US 2011 02881 63A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0288163 A1 Fraser et al. (43) Pub. Date: Nov. 24, 2011
(54) METHODS OF USING MACROCYCLC 60/622,005, filed on Oct. 27, 2004, provisional appli MODULATORS OF THE GHRELIN cation No. 60/642,271, filed on Jan. 7, 2005. RECEPTOR Publication Classification (76) Inventors: Graeme L. Fraser, Quebec (CA); (51) Int. Cl. Hamid R. Hoveyda, Quebec (CA); A 6LX 3L/395 (2006.01) Mark L. Peterson, Quebec (CA) A6IP 43/00 (2006.01) (21) Appl. No.: 13/173,929 (52) U.S. Cl...... S14/450 (57) ABSTRACT (22) Filed: Jun. 30, 2011 The present invention provides novel conformationally-de fined macrocyclic compounds that have been demonstrated to Related U.S. Application Data be selective modulators of the ghrelin receptor (growth hor (60) Division of application No. 12/333,026, filed on Dec. mone secretagogue receptor, GHS-R1a and Subtypes, iso 11, 2008, which is a continuation of application No. forms and variants thereof). Methods of synthesizing the 1 1/149,512, filed on Jun. 10, 2005, now Pat. No. 7,491, novel compounds are also described herein. These com 695, which is a continuation-in-part of application No. pounds are useful as agonists of the ghrelin receptor and as 10/872,142, filed on Jun. 18, 2004, now Pat. No. 7,521, medicaments for treatment and prevention of a range of medi 420. cal conditions including, but not limited to, metabolic and/or endocrine disorders, gastrointestinal disorders, cardiovascu (60) Provisional application No. 60/479,223, filed on Jun. lar disorders, obesity and obesity-associated disorders, cen 18, 2003, provisional application No. 60/621,642. tral nervous system disorders, genetic disorders, hyperprolif filed on Oct. 26, 2004, provisional application No. erative disorders and inflammatory disorders. Patent Application Publication Nov. 24, 2011 Sheet 1 of 21 US 2011/02881 63 A1
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Target; h(GHS-R Radioligand; 125Ghrelin
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f Ki = 0.34 nM Compound 25
LOg M of compound s FIGURE 4E
Compound 1
-10 -9 -8 -7 -6 -5 -4 Log Concentration (M) FIGURE 5A Patent Application Publication Nov. 24, 2011 Sheet 7 of 21 US 2011/02881 63 A1
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METHODS OF USING MACROCYCLC and skeletal muscles. This receptor was identified and cloned MODULATORS OF THE GHRELIN prior to the isolation and characterization of the endogenous RECEPTOR peptide ligand and is distinct from other receptors involved in the regulation of growth hormone (GH) secretion, in particu RELATED APPLICATION INFORMATION lar, the growth hormone-releasing hormone (GHRH) recep 0001. This application is a continuation application of tOr. U.S. patent application Ser. No. 1 1/149.512, filed Jun. 10, 0004. A unique characteristic of both the rat and human 2005, which is a continuation-in-part under 35 U.S.C. S 120 of peptides is the presence of the n-octanoyl (Oct) moiety on U.S. patent application Ser. No. 10/872,142, filed Jun. 18. Ser. However, the des-acyl form predominates in circula 2004, currently pending, which claims the benefit under 35 tion, with approximately 90% of the hormone in this form. U.S.C. S119(e) of U.S. Provisional Patent Application Ser. This group is derived from a post-translational modification No. 60/479,223, filed Jun. 18, 2003. The continuation-in-part and appears relevant for bioactivity and possibly also for application also claims the benefit under 35 U.S.C. S 119(e) of transport into the CNS. (Banks, W.A.: Tschöp, M.: Robinson, U.S. Provisional Patent Application Ser. No. 60/621,642, S. M.: Heiman, M. L. Extent and direction of ghrelin transport filed Oct. 26, 2004, U.S. Provisional Patent Application Ser. across the blood-brain barrier is determined by its unique No. 60/622,055, filed Oct. 27, 2004, and U.S. Provisional primary structure. J. Pharmacol. Exp. Ther: 2002, 302, 822 Patent Application Ser. No. 60/642,271, filed Jan. 7, 2005. 827.) In a GH-releasing assay, the des-octanoyl form of the The disclosures of the above-referenced applications are hormone was at least 100-fold less potent than the parent incorporated herein by reference in their entireties. peptide, although it has been Suggested that the des-acyl species may be responsible for some of the other biological FIELD OF THE INVENTION effects associated with ghrelin. This des-acyl form has also been postulated to be primarily responsible for the cardiovas 0002 The present invention relates to novel conforma cular and cell proliferation effects attributed to ghrelin, while tionally-defined macrocyclic compounds that bind to and/or the acylated form participates in maintenance of energy bal are functional modulators of the ghrelin (growth hormone ance and growth hormone release. (Baldanzi, G.; Filigh secretagogue) receptor including GHS-R1a and Subtypes, enddu, N.; Cutrupi, S.; et al. Ghrelin and des-acyl ghrelin isoforms and/or variants thereof. The present invention also inhibit cell death in cardiomyocytes and endothelial cells relates to intermediates of these compounds, pharmaceutical through ERK1/2 and PI-3 kinase/AKT. J. Cell Biol. 2002, compositions containing these compounds and methods of 159, 1029-1037) Similarly, des-Gln-ghrelin and its using the compounds. These novel macrocyclic compounds octanoylated derivative have been isolated as endogenous are useful as therapeutics for a range of disease indications. In forms of the hormone arising from alternative splicing of the particular, these compounds are useful for treatment and pre ghrelin gene, but both are found to be inactive in stimulating vention of gastrointestinal disorders including, but not lim GH release in vivo. (Hosoda, H.; Kojima, M.; Matsuo, H.; ited to, post-operative ileus, gastroparesis, including diabetic Kangawa, K. Purification and characterization of rat des gastroparesis, opioid bowel dysfunction, chronic intestinal Gln-ghrelin, a second endogenous ligand for the growth pseudo-obstruction, short bowel syndrome and functional hormone secretagogue receptor. J. Biol. Chem. 2000, 275, gastrointestinal disorders. 21995-2120.). Other minor forms of ghrelin produced by post-translational processing have been observed in plasma, BACKGROUND OF THE INVENTION although no specific activity has been attributed to them. 0003. The improved understanding of various physiologi (Hosoda, H.; Kojima, M.: et al. Structural divergence of cal regulatory pathways enabled through the research efforts human ghrelin. Identification of multiple ghrelin-derived in genomics and proteomics has begun to impact the discov molecules produced by post-translational processing. J. Biol. ery of novel pharmaceutical agents. In particular, the identi Chem. 2003, 278, 64-70.) fication of key receptors and their endogenous ligands has 0005. Even prior to the isolation of this receptor and its created new opportunities for exploitation of these receptor/ endogenous peptide ligand, a significant amount of research ligand pairs as therapeutic targets. For example, ghrelin is a was devoted to finding agents that can stimulate GH secre recently characterized 28-amino acid peptide hormone iso tion. The proper regulation of human GH has significance not lated originally from the stomach of rats with the orthologue only for proper body growth, but also a range of other critical Subsequently identified in humans. (Kojima, M., Hosoda, H. physiological effects. GH and other GH-stimulating peptides, et al. Nature 1999, 402, 656-660.) The existence of this pep such as GHRH and growth hormone releasing factor (GRF), tide in a range of other species suggests a conserved and as well as their derivatives and analogues, are administered important role in normal body function. This peptide has been via injection. Therefore, to better take advantage of these demonstrated to be the endogenous ligand for a previously positive effects, attention was focused on the development of orphan G protein-coupled receptor (GPCR), type 1 growth orally active therapeutic agents that would increase GH secre hormone secretatogue receptor (hGHS-R1a) (Howard, A. D.; tion, termed GH secretagogues (GHS). Additionally, use of Feighner, S. D.; et al. A receptor in pituitary and hypothala these agents was expected to more closely mimic the pulsatile mus that functions in growth hormone release. Science 1996, physiological release of GH. 273, 974-977.) found predominantly in the brain (arcuate 0006 Beginning with the identification of the growth hor nucleus and Ventromedial nucleus in the hypothalamus, hip mone-releasing peptides (GHRP) in the late 1970's, (Bowers, pocampus and Substantia nigra) and pituitary. (U.S. Pat. No. C. Y. Growth hormone-releasing peptides: physiology and 6.242, 199; Intl. Pat. Appl. Nos. WO 97/21730 and WO clinical applications. Curr. Opin. Endocrinol. Diabetes 2000, 97/22004) The receptor has also been detected in other areas 7, 168-174; Camanni, F. Ghigo, E.; Arvat, E. Growth hor of the central nervous system (CNS) and in peripheral tissues, mone-releasing peptides and their analogs. Front. Neurosci. for instance adrenal and thyroid glands, heart, lung, kidney, 1998, 19, 47-72; Locatelli, V., Torsello, A. Growth hormone US 2011/02881 63 A1 Nov. 24, 2011 secretagogues: focus on the growth hormone-releasing pep leptin. Indeed, it was the first gut peptide proven to have such tides. Pharmacol. Res. 1997, 36, 415-423.) a host of agents orexigenic properties. (Kojima, M., Kangawa, K. Ghrelin, an have been studied for their potential to act as GHS. In addition orexigenic signaling molecule from the gastrointestinal tract. to their stimulation of GH release and concomitant positive Curr. Opin. Pharmacology 2002, 2, 665-668.) The hormone effects in that regard, GHS were projected to have utility in also is implicated in the hypothalamic regulation of the Syn the treatment of a variety of other disorders, including wast thesis and secretion of a number of other neuropeptides ing conditions (cachexia) as seen in HIV patients and cancer involved in appetite and feeding behavior. Levels of ghrelin induced anorexia, musculoskeletal frailty in the elderly, and are elevated in response to fasting or extended food restric growth hormone deficient diseases. Many efforts over the tion. (Nakazato, M.; Murakami, N.; Date, Y.: Kojima, M.: et past 25 years have yielded a number of potent, orally available al. A role for ghrelin in the central regulation of feeding. GHS. (Smith, R. G.; Sun, Y. X., Beatancourt, L.; Asnicar, M. Nature 2001, 409, 194-198) For example, subjects suffering Growth hormone secretagogues: prospects and pitfalls. Best with anorexia or bulimia exhibit elevated ghrelin levels. Cir Pract. Res. Clin. Endocrinol. Metab. 2004, 18, 333-347: culating levels of the hormone have been found to rise before Fehrentz, J.-A.; Martinez, J.; Boeglin, D.; Guerlavais, V.: meals and fall after meals. In addition, diet-induced weight Deghenghi, R. Growth hormone secretagogues: Past, present loss leads to increased ghrelin levels, although obese Subjects and future. IDrugs 2002, 5,804-814; Svensson, J. Exp. Opin. who have gastric bypass Surgery do not likewise experience Ther. Patents 2000, 10, 1071-1080; Nargund, R. P.; Patchett, Such an increase. (Cummings, D. E.; Weigle, D. S.; Frayo, R. A. A. et al. Peptidomimetic growth hormone secretagogues. S.; et al. Plasmaghrelin levels after diet-induced weight loss Design considerations and therapeutic potential. J. Med. or gastric bypass surgery. N. Engl. J. Med. 2002, 346, 1623 Chem. 1998, 41,3103-3127: Ghigo, E: Arvat, E.: Camanni, F. 1630) Orally active growth hormone secretagogues: state of the art 0008. This intimate involvement of ghrelin in control of and clinical perspective. Ann. Med. 1998, 30, 159-168; Smith, food intake and appetite has made it an attractive target for R. G.; Vander Ploeg, L. H. T.; Howard, A. D.; Feighner, S.D.: obesity research. Indeed, few other natural substances have et al. Peptidomimetic regulation of growth hormone secre been demonstrated to be involved in the modulation of both tion. Endocr: Rev. 1997, 18, 621-645.) These include small GH secretion and food intake. peptides, such as hexarelin (Zentaris) and ipamorelin (Novo 0009. An additional effect of ghrelin that has not to date Nordisk), and adenosine analogues, as well as Small mol been exploited for therapeutic purposes is in modulating gas ecules such as carpomorelin (Pfizer), L-252,564 (Merck), tric motility and gastric acid secretion. The prokinetic activity MK-0677 (Merck), NN703 (Novo Nordisk), G-7203 (Genen appears to be independent of the GH-secretory action and is tech), S-37435 (Kaken) and SM-130868 (Sumitomo), likely mediated by the Vagal-cholinergic muscarinic pathway. designed to be orally active for the stimulation of growth The dose levels required are equivalent to those necessary for hormone. However, clinical testing with Such agents have the hormone's GH and appetite stimulation actions. It is note rendered disappointing results due to, among other things, worthy that, in contrast to its inactivity for ghrelin's other lack of efficacy over prolonged treatment or undesired side actions, the des-Gln'' peptide demonstrated promotion of effects, including irreversible inhibition of cytochrome P450 motility as well. (Trudel, L.; Bouin, M.; Tomasetto, C. Eber enzymes (Zdravkovic M. Olse, A. K. Christiansen, T.; et al. ling, P.; St-Pierre, S.; Bannon, P.; L'Heureux, M. C.; Poitras, Eur: J. Clin. Pharmacol. 2003, 58,683-688.) Therefore, there P. Two new peptides to improve post-operative gastric ileus in remains a need for pharmacological agents that could effec dog. Peptides 2003, 24, 531-534; Trudel, L.; Tomasetto, C.; tively target the GHS-R1a receptor for therapeutic action. Rio, M. C.; Bouin, M.; Plourde, V.: Eberling, P: Poitras, P. 0007. Despite its involvement in GH modulation, ghrelin Ghrelin/motilin-related peptide is a potent prokinetic to is primarily synthesized in the oxyntic gland of the stomach, reverse gastric postoperative ileus in rats. Am. J. Physiol. although it is also produced in lesser amounts in other organs, 2002, 282, G948-G952; Peeters, T. L. Central and peripheral including the kidney, pancreas and hypothalamus. (Kojima, mechanisms by which ghrelin regulates gut motility. J. M.; Hsoda, H. Kangawa, K. Purification and distribution of Physiol. Pharmacol. 2003, 54 (Supp. 4), 95-103.) ghrelin: the natural endogenous ligand for the growth hor 0010 Ghrelin also has been implicated in various aspects mone secretagogue receptor. Horm. Res. 2001, 56 (Suppl. 1), of reproduction and neonatal development. (Arvat, E., Gian 93-97: Ariyasu, H.; Takaya, K.; Tagami. T.; et al. Stomach is otti, L., Giordano, R., et al. Growth hormone-releasing hor a major source of circulating ghrelin, and feeding State deter mone and growth hormone secretagogue-receptor ligands. mines plasma ghrelin-like immunoreactivity levels in Focus on reproductive system. Endocrine 2001, 14, 35-43) humans. J. Clin. Endocrinol. Metab. 2001, 86, 4753-4758) In Also of significance are the cardiovascular effects of ghrelin, addition to its role in stimulating GH release, the hormone has since the peptide is a powerful vasodilator. As such, ghrelin a variety of other endocrine and non-endocrine functions agonists have potential for the treatment of chronic heart (Broglio, F. Gottero, C.; Arvat, E.: Ghigo, E. Endocrine and failure (Nagaya, N.; Kangawa, K. Ghrelin, a novel growth non-endocrine actions of ghrelin. Horm. Res. 2003, 59, 109 hormone-releasing peptide, in the treatment of chronic heart 117) and has been shown to interact with a number of other failure. Regul. Pept. 2003, 114, 71-77; Nagaya, N.; Kangawa, systems in playing a role in maintaining proper energy bal K. Ghrelin improves left ventricular dysfunction and cardiac ance. (Horvath, T. L.; Diano, S.; Sotonyi, P.; Heiman, M.: cachexia in heart failure. Curr: Opin. Pharmacol. 2003, 3, Tschöp, M. Ghrelin and the regulation of energy balance—a 146-151; Bedendi, I., Alloatti, G.; Marcantoni, A.; Malan, D.; hypothalamic perspective. Endocrinology 2001, 142, 4163 Catapano, F: Ghé, C.; et al. Cardiac effects of ghrelin and its 4169; Casanueva, F. F.; Dieguez, C. Ghrelin: the link con endogenous derivatives des-octanoyl ghrelin and des-Gln necting growth with metabolism and energy homeostasis. ghrelin. Eur: J. Pharmacol. 2003,476, 87-95) Intl. Pat. Appl. Rev. Endocrinol. Metab. Disord. 2002, 3, 325-338). In par Publ. WO 2004/014412 describes the use of ghrelin agonists ticular, ghrelin plays a role as an orexigenic signal in the for the protection of cell death in myocardial cells and as a control offeeding, in which it acts to counteract the effects of cardioprotectant treatment for conditions leading to heart US 2011/02881 63 A1 Nov. 24, 2011 failure. Lastly, evidence has been obtained that ghrelin may N.: Escher. F.; Collu, R.; Deghenghi, R.: Locatelli, V. Ghigo, have implications in anxiety and other CNS disorders as well E.; Muccioli, G.; Boghen, M.: Nilsson, M. Endocrinology as the improvement of memory. (Carlini, V. P. Monzon, M. 1998, 139, 432-435.) Differences between overall GHS-R E. Varas, M. M., Cragnolini, A. B., Schioth, H. B., Sci expression and that of the GHS-R1a subtype in rat testis have monelli, T.N. de Barioglio, S. R. Ghrelin increases anxiety been reported. (Barreiro, M. L.; Suominen, J.S.: Gaytan, F.; like behavior and memory retention in rats. Biochem. Bio Pinilla, L.; Chopin, L. K. Casanueva, F. F.; Dieguez, C.; phys. Res. Commun. 2002, 299,739-743) Aguilar, E., Toppari, J.; Tena-Sempere, M. Developmental, 0011. The myriad effects of ghrelin in humans have sug stage-specific, and hormonally regulated expression of gested the existence of Subtypes for its receptor, although growth hormone secretagogue receptor messenger RNA in none have as yet been identified. (Torsello, A.; Locatelli, Y.: rattestis. Biol. Reproduction 2003, 68, 1631-1640) A GHS-R Melis, M. R.: Succu, S.; Spano, M.S.: Deghenghi, R.; Muller, Subtype on cholinergic nerves is postulated as an explanation E. E.; Argiolas, A.; Torsello, A.; Locatelli, V., et al. Differen for the differential actions of ghrelin and a peptidic GHS on tial orexigenic effects of hexarelin and its analogs in the rat neural contractile response observed during binding studies hypothalamus: indication for multiple growth hormone at the motilin receptor. (Depoortere, I. Thijs, T.: Thielemans, secretagogue receptor Subtypes. Neuroendocrinology 2000, L.; Robberecht, P.; Peeters, T. L. Interaction of the growth 72, 327-332.) However, a truncated, inactive form of GHS hormone-releasing peptides ghrelin and growth hormone R1a, termed GHS-R1b, was isolated and identified during the releasing peptide-6 with the motilin receptor in the rabbit original characterization studies. Evidence is mounting that gastric antrum. J. Pharmacol. Exp. Ther: 2003, 305, 660 additional receptor subtypes could be present in different 667.) tissues to explain the diverse effects displayed by the endog 0013 The variety of activities associated with the ghrelin enous peptides and synthetic GHS. For instance, high affinity receptor could also be due to different agonists activating binding sites for ghrelin and des-acyl ghrelin have also been different signaling pathways as has been shown for ghrelin found in breast cancer cell lines, cardiomyocytes, and guinea and adenosine, both of which interact as agonists at GHS-R1a pig heart that are involved in mediating the antiproliferative, (Carreira, M.C.: Camina, J. P.; Smith, R. G.; Casanueva, F. F. cardioprotective and negative cardiac inotropic effects of Agonist-specific coupling of growth hormone secretagogue these peptides. Similarly, specific GHS binding sites besides receptor type 1a to different intracellular signaling systems. GHS-R1a and GHS-R1b have been found in prostate cancer Role of adenosine. Neuroendocrinology 2004, 79, 13-25.) cells. Further, ghrelin and des-acyl ghrelin exert different 0014. The functional activity of a GPCR has been shown effects on cell proliferation in prostate carcinoma cell lines. to often require the formation of dimers or other multimeric (Cassoni, P.; Ghé, C.; Marrocco, T.; et al. Expression of complexes with itself or other proteins. (Park, P. S.; Filipek, ghrelin and biological activity of specific receptors forghrelin S.; Wells, J.W.; Palczewski, K. Oligomerization of G protein and des-acyl ghrelin in human prostate neoplasms and related coupled receptors: past, present, and future. Biochemistry cell lines. Eur: J. Endocrinol. 2004, 150, 173-184) These 2004, 43, 15643-15656: Rios, C. D.; Jordan, B.A., Gomes, I.: various receptor Subtypes may then be implicated indepen Devi, L. A. G-protein-coupled receptor dimerization: modu dently in the wide array of biological activities displayed by lation of receptor function. Pharmacol. Ther: 2001, 92.71-87; the endogenous peptides and synthetic GHS. Indeed, Devi, L. A. Heterodimerization of G-protein-coupled recep recently, the existence of receptor Subtypes was offered as an tors: pharmacology, signaling and trafficking. Trends Phar explanation for the promotion offat accumulation by ghrelin, macol. Sci. 2001, 22,532-537.) Likewise, the activity of the despite its potent stimulation of the lipolytic hormone, growth ghrelin receptor might also be at least partially governed by hormone. (Thompson, N. M.; Gill, D. A. S.; Davies, R.; Such complexes. For example, certain reports indicate that Loveridge, N.; Houston, P. A.; Robinson, I. C. A. F.; Wells, T. interaction of GHS-R1a with GHRH(Cunha, S. R.; Mayo, K. Ghrelin and des-octanoyl ghrelin promote adipogenesis E. Ghrelin and growth hormone (GH) secreatagogues poten directly in vivo by a mechanism independent of the type 1 a tiate GH-releasing hormone (GHRH)-induced cyclic adenos growth hormone secretagogue receptor. Endocrinology 2004, ine 3',5'-monophosphate production in cells expressing trans 145, 234-242.) Further, this work suggested that the ratio of fected GHRH and GH secretagogue receptors. ghrelin and des-acyl ghrelin production could help regulate Endocrinology 2002, 143, 4570-4582: Malagon, M. M.: the balance between adipogenesis and lipolysis in response to Luque, R. M.; Ruiz-Guerrero, E.; Rodriguez-Pacheco, F.; nutritional status. Garcia-Navarro, S.; Casanueva, F. F.; Gracia-Navarro, F.; 0012. The successful creation of peptidic ghrelin ana Castafio, J. P. Intracellular signaling mechanisms mediating logues that separate the GH-modulating effects of ghrelin ghrelin-stimulated growth hormone release in Somatotropes from the effects on weight gain and appetite provides strong Endocrinology 2003, 144, 5372-5380) or between receptor evidence for the existence and physiological relevance of subtypes (Chan, C. B. Cheng, C. H. K. Identification and other receptor subtypes. (Halem, H. A.; Taylor, J. E.; Dong, J. functional characterization of two alternatively spliced Z. Shen, Y.; Datta, R.; Abizaid, A.; Diano, S.; Horvath, T.; growth hormone secretagogue receptor transcripts from the Zizzari, P.; Bluet-Pajot, M.-T.: Epelbaum, J.; Culler, M. D. pituitary of black Seabream Acanthopagrus Schlegeli. Mol. Novel analogs of ghrelin: physiological and clinical implica Cell. Endocrinol. 2004, 214, 81-95) may be involved in tions. Eur: J. Endocrinol. 2004, 151, S71-S75.) BIM-28163 modulating the function of the receptor. functions as an antagonist at the GHS-R1a receptor and inhib 0015 The vast majority of reported approaches to exploit its receptor activation by native ghrelin. However, this same ing the ghrelin receptor fortherapeutic purposes have focused molecule is a full agonist with respect to stimulating weight on modulating metabolic functions. Similarly, the vast major gain and food intake. Additionally, the existence of a still ity of literature on GHS focuses on conditions that can be uncharacterized receptor Subtype has been proposed based on treated via its GH promoting actions. Some embodiments of binding studies in various tissues that showed differences the invention described herein, in particular, take advantage between peptidic and non-peptidic GHS. (Ong, H.; Menicoll, of selective activation of the ghrelin receptor to provide an US 2011/02881 63 A1 Nov. 24, 2011
