USOO892.1361 B2

(12) United States Patent (10) Patent No.: US 8,921,361 B2 Cmiljanovic et al. (45) Date of Patent: Dec. 30, 2014

(54) TRIAZINE, PYRIMIDINE AND PYRIDINE 409/04 (2013.01); C07D 413/04 (2013.01); ANALOGS AND THEIR USEAS C07D 413/14 (2013.01); C07D 417/04 THERAPEUTICAGENTS AND DAGNOSTIC (2013.01); C07D 417/14 (2013.01); C07D PROBES 491/048 (2013.01); C07D491/147 (2013.01); C07D 495/04 (2013.01); C07D 495/14 (2013.01); C07D498/04 (2013.01); C07D (75) Inventors: Vladimir Cmiljanovic, Basel (CH): 513/04 (2013.01); C07D 519/00 (2013.01) Natasa Cmiljanovic, Basel (CH); Bernd USPC ...... 514/232.2:544/83 Giese, Fribourg (CH); Matthias (58) Field of Classification Search Wymann, Bern (CH) None See application file for complete search history. (73) Assignee: University of Basel, Basel (CH) (56) References Cited (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 U.S. PATENT DOCUMENTS U.S.C. 154(b) by 60 days. 5,489,591 A * 2/1996 Kobayashi et al...... 514,245 (21) Appl. No.: 13/128,436 7,173,029 B2 2/2007 Hayakawa et al. 8,217,036 B2 * 7/2012 Venkatesan et al...... 514,232.2 (22) PCT Filed: Nov. 10, 2009 2010 OO69629 A1 3/2010 Shimma et al. (86). PCT No.: PCT/B2O09/OOT404 FOREIGN PATENT DOCUMENTS EP 1864 665 A1 12/2007 S371 (c)(1), WO 2005/028444 A1 3, 2005 (2), (4) Date: Jul. 1, 2011 WO 2007 1271.75 A2 11/2007 WO 2008/O18426 A1 2, 2008 (87) PCT Pub. No.: WO2010/052569 WO 2009/093981 A1 T 2009 WO 2009/143313 A1 11, 2009 PCT Pub. Date: May 14, 2010 WO 2009/143317 A1 11, 2009 (65) Prior Publication Data OTHER PUBLICATIONS US 2011/0275762 A1 Nov. 10, 2011 Menicagli R. et al: “2-Alkyl-4,6-dialkylamino-1,3,5-triazines via Grignard Alkylation of Cyanuric Chloride: An Aged Reaction Revis (30) Foreign Application Priority Data ited' in: Tetrahedron, Elsevier Science Publishers, vol. 56, No. 49, Dec. 1, 2000, pp. 9705-9711. Nov. 10, 2008 (GB) ...... O821219.3 Matsuno T et al: “Synthesis and Antitumor Activity of Benzimidazolyl-1,3,5-Triazine and Benzimidazolylpyrimidine (51) Int. Cl. Derivatives' in: Chemical and Pharmaceutical Bulletin, Pharmaceu CO7D40 L/04 (2006.01) tical Society of Japan, vol. 48, No. 11, Nov. 1, 2000, pp. 1778-1781. CO7D 403/04 (2006.01) A 6LX3/5.375 (2006.01) * cited by examiner CO7D 2.5L/8 (2006.01) CO7D 40/12 (2006.01) Primary Examiner — Alicia L Otton CO7D 403/2 (2006.01) (74) Attorney, Agent, or Firm — Joyce Von Natzmer, Agris CO7D 403/4 (2006.01) & von Natzmer LLP CO7D 405/04 (2006.01) CO7D 405/4 (2006.01) (57) ABSTRACT CO7D 409/04 (2006.01) The invention relates to novel therapeutic agents and diag CO7D 413/04 (2006.01) nostic probes. The invention also relates to phosphoinositide CO7D 413/4 (2006.01) 3-kinase (PI3K) and mammalian target of rapamycin CO7D 417/04 (2006.01) (mTOR) inhibitor triazine-, pyrimidine- and pyridine-based CO7D 417/4 (2006.01) compounds Formula (I), their stereoisomers, geometric iso CO7D 49/048 (2006.01) mers, tautomers, Solvates, metabolites, N-oxide derivatives, CO7D 49/47 (2006.01) pharmaceutically acceptable salts, and prodrugs thereof.com CO7D 495/04 (2006.01) positions of the new compounds; either alone or in combina CO7D 495/4 (2006.01) tion with at least one additional therapeutic agent, with a CO7D 49.8/04 (2006.01) pharmaceutically acceptable carrier; and uses of the new CO7D 53/04 (2006.01) compounds, either alone or in combination with at least one C07D 519/00 (2006.01) additional therapeutic agent, for treating disorders mediated (52) U.S. Cl. by lipid kinases. Methods of using compounds of Formula CPC ...... C07D401/04 (2013.01); C07D 251/18 (I) for in vitro, in situ, and in vivo diagnosis, prevention or (2013.01); C07D401/12 (2013.01); C07D treatment of such disorders in mammalian cells, or associated 403/04 (2013.01); C07D 403/12 (2013.01); pathological conditions, are disclosed. (Formula I) C07D 403/14 (2013.01); C07D405/04 (2013.01); C07D405/14 (2013.01); C07D 16 Claims, No Drawings US 8,921,361 B2 1. 2 TRIAZINE, PYRIMIDINE AND PYRIDINE years, the connection between cancer and PI3K has been ANALOGS AND THEIR USEAS further substantiated Cully et al., Nat. Rev., Cancer 6:184 THERAPEUTICAGENTS AND DAGNOSTIC 192 (2006); Wymannet al., Curr. Opin. Cell Biol. 17:141-149 PROBES (2005); Vivanco et al., Nat. Rev., Cancer 2:489-501 (2002). PI3Ks have since been recognized to modulate a wide range This is the U.S. national stage of International application of cellular activities, and to be central to the growth and PCT/IB2009/007.404, filed Nov. 10, 2009 designating the metabolic control. Genetically modified mice targeting the United States and claims priority to GB 0821219.3, filed Nov. PI3K pathway, and the elucidation of human hereditary dis 10, 2008. ease like Cowden's syndrome, tuberous Sclerosis, ataxia 10 telangiectasia, X-linked myotubular myopathy and Charcot FIELD OF THE INVENTION Marie-Tooth neuropathy, have provided further insight in the cellular and systemic role of phosphoinositide signaling. The invention relates to new therapeutic agents and diag Deregulation of phosphoinositide levels, and in particular the nostic probes, including pharmaceutically acceptable salts, product of class I PI3Ks, PtdIns(3,4,5)P3, is involved in the prodrugs and metabolites thereof, which are useful for modu 15 pathogenesis of cancer, chronic inflammation, allergy, meta lating protein or enzyme activity for modulating cellular bolic disease, diabetes and cardiovascular problems. activities such as signal transduction, proliferation, differen PI3Ks are a family of enzymes, which phosphorylate the tiation, programmed cell death, migration and cytokine secre 3'-OH position of the inositol ring of phosphoinositides. They tion. More specifically the invention provides compounds have been divided into three classes on the basis of structural which inhibit, regulate, detect and/or modulate kinase activ features and in vitro lipid substrate specificity (Marone et al. ity, in particular phosphoinositide 3-kinase (PI3K), mamma Biochimica et Biophysica Acta 1784:159-185 (2008). Class lian target of rapamycin (mTOR), DNA-PK and ATM kinase I PI3Ks form heterodimers, which consist of one of the four inhibitor compounds, their pharmaceutically acceptable closely related ~110kDa catalytic Subunits, and an associated salts, and prodrugs thereof compositions of the new com regulatory Subunit belonging to two distinct families. In vitro pounds, either alone or in combination with at least one 25 they are capable to convert PtdIns to PtdIns-3-P, PtdIns-4-P to additional therapeutic agent, with a pharmaceutically accept PtdIns(3,4)P2, and PtdIns(4,5)P2 to PtdIns(3,4,5)P3, but the able carrier, and uses of the new compounds, either alone or in vivo substrate is PtdIns(4,5)P2 Cantley et al., Science in combination with at least one additional therapeutic agent, 2.96:1655-1657 (2002). Class I PI3Ks are activated by a large in the prophylaxis or treatment of a number of diseases, in variety of cell-surface receptors, comprising growth factor particular, those characterized by the abnormal activity of 30 receptors as well as G protein-coupled receptors. serine/threonine kinases, receptor tyrosine kinases and lipid Class II PI3Ks are capable to phosphorylate PtdIns and kinases. The invention also relates to methods of using the PtdIns-4-P in vitro, but their relevant in vivo substrates are compounds for in vitro, in situ, and in vivo diagnosis, assay still under investigation. This class of large (170-200 kDa) development or treatment of mammalian cells, or associated enzymes has three members, all characterized by a C-termi pathological conditions. 35 nal C2 homology domain. No adaptor molecules for class II PI3Ks have been identified so far. Class III PI3Ks are solely BACKGROUND OF THE INVENTION able to phosphorylate PtdIns, and thus generate only PtdIns 3-P. The single member of this class is Vps34, of which the S. Protein kinases participate in the signaling events which cerevisiae Vps34p (vacuolar protein Sorting mutant 34 pro control the activation, growth, differentiation, Survival and 40 tein) is the prototype, and has been shown to play an essential migration of cells in response to extracellular mediators or role in trafficking of newly synthesized proteins from the stimuli including growth factors, cytokines or chemokines. In Golgi to the yeast vacuole, an organelle equivalent to lysos general, these kinases are classified in two groups, those that omes in mammals Schu et al., Science 260:88-91 (1993). preferentially phosphorylate tyrosine residues and those that Phosphoinositide 4-kinases (PI4Ks) phosphorylate the preferentially phosphorylate serine and/or threonine resi 45 4'-OH position of the inositol ring of PtdIns, and thereby dues. The tyrosine kinases include membrane-spanning generate PtdIns-4-P. This lipid can then be further phospho growth factor receptors, for example the epidermal growth rylated by PtdIns-4-P 5-kinases to generate PtdIns(4,5)P2. factor receptor (EGFR) and cytosolic non-receptor kinases which is the main source for phospholipase C and PI3K including Src family kinases, the Syk family kinases and the signaling at the plasma membrane. Four PI4Ks isoforms are Tec family kinases. 50 known: PI4KIIC. and B, and PI4KIIIC. and B. The PI4KIIIs are Inappropriately high protein kinase activity is involved in most closely related to PI3Ks. many diseases including cancer, metabolic diseases, immu The class of PI3K-related proteins, referred to as class IV nological diseases and inflammatory disorders. This can be PI3Ks, consists of high molecular weight enzymes with a caused either directly or indirectly by the failure of control catalytic core similar to PI3Ks and PI4Ks and include the mechanisms due to mutation, overexpression or inappropri 55 target of rapamycin (mTOR, also known as FRAP), DNA ate activation of the enzyme. dependent protein kinase (DNA-PKcs), the ataxia telang Protein tyrosine kinases—both receptor tyrosine kinases iectasia mutated gene product (ATM), ataxia telangiecta and non-receptorkinases—are essential for the activation and siarelated (ATR), SMG-1 and transformation/transcription proliferation of cells of the immune system. Among the ear domain-associated protein (TRRAP). The first five members liest detectable events upon the immunoreceptor activation in 60 are active protein serine-threonine kinases that are involved in mast cells, T cells and B cells is the stimulation of non cell growth control and genome/transcriptome Surveillance receptor tyrosine kinases. (Marone et al., Biochimica et Biophysica Acta 1784:159-185 Phosphoinositide 3-kinases (PI3Ks) were early on identi (2008). DNA-PKcs, ATM, ATR and SMG-1 are involved in fied as lipid kinases associated with viral oncogens Whitman DNA-damage responses. The only active kinase not involved et al., Nature 315:239-242 (1985): Sugimoto et al., Proc. 65 in DNA-damage is mTOR, which is regulated by growth Natl. Acad. Sci. 81:21.17-2121 (1984); Macara et al., Proc. factors and nutrient availability, and coordinates protein Syn Natl. Acad. Sci. 81:2728-2732 (1984), and for the last 20 thesis, cell growth and proliferation. Target of rapamycin US 8,921,361 B2 3 4 (mTOR) complexes 1 and integrate growth factor signaling known, which has been developed as analogue of antitumor (via PI3K/PKB and the Ras/MAPK cascade), energy status agent triethylenemelamine (TEM); HMM acts as a prodrug of (LKB1 and AMPK) and nutrient detection. TOR is positively hydroxymethylpentamethylmelamine (HMPMM: metaboli regulated by PKB/Akt, which phosphorylates the negative cally active type of HMM) Johnson et al., Cancer, 42:2157 regulator TSC2 in the tuberous sclerosis complex (TSC), 2161 (1978). HMM has been marketed in Europe under the resulting in activation of the GTPase Rheb and mTOR (Ma indications for the treatment of ovarian and Small cell lung rone et al., Biochimica et Biophysica Acta 1784:159-185 CaCCS. (2008). In parallel. mTOR stimulates translation of riboso Certain triazine compounds are known to have PI3 kinase mal proteins and therefore ribosome biogenesis via the acti inhibitor activity and inhibit the growth of cancer cells WO02088112 (EP1389617), “HETEROCYCLIC COM vation of p70' Wullschleger et al., Cell 124:471 (2006). 10 POUNDS AND ANTITUMOR AGENT CONTAINING Rapamycin, and its derivatives RAD001 and CCI-779, bind THE SAMEAS ACTIVE INGREDIENT, Kawashima et al., to FKBP12, and the complex blocks mTOR complex 1 Filing date: 26 Apr. 2002: WO05095389 (EP1741714), (mTORC1) activity very selectively. Various clinical trials HETEROCYCLIC COMPOUND AND ANTI-MALIG were initiated using Rapamycin and derivatives, mostly in NANT-TUMORAGENT CONTAINING THE SAME AS patients with tumors displaying elevated PI3K signaling and 15 ACTIVE INGREDIENT', Kawashima et al., Filing date: 30 hyperactive mTOR. Promising results were obtained in Mar 2005; WO06095906 (EP1864665), “IMMUNOSUP mantle cell lymphoma, endometrial cancer and renal cell PRESSIVE AGENT AND ANTI-TUMORAGENT COM carcinoma Guertin et al., Cancer Cell 12:9 (2007). Rapa PRISING HETEROCYCLIC COMPOUND AS ACTIVE mycin and its derivatives possess anti-angiogenic activity INGREDIENTS, Haruta et al., Filing date: 11 Mar. 2005; because they counteract VEGF action Guba et al., Nat. Med. WO09905138 (EP1020462), HETEROCYCLIC COM 8:128 (2002). This opens avenues for combinatorial treat POUNDS AND ANTITUMOR AGENT CONTAINING ments with conventional chemotherapy Beuvink et al., Cell THE SAME AS ACTIVE INGREDIENT, Kawashima et al., 120:747 (2005). Filing date: 24 Jul.1998;). The triazine compound ZSTK474, The PI3K pathway is a key signaling transduction cascade developed in research laboratories of Zenyaku Kogyo is the controlling the regulation of cell growth, proliferation, Sur 25 first orally administered triazine compound highly active vival as well as cell migration. PI3Ks are activated by a wide against PI3KS that displayed potent antitumor activity against variety of different stimuli including growth factors, inflam human cancer Xenografts in mice, without evidence of critical toxicity Yaguchi et al., Journal of the National Cancer Insti matory mediators, hormones, neurotransmitters, and immu tute, 98:545-556, (2006). ZSTK474 is an ATP-competitive noglobulins and antigens Wymann et al., Trends Pharmacol. inhibitor of class I phosphatidylinositol 3-kinase isoforms Sci. 24:366-376 (2003). The class IAPI3K isoforms PI3Kc, 30 Kong et al., Cancer Sci., 98: 1638-1642 (2007). B and 8, are all bound to one of the p85/p55/p50 regulatory Certain pyrimidine compounds are known to have p110 subunits, which all harbor two SH2 domains that bind with alpha binding, PI3 kinase inhibitor activity and inhibit the high affinity to phosphorylated Tyr-X-X-Met motifs. These growth of cancer cells IP of AstraZeneca: WO07066103, motifs are present in activated growth factor receptors, their WO07080382, WO08023159, WO08023180, WO08032027, Substrates and numerous adaptor proteins. As described 35 WO08032033, WO08032036, WO08032041, WO08032072, above, activation of the PI3K/PKB signaling cascade has a WO08032077, WO08032086, WO08032089, positive effect on cell growth, survival and proliferation. Con WO08032091; IP of Genentech/Piramed/Roche: stitutive up-regulation of PI3K signaling can have a deleteri US200700.9880, WO07127183. WO08073785, ous effect on cells leading to uncontrolled proliferation, WO07042810, WO07122410, WO071271 75, WO07129161, enhanced migration and adhesion-independent growth. 40 WO08070740, WO2006046031, WO2006046040, These events favor not only the formation of malignant WO2007042806, WO2007122410; IP of Novartis: tumors, but also the development of inflammatory and WO07084786, WO08098058. autoimmune disease. In order to expand antitumor spectrum of and increase The PI3 kinase/Akt/PTEN pathway is an attractive target antitumor activities of such compounds, active against PI3KS for cancer drug development since Such agents would be 45 and/or mTOR, the inventors carried out intensive studies on expected to inhibit proliferation, reverse the repression of triazine-, pyrimidine- and pyridine-based derivatives. They apoptosis and Surmount resistance to cytotoxic agente in can thus prepared new heterocyclic compounds represented by cer cells. PI3 kinase inhibitors have been reported see nota the formula (I) and formulas (Ia) to (Ii) which exhibit strong bly Marone et al., Biochimica et Biophysica Acta 1784:159 biological activity against lipid kinases. 185 (2008); Yaguchi et al. (2006) Jour. Of the Nat. Cancer 50 In comparison with the PI3K inhibitors disclosed by Zeny Inst. 98(8):545-556; U.S. Pat. No. 7,173,029; U.S. Pat. No. aku Kogyo WO02088112 (EP1389617), WO2005095389 7,037,915; U.S. Pat. No. 6,608,056; U.S. Pat. No. 6,608,053; (EP 1741714), WO2OO6095906 (EP1864665), U.S. Pat. No. 6,838,457; U.S. Pat. No. 6,770,641; U.S. Pat. WO09905138 (EP1020462), AstraZeneca (WO07066103, No. 6,653,320; U.S. Pat. No. 6,403,588; U.S. Pat. No. 6,703, WO07080382, WO08023159, WO08023180, WO08032027, 414; WO9715658; WO2006046031: WO2006046035: 55 WO08032033, WO08032036, WO08032041, WO08032072, WO2006046040; WO2007042806; WO2007042810; WO08032077, WO08032086, WO08032089, WO2004017950. US2004092561; WO2004007491; WO08032091), Piramed/Genentech (US2007009880, WO2004006916: WO2003037886; US2003149074; WO07127183, WO08073785, WO07042810, WO07122410, WO2003035618: WO2003034997; WO2007084786; WO07127175, WO08070740, WO2006046031, WO2007095588; WO2008098058; US2003158212: 60 WO2006046040, WO2007122410), Yamanouchi/Piramed EP1417976; US2004053946; JP2001 247477; JP08175990; (WO01083456) and Novartis (WO07084786, JP08176070). WO08098058), the inhibitors of the invention differ in the 1,3,5-triazine and pyrimidine derivatives as pharmaceuti insertion of a Natom in the basic heterocyclic ring that makes cals have been made with respect to antitumor, anti-inflam better biological activity to the target enzyme, and/or in the matory, analgesic and antispasmodic activities. Especially, 65 insertion of a novel molecular fragment making the whole hexamethylmelamine or altretamin (HMM or N,N',N',N', molecule more active or more selective to the appropriate N,N'-hexamethyl-1,3,5-triazine-2,4,6-triamine) is well enzyme. US 8,921,361 B2 5 6 SUMMARY OF THE INVENTION -continued (Ic) The invention relates generally to new triazine-, pyrimi dine- and pyridine-based derivatives and their use as thera peutic agents and diagnostic probes. r The invention also relates to kinase inhibitors and kinase diagnostic probes. R3, 1. R3 The invention also relates to phosphoinositide 3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibi 10 R" l alss tor compounds with anti-cancer activity, pharmaceutical for Ny. N R2 mulations thereof, which are potentially useful intreatment of Triazine disease, conditions and/or disorders modulated by PI3K and mTOR kinases. The compounds may inhibit tumor growth in (Id) mammals and may be useful for treating human cancer 15 patients. The invention also relates to methods of using the com R3.DCC R3, pounds for in vitro, in situ, and in vivo diagnosis or treatment R3, 1. R3 of mammalian cells, organisms, or associated pathological conditions. Specifically, one aspect of the invention provides com l 2 pounds of formula W. R N Sw More specifically, one aspect of the invention provides 25 Rs N N triazine compounds of formulas (Ia) to (Id), pyrimidine com H pounds of formulas (If) to (Ii) and pyridine compounds of formulas (Ie) and (If) to (Ili): Triazine (Ie) 30 (I) R3.DC X DC R3, R3.DC X I R3, R3, 1. R3 R3, 1. R3 35 o1 No o1 No R" n Y.- Uals R2

R ls U als R2 Pyridine (U=N; Q, G = C) 40 Pyridine (G = N: Q, U = C) (Ia) Pyridine (Q = N: U, G = C) R3.DC X DC R3, (If) R3, 1. R3 45 r C R3, R3, N R3

s Y, n G R us Nals R2 50 Rs N els Triazine Z 4. R (Ib) R3, Fused Pyrimidine (G = N) Fused Pyridine (G = C)

