heteroatoms independently selected from N, S and O. In a particular embodiment, Cy is tetrahydropyranyl or tetrahydrothiopyranyl. In a more particular mode, Cy is ⁰ .

[167] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein Cy is a 4- to 7-element monocyclic heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, replaced by one, two or three R ³ independently selected. In another embodiment, Cy is tetrahydrofuran or tetrahydrofuran hidropiranila hidrotiopiranila, each of which is substituted by one, two or three independently selected R ^3. In a particular embodiment, Cy is a 4- to 7-element heterocycloalkyl that comprises one or two heteroatoms independently selected from N, S and O, replaced by one or two R ³ . In another particular embodiment, Cy is tetrahydropyranyl or tetrahydrothiopyranyl hidrotiopiranila, each of which is substituted by one or two independently selected from R ^3. In an ity modali More particularly, Cy is or ^S, each of which is substituted by one or two independently selected from R ^3.

[168] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein each R ³ is selected from OH, = 0, halo and C1-4 alkyl. In a particular mode, each R ³ is selected from OH, = 0, F and -CH3. In a more particular mode, each R ³ is selected from OH and -CH3. In another more particular mode, each R ³ is F. In yet another more particular mode, each R ³ is = 0.

[169] In one embodiment, the composition of the invention comprises

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34/154 endo compound according to Formulas Ila, lib, He, lid, He or Ilf:

Hd He Ilf

[170] where R ¹ and R ² are as described above.

[171] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ¹ is H, -SO ₃ H or -P (= O) (OH) ₂ .

[172] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ¹ is C 1-4 alkyl. In a particular embodiment, R ¹ is -CH3, -CH2CH3 or -CH (CH3) 2. In a more particular modality, R ¹ is -

CH ₃ .

[173] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-llf, wherein R ¹ is -C (= O) - (4- to 7-element heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O). In a particular embodiment, R ¹ is -C (= O) -pyrrolidinyl.

[174] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ¹ is -C (= O) C1-6 alkyl. In a particular modality, R ¹ is

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-C (= 0) C1-6 alkyl, C1-6 alkyl which is selected from -CH3, -CH2CH3, -CH2CH2CH3 or -CH2 (CH (CH ₃ ) ₂ ).

[175] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-llf, wherein R ¹ is -C (= O) C1-6 alkyl, C1-6 alkyl which is replaced by a or more R ⁴ independently selected. In a particular embodiment, R ¹ is -C (= O) C1-6 alkyl, C1-6 alkyl which is selected from -CH3, -CH2CH3, -CH2CH2CH3 or -CH2 (CH (CH3) 2), each one of which is replaced by one or more independently selected R ⁴ . In another particular embodiment, R ¹ is -C (= O) C 1-6 alkyl, C 1-6 alkyl which is replaced by one or two independently selected R ⁴ . In a more particular embodiment, R ¹ is -C (= O) C1-6 alkyl, C1-6 alkyl which is selected from -CH3, -CH2CH3, -CH ₂ CH ₂ CH3 or -CH ₂ (CH ( CH3) ₂ ), each of which is replaced by one or two independently selected R ⁴ .

[176] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-11f and R ⁴ is -NR ^5a R ^5b , wherein each R ^5a and R ^5b is independently H or C1-4 alkyl. In a particular embodiment, each R ^5a and R ^5b is independently H, -CH3 or -CH2CH3. In a more particular embodiment, R ^5a is H and R ^5b is H, -CH3 or -CH2CH3. In an even more particular embodiment, R ⁴ is -NH ₂ , -NHCH3 or -N (CH ₃ ) ₂ .

[177] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ⁴ is -C (= O) OH.

[178] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-llf, wherein R ⁴ is a 4- to 7-element heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more independent C1-4 alkyl

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36/154 selected. In a particular embodiment, R ⁴ is morpholinyl, piperidinyl or piperazinyl, each of which is optionally substituted by one or more independently selected C1-4 alkyl. In another particular embodiment, R ⁴ is 4- to 7-element heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by a C1-4 alkyl. In a more particular embodiment, R ⁴ is 4- to 7-element heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by a -CH3. In another more particular embodiment, R ⁴ is morpholinyl, piperidinyl or piperazinyl, each of which is optionally substituted by one or more -CH3. In an even more particular embodiment, R ⁴ is morpholinyl, piperidinyl or piperazinyl, each of which is optionally substituted by a -CH3.

[179] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ⁴ is -NHC (= O) -Ci-4 alkyl-NH2. In a particular embodiment, R ⁴ is -NHC (= O) -CH2-NH ₂ .

[180] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-llf, wherein R ¹ is -C (= O) CH ₂ NH ₂ , -C (= O) CH ₂ NHCH ₃ , -C (= O) CH ₂ N (CH ₃ ) ₂ , -C (= O) CH ₂ CH ₂ N (CH3) 2, -C (= O) CH (NH ₂ ) CH (CH ₃ ) ₂ ,

-C (= O) CH ₂ CH ₂ C (= O) OH, -C (= O) CH (NH ₂ ) CH ₂ C (= O) OH,

-C (= O) CH (NH ₂ ) CH ₂ CH ₂ C (= O) OH, -C (= O) CH (CH (CH ₃ ) 2) NHC (= O) CH ₂ NH2,

[181] In one embodiment, the composition of the invention comprises a compound according to any of Formulas l-llf, wherein R ² is H.

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[182] In one embodiment, the composition of the invention comprises a compound according to any of Formulas 1-11f, wherein R ² is C 1-4 alkyl. In a particular embodiment, R ² is -CH3.

[183] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein the compound is selected from:

[184] 6- [6- [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3-yl] - nicotinonitrile,

[185] 6- {5- (1,1-dioxo-tetrahydro-2H-thiopyran-4-ylamino) -6- [2- (2-hydroxy-ethoxy) -ethoxy] -imidazo [4,5-b] pyridin-3-yl} -nicotinonitrile,

[186] 6- {6- [2- (2-h idroxy-ethoxy) -ethoxy] -5 - [((cis-1,4) -4-h idroxy-4-methylcyclohexyl) -methyl- amino] -imidazo [4,5-b] pyridin-3-yl} -nicotinonitrile,

[187] 6- {6- [2- (2-methoxy-ethoxy) -ethoxy] -5- [methyl- (tetrahydro-pyran-4-yl) amino] -imidazo [4,5-b] pyridin -3-il} -nicotinonitrile,

[188] 6- [6- [2- (2-methoxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3-yl] - nicotinonitrile,

[189] 6- {5- (3-hydroxy-cyclohexylamino) -6- [2- (2-hydroxy-ethoxy) -ethoxy] imidazo [4,5-b] pyridin-3-yl} -nicotinonitrile,

[190] 6- {5- (4,4-difluoro-cyclohexylamino) -6- [2- (2-hydroxy-ethoxy) ethoxy] -imidazo [4,5-b] pyridin-3-yl} - nicotinonitrile,

[191] ester mono- (2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-

4-ylamino) -3H-imidazo [4,5-b] pyridin-6-yloxy] -ethoxy} -ethyl) sulfuric acid,

[192] 2- {2- [3- (5-cyano-pyridin-2-i) ester -5- (tetrahydro-pyrid-4-ylamino) -3H-imidazo [4, 5-b] (S) -2amino-3-methyl-butyric acid pyridin-6-yloxy] -ethoxy} -ethyl,

[193] 2- {2- [3- (5-cyano-pyridin-2-i) -5- ester (tetrahydro-pyrid-4-ylamino) -3H-imidazo [4, 5-b] (S) -2amino-3-methyl-butyric acid pyridin-6-yloxy] -ethoxy} -ethyl, oxalic acid salt,

[194] 6- [6- [2- (2-hydroxyethoxy) ethoxy] -5 - [[(cis-3,4) -4-hydroxytetra-

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38/154 hydropyran-3-yl] amino] imidazo [4,5-b] pyridin-3-yl] pyridine-3-carbonitrile,

[195] 6- [6- [2- (2-hydroxyethoxy) ethoxy] -5 - [((cis-1,4) -4-h idroxy-4methylcyclohexyl) amino] imidazo [4,5-b] pyridin-3-yl] pyridine-3-carbonitrile,

[196] 6- [5 - [((cis-1,4) -4-hydroxy-4-methyl-cyclohexyl) -methyl-amino] -6 [2- (2-methoxyethoxy) ethoxy] imidazo [4 , 5-b] pyridin-3-yl] pyridine-3-carbonitrile,

[197] 6- [5 - [((cis-1,4) -4-hydroxy-4-methylcyclohexyl) amino] -6- [2- (2methoxyethoxy) ethoxy] imidazo [4,5-b] pyridine -3-yl] pyridine-3-carbonitrile,

[198] 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6- 2- (dimethylamino) acetate il] oxyethoxy] ethyl,

[199] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy 2-aminoacetate] ethyl,

[200] 2- (methylamino) acetate (2S) -pyrrolidine-2-carboxylate of 2 [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [ 4.5b] pyridin-6-yl] oxyethoxy] ethyl,

[201] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy] ethyl,

[202] 2- [2- [3 (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-) [2-aminoacetyl) amino] -3-methyl-butanoate ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl,

[203] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy 2-morpholinoacetate] ethyl,

[204] 2- [2- [3- (5-cyano-2-pyridyl) 2- (4-methylpiperazin-1-yl) acetate -

5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy] ethyl,

[205] 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6- 3- (dimethylamino) propanoate il] oxyethoxy] ethyl,

[206] 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6- 2- (dimethylamino) acetate il] oxyethoxy] ethyl, oxalic acid salt,

[207] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy 2-aminoacetate] ethyl, acid salt

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39/154 oxalic,

[208] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl 2- (methylamino) acetate ] oxyethoxy] ethyl, oxalic acid salt,

[209] 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin (2S) -pyrrolidine-2-carboxylate -6-yl] oxyethoxy] ethyl, oxalic acid salt,

[210] (3S) -3-amino-4- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy] ethoxy] -4-oxobutanoic, hydrochloric acid salt,

[211] (4S) -4-amino-5- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy] ethoxy] -5-oxopentanoic, hydrochloric acid salt,

[212] (2S) -2 - [(2-aminoacetyl) amino] -3-2- [2- [3 (5-cyano-2-pyridyl) -5- (tetrahydropyran-4- ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl, oxalic acid salt,

[213] 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy 2-morpholinoacetate] ethyl, oxalic acid salt,

[214] 2- [2- [3- (5-cyano-2-pyridyl) 5- (tetrahydropyran-4-ylamino) imidazo [4,5- 2- (4-methylpiperazin-1-yl) acetate [4,5- b] pyridin-6-yl] oxyethoxy] ethyl, oxalic acid salt,

[215] 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6- 3- (dimethylamino) propanoate il] oxyethoxy] ethyl, oxalic acid salt, and

[216] 4- [2- [2- [3- (5-cyano-2-pyridine I) -5- acid (tetrahydro-4-ylamino) imidazo [4,5-b] pyridine -6-yl] oxyethoxy] ethoxy] -4-oxo-butanoic.

[217] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein the compound is 6- [6 [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro- piran-4-ylamino) -imidazo [4,5

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b] pyridin-3-yl] -nicotinonitrile.

[218] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein the compound is not

6- [6- [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3-yl] -nicotinonitrile.

[219] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein the compound is 2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetra (S) -2-Amino-3-methyl-butyric acid -hydro-pyran-4-ylamino) -3Himidazo [4,5-b] pyridin-6-yloxy] -ethoxy} -ethyl.

[220] In one embodiment, the composition of the invention comprises a compound according to Formula I, wherein the compound is not 2- {2- [3- (5-cyano-pyridin-2-yl) -5- ( (S) -2-amino-3-methylbutyric acid tetrahydro-pyran-4-ylamino) -3Himidazo [4,5-b] pyridin-6-yloxy] -ethoxy} -ethyl.

[221] In another embodiment, the composition of the invention comprises a compound that has a JAK inhibitory activity. In a particular embodiment, the compound that has a JAK inhibitory activity is a compound described in WO 2010/010190.

[222] In another embodiment, the composition of the invention comprises a compound that has a JAK1 inhibitory activity. In a particular embodiment, the compound that has a JAK1 inhibitory activity is according to Formula XXa or XXb:

, N

THE

N

7 ° = ° o

XXa

XXb

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[223] In a particular embodiment, the composition of the invention comprises:

[224] A compound selected from 6- [6- [2- (2-hydroxyethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) -imidazo [4,5-b] pyridin- 3-yl] nicotinonitrile and 2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4ylamino) -3H-imidazo [4,5-b] pyridin ester (S) -2amino-3-methyl-butyric acid -6-yloxy] -ethoxy} -ethyl; and

[225] A compound selected from compounds according to Formulas XXa and XXb

[226] In a particular embodiment, the composition of the invention comprises:

[227] A compound selected from 6- [6- [2- (2-hydroxyethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) -imidazo [4,5-b] pyridin- 3-yl] nicotinonitrile and 2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4ylamino) -3H-imidazo [4,5-b] pyridin ester (S) -2amino-3-methyl-butyric acid -6-yloxy] -ethoxy} -ethyl; and

[228] The compound according to Formula XXb

[229] In a more particular embodiment, the composition of the invention comprises:

[230] 6- [6- [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3-yl] - nicotinonitrile; and

[231] the compound according to Formula XXb.

[232] In one embodiment, the compound of the composition of the invention is not an isotopic variant.

[233] In one aspect, the compound of the composition of the invention according to any of the modalities described here is present as the free base.

[234] In one aspect, the compound of the composition of the invention according to any of the modalities described here is a pharmaceutically acceptable salt.

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[235] In one aspect, the compound of the composition of the invention according to any of the embodiments described here is a solvate of the compound.

[236] In one aspect, a composition compound of the invention according to any of the embodiments described herein is a solvate of a pharmaceutically acceptable salt of a compound.

[237] Although the groups specified for each modality were generally listed above separately, a compound of the composition of the invention includes one in which several or each of the modalities in the Formula above, as well as other formulas presented here, are selected from a or more specific members or groups designated, respectively, for each variable. Therefore, the present invention is intended to include all combinations of such modalities within its scope.

[238] Although the groups specified for each modality have generally been listed above separately, a compound of the invention can be one for which one or more variables (for example, R groups) are selected from one or more modalities according to any of the Formulas listed above. Therefore, the present invention is intended to include all combinations of variables of any of the modalities described in its scope.

[239] Alternatively, the exclusion of one or more of the specified variables from a group or a modality, or combinations thereof, is also considered by the present invention.

[240] In certain aspects, the present invention provides prodrugs and derivatives of the compounds according to the above formulas. The prodrugs are derived from compounds of the composition of the invention that have metabolically cleavable groups and become, by means of solvolysis or under physiological conditions, the compounds of the

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43/154 invention, which are pharmaceutically active in vivo. Such examples include, but are not limited to, choline ester derivatives and so on, N-alkylmorpholine esters and so on.

[241] Other derivatives of the compounds of the composition of the present invention have activity in their acid and acid derivative forms, but the acid sensitive form often offers advantages of solubility, tissue compatibility or delayed release in a mammal's organism (Bundgard, H., 1985). Prodrugs include acid derivatives well known to those skilled in the art such as, for example, esters prepared by reacting the parent acid with a suitable alcohol or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine , or acid anhydrides or mixed anhydrides. Esters, amides and simple aliphatic or aromatic anhydrides derived from acidic groups pendent in the compounds of the present invention are preferred prodrugs. In some cases, it is desirable to prepare double ester type prodrugs, such as (acyloxy) alkyl esters or ((alkoxycarbonyl) oxy) -alkyl esters. Particularly, such prodrugs are the C1-8 alkyl, C2-8 alkenyl esters, optionally substituted Ce-io aryl and (Ce-io aryl) - (C1-4 alkyl) compounds of the invention.

PHARMACEUTICAL COMPOSITIONS

[242] When used as a medicament, a composition of the invention is typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound of the invention according to Formula I. In general, a composition of the invention is administered in a pharmaceutically effective amount. The amount of compound of the invention actually administered will typically be determined by a

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44/154 physician in the light of the relevant circumstances, including the condition to be treated, the route of administration chosen, the actual compound of the invention administered, the age, weight and response of the individual patient, the severity of the patient's symptoms and so on .

[243] The pharmaceutical compositions of the present invention can be administered via a variety of routes including oral, rectal, transdermal, subcutaneous, intra-articular, intravenous, intramuscular and intranasal. Depending on the intended route of delivery, a compound of the invention is preferably formulated as injectable or oral compositions or as ointments, as lotions or as adhesives for transdermal administration.

[244] Compositions for oral administration can take the form of liquid bulk solutions or suspensions or bulk powders. Most commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term unit dosage forms refers to physically distinct units suitable as unit dosages for humans and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect, in association with a pharmaceutical excipient, vehicle or carrier. appropriate. Typical unit dosage forms include pre-filled ampoules or syringes and pre-measured liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the composition of the invention is, in general, a minor component (from about 0.1 to about 50% by weight or, preferably, from about 1 to about 40% by weight) ), with the remainder being vehicles or carriers and processing aids useful in forming the desired dosage form.

[245] Liquid forms suitable for oral administration can

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45/154 include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispensing agents, coloring, flavoring and so on. Solid forms can include, for example, any of the following ingredients or compounds of the invention of a similar nature: a binder, such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient, such as starch or lactose, a disintegrating agent, such as alginic acid, Primogel or corn starch; a lubricant, such as magnesium stearate; a glidant, such as colloidal silicon dioxide; a sweetening agent, such as sucrose or saccharin; or a flavoring agent, such as peppermint or orange flavoring.

[246] Injectable compositions are typically based on sterile injectable saline or phosphate buffered saline or other injectable vehicles known in the art. As before, the active compounds of the composition of the invention in such compositions are typically a minor component, often being from about 0.05 to 10% by weight, the remainder being the injectable vehicle, and so on.

[247] Transdermal compositions are typically formulated as a topical ointment or cream that contains the active ingredient (s), usually in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from 0.1 to about 10% by weight and, more preferably, from about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredients will typically be combined with a paraffinic or water-miscible ointment base. Alternatively, the active ingredients can be formulated into a cream, for example, with an oil-in-water cream base. Such transdermal formulations are well known in the art and include, in general, ingredients

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46/154 additional tests to increase skin penetration and stability of active ingredients or formulation. All of these known transdermal formulations and ingredients are included within the scope of the present invention.

[248] A composition of the invention can also be administered by a transdermal device. Therefore, transdermal administration can be performed using a reservoir-like patch or porous membrane or a variety of solid matrix.

[249] The components described above for orally administrable, injectable or topically administrable compositions are representative only. Other materials, as well as processing techniques and so on, are presented in Part 8 of Remington's Pharmaceutical Sciences, 17th Edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[250] A composition of the invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[251] The following formulation examples illustrate representative pharmaceutical compositions that can be prepared according to the present invention. The present invention, however, is not limited to the following pharmaceutical compositions.

Formulation 1 - Tablets

[252] A composition of the invention comprising a compound according to Formula I and a JAK inhibitor can be mixed as a dry powder with a dry gelatin binder in an approximate ratio of 1: 2 by weight. A smaller amount of magnesium stearate can be added as a lubricant. The mixture

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47/154 can be molded into 240-270 mg tablets (80-90 mg of the active compound of the invention according to Formula I per tablet) in a tablet press.

Formulation 2 - Capsules

[253] A composition of the invention comprising a compound according to Formula I and a JAK inhibitor can be mixed as a dry powder with a starch diluent in an approximate 1: 1 weight ratio. The mixture can be filled into 250 mg capsules (125 mg of active compound of the invention according to Formula I per capsule).

Formulation 3 - Liquid

[254] A composition of the invention comprising a compound according to Formula I and a JAK inhibitor (125 mg) can be mixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resulting mixture can be mixed, passed through a US 10 mesh sieve and then mixed with a previously prepared solution of microcrystalline cellulose and sodium carboxymethylcellulose (11:89, 50 mg) in water. Sodium benzoate (10 mg), flavoring and coloring can be diluted with water and added with stirring. Sufficient water can then be added with stirring. More sufficient water can then be added to produce a total volume of 5 ml.

Formulation 4 - Tablets

[255] A composition of the invention comprising a compound according to Formula I and a JAK inhibitor can be mixed as a dry powder with a dry gelatin binder in an approximate ratio of 1: 2 by weight. A smaller amount of magnesium stearate can be added as a lubricant. The mixture can be molded into 450-900 mg tablets (150-300 mg of active compound of the invention according to Formula I) in a tablet press.

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Formulation 5 - Injection

[256] A composition of the invention comprising a compound according to Formula I and a JAK inhibitor can be dissolved or suspended in an aqueous injectable medium of sterile buffered saline to a concentration of approximately 5 mg / mL.

Formulation 6 - Topic

[257] Stearyl alcohol (250 g) and white petrolatum (250 g) may be melted at around 75 ^Q C and then a mixture of an invention composition comprising a compound according to Formula I and a JAK inhibitor ( 50 g), methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate (10 g) and propylene glycol (120 g) dissolved in water (about 370 g) can be added and the mixture resultant can be stirred until hard.

TREATMENT METHODS

[258] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in medicine. In a particular embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions comprising a composition of the invention, for use in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases that involve impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons.

[259] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of inflammatory diseases,

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49/154 autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons.

[260] In further aspects of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impaired renewal of cartilage, congenital cartilaginous malformations and / or diseases associated with hypersecretion of IL6 or interferons, methods which comprise administering an effective amount of a composition of the invention or one or more of the pharmaceutical compositions described herein for the treatment or prophylaxis of said condition.

[261] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular modality, the other therapeutic agent is an agent for the treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases involving impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons.

[262] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of inflammatory diseases. In a particular embodiment, inflammatory disease is selected from rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, osteoarthritis, allergic airway disease, sarcoidosis, chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. In a modality

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50/154 more particularly, the inflammatory disease is selected from rheumatoid arthritis, sarcoidosis, psoriatic arthritis, ankylosing spondylitis and inflammatory bowel diseases.

[263] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of inflammatory diseases. In a particular embodiment, inflammatory disease is selected from rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, osteoarthritis, allergic airway disease, sarcoidosis, chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. In a more particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, sarcoidosis, psoriatic arthritis, ankylosing spondylitis and inflammatory bowel diseases.

[264] In further aspects of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by inflammatory diseases, methods which comprise administering an effective amount of a composition of the invention or one or more of the pharmaceutical compositions described here for the treatment or prophylaxis of said condition. In a particular embodiment, inflammatory disease is selected from rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, osteoarthritis, allergic airway disease, sarcoidosis, chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. In a more particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, sarcoidosis, psoriatic arthritis, ankylosing spondylitis and inflammatory bowel diseases.

[265] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular modality, the other

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51/154 therapeutic agent is an agent for the treatment of inflammatory diseases. In a particular embodiment, inflammatory disease is selected from rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, osteoarthritis, allergic airway disease, sarcoidosis, chronic obstructive pulmonary disease (COPD) and inflammatory bowel diseases. In a more particular embodiment, the inflammatory disease is selected from rheumatoid arthritis, sarcoidosis, psoriatic arthritis, ankylosing spondylitis and inflammatory bowel diseases.

[266] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of autoimmune diseases. In a particular embodiment, autoimmune disease is selected from COPD, asthma, cutaneous lupus erythematosus, membranous lupus nephritis, alopecia areata, vitiligo, type I diabetes mellitus and inflammatory bowel disease.

[267] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of autoimmune diseases. In a particular embodiment, autoimmune disease is selected from COPD, asthma, cutaneous lupus erythematosus, membranous lupus nephritis, alopecia areata, vitiligo, type I diabetes mellitus and inflammatory bowel disease.

[268] In further aspects of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by autoimmune diseases, methods which comprise administering an effective amount of a composition of the invention or one or more of the pharmaceutical compositions described here for the treatment or prophylaxis of said condition. In a modali

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52/154 particularity, autoimmune disease is selected from COPD, asthma, cutaneous lupus erythematosus, membranous lupus nephritis, alopecia areata, vitiligo, type I diabetes mellitus and inflammatory bowel disease.

[269] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of autoimmune diseases. In a particular embodiment, autoimmune disease is selected from COPD, asthma, cutaneous lupus erythematosus, membranous lupus nephritis, alopecia areata, vitiligo, type I diabetes mellitus and inflammatory bowel disease.

[270] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of proliferative diseases. In a particular embodiment, the proliferative disease is selected from cancer, leukemia, multiple myeloma and psoriasis.

