Methods for blocking or ameliorating cytokine release syndrome

Abstract

Disclosed herein are embodiments of methods for treating or preventing cytokine release syndrome (CRS). In certain embodiments, the methods include administering a compound, or a salt, solvate, prodrug, or pharmaceutical composition thereof, to a subject suffering from or at risk of developing CRS. The compound may be a Syk inhibitor and / or may have a structure represented by Formula I. The methods may also include administering the compound to a subject who has undergone, is currently undergoing, and / or will undergo cell therapy. TIFF2022544276000034.tif28128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit under 35 U.S.C. §119(e) of the filing date of earlier-filed U.S. Provisional Patent Application No. 62 / 886,806, filed August 14, 2019, the entirety of which is incorporated herein by reference.

[0002] Field The present application relates to compounds, as well as salts, solvates and / or prodrugs thereof, and pharmaceutical compositions containing them, and methods of using the compounds, salts, solvates, prodrugs and / or compositions to treat cytokine release syndrome. [Background technology]

[0003] background Cytokine release syndrome (CRS) is a potentially life-threatening disease that can result from a variety of factors, including severe viral infections such as influenza, antibodies used in immunotherapies such as cancer immunotherapy, and administration of non-protein-based anticancer drugs such as oxaliplatin and lenalidomide. Immunotherapies can involve high levels of immune activation that exceed naturally occurring levels, and CRS is a non-antigen-specific toxicity that can result. As immune-based therapies become more potent, CRS is becoming increasingly diagnosed. CRS has also been observed in the setting of stem cell transplants from haploidentical donors and graft-versus-host disease. Shimabukuro-Vornhagen et al., Journal for ImmunoTherapy of Cancer 6:56 (2018). CRS has been associated with elevated blood levels of several cytokines, including interleukin (IL)-6 and interferon-gamma. Lee et al., Blood 124(2):188-195 (10 July 2014; Epub 29 May 2014) (Non-patent document 2).

[0004] CRS is typically observed clinically when a significant number of lymphocytes and / or myeloid cells are activated and release inflammatory cytokines. Cytokine release can be induced by chemotherapy or biological therapy and / or associated with treatment with therapeutic antibodies, such as immunotherapy for cancer treatment. Exemplary immunotherapies that can cause CRS include, but are not limited to, therapy in which cells express recombinant receptors, such as chimeric antigen receptors (CARs) and / or other transgenic receptors, such as T cell receptors (TCRs). CRS induced by CAR T therapy generally occurs within a few days of T cell infusion at the peak of CAR T cell proliferation. Giavridis et al., Nat Med. 24(6):731-738 (June 2018; Epub 28 May 2018) (Non-Patent Document 3). Examples of CAR T therapies that can induce CRS include axicabtagenecilloreucel (sold as YESCARTA®) and tisagenlecleucel (sold as KYMRIAH®).

[0005] Highly elevated interleukin 6 (IL-6) levels have been observed in patients with CRS and even in mouse models of the disease, indicating that IL-6 may play a role in the pathophysiology of CRS. Shimabukuro-Vornhagen, J Immunother Cancer 6(1), 56(2018). IL-6 can signal by two different modes. Classical IL-6 signaling involves the binding of IL-6 to the membrane-bound IL-6 receptor. However, the IL-6 receptor does not have an intracellular signaling domain. Instead, after soluble IL-6 binds to the membrane-bound IL-6 receptor, the IL-6 / IL-6 receptor complex binds to membrane-bound gp130, which initiates signaling through its intracellular domain. In trans-signaling, IL-6 binds to a soluble form of the IL-6 receptor, which is typically cleaved from the cell surface by metalloproteinases. The resulting soluble IL-6 / IL-6 receptor complex binds to gp130 and is therefore capable of inducing signaling even in cell types that do not express membrane-bound IL-6 receptors.

[0006] IL-6 contributes to many of the major symptoms of CRS. Through trans-signaling, IL-6 leads to the hallmark symptoms of severe CRS, namely vascular leakage and the coagulation cascade that induces complement activation and disseminated intravascular coagulation (DIC). In addition, IL-6 likely contributes to the cardiomyopathy often observed in patients with CRS by promoting myocardial dysfunction. In mouse models, CRS develops within 2-3 days of CAR T cell infusion and can be fatal. Giavridis et al., Nat Med. 24(6):731-738(2018) (Non-Patent Document 3). CRS symptoms can begin within minutes or hours of initiation of antibody treatment and can include fever that can reach or exceed 40°C, nausea, fatigue, headache, tachycardia, hypotension, rash, shortness of breath, and / or muscle pain. However, in some cases, additional, potentially more serious complications may occur, including cardiac dysfunction, adult respiratory distress syndrome, neurotoxicity, renal and / or hepatic failure, and / or disseminated intravascular coagulation.

[0007] The National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE v.5.0, pub. November 27, 2017) (Non-Patent Document 4) includes a scoring system for CRS. Grade 1: Fever with or without systemic symptoms. Grade 2: Hypotension in response to fluids; hypoxia in response to <40% O2. Grade 3: Hypotension controlled by one vasopressor; hypoxia requiring ≥40% O2. Grade 4: Life-threatening consequences; urgent intervention is indicated. Grade 5: Death. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] Shimabukuro-Vornhagen et al., Journal for ImmunoTherapy of Cancer 6:56(2018) [Non-Patent Document 2] Lee et al.,Blood 124(2):188-195(10 July 2014;Epub 29 May 2014) [Non-Patent Document 3] Giavridis et al., Nat Med.24(6):731-738(June 2018;Epub 28 May 2018) [Non-Patent Document 4] The National Cancer Institute Common Terminology Criteria for Adverse Events(CTCAE v.5.0,pub.November 27,2017) Summary of the Invention

[0009] overview Disclosed herein are embodiments of methods for treating or preventing CRS. In some embodiments, the methods include administering an effective amount of a compound to a subject suffering from or at risk of developing CRS. The compound can be a kinase modulator and / or inhibitor, such as a spleen tyrosine kinase (Syk) modulator and / or inhibitor. The compound can be a pyrimidinediamine compound and / or have a structure represented by Formula I, or a salt, solvate, N-oxide, and / or prodrug thereof. TIFF0007681570000001.tif28128

[0010] With respect to formula I, Y is CH 2 , N.R. 24 , O, S, S(O) and S(O) 2 Selected from Z 1 and Z 2 are each independently selected from CH and N. 2 are the same or different R 8lower alkyl, optionally substituted with one or more groups, the same or different R 8 lower cycloalkyl, optionally substituted with one or more groups, the same or different R 8 cyclohexyl, which may be substituted with one or more of the following groups: 8 3-8 membered heterocycloalkyl optionally substituted with one or more groups, the same or different R 8 may be substituted with one or more of the following groups (C 6 ~C 14 ) aryl, the same or different R 8 Phenyl, optionally substituted with one or more groups, and the same or different R 8 R is selected from the group consisting of 5- to 15-membered heteroaryl, optionally substituted with one or more groups. 5 is selected from halo, cyano, nitro, or trihalomethyl. 8 are independently a , R b , the same or different R a or R b R replaced by one or more of a , the same or different R a or R b -OR replaced by one or more of a , -B(OR a ) 2 , -B(NR c R c ) 2 , -(CH 2 ) m -R b , -(CHR a ) m -R b , -O-(CH 2 ) m -R b , -S-(CH 2 ) m -R b , -O-CHR a R b , -O-CR a (R b ) 2 , -O-(CHR a ) m -R b , -O-(CH 2) m -CH[(CH 2 ) m R b R b 、 -S-(CHR a ) m -R b 、 -C(O)NH-(CH 2 ) m -R b 、 -C(O)NH-(CHR a ) m -R b 、 -O-(CH 2 ) m -C(O)NH-(CH 2 ) m -R b 、 -S-(CH 2 ) m -C(O)NH-(CH 2 ) m -R b 、 -O-(CHR a ) m -C(O)NH-(CHR a ) m -R b 、 -S-(CHR a ) m -C(O)NH-(CHR a ) m -R b 、 -NH-(CH 2 ) m -R b 、 -NH-(CHR a ) m -R b 、 -NH[(CH 2 ) m R b 、 -N[(CH 2 ) m R b 2 、 -NH-C(O)-NH-(CH 2 ) m -R b 、 -NH-C(O)-(CH 2 ) m -CHR b R b 、及び -NH-(CH 2 ) m -C(O)-NH-(CH 2 )​m -R b is selected from.

[0011] R 17 is selected from hydrogen, halogen, or lower alkyl; R 18 is selected from hydrogen, halogen, lower alkyl, or alternatively, R 18 is R 17 R may be taken together with the carbon atom to which they are attached to form an oxo (=O) group, or may be taken together with the carbon atom to form a spiro ring containing 3 to 7 carbon atoms. 19 is selected from hydrogen or lower alkyl; R 20 is selected from hydrogen, or lower alkyl, or alternatively, R 20 is R 19 may be taken together with the carbon atom to which they are attached to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms.

[0012] R 21 , R 22 and R 23 Each R is independently selected from hydrogen or phosphonooxyalkyl. 24 is selected from hydrogen, lower alkyl, or phosphonooxyalkyl. 21 , R 22 , R 23 and R 24 At least one, for example one, two, three, or four, of R is phosphonooxyalkyl. 21 , R 22 , R 23 and R 24 is phosphonooxyalkyl, and in certain embodiments, R 21 is phosphonooxyalkyl. In some embodiments, R 21 , R 22 , R 23 and R 24 At least one, for example 1, 2, 3, or 4, of R is hydrogen. 21 , R 22 , R23 and R 24 are hydrogen, and in certain embodiments, R 21 , R 22 , R 23 and R 24 are all hydrogen. In another embodiment, R 21 is phosphonooxyalkyl, R 22 , R 23 and R 24 is hydrogen.

[0013] Each R a are each independently hydrogen, lower alkyl, lower cycloalkyl, cyclohexyl, (C 4 ~C 11 ) cycloalkylalkyl, (C 6 ~C 10 )Aryl, phenyl, (C 7 ~C 16 ) arylalkyl, benzyl, 2- to 6-membered heteroalkyl, 3- to 8-membered heterocycloalkyl, 4- to 11-membered heterocycloalkylalkyl, 5- to 10-membered heteroaryl, and 6- to 16-membered heteroarylalkyl. Each R b are, independently, =O, -OR a , (C 1 ~C 3 )Haloalkyloxy, =S, -SR a , =NR a , =NOR a , -NR c R c , halogen, -CF 3 , -CN, -NC, -OCN, -SCN, -NO, -NO 2 , =N 2 , -N 3 , -S(O)R a , -S(O) 2 R a , -S(O) 2 OR a , -S(O)NR c R c , -S(O) 2 NR c R c , -OS(O)R a , -OS(O) 2 Ra , -OS(O) 2 OR a , -OS(O) 2 NR c R c , -C(O)R a , -C(O)OR a , -C(O)NR c R c , -C(NH)NR c R c , -C(NR a )NR c R c , -C(NOH)R a , -C(NOH)NR c R c , -OC(O)R a , -OC(O)OR a , -OC(O)NR c R c , -OC(NH)NR c R c , -OC(NR a )NR c R c , -[NHC(O)] n R a , -[NR a C(O)] n R a , -[NHC(O)] n OR a , -[NR a C(O)] n OR a , -[NHC(O)] n NR c R c , -[NR a C(O)] n NR c R c , -[NHC(NH)] n NR c R c or -[NR a C(NR a )] n NR c R c Each R c are, independently of each other, R aor an amino protecting group selected from formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl, tert-butoxycarbonyl, trimethylsilyl, 2-trimethylsilyl-ethanesulfonyl, trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, or nitro-veratryloxycarbonyl, or alternatively, two R c may, together with its nitrogen atom, contain one or more of the same or different additional heteroatoms and may be the same or different R a It forms a 5-8 membered heterocycloalkyl or heteroaryl, which may be substituted with one or more groups.

[0014] Each m is, independently of the other, an integer from 1 to 3; and / or each n is, independently of the other, an integer from 0 to 3.

[0015] In some embodiments, the compound is TIFF0007681570000002.tif53147, or a salt, solvate, N-oxide, or prodrug thereof, wherein R 30 is H or phosphonooxyalkyl. In certain embodiments, the compound is TIFF0007681570000003.tif36140 or a salt and / or solvate thereof, for example The file is TIFF0007681570000004.tif35128.

[0016] In any of the methods, the subject may not exhibit signs or symptoms of CRS and / or may be at risk for developing CRS, in such embodiments, administering the compound substantially prevents the onset of CRS or prevents the onset of Grade 2 or higher CRS.

[0017] In other embodiments, the subject exhibits at least one sign or symptom of CRS, and may exhibit at least one sign or symptom of grade 1 CRS. Alternatively, the subject may exhibit at least one sign or symptom of grade 2 or higher CRS, such as grade 3 or higher CRS. The compound may be administered within 24 hours of onset of the sign or symptom, and / or administering the compound may improve the sign or symptom of CRS compared to the severity of the sign or symptom prior to administration of the compound, such as reducing the grade of CRS from 4 to 3, 2, or 1, or from 3 to 2, or 1, or from 2 to 1. Alternatively, the symptom of CRS is substantially reduced below grade 1 level, such that the subject no longer experiences symptoms associated with CRS. In some embodiments, the sign or symptom is fever, and may be a fever of 40° C. or higher.

[0018] The method may include administering to a subject who has previously received a first treatment for which CRS is a known, possible, or potential side effect. Administration of the first treatment may begin more than 0 to 10 days prior to administration of the compound. Alternatively, the compound may be administered to a subject who will be receiving or is currently receiving a first treatment for which CRS is a known, possible, and / or potential side effect. In any embodiment, the first treatment may include cell therapy, including but not limited to chimeric antigen receptor (CAR) expression therapy and / or transgenic receptor therapy. It is also known that acellular antibodies, including but not limited to CAMPATH 1-H, blinatumomab, and / or rituximab, can cause this syndrome, particularly those that activate T cells.

[0019] In some embodiments, the method may further include administering a second therapeutic agent, such as a steroid, an anti-inflammatory agent, an immunosuppressant, or a combination thereof. The steroid may be a corticosteroid, such as dexamethasone or prednisone, or a combination thereof. In any embodiment, the compound may be administered substantially simultaneously with the second therapeutic agent, or the compound and the second therapeutic agent may be administered sequentially in any order.