avenue for the treatment of diseases characterized by GI illeri, M. Advances in diabetic gastroparesis. Rev. Gastroen dysmotility. The improved GI motility observed with ghrelin terol. Disord. 2002, 2, 47-56; Tack et al. Gastroenterology demonstrates that ghrelin agonists may be useful in correct 2004; 126: A485; Moreaux, B.; VandenBerg, J.; Thielmans, ing conditions associated with reduced or restricted motility L.; Meulemans, A.; Coulie, B. Activation of the GHS receptor (Murray, C. D. R.; Kamm, M. A.; Bloom, S. R.; Emmanuel, accelerates gastric emptying in the dog. Digestive Disease A.V. Ghrelin for the gastroenterologist: history and potential. Week, 15-20 May 2004, New Orleans, La., USA Abstract Gastroenterology 2003, 125, 1492-1502; Fujino, K., Inui, A.; M1009; Tack et al. Gastroenterology 2004, 126: A74) Gas Asakawa, A.; Kihara, N.; Fujimura, M.; Fujimiya, M. Ghrelin troparesis (“stomach paralysis) is a syndrome characterized induces fasting motor activity of the gastrointestinal tract in by delayed gastric emptying in the absence of any mechanical conscious fed rats. J. Physiol. 2003, 550, 227-240: Edholm, obstruction. It is variably characterized by abdominal pain, T.: Levin, F.; Hellström, P. M.; Schmidt, P.T. Ghrelin stimu nausea, vomiting, weight loss, anorexia, early satiety, malnu lates motility in the Small intestine of rats through intrinsic trition, dehydration, gastroesophageal reflux, cramping and cholinergic neurons. Regul. Pept. 2004, 121, 25-30.) bloating. This chronic condition can lead to frequent hospi 0016 Included among these conditions is post-operative talization, increased disability and decreased quality of life. illeus (POI, Luckey, A.; Livingston, E.; Taché, Y. Mechanisms Severe, symptomatic gastroparesis is common in individuals and treatment of postoperative ileus. Arch. Surg. 2003, 138, suffering from diabetes, affecting from 5-10% of diabetics for 206-214; Baig, M. K. Wexner, S. D. Postoperative ileus: a a total patient population of 1 million in the U.S. alone. review. Dis. Colon Rectum 2004, 47,516-526). POI is defined Neuropathy is a frequent, debilitating complication of diabe as the impairment of GI motility that routinely occurs follow tes. Visceral neuropathy results in GI dysfunction, especially ing abdominal, intestinal, gynecological and pelvic Surgeries. involving the stomach, leading to impaired gastric motility. In the U.S. alone, 4.3 million surgeries annually induce POI, Ghrelin promotes gastric emptying both by stimulating the accounting for an economic impact of over S1 billion. POI is vagus nerve and via direct prokinetic action at the gastric considered a deleterious response to Surgical manipulation mucosa. Moreover, a recent clinical study indicates that intra with a variable duration that generally persists for 72 hours. It venous administration of the natural ghrelin peptide is an is characterized by pain, abdominal distention or bloating, effective acute therapy in diabetic gastroparesis patients. A nausea and Vomiting, accumulation of gas and fluids in the ghrelin agonist would therefore be highly effective in over bowel, and delayed passage of stool. Patients are neither able coming the fundamental motility barrierfaced by gastropare to tolerate oral feeding nor to have bowel movements until gut sis patients and correcting this condition. As with POI, no function returns. POI leads to numerous undesirable conse accepted or efficacious therapy for diabetic gastroparesis is quences, including increased patient morbidity, the costly available and most current therapies aim to provide only prolongation of hospital stays and, further, is a major cause of symptomatic relief. Further, many of the therapeutics in hospital readmission. In addition, opiate drugs given as anal development have a mechanism of action similar to earlier gesics after Surgery exacerbate this condition due to their products that have failed in this indication. Surgical proce well-recognized side effect of inhibiting bowel function. dures may ameliorate the disease process, but offer no possi Surgical manipulation of the stomach or intestine causes a bility of cure. disorganization of the gut-brain signaling pathways, impair 0018. Opioid-induced bowel dysfunction (OBD, Kurz, A.; ing GI activity and triggering POI. Ghrelin acts locally in the Sessler, D. J. Opioid-Induced Bowel Dysfunction. Drugs stomach to stimulate and coordinate the firing of vagal affer 2003, 63, 649-671.) is the term applied to the confluence of ent neurons and thereby modulate gut motility. Thus, ghrelin symptoms involving the reduced GI motility that results from accelerates gastric emptying in humans (Inui, A.; Asakawa, treatment with opioid analgesics. Approximately 40-50% of A.; Bowers, C.Y.; Mantovani, G.; Laviano, A.; Meguid, M. patients taking opioids for pain control experience OBD. It is M.; Fujimiya, M. Ghrelin, appetite, and gastric motility: the characterized by hard, dry stools, straining, incomplete emerging role of the stomach as an endocrine organ. FASEB.J. evacuation, bloating, abdominal distension and increased 2004, 18, 439-456; Peeters, T. L. Central and peripheral gastric reflux. In addition to the obvious short-term distress, mechanisms by which ghrelin regulates gut motility. J. this condition leads to physical and psychological deteriora Physiol. Pharmacol. 2003, 54(Supp. 4), 95-103.) and is a tion in patients undergoing long term opioid treatment. Fur potent agent proven to treat POI in animal models (Trudel, L.; ther, the dysfunction can be so severe as to become a dose Tomasetto, C.; Rio, M. C.; Bouin, M.; Plourde, V.; Eberling, limiting adverse effect that actually prevents adequate pain P: Poitras, P. Ghrelin/motilin-related peptide is a potent pro control. As with POI, a ghrelin agonist can be expected to kinetic to reverse gastric postoperative ileus in rats. Am. J. counteract the dysmotility resulting from opioid use. Physiol. 2002, 282, G948-G952: Trudel, L.; Bouin, M.: 0019. Two less common conditions may also be helped Tomasetto, C.; Eberling, P.; St-Pierre, S.; Bannon, P.; through the GI motility stimulation effects of ghrelin and L'Heureux, M. C.; Poitras, P. Two new peptides to improve ghrelin agonists. Short bowel syndrome is a condition that post-operative gastric ileus in dog. Peptides 2003, 24, 531 occurs after resection of a Substantial portion of Small intes 534). Ghrelin agonists duplicate the effects of ghrelin, thus tine and is characterized by malnutrition. Patients are targeting directly the underlying cause of POI to accelerate observed to have decreased ghrelin levels resulting from loss normalization of gut function and enable more rapid dis of the ghrelin-producing neuroendocrine cells of the intes charge from the hospital. Intravenous administration is often tine. It is possible the short bowel feeds back on the release of the preferred route of treatment for POI due to the impaired the hormone. (Krsek, M.; Rosicka, M.: Haluzik, M. et al. GI motility in these patients that impedes oral therapy. No Plasmaghrelin levels in patients with short bowel syndrome. agent is currently approved by the U.S. FDA specifically for Endocr: Res. 2002, 28, 27-33.) Chronic intestinal pseudo the treatment of POI. obstruction is a syndrome defined by the presence of chronic 0017. Another major motility disorder is gastroparesis, a intestinal dilation and dysmotility in the absence of mechani particular problem for both type I and type II diabetics. (Cam cal obstruction or inflammation. Both genetic and acquired US 2011/02881 63 A1 Nov. 24, 2011
causes are known to result in this disorder, which affects high (Carpino, P. Exp. Opin. Ther. Patents 2002, 12, 1599-1618.) numbers of individuals worldwide annually. (Hirano, I.; Pan Specific GHS are described in the following U.S. Pat. Nos. dolfino, J. Chronic intestinal pseudo-obstruction. Dig. Dis. and Intl. Pat. Appl. Publs. WO 89/07110; WO 89/07111; WO 2000, 18, 83-92.) 92/07578; WO93/04081; WO94/11012: WO94/13696; WO 0020. Other conditions and disorders that could be 94/19367; WO95/11029; WO95/13069; WO95/14666; WO addressed through stimulation of the ghrelin receptor are: 95/17422; WO95/17423; WO95/34311; WO 96/02530; WO emesis such as caused by cancer chemotherapy, constipation 96/15148: WO 96/22996; WO 96/22997; WO 96/24580; WO such as associated with the hypomotility phase of irritable 96/24587; WO 96/32943; WO 96/33189, WO 96/35713; WO bowel syndrome (IBS), delayed gastric emptying associated 96/38471; WO 97/00894; WO 97/06803; WO97/07117; WO with wasting conditions, gastroesophageal reflux disease 97/09060, WO 97/11697; WO 97/15191, WO 97/15573; WO (GERD), gastric ulcers (Sibilia, V.; Rindi, G.; Pagani, F.; 97/21730; WO 97/22004; WO 97/22367; WO97/22620; WO Rapetti, D.: Locatelli, V.; Torsello, A.: Campanini, N.; 97/23508; WO 97/24369; WO 97/34604: WO97/36873; WO Degenghi, R.; Netti, C. Ghrelin protects against ethanol-in 97/38709, WO 97/40023; WO 97/40071; WO 97/41878; WO duced gastric ulcers in rats: Studies on the mechanism of 97/41879; WO 97/43278; WO 97/44042; WO 97/46252; WO action. Endocrinology 2003, 144, 353–359.) and Crohn's dis 98/03473; WO 98/10653; WO 98/18815; WO 98/22124; WO CaSC. 0021 Additionally, GI dysmotility is a significant prob 98/46569, WO 98/51687; WO 98/58947; WO98/58948: WO lem in other mammals as well. For example, the motility 98/58949, WO 98/58950; WO99/08697; WO99/09991; WO dysfunction termed ileus or colic is the number one cause of 99/36431; WO99/39730; WO99/45029; WO99/58501; WO mortality among horses. Further, ileus is one of the most 99/64456; WO99/65486, WO 99/65488; WO00/01726; WO common complications of equine intestinal Surgery, in other 00/10975; WO 01/47558; WO 01/92292; WO 01/96300; WO words, post-operative ileus. This condition may also have a O1/97831; U.S. Pat. No. 3,239,345; U.S. Pat. No. 4,036,979; non-Surgical etiology. Some horses may be predisposed to U.S. Pat. No. 4411,890; U.S. Pat. No. 5,492,916; U.S. Pat. illeus based upon the anatomy and functioning of their diges No. 5,494,919; U.S. Pat. No. 5,559,128: U.S. Pat. No. 5,663, tive tract. Virtually any horse is susceptible to colic with only 171; U.S. Pat. No. 5,721,250; U.S. Pat. No. 5,721,251; U.S. minor differences based upon age, sex and breed. Addition Pat. No. 5,723,616; U.S. Pat. No. 5,726,319; U.S. Pat. No. ally, ileus may affect other animals, for example canines. 5,767,124; U.S. Pat. No. 5,798.337; U.S. Pat. No. 5,830,433; (Roussel, A.J., Jr.; Cohen, N. D.; Hooper, R. N.; Rakestraw, U.S. Pat. No. 5,919,777; U.S. Pat. No. 6,034,216: U.S. Pat. P. C. Risk factors associated with development of postopera No. 6,548,501; U.S. Pat. No. 6,559,150; U.S. Pat. No. 6,576, tive ileus in horses.J. Am. Vet. Med. Assoc. 2001, 219, 72-78; 686: U.S. Pat. No. 6,686,359; and U.S. Pat. Appl. Nos. 2002/ Van Hoogmoed, L. M.: Nieto, J. E.; Snyder, J. R.: Harmon, F. O168343; 2003/100494; 2003/130284; 2003/186844. A. Survey of prokinetic use in horses with gastrointestinal 0025 Despite this immense body of work, cyclic com injury. Vet. Surg. 2004, 33,279-285.) pounds have rarely been found to act at the receptor. When 0022 Importantly, for most of the above conditions, no they have, antagonist activity has been more prevalent. For specific, approved therapeutics exist and most therapies sim example, the 14-amino acid compound, Vapreotide, an SRIH ply address symptomatic relief. However, specific modula 14 agonistand Somatostatin mimetic, was demonstrated to be tion of the ghrelin receptor provides an opportunity to directly aghrelin antagonist. (Deghenghi R, Papotti M. Ghigo E. etal. target the site of pathophysiological disturbance to better treat Somatostatin octapeptides (lanreotide, octreotide, Vap the underlying condition and improve clinical outcome. Fur reotide, and their analogs) share the growth hormone-releas ther, unlike other agents that interact at the GHS-R1a recep ing peptide receptor in the human pituitary gland. Endocrine tor, the compounds of the invention are believed not to stimu 2001, 14, 29-33.) The binding and antagonist activities of late concurrent GH secretion. This separation of the analogues of cortistatin, a cyclic neuropeptide known to bind gastrointestinal and GH effects has not previously been nonselectively to Somatostatin receptors, to the growth hor reported for any modulators of this receptor. However, as mone secretagogue receptor have been reported (Intl. Pat. already mentioned, the existence of analogues that separate Appl. WO 03/004518). (Deghenghi R, Broglio F, Papotti M., the appetite control and GH modulatory effects associated et al. Targeting the ghrelin receptor Orally active GHS and with ghrelin has been recently reported. cortistatin analogs. Endocrine 2003, 22, 13-18) In particular, 0023 WO 01/00830 reports on short gastrointestinal pep one of these analogues, EP-01492 (cortistatin-8) has been tides (SGIP) that secrete growth hormone and also promote advanced into preclinical studies for the treatment of obesity GI motility, but these were not shown to be due to action at the as a ghrelin antagonist. These compounds exhibit an ICs of ghrelin receptor. U.S. Pat. No. 6,548,501 discloses specific 24-33 nM. In addition, these cyclic compounds and their compounds, but as GHS, useful for stimulation of GI motility. derivatives, plus their use with metal binding agents have Moreover, other endogenous factors are known to stimulate been described for their ability to be useful for radiodiagnos secretion of GH, but do not promote GI motility. Indeed, tic or radiotherapeutic use in the treatment of tumors and many actually inhibit this physiological function. Specific acromegaly. receptoragonists Such as the compounds of the present inven 0026 Cyclic and linear analogues of growth hormone tion have much better potential to be selective and effective 177-191 have been studied as treatments for obesity (WO therapeutic agents. 99/12969), with one particular compound, AOD9604, having 0024 Work has continued at the development of potent entered the clinic for this indication. A compound already and selective GHS with a number of small molecule deriva studied that is most similar to the molecules of the present tives now being known as has been recently Summarized. invention is the GHS, G-7203 (ECso-0.43 nM), the cyclic US 2011/02881 63 A1 Nov. 24, 2011
peptide analogue of the growth hormone releasing peptide, comprising an O.S or additional Natom in the ring, wherein GHRP-2 (Elias, K. A.; Ingle, G. S.; Burnier, J. P.; Hammonds, the ring is optionally substituted with Rs as defined below: G.; McDowell, R. S.; Rawson, T. E.; Somers, T. C.; Stanley, 0034 R is hydrogen or the side chain of an amino acid, or M. S.; Cronin, M. J. In vitro characterization of four novel alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem classes of growth hormone-releasing peptide. Endocrinol. bered ring, optionally comprising an O or Satom in the ring, 1995,136,5694-5699). However, simplification of this cyclic wherein the ring is optionally substituted with Rs as defined derivative led to still potent, linear compounds, whereas, for below, or alternatively RandR, or RandR togetherform compounds of the invention, linear analogues have been a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally found to be devoid of ghrelin receptor activity. comprising an O.S or additional Natom in the ring, wherein 0027. The macrocyclic compounds of the invention pos the ring is optionally substituted with Rs as defined below: sess agonist activity. As previously mentioned, however, 0035 Rs and R are each independently hydrogen or the unlike other agonists of the hCHS-R1a receptor, the com side chain of anamino acid or alternatively Rs and R together pounds of the invention unexpectedly have an insignificant form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris stimulatory effect on the release of growth hormone. Accord ing an O.S or Natom in the ring, wherein the ring is option ingly, the compounds of the present invention can exhibit ally substituted with Rs as defined below: selective action in the GI tract or for metabolic disorders 0036) R, is hydrogen, lower alkyl, substituted lower alkyl, without side effects due to GH release. cycloalkyl, Substituted cycloalkyl, a heterocyclic group, or a Substituted heterocyclic group, or alternatively R and R7 or SUMMARY OF THE INVENTION Ra and R, together form a 4-, 5-, 6-, 7- or 8-membered het 0028. The present invention provides novel conformation erocyclic ring optionally comprising an O.S or additional N ally-defined macrocyclic compounds. atom in the ring, wherein the ring is optionally Substituted 0029. These compounds can function as modulators, in with Rs as described below: particular agonists, of the ghrelin (growth hormone secreta 0037 Rs is substituted for one or more hydrogenatoms on gogue) receptor (GHS-R1a). the 3-, 4-, 5-, 6-, 7- or 8-membered ring structure and is 0030. According to aspects of the present invention, the independently selected from the group consisting of alkyl, present invention relates to compounds according to formula substituted alkyl, cycloalkyl, substituted cycloalkyl, a hetero I, II and/or III: cyclic group, a Substituted heterocyclic group, aryl, Substi tuted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, oxo, amino, halogen, formyl, acyl, carboxy, (I) carboxyalkyl, carboxyaryl, amido, carbamoyl, guanidino, R4 (CH2): 2O ureido, amidino, mercapto, Sulfonyl, Sulfonyl and Sulfona mido, or, alternatively, Rs is a fused cycloalkyl, a Substituted O sy r N R6 fused cycloalkyl, a fused heterocyclic, a substituted fused n Z heterocyclic, a fused aryl, a Substituted fused aryl, a fused heteroaryl or a substituted fused heteroaryl ring when substi (CH2) R Y. tuted for hydrogen atoms on two adjacent atoms; -Y-v \!" 0038 X is O, NR or N(R)"; R 0039 wherein R is hydrogen, lower alkyl, substituted lower alkyl, Sulfonyl, Sulfonamido oramidino and Rois hydrogen, lower alkyl, or substituted lower alkyl, or or an optical isomer, enantiomer, diastereomer, racemate or alternatively Ro and R together form a 3-, 4-, 5-, 6- or stereochemical mixture thereof, wherein: 7-membered ring, optionally comprising an O. S or 0031 R is hydrogen or the side chain of an amino acid, or additional Natom in the ring, wherein the ring is option alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem ally substituted with Rs as defined above: bered ring, optionally comprising an O. S or Natom in the 0040 Z is O or NR, ring, wherein the ring is optionally substituted with Rs as 0041 wherein R is hydrogen, lower alkyl, or substi defined below, or alternatively R and R together form a 3-, tuted lower alkyl, or alternatively R and R or R and 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S R together form a 4-, 5-, 6-, 7- or 8-membered hetero or additional Natom in the ring, wherein the ring is optionally cyclic ring, optionally comprising an O.S or additional substituted with Rs as defined below: Natom in the ring, wherein the ring is optionally Sub 0032 R is hydrogen or the side chain of an amino acid, or stituted with Rs as defined above: alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem 0042 Z is O or NR, wherein R is hydrogen, lower bered ring, optionally comprising an O. S or Natom in the alkyl, or substituted lower alkyl: ring, wherein the ring is optionally Substituted with Rs as 0043 m, n and pare each independently 0, 1 or 2: defined below; or alternatively RandR together form a 3-, 0044 T is a bivalent radical of formula IV: 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S or additional Natom in the ring, wherein the ring is optionally —U—(CH2) W Y—Z—(CH2)— (IV) substituted with Rs as defined below: 0.045 wherein dande are each independently 0, 1, 2, 3, 0033 R is hydrogen or the side chain of an amino acid, or 4 or 5: Y and Z are each optionally present; U is alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem —CRR - or —C(=O)— and is bonded to X of bered ring, optionally comprising an O or Satom in the ring, formula I; W, Y and Z are each independently selected wherein the ring is optionally substituted with Rs as defined from the group consisting of O— —NR , —S-, below, or alternatively, R and R7 or R and R togetherform SO , SO , C(=O) NH , NH C a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally (=O)—, SO. NH , NH-SO , US 2011/02881 63 A1 Nov. 24, 2011
