USOO749 1695 B2
(12) United States Patent (10) Patent No.: US 7.491,695 B2 Fraser et al. (45) Date of Patent: *Feb. 17, 2009
(54) METHODS OF USING MACROCYCLIC S. Rudikoff, et al. Proc. Natl. Acad. Sci. USA (1982) 79, pp. 1979 MODULATORS OF THE GHRELIN 1983.* RECEPTOR J.-M. Cao et al. Trends Endocrin. Metab. (2006) 17(1), pp. 14-18.* Ahnfelt-Ronne et al. “Do Growth Hormone-Releasing Peptides Act (75) Inventors: Graeme L. Fraser, Québec (CA); as Ghrelin Secretagogues?” Endocrine 14(1): 133-135 (2001). Hamid R. Hoveyda, Québec (CA); Arcadi et al. “Electrophilic Cyclization of o-Acetoxy- and Mark L. Peterson, Québec (CA) o-Benzyloxyalkynylpyridines: An Easy Entry into 2,3-Disubsituted (73) Assignee: Tranzyme Pharma Inc., Quebec (CA) Furopyridines' Organic Letters 4(14): 2409-2412 (2002). Ariyasu et al. “Stomach is a Major Source of Circulating Ghrelin, and (*) Notice: Subject to any disclaimer, the term of this Feeding State Determines Plasma Ghrelin-Like Immunoreactivity patent is extended or adjusted under 35 Levels in Humans' The Journal of Clinical Endocrinology & U.S.C. 154(b) by 389 days. Metabolism 86(10): 4756-4758 (2001). Arvatet al. “Growth Hormone-Releasing Hormone and Growth Hor This patent is Subject to a terminal dis mone Secretagogue-Receptor Ligands' Endocrine 14(1): 35-43 claimer. (2001). (21) Appl. No.: 11/149,512 Backes et al. “Solid Support Linker Strategies' Current Opinion in Chemical Biology 1: 86-93 (1997). (22) Filed: Jun. 10, 2005 Baig et al. “Postoperative Ileus: A Review” Diseases of the Colon & Rectum 47: 516-526 (2002). (65) Prior Publication Data Baldanzi et al. “Ghrelin and des-acyl Ghrelin Inhibit Cell Death in US 2007/OO21331 A1 Jan. 25, 2007 Cardiomyocytes and Endothelial Cells through ERK1/2 and Pl 3-kinase/AKT The Journal of Cell Biology 159(6): 1029-1037 Related U.S. Application Data (2002). Baldwin et al. “Symbiotic Approach to Drug Design: (63) Continuation-in-part of application No. 10/872,142, Antihypertensive f3-Adrenergic Blocking Agents' Journal of Medici filed on Jun. 18, 2004. nal Chemistry 22(11): 1284-1290 (1979). (60) Provisional application No. 60/642.271, filed on Jan. Banks et al. “Extent and Direction of Ghrelin Transport Across the 7, 2005, provisional application No. 60/622,055, filed Blood-Brain Barrier Is Determined by its Unique Primary Structure” on Oct. 27, 2004, provisional application No. 60/621, The Journal of Pharmacology and Experimental Therapeutics 302: 642, filed on Oct. 26, 2004, provisional application No. 822-827 (2002). 60/479,223, filed on Jun. 18, 2003. Barreiro et al. “Developmental, State-Specific, and Hormonally (51) Int. Cl. Regulated Expression of Growth Hormone Secretagogue Receptor A6 IK 38/12 (2006.01) Messenger RNA in Rat Testis' Biology of Reproduction 68: 1631 CO7K 5/12 (2006.01) 1640 (2002). (52) U.S. Cl...... 514/9; 530/317 (Continued) (58) Field of Classification Search ...... None See application file for complete search history. Primary Examiner Andrew DKosar (56) References Cited (74) Attorney, Agent, or Firm Myers Bigel Sibley & U.S. PATENT DOCUMENTS Sajovec, PA 6,548,501 B2 4/2003 Hakkinen 6,586,403 B1* 7/2003 Mathison et al...... 514/18 (57) ABSTRACT 6,852,722 B2 2/2005 