[0020] [The present invention 1001] A method for treating and / or preventing cytokine release syndrome (CRS), comprising administering to a subject suffering from or at risk of developing CRS an effective amount of a compound of formula I. TIFF0007681570000005.tif28128 (In the formula, Y is CH 2 , N.R. 24 , O, S, S(O) and S(O) 2 Selected from; Z 1 and Z 2 are each independently selected from CH and N; R 2 are the same or different R 8 lower alkyl, optionally substituted with one or more groups, the same or different R 8 lower cycloalkyl, optionally substituted with one or more groups, the same or different R 8 cyclohexyl, which may be substituted with one or more of the following groups: 8 3-8 membered heterocycloalkyl optionally substituted with one or more groups, the same or different R 8 may be substituted with one or more of the following groups (C 6 ~C 14 ) aryl, the same or different R 8 Phenyl, optionally substituted with one or more groups, and the same or different R 8 5 to 15 membered heteroaryl optionally substituted with one or more groups; R 5 is selected from halo, cyano, nitro, or trihalomethyl; Each R 8 are independently a 、R b , the same or different R a or R b R replaced by one or more of a , the same or different R a or R b -OR replaced by one or more of a , -B(OR a ) 2 , -B(NR c R c) 2 , -(CH 2 ) m -R b , -(CHR a ) m -R b , -O-(CH 2 ) m -R b , -S-(CH 2 ) m -R b , -O-CHR a R b , -O-CR a (R b ) 2 , -O-(CHR a ) m -R b , -O-(CH 2 ) m -CH[(CH 2 ) m R b ]R b , -S-(CHR a ) m -R b , -C(O)NH-(CH 2 ) m -R b , -C(O)NH-(CHR a ) m -R b , -O-(CH 2 ) m -C(O)NH-(CH2 ) m -R b , -S-(CH 2 ) m -C(O)NH-(CH 2 ) m -R b , -O-(CHR a ) m -C(O)NH-(CHR a ) m -R b , -S-(CHR a ) m -C(O)NH-(CHR a ) m -R b , -NH-(CH 2 ) m -R b , -NH-(CHR a ) m -R b , -NH[(CH 2 ) m R b ], -N[(CH 2 ) m R b ] 2 , -NH-C(O)-NH-(CH 2 ) m -R b , -NH-C(O)-(CH 2 ) m -CHR b R b , and -NH-(CH 2) m -C(O)-NH-(CH 2 ) m -R b Selected from; R 17 is selected from hydrogen, halogen, or lower alkyl; R 18 is selected from hydrogen, halogen, and lower alkyl; Or, alternatively, R 18 is R 17 may be taken together with the carbon atom to which they are attached to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms; R 19 is selected from hydrogen, or lower alkyl; R 20 is selected from hydrogen, or lower alkyl; Or, alternatively, R 20 is R 19 may be taken together with the carbon atom to which they are attached to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms; Each R a are each independently hydrogen, lower alkyl, lower cycloalkyl, cyclohexyl, (C 4 ~C 11 ) cycloalkylalkyl, (C 6 ~C 10 )Aryl, phenyl, (C 7 ~C 16 ) selected from arylalkyl, benzyl, 2- to 6-membered heteroalkyl, 3- to 8-membered heterocycloalkyl, 4- to 11-membered heterocycloalkylalkyl, 5- to 10-membered heteroaryl, and 6- to 16-membered heteroarylalkyl; Each R b are, independently, =O, -OR a 、(C 1 ~C 3 )Haloalkyloxy, =S, -SR a , =NR a , =NOR a , -NR c R c , halogen, -CF 3 , -CN, -NC, -OCN, -SCN, -NO, -NO 2 、=N2 、-N 3 , -S(O)R a , -S(O) 2 R a , -S(O) 2 OR a , -S(O)NR c R c , -S(O) 2 NR c R c , -OS(O)R a , -OS(O) 2 R a , -OS(O) 2 OR a , -OS(O) 2 NR c R c , -C(O)R a , -C(O)OR a , -C(O)NR c R c , -C(NH)NR c R c , -C(NR a )NR c R c , -C(NOH)R a , -C(NOH)NR c R c , -OC(O)R a , -OC(O)OR a , -OC(O)NR c R c , -OC(NH)NR c R c , -OC(NR a )NR c R c , -[NHC(O)] n R a , -[NR a C(O)] n R a , -[NHC(O)] n OR a , -[NR a C(O)] n OR a , -[NHC(O)] n NR c R c , -[NR a C(O)] n NR c R c , -[NHC(NH)] n NR c R c or -[NR a C(NR a )] n NR c R c Selected from; Each R c are, independently of each other, R a or an amino protecting group selected from formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl, tert-butoxycarbonyl, trimethylsilyl, 2-trimethylsilyl-ethanesulfonyl, trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl, or nitro-veratryloxycarbonyl; or alternatively, two R attached to the same nitrogen atom c may, together with its nitrogen atom, contain one or more of the same or different additional heteroatoms and may be the same or different R a forming a 5-8 membered heterocycloalkyl or heteroaryl, optionally substituted with one or more groups; R 21 、R 22 and R 23 are each independently selected from hydrogen or phosphonooxyalkyl; R24 is selected from hydrogen, lower alkyl, or phosphonooxyalkyl; Each m is, independently, an integer from 1 to 3; Each n is independently an integer from 0 to 3. or a salt, solvate, N-oxide, or prodrug thereof. [The present invention 1002] R 21 、R 22 、R 23 and R 24 The method of claim 1001, wherein at least one of is phosphonooxyalkyl. [The present invention 1003] R 21 、R 22 、R 23 and R 24 The method of the present invention 1001 or 1002, wherein at least one of is hydrogen. [The present invention 1004] R 21 is phosphonooxyalkyl, R 22 、R 23 and R 24 The method according to any one of claims 1001 to 1003, wherein is hydrogen. [The present invention 1005] The compound is TIFF0007681570000006.tif21128 or a salt, solvate, N-oxide, or prodrug thereof. [The present invention 1006] The compound is TIFF0007681570000007.tif24128 or a salt, solvate, N-oxide, or prodrug thereof. [The present invention 1007] The compound has the formula: TIFF0007681570000008.tif24128 or a salt, solvate, N-oxide, or prodrug thereof. [The present invention 1008] The compound has the formula: TIFF0007681570000009.tif23128 or a salt, solvate, N-oxide, or prodrug thereof, where R30 The method of any one of claims 1001 to 1007, wherein is H or phosphonooxyalkyl. [The present invention 1009] The compound is TIFF0007681570000010.tif35128 Or a salt and / or solvate thereof. [The present invention 1010] The compound is TIFF0007681570000011.tif35128 Any of the methods of the present invention 1001 to 1009, [The present invention 1011] The compound is TIFF0007681570000012.tif22128 The method according to any one of claims 1001 to 1008, [The present invention 1012] The method of any of claims 1001 to 1011, wherein administering the compound ameliorates a sign or symptom of CRS compared to the severity of the sign or symptom prior to administration of the compound. [The present invention 1013] The method of claim 1012, wherein said sign or symptom is fever. [The present invention 1014] The administering step comprises: administering to a subject who has previously received a first treatment for which CRS is a known, possible, or potential side effect; or Administered to a subject who is about to receive or is currently receiving a first treatment for which CRS is a known, possible, or potential side effect. Any of the methods of the present invention 1001 to 1013, comprising: [The present invention 1015] The method of claim 1014, wherein said first treatment comprises cell therapy. [The present invention 1016] The method of claim 1015, wherein the cell therapy comprises chimeric antigen receptor (CAR) expression therapy, transgenic receptor therapy, or a combination thereof. [The present invention 1017] The method of any of claims 1001 to 1016, wherein the step of administering the compound further comprises administering a second therapeutic agent. [The present invention 1018] The method of claim 1017, wherein said second therapeutic agent is a steroid, an anti-inflammatory agent, an immunosuppressant, or a combination thereof. [The present invention 1019] The steroid may be alclometasone, algestone, beclomethasone, betamethasone, budesonide, clobetasol, clobetasone, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, fludrocortisone, flumethasone, flunisolide, fluocinolone, fluocinonide, fluocortin, fluocortolone, fluorometh ... bromine, fluperolone, fluprednidene, fluprednisolone, flurandrenolide, fluticasone, formocortal, halcinonide, halobetasol, halometasone, halopredone, hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone, paramethasone, prednicarbate, prednisolone, prednisone, prednibal, prednylidene, rimexolone, tixocortol, triamcinolone, or any combination thereof; the anti-inflammatory agent is an aminosalicylate, a cyclooxygenase inhibitor, diclofenac, etodolac, famotidine, fenoprofen, flurbiprofen, ketoprofen, ketorolac, ibuprofen, indomethacin, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, salsalate, sulindac, tolmetin, or a combination thereof; or The method of the present invention, wherein the immunosuppressant is mercaptopurine, a corticosteroid, an alkylating agent, a calcineurin inhibitor, an inhibitor of inosine monophosphate dehydrogenase (IMPDH), an agent designed to preserve the recipient's humoral immune response intact while suppressing cellular immunity, or a combination thereof. [The present invention 1020] The method of claim 1018, wherein said second therapeutic agent is dexamethasone or prednisone, or a combination thereof. The above and other objects, features and advantages of the present technology will become more apparent from the following detailed description. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] Detailed Description I. Definition The following explanations of terms and methods are provided to more fully describe the present disclosure and to guide those skilled in the art in the practice of the present disclosure. The singular forms "a", "an" and "the" mean one or more, unless the context clearly dictates otherwise. The word "or" refers to a single element or a combination of two or more elements of the presented alternative elements, unless the context clearly dictates otherwise. As used herein, "comprises" means "includes." Thus, "comprises A or B" means "includes A, B or A and B" and does not exclude additional elements. All references cited herein, including patents and patent applications, are hereby incorporated by reference.

[0022] Unless otherwise specified, all numbers expressing amounts of ingredients, molecular weights, percentages, temperatures, times, etc. used in the specification or claims should be understood to be modified by the word "about". Thus, unless otherwise specified, either implicitly or explicitly, the numerical parameters recited are approximations that may depend on the desired properties sought and / or detection limits under standard testing conditions / methods. When directly and explicitly distinguishing an embodiment from the prior art presented, the numbers of that embodiment are not approximations unless the word "about" is recited.

[0023] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The materials, methods, and examples are for illustrative purposes only and are not intended to be limiting.

[0024] When chemical structures are depicted or described, unless expressly stated otherwise, all carbons are considered to include hydrogens such that each carbon has a valence of 4. For example, in the structure on the left in the schematic diagram below, the presence of 9 hydrogen atoms is implied. In the structure on the right, 9 hydrogen atoms are depicted. TIFF0007681570000013.tif22128

[0025] Identify specific atoms in the structure as hydrogen or containing hydrogen atoms with a string, e.g. -CH 2 CH 2 Those skilled in the art will appreciate that the notation described above is common in the chemical arts as a concise and simple way to represent organic structures.

[0026] The group R is not "fixed" to the ring system, for example the group shown below: When depicted as TIFF0007681570000014.tif14128, unless otherwise specified, the substituent R may be located on any atom of the fused bicyclic ring system so long as a stable structure is formed according to standard valence constraints understood by those of skill in the art. In the example shown, the R group may be located on an atom on either the 5- or 6-membered ring of the indolyl ring system, including heteroatoms by replacing hydrogens as explicitly stated, but the symbol TIFF0007681570000015.tif4128, excluding atoms bridging the carbon atoms.

[0027] When more than one such "free" group is depicted, for example in the formula: Where there are two groups, i.e., R and a bond, such as in TIFF0007681570000016.tif18128, which indicate a bond to a parent structure; unless otherwise specified, each "unfixed" group may be located on any atom on the ring system, which also contemplates that each of the depicted, implied, or explicitly defined hydrogens on the ring system may be replaced to form a chemically stable compound by any arrangement, etc.

[0028] Examples of R groups present on ring systems containing saturated carbons include those of the formula: When written as TIFF0007681570000017.tif11128, in this example, y can be greater than 1, assuming that each R replaces a hydrogen on the ring that is then depicted, implied, or explicitly defined; in that case, unless otherwise specified, two Rs can be located on the same carbon. A simple example is when R is a methyl group. In the illustrated structure, it can also exist as a geminal dimethyl on the illustrated ring carbon (an "annular" carbon). In another example, two Rs on the same carbon can form within a ring that includes that carbon, thus forming a spiro ring ("spirocyclic" group) structure. For example, the two Rs shown below are: A piperidine ring may be formed together with the cyclohexane ring in a spirocyclic arrangement such as TIFF0007681570000018.tif11128.

[0029] One of ordinary skill in the art will understand that definitions may be combined to further describe a particular compound, for example, hydroxyaliphatic refers to an aliphatic group substituted with a hydroxy (-OH) group, haloalkylaryl refers to an aryl group substituted with an alkyl group, where the alkyl group is also substituted with a halogen, and where aryl is the base name for the substituent, so that the point of attachment to the parent structure is through the aryl moiety.

[0030] As used herein, the term "substituted" refers to any modifier that follows a term, such as "substituted arylC 1~8 The term "alkyl" means "C 1~8 "Alkyl" moiety, "aryl" moiety or aryl C 1~8 Substitution can occur at both positions on the alkyl group. Also, by way of example, alkyl includes substituted cycloalkyl groups.

[0031] When "substituted" is used to modify a particular group or moiety, it means that at least one hydrogen atom, and possibly two or more hydrogen atoms, of that particular group or moiety are independently replaced with the same or different substituents as defined below. In certain embodiments, a group, moiety, or substituent may be substituted or unsubstituted unless specifically defined as either "unsubstituted" or "substituted". Thus, any of the groups defined herein may be substituted or unsubstituted. In certain embodiments, a substituent may or may not be specifically defined as substituted, but may still be considered to be substituted. For example, an "alkyl" or "pyrazolyl" moiety may be substituted or unsubstituted, but an "unsubstituted alkyl" or an "unsubstituted pyrazolyl" is not substituted.