55 (Ig) R3, 1. R R N1 SN R3.DC C R3, R4. l als R3, N R3 N N R 60 Z O Rs -{ No. R4 \ Nals R R47 65 Fused Pyrimidine (G = N) Triazine Fused Pyridine (G = C) US 8,921,361 B2 8 -continued duced platelet aggregation, chronic myelogenous leukaemia (Ih) (CML), liver disease, pathologic immune conditions involv ing T cell activation, and CNS disorders. Another aspect of the invention provides methods of pre X DC R3, venting or treating a hyperproliferative disorder, comprising R3, N R3 administering to a mammal in need of Such treatment an effective amount of a compound of formula (I) or one of Y. n G formulas (Ia) to (Ii), or a stereoisomer, geometric isomer, tautomer, solvate, metabolite, N-oxide derivative or pharma ceutically acceptable salt or prodrug thereof, alone or in com 10 bination with one or more additional compounds having anti N R2 hyperproliferative properties. R’Kera. In a further aspect the present invention provides a method Fused Pyrimidine (G = N) of using a compound of this invention to treat a disease or Fused Pyridine (G = C) condition modulated by PI3 kinase and/or mTOR in a mam (II) 15 mal. R3. X R3, An additional aspect of the invention is the use of a com pound of this invention in the preparation of a medicament for the treatment or prevention of a disease or condition modu R3, N R3 lated by PI3 kinase in a mammal. R9 SC ra Another aspect of the invention includes kits comprising a compound of formula (I) or one of formulas (Ia) to (Ii), or a Stereoisomer, geometric isomer, tautomer, Solvate, metabo \ / r lite, N-oxide derivative orpharmaceutically acceptable salt or Y: Nals R2 prodrug thereof, a container, and optionally a package insert 25 or label indicating a treatment. Fused Pyrimidine (G = N) Another aspect of the invention includes methods of pre Fused Pyridine (G = C) paring, methods of separating, and methods of purifying compounds of formula (I) or one of formulas (Ia) to (Ii). Another aspect of the invention includes novel intermedi and stereoisomers, geometric isomers, tautomers, Solvates, ates useful for preparing formula (I) or one of formulas (Ia) to metabolites, N-oxide derivatives and pharmaceutically 30 (Ii). acceptable salts thereof. Additional advantages and novel features of this invention Another aspect of the invention provides a pharmaceutical shall be set forth in part in the description that follows, and in composition comprising a triazine or a pyrimidine or a pyri part will become apparent to those skilled in the art upon dine compound of formula (I) or one of formulas (Ia) to (Ii) examination of the following specification or may be learned and a pharmaceutically acceptable carrier. The pharmaceuti 35 by the practice of the invention. The advantages of the inven cal composition may further comprise one or more additional tion may be realized and attained by means of the instrumen therapeutic agents selected from anti-proliferative agents, talities, combinations, compositions, and methods particu anti-inflammatory agents, immunomodulatory agents, neuro larly pointed out in the appended claims. tropic factors, agents for treating blood disorders, agents for 40 DETAILED DESCRIPTION OF EXEMPLARY treating diabetes, and agents for treating immunodeficiency EMBODIMENTS disorders. Another aspect of the invention provides methods of inhib Reference will now be made in detail to certain embodi iting PI3 kinase activity, comprising contacting a PI3 kinase ments of the invention, examples of which are illustrated in with an effective inhibitory amount of a compound of formula 45 the accompanying structures and formulas. While the inven (I) or one of formulas (Ia) to (Ii), or a stereoisomer, geometric tion will be described in conjunction with the enumerated isomer, tautomer, solvate, metabolite, N-oxide derivative or embodiments, it will be understood that they are not intended pharmaceutically acceptable salt or prodrug thereof. to limit the invention to those embodiments. On the contrary, Another aspect of the invention provides methods of pre the invention is intended to cover all alternatives, modifica venting or treating a disease or disorder modulated by PI3 50 tions, and equivalents which may be included within the kinases, comprising administering to a mammal in need of scope of the present invention as defined by the claims. One Such treatment an effective amount of a compound of formula skilled in the art will recognize many methods and materials (I) or one of formulas (Ia) to (Ii), or a stereoisomer, geometric similar to equivalent to those described herein, which could isomer, tautomer, solvate, metabolite, N-oxide derivative or be used in the practice of the present invention. The present pharmaceutically acceptable salt or prodrug thereof. 55 invention is in no way limited to the methods and materials Examples of Such diseases, conditions and disorders include, herein described. but are not limited to, hyperproliferative disorders (e.g., can In the event that one or more of the literature, patents, and cer, including melanoma and other cancers of the skin), neu similar materials referred to differs from or contradicts the rodegeneration, cardiac hypertrophy, pain, migraine, neu present application, as regards including but not limited to rotraumatic diseases, stroke, diabetes, hepatomegaly, 60 defined terms, term usage, described techniques, or the like, cardiovascular disease, Alzheimer's disease, cystic fibrosis, only the teaching of the present application will be taken into autoimmune diseases, atherosclerosis, restenosis, psoriasis, consideration. allergis disorders, inflammation, neurological disorders, hor mone-related diseases, conditions associated with organ DEFINITIONS transplantation, immunodeficiency disorders, destructive 65 bone disorders, hyperproliferative disorders, infectious dis The term “alkyl as used herein refers to a saturated linear eases, conditions associated with cell death, thrombin-in or branched-chain monovalent hydrocarbon radical of one to US 8,921,361 B2 9 10 twelve carbonatoms (C1-C12), wherein the alkyl radical may groups include, but are not limited to, radicals derived from be optionally substituted independently with one or more benzene(phenyl), Substituted benzenes, naphthalene, substituents described below. In another embodiment, an anthracene, biphenyl, indenyl, indanyl, 1,2-dihydronaptha alkyl radical is one to eight carbon atoms (C1-C8), or one to lene, 1,2,3,4-tetrahydronaphthyl, and the like. Aryl groups six carbonatoms (C1-C6). Examples of alkyl groups include, are optionally substituted independently with one or more but are not limited to, methyl (Me, —CH), ethyl (Et, substituents described herein. —CHCH), 1-propyl (n-Pr, n-propyl. —CH2CHCH). The terms "heterocycle”, “heterocyclyl and "heterocyclic 2-propyl (i-Pr. i-propyl, —CH(CH)), 1-butyl (n-Bu, n-bu ring are used interchangeably herein and refer to a saturated tyl, —CHCHCHCH), 2-methyl-1-propyl (i-Bu, i-butyl, or a partially unsaturated (i.e., having one or more double —CH-CH(CH)), 2-butyl (s-Bu, s-butyl, —CH(CH) 10 and/or triple bonds within the ring) carbocyclic radical of 3 to CHCH), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH)), 20 ring atoms in which at least one ring atom is a heteroatom 1-pentyl (n-pentyl, —CH2CH2CH2CHCH), 2-pentyl selected from nitrogen, oxygen, phosphorus and Sulphur, the (—CH(CH)CHCHCH), 3-pentyl ( CH(CH2CH)), remaining ring atoms being C, where one or more ring atoms 2-methyl-2-butyl ( C(CH) CHCH), 3-methyl-2-butyl is optionally substituted independently with one or more sub (-CH(CH)CH(CH)), 3-methyl-1-butyl ( CHCHCH 15 stituents described below. A heterocycle may be a monocycle (CH)), 2-methyl-1-butyl ( CHCH(CH)CHCH), having 3 to 7 ring members (2 to 6 carbon atoms and 1 to 4 1-hexyl ( CHCH2CH2CHCH), 2-hexyl ( CH(CH) heteroatoms selected from N, O, P, and S) or a bicycle having CHCHCHCH), 3-hexyl (-CH(CHCH.) 7 to 10 ring members (4 to 9 carbon atoms and 1 to 6 heteroa (CH2CHCH)), 2-methyl-2-pentyl (—C(CH) toms selected from N, O, P, and S), for example: a bicycle4. CHCHCH), 3-methyl-2-pentyl ( CH(CH)CH(CH) 5, 5.5, 5.6, or 6.6 system. Heterocycles are described in CHCH), 4-methyl-2-pentyl ( CH(CH)CHCH(CH)), Paquette, Leo A.; “Principles of Modern Heterocyclic Chem 3-methyl-2-pentyl ( CH(CH)(CHCH)), 2-methyl-3- istry” (W. A. Benjamin, New York, 1968), particularly Chap pentyl ( CH(CHCH)CH(CH)), 2,3-dimethyl-2-butyl ters 1, 3, 4, 6, 7, and 9; “The Chemistry of Heterocyclic (—C(CH)-CH(CH)), 3.3-dimethyl-2-butyl ( CH(CH) Compounds. A series of Monographs” (John Wiley & Sons, C(CH), 1-heptyl, 1-octyl, and the like. 25 New York, 1950 to present), in particular Volumes 13, 14, 16, The term “alkenyl refers to linear or branched-chain 19, and 28; and J. Am. Chem. Soc. (1960) 82:5566. “Hetero monovalent hydrocarbon radical of two to eight carbonatoms cyclyl also includes radicals where heterocycle radicals are (C2-C8) with at least one site of unsaturation, i.e., a carbon fused with a saturated, partially unsaturated ring, or aromatic carbon, sp2 double bond, wherein the alkenyl radical may be carbocyclic or heterocyclic ring. Examples of heterocyclic optionally substituted independently with one or more sub 30 rings include, but are not limited to, pyrrolidinyl, tetrahydro stituents described herein, and includes radicals having "cis' furanyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyra and “trans” orientations, or alternatively, “E” and “Z” orien nyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, mor tations. Examples include, but are not limited to, ethylenyl or pholino, thiomorpholino, thioxanyl, piperazinyl, vinyl ( CH=CH-), allyl ( CH-CH=CH-), and the like. homopiperazinyl, azetidinyl, oxetanyl, thietanyl, homopip The term “alkynyl refers to a linear or branched monova 35 eridinyl, oxepanyl, thiepanyl, oxazepinyl, diazepinyl, thiaz lent hydrocarbon radical of two to eight carbon atoms (C2 epinyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl. C8) with at least one site of unsaturation, i.e., a carbon 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl. carbon, sp triple bond, wherein the alkynyl radical may be dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, optionally substituted independently with one or more sub pyrazolidinylimidazolinyl, imidazolidinyl, 3-azabicyclo stituents described herein. Examples include, but are not lim 40 3.1.0 hexanyl, 3-azabicyclo4.1.0 heptanyl, azabicyclo ited to, ethynyl ( C=CH), propynyl (propargyl. 2.2.2]hexanyl, 3H-indolyl quinolizinyl and N-pyridyl ureas. —CHC=CH), and the like. Spiro moieties are also included within the scope of this The term “halogen' (or halo) preferably represents chloro definition. Examples of a heterocyclic group wherein 2 ring or fluoro, but may also be bromo or iodo. carbon atoms are substituted with oxo (=O) moieties are The terms “carbocycle”, “carbocyclyl', 'carbocyclic ring 45 pyrimidinonyl and 1,1-dioxo-thiomorpholinyl. The hetero and “cycloalkyl refer to a monovalent non-aromatic, Satu cycle groups herein are optionally Substituted independently rated or partially unsaturated ring having 3 to 12 carbonatoms with one or more substituents described herein. (C3-C12) as a monocyclic ring or 7 to 12 carbon atoms as a The term "heteroaryl' refers to a monovalent aromatic bicyclic ring. Bicyclic carbocyclated having 7 to 12 atoms can radical of 5-, 6-, or 7-membered rings, and includes fused ring be arranged, for example, as a bicyclo4.5, 5.5, 5.6 or 50 systems (at least one of which is aromatic) of 5-20 atoms, 6.6 system, and bicyclic carbocycles having 9 or 10 ring containing one or more heteroatoms independently selected atoms can be arranged as a bicycle 5.6 or 6.6 system, or as from nitrogen, oxygen, and Sulphur. Examples of heteroaryl bridged systems such as bicyclo[2.2.1]heptane, bicycle groups are pyridinyl (including, for example, 2-hydroxypy 2.2.2]octane and bicycle3.2.2]nonane. Examples of mono ridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl (includ cyclic carbocycles include, but are not limited to, cyclopro 55 ing, for example, 4-hydroxypyrimidinyl), pyrazolyl, triaz pyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, olyl pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cy thiazolyl, oxadiazolyl, oxazolyl, isothiazolyl pyrrolyl, clohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl, cyclo quinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, indolyl, hexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl benzimidazolyl, benzofuranyl, cinnolinyl, indazolyl, indoliz cycloundecyl, cyclododecyl, and the like. 60 inyl, phthalazinyl, pyridazinyl, triazinyl, isoindolyl, pteridi Aryl means a monovalent aromatic hydrocarbon radical of nyl, purinyl, oxadiazolyl, triazolyl, thiadiazolyl, furazanyl. 6-20 carbon atoms (C6-C20) derived by the removal of one benzofurazanyl, benzothiophenyl, benzothiazolyl, benzoox hydrogen atom from a single carbon atom of a parent aro azolyl, quinazolinyl, quinoxalinyl, naphthyridinyl, and furo matic ring system. Some aryl groups are represented in the pyridinyl. Heteroaryl groups are optionally Substituted inde exemplary structures as 'Ar. Aryl includes bicyclic radicals 65 pendently with one or more substituents described herein. comprising an aromatic ring fused to a Saturated, partially The heterocycle or heteroaryl groups may be carbon (car unsaturated ring, or aromatic carbocyclic ring. Typical aryl bon-linked), or nitrogen (nitrogen-linked) bonded where US 8,921,361 B2 11 12 Such is possible. By way of example and not limitation, car cancer. To the extent the drug may prevent growth and/or kill bon bonded heterocycles or heteroaryls are bonded at posi existing cancer cells, it may be cytostatic and/or cytotoxic. tion 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or 6 of a For cancer therapy, efficacy can be measured, for example, by pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2, 3, assessing the time to disease progression (TTP) and/or deter 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan, tetrahy mining the response rate (RR). drofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole, The terms “cancer and "cancerous” refer to or describe the position 2, 4, or 5 of an oxazole, imidazole or thiazole, posi physiological condition in mammals that is typically charac tion 3, 4, or 5 of an isoxazole, pyrazole, or isothiazole, posi terized by unregulated cell growth. A “tumor comprises one tion 2 or 3 of an aziridine, position 2, 3, or 4 of an azetidine, or more cancerous cells. Examples of cancer include, but are position 2, 3, 4, 5, 6, 7, or 8 of a quinoline or position 1, 3, 4, 10 not limited to, carcinoma, lymphoma, blastoma, sarcoma, 5, 6, 7, or 8 of an isoquinoline. and leukaemia or lymphoid malignancies. More particular By way of example and not limitation, nitrogen bonded examples of such cancers include squamous cell cancer (e.g., heterocycles or heteroaryls are bonded at position 1 of an epithelial squamous cell cancer), lung cancer including aziridine, azetidine, pyrrole, pyrrolidine, 2-pyrroline, 3-pyr Small-cell lung cancer, non-Small cell lung cancer roline, imidazole, imidazolidine, 2-imidazoline, 3-imidazo 15 (NSCLC), adenocarcinoma of the lung and squamous car line, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline, piperi cinoma of the lung, cancer of the peritoneum, hepatocellular dine, piperazine, indole, indoline, 1H-indazole, position 2 of cancer, gastric or stomach cancer including gastrointestinal a isoindole, or isoindoline, position 4 of a morpholine, and cancer, pancreatic cancer, glioblastoma, cervical cancer, ova position 9 of a carbazole, or 3-carboline. rian cancer, liver cancer, bladder cancer, hepatome, breast The term “monocyclic heteroaryl' refers to a five- or six cancer, colon cancer, rectal cancer, colorectal cancer, membered, unsubstituted or substituted, monocyclic het endometrial or uterine carcinoma, salivary gland carcinoma, eroaryl radical which contains 1, 2, 3 or 4 ring heteroatoms kidney or renal cancer, prostate cancer, Vulval cancer, thyroid independently selected from N, O and S. The monocyclic cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, heteroaryl may be attached to the C-2 position of the pyrimi as well as head and neck cancer. dine ring according to formula Ia-Ii at any carbon (carbon 25 A “chemotherapeutic agent' is a chemical compound use linked), or nitrogen (nitrogen-linked) atom of the monocyclic ful in the treatment of cancer. Examples of known chemo heteroaryl R3 group. Monocyclic heteroaryl radicals include, therapeutic agents include trastuzumab, pertuZumab, erlo but are not limited to: 2-pyridyl, 3-pyridyl, 4-pyridyl, 3-isox tinib (TARCEVAR), Genetech/OSI Pharm.), bortezomib azolyl, 4-isoxazolyl, 5-isoxazolyl, 2-imidazolyl, 4-imida (VELCADE(R), Millennium Pharm.), fulvestrant (FASLO Zoly1,3-pyrazolyl, 4-pyrazolyl 2-pyrrolyl, 3-pyrrolyl, 2-thia 30 DEXR, AstraZeneca), Sunitib (SUTENTR), Pfizer/Sugen), Zolyl, 4-thiazolyl, 5-thiazolyl, 3-pyridazinyl, 4-pyridazinyl, letrozole (FEMARAR), Novartis), imatinib mesylate 5-pyridazinyl, 2-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, (GLEEVECR), Novartis), finasunate (VATALANIB(R), 2-pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 2-furanyl. Novartis), oxaliplatin (ELOXATINR, Sanofi), 5-FU (5-fluo 3-furanyl, 2-thienyl, 3-thienyl, 3-triazolyl, 1-triazolyl, 5-tet rouracil), leucovorin, Rapamycin (Sirolimus, RAPAM razolyl, 1-tetrazolyl, and 2-tetrazolyl. Monocyclic heteroaryl 35 UNER, Wyeth), Lapatinib (TYKERB(R, GSK572016, Glaxo are optionally substituted independently with one or more Smith Kline), Lonafarnib (SCH 66336), Sorafenib (NEXA substituents described herein. VAR, Bayer Labs), and gefitinib (IRESSAR), AstraZeneca), The term “treat' and “treatment” refer to both therapeutic AG1478, alkylating agents such as thiotepa and treatment and prophylactic or preventative measures, wherein CYTOXANR) cyclosphosphamide; alkyl sulfonates such as the object is to prevent or slow down (lessen) an undesired 40 buSulfan, improsulfan and piposulfan; aziridines Such as ben pathological change or disorder, such as the development or Zodopa, carboquone, meturedopa, and uredopa; ethylen spread of cancer. For purpose of this invention, beneficial or imines and methylamelamines including altretamine, trieth desired clinical results include, but are not limited to, allevia ylenemelamine, triethylenephosphoramide, tion of symptoms, diminishment of extent of disease, stabi triethylenethiophosphoramide and trimethylomelamine; lized (i.e., not worsening) state of disease, delay or slowing of 45 acetogenins; a camptothecin (including the synthetic analog disease progression, amelioration or palliation of the disease topotecan); bryostatin: cally statin: CC-1065 (including its state, and remission (whether partial or total), whether detect adoZelesin, carZelesin and bizelesin synthetic analogs); cryp able or undetectable. “Treatment can also mean prolonging tophycins; dolastatin; duocarmycin (including the synthetic Survival as compared to expected Survival if not receiving analogs, KW-2189 and CB1-TM1); eleutherobin: pancrat treatment. Those in need of treatment include those already 50 istatin; a sarcodictyin: Spongistatin: nitrogen mustards Such with the condition or disorder as well as those prone to have as chlorambucil, chlornaphazine, chlorophosphamide, estra the condition or disorder or those in which the condition or mustine, ifosfamide, mechlorethamine, mechlorethamine disorder is to be prevented. oxide hydrochloride, melphalan, novembichin, phenesterine, The phrase “therapeutically effective amount’ means an prednimustine, trofosfamide, uracil mustard; nitroSureas amount of a compound of the present invention that (i) treats 55 Such as carmustine, chlorozotocin, fotemustine, lomustine, or prevents the particular disease, condition, or disorder, (ii) nimustine, and ranimnustine; antibiotics such as the enediyne attenuates, ameliorates, or eliminates one or more symptoms antibiotics (e.g., calicheamicin, especially calicheamicin of the particular disease, condition, or disorder, or (iii) pre gammall and calicheamicin omegall; dynemicin, including vents or delays the onset of one or more symptoms of the dynemicin A; biphosphonates, such as clodronate; an espe particular disease, condition, or disorder described herein. In 60 ramicin; as well as neocarzinostatin chromophore and related the case of cancer, the therapeutically effective amount of the chromoprotein enediyne antibiotic chromophores), aclacino drug may be reduce the number of cancer cells; reduce the mysins, actinomycin, authramycin, aZaserine, bleomycins, tumor size: inhibit (i.e., slow to some extent and preferably cactinomycin, carabicin, carminomycin, carzinophillin, stop) cancer cell infiltration into peripheral organs; inhibit chromomycinis, dactinomycin, daunorubicin, detorubicin, (i.e., slow to Some extent and preferably stop) tumor metasta 65 6-diazol-5-oxo-L-norleucine, ADRIAMYCINR) (doxorubi sis; inhibit, to Some extent, tumor growth; and/or relieve to cin), morpholino-doxorubicin, cyanomorpholino-doxorubi Some extent one or more of the symptoms associated with the cin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epiru US 8,921,361 B2 13 14 bicin, esorubicin, idarubicin, marcellomycin, mitomycins The term “prodrug” as used in this application refers to a Such as mitomycin C, mycophenolic acid, nogalamycin, precursor or derivative form of a compound of the invention olivomycins, peplomycin, porfiromycin, puromycin, quela that may be less cytotoxic to cells compared to the parent mycin, rodorubicin, Streptonigrin, Streptozocin, tubercidin, compound or drug and is capable of being enzymatically or ubenimex, Zinostatin, Zorubicin; anti-metabolites Such as hydrolytically activated or converted into the more active methotrexate and 5-fluorouracil (5-FU); folic acid analogs parent form. The prodrugs of this invention include, but are Such as denopterin, methotrexate, pteropterin, trimetrexate; not limited to, phosphate-containing prodrugs, thiophos purine analogs such as fludarabine, 6-mercaptopurine, thia phate-containing prodrugs, Sulfate-containing prodrugs, pep miprine, thioguanine; pyrimidine analogs such as ancitabine, tide-containing prodrugs, D-amino acid-modified prodrugs, 10 glycosylated prodrugs, B-lactam-containing prodrugs, aZacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuri optionally substituted phenoxyacetamide-containing pro dine, doxifluridine, enocitabine, floXuridine; androgens Such drugs, optionally Substituted phenylacetamide-containing as calusterone, dromostanolone propionate, epitiostanol, prodrugs, 5-fluorocytosine and other 5-fluorouridine pro mepitioStane, testolactone; anti-adrenals such as aminoglute drugs which can be converted into the more active cytotoxic thimide, mitotane, triloStane; folic acid replenisher such as 15 free drug. Examples of cytotoxic drugs that can be derivatized frolinic acid; aceglatone; aldophosphamide glycoside; ami into a prodrug form for use in this invention include, but are nolevulinic acid; eniluracil; amsacrine; bestrabucil; bisant not limited to, compounds of the invention and chemothera rene; ediatraxate; defofamine; demecolcine; diaziquone; peutic agents such as described above. elfornithine; elliptinium acetate; an epothilone; etoglucid: A “metabolite' is a product produced through metabolism gallium nitrate; hydroxyurea; lentinan; lonidainine; may in the body of a specified compound or salt thereof. Metabo tansinoids such as maytansine and ansamitocins; mitogua lites of a compound may be identified using routine tech Zone; mitoxantrone; mopidanmol; nitraerine; pentostatin: niques known in the art and their activities determined using phenamet; pirarubicin; losoxantrone; podophyllinic acid; tests Such as those described herein. Such products may result 2-ethylhydrazide; procarbazine; PSKR) polysaccharide com for example from the oxidation, reduction, hydrolysis, ami plex; razoxane; rhizoxin; sizofiran, spirogermanium; tenu 25 dation, deamidation, esterification, deesterification, enzy aZonic acid; triaziquone; trichothecenes; urethane; vindesine; matic cleavage, and the like, of the administered compound. dacarbazine; mannomustine; mitobronitol; mitolactol; pipo Accordingly, the invention includes metabolites of com broman; gacytosine; arabinoside; taxoids, e.g., TAXOL(R) pounds of the invention, including compounds produced by a (paclitaxel; Bristol-Myers Squibb), ABRAXANETM (Cremo process comprising contacting a compound of this invention phor-free), albumin-engineered nanoparticle formulations of 30 with a mammal for a period of time sufficient to yield a paclitaxel, and TAXOTERE(R) (docetaxel, doxetaxel; Sanofi metabolic product thereof. Aventis); chloranmbucil: GEMZAR(R) (gemcitabine); A "liposome is a small vesicle composed of various types 6-thioguanine; mercaptopurine; methotrexate; platinum ana of lipids, phospholipids and/or surfactant, which is useful for logs such as cisplatin and carboplatin; vinblastine; etoposide; delivery of a drug (such as the PI3K and mTOR kinase inhibi ifosfamide; mitoxantrone; Vincristine; NAVELEBINE(R) (vi 35 tors disclosed herein and, optionally, a chemotherapeutic norelbine); novantrone; teniposide; ediatrexate, daunomycin; agent) to a mammal. The components of the liposome are aminopterin; capecitabine (XELODAR); ibandronate: commonly arranged inabilayerformation, similar to the lipid CP-11: topoisomerase inhibitor RFS 2000; difluoromethylor arrangement of biological membranes. nithine (DMFO); retinoids such as retinoic acid; and pharma The term “package insert” is used to refer to instructions ceutically acceptable salts; acids and derivatives of any of the 40 customarily included in commercial packages of therapeutic above. products, that contain information about the indications, Also included in the definition of “chemotherapeutic usage, dosage, administration, contraindications and/or agent” are: (i) anti-hormonal agents that act to regulate or warnings concerning the use of Such therapeutic products. inhibit hormone action on tumors such as anti-estrogens and The term "chiral' refers to molecules, which have the selective receptor modulators (SERMs), including, for 45 property of non-Superimposability of the mirror image part example, tamoxifen (including NOLVADEXOR); tamoxifen ner, while the term “achiral” refers to molecules, which are citrate), raloxifene, droloxifene, and FARESTONR) (toremi Superimposable on their mirror image partner. fine citrate); (ii) aromatase inhibitors that inhibit the enzyme The term “stereoisomers' refers to compounds, which aromatase, which regulates estrogen production in the adre have identical chemical constitution, but differ with regard to nal glands, such as, for example, 4(5)-imidazoles, 50 the arrangement of the atoms or groups in space. MEGASE(R) (megestrol acetate); AROMASINR) (exemes “Diastereomer' refers to a stereoisomer with two or more tane; Pfizer), formestanie, fadrazole, RIVISORR) (vorozole), centers of chirality and whose molecules are not mirror FEMARAR) (letrozole; Novartis), and ARIMIDEX(R) (anas images of one another. Diastereomers have different physical trozole; AstraZeneca); (iii) anti-androgens such as flutamide, properties, e.g. melting points, boiling points, spectral prop nilutamide; (iv) protein kinase inhibitors; (V) lipid kinase 55 erties, and reactivities. Mixtures of diasteremers may sepa inhibitors; (vi) antisense oligonucleotides, particularly those rate under high resolution analytical procedures such as elec which inhibit expression of genes in signaling pathways trophoresis and chromatography. implicated in aberrant cell proliferation, such as, for example, "Enantiomers' refer to two stereoisomers of a compound PKC-alpha, Rafl and H-Ras; (vii) ribozymes such as VEGF which are non-Superimposable mirror images of one another. expression inhibitors (e.g., ANGIOZYME(R) and HER2 60 Stereochemical definitions and conventions used herein expression inhibitors; (viii) vaccines such as gene therapy generally follow S. P. Parker, Ed., McRaw-Hill Dictionary of vaccines, for example, ALLOVECTINR, LEUVECTINR, Chemical Terms (1984) McGraw-Hill Book Company, New and VAXID(R); PROLEUKINR rIL-2; a topoisomerase 1 York; and Eliel, E. and Wilen, S., “Stereochemistry of inhibitor such as LURTOTECANER); ABARELIX(R) rmRH: Organic Compounds”, John Wiley & Sons, Inc., New York, (ix) anti-angiogenic agents such as bevacizumab (AVAS 65 1994. The compounds of the invention may contain asym TINR, Genentech); and (x) pharmaceutically acceptable metric or chiral centers, and therefore exist in different stere salts, acids and derivatives of any of the above. oisomeric forms. It is intended that all stereoisomeric forms US 8,921,361 B2 15 16 of the compounds of the invention, including but not limited amines, and cyclic amines, such as piperidine, morpholine to, diastereomers, enantiomers and atropisomers, as well as and piperazine, and inorganic salts derived from Sodium, mixtures thereof Such as racemic mixtures, form part of the calcium, potassium, magnesium, manganese, iron, copper, present invention. Many organic compounds existin optically Zinc, aluminium and lithium. active forms, i.e., they have the ability to rotate the plane of 5 The phrase “pharmaceutically acceptable' indicates that plane-polarized light. In describing an optically active com the Substance or composition must be compatible chemically pound, the prefixes D and L, or Rand S. are used to denote the and/or toxicologically, with the other ingredients comprising absolute configuration of the molecule about its chiral a formulation, and/or the mammal being treated therewith. center(s). The prefixes d and 1 or (+) and (-) are employed to A “solvate” refers to an association or complex of one or designate the sign of rotation of plane-polarized light by the 10 compound, with (-) or 1 meaning that the compound is more solvent molecules and a compound of the invention. levorotatory. A compound prefixed with (+) or d is dextroro Examples of solvents that form solvates include, but are not tatory. For a given chemical structure, these Stereoisomers are limited to, water, isopropanol, ethanol, methanol, DMSO, identical except that they are mirror images of one another. A ethyl acetate, acetic acid, and ethanolamine. The term specific stereoisomer may also be referred to as an enanti 15 “hydrate' refers to the complex where the solvent molecule is omer, and a mixture of Such isomers is often called an enan Water. tiomeric mixture. A 50:50 mixture of enantiomers is referred The term “protecting group' refers to a substituent that is to as a racemic mixture or a racemate. commonly employed to block or protect a particular function The term “tautomer' or “tautomeric form' refers to struc ality while reacting other functional groups on the compound. tural isomers of different energies, which are interconvertible For example, an "amino-protecting group' is a Substituent via a low energy barrier. For example, proton tautomers attached to an amino group that blocks or protects the amino include interconversions via migration of a proton, such as functionality in the compound. Suitable amino-protecting keto-enol and imin-enamine isomerizations. groups include acetyl, trifluoroacetyl, t-butoxycarbonyl The phrase “pharmaceutically acceptable salt as used (BOC), benzyloxycarbonyl and 9-fluorenylmethylenoxycar herein, refers to pharmaceutically acceptable organic or inor 25 bonyl (Fmoc). For a general description of protecting groups ganic salts of a compound of the invention. Exemplary salts and their use, see T. W. Greene, Protective Groups I Organic include, but are not limited to, Sulfate, citrate, acetate, oxalate, Synthesis, John Wiley & Sons, New York, 1991. chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid The terms “compound of this invention' and “compounds phosphate, isonicotinate, lactate, salicylate, acid citrate, tar of the present invention' and “compounds of formula (I) or trate, oleate, tannate, pantothenate, bitartrate, ascorbate, suc 30 one of formulas (Ia) to (Ii) include compounds of formula (I) cinate, maleate, gentisinate, fumarate, gluconate, glucur or one of formulas (Ia) to (Ii) and stereoisomers, geometric onate, saccharate, formate, benzoate, glutamate, isomers, tautomers, Solvates, metabolites, N-oxide deriva methanesulfonate “mesylate”, ethanesulfonate, benzene tives and pharmaceutically acceptable salts and prodrugs Sulfonate, p-toluenesulfonate, and pamoate salts. A pharma thereof. ceutically acceptable salt may involve the inclusion of 35 The term “mammal’ includes, but is not limited to, another molecule Such as an acetate ion, a Succinate ion or humans, mice, rats, guinea, pigs, monkeys, dogs, cats, horses, other counter ion. The counter ion may be any organic or cows, pigs, and sheep. inorganic moiety that stabilizes the charge on the parent com Triazine, Pyrimidine and Pyridine Analogs and their Use as pound. Furthermore, a pharmaceutically acceptable salt may Therapeutic Agents and Diagnostic Probes have more than one charged atom in its structure. Instances 40 The present invention provides triazine, pyrimidine and where multiple charged atoms are part of the pharmaceuti pyridine compounds, and pharmaceutical formulations cally acceptable salt can have multiple counter ions. Hence, a thereof, which are useful as therapeutic agents and novel pharmaceutically acceptable salt can have one or more diagnostic probes. Moreover, these compounds are poten charged atoms and/or one or more counterion. tially useful in the treatment of diseases, conditions and/or If the compound of the invention is a base, the desired 45 disorders modulated by protein kinases and lipid kinases. pharmaceutically acceptable salt may be prepared by any More specifically, the present invention provides com suitable method available in the art, for example, treatment of pounds of formula I, the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, Sulfuric acid, nitric acid, methane Sulfonic acid, phosphoric acid and the like, or with an organic 50 (I) acid, such as acetic acid, trifluoroacetic acid, maleic acid, X R3, Succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, a pyra R3. DC nosidyl acid, Such as glucuronic acid or galacturonic acid, an RS, N R3 alpha hydroxyl acid, such as citric acid or tartaric acid, an 55 amino acid, such as aspartic acid or glutamic acid, an aro 1. matic acid, such as benzoic acid or cinnamic acid, a Sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like. If the compound of the invention is an acid, the desired 60 pharmaceutically acceptable salt may be prepared by any and stereoisomers, geometric isomers, tautomers, Solvates, suitable method, for example, treatment of the free acid with metabolites, N-oxide derivatives and pharmaceutically an inorganic or organic base. Such as an amine, an alkali metal acceptable salts thereof, wherein: hydroxide or alkaline earth metal hydroxide, or the like. Illus Q=N, G=N, U—N (triazine-based compounds of formulas trative examples of suitable salts include, but are not limited 65 (Ia) to (Id)); to, organic salts derived from amino acids, such as glycine Q-C, G=C, U—N (pyridine-based compounds of formula and arginine, ammonia, primary, secondary, and tertiary (Ie));