Hakkinen FOREIGN PATENT DOCUMENTS The present invention provides novel conformationally-de EP 1 159964 12/2001 fined macrocyclic compounds that have been demonstrated to WO WOO1/25257 4/2001 be selective modulators of the ghrelin receptor (growth hor WO WO 04/111077 12, 2004 mone secretagogue receptor, GHS-R1a and Subtypes, iso WO WO 05/O12331 2, 2005 forms and variants thereof). Methods of synthesizing the WO WO 05/O12332 2, 2005 novel compounds are also described herein. These com OTHER PUBLICATIONS pounds are useful as agonists of the ghrelin receptor and as medicaments for treatment and prevention of a range of medi W.-C. Qiu et al. J. Gastroenterol. (2008) 14(9), pp. 1419-1424.* J. Rudinger. In: Peptide Hormones, JA Parsons, Ed. (1976) 1-7.* cal conditions including, but not limited to, metabolic and/or D. Voet and J.G. Voet. Biochemistry, 2nd Edition.(1995), pp. 235 endocrine disorders, gastrointestinal disorders, cardiovascu 241.* lar disorders, obesity and obesity-associated disorders, cen D.E. Smilek, et al. Proc. Natl. Acad. Sci. USA (1991) 88, pp. 9633 tral nervous system disorders, genetic disorders, hyperprolif 9637.* erative disorders and inflammatory disorders. W.S. Messer, "Vasopressin and Oxytocin', web document updated Apr. 3, 2000;
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. Patent Feb. . 17, 2009 Sheet 3 of 21 US 7.491.695 B2
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qe!y U.S. Patent Feb. 17, 2009 Sheet 4 of 21 US 7.491.695 B2
Radioligand:[125I]GhrelinTarget: hCHS-R
Compound 1
-9 -8 -7 -6 -5 FIGURE 4A LOg M of Compound 1
Target: hCHS-R Radioligand: 125Ghrelin
50
Compound 2
LogM of compound? p FIGURE 4B U.S. Patent Feb. 17, 2009 Sheet 5 of 21 US 7.491.695 B2
Target: hCHS-R
Radioligand: (125)Ghrelin
Compound 3
LOg M of compounds t FIGURE 4C
Target: hCHS-R Radioligand: (125)Ghrelin
Compound 4 i
7 -6 -5 LOg M of compound 4. FIGURE 4D U.S. Patent Feb. 17, 2009 Sheet 6 of 21 US 7.491.695 B2
Target: hCHS-R Radioligand: (125)Ghrelin 100 g 75 50 3. 25 Compound 25
O 9 -8 -7 -6 -5 LOg M of Compound 25 FIGURE 4E
Compound 1
100
75 is 50
25 EC50 = 104 nM Hill slope = 1.3
-10 -9 -8 -7 -6 -5 -4 Log Concentration (M) FI GU R E 5A U.S. Patent Feb. 17, 2009 Sheet 7 of 21 US 7.491.695 B2
Compound 2 125
100 5075 2 5
O
-10 FIG URE 5B LOg Concentration (M)
Compound 3
100
5075 2 5 EC50 = 134 nM Hill slope = 1.9
-10 -9 -8 -7 -6 -5 -4 FIGU RE 5C Log Concentration (M) U.S. Patent Feb. 17, 2009 Sheet 8 of 21 US 7.491.695 B2
Compound 4 100
7 5
50
25
O
-10 -9 -8 -7 -6 -5 -4 FIGU RE5D LOg Concentration (M)
Compound 5
100
75
50
2 5 EC50=298 nM Hill slope = 1.5
-9 -8 -7 -6 -5 -4 FIGURESE LOg Concentration (M) U.S. Patent Feb. 17, 2009 Sheet 9 of 21 US 7.491,695 B2
400
T 3 O O
2 O O
1. O O
OH-I-I-I-I-I-T- O 60 120 180 24 O 300 360 FIGURE 6A Time (min)
BOO 700 600 500
3OO
3200O U 1 OO O O 3D 6O 90 12O 150 1 BO 21D 24 O 27O 3OO 33O 36O 390 FIGURE 6B Time (min) U.S. Patent Feb. 17, 2009 Sheet 10 of 21 US 7.491.695 B2
16DO 14OO 12DO OOO BOO DO OO 2OO O O 15 3O 45 SO 75 90 105 12O 135 15 FIGURE 6C Time (min)
SOO 1AOO 12OO 1 OOO BOO 6OO AOO 2OO O O 15 3O 45 6.O 75 90 105 12O 135 150 FIGURE 6D Time (min) U.S. Patent Feb. 17, 2009 Sheet 11 of 21 US 7.491.695 B2