[0032] A "substituent" or "substituent group" that replaces one or more hydrogen atoms on a saturated carbon atom of a particular group or moiety is represented by the formula -R, unless otherwise specified. 60 , halo, =O, -OR 70 , -SR 70 , -N(R 80 ) 2 , haloalkyl, perhaloalkyl, -CN, -NO 2 , =N 2 , -N 3 , -SO 2 R 70 , -SO 3 - M + , -SO 3 R 70 , -OSO 2 R 70 , -OSO 3 - M + , -OSO 3 R 70 , -P(O)(O - ) 2 (M + ) 2 , -P(O)(O - ) 2 M 2+ , -P(O)(OR70 )O - M + , -P(O)(OR 70 ) 2 , -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -CO 2 - M + , -CO 2 R 70 , -C(S)OR 70 , -C(O)N(R 80 ) 2 , -C(NR 70 )(R 80 ) 2 , -OC(O)R 70 , -OC(S)R 70 , -OCO 2 - M + , -OCO 2 R 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 CO 2 - M + , -NR 70 CO 2 R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)N(R 80 ) 2 , -NR 70 C(NR 70 )R 70 or -NR 70 C(NR 70 )N(R 80 ) 2 and R 60 is C 1~10 Aliphatic, heteroaliphatic or alicyclic, typically C 1~6 Aliphatic, more typically C 1~6 is alkyl, R 60 may be substituted; each R 70is independently at each occurrence hydrogen or R 60 and each R 80 independently for each occurrence, R 70 or alternatively, two R 80 The groups, together with the nitrogen atom to which they are attached, form a 3- to 7-membered aliphatic heterocycle which may contain 1 to 4 identical or different additional heteroatoms selected from O, N and S, of which N is R 70 Substituents such as H or C 1 ~C 3 Each M may have an alkyl substituent. + is a counterion with a net positive charge. + may independently represent, at each occurrence, an alkali metal ion, e.g., K + , Na + , or Li + ammonium ion, e.g. + N(R 70 ) 4 a protonated amino acid ion, such as a lysine ion or an arginine ion; or an alkaline earth metal ion, such as [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 , or [Ba 2+ ] 0.5 (The subscript "0.5" means, for example, that one of the counterions to such divalent alkaline earth metal ions is an ionized form of a compound of the invention and the other may be a typical counterion such as chloride, or that a two-ionized compound may serve as the counterion to such divalent alkaline earth ions, or alternatively, that a doubly ionized compound may serve as the counterion to such divalent alkaline earth ions.) As a specific example, -N(R 80 ) 2 is -NH 2 , -NH-alkyl, -NH-pyrrolidin-3-yl, N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl, N-morpholinyl, etc. Any two hydrogen atoms on a single carbon can be, for example, =O, =NR 70, =N-OR 70 , =N 2 Or it may be replaced by =S.

[0033] Substituents replacing hydrogen atoms on unsaturated carbon atoms of groups containing unsaturated carbons are -R unless otherwise specified. 60 , halo, -O - M + , -OR 70 , -SR 70 , -S - M + , -N(R 80 ) 2 , perhaloalkyl, -CN, -OCN, -SCN, -NO, -NO 2 , -N 3 , -SO 2 R 70 , -SO 3 - M + , -SO 3 R 70 , -OSO 2 R 70 , -OSO 3 - M + , -OSO 3 R 70 , -PO 3 -2 (M + ) 2 , -PO 3 -2 M 2+ , -P(O)(OR 70 )O - M + , -P(O)(OR 70 ) 2 , -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -CO 2 - M + , -CO 2 R 70 , -C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )N(R 80 ) 2, -OC(O)R 70 , -OC(S)R 70 , -OCO 2 - M + , -OCO 2 R 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 CO 2 - M + , -NR 70 CO 2 R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)N(R 80 ) 2 , -NR 70 C(NR 70 )R 70 and -NR 70 C(NR 70 )N(R 80 ) 2 and R 60 , R 70 , R 80 and M + is as defined above. In the case of an alkene or alkyne, the substituent is -O - M + , -OR 70 , -SR 70 Or -S - M + isn't it.

[0034] The substituent replacing the hydrogen atom on the nitrogen atom of such a nitrogen-containing group is, unless otherwise specified, -R 60 , -O - M + , -OR 70 , -SR 70 , -S - M + , -N(R 80 ) 2 , perhaloalkyl, -CN, -NO, -NO 2 , -S(O) 2 R 70, -SO 3 - M + , -SO 3 R 70 , -OS(O) 2 R 70 , -OSO 3 - M + , -OSO 3 R 70 , -PO 3 2- (M + ) 2 , -PO 3 2- M 2+ , -P(O)(OR 70 )O - M + , -P(O)(OR 70 )(OR 70 ), -C(O)R 70 , -C(S)R 70 , -C(NR 70 )R 70 , -CO 2 R 70 , -C(S)OR 70 , -C(O)NR 80 R 80 , -C(NR 70 )NR 80 R 80 , -OC(O)R 70 , -OC(S)R 70 , -OCO 2 R 70 , -OC(S)OR 70 , -NR 70 C(O)R 70 , -NR 70 C(S)R 70 , -NR 70 CO 2 R 70 , -NR 70 C(S)OR 70 , -NR 70 C(O)N(R 80 ) 2 , -NR 70 C(NR 70 )R 70 Or -NR 70 C(NR 70 )N(R 80 )2 and R 60 , R 70 , R 80 and M + is synonymous with the above.

[0035] In one embodiment, a substituted group has 1 substituent, 2 substituents, 3 substituents or 4 substituents.

[0036] Also, in embodiments where a group or moiety is substituted with substituted substituents, the nesting of such substituted substituents is limited to three, thereby preventing the formation of polymers. Thus, in a group or moiety that includes a first group that is a substituent on a second group (which is itself a substituent on a third group attached to the parent structure), the first (outermost) group can only be substituted with unsubstituted substituents. For example, in a group that includes -(aryl-1)-(aryl-2)-(aryl-3), aryl-3 can itself only be substituted with unsubstituted substituents.

[0037] "Aliphatic" refers to a group or moiety that is substantially hydrocarbon-based. An aliphatic group or moiety can be in its acyclic form, including alkyl, alkenyl, or alkynyl groups, its cyclic form (such as alicyclic groups or moieties, including cycloalkyl, cycloalkenyl, or cycloalkynyl), and further includes linear and branched configurations, and all stereo and positional isomers. Unless expressly stated otherwise, an aliphatic group contains from 1 to 25 carbon atoms (C 1~25 For example, in the case of a saturated acyclic aliphatic group or moiety, 1 to 15 (C 1~15 ), 1~10 pieces (C 1~10 ), 1~6 pieces (C 1~6 ), or 1 to 4 carbon atoms (C 1~4 ), 2 to 25 carbon atoms (C 2~25 For example, in the case of an unsaturated acyclic aliphatic group or moiety, 2 to 15 (C 2~15 ), 2~10 pieces (C 2~10 ), 2~6 pieces (C 2~6 ), or 2 to 4 carbon atoms (C 2~4 ), or in the case of an alicyclic group or moiety, 3 to 15 (C3~15 ), 3~10 pieces (C 3~10 ), 3~6 pieces (C 3~6 ), or 3 to 4 (C 3~4 ) carbon atoms. An aliphatic group may be substituted or unsubstituted, unless expressly designated as "unsubstituted aliphatic" or "substituted aliphatic." An aliphatic group may be substituted with one or more substituents (up to two substituents for each methylene carbon in the aliphatic chain, or up to one substituent for each carbon of a -C=C- double bond in the aliphatic chain, or up to one substituent for the carbon of a terminal methine group).

[0038] "Alkoxy" refers to the group -OR, where R is a substituted or unsubstituted alkyl or a substituted or unsubstituted cycloalkyl group. In certain instances, R is 1~6 Alkyl group or C 3~6 An exemplary alkoxy group is methoxy (-OCH 3 ) and ethoxy (-OCH 2 CH 3 In the substituted alkoxy, R is a substituted alkyl or a substituted cycloalkyl, examples of which include -OCF 2 H or -OCF 3 and the like haloalkoxy groups.

[0039] "Alkyl" is a group consisting of 1 to 25 (C 1~25 ) or more carbon atoms, more typically 1 to 10 (C 1~10 ) carbon atoms, e.g., 1 to 8 (C 1~8 ) carbon atoms, 1 to 6 (C 1~6 ) carbon atom or 1 to 4 (C 1~4 (CH ) carbon atoms. The alkyl portion can be substituted or unsubstituted. This term refers to, for example, straight chain and branched hydrocarbyl groups, such as methyl (CH 3 ), ethyl (-CH 2 CH 3 ), n-propyl (-CH 2 CH 2 CH 3 ), isopropyl (-CH(CH 3 )2 ), n-Butyl (-CH 2 CH 2 CH 2 CH 3 ), isobutyl (-CH 2 CH 2 (CH 3 ) 2 ), sec-Butyl (-CH(CH 3 )(CH 2 CH 3 ), t-Butyl (-C(CH 3 ) 3 ), n-pentyl (-CH 2 CH 2 CH 2 CH 2 CH 3 ) and neopentyl (-CH 2 C(CH 3 ) 3 As used herein, "lower alkyl" includes (C 1 ~C 8 ) alkyl.

[0040] "Aromatic" refers to a conjugated cyclic group or moiety, unless otherwise specified, having 5-15 ring atoms, either a single ring (e.g., phenyl, pyridinyl, or pyrazolyl) or multiple fused rings in which at least one ring is aromatic (e.g., naphthyl, indolyl, or pyrazolopyridinyl), i.e., at least one ring, optionally multiple fused rings, has a contiguous delocalized π-electron system. The number of out-of-plane π-electrons usually follows Hückel's rule (4n+2). The point of attachment to the parent structure is usually through an aromatic portion of the fused ring system. Examples are shown below. However, in certain instances, context, or explicit disclosure, the point of attachment may be suggested to be via a non-aromatic portion of the fused ring system. TIFF0007681570000020.tif15128 An aromatic group or moiety may contain only carbon atoms in the ring, such as, for example, an aryl group or moiety, or it may contain one or more ring carbon atoms and one or more ring heteroatoms (e.g., S, O, N, P, or Si) having an unshared electron pair, such as, for example, a heteroaryl group or moiety. Unless otherwise specified, an aromatic group may be substituted or unsubstituted.

[0041] "Aryl" refers to an aromatic carbocyclic group having 6 to 15 carbon atoms, unless otherwise specified, having a single ring (e.g., phenyl) or having multiple fused rings in which at least one ring is aromatic, multiple fused rings in which at least one ring is aromatic (e.g., 1,2,3,4-tetrahydroquinone, benzodioxole, etc.), provided that the point of attachment is through the aromatic portion of the ring system. If any portion of the aromatic ring contains a heteroatom, the group is a heteroaryl and not an aryl. Aryl groups can be, for example, monocyclic, bicyclic, tricyclic, or tetracyclic. Unless otherwise specified, aryl groups can be substituted or unsubstituted.

[0042] "Araliphatic" refers to an aryl group that is attached to its parent through an aliphatic moiety. Araliphatic groups include aralkyl or arylalkyl groups, such as benzyl and phenylethyl.

[0043] "Cyano" refers to the radical -CN.

[0044] "Alicyclic" refers to a cyclic aliphatic group having a single ring (e.g., cyclohexyl) or multiple rings, such as fused, bridged, or spirocyclic systems, at least one of which is aliphatic. Usually, the point of attachment to the parent structure is through the aliphatic portion of the multiple ring system. Alicyclic encompasses saturated and unsaturated systems, including cycloalkyl, cycloalkenyl, and cycloalkynyl. An alicyclic group can contain 3 to 25 carbon atoms; e.g., 3 to 15, 3 to 10, or 3 to 6 carbon atoms. Unless otherwise specified, an alicyclic group can be substituted or unsubstituted. Exemplary alicyclic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, or cyclohexenyl. As used herein, lower cycloalkyl refers to any of the C 3~8 Refers to cycloalkyl.

[0045] "Halo", "halide" or "halogen" refers to fluoro, chloro, bromo or iodo.

[0046] "Heteroaliphatic" refers to an aliphatic compound or group having at least one heteroatom and at least one carbon atom, i.e., an aliphatic compound or group containing at least two carbon atoms, in which at least one or more carbon atoms are replaced with an atom having at least one unshared pair of electrons, typically nitrogen, oxygen, phosphorus, silicon or sulfur. For example, a heteroalkyl moiety is a heteroaliphatic moiety, and the base aliphatic moiety is an alkyl as defined herein. A heteroaliphatic compound or group can be substituted or unsubstituted, branched or unbranched, chiral or achiral, and / or acyclic or cyclic, for example a heteroalicyclic group.

[0047] "Heteroaryl" refers to an aromatic group or moiety having 5-15 ring atoms, unless otherwise specified, and having at least one carbon atom and at least one heteroatom, such as N, S, O, P, Si. A heteroaryl group or moiety can contain a single ring (e.g., pyridinyl, pyrimidinyl, or pyrazolyl) or multiple condensed rings (e.g., indolyl, benzopyrazolyl, or pyrazolopyridinyl). A heteroaryl group or moiety can be, for example, monocyclic, bicyclic, tricyclic, or tetracyclic. Unless otherwise specified, a heteroaryl group or moiety can be substituted or unsubstituted.

[0048] "Heterocyclyl", "heterocyclo" and "heterocycle" refer to both aromatic and non-aromatic ring systems, and more specifically to stable 3- to 15-membered ring moieties containing at least one carbon atom, typically multiple carbon atoms, and at least one, e.g., 1-5, heteroatoms. The heteroatom may be a nitrogen, phosphorus, oxygen, silicon or sulfur atom. A heterocyclyl moiety may be a monocyclic moiety or may contain multiple rings, such as a bicyclic or tricyclic ring system, e.g., where at least one ring contains a heteroatom. Polycyclic moieties of this type may include spiro ring systems, in addition to fused or bridged ring systems; any of the nitrogen, phosphorus, carbon, silicon or sulfur atoms of the heterocyclyl moiety may be oxidized or may be in various oxidation states. For convenience, nitrogens, particularly (but not limited to) those defined as nitrogens in aromatic rings, are meant to include their corresponding N-oxide forms, even though specific examples do not explicitly define such. Thus, for example, in the case of a compound having a pyridinyl ring, the corresponding pyridinyl-N-oxide is included as another compound of the invention, unless expressly excluded or excluded by context. In addition, the nitrogen atom in the ring may be quaternized. Heterocycles include heteroaryl moieties (wherein the heterocyclyl moiety is aromatic), which are partially or fully saturated heterocyclyl rings, and heteroalicyclic moieties, such as heterocycloalkyl, heterocycloalkenyl, or heterocycloalkynyl.Examples of heterocyclyl groups include, but are not limited to, azetidinyl, oxetanyl, acridinyl, benzodioxolyl, benzodioxanyl, benzofuranyl, carbazoyl, cinnolinyl, dioxolanyl, indolizinyl, naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, quinazolinyl, and quinoxalinyl. , quinolinyl, isoquinolinyl, tetrazoyl, tetrahydroisoquinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, dihydropyridinyl, tetrahydropyridinyl, pyridinyl , pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl, oxazolidinyl, triazolyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl, isoindolinyl, octahydroindolyl, octahydroisoindolyl, quinolyl, isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, diazabicycloheptane, diazapane, diazepine, tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothieryl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphosphoranyl and oxadiazolyl.