—CRRs , —CH=CH- with the configuration Z substituted lower alkyl, except when the carbon to or E. —C=C and the ring structures below: which R- and Rs are bonded is also bonded to another heteroatom;
0050 R. R. Ras and Rs are each optionally present and, when present, are substituted for one or more hydrogen atoms on the indicated ring and each is independently selected from the group consisting of halogen, trifluoromethyl, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl, Substi tuted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car boxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, cyano, nitro, mercapto, Sulfinyl, Sulfonyl and Sulfonamido; 0051 R, is optionally present and is substituted for one or more hydrogenatoms on the indicated ring and each is independently selected from the group con sisting of alkyl, Substituted alkyl, cycloalkyl, Substi tuted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl, Substituted aryl, heteroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, amino, formyl, acyl, carboxy, carboxyalkyl, car boxyaryl, amido, carbamoyl, guanidino, ureido, ami dino, mercapto, Sulfinyl, Sulfonyl and Sulfonamido; 0052 R2s, Rao, Rao, Rs.2, Ras, R. Rae and Rs 7 are 10046 wherein G and G are each independently a each optionally present and, when no double bond is covalent bond or a bivalent radical selected from the present to the carbonatom to which it is bonded in the group consisting of —O— —NRo , —S , ring, two groups are optionally present, and when SO-, -SO. , —C(=O)—, —C(=O)—O—, present, is substituted for one hydrogen present in the —O C(=O)— —C(=O)NH , —SO. NH , ring, or when no double bond is present to the carbon —CRR - , —CH=CH- with the configuration atom to which it is bonded in the ring, is substituted Zor E, and —C=C : with G being bonded closest for one or both of the two hydrogen atoms present on to the group U, wherein any carbon atom in the rings the ring and each is independently selected from the not otherwise defined, is optionally replaced by N. group consisting of alkyl, Substituted alkyl, with the proviso that the ring cannot contain more cycloalkyl, Substituted cycloalkyl, a heterocyclic than four Natoms; K. K. K. K. and Ks are each independently O, NR or S, wherein R is as defined group, a Substituted heterocyclic group, aryl, Substi below: tuted aryl, heteroaryl, substituted heteroaryl, 0047 R and R are each independently hydrogen, hydroxy, alkoxy, aryloxy, oxo, amino, formyl, acyl, lower alkyl, or substituted lower alkyl, or alternatively carboxy, carboxyalkyl, carboxyaryl, amido, carbam RandR togetherform a 3- to 12-membered cyclic oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, ring optionally comprising one or more heteroatoms sulfonyl, sulfonamido and, only if a double bond is selected from the group consisting of O, S and N, present to the carbon atom to which it is bonded, wherein the ring is optionally substituted with Rs as halogen; and defined above; 0053 Rao and Ra are each independently hydrogen, 0048 R2, Ro and Ra are each independently lower alkyl, substituted lower alkyl, R as defined hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substi above, or alternatively Rao and R together form a 3 tuted cycloalkyl, heterocyclic, substituted heterocy to 12-membered cyclic ring optionally comprising clic, aryl, substituted aryl, heteroaryl, substituted het one or more heteroatoms selected from the group eroaryl, formyl, acyl, carboxyalkyl, carboxyaryl, consisting of O. Sand N wherein the ring is optionally amido, amidino, Sulfonyl or Sulfonamido; substituted with Rs as defined above, or alternatively 0049 R2 and Rs are each independently hydrogen, one of Rao and R is hydroxy, alkoxy, aryloxy, amino, lower alkyl, substituted lower alkyl, R, wherein mercapto, carbamoyl, amidino, ureido or guanidino, R is a side chain of an amino acid Such as a standard or unusual amino acid, or alternatively Ra and Rs while the other is hydrogen, lower alkyl or substituted togetherform a 3- to 12-membered cyclic ring option lower alkyl, except when the carbon to which Rao and ally comprising one or more heteroatoms selected Rare bonded is also bonded to another heteroatom; from the group consisting of O, S and N, or alterna- 0.054 with the proviso that T is not an amino acid resi tively one of R or Rs is hydroxy, alkoxy, aryloxy, due, dipeptide fragment, tripeptide fragment or higher amino, mercapto, carbamoyl, amidino, ureido or order peptide fragment comprising standard amino guanidino while the other is hydrogen, lower alkyl or acids; US 2011/02881 63 A1 Nov. 24, 2011
-continued (II) !'l- R69 G1 !'l- R69 G G2 G2
0064 wherein G and G are as defined above, and wherein any carbon atom in the ring is optionally replaced by N, with the proviso that the aromatic ring cannot contain more than four N atoms and the cycloalkyl ring cannot contain more than two N or an optical isomer, enantiomer, diastereomer, racemate or atoms; stereochemical mixture thereof, wherein: 0065 R is hydrogen, alkyl, substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic 0055 Rso is —(CH2)CH, —CH(CH)(CH2)CH, group, a Substituted heterocyclic group, aryl, Substi —(CH2)CH(CH), —C(CH), —(CHRss), Rs, or tuted aryl, heteroaryl, substituted heteroaryl, formyl, —CH(ORs)CH, whereinss is 1, 2 or 3; tt is 1 or 2: uu is 0. acyl, carboxyalkyl, carboxyaryl, amido, amidino, Sul 1 or 2; and VV is 0, 1, 2, 3 or 4: Riss is hydrogen or C-C alkyl: fonyl or sulfonamido: Rs is amino, hydroxy, alkoxy, cycloalkyl or Substituted 0066 R and Rea are each independently hydrogen, cycloalkyl, and Rs7 is hydrogen, alkyl, acyl, amino acyl, lower alkyl, substituted lower alkyl or R.; or alter Sulfonyl, carboxyalkyl or carboxyaryl; natively Res and R together form a 3- to 12-mem 0056 Rs is hydrogen, C-C alkyl or C-C alkyl substi bered cyclic ring optionally comprising one or more tuted with hydroxy or alkoxy; heteroatoms selected from the group consisting of O, 0057 Rs is —(CHRs).Rs, wherein ww is 0, 1, 2 or 3: S and N, or alternatively one of R and R is Rss is hydrogen, C-C alkyl, amino, hydroxy or alkoxy. Rs hydroxy, alkoxy, aryloxy, amino, mercapto, carbam is aryl, substituted aryl, heteroaryl, substituted heteroaryl, oyl, amidino, ureido or guanidino, while the other is cycloalkyl or substituted cycloalkyl: hydrogen, lower alkyl or substituted lower alkyl, 0058 Rs is hydrogen or C-C alkyl: except when the carbon to which R and R are 0059 X is O, NR or N(R): bonded is also bonded to anotherheteroatom; and R. 0060 wherein R is hydrogen, lower alkyl, substituted indicates the side chain of an amino acid such as a lower alkyl, Sulfonyl, Sulfonamido oramidino and Rois standard or unusual amino acid; hydrogen, lower alkyl, or substituted lower alkyl: 0067. Res and Res are each optionally present, and, when present are substituted for one or more hydro 0061 Zs is O or NR, wherein R is hydrogen, lower genatoms on the ring and each is independently halo alkyl, or substituted lower alkyl; and gen, trifluoromethyl, alkyl, Substituted alkyl, 0062 T is a bivalent radical of formula V: cycloalkyl, Substituted cycloalkyl, a heterocyclic U -(CH2) W. Y. Z-(CH2). (V) group, a Substituted heterocyclic group, aryl, Substi tuted aryl, heteroaryl, substituted heteroaryl, 0063 wherein d and e are independently 0, 1, 2, 3, 4 or hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car 5, Y, and Z are each optionally present; U is boxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, —CROR— or —C(=O)— and is bonded to X of guanidino, ureido, amidino, cyano, nitro, mercapto, formula II, wherein Reo and R are each independently sulfinyl, sulfonyl or sulfonamido: hydrogen, lower alkyl, or substituted lower alkyl, or 0068 Roe and Re, are each optionally present, and alternatively R and R together form a 3- to 12-mem when no double bond is present to the carbon atom to bered cyclic ring optionally comprising one or more which it is bonded in the ring, two groups are option heteroatoms selected from the group consisting of O.S ally present, and, when present, each is Substituted for and N, wherein the ring is optionally substituted with Rs. one hydrogen present in the ring, or when no double as defined above: W.Y. and Z are each independently bond is present to the carbon atom to which it is Selected from the group consisting of —O—, bonded in the ring, is substituted for one or both of the two hydrogen atoms present on the ring and each is independently alkyl, Substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted het with the configuration Z or E. —C=C- and the ring erocyclic, aryl, Substituted aryl, heteroaryl, Substi structures depicted below: tuted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, amino, formyl, acyl, carboxy, carboxyalkyl, car
boxyaryl, amido, carbamoyl, guanidino, ureido, ami dino, mercapto, Sulfinyl, Sulfonyl, Sulfonamido and, 1sy G1 only if a double bond is present to the carbon atom to G t e O which it is bonded, halogen; 0069 R is optionally present, and when present is 2. R, Substituted for one or more hydrogen atoms on the ring and each is independently alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocy US 2011/02881 63 A1 Nov. 24, 2011
clic group, a Substituted heterocyclic group, aryl, Sub stituted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, Oxo, amino, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, carbam oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, sulfonyl or sulfonamido; (0070 K is O or S; and (0071 ff is 1, 2, 3, 4 or 5: 0072 with the proviso that T is not an amino acid residue, dipeptide fragment, tripeptide fragment or higher order peptide fragment comprising standard amino acids; J 0073 or -" and
R (III) 73 O R72 ( O NH HN R75 '. R71 N T R F4 S x:1 nz, O Rio 0080 wherein E. E. E. E. and Es are each optionally present and when present are each independently Selected from the group consisting of halogen, trifluo or an optical isomer, enantiomer, diastereomer, racemate or romethyl, alkyl, substituted alkyl, cycloalkyl, substi stereochemical mixture thereof, wherein: tuted cycloalkyl, a heterocyclic group, a substituted het 007.4 Rio is hydrogen, C-C alkyl or alternatively Rio erocyclic group, aryl, Substituted aryl, heteroaryl, and R, together form a 4-, 5-, 6-, 7- or 8-membered ring, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, cyano, optionally comprising an O. Nor Satom in the ring, wherein Sulfinyl, Sulfonyl and Sulfonamido, and represent Sub the ring is optionally substituted with Rs, as defined below: stitution at one or more available positions on the mono 0075 R is hydrogen, —(CH2)CH, —CH(CH)(CH) cyclic or bicyclic aromatic ring, wherein said Substitu CH, -(CH2)CH(CH), —(CH) R-7 or —CH tion is made with the same or different selected group (OR77)CH or, alternatively R7 and Ryo together form a 4-, member, and J and J are each independently O or S; 5-, 6-, 7- or 8-membered ring, optionally comprising an O. N 0081 Rs. Rs, and Rs are each independently substituted or Satom in the ring, wherein the ring is optionally Substi for one or more hydrogen atoms on the 3-, 4-, 5-, 6-, 7- or tuted with Rs, as defined below: wherein aa is 0, 1, 2, 3, 4 or 8-membered ring structure and are independently selected 5: bb is 1, 2 or 3; cc is 0, 1, 2 or 3; and dd is 0,1,2,3 or 4: R. from the group consisting of alkyl, Substituted alkyl, is aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a cycloalkyl or substituted cycloalkyl, R77 is hydrogen, alkyl, Substituted heterocyclic group, aryl, Substituted aryl, het acyl, aminoacyl, Sulfonyl, carboxyalkyl or carboxyaryl; eroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, oXo, amino, halogen, formyl, acyl, carboxy, carboxyalkyl, 0076 R, is C-C alkyl; or alternatively R7 and R, carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally mercapto, Sulfinyl, Sulfonyl and Sulfonamido, or, alterna comprising an O or S atom in the ring, wherein the ring is tively, Rs. Rs, and Rs are each independently a fused optionally substituted with Rs, as defined below: cycloalkyl, a Substituted fused cycloalkyl, a fused heterocy 0077 R, is hydrogen, or alternatively R7 and R, clic, a Substituted fused heterocyclic, a fused aryl, a Substi together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally tuted fused aryl, a fused heteroaryl or a substituted fused comprising an O.S or Natom in the ring, wherein the ring is heteroaryl ring when Substituted for hydrogen atoms on two optionally substituted with Rs, as defined below: adjacent atoms; 0078 Riza is hydrogen or C-C alkyl or alternatively R. I0082 X is O, NRs or N(R)"; and R7s together form a 3-, 4-, 5-, 6- or 7-membered ring, 0.083 wherein R is hydrogen, lower alkyl, substituted optionally comprising an O. Nor Satom in the ring, wherein lower alkyl, sulfonyl, Sulfonamido oramidino and Rois the ring is optionally substituted with Rs as defined below: hydrogen, lower alkyl, or substituted lower alkyl: 0079 R7s is —(CHR 7s)R or alternatively R-7s and R. I0084 Zo is O or NR, wherein R is hydrogen, lower together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally alkyl, or substituted lower alkyl; and comprising an O, N or Satom in the ring, wherein the ring is I0085 T is the same as defined for T with the exception optionally substituted with Rs as defined below; wherein R-7s that U is bonded to X of formula III. is hydrogen, C-C alkyl, amino, hydroxy or alkoxy, and Rio I0086 According to further aspects of the present inven is selected from the group consisting of the following struc tion, the compound is a ghrelin receptor agonist or a GHS tures: R1a receptor agonist. US 2011/02881 63 A1 Nov. 24, 2011
0087 Further aspects of the present invention provide 0099 FIG. 5 (panels A through E) shows concentration pharmaceutical compositions comprising: (a) a compound of response curves for activation of the hCHS-R1a receptor by the present invention; and (b) a pharmaceutically acceptable exemplary compounds of the present invention. carrier, excipient or diluent. 0100 FIG. 6 shows graphs depicting pharmacokinetic 0088 Additional aspects of the present invention provide parameters for exemplary compounds of the present inven kits comprising one or more containers containing pharma tion, specifically after oral administration of 8 mg/kg com ceutical dosage units comprising an effective amount of one pound 298 (panel A), after Subcutaneous injection of 2 mg/kg or more compounds of the present invention packaged with compound 298 with cyclodextrin (panel B), after intravenous optional instructions for the use thereof. administration of 2 mg/kg compound 25 with cyclodextrin 0089 Aspects of the present invention further provide (panel C) and after intravenous administration of 2 mg/kg methods of stimulating gastrointestinal motility, modulating compound 298 with cyclodextrin (panel D). GHS-R1a receptor activity in a mammal and/or treating a 0101 FIG. 7 (panels A and B) shows graphs presenting gastrointestinal disorder comprising administering to a Sub effects on gastric emptying for exemplary compounds of the ject in need thereof an effective amount of a modulator that present invention. modulates a mammalian GHS-R1a receptor. In particular 0102 FIG. 8 shows a graph presenting effects on postop embodiments, interaction of the modulator and the GHS-R1a erative ileus for an exemplary compound of the present inven receptor does not result in a significant amount of growth tion. hormone release. In still other embodiments, the modulator is 0103 FIG.9 (panels A through D) shows graphs depicting a compound of formula I, II and/or III. the effect on pulsatile growth hormone release for an exem 0090. Additional aspects of the present invention provide plary compound of the present invention. methods of diagnosing tumors and/or acromegaly, compris 0104 FIG. 10 shows a competitive binding curve for bind ing administering compounds of the present invention and a ing of an exemplary compound of the present invention to the radiolabeled metal binding agent and detecting the binding of hCHS-R1a receptor. the composition to a biological target, and treating tumors 0105 FIG. 11 shows an activation curve demonstrating and/or acromegaly comprising administering a therapeuti the agonism of an exemplary compound of the present inven cally effective amount of a composition comprising a com tion. pound of the present invention. 0106 FIG. 12 shows agraph depicting the lack of effect on 0091. Further aspects of the present invention relate to ghrelin-induced growth hormone release for an exemplary methods of making the compounds of formula I, II and/or III. compound of the present invention. 0092 Aspects of the present invention further relate to 0107 FIG. 13 shows graphs depicting receptor desensiti methods of preventing and/or treating disorders described Zation associated with binding of exemplary compounds of herein, in particular, gastrointestinal disorders, including the present invention to the hCHS-R1a receptor. post-operative ileus, gastroparesis, such as diabetic and post 0.108 FIG. 14 (panels A and B) shows graphs presenting Surgical gastroparesis, opioid-induced bowel dysfunction, effects on gastric emptying for an exemplary compound of chronic intestinal pseudo-obstruction, short bowel syndrome, the present invention. emesis such as caused by cancer chemotherapy, constipation 0109 FIG. 15 shows a graph presenting effects on post such as associated with the hypomotility phase of irritable operative ileus for an exemplary compound of the present bowel syndrome (IBS), delayed gastric emptying associated invention. with wasting conditions, gastroesophageal reflux disease 0110 FIG. 16 shows graphs depicting reversal of mor (GERD), gastric ulcers, Crohn's disease, gastrointestinal dis phine-delayed gastric emptying (panel A) and morphine-de orders characterized by dysmotility and other diseases and layed gastrointestinal transit (panel B) for an exemplary com disorders of the gastrointestinal tract. pound of the present invention. 0093. The present invention also relates to compounds of 0111 FIG. 17 (panels A and B) shows graphs depicting formula I, II and/or III used for the preparation of a medica effects on gastroparesis for exemplary compounds of the ment for prevention and/or treatment of the disorders described herein. present invention. 0094. The foregoing and other aspects of the present invention are explained in greater detail in the specification DETAILED DESCRIPTION set forth below. 0112 The foregoing and other aspects of the present invention will now be described in more detail with respect to BRIEF DESCRIPTION OF THE DRAWINGS other embodiments described herein. It should be appreciated that the invention can be embodied in different forms and 0095 FIG. 1 shows a scheme presenting a general syn should not be construed as limited to the embodiments set thetic strategy to provide conformationally-defined macro forth herein. Rather, these embodiments are provided so that cycles of the present invention. this disclosure will be thorough and complete, and will fully 0096 FIG.2 shows a general thioester strategy for making convey the scope of the invention to those skilled in the art. macrocyclic compounds of the present invention. 0113. The terminology used in the description of the 0097 FIG. 3 shows a general ring-closing metathesis invention herein is for the purpose of describing particular (RCM) strategy for macrocyclic compounds of the present embodiments only and is not intended to be limiting of the invention. invention. As used in the description of the invention and the 0098 FIG. 4 (panels A through E) shows competitive appended claims, the singular forms “a”, “an and “the are binding curves for binding of exemplary compounds of the intended to include the plural forms as well, unless the con present invention to the hGHS-R1a receptor. text clearly indicates otherwise. Additionally, as used herein, US 2011/02881 63 A1 Nov. 24, 2011