US 8,921,361 B2 19 20 —OR, (20) —SR, (21) —SOR, (22) —SOR2, and -continued (23) —SONRR, (24) linker moiety (hydrophobic link O O O ers, hydrophilic linkers, photo-cleavable linkers, redox reac tion-cleavable linkers), (25) linker moiety with covalently bonded TAG-molecules (a TAG could be a fluorophor, biotin, COCOOO different polymer beads and different reactive groups); wherein R, and R2, are independently selected from the group consisting of (a) hydrogen, (b) Substituted or unsubsti tuted alkyl, (c) substituted or unsubstituted aryl, (d) substi tuted or unsubstituted heteroaryl, (e) substituted or unsubsti 10 tuted heterocyclyl, and (f) substituted or unsubstituted cycloalkyl, wherein R. R. and R are independently H, C-C, alkyl, C-C alkenyl, C-C alkynyl, C-C carbocyclyl, C-Coheterocyclyl, Co-Co aryl, or C-Coheteroaryl, 15 or Ro, R together with the nitrogen to which they are attached optionally form a C-C heterocyclic ring option ally containing one or more additional ring atoms selected from N, O or S, wherein said heterocyclic ring is optionally Substituted with one or more groups independently selected from oxo, CF, F, Cl, Br, I, C-C alkyl, C-C alkenyl, C-Cs alkynyl, Cs-C12 carbocyclyl, C2-Co heterocyclyl. Co-Co aryland C-Coheteroaryl; wherein Ra and Rs are independently selected from H. 25 C-C alkyl, or —(CH2)-aryl, or Ra and Rs together with the atoms to which they are attached form a saturated or partially unsaturated C-C2 carbocyclic ring wherein 30 Y is O, S, or NR; m is 0, 1, 2, 3, 4, 5 or 6: n is 1, 2, 3, 4, 5, or 6; and 35 t is 2, 3, 4, 5 or 6. Preferably, X is O (the ring containing X being morpholine) and/or

R.33 R.3's R. R. are independently selected from the 40 Structures:

45

50

55

60

where the wavy line indicates the attachment to the 4-po 65 sition of the core ring (triazine or pyrimidine orpyridinering). In another aspect of the invention, R is substituted or unsubstituted heterocyclyl, or substituted or unsubstituted US 8,921,361 B2 21 22 —O-heterocyclyl. In another aspect, R is substituted or -continued unsubstituted morpholinyl; more particularly, R is unsubsti tuted N-linked morpholinyl. In another aspect, R comprises substituted or unsubsti tuted heterocyclylalkyl, or substituted or unsubstituted het eroarylalkyl. In another aspect, R is substituted or unsubstituted tet rahydropyran or substituted or unsubstituted tetrahydropyra nyloxy. More particularly, R is unsubstituted 4-tetrahydro 10 pyranyloxy. In another aspect, R comprises Substituted or unsubsti tuted tetrahydropyran. In a more particular aspect, tetrahy dropyran comprises 4-tetrahydropyranyloxy. 15 In another aspect, R is substituted or unsubstituted tet rahydrofuran or substituted or unsubstituted tetrahydrofura nyloxy. More particularly, R is unsubstituted 3-tetrahydro furanyloxy. 2N 2N -N In another aspect, R is phenyl, wherein phenyl is unsub NH NH N y stituted or substituted with one or more groups selected from NS/ NSN N N-methylcarboxamide, isopropylsulfonylamino, methylsul fonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypropana mide, 2-methoxyacetamide, (propan-2-ol)sulfonyl, 2-amino R e 2N 2-methylpropanamide, 2-aminoacetamide, 25 N o O 2-hydroxyacetamide, methylsulfonylamino. 2-(dimethy M s S. lamino)acetamide, amino, acetylamino, carboxamide, (4-methylsulfonylpiperazino)-1-methyl, (4-methylpiper aZino)-1-methyl, hydroxymethyl, and methoxy. 30 2N e A In certain embodiments, R is pyridyl, thiazolyl, isox NH NH azolyl, oxadiazolyl, or pyrimidyl, which heterocycyl group is N unsubstituted or substituted with one or more groups selected from N-methylcarboxamide, isopropylsulfonylamino, meth ylsulfonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypro 35 panamide, 2-meth oxyacetamide, (propan-2-ol)sulfonyl, / All A, N 2-amino-2-methylpropanamide, 2-aminoacetamide, 2-hy X/ N )/ N) droxyacetamide, methylsulfonylamino. 2-(dimethylamino) acetamide, amino, acetylamino, carboxamide, (4-methylsul fonylpiperazino)-1-methyl, (4-methylpiperazino)-1-methyl, 40 where the wavy line indicates the attachment to the 4-po hydroxymethyl, and methoxy. sition of the core ring (triazine or pyrimidine orpyridinering), In another aspect of the invention, R is Substituted or and unsubstituted heterocyclyl, or substituted or unsubstituted where the monocyclic heteroaryl group is optionally Sub —O-heterocyclyl. stituted with one or more groups selected from F, Cl, Br, I, In another aspect, R comprises Substituted or unsubsti 45 —NRoR. --OR —C(O)Rio —NRC(O)R. —N(C tuted heterocyclylalkyl, or substituted or unsubstituted het (O)R). - NRC(O)NRR ls —C(=O)CR, —C(=O) eroarylalkyl. NRoR, and C-C alkyl. In another aspect R is a monocyclic heteroaryl group In certain embodiments, R is selected from the structures: selected from pyridyl, isoxazolyl, imidazolyl pyrazolyl pyr rolyl, thiazolyl pyridazinyl, pyrimidinyl, pyrazinyl, 50 oxazolyl, furanyl, thienyl, triazolyl, tetrazolyl, where the monocyclic heteroaryl group is optionally substituted with one or more groups selected from F, Cl, Br, I, —CN, —NRoR. --OR —C(O)Rio —NRoC(O)R. —N(C 55 (O)R), NRC(O)NRR, —C(=O)CR —C(=O) NRR, C1-C12 alkyl and (C1-C12 alkyl)-OR: N In a particular embodiment, R is selected from the struc n n n 60 C.2 C.N vi.)2

65 NS/ rS21 `r2 US 8,921,361 B2 23 24 -continued -continued O N N N=\ in-( HN-N N 21 5 NH NH W 2

where the wavy line indicates the attachment to the 4-po sition of the core ring (triazine or pyrimidine orpyridinering), 10 CF. and where the monocyclic heteroaryl group is optionally Sub stituted with one or more groups selected from F, Cl, Br, I, HN \ HN —NRoR. --OR —C(O)Rio —NRoC(O)R. —N(C (O)R), NRC(O)NRR, —C(=O)CR —C(=O) 15 NRoR, and C-C alkyl. In certain embodiments, R is a monocyclic or bicyclic heteroraryl group selected from the following structures: eer /S

N H

vo's-OratNSOMe /5SOMe /5 e N NH

/ US 8,921,361 B2 25 26 continued -continued N=\ 2 N 1. N H : 5 / FC eN V 10 N H NH 21 NN

O

15

2N. eer \ NH NH

HN-N US 8,921,361 B2 27 28 -continued -continued

10

21 NH2 21 NH2 15

NN N

CN NH2 / ClN NH 2

N N

/or2N NH2 / 2 NH2 25 OH

/r- As a 30 N2 Nsu-N Nulls NH2

rs r 35 21 OH 2 OH --- es 40

/ N OH 45 2. 2 /rCric OH Arts 50

55 A - "^'s-li, No 60 ar, NN - l. 65 US 8,921,361 B2 29 -continued

—C(=Y)NROR —C(=O)NRS(O)R —C(=O) NR(CRRs)..NRR, —NO. —NHR, —NRC (—Y)R. —NRC(=Y)OR, NRC(=Y)NRR, —NRS(O)Rio —NRSONRoR, -S(O)Ro, —S(O)NRR, —SC(=Y)R —SC(=Y)OR, C-C, where the wavy line indicates the site of attachment, alkyl, C-C alkenyl, C-C alkynyl, C-C carbocyclyl, In certain embodiments, the monocyclic or bicyclic het 10 C-Co heterocyclyl, Co-Co aryl, and C-Co heteroaryl; or eroaryl group is Substituted with one or more groups selected where the C-C carbocyclyl, C-Co heterocyclyl, C-Co from F. - NH, -NHCH, N(CH), —OH, - OCH, aryl, or C-Co heteroaryl is Substituted at vicinal carbon —C(O)CH, -NHC(O)CH, -N(C(O)CH), -NHC(O) atoms of the morpholine and forms a fused bicyclic morpholi NH, -COH, -CHO, —CHOH, -C(=O)NHCH, nyl: —C(=O)NH, and —CH. 15 One aspect of the invention provides compounds having where said alkyl, alkenyl, alkynyl, carbocyclyl, heterocy general formula (Ia): clyl, aryland heteroaryl are optionally substituted with one or more groups independently selected from F, Cl, Br, I, —CN,

(Ia) NRoR ls —CRRs)..NRoR, —(CRRs),C(=Y) NRR ls —(CRRs)(—C(=Y)OR, (CRRs), C R3, NRSOR, —CRRs), ORio —(CRRs), Rio —(CRRs). SOR —NRoR —NRC(=Y)Ro, R3, 1. R3 25