[0049] "Hydroxyl" refers to the -OH group.

[0050] "Nitro" is -NO 2 Refers to the base.

[0051] "Oxo" refers to the group =O (double bonded oxygen).

[0052] "Phosphate" is -OP(O)(OR')2 In the present specification, the term refers to the group, where each -OR' is independently -OH; -O-aliphatic, e.g., -O-alkyl, or -O-cycloalkyl; -O-aromatic (including both -O-aryl and -O-heteroaryl); -O-aralkyl; or -OR' is -O - M + And M + is a counter ion having a single positive charge. + is an alkali ion, e.g., K + , Na + , Li + ammonium ion, e.g. + N(R”) 4 (wherein each R″ is independently H, aliphatic, heterocyclyl, or aryl); or an alkaline earth metal ion, such as [Ca 2+ ] 0.5 , [Mg 2+ ] 0.5 Or [Ba 2+ ] 0.5 Phosphonooxyalkyl can be an -alkyl-phosphate group, e.g., -CH 2 OP(O)(OH) 2 or a salt thereof (e.g., -CH 2 OP(O)(O - Na + ) 2 ), and (((dialkoxyphosphoryl)oxy)alkyl) refers to the dialkyl ester of a phosphonooxyalkyl group (e.g., -CH 2 OP(O)(O-tert-butyl) 2 etc.).

[0053] "Patient" or "subject" refers to mammals and other animals, particularly humans. Thus, the disclosed methods are applicable to both human therapy and veterinary applications.

[0054] "Pharmaceutically acceptable excipient" refers to a substance other than the active ingredient contained in a formulation containing the active ingredient. As used herein, an excipient may be incorporated within or physically mixed with the particles of the pharmaceutical composition. An excipient may be used, for example, to dilute the active agent and / or to modify the properties of the pharmaceutical composition. The excipient may include, but is not limited to, an anti-adherent, a binder, a coating, an enteric coating, a disintegrant, a flavoring agent, a sweetener, a coloring agent, a lubricant, a glidant, a sorbent, a preservative, an adjuvant, a carrier, or a solvent. The excipient may be starch and modified starch, cellulose and cellulose derivatives, sugars and their derivatives, such as disaccharides, polysaccharides, and sugar alcohols, proteins, synthetic polymers, cross-linked polymers, antioxidants, amino acids, or preservatives. Exemplary additives include, but are not limited to, magnesium stearate, stearic acid, vegetable stearic acid, sucrose, lactose, starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, xylitol, sorbitol, maltitol, gelatin, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), tocopheryl polyethylene glycol 1000 succinate (also known as Vitamin E TPGS or TPGS), carboxymethylcellulose, dipalmitoylphosphatidylcholine (DPPC), vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methylparaben, propylparaben, sugar, silica, talc, magnesium carbonate, sodium starch glycolate, tartrazine, aspartame, benzalkonium chloride, sesame oil, propyl gallate, sodium metabisulfite, or wool fat.

[0055] An "adjuvant" is an excipient that modifies the effect of another agent, typically an active ingredient. Adjuvants are often agents that have a pharmacological and / or immunological effect. Adjuvants can modify the effect of an active ingredient by increasing the immune response. Adjuvants can also act as stabilizers for the formulation. Exemplary adjuvants include, but are not limited to, aluminum hydroxide, alum, aluminum phosphate, killed bacteria, squalene, surfactants, cytokines, paraffin oil, and combinations of adjuvants, such as complete or incomplete Freund's adjuvant.

[0056] The term "pharmaceutically acceptable carrier" refers to an additive that is a carrier or a solvent, such as a suspending aid, a dissolving aid, or an aerosolizing aid. Pharmaceutically acceptable carriers are conventional. Remington: The Science and Practice of Pharmacy, The University of the Sciences in Philadelphia, Editor, Lippincott, Williams, &Wilkins, Philadelphia, PA, 21 st Edition (2005) describes compositions and formulations suitable for the pharmaceutical delivery of one or more therapeutic compositions and additional pharmaceutical agents.

[0057] Generally, the nature of the carrier will depend on the particular mode of administration to be used. For example, parenteral formulations usually contain medicamentously and physiologically acceptable fluids, such as injectable fluids, such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol, or solvents. In some cases, the pharmaceutically acceptable carriers can be sterilized so that they are suitable for administration to a subject (e.g., by parenteral, intramuscular, or subcutaneous injection). In addition to biologically neutral carriers, the pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents, and the like, such as sodium acetate or sorbitan monolaurate.

[0058] "Pharmaceutically acceptable salts" refers to pharma- ceutically acceptable salts of compounds derived from a variety of organic and inorganic counterions, as known to those skilled in the art, including, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc., and, where the molecule contains a basic functional group, includes salts of organic or inorganic acids, such as hydrochloride, bromide, tartrate, mesylate, acetate, maleate, oxalate, etc. "Pharmaceutically acceptable acid addition salts" are a subset of "pharmaceutically acceptable salts" that are formed with the corresponding acid but retain the biological effectiveness of the free base. In particular, the disclosed compounds include compounds that are capable of reacting with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, as well as formic acid, acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, benzenesulfonic acid, isethionic acid, salicylic acid, xinafoic acid, lactic acid, palmitic acid, alkylsulfonic acids (e.g., methanesulfonic acid, ethanesulfonic acid, 1,2- ... It forms salts with a variety of pharma- ceutically acceptable acids, including such organic acids as arylsulfonic acids (e.g., benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, and the like), 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like. Pharmaceutically acceptable salts also include salts formed when an acidic proton present in the parent compound is replaced by a metal ion (e.g., an alkali metal ion, an alkaline earth metal ion, or an aluminum ion) or coordinates with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, morpholine, piperidine, dimethylamine, diethylamine, triethylamine, ammonia, etc.).

[0059] "Pharmaceutically acceptable base addition salts" are a subset of "pharmaceutically acceptable salts" and are derived from inorganic bases, such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Exemplary salts are ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, tris(hydroxymethyl)aminomethane (tris), ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Exemplary organic bases are isopropylamine, diethylamine, tris(hydroxymethyl)aminomethane (Tris), ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine (see, e.g., SM Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19, which is incorporated herein by reference).

[0060] An "effective amount", such as a therapeutically effective amount, refers to an amount of a compound sufficient to achieve a desired result, for example, to treat a specified disorder or disease, or to reduce or eradicate one or more of its symptoms and / or prevent the occurrence of a disease or disorder. The amount of a compound that constitutes an "effective amount" may vary depending on the compound, the state of the disease and its severity, the age of the patient to be treated, and the like. An effective amount can be determined by one of ordinary skill in the art. An appropriate "effective" amount in any individual case can be determined using any suitable technique, such as a dose escalation study.

[0061] "Prodrug" refers to a compound that is converted in vivo to a biologically active compound, particularly the parent compound, which may occur, for example, by intestinal hydrolysis or enzymatic conversion. Common examples of prodrug moieties include, but are not limited to, ester and amide forms of compounds in which the activated form bears a carboxylic acid moiety. Examples of pharma- ceutically acceptable esters suitable for use with the disclosed compounds include, but are not limited to, esters of phosphate groups and carboxylic acids, such as aliphatic esters, particularly alkyl esters (e.g., C 1~6 Other prodrug moieties include phosphate esters, e.g., -CH 2 -OP(O)(OR') 2 or a salt thereof (wherein R' is H or C 1~6Further acceptable esters include cycloalkyl esters and arylalkyl esters, such as, but not limited to, benzyl. Examples of pharma- ceutically acceptable amides of the disclosed compounds include, but are not limited to, primary amides and secondary and tertiary alkyl amides (e.g., having from about 1 to about 6 carbon atoms). Amides and esters of the disclosed compounds can be prepared according to conventional methods. A thorough discussion of prodrugs is given in T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems”, Vol 14 of the ACS Symposium Series, and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference for all purposes.

[0062] "Protecting group" refers to a group of atoms that, when attached to a reactive functional group in a molecule, masks, reduces, or prevents the reactivity of the functional group. Typically, a protecting group can be selectively removed as needed during the course of a synthesis. Examples of protecting groups are described in Greene and Wuts, Protective Groups in Organic Chemistry, 3 rdEd., 1999, John Wiley & Sons, NY, and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC"), and the like. Representative hydroxyl protecting groups include, but are not limited to, those in which the hydroxyl group is acylated or alkylated, such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPPS groups), and allyl ethers.

[0063] "Solvate" refers to a complex formed when a solvent molecule combines with a molecule or ion of a solute. The solvent may be an organic compound, an inorganic compound, or a mixture of both. Some examples of solvents include, but are not limited to, methanol, ethanol, isopropanol, ethyl acetate, N,N-dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and water. The compounds described herein may exist in unsolvated as well as solvated forms when combined with pharma- ceutically acceptable or unacceptable solvents, such as water, ethanol, etc. Solvated and unsolvated forms of the compounds of the present disclosure are encompassed within the scope of the embodiments disclosed herein.

[0064] As used herein, "treat" or "treatment" refers to the treatment of CRS in a patient or subject, particularly a human suffering from CRS, including, by way of example and not limitation: (i) inhibiting CRS, e.g., stopping or slowing its progression; (ii) alleviating CRS, e.g., causing regression of CRS or its symptoms; or (iii) stabilizing CRS by preventing it from increasing in grade and / or severity; Examples include:

[0065] "Prophylaxis" as used herein relates to preventing CRS from occurring in a patient or subject, particularly where such patient or subject is at risk of developing CRS but has not yet been diagnosed as having it.

[0066] As used herein, the terms "disease" and "condition" may be used interchangeably, but may differ in that a particular disorder or condition may not have a known causative agent (and thus has yet to be determined its etiology) and is therefore merely an undesirable condition or syndrome not yet recognized as a disease, although a more or less specific set of symptoms has been identified by clinicians.

[0067] The definitions given above and the general formulae set forth below are not intended to encompass impermissible substitution patterns (e.g., methyl substituted with five fluoro groups) that would be readily recognized by one of ordinary skill in the art.

[0068] Any of the groups set forth herein may be optionally substituted with at least one, and optionally more than one, substituent as described herein, i.e., a substituted group has at least one, and optionally more than one, replaceable hydrogen atom(s) with one or more substituents as defined herein, unless the context indicates otherwise or the specific structural formula excludes substitution.

[0069] Those skilled in the art will understand that compounds may exhibit the phenomena of tautomerism, conformational isomerism, geometric isomerism, and / or optical isomerism. For example, certain disclosed compounds may contain one or more asymmetric centers and / or double bonds, and may therefore exist as stereoisomers, such as double bond isomers (i.e., geometric isomers), enantiomers, diastereomers, and mixtures thereof, such as racemic mixtures. Thus, the compounds and compositions may be provided as individual pure enantiomers or diastereomers, or as stereoisomeric mixtures, including racemic mixtures. In certain embodiments, the compounds disclosed herein are synthesized or purified to be in substantially enantiopure form, e.g., in substantially enantiopure form, e.g., with 85% enantiomeric excess (ee), 90% enantiomeric excess, 95% enantiomeric excess, 97% enantiomeric excess, 98% enantiomeric excess, 99% enantiomeric excess, or greater than 99% enantiomeric excess. One of skill in the art will understand that compounds containing one or more asymmetric centers, one or both enantiomers or diastereomers are contemplated, unless a specific enantiomer or diastereomer is shown or described.

[0070] As another example, certain disclosed compounds can exist in several tautomeric forms, such as enol forms, keto forms, and mixtures thereof. Although the various compound names, formulas, and compound drawings in this specification and claims may represent only one of the possible tautomeric, conformational, optical, or geometric isomers, those skilled in the art will understand that the disclosed compounds also encompass any tautomeric, conformational, optical, and / or geometric isomers of the compounds described herein, as well as mixtures of these various different isomers. For example, when rotation around an amide bond or between two directly bonded rings of a pyrazolyl or pyridinyl ring is restricted, atropisomers are also possible and are specifically included in the compounds of the present invention as well.

[0071] In any embodiment, any hydrogen present in the compound or in a particular group or portion of the compound may be replaced with deuterium or tritium. Thus, when alkyl is referred to, deuterated alkyl is also included, where one to the maximum number of hydrogens present may be replaced with deuterium. For example, ethyl is C 2 H 5 or C with 1 to 5 hydrogens replaced by deuterium 2 H 5 , for example C 2 D x H 5-x It could be.

[0072] II. Compounds Disclosed herein are compounds, prodrugs, corresponding salts and / or solvate forms, and methods of using these compounds, prodrugs, and salt / solvate forms for treating and / or preventing CRS. The compounds can be compounds that modulate spleen tyrosine kinase (Syk) and / or can be kinase inhibitors, such as Syk inhibitors. In some embodiments, the compounds are pyrimidinediamines represented by formula I: TIFF0007681570000021.tif28128 or a salt, solvate, N-oxide, or prodrug thereof. In some embodiments, the compound of formula I is a Syk inhibitor. With respect to formula I, Y is CH 2 , N.R. 24 , O, S, S(O) and S(O) 2 In some embodiments, Y is O.

[0073] Z 1 and Z 2 are each independently selected from CH and N. 1 is CH. In some embodiments, Z 2 is N. In certain embodiments, Z 1 is CH and Z 2 is N.

[0074] R 2 are the same or different R 8lower alkyl, optionally substituted with one or more groups, the same or different R 8 lower cycloalkyl, optionally substituted with one or more groups, the same or different R 8 cyclohexyl, which may be substituted with one or more of the following groups: 8 3-8 membered heterocycloalkyl optionally substituted with one or more groups, the same or different R 8 may be substituted with one or more of the following groups (C 6 ~C 14 ) aryl, the same or different R 8 Phenyl, optionally substituted with one or more groups, and the same or different R 8 In some embodiments, R is selected from the group consisting of 5- to 15-membered heteroaryl, optionally substituted with one or more of the following groups: 2 are the same or different R 8 and n is 1 or 2. The phenyl group is optionally substituted with one or more groups.