the term “and/or includes any and all combinations of one or number of heteroatoms in each ring is four or less and each more of the associated listed items and may be abbreviated as ring contains at least one carbon atom. The fused rings com f'. pleting the bicyclic or tricyclic heterocyclic groups may con 0114. Unless otherwise defined, all technical and scien tain only carbon atoms and may be saturated or partially tific terms used herein have the same meaning as commonly unsaturated. The N and Satoms may optionally be oxidized understood by one of ordinary skill in the art to which this and the Natoms may optionally be quaternized. Heterocyclic invention belongs. also refers to alkyl groups containing said monocyclic, bicy 0115 All publications, U.S. patent applications, U.S. pat clic or tricyclic heterocyclic groups. Examples of heterocy ents and other references cited herein are incorporated by clic rings include, but are not limited to, 2- or 3-piperidinyl, 2 reference in their entireties. or 3-piperazinyl, 2- or 3-morpholinyl. All such heterocyclic 0116. The term “alkyl refers to straight or branched chain groups may also be optionally Substituted as described below saturated or partially unsaturated hydrocarbon groups having 0.122 The term "heteroaryl” refers to an aromatic group in from 1 to 20 carbon atoms, in some instances 1 to 8 carbon a single or fused ring system having from 5 to 15 ring atoms, atoms. The term “lower alkyl refers to alkyl groups contain in some instances 5 to 10, which have at least one heteroatom ing 1 to 6 carbonatoms. Examples of alkyl groups include, but in at least one of the rings, said heteroatom being selected are not limited to, methyl, ethyl, isopropyl, tert-butyl, 3-hex from O, S or N. Each ring of the heteroaryl group can contain enyl, and 2-butynyl. By “unsaturated' is meant the presence one or two Oatoms, one or two Satoms, one to four Natoms, of 1, 2 or 3 double or triple bonds, or a combination of the two. provided that the total number of heteroatoms in each ring is Such alkyl groups may also be optionally Substituted as four or less and each ring contains at least one carbon atom. described below. The fused rings completing the bicyclic or tricyclic groups 0117. When a subscript is used with reference to an alkyl may contain only carbon atoms and may be saturated, par or other hydrocarbon group defined herein, the subscript tially unsaturated or aromatic. In structures where the lone refers to the number of carbon atoms that the group may pair of electrons of a nitrogen atom is not involved in com contain. For example, C-C alkyl indicates an alkyl group pleting the aromatic pi electron system, the N atoms may with 2, 3 or 4 carbon atoms. optionally be quaternized or oxidized to the N-oxide. Het 0118. The term “cycloalkyl refers to saturated or partially eroaryl also refers to alkyl groups containing said cyclic unsaturated cyclic hydrocarbon groups having from 3 to 15 groups. Examples of monocyclic heteroaryl groups include, carbonatoms in the ring, in Some instances 3 to 7, and to alkyl but are not limited to pyrrolyl pyrazolyl pyrazolinyl, imida groups containing said cyclic hydrocarbon groups. Examples Zolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiaz of cycloalkyl groups include, but are not limited to, cyclopro olyl, furanyl, thienyl, oxadiazolyl pyridyl, pyrazinyl, pyrim pyl, cyclopropylmethyl, cyclopentyl, 2-(cyclohexyl)ethyl, idinyl, pyridazinyl, and triazinyl. Examples of bicyclic cycloheptyl, and cyclohexenyl. Cycloalkyl as defined herein heteroaryl groups include, but are not limited to indolyl, also includes groups with multiple carbon rings, each of benzothiazolyl, benzoxazolyl, benzothienyl, quinolinyl, tet which may be saturated or partially unsaturated, for example rahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, ben decalinyl, 2.2.1-bicycloheptanyl or adamantanyl. All Such Zopyranyl, indolizinyl, benzopyranyl, isobenzofuranyl. cycloalkyl groups may also be optionally substituted as chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinox described below. alinyl, indazolyl, purinyl, pyrrolopyridinyl, furopyridinyl, 0119 The term “aromatic” refers to an unsaturated cyclic thienopyridinyl, dihydroisoindolyl, and tetrahydroquinoli hydrocarbon group having a conjugated pi electron system nyl. Examples of tricyclic heteroaryl groups include, but are that contains 4n+2 electrons where n is an integer greater than not limited to carbazolyl, benzindolyl, phenanthrollinyl, or equal to 1. Aromatic molecules are typically stable and are acridinyl, phenanthridinyl, and Xanthenyl. All Such heteroaryl depicted as a planar ring of atoms with resonance structures groups may also be optionally substituted as described below. that consist of alternating double and single bonds, for (0123. The term “hydroxy” refers to the group –OH. example benzene or naphthalene. 0.124. The term “alkoxy' refers to the group —OR, 0120. The term “aryl refers to an aromatic group in a wherein R is alkyl, cycloalkyl or heterocyclic. Examples single or fused carbocyclic ring system having from 6 to 15 include, but are not limited to methoxy, ethoxy, tert-butoxy, ring atoms, in some instances 6 to 10, and to alkyl groups cyclohexyloxy and tetrahydropyranyloxy. containing said aromatic groups. Examples of aryl groups 0.125. The term “aryloxy' refers to the group —OR, include, but are not limited to, phenyl, 1-naphthyl 2-naphthyl wherein R, is aryl or heteroaryl. Examples include, but are not and benzyl. Aryl as defined herein also includes groups with limited to phenoxy, benzyloxy and 2-maphthyloxy. multiple aryl rings which may be fused, as in naphthyl and 0.126 The term “acyl refers to the group —C(=O)—R. anthracenyl, or unfused, as in biphenyl and terphenyl. Aryl wherein R is alkyl, cycloalkyl, heterocyclic, aryl or het also refers to bicyclic or tricyclic carbon rings, where one of eroaryl. Examples include, but are not limited to, acetyl, the rings is aromatic and the others of which may be saturated, benzoyl and furoyl. partially unsaturated or aromatic, for example, indanyl or I0127. The term “amino acyl indicates an acyl group that tetrahydronaphthyl (tetralinyl). All Such aryl groups may also is derived from an amino acid. be optionally substituted as described below. I0128. The term “amino” refers to an —NRR group 0121. The term “heterocycle” or "heterocyclic” refers to wherein R and Rare independently selected from the group saturated or partially unsaturated monocyclic, bicyclic or tri consisting of hydrogen, alkyl, cycloalkyl, heterocyclic, aryl cyclic groups having from 3 to 15 atoms, in Some instances 3 and heteroaryl. Alternatively, R and R together form a het to 7, with at least one heteroatom in at least one of the rings, erocyclic ring of 3 to 8 members, optionally substituted with said heteroatom being selected from O, S or N. Each ring of unsubstituted alkyl, unsubstituted cycloalkyl, unsubstituted the heterocyclic group can contain one or two Oatoms, one or heterocyclic, unsubstituted aryl, unsubstituted heteroaryl, two S atoms, one to four Natoms, provided that the total hydroxy, alkoxy, aryloxy, acyl, amino, amido, carboxy, car US 2011/02881 63 A1 Nov. 24, 2011 boxyalkyl, carboxyaryl, mercapto, Sulfinyl, Sulfonyl, Sul cycloalkyl, heterocyclic, aryl or heteroaryl. Alternatively, R, fonamido, amidino, carbamoyl, guanidino or ureido, and and R together form a heterocyclic ring of 3 to 8 members, optionally containing one to three additional heteroatoms optionally substituted with unsubstituted alkyl, unsubstituted selected from O, S or N. cycloalkyl, unsubstituted heterocyclic, unsubstituted aryl, 0129. The term "amido” refers to the group —C(=O)— unsubstituted heteroaryl, hydroxy, alkoxy, aryloxy, acyl, NRR, wherein RandR are independently selected from the amino, amido, carboxy, carboxyalkyl, carboxyaryl, mer group consisting of hydrogen, alkyl, cycloalkyl, heterocyclic, capto, Sulfinyl, Sulfonyl, Sulfonamido, amidino, carbamoyl, aryland heteroaryl. Alternatively, R, and R together form a guanidino or ureido, and optionally containing one to three heterocyclic ring of 3 to 8 members, optionally substituted additional heteroatoms selected from O, S or N. with unsubstituted alkyl, unsubstituted cycloalkyl, unsubsti 0145 The term “carbamoyl refers to a group of the for tuted heterocyclic, unsubstituted aryl, unsubstituted het mula - N(R)—C(=O) OR, wherein R, is selected from eroaryl, hydroxy, alkoxy, aryloxy, acyl, amino, amido, car hydrogen, alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl; boxy, carboxyalkyl, carboxyaryl, mercapto, Sulfinyl, and R is selected from alkyl, cycloalkyl, heterocylic, aryl or Sulfonyl, Sulfonamido, amidino, carbamoyl, guanidino or heteroaryl. ureido, and optionally containing one to three additional het 0146 The term “guanidino” refers to a group of the for eroatoms selected from O, S or N. mula N(R) C(=NR) NR,R, wherein R. R. and R, 0130. The term "amidino” refers to the group—C(=NR) are independently selected from hydrogen, alkyl, cycloalkyl, NR,R, wherein R, is selected from the group consisting of heterocyclic, aryl or heteroaryl. Alternatively, R, and R, hydrogen, alkyl, cycloalkyl, heterocyclic, aryland heteroaryl; together form a heterocyclic ring or 3 to 8 members, option and R, and R, are independently selected from the group ally substituted with unsubstituted alkyl, unsubstituted consisting of hydrogen, alkyl, cycloalkyl, heterocyclic, aryl cycloalkyl, unsubstituted heterocyclic, unsubstituted aryl, and heteroaryl. Alternatively, R, and R, together form a het unsubstituted heteroaryl, hydroxy, alkoxy, aryloxy, acyl, erocyclic ring of 3 to 8 members, optionally substituted with amino, amido, carboxy, carboxyalkyl, carboxyaryl, mer unsubstituted alkyl, unsubstituted cycloalkyl, unsubstituted capto, Sulfinyl, Sulfonyl, Sulfonamido, amidino, carbamoyl, heterocyclic, unsubstituted aryl, unsubstituted heteroaryl, guanidino or ureido, and optionally containing one to three hydroxy, alkoxy, aryloxy, acyl, amino, amido, carboxy, car additional heteroatoms selected from O, S or N. boxyalkyl, carboxyaryl, mercapto, Sulfinyl, Sulfonyl, Sul 0147 The term “ureido” refers to a group of the formula fonamido, amidino, carbamoyl, guanidino or ureido, and —N(R)—C(=O)—NRR, wherein R, R and R, are optionally containing one to three additional heteroatoms independently selected from hydrogen, alkyl, cycloalkyl, het selected from O, S or N. erocyclic, aryl or heteroaryl. Alternatively, R, and R. 0131 The term “carboxy” refers to the group -COH. together form a heterocyclic ring of 3 to 8 members, option 0132) The term “carboxyalkyl refers to the group ally substituted with unsubstituted alkyl, unsubstituted —COR, wherein R is alkyl, cycloalkyl or heterocyclic. cycloalkyl, unsubstituted heterocyclic, unsubstituted aryl, 0133. The term “carboxyaryl” refers to the group unsubstituted heteroaryl, hydroxy, alkoxy, aryloxy, acyl, —COR, wherein R, is aryl or heteroaryl. amino, amido, carboxy, carboxyalkyl, carboxyaryl, mer 0134. The term "cyano” refers to the group —CN. capto, Sulfinyl, Sulfonyl, Sulfonamido, amidino, carbamoyl, 0135. The term “formyl refers to the group –C(=O)H, guanidino or ureido, and optionally containing one to three also denoted —CHO. additional heteroatoms selected from O, S or N. 0136. The term “halo,” “halogen” or “halide” refers to 0.148. The term “optionally substituted' is intended to fluoro, fluorine or fluoride, chloro, chlorine or chloride, expressly indicate that the specified group is unsubstituted or bromo, bromine or bromide, and iodo, iodine or iodide, substituted by one or more suitable substituents, unless the respectively. optional Substituents are expressly specified, in which case 0137 The term “oxo” refers to the bivalent group —O, the term indicates that the group is unsubstituted or Substi which is Substituted in place of two hydrogen atoms on the tuted with the specified substituents. As defined above, vari same carbon to form a carbonyl group. ous groups may be unsubstituted or Substituted (i.e., they are 0.138. The term “mercapto' refers to the group —SR, optionally substituted) unless indicated otherwise herein wherein R is hydrogen, alkyl, cycloalkyl, heterocyclic, aryl (e.g., by indicating that the specified group is unsubstituted). or heteroaryl. 0149. The term “substituted when used with the terms 0.139. The term “nitro” refers to the group - NO. alkyl, cycloalkyl, heterocyclic, aryland heteroaryl refers to an 0140. The term “trifluoromethyl” refers to the group alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl group hav —CF. ing one or more of the hydrogen atoms of the group replaced I014.1) The term "sulfinyl" refers to the group S(=O)R. by substituents independently selected from unsubstituted wherein R is alkyl, cycloalkyl, heterocyclic, aryl or het alkyl, unsubstituted cycloalkyl, unsubstituted heterocyclic, eroaryl. unsubstituted awl, unsubstituted heteroaryl, hydroxy, alkoxy, 0142. The term "sulfonyl refers to the group —S(=O) aryloxy, acyl, amino, amido, carboxy, carboxyalkyl, car 2 R, wherein R is alkyl, cycloalkyl, heterocyclic, aryl or boxyaryl, halo, oxo, mercapto, Sulfinyl, Sulfonyl, Sulfona heteroaryl. mido, amidino, carbamoyl, guanidino, ureido and groups of 0143. The term “aminosulfonyl refers to the group the formulas —NRC(=O)R - NRC(=NR)R. —NR2 S(=O). Rs wherein R2 is hydrogen, alkyl, OC(=O)NR,R, OC(=O)R —OC(=O)CR cycloalkyl, heterocyclic, aryl or heteroaryl; and R is alkyl, —NRSO.R., or NRSONRN, wherein R. R. cycloalkyl, heterocyclic, aryl or heteroaryl. Ree Rt. R. R. R. R. R. R. R., and R, are indepen 0144. The term “sulfonamido” refers to the group dently selected from hydrogen, unsubstituted alkyl, unsubsti —S(=O) NR,R, wherein R, and R are independently tuted cycloalkyl, unsubstituted heterocyclic, unsubstituted selected from the group consisting of hydrogen, alkyl, aryl or unsubstituted heteroaryl; and wherein RandR are US 2011/02881 63 A1 Nov. 24, 2011 independently selected from unsubstituted alkyl, unsubsti 0157. The term “agonist” refers to a compound that dupli tuted cycloalkyl, unsubstituted heterocyclic, unsubstituted cates at least some of the effect of the endogenous ligand of a aryl or unsubstituted heteroaryl. Alternatively, R and R. protein, receptor, enzyme or the like. RandR, or RandR, together form a heterocyclic ring of 0158. The term “antagonist” refers to a compound that 3 to 8 members, optionally substituted with unsubstituted inhibits at least some of the effect of the endogenous ligand of alkyl, unsubstituted cycloalkyl, unsubstituted heterocyclic, a protein, receptor, enzyme or the like. unsubstituted aryl, unsubstituted heteroaryl, hydroxy, alkoxy, 0159. The term “growth hormone secretagogue' (GHS) aryloxy, acyl, amino, amido, carboxy, carboxyalkyl, car refers to any exogenously administered compound or agent boxyaryl, mercapto, Sulfinyl, Sulfonyl, Sulfonamido, ami that directly or indirectly stimulates or increases the endog dino, carbamoyl, guanidino orureido, and optionally contain enous release of growth hormone, growth hormone-releasing ing one to three additional heteroatoms selected from O, S or hormone, or somatostatin in an animal, in particular, a human. N. In addition, the term “substituted for awl and heteroaryl A GHS may be peptidic or non-peptidic in nature, in some groups includes as an option having one of the hydrogen instances, with an agent that can be administered orally. In atoms of the group replaced by cyano, nitro or trifluorom Some instances, the agent can induce a pulsatile response. ethyl. 0160 The term “modulator” refers to a compound that 0150. A substitution is made provided that any atom's imparts an effect on a biological or chemical process or normal valency is not exceeded and that the Substitution mechanism. For example, a modulator may increase, facili results in a stable compound. Generally, when a Substituted tate, upregulate, activate, inhibit, decrease, block, prevent, form of a group is present, Such Substituted group is prefer delay, desensitize, deactivate, down regulate, or the like, a ably not further substituted or, if substituted, the substituent biological or chemical process or mechanism. Accordingly, a comprises only a limited number of Substituted groups, in modulator can be an "agonist oran'antagonist.” Exemplary Some instances 1, 2, 3 or 4 such substituents. biological processes or mechanisms affected by a modulator 0151. When any variable occurs more than one time in any include, but are not limited to, receptor binding and hormone constituent or in any formula herein, its definition on each release or secretion. Exemplary chemical processes or occurrence is independent of its definition at every other mechanisms affected by a modulator include, but are not occurrence. Also, combinations of Substituents and/or vari limited to, catalysis and hydrolysis. ables are permissible only if such combinations result in 0.161 The term “variant' when applied to a receptor is stable compounds. meant to include dimers, trimers, tetramers, pentamers and 0152. A “stable compound” or “stable structure” refers to other biological complexes containing multiple components. a compound that is Sufficiently robust to Survive isolation to a These components can be the same or different. useful degree of purity and formulation into an efficacious 0162 The term "peptide' refers to a chemical compound therapeutic agent. comprised of two or more amino acids covalently bonded 0153. The term “amino acid” refers to the common natural together. (genetically encoded) or synthetic amino acids and common 0163 The term "peptidomimetic' refers to a chemical derivatives thereof, known to those skilled in the art. When compound designed to mimic a peptide, but which contains applied to amino acids, “standard” or “proteinogenic’ refers structural differences through the addition or replacement of to the genetically encoded 20 amino acids in their natural one of more functional groups of the peptide in order to configuration. Similarly, when applied to amino acids, modulate its activity or other properties, such as solubility, "unnatural or “unusual refers to the wide selection of non metabolic stability, oral bioavailability, lipophilicity, perme natural, rare or synthetic amino acids Such as those described ability, etc. This can include replacement of the peptide bond, by Hunt, S. in Chemistry and Biochemistry of the Amino side chain modifications, truncations, additions of functional Acids, Barrett, G. C., Ed., Chapman and Hall: New York, groups, etc. When the chemical structure is not derived from 1985. the peptide, but mimics its activity, it is often referred to as a 0154 The term “residue” with reference to an amino acid “non-peptide peptidomimetic.” or amino acid derivative refers to a group of the formula: 0164. The term "peptide bond refers to the amide - C (=O)—NH functionality with which individual amino acids are typically covalently bonded to each other in a pep H tide. N (CH2) 0.165. The term “protecting group' refers to any chemical compound that may be used to prevent a potentially reactive R. O functional group. Such as an amine, a hydroxyl or a carboxyl, on a molecule from undergoing a chemical reaction while chemical change occurs elsewhere in the molecule. A number wherein R is an amino acid side chain, and n=0, 1 or 2 in of such protecting groups are known to those skilled in the art this instance. and examples can be found in “Protective Groups in Organic 0155 The term “fragment with respect to a dipeptide, Synthesis.” Theodora W. Greene and Peter G. Wuts, editors, tripeptide or higher order peptide derivative indicates a group John Wiley & Sons, New York, 3' edition, 1999 [ISBN that contains two, three or more, respectively, amino acid 0471160199. Examples of amino protecting groups include, residues. but are not limited to, phthalimido, trichloroacetyl, benzy 0156 The term "amino acid side chain” refers to any side loxycarbonyl, tert-butoxycarbonyl, and adamantyloxycarbo chain from a standard or unnatural amino acid, and is denoted nyl. In some embodiments, amino protecting groups are car R. For example, the side chain of alanine is methyl, the side bamate amino protecting groups, which are defined as an chain of valine is isopropyl and the side chain of tryptophanis amino protecting group that when bound to an amino group 3-indolylmethyl. forms a carbamate. In other embodiments, amino carbamate US 2011/02881 63 A1 Nov. 24, 2011 protecting groups are allyloxycarbonyl (Alloc), benzyloxy uct on the polymer precipitated through the addition of carbonyl (Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), tert diethyl ether and processed as a solid. This has been termed butoxycarbonyl (Boc) and O.C.-dimethyl-3,5-dimethoxyben “liquid-phase' chemistry. Zyloxycarbonyl (Ddz). For a recent discussion of newer nitrogen protecting groups: Theodoridis, G. Tetrahedron (0170 The term “linker when used in reference to solid 2000, 56, 2339-2358. Examples of hydroxyl protecting phase chemistry refers to a chemical group that is bonded groups include, but are not limited to, acetyl, tert-butyldim covalently to a solid support and is attached between the ethylsilyl (TBDMS), trityl (Trt), tert-butyl, and tetrahydropy Support and the Substrate typically in order to permit the ranyl (THP). Examples of carboxyl protecting groups release (cleavage) of the Substrate from the Solid Support. include, but are not limited to methyl ester, tert-butyl ester, However, it can also be used to impart stability to the bond to benzyl ester, trimethylsilylethyl ester, and 2.2.2-trichloroet the Solid Support or merely as a spacer element. Many solid hyl ester. Supports are available commercially with linkers already 0166 The term "solid phase chemistry” refers to the con attached. duct of chemical reactions where one component of the reac 0171 Abbreviations used for amino acids and designation tion is covalently bonded to a polymeric material (solid Sup of peptides follow the rules of the IUPAC-IUB Commission port as defined below). Reaction methods for performing of Biochemical Nomenclature in J. Biol. Chem. 1972, 247, chemistry on Solid phase have become more widely known 977-983. This document has been updated: Biochem. J., and established outside the traditional fields of peptide and 1984, 219,345-373 Eur: J. Biochem., 1984, 138, 9-37; 1985, oligonucleotide chemistry. 