O. 8
O. 6
O. 4.
O 2
O O o P C. 30 to 300 g C c 25 a 030Gr U. FIGURE 7A 39
OB vehicle metoclopramide (10 mg/kg)
GHRP-6 (20 pg/kg) Compound 298 (100 g/kg)
O) ) E 9 FIGURE 7B U.S. Patent Feb. 17, 2009 Sheet 12 of 21 US 7.491,.695 B2
Onawe POl+ vehicle
4 O to POI + Compound 298
2O
:
: 3.
ST 1D 20 3D 4 O 5 O SO 70 BO Georetiric Ceter of Gras? Naive - 35 cm PO - 6 cm Compound 298 - 16 cm FIGURE 8 U.S. Patent Feb. 17, 2009 Sheet 13 of 21 US 7.491.695 B2
300 Wehicle - 0.3 ml 225
15D
1OOO 11OO 12O 1300 14OO 15OO 1SOO Time (hours) FIGURE 9 U.S. Patent Feb. 17, 2009 Sheet 14 of 21 US 7.491.695 B2 Compound 298 Binding to h(GHS-R1a 100
7 5
5 O
2 5 Ki = 15 nM i Dmax = 96% Hill slope = 0.87
FIGURE to -10 Log Concentration-8 (M) -6 -4
100
75 v Compound 298 50 EC50 = 25 nM
Oghrelin EC50 = 2nM
1. 2 -10 -8 -6 FIGURE 11 LOGg Compound (M)(M U.S. Patent Feb. 17, 2009 Sheet 15 of 21 US 7.491.695 B2
Ghedin (5ug)
O
Ghrain (5ug)+ Compound 298 (300ugkg) 25
340
25
10 Compound 298
1OOO 105D 1100 1150 200 1250 1300 Time (hours FIGURE 12 U.S. Patent Feb. 17, 2009 Sheet 16 of 21 US 7.491.695 B2
FIGURE 13A
150- Compound 298
2 100- -- BS, A --- i-S
g: 50 A - . 10M
--. 5. ... 105M S O- 2 -i- - FIGURE 13B 1 to 3 3 ; ; ;
15 DeSensitization O Concentration response Curve Ghrein A GHRP-6 1. O O v Capromorelin Compound 298 5 O o Compound 430
O
5 O -1 -10 -9 -8 -7 -6 FIGURE 13C Pretreatment Concentration (LogM) U.S. Patent Feb. 17, 2009 Sheet 17 of 21 US 7.491.695 B2
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N N e. C O O o C O O N O O g w N ve wate (OI =U:OOlueNIOS) g-v=Uueeu |OJUOO UeNOSO 96 Ulueu00 peuOS US 7,491.695 B2 1. 2 METHODS OF USING MACROCYCLC ligand and is distinct from other receptors involved in the MODULATORS OF THE GHRELIN regulation of growth hormone (GH) secretion, in particular, RECEPTOR the growth hormone-releasing hormone (GHRH) receptor. A unique characteristic of both the rat and human peptides RELATED APPLICATION INFORMATION 5 is the presence of the n-octanoyl (Oct) moiety on Ser. How ever, the des-acyl form predominates in circulation, with This application is a continuation-in-part under 35 U.S.C. approximately 90% of the hormone in this form. This group is S 120 of U.S. patent application Ser. No. 10/872,142, filed derived from a post-translational modification and appears Jun. 18, 2004, currently pending, which claims the benefit relevant for bioactivity and possibly also for transport into the under 35 U.S.C. S 119(e) of U.S. Provisional Patent Applica 10 CNS. (Banks, W. A.; Tschöp, M.; Robinson, S. M.; Heiman, tion Ser. No. 60/479,223, filed Jun. 18, 2003. This continua M. L. Extent and direction of ghrelin transport across the tion-in-part application also claims the benefit under 35 blood-brain barrier is determined by its unique primary struc U.S.C. S 119(e) of U.S. Provisional Patent Application Ser. ture. J. Pharmacol. Exp. Ther: 2002, 302, 822-827.) In a No. 60/621,642, filed Oct. 26, 2004, U.S. Provisional Patent GH-releasing assay, the des-octanoyl form of the hormone Application Ser. No. 60/622,055, filed Oct. 27, 2004, and 15 was at least 100-fold less potent than the parent peptide, U.S. Provisional Patent Application Ser. No. 60/642,271, although it has been Suggested that the des-acyl species may filed Jan. 7, 2005. The disclosures of the above-referenced be responsible for some of the other biological effects asso applications are incorporated herein by reference in their ciated with ghrelin. This des-acyl form has also been postu entireties. lated to be primarily responsible for the cardiovascular and cell