R l Nals R2 30 and stereoisomers, geometric isomers, tautomers, solvates, metabolites, N-oxide derivatives and pharmaceutically NRoR optionally substituted C-C alkyl, optionally sub acceptable salts thereof, wherein stituted C-C alkenyl, optionally Substituted C-C alkynyl, optionally substituted C-C carbocyclyl, optionally substi X, R. R. R. R. R. and R2 are as defined above for 35 formula (I). tuted C-Coheterocyclyl, optionally Substituted Co-Co aryl, The preferences, particular aspects and embodiments set and optionally Substituted C-Coheteroaryl; forth above for X, R. R. R. R. R. and R2 in formula (I) wherein Roc R and R2 are independently H. C-C2 apply to these substituents in formula (Ia). alkyl, C-C alkenyl, C-C alkynyl, C-C carbocyclyl, Another aspect of the invention provides a compound hav 40 C-Coheterocyclyl, Co-Co aryl, or C-Cohetero-aryl, ing general formula (Ib): or Ro, R together with the nitrogen to which they are attached optionally form a C-C heterocyclic ring option ally containing one or more additional ring atoms selected (Ib) from N, O or S, wherein said heterocyclic ring is optionally 45 Substituted with one or more groups independently selected from oxo, (CH), OR (CH2)NRR, CF, F, Cl, Br, I, r SOR, C(=O)Ro NRC(=Y)R. C(=Y)NRR, C-C alkyl, C-C alkenyl, C-C alkynyl, C-C carbocy R3, 1. R3 clyl, C-Coheterocyclyl, Co-Co aryland C-Coheteroaryl; R N1 SN 50 wherein Ra and Rs are independently selected from H. R4. l als C-C alkyl, or—(CH)-aryl, N N R2 or Ra and Rs together with the atoms to which they are O attached form a saturated or partially unsaturated C-C2 55 carbocyclic ring wherein Y is O, S, or NR; and stereoisomers, geometric isomers, tautomers, Solvates, m is 0, 1, 2, 3, 4, 5 or 6: metabolites, N-oxid derivatives and pharmaceutically accept 60 n is 1, 2, 3, 4, 5, or 6 and able salts thereof, wherein R. R. R. R. and R2 are as defined above for formula t is 2, 3, 4, 5 or 6. (I) The preferences, particular aspects and embodiments set R. R. R. and R are independently from each other forth above for Rs. R. R. R. R. and R2 in formula (I) and selected from the group consisting of 65 apply to these substituents in formula (Ib). F. Cl, Br, I, —C(C-C alkyl)-NRoR, -(CRRs). Another aspect of the invention provides a compound hav NRoR —C(RRs)NRC(=Y)Rio —(CRRs), ing general formula (IC): US 8,921,361 B2 31 32 In another aspect, R is phenyl, wherein phenyl is unsub (Ic) stituted or substituted with one or more groups selected from N-methylcarboxamide, isopropylsulfonylamino, methylsul fonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypropana C R3, mide, 2-methoxyacetamide, (propan-2-ol)sulfonyl, 2-amino 2-methylpropanamide, 2-aminoacetamide, R3, 1. R 2-hydroxyacetamide, methylsulfonylamino. 2-(dimethy lamino)acetamide, amino, acetylamino, carboxamide, (4-methylsulfonylpiperazino)-1-methyl, (4-methylpiper R l als 10 aZino)-1-methyl, hydroxymethyl, and methoxy. Y. N R In certain embodiments, R is pyridyl, thiazolyl, isoX azolyl, oxadiazolyl, or pyrimidyl, which heterocycyl group is and stereoisomers, geometric isomers, tautomers, Solvates, unsubstituted or substituted with one or more groups selected metabolites, N-oxide derivatives and pharmaceutically from N-methylcarboxamide, isopropylsulfonylamino, meth acceptable salts thereof, wherein 15 ylsulfonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypro R. R. R. R. and R2 are as defined above for formula panamide, 2-methoxyacetamide, (propan-2-ol)sulfonyl, (I) 2-amino-2-methylpropanamide, 2-aminoacetamide, 2-hy Y is O, S, NH, or a direct bond, droxyacetamide, methylsulfonylamino. 2-(dimethylamino) R" is selected from the group consisting of acetamide, amino, acetylamino, carboxamide, (4-methylsul (1) H. F. Cl, Br, I, (2) cyano, (3) nitro, (4) halogen, (5) fonylpiperazino)-1-methyl, (4-methylpiperazino)-1-methyl, substituted and unsubstituted alkyl, (6) substituted and hydroxymethyl, and methoxy. unsubstituted alkenyl, (7) substituted and unsubstituted alky In another embodiment, R' is selected from the group nyl, (8) substituted and unsubstituted aryl, (9) substituted and consisting of: (1) Substituted or unsubstituted morpholinyl, unsubstituted heteroaryl, (10) substituted and unsubstituted heterocyclyl, (11) substituted and unsubstituted cycloalkyl, (2) substituted or unsubstituted tetrahydropyranyl, and (3) (12) —C(C-C alkyl)-NRoR —(CRRS),NRoR, 25 substituted or unsubstituted tetrahydrofuranyl. —C(RRs)..NRC(=Y)Rio —(CRRs)NRS(O) In a more particular embodiment thereof. R' is N-linked Rio —CH(OR)Ro, —(CRRs), ORio, —(CRRs),S morpholinyl. In another more particular embodiment, R is (O)Rio —(CRRs), S(O)NRoR, -C(=Y)Ro, 4-tetrahydropyranyl. In another embodiment, R' is 3-tet —C(=Y)OR —C(=Y)NRR, —C(=Y)NROR, rahydrofuranyl. —C(=O)NRS(O)Ro, —C(=O)NR2 (CRRs), 30 Another aspect of the invention provides a compound hav NRR, NO. —NHR, NRC(=Y)R. —NRC ing general formula (Id): (—Y)OR, NRC(=Y)NRR, NRS(O).R. —NRSONRoR, -S(O)Rio - S(O)NRoR - SC (=Y)Ro. —SC(=Y)OR, C-C alkyl, C-C alkenyl, (Id) C-Cs alkynyl, C-C carbocyclyl, C-Co heterocyclyl, 35 Cs-Co aryl or C-Coheteroaryl, (13) linker moiety (hydro phobic linkers, hydrophilic linkers, photo-cleavable linkers, redox reaction-cleavable linkers), (14) linker moiety with covalently bonded TAG-molecules (a TAG could be a fluo rophor, biotin, different polymer beads and different reactive 40 groups) wherein Rio R, R2, R1, Ris, Y, m, n and t are as defined above for formula (I). The preferences, particular aspects and embodiments set forth above for Rs. R. R. R. and R2 in formula (I) apply to these substituents in formula (Ic). 45 In another aspect of the invention R is substituted or unsubstituted heterocyclyl, or substituted or unsubstituted —O-heterocyclyl. In another aspect, R' is substituted or and stereoisomers, geometric isomers, tautomers, Solvates, unsubstituted morpholinyl: more particularly, R is unsubsti metabolites, N-oxide derivatives and pharmaceutically tuted N-linked morpholinyl: more particularly, X is a direct 50 acceptable salts thereof, wherein link. In another more particular embodiment, R is 4-tetrahy Rs. Rs. Rs. Rs and R are as defined above for formula dropyranyl: more particularly, X is O. In another embodi (I), ment, R is 3-tetrahydrofuranyl: more particularly, X is O. W is CR, or N, wherein Rw is selected from the group In another aspect thereof, R' comprises substituted or consisting of: unsubstituted heterocyclylalkyl, or substituted or unsubsti 55 (1) hydrogen, (2) cyano, (3) halogen, (4) methyl, (5) trifluo tuted heteroarylalkyl. romethyl, (6) sulfonamido: In another aspect, R' is substituted or unsubstituted tet Rs is selected from the group consisting of: rahydropyran or substituted or unsubstituted tetrahydropyra (1) hydrogen, and (2) halogen, nyloxy. More particularly, R is unsubstituted 4-tetrahydro R is selected from the group consisting of: pyranyloxy. 60 (1) hydrogen, (2) cyano, (3) nitro, (4) halogen, (5) Substituted In another aspect thereof, R' comprises substituted or and unsubstituted alkyl, (6) substituted and unsubstituted alk unsubstituted tetrahydropyran. In a more particular aspect, enyl, (7) substituted and unsubstituted alkynyl, (8) substituted tetrahydropyran comprises 4-tetrahydropyranyloxy. and unsubstituted aryl, (9) substituted and unsubstituted het In another aspect, R' is substituted or unsubstituted tet eroaryl, (10) substituted and unsubstituted heterocyclyl, (11) rahydrofuran or substituted or unsubstituted tetrahydrofura 65 substituted and unsubstituted cycloalkyl, (12) —COR (13) nyloxy. More particularly, R is unsubstituted 3-tetrahydro —NR.R., (14) —NRCOR (15) —NRSO.R., (16) furanyloxy. —OR (17) —SR (18) —SOR (19) —SOR, and US 8,921,361 B2 33 34 (20)—SONRR, wherein R, and R, are independently (=Y)Ro. —SC(=Y)OR, C-C alkyl, C-C alkenyl, selected from the group consisting of: (a) hydrogen, (b) Sub C-C alkynyl, C-C carbocyclyl, C-C heterocyclyl, stituted or unsubstituted alkyl, (c) substituted and unsubsti Co-Co aryl or C-Coheteroaryl, (13) linker moiety (hydro tuted aryl, (d) substituted and unsubstituted heteroaryl, (e) phobic linkers, hydrophilic linkers, photo-cleavable linkers, substituted and unsubstituted heterocyclyl, and (f) substituted 5 redox reaction-cleavable linkers), (14) linker moiety with and unsubstituted cycloalky, and covalently bonded TAG-molecules (a TAG could be a fluo R is selected from the group consisting of: rophor, biotin, different polymer beads and different reactive (1) hydrogen, (2) substituted and unsubstituted alkyl, and (3) groups) wherein Rio R. R. R. Ris, Y, m, n and t are as substituted and unsubstituted cycloalkyl, (4) methyl, (5) defined above for formula (I). ethyl, (6) trifluoromethyl, (7) sulfonamid, (8) acetate, (9) 10 The preferences, particular aspects and embodiments set linker moiety (hydrophobic linkers, hydrophilic linkers, forth above for X, R. R. R. R. and R2 in formula (I) photo-cleavable linkers, redox reaction-cleavable linkers), apply to these substituents in formula (Ie). (10) linker moiety with covalently bonded TAG-molecules (a In another aspect of the invention R is substituted or TAG could be a fluorophor, biotin, different polymer beads 15 unsubstituted heterocyclyl, or substituted or unsubstituted and different reactive groups). —O-heterocyclyl. In another aspect, R' is substituted or The preferences, particular aspects and embodiments set unsubstituted morpholinyl; more particularly, R is unsubsti forth above for R. R. R. R. and R in formula (I) apply tuted N-linked morpholinyl: more particular still, X is a direct to these substituents in formula (Id). link. In another more particular embodiment, R is 4-tetrahy In a more particular embodiment, W is CH. dropyranyl: more particularly, X is O. In another embodi In another embodiment, W is N. In a more particular ment, R is 3-tetrahydrofuranyl: more particularly, X is O. embodiment thereof, R is —O. In another aspect thereof, R' comprises substituted or In another embodiment, R is selected from the group unsubstituted heterocyclylalkyl, or substituted or unsubsti consisting of: tuted heteroarylalkyl. (1) cyano, (2) nitro, (3) halogen, (4) hydroxyl, (5) amino, 25 In another aspect, R' is substituted or unsubstituted tet and (6) trifluoromethyl. In another embodiment, R is trifluo rahydropyran or substituted or unsubstituted tetrahydropyra romethyl. In another embodiment, R is cyano. nyloxy. More particularly, R is unsubstituted 4-tetrahydro Another aspect of the invention provides a compound hav pyranyloxy. ing general formula (Ie): In another aspect thereof, R' comprises substituted or 30 unsubstituted tetrahydropyran. In a more particular aspect, (Ie) tetrahydropyran comprises 4-tetrahydropyranyloxy. In another aspect, R' is substituted or unsubstituted tet rahydrofuran or substituted or unsubstituted tetrahydrofura nyloxy. More particularly, R is unsubstituted 3-tetrahydro C 35 furanyloxy. R3, 1. R3 In another aspect, R is phenyl, wherein phenyl is unsub stituted or substituted with one or more groups selected from N-methylcarboxamide, isopropylsulfonylamino, methylsul R - als 40 fonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypropana Y. U R2 mide, 2-methoxyacetamide, (propan-2-ol)sulfonyl, 2-amino 2-methylpropanamide, 2-aminoacetamide, and stereoisomers, geometric isomers, tautomers, Solvates, 2-hydroxyacetamide, methylsulfonylamino. 2-(dimethy metabolites, N-oxide derivatives and pharmaceutically lamino)acetamide, amino, acetylamino, carboxamide, acceptable salts thereof, wherein 45 (4-methylsulfonylpiperazino)-1-methyl, (4-methylpiper aZino)-1-methyl, hydroxymethyl, and methoxy. In certain embodiments, R is pyridyl, thiazolyl, isox azolyl, oxadiazolyl, or pyrimidyl, which heterocycyl group is X, R. R. R. R. and R are as defined above for unsubstituted or substituted with one or more groups selected formula (I), 50 from N-methylcarboxamide, isopropylsulfonylamino, meth Y is O, S, NH, or a direct bond, ylsulfonyl, 2-hydroxy-2-methylpropanamide, 2-hydroxypro R" is selected from the group consisting of panamide, 2-methoxyacetamide, (propan-2-ol)sulfonyl, (1) H. F. Cl, Br, I, (2) cyano, (3) nitro, (4) halogen, (5) 2-amino-2-methylpropanamide, 2-aminoacetamide, 2-hy substituted and unsubstituted alkyl, (6) substituted and droxyacetamide, methylsulfonylamino. 2-(dimethylamino) unsubstituted alkenyl, (7) substituted and unsubstituted alky 55 acetamide, amino, acetylamino, carboxamide, (4-methylsul nyl, (8) substituted and unsubstituted aryl, (9) substituted and fonylpiperazino)-1-methyl, (4-methylpiperazino)-1-methyl, unsubstituted heteroaryl, (10) substituted and unsubstituted hydroxymethyl, and methoxy. heterocyclyl, (11) substituted and unsubstituted cycloalkyl, In another embodiment, R' is selected from the group (12) —C(C-C alkyl)-NRoR —(CRRS),NRoR, consisting of: (1) Substituted or unsubstituted morpholinyl, —C(RRs)..NRC(=Y)Rio —(CRRs)NRS(O) 60 (2) substituted or unsubstituted tetrahydropyranyl, and (3) Rio —CH(OR)R —(CRRs), OR —(CRRs),S substituted or unsubstituted tetrahydrofuranyl. (O)Rio —(CRRs), S(O)NRoR, -C(=Y)Ro, In a more particular embodiment thereof, R' is N-linked —C(=Y)OR, —C(=Y)NRR, —C(=Y)NROR, morpholinyl. In another more particular embodiment, R is —C(=O)NRS(O)Ro, —C(=O)NR2 (CRRs), 4-tetrahydropyranyl. In another embodiment, R' is 3-tet NRR, NO. —NHR, NRC(=Y)R. —NRC 65 rahydrofuranyl. (—Y)OR, NRC(=Y)NRR, NRS(O).R. Another aspect of the invention provides compounds hav —NRSONRoR, -S(O)Rio - S(O)NRoR - SC ing one of general formulas (If) and (Ig): US 8,921,361 B2 35 36 and unsubstituted aryl, (9) substituted and unsubstituted het (If) eroaryl, (10) substituted and unsubstituted heterocyclyl, (11) substituted and unsubstituted cycloalkyl, (12) —COR (13) —COR (14) —CONR.R. (15) —NR.R., (16) C —NRCOR (17) —NRSO.R. (18) —OCOR (19) RS, N R3 —OR (20) —SR, (21) —SOR, (22) —SOR2, and (23) —SONRR, (24) linker moiety (hydrophobic link Y, No. ers, hydrophilic linkers, photo-cleavable linkers, redox reac tion-cleavable linkers), (25) linker moiety with covalently R-( es2 10 bonded TAG-molecules (a TAG could be a fluorophor, biotin, Z N R different polymer beads and different reactive groups); I wherein R, and R, are independently selected from the group consisting of (a) hydrogen, (b) Substituted or unsubsti r (Ig) tuted alkyl, (c) substituted or unsubstituted aryl, (d) substi 15 tuted or unsubstituted heteroaryl, (e) substituted or unsubsti RS, N R3 tuted heterocyclyl, and (f) substituted or unsubstituted cycloalkyl, Z n G or Ro, R together with the nitrogen to which they are attached optionally form a C-C heterocyclic ring option R-{Y. 4.2. R ally containing one or more additional ring atoms selected from N, O or S, wherein said heterocyclic ring is optionally Substituted with one or more groups independently selected and stereoisomers, geometric isomers, tautomers, Solvates, from oxo, CF, F, Cl, Br, I, C-C alkyl, C-C alkenyl, C-Cs metabolites, N-oxide derivatives and pharmaceutically alkynyl, Cs-C12 carbocyclyl, C2-Co heterocyclyl. Co-Co acceptable salts thereof, wherein 25 aryland C-Coheteroaryl; G is N or G is C, wherein Ra and Rs are independently selected from H. X, R. R. R. R. and R2 are as defined above for C-C alkyl, or —(CH2)-aryl, formula (I), or Ra and Rs together with the atoms to which they are Y, is O, S or SO, attached form a saturated or partially unsaturated C-C2 Z is N or CR, wherein R, is selected from the group 30 carbocyclic ring consisting of: wherein (1) linker moiety (hydrophobic linkers, hydrophilic linkers, Y is O, S, or NR; photo-cleavable linkers, redox reaction-cleavable linkers), m is 0, 1, 2, 3, 4, 5 or 6: (2) linker moiety with covalently bonded TAG-molecules (a n is 1, 2, 3, 4, 5, or 6; and TAG could be a fluorophor, biotin, different polymer beads 35 t is 2, 3, 4, 5 or 6. and different reactive groups), The preferences, particular aspects and embodiments set Rs is selected from the group consisting of: forth above for X. R. R. R. R. and R2 in formula (I) (1) H. F. Cl, Br, I, (2) cyano, (3) nitro, (4) halogen, (5) apply to these substituents in formulas (If) and (Ig). substituted and unsubstituted alkyl, (6) substituted and In particular aspect of the invention, Rs is Substituted or unsubstituted alkenyl, (7) substituted and unsubstituted alky 40 unsubstituted heterocyclyl, or substituted or unsubstituted nyl, (8) substituted and unsubstituted aryl, (9) substituted and —O-heterocyclyl. In another aspect, Rs is substituted or unsubstituted heteroaryl, (10) substituted and unsubstituted unsubstituted morpholinyl; more particularly, R is unsubsti heterocyclyl, (11) substituted and unsubstituted cycloalkyl, tuted N-linked morpholinyl. (12) —C(C-C alkyl)-NRoR —(CRRS),NRoR, In another aspect, Rs comprises Substituted or unsubsti —C(RRs)..NRC(=Y)Rio —(CRRs)NRS(O) 45 tuted heterocyclylalkyl, or substituted or unsubstituted het Ro. —CH(OR)Rio —(CRRs), ORio —(CRRs),S eroarylalkyl. (O).R. —(CRRs), S(O)NRR, —C(=Y)Ro, In another aspect, Rs is substituted or unsubstituted tet —C(=Y)OR —C(=Y)NRR, —C(=Y)NROR, rahydropyran or substituted or unsubstituted tetrahydropyra —C(=O)NRS(O).R. —C(=O)NR(CRRs), nyloxy. More particularly, R is unsubstituted 4-tetrahydro NRR, NO. —NHR, NRC(=Y)R. —NRC 50 pyranyloxy. (—Y)OR, NRC(=Y)NRR, NRS(O).R. In another aspect thereof Rs comprises Substituted or —NRSONRoR, -S(O)Rio - S(O)NRoR - SC unsubstituted tetrahydropyran. In a more particular aspect, (=Y)Ro. —SC(=Y)OR, C-C alkyl, C-C alkenyl, tetrahydropyran comprises 4-tetrahydropyranyloxy. C-C alkynyl, C-C carbocyclyl, C-C heterocyclyl, In another aspect, Rs is substituted or unsubstituted tet Co-Co aryl or C-Coheteroaryl, (13) linker moiety (hydro 55 rahydrofuran or substituted or unsubstituted tetrahydrofura phobic linkers, hydrophilic linkers, photo-cleavable linkers, nyloxy. More particularly, R is unsubstituted 3-tetrahydro redox reaction-cleavable linkers), (14) linker moiety with furanyloxy. covalently bonded TAG-molecules (a TAG could be a fluo In another aspect, Rs is optionally Substituted phenyl, rophor, biotin, different polymer beads and different reactive wherein phenyl is substituted with one or more groups groups), wherein, 60 selected from N-methylcarboxamide, isopropylsulfony R and R are independently H, C-C alkyl, C-Cs lamino, methylsulfonyl, 2-hydroxy-2-methylpropanamide, alkenyl, C-C alkynyl, C-C carbocyclyl, C-Co hetero 2-hydroxypropanamide, 2-methoxyacetamide, (propan-2-ol) cyclyl. Co-Co aryl, or C-Coheteroaryl, Sulfonyl, 2-amino-2-methylpropanamide, 2-aminoaceta Ro is selected from the group consisting of: mide, 2-hydroxyacetamide, methylsulfonylamino. 2-(dim (1) hydrogen, (2) cyano, (3) nitro, (4) halogen, (5) Substituted 65 ethylamino)acetamide, amino, acetylamino, carboxamide, and unsubstituted alkyl, (6) substituted and unsubstituted alk (4-methylsulfonylpiperazino)-1-methyl, (4-methylpiper enyl, (7) substituted and unsubstituted alkynyl, (8) substituted aZino)-1-methyl, hydroxymethyl, and methoxy. US 8,921,361 B2 37 38 In certain embodiments, Rs is optionally substituted C-Cs alkynyl, C-C carbocyclyl, C-Co heterocyclyl, pyridyl, optionally substituted thiazolyl, optionally substi C-C aryl or C-C heteroaryl, (13) linker moiety (hydro tuted isoxazolyl, optionally Substituted oxadiazolyl, or phobic linkers, hydrophilic linkers, photo-cleavable linkers, optionally substituted pyrimidyl. redox reaction-cleavable linkers), (14) linker moiety with In another embodiment, Rs is selected from the group covalently bonded TAG-molecules (a TAG could be a fluo consisting of: (1) Substituted or unsubstituted morpholinyl, rophor, biotin, different polymer beads and different reactive (2) substituted or unsubstituted tetrahydropyranyl, and (3) groups), wherein Rio R. R. R. Rs. Y. m., n and t are as substituted or unsubstituted tetrahydrofuranyl. defined above for formula (I). In a more particular embodiment thereof. Rs is N-linked In another aspect of the invention, R is substituted or morpholinyl. In another more particular embodiment, Rs is 10 unsubstituted heterocyclyl, or substituted or unsubstituted 4-tetrahydropyranyl; in another embodiment, Rs is 3-tetrahy —O-heterocyclyl. In another aspect, Ro is substituted or drofuranyl. unsubstituted morpholinyl; more particularly, R is unsubsti For the use of Rio, see general reaction scheme 45. tuted N-linked morpholinyl. Another aspect of the invention provides compounds hav In another aspect, R, comprises substituted or unsubsti ing general formulas (Ih) and (Ili): 15 tuted heterocyclylalkyl, or substituted or unsubstituted het eroarylalkyl. In another aspect, Ro is substituted or unsubstituted tet rahydropyran or substituted or unsubstituted tetrahydropyra nyloxy. More particularly, R is unsubstituted 4-tetrahydro (Ih) pyranyloxy. In another aspect thereof, R., comprises substituted or unsubstituted tetrahydropyran. In a more particular aspect, tetrahydropyran comprises 4-tetrahydropyranyloxy. In another aspect, Ro is substituted or unsubstituted tet 25 rahydrofuran or substituted or unsubstituted tetrahydrofura nyloxy. More particularly, R is unsubstituted 3-tetrahydro furanyloxy. In another aspect, R is optionally substituted phenyl, wherein phenyl is substituted with one or more groups 30 selected from N-methylcarboxamide, isopropylsulfony lamino, methylsulfonyl, 2-hydroxy-2-methylpropanamide, (Ii) 2-hydroxypropanamide, 2-methoxyacetamide, (propan-2-ol) Sulfonyl, 2-amino-2-methylpropanamide, 2-aminoaceta mide, 2-hydroxyacetamide, methylsulfonylamino. 2-(dim 35 ethylamino)acetamide, amino, acetylamino, carboxamide, (4-methylsulfonylpiperazino)-1-methyl, (4-methylpiper aZino)-1-methyl, hydroxymethyl, and methoxy. In certain embodiments, Ro is optionally substituted pyridyl, optionally substituted thiazolyl, optionally substi 40 tuted isoxazolyl, optionally substituted oxadiazolyl, or optionally substituted pyrimidyl. In another embodiment, R is selected from the group and stereoisomers, geometric isomers, tautomers, Solvates, consisting of: (1) Substituted or unsubstituted morpholinyl, metabolites, N-oxide derivatives and pharmaceutically (2) substituted or unsubstituted tetrahydropyranyl, and (3) acceptable salts thereof, wherein 45 substituted or unsubstituted tetrahydrofuranyl. G is N or G is C, In a more particular embodiment thereof, R is N-linked X, R. R. R. R. and R are as defined above for morpholinyl. In another more particular embodiment, R is formula (I), 4-tetrahydropyranyl; in another embodiment, Ro is 3-tetrahy Y is O, S, SO, NH, or a direct bond, drofuranyl. L is C, N or N-oxide, 50 The compounds of the invention may contain asymmetric Ro is selected from the group consisting of: or chiral centers, and therefore exist in different stereoiso (1) H. F. Cl, Br, I, (2) cyano, (3) nitro, (4) halogen, (5) meric forms. It is intended that all stereoisomeric forms of the substituted and unsubstituted alkyl, (6) substituted and compounds of the invention, including but not limited to, unsubstituted alkenyl, (7) substituted and unsubstituted alky diastereomers, enantiomers and atropisomers, as well as mix nyl, (8) substituted and unsubstituted aryl, (9) substituted and 55 tures thereof Such as racemic mixtures, form part of the unsubstituted heteroaryl, (10) substituted and unsubstituted present invention. heterocyclyl, (11) substituted and unsubstituted cycloalkyl, In addition, the present invention embraces all geometric (12) —C(C-C alkyl)-NRoR —(CRRS),NRoR, and positional isomers. For example, if a compound of the —C(RRs)..NRC(=Y)Rio —(CRRs)NRS(O) invention incorporates a double bond or a fused ring, the cis Ro. —CH(OR)Rio —(CRRs), ORio —(CRRs),S 60 and trans-forms, as well as mixtures thereof, are embraced (O).R. —(CRRs), S(O)NRR, —C(=Y)Ro, within the scope of the invention. Both the single positional —C(=Y)OR —C(=Y)NRR, —C(=Y)NROR, isomers and mixture of positional isomers are also within the —C(=O)NRS(O).R. —C(=O)NR(CRRs), Scope of the present invention. NRR, NO. —NHR, NRC(=Y)R. —NRC In the structures shown herein, where the stereochemistry of (—Y)OR, NRC(=Y)NRR, NRS(O).R. 65 any particular chiral atom is not specified, then all stereoiso —NRSONRoR, -S(O)Rio - S(O)NRoR - SC mers are contemplated and included as the compounds of the (=Y)Ro. —SC(=Y)OR, C-C alkyl, C-C alkenyl, invention. Where stereochemistry is specified by a solid US 8,921,361 B2 39 40 wedge or dashed line representing a particular configuration, according to a further aspect of the invention there is provided then that stereoisomer is so specified and defined. a compound library comprising at least 2 compounds, or The compounds of the present invention may exist in pharmaceutically acceptable salts thereof. unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it For illustrative purposes, Schemes 1-52 show general is intended that the invention embraces both solvated and methods for preparing the compounds of the present inven unsolvated forms. tion as well as key intermediates. For a more detailed descrip The compounds of the invention may also existin different tion of the individual reaction steps, see the Examples here inbelow. Those skilled in the art will appreciate that other tautomeric forms (tautomers), and all Such forms are 10 synthetic routes may be used to synthesize the compounds of embraced with the scope of the invention. the invention. Although specific starting materials and The present invention also embrace a isotopically-labelled reagents are depicted in the Schemes and discussed below, compounds of the present invention which are identical to other starting materials and reagents can be easily Substituted those recited herein, but for the fact that one or more atoms are to provide a variety of derivatives and/or reaction conditions. replaced by an atom having an atomic mass or mass number 15 In addition, many of the compounds prepared by the methods different from the atomic mass or mass number usually found described below can be further modified in light of this dis in nature. All isotopes of any particular atom or element as closure using conventional chemistry well known to those specified are contemplated within the scope of the com skilled in the art. pounds of the invention, and their uses. Exemplary isotopes that can be incorporated into compounds of the invention In preparing compounds of the invention, protection of include isotopes of hydrogen, carbon, nitrogen, oxygen, remote functionality (e.g., primary or secondary amine) of phosphorus, Sulphur, fluorine, chlorine and iodine, such as intermediates may be necessary. The need for Such protection 2H, *H, IC, 13C, 14C, 13N, 15O, 17O, 18O, 32P 33P. 35s, 18F will vary depending on the nature of the remote functionality Cl, 'I and 'I. Certain isotopically-labelled compounds and the conditions of the preparation methods. Suitable of the present invention (e.g., those labelled with Hand ''C) 25 amino-protecting groups include acetyl, trifluoroacetyl, t-bu are useful in compound and/or substrate tissue distribution toxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9-fluo assays. Tritiated (3H) and carbon-14 (''C) isotopes are useful renylmethyleneoxycarbonyl (Fmoc). The need for such pro for their ease of preparation and detectability. Further, sub tection is readily determined by one skilled in the art. For a stitution with heavier isotopes such as deuterium (i.e., H) general description of protecting groups and their use, see T. may afford certain therapeutic advantages resulting from 30 W. Greene, Protective Groups in Organic Synthesis, John greater metabolic stability (e.g., increased in Vivo half-life or Wiley & Sons, New York, 1991. reduced dosage requirements) and hence may be preferred in some circumstances. Positron emitting isotopes such as O, 'N, ''Cand F are useful for positron emission tomography Schemes for Preparing the Compounds of the Invention (PET) studies to examine Substrate receptor occupancy. Iso 35 topically labelled compounds of the present invention can generally be prepared by following procedures analogous to Scheme 1 those disclosed in the Schemes and/or in the Examples herein Hal below, by substituting an isotopically labelled reagent for a non-isotopically labeled reagent. 40 Preparation of Compounds of the Invention N 1. N The compounds of the invention may be synthesized by synthetic routes that include processes analogous to those Hal l Nals Hal R well known in the chemical arts, particularly in light of the 1 description contained herein. The starting materials are gen 45 erally available from commercial sources such as Aldrich Chemicals or are readily prepared using methods well known Scheme 1 shows a general method for preparation of the to those skilled in the art (e.g., prepared by methods generally triazine intermediate 2 from 2,4,6-trihalo-1,3,5-triazine reagent (1), wherein Hal is Cl, Br, or I; and R is as defined for described in Louis F. Fieser and Mary Fieser, Reagents for 50 Organic Synthesis, v. 1-19, Wiley, N.Y. (1967-1999 ed.), or formula Ia-d compounds, or precursors or prodrugs thereto. Beilsteins Handbuch der organischen Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, including Supplements (also avail Scheme 2 able via the Beilstein online database). In certain embodiments, the compounds of the invention 55 Hal may be readily prepared using procedures well-known to prepare triazines and other heterocycles, which are described N 1. N in: Comprehensive Heterocyclic Chemistry, Editors Katritzky and Rees, Pergamon Press, 1984. l als Compounds of the invention may be prepared singly or as 60 Hal N Hal R compound libraries comprising at least 2, for example 5 to 1 1,000 compounds, or 10 to 100 compounds. Libraries of compounds of the invention may be prepared by a combina Scheme 2 shows a general method for preparation of the torial split and mix' approach or by multiple parallel synthe 65 triazine intermediate 3 from 2,4,6-trihalo-1,3,5-triazine ses using either solution phase or solid phase chemistry, by reagent (1), wherein Hal is Cl, Br, or I; and R is as defined for procedures well known to those skilled in the art. Thus formula Ia-d compounds, or precursors or prodrugs thereto. US 8,921,361 B2 41 42 lyst may be any that is typically used for Suzuki-type cross Scheme 3 couplings, such as PdCl(PPh), Pd(PPh), Pd(OAc), O PdCl(dppf)-DCM, Pd(dba)/Pt-Bu) (Owens et al. (2003) Bioorganic & Med. Chem. Letters 13:4143-4145; Molander et al. (2002) Organic Letters 4(11):1867-1870; U.S. Pat. No. 1. O 6,448,433).