152, 1 Internat. J. Pept. Prot. Res., 1984, 24, following p 84. (0167. The term “solid support,”99 “solid&g phase” or “resin' J. Biol. Chem., 1985,260, 14-42; Pure Appl. Chem., 1984,56, refers to a mechanically and chemically stable polymeric 595-624. Amino Acids and Peptides, 1985, 16,387-410; and matrix utilized to conduct solid phase chemistry. This is in Biochemical Nomenclature and Related Documents, 2nd denoted by “Resin,” “P or the following symbol: edition, Portland Press, 1992, pp. 39-67. Extensions to the rules were published in the JCBN/NC-IUB Newsletter 1985, 1986, 1989; see Biochemical Nomenclature and Related Documents, 2nd edition, Portland Press, 1992, pp 68-69. 0172. The term “effective amount or “effective' is 0168 Examples of appropriate polymer materials include, intended to designate a dose that causes a relief of symptoms but are not limited to, polystyrene, polyethylene, polyethyl of a disease or disorder as noted through clinical testing and ene glycol, polyethylene glycol grafted or covalently bonded evaluation, patient observation, and/or the like, and/or a dose to polystyrene (also termed PEG-polystyrene, TentaGelTM, that causes a detectable change in biological or chemical Rapp, W.; Zhang, L.; Bayer, E. In Innovations and Perspec activity. The detectable changes may be detected and/or fur tives in Solid Phase Synthesis. Peptides, Polypeptides and ther quantified by one skilled in the art for the relevant mecha Oligonucleotides; Epton, R., Ed., SPCC Ltd.: Birmingham, nism or process. AS is generally understood in the art, the UK; p 205), polyacrylate (CLEARTM), polyacrylamide, poly dosage will vary depending on the administration routes, urethane, PEGA polyethyleneglycol poly(N,N-dimethy symptoms and body weight of the patient but also depending lacrylamide) co-polymer, Meldal, M. Tetrahedron Lett. 1992, upon the compound being administered. 33,3077-3080, cellulose, etc. These materials can optionally 0173 Administration of two or more compounds “in com contain additional chemical agents to form cross-linked bination” means that the two compounds are administered bonds to mechanically stabilize the structure, for example closely enough in time that the presence of one alters the polystyrene cross-linked with divinylbenezene (DVB, usu biological effects of the other. The two compounds can be ally 0.1-5%, preferably 0.5-2%). This solid support can administered simultaneously (concurrently) or sequentially. include as non-limiting examples aminomethyl polystyrene, Simultaneous administration can be carried out by mixing the hydroxymethyl polystyrene, benzhydrylamine polystyrene compounds prior to administration, or by administering the (BHA), methylbenzhydrylamine (MBHA) polystyrene, and other polymeric backbones containing free chemical func compounds at the same point in time but at differentanatomic tional groups, most typically, —NH or —OH, for further sites or using different routes of administration. The phrases derivatization or reaction. The term is also meant to include “concurrent administration', 'administration in combina “Ultraresins” with a high proportion (“loading) of these tion', 'simultaneous administration' or “administered simul functional groups such as those prepared from polyethylene taneously as used herein, means that the compounds are imines and cross-linking molecules (Barth, M.; Rademann, J. administered at the same point in time or immediately fol J. Comb. Chem. 2004, 6,340-349). At the conclusion of the lowing one another. In the latter case, the two compounds are synthesis, resins are typically discarded, although they have administered at times sufficiently close that the results been shown to be able to be reused such as in Frechet, J. M. J.; observed are indistinguishable from those achieved when the Haque, K. E. Tetrahedron Lett. 1975, 16, 3055. compounds are administered at the same point in time. 0169. In general, the materials used as resins are insoluble 0.174. The term “pharmaceutically active metabolite' is polymers, but certain polymers have differential solubility intended to mean a pharmacologically active product pro depending on Solvent and can also be employed for Solid duced through metabolism in the body of a specified com phase chemistry. For example, polyethylene glycol can be pound. utilized in this manner since it is soluble in many organic 0.175. The term “solvate” is intended to mean a pharma Solvents in which chemical reactions can be conducted, but it ceutically acceptable Solvate form of a specified compound is insoluble in others, such as diethyl ether. Hence, reactions that retains the biological effectiveness of Such compound. can be conducted homogeneously in Solution, then the prod Examples of solvates, without limitation, include compounds US 2011/02881 63 A1 Nov. 24, 2011
of the invention in combination with water, isopropanol, etha nol, methanol, DMSO, ethyl acetate, acetic acid, or ethano (X) -copyed lamine. n U 2 1. Compounds S 0176 Novel macrocyclic compounds of the present inven O tion include macrocyclic compounds comprising a building block structure including a tether component that undergoes (Z2) cyclization to form the macrocyclic compound. The building y block structure can comprise amino acids (standard and U O unnatural), hydroxy acids, hydrazino acids, aza-amino acids, specialized moieties such as those that play a role in the introduction of peptide Surrogates and isosteres, and a tether component as described herein. The tether component can be y selected from the following: U (Z2), \-O Jaaa
(0177 wherein (Z) is the site of a covalent bond of T to Z, and Z is as defined below for formula I, and wherein (X) is the site of a covalent bond of T to X, and X is as defined below for formula I; L, is —CH2— or —O—; U is —CRoRo or —C(=O)—, Roo is lower alkyl; Ro and Ro are each independently hydrogen, lower alkyl or substituted lower alkyl; XX is 2 or 3; yy is 1 or 2: ZZ is 1 or 2; and aaa is 0 or 1. 0.178 Macrocyclic compounds of the present invention further include those of formula I, formula II and/or formula (X) III: L2 U1
kr(Z) (I) U N(x) N (Z)
(CH2) O (CH2) - x y 2 p R r-4,
or an optical isomer, enantiomer, diastereomer, racemate or stereochemical mixture thereof, 0179 wherein: 0180 R is hydrogen or the side chain of an amino acid, or alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem bered ring, optionally comprising an O. S or N atom in the ring, wherein the ring is optionally Substituted with Rs as defined below, or alternatively R and R together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S or additional Natom in the ring, wherein the ring is optionally substituted with Rs as defined below: 0181 R is hydrogen or the side chain of an amino acid, or alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem bered ring, optionally comprising an O. S or N atom in the ring, wherein the ring is optionally substituted with Rs as defined below; or alternatively RandR together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S or additional Natom in the ring, wherein the ring is optionally substituted with Rs as defined below: US 2011/02881 63 A1 Nov. 24, 2011
0182 R is hydrogen or the side chain of an amino acid, or 0194 wherein dande are each independently 0, 1, 2, 3, alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem 4 or 5: Y and Z are each optionally present; U is bered ring, optionally comprising an O or Satom in the ring, —CRR - or —C(=O)— and is bonded to X of wherein the ring is optionally substituted with Rs as defined formula I; W, Y and Z are each independently selected below, or alternatively, R and R7 or R and R togetherform from the group consisting of O— —NR , —S-, a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally SO , SO. , —O C(=O)—, —C(=O) comprising an O.S or additional Natom in the ring, wherein NH-, -NH C(=O)-, -SO. NH-, -NH the ring is optionally substituted with Rs as defined below: SO. , —CRRs , —CH=CH- with the configu 0183 R is hydrogen or the side chain of an amino acid, or ration Zor E. —C=C- and the ring structures below: alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem bered ring, optionally comprising an O or Satom in the ring, wherein the ring is optionally substituted with Rs as defined below, or alternatively RandR, or RandR togetherform a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally comprising an O.S or additional Natom in the ring, wherein the ring is optionally substituted with Rs as defined below: 0184 Rs and R are each independently hydrogen or the side chain of anamino acid or alternatively Rs and R together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris ing an O.S or Natom in the ring, wherein the ring is option ally substituted with Rs as defined below: 0185. R, is hydrogen, lower alkyl, substituted lower alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, or a Substituted heterocyclic group, or alternatively R and R7 or Ra and R, together form a 4-, 5-, 6-, 7- or 8-membered het erocyclic ring optionally comprising an O.S or additional N atom in the ring, wherein the ring is optionally Substituted with Rs as described below: 0186 Rs is substituted for one or more hydrogenatoms on the 3-, 4-, 5-, 6-, 7- or 8-membered ring structure and is independently selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, a hetero cyclic group, a Substituted heterocyclic group, aryl, Substi tuted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, Oxo, amino, halogen, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, guanidino, (0195 wherein G and G are each independently a ureido, amidino, mercapto, Sulfinyl, Sulfonyl and Sulfona covalent bond or a bivalent radical selected from the mido, or, alternatively, Rs is a fused cycloalkyl, a Substituted group consisting of —O— —NRo , —S—, fused cycloalkyl, a fused heterocyclic, a substituted fused SO s SO, s C(=O) s C(=) O s heterocyclic, a fused aryl, a Substituted fused aryl, a fused —C(=O)NH , NH-C(=O) , SO, heteroaryl or a substituted fused heteroaryl ring when substi NH , NH-SO , CRoR . tuted for hydrogen atoms on two adjacent atoms; —CH=CH with the configuration Z or E, and —C=C-, with G being bonded closest to the group 0187 X is O, NR or N(R): U; wherein any carbonatom in the rings not otherwise 0188 wherein R is hydrogen, lower alkyl, substituted defined, is optionally replaced by N, with the proviso lower alkyl, Sulfonyl, Sulfonamido oramidino and Rois that the ring cannot contain more than four Natoms; hydrogen, lower alkyl, or substituted lower alkyl, or K. K. K. K. and Ks are each independently O, alternatively Ro and R together form a 3-, 4-, 5-, 6- or NR or S, wherein R is as defined below: 7-membered ring, optionally comprising an O. S or 0.196 R2 and R are each independently hydrogen, additional Natom in the ring, wherein the ring is option lower alkyl, or substituted lower alkyl, or alternatively ally substituted with Rs as defined above: RandR togetherform a 3- to 12-membered cyclic (0189 Z is O or NR, ring optionally comprising one or more heteroatoms (0190 wherein R is hydrogen, lower alkyl, or substi selected from the group consisting of O, S and N, tuted lower alkyl, or alternatively R and R or R and wherein the ring is optionally substituted with Rs as R together form a 4-, 5-, 6-, 7- or 8-membered hetero defined above; cyclic ring, optionally comprising an O.S or additional 0.197 R2, Ro and Ra are each independently Natom in the ring, wherein the ring is optionally Sub hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substi stituted with Rs as defined above; tuted cycloalkyl, heterocyclic, substituted heterocy (0191 Z is O or NR, wherein R is hydrogen, lower clic, aryl, substituted aryl, heteroaryl, substituted het alkyl, or substituted lower alkyl: eroaryl, formyl, acyl, carboxyalkyl, carboxyaryl, amido, amidino, Sulfonyl or Sulfonamido; 0.192 m, n and pare each independently 0, 1 or 2: 0198 R2 and Rs are each independently hydrogen, 0193 T is a bivalent radical of formula IV: lower alkyl, substituted lower alkyl, R, wherein R is a side chain of an amino acid Such as a standard US 2011/02881 63 A1 Nov. 24, 2011 17
or unusual amino acid, or alternatively Ra and Rs 0203 with the proviso that T is not an amino acid resi togetherform a 3- to 12-membered cyclic ring option due, dipeptide fragment, tripeptide fragment or higher ally comprising one or more heteroatoms selected order peptide fragment comprising standard amino from the group consisting of O, S and N, or alterna acids; tively one of R or Rs is hydroxy, alkoxy, aryloxy, amino, mercapto, carbamoyl, amidino, ureido or guanidino while the other is hydrogen, lower alkyl or substituted lower alkyl, except when the carbon to which R- and Rs are bonded is also bonded to (II) another heteroatom; H O 0199 R. R. Rs and Rs are each optionally present and, when present, are Substituted for one or Rs. ( more hydrogen atoms on the indicated ring and each O N HN H is independently selected from the group consisting of Y.53 av R R50 T2 52 halogen, trifluoromethyl, alkyl, Substituted alkyl, S 14 N S X Z O cycloalkyl, Substituted cycloalkyl, a heterocyclic H 2 5 group, a Substituted heterocyclic group, aryl, Substi tuted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car boxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, cyano, nitro, mercapto, or an optical isomer, enantiomer, diastereomer, racemate or Sulfinyl, Sulfonyl and Sulfonamido; stereochemical mixture thereof, wherein: 0200 R, is optionally present and is substituted for 0204 Rs is —(CH), CH, —CH(CH)(CH), CH, one or more hydrogenatoms on the indicated ring and —(CH2)CH(CH), —C(CH), —(CHRss), Rs, or each is independently selected from the group con sisting of alkyl, Substituted alkyl, cycloalkyl, Substi —CH(ORs)CH, whereinss is 1, 2 or 3; tt is 1 or 2: uu is 0. tuted cycloalkyl, a heterocyclic group, a Substituted 1 or 2; and VV is 0, 1, 2, 3 or 4: Riss is hydrogen or C-C alkyl: heterocyclic group, aryl. Substituted aryl, heteroaryl, Rs is amino, hydroxy, alkoxy, cycloalkyl or Substituted Substituted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, cycloalkyl, and Rs7 is hydrogen, alkyl, acyl, amino acyl, amino, formyl, acyl, carboxy, carboxyalkyl, car Sulfonyl, carboxyalkyl or carboxyaryl; boxyaryl, amido, carbamoyl, guanidino, ureido, ami 0205 Rs is hydrogen, C-C alkyl or C-C alkyl substi dino, mercapto, Sulfinyl, Sulfonyl and Sulfonamido; tuted with hydroxy or alkoxy; 0201 R2s, Rao, Rao, Rs.2, Ras, R34, Rao and Rs 7 are 0206 Rs is —(CHRss)Rs, wherein ww is 0, 1, 2 or 3: each optionally present and, when no double bond is Rss is hydrogen, C-C alkyl, amino, hydroxy or alkoxy; Rso present to the carbonatom to which it is bonded in the is aryl, substituted aryl, heteroaryl, substituted heteroaryl, ring, two groups are optionally present, and when cycloalkyl or substituted cycloalkyl: present, is Substituted for one hydrogen present in the 0207 Rs is hydrogen or C-C alkyl; ring, or when no double bond is present to the carbon atom to which it is bonded in the ring, is substituted (0208 X is O, NRs or N(R)"; for one or both of the two hydrogen atoms present on 0209 wherein R is hydrogen, lower alkyl, substituted the ring and each is independently selected from the lower alkyl, sulfonyl, Sulfonamido oramidino and Rois group consisting of alkyl, Substituted alkyl, hydrogen, lower alkyl, or substituted lower alkyl: cycloalkyl, Substituted cycloalkyl, a heterocyclic 0210 Zs is O or NR, wherein R is hydrogen, lower group, a Substituted heterocyclic group, aryl, Substi alkyl, or substituted lower alkyl; and tuted aryl, heteroaryl, substituted heteroaryl, 0211 T, is a bivalent radical of formula V: hydroxy, alkoxy, aryloxy, Oxo, amino, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, carbam U—(CH2) W. Y. Z -(CH2). (V) oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, 0212 wherein d and e are independently 0, 1, 2, 3, 4 or sulfonyl, sulfonamido and, only if a double bond is 5: Y, and Z are each optionally present; U is present to the carbon atom to which it is bonded, —CROR— or —C(=O)— and is bonded to X of halogen; and formula II, wherein Reo and Rare each independently 0202 Rao and Ra are each independently hydrogen, hydrogen, lower alkyl, or substituted lower alkyl, or lower alkyl, substituted lower alkyl, R as defined alternatively R and R together form a 3- to 12-mem above, or alternatively Rao and R together form a 3 bered cyclic ring optionally comprising one or more to 12-membered cyclic ring optionally comprising heteroatoms selected from the group consisting of O.S one or more heteroatoms selected from the group and N, wherein the ring is optionally substituted with Rs. consisting of O, S and N wherein the ring is optionally as defined above: W.Y. and Z are each independently substituted with Rs as defined above, or alternatively Selected from the group consisting of —O—, one of Rao and R is hydroxy, alkoxy, aryloxy, amino, mercapto, carbamoyl, amidino, ureido or guanidino, while the other is hydrogen, lower alkyl or substituted lower alkyl, except when the carbon to which Rao and —CH=CH with the configuration Z or E. Rare bonded is also bonded to another heteroatom; —C=C , and the ring structures depicted below: US 2011/02881 63 A1 Nov. 24, 2011 18
only if a double bond is present to the carbon atom to
which it is bonded, halogen; R65 0218 R is optionally present, and when present is 1sé G1 Substituted for one or more hydrogen atoms on the Y ring and each is independently alkyl, Substituted G "f alkyl, cycloalkyl, Substituted cycloalkyl, a heterocy 2. R. clic group, a Substituted heterocyclic group, aryl, Sub stituted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, oxo, amino, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, carbam oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, sulfonyl or sulfonamido; 0219) K is O or S; and 0213 wherein G and G are as defined above, and wherein any carbon atom in the ring is optionally 0220 ff is 1, 2, 3, 4 or 5: replaced by N, with the proviso that the aromatic ring 0221) with the proviso that T is not an amino acid cannot contain more than four N atoms and the residue, dipeptide fragment, tripeptide fragment or cycloalkyl ring cannot contain more than two N higher order peptide fragment comprising standard atoms; amino acids; or 0214 R is hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl, Substi (III) tuted aryl, heteroaryl, substituted heteroaryl, formyl, R73 O acyl, carboxyalkyl, carboxyaryl, amido, amidino, Sul * - / fonyl or sulfonamido: O NH HN R75 0215 Re and Rea are each independently hydrogen, lower alkyl, substituted lower alkyl or R. or alter N T R F4 natively Res and R together form a 3- to 12-mem S x1 ''nz, O bered cyclic ring optionally comprising one or more Rio heteroatoms selected from the group consisting of O, S and N, or alternatively one of R and R is hydroxy, alkoxy, aryloxy, amino, mercapto, carbam or an optical isomer, enantiomer, diastereomer, racemate or oyl, amidino, ureido or guanidino, while the other is stereochemical mixture thereof, wherein: hydrogen, lower alkyl or substituted lower alkyl, 0222 Rao is hydrogen, C-C alkyl or alternatively Rio except when the carbon to which Re and Rea are and R, together form a 4-, 5-, 6-, 7- or 8-membered ring, bonded is also bonded to another heteroatom; and R. optionally comprising an O. Nor Satom in the ring, wherein indicates the side chain of a standard or unusual the ring is optionally substituted with Rs, as defined below: amino acid; 0223 R-7 is hydrogen, —(CH2)CH, —CH(CH)(CH) 0216 Res and Rs are each optionally present, and, CH, -(CH2)CH(CH), —(CH) R-7 or —CH when present are substituted for one or more hydro (OR77)CH or, alternatively R, and Rio together form a 4-, genatoms on the ring and each is independently halo 5-, 6-, 7- or 8-membered ring, optionally comprising an O. N gen, trifluoromethyl, alkyl, Substituted alkyl, or S atom in the ring, wherein the ring is optionally Substi cycloalkyl, Substituted cycloalkyl, a heterocyclic tuted with Rs, as defined below: wherein aa is 0, 1, 2, 3, 4 or group, a Substituted heterocyclic group, aryl, Substi 5: bb is 1, 2 or 3; cc is 0, 1, 2 or 3; and dd is 0, 1, 2, 3 or 4: Rze tuted aryl, heteroaryl, substituted heteroaryl, is aryl, substituted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car cycloalkyl or Substituted cycloalkyl, R77 is hydrogen, alkyl, boxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, acyl, aminoacyl, Sulfonyl, carboxyalkyl or carboxyaryl; guanidino, ureido, amidino, cyano, nitro, mercapto, 0224 R is C-C alkyl; or alternatively R7 and R7 sulfinyl, sulfonyl or sulfonamido: together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally 0217 R and Re, are each optionally present, and comprising an O or S atom in the ring, wherein the ring is when no double bond is present to the carbon atom to which it is bonded in the ring, two groups are option optionally substituted with Rs, as defined below: ally present, and, when present, each is Substituted for 0225 R7 is hydrogen, or alternatively R7 and R7 one hydrogen present in the ring, or when no double together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally bond is present to the carbon atom to which it is comprising an O.S or Natom in the ring, wherein the ring is bonded in the ring, is substituted for one or both of the optionally substituted with Rs, as defined below: two hydrogen atoms present on the ring and each is 0226 R is hydrogen or C-C alkyl or alternatively R. independently alkyl, Substituted alkyl, cycloalkyl, and R7s together form a 3-, 4-, 5-, 6- or 7-membered ring, substituted cycloalkyl, heterocyclic, substituted het optionally comprising an O. Nor Satom in the ring, wherein erocyclic, aryl, Substituted aryl, heteroaryl, Substi the ring is optionally substituted with Rs as defined below: tuted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, 0227 R-7s is —(CHR-7s)Ro or alternatively R7s and R amino, formyl, acyl, carboxy, carboxyalkyl, car together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally boxyaryl, amido, carbamoyl, guanidino, ureido, ami comprising an O, N or Satom in the ring, wherein the ring is dino, mercapto, Sulfinyl, Sulfonyl, Sulfonamido and, optionally substituted with Rs as defined below; wherein R-7s US 2011/02881 63 A1 Nov. 24, 2011 19 is hydrogen, C-C alkyl, amino, hydroxy or alkoxy, and Rio 0234 In some embodiments of the present invention, the is selected from the group consisting of the following struc compound can have one of the following structures: tures:
E4 -- / and
0228 wherein E. E. E. E. and Es are each optionally present and when present are each independently Selected from the group consisting of halogen, trifluo romethyl, alkyl, substituted alkyl, cycloalkyl, substi tuted cycloalkyl, a heterocyclic group, a Substituted het erocyclic group, aryl, Substituted aryl, heteroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, cyano, Sulfinyl, Sulfonyl and Sulfonamido, and represent Sub stitution at one or more available positions on the mono cyclic or bicyclic aromatic ring, wherein said Substitu tion is made with the same or different selected group member, and J and J are each independently O or S; 0229 Rs. Rs, and Rs are each independently substituted for one or more hydrogen atoms on the 3-, 4-, 5-, 6-, 7- or 8-membered ring structure and are independently selected from the group consisting of alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl, Substituted aryl, het eroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, oxo, amino, halogen, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, mercapto, Sulfinyl, Sulfonyl and Sulfonamido, or, alterna tively, Rs. Rs, and Rs are each independently a fused cycloalkyl, a Substituted fused cycloalkyl, a fused heterocy clic, a Substituted fused heterocyclic, a fused aryl, a Substi tuted fused aryl, a fused heteroaryl or a substituted fused heteroaryl ring when Substituted for hydrogen atoms on two adjacent atoms; 0230 X is O, NRs or N(R)"; 0231 wherein R is hydrogen, lower alkyl, substituted lower alkyl, sulfonyl, sulfonamido oramidino and Rois hydrogen, lower alkyl, or substituted lower alkyl: 0232 Zo is O or NR, wherein R is hydrogen, lower alkyl, or substituted lower alkyl; and 0233 T is the same as defined for T with the exception that U is bonded to X of formula III. US 2011/02881 63 A1 Nov. 24, 2011 20
-continued -continued US 2011/02881 63 A1 Nov. 24, 2011 21
-continued -continued US 2011/02881 63 A1 Nov. 24, 2011 22
-continued -continued 2. O
N N O \ )- NH HN
C
C
O
O
Cl,
or an optical isomer, enantiomer, diastereomer, racemate or stereochemical mixture thereof. 0235. The present invention includes isolated compounds. An isolated compound refers to a compound that, in some embodiments, comprises at least 10%, at least 25%, at least US 2011/02881 63 A1 Nov. 24, 2011
50% or at least 70% of the compounds of a mixture. In some 0241. Likewise, many geometric isomers of double bonds embodiments, the compound, pharmaceutically acceptable and the like can also be present in the compounds disclosed salt thereof or pharmaceutical composition containing the herein, and all such stable isomers are included within the compound exhibits a statistically significant binding and/or present invention unless otherwise specified. Also included in antagonist activity when tested in biological assays at the the invention are tautomers and rotamers of formula I, II human ghrelin receptor. and/or III. 0236. In the case of compounds, salts, or solvates that are 0242. The use of the following symbols at the right refers solids, it is understood by those skilled in the art that the to Substitution of one or more hydrogenatoms of the indicated inventive compounds, salts, and Solvates may exist in differ ring with the defined substituent R. ent crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas. R 0237. The compounds of formula I, II and/or III disclosed SA (O, S, NH) herein have asymmetric centers. The inventive compounds may exist as single stereoisomers, racemates, and/or mixtures C.21 C.MYR ofenantiomers and/or diastereomers. All Such single stereoi Somers, racemates, and mixtures thereof are intended to be within the scope of the present invention. In particular 0243 The use of the following symbol indicates a single embodiments, however, the inventive compounds are used in bond or an optional double bond: ----. optically pure form. The terms “S” and “R” configuration as 0244 Embodiments of the present invention further pro used herein are as defined by the IUPAC 1974 Recommen vide intermediate compounds formed through the synthetic dations for Section E, Fundamentals of Stereochemistry methods described herein to provide the compounds of for (Pure Appl. Chem. 1976, 45, 13-30.) mula I, II and/or III. The intermediate compounds may pos 0238. Unless otherwise depicted to be a specific orienta sess utility as a therapeutic agent for the range of indications tion, the present invention accounts for all Stereoisomeric described herein and/or a reagent for further synthesis meth forms. The compounds may be prepared as a single stereoi ods and reactions. Somer or a mixture of stereoisomers. The non-racemic forms may be obtained by either synthesis or resolution. The com 2. Synthetic Methods pounds may, for example, be resolved into the component 0245. The compounds of formula I, II and/or II can be enantiomers by standard techniques, for example formation synthesized using traditional Solution synthesis techniques or of diastereomeric pairs via salt formation. The compounds Solid phase chemistry methods. In either, the construction also may be resolved by covalently bonding to a chiral moi involves four phases: first, synthesis of the building blocks ety. The diastereomers can then be resolved by chromato comprising recognition elements for the biological target graphic separation and/or crystallographic separation. In the receptor, plus one tether moiety, primarily for control and case of a chiral auxiliary moiety, it can then be removed. As an definition of conformation. These building blocks are alternative, the compounds can be resolved through the use of assembled together, typically in a sequential fashion, in a chiral chromatography. Enzymatic methods of resolution second phase employing standard chemical transformations. could also be used in certain cases. The precursors from the assembly are then cyclized in the 0239. As generally understood by those skilled in the art, third stage to provide the macrocyclic structures. Finally, the an “optically pure' compound is one that contains only a post-cyclization processing fourth stage involving removal of single enantiomer. As used herein, the term “optically active' protecting groups and optional purification provides the is intended to mean a compound comprising at least a suffi desired final compounds. Synthetic methods for this general cient excess of one enantiomer over the other such that the type of macrocyclic structure are described in Intl. Pat. Appls. mixture rotates plane polarized light. Optically active com WO 01/25257, WO 2004/111077, WO 2005/012331 and WO pounds have the ability to rotate the plane of polarized light. 2005/012332, including purification procedures described in The excess of one enantiomer over another is typically WO 2004/111077 and WO 2005/O12331. expressed as enantiomeric excess (e.e.). In describing an opti 0246. In some embodiments of the present invention, the cally active compound, the prefixes D and L or R and S are macrocyclic compounds of formula I, II and/or III may be used to denote the absolute configuration of the molecule synthesized using Solid phase chemistry on a soluble or about its chiral center(s). The prefixes “d' and “1” or (+) and insoluble polymer matrix as previously defined. For solid (-) are used to denote the optical rotation of the compound phase chemistry, a preliminary stage involving the attachment (i.e., the direction in which a plane of polarized light is rotated of the first building block, also termed “loading to the resin by the optically active compound). The “1” or (-) prefix must be performed. The resin utilized for the present inven indicates that the compound is levorotatory (i.e., rotates the tion preferentially has attached to it a linker moiety, L. These plane of polarized light to the left or counterclockwise) while linkers are attached to an appropriate free chemical function the “d' or (+) prefix means that the compound is dextrarota ality, usually an alcohol or amine, although others are also tory (i.e., rotates the plane of polarized light to the right or possible, on the base resin through standard reaction methods clockwise). The sign of optical rotation, (-) and (+), is not known in the art, such as any of the large number of reaction related to the absolute configuration of the molecule, Rand S. conditions developed for the formation of ester or amide 0240. A compound of the invention having the desired bonds. Some linker moieties for the present invention are pharmacological properties will be optically active and, can designed to allow for simultaneous cleavage from the resin be comprised of at least 90% (80% e.e.), at least 95% (90% with formation of the macrocycle in a process generally e.e.), at least 97.5% (95% e.e.) or at least 99% (98% e.e.) of a termed “cyclization-release.” (van Maarseveen, J. H. Solid single isomer. phase synthesis of heterocycles by cyclization/cleavage US 2011/02881 63 A1 Nov. 24, 2011 24 methodologies. Comb. Chem. High Throughput Screen. of manufacturing the compounds of the present invention 1998, 1, 185-214; Ian W. James, Linkers for solid phase comprising (a) assembling building block structures, (b) organic synthesis. Tetrahedron 1999, 55, 4855-4946; Eggen chemically transforming the building block structures, (c) weiler, H.-M. Linkers for solid-phase synthesis of small mol cyclizing the building block structures including a tether ecules: coupling and cleavage techniques. Drug Discovery component, (d) removing protecting groups from the building Today 1998, 3, 552-560; Backes, B. J.; Ellman, J. A. Solid block structures, and (e) optionally purifying the product support linker strategies. Curr. Opin. Chem. Biol. 1997. 1, obtained from step (d). In some embodiments, assembly of 86-93. Of particular utility in this regard for compounds of the the building block structures may be sequential. In further invention is the 3-thiopropionic acid linker. (Hojo, H.; embodiments, the synthesis methods are carried out using Aimoto, S. Bull. Chem. Soc. Jpn. 1991, 64. 111-117; Zhang, traditional solution synthesis techniques or solid phase chem L.; Tam, J.J. Am. Chem. Soc. 1999, 121,3311-3320.) istry techniques. 0247 Such a process provides material of higher purity as 0251 A. Amino Acids only cyclic products are released from the Solid Support and 0252 Amino acids, Boc- and Fmoc-protected amino acids minimal contamination with the linear precursor occurs as and side chain protected derivatives, including those of would happen in Solution phase. After sequential assembly of N-methyl and unnatural amino acids, were obtained from all the building blocks and tether into the linear precursor commercial Suppliers for example Advanced ChemTech using known or standard reaction chemistry, base-mediated (Louisville, Ky., USA), Bachem (Bubendorf, Switzerland), intramolecular attack on the carbonyl attached to this linker ChemImpex (Wood Dale, Ill., USA), Novabiochem (subsid by an appropriate nucleophilic functionality that is part of the iary of Merck KGaA, Darmstadt, Germany), PepTech (Bur tether building block results information of the amide orester lington, Mass., USA), Synthetech (Albany, Oreg., USA) or bond that completes the cyclic structure as shown (Scheme 1). synthesized through standard methodologies known to those An analogous methodology adapted to Solution phase can in the art. Ddz-amino acids were either obtained commer also be applied as would likely be preferable for larger scale cially from Orpegen (Heidelberg, Germany) or Advanced applications. ChemTech (Louisville, Ky., USA) or synthesized using stan
Scheme 1. Cwclization-release Stratve O O ls
Y BB-BB-BB Base Cyclization-release Linker BB-BB-BB Hip (Y= O, NH)
HY- ) Tether
0248 Although this description accurately represents the dard methods utilizing Ddz-OPhor Ddz-N. (Birr, C.: Loch pathway for one of the methods of the present invention, the inger, W.; Stahnke, G.; Lang, P. The O.C.-dimethyl-3,5- thioester Strategy, another method of the present invention, dimethoxybenzyloxycarbonyl (Ddz) residue, al that of ring-closing metathesis (RCM), proceeds through a N-protecting group labile toward weak acids and irradiation. modified route where the tether component is actually Justus Liebigs Ann. Chem. 1972, 763, 162-172.) Bts-amino assembled during the cyclization step. However, in the RCM methodology as well, assembly of the building blocks pro acids were synthesized by known methods. (Vedes, E.; Lin, ceeds sequentially, followed by cyclization (and release from S.: Klapara, A.; Wang, J. “Heteroarene-2-sulfonyl Chlorides the resin if solid phase). An additional post-cyclization pro (BtsC1, ThsCl): Reagents for Nitrogen Protection and >99% cessing step is required to remove particular byproducts of the Racemization-Free Phenylglycine Activation with SOCl. 'J. RCM reaction, but the remaining Subsequent processing is Am. Chem. Soc. 1996, 118,9796-9797. Also WO 01/25257, done in the same manner as for the thioester or analogous WO 2004/111077)N-Alkylamino acids, in particular N-me base-mediated cyclization strategy. thyl amino acids, are commercially available from multiple 0249 Moreover, it will be understood that steps including vendors (Bachem, Novabiochem, Advanced ChemTech, the methods provided herein may be performed indepen ChemImpex). In addition, N-alkyl amino acid derivatives dently or at least two steps may be combined. Additionally, were accessed via literature methods. (Hansen, D. W., Jr.; steps including the methods provided herein, when per Pilipauskas, D.J. Org. Chem. 1985, 50,945-950.) formed independently or combined, may be performed at the same temperature or at different temperatures without depart 0253 B. Tethers ing from the teachings of the present invention. 0254 Tethers were obtained from the methods previously 0250 Novel macrocyclic compounds of the present inven described in Intl. Pat. Appl. WO 01/25257, WO 2004/111077, tion include those formed by a novel process including WO 2005/012331 and U.S. Provisional Patent Application cyclization of a building block structure to form a macrocy Ser. No. 60/622,055. Procedures for synthesis of tethers as clic compound comprising a tether component described described herein are presented in the Examples below. Exem herein. Accordingly, the present invention provides methods plary tethers (T) include, but are not limited to, the following: US 2011/02881 63 A1 Nov. 24, 2011 25
-continued y (Z) U O
O y U (Z), \- O )aaa
and intermediates in the manufacture thereof, wherein (Z) is the site of a covalent bond of T to Z, Zs or Zo and Z, Zs and Zo are defined above for formulae I, II and III, respectively, and wherein (X) is the site of a covalent bond of T to X, X or X and X, X and X are defined above for formula I, II and III, respectively, L, is —CH2— or —O—: U is —CRoRo or —C(=O)—, Roo is lower alkyl; Ro and Ro are each independently hydrogen, lower alkyl or substituted lower alkyl; XX is 2 or 3; yy is 1 or 2: ZZ is 1 or 2; and aaa is 0 or 1. (0255 C. Solid Phase Techniques 0256 Specific solid phase techniques for the synthesis of the macrocyclic compounds of the invention have been described in WO 01/25257, WO 2004/111077, WO 2005/ 012331 and WO 2005/012332. Solution phase synthesis routes, including methods amenable to larger scale manufac ture, were described in U.S. Provisional Patent Application Ser. Nos. 60/622,055 and 60/642,271. 0257. In certain cases, however, the lability of protecting groups precluded the use of the standard basic medium for cyclization in the thioester Strategy discussed above. In these cases, either of two acidic methods was employed to provide macrocyclization under acid conditions. One method utilized HOAc, while the other method employed HOAt (Scheme 2). For example, the acetic acid cyclization was used for com pound 219. 0258. After executing the deprotection of the Ddz or Boc group on the tether, the resin was washed sequentially with DCM (2x), DCM-MeOH (1:1, 2x), DCM (2x), and DIPEA DCM (3:7, 1x). The resin was dried under vacuum for 10 min, then added immediately to a solution of HOAc in degassed DMF (5% v/v). The reaction mixture was agitated at 50-70° C., O/N. The resin was filtered, washed with THF, and the combined filtrate and washes evaporated under reduced pres Sure (water aspirator, then oil pump) to afford the macrocycle.
Scheme 2: Alternative Cyclization Methodologies 2% TFA, 3% TES Ddz-Tether-(Bts)-AA-AA-AA3 - - CFCOHN-Tether-(Bts)-AA-AA-AA DCM, rt, 1 h Resin wash 2xDCM, 2x(DCM-MeOH), 2x DCM (all for 5 min) and 3:7 DIPEA-DCM (for 3 min); dried quickly used immediately US 2011/02881 63 A1 Nov. 24, 2011 26
-continued HOAt (2 eq)/DMF (2.5 mL, degassed) 5% 0. AcOHDMF (2.5 mL, degassed) 50° C., 16 h 50-70° C., 16h
Macrocycle Macrocycle
0259 For a representative macrocycle with tether T, the method used for attachment of the tether, either a Mit AA-Leu, AA=Leu, AA=Phe, the application of the HOAt sunobu reaction (previously described in WO 01/25257) or method shown in Scheme 2 provided the cyclic peptidomi reductive amination (previously described in WO 2004/ 111077). The relevant deprotection and coupling protocols as metic in 10% yield, while the acetic acid method was more appropriate for the nature of the building block are performed effective, and gave 24% overall yield of the same macrocycle. utilizing standard procedures and those described in WO This latter methodology was particularly effective for com 2004/111077 for the assembly of the cyclization precursors. pounds containing His(Mts) residues. For example, with The building blocks are listed in the opposite order from tether T8. AA=Phe, AAAcp, AA=His(Mts), the macro which they are added in order to correlate the building block cycle was obtained in 20% overall yield, although the major number with standard peptide nomenclature. Hence BB is ity of the product no longer had the Mts group on histidine added first, followed by BB, then BB, finally the tether (T). (15:1 versus still protected). In the case of the RCM, the tether is not formed completely 0260 Synthesis of representative macrocyclic compounds until the cyclization step, but the portion of the tether attached of the present invention are shown in the Examples below. to BB is still added at this stage of the sequence, unless it is Table 1A below presents a summary of the synthesis of 224 already part of that building block. The final macrocycles are representative compounds of the present invention. The reac obtained after application of the appropriate deprotection tion methodology employed for the construction of the mac sequences. If any reaction was required to be carried out rocyclic molecule is indicated in Column 2 and relates to the post-cyclization, it is listed in Column 9. All of the macro particular scheme of the synthetic strategy, for example, use cycles presented in Table 1A were purified and met internal of the thioester strategy as shown in FIG. 2 or the RCM acceptance criteria. Yields (Column 10) are either isolated or approach as shown in FIG. 3. Column 3 indicates if any as calculated based upon CLND analysis. It should be noted Substituents are present on N. Columns 4-6 and 8 indicate that compounds 58 and 99 were not cyclized and represent the the individual building blocks employed for each compound, linear analogues of compounds 10 and 133, respectively. The amino acids, hydroxy acids or tether utilizing either standard lack of binding potency observed with these linear analogues nomenclature or referring to the building block designations illustrates the importance of the macrocyclic structural fea presented elsewhere in this application. Column 7 indicates ture for the desired activity.