[0075] R 5 is selected from halo, cyano, nitro, or trihalomethyl, such as trifluoromethyl. 5 is halo and can be F.

[0076] Each R 8 are independently a , R b , the same or different R a or R b R substituted with one or more, e.g., 1 to 4, a , the same or different R a or R b -OR replaced by one or more of a , -B(OR a ) 2 , -B(NR c R c ) 2 , -(CH 2 ) m -R b , -(CHR a ) m -R b , -O-(CH 2 ) m-R b 、-S-(CH 2 ) m -R b 、-O-CHR a R b 、-O-CR a (R b ) 2 、-O-(CHR a ) m -R b 、-O-(CH 2 ) m -CH[(CH 2 ) m R b ]R b 、-S-(CHR a ) m -R b 、-C(O)NH-(CH 2 ) m -R b 、-C(O)NH-(CHR a ) m -R b 、-O-(CH 2 ) m -C(O)NH-(CH 2 ) m -R b 、-S-(CH 2 ) m -C(O)NH-(CH 2 ) m -R b 、-O-(CHR a ) m -C(O)NH-(CHR a ) m -R b 、-S-(CHR a ) m -C(O)NH-(CHR a ) m -R b 、-NH-(CH 2 ) m -R b 、-NH-(CHR a ) m -R b 、-NH[(CH 2 ) m R b ]、-N[(CH 2 ) m R b] 2 , -NH-C(O)-NH-(CH 2 ) m -R b , -NH-C(O)-(CH 2 ) m -CHR b R b , and -NH-(CH 2 ) m -C(O)-NH-(CH 2 ) m -R b In some embodiments, at least one, e.g., one, two, three, four or more R 8 is alkoxy, -O-(C 1~6 ) alkyl, for example methoxy.

[0077] R 17 is selected from hydrogen, halogen, fluoro, lower alkyl and methyl, or alternatively, R 17 is R 18 may be taken together with R to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms. 17 is C 1~6 Alkyl, for example methyl.

[0078] R 18 is selected from hydrogen, halogen, fluoro, lower alkyl and methyl, or alternatively, R 18 is R 17 may be taken together with R to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms. 18 is C 1~6 Alkyl, for example methyl.

[0079] R 19 is selected from hydrogen, lower alkyl, and methyl, or alternatively, R 19 is R 20may be taken together with the carbon atom to which they are attached to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms.

[0080] R 20 is selected from hydrogen, lower alkyl and methyl, or alternatively, R 20 is R 19 may be taken together with R to form an oxo (=O) group, or may be taken together with the carbon atom to which they are attached to form a spiro ring containing 3 to 7 carbon atoms. 19 and R 20 together form an oxo group.

[0081] Each R a are each independently hydrogen, lower alkyl, lower cycloalkyl, cyclohexyl, (C 4 ~C 11 ) cycloalkylalkyl, (C 6 ~C 10 )Aryl, phenyl, (C 7 ~C 16 ) arylalkyl, benzyl, 2- to 6-membered heteroalkyl, 3- to 8-membered heterocycloalkyl, morpholinyl, piperazinyl, homopiperazinyl, piperidinyl, 4- to 11-membered heterocycloalkylalkyl, 5- to 10-membered heteroaryl, and 6- to 16-membered heteroarylalkyl.

[0082] Each R b =O, -OR a , (C 1 ~C 3 )Haloalkyloxy, =S, -SR a , =NR a , =NOR a , -NR c R c , halogen, -CF 3 , -CN, -NC, -OCN, -SCN, -NO, -NO 2 , =N 2 , -N 3 , -S(O)R a , -S(O) 2 R a, -S(O) 2 OR a , -S(O)NR c R c , -S(O) 2 NR c R c , -OS(O)R a , -OS(O) 2 R a , -OS(O) 2 OR a , -OS(O) 2 NR c R c , -C(O)R a , -C(O)OR a , -C(O)NR c R c , -C(NH)NR c R c , -C(NR a )NR c R c , -C(NOH)R a , -C(NOH)NR c R c , -OC(O)R a , -OC(O)OR a , -OC(O)NR c R c , -OC(NH)NR c R c , -OC(NR a )NR c R c , -[NHC(O)] n R a , -[NR a C(O)] n R a , -[NHC(O)] n OR a , -[NR a C(O)] n OR a , -[NHC(O)] n NR c R c , -[NR a C(O)] n NR c R c , -[NHC(NH)] n NR c R c and -[NRa C(NR a )] n NR c R c are suitable groups independently selected from:

[0083] Each R c are each independently an amino protecting group and R a or alternatively, two R c may, together with its nitrogen atom, contain one or more of the same or different additional heteroatoms and may be the same or different R a It forms a 5-8 membered heterocycloalkyl or heteroaryl which may be optionally substituted with one or more, for example 1 to 4, groups.

[0084] The amino protecting group can be any protecting group suitable to act as a protecting group for an amine moiety. In some embodiments, the amino protecting group is formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl ("CBZ"), tert-butoxycarbonyl ("Boc"), trimethylsilyl ("TMS"), 2-trimethylsilyl-ethanesulfonyl ("TES"), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl ("FMOC"), nitro-veratryloxycarbonyl ("NVOC"), and the like.

[0085] R 21 , R 22 and R 23 are each independently selected from hydrogen and phosphonooxyalkyl.

[0086] R 24 is selected from hydrogen, lower alkyl, and phosphonooxyalkyl.

[0087] Each m is independently an integer from 1 to 3.

[0088] Each n is independently an integer from 0 to 3.

[0089] In some embodiments, R 21 , R 22 , R 23 and R 24 is phosphonooxyalkyl, but in other embodiments, R 21 , R 22 , R 23 and R 24 is not phosphonooxyalkyl. In certain embodiments, R 21 , R 22 , R 23 and R 24 Each of R is hydrogen. 21 is phosphonooxyalkyl, R 22 , R 23 and R 24 Each of is hydrogen.

[0090] In some embodiments, the compound of formula I can be represented by formula II, III or IV TIFF0007681570000022.tif67129, or a salt, solvate, N-oxide, or prodrug thereof, wherein Y, Z 2 , R 2 , R 5 , R 8 , R 17 , R 18 , R 19 , R 20 , and R 21 is as defined for formula I, if present.

[0091] In some embodiments, the phosphonooxyalkyl moieties I-IV are -CH 2 OP(=O)(R 31 ) 2 where each R 31 are independently -OH, -O-aliphatic, e.g., -O-alkyl, or -O - M + And M + is a counterion that carries one positive charge.

[0092] In some embodiments, the compound has formula V TIFF0007681570000023.tif30128, or a salt, solvate, N-oxide, or prodrug thereof, wherein R 30 is H or phosphonooxyalkyl. In some embodiments, R 30 is H, but in other embodiments, R 30 -CH 2 OP(=O)(R 31 ) 2 where each R 31 are independently -OH, -O-aliphatic, e.g., -O-alkyl, or -O - M + And M + is a counterion that carries one positive charge.

[0093] Exemplary compounds represented by one or more of formulas I-XI are: Contains TIFF0007681570000024.tif61147.

[0094] The FDA has approved TAVALISSE® for the treatment of thrombocytopenia in adult patients with immune thrombocytopenia (ITP) who have had an inadequate response to previous therapy. The active agents in TAVALISSE® are fostamatinib disodium hexahydrate, [6-[[5-fluoro-2-(3,4,5-trimethoxyanilino)pyrimidin-4-yl]amino]-2,2-dimethyl-3-oxopyrido[3,2-b][1,4]oxazin-4-yl]methyl phosphate disodium hexahydrate, TIFF0007681570000025.tif35128Fostamatinib (R788), disodium [6-({5-fluoro-2-[(3,4,5-trimethoxyphenyl)amino]-4-pyrimidinyl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b][1,4]oxazin-4-yl]methyl dihydrogen phosphate hexahydrate, The file is TIFF0007681570000026.tif35128.

[0095] Fostamatinib and its disodium hexahydrate salt are the active metabolites 6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazin-3(4H)-one (R406), It is a prodrug of TIFF0007681570000027.tif21128.

[0096] R406 is an inhibitor of the enzyme spleen tyrosine kinase (Syk), which plays a key role in signal transduction of activated Fc receptors and B cell receptors (BCR). Syk is involved in signal transduction associated with the high affinity Fc receptors for IgE ("FcεRI") and / or IgG ("FcγRI") (for review, see Valent et al., 2002, Intl. J. Hematol. 75(4):257-362). Biochemical data confirm that 2,4-pyrimidinediamine compounds such as R406 exert their degranulation inhibitory effects, at least in part, by blocking or inhibiting the signal transduction cascade initiated by crosslinking of FcεRI and / or FcγRI (see, e.g., U.S. Patent Application Serial No. 10 / 631,029, filed July 29, 2003 (U.S. Patent Application Publication No. 2007 / 0060603, now U.S. Patent No. 7,517,886, filed July 29, 2003). No. PCT / US03 / 24087 (WO 2004 / 014382), U.S. Patent Application Serial No. 10 / 903,263, filed July 30, 2004 (U.S. Patent Application Publication No. 2005 / 0234049), and International Application No. PCT / US2004 / 24716 (WO 2005 / 016893), the disclosures of which are incorporated herein by reference.

[0097] Braselmann et al. (J. Pharmacol. Exp. Ther. 319(3):998-1008 (Dec. 2006; epub Aug. 31, 2006) demonstrate that R406 is a potent inhibitor of IgE- and IgG-mediated activation of Fc receptor signaling (EC 50 =56-64 nM). R406 inhibited phosphorylation of the Syk substrate linker for activation of T cells in mast cells and B cell linker protein / SLP65 in B cells. R406 bound to the ATP-binding pocket of Syk and inhibited its kinase activity as an ATP-competitive inhibitor (K(i)=30 nM). Furthermore, R406 blocked Syk-dependent FcR-mediated activation of monocytes / macrophages and neutrophils and BCR-mediated activation of B lymphocytes.

[0098] As used herein, "form of fostamatinib" refers to fostamatinib, a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable salt hydrate thereof, or a Syk-inhibiting metabolite thereof. In one embodiment, the form of fostamatinib is fostamatinib disodium hexahydrate (e.g., TAVALISSE®). In another embodiment, the form of fostamatinib is metabolite R406.

[0099] Disclosed herein are methods of blocking or ameliorating treatment-induced CRS by administering a form of fostamatinib (e.g., fostamatinib or a therapeutically acceptable salt thereof, e.g., fostamatinib disodium hexahydrate, and R406). The inventors have unexpectedly found that a form of fostamatinib can block or ameliorate CRS toxicity or therapeutic rescue treatment in combination with steroids to reduce toxicity. A form of fostamatinib advantageously reduces toxicity resulting from CRS while maintaining the efficacy of the immunotherapy / administered cells.

[0100] Further information regarding compounds of formulas IX can be found in US Pat. Nos. 7,449,458 and 8,163,902, which are incorporated by reference in their entireties.

[0101] IV. Compositions Containing Compounds Disclosed Herein The disclosed compounds may be used alone or in combination, and / or in conjunction with or adjunctively with at least one second therapeutic agent, and additional compounds, and at least one second therapeutic agent, if present, may be used in combination with any suitable additive useful in forming a composition for administration to a subject. Additives may be included in pharmaceutical compositions for a variety of purposes, such as diluting the composition for delivery to a subject, facilitating processing of the formulation, imparting advantageous material properties to the formulation, facilitating dispersion from a delivery device, stabilizing the formulation (e.g., antioxidants or buffers), imparting a pleasant or palatable taste or consistency to the formulation, etc.Typical excipients include, by way of example and not limitation: carriers and / or adjuvants, such as mono-, di- and polysaccharides, sugar alcohols and other polyols, such as lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitol, diphosphatidylcholine, and lecithin; bulking agents; buffers, such as phosphate and citrate buffers; anti-adherents, such as magnesium stearate; binders, such as sugars (including disaccharides, such as sucrose and lactose), polysaccharides (such as starch, cellulose, microcrystalline cellulose, cellulose ethers (such as hydroxypropyl cellulose), gelatin, synthetic polymers (such as polyvinylpyrrolidone, polyalkylene glycols); coatings (such as cellulose ethers such as hydroxypropyl methylcellulose, shellac, corn protein zein, and gelatin). ; release aids (e.g., enteric coatings); disintegrants (e.g., crospovidone, cross-linked sodium carboxymethylcellulose, and sodium starch glycolate); fillers (e.g., dicalcium phosphate, vegetable fats, lactose, sucrose, glucose, mannitol, sorbitol, calcium carbonate, and magnesium stearate); flavorings and sweeteners (e.g., mint, cherry, anise, peach, apricot or licorice, raspberry, and vanilla; lubricants (e.g., minerals such as talc or silica, fats such as vegetable stearin, magnesium stearate, or stearic acid); preservatives (e.g., antioxidants such as vitamin A, vitamin E, vitamin C, retinyl palmitate, and selenium, amino acids such as cysteine ​​and methionine, citric acid and sodium citrate, parabens such as methylparaben and propylparaben); colorants; compression aids; emulsifiers; encapsulating agents; gums; granulating agents; and combinations thereof.

[0102] V. Therapeutic Combinations The disclosed compounds may be used alone, in combination with other disclosed compounds, and / or adjunctively or in combination with other established therapies. In another embodiment, the compounds may be used in combination with other therapeutic agents useful for treating CRS, and / or other diseases or conditions. The compounds and / or other agents may be administered simultaneously, sequentially in any order, by the same route of administration, or by different routes.

[0103] In some embodiments, the second therapeutic agent is an analgesic, an antibiotic, an anticoagulant, an antibody, an anti-inflammatory agent, an immunosuppressant, a guanylate cyclase-C agonist, an intestinal secretagogue, an antiviral agent, an anticancer agent, an antifungal agent, or a combination thereof. In certain embodiments, the second therapeutic agent can be an anti-inflammatory agent, an immunosuppressant, and / or a steroid.

[0104] The anti-inflammatory agent may be a steroid, such as budesonide, or a non-steroidal anti-inflammatory agent. In certain embodiments, the non-steroidal anti-inflammatory agent is selected from aminosalicylates (e.g., sulfasalazine, mesalamine, olsalazine, and balsalazide), cyclooxygenase inhibitors (COX-2 inhibitors, such as rofecoxib, celecoxib), diclofenac, etodolac, famotidine, fenoprofen, flurbiprofen, ketoprofen, ketorolac, ibuprofen, indomethacin, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, salsalate, sulindac, tolmetin, or a combination thereof.