[271] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of proliferative diseases. In a particular embodiment, the proliferative disease is selected from cancer, leukemia, multiple myeloma and psoriasis.

[272] In further aspects of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by proliferative diseases, methods which comprise administering an effective amount of a composition of the invention or one or more of the pharmaceutical compositions described here for the treatment or prophylaxis of said condition. In a modali

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53/154 particularity, the proliferative disease is selected from cancer, leukemia, multiple myeloma and psoriasis.

[273] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of proliferative diseases. In a particular embodiment, the proliferative disease is selected from cancer, leukemia, multiple myeloma and psoriasis.

[274] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of allergic diseases. In a particular embodiment, the allergic disease is eczema.

[275] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of allergic diseases. In a particular embodiment, the allergic disease is eczema.

[276] In a further aspect of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by allergic diseases, methods which comprise administering an effective amount of a composition of the invention or one or more of the compositions pharmaceuticals described here for the treatment or prophylaxis of said condition. In a particular embodiment, the allergic disease is eczema.

[277] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of allergic diseases. In a particular embodiment, the allergic disease is eczema.

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[278] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of transplant rejection. In a particular embodiment, transplant rejection is graft versus host disease.

[279] In another embodiment, the present invention provides compositions of the invention or pharmaceutical compositions that comprise a composition of the invention for use in the manufacture of a medicament for use in prophylaxis and / or transplant rejection treatment. In a particular embodiment, transplant rejection is graft versus host disease.

[280] In further aspects of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by transplant rejection, methods which comprise administering an effective amount of a composition of the invention or one or more of the compositions pharmaceuticals described here for the treatment or prophylaxis of said condition. In a particular embodiment, transplant rejection is graft versus host disease.

[281] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of transplant rejection. In a particular embodiment, transplant rejection is graft versus host disease.

[282] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of diseases that involve deterioration of cartilage renewal. In a particular modality, the disease that involves

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55/154 impairment of cartilage renewal is ankylosing spondylitis.

[283] In another embodiment, the present invention provides compositions of the invention, or pharmaceutical compositions comprising a composition of the invention, for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of diseases involving impairment of cartilage renewal. . In a particular embodiment, the disease that involves impaired cartilage renewal is ankylosing spondylitis.

[284] In a further aspect of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by diseases involving impairment of cartilage renewal, methods which comprise administering an effective amount of a composition of the invention or one or more of the pharmaceutical compositions described herein for the treatment or prophylaxis of said condition. In a particular embodiment, the disease that involves impaired cartilage renewal is ankylosing spondylitis.

[285] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of diseases that involve impaired cartilage renewal. In a particular modality, the disease involving impairment of cartilage renewal is ankylosing spondylitis.

[286] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of congenital cartilage malformations.

[287] In another embodiment, the present invention provides

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56/154 sections of the invention, or pharmaceutical compositions comprising a composition of the invention, for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of congenital cartilage malformations.

[288] In a further aspect of the treatment method, the present invention provides methods of prophylaxis and / or treatment of a mammal affected by congenital cartilage malformations, methods which comprise administering an effective amount of a composition of the invention or one or more of pharmaceutical compositions described herein for the treatment or prophylaxis of said condition.

[289] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for treating an agent for treating congenital cartilage malformations.

[290] In one embodiment, the present invention provides a composition of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the prophylaxis and / or treatment of diseases associated with hypersecretion of IL6 or interferons.

[291] In another embodiment, the present invention provides compositions of the invention, or pharmaceutical compositions that comprise a composition of the invention, for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of diseases associated with hypersecretion of IL6 or interferons.

[292] In a further aspect of the treatment method, the present invention provides methods of prophylaxis and / or treatment of an affected mammal with diseases associated with hypersecretion of IL6 or interferons, methods which comprise administering an effective amount of a composition of the invention. or one or more of

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57/154 pharmaceutical compositions described herein for the treatment or prophylaxis of said condition.

[293] In one embodiment, the present invention provides pharmaceutical compositions that comprise a composition of the invention and another therapeutic agent. In a particular embodiment, the other therapeutic agent is an agent for the treatment of diseases associated with hypersecretion of IL6 or interferons.

[294] Injection dose levels range from about 0.1 mg / kg / h to at least 10 mg / kg / h, generally from about 1 to about 120 h and especially from 24 to 96 h. A preload bolus of about 0.1 mg / kg to about 10 mg / kg or more can also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 1 g / day for a human patient weighing 40 to 80 kg.

[295] For the prophylaxis and / or treatment of long-term conditions, such as degenerative conditions, the treatment regimen generally extends over many months or years, so oral dosing is preferred for the patient's convenience and tolerance. With oral administration, one to four (1-4) regular daily doses, especially one to three (1-3) regular daily doses, typically one to two (1-2) regular daily doses and, more typically, one (1) regular daily dose are representative regimens. Alternatively, for long-acting drugs, with oral dosing, once every two weeks, once a week and once a day are representative regimens. In particular, the dosage regimen may be every 1-14 days, more particularly 1-10 days, even more particularly 1-7 days and, more particularly, 1-3 days.

[296] Using these dosage standards, each dose provides from about 1 to about 1000 mg of a composition of the invention, with particular doses providing each from about

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58/154 from 10 to about 500 mg, and especially about 30 to about 250 mg.

[297] Transdermal doses are generally selected to provide blood levels similar or lower than those obtained with injectable doses.

[298] When used to prevent the appearance of a condition, a composition of the invention will be administered to a patient at risk of developing the condition, typically under the guidance and supervision of a physician, at the dosage levels described above. Patients at risk of developing a particular condition generally include those who have a family history of the disease or those who have been identified by genetic testing or screening as being particularly susceptible to the development of the condition.

[299] A composition of the invention can be administered as a single active agent or it can be administered in combination with other therapeutic agents, including another compound of the invention, which demonstrates the same therapeutic activity or a similar therapeutic activity and which is determined to be safe and effective for such combined administration. In a specific modality, the co-administration of two (or more) agents allows significantly lower doses of each to be used, thus reducing the observed side effects.

[300] In one embodiment, a composition of the invention or a pharmaceutical composition that comprises a composition of the invention is administered as a medicament. In a specific embodiment, said pharmaceutical composition further comprises another active ingredient.

[301] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or pro

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59/154 phylaxis of a disease that involves inflammation. Particular agents include, but are not limited to, immunoregulatory agents, for example, azathioprine, corticosteroids (for example, prednisolone or dexamethasone), cyclophosphamide, cyclosporine A, tacrolimus, mycophenolate, mofetil, muromonabe-CD3 (OKT3, for example, Orthocolone®) ATG, aspirin, acetaminophen, ibuprofen, naproxen and piroxicame.

[302] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of arthritis (e.g., rheumatoid arthritis). Specific agents include, but are not limited to, analgesics, non-steroidal anti-inflammatory drugs (NSAIDS), steroids, synthetic DMARDs (for example, but not limited to, methotrexate, leflunomide, sulfasalazine, auranofin, sodium aurothiomalate, penicillamine, chloroquine, chloroquine, hydroxy tofacitinib, baricitinib, fostamatinib and cyclosporine) and biological DMARDs (eg, without limitation, infliximab, etanercept, adalimumab, rituximab and abatacept).

[303] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of proliferative disorders. Specific agents include, but are not limited to: methotrexate, leucovorin, adriamycin, prednisone, bleomycin, cyclophosphamide, 5-fluorouracil, paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, doxorubicin, tamoxifen, toremifene, anhydrol, antitrogen, meglolone, anhydrolone; anti-HER2 (eg Herceptin ™), capecitabine, raloxifene hydrochloride, EGFR inhibitors (eg Iressa®, Tarceva ™, Erbitux ™), VEGF inhibitors (eg Avastin ™), proteasome inhibitors (eg Velcade ™), Glivec® and hsp90 inhibitors (for example, 17-AAG). In addition, the composition of the invention comprising a compound according to Formula I and an inhibitor

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60/154 JAK pain can be administered in combination with other therapies including, but not limited to, radiation therapy or surgery. In a specific modality, the proliferative disorder is selected from cancer, myeloproliferative disease or leukemia.

[304] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of autoimmune diseases. Specific agents include, but are not limited to: glucocorticoids, cytostatic agents (for example, purine analogs), alkylating agents (for example, nitrogen mustards (cyclophosphamide), nitrosoureas, platinum compounds of the invention and others), antimetabolites (for example, methotrexate, azathioprine and mercaptopurine), cytotoxic antibiotics (eg dactinomycin anthracyclines, mitomycin C, bleomycin and mitramycin), antibodies (eg anti-CD20, anti-CD25 or anti-CD3 (OTK3) monoclonal antibodies, Atgam® and Thymoglobuline®), cyclosporine, tacrolimus, rapamycin (sirolimus), interferons (eg, IFN-β), TNF-binding proteins (eg, infliximab, etanercept or adalimumab), mycophenolate, fingolimod and myiocin.

[305] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of transplant rejection. Specific agents include, but are not limited to: calcineurin inhibitors (eg cyclosporine or tacrolimus (FK506)), mTOR inhibitors (eg sirolimus, everolimus), antiproliferatives (eg azathioprine, mycophenolic acid), corticosteroids (eg , prednisolone, hydrocortisone), antibodies (for example, IL2Ra anti-receptor monoclonal antibodies, basiliximab, daclizumab), polyclonal antibodies against T cells (eg, anti-thymocyte globulin (ATG), anti-lymphocyte globulin (ALG)).

[306] In one embodiment, a composition of the invention is co

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61/154 administered with another therapeutic agent for the treatment and / or prophylaxis of asthma and / or rhinitis and / or COPD. Particular agents include, but are not limited to: beta2-adrenoceptor agonists (eg salbutamol, levalbuterol, terbutaline and bitolterol), epinephrine (inhalable or tablets), anticholinergics (eg, ipratropium bromide), glucocorticoids (oral or inhaled), long-acting p2-agonists (eg, salmeterol, formoterol, bambuterol and oral sustained release albuterol), combinations of inhaled steroids and long-acting bronchodilators (eg fluticasone / salmeterol, budesonide / formoterol), leukotriene antagonists and synthesis inhibitors (eg, montelukast, zafirlukast and zileuton), mediator release inhibitors (eg, cromoglycate and ketotifen), biological regulators of the IgE response (eg, omalizumab), antihistamines (eg, ceterizine, cinnarizine, fexofenadine) and vasoconstrictors (for example, oxymetazoline, xylometazoline, naphazoline and tramazoline).

[307] In addition, a composition of the invention can be administered in combination with emergency therapies for asthma and / or COPD, such therapies including administration of oxygen or heliox, nebulized salbutamol or terbutaline (optionally combined with an anticholinergic (for example, ipratropium ), systemic steroids (oral or intravenous, for example, prednisone, prednisolone, methylprednisolone, dexamethasone or hydrocortisone), intravenous salbutamol, injected or inhaled nonspecific beta-agonists (eg epinephrine, isoetarin, isoproterenol, isoproterenol, methoxyphenol, nebulized, for example, glycopyrrolate, atropine, ipratropium), methylxanthines (theophylline, aminophylline, bamifiline), inhaled anesthetics that have a bronchodilator effect (eg, isoflurane, halothane, enflurane), ketamine and intravenous magnesium sulfate.

[308] In one embodiment, a composition of the invention is co

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62/154 administered with another therapeutic agent for the treatment and / or prophylaxis of inflammatory bowel disease (IBD). Specific agents include, but are not limited to: glucocorticoids (eg, prednisone, budesonide) synthetic disease-modifying agents, immunomodulating agents (eg, methotrexate, leflunomide, sulfasalazine, mesalazine, azathioprine, 6-mercaptopurine and cyclosporine) and disease-modifying agents (infliximab, adalimumab, rituximab and abatacept).

[309] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of SLE. Particular agents include, but are not limited to: human monoclonal antibodies (belimumab (Benlysta)), disease-modifying antirheumatic drugs (DMARDs), such as antimalarials (eg, plaquenyl, hydroxychloroquine), immunosuppressants (eg, methotrexate and azathioprine), cyclophosphamide and mycophenolic acid, immunosuppressive and analgesic drugs, such as non-steroidal anti-inflammatory drugs, opiates (for example, dextropropoxyphene and co-codamol), opiates (for example, hydrocodone, oxycodone, MS Contin or methadone) and the transdermal patch. fentanyl.

[310] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of psoriasis. Specific agents include, but are not limited to: topical treatments, such as bath solutions, moisturizers, medicated creams and ointments containing coal tar, dithranol (anthralin), corticosteroids, such as deoxymethasone (Topicort ™), fluocinonide, vitamin D3 analogs (eg calcipotriol), argan oil and retinoids (etretinate, acitretin, tazarotene), systemic treatments such as methotrexate, cyclosporine, retinoids, thioguanine, hydroxyurea, sulfasalazine, mycophenolate mofetil, azathiopri

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63/154 na, tacrolimus, fumaric acid esters or biological products, such as Amevive ™, Enbrel ™, Humira ™, Remicade ™, Raptiva ™ and ustequinumab (an IL-12 and IL-23 blocker). In addition, a compound of the invention can be administered in combination with other therapies including, but not limited to, phototherapy or photochemotherapy (e.g., psoralen) and ultraviolet A (PUVA) phototherapy).

[311] In one embodiment, a composition of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of an allergic reaction. Specific agents include, but are not limited to: antihistamines (for example, cetirizine, diphenhydramine, fexofenadine, levocetirizine), glucocorticoids (for example, prednisone, betamethasone, beclomethasone, dexamethasone), epinephrine, theophylline, or antileukletrienes or, for example, β-leukthyltes, or β-leukthyltes. ), anticholinergics and decongestants.

[312] By co-administration is meant any way of administering two or more therapeutic agents to the patient as part of the same treatment regimen, as will be evident to those skilled in the art. Although the two or more agents can be administered simultaneously in a single formulation, that is, as a single pharmaceutical composition, this is not essential. The agents can be administered in different formulations and at different times.

SYNTHETIC CHEMICAL PROCEDURES

General

[313] A compound of the composition of the invention can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions (ie reaction temperatures, times, molar ratios of reagents, solvents, pres

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64/154 sections, etc.) are provided, other process conditions can also be used, unless otherwise specified. The ideal reaction conditions may vary with the particular reagents or solvent used, however, such conditions can be determined by those skilled in the art through routine optimization procedures.

[314] Additionally, as will be evident to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protection group for a particular functional group, as well as suitable conditions for protection and deprotection, are well known in the art. For example, numerous protection groups, and their introduction and removal, are described in T. W. Greene and P. G. M. Wuts, Protecting Groups in Organic Synthesis, Wiley-Blackwell; 4- revised edition (2006) and references cited in it (Wuts and Greene, 2006).

[315] The following methods are presented in detail about the preparation of a compound of the composition of the invention as defined above and comparative examples. A compound of the composition of the invention can be prepared from starting materials and reagents known or commercially available to those skilled in the art of organic synthesis.

[316] All reagents are of commercial quality and are used as received without further purification, unless otherwise specified. Commercially available anhydrous solvents are used for reactions conducted under an inert atmosphere. Reagent grade solvents are used in all other cases, unless otherwise specified. Column chromatography is performed on standard silica (30 - 70 pm). Thin layer chromatography is performed using pre-coated silica gel plates 60 F-254 (thickness 0.25 mm). The ¹ H NMR spectra are recorded on a spectrometer

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65/154 meter Bruker Advance 400 MHz or a 300 MHz Bruker Advance DPX spectrometer. Chemical shifts (δ) for ¹ H NMR spectra are presented in parts per million (ppm) relative to tetramethylsilane (δ 0.00 ) or the peak of the appropriate residual solvent as an internal reference. Multiplicities are provided as singlet (s), doublet (d), triplet (t), quartet (q), quintet (quin), multiplet (m) and broad (br). Electrospray MS spectra are obtained on an LC / MS spectrometer on the Waters platform or with a Waters Acquity UPLC with Waters Acquity PDA detector and SQD mass spectrometer. Columns used: UPLC BEH C18 of 1.7 pm, 2.1 X 5 mm, VanGuard precolumn with UPLC BEH C18 of 1.7 pm, 2.1 X 30 mm or Acquidad UPLC BEH C18 of 1.7 pm, 2.1 X 50 mm. All methods use MeCN / H2O gradients. MeCN and H2O contain 0.1% formic acid or 0.05% NH3. Preparative LCMS: used columns, Waters XBridge Prep C18 of 5 pm, ODB 30 X 100 mm (preparative column) and Waters XBridge C18 of 5 pm, 4.6 mm X 100 mm (analytical column). All methods use MeCN / H2O gradients. MeCN and H2O contain 0.1% formic acid or 0.1% diethylamine.

Table I. List of abbreviations used in the experimental section.

Abbreviation Definition AcOH Acetic Acid ΑΡΜΑ 4-aminophenylmercuric acetate aq. aqueous atm atmosphere BINAP (+/-) - 2,2'-bis (diphenylphosphino) -1,1 '-binaftyl Boc tert-butyloxy-carbonyl br broad signal BSA bovine serum albumin Shut up calculated Cdp compound

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Abbreviation Definition d doublet δ chemical bypass DCM dichloromethane dd double doublets DIPEA diisopropylethylamine DMAP dimethylaminopyridine DMF dimethylformamide DMSO Dimethyl sulfoxide dt double triplets DTT dithiothreitol EDCI A / - (3-dimethylaminopropyl) - / V- ilcarbodiimide EDTA ethylenediaminetetraacetic acid eq. equivalent ES- negative electrospray ES + positive electrospray Et ₂ O diethyl ether EtOAc ethyl acetate EtOH ethanol g grass H hour HPLC high performance liquid chromatography Hz hertz Int intermediate iPrOH isopropanol LiOMe lithium methoxide LiOtBu lithium tert-butoxide m multiplet MeCN acetonitrile

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Abbreviation Definition MeOH methanol mg milligram MgOAc magnesium acetate MHz megahertz min minute mL milliliter mmol millimole mol soft MOPS 3- (A / -morpholino) propanesulfonic acid MS mass spectrometry MW molecular weight N normality NaBH (OAc) ₃ sodium triacetoxyborohydride NaOtBu sodium tert-butylate NBS A / -bromosuccinimide NMR nuclear magnetic resonance Obsvd observed PBS phosphate buffered saline PBST phosphate buffered saline with Tween 20 Pd (OAc) ₂ palladium diacetate Pd / C carbon palladium ppm parts per million q quadruple when quadruple doublets quin quintet rt room temperature s singlet

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Abbreviation Definition He sat down. saturated WITHOUT mean standard error t triplet td trio of doublets TEA triethylamine TFA trifluoroacetic acid THF tetrahydrofuran UPLC ultra-performance liquid chromatography

SYNTHETIC PREPARATION OF THE COMPOUNDS OF THE INVENTION

[317] The profile of the compound of the composition of the invention according to Formula (XXa) has been extensively studied and the data are described in WO 2013/189771 (Varit Klooster etal., 2013).

[318] Similarly, the profile of the compound of the invention according to Formula (XXb) has been extensively studied and the data are described in WO 2010/149769 (Menet and Smits, 2010). The synthesis of the salt and suitable formulations has been described in WO2015 / 117980 and WO2015 / 117981.

Example 1. General synthetic methods

1.1 Overview of synthetic methods

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R1

1.2 General Methods

1.2.1 General Method A

R1 N Cl

AT THE.

AT THE.

R1 N NH

R

[319] To a solution of NaH (2 eq., 60% in mineral oil) in dry THF cooled to 0 ° C is added to the corresponding 6-amino-nicotinonitrile (1.1 to 1.2 eq.). After 30 min at 0 ° C, 2-chloro-3-nitropyridine (1 eq.) Is added and the reaction is stirred at r.t. and monitored by means of UPLC-MS. If the reaction is not completed, the reaction is cooled down again to 0 ° C and more NaH is added, followed by more amine. The reaction mixture is poured into ice water and stirred for 2 h. The precipitate is filtered, washed with H2O and air dried under vacuum to provide the desired compound.

Illustrative synthesis of general method A: 6- (6-chloro-3-nitro-pyridin-2-ylamino) -nicotinonitrile (Int 1)

NH.

AT THE.

CN

[320] To a solution of NaH (2.07 g, 51.81 mmol, 2 eq., 60% in mineral oil) in dry THF (50 mL) cooled to 0 ° C is added 6-amino-nicotinonitrile (3 , 4 g, 28.5 mmol, 1.1 eq.). After 30 min at 0 ° C, 2,6-dichloro-3-nitro-pyridine (5 g, 25.91 mmol, 1 eq.) Is added and the reaction is stirred at r.t. for 16 h. The reaction is cooled to 0 ° C, NaH (0.5 g, 13 mmol, 0.5 eq.) Is added and the reaction is stirred for 1 h at 0 ° C, then for 2 h at r.t. The reaction mixture is poured into ice water and stirred for 2 h. The precipitate is filtered, washed with H2O and air dried under vacuum. The solid obtained is captured in

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MeCN (75 mL), stirred at r.t. for 1 h 30 min and at 0 ° C for 1 h. Then, it is filtered and washed with MeOH to provide the desired compound.

[321] ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 10.81 (1H, br s), 8.73 (1H, dd), 8.61 (1H, d), 8.31 (1H, dd), 8.01 (1H, dd), 7.36 (1H, d).

1.2.2 General method B ^NO 2 ^NO 2

Λ X _____ _R2 Λ X

Cl N NH N N NH

R R1 R

[322] To the 6-chloro-3-nitro-pyridin-2-ylamino derivative (1 eq.) In DMSO is added the corresponding amine (1.1 eq.) And DIPEA (2 eq.), The reaction mixture it is then subjected to the microwave at 110-130 ° C until the end of the reaction. The mixture is diluted with Η ₂ Ο, the precipitate is filtered and air dried under vacuum to provide the desired compound.

Illustrative synthesis of general method B: 6- [3-nitro-6- (tetrahydro-pyran-4-ylamino) -pyridin-2-ylamino] nicotinonitrile (Int 8)

[323] To 6- (6-chloro-3-nitro-pyridin-2-ylamino) -nicotinonitrile (Int 1, 4 g, 14.51 mmol, 1 eq.) In DMSO (20 mL) is added tetrahydro-pyran -4-ylamine (1.65 mL, 15.96 mmol, 1.1 eq.) And DIPEA (5.05 mL, 29.02 mmol, 2 eq.), The reaction mixture is then subjected to the micro oven at 130 ° C for 20 min. The mixture is diluted with H2O and Et ₂ O, the precipitate is filtered and air dried under vacuum to provide the desired compound.

[324] ¹ H NMR (300 MHz, DMSO-d6) δ 11.38 (1H, s), 8.78 (1H, d), 8.47-8.63 (2H, m), 8.39 ( 1H, dd), 8.17 (1H, d), 6.27 (1H, d), 3.98-4.12 (1H, m), 3.91 (2H, d), 3.52 (2H , t), 1.94 (2H, d), 1.33-1.67 (2H, m).

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1.2.3 General method C

R1

GO

R1 R

[325] NBS (1.1 t to 2 eq.) Is added to a solution of the 3-nitro-pyridine-2,6-diamino derivative (1 eq.) In dry MeCN, the reaction is stirred at rt and monitored by UPLC-MS. If complete completion is not achieved, more NBS is added until there is no more starting material left. The precipitate formed is filtered, washed with Et ₂ O and air dried under vacuum to provide the desired compound.

Illustrative synthesis of the general method C: 6- [5-bromo-3-nitro-6- (tetrahydro-pyran-4-ylamino) -pyridin-2ylamino] -nicotinonitrile (Int 11)

AT THE;

CN

CN

[326] NBS (2.04 g, 11.46 mmol, 1.3 eq.) Is added to a solution of 6- [3-nitro-6- (tetrahydro-pyran-4-ylamino) -pyridin- 2-ylamino] nicotinonitrile (Int 8, 3 g, 8.81 mmol, 1 eq.) In dry MeCN (150 ml) and the reaction is stirred at rt for 4 h. NBS (0.31 g, 1.76 mmol, 0.2 eq.) Is added and the reaction is stirred at rt for an additional 16 h. The precipitate formed is filtered, washed with Et ₂ O and air dried under vacuum to provide the desired compound.

[327] ¹ H NMR (300 MHz, DMSO-d6) δ 11.10 (1H, br s), 8.80 (1H,

m), 8.34-8.50 (3H, m), 7.72 (1H, d), 4.05-4.25 (1H, m), 3.93 (2H, m), 3.38 -3.55 (2H, m), 1.70-1.87 (4H, m).