TABLE 1A Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1 Attachment Additional Yield pound Method R BB BB, BBs Method Tether Reaction** (%)* 1 Thioester BtS-NIe Boc-Sar Boc-(D)Phe Mitsunobu Boc-T9 None 10.1 Strategy Reaction 2 Thioester BtS-Ile Boc-(D)Ala Boc-(D)Phe Mitsunobu Boc-T9 None 13.8 rategy Reaction 3 Thioester BtS-Val Boc-Sar Boc-(D)Phe Mitsunobu Boc-T9 None 10.3 rategy Reaction 4 Thioester BtS-Nva Boc Boc-(D)Phe Mitsunobu Boc-T9 None 4.6 rategy (D)NMeAla Reaction 5 Thioester BtS-Nva Boc-NEtGly Boc-(D)Phe Mitsunobu Boc-T9 None 8.6 rategy Reaction 6 Thioester BtS-Nva Ddz-Sar Ddz-(D)Trp(Boc) Mitsunobu DdZ-T9 None 8.1 rategy Reaction 7 Thioester BtS-Nva Ddz-Sar Ddz-(D)Tyr(But) Mitsunobu DdZ-T9 None 8.8 rategy Reaction 8 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 20.9 rategy Reaction 9 Thioester BtS-Val Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 9.7 rategy Reaction 10 Thioester BtS-Nva Boc-Sar Boc-(D)Phe Mitsunobu Boc-T9 None 9.9 rategy Reaction 11 Thioester BtS-Nva Boc-Sar Boc-(D)Phe Mitsunobu Boc-T8 None 9.9 rategy Reaction 12 Thioester Bts-(D)Val Boc-Nle Boc-Nle Mitsunobu Boc-T8 None 2.9 rategy Reaction 13 Thioester Bts-(D)Val Boc-Nva Boc-Phe Mitsunobu Boc-T8 None 5.8 Strategy Reaction US 2011/02881 63 A1 Nov. 24, 2011 27
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com Assembly NBB 1 Attachment Additional Yield pound Method BB BB BBs Method Reaction** (%)* 14 hioester BtS-Ile Boc-(D)Ala Boc-Phe Mitsunobu None 27.5 Strategy Reaction 15 hioester BtS-Ile Boc Boc-(D)Phe Mitsinobu None 19.5 rategy (D)NMeAla Reaction 16 hioester Bts-allo Boc Boc-(D)Phe Mitsunobu None 23.9 rategy le (D)NMeAla Reaction 17 hioester Bts-Cpg Boc Boc-(D)Phe Reductive None 24.8 S rategy (D)NMeAla Amination Reaction 18 hioester BtS-Acp Boc-Acp Boc-Phe Mitsunobu None 6.8 rategy Reaction 19 s hioester BtS-Val Boc Boc-Phe Mitsunobu None 12.7 rategy (D)NMeAla Reaction 2O hioester Bts-Leu Boc-Acp Boc-Phe(2-CI) Mitsunobu None 22.O rategy Reaction 21 hioester Bts-Leu Boc-Acp Boc-Phe(3-CI) Mitsunobu None 24.7 rategy Reaction 22 hioester Bts-Leu Boc-Acp Boc-1Nal Mitsunobu None 10.3 rategy Reaction 23 hioester Boc Boc-(D)Phe(2-CI) Mitsunobu None 32.6 rategy (D)NMeAla Reaction 24 hioester Boc Boc-(D)Phe(3-CI) Mitsunobu None 22.4 rategy (D)NMeAla Reaction 25 hioester Boc Boc-(D)Phe(4-CI) Mitsunobu None 21.0 rategy (D)NMeAla Reaction 26 hioester Boc Boc-(D)Phe(4-F) Mitsunobu None 15.5 rategy (D)NMeAla Reaction 27 hioester Boc Boc-(D)Tyr(OMe) Mitsunobu None 2O2 rategy (D)NMeAla Reaction 28 hioester Boc Boc-(D)Bip Mitsunobu None 31.6 rategy (D)NMeAla Reaction 29 hioester Boc Boc-(D)Dip Mitsunobu None 26.1 rategy (D)NMeAla Reaction 30 hioester Boc Boc-(D)1 Nal Mitsunobu None 31.9 rategy (D)NMeAla Reaction 31 hioester Boc Boc-(D)2Nal Mitsunobu None 21.9 rategy (D)NMeAla Reaction 32 hioester Boc Boc-(D)2Pal None 6.7 S rategy (D)NMeAla 33 Thioester Boc Boc-(D)4-ThzAla None 7.5 rategy (D)NMeAla 34 s hioester Boc Boc-(D)2-Thi None 14.2 rategy (D)NMeAla 35 hioester Boc Boc-(D)Phe None 9.4 rategy (D)NMeAla 36 hioester Boc Boc-(D)Phe None 13.0 Strategy (D)NMeAla 37 RCM Fmoc Fmoc-(D)Phe None 24.6 Strategy (D)NMeAla 38 RCM Fmoc Fmoc-(D)Phe Hydrogenation 44.2 Strategy (D)NMeAla 39 Thioester Boc Boc-(D)Phe None 21.4 Strategy (D)NMeAla 40 Thioester Boc Boc-(D)Phe None 18.6 rategy (D)NMeAla 41 hioester BtS-Ile Boc Boc-(D)Phe None 10.6 rategy (D)NMeAbu 42 hioester BtS-Tle Boc Boc-(D)Phe None 1.7 rategy (D)NMeAla 43 hioester BtS-Ile Boc Boc-(D)Phe None 0.4 rategy (D)NEtAla 44 hioester Bts-Leu Boc-Acp Boc-Phe None 7.8 rategy 45 hioester Bts-Leu Ddz-Glut OBut) None 11.6 rategy 46 hioester Bts-Leu Boc-Acp Boc-Val None 13.6 rategy 47 hioester Bts-Leu Boc-Acp Boc-Leu None 9.2 S rategy Reaction
US 2011/02881 63 A1 Nov. 24, 2011 29
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1- Attachment Additional Yield pound Method R BB BB- BBs Method Tether Reaction** (%)* 82 Thioester BtS-Nwa Boc-Sar Boc-(D)Phe Mitsunobu Boc-T3 None 3.9 Strategy Reaction 83 Thioester BtS-Nwa Boc-Sar Boc-(D)Phe Mitsunobu Boc-T16 None 1.8 rategy Reaction 84 Thioester BtS-Nwa Boc-Sar Boc-(D)Phe Mitsunobu Boc-T4 None 2.6 rategy Reaction 85 Thioester BtS-Nwa Boc-Sar Boc-(D)Phe Mitsunobu Boc-TS None 4.7 rategy Reaction 86 Thioester BtS-Nwa Boc-Sar Boc-(D)Phe Mitsunobu Boc-T14 None 0.4 rategy Reaction 87 Thioester Bts-Leu Boc-Acp Boc-Ala Mitsunobu Boc-T9 None 4.8 rategy Reaction 88 Thioester Bts-Leu Ddz-Acp Ddz-Tyr(But) Mitsunobu Ddz-T9 None 18.8 rategy Reaction 89 Thioester Bts-Leu Ddz-Acp Ddz-Trp(Boc) Mitsunobu Ddz-T9 None 16.5 rategy Reaction 90 Thioester Bts-Leu Boc-Acp Boc-Hfe Mitsunobu Boc-T9 None 8.5 rategy Reaction 91 Thioester Bts-Leu Ddz-Acp Ddz-Lys(Boc) Mitsunobu Ddz-T9 None 6.8 rategy Reaction 92 Thioester Bts-Leu Ddz-Acp Ddz-Glut OBut) Mitsunobu Ddz-T9 None 9.1 rategy Reaction 93 Thioester Bts-Leu Boc-Ala Boc-Phe Mitsunobu Boc-T9 None 9.2 rategy Reaction 94 Thioester Bts-Leu Boc-(D)Ala Boc-Phe Mitsunobu Boc-T9 None 21.8 rategy Reaction 95 Thioester Bts-Leu Boc-Aib Boc-Phe Mitsunobu Boc-T9 None 19.3 rategy Reaction 96 Thioester Bts-(D)Leu Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 7.0 rategy Reaction 97 Thioester Bts-Leu Boc-Acp Boc-(D)Phe Mitsunobu Boc-T9 None 9.2 rategy Reaction 98 Thioester Bts-(D)Leu Boc-Acp Boc-(D)Phe Mitsunobu Boc-T9 None 5.3 rategy Reaction 99 Thioester Ac Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T9 No cyclization 0.4 Strategy, Reaction linear OO Thioester Bts-Ala Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 0.4 rategy Reaction O1 Thioester BtS-Nle Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 9.0 rategy Reaction O2 Thioester BtS-Phe Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 5.8 rategy Reaction O3 Thioester Bts-Lys Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 2.9 rategy Reaction O4 Thioester BtS-Glu Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 9.3 rategy Reaction 05 Thioester BtS-Ser Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 1.9 rategy Reaction O6 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T3 None 6.3 rategy Reaction O7 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-TS None 4.2 rategy Reaction O8 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T12 None 18.3 rategy Reaction 09 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T11 None 10.1 rategy Reaction 10 Thioester Bts-Leu Boc-Acp Boc-Gly Mitsunobu Boc-T9 None 2.9 rategy Reaction 11 Thioester Bts-Leu Boc-Acc Boc-Phe Mitsunobu Boc-T9 None 3.0 rategy Reaction 12 Thioester Bts-Gly Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 3.2 rategy Reaction 13 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T9 None 16.9 rategy Reaction 14 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T16 None 2.9 rategy Reaction 15 Thioester Bts-Leu Boc-Acp Boc-Phe Mitsunobu Boc-T6 None O.S Strategy Reaction US 2011/02881 63 A1 Nov. 24, 2011 30
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1- Attachment Additional Yield pound Method R BB BB- BBs Method Tether Reaction** (%)* 16 Thioester Bts-Leu Ddz-Acp Ddz-GlucEt) Mitsunobu Ddz-T8 None 11.8 Strategy Reaction 17 Thioester BtS-Abu Boc- Boc-(D)Phe Mitsunobu Boc-T9 None 19.7 rategy (D)NMeAla Reaction 18 Thioester Bts-Leu Boc- Boc-(D)Phe Mitsunobu Boc-T9 None 21.0 rategy (D)NMeAla Reaction 19 Thioester BtS-Thr Boc- Boc-(D)Phe Mitsunobu Boc-T9 None 12.2 rategy (D)NMeAla Reaction 2O Thioester Bts- Boc- Boc-(D)Phe Reductive Boc-T9 None 17.5 Strategy Thr(OMe) (D)NMeAla Amination Reaction 21 Thioester BtS-Acc Boc- Boc-(D)Phe Mitsunobu Boc-T9 None 5.8 Strategy (D)NMeAla Reaction 22 Thioester Bts- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 22.1 rategy Phe(2-CI) Reaction 23 Thioester Bts- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 3.6 rategy Phe(3-CI) Reaction 24 Thioester Bts- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 9.8 rategy Phe(4-CI) Reaction 25 Thioester Bts- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 5.8 rategy Phe(4-F) Reaction 26 Thioester BtS-Hfe Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 9.8 rategy Reaction 27 Thioester Bts- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 4.5 rategy Tyr(OMe) Reaction 28 Thioester Bts-Bip Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 7.8 rategy Reaction 29 Thioester Bts-Dip Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 1.O rategy Reaction 30 Thioester BtS-1Nal Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 8.8 rategy Reaction 31 Thioester BtS-2Nal Boc-Acp Boc-Phe Mitsunobu Boc-T8 None S.O rategy Reaction 32 Thioester BtS-3Pal Boc-Acp Boc-Phe Reductive Boc-T8 None 7.0 Strategy Amination Reaction 33 Thioester BtS-4Pal Boc-Acp Boc-Phe Reductive Boc-T8 None 9.5 Strategy Amination Reaction 34 Thioester BtS-4- Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 2.0 Strategy ThzAla Reaction 35 Thioester BtS-2-Thi Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 4.0 rategy Reaction 36 Thioester BtS-Abu Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 3.3 rategy Reaction 37 Thioester BtS-Nwa Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 9.0 rategy Reaction 38 Thioester BtS-Ile Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 3.8 rategy Reaction 39 Thioester BtS-Val Boc-hcLeu Boc-Phe Reductive Boc-T8 None 8.4 Strategy Amination Reaction 40 Thioester BtS-Val Boc- Boc-Phe Reductive Boc-T8 None 6.7 Strategy hc(4O)Leu Amination Reaction 41 Thioester BtS-Val Boc- Boc-Phe Reductive Boc-T8 None 5.7 Strategy (4O)Acp Amination Reaction 42 Thioester BtS-Val Boc- Boc-Phe Reductive Boc-T8 None 7.0 Strategy (3-4)InAcp Amination Reaction 43 Thioester BtS-Val Boc- Boc-Phe Reductive Boc-T8 None 6.1 Strategy hc(4S) Leu Amination Reaction 44 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 5.7 Strategy (D)NMeVal Amination Reaction 45 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 4.9 Strategy NMeVal Amination Reaction US 2011/02881 63 A1 Nov. 24, 2011 31
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1- Attachment Additional Yield pound Method R BB BB- BBs Method Tether Reaction** (%)* 46 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 23.3 Strategy NMeNwa Amination Reaction 47 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 14.4 Strategy (D)NMeLeu Amination Reaction 48 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 25.4 Strategy NMeLeu Amination Reaction 49 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 11.4 Strategy (D)NMeIle Amination Reaction 50 Thioester BtS-Ile Boc- Boc-(D)Phe Reductive Boc-T9 None 7.0 Strategy NMeIle Amination Reaction 51 Thioester BtS-Ile Ddz- Boc-(D)Phe Mitsunobu Ddz-T9 None 8.2 Strategy (D)Ser(But) Reaction 52 Thioester BtS-Ile Ddz- Boc-(D)Phe Reductive Ddz-T9 None 22.1 Strategy NMeSer(But) Amination Reaction 53 Thioester Bts-Leu Boc-Acp Boc-Phe(4-CI) Mitsunobu Boc-T8 None 13.5 Strategy Reaction S4 Thioester Bts-Leu Boc-Acp Boc-Phe(4-F) Mitsunobu Boc-T8 None 14.4 rategy Reaction 55 Thioester Bts-Leu Boc-Acp Boc-Hfe Mitsunobu Boc-T8 None 13.5 rategy Reaction 56 Thioester Bts-Leu Boc-Acp Boc-Tyr(OMe) Mitsunobu Boc-T8 None 13.2 rategy Reaction 57 Thioester Bts-Leu Boc-Acp Boc-Bip Mitsunobu Boc-T8 None 2O2 rategy Reaction 58 Thioester Bts-Leu Boc-Acp Boc-Dip Mitsunobu Boc-T8 None 11.3 rategy Reaction 59 Thioester Bts-Leu Boc-Acp Boc-2Nal Mitsunobu Boc-T8 None 2O.S rategy Reaction 60 Thioester Bts-Leu Boc-Acp Boc-2Pal Reductive Boc-T8 None 2.8 Strategy Amination Reaction 61 Thioester Bts-Leu Boc-Acp Boc-3Pal Reductive Boc-T8 None 6.5 Strategy Amination Reaction 62 Thioester Bts-Leu Boc-Acp Boc-4Pal Reductive Boc-T8 None 6.7 Strategy Amination Reaction 63 Thioester Bts-Leu Boc-Acp Boc-4-ThzAla Mitsunobu Boc-T8 None O.O Strategy Reaction 64 Thioester Bts-Leu Boc-Acp Boc-2-Thi Mitsunobu Boc-T8 None 2.5 rategy Reaction 65 Thioester Bts-Leu Boc-Acp Boc-Abu Mitsunobu Boc-T8 None 3.0 rategy Reaction 66 Thioester Bts-Leu Boc-Acp Boc-Ile Mitsunobu Boc-T8 None 1.1 rategy Reaction 67 Thioester Bts-Leu Boc-Acp Boc-allo-Ile Mitsunobu Boc-T8 None 5.3 rategy Reaction 68 Thioester Bts-Leu Boc-Acp Boc-Acp Mitsunobu Boc-T8 None 4.2 rategy Reaction 69 Thioester BtS-Ile Boc- Boc-(D)Hfe Mitsunobu Boc-T9 None 7.0 rategy (D)NMeAla Reaction 70 Thioester BtS-Ile Boc- Boc-(D)3Pal Reductive Boc-T9 None 4.5 Strategy (D)NMeAla Amination Reaction 71 Thioester BtS-Ile Boc- Boc-(D)4Pal Reductive Boc-T9 None 6.4 Strategy (D)NMeAla Amination Reaction 72 Thioester BtS-Ile Boc- Boc-Abu Mitsunobu Boc-T9 None 2.0 Strategy (D)NMeAla Reaction 73 Thioester BtS-Ile Boc- Boc-(D)Nva Mitsunobu Boc-T9 None 6.8 Strategy (D)NMeAla Reaction US 2011/02881 63 A1 Nov. 24, 2011 32
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1- Attachment Additional Yield pound Method R BB BB- BBs Method Tether Reaction** (%)* 74 Thioester BtS-Ile Boc- Boc-(D)Val Mitsunobu Boc-T9 None 13.9 Strategy (D)NMeAla Reaction 75 Thioester BtS-Ile Boc- Boc-(D)Ile Mitsunobu Boc-T9 None 15.1 Strategy (D)NMeAla Reaction 76 Thioester BtS-Ile Boc- Boc-(D)Leu Mitsunobu Boc-T9 None 9.4 Strategy (D)NMeAla Reaction 77 Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Boc-T11 None 9.3 Strategy (D)NMeAla Reaction 78 Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Boc-T28 None 11.2 Strategy (D)NMeAla Reaction 79 Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Boc-T29 None 8.6 Strategy (D)NMeAla Reaction 8O Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Boc-T30 None 1O.O Strategy (D)NMeAla Reaction 81 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T-7 None 49.5 Strategy (D)NMeAla Reaction 82 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T-7 Hydrogenation 47.7 Strategy (D)NMeAla Reaction 83 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T-7 None 59.0 Strategy (D)NMeAla Reaction 84 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T-7 Hydrogenation 50.6 Strategy (D)NMeAla Reaction 85 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T None 12.4 Strategy (D)NMeAla Reaction 86 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T None 3.0 Strategy (D)NMeAla Reaction 87 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T None 30.9 Strategy (D)NMeAla Reaction 88 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2+ T None 34.9 Strategy (D)NMeAla Reaction 89 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T Hydrogenation 24.0 Strategy (D)NMeAla Reaction 90 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T Hydrogenation 32.5 Strategy (D)NMeAla Reaction 91 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2+ T None 32.2 Strategy (D)NMeAla Reaction 92 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T Hydrogenation 22.2 Strategy (D)NMeAla Reaction 93 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T+ T None 47.7 Strategy (D)NMeAla Reaction 94 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T + T Hydrogenation 23.7 Strategy (D)NMeAla Reaction 95 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe Mitsunobu T2 + T None 66.8 Strategy (D)NMeAla Reaction 96 Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Ddz-T32(Boc) None 13.0 Strategy (D)NMeAla Reaction 97 Thioester BtS-Ile Boc- Boc-(D)Phe Mitsunobu Ddz-TB1 (But) None 10.6 rategy (D)NMeAla Reaction 99 Thioester BtS-Val Boc-Acc Boc-Phe Reductive Boc-T8 None 16.O Strategy Amination Reaction 200 Thioester BtS-Val Boc-Acp Boc-Phe Mitsunobu Boc-T8 None 14.7 Strategy Reaction 2O1 Thioester Me BtS-Nwa Boc- Boc-(D)Phe Reductive Boc-T9 Reductive 32.4 Strategy (D)NMeAla Amination animation Reaction reaction with formaldehyde 2O2 Thioester Ac BtS-Nwa Boc- Boc-(D)Phe Reductive Boc-T9 Acetylation 14.2 Strategy (D)NMeAla Amination Reaction 2O3 Thioester Me Bts-Leu Boc-Acp Boc-Phe Reductive Boc-T8 Reductive 7.7 Strategy Amination animation Reaction reaction with formaldehyde 204 Thioester Ac Bts-Leu Boc-Acp Boc-Phe Reductive Boc-T8 Acetylation 11.5 Strategy Amination Reaction US 2011/02881 63 A1 Nov. 24, 2011 33
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Tether Com- Assembly NBB 1 Attachment Additional Yield pound Method R BB BB, BBs Method Tether Reaction** (%)* 205 Thioester BtS-Ile Boc Boc-(D)Abu Mitsunobu Boc-T None 19.9 Strategy (D)NMeAla Reaction 2O6 Thioester BtS-Ile Boc Boc-(D)Phe Mitsunobu Boc-T T34 None 26.2 rategy (D)NMeAla Reaction 2O7 Thioester BtS-Val Boc Boc-Phe Mitsunobu Boc-T None <1 rategy hc(4N)Leu Reaction 208 Thioester Bts-allo- Boc-Acp Boc-Phe Mitsunobu Boc-T None 16.7 rategy le Reaction 209 Thioester BtS-Ile Boc Boc-(D)allo-Ile Mitsunobu Boc-T None 8.6 rategy (D)NMeAla Reaction 210 Thioester BtS-2Pal Boc-Acp Boc-Phe Reductive Boc-T None 1.1 Strategy Amination Reaction 211 Thioester BtS-Val Boc Boc-Phe Reductive Boc-T None Strategy hc(4N)Leu Amination Reaction 212 Thioester BtS-Ile Boc-NMcAbu Boc-(D)Phe Mitsunobu Boc-T None 1.2 Strategy Reaction 213 Thioester BtS-Ile Boc-(D)4-Thz Boc-(D)Phe Reductive Boc-T None 1.O Strategy Amination Reaction 214 RCM Fmoc-Ile Fmoc Fmoc-(D)Phe Mitsunobu Hydrogenation 14.9 Strategy (D)NMeAla Reaction 215 isolated from synthesis of compound 151 216 Thioester BtS-Val Boc-Acc Boc-Phe Reductive Boc-T None 11.6 Strategy Amination Reaction 218 Thioester Bts-hcLeu Boc-Acp Boc-Phe Mitsunobu Boc-T None O.1 Strategy Reaction 219 Acetic Acid H Bts- Boc-Acp Boc-Phe Reductive Boc-T None 19.0 Cyclization His(Mts) Amination Reaction 220 Thioester BtS-Nwa Boc-Pro Boc-(D)Phe Mitsunobu Boc-T None 1S.O Strategy Reaction 221 Thioester BtS-Nwa Boc-(D)Pro Boc-(D)Phe Mitsunobu Boc-T None 14.9 rategy Reaction 222 Thioester Bts-Leu Boc-Pro Boc-Phe Mitsunobu Boc-T None 11.7 rategy Reaction 223 Thioester Bts-Leu Boc-(D)Pro Boc-Phe Mitsunobu Boc-T None 20.4 Strategy Reaction 224 RCM Fmoc-Ile Fmoc Fmoc-(D)Phe Mitsunobu Hydrogenation 8.2 Strategy (D)Hyp(But) Reaction 225 Thioester BtS-Pro Boc Boc-(D)Phe Reductive Boc-T None Strategy (D)NMeAla Amination Reaction 226 Thioester Bts-Pip Boc Boc-(D)Phe Reductive Boc-T None 13.5 Strategy (D)NMeAla Amination Reaction
*Overall Yield; based on theoretical resin loading, starting from ~500 mg resin **Additional reactions conducted post-cyclization, excpet where otherwise noted, to reach the desired product
0261 Table 1 B below presents a summary of the synthesis used for attachment of the tether. The building blocks are of 122 representative compounds of the present invention, listed in the opposite order from which they are added in order and Table 1C presents the synthesis of an additional 15 rep to correlate the building block number with standard peptide resentative compounds. For Table 1B, the reaction method ology employed for the construction of the macrocyclic mol nomenclature. Column 8 indicates if any additional reaction ecule is indicated in Column 2 and relates to the particular chemistry was applied, such as to remove auxiliary protection scheme of the synthetic strategy. Columns 3-6 indicate the or to reduce a double bond (as was performed with many individual building blocks employed for each compound, RCM intermediate products). All of the macrocycles in amino acids or tether utilizing either standard nomenclature Tables 1B and 1C were purified and met the acceptance or referring to the building block designations presented else criteria. Yields (Column 9-10) are either isolated or as calcu where in this application. Column 7 indicates the method lated based upon CLND analysis. US 2011/02881 63 A1 Nov. 24, 2011 34
TABLE 1B Synthesis of Representative Compounds of the Present Invention Macrocyclic Com Assembly Tether Additional Amount Yield pound Method BB, BB Attachment Reaction** (mg)* (%)* 298 hioester Bts-Cpg Boc Boc-(D)Phe(4-F) Mitsunobu None 29.7 12 Strategy (D)NMeAla Reaction 299 hioester Bts-Cpg Boc Boc-(D)Phe(4-CI) Mitsunobu None S4.1 17 rategy (D)NMeAla Reaction 301 hioester Boc-Acp Boc-Phe(3-CI) Mitsunobu None 36.S 10 rategy Tyr(But) Reaction 303 hioester Boc-Phe Mitsunobu None 60 16 rategy Reaction 305 hioester BtS-Ile Boc Boc-(D)His(Mts) Reductive None 110 31 S rategy (D)NMeAla Amination Reaction 306 hioester Boc Mitsunobu None 51 8 Strategy (D)NMeAla Reaction 307 RCM Fmoc Mitsunobu None 13.6 10 Strategy (D)NMeAla Reaction 3O8 hioester Boc Mitsunobu None 43.8 14 Strategy (D)NMeAla Reaction 309 hioester Boc Mitsunobu None 38.2 13 rategy (D)NMeAla Reaction hioester Boc Mitsunobu None 33.3 11 rategy (D)NMeAla Reaction hioester Boc Reductive None 18.6 S.1 S rategy (D)NMeAla Amination Reaction hioester Boc Boc-(D)Phe(2-F) Mitsunobu None 42.9 4 s rategy (D)NMeAla Reaction hioester Boc Boc-(D)Phe(3-F) Mitsunobu None 38.2 3 rategy (D)NMeAla Reaction hioester Bts-Cpg Boc Boc-(D)Phe(2,4-diCI) Mitsunobu None 39.7 2 rategy (D)NMeAla Reaction hioester Bts-Cpg Boc Boc-(D)Phe(3,4-diCI) Mitsunobu None 35.3 1 rategy (D)NMeAla Reaction hioester Bts-Cpg Boc Boc-(D)Phe(3,4-diF) Mitsunobu None 40.7 3 rategy (D)NMeAla Reaction hioester Bts-Cpg Boc Boc-(D)Phe(3,5-diF) Mitsunobu None 37.6 2 rategy (D)NMeAla Reaction hioester Boc Boc-(D)Phe(pentaF) Mitsunobu None 36.1 1 rategy (D)NMeAla Reaction hioester Bts-Cpg Boc Boc-(D)Phe(4-Br) Mitsunobu None 37.5 1 rategy (D)NMeAla Reaction 320 hioester Bts-Cpg Boc Boc-(D)Phe(4-I) Mitsunobu None 43.4 2 rategy (D)NMeAla Reaction 321 hioester Bts-Cpg Boc Boc-(D)Phe(4-CN) Mitsunobu None 34.5 1 rategy (D)NMeAla Reaction 322 hioester Boc Mitsunobu None 40.8 2 rategy (D)NMeAla Reaction 323 hioester Boc Boc-(D)Phe(3,4-diCMe) Mitsunobu None 27.3 8 rategy (D)NMeAla Reaction 324 hioester Boc Boc-(D)Trp Mitsunobu None 38.6 2 rategy (D)NMcAla Reaction 325 hioester Boc-Acp Boc-Phe(3-F) Mitsunobu None 33.7 O rategy Reaction 326 hioester Boc-Acp Boc-Phe(3-Br) Mitsunobu None 37.5 O rategy Reaction 327 hioester Boc-Acp Boc-Phe(3.S.-diF) Mitsunobu None 35.2 1 rategy Reaction 328 hioester Boc-Acp Boc-Phe(3-OMe) Mitsunobu None 31.5 O rategy Reaction 329 hioester Boc-Acp Boc-Phe(3-CN) Mitsunobu None 26.9 8 rategy Reaction 330 hioester Boc-Acp Boc-Phe(3,4-diCI) Mitsunobu None 38.4 1 rategy Reaction 331 hioester Boc-Acp Boc-Phe(3,4-diF) Mitsunobu None 37 1 rategy Reaction 332 hioester Boc-Acp Boc-Phe(3-CF) Mitsunobu None 30.6 9 rategy Reaction 333 hioester Boc-Acp Boc-3-Thi Mitsunobu None 49.6 8 rategy Reaction 334 hioester Boc-Aib Boc-Phe(3-CI) Mitsunobu None 32 1 S rategy Reaction US 2011/02881 63 A1 Nov. 24, 2011 35