[0105] In some embodiments, the immunosuppressant is mercaptopurine; corticosteroids such as dexamethasone, hydrocortisone, prednisone, methylprednisolone, and prednisolone; alkylating agents such as cyclophosphamide; calcineurin inhibitors such as cyclosporine, sirolimus, and tacrolimus; inhibitors of inosine monophosphate dehydrogenase (IMPDH) such as mycophenolic acid, mycophenolate mofetil, and azathioprine; and agents designed to preserve the recipient's humoral immune response intact while suppressing cellular immunity, including various antibodies (e.g., antilymphocyte globulin (ALG), antithymocyte globulin (ATG), monoclonal anti-T cell antibody (OKT3)), and radiation; or combinations thereof. In one embodiment, the antibody is infliximab. Azathioprine is currently available from Salix Pharmaceuticals, Inc. under the trade name Azasan; mercaptopurine is currently available from Gate Pharmaceuticals, Inc. under the trade name Purinethol; prednisone and prednisolone are currently available from Roxane Laboratories, Inc; methylprednisolone is currently available from Pfizer; sirolimus (rapamycin) is currently available from Wyeth-Ayerst under the trade name Rapamune; tacrolimus is currently available from Fujisawa under the trade name Prograf; cyclosporine is currently available from Novartis under the trade name Sandimmune and from Abbott under the trade name Gengraf; IMPDH inhibitors such as mycophenolate mofetil and mycophenolic acid are currently available from Roche under the trade name Cellcept and from Novartis under the trade name Myfortic; azathioprine is currently available from Glaxo Smith It is available from Kline under the trade name Imuran; antibodies are currently available from Ortho Biotech under the trade name Orthoclone, from Novartis under the trade name Simulect (basiliximab), and from Roche under the trade name Zenapax (daclizumab).

[0106] In certain embodiments, the second therapeutic agent is or includes a steroid, e.g., a corticosteroid, including, but not limited to, a glucocorticoid and / or a mineralocorticoid. Suitable steroids for use in combination with the disclosed compounds include synthetic and non-synthetic glucocorticoids. Exemplary steroids, e.g., glucocorticoids, suitable for use in the disclosed methods include, but are not limited to, alclomethasone, algestone, beclomethasone (e.g., beclomethasone dipropionate), betamethasone (e.g., betamethasone 17-valerate, betamethasone sodium acetate, betamethasone sodium phosphate, betamethasone valerate), budesonide, clobetasol (e.g., clobetasol propionate), clobetasone , clocortolone (e.g., clocortolone pivalate), cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone (e.g., dexamethasone 21-phosphate, dexamethasone acetate, dexamethasone sodium phosphate), diflorasone (e.g., diflorasone diacetate), diflucortolone, difluprednate, enoxolone, fluazacort acort), flucloronide, fludrocortisone (e.g., fludrocortisone acetate), flumethasone (e.g., flumethasone pivalate), flunisolide, fluocinolone (e.g., fluocinolone acetonide), fluocinonide, fluocortin, fluocortolone, fluorometholone (e.g., fluorometholone acetate), fluperolone (e.g., fluperol acetate), fluprednidene, fluprednisolone, flurandrenolide, flu fluticasone (e.g., fluticasone propionate), formocortal, halcinonide, halobetasol, halometasone, halopredone, hydrocortamate, hydrocortisone (e.g., hydrocortisone 21-butyrate, hydrocortisone aceponate, hydrocortisone acetate, hydrocortisone buteprate, hydrocortisone butyrate, hydrocortisone cypionate, hydrocortisone hemisuccinate, hydrocortisone probutate, hydrocortisone sodium phosphate,hydrocortisone sodium succinate, hydrocortisone valerate), loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone (methylprednisolone aceponate, methylprednisolone acetate, methylprednisolone hemisuccinate, methylprednisolone sodium succinate), mometasone (e.g., mometasone furoate), paramethasone (e.g., paramethasone acetate), prednicarbate, prednisolone (e.g., prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisolone 21-hemisuccinate, acetate Prednisolone; prednisolone farnesylate, prednisolone hemisuccinate, prednisolone-21 (β-D-glucuronide), prednisolone metasulfobenzoate, prednisolone stearate, prednisolone tebutate, prednisolone tetrahydrophthalate), prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone (e.g., triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, triamcinolone acetonide 21-palmitate, triamcinolone diacetate), or any combination thereof. Further information regarding steroids and their salts can be found, for example, in Remington's Pharmaceutical Sciences, A. Osol, ed., Mack Pub. Co., Easton, Pa. (16th ed. 1980).

[0107] In some cases, the steroid is a glucocorticoid and may be selected from cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, or combinations thereof. In certain instances, the steroid is or includes prednisone. In other specific instances, the steroid is or includes dexamethasone.

[0108] VI. Formulation and Administration Pharmaceutical compositions containing one or more of the disclosed compounds (including salts, solvates, N-oxides and / or prodrugs thereof) can be prepared by conventional mixing, dissolving, granulating, sugar-coating, mashing, emulsifying, encapsulating, entrapping or lyophilizing processes. The compositions can be formulated in a conventional manner with one or more physiologically acceptable excipients, diluents, carriers, adjuvants or auxiliary agents to obtain medicamentally usable preparations. A wide variety of suitable pharmaceutical compositions are known in the art. See, for example, Remington: The Science and Practice of Pharmacy, volume I and volume II. (22 nd Ed., University of the Sciences, Philadelphia).

[0109] The disclosed compounds, or prodrugs thereof, can be formulated in pharmaceutical compositions themselves, or in the form of solvates, N-oxides, or pharma- ceutically acceptable salts.Typically, such salts are more soluble in aqueous solutions than the corresponding free acids and bases, although salts that are less soluble than the corresponding free acids and bases can also be formed.

[0110] Pharmaceutical compositions containing one or more of the disclosed compounds can be in a form suitable for virtually any mode of administration, including, for example, topical, ocular, oral, buccal, systemic, nasal, injection (such as intravenous or intraperitoneal), transdermal, rectal, vaginal, sublingual, urethral (e.g., urethral suppository), or a form suitable for inhaled or insufflated administration. In certain embodiments, the mode of administration is oral or injection.

[0111] Systemic formulations include those designed for administration by injection, e.g., subcutaneous, intravenous, intramuscular, intrathecal or intraperitoneal injection, and those designed for transdermal, transmucosal, oral or pulmonary administration.

[0112] Useful injection preparations include sterile suspensions, solutions or emulsions of active compounds in aqueous or oily solvents.The compositions may also contain formulation aids such as suspending agents, stabilizing agents and / or dispersing agents.Injection preparations may be provided in unit dosage form, for example, in ampoules or in multi-dose containers, and may contain added preservatives.

[0113] Alternatively, the injectable formulations may be provided in powder form for reconstitution with a suitable solvent (including but not limited to sterile pyrogen-free water, buffer, dextrose solution, etc.) before use. To this end, the disclosed compounds can be dried by any known technique, such as lyophilization, and reconstituted prior to use.

[0114] For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are known in the art.

[0115] For oral administration, the pharmaceutical compositions may take the form of, for example, lozenges, tablets, or capsules prepared by conventional means with pharma- ceutically acceptable additives, such as binders (e.g., pregelatinized maize starch, polyvinylpyrrolidone, or hydroxypropylmethylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium starch glycolate); and / or wetting agents (e.g., sodium lauryl sulfate). Tablets may be coated, for example, with sugar, film, or enteric coatings, by methods well known in the art.

[0116] Furthermore, pharmaceutical compositions containing the disclosed compounds or their solvates, N-oxides, pharma- ceutically acceptable salts, or prodrugs as active ingredients in a form suitable for oral use may also include, for example, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide a medicamentously elegant and palatable preparation. Tablets contain the active ingredient (including prodrugs) in admixture with non-toxic pharma- ceutical acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents (e.g., corn starch, or alginic acid); binding agents (e.g., starch, gelatin, or acacia); and lubricants (e.g., magnesium stearate, stearic acid, or talc). The tablets can be uncoated or they can be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated by the techniques described in U.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotic therapeutic tablets for controlled release. The pharmaceutical compositions of the invention can also be in the form of oil-in-water emulsions. Tablets may also be film coated, which may include one or more of polyvinyl alcohol, titanium dioxide, polyethylene glycol 3350, talc, yellow iron oxide, and red iron oxide.

[0117] Liquid preparations for oral administration can take the form of, for example, elixirs, solutions, syrups or suspensions, or can be provided as a dry product for constitution with water or other suitable solvent before use. Such liquid preparations can be prepared by conventional means with pharma- ceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or gum arabic); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, cremophore™ or fractionated vegetable oils); and preservatives (e.g., p-hydroxybenzoic acid or methyl or propyl sorbate). The preparations can further contain buffer salts, preservatives, flavoring agents, coloring agents, and sweetening agents, if appropriate.

[0118] Preparations for oral administration can be suitably formulated to give controlled release of the disclosed compounds, as is known in the art.

[0119] For buccal administration, the compositions can take the form of tablets or lozenges formulated in conventional manner.

[0120] For topical administration, the disclosed compounds (including solvates, N-oxides, or pharma- ceutically acceptable salts and / or prodrugs thereof) may be formulated as solutions, gels, ointments, creams, suspensions, etc., as are well known in the art.

[0121] For rectal and vaginal administration routes, the active compounds can be formulated as solutions (for retention enemas), suppositories, or ointments containing conventional suppository bases such as cocoa butter or other glycerides.

[0122] For nasal administration or administration by inhalation or insufflation, the disclosed compounds, solvates, N-oxides, pharma- ceutically acceptable salts or prodrugs can be conveniently delivered in the form of an aerosol spray from a pressurized charge or nebulizer using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges (e.g., capsules and cartridges comprised of gelatin) for use in an inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0123] The pharmaceutical composition may be in the form of a sterile injectable aqueous or oily suspension. This suspension may be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents as mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution.

[0124] In accordance with the present invention, certain forms of the disclosed compounds, solvates, N-oxides, pharma- ceutical acceptable salts or prodrugs thereof, can also be used in the preparation of compounds for use in the manufacture of pharmaceutical compositions, e.g., U.S. Pat. No. 6,241,969; U.S. Pat. No. 6,060,069; U.S. Pat. No. 6,238,647; U.S. Pat. No. 6,335,316; U.S. Pat. No. 5,364,838; U.S. Pat. No. 5,672,581; WO 96 / 32149; WO 95 / 24183; U.S. Pat. No. 5,654,007; U.S. Pat. No. 5,404,871; U.S. Pat. No. 5,672,581; U.S. Pat. No. 5,743,250; U.S. Pat. No. 5,419,315; U.S. Pat. No. 5,558,085; WO 98 / 33480; U.S. Pat. No. 5,364,833 No. 5,320,094; No. 5,780,014; No. 5,658,878; No. 5,518,998; No. 5,506,203; No. 5,661,130; No. 5,655,523; No. 5,645,051; No. 5,622,166; No. 5,577,49 The compositions may be delivered by any of a variety of inhalation devices and methods known in the art, including those disclosed in U.S. Patent Publication No. 20010041190; U.S. Patent Publication No. 20020006901; and U.S. Patent Publication No. 20020034477.

[0125] Among the devices that can be used to administer a form of the active compound are those well known in the art, such as metered dose inhalers, liquid nebulizers, dry powder inhalers, nebulizers, thermal vaporizers, etc. Other suitable techniques for administration of certain 2,4-pyrimidinediamine compounds include electrohydrodynamic aerosolizers.

[0126] Moreover, the inhalation device is preferably practical in the sense that it is easy to use, small enough to be conveniently carried, capable of providing multiple doses, and durable.Some specific examples of commercially available inhalation devices are Turbohaler (Astra, Wilmington, DE), Rotahaler (Glaxo, Research Triangle Park, NC), Diskus (Glaxo, Research Triangle Park, NC), Ultravent nebulizer (Mallinckrodt), Acorn II nebulizer (Marquest Medical Products, Totowa, NJ), Ventolin metered dose inhaler (Glaxo, Research Triangle Park, NC), etc. In one embodiment, the disclosed compounds, their solvates, N-oxides, pharma- ceutically acceptable salts, or prodrugs can be delivered by dry powder inhalers or nebulizers.

[0127] As those skilled in the art will recognize, the formulation of the disclosed compound, its solvates, N-oxides, pharmaceutically acceptable salts or prodrugs, the amount of the formulation delivered, and the duration of administration of a single dose depend on the type of inhalation device used as well as other factors. In some aerosol delivery systems, such as nebulizers, the frequency of administration and the length of time the system is activated will depend primarily on the disclosed compound in the aerosol. For example, a shorter duration of administration can be used with a higher concentration of the disclosed compound in the nebulizer solution. Devices such as metered dose inhalers can produce higher aerosol concentrations and can be operated for a shorter period of time to deliver a desired amount of active compound in some embodiments. Devices such as dry powder inhalers deliver active drug until a given charge of drug is released from the device. In this type of inhaler, the amount of the disclosed compound, its solvates, N-oxides, pharmaceutically acceptable salts or prodrugs in a given amount of powder determines the dose delivered in a single administration. The formulation of the disclosed compound is selected to obtain the desired particle size in the selected inhalation device.

[0128] Formulations of the disclosed compounds, such as fostamatinib, for administration from a dry powder inhaler typically include a finely divided dry powder containing the disclosed compounds, although the powder may also include bulking agents, buffers, carriers, excipients, other additives, etc. Additives may be included in the dry powder formulation, for example, to dilute the powder when needed for delivery from a particular powder inhaler, to facilitate processing of the formulation, to impart advantageous powder properties to the formulation, to facilitate dispersion of the powder from the inhalation device, to stabilize the formulation (e.g., antioxidants or buffers), to impart flavor to the formulation, etc. Typical additives include mono-, di-, and polysaccharides; sugar alcohols and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch, or combinations thereof; surfactants, such as sorbitol, diphosphatidylcholine, or lecithin.