proliferation effects attributed to ghrelin, while the acy FIELD OF THE INVENTION lated form participates in maintenance of energy balance and growth hormone release. (Baldanzi, G.; Filighenddu, N.; The present invention relates to novel conformationally Cutrupi, S.; et al. Ghrelin and des-acyl ghrelin inhibit cell defined macrocyclic compounds that bind to and/or are func death in cardiomyocytes and endothelial cells through tional modulators of the ghrelin (growth hormone secreta 25 ERK 1/2 and PI-3-kinase/AKT.J. Cell Biol. 2002, 159, 1029 gogue) receptor including GHS-R1a and Subtypes, isoforms 1037) Similarly, des-Gln-ghrelin and its octanoylated and/or variants thereof. The present invention also relates to derivative have been isolated as endogenous forms of the intermediates of these compounds, pharmaceutical composi hormone arising from alternative splicing of the ghrelin gene, tions containing these compounds and methods of using the but both are found to be inactive in stimulating GH release in compounds. These novel macrocyclic compounds are useful 30 vivo. (Hosoda, H.; Kojima, M.; Matsuo, H., Kangawa, K. as therapeutics for a range of disease indications. In particu Purification and characterization of rat des-Gln'-ghrelin, a lar, these compounds are useful for treatment and prevention second endogenous ligand for the growth hormone secreta of gastrointestinal disorders including, but not limited to, gogue receptor. J. Biol. Chem. 2000, 275, 21995-2120.). post-operative ileus, gastroparesis, including diabetic gastro Other minor forms of ghrelin produced by post-translational paresis, opioid bowel dysfunction, chronic intestinal pseudo 35 processing have been observed in plasma, although no spe obstruction, short bowel syndrome and functional gas cific activity has been attributed to them. (Hosoda, H.; trointestinal disorders. Kojima, M., et al. Structural divergence of human ghrelin. Identification of multiple ghrelin-derived molecules pro BACKGROUND OF THE INVENTION duced by post-translational processing.J. Biol. Chem. 2003, 40 278,64-70.) The improved understanding of various physiological Even prior to the isolation of this receptor and its endog regulatory pathways enabled through the research efforts in enous peptide ligand, a significant amount of research was genomics and proteomics has begun to impact the discovery devoted to finding agents that can stimulate GH secretion. of novel pharmaceutical agents. In particular, the identifica The proper regulation of human GH has significance not only tion of key receptors and their endogenous ligands has created 45 for proper body growth, but also a range of other critical new opportunities for exploitation of these receptor/ligand physiological effects. GH and other GH-stimulating peptides, pairs as therapeutic targets. For example, ghrelin is a recently such as GHRH and growth hormone releasing factor (GRF), characterized 28-amino acid peptide hormone isolated origi as well as their derivatives and analogues, are administered nally from the stomach of rats with the orthologue subse via injection. Therefore, to better take advantage of these quently identified in humans. (Kojima, M., Hosoda, H. et al. 50 positive effects, attention was focused on the development of Nature 1999, 402, 656-660.) The existence of this peptide in orally active therapeutic agents that would increase GH secre a range of other species suggests a conserved and important tion, termed GH secretagogues (GHS). Additionally, use of role in normal body function. This peptide has been