TABLE 1 B-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Com- Assembly Tether Additional Amount Yield pound Method BB BB, BB Tether Attachment Reaction* (mg)* (%)* 335 Thioester Boc- Boc- Boc-(D)Phe(4-F) Boc-T9 Reductive None 62.2 18 Strategy Thr(OMe) (D)NMeAla Amination Reaction 336 Thioester Bts- Boc- Boc-(D)Phe(4-F) Boc-T9 Mitsunobu None 37.7 12 Strategy Ser(OMe) (D)NMeAla Reaction 337 Thioester Boc- Boc- Boc-(D)Phe(4-F) Boc-T9 Reductive Hydrogenolysis 67.5 7 Strategy Dap(Cbz) (D)NMeAla Amination Reaction 338 Thioester Bts- Boc- Boc-(D)Phe(4-F) Boc-T9 Mitsunobu None 60 2O Strategy Dab(Boc) (D)NMeAla Reaction 339 Thioester Bts- Boc- Boc-(D)Phe(4-F) Boc-T9 Mitsunobu None 63 2O rategy Orn(Boc) (D)NMeAla Reaction 340 Thioester Boc-Met Boc- Boc-(D)Phe(4-F) Boc-T9 Reductive None 14.4 4 Strategy (D)NMeAla Amination Reaction 341 Thioester BtS-3-Thi Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 48 14 Strategy Reaction 342 Thioester Bts- Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 37.7 10 rategy Phe(2-CN) Reaction 343 Thioester Bts- Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 91.3 25 rategy Phe(2-OMe) Reaction 344 Thioester Bts- Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 22.1 7 rategy Ser(OMe) Reaction 345 Thioester BtS-Ile Boc-(40)Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 48 13 rategy Reaction 346 Thioester Bts-Cpg Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 52.1 16 rategy Reaction 347 Thioester BtS-Ile Boc-Acp Boc-Ser(OBzl) Boc-T8 Mitsunobu None 17.1 6 rategy Reaction 348 Thioester BtS-Ile Boc-Acp Boc-Ser(OBzl) Boc-T8 Mitsunobu None 104.4 33 rategy Reaction 349 Thioester BtS-Aib Boc-Acp Boc-Phe(3-CI) Boc-T8 Mitsunobu None 23.6 7 rategy Reaction 350 Thioester BtS-Aib Boc-Aib Boc-Phe(3-CI) Boc-T8 Mitsunobu None 44 15 rategy Reaction 351 Thioester BtS-Acp Boc-(D)Ala Boc-Phe(3-CI) Boc-T8 Mitsunobu None 39.1 13 rategy Reaction 352 Thioester BtS-Acp Boc-Ala Boc-Phe(3-CI) Boc-T8 Mitsunobu None 15.7 5 Strategy Reaction 353 RCM Fmoc-Ile Fmoc- Fmoc-(D)Phe(4-F) T41 + TBA Mitsunobu None 47.8 25 Strategy (D)NMeAla Reaction 3S4 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T65 Mitsunobu None 26.8 9 Strategy (D)NMeAla Reaction 355 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T70 Mitsunobu None 36.8 12 rategy (D)NMeAla Reaction 356 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T72 Mitsunobu None 10 3 rategy (D)NMeAla Reaction 357 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Ddz-T74(Boc) Mitsunobu None 4.1.8 11 Strategy (D)NMeAla Reaction 358 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T41 + TBA Mitsunobu None 26.1 26 Strategy Reaction 359 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T58 Mitsunobu None 43.6 12 Strategy Reaction 360 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T.42 + TB6 Mitsunobu None 36.3 18 Strategy Reaction 361 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T.42 + TBA Mitsunobu None 36.3 32 Strategy Reaction 362 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T42 + Tel Mitsunobu Hydrogenation 59.4 57 Strategy Reaction 363 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T42 + TB7 Mitsunobu Hydrogenation 4.1.8 44 Strategy Reaction 364 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T42 + TB7 Mitsunobu Hydrogenation 49.1 51 Strategy Reaction 365 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) TA1+TB10 Mitsunobu Hydrogenation 31.2 35 Strategy Reaction 366 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T41 + TB7 Mitsunobu Hydrogenation 33.3 37 Strategy Reaction 367 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T33b Mitsunobu None 21.1 6 Strategy (D)NMeAla Reaction US 2011/02881 63 A1 Nov. 24, 2011 36
TABLE 1 B-continued Synthesis of Representative Compounds of the Present Invention Macrocyclic Com- Assembly Tether Additional Amount Yield pound Method BB BB, BBs Tether Attachment Reaction* (mg)* (%)* 368 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T33a Mitsunobu None 21.8 10 Strategy Reaction 369 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T9 Mitsunobu None 21.1 4 Strategy Reaction 370 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T.42 + TB6 Mitsunobu Hydrogenation 8.9 NA Strategy Reaction 371 RCM Fmoc-Ile Fmoc-Acp Fmoc-Phe(3-CI) T.42 + TBA Mitsunobu Hydrogenation 9.9 NA Strategy Reaction 372 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) BOC-T69 Mitsunobu None 30.9 10 Strategy (D)NMeAla Reaction 373 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T71 Mitsunobu None 34.9 11 rategy (D)NMeAla Reaction 374 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Ddz-T73 (Boc) Mitsunobu None 42.7 12 rategy (D)NMeAla Reaction 375 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T39 Mitsunobu None 22.3 7 rategy (D)NMeAla Reaction 376 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T40 Mitsunobu None 7.5 2 rategy (D)NMeAla Reaction 377 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T10 Mitsunobu None 14.6 5 rategy (D)NMeAla Reaction 378 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T58 Mitsunobu None 65.3 21 rategy (D)NMeAla Reaction 379 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T67 Mitsunobu None 36.3 12 rategy (D)NMeAla Reaction 380 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T66 Mitsunobu None 16.5 5 rategy Reaction 381 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T65 Mitsunobu None 22.5 7 rategy Reaction 382 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T70 Mitsunobu None 24.5 7 rategy Reaction 383 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T69 Mitsunobu None 25.2 7 rategy Reaction 384 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T71 Mitsunobu None 21.9 6 rategy Reaction 385 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T11 Mitsunobu None 23.3 7 rategy Reaction 386 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T39 Mitsunobu None 12 4 rategy Reaction 387 Thioester Bts-tle Boc-Acp Boc-Phe(3-CI) Boc-T68 Mitsunobu None 17.1 5 rategy Reaction 388 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T67 Mitsunobu None 30 9 rategy Reaction 389 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T68 Mitsunobu None 16.1 5 rategy (D)NMeAla Reaction 390 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T18 Mitsunobu None 28.7 10 rategy Reaction 391 Thioester Bts-Cpg Boc- Boc-(D)Phe(3,4,5-triF) Boc-T9 Mitsunobu None 45.4 14 rategy (D)NMeAla Reaction 392 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T40 Mitsunobu None 4.3 1 rategy Reaction 393 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T45 Mitsunobu None 2.1 1 rategy Reaction 394 Thioester Bts-Cpg Boc- Boc-(D)Phe(4-F) Boc-T38 Mitsunobu None 3.7 1 Strategy (D)NMeAla Reaction 395 RCM Fmoc-Ile Fmoc- Fmoc-Phe(3-CI) T41 + TB2 Mitsunobu Hydrogenation O.2 O.2 Strategy (4N)Acp Reaction 396 Thioester BtS-Acp Boc- Boc-Phe(3-CI) Boc-T8 Mitsunobu None 2.3 1 Strategy (D)NMeAla Reaction 397 Thioester BtS-Acp NMeAla Boc-Phe(3-CI) Boc-T8 Mitsunobu None 1.4 0.4 Strategy Reaction 398 RCM Bts-Cpg Boc- Boc-(D)Phe(4-F) T.42 + TB6 Mitsunobu Hydrogenation 3.8 1 Strategy (D)NMeAla Reaction 399 Thioester BtS-Ile Boc-Acp Boc-Phe(3-CI) Boc-T33b Mitsunobu None 5.7 4 Strategy Reaction
US 2011/02881 63 A1 Nov. 24, 2011
Method Chiral B: Grad40A-05 (Column: Chiralcel OD-RH, TABLE 2B-continued 0.46 cmx15 cm): Analytical Characterization for Representative (0273 1. Isocratic plateau of 40 min at 40% ACN, 60% of Compounds of the Present Invention a solution 50 mM of CHCOONH in H.O. (0274) 2.5 min gradient to 70% ACN, 30% of a solution 50 MW Calc MS (M + H)" mM of CHCOONH in H.O. Compound Molecular Formula (g/mol) Found (0275 3. Isocratic plateau of 10 min at 70% ACN, 30% of 415 C3OH39N4O4C 555.1 555 a solution 50 mM of CHCOONH in H.O. 417 C29H36N4O4FC 559.1 559 430 C3OH38N4O4FC 573.1 573 (0276 4.5 min gradient to 40% ACN, 60% of a solution 50 431 C31H44N4O4 536.7 537 mM of CHCOONH in H.O. 432 C31H43N4O4C 571.2 571 (0277 5. Isocratic plateau of 10 min at 40% ACN, 60% of Notes a solution 50 mM of CHCOONH in H.O. 1. Molecular formulas and molecular weights are calculated automatically from the structure 0278 6. Flow: 0.5 mL/min via ActivityBase software (DBS, Guildford, Surrey, UK). 2. M+H obtained from LC-MS analysis using standard methods. 0279 7. Column temperature: room temperature 3. Allanalyses conducted on material after preparative purification by the methods described above. 0280 8. Sample temperature: room temperature Method Chiral C: Grad 55A-05 (column: Chiralcel OD-RH, 0.46 cmx15 cm): TABLE C (0281 1. 40 min isocratic 55%/45% of ACN/50 mM Analytical Characterization for Representative CHCOONH in HO Compounds of the Present Invention (0282 2.5 min gradient to 70%/30% of ACN/50 mM MW Calc MS (M + H) CHCOONH in HO Compound Molecular Formula (g/mol) Found (0283 3. 10 min isocratic 70%/30% of ACN/50 mM CHCOONH in HO (0284. 4.5 min gradient to 55%/44% of ACN/50 mM CHCOONH in HO 439 C32B39N4OSC 595.1 595 0285 5. 10 min isocratic 55%/45% of ACN/50 mM 440 C37H47N4OSF 646.8 647 CHCOONH in HO 0286 6. Flow: 0.5 mL/min 442 C26H38N4OS 486.6 487 0287 7. Column temperature: room temperature 0288 8. Sample temperature: room temperature 446 C24H3SN4OSF 478.6 479 Method Chiral D: Grad Isol 00B 05 (Column: Chiralcel OD-RH, 0.46 cmx15 cm): 449 C31H42NSO4C 584.1 S84 0289 1. 40 min isocratic 27%/73% of ACN/50 mM CHCOONH in HO Notes 1. Molecular formulas and molecular weights are calculated automatically from the structure 0290 2.5 min gradient to 70%/30% of ACN/50 mM via ActivityBase software (DBS, Guildford, Surrey, UK). CHCOONH in HO 2. M+H obtained from LC-MS analysis using standard methods. 3. Allanalyses conducted on material after preparative purification by the methods described 0291 3. 10 min isocratic 70%/30% of ACN/50 mM above. CHCOONH in HO 0292 4.5 min gradient to 27%/73% of ACN/50 mM 0263. D. Chiral Purity Determination CHCOONH in HO 0264 General methods for the HPLC determination of Stereoisomeric purity were employed according to techniques 0293 5. 10 min isocratic 27%/73% of ACN/50 mM known to those skilled in the art and further optimized for the CHCOONH in HO compounds of the present invention. 0294 6. Flow: 0.5 mL/min Method Chiral A: Grad35A-05 (Column: Chiralcel AS-RH, 0295 7. Column temperature: room temperature 0.46 cmx15 cm): 0296 8. Sample temperature: room temperature 0265 1. Isocratic plateau of 40 min at 35% ACN, 65% of a 50 mM solution of CHCOONH in H.O. 3. Biological Methods 0266 2.5 min gradient to 70% ACN, 30% of a 50 mM 0297. The compounds of the present invention were evalu solution of CHCOONH in H.O. ated for their ability to interact at the human ghrelin receptor 0267 3. Isocratic plateau of 10 min at 70% ACN, 30% of utilizing a competitive radioligand binding assay, fluores a 50 mM solution of CH3COONH in H.O. cence assay or Aequorin functional assay as described below. 0268 4.5 min gradient to 35% ACN, 65% of a 50 mM Such methods can be conducted in a high throughput manner solution of CHCOONH in H.O. to permit the simultaneous evaluation of many compounds. 0298 Specific assay methods for the human (GHS-R1a), 0269 5. Isocratic plateau of 10 min at 35% ACN, 65% of swine and rat GHS-receptors (U.S. Pat. No. 6.242,199, Intl. a 50 mM solution of CHCOONH in H.O. Pat. Appl. Nos. WO97/21730 and 97/22004), as well as the 0270. 6. Flow: 0.5 mL/min canine GHS-receptor (U.S. Pat. No. 6,645,726), and their use 0271 7. Column temperature: room temperature in generally identifying agonists and antagonists thereof are 0272 8. Sample temperature: room temperature known. US 2011/02881 63 A1 Nov. 24, 2011 42
0299. Appropriate methods for determining the functional Compound Handling activity of compounds of the present invention that interact at 0314 Compounds were provided frozen on dry ice at a the human ghrelin receptor are also described below. Stock concentration of 10 mM diluted in 100% DMSO and stored at -80° C. until the day of testing. On the test day, A. Competitive Radioligand Binding Assay (Ghrelin Recep compounds were allowed to thaw at rt O/N and then diluted in tor) assay buffer according to the desired test concentrations. Under these conditions, the maximal final DMSO concentra 0300. The competitive binding assay at the human growth tion in the assay was 0.1%. hormone secretagogue receptor (hGHS-R1a) was carried out analogously to assays described in the literature. (Bednarek Assay Protocol M A et al. Structure-function studies on the new growth 0315. In deep-well plates, 220 uL of diluted cell mem hormone-releasing peptide ghrelin: minimal sequence of branes (final concentration: 0.71 ug/well) were combined ghrelin necessary for activation of growth hormone secreta with 40 uL of either binding buffer (total binding, N=5), 1 uM gogue receptor 1a; J. Med. Chem. 2000, 43, 4370-4376: ghrelin (non-specific binding, N=3) or the appropriate con Palucki, B. L. et al. Spiro (indoline-3,4'-piperidine) growth centration of test compound (N=2 for each test concentra tion). The reaction was initiated by addition of 40 uL of hormone secretagogues as ghrelin mimetics: Bioorg. Med. '*'Il-ghrelin (final conc. 0.0070-0.0085 nM) to each well. Chem. Lett. 2002, 11, 1955-1957.) Plates were sealed with TopSeal-A, Vortexed gently and incu bated at rt for 30 min. The reaction was arrested by filtering Materials samples through Multiscreen Harvest plates (pre-soaked in 0301 Membranes (GHS-R/HEK 293) were prepared 0.5% polyethyleneimine) using a Tomtec Harvester, washed from HEK-293 cells stably transfected with the human ghre 9 times with 500 uL of cold 50 mM Tris-HCl (pH 7.4, 4°C.), lin receptor (hGHS-R1a). These membranes were provided and then plates were air-dried in a fumehood for 30 min A by PerkinElmer BioSignal (HRBHGHSM, lotil 1887) and uti bottom seal was applied to the plates prior to the addition of lized at a quantity of 0.71 ug/assay point. 25uL of MicroScint-0 to each well. Plates were than sealed with TopSeal-A and counted for 30 sec per well on a Top 0302) 1. ''Il-Ghrelin (PerkinElmer, #NEX-388); final Count Microplate Scintillation and Luminescence Counter concentration: 0.0070-0.0085 nM (PerkinElmer) using a count delay of 60 sec. Results were 0303 2. Ghrelin (Bachem, HH-4864); final concentration: expressed as counts per minute (cpm). 1 uM 0316 Data were analyzed by GraphPad Prism (GraphPad 0304 3. Multiscreen Harvest plates-GF/C (Millipore, Software, San Diego, Calif.) using a variable slope non-linear regression analysis. K values were calculated using a K. 0305. 4. Deep-well polypropylene titer plate (Beckman value of 0.01 nM for ''Il-ghrelin (previously determined Coulter, #267006) during membrane characterization). (0306 5. TopSeal-A (PerkinElmer, #6005185) D, values were calculated using the following formula: 0307 6. Bottom seal (Millipore, iMATAHOP00) D=1-testex concentration with maximal displace 0308 7. MicroScint-0 (PerkinElmer, #6013.611) ment-non-specific bindingx100 total binding-non 0309 8. Binding Buffer: 25 mM Hepes (pH 7.4), 1 mM specific binding CaCl, 5 mM MgCl, 2.5 mM EDTA, 0.4% BSA where total and non-specific binding represent the cpm obtained in the absence or presence of 1 LM ghrelin, respec Assay Volumes tively. 0310 Competition experiments were performed in a 300 0317 Binding activity at the gherlin receptor for represen tative compounds of the present invention is shown below in ul filtration assay format. Table 3A through 3E. Compound structures for Tables 3A, 3B 0311 1. 220 uL of membranes diluted in binding buffer and 3D are presented with the various groups as defined for 0312 2.40 uL of compound diluted in binding buffer the general structure of formula I. For Tables 3B and 3D, in all 0313. 3. 40 uL of radioligand (III-Ghrelin) diluted in entries, m, n and pare 0; X, Z and Z are each NH. For Table binding buffer 3B. R is H for all entries. The tethers (T) are illustrated with Final test concentrations (N=1) for compounds of the present the bonding to X and Z as indicated. The compounds them invention: selves are shown for Tables 3C and 3E. Competitive binding curves for representative compounds 1, 2, 3, 4 and 25 are 10, 1, 0.5,0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.002, 0.001 uM. shown in FIG. 4.
Binding Activity at the Human Ghrelin Receptor for Compounds of the Invention
Compound X R R2 l R7
1 N H H O CH US 2011/02881 63 A1 Nov. 24, 2011 43
-continued
CH
CH
CHCH
10 CH
11 CH
12
13
14 US 2011/02881 63 A1 Nov. 24, 2011 44
-continued
15 CH
16 CH
17 CH
18
19 CH
19b diastereomer
H
21
22
23 CH
24 CH
25 CH
26 CH US 2011/02881 63 A1 Nov. 24, 2011 45
-continued 27 CH
28 CH
29 CH
30 CH
31 CH
32 CH
33 CH
34 CH
35 CH
36 CH
37a CH
diastereomer
38 H CH
39 CH US 2011/02881 63 A1 Nov. 24, 2011 46
-continued
40 CH
41 CH
42 CH
43 CHCH
44
45
46
47
48
49
50
51 US 2011/02881 63 A1 Nov. 24, 2011 47
-continued 52 N H H - / O Me
53 N H H - / O Me
S4 N H H - / O Me
55 N H H O Me
56 N H H O Me
57 N H H - / O Me
58 N Ae H - / O Me
59 N H H - / O Me
60 N H H - / O H
61 N H H - / O H
62 N H H - / O H
63 N H H - / O H
64 N H H - / O H
65 N H H - / O H US 2011/02881 63 A1 Nov. 24, 2011 48
-continued
66
67
68
69
70
71
72 C H 3 CH 73 CH
O 74 >n-n-x- NHH CH 75 CH
76 CH
77 s CH
78 CH
79 CH US 2011/02881 63 A1 Nov. 24, 2011 49
-continued CH 81 CH
82 CH
83 CH
84 CH
85 CH
86 CH
87
88
89
90
91
92 US 2011/02881 63 A1 Nov. 24, 2011 50
-continued
93
94
95
96
97
98
99
C H 3
101
102
103 NH
104
H US 2011/02881 63 A1 Nov. 24, 2011 51
-continued
105 N H H O H
OH
106 N H H O H
107 N H H O H
108 N H H O H
109 N H H O H
110 N H H O H
111 N H H O H
112 N H H H O H
113 N H H O H
114 N H H O H
115 N H H O H
116 N H H O H US 2011/02881 63 A1 Nov. 24, 2011 52
-continued 117 - / CH 118 --- CH 119 CH
--(O 120 CH
O --(/ 121 -wl. CH 122 -C 123 - C 124 (). 125 * () 126 / O 127 * () O US 2011/02881 63 A1 Nov. 24, 2011 53
-continued
128
129
130
131 ( . ) 132 133 AC 134
teN 135 t 136a N H - / 136b diastereomer 137 H - / US 2011/02881 63 A1 Nov. 24, 2011
-continued
138
139
140
141
142
143
144 CH
145a CH
145b diastereomer 146a H CH
146b diastereomer
147 H CH
148 CH
149 CH
15Oa. N H CH US 2011/02881 63 A1 Nov. 24, 2011 55
-continued 1SOb diastereomer
151 H
152a. CH
diastereomer
153 H
154
155
157
158
159
16Oa. US 2011/02881 63 A1 Nov. 24, 2011 56
-continued
16Ob diastereomer
161a. N H H O H
161b. diastereomer
162a N H H O H
162b diastereomer
163 N H H O H
164 N H H O H
16S N H H O H
166 N H H O H
167 N H H O H
168 N H H O H
169 N H H O CH
170 N H H O CH US 2011/02881 63 A1 Nov. 24, 2011
-continued
171 CH
172 CH
173 CH
174 CH
175 CH
176 CH
177 CH
178 CH
179 CH
18O CH
181 CH
182a N H CH
182b diastereomer
183 H CH US 2011/02881 63 A1 Nov. 24, 2011
-continued 184 CH
184 diastereomer 185 H CH
186 CH
187 CH
188 CH
189a CH
189b diastereomer 190 H CH
191 CH
192 CH
193 CH
194a CH
194b. diastereomer 195 H CH
196 CH US 2011/02881 63 A1 Nov. 24, 2011
-continued 197 CH
199
CH
CH
204
205 CH
CH
208b. diastereomer
209 H CH
210 / US 2011/02881 63 A1 Nov. 24, 2011 60
-continued
211
212 CH
213
214 CH
215
216
218
219
220 See Figure 7A 221 See Figure 7A 222 See Figure 7A 223 See Figure 7A 224 See Figure 7A 225 See Figure 7B 226 See Figure 7B 227 See Figure 7B 228 See Figure 7B 229 See Figure 7B 230a See Figure 7B 23Ob diastereomer
Compound R
H CH CH US 2011/02881 63 A1 Nov. 24, 2011 61
-continued
10 11 t N t 12
13
14 CH 15 CH 16 CH 17 CH 18 O
19a CH 19b
21
22
23 24 25 26 27 28 29 30 31 32 33 34 35 36 37a 37b 38 C 13 O US 2011/02881 63 A1 Nov. 24, 2011 62
-continued
39 H O 40 CH N 41 H O
42 CH O 43 CH N O N— H
45
46
47
48
49 50 51 52 53 S4 55 56 57 58 59 60 CH 61
62 O
63 H O N H
64 H O N H US 2011/02881 63 A1 Nov. 24, 2011 63
-continued
65
66 CH CH 67
68
72 73 74 75 76 77 78 79 =#H 8O 81 82 83 84 85 86
87
88
89
90 US 2011/02881 63 A1 Nov. 24, 2011 64
-continued
91
92
93 H CH O N H 94 CH H O N H 95 CH CH O N H
96 O N— H
97
98
99
100
101
102
103
104
105 US 2011/02881 63 A1 Nov. 24, 2011 65
-continued
106
107
108
109
110
111
112
113
114
115
116
117 CH 118 CH 119 CH 120 CH 121 CH
122 US 2011/02881 63 A1 Nov. 24, 2011 66
-continued
123
124
125
126
127
128
129
130
131
132
133
134
135 US 2011/02881 63 A1 Nov. 24, 2011 67
-continued
136a Q- O N H
136b
137 Q- O N H
138 Q- O N H 139 ( ) O N H
140 A O N H 141 (-O O N H 142 O N H
143 ( ) O N H
144 O N H
145a H O N H
146a H - / O N H
146b US 2011/02881 63 A1 Nov. 24, 2011 68
-continued
147 H O N H
148 H O N H
149 H O N H
150a H O N H
1SOb
151 H O N H
OH
152a. H O N H
OH
152b
153
154
155
156
157
158 US 2011/02881 63 A1 Nov. 24, 2011 69
-continued
159
160a
16Ob
161a.
161b
162a
162b
163
164
16S
166
167
168
169 CH 170 CH 171 CH 172 CH 173 CH 174 CH 175 CH 176 CH 177 CH 178 CH 179 CH 18O CH
US 2011/02881 63 A1 Nov. 24, 2011
-continued
212 H O N H
213 H O N H
N N1 S 214 CH H O N H
215 H O N H
O
216 sh. O N H
218 O N H
219 O N H
220 221 222 223 224 225 226 227 228 229 230a 23Ob Compound Rs R p Z2
1 H O N H US 2011/02881 63 A1 Nov. 24, 2011 72
-continued
OH
10
11
12
13
14 US 2011/02881 63 A1 Nov. 24, 2011 73
-continued
15
16
17 HHH
18 H O N— H
19 H O N H
19b
2O H C O N H
21 H C O N H
22 H 8 O N H
23 C H O N H US 2011/02881 63 A1 Nov. 24, 2011 74
-continued
24 C H O N H
25 H O N H C
26 H O N H F
27 H O N H V 28 ( ) C H O N H
29 . H O N H
30 8 H O N H
31 ( ) H O N H
32 4 O H O N H
33
N US 2011/02881 63 A1 Nov. 24, 2011 75
-continued
34 H O N H
2
35
36
37a
38
39
40
41
42
43
44
US 2011/02881 63 A1 Nov. 24, 2011 77
-continued
58
59
60
61
62
63
64
65
66
67
68 US 2011/02881 63 A1 Nov. 24, 2011 78
-continued
69
70
71
72
73
74
75
76
77
78
79