1.2.4 General method D

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R1

N NH R

R3

R2 -oL 'N ^ N ^ NH

I I

R1 R

[328] LiOtBu (3 eq.) Is added gradually to a solution of the corresponding alcohol (5 eq.) In dry 1,4-dioxane or to the corresponding alcohol used as the solvent. The 2-amino-3-bromo pyridine derivative (1 eq.) Is then added, followed by Cul (0.6 eq.). The reaction is heated to 80-120 ° C or 110-150 ° C under microwave irradiation, until the reaction is complete. The mixture is poured into ice water or a 1 N aqueous solution of HCI is added. The precipitate is filtered and air dried under vacuum. The residue is then purified by flash chromatography on silica gel to obtain the desired compound.

Illustrative synthesis of the general method D: 6- [5- [2- (2-hydroxy-ethoxy) -ethoxy] -3-nitro-6- (tetrahydro-pyran-4ylamino) -pyridin-2-ylamino] -nicotinonitrile (Int 19)

HN N

AT THE.

Br

NH

CN

CN

[329] LiOtBu (2.87 g, 35.8 mmol, 3 eq.) Is added gradually to a solution of 2- (2-hydroxy-ethoxy) -ethanol (5.7 mL, 59.7 mmol, 5 eq.) in dry 1,4-dioxane (50 ml). 6- [5-bromo-3-nitro-6- (tetrahydro-pyran-4ylamino) -pyridin-2-ylamino] -nicotinonitrile (Int 11.5.0 g, 11.9 mmol, 1 eq.) Is added, followed by Cul (1.36 g, 7.2 mmol, 0.6 eq.). The reaction is then heated to 120 ° C for 4 h. The mixture is cooled to 0 ° C, a 1 N aqueous solution of HCI (50 ml) is added and the resulting mixture is stirred at r.t. for 20 min. The precipitate is filtered and dried in vacuo. The residue is then purified by flash chromatography on silica gel, eluting in MeOH from 0 to 5% in DCM to provide the desired compound.

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[330] MW (calcd): 444.45; MW (obsd): 445.18 ES +.

[331] ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.42 (1H, s), 8.75-8.79 (1H, m), 8.47-8.50 (1H, m) , 8.38 (1H, dd), 7.87 (1H, d), 7.69 (1H, s), 4.58-4.72 (1H, m), 4.21 - 4.26 (2H , m), 4.11 - 4.21 (1H, m), 3.94 (2H, dd), 3.82 (2H, dd), 3.44-3.57 (6H, m), 1, 81-1.90 (2H, m), 1.74 (2H, qd).

1.2.5 General method E

YI ______ YI ^x >

^R2x NN ^X n ^R2x NN

R1 R R1 ^R

[332] To a solution of the 2,6-diamino-5-nitro-pyridine derivative (1 eq.) In dry MeOH are added trimethyl orthoformate (approximately 0.1 mL to 0.1 mmol of the 2.6 derivative) -diamino-5nitro-pyridine) and formic acid (approximately 0.1 mL for 0.1 mmol of 2,6-diamino-5-nitro-pyridine derivative). NH4CI (4 eq.) And Zn (4 to 5 eq.) Are then added and the mixture is heated to 70 ° C until completion of the reaction. The reaction mixture is then cooled to r.t.

[333] If, when cooling, precipitation is observed, the solid is filtered and subjected to aqueous processing with DCM / CHCl3 and a 2% aqueous formic acid solution to provide the desired compound.

[334] If, when cooling, no precipitation occurs, the solvents are evaporated and the residue is then purified by flash chromatography on silica gel to obtain the desired compound.

Illustrative synthesis of the general method E: 6- [6- [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3 -il] -nicotinonitrile (Compound 1)

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[335] To a solution of 6- [5- [2- (2-hydroxy-ethoxy) -ethoxy] -3-nitro-6 (tetrahydro-pyran-4-ylamino) -pyridin-2-ylamino] - nicotinonitrile (Int 19, 2.3 g, 5.2 mmol, 1 eq.) in dry MeOH (60 ml), trimethyl orthoformate (10 ml) and formic acid (10 ml) are added. NH4CI (1.1 g, 20.7 mmol, 4 eq.) And Zn (1.4 g, 20.7 mmol, 4 eq.) Are then added and the mixture is heated to reflux for 2 h. MeOH (30 ml) is added and the reaction mixture is heated to reflux for 1 h.

[336] The mixture is cooled to r.t., the precipitate formed is filtered and dried under vacuum. MeOH (100 ml) and formic acid (2 ml) are added and the resulting mixture is stirred under reflux for 1 h. The mixture is cooled to r.t., poured into ice water and the precipitate formed is filtered and dried under vacuum. The solid is suspended in a mixture of DCM and CHCl3, filtered through Celite and the filtrate is washed with a 2% aqueous formic acid solution. The organic phase is dried over Na2SO4, filtered and concentrated to dryness to provide the desired compound.

[337] MW (calcd): 424.46; MW (obsd): 425.40 ES +.

[338] ¹ H NMR (400 MHz, DMSO-d6) δ 9.00 (1H, dd), 8.89-8.95 (1H, m), 8.75 (1H, s), 8.62 ( 1H, dd), 7.59 (1H, s), 6.04 (1H, d), 4.61 - 4.69 (1H, m), 4.22 (2H, dd), 4.05-4 , 17 (1H, m), 3.90-3.98 (2H, m), 3.83 (2H, dd), 3.50-3.60 (6H, m), 1.99 (2H, m ), 1.55-1.70 (2H, m).

1.2.6 General method F

[339] A mixture of Compound 1, corresponding carboxylic acid (1.5 eq.), DMAP (1.5 eq.) And EDCI (2.25 eq.) Is stirred in DCM at r.t. until the reaction ends. The reaction is quenched with H2O, extracted with DCM and then the organic layer is dried over MgSCri and evaporated to dryness. The residue is purified by means of chromatography

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Illustrative summary of the general method F:

2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4-ylamino) 3H-imidazo [4,5-b] pyridin-6-yloxy] ester -ethoxy} (S) -2-tert-butoxycarbonylamino-3-methyl-butyric acid (Int 30)

HO,

CN

[340] A mixture of Compound 1 (42 mg, 0.1 mmol, 1 eq.), Boc- (S) -valine (33 mg, 0.15 mmol, 1.5 eq.), DMAP (19 mg, 0.15 mmol,

1.5 eq.) And EDCI (45 mg, 0.225 mmol, 2.25 eq.) Is stirred in DCM (5 ml) at r.t. for 3 h. The reaction is quenched with H2O, extracted with DCM, then the organic layer is dried over MgSCri and evaporated to dryness. The residue is purified by flash chromatography on silica gel, eluting with 0 to 100% EtOAc in heptanes to provide the desired compound.

1.2.7 General method G: Salification method

[341] The starting material is dissolved in hot EtOAc or a hot mixture of EtOAc and MeOH (5/1). Oxalic acid (0.2 M in EtOAc, 1 eq.) Is added to the hot solution. A precipitate forms, which is filtered, rinsed with Et2Ü and dried to provide the desired compound as the oxalic acid salt of the starting material.

Illustrative summary of the general method G:

2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4-ylamino) 3H-imidazo [4,5-b] pyridin-6-yloxy] ester -ethoxy} (S) -2amino-3-methyl-butyric acid, oxalic acid salt (Compound 10)

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[342] Compound 9 (100 mg, 0.19 mmol, 1 eq.) Is dissolved in hot EtOAc (10 mL) and oxalic acid (0.2 M in EtOAc, 0.96 mL, 0.19 mmol, 1 eq .) is added to the hot solution. The precipitate formed is filtered, rinsed with Et ₂ O and dried to provide the desired compound.

1.2.8 Synthesis of (3.4 cis) -3-Amino-tetrahydro-pyran-4-oi (Int 28)

1.2.8.1 Step i): 3,7-dioxa-bicycle [4,1,0] heptane

[343] To a solution of m-chloroperbenzoic acid (23.51 g,

136.2 mmol, 2 eq.) In DCM (15 mL) is added a solution of 3,6-dihydro-2H-pyrano (5.73 g, 68.1 mmol, 1 eq.) In DCM (10 mL) . The reaction mixture is allowed to stir at rt for 6 h, after which m-chloroperbenzoic acid (11.76 g, 68.1 mmol, 1 eq.) Is added. The reaction mixture is stirred at rt for 16 h and filtered. The filtrate is washed with saturated solutions of Na ₂ SOs, NaHCOs and water. The organic layer is then dried over Na ₂ SO4, filtered and concentrated in vacuo to provide the desired compound, used in the next step without further purification.

1.2.8.2 Step ii): (3.4 trans) -3-Benzylamino-tetrahydro-pyran-4-ol

[344] A mixture of 3,7-dioxa-bicyclo [4.1,0] heptane (2.7 mmol, 1 eq.) And benzylamine (300 pL, 2.7 mmol, 1 eq.) In EtOH (10 ml) is heated at reflux temperature for 18h. EtOH is then eva

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1.2.8.3 Step iii): N-Benzyl-N - ((3,4 trans) -4-hydroxy-tetrahydro-pyran-3-yl) -benzamide

[345] Benzoyl chloride (78 μΙ_, 0.68 mmol, 1 eq.) Is added dropwise to an ice-cold solution of (3.4 trans) -3-benzylamino-tetrahydro-pyran-4-ol from the previous step (140 mg, 0.68 mmol, 1 eq.) And TEA (280 μΙ_, 2.03 mmol, 3 eq.) In DCM (2 ml). The reaction mixture is stirred at rt for 1 h. The mixture is then washed twice with a 2 N aqueous solution of HCI. The aqueous layers are extracted with DCM and the combined organic layers are then dried over Na ₂ SO4, filtered and concentrated in vacuo to provide the desired compound.

1.2.8.4 Step iv): (3.4 cis) -3-Benzylamino-tetrahydro-pyran-4-ol

[346] A solution of N-benzyl-N - ((3.4 trans) -4-hydroxy-tetrahydro-pyran-3-yl) -benzamide (220 mg, 0.71 mmol, 1 eq.) In DCM ( 2.5 mL) is added dropwise to thionyl chloride (195 pL, 2.68 mmol,

3.8 eq.) At 0 ° C. The reaction mixture is stirred at rt for 4 h and then concentrated in vacuo. To the residue is added a 6 N aqueous solution of HCl (2 ml) and the resulting mixture is heated at reflux temperature for 18 h. After cooling, a precipitate is filtered, washed with water and the filtrate is extracted with EtOAc. To the aqueous layer is added Et ₂ O and a 2 N aqueous solution of NaOH is added to make the mixture alkaline. The phases are separated and the aqueous phase is extracted with DCM and EtOAc. The combined organic layers are then dried over Na ₂ SO4, filtered and concentrated in vacuo to provide the desired compound.

1.2.8.5 Step v): (3.4 cis) -3-Amino-tetrahydro-pyran-4-ol

[347] A solution of (3.4 cis) -3-benzylamino-tetra

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1.2.9 Synthesis of (cis-1,4) -1-methyl-4-methylamino-cyclohexanol (Int 29)

1.2.9.1 Step i): (cis-1,4) - (4-hydroxy-4-methyl-cyclohexyl) -carbamic acid tert-butyl ester

[348] To a suspension of cis-4-amino-1-methylcyclohexanol (1.0 g, 7.74 mmol, 1 eq.) In MeCN (15 mL) is added di-tert-butyl dicarbonate (1 , 85 g, 8.47 mmol, 1.1 eq.) And the mixture is stirred at rt for 16 h. The precipitate is filtered, washed with hexane and dried to provide the desired compound.

1.2.9.2 Step ii): (cis-1,4) -1-methyl-4-methylamino-cyclohexanol

[349] To a 2.0 M solution of L1AIH4 in THF (7 mL, 14.0 mmol, 4.9 eq.) Is added little by little tert-butyl acid ester (cis-1,4) - (4 -hydroxy-4-methyl-cyclohexyl) -carbamic acid (660 mg, 2.9 mmol, 1 eq.) in rt The reaction mixture is stirred at r.t. for 1 h and reflux for 45 min. The reaction mixture is cooled to r.t., then water and THF are added. The precipitate is filtered and washed with THF. The filtrate is concentrated to dryness, providing the desired compound.

1.2.10 intermediate 34: (2S) -2 - [[2- (tert-butoxycarbonylamino) acetyl] amino] -3-methyl-butanoate 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin6-yl] oxyethoxy] ethyl

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[350] Compound 9 (170 mg, 0.40 mmol, 1 eq.), 2- (tert-butoxycarbonylamino) acetic acid (Boc-Gly-OH, 105 mg, 0.60 mmol, 1.5 eq.), EDCI ( 173 mg, 0.90 mmol, 2.25 eq.) And DMAP (73 mg, 0.6 mmol,

1.5 eq.) Are mixed in DCM (4 ml) and stirred at r.t. overnight. The reaction is quenched with brine, extracted with DCM and the combined organic phases are evaporated to provide the desired compound.

Table II. Intermediates used for the compounds of the invention

SM = Initiation material, Mtd = Method, MS Mes’d = Measured mass

Int # Structures Name SM Mtd MW MS Mes'd 1 _ ^. NO ₂ jf I Cr'N '^ NH φ CN 6 - [(6-chloro-3-nitro-2- pyridyl) amino] pyridine-3-carbonitrile 2,6-dichloro- 3- nitropyridine THE 275.7 276.3 2 Z \ / ^NO2 Π HN ^ N ^ NH Ú 5 CN 6 - [[6 - [[(cis-3,4) -4-hydroxytetra- hydropyran-3-yl] amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 1 + Int 28 B 356.3 357.4 3 £ I HN ^ N ^ NH Ó ò O 0 Ón 6 - [[6 - [(1,1 -dioxotian-4-yl) amino] -3nitro-2-pyridyl] amino] pyridine-3carbonitrile Int 1 B 388.4 389.2 4 χΠ N ^ N ^ NH i i 'OH CN 6 - [[6 - [((cis-1,4) -4-hydroxy-4-methyl- cyclohexyl) -methyl-amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 1 + Int 29 B 382.4 -

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Int # Structures Name SM Mtd MW MS Mes'd 5 Z ^x v ' ^N ° ² Λ I HN ^ N ^ NH ô Φ' OH CN 6 - [[6 - [((cis1,4) -4-hydroxy-4-methyl- cyclohexyl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 1 B 368.4 - 6 χχ ^ ΝΟζ £ I HN ^ N ^ NH CN 6 - [[6 - [(3-hydroxycyclohexyl) amino] - 3-nitro-2-pyridyl] amino] pyridine-3- carbonitrile Int 1 B 354.4 355.4 7 _X - \ ^ ^NO 2 £ Y HN ^ N ^ NH powder ^F F CN 6 - [[6 - [(4,4-difluorocyclo- hexyl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 1 B 374.4 375.4 8 _χχ , ΝΟ ₂ IX HN ^ N ^ NH Ó ó CN 6 - [[3-nitro-6- (tetrahydropyran-4ylamino) -2-pyridyl] amino] pyridine-3carbonitrile Int 1 B 340.3 341.4 9 X ^ ^N ° 2 í X ^ N ^ N ^ NH Ó ó CN 6 - [[6- [methyl (tetrahydropyran-4- il) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 1 B 354.4 - 10 11 HN ^ N ^ NH ^H % A Jx _n V ψ CN 6 - [[5-bromo-6 - [[(cis-3,4) -4-hydroxytetrahydropyran-3-yl] amino] 3-nitro-2-pyridyl] amino] pyridine-3carbonitrile Int 2 Ç 435.2 435.4 437.3 11 ^ΒΓ Χ / ^ / ^Ν0 2 HN ^ N ^ NH Ô Ó CN 6 - [[5-bromo-3-nitro-6- (tetrahydropyran-4-ylamino) -2pyridyl] amino] pyridine-3-carbonitrile Int 8 Ç 419.2 419.5 421.4

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Int # Structures Name SM Mtd MW MS Mes'd 12 ^Br v ^/ ^ / ^N0 ' ¹ IX HN N NH 0 9 ^{0 0} CN 6 - [[5-bromo-6 - [(1,1-dixotian-4yl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 3 Ç 467.3 467.2 469.3 13 ^Br ^ X / ^N0 2 .11 N- ^ N ^ NH 9 9 'OH CN 6 - [[5-bromo-6 - [(4-hydroxy-4-methylcyclohexyl) -methyl-amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 4 Ç 461.3 - 14 Br II HN ^ N ^ NH 9 9 'OH CN 6 - [[5-bromo-6 - [((cis1,4) -4-hydroxy- 4-methyl-cyclohexyl) amino] -3-nitro-2- pyridyl] amino] pyridine-3-carbonitrile Int 5 ç 447.3 - 15 Br ^ / ^. NO ₂ 11 HN N NH Χοκφ CN 6 - [[5-bromo-6 - [(3-hydroxycyclohexyl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 6 ç 433.3 433.4 435.4 16 Br ^^ NOj IX HN ^ N ^ NH already ^F F CN 6 - [[5-bromo-6 - [(4,4-difluorocyclohexyl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 7 ç 453.3 453.3 455.3 17 Br ^^ NC ^ .XX N ^ N ^ NH ó 9 CN 6 - [[5-bromo-6- [methyl (tetrahydropyran-4-yl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 9 ç 433.3 433.4 435.4 18 ^ OH HN N NH ”Ú φ CN 6 - [[5- [2- (2-hydroxyethoxy) ethoxy] -6 [[(cis-3,4) -4-hydroxytetrahydropyran3-yl] amino] -3-nitro-2pyridyl] amino] pyridine-3 -carbonitrile Int 10 D 460.5 - 19 AT THE? XX HN ^ N ^ NH Ô i- ” CN 6 - [[5- [2- (2-hydroxyethoxy) ethoxy] -3nitro-6- (tetrahydropyran-4-ylamino) 2-pyridyl] amino] pyridine-3carbonitrile Int 11 D 444.5 445.2

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Int # Structures Name SM Mtd MW MS Mes'd 20 ^ΗΟ Χ ^ 0 ^ ⁰ γ ^ γ ^Ν ° 2 HN ^ N ^ NH Ó Í- ° ° CN 6 - [[6 - [(1,1 -dioxotian-4-yl) amino] -5- [2- (2-hydroxyethoxy) ethoxy] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 12 D 492.5 493.6 21 .0. ^ NO, <N N NH - ò Φ CN 6 - [[5- [2- (2-methoxyethoxy) ethoxy] -6 [methyl (tetrahydropyran-4-yl) amino] 3-nitro-2-pyridyl] amino] pyridine-3carbonitrile Int 17 D 472.5 473.6 22 ___ ° ^^ ^N ° 2 II HN ^ N ^ NH Ó Í- CN 6 - [[5- [2- (2-methoxyethoxy) ethoxy] -3nitro-6- (tetrahydropyran-4-ylamino) 2-pyridyl] amino] pyridine-3carbonitrile Int 11 D 458.5 459.5 23 ΗΟ ^ -ζχ ^ Ο, / χ, ΝΟ? 11 HN ^ N ^ NH ά _ΟΗ φ CN 6 - [[6 - [(3-hydroxy-cyclohexyl) amino] - 5- [2- (2-hydroxyethoxy) ethoxy] -3-nitro-2- pyridyl] amino] pyridine-3-carbonitrile Int 15 D 458.5 459.5 24 HO ^. / X ^ CI / xNQ, II HN ^ N ^ NH ô * ^F F CN 6 - [[6 - [(4,4-difluorocyclo- hexyl) amino] -5- [2- (2hydroxyethoxy) ethoxy] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 16 D 478.5 479.5 25 ° ^ ° τχ ^Ν ° ² r ^X N ^ N ^ NH - Φ Φ OH CN 6 - [[6 - [((cis1,4) -4-hydroxy-4-methyl- cyclohexyl) -methyl-amino] -5- [2- (2methoxyethoxy) ethoxy] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 13 D 500.6 - 26 , x \ .0. / ^ _, NO „CXY] HN N NH - i OH OH _CN 6 - [[6 - [((cis1,4) -4-hydroxy-4-methylcyclohexyl) amino] -5- [2- (2methoxyethoxy) ethoxy] -3-nitro-2pyridyl] amino] pyridine-3- carbonitrile Int 14 D 486.5 - 27 II HN ^ N NH Ó i 'OH CN 6 - [[5- [2- (2-hydroxyethoxy) ethoxy] -6- [((cis1,4) -4-hydroxy-4-methyl-cyclohexyl) amino] -3-nitro-2pyridyl] amino] pyridine-3-carbonitrile Int 14 D 472.5 - 28 nh ₂ nh ₂ HO ^ / L ^H0 - <® ⁺ ^ 0 (3,4-cis) -3-Amino-tetrahydro-pyran- 4-o I 3,6-di- hydro-2H- piran Example 0 117.2 -

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Int # Structures Name SM Mtd MW MS Mes'd 29 ^X NH '''OH (cis-1,4) -1-methyl-4-methylaminocyclohexanol cis-4amino-1methylcyclohexanol Example 0 143.2 - 30 Y 0 N 0 Xl?> Hn ^ n ^ -n ó Ü ^U CN (2S) -2- (tert-butoxycarbonylamino) - 3- [2- [2- [3- (5- cyano-2-pyridyl) -5- (tetrahydropyran- 4-ylamino) imidazo [4,5-b] pyridin-6- il] oxyethoxy] ethyl Cpd 1 F 623.7 624.9 31 0 N 0 r ° “4- ° n> HN ^ N ^ N 7 Q CN 2- (tert- butoxycarbonylamino) 2 [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl Cpd 1 F 581.6 582.7 32 0 ° * Ο ^Λ 0 JY XX ^x > HN ^ NN ώ Q CN 2- [tert- 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl butoxycarbonyl (methyl) amino] acetate Cpd 1 F 595.7 596.7 33 ° Y ^ -k. <° 0 ^ ° XX> HN ^ N I oh Q CN (2S) -pyrrolidine-1,2-dicarboxylate O2- [2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5-b] pyridin -6yl] oxyethoxy] ethyl] 1-tert-butyl Cpd 1 F 621.6 622.8 34 0 n à II ^H n ^ Yr XX ^x > HN ^ NN ô Y CN (2S) -2 - [[2- (tert- butoxycarbonylamino) acetyl] amino] - 3- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran- 4-ylamino) imidazo [4,5-b] pyridin-6- il] oxyethoxy] ethyl Cpd 9 Example 0 680.8 681.8 35 HN ^ O .0 <X AX <00 ^ ^ _n ^ ° WN XX '>HN' Ν '''N ô CN (2S) -2- (tert- butoxycarbonylamino) 01 - [2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl] 04 -tert-butyl Cpd 1 F 695.8 696.9

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Int # Structures Name SM Mtd MW MS Mes'd 36 HN ^ O r ° Ύ ô CN (2S) -2- (tert- butoxycarbonylamino) 01 - [2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl] 05 -erc-butyl Cpd 1 F 709.8 710.8 37 O JL yo ^ο ^ ^ο ϊΥ '> HN N ^N. ô ó CN 01 - [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl] 04-tert- butanedioate butyl Cpd 1 F 580.6 581.7

Example 2. Preparation of the compounds of the invention

2.1 Compound 3: 6- {6- [2- (2-hydroxy-ethoxy) -ethoxy] -5 - [((cis-1,4) -4-hydroxy-4-methyl-cyclohexyl) -methyl-amino] -imidazo [4,5-b] pyridin-3-yl} nicotinonitrile

[351] Formaldehyde (3.1 μΙ_, 0.11 mmol, 1 eq.) Is added to a solution of Compound 12 (50 mg, 0.11 mmol, 1 eq.) In a TFA / DCM mixture (2 mL , 1/1). After stirring at r.t. for 30 min, NaBH (OAc) 3 (47 mg, 0.22 mmol, 2 eq.) is added and the reaction is stirred for 1 h at r.t. The reaction mixture is then evaporated to dryness and the crude product is purified by means of preparative HPLC-MS to obtain the desired compound.