[0129] The method of the present invention can be carried out for pharmaceutical compositions comprising the disclosed compounds suitable for administration by inhalation, such as fostamatinib. For example, dry powder formulations can be produced in a number of ways using conventional techniques, such as those described in any of the publications mentioned above, which are expressly incorporated herein by reference, and, for example, in U.S. Patent No. 5,700,904 to Baker et al., the entire disclosure of which is expressly incorporated herein by reference. Particles of a size range suitable for maximum deposition in the lower respiratory tract can be produced by micronization, milling, etc., and liquid formulations can be produced by dissolving the compound in a suitable solvent, such as water, at an appropriate pH, including buffers or other excipients.

[0130] A specific example of an aqueous suspension formulation suitable for nasal administration using a commercially available nasal spray contains the following ingredients: active compound or prodrug (0.5-20 mg / mL); benzalkonium chloride (0.1-0.2 mg / mL); polysorbate 80 (TWEEN® 80; 0.5-5 mg / mL); sodium carboxymethylcellulose or microcrystalline cellulose (1-15 mg / mL); phenylethanol (1-4 mg / mL); and dextrose (20-50 mg / mL). The pH of the final suspension can be adjusted to a range of about pH 5 to pH 7, with a typical pH being about pH 5.5.

[0131] Another specific example of an aqueous suspension suitable for administration of a compound by inhalation contains compound or prodrug 20 mg / mL, polysorbate 80 (TWEEN® 80) 1% (v / v), citrate 50 mM, and / or sodium chloride 0.9%.

[0132] For intraocular administration, the active compound or prodrug can be formulated into a solution, emulsion, suspension, etc. suitable for intraocular administration. A variety of solvents suitable for intraocular administration of compounds are known in the art. Non-limiting examples are described in U.S. Patent No. 6,261,547; U.S. Patent No. 6,197,934; U.S. Patent No. 6,056,950; U.S. Patent No. 5,800,807; U.S. Patent No. 5,776,445; U.S. Patent No. 5,698,219; U.S. Patent No. 5,521,222; U.S. Patent No. 5,403,841; U.S. Patent No. 5,077,033; U.S. Patent No. 4,882,150; and U.S. Patent No. 4,738,851, which are incorporated herein by reference.

[0133] In order to deliver the disclosed compounds continuously, they can be formulated as depot preparations for administration by implantation or intramuscular injection.The active ingredient can be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in acceptable oils) or with ion exchange resins, or as poorly soluble derivatives, for example, poorly soluble salts.Alternatively, a transdermal delivery system can be used, which is manufactured as a circular patch or patch that slowly releases the disclosed compounds to be absorbed through the skin.For this purpose, a penetration enhancer can be used to promote the percutaneous penetration of the active compound. Suitable transdermal patches are described, for example, in U.S. Pat. Nos. 5,407,713; 5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189; 5,163,899; 5,088,977; 5,087,240; 5,008,110; and 4,921,475, which are incorporated herein by reference.

[0134] Alternatively, other pharmaceutical delivery systems can be used. Liposomes and emulsions are examples of well-known delivery vehicles that can be used to deliver active compounds or prodrugs. Certain organic solvents such as dimethylsulfoxide (DMSO) can also be used, usually at the expense of higher toxicity. In some embodiments, the disclosed compounds as active ingredients or their solvates, N-oxides, pharma-ceutically acceptable salts or prodrugs are orally administered in the form of tablets, for example, a form of fostamatinib can be administered as TAVALISSE®.

[0135] The pharmaceutical composition may be provided in a package or dispenser that may contain one or more unit dosage forms containing the active compound, if desired. The package may, for example, comprise metal or plastic foil, such as a blister pack. The package or dispenser may also be accompanied by instructions for administration.

[0136] VII. Dosage The disclosed compounds or compositions thereof will generally be used in an amount effective to achieve a desired result, for example, an amount effective to treat or prevent CRS. The compounds or compositions thereof can be administered therapeutically to achieve a therapeutic effect and / or prophylactically to achieve a prophylactic effect. By therapeutic effect is meant that the underlying CRS is eradicated or alleviated and / or one or more symptoms associated with CRS are eradicated or alleviated such that the patient reports an improved feeling or condition, regardless of whether the patient may still be suffering from CRS. In some embodiments, an indication of therapeutic improvement and / or successful treatment may include preventing the subject from exhibiting one or more symptoms in the relevant score on the CRS rating scale, for example, preventing the subject from exhibiting CRS of grade 2 or higher. Additionally or alternatively, an indication of therapeutic improvement and / or successful treatment may be a change in the rating or severity on the rating scale described herein, for example, a change from a score of 4 to a score of 3 or less, or a change from a score of 3 to a score of 2 or 1. A prophylactic effect may be achieved by substantially preventing CRS from developing, e.g., by preventing the onset of any symptoms or by preventing one or more symptoms from progressing beyond grade 1. In some embodiments, a prophylactic effect may mean preventing a subject from exhibiting one or more symptoms at a level of grade 2 or higher.

[0137] As known by those skilled in the art, the preferred dosage of the compound may also depend on a variety of factors, including the age, weight, general health, and severity of the condition of the patient or subject being treated. The dosage may also need to be adjusted to the gender of the individual and / or the lung volume of the individual when administered by inhalation. The dosage may also be adjusted for individuals suffering from more than one condition or individuals with additional conditions that adversely affect lung volume and breathing capacity, such as emphysema, bronchitis, pneumonia, and respiratory infections in general. The dosage and frequency of administration of the disclosed compounds or compositions thereof will also depend on whether the compound is prescribed for the treatment of an acute episode of CRS or for the prophylactic treatment of CRS. Those skilled in the art will be able to determine the optimal dose for a particular individual.

[0138] The disclosed compounds, or compositions thereof, may be administered before, during, and / or after a treatment that may induce CRS. In one embodiment, the disclosed compounds, or compositions thereof, may be administered within 48 hours before the start of a treatment that may induce CRS, such as within 24, 12, 6, 4, or 2 hours of the treatment. In another embodiment, the disclosed compounds, or compositions thereof, may be administered during the course of a treatment. In another embodiment, the disclosed compounds, or compositions thereof, may be administered after the completion of a treatment, immediately or shortly after the completion of the treatment (e.g., within 24, 48, 72, or 96 hours or one week of the completion of the treatment). In another embodiment, the disclosed compounds, or compositions thereof, may be administered in two or more of the periods consisting of before, during, or after the treatment.

[0139] For prophylactic administration, the disclosed compounds, or compositions thereof, can be administered to patients or subjects at risk of developing CRS. For example, the compounds, or compositions thereof, can be administered to a subject prior to the initiation of a treatment likely to cause CRS, substantially simultaneously with the initiation of such a treatment, or after the treatment has been initiated. The compounds, or compositions thereof, can also be administered prophylactically to an individual who may be repeatedly treated with a treatment that has caused CRS in other individuals, even if the subject has not previously developed CRS.

[0140] An effective dose can be estimated initially from in vitro testing. For example, an initial dose for use in a subject can be determined based on the IC200 of a particular compound determined in an in vitro test, as determined by circulating blood or serum concentrations of the active compound. 50 or EC 50 The compound can be formulated to reach or exceed the effective dose. The dose can be calculated to reach such a circulating or serum concentration, taking into account the bioavailability of the particular compound. Further guidance regarding effective dosages can be found in Fingl & Woodbury, "General Principles," In: Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, Chapter 1, pages 1-46, Pergamon Press and references cited therein.

[0141] In some embodiments, the EC of the disclosed compounds 50 is greater than 0 to 20 μM, for example, greater than 0 to 10 μM, greater than 0 to 5 μM, greater than 0 to 1 μM, greater than 0 to 0.5 μM, or greater than 0 to 0.1 μM.

[0142] Initial doses can also be estimated from in vivo data, such as animal models, including mouse and non-human primate models. CRS animal models are known to those of skill in the art, and further information can be found in Norelli, M., Camisa, B., Barbiera, G. et al. Monocyte-derived IL-1 and IL-6 are differentially required for cytokine-release syndrome and neurotoxicity due to CAR T cells. Nat Med. 2018;24:739-748, and Giavridis, T., van der Stegen, SJC, Eyquem, J., Hamieh, M., Piersigilli, A., and Sadelain, M. CAR T cell-induced cytokine release syndrome is mediated by macrophages and abated by IL-1 blockade. Nat Med. 2018;24:731-738.

[0143] Dosages of the disclosed compounds typically range from greater than about 0 mg / kg / day, such as 0.0001 mg / kg / day or 0.001 mg / kg / day or 0.01 mg / kg / day, up to at least about 1000 mg / kg / day, such as up to 100 mg / kg / day, but can be higher or lower depending on, among other factors, the activity of the compound, its bioavailability, the mode of administration, and various factors described herein. More typically, dosages (or effective amounts) can be administered at least once per day in the range of about 0.0025 mg / kg to about 1 mg / kg, such as 0.01 mg / kg to about 0.5 mg / kg or about 0.05 mg / kg to about 0.15 mg / kg. The total daily dosage is usually in the range of about 0.1 mg / kg / day to about 5 mg / kg / day or about 20 mg / kg / day, for example, 0.5 mg / kg / day to about 10 mg / kg / day or about 0.7 mg / kg / day to about 2.5 mg / kg / day. Dosages can be higher or lower depending on, among other factors, the activity of the compound, its bioavailability, the mode of administration, and various factors mentioned above.

[0144] The dosage amount and the interval of administration can be adjusted to obtain a plasma concentration sufficient to achieve and / or maintain the desired therapeutic or prophylactic effect of the compound for an individual. For example, the compound can be administered once a day, multiple times a day, once a week, multiple times a week (e.g., every other day), once a month, multiple times a month, or once a year, depending on, among other things, the mode of administration, the specific target disease being treated, and the judgment of the prescribing physician. Those skilled in the art will be able to optimize the effective local dosage without undue experimentation. In some embodiments, the amount of the disclosed compound in the composition administered or the amount of the compound administered in the method disclosed herein is a suboptimal dose. As used herein, a suboptimal dose is a dose typically used in a single administration to a patient in monotherapy or in combination with standard treatment.

[0145] Compositions containing one or more disclosed compounds typically contain from greater than 0 to 99% by weight of the compound(s) and / or other therapeutic agent(s). More typically, compositions containing one or more disclosed compounds contain from about 1 to about 20 percent by weight of the compounds and other therapeutic agents in total, and from about 80 to about 99 percent by weight of pharma- ceutically acceptable excipients.

[0146] A typical daily dose can range from 100 to 300 mg / day, e.g., 100, 150, 200, 250, or 300 mg / day. Dosing can be once or twice a day, or more, e.g., 100 or 150 mg BID. Thus, a pharmaceutical dosage form comprising a compound disclosed herein can contain 50 to 300 mg of the disclosed compound, e.g., 50, 100, 150, 200, 250, 300 mg of the disclosed compound. In one non-limiting embodiment, the dosage is 100 mg of the disclosed compound, e.g., in tablet form, of fostamatinib (e.g., TAVALISSE®). In another non-limiting embodiment, the dosage is 150 mg of the disclosed compound, e.g., in tablet form, of fostamatinib (e.g., TAVALISSE®).

[0147] In one embodiment, the methods of the invention are carried out by initiating administration of a disclosed compound, e.g., in the form of fostamatinib (e.g., TAVALISSE®), at 100 mg orally twice daily, with or without food. After four weeks, administration is increased to 150 mg twice daily as needed to reduce the risk of bleeding and achieve a platelet count of at least 50×109 / L.

[0148] Preferably, the compound or composition thereof will exhibit therapeutic or prophylactic effects without significant toxicity. The toxicity of a compound can be determined using standard pharmaceutical procedures. The ratio of the dose exhibiting a toxic effect and the dose exhibiting a therapeutic (or prophylactic) effect is the therapeutic index. Compounds exhibiting a high therapeutic index are preferred. EXAMPLES

[0149] VIII. Examples Example 1 N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3) TIFF0007681570000028.tif121128N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (1, 1.0 g, 2.12 mmol) in acetone (20 mL), Cs 2 CO 3 (1.0 g, 3.07 mmol) and di-tert-butyl chloromethyl phosphate (2, 0.67 g, 2.59 mmol) were stirred at room temperature under a nitrogen atmosphere. The progress of the reaction was monitored by LC / MS. The crude reaction mixture contained starting material (compound 1) plus M ++H 693 (minor-1), 693 (major;3) and 477 (minor-2) showed three product peaks with similar retention times. After stirring the contents for 4 days (70% consumption), the reaction mixture was concentrated and diluted with water. The resulting pale yellow precipitate that formed was collected by filtration and dried. The crude solid was purified by chromatography on silica gel (10% NEt) with gradient elution from 70% EtOAc / Hexane to 100% EtOAc. 3 / CH 2 Cl 2 Compounds 1 and M were purified by column chromatography (pre-treatment with hexane and elution with hexane). + The fractions containing +H 693 were collected and concentrated. The resulting crude white solid was subjected to repurification in a similar manner as described above, except eluting with 30%-50%-75%-100% EtOAc / Hexanes. + The major product peak with +H 693 was collected as a white solid (270 mg, 18%) and characterized as N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3). 1 H NMR (DMSO-d6): δ 9.21(s,1H), 9.17(s,1H), 8.16(d,1H, J=2.6Hz), 7.76(d,1H, J=8.5Hz), 7.44(d,1H, J=8.5Hz), 7.02(s,2H), 5.78(d,1H, J 3 PH =6.1Hz), 3.64(s,6H), 3.58(s,3H), 1.45(s,6H), 1.33(s,9H).LCMS: Retention time: 14.70 minutes; Purity: 95%; MS(m / e): 693(MH + ). 31 P NMR(DMSO-d6):-11.36.

[0150] N4-(2,2-dimethyl-4-[(dihydrogen phosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (Compound 4) Trifluoroacetic acid (1.5 mL) was dissolved in CH under a nitrogen atmosphere at 0 °C. 2 Cl 2 N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3, 120 mg, 0.173 mmol) dissolved in 10 mL of ethyl acetate was added dropwise over a period of 5 minutes. The contents were allowed to stir for 1.5 hours. The progress of the reaction mixture was monitored by LC / MS. After complete consumption of the starting material, the reaction mixture was concentrated, dried and triturated with ether. The ether layer was decanted and dried to give a crude solid. LC / MS analysis of the crude product showed M + +H Three peaks were observed due to 581, 471 and 501. + The peak corresponding to +H 581 was collected by preparative HPLC chromatography purification. The fractions were lyophilized and dried to give 53 mg (52%) of an off-white fluffy solid, characterized as N4-(2,2-dimethyl-4-[(dihydrogenphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 4). 1 H NMR (DMSO-d6): δ 9.21(br s,2H), 8.16(d,1H, J=2.6Hz), 7.93(d,1H, J=8.5Hz), 7.39(d,1H, J=8.5Hz), 7.05(s,2H), 5.79(d,1H, J 3 PH =6.6Hz), 3.67(s,6H), 3.59(s,3H), 1.44(s,6H).LCMS: Retention time: 8.52 minutes; Purity: 95%; MS(m / e): 581(MH + ). 31 P NMR(DMSO-d6):-2.17.