Scheme 6 10 C ) Scheme 3 shows a general method for selectively displac 15 1. (Hy)-B(ORs) N ing a halide from bis-halo triazine intermediate 2 with mor n 6 1.n pholine in an organic solvent to prepare morpholino triazine N Pd-catalyst N intermediate 4 compounds, wherein Halis Cl, Br, or I; and R. is as defined for formula Ia-d compounds, or precursors or R N Hal R N R prodrugs thereto. 4 8

Scheme 6 shows a general method for Suzuki-type cou pling of a 1-halo morpholino triazine intermediate 4 with a Scheme 4 25 cyclicheteroarylboronate acid (Rs—H) orester (R'-alkyl) reagent 6 to prepare the cyclic heteroaryl (Hy) compounds (8) of formulas Ia, wherein Hal is Cl, Br, or I; and R and Rare O as defined for formula Ia compounds, or precursors or pro drugs thereto. es 1. 30 Scheme 7

35

ON 1. (Hy)-B(ORs) N1 NN 6 Scheme 4 shows a general method for selectively displac 40 ing two halides from tris-halo triazine 1 with morpholine in an l als HePd-catalyst organic solvent to prepare bis-morpholino triazine interme N N Hal diate 5 compounds, wherein Hal is Cl, Br, or I.

45

Scheme 5 R N 1s, (Hy)-B(ORs) 50 O N 1. -e- R - als% Hal Pd-catalyst6 3 7 55 Scheme 5 shows a general method for Suzuki-type cou pling of a 1-halo triazine intermediate 3 with a cyclic het eroarylboronate acid (Rs—H) or ester (R-alkyl) reagent 6 to prepare the cyclic heteroaryl (Hy) compounds (7) of for 60 mulas Ia, wherein Hal is Cl, Br, or I; and R and R are as Scheme 7 shows a general method for Suzuki-type cou defined for formula Ia compounds, or precursors or prodrugs pling of a 1-halobis-morpholino triazine intermediate 5 with thereto. For reviews of the Suzuki reaction, see: Miyaura et al. a cyclic heteroaryl boronate acid (Rs—H) or ester (1995) Chem. Rev.95:2457-2483; Suzuki, A. (1999).J. Orga (R'-alkyl) reagent 6 to prepare the cyclic heteroaryl (Hy) nomet. Chem. 576:147-168; Suzuki, A. in Metal-Catalyzed 65 compounds (9) of formulas Ib, wherein Hal is Cl, Br, or I; and Cross-Coupling Reactions, Diederich, F. Stang, P. J. Eds. R is as defined for formula Ib compounds, or precursors or VCH, Weinheim, Del. (1998), pp. 49-97. The palladium cata prodrugs thereto. US 8,921,361 B2 43 44 Scheme 11 shows a general method for preparation of the Scheme 8 triazine intermediate 12 from triazine intermediate 13, wherein Hal is Cl, Br, or I; and R' and Y are as defined for 1s.Hal formula Ic compounds, or precursors or prodrugs thereto.

Hal l Nals Hal R Ny. l Nals Hal Scheme 12

1 10 10 (Hy)-B(ORs) Scheme 8 shows a general method for preparation of the 6 triazine intermediate 10 from 2,4,6-trihalo-1,3,5-triazine -- reagent (1), wherein Hal is Cl, Br, or I; and R', and Y are as 15 R Ny l Nals H Pd-catalyst defined for formula Ic compounds, or precursors or prodrugs thereto.

Scheme 9

25

Scheme 9 shows a general method for preparation of the 30 Scheme 12 shows a general method for Suzuki-type cou triazine intermediate 11 from 2,4,6-trihalo-1,3,5-triazine pling of a triazine intermediate 11 with a cyclic heteroaryl reagent (1), wherein Hal is Cl, Br, or I; and R' and Y are as boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to defined for formula Ic compounds, or precursors or prodrugs prepare the cyclic heteroaryl (Hy) compounds (14), wherein thereto. Hal is Cl, Br, or I; R' and Y are as defined for formula Ic 35 compounds and R is as defined for formula I compounds, or Scheme 10 precursors or prodrugs thereto. Scheme 13

Hal O 40 s -e- 1. O (Hy)-B(ORs) R Ny. us Nals Hal 6 s 1. -- 10 R Ny.us Nals Hal 45 Pd-catalyst 12 R" Ny. us Nals Hal 12 Scheme 10 shows a general method for preparation of the O triazine intermediate 12 from triazine intermediate 10, wherein Hal is Cl, Br, or I; and R', and Y are as defined for formula Ic compounds, or precursors or prodrugs thereto. ON

Scheme 11 55 s R Ny.ul Nals R2 O O 15 60 - - - - Scheme 13 shows a general method for Suzuki-type cou pling of a triazine intermediate 12 with a cyclic heteroaryl s s boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to Hal l Nals Hal R" Ny. us Nals Hal prepare the cyclic heteroaryl (Hy) compounds (15), wherein 13 12 65 Hal is Cl, Br, or I; R' and Y are as defined for formula Ic compounds and R is as defined for formula I compounds, or precursors or prodrugs thereto. US 8,921,361 B2 45 46 Scheme 14 shows a general method for Suzuki-type cou Scheme 14 pling of a triazine intermediate 12 with a cyclic heteroaryl boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to prepare cyclic heteroaryl (Hy) compounds (17), wherein R' and Y are as defined for formula Ic compounds. Rs. R. R. and Wareas defined for formula Id compounds, or precursors C1. (Hy)-B(ORs) or prodrugs thereto. 6 N n N -- 10 R l 2es Pd-catalyst n Y. N Hal Scheme 16 12

15 CN

CN s -- R Ny. N 2 Sw s R6 isNy. use N Sw Rs Nals NH2 25 16 Rs N 1. NH2 16

30 ON Scheme 14 shows a general method for Suzuki-type cou pling of a triazine intermediate 12 with a cyclic heteroaryl boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to prepare cyclic heteroaryl (Hy) compounds (16), wherein R' s R6 R 2 and Y areas defined for formula Ic compounds, Rs. Rand W 35 Ny. N Sw are as defined for formula Id compounds, or precursors or prodrugs thereto. 2 R7 Rs N N1 17 Scheme 15 40 C Scheme 16 shows a general method for preparation of triazine compounds 17 from triazine compounds 16, wherein 1. (Hy)-B(ORs) 45 6 R" and Y are as defined for formula Ic compounds, Rs. R. N n N -es R, and W are as defined for formula Id compounds, or pre R l els2 Pd-catalyst cursors or prodrugs thereto. Ny. N Hal 50 12

Scheme 17

Hal Hal 55 ON N -e- N 2 2 s N R6 Hal N Hal R N Hal 18 19 R Ny. N 2 Sw 60 2 R7 Rs N N1 Scheme 17 shows a general method for preparation of the 17 65 pyridine intermediate 19 from 2,4,6-trihalopyridine reagent (18), wherein Hal is Cl, Br, or I; and R is as defined for formula I compounds, or precursors or prodrugs thereto. US 8,921,361 B2 47 48

Scheme 18 Scheme 22 Hal R Hal Hal 5 N e- N N Ho- N

2 2 2 2 Hal N Hal Hal N Hal R N Hal R N R 18 20 10 19 22

Scheme 18 shows a general method for preparation of the Scheme 22 shows a general method for preparation of the pyridine intermediate 20 from 2,4,6-trihalopyridine reagent pyridine intermediate 22 from dihalopyridine reagent 19, (18), wherein Hal is Cl, Br, or I; and R is as defined for 15 wherein Hal is C1, Br, or I; and R is as defined for formula I formula I compounds, or precursors or prodrugs thereto. compounds, or precursors or prodrugs thereto.

Scheme 23 2O - SchClCle. 19 R R Hal R N -b- N N -e- N 25 2 2 2 2 Hal N Hal R N Hal Hal N Hal R N Hal 20 21 18 21 30 Scheme 23 shows a general method for preparation of the Scheme 19 shows a general method for preparation of the pyridine intermediate 21 from dihalopyridine reagent 20, pyridine intermediate 21 from 2,4,6-trihalopyridine reagent wherein Hal is Cl, Br, or I; and R is as defined for formula I (18), wherein Hal is Cl, Br, or I; and R is as defined for compounds, or precursors or prodrugs thereto. formula I compounds, or precursors or prodrugs thereto. 35 Scheme 21 Scheme 20 O Hal Hal C D 40 N -- N Hal N

Hal N2 Hal R N 2 R N N 18 22 2 2 45 Hal N Hal Hal N Hal 18 23 Scheme 20 shows a general method for preparation of the pyridine intermediate 22 from 2,4,6-trihalopyridine reagent (18), wherein Hal is Cl, Br, or I; and R is as defined for Scheme 21 shows a general method for preparation of the formula I compounds, or precursors or prodrugs thereto. 50 pyridine intermediate 23 from 2,4,6-trihalopyridine (18) and morpholine reagents, wherein Hal is Cl, Br, or I; or precursors or prodrugs thereto. Scheme 21 Hal R I 55 Scheme 22

N -e- N Hal

2 2 R N Hal R N Hal 60 Hal N 19 21 2 N -- r N Hal Scheme 21 shows a general method for preparation- of the Hal N 2 Hal su pyridine intermediate 21 from dihalopyridine reagent 19, 65 18 24 wherein Hal is Cl, Br, or I; and R is as defined for formula I compounds, or precursors or prodrugs thereto. US 8,921,361 B2 49 Scheme 22 shows a general method for preparation of the Scheme 26 pyridine intermediate 24 from 2,4,6-trihalopyridine (18) and morpholine reagents, wherein Hal is Cl, Br, or I; or precursors Hal (Hy)-B(ORs) R2 or prodrugs thereto. N 6 N Pd-catalyst Scheme 23 2 2 R N R R N R O 22 28 10 O Scheme 26 shows a general method for Suzuki-type cou pling of a 1-halo pyridine intermediate 22 with a cyclic het Hal N eroarylboronate acid (Rs—H) orester (Rs alkyl) reagent 6 15 to prepare the cyclic heteroaryl (Hy) compounds (28) of 4. Hal formula I, wherein Hal is Cl, Br, or I; and R and R are as N Hs r defined for formula I compounds, or precursors or prodrugs thereto. Hal N 2 Hal su 18 25 2O Scheme 27 Scheme 23 shows a general method for preparation of the pyridine intermediate 25 from 2,4,6-trihalopyridine (18) and morpholine reagents, wherein Hal is Cl, Br, or I. 25 ON (Hy)-B(ORs) 6 Scheme 24 N -asPd-catalyst Hal 2 30 r N N Hal N Hess su 2 Hal N Hal 18 Hal 35 25 C N

r 2 r N n 40 r 2 r N N R 26 su 29 45 Scheme 24 shows a general method for preparation of the pyridine intermediate 26 from 2,4,6-trihalopyridine (18) and Scheme 27 shows a general method for Suzuki-type cou morpholine reagents, wherein Hal is Cl, Br, or I. pling of a dimorpholino pyridine intermediate 25 with a 50 cyclic heteroarylboronate acid (Rs—H) orester (Rs-alkyl) Scheme 25 reagent 6 to prepare the cyclic heteroaryl (Hy) compounds (29) of formula I, wherein Hal is Cl, Br, or I; and R is as R (Hy)-B(ORs) R defined for formula I compounds, or precursors or prodrugs thereto. N 6 N Pd-catalyst 55 Scheme 28 2 2 R N Hal R N R Hal 21 27 (Hy)-B(ORs) N 60 6 -> Scheme 25 shows a general method for Suzuki-type cou 2 Pd-catalyst pling of a 1-halo pyridine intermediate 21 with a cyclic het eroarylboronate acid (Rs—H) or ester (Rs alkyl) reagent 6 r N N N ~ to prepare the cyclic heteroaryl (Hy) compounds (27) of formula I, wherein Hal is Cl, Br, or I; and R and R are as 65 su Nu. defined for formula I compounds, or precursors or prodrugs 26 thereto. US 8,921,361 B2 51 52 -continued formula Ie compounds and R is as defined for formula I compounds, or precursors or prodrugs thereto.

Scheme 32 Hal Hal

N -e- N 30 10 R" 2 R 2 Ny. N Hal Ny. N R Scheme 28 shows a general method for Suzuki-type cou 32 35 pling of a dimorpholino pyridine intermediate 26 with a cyclic heteroarylboronate acid (Rs—H) orester (Rs-alkyl) 15 reagent 6 to prepare the cyclic heteroaryl (Hy) compounds Scheme 32 shows a general method for preparation of the (30) of formula I, wherein Hal is Cl, Br, or I; and R is as pyridine intermediate 35 from dihalopyridine reagent (32), defined for formula I compounds, or precursors or prodrugs wherein Hal is Cl, Br, or I; R' and Y are as defined for thereto. formula Ie compounds and R is as defined for formula I compounds, or precursors or prodrugs thereto.