demon these agents was expected to more closely mimic the pulsatile strated to be the endogenous ligand for a previously orphan G physiological release of GH. protein-coupled receptor (GPCR), type 1 growth hormone 55 Beginning with the identification of the growth hormone secretatogue receptor (hGHS-R1a)(Howard, A. D.; Feighner, releasing peptides (GHRP) in the late 1970's, (Bowers, C. Y. S. D.; et al. A receptor in pituitary and hypothalamus that Growth hormone-releasing peptides: physiology and clinical functions in growth hormone release. Science 1996, 273, applications. Curr. Opin. Endocrinol. Diabetes 2000, 7, 168 974-977.) found predominantly in the brain (arcuate nucleus 174: Camanni, F. Ghigo, E.; Arvat, E. Growth hormone and Ventromedial nucleus in the hypothalamus, hippocampus 60 releasing peptides and their analogs. Front. Neurosci. 1998, and substantia nigra) and pituitary. (U.S. Pat. No. 6.242, 199: 19, 47-72; Locatelli, V., Torsello, A. Growth hormone secre Intl. Pat. Appl. Nos. WO97/21730 and WO 97/22004) The tagogues: focus on the growth hormone-releasing peptides. receptor has also been detected in other areas of the central Pharmacol. Res. 1997, 36, 415-423.) a host of agents have nervous system (CNS) and in peripheral tissues, for instance been studied for their potential to act as GHS. In addition to adrenal and thyroid glands, heart, lung, kidney, and skeletal 65 their stimulation of GH release and concomitant positive muscles. This receptor was identified and cloned prior to the effects in that regard, GHS were projected to have utility in isolation and characterization of the endogenous peptide the treatment of a variety of other disorders, including wast US 7,491.695 B2 3 4 ing conditions (cachexia) as seen in HIV patients and cancer al. A role for ghrelin in the central regulation of feeding. induced anorexia, musculoskeletal frailty in the elderly, and Nature 2001, 409, 194-198) For example, subjects suffering growth hormone deficient diseases. Many efforts over the with anorexia or bulimia exhibit elevated ghrelin levels. Cir past 25 years have yielded a number of potent, orally available culating levels of the hormone have been found to rise before GHS. (Smith, R. G.; Sun, Y. X., Beatancourt, L.; Asnicar, M. 5 meals and fall after meals. In addition, diet-induced weight Growth hormone secretagogues: prospects and pitfalls. Best loss leads to increased ghrelin levels, although obese Subjects Pract. Res. Clin. Endocrinol. Metab. 2004, 18, 333-347: who have gastric bypass Surgery do not likewise experience Fehrentz, J.-A.; Martinez, J.; Boeglin, D.; Guerlavais, V.: Such an increase. (Cummings, D. E.; Weigle, D. S.; Frayo, R. Deghenghi, R. Growth hormone secretagogues: Past, present S.; et al. Plasmaghrelin levels after diet-induced weight loss and future. IDrugs 2002, 5,804-814; Svensson, J. Exp. Opin. 10 or gastric bypass surgery. N. Engl. J. Med. 2002, 346, 1623 Ther. Patents 2000, 10, 1071-1080; Nargund, R. P.; Patchett, 1630) A. A. et al. Peptidomimetic growth hormone secretagogues. This intimate involvement of ghrelin in control of food Design considerations and therapeutic potential. J. Med. intake and appetite has made it an attractive target for obesity Chem. 1998, 41,3103-3127: Ghigo, E: Arvat, E.: Camanni, F. research. Indeed, few other natural substances have been Orally active growth hormone secretagogues: state of the art 15 demonstrated to be involved in the modulation of both GH and clinical perspective. Ann. Med. 1998, 30, 159-168; Smith, secretion and food intake. R. G.; Vander