2.2 Compound 8: mono- (2- {2- [3- (5-cyano-pyridin-2-yl) -5 (tetrahydro-pyran-4-ylamino) -3H-imidazo [4,5-b ] pyridin-6-yloxy] ethoxy-ethyl) sulfuric acid

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[352] Compound 1 (84 mg, 0.2 mmol, 1 eq.) Is added to a solution of pyridine-SOs complex (127 mg, 0.8 mmol, 4 eq.) In pyridine (5 ml) and to reaction is heated to reflux for 16 h. The mixture is then evaporated to dryness and purified by flash chromatography on silica gel, eluting with 100% to 100% EtOAc (DCM / MeOH / AcOH / H ₂ O: 90/10/1/1 ), then 100% (DCM / MeOH / AcOH / H ₂ O: 85/15/2/2) to provide the desired compound.

2.3 Compound 9: 2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydropyran-4-ylamino) -3H-imidazo [4,5-b] pyridin-6 ester -yloxy] -ethoxy}-(S) -2-amino-3-methyl-butyric acid

[353] TFA (0.5 ml) is added to a solution of Int 30 (20 mg, 0.032 mmol, 1 eq.) In DCM (10 ml) and the mixture is stirred for 1 h at r.t. The reaction is quenched with the sat solution. aq. NaHCOs and extracted with EtOAc. The organic layer is dried over MgSO4 and evaporated to dryness. The residue is recrystallized from the DCM / Et2O solvent system in pentane to provide the desired compound.

2.4 Compound 16: 2- [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl 2-aminoacetate oxyethoxy] ethyl

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[354] Int 31 (195 mg, 0.34 mmol, 1 eq.) Is placed in a TFA / DCM mixture (1/5 ml) and the reaction is stirred at rt for 2 h. The reaction mixture is then diluted with toluene and evaporated to dryness. The residue is taken up in DCM, washed with a sat. aq.of NaHCOs and the organic phase is evaporated to dryness. The residue is dissolved in a minimal amount of DCM, a large volume of Et ₂ O is added and the solid formed is allowed to settle to the bottom of the flask. The solvents are carefully removed, leaving the solid in the flask, pentane is added and the solid is filtered to provide the desired compound.

2.5 Compound 17: 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6yl 2- (methylamino) acetate] oxyethoxy] ethyl

CN

[355] Int 32 (205 mg, 0.34 mmol, 1 eq.) Is placed in a TFA / DCM mixture (1/5 ml) and the reaction is stirred at rt for 2 h. The reaction mixture is then diluted with toluene and evaporated to dryness. The residue is taken up in DCM, washed with a sat. aq. NaHCOs and the organic phase is evaporated to dryness. The residue is dissolved in a minimal amount of DCM, a large volume of Et ₂ O is added and the solid formed is allowed to settle to the bottom of the flask. The solvents are carefully removed, leaving the solid in the flask, pentane is added and the solid is filtered to provide the desired compound.

2.6 Compound 18: 2- [2- [3- (5-cyano-) (2S) -pyrrolidine-2-carboxylate

2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6yl] oxyethoxy] ethyl

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[356] Int 33 (211 mg, 0.34 mmol, 1 eq.) Is placed in a TFA / DCM mixture (1/5 ml) and the reaction is stirred at rt for 2 h. The reaction mixture is then diluted with toluene and evaporated to dryness. The residue is taken up in DCM, washed with a sat. aq. NaHCOs and the organic phase is evaporated to dryness. The residue is dissolved in a minimal amount of DCM, a large volume of Et ₂ O is added and the solid formed is allowed to settle to the bottom of the flask. The solvents are carefully removed, leaving the solid in the flask, pentane is added and the solid is filtered to provide the desired compound.

2.7 Compound 19: 2- [2- [3- (5-cyano-2-pyridyl) -5- (2S) -2 - [(2-aminoacetyl) amino] -3-methyl-butanoate -5- (tetrahydropyran- 4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy] ethyl

[357] Int 34 (212 mg, 0.31 mmol, 1 eq.) Is placed in a TFA / DCM mixture (1/5 mL) and the reaction is stirred at r.t. for 2 h. Toluene is then added and the solvent is evaporated to dryness. The residue is dissolved in DCM and, after adding a sat. aq. From NaHCOs, a solid precipitates. It is filtered and dried to provide the desired compound.

2.8 Compound 27: (3S) -3-amino-4 [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo hydrochloric acid salt [4.5-

b] pyridin-6-yl] oxyethoxy] ethoxy] -4-oxo-butanoic

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OH

[358] A solution of HCI in 1,4-dioxane (4 M, 0.4 mL, 1.6 mmol, 5 eq.) Is added to Int 35 (222 mg, 0.32 mmol, 1 eq.) In

1,4-dioxane (4 ml). The mixture is stirred at r.t. for 3 h and then it is evaporated to dryness. The residue is taken up in a solution of HCI in 1,4-dioxane (4 M, 6 ml), the reaction is stirred at r.t. over 2 h and more solution of HCI in 1,4-dioxane (4 M, 2 ml) is added. After 1 h of stirring at r.t., the solvents are evaporated to dryness to provide the desired compound.

2.9 Compound 28: 4S acid hydrochloric acid salt) -4-amino-5 [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [ 4.5-

b] pyridin-6-yl] oxyethoxy] ethoxy] -5-oxo-pentanoic

[359] A solution of HCI in 1,4-dioxane (4 M, 0.41 mL, 1.63 mmol, 5 eq.) Is added to Int 36 (232 mg, 0.33 mmol, 1 eq.) In

1,4-dioxane (4 ml). The mixture is stirred at r.t. for 3 h and then it is evaporated to dryness. The residue is taken up in a solution of HCI in 1,4-dioxane (4 M, 6 ml) and the reaction is stirred at rt for 2 h. The solvents are evaporated to dryness to provide the desired compound.

2.10 Compound 33: 4- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5-b] pyridin-6-yl] oxyethoxy acid ] ethoxy] -4oxo-butanoic

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[360] Int 37 (90 mg, 0.16 mmol, 1 eq.) Is added to a TFA / DCM mixture (1/5 mL) and the reaction is stirred at r.t. for 2 h. Toluene is then added and the solvents are evaporated to dryness. The residue is purified by flash chromatography on silica gel, eluting with MeOH from 0 to 6% in DCM and the product obtained is precipitated from MeOH to provide, after filtration, the desired compound.

Table III. Illustrative compounds of the invention

SM = Initiation material, Mtd = Method, MS Mes’d = Measured mass

Cpd # Structure Name SM Method MW MS Mes'd 1 _O ^ OH --------- hn ^ n ^ n 0 i? CN 6- [6- [2- (2-hydroxy-ethoxy) ethoxy] -5- (tetrahydro-pyran4-ylamino) -imidazo [4,5b] pyridin-3-yl] nicotinonitrile Int 19 AND 424.5 ES + 425.4 2 o ^ ^OH HN NN Φ ó // \\ CN OO 6- (5- (1,1-dioxo-tetrahydro-2H-thiopyran-4ylamino) -6- [2- (2-hydroxyethoxy) -ethoxy] -imidazo [4,5b] pyridin-3-yl} nicotinonitrile Int 20 AND 472.5 ES + 473.6 ES- 471.5 3 .OH O NN ^N 0 Q Áqh CN 6- {6- [2- (2-hydroxy-ethoxy) ethoxy] -5 - [((cis-1,4) -4-hydroxy-4-methyl-cyclohexyl) -methyl-amino] imidazo [4,5-b ] pyridin-3-yl} nicotinonitrile Cpd 12 Example 0 466.5 ES + 467.6 4 THE \ N N N ô 5 CN 6- {6- [2- (2-methoxy-ethoxy) ethoxy] -5- [methyl- (tetrahydro-pyran-4-yl) -amino] imidazo [4,5-b] pyridin-3-yl} nicotinonitrile Int 21 AND 452.5 453.6

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Cpd # Structure Name SM Method MW MS Mes'd 5 _ <x The L X '> HN N N oh h CN 6- [6- [2- (2-methoxy-ethoxy) ethoxy] -5- (tetrahydro-pyran4-ylamino) -imidazo [4,5b] pyridin-3-yl] nicotinonitrile Int 22 AND 438.5 ES + 439.5 ES- 437.5 6 , OH 0 YX> HN N ^N. α CN 6- {5- (3-hydroxy-cyclo- hexylamino) -6- [2- (2hydroxy-ethoxy) -ethoxy] imidazo [4,5-b] pyridin-3-yl} nicotinonitrile Int 23 AND 438.5 ES + 439.49 7 Z \ ^ OH O Χλ ^χ > HN N ^N. ύ% FF ^CN 6- (5- (4,4-difluoro-cyclohexylamino) -6- [2- (2hydroxy-ethoxy) -ethoxy] imidazo [4,5-b] pyridin-3-yl} nicotinonitrile Int 24 AND 458.5 ES + 459.5 ES- 457.4 8 0 HOUUO S O k / ° v ^^ ^N ΤΎ '> HN N' N ô 0 ^u CN Mono- (2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4-ylamino) 3H-imidazo [4,5-b] pyridin6-yloxy] -etoxy} -ethyl) sulfuric acid Cpd 1 Example [351] 504.5 ES + 505.4 ES- 503.4 9 nh ₂ ° xX / χ - \ ^ Ο IO kz ° \ / ^ N Χχ '> HN N ^N , OQ ⁰ CN 2- {2- [3- (5-Cyanopyridin-2-yl) -5- (tetrahydropyran-4-ylamino) -3Himidazo [4,5-b] pyridin-6yloxy] -ethoxy} -ethyl acid ester (S) -2-amino-3methyl-butyric Int 30 Example 0 523.6 ES + 524.6 ES- 522.6 10 νη _ξ ° χΑ / x \ .OI 0 '' Χ / Οχ / ^^ Ν ΥΎ '> HN N ^N. 0 Q 0 ^CN JL ^ OH ^H0 X O 2- {2- [3- (5-Cyanopyridin-2-yl) -5- (tetrahydropyran-4-ylamino) -3Himidazo [4,5-b] pyridin-6yloxy] -ethoxy} -ethyl acid ester (S) -2-amino-3-methyl-butyric, oxalic acid salt Cpd 9 G 613.6 524.4

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Cpd # Structure Name SM Method MW MS Mes'd 11 _o -x / ° ^H XX '>hn' ^x n ⁿ . HO. Λ ' ^N. II \ 7 \ x ° CN 6- [6- [2- (2- hydroxyethoxy) ethoxy] -5 [[(cis-3,4) -4-hydroxytetrahydropyran-3yl] amino] imidazo [4,5b] pyridin-3-yl] pyridine-3carbonitrile Int 18 AND 440.5 441.3 12 O k / ° V% ^ N Y Ύ> HN N ^N , õ Q OH CN 6- [6- [2- (2- hydroxyethoxy) ethoxy] -5 [((cis-1,4) -4-hydroxy-4methylcyclohexyl) amino] imidazo [4,5b] pyridin-3-yl] pyridine-3carbonitrile Int 27 AND 452.5 453.0 13 O Υ ^ Ο'χΧ '^^ Ν xXI ^x> N ^ N ^N. OH CN 6- [5 - [((cis-1,4) -4-hydroxy- 4-methyl-cyclohexyl) -methylamino] -6- [2- (2methoxyethoxy) ethoxy] imidazo [4,5b] pyridin-3-yl] pyridine-3carbonitrile Int 25 AND 480.6 481.4 14 .THE. O ^ ✓θΧγ / ^ Ν YX ^x > HN N ^N. OH CN 6- [5 - [((cis-1,4) -4-hydroxy- 4-methylcyclohexyl) amino] - 6- [2- (2methoxyethoxy) ethoxy] imidazo [4,5b] pyridin-3-yl] pyridine-3carbonitrile Int 26 AND 466.5 467.3 15 Ck / \ / n í °% ^ x ^ ox ^^ m YX ^x > HN ^ N ^ N ô Ç ⁰ CN 2- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2- (dimethylamino) acetate] Cpd 1 F 509.6 510.4 16 O <x 1 ^NH 2 (° 0 <n ⁰ N XX ^x> HN ^ N ^ N Ò 0 ⁰ CN 2- [2 [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2-aminoacetate Int 31 Example 0 481.5 482.4

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Cpd # Structure Name SM Method MW MS Mes'd 17 í ° \ XXX hn ^ n ^ n ó i? ^u CN 2- (methylamino) acetate 2- [2- [3- (5-cyano-2-pyridyl) - 5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl Int 32 Example 0 495.5 496.3 18 ΗΝ'Λ C ° 0 A --- ⁰ V% U · xxx Htt ^N , 4 Q ^U CN 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl (2S) -pyrrolidine-2-carboxylate] Int 33 Example 0 521.6 522.5 19 o Λ ___ · ΝΗ XN ² c ° ^ _n ^ z ⁰ VVN XXX HN ^^ N ^N. O 6 CN (2S) -2 - [(2- amino [2- [2 [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl Int 34 Example 0 580.6 603.4 (M + Na) 20 I ^N | / ° k ^ O k ^ ⁰ V ^ N XXX> hn ^ n ^ n ó X CN 2-morpholinoacetate 2- [2- [3- (5-cyano-2-pyridyl) - 5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl Cpd 1 F 551.6 552.4 21 Οχ / \ i ^N ] / ° k ^ N ^ XXX hn ^ n ^ n ò x CN 2- [2- [3- [5 (cyano-2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2- (4-methylpiperazin-1yl) acetate] Cpd 1 F 564.6 565.3 22 1 C ^ Z'® ¹ · '. ⁰ D5 HN N ^N. ô ó CN 3- 2- [2- [3- (5-cyano-2- (dimethylamino) propanoate pyridyl) -5- (tetrahydropyran4-ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl Cpd 1 F 523.6 524.3

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Cpd # Structure Name SM Method MW MS Mes'd 23 0 ^{1 0} XX ^x> HN j c ά ^u CN 2- [2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4-ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2- (dimethylamino) acetate, salt oxalic acid Cpd 15 G 599.6 ES + 510.3 24 0 0. / χ JL .OH y NH ₂ HO Y -0 0 XX ^x > Ó 0 ^U CN 2- [2 [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2-aminoacetate, oxalic acid salt Cpd 16 G 571.6 ES + 482.3 25 ° γν. “V / ° 0 ^ _η ^ χ / Ο ^ χ_ _Ν XX ^x > ΗΝ ^ Ν ^ ~ Ν ó <? ^U CN 2- [2- [2- [3- (5-cyano-2-pyridyl) 5- (tetrahydropyran-4-amino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2- (methylamino) acetate, salt oxalic acid Cpd 17 G 585.6 ES + 496.3 26 ΗΝΛ 0 OJ ^ / X _OH. ^H0 T, 0 0 L .. ^ Cy ^ N XX ^x >ην'ν'' ^ Ó ó ^u CN 2- [2- [3- (5-cyano-2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl (2S) -pyrrolidine-2-carboxylate] ethyl, acid salt oxalic Cpd 18 G 611.6 ES + 522.3 27 nh ₂ 0 O.xÁ JL γ OH ó ₀ ⁰ TT '> hn ^ n- ^ n HCI Λ <h 0 ^ CN (3S) -3-Amino-4- [2 [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridine hydrochloric acid salt 6il] oxyethoxy] ethoxy] -4-oxobutanoic Int 35 Example 0 576.0 ES + 540.5

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Cpd # Structure Name SM Method MW MS Mes'd 28 νη _ξ , 0 0 --- °> x ^ Ss ^ N TT '> HN ^ N ^N. = 2nd Q \ CN (4S) -4-Amino-5- [2 [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridine hydrochloric acid salt 6il] oxyethoxy] ethoxy] -5-oxopentanoic Int 36 Example [358] 590.0 ES + 554.6 29 0 JL, NH, N ² XI ^x> 0 HN ^ N ^ ^N. -vo 3 CN 2- (2S) -2 - [(2aminoacetyl) amino] -3methyl-butanoate 2- [2 [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin- 6-yl] oxyethoxy] ethyl, oxalic acid salt Cpd 19 G 670.6 ES + 581.4 30 0. 1 ^N I XX '> 0 hn ^ n ^ n ”YÕ X CN 2 [2- [3- (5-cyano-2-pyridyl) -5 (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2-morpholinoacetate, oxalic acid salt Cpd 20 G 641.6 ES + 510.3 31 Ox / \ x \ 1 ^N 1 x ° Χ ^ χΝ ^ XX '>HN' NN õ 0 0 CN A .OH ^H0 T 0 2- [2- [3- [5 (cyano-2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl 2- (4-methylpiperazin-1yl) acetate, oxalic acid salt Cpd 21 G 654.6 ES + 565.6 32 Ο ^ χ- ^ χΧ H 1> X .OH l. ° t ( ₀ _ ⁰ <x ~ \ xO./5^ ^N XI ^x > hn ^ n ^ n Ó Ó CN 3- 2- [2- [3- (5-cyano-2- (dimethylamino) propanoate pyridyl) -5- (tetrahydropyran4-ylamino) imidazo [4,5b] pyridin-6-yl] oxyethoxy] ethyl, oxalic acid salt Cpd 22 G 613.6 ES + 524.4

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Cpd # Structure Name SM Method MW MS Mes'd 33 O y oh ° rx> HN N ^N. Ô Q CN 4- [2- [2- [3- (5-cyano- 2-pyridyl) -5- (tetrahydropyran-4ylamino) imidazo [4,5b] pyridin-6yl] oxyethoxy] ethoxy] -4-oxobutanoic Int 37 Example 0 524.5 ES + 525.3

Table IV. NMR data of representative compounds of the invention

Cpd # NMR data 1 ¹ H NMR (400 MHz, DMSO-de) δ 9.00 (1H, dd), 8.89-8.95 (1H, m), 8.75 (1H, s), 8.62 (1H, dd ), 7.59 (1H, s), 6.04 (1H, d), 4.61 - 4.69 (1H, m), 4.22 (2H, dd), 4.05-4.17 ( 1H, m), 3.90-3.98 (2H, m), 3.83 (2H, dd), 3.50-3.60 (6H, m), 1.98-2.01 (2H, m), 1.55-1.70 (2H, m) 2 ¹ H NMR (400 MHz, DMSO-de) δ 8.87-9.06 (2H, m), 8.63 (1H), 7.65 (1H, br s), 6.38 (1H, d) , 4.64 (1H, br s), 4.17-4.30 (3H, m), 3.80-3.88 (2H, m), 3.50-3.60 (4H, m), 3.40-3.48 (2H, m), 3.09-3.19 (3H, m), 2.15-2.35 (4H, m) 3 ¹ H NMR (400 MHz, DMSO-de) δ 8.98-9.03 (1H, m), 8.94 (1H), 8.87 (1H, s), 8.58 (1H, dd), 7.66 (1H, s), 4.58-4.70 (1H, m), 4.13-4.22 (2H, m), 4.08 (1H, s), 3.85-3, 96 (1H, m), 3.80 (2H, dd), 3.46-3.58 (4H, m), 2.93 (3H, s), 1.91-2.07 (2H, m) , 1.62 (2H, d), 1.50 (2H, d), 1.35-1.45 (2H, m), 1.13 (3H, s) 4 ¹ H NMR (300 MHz, CDCI ₃ ) δ 9.02-9.11 (1H, m), 8.93 (1H, s), 8.75 (1H, d), 8.13 (1H, dd) , 7.51 (1H, s), 4.16-4.29 (3H, m), 4.09 (2H, dd), 3.85-3.97 (2H, m), 3.67-3 , 74 (2H, m), 3.55-3.63 (2H, m), 3.41-3.55 (2H, m), 3.40 (3H, s), 3.00 (3H, s ), 2.01 (2H, qd), 1.76 (2H, dd) 5 ¹ H NMR (400 MHz, DMSO-de) δ 9.00 (1H, dd), 8.92 (1H, dd), 8.76 (1H, s), 8.62 (1H, dd), 7, 60 (1H, s), 6.00 (1H), 4.21 (2H, dd), 4.06-4.17 (1H, m), 3.90-3.97 (2H, m), 3 , 80-3.84 (2H, m), 3.61-3.66 (2H, m), 3.54 (2H, td), 3.47-3.51 (2H, m), 3.27 (3H, s), 1.99 (2H, dd), 1.54-1.67 (2H, m) 6 ¹ H NMR (400 MHz, DMSO-de) δ 8.99 (1H, d), 8.93 (1H, d), 8.74 (1H, s), 8.58 (1H, dd), 7, 55 (1H, s), 6.19 (1H, d), 4.72 (1H, d), 4.61 - 4.68 (1H, m), 4.15-4.23 (2H, m) , 3.89-4.02 (1H, m), 3.77-3.87 (2H, m), 3.67 (1H, m), 3.50-3.57 (4H, m), 2 , 17 (1H, d), 1.87-1.96 (1H, m), 1.71-1.86 (2H, m), 1.14-1.45 (4H, m) 7 ¹ H NMR (400 MHz, DMSO-de) δ 8.86-9.07 (2H, m), 8.76 (1H, br s), 8.63 (1H, d), 7.58 (1H, br s), 6.13 (1H, d), 4.51-4.76 (1H, m), 4.13-4.26 (2H, m), 3.72-3.94 (2H, m ), 3.47-3.63 (5H, m), 2.09 (8H, m) 8 ¹ H NMR (300 MHz, CD3OD) δ 9.03 (1H, d), 8.88-8.95 (1H, br s), 8.84 (1H, d), 8.41 (1H, dd) , 7.46 (1H, br s), 4.14-4.36 (5H, m), 3.95-4.12 (4H, m), 3.86 (2H, dd), 3.60- 3.76 (2H, m), 2.12 (2H, d), 1.64-1.85 (2H, m)

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Cpd # NMR data 9 ¹ H NMR (400 MHz, CDCl3) δ 9.02 (1H, d), 8.83 (1H, s), 8.76 (1H, brs), 8.13 (1H, d), 7.42 ( 1H, brs), 5.31 (1H, d), 4.36 (2H, brs), 4.24 (2H, brs), 4.18 (1H, brs), 4.08 (2H, d), 3.91 (2H, brs), 3.81 (2H, d), 3.62 (2H, t), 3.36 (1H, d), 2.16 (2H, d), 2.05 (1H , d), 1.26 (2H, br s), 0.98 (3H, d), 0.91 (3H, d) 10 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, dd), 8.93 (1H, dd), 8.78 (1H, s), 8.64 (1H, dd), 7, 60 (1H, s), 6.00 (1H, d), 4.40-4.46 (1H, m), 4.21-4.31 (3H, m), 4.09-4.18 ( 1H, m), 3.91-3.99 (2H, m), 3.85-3.90 (2H, m), 3.83 (1H, s), 3.75-3.80 (2H, m), 3.51-3.60 (2H, m), 2.04-2.15 (1H, m), 1.96-2.04 (2H, m), 1.58-1.68 ( 2H, m), 0.89-0.98 (6H, m) 11 ¹ H NMR (500 MHz, DMSO-de) δ 9.00 (1H, d), 8.84 (1H, d), 8.76 (1H, s), 8.59 (1H, dd), 7, 60 (1H, s), 5.80 (1H, d), 5.25 (1H, br s), 4.65 (1H, br s), 4.20-4.27 (2H, m), 4 , 16 (1H, tt), 4.02-4.10 (1H, m), 3.79-3.84 (2H, m), 3.66-3.75 (2H, m), 3.56 -3.62 (2H, m), 3.54 (4H, s), 1.82-1.94 (1H, m), 1.65-1.76 (1H, m) 12 ¹ H NMR (600 MHz, DMSO-de) δ 8.86-8.99 (2H, m), 8.69 (1H, s), 8.49-8.58 (1H, m), 7.50 (1H, s), 5.82 (1 H, d), 4.58-4.71 (1H, m), 4.17 (2H, d), 4.11 (1H, s), 3.81 (2H, d), 3.68-3.77 (1H, m), 3.53 (4H, s), 1.71-1.86 (4H, m), 1.62 (2H, d), 1.44-1.53 (2H, m), 1.17 (3H, s) 13 ¹ H NMR (500 MHz, DMSO-de) δ 9.00 (1H, d), 8.93 (1H, d), 8.87 (1H, s), 8.58 (1H, dd), 7, 65 (1H, s), 4.11 - 4.21 (2H, m), 4.08 (1H, s), 3.89 (1H, tt), 3.72-3.82 (2H, m) , 3.55-3.62 (2H, m), 3.44-3.50 (2H, m), 3.25 (3H, s), 2.93 (3H, s), 1.99 (2H , qd), 1.62 (2H, d), 1.50 (2H, d), 1.40 (2H, td), 1.12 (3H, s) 14 ¹ H NMR (300 MHz, DMSO-de) δ 8.92-8.99 (2H, m), 8.73 (1H, s), 8.59 (1H, dd), 7.54 (1H, s ), 5.79 (1H, d), 4.18 (2H, t), 4.08 (1H, br s), 3.80 (2H, t), 3.72-3.82 (1H, m ), 3.60-3.68 (2H, m), 3.45-3.52 (2H, m), 3.26 (3H, s), 1.72-1.83 (4H, m), 1.57-1.68 (2H, m), 1.43-1.56 (2H, m), 1.17 (3H, s) 15 ¹ H NMR (400 MHz, CDCI ₃ ) δ 9.03 (1H, dd), 8.83 (1H, s), 8.76 (1H, dd), 8.14 (1H, dd), 7.40 (1H, s), 5.42 (1H, d), 4.37- 4.39 (2H, m), 4.07-4.26 (5H, m), 3.93-3.95 (2H , m), 3.82-3.84 (2H, m), 3.61-3.67 (2H, m), 3.24 (2H, s), 2.37 (6H, s), 2, 14-2.24 (2H, m), 1.66-1.76 (2H, m) 16 ¹ H NMR (400 MHz, CDCl3) δ 9.05 (1H, d), 8.85 (1H, s), 8.79 (1H, d), 8.16 (1H, dd), 7.44 ( 1H, s), 5.34 (1H, d), 4.38-4.40 (2H, m), 4.08-4.28 (5H, m), 3.94-3.96 (2H, m), 3.83-3.85 (2H, m), 3.62-3.68 (2H, m), 3.52 (2H, s), 2.37 (6H, s), 2.15 -2.22 (2H, m), 1.65-1.75 (2H, m) 17 ¹ H NMR (400 MHz, CDCl 3) δ 9.05 (1H, dd), 8.85 (1H, s), 8.79 (1H, dd), 8.16 (1H, dd), 7.43 ( 1H, s), 5.36 (1H, d), 4.38- 4.41 (2H, m), 4.08-4.28 (5H, m), 3.94-3.96 (2H, m), 3.83-3.85 (2H, m), 3.62-3.68 (2H, m), 3.45 (2H, s), 2.48 (3H, s), 2.15 -2.22 (2H, m), 1.68-1.76 (2H, m) 18 ¹ H NMR (400 MHz, CDCl3) δ 9.05 (1H, d), 8.85 (1H, s), 8.79 (1H, d), 8.16 (1H, dd), 7.43 ( 1H, s), 5.38 (1H, d), 4.37-4.40 (2H, m), 4.08-4.27 (5H, m), 3.94-3.96 (2H, m), 3.82-3.86 (3H, m), 3.62-3.68 (2H, m), 3.08-3.14 (1H, m), 2.91-2.97 ( 1H, m), 2.12-2.21 (3H, m), 1.86-1.95 (1H, m), 1.60-1.82 (4H, m)