Scheme 29 Scheme 33 O 25

Hal CN D Hal N N -- N 30 R 2 R 2 Scheme 29 shows a general method for preparation of the Ny. N Hal Ny. N Hal pyridine intermediate 32 from 2,4,6-trihalopyridine reagent 32 36 (18), wherein Hal is Cl, Br, or I; and R' and Y are as defined for formula Ie compounds, or precursors or prodrugs thereto. 3 Scheme 33 shows a general method for preparation of the morpholino pyridine intermediate 36 from dihalopyridine Scheme 30 reagent, wherein Hal is Cl, Br, or I; and R' and Y are as defined for formula (e compounds, or precursors or prodrugs Hal 40 thereto.

Scheme 34 2 Hal Hal N Hal Hal N Hal 45 18 33 N Hs R 2 Scheme 30 shows a general method for preparation of the Ny. N Hal pyridine intermediate 33 from 2,4,6-trihalopyridine reagent 50 (18), wherein Hal is Cl, Br, or I; and R' and Y are as defined 32 for formula Ie compounds or precursors or prodrugs thereto. Hal

Scheme 31 55 R r 2 Hal Ny. N N ~

-e- Nul 37 2 R"i 2 60 Hal n Hal 34 Scheme 34 shows a general method for preparation of the morpholino pyridine intermediate 37 from dihalopyridine Scheme 31 shows a general method for preparation of the 65 reagent, wherein Hal is Cl, Br, or I; and R' and Y are as pyridine intermediate 34 from dihalopyridine reagent, defined for formula Ie compounds, or precursors or prodrugs wherein Hal is Cl, Br, or I; R and Y are as defined for thereto. US 8,921,361 B2 53 54 -continued R Scheme 35 Ny. O C r O N 2 He r N N Hal

10 su r 2 R r 2 38 Hal N Hal Ny1 NN Hal 23 36 Scheme 37 shows a general method for preparation of the 15 morpholino pyridine intermediate 38 from morpholino diha Scheme 35 shows a general method for preparation of the lopyridine reagent 24, wherein Hal is Cl, Br, or I; and R' and morpholino pyridine intermediate 36 from morpholino diha Y are as defined for formula Ie compounds or precursors or lopyridine reagent 23, wherein Hal is Cl, Br, or I; and R' and prodrugs thereto. Y are as defined for formula Ie compounds, or precursors or prodrugs thereto. Scheme 38

R (Hy)-B(ORs) N 6 Scheme 36 25 Pd-catalyst Hal R" 2 n Y. N Hal N 34 R He 2 30 r N N Hal N su R 2 n Y. N R 24 35 39

Hal Scheme 38 shows a general method for Suzuki-type cou 21 pling of a pyridine intermediate 34 with a cyclic heteroaryl 40 boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to Sa R" prepare the cyclic heteroaryl (Hy) compounds 39 of formula r N N y1 I, wherein Hal is Cl, Br, or I; R', and Y are as defined for formula Ie compounds and R and R2 are as defined for su formula I compounds, or precursors or prodrugs thereto. 37 45

Scheme 39 Scheme 36 shows a general method for preparation of the Hal morpholino pyridine intermediate 37 from morpholino diha (Hy)-B(ORs) 50 lopyridine reagent 24, wherein Hal is Cl, Br, or I; and R' and N 6 Y are as defined for formula Ie compounds, or precursors or Pd-catalyst prodrugs thereto. R" 2 n Y. N R 35 55 R2 Scheme 37 Hal N R 2 N 60 n Y. N R -e- 2 N N Hal Scheme 39 shows a general method for Suzuki-type cou su 65 pling of a pyridine intermediate 35 with a cyclic heteroaryl 24 boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to prepare the cyclic heteroaryl (Hy) compounds 40 of formula US 8,921,361 B2 55 56 I, wherein Hal is Cl, Br, or I; R and Y are as defined in formula Ie compounds and R and R are as defined for formula I compounds, or precursors or prodrugs thereto. Scheme 42 R" n Y

Scheme 40 (Hy)-B(ORs) N 6 O -e- Pd-catalyst 2 C D (Hy)-B(ORs) N N Hal N 6 -e- Pd-catalyst su N 38

R 2 15 Ny. N Hall R n Y 36

r 2 r N N R2 N su 43 2 25 N R 41 Scheme 42 shows a general method for Suzuki-type cou pling of a pyridine intermediate 38 with a cyclic heteroaryl boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to Scheme 40 shows a general method for Suzuki-type cou 30 prepare the cyclic heteroaryl (Hy) compounds 43 of formula pling of a pyridine intermediate 36 with a cyclic heteroaryl Ie, wherein Hal is Cl, Br, or I: R' and Y are as defined for boronate acid (Rs—H) or ester (Rs-alkyl) reagent 6 to formula Ie compounds and R is as defined for formula I prepare the cyclic heteroaryl (Hy) compounds 41 of formula compounds, or precursors or prodrugs thereto. Ie, wherein Hal is Cl, Br, or I; R' and Y are as defined for formula Ie compounds and R is as defined for formula I 35 compounds, or precursors or prodrugs thereto. Scheme 43 Scheme 41 R8 Y, CO2R10 40 Hal ( , He (Hy)-B(ORs) N N 6 NH2 Hos 44 R 2 Pd-catalyst 45 n Y. N N ~ O

Y Nul NH -- 37 R 50 R8 - & 1s N N O H 46 Hall R r2 n Y. N N ~ 55 Null Y, NN 2 42 * {N N Hal 48 60 Scheme 41 shows a general method for Suzuki-type cou Rs N CO2R10 pling of a pyridine intermediate 37 with a cyclic heteroaryl ne f boronate acid (Rs—H) or ester (Rs alkyl) reagent 6 to Y, prepare the cyclic heteroaryl (Hy) compounds 42 of formula NH2 Ie, wherein Hal is Cl, Br, or I; R' and Y are as defined for 65 formula Ie compounds and R is as defined for formula I 45 compounds, or precursors or prodrugs thereto. US 8,921,361 B2 57 58 -continued oxazolopyrimidine intermediates 48 and 49 with morpholine O under basic conditions in an organic solvent to prepare 5-halo, 7-morpholino thiazolo- and oxazolopyrimidine compounds N NH -e- 50 and 51, wherein Y, is O or S. Hal is Cl, Br, or I; and Rs is R-( 1. 5 as defined for formula Ifg. Y; N O H 47 Hal Scheme 45 10

N n N 2 * {Y; N Hal ON 15 Y, NN RC(O)Z10C(O) K base Scheme 43 shows a general method for preparation of the H N als thiazolopyrimidine and oxazolopyrimidine intermediates 48 N N Hal and 49 from 5-carboxyester, 4-aminothiazole (X=S) and 52 oxazole (X=O), and 4-carboxyester, 5-aminothiazole (X—S) and oxazole (X—O) reagents, respectively 44 and 45, wherein Y, is O or S. Hal is Cl, Br, or I; Rs and Ro are as defined for formula Ifg. 25 ON Scheme 44 O O Y, NN Hal C D R10) {N % Hal Y, NN N 54 Rs { -e-H N 2 N N Hal 48 35 O CN O N NN RC(O)Z10C(O) H ( 2.2 base Y, NN Y; N Hal R8 { als 53 N N Hal 50 45 O

Hal C D CN N n N N 50 R-( --- O N n 2 N Y; N Hal 49 R10)-K Y; 21S O 55 55

O Scheme 45 shows a general method for derivatizing the 2-position of thiazolopyrimidine and oxazolopyrimidine N N N compounds 52 and 53 where Rs is H. Treating 52 and 53 with 60 a lithiating reagent to remove the 2-position proton, followed 2 * {Y; N Hal by adding an acylating reagent RC(O)Zwhere Z is a leaving group, such as halide, NHS ester, carboxylate, or dialky 51 lamino, gives 5-halo, 7-morpholino, 2-acyl thiazolopyrimi 65 dine and oxazolopyrimidine compounds 54 and 55, wherein Scheme 44 shows a general method for selectively displac Y, is O or S. Hal is Cl, Br, or I; and Rio is as defined for ing a 7-halide from bis-halothiazolopyrimidine and bis-halo formula If-g. US 8,921,361 B2 59 Scheme 47 Scheme 46

5 O CN N (Hy)-B(ORs) Cytochrom NN oxidation Y, NN — - -- R Pd-catalyst 8 & 10 Nals Hy N Nals Hal 54 50 nucleophilic attack to the reactive Michael system and fast 15 degradation of the molecule in cells

CN ON ON Y, n N Cytochrom V R-{N als S n oxidation \, n N Hy { K 52 N N Hy N N Hy 25 55 { no nucleophilic57 attack possible, cmpd. more stable in C cells N (Hy)-B(ORs) 30 Scheme 47 shows a general method for cellular sulfur N SN — - oxidation of thieno and thiazolo pyrimidine compounds 56 Rs —{ els Pd-catalyst and 57 through the cytochroms. After sulfur oxidation of 2 thiophene pyrimidine compounds 54 (IP of Piramed/Roche), Y; N Hal 35 the thiophene ring acts as an activated Michael system, where 51 at the 7-position of the ring a nucleophilic attack is possible and therefore a fast cellular modification of thienopyrimidine compounds 54. In comparison with thieno pyrimidines, thia Zolo pyrimidine compounds 55 are chemically more stable CN 40 through the N-atom at the 7-position and no possibility for a nucleophilic attack after sulfur oxidation.

N n N Scheme 48

2 R{Y; N Hy 45 53 ON -e- Scheme 46 shows a general method for Suzuki-type cou Y, n N pling of a 5-halopyrimidine intermediate (50 and 51) with a 50 cyclic heteroarylboronate acid (Rs—H) or ester (Rs-alkyl) & als reagent 6 to prepare the 5-cyclic heteroaryl (Hy), 7-mor N Hy 58 pholino thiazolopyrimidine and oxazolopyrimidine com pounds (52 and 53) of formulas If-g, wherein Y, is O or S. Hal is Cl, Br, or I; and Rs is as defined for formula If-g com 55 pounds, or precursors or prodrugs thereto. For reviews of the Suzuki reaction, see: Miyaura et al. (1995) Chem. Rev. 95:2457-2483; Suzuki, A. (1999).J. Organomet. Chem. 576: 147-168; Suzuki, A. in Metal-Catalyzed Cross-Coupling 60 Reactions, Diederich, F. Stang, P. J. Eds. VCH, Weinheim, Del. (1998), pp. 49-97. The palladium catalyst may be any that is typically used for Suzuki-type cross-couplings, such as PdCl(PPh), Pd(PPh), Pd(OAc), PdCl(dppf)-DCM, 59 Pd(dba)/Pt Bu) (Owens et al. (2003) Bioorganic & Med. 65 Chem. Letters 13:4143-4145; Molander et al. (2002) Organic Scheme 48 shows a general method for preparation of Letters 4(11):1867-1870; U.S. Pat. No. 6,448,433). linker-tag-modified fused pyrimidine compounds 59 from US 8,921,361 B2 61 62 morpholino intermediate 58, wherein Hy is a cyclic het eroaryl fragment, Y, and Z are as defined for compounds of Scheme 50 formula Ifg, linker and tag are as defined for formula I compounds, or precursors or prodrugs thereto.

Scheme 49

61 69

Ro SC O 45 & / NH Scheme 50 shows a general method for selectively displac L He- ing a 4-halide from bis-halo fused pyrimidine intermediates Y 1s 66 and 67 with morpholine under basic conditions in an N O organic solvent to prepare 2-halo, 4-morpholino fused pyri 50 midine compounds 68 and 69, whereinY is O or S. Hal is Cl, Br, or I; and R and L are as defined for formula Ih-i.

Scheme 51

(Hy)-B(ORs) Scheme 49 shows a general method for preparation of 6 fused pyrimidine intermediates 64 and 65 from for example -e- 2-carboxamid, 3-aminobenzofuran and 3-aminofuropyridine Pd-catalyst (X—O, L–C) and 2-carboxamid, 3-aminobenzothiophene and 3-aminothienopyridine (X=S, LC), respectively 60 and 61, wherein Y. is O or S. Hal is Cl, Br, or I; R and L are as defined for formula Ih-i. US 8,921,361 B2 63 64 -continued -continued

15 Scheme 52 shows a general method for preparation of (Hy)-B(ORs) linker-tag-modified fused pyrimidine compounds 75 from 6 morpholino intermediate 74, wherein Hy is a cyclic het -e- eroaryl fragment, Y and L are as defined for formula Ih-i Pd-catalyst compounds and linker and tag are as defined for formula I compounds, or precursors or prodrugs thereto. General Preparative Procedures General Procedure A Triazine Substitution:

C 25

He N 2 C N C R N C 30 A solution of 2,4,6-trichloro-1,3,5-triazine (1.00 g, 5.42 mmol. 1.0 eq.) in dioxane (15 ml) at room temperature was Scheme 51 shows a general method for Suzuki-type cou treated with diisopropylethylamine (1.03 ml, 5.96 mmol. 1.1 35 eq.) and dropwise with aminopyridine (1.1 eq.) and stirred for pling of a 2-halopyrimidine intermediate (70 and 71) with a 2 h. The dioxane was evaporated in vacuo, and the residue cyclic heteroarylboronate acid (Rs—H) or ester (Rs-alkyl) partitioned between HO (15 ml) and CHCl (15 ml). The reagent 6 to prepare the 2-cyclic heteroaryl (Hy), 4-mor organic layer was separated and the aqueous layer further pholino fused pyrimidine compounds (72 and 73) of formulas extracted with CHCl (2x10 ml). The combined extracts Ih-i, wherein Y is O or S. Hal is CI, Br, or I; and R and L are 40 were dried (MgSO) and concentrated in vacuo. Purification as defined for formula Ih-i compounds, or precursors or pro by column chromatography gave the title compound. drugs thereto. For reviews of the Suzuki reaction, see: General Procedure A-1 Triazine Substitution: Miyaura et al. (1995) Chem. Rev.95:2457-2483; Suzuki, A. (1999) J. Organomet. Chem. 576:147-168; Suzuki, A. in Metal-Catalyzed Cross-Coupling Reactions, Diederich, F., 45 Stang, P.J., Eds. VCH, Weinheim, Del. (1998), pp. 49-97. The C CN palladium catalyst may be any that is typically used for Suzuki-type cross-couplings, such as PdCl2(PPh3), Pd(PPh), Pd(OAc), PdCl(dppf)-DCM, Pd(dba)/Pt p 1s, Bu) (Owens et al. (2003) Bioorganic & Med. Chem. Letters 50 13:4143-4145; Molander et al. (2002) Organic Letters 4(11): R N C R N C 1867-1870; U.S. Pat. No. 6,448,433). Scheme 52 A solution of bis-chloro triazine compound (100 mg. 390 55 umol. 1.0 eq.) in dioxane (1 ml) at room temperature was treated with diisopropylethylamine (0.10 ml, 590 umol. 1.5 eq.) and morpholine (0.05 ml, 590 umol. 1.5 eq.) and stirred at room temperature for 2 h. The dioxane was evaporated in vacuo, and the residue partitioned between H2O (5 ml) and 60 CH2Cl2 (5 ml). The organic layer was separated and the aqueous layer further extracted with CHCl (2x2 ml). The combined extracts were dried (MgSO) and concentrated in vacuo. Purification by column chromatography gave the desired compound. 65 Note: for some of compounds it was necessary to add an 74 additional amount of morpholine (0.3 eq.) after 12 h reaction time and then the reaction mixture was heated 70° C. for 8 h. US 8,921,361 B2 65 66 General Procedure A-2 Triazine Substitution: -continued

C C ses s O 2 -s, ls C Nn O l N elsC 10 N 21 als NH2 247 mg of NaH (60% in mineral oil, 10.3 mmol. 0.95 eq.) was added to a solution of 2-pyridinemethanol (940 ul, 9.76 15 mmol. 0.90 eq.) in THF (20 ml) at room temperature and stirred for 30 minutes. After cooling to -78° C., 2.4.6- trichloro-1,3,5-triazine (2.00 g, 10.84 mmol. 1.0 eq.) was added dropwise and the reaction allowed to warm to room temperature and stirred for 3 h. Saturated aqueous NHCl (20 S NH2 ml) was added and the mixture extracted with EtOAc (20 ml). The organic phase was dried (MgSO) and evaporated in R-{ es2 Pd-catalyst vacuo. Column chromatography gave desired product. N N C

General Procedure B Suzuki Coupling: 25 8O OO S n N R-Q N NH2 N 4. NN N N Pd-catalyst 35 l als Nals NH2 r N Hal 81

76 40 Y. r N CN als N NH2 45 Pd-catalyst l s2 r N NN 82 50 su N els NH2 77 CN O O 55 Y. O n N o1 NN 2 N es N n N N 2 60 Pd-catalyst Nals NH2 2 83

65 The Suzuki-type coupling reaction is useful to attach a cyclic heteroaryl at the 6-position of the triazine or pyridine ring (see Schemes 7 and 27), 5- or 6-position of the pyrimi US 8,921,361 B2 67 68

dine ring (see Schemes 46 and 51). Generally, 4,4'-(6-chloro -continued 1,3,5-triazine-2,4-diyl)dimorpholine (77), 4,4'-(6-chloropy ridine-2,4-diyl)dimorpholine (79), 4-(5-chlorothiazolo 4.5- dpyrimidin-7-yl)morpholine (81) and 2-chloro-4- morpholinobenzofuro3,2-dipyrimidine (83) may be 5 combined with boronic acid pinacol ester (4.0 eq.) in 1.2- dimethoxyethane and 2M NaCO (3:1) for 15 minutes. A catalytic amount, or more, of a palladium reagent, Such as dichloro 1,1'-bis(diphenylphosphino) ferrocene palladium (II) (0.025 eq.) was added and the high pressure glass vessel 10 containing the mixture was bubbled with argon gas and sealed. A variety of boronic acids or boronic esters can be used in place of the pinacol boronic ester indicated. Also 87 alternatively, the nitrogen of the pyrimidin-2-amine may be protected, for example with a tetrahydropyranyl group. The 15 7-morpholino-5-(pyrimidin-5-yl)thiazolo 4,5-dipyrimi reaction mixture was then heated at 90° C. for 15 h or more, dine-2-carboxylic acid (84) or pyridinylfuranopyrimidine 86 cooled down and diluted with ethyl acetate. The organic solu is treated with 1.5 eq. HATU, 3 eq. of an alkylamine tion was washed with mixture of water:NaCO:NHOH (R-NH) and 3 eq. of DIPEA in DMF to approximately 0.1 (NH4OH conc. 32% in water)=5:4:1, NHCl (sat.) and brine, M concentration. The reaction is stirred until complete and dried over MgSO filtered and concentrated. The residue was extracted in ethylacetate with saturated bicarbonate solution purified by silica gel flash column chromatography or if nec one time. The organic layer is dried, filtered and concentrated essary by reverse phase HPLC. to yield the crude intermediate. This intermediate is purified via silica column chromatography or preparative TLC to General Procedure CAmide Coupling: 25 yield product 85 or 87. General Procedure C-1 Amide Coupling: O C D 30 N

HO R O S NN R-NH2 -e- HATU (1.5 eq.) ) { DIPEA (3 eq.) HO N 4. NN 35 DMF

N e 84 88

He HATU (1.5 eq.) DIPEA (3 eq.) DMF US 8,921,361 B2 69 70

-continued -continued

R 1 NR C(OMe) (10 eq.) AcOH (1 eq.) Na(OAc)3BH (1.5 eq.) Dichloroethane

O

15 O

91

4-(2-(piperazin-1-ylmethyl)-5-(pyrimidin-5-yl)thiazolo 4,5-dipyrimidin-7-yl)morpholine (88) or pyridinylfuropyri 95 midine 90 is treated with 1.5 eq. HATU, 3 eq. of carboxylic 25 acid (RCOH) and 3 eq. of DIPEA in DMF to approximately 0.1 M concentration. The reaction is stirred until complete 7-morpholino-5-(pyrimidin-5-yl)thiazolo 4,5-dipyrimi and extracted in ethyl acetate with saturated bicarbonate solu dine-2-carbaldehyde (92) or pyridinylfuropyrimidine 94 was tion one time. The organic layer is dried, filtered and concen dissolved to a 0.2M concentration in dichloroethane. To this trated to yield the crude intermediate that was used without 30 Solution was added 1.5 to 2.0 equivalents of an amine further purification or if necessary that was purified by pre (R'R''NH), 10 equivalents of trimethylorthoformate, and 1 parative TLC. equivalent of acetic acid. The mixture was allowed to stir for 2-6 hours prior to adding 1.5 equivalents of sodium triac General Procedure D Reductive Amination: etoxyborohydride. Following 12 to 16 hours of stirring the 35 reaction was poured into saturated Sodium bicarbonate and extracted several times with ethyl acetate. This intermediate was either purified on silica gel or used crude in the next reaction. O General procedure E Sulfonamide Formation: 40 O R 1 NR O (1.5-2 eq.) N 1N R" O S NN He 45 y-K C(OMe) (10 eq.) f / NN (2 eq.) 2 AcOH (1 eq.) OSS DIPEA (3 eq.) H N N SN Na(OAc);BH (15 eq.) / M 2 DCM e Dichloroethane C N nN. N 50 92 N 96 O 55 N O S NN Ost, \( 2 60 VR" 2 N 97