Ploeg, L. H. T.; Howard, A. D.; Feighner, S.D.: An additional effect of ghrelin that has not to date been et al. Peptidomimetic regulation of growth hormone secre exploited for therapeutic purposes is in modulating gastric tion. Endocr: Rev. 1997, 18, 621-645.) These include small motility and gastric acid secretion. The prokinetic activity peptides, such as hexarelin (Zentaris) and ipamorelin (Novo appears to be independent of the GH-Secretory action and is Nordisk), and adenosine analogues, as well as Small mol likely mediated by the Vagal-cholinergic muscarinic pathway. ecules such as carpomorelin (Pfizer), L-252,564 (Merck), The dose levels required are equivalent to those necessary for MK-0677 (Merck), NN7203 (Novo Nordisk), G-7203 (Ge the hormone's GH and appetite stimulation actions. It is nentech), S-37435 (Kaken) and SM-130868 (Sumitomo), noteworthy that, in contrast to its inactivity for ghrelin's other designed to be orally active for the stimulation of growth 25 actions, the des-Gln'' peptide demonstrated promotion of hormone. However, clinical testing with Such agents have motility as well. (Trudel, L.; Bouin, M.; Tomasetto, C. Eber rendered disappointing results due to, among other things, ling, P.; St-Pierre, S.; Bannon, P.; L'Heureux, M. C.; Poitras, lack of efficacy over prolonged treatment or undesired side P. Two new peptides to improve post-operative gastric ileus in effects, including irreversible inhibition of cytochrome P450 dog. Peptides 2003, 24, 531-534; Trudel, L.; Tomasetto, C.; enzymes (Zdravkovic M. Olse, A. K. Christiansen, T.; et al. 30 Rio, M. C.; Bouin, M.; Plourde, V.: Eberling, P: Poitras, P. Eur: J. Clin. Pharmacol. 2003, 58,683-688.) Therefore, there Ghrelin/motilin-related peptide is a potent prokinetic to remains a need for pharmacological agents that could effec reverse gastric postoperative ileus in rats. Am. J. Physiol. tively target the GHS-R1a receptor for therapeutic action. 2002, 282, G948-G952; Peeters, T. L. Central and peripheral Despite its involvement in GH modulation, ghrelin is pri mechanisms by which ghrelin regulates gut motility. J. marily synthesized in the oxyntic gland of the stomach, 35 Physiol. Pharmacol. 2003, 54(Supp. 4), 95-103.) although it is also produced in lesser amounts in other organs, Ghrelin also has been implicated in various aspects of including the kidney, pancreas and hypothalamus. (Kojima, reproduction and neonatal development. (Arvat, E., Gianotti, M.; Hsoda, H. Kangawa, K. Purification and distribution of L., Giordano, R., et al. Growth hormone-releasing hormone ghrelin: the natural endogenous ligand for the growth hor and growth hormone secretagogue-receptor ligands. Focus mone secretagogue receptor. Horm. Res. 2001, 56 (Suppl. 1), 40 on reproductive system. Endocrine 2001, 14, 35-43) Also of 93-97: Ariyasu, H.; Takaya, K.; Tagami. T.; et al. Stomach is significance are the cardiovascular effects of ghrelin, since a major source of circulating ghrelin, and feeding State deter the peptide is a powerful vasodilator. As such, ghrelin ago mines plasma ghrelin-like immunoreactivity levels in nists have potential for the treatment of chronic heart failure humans. J. Clin. Endocrinol. Metab. 2001, 86, 4753-4758) In (Nagaya, N.; Kangawa, K. Ghrelin, a novel growth hormone addition to its role in stimulating GH release, the hormone has 45 relasing peptide, in the treatment of chronic heart failure. a variety of other endocrine and non-endocrine functions Regul. Pept. 2003, 114, 71-77; Nagaya, N.; Kangawa, K. (Broglio, F. Gottero, C.; Arvat, E.: Ghigo, E. Endocrine and Ghrelin improves left ventricular dysfunction and cardiac non-endocrine actions of ghrelin. Horm. Res. 2003, 59, 109 cachexia in heart failure. Curr: Opin. Pharmacol. 