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Cpd # NMR data 19 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, d), 8.93 (1H, d), 8.77 (1H, d), 8.64 (1H, dd), 8, 30 (1H, br s), 7.61 (1H, s), 6.03 (1H, d), 5.14 (2H, br s), 4.09- 4.35 (6H, m), 3 , 91-3.98 (2H, m), 3.85-3.87 (2H, m), 3.74-3.76 (2H, m), 3.52-3.59 (2H, m) , 3.35 (2H, br s), 1.96-2.10 (3H, m), 1.58-1.68 (2H, m), 0.85-0.88 (6H, m) 20 ¹ H NMR (400 MHz, CDCl3) δ 9.03 (1H, d), 8.83 (1H, s), 8.77 (1H, d), 8.14 (1H, dd), 7.42 ( 1H, s), 5.34 (1H, d), 4.36- 4.39 (2H, m), 4.07-4.26 (5H, m), 3.93-3.95 (2H, m), 3.81-3.84 (2H, m), 3.74-3.77 (4H, m), 3.61-3.67 (2H, m), 3.28 (2H, s) , 2.59-2.61 (4H, m), 2.16-2.20 (2H, m), 1.65-1.75 (2H, m) 22 ¹ H NMR (400 MHz, CDCl3) δ 9.04 (1H, dd), 8.84 (1H, s), 8.77 (1H, dd), 8.14 (1H, dd), 7.42 ( 1H, s), 5.35 (1H, d), 4.33- 4.36 (2H, m), 4.07-4.27 (5H, m), 3.94-3.96 (2H, m), 3.81-3.83 (2H, m), 3.61-3.68 (4H, m), 2.54-2.68 (4H, m), 2.27 (6H, s) , 2.16-2.20 (2H, m), 1.66-1.76 (2H, m) 23 ¹ H NMR (400 MHz, DMSO-de) δ 9.00 (1H, dd), 8.92 (1H, dd), 8.77 (1H, s), 8.63 (1H, dd), 7, 60 (1H, s), 6.02 (1H, d), 4.21 - 4.27 (4H, m), 4.06 - 4.18 (1H, m), 3.91 - 3.95 ( 2H, m), 3.84-3.86 (2H, m), 3.73-3.76 (2H, m), 3.62-3.68 (2H, brs), 3.51-3, 57 (2H, m), 2.5 (6H, s), 1.96-2.01 (2H, m), 1.57-1.67 (2H, m) 24 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, dd), 8.93 (1H, dd), 8.78 (1H, s), 8.64 (1H, dd), 7, 62 (1H, s), 6.04 (1H, d), 4.32-4.35 (2H, m), 4.24-4.26 (2H, m), 4.09-4.18 ( 1H, m), 3.92-3.97 (2H, m), 3.86-3.89 (2H, m), 3.84 (2H, s), 3.77-3.79 (2H, m), 3.53-3.59 (2H, m), 1.98-2.02 (2H, m), 1.58-1.68 (2H, m) 25 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, dd), 8.93 (1H, dd), 8.78 (1H, s), 8.64 (1H, dd), 7, 62 (1H, s), 6.04 (1H, d), 4.33-4.35 (2H, m), 4.23-4.26 (2H, m), 4.09-4.18 ( 1H, m), 3.92-3.98 (4H, m), 3.86-3.89 (2H, m), 3.76-3.79 (2H, m), 3.52-3, 59 (2H, m), 2.58 (3H, s), 1.96-2.04 (2H, m), 1.58-1.68 (2H, m) 26 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, dd), 8.93 (1H, dd), 8.78 (1H, s), 8.64 (1H, dd), 7, 62 (1H, s), 6.04 (1H, d), 4.29-4.42 (3H, m), 4.23-4.26 (2H, m), 4.10-4.18 ( 1H, m), 3.92-3.97 (2H, m), 3.87-3.89 (2H, m), 3.77-3.80 (2H, m), 3.53-3, 59 (2H, m), 3.12-3.25 (4H, m), 2.19-2.27 (1H, m), 1.92-2.03 (3H, m), 1.81 1.89 (2H, m), 1.58-1.68 (2H, m) 28 ¹ H NMR (400 MHz, DMSO-de) δ 9.04 (1H, dd), 8.92-8.94 (H, m), 8.78 (1H, s), 8.66 (1H, dd ), 8.54 (3H, br s), 7.59 (1H, s), 4.38-4.43 (1H, m), 4.24-4.34 (3H, m), 4.08 -4.17 (2H, m), 3.87-3.97 (4H, m), 3.77-3.80 (2H, m), 3.51-3.56 (2H, m), 2 , 37-2.54 (2H, m), 1.97-2.06 (4H, m), 1.59-1.69 (2H, m) 30 ¹ H NMR (400 MHz, DMSO-de) δ 9.01 (1H, dd), 8.93 (1H, d), 8.77 (1H, s), 8.63 (1H, dd), 7, 61 (1H, s), 6.03 (1H, d), 4.21 - 4.27 (4H, m), 4.08 - 4.18 (1H, m), 3.91 - 3.98 ( 2H, m), 3.84-3.88 (2H, m), 3.72-3.76 (2H, m), 3.52-3.60 (6H, m), 3.32 (2H, s), 2.53-2.57 (4H, m), 1.96-2.04 (2H, m), 1.58-1.68 (2H, m) 31 ¹ H NMR (400 MHz, DMSO-de) δ 9.02 (1H, dd), 8.93 (1H, d), 8.78 (1H, s), 8.64 (1H, dd), 7, 62 (1H, s), 6.03 (1H, d), 4.08-4.26 (5H, m), 3.92-3.98 (2H, m), 3.84-3.88 ( 2H, m), 3.72-3.76 (2H, m), 3.52-3.59 (2H, m), 3.36 (2H, s), 2.60-3.15 (11H, m), 1.96-2.04 (2H, m), 1.58-1.68 (2H, m)

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Cpd # NMR data 32 ¹ H NMR (400 MHz, DMSO-de) δ 9.01 (1H, dd), 8.93 (1H, dd), 8.78 (1H, s), 8.63 (1H, dd), 7, 62 (1H, s), 6.04 (1H, d), 4.21 - 4.27 (4H, m), 4.09 - 4.18 (1H, m), 3.91 - 3.98 ( 2H, m), 3.84-3.89 (2H, m), 3.73-3.78 (2H, m), 3.52-3.59 (2H, m), 3.24-3, 28 (2H, t), 2.82-2.86 (2H, t), 2.73 (6H, s), 1.96-2.04 (2H, m), 1.57-1.67 ( 2H, m) 33 ¹ H NMR (400 MHz, DMSO-de) δ 12.19 (1H, br s), 9.02 (1H, dd), 8.94 (1H, dd), 8.77 (1H, s), 8 , 64 (1H, dd), 7.61 (1H, s), 6.03 (1H, d), 4.08-4.25 (5H, m), 3.91-3.97 (2H, m ), 3.84-3.88 (2H, m), 3.70-3.75 (2H, m), 3.51-3.59 (2H, m), 2.45-2.55 (4H , m), 1.96-2.03 (2H, m), 1.58-1.68 (2H, m)

BIOLOGICAL EXAMPLES

Example 3. In Vitro Tests

3.1. Determination of IC50 for human IRAK-4

[361] The IC50 value for IRAK-4 is determined in a radioactive filter plate assay. The assay principle consists in measuring the incorporation of ³³ P into the substrate RIP140 when phosphorylation by the enzyme IRAK-4 using [γ- ³³ Ρ] ΑΤΡ and ATP. Incorporated ^33P is not removed when loading the samples on a filter plate (using a collector, Perkin Elmer) and 6 subsequent washing steps. The ³³ P incorporated in RIP140 is measured by a scintillation counter (Topcount, PerkinElmer) after adding Microscint ™ -20 (PerkinElmer, 6013621) to the filter plates.

[362] 5 μΙ of a series of dilutions in water of the test compound (starting from 20 μΜ or a higher concentration of 6.6 μΜ) of a 100% DMSO stock solution, 1/5 dilution, are added to the wells (final DMSO concentration of 1% in the reaction assay). IRAK-4 (Carna Biosciences, 09-145) and RIP140 (SEQ ID1, cf. Table VII) are used at a final concentration of 10 ng / mL and 4 μΜ, respectively. The enzyme and substrate are diluted in 25 mM Tris, pH 7.5, 0.025% Triton X-100, 5 mM MnCh and 2 mM DTT for a total volume of 11 pL. The reaction is initiated by adding 9 pL of ATP to 1 μΜ (Sigma, A6419-5G) + 0.25 pCi of [γ- ³³ Ρ] ΑΤΡ (PerkinElmer, NEG602K001 MC), diluted in the same buffer as the enzyme and the substrate. The mixture is incubated at 30 ° C

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99/154 for 45 min. The reaction is terminated by adding 25 μΙ_ of 150 mM phosphoric acid (VWR, 1.00573.1000). The samples are transferred to filter plates and the built-in radioactivity is measured using a scintillation counter.

[363] Staurosporine at 10 μΜ (1% DMSO) is used as a positive control (100% inhibition); vehicle (water + 1% DMSO) as a negative control (0% inhibition).

Table V. IC50 for in vitro human IRAK-4 of the compounds of the invention

Cpd # IC ₅ o hlRAK-4 (nM) 1 6.35 2 25.4 3 21.7 4 523 5 13.8 6 4.85 7 14.3 8 51.4 9 13.5 11 14.3 12 0.95 13 39.1 14 3.45

3.2. Characterization of kinase selectivity profile (wide panel)

[364] Inhibition of human kinases is determined in radiometric kinase assays at REACTION BIOLOGY (Reaction Biology Corp., 1 Great Valley Parkway, Suite 2 Malvern, PA 19355, USA).

[365] To determine its IC50, a compound is tested in 10 doses from 10 μΜ (highest concentration), with dilutions in

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100/154 series of 3 times. IC50 values are derived by adjusting the dose-response curves to the% Enzyme Activity Remaining (in relation to the DMSO controls).

3.3. Characterization of kinase selectivity profile (focused panel)

[366] The purpose of this trial is to determine the activity and selectivity of a compound of the invention in a selected range of human kinases that can result in undesirable side effects when inhibited (Dy and Adjei, 2013; Force and Kolaja, 2011).

3.3.1. Test protocol

[367] The IC50 value for off-target kinases is determined in radioactive filter plate assays. The assay principle consists of measuring the incorporation of ³³ P into a peptide substrate when phosphorylating by the enzyme kinase using [γ- ³³ Ρ] ΑΤΡ and ATP. Incorporated ^33P is not removed when loading the samples on a filter plate (using a collector, Perkin Elmer) and 6 subsequent washing steps. The ³³ P incorporated in the peptide substrate is measured by a scintillation counter (Topcount, PerkinElmer) after adding Microscint ™ -20 (PerkinElmer, 6013621) to the filter plates.

[368] 5 μΙ of a series of dilutions in water of the test compound (from 20 μΜ or a higher concentration of 6.6 μΜ) of a mother solution in 100% DMSO, dilution of 1/5, are added to the wells (final DMSO concentration of 1% in the reaction assay). Enzyme and peptide substrate are used in optimized concentrations (see Table VI). The enzyme and the substrate are diluted in assay buffer to a total volume of 11 μΙ_. The reaction is initiated by adding 9 μΙ_ of ATP + [γ- ³³ Ρ] ΑΤΡ, diluted in the same buffer as the enzyme and 0 substrate. The mixture is incubated at 30 ° C. The reaction is terminated by adding 25 μΙ_ of 150 mM phosphoric acid. The samples are transferred to filter plates and the radioacti

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101/154 incorporated is measured using a scintillation counter.

[369] The incubation time, the composition of the assay buffer and the concentrations of ATP, enzyme and substrate are shown in Table VI, for example, off-target kinase assays.

[370] Staurosporine at 10 μΜ (1% DMSO) is used as a positive control (100% inhibition); vehicle (water + 1% DMSO) as a negative control (0% inhibition).

Table VI. Conditions for Forced-Target Kinase Inhibition Assays

Kinase, [Kinase] Substrate, [Substrate] ATP Test Buffer Incubation time GLA (Life Technologies, P3049), 40 ng / mL PolyGT (Sigma- Aldrich, P0275), 5 pg / mL 0.5 pM ATP + 0.25 pCi / 25 pL [y- ³³ P] ATP 50 mM Tris, pH of 7.7 0.03 Triton X-100 % 1 mM DTT 25 mM MgCÍ2a 60 min Aurora B (Carna BioSciences, 05-102), 10 ng / mL Histone H3 peptide (SEQ ID2), 0.5 pM 1.3 pM ATP + 0.25 pCi / 25 pL of [y- ³³ P] ATP 25 mM Tris, pH of 7.7 0.01 Triton X-100 % 5 mM MgCÍ2a 5 mM DTT 90 min CDK2 (Carna Biosciences, 04-103), 30 ng / mL Histone-derived peptide H1 (SEQ ID3), 0.36 pM 0.1 pM ATP + 0.25 pCi / 25 pL of [y- ³³ P] ATP 8 mM MOPS, pH 7.0 0.01% Brij-35 1 mM DTT MnCÍ2a 5 mM 60 min CDK9 (Millipore, 14-685), 230 ng / mL PDKtoid (SEQ ID4), 0.5 pM 0.25 pM ATP + 0.125 pCi / 25 pL [y- ³³ P] ATP 20 mM MOPS, pH 7.0 0.01 Triton X-100 % MnCÍ2a 5 mM 60 min c-KIT (Millipore, 14-559), 0.01 mU / pL PolyGT (Sigma- Aldrich, P0275), 0.1 mg / mL 3 pM ATP -ιΟ, 25 pCi / 25 pL of [y- ³³ P] ATP 16 mM Tris, pH of 7.0 500 pM EDTA 0.01 Triton X-100 % MnCÍ2a 10 mM 90 min

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Kinase, [Kinase] Substrate, [Substrate] ATP Test Buffer Incubation time 1 mM DTT 10 mM MgOAc GSK3b (Carna Biosciences, 04-141), 20 ng / mL Phospho glycogen synthase peptide 2 (Millipore, 12-241), 1.25 μΜ ATP at 1.5 μΜ + 0.25 pCi / 25 pL of [y- ³³ P] ATP 50 mM Tris, pH of 8.0 0.01% Brij-35 1 mM DTT 5 mM MgOAc 90 min c-SRC (Carna Biosciences, 08-173), 8 ng / mL PolyGT (Sigma- Aldrich, P0275), 2 pg / mL 0.25 pM ATP + 0.125 pCi / 25 pL [y- ³³ P] ATP 25 mM MOPS, pH 7.0 MnC ^ at 10 mM DTT at 2.5 mM Brij35 at 0.01% 60 min

Table VII. Peptide substrates used in off-target human kinase inhibition assays

Substrate SEQ ID AT THE. Sequence Provider Peptide RIP140 1 CYG VASS H LKTLLKKS KVKDQ Almac Group Ltda. 20 Seagoe Industrial Estate Craigavon BT63 5QD United Kingdom H3 histone peptide 2 ARTKQTARKSTGGKAPRKQLC AnaSpec Inc. 34801 Campus Drive Fremont, CA 94555 USA H1 histone-derived peptide 3 GGGPATPKKAKKL AnaSpec Inc. 34801 Campus Drive Fremont, CA 94555 USA PDKtoid 4 KTFCGTPEYLAPEVRREPRILSEEEQEMFRDFDYIADWC Thermo Fisher Scientific Inc. 81 Wyman St. Waltham, MA 02451 USA

Table VIII. IC50 of human kinase off-target in vitro with posts illustrating the invention

Cpd # GLA ICso (nM) Aurora B ICso (nM) CDK2 ICso (nM) CDK9 ICso (nM) GSK3b ICso (nM) c-SRC IC ₅₀ (nM) 1 - - > 6670 > 1330 > 6670 > 1330 2 - - > 1330 712 > 1330 > 1330 5 2690 - > 4000 2290 > 4000 > 2940

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Cpd # GLA IC ₅₀ (nM) Aurora B IC50 (nM) CDK2 IC50 (nM) CDK9 IC50 (nM) GSK3b IC50 (nM) c-SRC IC50 (nM) 6 656 - 2670 1030 > 4000 754 9 633 - > 4000 639 3250 286 11 - - > 1330 834 > 1330 > 1330 12 138 > 1880 944 1150 > 1330 477 14 - - 1380 1390 2240 319

3.3.2. Conclusion

[371] The data in Table VIII, relative to those in Table V, show the lowest inhibitory potency of the compounds of the invention of off-target kinases versus IRAK-4. These data confirm the selectivity of the compounds of the invention in relation to IRAK-4, thus limiting the risk of side effects associated with the inhibition of off-target kinases.

3.4. Cellular assay: inhibition of the release of TN0 α activated by CL097 in PBMCs

[372] The compounds of the invention are tested in a cellular assay using primary isolated human peripheral blood mononuclear cells (PBMCs) to measure the secretion of the inflammatory cytokine TNFα upon activation of TLR using the specific TLR7 / 8 agonist, CL097. The release of TNFα protein in the cell culture supernatant is quantified by an enzyme-linked immunosorbent assay protocol (ELISA) of human TNFα.

3.4.1. Isolation of primary human PBMCs from the human buffy coat

[373] A human buffy coat (provided by the Croatian Institute for Transfusion Medicine) is kept overnight at 4 ° C and processed the next day for isolation of PBMCs. PBMCs are isolated by means of density gradient centrifugation using Ficoll-Paque ™ PLUS (GE HealthCare, 171440-02). Equal volumes of a buffy coat are diluted 1: 4 with sterile PBS (1X) and 35 mL are carefully placed

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104/154 layered over 15 mL of Ficoll-Paque PLUS in appropriate 50 mL Falcon® tubes. The tubes are centrifuged for 35 min at 1500 rpm in rt without acceleration or pause. After centrifugation, the upper plasma layer is removed and the ring of mononuclear cells is carefully isolated and transferred to a new Falcon® tube. The isolated cell suspension is diluted in PBS to 50 mL, followed by a centrifugation step at 1300 rpm for 10 min in rt. After 2 additional steps of washing in PBS and cell separation, the remaining erythrocytes are subjected to lysis by resuspending the cell pellet in 50 ml of AKL lysis buffer (NH ₄ CI at 150 mM, NaHCO ₃ at 10 mM, In ₂ mTA EDTA, pH 7.4) followed by gentle mixing. The 50 mL suspension is then centrifuged at 1300 rpm for 10 min in rt, followed by removal of the supernatant and resuspension of the cell pellet in culture medium (RPMI 1640 (Gibco, 21875) + 10% fetal bovine serum (FBS , Biowest, S1810) thermally inactivated for 30 min at 56 ° C + Pen / Strep (Gibco, 15240)).

3..4.2. Compound treatment and activation in the PBMC assay

[374] Cells are counted using a hematology analyzer (Sysmex XS-500I) and placed at a density of 4.0 χ 10 ⁵ cells per well in 160 gL of culture medium in 96 well culture plates. Subsequently, PBMCs are pre-incubated with the test compound by adding 20 µl of 10X concentrated solution for 1 hour at 37 ° C and 5% CO2. The compounds are tested in different concentrations and prepared by 3-fold serial dilutions from the 10 mM stock solution in DMSO, followed by a 1:50 dilution step in 2X M199 medium (Gibco, 21157-029) supplemented with 1% FBS and 1% Pen / Strep. The final test concentrations in the assay start from 20 μΜ, with subsequent 3-fold serial dilutions and concentrate

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105/154 equal final sections of 0.2% DMSO. After the pre-incubation step of the compound, PBMCs are activated by adding 20 μΙ_ of a 10 pg / mL solution of CL097 (InvivoGen, tlrl-c97-5) to the wells with a final test volume of 200 μΙ_ per well and a final concentration of 1 pg / mL of CL097. Negative controls are adjusted to equal DMSO concentrations without the CL097 activator. The assay plates are then incubated for 4 h in a humidified incubator at 37 ° C and 5% CO2. The cell supernatants are then collected by transferring the cell medium to a plate with 384 deep wells and immediately transferred to the ELISA plate for quantification of human TNFα.

3.4.3. Quantification of TNFa by ELISA

[375] The levels of TNFα secreted in cell supernatants are quantified in an antibody capture activity assay (ELISA). A 384-well Greiner Lumitrac ™ plate is coated with 40 pL per well of a solution of 1 pg / ml of human anti-TNFα antibody solution (MAb1; BD Biosciences, 551220) diluted in PBS during a one-day incubation for The other at 4 ° C. After washing the wells with 100 µl of PBS, the remaining binding sites are blocked with 100 µl of blocking buffer (PBS + 1% bovine serum albumin + 5% sucrose) and incubated for 4 h at r.t. After the blocking step, the wells are washed once with PBS with Tween 20 (PBST), followed by the addition of samples and standards. Samples containing TNFα are diluted 1/3 in dilution buffer and 40 µl are added for overnight incubation at 0 ° C. The wells are then washed 3 times, twice with PBST and once with PBS, after the addition of 35 μl of secondary biotinylated anti-TNFα detection antibody (MAb11; BD Biosciences, 554511) in a diluted 1 / 2000 at the final concentration of 250 ng / mL. After 2 hours of incubation in r.t. and steps

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106/154 wash wells (2x PBST, 1x PBS), the wells are incubated with 35 μΙ_ of a solution of streptavidin conjugated to horseradish peroxidase diluted to 1/4000 (Life Technologies, SNN2004), followed by an incubation step of 45 min in rt in the dark. The wells are then washed 3 times (2x PBST, 1x PBS), followed by 5 min incubation with 50 µl of Chemiluminescence ELISA Substrate solution (Roche, 11582950001). The luminescent signal of the converted substrate is measured in a plate reader Multilabel PerkinElmer EnVision 2104.

3.4.4. Data analysis

[376] All controls are measured within the linear range of the ELISA human TNFα standard curve. All data are checked for validity in relation to the assay quality parameters (signal / background> 2 and Z '> 0.3).