65 93 7-morpholino-5-(pyrimidin-5-yl)thiazolo 4,5-dipyrimi dine-2-sulfonyl chloride (96) was suspended in 1 mL of meth US 8,921,361 B2 71 72 ylene chloride before addition of 2 eq. of amine (R'R''NH) wise separable a desired product, unreacted starting material, and 3 eq. of DIPEA. The reactions were monitored by TLC reaction by product, or the like. Such reagents include adsor until complete. The crude reaction mixtures were diluted with bents or absorbents such as activated carbon, molecular ethyl acetate, extracted with Saturated ammonium chloride sieves, ion exchange media, or the like. Alternatively, the and back-extracted once with ethyl acetate. The organic lay reagents can be acids in the case of a basic material, bases in ers were combined and concentrated to dryness. The crude the case of an acidic material, binding reagents such as anti sulfonamide intermediates 97 were used directly in the sub bodies, binding proteins, selective chelators such as crown sequent Suzuki couplings ethers, liquid/liquidion extraction reagents (LIX), or the like. General Procedure F Aldehyde Synthesis: Selection of appropriate methods of separation depends on 10 the nature of the materials involved. For example, boiling point and molecular weight in distillation and Sublimation, O presence or absence of polar functional groups in chromatog raphy, stability of materials in acidic and basic media in multiphase extraction, and the like. One skilled in the art will O apply techniques most likely to achieve the desired separa tion. Diastereomeric mixtures can be separated into their indi nBuLI, THF vidual diastereomers on the basis of their physical chemical Hs differences by methods well known to those skilled in the art, cN 4. nN. Such as by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an 4. appropriate optically active compound (e.g., chiral auxiliary 98 O Such as a chiral alcohol or Mosher's acid chloride), separating 25 the diastereomers and converting (e.g., hydrolyzing) the indi vidual diastereoisomers to the corresponding pure enanti omers. Also, Some of the compounds of the present invention O may be atropisomers (e.g., Substituted biaryls) and are con sidered as part of this invention. Enantiomers can also be 30 separated by use of a chiral HPLC column. A single stereoisomer, e.g., an enantiomer, Substantially NN free of its stereoisomer may be obtained by resolution of the racemic mixture using a method Such as formation of diaste 2 reomers using optically active resolving agents (Eliel, E. and ) or N 35 Wilen, S. "Stereochemistry of Organic Compounds. John 99 Wiley & Sons, Inc., New York, 1994; Lochmuller, C. H., (1975) J. Chromatogr., 113(3):283-302). Racemic mixtures of chiral compounds of the invention can be separated and To a suspension of 4-(5-(pyrimidin-5-yl)thiazolo 4.5-d isolated by any Suitable method, including: (1) formation of pyrimidin-7-yl)morpholine (6.85 mmol. 1.0 eq.) in dry THF 40 ionic, diastereomeric salts with chiral compounds and sepa (40 mL) at -78°C. was added a 2.5 M solution of n-butyl ration by fractional crystallization or other methods, (2) for lithium (nBuLi) in hexane (1.2 eq.). After stirring for 1 h, dry mation of diastereomeric compounds with chiral derivatizing DMF (1.5 eq.) was added. The reaction mixture was stirred reagents, separation of the diastereomers, and conversion to for 1 h at -78°C. and then warmed slowly to room tempera the pure stereoisomers, and (3) separation of the Substantially ture. After a further 2 h at room temperature the reaction 45 pure or enriched Stereoisomers directly under chiral condi mixture was poured onto ice/water yielding a precipitate that tions. See: “Drug Stereochemistry, Analytical Methods and was collected by filtration and air-dried to yield the title Pharmacology. Irving W. Wainer, Ed., Marcel Dekker, Inc., compound. New York (1993). Methods of Separation Under method (1), diastereomeric salts can be formed by In the methods of preparing the compounds of this inven 50 reaction of enantiomerically pure chiral bases such as bru tion, it may be advantageous to separate reaction products cine, quinine, ephedrine, Strychnine, C.-methyl-3-phenyl from one another and/or from starting materials. The desired ethylamine (amphetamine), and the like with asymmetric products of each step or series of steps are separated and/or compounds bearing acidic functionality. Such as carboxylic purified (hereinafter separated) to the desired degree of acid and Sulfonic acid. The diastereomeric salts may be homogeneity by the techniques common in the art. Typically 55 induced to separate by fractional crystallization or ionic chro Such separations involve multiphase extraction, crystalliza matography. For separation of the optical isomers of amino tion from a solvent or solvent mixture, distillation, sublima compounds, addition of chiral carboxylic or Sulfonic acids, tion, or chromatography. Chromatography can involve any Such as camphorsulfonic acid, tartaric acid, mandelic acid, or number of methods including, for example: reverse-phase lactic acid can result information of the diastereomeric salts. and normal phase; size exclusion; ion exchange; high, 60 Alternatively, by method (2), the substrate to be resolved is medium and low pressure liquid chromatography methods reacted with one enantiomer of a chiral compound to form a and apparatus; Small scale analytical; simulated moving bed diastereomeric pair (E. and Wilen, S. "Stereochemistry of (SMB) and preparative thin or thick layer chromatography, as Organic Compounds”, John Wiley & Sons, Inc., 1994, p. well as techniques of Small scale thin layer and flash chroma 322). Diastereomeric compounds can be formed by reacting tography. 65 asymmetric compounds with enantiomerically pure chiral Another class of separation methods involves treatment of derivatizing reagents, such as menthyl derivatives, followed a mixture with a reagent selected to bind to or render other by separation of the diastereomers and hydrolysis to yield the US 8,921,361 B2 73 74 pure or enriched enantiomer. A method of determining optical sis, restenosis, psoriasis, allergic disorders, inflammation, purity involves making chiral esters, such as a menthyl ester, neurological disorders, a hormone-related disease, condi e.g., (-) menthyl chloroformate in the presence of base, or tions associated with organ transplantation, immunodefi Mosher ester, C.-methoxy-O-(trifluoromethyl)phenyl acetate ciency disorders, destructive bone disorders, proliferative (Jacob III. J. Org. Chem. (1982) 47:4165), of the racemic disorders, infectious diseases, conditions associated with cell mixture, and analyzing the "H NMR spectrum for the pres death, thrombin-induced platelet aggregation, chronic myel ence of the two atropisomeric enantiomers or diastereomers. ogenous leukemia (CML), liver disease, pathologic immune Stable diastereomers of atropisomeric compounds can be conditions involving T cell activation, and CNS disorders in a separated and isolated by normal- and reverse-phase chroma patient. In one embodiment, a human patient is treated with a tography following methods for separation of atropisomeric 10 compound of formula Ia-danda pharmaceutically acceptable naphthyl-isoquinolines (WO96/15111). By method (3), a carrier, adjuvant, or vehicle, wherein said compound of for racemic mixture of two enantiomers can be separated by mula Ia-d is present in an amount to detectably inhibit PB chromatography using a chiral stationary phase (“Chiral Liq kinase activity. uid Chromatography” (1989) W. J. Lough, Ed., Chapman and Cancers which can be treated according to the methods of Hall, New York: Okamoto, J. Chromatogr., (1990) 513:375 15 this invention include, but are not limited to, breast, ovary, 378). Enriched or purified enantiomers can be distinguished cervix, prostate, testis, genitourinary tract, esophagus, larynx, by methods used to distinguish other chiral molecules with glioblastoma, neuroblastoma, stomach, skin, keratoacan asymmetric carbonatoms, such as optical rotation and circu thoma, lung, epidermoid carcinoma, large cell carcinoma, lar dichroism. non-Small cell lung carcinoma (NSCLC), Small cell carci Administration of Compounds of the Invention noma, lung adenocarcinoma, bone, colon, adenoma, pan The compounds of the invention may be administered by creas, adenocarcinoma, thyroid, follicular carcinoma, undif any route appropriate to the condition to be treated. Suitable ferentiated carcinoma, papillary carcinoma, seminoma, routes include oral, parenteral (including Subcutaneous, intra melanoma, sarcoma, bladder carcinoma, liver carcinoma and muscular, intravenous, intraarterial, intradermal, intrathecal biliary passages, kidney carcinoma, myeloid disorders, lym and epidural), transdermal, rectal, nasal, topical (including 25 phoid disorders, hairy cells, buccal cavity and pharynx (oral), buccal and Sublingual), vaginal, intraperitoneal, intrapulmo lip, tongue, mouth, pharynx, Small intestine, colon-rectum, nary and intranasal. For local immunosuppressive treatment, large intestine, rectum, brain and central nervous system, the compounds may be administered by intralesional admin Hodgkin’s and leukemia. istration, including perfusing or otherwise contacting the Cardiovascular diseases which can be treated according to graft with the inhibitor before transplantation. It will be 30 the methods of this invention include, but are not limited to, appreciated that the preferred route may vary with for restenosis, cardiomegaly, atherosclerosis, myocardial infarc example the condition of the recipient. Where the compound tion, and congestive heart failure. is administered orally, it may be formulated as a pill, capsule, Neurodegenerative disease which can be treated according tablet, etc. with a pharmaceutically acceptable carrier or to the methods of this invention include, but are not limited to, excipient. Where the compound is administered parenterally, 35 Alzheimer's disease, Parkinson's disease, amyotrophic lat it may be formulated with a pharmaceutically acceptable eral sclerosis, Huntington's disease, and cerebral ischemia, parenteral vehicle and in a unit dosage injectable form, as and neurodegenerative disease caused by traumatic injury, detailed below. glutamate neurotoxicity and hypoxia. A dose to treat human patients may range from about 10 mg Inflammatory diseases which can be treated according to to about 1000 mg of formula Ia-d compound. A typical dose 40 the methods of this invention include, but are not limited to, may be about 100 mg to about 300 mg of the compound. A rheumatoid arthritis, psoriasis, contact dermatitis, and dose may be administered once a day (QID), twice per day delayed hypersensitivity reactions. (BID), or more frequently, depending on the pharmacokinetic Another aspect of this invention provides a compound of and pharmacodynamic properties, including absorption, dis this invention for use in the treatment of the diseases or tribution, metabolism, and excretion of the particular com 45 conditions described herein in a mammal, for example, a pound. In addition, toxicity factors may influence the dosage human, Suffering from Such disease or condition. Also pro and administration regimen. When administered orally, the vided is the use of a compound of this invention in the prepa pill, capsule, or tablet may be ingested daily or less frequently ration of a medicament for the treatment of the diseases and for a specified period of time. The regimen may be repeated conditions described herein in a warm-blooded animal. Such for a number of cycles of therapy. 50 as a mammal, for example a human, Suffering from Such Methods of Treatment with Compounds of the Invention disorder. Compounds of the present invention are useful for treating Pharmaceutical Formulations diseases, conditions and/or disorders including, but not lim In order to use a compound of this invention for the thera ited to, those characterized by over expression of lipid peutic treatment (including prophylactic treatment) of mam kinases, e.g. PI3 kinase. Accordingly, another aspect of this 55 mals including humans, it is normally formulated in accor invention includes methods of treating or preventing diseases dance with standard pharmaceutical practice as a or conditions that can be treated or prevented by inhibiting pharmaceutical composition. According to this aspect of the lipid kinases, including PI3. In one embodiment, the method invention there is provided a pharmaceutical composition comprises administering to a mammal in need thereofathera comprising a compound of this invention in association with peutically effective amount of a compound of formula Ia-d, or 60 a pharmaceutically acceptable diluent or carrier. a stereoisomer, geometric isomer, tautomer, Solvate, metabo A typical formulation is prepared by mixing a compound lite, N-oxide derivative orpharmaceutically acceptable salt or of the present invention and a carrier, diluent or excipient. prodrug thereof. Suitable carriers, diluents and excipients are well known to Diseases and conditions treatable according to the methods those skilled in the art and include materials such as carbo of this invention include, but are not limited to, cancer, stroke, 65 hydrates, waxes, water soluble and/or swellable polymers, diabetes, hepatomegaly, cardiovascular disease, Alzheimer's hydrophilic or hydrophobic materials, gelatin, oils, solvents, disease, cystic fibrosis, autoimmune diseases, atherosclero water and the like. The particular carrier, diluent or excipient US 8,921,361 B2 75 76 used will depend upon the means and purpose for which the route of administration, consistent with good medical prac compound of the present invention is being applied. Solvents tice. Factors for consideration in this context include the are generally selected based on Solvents recognized by per particular disorder being treated, the particular mammal sons skilled in the art as safe (GRAS) to be administered to a being treated, the clinical condition of the individual patient, mammal. In general, safe solvents are nontoxic aqueous sol the cause of the disorder, the site of delivery of the agent, the vents such as water and other non-toxic solvents that are method of administration, the scheduling of administration, soluble or miscible in water. Suitable aqueous solvents and other factors known to medical practitioners. The “thera include water, ethanol, propylene glycol, polyethylene gly peutically effective amount of the compound to be adminis cols (e.g., PEG 400, PEG 300), etc. and mixtures thereof. The tered will be governed by such considerations, and is the formulations may also include one or more buffers, stabiliz 10 minimum amount necessary to prevent, ameliorate, or treat ing agents, Surfactants, wetting agents, lubricating agents, the coagulation factor mediated disorder. Such amount is emulsifiers, Suspending agents, preservatives, antioxidants, preferably below the amount that is toxic to the host or ren opaquing agents, glidants, processing aids, colorants, Sweet ders the host significantly more Susceptible to bleeding. eners, perfuming agents, flavoring agents and other known As a general proposition, the initial pharmaceutically additives to provide an elegant presentation of the drug (i.e., 15 effective amount of the inhibitor administered parenterally a compound of the present invention or pharmaceutical com per dose will be in the range of about 0.01-100 mg/kg, namely position thereof) or aid in the manufacturing of the pharma about 0.1 to 20 mg/kg of patient body weight per day, with the ceutical product (i.e., medicament). typical initial range of compound used being 0.3 to 15 mg/kg/ The formulations may be prepared using conventional dis day. Solution and mixing procedures. For example, the bulk drug Acceptable diluents, carriers, excipients and stabilizers are Substance (i.e., compound of the present invention or stabi nontoxic to recipients at the dosages and concentrations lized form of the compound (e.g., complex with a cyclodex employed, and include buffers such as phosphate, citrate and trin derivative or other known complexation agent) is dis other organic acids; antioxidants including ascorbic acid and solved in a suitable solvent in the presence of one or more of methionine; preservatives (such as octadecyldimethylbenzyl the excipients described above. The compound of the present 25 ammonium chloride; hexamethonium chloride; benzalko invention is typically formulated into pharmaceutical dosage nium chloride, benzethonium chloride; phenol, butyl or ben forms to provide an easily controllable dosage of the drug and Zyl alcohol; alkyl parabens such as methyl or propyl paraben; to enable patient compliance with the prescribed regimen. catechol; resorcinol; cyclohexanol: 3-pentanol; and The pharmaceutical composition (or formulation) for m-cresol); low molecular weight (less than about 10 residues) application may be packaged in a variety of ways depending 30 polypeptides; proteins, such as serum albumin, gelatin, or upon the method used for administering the drug. Generally, immunoglobulins; hydrophilic polymers such as polyvi an article for distribution includes a container having depos nylpyrrolidone; amino acids such as glycine, glutamine, ited therein the pharmaceutical formulation in an appropriate asparagine, histidine, arginine, or lysine; monosaccharides, form. Suitable containers are well known to those skilled in disaccharides and other carbohydrates including glucose, the art and include materials such as bottles (plastic and 35 mannose, or dextrins; chelating agents such as EDTA: Sugars glass), Sachets, ampoules, plastic bags, metal cylinders, and Such as Sucrose, mannitol, trehalose or Sorbitol; salt-forming the like. The container may also include a tamper-proof counter-ions such as Sodium; metal complexes (e.g., Zn-pro assemblage to prevent indiscreet access to the contents of the tein complexes); and/or non-ionic Surfactants such as package. In addition, the container has deposited thereon a TWEENTM, PLURONICSTM or polyethylene glycol (PEG). label that describes the contents of the container. The label 40 The active pharmaceutical ingredients may also be entrapped may also include appropriate warnings. in microcapsules prepared, for example, by coacervation Pharmaceutical formulations of the compounds of the techniques or by interfacial polymerization, for example, present invention may be prepared for various routes and hydroxymethylcellulose or gelatin-microcapsules and poly types of administration. For example, a compound of formula (methylmethacylate) microcapsules, respectively, in colloi Ia-d having the desired degree of purity may optionally be 45 dal drug delivery systems (for example, liposomes, albumin mixed with pharmaceutically acceptable diluents, carriers, microspheres, microemulsions, nano-particles and nanocap excipients or stabilizers (Remington's Pharmaceutical Sci Sules) or in macroemulsions. Such techniques are disclosed in ences (1980) 16th edition, Osol, A. Ed.), in the form of a Remington’s Pharmaceutical Sciences 16th edition, Osol, A. lyophilized formulation, milled powder, or an aqueous solu Ed. (1980). tion, formulation may be conducted by mixing at ambient 50 Sustained-release preparations of compounds of formula temperature at the appropriate pH, and at the desired degree of Ia-d may be prepared. Suitable examples of Sustained-release purity, with physiologically acceptable carriers, i.e., carriers preparations include semipermeable matrices of solid hydro that are non-toxic to recipients at the dosages and concentra phobic polymers containing a compound of formula Ia-d. tions employed. The pH of the formulation depends mainly which matrices are in the form of shaped articles, e.g., films, on the particular use and the concentration of compound, but 55 or microcapsules. Examples of Sustained-release matrices may range from about 3 to about 8. Formulation in an acetate include polyesters, hydrogels (for example, poly(2-hydroxy buffer at pH 5 is a suitable embodiment. ethyl-methacrylate), or polyvinyl alcohol)), polylactides The compound of this invention for use herein is preferably (U.S. Pat. No. 3,773.919), copolymers of L-glutamic acid and sterile. In particular, formulations to be used for in vivo gamma-ethyl-L-glutamate, non-degradable ethylene-vinyl administration must be sterile. Such sterilization is readily 60 acetate, degradable lactic acid-glycolic acid copolymers such accomplished by filtration through sterile filtration mem as the LUPRON DEPOTTM (injectable microspheres com branes. posed of lactic acid-glycolic acid copolymer and leuprolide The compound ordinarily can be stored as a Solid compo acetate) and poly-D-(-)-3-hydroxybutyric acid. sition, a lyophilized formulation or as an aqueous solution. The formulations include those suitable for the administra The pharmaceutical compositions of the invention will be 65 tion routes detailed herein. The formulations may conve formulated, dosed and administered in a fashion, i.e., niently be presented in unit dosage form and may be prepared amounts, concentrations, schedules, course, vehicles and by any of the methods well known in the art of pharmacy. US 8,921,361 B2 77 78 Techniques and formulations generally are found in Reming While the phase may comprise merely an emulsifier, it desir ton's Pharmaceutical Sciences (Mack Publishing Co., Eas ably comprises a mixture of at least one emulsifier with a fat ton, Pa.). Such methods include the step of bringing into oran oil or with both a fat and an oil. Preferably, a hydrophilic association the active ingredient with the carrier which con emulsifier is included together with a lipophilic emulsifier stitutes one or more accessory ingredients. In general the which acts as a stabilizer. It is also preferred to include both an formulations are prepared by uniformly and intimately bring oil and a fat. Together, the emulsifier(s) with or without sta ing into association the active ingredient with liquid carriers bilizer(s) makeup the so-called emulsifying wax, and the wax or finely divided solid carriers or both, and then, if necessary, together with the oil and fat make up the so-called emulsify shaping the product. ingointment base which forms the oily dispersed phase of the Formulations of a compound of formula Ia-d suitable for 10 cream formulations. Emulsifiers and emulsion stabilizers oral administration may be prepared as discrete units such as suitable for use in the formulation of the invention include pills, capsules, cachets or tablets each containing a predeter Tween(R) 60, SpanR 80, cetostearyl alcohol, benzyl alcohol, mined amount of a compound of formula Ia-d. myristyl alcohol, glyceryl mono-stearate and sodium lauryl Compressed tablets may be prepared by compressing in a sulfate. Suitable machine the active ingredient in a free-flowing form 15 Aqueous Suspensions of formula Ia-d compounds contain Such as a powder or granules, optionally mixed with a binder, the active materials in admixture with excipients suitable for lubricant, inert diluent, preservative, Surface active or dis the manufacture of aqueous Suspensions. Such excipients persing agent. Molded tablets may be made by molding in a include a Suspending agent, such as Sodium carboxymethyl Suitable machine a mixture of the powdered active ingredient cellulose, croScarmellose, povidone, methylcellulose, moistened with an inert liquid diluent. The tablets may hydroxypropyl methylcellulose, sodium alginate, polyvi optionally be coated or scored and optionally are formulated nylpyrrolidone, gum tragacanth and gum acacia, and dispers So as to provide slow or controlled release of the active ingre ing or wetting agents such as a naturally occurring phos dient therefrom. phatide (e.g., lecithin), a condensation product of an alkylene Tablets, troches, lozenges, aqueous or oil suspensions, dis oxide with a fatty acid (e.g., polyoxyethylene Stearate), a persible powders or granules, emulsions, hard or soft cap 25 condensation product of ethylene oxide with a long chain Sules, e.g., gelatin capsules, syrups or elixirs may be prepared aliphatic alcohol (e.g., heptadecaethyleneoxycetanol), a con for oral use, formulations of compounds of formula Ia-d densation product of ethylene oxide with a partial ester intended for oral use may be prepared according to any derived from a fatty acid and a hexitol anhydride (e.g., poly method known to the art for the manufacture of pharmaceu oxyethylene Sorbitan monooleate). The aqueous Suspension tical compositions and Such compositions may contain one or 30 may also contain one or more preservatives such as ethyl or more agents including Sweetening agents, flavoring agents, n-propyl p-hydroxybenzoate, one or more coloring agents, coloring agents and preserving agents, in order to provide a one or more flavoring agents and one or more Sweetening palatable preparation. Tablets containing the active ingredient agents, such as Sucrose or saccharin. in admixture with non-toxic pharmaceutically acceptable The pharmaceutical compositions of compounds of the excipient which are suitable for manufacture of tablets are 35 invention may be in the form of a sterile injectable prepara acceptable. These excipients may be, for example, inert dilu tion, Such as a sterile injectable aqueous or oleaginous Sus ents, such as calcium or Sodium carbonate, lactose, calcium or pension. This Suspension may be formulated according to the Sodium phosphate, granulating and disintegrating agents, known art using those Suitable dispersing or wetting agents Such as maize starch, or alginic acid; binding agents, such as and Suspending agents which have been mentioned above. Starch, gelatin or acacia; and lubricating agents, such as mag 40 The sterile injectable preparation may also be a sterile inject nesium Stearate, Stearic acid or talc. Tablets may be uncoated able solution or Suspension in a non-toxic parenterally or may be coated by known techniques including microen acceptable diluent or solvent, such as a solution in 1,3-bu capsulation to delay disintegration and adsorption in the gas tanediol or prepared as a lyophilized powder. Among the trointestinal tract and thereby provide a Sustained action over acceptable vehicles and solvents that may be employed are a longer period. For example, a time delay material Such as 45 water, Ringer's Solution and isotonic sodium chloride solu glyceryl monostearate or glyceryl distearate alone or with a tion. In addition, sterile fixed oils may conventionally be wax may be employed. employed as a solvent or Suspending medium. For this pur For treatment of the eye or other external tissues, e.g., pose any bland fixed oil may be employed including synthetic mouth and skin, the formulations are preferably applied as a mono- or diglycerides. In addition, fatty acids such as oleic topical ointment or cream containing the active ingredient(s) 50 acid may likewise be used in the preparation of injectables. in an amount of, for example, 0.075 to 20% w/w. When The amount of active ingredient that may be combined formulated in an ointment, the active ingredients may be with the carrier material to produce a single dosage form will employed with either a paraffinic or a water-miscible oint vary depending upon the host treated and the particular mode ment base. Alternatively, the active ingredients may be for of administration. For example, a time-release formulation mulated in a cream with an oil-in-water cream base. 55 intended for oral administration to humans may contain If desired, the aqueous phase of the cream base may approximately 1 to 1000 mg of active material compounded include a polyhydric alcohol, i.e., an alcohol having two or with an appropriate and convenient amount of carrier material more hydroxyl groups such as propylene glycol, butane 1,3- which may vary from about 5 to about 95% of the total diol, mannitol, Sorbitol, glycerol and polyethylene glycol (in compositions (weight: weight). The pharmaceutical compo cluding PEG 400) and mixtures thereof. The topical formu 60 sition can be prepared to provide easily measurable amounts lations may desirably include a compound which enhances for administration. For example, an aqueous solution absorption or penetration of the active ingredient through the intended for intravenous infusion may contain from about 3 to skin or other affected areas. Examples of Such dermal pen 500 ug of the active ingredient per milliliter of solution in etration enhancers include dimethyl sulfoxide and related order that infusion of a suitable volume at a rate of about 30 analogs. 