[0151] Alternative synthesis of prodrug compound 4 An alternative method for synthesizing the prodrug compound 4 that alleviates the need for column chromatography and HPLC purification is presented below.

[0152] Synthesis of N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3) TIFF0007681570000029.tif137128N4-(2,2-dimethyl-3-oxo-4H-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 1, 19.73 g, 41.97 mmol) in DMF (100 mL), Cs 2 CO 3 (15.04 g, 46.16 mmol) and di-tert-butyl chloromethyl phosphate (13.0 g, 50.38 mmol) were stirred at room temperature under a nitrogen atmosphere. The progress of the reaction was monitored by in-process LC / MS. The crude reaction mixture contained starting material (compound 1) plus M + Two product peaks (ratio 1:6.5) with close retention times were observed, indicating +H 693 (minor) and 693 (major). The initial yellow reaction mixture turned olive green as the reaction proceeded. Workup was performed as follows: 1) After stirring the contents for 30 hours (92% consumption), the reaction mixture was poured onto ice water (400 mL) and the contents were stirred by adding a brine solution (200 mL). The fine tan solid that formed was filtered, washed with water, and dried overnight. 2) The solid (35 g) was dissolved in MTBE (500 mL) and washed with water (400 mL). The aqueous layer was extracted with MTBE (2×350 mL) until no UV was detected by TLC. The combined organic layers were washed with anhydrous Na 2 SO 4It was dried and decanted. NOTE: Step 2 can be carried out directly, however, DMF extraction back into solution leads to difficulties in the crystallization step. 3) The dark red, clear solution was treated with 10 g of activated charcoal, heated to boiling, and filtered. 4). The dark red clear solution was concentrated by normal heating to its volume of 400 mL and left for crystallization. The solid crystallized as granules was filtered, the granules were crushed into powder, washed with MTBE (400 mL) and dried under high vacuum. See step 7 for workup of mother liquor. Weight of solid: 17 g; Purity: 90% (compound 3), 6.26% (compound 1), 1.8% (minor M+ 693). 5) At this stage, the solid was taken up in 500 ml of ethyl ether and heated to boiling. It was cooled and filtered to remove undissolved material. The filtrate was concentrated. 6). The above concentrate was subjected to crystallization in MTBE (300 mL). The formed white solid was filtered, washed with MTBE (100 mL) and dried under high vacuum to obtain the desired N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3) with 97% purity. 1 H NMR (DMSO-d6): δ 9.21(s,1H), 9.17(s,1H), 8.16(d,1H, J=2.6Hz), 7.76(d,1H, J=8.5Hz), 7.44(d,1H, J=8.5Hz), 7.02(s,2H), 5.78(d,1H, J 3 PH =6.1Hz), 3.64(s,6H), 3.58(s,3H), 1.45(s,6H), 1.33(s,9H).LCMS: Retention time: 14.70 minutes; Purity: 95%; MS(m / e): 693(MH + ). 31 P NMR (DMSO-d6): -11.36. Weight of solid: 15.64 g (yield: 55%); Purity: 97% (R935787), 3% (compound 1). 7) The mother liquor was concentrated and steps 5 and 6 were repeated to obtain compound 3.

[0153] Synthesis of N4-(2,2-dimethyl-4-[(dihydrogen phosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 4) AcOH:H 2 N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 3); (15.0 g, 21.67 mmol) dissolved in 2,4-dimethyl-3,4-dimethyl-2 ...

[0154] The reaction mixture was cooled, poured onto ice water (200 mL), stirred for 20 min, and filtered. The clear white filter cake was washed successively with water (600 ml) and acetone (200 mL), dried for 2 h, and then placed in a desiccator for 30 min. 2 O 5 and dried under high vacuum. Weight of solid: 12.70 g; Purity: 97% (compound 3) and 3% (compound 1). 1 1 H NMR indicated the presence of acetic acid (1:1).

[0155] To remove acetic acid, the solid was taken up in acetonitrile (300 mL) and concentrated by rotovap vacuum. This process was repeated twice with acetonitrile and toluene (3×300 mL). The resulting solid was dried under high vacuum at 50° C.

[0156] Finally, the solid was taken up in acetone (400 mL), filtered and dried to give N-(2,2-dimethyl-4-[(dihydrogenphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 4). 1 H NMR (DMSO-d6): δ 9.21(br s,2H), 8.16(d,1H, J=2.6Hz), 7.93(d,1H, J=8.5Hz), 7.39(d,1H, J=8.5Hz), 7.05(s,2H), 5.79(d,1H, J 3 PH =6.6Hz), 3.67(s,6H), 3.59(s,3H), 1.44(s,6H).LCMS: Retention time: 8.52 minutes; Purity: 95%; MS(m / e): 581(MH + ). 31 P NMR(DMSO-d6): -2.17.

[0157] Synthesis of N4-(2,2-dimethyl-4-[(dihydrogen phosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine monocalcium salt (compound 6) TIFF0007681570000030.tif69142NaHCO 3 (0.17 g, 2.02 mmol) in water (10 mL) was added dropwise to a suspension of N4-(2,2-dimethyl-4-[(dihydrogenphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (0.5 g, 0.86 mmol) in water (5 mL) at room temperature while stirring the contents. The formed clear solution was diluted with CaCl in dropwise form at room temperature. 2The mixture was treated with an aqueous solution (10 mL) (0.11 g, 0.99 mmol in 10 mL water). The addition resulted in the precipitation of a white solid from the reaction mixture. After complete addition, the contents were stirred for a period of 30 minutes, filtered, washed with water (40 mL), and dried. The clear white solid was taken up in water (30 mL) and heated to boiling on a stir plate. The solution was cooled, filtered, and dried. The white solid was collected and further dried under high vacuum at 80° C. for 32 hours to give 0.41 g (83%) of N4-(2,2-dimethyl-4-[(dihydrogenphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine monocalcium salt (compound 6).

[0158] Synthesis of prodrug compound 8 TIFF0007681570000031.tif34141N4-(2,2-dimethyl-4-[(di-tert-butylphosphonooxy)methyl]-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (prepared as described above) (0.2 g, 0.29 mmol) was dissolved in MeOH (5 mL) and Et 2 A mixture of 2,2-dimethyl-4-methoxymethyl-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 8) was added to the mixture in 5 mL of 1H2O. 2N aqueous NaOH (0.023 g, 0.58 mmol) was added in one portion while stirring the contents at room temperature. The progress of the reaction was monitored by LC / MS. After stirring for 8 h, the precipitated solid was filtered and dried to give N4-(2,2-dimethyl-4-methoxymethyl-3-oxo-5-pyrido[1,4]oxazin-6-yl)-5-fluoro-N2-(3,4,5-trimethoxyphenyl)-2,4-pyrimidinediamine (compound 8) as a white solid (0.11 g, 74%). 1H NMR(DMSO-d6):δ 9.47(s,1H), 9.15(s,1H), 8.16(d,1H, J=3.8Hz), 7.87(d,1H, J=8.5Hz), 7.37(d,1H, J=8.5Hz), 7.03(s,2H), 5 .40(s,2H), 3.66(s,6H), 3.59(s,3H), 3.27(s,3H), 1.44(s,6H).LCMS: Retention time: 12.88 minutes; Purity: 92%; MS(m / e): 515(MH) + ).

[0159] Example 5 In vitro testing of compounds for efficacy in treating CRS material 1. Human PBMC cells (PPA Research Group, Cat No. 15-00021) 2. RPMI medium 10% FBS 3. GMCSF (Peprotech, Cat No. 300-03) and IL4 (Peprotech Cat No. 200-04) 4. White clear bottom 96 well plate (Fisher, Cat No. 07-200-587, Corning #3903) 5. Human IL-2 DuoSet ELISA (R&D Systems, Cat No. DY202) 6. Human IL-6 DuoSet ELISA (R&D Systems, Cat No. DY206) 7. Cell Titer Glo Reagent (Promega, Cat No. G7573) 8. Dynabeads Human T-Activator CD3 / CD28 (Fisher, Cat No. 111.61D) 9. Anti-human CD3 (BD ​​Biosciences, Cat No. 555336) 10. CD28, clone CD28.2 (Beckman Coulter Inc. Cat No. IM1376) 11. Recombinant human IL-2 protein (R&D Systems, Cat No. 202-IL-500)

[0160] DC Differentiation: Human PBMC cells (400 million) obtained from a supplier are transferred to three T-175 flasks containing 16 ml of RPMI medium (10% FBS) and incubated at 37° C. for 2 hours. After 2 hours, floating PBLs are removed and the cells are rinsed twice with 10 ml of medium. PBLs and medium are reserved for T cell expansion. 16 ml of fresh RPMI medium (10% FBS) containing GMCSF (100 ng / ml) and IL4 (20 ng / ml) are added and the flasks are kept in a 37° C. incubator. After 3 days, fresh GMCSF (100 ng / ml) and IL4 (20 ng / ml) are added to the flasks and incubation is continued.

[0161] T cell proliferation: 1. Coat a T-175 flask with 16ml of PBS containing 1ug / ml anti-CD3 (16ul of 1mg / ml stock) and 5ug / ml anti-cd28 (400ul of 200ug / ml stock) for approximately 2 hours. 2. Settling, 2 x E 8 of PBLs are resuspended in 60 ml of RPMI medium (10% FBS) containing 60 μl of IL2. 3. The coating solution is aspirated from the flask and the cells are added to the stimulation flask. 4. After 3 days, tap stimulation flask to dislodge cells stuck to the bottom of the flask. Re-seed a new T-175 at 1xE6 cells / ml in 60ml media containing 60μl IL2.

[0162] CRS Assay: After 1.4 days, dendritic cells are harvested by sedimentation (1000 rpm / 10 min) and aspirating the medium. Resuspend cells in fresh RPMI medium (10% FBS) and plate cells (25K / well in 50 μl) onto white clear bottom 96-well plates. 2. 100 μl of RPMI medium containing 2x concentrated test compound per well is added to the above cell culture medium (final concentration is 1x) and the plate is pre-incubated at 37° C. for 1 hour. 3. After 1 hour compound pre-incubation, add 50 μl T cells per well (1.7k / well) along with CD3 / CD28 beads (1.7k / well). Plates are incubated overnight at 37° C. 4. Collect 80 μl of supernatant from each well for IL6 ELISA and 80 μl for IL2 ELISA. ELISA is performed according to the instructions from R&D Systems. Add 25 μl of Cell Titer Glo reagent to the remaining 40 μl / well of the cell culture plate, incubate on a shaker for 1-2 minutes, and read the plate for luminescence intensity to determine compound cytotoxicity.

[0163] TIFF0007681570000032.tif35142NB: IL-6 is produced primarily by dendritic cells activated by T cells, and IL-2 is produced exclusively by activated T cells.

[0164] In view of the many possible embodiments to which the principles of the disclosed invention may be applied, it should be recognized that the illustrated embodiments are merely preferred examples of the invention and should not be construed as limiting the scope of the invention, which is rather defined by the following claims, and we therefore claim as our invention all that comes within the scope and spirit of these claims.

Claims

1. A pharmaceutical composition for use in a method for treating and / or preventing cytokine release syndrome (CRS) in a subject suffering from or at risk of developing CRS, comprising a compound of the formula: or a salt, solvate, or N-oxide thereof, wherein R ３０ is H or phosphonooxyalkyl.

2. The compound is or a salt and / or solvate thereof.

3. The compound is The pharmaceutical composition according to claim 1 or 2,

4. The compound is The pharmaceutical composition of claim 1 ,

5. The pharmaceutical composition of any one of claims 1 to 4, which ameliorates a sign or symptom of CRS compared to the severity of the sign or symptom prior to administration of the pharmaceutical composition.

6. The pharmaceutical composition of claim 5 , wherein the sign or symptom is fever.

7. The object is The subject has previously been treated with a first therapy for which CRS is a known, possible, or potential side effect; or are about to receive or are currently receiving a first treatment for which CRS is a known, possible, or potential side effect; The pharmaceutical composition according to any one of claims 1 to 6.

8. The pharmaceutical composition of claim 7 , wherein the first treatment comprises a cell therapy.

9. 9. The pharmaceutical composition of claim 8, wherein the cell therapy comprises chimeric antigen receptor (CAR) expression therapy, transgenic receptor therapy, or a combination thereof.

10. The pharmaceutical composition of any one of claims 1 to 9, administered in combination with a second therapeutic agent.

11. 11. The pharmaceutical composition of claim 10, wherein the second therapeutic agent is a steroid, an anti-inflammatory agent, an immunosuppressant, or a combination thereof.

12. The steroid may be alclometasone, algestone, beclomethasone, betamethasone, budesonide, clobetasol, clobetasone, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difluprednate, enoxolone, fluazacort, flucloronide, fludrocortisone, flumethasone, flunisolide, fluocinolone, fluocinonide, fluocortin, fluocortolone, fluorometh ... bromine, fluperolone, fluprednidene, fluprednisolone, flurandrenolide, fluticasone, formocortal, halcinonide, halobetasol, halometasone, halopredone, hydrocortamate, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone, paramethasone, prednicarbate, prednisolone, prednisone, prednibal, prednylidene, rimexolone, tixocortol, triamcinolone, or any combination thereof; the anti-inflammatory agent is an aminosalicylate, a cyclooxygenase inhibitor, diclofenac, etodolac, famotidine, fenoprofen, flurbiprofen, ketoprofen, ketorolac, ibuprofen, indomethacin, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, piroxicam, salsalate, sulindac, tolmetin, or a combination thereof; or 12. The pharmaceutical composition of claim 11, wherein the immunosuppressant is mercaptopurine, a corticosteroid, an alkylating agent, a calcineurin inhibitor, an inhibitor of inosine monophosphate dehydrogenase (IMPDH), an agent designed to preserve the recipient's humoral immune response intact while suppressing cellular immunity, or a combination thereof.

13. 12. The pharmaceutical composition of claim 11, wherein the second therapeutic agent is dexamethasone or prednisone, or a combination thereof.

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