[377] Non-activated samples (without activator / vehicle (0.2% DMSO)) are used as a positive control (100% inhibition). As a negative control (0% inhibition), activated samples are used (activator / vehicle (0.2% DMSO)). The positive and negative controls are used to calculate the values of Z 'and the percentage of inhibition (PIN), according to the following formula:

RCLUactivator / vehicle - RCLUtest compound

PIN = ----------; -; --—-------------------- ₁ , X 100

RCLlIctivator / vehicle - RCLUse activator / vehicle with RCLU = Relative Chemiluminescence Light Units.

[378] PIN values are plotted for compounds tested in concentration-response mode and IC50 values are derived using the GraphPad Prism® software, applying a non-linear (sigmoidal) regression curve fit.

3.5. Cellular assay: cancer cell assays

3.5.1. Cell lines

[379] Human lymphoma cells from OCI cell lines

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Ly3, OCI-Ly10, OCI-LY7 and OCI-Ly19 (from DSMZ, Germany or ATTC, USA) are grown in IMDM (Gibco®, 21980-032) supplemented with 10% fetal bovine serum (Invitrogen, S7524) or serum 20% human (Invitrogen, 34005100) at 37 ° C in 5% CO2.

3.5.2. Cell growth assay

[380] Lymphoma cells (2-7 χ 10 ³ ) are plated in 96-well plates and treated with different doses of test compounds from 30 μΜ (1/3, 8 point dilutions). The treated cells are incubated for 7 days at 37 ° C in 5% CO2. Staurosporine (10 μΜ) is used as a positive control.

[381] Cell growth is determined by incubating cells with alamarBIue® (Invitrogen, DAL 1025), according to the manufacturer's instructions. Fluorescence is measured using a PerkinElmer EnVision® plate reader. The percentage of growth inhibition is calculated using values for the DMSO vehicle as 0% inhibition values and values for staurosporine as 100% inhibition.

3.5.3. Cellular assay of response to IL-1 and TNF-α in SW1353 cells

[382] The purpose of this assay is to assess the selectivity of the compounds of the invention by the TLR / IRAK-4 pathway activated in a human cell assay scenario in vitro. SW1353 cells come from a chondrocyte cell line and are responsive to both the cytokine activator interleukin 1 (IL-1) and the activator TNFa. Both cytokine activators induce the expression of interleukin 6 (IL-6) and MMP13 by these cells. The IL-6 and MMP13 releases are used as readings in this assay and represent a measure for the level of inhibition of the TLR / IRAK-4 pathway by the tested compound. The IL-1 activator signals via an IRAK-4 dependent pathway, while 0 TNFa does not require IRAK-4 to signal

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108/154 tion. Therefore, compounds that selectively inhibit IRAK-4 have an impact only on IL-1 controlled expression of MMP13 or IL-6 by SW1353 cells and have no impact on TNFα controlled expression of these proteins.

3.5.4. SW1353 cell collection and culture

[383] SW1353 cells are cultured in DMEM supplemented with 10% FBS and 1% Penicillin / Streptomycin. The cells are incubated at 37 ° C in a humidified atmosphere of 5% CO ₂ and subcultured twice a week. During subculture, trypsin EDTA is used to separate cells, followed by a neutralization step with cell culture medium. After centrifugation (1000 rpm for 5 min), the pellet is resuspended in cell culture medium and cells are counted using an automatic cell counter (Invitrogen Countess ™).

[384] The cells are used in the 16-pass and placed at a density of 15,000 cells per well in 120 μΙ_ of cell culture medium in 96-well culture plates. The cells are allowed to set during overnight incubation.

3.5.5. Compound treatment and activation in the SW1353 assay

[385] SW1353 cells are pre-incubated with the test compound by adding 15 µl of 10x concentrated solution for 2 h at 37 ° C and 5% CO2. The compounds are tested in different concentrations and prepared by means of 3-fold serial dilutions of 10 mM stock solution in DMSO, followed by a 1/50 dilution step in cell culture medium. The final test concentrations in the assay start from 20 μΜ, with subsequent 3-fold serial dilutions with equal final concentrations of 0.2% DMSO. After the compound pre-incubation step, SW1353 cells are activated by adding 15 pL of concentrated IL-1 β 10X activator (Peprotech, 200-01B) or TNFa (Peprotech, 300-01 A)

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109/154 to wells with a final assay volume of 150 μΙ_ per well and a final activator concentration of 1 ng / mL and 10 ng / mL, respectively. Negative controls are adjusted with equal concentrations of DMSO without activator. The assay plates are then incubated in a humidified incubator at 37 ° C and 5% CO2. Cell supernatants are collected 24 h and 48 h later by transferring the cell medium to a 96-well V-bottom polypropylene plate and stored at -80 ° C until reading by ELISA.

3.5.6. IL-6 quantification by ELISA

[386] Levels of IL-6 secreted in cell supernatants are quantified in an enzyme linked immunosorbent assay (ELISA). A 384-well Lumitrac ™ white plate is coated overnight with 40 pL per well of 1 pg / mL mouse human anti-IL-6 antibody solution (R&D Systems, MAB206) diluted in 4 ° PBS Ç. After washing the wells twice with 100 μl of PBST and once with PBS, the remaining binding sites are blocked with 100 μl of blocking buffer (1% BSA and 5% sucrose in PBS) and incubated for 4 h in rt After the blocking step, the wells are washed once with PBST, followed by the addition of samples or recombinant human IL-6 (R&D Systems, 206-IL-050) as standard. The samples are diluted 1/20 in dilution buffer and 40 µl are added for overnight incubation at 0 ° C. The wells are then washed 3 times, twice with PBST and once with PBS, after adding 35 μl of secondary biotinylated anti-IL-6 detection antibody (polyclonal goat antibody biotinylated with human IL-6 (R&D) Systems, BAF206)) at a final concentration of 50 ng / mL. After 2 h incubation in r.t. and appropriate washing steps (twice with PBST and once with PBS), the wells are incubated with 35 pL of a streptavidin-HRP solution diluted 1 / 2,000 (Invitrogen,

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SNN2004), followed by a 45 min incubation in r.t. in the dark. The wells are then washed 3 times (twice with PBST and once with PBS), followed by 5 min of incubation with a 50 µL of substrate solution for chemiluminescence ELISA (Roche, 11 582 950 001). The luminescence of the converted substrate is measured with a Luminoskan ™ Ascent luminometer.

3.5.7. Quantification of MMP13 by ELISA

[387] Levels of MMP13 secreted in cell supernatants are quantified in an antibody capture activity assay. For this purpose, black Nunc® MaxiSorp ™ plates with 384 wells are coated with 35 pL of a solution of anti-MMP13 antibodies at 1.5 pg / mL overnight at 4 ° C. After washing the wells twice with PBST, the remaining binding sites are blocked with 100 µl of 5% skimmed milk in PBS for 24 h at 4 ° C. After the blocking step, the wells are washed twice with PBST, followed by the addition of samples and standards. The samples are diluted 1/5 in dilution buffer and 35 pL are added over 4 h at r.t. The wells are then washed twice with PBST. Subsequently, the MMP13 protein is fully activated by adding 35 pL of a 1.5 mM ΑΡΜΑ solution (SigmaAldrich, A9563) and incubated at 37 ° C for 1 h. The wells are then washed twice with PBST and 35 µl of MMP13 substrate (fluorogenic substrate OMNIMMP® (BIOMOL, P-126)) are added. After incubation for 1 h at 37 ° C, the fluorescence of the converted substrate is measured with a PerkinElmer EnVision® (excitation wavelength: 320 nm, emission wavelength: 405 nm).

3.5.8. Data analysis and calculation

[388] All controls are measured within the linear range of the ELISA standard curve for IL-6 and human MMP13. All data

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111/154 generated are validated against the assay quality parameters (signal / background> 2 and Z '> 0.3).

[389] Unstimulated samples (without activator / vehicle (0.2% DMSO)) are used as a positive control (100% inhibition). As a negative control (0% inhibition), activated samples (activator / vehicle (0.2% DMSO)) are used. The positive and negative controls are used to calculate the Z 'values and the percentage of inhibition (PIN).

RUactivator / vehicle - RUcompound test

PIN = ----------: - ------------------------- X 100

RUactivator / vehicle - Rhemativator / vehicle with RU stands for Relative Chemiluminescence Light Units or relative fluorescence units for IL-6 and MMP13 ELISA, respectively. PIN values are represented for the test compounds tested in the concentration-response and the delCsosion values derived using the Graph Pad Prism® software applying curve adjustment by non-linear (sigmoidal) regression.

Table IX. Results of the SW1353 cell selectivity test of illustrative compounds of the invention

IL-1 β activator TNFa activator Cpd # IL-6 IC50 (nM) [PIN at 20 μΜ] MMP13 ICso (nM) [PIN at 20 μΜ] IL-6 ICso (nM) [PIN at 20 μΜ] MMP13 ICso (nM) [PIN at 20 μΜ] 1 54 [89%] 36 [81%] > 20000 [54%] > 20000 [26%] 12 40 [80%] 29 [75%] > 20000 [22%] > 20000 [36%]

3.5.9. Conclusion

[390] The data in Table IX shows that the compounds according to formula I of the composition of the invention potentially inhibit the expression of IL-6 and MMP13 in IL-1 β activated SW1352 cells, while the effect of such compounds on TNFa-activated events are limited, both in terms of power and maximum amplitude. These data confirm the selectivity of the compounds according to

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112/154 with formula I of the composition of the invention by IRAK-4 controlled pathways, with very limited impact on TNFα signaling which can, in turn, limit the occurrence of treatment-associated side effects, such as neutropenia and infection.

Example 4. ADME assays

4.1. Kinetic solubility

[391] Starting from a 3.3 mM DMSO compound stock solution, a serial dilution of the DMSO compound is prepared by making 1/2: 3.3, 1.6, 0.83, 0 dilutions , 41 and 0.21 mM. This series of dilutions is transferred to a 96-well transparent V-bottom plate (Greiner, 651201) and further diluted 1 / 33.5 in 0.1 M phosphate buffer, pH 7.4, or citrate at 0.1 M, pH 3.0. The final concentrations of the compound are 99.5, 49.7, 24.9, 12.4 and 6.22 μΜ. The final concentration of DMSO does not exceed 3%. As a positive control for precipitation, pyrene (30 mM) is added to the wells on the edge of each 96-well plate. The assay plates are sealed and incubated for 1 h at 37 ° C, shaking at 230 rpm. The plates are then checked under a white light microscope, producing individual images (50x magnification) of the precipitate by concentration. Each well is analyzed by image analysis software and the highest concentration in which the compound appears completely dissolved is recorded.

4.2. Microsomal Stability

[392] A 10 mM stock solution of the compound in DMSO is diluted three times in DMSO. This pre-diluted compound solution is then diluted to 2 M in a 105 mM phosphate buffer (pH 7.4) in a deep 96-well plate (Nunc, 278752) and preheated to 37 ° C.

[393] A glucose-6-phosphate dehydrogenase working solution (G6PDH, Roche, 10127671001) of 700 U / mL is diluted with a

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113/154 factor 1: 700 in a 105 mM phosphate buffer, pH 7.4. A mixture of cofactor containing 0.528 M MgCl ₂ .6H ₂ O (Sigma, M2670), 0.528 M Dglycoside-6-phosphate (Sigma, G7879) and 0.208 M NADP + (Sigma, N0505) is diluted with a factor of 1: 8 in a 105 mM phosphate buffer, pH 7.4.

[394] A working solution is made which contains 1 mg / ml of liver microsomes (Tebu-bio) of the species of interest (for example, human, mouse, rat, dog), 1.2 U / ml of G6PDH and mixture cofactor (6.6 mM MgCl _2, 6.6 mM glucose-6-phosphate, 2.6 mM NADP +). This mixture is pre-incubated for 15 min, but never more than 20 min, in rt

[395] After pre-incubation, the dilution of the compound and the mixture containing the microsomes are added in equal amounts and incubated for 30 min at 300 rpm. For the 0 min time point, two volumes of MeCN are added to the compound dilution before the microsome mixture is added. The final concentrations during incubation are: test compound or control compound at 1 μΜ, 0.2% DMSO, 0.5 mg / ml microsomes, 0.6 U / ml G6PDH, MgCI ₂ at 3.3 mM, 3.3 mM glucose-6-phosphate and 1.3 mM NADP +.

[396] After 30 min of incubation at 37 ° C, the reaction is stopped with 2 volumes of MeCN.

[397] The samples are mixed, centrifuged and the supernatant is collected for analysis in LC-MS / MS. The instrument's responses (ie peak heights) are referenced to the zero time point samples (considered 100%) to determine the percentage of compound remaining. Propranolol and verapamil are included as references in the design of the assay.

[398] Data on microsomal stability are expressed as a percentage of the total amount of compound remaining after 30 min of incubation.

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4.3. Metabolic stability in the S9 subcellular fraction

[399] The purpose of this assay is to evaluate the compound's metabolism by aldehyde oxidase when determining its in vitro metabolic stability in the S9 subcellular fraction.

[400] A stock solution of compound in 10 mM DMSO is first diluted in DMSO (40 times) to obtain a concentration of 250 μΜ. This compound solution is further diluted with water (5 times) to obtain a 50 μΜ working compound solution (to obtain the final compound concentration of 1 μΜ). Hydralazine (selective aldehyde oxidase inhibitor) is prepared in 5 mM water (to obtain a final concentration of 100 μΜ). Mixtures for incubation are prepared by adding 10 μΙ of liver S9 suspension (human, rat, mouse, monkey, BD Gentest ™, 20 mg / ml) to 86 μΙ of 50 mM potassium phosphate buffer, pH 7.4 at 37 ° C (final concentration of 2 mg protein / mL). 2 μΙ_ of 5 mM hydrogenazine is added for incubations with the addition of selective inhibitor or 2 μΙ_ of water for incubations without inhibitor. After 5 min of preheating, the reaction is initiated by adding 2 μΙ_ of test compound at 50 μΜ to the incubation mixture. After 0, 3, 6, 12, 18 and 30 min of incubation, the reaction (100 μΙ_) is terminated with 300 μΙ_ MeCN: MeOH (2: 1) with a mixture of 1% acetic acid containing 10 ng / mL warfarin as an internal analytical standard. The samples are mixed, centrifuged and the supernatant analyzed by LC-MS / MS. Phthalazine is included as a positive control.

[401] Instrument responses (peak compound area ratios and internal standard) are referenced to samples from time zero point (considered 100%) to determine the percentage of compound remaining. Representations of the% of compound remaining are used to determine the intrinsic half-life and clearance in S9 incubations using the GraphPad software

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Prism®. The following formula is used to calculate intrinsic elimination in vitro (pL / min / mg):

CLint (pL / min / mg) = 0.693 / ti / 2 (min) * (ml of incubation / mg of protein) * 1000

[402] Test compounds can be classified as aldehyde oxidase substrates if the elimination of S9 is inhibited by hydralazine. Species specific elimination of the test compound may also indicate metabolism by aldehyde oxidase.

4.4. Metabolic stability in hepatocytes

[403] A stock solution of the test compound in 10 mM DMSO is first diluted in 3 mM DMSO and then in modified Krebs-Henseleit buffer (Sigma, K3753) at 5 μΜ. This compound dilution is added to a suspension of pooled cryopreserved hepatocytes (BioreclamationIVT) at 37 ° C under gentle agitation. The final reaction conditions are: test compound at 1 μΜ, DMSO at 0.03%, 0.5 million viable hepatocytes / mL and an incubation volume of 75 μΙ_. Testosterone (1 μΜ) and 7-hydroxycoumarin (1 μΜ) are used, respectively, as phase I and phase II metabolic reaction controls.

[404] After 0, 10, 20, 45, 90, 120 and 180 min of incubation, the reaction is terminated with 225 μΙ_ of MeCN: MeOH (2: 1) containing 10 ng / mL of warfarin sodium as an internal analytical standard . The samples are mixed, centrifuged and the supernatant analyzed by LCMS / MS.

[405] Instrument responses (test compound proportions and peak areas of the internal standard) are referenced to the zero time point samples (considered 100%) to determine the percentage of compound remaining.

[406] Representations of the percentage of remaining compound are used to determine the half-life and intrinsic elimination in the

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4.5. CYP inhibition

[407] The potential inhibitor of a test compound for human cytochrome P450 isoenzymes (CYP1A2, 2C9, 2C19, 2D6 and 3A4) is assessed using human cytochrome P450 isoenzymes expressed in cDNA and non-fluorescent substrates, which are metabolized to metabolites fluorescent.

[408] The compounds are tested at 3.3 and 10 μΜ, with a final DMSO concentration of 0.3%. The compounds are incubated for 15 min with enzyme before the cofactor / substrate mixture is added. The final reaction concentrations in the cofactor mix for the CYP3A4 (BD Biosciences, 456202), CYP2C9 (BD Biosciences, 456258), CYP2C19 (BD Biosciences, 456259) and CYP1A2 (BD Biosciences, 456203) assays are: 0.4 U / mL glucose-6-phosphate dehydrogenase (G6PDH, Roche, 10165875001), 3.3 mM MgCh (Sigma, M2670), 3.3 mM D-glucose-6-phosphate (Sigma, G7879) and NADP + at 1 , 3 mM (Sigma, NO505). For CYP2D6 (BD Biosciences, 456217), the final reaction concentrations in the assay are 0.4 U / mL G6PDH, 0.41 mM MgCh, 0.41 mM D-glucose-6-phosphate and NADP + 8 , 2 μΜ. The enzyme and substrate concentrations are reported in Table X. After an incubation period, the reaction is stopped by adding a reaction termination solution. For experiments with DBF as a substrate, a 2 N NaOH reaction termination solution is used, while for all other substrates, an 80% MeCN / 20% Tris base 0 reaction termination solution is used. ,5 M.

[409] Fluorescence is read immediately (for CEC, AMMC, BFC) or after 20 min (for CYP2C9 and CYP3A4 using DBF as a substrate) on a PerkinElmer EnVision® reader at the appropriate excitation and emission wavelengths (see Table X) .

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[410] Then, the percentage of CYP inhibition by the test compound is calculated by normalizing the data in blank samples: 100% inhibition is the blank sample stopped before adding the enzyme / substrate mixture and 0% inhibition is the blank sample interrupted after the enzymatic reaction has occurred (50 min).

Table X. Conditions of the inhibition assay used for each CYP450 isoenzyme studied

CYP3A4 CYP3A4 CYP2C19 CYP2C9 CYP1A2 CYP2D6 Substrate (μΜ) DBF CEC AMMC BFC 1 120 35 0.5 4 0.5 Phosphate buffer, pH 7.4 (mM) 200 90 25 25 25 25 Enzyme (pmol / cavity) 1 1.5 6 2 1.5 3 Incubation time (min) 50 50 50 50 50 50 Positive control ketoconazole ketoconazole fluvoxamine sulfaphenazole fluvoxamine quinidine Excitation wavelength (nm) 485 400 400 485 400 380 Emission wavelength (nm) 530 530 460 530 460 460

411] AMMC: aminoethyl-7-methoxy-4-methylcoumarin

[412] CEC: 3-cyano-7-ethoxycoumarin

[413] BFC: 7-benzyloxy-4-trifluoromethylcoumarin

[414] DBF: dibenzylfluorescein

4.6. Permeability in MDCKII-MDR1

[415] MDCKII-MDR1 cells are Madin-Darby canine kidney epithelial cells that overexpress the human multi-drug resistance gene (MDR-1) encoding Pglicoprotein (P-gp). The cells are obtained from the Neterlands Cancer Institute and used after a 3-4 day culture in plates with 24 well Millicell® cell culture insert (Millipore,

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PSRP010R5). A permeability assay on bidirectional MDCKII-MDR1 is performed as described below.

[416] 3 χ 10 ⁵ cells / mL (1.2 χ 10 ⁵ cells / well) were seeded in culture medium consisting of DMEM (Sigma, D5796) + 1% Glutamax-100 (Sigma, G8541) + antibiotic / 1% antimycotic (Sigma, A5955) + 10% FBS (Sigma, F7524; inactivated at 56 ° C for 30 min). The cells are left in the CO2 incubator for 3-4 days. The medium is changed 24 h after culture and on the day of the experiment.

[417] Test and reference compounds (Amprenavir (Moravek Biochemicals, M-1613), diclofenac (Sigma, D6889)) are prepared in Dulbecco's phosphate buffered saline (D PBS, pH 7.4; Sigma, D8662 ) and added to the apical (400 μΙ_) or basolateral (800 μΙ_) chambers of the Millicell cell culture plate set at a final concentration of 10 μΜ (0.5 μΜ in the case of Amprenavir) with a final concentration of DMSO at 1 %. A recipient solution (D-PBS + 1% DMSO) is added to the opposite chamber of the Millicell cell culture plate.

[418] Lucifer yellow at 100 μΜ (Sigma, L0259) is added to all donor buffer solutions in order to assess the integrity of the cell monolayers when monitoring Lucifer yellow permeation. Lucifer yellow is a fluorescent marker for the paracellular transport pathway and is used as an internal control to verify the integrity of the narrow junction of each cell monolayer during the assay.

[419] After 1 h of incubation at 37 ° C while shaking on an orbital shaker at 150 rpm, 75 pL aliquots are removed from both apical and basal chambers and added to 225 pL of MeCN solution: water (2: 1 ) containing an internal analytical standard (10 ng / mL of warfarin) in a 96-well plate. The rate is also

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119/154 used at the beginning of the experiment from donor solutions to obtain initial concentrations.

[420] The concentration of the compound in the samples is measured using high performance liquid chromatography / mass spectroscopy (LC-MS / MS).

[421] Lucifer yellow is measured with a Thermo Scientific Fluoroskan Ascent FL (excitation wavelength: 485nm, measurement wavelength: 530 nm) in a 96-well plate containing 150 μΙ_ of liquid from all recipient wells (basolateral or apical side).

Example 5. Whole blood assays

5.1. Inhibition of human TNF-α release ex vivo (whole blood assay)

[422] The purpose of the assay is to evaluate the activity of the compounds of the invention in the TLR / IRAK-4 pathway activated in an ex vivo human whole blood environment. Toll-like receptors (TLRs) are pattern recognition receptors that recognize a wide variety of microbial molecules, called pathogen-associated molecular patterns (MMAPs). Human TLR7 and TLR8 recognize imidazoquinoline compounds (eg, CL097) and single stranded RNAs as their natural ligands. Activation of TLRs leads to the production of various cytokines (eg, TNFα, IL-8, IL-6) by cells treated with a TLR agonist. Cytokine release is used as a reading in this assay and represents a measure of the level of inhibition of the TLR / IRAK-4 pathway by the tested compound. It should be noted that, in the context of a complete organism, there are other sources for these cytokines that are not dependent on the TLR / IRAK-4 pathway such as, for example, macrophages (after activation of the Fcy receptor) (Yan et al., 2012)) or T cells (upon activation of the T cell receptor (Brehm etal., 2005)).

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5.1.1. Experimental design

[423] Blood is collected from healthy volunteers in lithium heparin tubes by venipuncture and then gently inverted several times to prevent clotting and incubated for at least 15 min at 37 ° C on an oscillating shaker / mixer. Then, 200 pL of blood are distributed in 2 mL microtubes and pre-incubated in duplicate with 0.3% DMSO or test compound in different concentrations (10 to 0.01 μΜ, 3 dilutions in RPMI 1640 medium without glutamine (Life Technologies , 31870)) for 15 min at 37 ° C. After this pre-incubation, the blood is activated with CL097 (2 pg / mL from a 1 mg / mL solution in water; InvivoGen, tlrl-c97) or vehicle (distilled water) for 3 h 30 min at 37 ° C . The microtubes are centrifuged at 5000 χ g for 10 min at 4 ° C and approximately 80 pL of plasma are collected in a 96-well polystyrene plate. The plasma can be analyzed fresh or frozen at 80 ° C right after activation. Finally, TNFa quantification is performed by diluting the plasma 40 times using the TNF-alpha DuoSet ELISA kit (R&D Systems, DY210), according to the manufacturer's instructions. Optical density (OD) is determined at 450 nm in a PerkinElmer EnVision 2102 Multilabel plate reader.