65 ni/hr can occur. The oily phase of the emulsions of this invention may be Formulations suitable for parenteral administration constituted from known ingredients in a known manner. include aqueous and nonaqueous sterile injection solutions US 8,921,361 B2 79 80 which may contain anti-oxidants, buffers, bacteriostats and erative properties or that is useful for treating a hyperprolif solutes which render the formulation isotonic with the blood erative disorder (e.g., cancer). The second compound of the of the intended recipient; and aqueous and non-aqueous ster pharmaceutical combination formulation or dosing regimen ille Suspensions which may include Suspending agents and preferably has complementary activities to the compound of thickening agents. formula Ia-d such that they do not adversely affect each other. Formulations suitable for topical administration to the eye Such compounds are suitably present in combination in also include eye drops wherein the active ingredient is dis amounts that are effective for the purpose intended. In one Solved or Suspended in a Suitable carrier, especially an aque embodiment, a composition of this invention comprises a ous solvent for the active ingredient. The active ingredient is compound of formula Ia-d, or a stereoisomer, geometric iso preferably present in Such formulations in a concentration of 10 mer, tautomer, solvate, metabolite, a N-oxide derivative or about 0.5 to 20% w/w, for example about 0.5 to 10% w/w, for pharmaceutically acceptable salt or prodrug thereof, in com example about 1.5% w/w. bination with a chemotherapeutic agent Such as described Formulations suitable for topical administration in the herein. mouth include lozenges comprising the active ingredient in a The combination therapy may be administered as a simul flavored basis, usually Sucrose and acacia or tragacanth; pas 15 taneous or sequential regimen. When administered sequen tilles comprising the active ingredient in an inert basis such as tially, the combination may be administered in two or more gelatin and glycerin, or Sucrose and acacia; and mouthwashes administrations. The combined administration includes coad comprising the active ingredient in a Suitable liquid carrier. ministration, using separate formulations or a single pharma Formulations for rectal administration may be presented as ceutical formulation, and consecutive administration in either a Suppository with a Suitable base comprising for example order, wherein preferably there is a time period while both (or cocoa butter or a salicylate. 00180 formulations suitable for all) active agents simultaneously exert their biological activi intrapulmonary or nasal administration have a particle size ties. for example in the range of 0.1 to 500 microns (including Suitable dosages for any of the above coadministered particle sizes in a range between 0.1 and 500 microns in agents are those presently used and may be lowered due to the increments microns such as 0.5. 1, 30 microns, 35 microns, 25 combined action (synergy) of the newly identified agent and etc.), which is administered by rapid inhalation through the other chemotherapeutic agents or treatments. nasal passage or by inhalation through the mouth So as to The combination therapy may provide “synergy and reach the alveolar sacs. Suitable formulations include aque prove “synergistic”, i.e., the effect achieved when the active ous or oily Solutions of the active ingredient. Formulations ingredients used together is greater than the Sum of the effects suitable for aerosol or dry powder administration may be 30 that results from using the compounds separately. A syner prepared according to conventional methods and may be gistic effect may be attained when the active ingredients are: delivered with other therapeutic agents such as compounds (1) co-formulated and administered or delivered simulta heretofore used in the treatment or prophylaxis disorders as neously in a combined, unit dosage formulation; (2) delivered described below. by alternation or in parallel as separate formulations; or (3) by Formulations suitable for vaginal administration may be 35 some other regimen. When delivered in alternation therapy, a presented as pessaries, tampons, creams, gels, pastes, foams synergistic effect may be attained when the compounds are or spray formulations containing in addition to the active administered or delivered sequentially, e.g., by different ingredient such carriers as are known in the art to be appro injections in separate Syringes, separate pills or capsules, or priate. separate infusions. In general, during alternation therapy, an The formulations may be packaged in unit-dose or multi 40 effective dosage of each active ingredient is administered dose containers, for example sealed ampoules and vials, and sequentially, i.e., serially, whereas in combination therapy, may be stored in a freeze-dried (lyophilized) condition requir effective dosages of two or more active ingredients are ing only the addition of the sterile liquid carrier, for example administered together. water, for injection immediately prior to use. Extemporane In a particular embodiment of anti-cancer therapy, a com ous injection solutions and Suspensions are prepared from 45 pound of formula Ia-d, or a stereoisomer, geometric isomer, sterile powders, granules and tablets of the kind previously tautomer, solvate, metabolite, N-oxide derivative or pharma described. Preferred unit dosage formulations are those con ceutically acceptable salt or prodrug thereof, may be com taining a daily dose or unit daily Sub-dose, as herein above bined with other chemotherapeutic, hormonal or antibody recited, or an appropriate fraction thereof, of the active ingre agents such as those described herein, as well as combined dient. 50 with Surgical therapy and radiotherapy. Combination thera The invention further provides veterinary compositions pies according to the present invention thus comprise the comprising at least one active ingredient as above defined administration of at least one compound of formula Ia-d, or a together with a veterinary carrier therefore. Veterinary carri Stereoisomer, geometric isomer, tautomer, Solvate, metabo ers are materials useful for the purpose of administering the lite, N-oxide derivative orpharmaceutically acceptable salt or composition and may be solid, liquid or gaseous materials 55 prodrug thereof, and the use of at least one other cancer which are otherwise inert or acceptable in the veterinary art treatment method. The amounts of the compound(s) of for and are compatible with the active ingredient. These veteri mula Ia-d and the other pharmaceutically active chemothera nary compositions may be administered parenterally, orally peutic agent(s) and the relative timings of administration will or by any other desired route. be selected in order to achieve the desired combined thera Combination Therapy 60 peutic effect. The compounds of the invention may be employed alone or Metabolites of Compounds of the Invention in combination with other therapeutic agents for the treatment Also falling within the scope of this invention are the in of a disease or disorder described herein, such as a hyperpro vivo metabolic products of formulas (I) and (Ia) to (Ii) liferative disorder (e.g., cancer). In certain embodiments, a described herein. Such products may result for example from compound of formula Ia-d is combined in a pharmaceutical 65 the oxidation, reduction, hydrolysis, amidation, deamidation, combination formulation, or dosing regimen as combination esterification, deesterification, enzymatic cleavage, and the therapy, with a second compound that has anti-hyperprolif like, of the administered compound. Accordingly, the inven US 8,921,361 B2 81 82 tion includes metabolites of compounds of formulas (I) and each C-aminoacyl group is independently selected from the (Ia) to (Ii), including compounds produced by a process com naturally occurring L-amino acids, P(O)(OH), —P(O)(O prising contacting a compound of this invention with a mam (C-C)alkyl) or glycosyl (the radical resulting from the mal for a period of time sufficient to yield a metabolic product removal of a hydroxyl group of the hemiacetal form of a thereof. carbohydrate). O0196. For additional examples of prodrug Metabolite products typically are identified by preparing a derivatives, see, for example, a) Design of Prodrugs, edited by radiolabeled (e.g., C or H) isotope of a compound of the H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, invention, administering it parenterally in a detectable dose Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic (e.g., greater than about 0.5 mg/kg) to an animal such as rat, Press, 1985); b) A Textbook of Drug Design and Develop mouse, guinea pig, monkey, or to man, allowing Sufficient 10 ment, edited by Krogsgaard-Larsen and H. Bundgaard, Chap time for metabolism to occur (typically about 30 seconds to 30hours) and isolating its conversion products from the urine, ter5 "Design and Application of Prodrugs.” by H. Bundgaard blood or other biological samples. These products are easily p. 113-191 (1991); c) H. Bundgaard, Advanced Drug Deliv isolated since they are labeled (others are isolated by the use ery Reviews, 8:1-38 (1992); d) H. Bundgaard, et al., Journal of antibodies capable of binding epitopes Surviving in the 15 of Pharmaceutical Sciences, 77:285 (1988); and e) N. metabolite). The metabolite structures are determined in con Kakeya, et al., Chem. Pharm. Bull., 32:692 (1984), each of ventional fashion, e.g., by MS, LC/MS or NMR analysis. In which is specifically incorporated herein by reference. general, analysis of metabolites is done in the same way as Articles of Manufacture conventional drug metabolism studies well known to those In another embodiment of the invention, an article of skilled in the art. The metabolite products, so long as they are manufacture, or "kit, containing materials useful for the not otherwise found in Vivo, are useful in diagnostic assays treatment of the diseases and disorders described above is for therapeutic dosing of the compounds of the invention. provided. In one embodiment, the kit comprises a container Prodrugs of the Compounds of the Invention comprising a compound of formula Ia-d, or a stereoisomer, In addition to compounds of the invention, the invention geometric isomer, tautomer, Solvate, metabolite, N-oxide also includes pharmaceutically acceptable prodrugs of Such 25 derivative or pharmaceutically acceptable salt or prodrug compounds. Prodrugs include compounds wherein an amino thereof. The kit may further comprise a label or package insert acid residue, or a polypeptide chain of two or more (e.g., two, on or associated with the container. The term "package insert three or four) amino acid residues, is covalently joined is used to refer to instructions customarily included in com through an amide or ester bond to a free amino, hydroxy or mercial packages of therapeutic products, that contain infor carboxylic acid group of a compound of the present invention. 30 mation about the indications, usage, dosage, administration, The amino acid residues include but are not limited to the 20 contraindications and/or warnings concerning the use of Such naturally occurring amino acids commonly designated by therapeutic products. Suitable containers include, for three letter symbols and also includes phosphoserine, phos example, bottles, vials, Syringes, blister pack, etc. The con phothreonine, phosphotyrosine, 4-hydroxyproline, hydroxy tainer may beformed from a variety of materials such as glass lysine, demosine, isodemosine, gamma-carboxyglutamate, 35 or plastic. The container may hold a compound of formula hippuric acid, octahydroindole-2-carboxylic acid, statine, Ia-dora formulation thereof which is effective for treating the 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, penicil condition and may have a sterile access port (for example, the lamine, ornithine, 3-methylhistidine, norvaline, beta-alanine, container may be an intravenous solution bag or a vial having gamma-aminobutyric acid, citrulline, homocysteine, a stopper pierceable by a hypodermic injection needle). At homoserine, methyl-alanine, para-benzoylphenylalanine, 40 least one active agent in the composition is a compound of phenylglycine, propargylglycine, sarcosine, methionine Sul formula Ia-d. The label or package insert indicates that fone and tert-butylglycine. the composition is used for treating the condition of choice, Additional types of prodrugs are also encompassed. For Such as cancer. In addition, the label or package insert may instance, a free carboxyl group of a compound of the inven indicate that the patient to be treated is one having a disorder tion can be derivatized as an amide or alkyl ester. As another 45 Such as a hyperproliferative disorder, neurodegeneration, car example, compounds of this invention comprising free diac hypertrophy, pain, migraine or a neurotraumatic disease hydroxy groups may be derivatized as prodrugs by converting or event. In one embodiment, the label or package inserts the hydroxy group into a group Such as, but not limited to, a indicates that the composition comprising a compound of phosphate ester, hemisuccinate, dimethylaminoacetate, or formula Ia-d can be used to treat a disorder resulting from phosphoryloxymethyloxycarbonyl group, as outlined in 50 abnormal cell growth. The label or package insert may also Advanced Drug Delivery Reviews, (1996) 19:115. Carbam indicate that the composition can be used to treat other dis ate prodrugs of hydroxy and amino groups are also included, orders. Alternatively, or additionally, the article of manufac as are carbonate prodrugs, sulfonate esters and Sulfate esters ture may further comprise a second container comprising a of hydroxy groups. Derivatization of hydroxy groups as (acy pharmaceutically acceptable buffer, Such as bacteriostatic loxy)methyl and (acyloxy)ethyl ethers, wherein the acyl 55 water for injection (BWFI), phosphate-buffered saline, Ring group may be an alkyl ester optionally Substituted with er's solution and dextrose solution. It may further include groups including, but not limited to, ether, amine and car other materials desirable from a commercial and user stand boxylic acid functionalities, or where the acyl group is an point, including other buffers, diluents, filters, needles, and amino acid ester as described above, are also encompassed. Syringes. Prodrugs of this type are described in J. Med. Chem. (1996), 60 The kit may further comprise directions for the adminis 39:10. More specific examples include replacement of the tration of the compound of formula Ia-d and, if present, the hydrogen atom of the alcohol group with a group Such as second pharmaceutical formulation. For example, if the kit (C-C)alkanoyloxymethyl, 1-((Ci-C)alkanoyloxy)ethyl, comprises a first composition comprising a compound of 1-methyl-1-((C-C)alkanoyloxy)ethyl, (Ci-C)alkoxycar formula Ia-danda second pharmaceutical formulation, the kit bonyloxymethyl, N-(C-C)alkoxycarbonylaminomethyl, 65 may further comprise directions for the simultaneous, Succinoyl (CrCl)alkanoyl, C-amino(C-C)alkanoyl, aryla sequential or separate administration of the first and second cyl and C-aminoacyl, or C.-aminoacyl-C-aminoacyl, where pharmaceutical compositions to a patient in need thereof. US 8,921,361 B2 83 84 In another embodiment, the kits are suitable for the deliv COR) were applied and plates were scanned on an Odyssey ery of solid oral forms of a compound of formula Ia-d, such as reader to detect pPKB/PKB ratios. tablets or capsules. Such a kit preferably includes a number of The following compounds have shown particularly inter unit dosages. Such kits can include a card having the dosages esting biological activities: oriented in the order of their intended use. An example of such 4,4'-(6-(pyridin-3-yl)-1,3,5-triazine-2,4-diyl)dimorpholine, a kit is a “blister pack”. Blister packs are well known in the 3-(4,6-dimorpholino-1,3,5-triazin-2-yl)phenol, packaging industry and are widely used for packaging phar 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)pyridin-3-ol, maceutical unit dosage forms. If desired, a memory aid can be 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)pyridin-2-amine, provided, for example in the form of numbers, letters, or other 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)pyrimidin-2-amine, 10 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)-4-(trifluoromethyl) markings or with a calendarinsert, designating the days in the pyridin-2-amine, treatment schedule in which the dosages can be administered. 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)-4-(trifluoromethyl) According to one embodiment, a kit may comprise (a) a pyrimidin-2-amine, first container with a compound of formula Ia-d contained N-(5-(4,6-dimorpholino-1,3,5-triazin-2-yl)pyridin-2-yl)ac therein; and optionally (b) a second container with a second 15 etamide, pharmaceutical formulation contained therein, wherein the N-(5-(4,6-dimorpholino-1,3,5-triazin-2-yl)pyrimidin-2-yl) second pharmaceutical formulation comprises a second com acetamide, pound with anti-hyperproliferative activity. Alternatively, or N-(5-(4,6-dimorpholino-1,3,5-triazin-2-yl)-4-(trifluorom additionally, the kit may further comprise a third container ethyl)pyrimidin-2-yl)acetamide, comprising a pharmaceutically-acceptable buffer, such as N-(5-(4,6-dimorpholino-1,3,5-triazin-2-yl)-4-(trifluorom bacteriostatic water for injection (BWFI), phosphate-buff ethyl)pyridin-2-yl)acetamide, ered Saline, Ringer's Solution and dextrose Solution. It may 5-(4,6-dimorpholino-1,3,5-triazin-2-yl)-6-(trifluoromethyl) further include other materials desirable from a commercial pyridin-2-amine, and user standpoint, including other buffers, diluents, filters, 4,4'-(6-(1H-indazol-5-yl)-1,3,5-triazine-2,4-diyl)dimorpho needles, and Syringes. 25 line, In certain other embodiments wherein the kit comprises a 4,4'-(6-(1H-indol-5-yl)-1,3,5-triazine-2,4-diyl)dimorpho composition of formula Ia-d and a second therapeutic agent, line, the kit may comprise a container for containing the separate 4,4'-(6-(1H-benzodimidazol-5-yl)-1,3,5-triazine-2,4-diyl) compositions such as a divided bottle or a divided foil packet, dimorpholine, however, the separate compositions may also be contained 30 4,4'-(6-(1H-benzodimidazol-4-yl)-1,3,5-triazine-2,4-diyl) within a single, undivided container. Typically, the kit com dimorpholine, prises directions for the administration of the separate com 4,4'-(6-(1H-indol-4-yl)-1,3,5-triazine-2,4-diyl)dimorpho ponents. The kit form is particularly advantageous when the line, separate components are preferably administered in different 4,4'-(6-(1H-indazol-4-yl)-1,3,5-triazine-2,4-diyl)dimorpho dosage forms (e.g., oral and parenteral), are administered at 35 line, different dosage intervals, or when titration of the individual 4,4'-(6-(1H-indazol-6-yl)-1,3,5-triazine-2,4-diyl)dimorpho components of the combination is desired by the prescribing line, physician. 5-(4-morpholino-6-(pyridin-2-ylmethoxy)-1,3,5-triazin-2- Example of Biological Evaluation yl)pyridin-2-amine, Determination of the potential to target PI3K/PI3K-related 40 5-(4-morpholino-6-(pyridin-2-ylmethoxy)-1,3,5-triazin-2- kinases (PIKK) of a compound of formula I is possible by a yl)pyrimidin-2-amine, number of direct and indirect detection methods. Certain 4-(4-(1H-indol-4-yl)-6-(pyridin-2-ylmethoxy)-1,3,5-triazin exemplary compounds described herein were assayed for 2-yl)morpholine, their phospho PKB blocking activity and their invitro activity 4-(4-(1H-indazol-4-yl)-6-(pyridin-2-ylmethoxy)-1,3,5-tri against tumor cells. The range of phospho PKB activities was 45 azin-2-yl)morpholine, less than 1 nM (nanomolar) to about 10 uM (micromolar). 4-(4-(1H-benzodimidazol-4-yl)-6-(pyridin-2-ylmethoxy)- Other exemplary compounds of the invention had phospho 1,3,5-triazin-2-yl)morpholine, PKB blocking activity ICs values less than 10 nM. Certain 4-(4-(1H-benzodimidazol-5-yl)-6-(pyridin-2-ylmethoxy)- compounds of the invention had tumor cell-based activity 1,3,5-triazin-2-yl)morpholine, ICso values less than 100 nM. 50 4-(4-(1H-indazol-5-yl)-6-(pyridin-2-ylmethoxy)-1,3,5-tri The cytotoxic or cytostatic activity of formula I exemplary azin-2-yl)morpholine, compounds was measured by: establishing a proliferating 4-(4-(1H-indol-5-yl)-6-(pyridin-2-ylmethoxy)-1,3,5-triazin mammalian tumor cell lines in a cell culture medium, adding 2-yl)morpholine, a formula I compound, culturing the cells for a period from 3-(4-morpholino-6-(pyridin-2-ylmethylamino)-1,3,5-tri about 6 hours to about 3 days; and measuring cell viability. 55 azin-2-yl)phenol, Cell-based assays were used to measure viability, i.e. prolif 4-(6-aminopyridin-3-yl)-6-morpholino-N-(pyridin-2-ylm eration (ICs), cytotoxicity (ECs), and induction of apopto ethyl)-1,3,5-triazin-2-amine, sis (caspase activation). 4-(2-aminopyrimidin-5-yl)-6-morpholino-N-(pyridin-2-yl The in vitro potency of formula I compounds was mea methyl)-1,3,5-triazin-2-amine, sured by the in-cell Western assay designed in laboratories at 60 4-(1H-indol-5-yl)-6-morpholino-N-(pyridin-2-ylmethyl)-1, University of Basel. This assay method was conducted in 3.5-triazin-2-amine, microtiter plate formats, making it amenable to high-through 4-(1H-indazol-5-yl)-6-morpholino-N-(pyridin-2-ylmethyl)- put screening (HTS). Inhibitors were added to the medium 1.3.5-triazin-2-amine, and incubated. Antibodies diluted in PBS/T against pPKB 4-(1H-benzodimidazol-5-yl)-6-morpholino-N-(pyridin-2- Ser473 (Cell Signalling) and PKB (gift from E. Hirsch) or 65 ylmethyl)-1,3,5-triazin-2-amine, pS6 Ser 235/236 (Cell Signalling) were incubated overnight 4-(1H-benzodimidazol-4-yl)-6-morpholino-N-(pyridin-2- and then secondary fluorescently labelled antibodies (LI ylmethyl)-1,3,5-triazin-2-amine,

US 8,921,361 B2 141 142 TABLE 1-continued

Cpd No. Structure Name

89 Bodipy = Tag molecule

90 Bodipy = Tag molecule

ON

NN CF --- N 2 N