5.1.2. Data analysis

[424] The standard curve is created by plotting the average absorbance on the y-axis graphically as a function of the concentration on the x-axis and a better fit of the curve is plotted through the points on the graph. A linear regression analysis is performed to determine the equation (y = ax + b) and the value of R-square. For each replicated blood sample, the concentration of TNFa is calculated taking into account the dilution factor using the formula:

TNFa concentration _amos trai = 40 * (OD _amos tra 1 - b) / a

[425] The data is then expressed as a percentage

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121/154 inhibition (PIN) for each replica using the formula:

ΤΜΡα mêdso with CLÕ97 - TSFa sample 1

Sample PIN 1 = -----—--------------—-----; —— x 1QG, casn CLG97 -T ^ Fsx medsa cm veieuw where average TNFa with CL097 is the average concentration of TNFα from replicated samples activated with CL097; TNFa sample 1 is the TNFa concentration of sample 1; and mean vehicle TNFa is the average TNFa concentration of replicated samples treated with vehicle.

[426] Curve adjustments are generated using the mean ± SEM PIN. Derived IC graphs and calculations using GraphPad Prism® software ·

5.2. Inhibition of TNF-α release in rat ex vivo (whole blood assay)

[427] The purpose of the assay is to evaluate the activity of the compounds of the invention in the TLR / IRAK-4 pathway activated in an ex vivo rat whole blood environment. Toll-like receptors (TLRs) are pattern recognition receptors that recognize a wide variety of microbial molecules, called pathogen-associated molecular patterns (MMAPs). Although human TLR7 and TLR8 recognize imidazoquinoline compounds (eg, CL097) and single stranded RNAs as their natural ligands, rodent TLR8 needs additional factors, such as oligodeoxynucleotides (eg, poly (dT)) for activation.

5.2.1. Experimental design

[428] Sprague-Dawley rats (males, 7-8 weeks old, body weight 200-250 g) are obtained from Janvier Labs (France).

[429] Blood, obtained by exsanguination, is collected from at least 2 rats in lithium heparinate tubes and then pre-incubated for at least 15 min at 37 ° C on an oscillating shaker / mixer. All rats' blood is mixed in a 50 mL polypropylene tube to obtain a single blood batch.

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122/154 co. Then, 200 qL of blood are distributed in 2 ml microtubes and incubated in duplicate with 0.3% DMSO or test compound in different concentrations (from 10 to 0.01, 3 dilutions in RPMI 1640 medium without glutamine (Life Technologies, 31870)) for 15 min at 37 ° C. After this pre-incubation, the blood is activated with CL097 (10 pg / mL from 1 mg / mL solution in water; InvivoGen, tlrl-c97) and poly (dT) (1 μΜ from solution in water at 100 μΜ; InvivoGen, tlr1-pt17) or vehicle (distilled water) for 3 h 30 min at 37 ° C. The microtubes are centrifuged at 5000 χ g for 10 min at 4 ° C and about 80 qL of plasma are collected in a 96-well polystyrene plate. Plasma can be analyzed fresh or frozen at - 80 ° C right after activation. Finally, quantification of TNFa is performed in plasma (diluted 1: 3) using the TNF-alpha Quantikine ELISA kit for mice (R&D Systems, SRTA00), according to the manufacturer's instructions. Optical density (OD) is determined at 450 nm in a PerkinElmer EnVision 2102 Multilabel plate reader.

5.2.2. Data analysis

[430] The standard curve is created by plotting the average absorbance on the y axis graphically as a function of the concentration on the x axis and a better fit of the curve is plotted through the points on the graph. A linear regression analysis is performed to determine the equation (y = ax + b) and the value of R-square. For each replicated blood sample, the concentration of TNFa is calculated taking into account the dilution factor using the formula:

TNFa concentration _amO strai = 40 * (OD _amO stra 1 - b) / a

[431] The data is then expressed as a percentage of inhibition (PIN) for each replica using the formula:

TKFct medium cam CL097 - TNF «sample 1

Sample PIN i = ----------------------------------- x 10G,

TNFa measured with CLÜ97 medsa with vehicle where average TNFa with CL097 is the average TNFa concentration of replicated samples activated with CL097 + poly (dT); TNFa sample 1

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123/154 is the TNFa concentration in sample 1; and mean vehicle TNFa is the average TNFa concentration of replicated samples treated with vehicle.

[432] Curve adjustments are generated using the mean PIN ± SEM. IC graphs and calculations derived using GraphPad Prism® software.

Example 6. In vivo tests

6.1. A model of psoriatic epidermal hyperplasia in mice induced by topical applications of imiquimod, a TLR7 / 8 agonist.

6.1.1. Materials

[433] Aldara® 5% imiquimod cream is obtained from MEDA.

[434] The purified mouse anti-IL-12 / IL-23 antibody p40 FG (C17.8) is obtained from Affymetrix eBioscience (catalog number 16-7123-85).

6.1.2. Animals

[435] Balb / cJ mice (females, body weight 18-20 g) are obtained from Janvier Labs (France). The mice are kept on a 12-hour light / dark cycle (07: 00-19: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.1.3. Study design

[436] The study design is adapted from Van der Fits L. et al. (van der Fits etal., 2009).

[437] On the first day, the mice are shaved around both ears under light anesthesia with isoflurane.

[438] 30 mg of commercially available imiquimod cream (5% Aldara cream) are applied to the inner and outer surfaces of each ear for 4 consecutive days, that is, in a daily dose of

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1.5 mg of the active compound. Control animals received the same amount of petroleum jelly.

[439] From day 1 to day 5, mice are dosed with the test compound, 10 or 30 mg / kg, po, bid, in 0.5% methylcellulose, before imiquimod application (on day 5, mice are dosed only once, 2 h before euthanasia).

[440] In a positive reference group, animals received two intraperitoneal injections of mouse anti-IL-12 / IL-23 antibody p40, 10 mg / kg, on day 1 and 3 days before day 1.

6.1.4. Disease assessment

[441] The thickness of both ears is measured daily with a thickness gauge (Mitutoyo, Absolute Digimatic, 547321). Body weight is assessed at the beginning of the experiment and during euthanasia. On day 5, 2 h after the last dosage, the mice are sacrificed. The ear pavilions are cut, excluding the cartilage. The pavilions are weighed and then immersed in a flask containing 1 mL of RNA / ater® solution to assess gene expression or formalin for histology.

[442] There are 14 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using a unidirectional ANOVA, followed by Dunnett's post-hoc test against the imiquimodode vehicle group.

6.1.5. Histology

[443] After euthanasia, the ears are collected and fixed in 3.7% formaldehyde before being included in paraffin. 2 qm thick sections are cut and stained with hematoxylin and eosin. The thickness of the epidermis of the ear is measured through image analysis (SisNcom software) with 6 images per ear captured at 20x magnification. Data are expressed as mean ± SEM and statistical analysis is performed using a one-way ANOVA

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125/154 nal, followed by Dunnett's post-hoc test against the imiquimode-vehicle group.

6.1.6. Gene expression analysis

[444] The ears are removed from the RNA / ater® solution and placed in Trizol® after rupture with 1.4 mm ceramic beads in a Precellys device. The total RNA is then purified using the NucleoSpin® RNA kit. cDNA is prepared and quantitative PCR is performed with specific primers for Qiagen genes using SYBR Green technology in a ViiA7 real-time PCR system (Applied Biosystems). The expression levels of each gene (IL17A, IL1, IL22, LCN2, S100A8 and S100A9) are calculated in relation to the expression level of the cyclophilin A gene. Data are expressed as mean ± SEM of the relative quantity (RQ = 2 ^ac t, where ACt = Ct sample - Orda cyclofilina). The statistical test used is ANOVA analysis of variance with Dunnett's post-hoc test against the vehicle imiquimod group.

6.2. Model of psoriatic epidermal hyperplasia in mice induced by intradermal injections of IL-23

6.2.1. Materials

[445] Recombinant, carrier-free mouse IL-23 (14-8231, CF) is provided by e-Bioscience.

6.2.2. Animals

[446] Balb / c mice (females, body weight 18-20 g) are obtained from CERJ (France). The mice are kept on a 12-hour light / dark cycle (07: 00-19: 00). The temperature is maintained at 22 ° C, food and water are provided ad libitum.

6.2.3. Study design

[447] The study design is adapted from Rizzo H.L. et al. (Rizzo etal., 2011).

[448] On the first day (D1), the mice are shaved in

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[449] For 4 consecutive days (D1 to D4), mice received a daily intradermal dose of recombinant mouse IL-23 (1 pg / 20 μΙ_ in PBS / 0.1% BSA) in the right ear pavilion and 20 pL of PBS / 0.1% BSA in the left ear pavilion under anesthesia induced by isoflurane inhalation.

[450] From D1 to D5, mice are dosed with test compound (10, 30 or 100 mg / kg, po, qd, in 0.5% methylcellulose) or with vehicle, 1 h before the injection of IL- 23.

6.2.4. Disease assessment

[451] The thickness of both ears is measured daily with an automatic calibrator. Body weight is assessed at the beginning and during euthanasia. On the fifth day, 2 h after the last dosage, the mice are sacrificed. The ear pavilions are cut, excluding cartilage. The pavilions are weighed and then placed in a flask containing 1 ml of RNA / ater® solution or in formaldehyde.

[452] At D4, blood samples are also collected from the retro-orbital sinus to characterize the PK profile immediately before dosing (T0) and 1 h, 3 h, 6 h post-dosing.

[453] There are 8 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using unidirectional ANOVA followed by Dunnett's post-hoc test versus vehicle-IL-23 groups.

6.2.5. Histology

[454] After euthanasia, the ears are collected and fixed in 3.7% formaldehyde before being included in paraffin. 2 pm thick sections are made and stained with hematoxylin and eosin. The thickness of the epidermis of the ear is measured through image analysis (Sis'Ncom software) with 6 images per ear captured

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127/154 with x20 magnification. Data are expressed as mean ± SEM and statistical analysis is performed using unidirectional ANOVA, followed by Dunnett's post-hoc test versus vehicle-IL-23 groups.

6.2.6. Gene expression analysis

[455] Half of the ears are removed from the RNA / ater® solution and placed in Trizol® after rupture with 1.4 mm ceramic beads in a Precellys device. The total RNA is then purified using a NucleoSpin® RNA kit. CDNA is prepared and quantitative PCR is performed with Qiagen gene specific primers using SYBR Green technology in a ViiA7 real-time PCR system (Applied Biosystems). The expression levels of each gene (IL17A, IL1B, IL22, LCN2, S100A8 and S100A9) are calculated in relation to the expression level of the cyclophilin A gene. Data are expressed as mean ± SEM of the relative quantity (RQ = 2 ^ac t, where ACt = Ct sample - Orda cyclofilina). The statistical test used is ANOVA analysis of variance with Dunnett's post-hoc test against the vehicle-IL-23 group.

6.3. PK / PD model: CL097-induced TNF-α release, a specific TLR7 / 8 agonist

[456] The purpose of this assay is to determine the relationship between inhibition of an IRAK-4 dependent event in vivo by administering a compound of the invention and the circulating concentration levels of this compound.

6.3.1. Materials

[457] CL097 (cat. No. Tlrl-C97) and poly (dT) (cat. No. Tlrl-PT17) are obtained from InvivoGen.

[458] AlphaLISA® mouse TNF-α kits are obtained from Perkin-Elmer (cat. No. AL505C).

6.3.2. Animals

[459] DBA / 1J mice (males, body weight 18-20 g)

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128/154 are obtained from Janvier Labs (France). The mice are kept on a 12-hour light / dark cycle (07: 00-19: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.3.3. Study design

[460] The mice receive an oral dose of the test compound. A group of intact animals that receive no dosage is used as the time point = 0.

[461] Two blood samples obtained by intracardiac sampling (under anesthesia with isoflurane) are collected in lithium heparin tubes at 30 min, 1 h, 3 h, 8 h or 24 h after dosing. One is used for pharmacokinetic analysis (PK) and the second for quantification of pharmacodynamic markers (PD).

6.3.4. Quantification of plasma levels of compounds

[462] Whole blood samples are centrifuged at 5,000 rpm for 10 min and the resulting plasma samples are stored at 20 ° C, awaiting analysis. The plasma concentrations of each test compound are determined using an LCMS / MS method.

6.3.5. Determination of pharmacokinetic parameters

[463] Pharmacokinetic parameters are calculated using WinNonlin® (Pharsight®, United States).

6.3.6. Quantification of PD marker

[464] Each blood sample is stimulated with CL097 and poly (dT) for 2 h at 37 ° C. Then, the plasma is collected and analyzed for TNF-α by AlphaLISA, according to the manufacturer's instructions.

[465] There are 6 mice per group. The results are expressed as TNF-α concentration (pg / mL) or as the percentage of inhibition (PIN) in relation to time point t = 0. Data are displayed

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129/154 seated as mean ± SEM and statistical analysis is performed using unidirectional ANOVA, followed by Dunnett's post-hoc test versus the vehicle group at the corresponding time point.

6.4. Prophylactic model of atopic dermatitis in mice induced by topical application of MC903

6.4.1. Materials

[466] 0.5% methylcellulose is obtained from VWR (cat. No. AX021233). MC903 (calcipotriol) is obtained from Tocris Bioscience (cat. No. 2700/50). ProSense®680 is obtained from PerkinElmer (cat. No. NEV10003). RNA / ater® is obtained from Ambion (cat. No. AM7021). Imalgene® 1000 (Merial) and Rompun®2% (Bayer) are obtained from Centravet (cat. No. IMA004-6827812 and ROM001-6835444).

6.4.2. Animals

[467] BALB / cN mice (females, body weight 1820 g) or CD1 / Swiss mice (females, body weight 2426 g) are obtained from Janvier Labs (France). The mice are kept on a 12-hour light / dark cycle (07: 0019: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.4.3. Study design

[468] The study design is adapted from Li M. et al. (Li et al., 2006).

[469] On the first day (D1), the mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g) and scraped around both ears.

[470] From the D1,20 pL of EtOH or 2 nmol of MC903 (in 20 pL of EtOH) they are applied topically to both ears of the mice for five consecutive days.

[471] From D1 to D8, mice are dosed with test compound (15 or 30 mg / kg, p.o., b.i.d., in 0.5% methylcellulose) or

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130/154 xamethasone (5 mg / kg, p.o., q.d., in 0.5% methylcellulose) or vehicle.

6.4.4. Quantification of plasma levels of compounds

[472] The plasma concentrations of each test compound are determined using an LC-MS / MS method in which the mass spectrometer is operated in positive or negative electrospray mode.

6.4.5. Determination of pharmacokinetic parameters

[473] Pharmacokinetic parameters are calculated using Phoenix®WinNonlin® (Pharsight®, United States).

6.4.6. Disease assessment

[474] The thickness of both ears is measured (after anesthesia induced by isoflurane inhalation) at the beginning of the study, on alternate days and in euthanasia using a thickness gauge (Mitutoyo, Absolute Digimatic, 547-321).

[475] Body weight is assessed at baseline, on alternate days and during euthanasia.

[476] At D4, mice in all groups received the ProSense®680 probe (0.8 nmol / 10 g, IP). At D5, mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g). Granulocyte infiltration is measured using molecular imaging in vivo (Bruker In-Vivo Xtreme imaging system, excitation wavelength: 630 nm, emission wavelength: 700 nm, acquisition time: 5 seconds).

[477] At D8, 2 h after the last dosage, mice are sacrificed and whole blood is collected in tubes coated with EDTA and the plasma is frozen for further measurements (including the circulating compound). A blood sample is also collected in tubes coated with heparin.

[478] The ear pavilions are collected and weighed. An

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131/154 ear is cut longitudinally in two halves. Half is fixed in 4% formaldehyde for histology; the other is immersed in RNA / ater® to evaluate gene expression.

[479] There are 8 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using one-way ANOVA, followed by Dunnett's post-hoc test versus groups with MC903 vehicle for ear thickness and weight, versus group with EtOH vehicle for body weight.

6.4.7. Histology

[480] After euthanasia, half of the ears are collected and fixed in 3.7% formaldehyde before being included in paraffin. Sections 4 μιτι thick are immunostained by immunohistochemistry with cell specific marker antibody: CD3 for T cells and EPX for eosinophils. The immunostained cell areas of an entire section per mouse are measured using image analysis (CaloPix software, TRIBVN Healthcare). Data are expressed as mean ± SEM and statistical analysis is performed using one-way ANOVA, followed by Dunnett's post-hoc test versus vehicle-MC903 groups.

6.4.8. Gene expression analysis

[481] The ears are removed from the RNA / ater® solution and placed in Trizol® after rupture with 1.4 mm ceramic beads in a Bertelly Instruments Precellys® homogenizer. The total RNA is then extracted using a phenol / chloroform protocol and purified with a QIAcube using an RNeasy® 96 QIAcube® HT kit (Qiagen, cat. No. 74171). cDNA is prepared and quantitative PCR is performed with specific primers for Qiagen gene using SYBR Green technology in a ViiA 7 real-time PCR system (Applied Biosystems). The expression levels of each gene (IL4, IL5, IL13, TSLP, IL33, ST2, IL25, IL31, IFN γ, IL6, IL10, LCN2, S100A8 and

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S100A9) are calculated in relation to the expression levels of the housekeeping gene HPRT, GAPDH and β-actin. The data are expressed as mean ± SEM of the relative quantity (RQ = 2 ^ac t, where ACt = Ct sample - mean (Ct for HPRT, Ct for GAPDH, Ct for βactin). The statistical test used is ANOVA analysis of variance with Dunnett's post-hoc test versus the EtOH / MC903 vehicle group.

6.5. Therapeutic model of atopic dermatitis in mice induced by topical application of MC903

6.5.1. Materials

[482] 0.5% methylcellulose is obtained from VWR (cat. No. AX021233). MC903 (calcipotriol) is obtained from Tocris Bioscience (cat. No. 2700/50). ProSense®680 is obtained from PerkinEImer (cat. No. NEV10003). RNA / ate / ® is obtained from Ambion (cat. No. AM7021). Imalgene® 1000 (Merial) and Rompun®2% (Bayer) are obtained from Centravet (cat. No. IMA004-6827812 and ROM0016835444).

6.5.2. Animals

[483] BALB / cN mice (females, body weight 1820 g) or CD1 / Swiss mice (females, body weight 2426 g) are obtained from Janvier Labs (France). The mice are kept on a 12-hour light / dark cycle (07: 0019: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.5.3. Study design

[484] The study design is adapted from Li M. et al. (Li et al., 2006).

[485] On the first day (D1), the mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g) and scraped around both ears.

[486] From the D1,20 pL of EtOH or 2 nmol of MC903 (in 20

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133/154 μΙ_ of EtOH) are applied topically to both ears of mice until D9, D11 or D15 (except during the weekend).

[487] From D5, mice are dosed with test compound (15 or 30 mg / kg, po, bid, in 0.5% methylcellulose) or dexamethasone (5 mg / kg, po, qd, in methylcellulose 0.5%), or with vehicle, up to D10, D12 or D16.

6.5.4. Quantification of plasma levels of compounds

[488] The plasma concentrations of each test compound are determined using an LC-MS / MS method, in which the mass spectrometer is operated in positive or negative electrospray mode.

6.5.5. Determination of pharmacokinetic parameters

[489] Pharmacokinetic parameters are calculated using Phoenix®WinNonlin® (Pharsight®, United States).

6.5.6. Disease assessment

[490] The thickness of both ears is measured (after anesthesia induced by isoflurane inhalation), before the application of MC903, at the beginning of the study, three times a week and in euthanasia using a thickness gauge (Mitutoyo, Absolute Digimatic, 547-321).

[491] Body weight is assessed at baseline, three times a week and during euthanasia.

[492] In D8, D10 or D11, mice from all groups received the ProSense®680 probe (0.8 nmol / 10 g, IP). The next day (D9, D11 or D12), the mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 ml_ / 10g). The infiltration of granulocytes is then measured using in vivo molecular imaging (Bruker In-Vivo Xtreme imaging system, excitation wavelength: 630 nm, emission wavelength: 700 nm, acquisition time: 5 seconds).

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[493] At D10, D12 or D16, 2 h after the last dosage, the mice are sacrificed; whole blood is collected in tubes coated with EDTA and the plasma is frozen for further measurements (including circulating compound).

[494] Ear pavilions are collected. One ear is cut longitudinally into two halves. Half is fixed in 4% formaldehyde for histology; the other is immersed in RNA / ater® to evaluate gene expression.

[495] There are 8 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using one-way ANOVA, followed by Dunnett's post-hoc test versus groups with MC903 vehicle for ear thickness and weight, versus group with EtOH vehicle for body weight.

6.5.7. Histology

[496] After euthanasia, half of the ears are collected and fixed in 3.7% formaldehyde before being included in paraffin. 4 pm thick sections are immunostained by immunohistochemistry with anti-CD3 antibody. The areas of immunostained cells in an entire section per mouse are measured using image analysis (CaloPix software, TRIBVN Healthcare). Data are expressed as mean ± SEM and statistical analysis is performed using one-way ANOVA, followed by Dunnett's post-hoc test versus groups with MC903 vehicle.

6.5.8. Gene expression analysis

[497] The ears are removed from the RNAlatei® solution and placed in Trizol® after rupture with 1.4 mm ceramic spheres in a Bertelly Instruments Precellys® homogenizer. The total RNA is then extracted using a phenol / chloroform protocol and purified with a QIAcube using an RNeasy® 96 QIAcube® HT kit (Qiagen, cat. No. 74171). cDNA was prepared and quantitative PCR is performed

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135/154 with specific primers for Qiagen gene using SYBR Green technology in a ViiA 7 real-time PCR system (Applied Biosystems). The expression levels of each gene of interest (GOI = IL4, IL5, IL13, TSLP, IL33, ST2, IL25, IL31, IFN-γ, IL6, IL10, LCN2, S100A8 and S100A9) are calculated in relation to the expression levels of the housekeeping gene of HPRT, GAPDH and β-actin.

[498] All qPCR data are expressed as mean ± SEM of the normalized relative quantity (NRQ = 2 ^A (ACq GOI) / Geometric mean (2 ^A (ACq HPRT), 2 ^A (ACq GAPDH), 2 ^A (ACq β -actin)), where ACq = mean Cq - Cq sample The statistical test used is ANOVA analysis of variance with Dunnett's post-hoc test versus the EtOH / MC903 vehicle group.

6.6. Model of systemic lupus erythematosus in mice induced by epicutaneous applications of imiquimod

6.6.1. Materials

[499] Aldara® 5% imiquimod cream is obtained from MEDA.

[500] ELISA kits for double-stranded anti-DNA antibodies are obtained from Alpha Diagnostic International (cat. No. 5120). ELISA kits for mouse urinary albumin are obtained from Abeam (cat. No. Ab108792). Kits for urine creatinine assays are obtained from Abnova (cat. No. KA4344).

6.6.2. Animals

[501] BALB / cJ mice (females, body weight 1820 g) are obtained from Janvier Labs (France). The mice are kept on a 12-hour light / dark cycle (07: 0019: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.6.3. Study design

[502] The study design is adapted from Yokogawa M. et

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136/154 al. (Yokogawa etal., 2014).

[503] On the first day (D1), the mice are shaved around the right ears.

[504] The mice receive an epicutaneous application of 1.25 mg of imiquimod 3 times a week in the right ear pavilion for 12 consecutive weeks (D1 to D86). The control group receives the same amount of petroleum jelly.

[505] From D1 to D86, mice are dosed with the test compound (30 mg / kg, p.o., q.d., in 0.5% methylcellulose) or with vehicle (10 mL / kg).

6.6.4. Disease assessment

[506] The thickness of the ears is measured once a week with an automatic meter (Mitutoyo, Absolute Digimatic, 547-321).

[507] Body weight is assessed at baseline and once a week until euthanasia. At necropsy, the weight of the spleen is also measured. The mice are sacrificed 2 h after the last dosage.

[508] At different times (for example, on days D28, D56 and D84), mice are placed individually in a metabolic cage to perform urinalysis and assess proteinuria (albumin / creatinine ratio).

[509] Sera are collected at different times (for example, at D28, D56 and D86) to assess levels of anti-double stranded DNA IgG.

[510] At D13, blood samples are also collected from the retro-orbital sinus for PK profile characterization just before dosing (T0) and 1 h, 3 h, 6 h post-dosing.

[511] There are 8-19 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using one-way ANOVA, followed by Dunnett's post-hoc test versus vehicle-imiquimod groups.

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6.6.5. Quantification of plasma levels of compounds

[512] The plasma concentrations of each test compound are determined using an LC-MS / MS method, in which the mass spectrometer is operated in positive or negative electrospray mode.

6.6.6. Determination of pharmacokinetic parameters

[513] Pharmacokinetic parameters are calculated using Phoenix®WinNonlin® (Pharsight®, United States).

6.6.7. Histology

[514] After euthanasia, the left kidneys are collected and cut longitudinally into two parts. A portion is fixed in 3.7% formaldehyde before being embedded in paraffin. Sections are 4 pm thick and stained with periodic acid from Schiff (PAS) or immunostained with CD3 (T cells), CD20 (B cells) and F4 / 80 (macrophages).

6.6.7.1. Histopathology

[515] In each glomerulus, 4 different readings, including mesangioproliferation, endocapillary proliferation, expansion of the mesangial matrix and segmental sclerosis, are rated on a scale of 0 to 2 and then added together. For each kidney, about 50 glomeruli are scored and then the average is a glomerular lesion score (Yokogawa etal., 2014). Data are expressed as mean ± SEM and statistical analysis is performed using the KruskalWallis test, followed by the Dunnett post-hoc test versus the vehicle-imiquimod group.

6.6.7.2. Cell quantifications

[516] For each cell type, immunohistochemical analysis is performed using image analysis (CaloPix software, TRIBVN Healthcare) in the entire tissue section with an x20 magnification. Data are expressed as mean ± SEM and statistical analysis is performed

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138/154 using unidirectional ANOVA, followed by Dunnett's post-hoc test versus vehicle-imiquimod group.

6.6.7.3. Gene expression analysis

[517] In euthanasia, the second part of the left kidney is placed in tubes containing 1.4 mm ceramic spheres and broken in RLT lysis buffer with 1% DTT (Qiagen, cat. No. 79216) with a Bertelly homogenizer Instruments Precellys®. The total RNA is then purified with a QIAcube using an RNeasy ®96 QIAcube ® Kit HT (Qiagen, cat. No. 74171). cDNA was prepared and quantitative PCR is performed with specific primers for Qiagen gene using SYBR Green technology in a ViiA 7 real-time PCR system (Applied Biosystems). The expression levels of each gene of interest (GOI = CD3, CD68, CD20, OAS1, Mx1, IFIT1, CXCL11 and Usp18) are calculated in relation to cyclophilin, GAPDH and the expression levels of the β-actin housekeeping gene.

[518] In euthanasia, one third of the spleen is placed in tubes containing 1.4 mm ceramic spheres and ruptured in Trizol® with a Bertelly Instruments Precellys® homogenizer. Total RNA is extracted using a phenol / chloroform process and then purified with a QIAcube using an RNeasy® 96 QIAcube® HT kit (Qiagen, cat. No. 74171). cDNA is prepared and quantitative PCR is performed with Qiagen gene specific primers using SYBR Green technology in a ViiA 7 real-time PCR system (Applied Biosystems). The expression levels of each gene of interest (GOI = CD20, IRF7, OAS1, MX1, IFIT1, CXCL11, Usp18, BCL6, CXCL13, CXCR5, MAF, ICOSL, PDCD1, SH2D1a) are calculated in relation to the expression levels of genes housekeeping of cyclophilin, GAPDH and β-actin.

[519] All qPCR data are expressed as mean ± SEM of the normalized relative quantity (NRQ = 2 ^A (ACq GOI) / Average

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139/154 geometric (2 ^A (ACq for cyclophilin), 2 ^A (ACq for GAPDH), 2 ^A (ACq for β-actin), where ACq = average Cq - sample Cq. The statistical test used is ANOVA analysis of variance with Dunnett's post-hoc test versus the vehicle-imiquimod group.

6.7. Murine-induced psoriatic arthritis model induced by IL-23 overexpression

6.7.1. Materials

[520] The mouse IL-23 episomic overexpression vector (EEV) is obtained from System Biosciences (cat. No. EEV651A-1). Ringer's solution tablets are obtained from Sigma-Aldrich (cat. No. 96724-100TAB). Mouse IL-23 Quantikine ELISA kits are obtained from R&D Systems (cat. No. M2300). ProSense®680 and OsteoSense®750EX are obtained from PerkinElmer (cat. No. NEV10003 and NEV10053EX). RNA / ater® is obtained from Ambion (cat. No. AM7021). Imalgene® 1000 (Merial) and Rompun®2% (Bayer) are obtained from Centravet (cat. No. IMA0046827812 and ROM001-6835444).

6.7.2. Animals

[521] B10.RIII mice (male, 8 weeks old) are obtained from Charles River (France). The mice are kept on a 12-hour light / dark cycle (07: 00-19: 00). The temperature is maintained at 22 ± 2 ° C, food and water are provided ad libitum.

6.7.3. Study design

[522] The study design is adapted from Sherlock J.P. et al. (Sherlock et al., 2012).

[523] On the first day (D1), the mice underwent a hydrodynamic injection of Ringer or IL-23 EEV into Ringer in the tail vein (3 pg / 2.1 mL, IV injected over a period of 4-6 seconds ).

[524] From D5, twice a week, mice

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140/154 are punctuated by clinical symptoms until the end of the experiment.

[525] At D5, blood is collected by puncture into the submandibular vein to assess the serum concentration of IL-23.

[526] At D9, mice from all groups received the ProSense®680 probe (0.8 nmol / 10 g, IP). At D10, the mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g). Granulocyte infiltration is then measured using in vivo molecular imaging (Bruker In-Vivo Xtreme imaging system, excitation wavelength: 630 nm, emission wavelength: 700 nm, acquisition time: 5 seconds).

[527] At D11, randomization is performed according to the design and molecular score of ProSense®680.

[528] From D12, mice are dosed with the test compound (30 mg / kg, p.o., b.i.d. in 0.5% methylcellulose) or with vehicle.

[529] At D19, blood is collected at times T0, T1h, T3h and T6h after the last dose. The plasma is separated and maintained at 20 ° C until bioanalysis.

[530] At D36, mice in all groups are sacrificed 2 h after the last administration of the compound. The following is collected: the heels around the enthesion (without skin) of the left hind limb are immediately frozen in Precellys tubes. The fingers are collected in tubes that contain RNA / ater®. The right posterior limb is immediately fixed in 4% formaldehyde for histological evaluation. X-ray measurement is performed 48 hours after fixation.

[531] An ear is collected in a tube containing RNA / ater® for analysis of transcripts.

[532] Whole blood is collected in a serum blood tube and

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141/154 mixed by gentle inversion 8-10 times. After coagulation, blood samples are centrifuged 10 min at 1800 χ g. After centrifugation, the serum is stored at -80 ° C.

[533] Part of the colon (distal colon 1 cm) is immediately frozen in the Precellys tube for transcript analysis. Another part (distal colon 1 cm) is immediately fixed in 4% formaldehyde for later histological analysis.

6.7.4. Disease assessment

[534] Body weight is assessed at baseline, then twice a week and at euthanasia.

[535] Twice a week, clinical signs of inflammation are scored: 0 for normal paw; 1 if swelling of a finger; 2 if swelling of two or more fingers; 3 if swelling of the entire paw. The scores of all members are added together to produce an overall score.

[536] At D23, mice from all groups received the ProSense®680 probe (0.8 nmol / 10 g, IP). At D24, mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g). Granulocyte infiltration is then measured using in vivo molecular imaging (Bruker In-Vivo Xtreme imaging system, excitation wavelength: 630 nm, emission wavelength: 700 nm, acquisition time: 5 seconds).

[537] At D32, mice in all groups received the ProSense®680 probe (0.8 nmol / 10 g, IP) and the OsteoSense®750EX probe (0.8 nmol / 10 g, IP). At D33, mice are anesthetized with an intraperitoneal injection of Imalgene and Rompun (7.5% / 2.5%; 0.1 mL / 10 g). Granulocyte infiltration and bone remodeling are measured using in vivo molecular imaging (Bruker In-Vivo Xtreme imaging system; excitation wavelength:

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630 nm, emission wavelength: 700 nm, acquisition time: 5 seconds for the ProSense® 680 probe; excitation wavelength: 720 nm, emission wavelength: 790 nm, acquisition time: 5 seconds for the OsteoSense®750EX probe).

[538] There are 10 mice per group. The results are expressed as mean ± SEM and statistical analysis is performed using unidirectional ANOVA, followed by Dunnett's post-hoc test versus vehicle / patient group for score analysis and imaging versus vehicle / simulated body weight group.

6.8. CIA model

6.8.1. Materials

[539] Freund's complete adjuvant (CFA) and Freund's incomplete adjuvant (IFA) were purchased from Difco. Bovine type II collagen (Cll), lipopolysaccharide (LPS) and Enbrel were obtained from Chondrex (Isle d'Abeau, France); Sigma (P4252, Lisle d'Abeau, France), Whyett (25 mg injectable syringe, France) Acros Organics (Paio Alto, CA), respectively. All other reagents used were reagent grade and all solvents were analytical grade.

6.8.2. Animals

[540] Dark Agouti rats (male, 7-8 weeks old) were obtained from Harlan Laboratories (Maison-Alfort, France). The rats were maintained on a 12-hour light / dark cycle (07: 0019: 00). The temperature was maintained at 22 ° C and food and water were provided ad libitum.

6.8.3. Collagen-induced arthritis (ASD)

[541] One day before the experiment, CLL solution (2 mg / ml) was prepared with 0.05 M acetic acid and stored at 4 ^Q C. Immediately before immunization, equal volumes of adjuvant (IFA) were Cll and mixed by a homogenizer in a pre-cooled glass bottle in an ice water bath. It may be necessary

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143/154 extra adjuvant and extended homogenization if an emulsion is not formed. 0.2 ml of the emulsion was injected intradermally at the base of each rat's tail on day 1, a second intradermal booster injection (2 mg / ml Cll solution in 0.1 ml saline with CFA) on day 9. This method of immunization has been modified from published methods (Sims etal., 2004; Jou etal., 2005).

6.8.4. Study design

[542] The therapeutic effects of the compounds were tested in the CIA model in rats. The rats were randomly divided into equal groups and each group contained 10 rats. All rats were immunized on day 1 and boosted on day 9. Therapeutic dosage lasted from day 16 to day 30. The negative control group was treated with vehicle (0.5% MC) and the positive control group with Enbrel (10 mg / kg, 3x week, sc). A compound of interest was typically tested at 4 doses, for example, 1.3, 10, 30 mg / kg.

6.8.5. Clinical evaluation of arthritis

[543] Arthritis is scored according to the method of Khachigian, 2006, Lin etal., 2007 and Nishida etal., 2004. The swelling of each of the four legs is classified with the arthritic score as follows: 0 - without symptoms; 1 - mild, but defined redness and swelling of a type of joint, such as ankle or wrist or apparent redness and swelling limited to individual fingers, regardless of the number of fingers affected; 2 - moderate redness and swelling of two or more types of joints; 3 - severe redness and swelling of the entire paw, including fingers; 4 - extremely inflamed limb with multiple joint involvement (maximum cumulative score for clinical arthritis of 16 per animal) (Nishida et al., 2004).

[544] To allow for meta-analysis of multiple studies, clinical score values have been normalized as follows:

[545] ® AUC of clinical score (AUC score): area under

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144/154 the curve (AUG) from day 1 to day 14 was calculated for each individual rat. The AUG of each animal was divided by the average AUG obtained for the vehicle in the study, from which data on this animal were obtained and multiplied by 100 (that is, the AUG was expressed as a percentage of the average AUG of the vehicle per study ).

[546] ® Increase in clinical score from day 1 to day 14 (final score): The difference in clinical score for each animal was divided by the average difference in clinical score obtained for the vehicle in the study, from which data on this animals were obtained and multiplied by 100 (ie, the difference was expressed as a percentage of the average difference in clinical score for the vehicle per study).

6.8.5.1. Variation in body weight (%) after the onset of arthritis

[547] Clinically, weight loss is associated with arthritis (Shelton et al., 2005; Argiles et al., 1998; Rali, 2004; Walsmith et al., 2004). Thus, changes in body weight after the onset of arthritis can be used as a non-specific parameter to assess the effect of the therapeutic product on the rat model. The variation in body weight (%) after the onset of arthritis was calculated as follows:

Weight ccrsorai - Weight corswrai 6}

CamtiBdoBgos; .............. ^: ................................... .........'........................ x J 00%

Body weight fsemans 5}

Body weight (sssane 41 - Body weight 3>

Rsígs: ................................................ ..................................... xlD0%

Colpcrat weight (mountain range)

6.8.5.2. Radiology

[548] Radiographs of the hind legs of each individual animal were taken. A random identity number was assigned blindly for each of the radiographs and the severity of bone erosion was classified by two independent markers with the Larsen radiological scoring system as follows: 0 - normal with intact bone outlines and normal joint space; 1 - mild abnormality with any one or two of the external metatarsal bones present

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145/154 with slight bone erosion; 2 - premature abnormality defined with three to five of the external metatarsal bones showing bone erosion; 3 - median destructive abnormality with all external metatarsal bones, as well as any one or two of the internal metatarsal bones that present defined bone erosions; 4 - severe destructive abnormality with all metatarsal bones showing definite bone erosion and at least one of the internal metatarsal joints completely eroded, leaving some contours of the bone joint partially preserved; 5 - mutilating abnormality without bone contours. This scoring system is a modification by Salvemini et al., 2001; Bush etal., 2002; Sims etal., 2004; Jou etal., 2005.

6.8.5.3. Histology

[549] After radiological analysis, the mice's hind legs were fixed in 10% formalin buffered with phosphate (pH 7.4), decalcified with rapid bone decalcifier for fine histology (Laboratories Eurobio) and embedded in paraffin. To ensure an extensive assessment of arthritic joints, at least four sections in series (5 μιτι thick) were cut and each series of sections had 100 μιτι between them. The sections were stained with hematoxylin and eosin (H&E). Histological exams for synovial inflammation and bone and cartilage damage were performed in double blind. In each leg, four parameters were assessed on a four-point scale. The parameters were cell infiltration, pannus severity, cartilage erosion and bone erosion. Scoring was performed according to the following: 1 - normal, 2 - mild, 3 - moderate, 4 accentuated. These four scores are added together and represented as an additional score, namely the total RA score.

6.8.5.4. Computed tomography (pCT) analysis of the calcaneus (heel bone):

[550] Bone degradation seen in RA occurs especially

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146/154 in the cortical bone and can be revealed by pCT analysis (Sims NA et al, Arthritis Rheum. 50 (2004) 2338-2346: Targeting Osteoclasts With Zoledronic Acid Prevents Bone Destruction in CollagenInduced Arthritis; Oste L. et al., ECTC Montreal 2007: A High Throughput Method of Measuring Bone Architectural Disturbance in a Murine CIA Model by Micro-CT Morfometry). After scanning and 3D reconstruction of the calcaneus bone volume, bone degradation is measured as the number of distinct objects present per section, isolated in silico perpendicular to the longitudinal axis of the bone. The more the bone is degraded, the more distinct objects are measured. 1000 sections are analyzed evenly distributed along the calcaneus (spaced at about 10.8 pm).

6.8.5.5. Steady state PK

[551] On days 7 or 11, blood samples were collected from the retro-orbital sinus with lithium heparin as an anticoagulant at the following times: pre-dose, 1, 3 and 6 hours. The whole blood samples were centrifuged and the resulting plasma samples were stored at -20 ° C, awaiting analysis. The plasma concentrations of each test compound were determined using an LC-MS / MS method, in which the mass spectrometer was operated in the positive electrospray mode. Pharmacokinetic parameters were calculated using Winnonlin (Pharsight, United States) and assuming that pre-dose plasma levels were equal to plasma levels within 24 hours.

6.8.6. Results

Table XI. Clinical score

Day CIA vehicle Cpd 1 (1 mg / kg) Cpd 1 (3 mg / kg) Cpd 1 (10 mg / kg) Cpd 1 (30 mg / kg) 31 2.3 0.448 2.3 2.2 2.2 2.3 32 3 0.537 2.6 2.6 2.5 2 33 3.7 0.578 3.1 2.7 2.4 2 34 4.6 0.562 3.7 2.8 2 1.7

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35 5.5 0.637 4.5 3 2 1.8 38 7.5 0.86 6.5 4.1 4.1 3.9 39 7.9 0.849 7.5 4.9 4.8 4.1 40 7.8 0.827 7.6 5.3 6.4 4.4 41 8.2 0.964 7.4 5.3 6.4 5.4 42 7.9 0.948 7.6 5.6 6.6 5.9 45 7.9 0.752 8.4 7.1 6.7 7 46 7.8 0.786 8.2 7.3 6.3 7.1

Table XII. Clinical score (p-values)

Day Vehicle Cpd XXb (7.5 mg / kg bid) Cpd XXb (15 mg / kg bid) Cpd 1 + Cpd XXb (30+ 15 mg / kg) 31 2.9 (1) 3 (0.889) 3 (0.889) 3 (0.889) 32 3.5 (1) 3.6 (0.8664) 3.8 (0.6661) 3.2 (0.1964) 34 4.1 (1) 3.8 (0.6338) 3.9 (0.7869) 2.5 (0.1387) 35 4.4 (1) 3.7 (0.3327) 3.9 (0.5456) 2 (0.2409) 38 5.8 (1) 4.6 (0.2354) 4.2 (0.1094) 1.9 (0.0003) 39 7.1 (1) 4.7 (0.0712) 4.5 (0.0467) 2.2 (0.0001) 40 8 (1) 5.1 (0.038) 4.6 (0.0152) 2.6 (0.0267) 41 8.1 (1) 4.7 (0.0185) 4.3 (0.0085) 2.3 (0.0002) 42 8.3 (1) 4.7 (0.012) 4.6 (0.0104) 2.1 (0.0001) 45 10.1 (1) 5.3 (0.0024) 6.1 (0.0080) 2.9 (0.0001)

6.8.7. Conclusions

[552] As can be seen above, when taken together, the combination of the IRAK inhibitor and JAK results in a better effect than for each compound taken individually.

[553] This, in turn, can allow you to achieve the same or a better therapeutic effect with a smaller amount of drug. This can be particularly beneficial to avoid taking unnecessary amounts of the drug, while maintaining effectiveness and thus reducing the risk of drug-induced side effects.

FINAL CONSIDERATIONS

[554] It will be appreciated by those skilled in the art that the preceding descriptions are of an exemplary and explanatory nature, and are intended to illustrate the invention and its preferred modalities. Through routine experimentation, those skilled in the art will recognize obvious changes and variations that can be

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148/154 made without departing from the spirit of the invention. All such modifications that fall within the scope of the appended claims must be included in it. Thus, the invention is intended to be defined not by the above description, but by the following claims and their equivalents.

[555] All publications including, but not limited to, patents and patent applications, cited in this specification are hereby incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated herein by reference as if fully presented.

[556] It should be understood that factors such as the differential cell penetration capacity of the various compounds can contribute to the discrepancies between the activity of the compounds in biochemical and cellular in vitro assays.

[557] At least some of the chemical names of the compound of the invention as provided and presented in the present application may have been generated on an automated basis by using a commercially available chemical nomenclature software program, and have not been independently verified. Representative programs that perform this function include the Lexichem naming tool sold by OpenEye Scientific Software, Inc. and the Autonom Software tool sold by MDL Information Systems, Inc. In the event that the indicated chemical name and the structure described differ, the structure represented will prevail.

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Claims (17)

1. Composition characterized by the fact of understanding: a) a compound according to Formula I

CN where: Cy is - C3-7 monocyclic cycloalkyl optionally substituted by one or more independently selected R ³ , or - 4- to 7-element heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more independently selected R ³ ; R ¹ is - H, --SO3H, - -P (= O) (OH) 2, - C1-4 alkyl, - -C (= O) - (monocyclic heterocycloalkyl of 4 to 7 elements comprising one or two heteroatoms independently selected from N, S and O) or - -C (= O) C1-6 alkyl, C1-6 alkyl which is optionally substituted by one or more independently selected R ⁴ groups; R ² is H or C 1-4 alkyl; each R ³ is independently selected from: -Oh, Petition 870190079079, of 8/15/2019, p. 177/183 2/5 - = 0, - halo, and - C1-4 alkyl; each R ⁴ is independently selected from: - -NR ^5a R ^5b , - -C (= O) OH, - 4- to 7-element monocyclic heterocycloalkyl comprising one or two heteroatoms independently selected from N, S and O, optionally substituted by one or more C1-4 alkyl independently selected and - -NHC (= 0) -Ci-4 alkyl-NH ₂ ; and R ^5a and R ^5b are independently H or C1-4 alkyl; or a salt or solvate or a pharmaceutically acceptable solvate salt thereof; and b) a second compound that has a JAK inhibitory activity. 2. Composition according to claim 1, characterized by the fact that the compound or pharmaceutically acceptable salt thereof according to Formula I, Cy is monocyclic C 3-7 cycloalkyl substituted by one or two independently selected from R ^3. Composition according to claim 1 or 2, characterized in that, in the compound or pharmaceutically acceptable salt thereof according to Formula I, Cy is tetrahydropyranyl or tetrahydrothiopyranyl, each of which is optionally substituted by one or two R ³ is independently selected. Composition according to any one of claims 1 to 3, characterized in that, in the compound or pharmaceutically acceptable salt thereof according to Formula I, R ³ is selected from OH, = 0, F, and -CH3. Petition 870190079079, of 8/15/2019, p. 178/183 3/5 5. Composition according to claim 1, characterized by the fact that the compound or pharmaceutically acceptable salt thereof according to Formula I is according to Formula Ha, lib, He, lld, He or Hf:

Composition according to any one of claims 1 to 5, characterized in that, in the compound or pharmaceutically acceptable salt thereof according to any one of Formulas l-llf, R ¹ is H, -CH ₃ , - SO ₃ H or -P (= O) (OH) ₂ . Composition according to any one of claims 1 to 5, characterized in that, in the compound or pharmaceutically acceptable salt thereof according to any one of Formulas l-llf, R ¹ is -C (= O) Ci -6 alkyl, C1-6 alkyl which is replaced by one or two independently selected R ⁴ . Composition according to any one of claims 1 to 5, characterized in that, in the compound or pharmaceutically acceptable salt thereof according to any one of Formulas l-llf, R ¹ is -C (= O) Ci -6 alkyl, C1-6 alkyl which is replaced by one or two independently selected -C (= O) OH, -NH ₂ , -NHCH ₃ or -N (CH ₃ ) ₂ . 9. Composition according to any one of claims 1 to 8, characterized by the fact that, in the compound, the salt or Petition 870190079079, of 8/15/2019, p. 179/183 4/5 pharmaceutically acceptable thereof according to any of Formulas l-llf, R ² is H or CH3. 10. Composition according to claim 1, characterized by the fact that the compound according to Formula I is selected from: 6- [6- [2- (2-hydroxy-ethoxy) -ethoxy] -5- (tetrahydro-pyran-4-ylamino) imidazo [4,5-b] pyridin-3-yl] -nicotinonitrile, and 2- {2- [3- (5-cyano-pyridin-2-yl) -5- (tetrahydro-pyran-4ylamino) -3H-imidazo [4,5-b] pyridin-6-yloxy] - ester (S) -2amino-3-methyl-butyric acid ethoxy} -ethyl. Composition according to any one of claims 1 to 10, characterized in that the JAK inhibitor compound is a JAK1 inhibitor. 12. Composition according to any one of claims 1 to 10, characterized in that the JAK I inhibiting compound is according to Formula XXa or XXb:

XXu XXb Pharmaceutical composition characterized by the fact that it comprises a pharmaceutically effective amount of a composition as defined in any one of claims 1 to 12 and a pharmaceutically acceptable carrier. 14. Pharmaceutical composition according to claim 13, characterized in that it comprises another therapeutic agent. Petition 870190079079, of 8/15/2019, p. 180/183 5/5 15. Pharmaceutical composition, according to claim 14, characterized by the fact that the other therapeutic agent is an agent for the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases, transplant rejection, diseases that involve impairment of cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons. 16. Composition according to any one of claims 1 to 12 or pharmaceutical composition according to any one of claims 13 to 15, characterized in that they are for use in medicine. Composition according to any one of claims 1 to 12 or pharmaceutical composition according to any one of claims 13 to 15, characterized in that they are for use in the prophylaxis and / or treatment of inflammatory diseases, autoimmune diseases, proliferative diseases, allergic diseases , transplant rejection, diseases that involve impaired cartilage renewal, congenital cartilage malformations and / or diseases associated with hypersecretion of IL6 or interferons.