2003, 3, 117) and has been shown to interact with a number of other 146-151; Bedendi, I., Alloatti, G.; Marcantoni, A.; Malan, D.; systems in playing a role in maintaining proper energy bal 50 Catapano, F: Ghé, C.; et al. Cardiac effects of ghrelin and its ance. (Horvath, T. L.; Diano, S.; Sotonyi, P.; Heiman, M.: endogenous derivatives des-octanoyl ghrelin and des-Gln Tschöp, M. Ghrelin and the regulation of energy balance—a ghrelin. Eur: J. Pharmacol. 2003,476, 87-95) Intl. Pat. Appl. hypothalamic perspective. Endocrinology 2001, 142, 4163 Publ. WO 2004/014412 describes the use of ghrelin agonists 4169; Casanueva, F. F.; Dieguez, C. Ghrelin: the link con for the protection of cell death in myocardial cells and as a necting growth with metabolism and energy homeostasis. 55 cardioprotectant treatment for conditions leading to heart Rev. Endocrinol. Metab. Disord. 2002, 3, 325-338). In par failure. Lastly, evidence has been obtained that ghrelin may ticular, ghrelin plays a role as an orexigenic signal in the have implications in anxiety and other CNS disorders as well control offeeding, in which it acts to counteract the effects of as the improvement of memory. (Carlini, V. P. Monzon, M. leptin. Indeed, it was the first gut peptide proven to have Such E. Varas, M. M., Cragnolini, A. B., Schioth, H. B., Sci orexigenic properties. (Kojima, M., Kangawa, K. Ghrelin, an 60 monelli, T. N. de Barioglio, S. R. Ghrelin increases anxiety orexigenic signaling molecule from the gastrointestinal tract. like behavior and memory retention in rats. Biochem. Bio Curr. Opin. Pharmacology 2002, 2, 665-668.) The hormone phys. Res. Commun. 2002, 299,739-743) also is implicated in the hypothalamic regulation of the Syn The myriad effects of ghrelin in humans have suggested the thesis and secretion of a number of other neuropeptides existence of subtypes for its receptor, although none have as involved in appetite and feeding behavior. Levels of ghrelin 65 yet been identified. (Torsello, A.; Locatelli, Y.; Melis, M. R.: are elevated in response to fasting or extended food restric Succu, S.; Spano, M. S.; Deghenghi, R.; Muller, E. E.; Argio tion. (Nakazato, M.; Murakami, N.; Date, Y.: Kojima, M.; et las, A.; Torsello, A.; Locatelli, V.; etal. Differential orexigenic US 7,491.695 B2 5 6 effects of hexarelin and its analogs in the rat hypothalamus: mone-releasing peptides ghrelin and growth hormone-releas indication for multiple growth hormone secretagogue recep ing peptide-6 with the motilin receptor in the rabbit gastric tor subtypes. Neuroendocrinology 2000, 72,327-332.) How antrum.J. Pharmacol. Exp. Ther: 2003, 305, 660-667.) ever, a truncated, inactive form of GHS-R1a, termed GHS The variety of activities associated with the ghrelin recep R1b, was isolated and identified during the original tor could also be due to differentagonists activating different characterization studies. Evidence is mounting that addi signaling pathways as has been shown forghrelin and adenos tional receptor subtypes could be present in different tissues ine, both of which interact as agonists at GHS-R1a (Carreira, to explain the diverse effects displayed by the endogenous M. C.: Camina, J. P.; Smith, R. G.; Casanueva, F. F. Agonist peptides and synthetic GHS. For instance, high affinity bind specific coupling of growth hormone secretagogue receptor ing sites for ghrelin and des-acyl ghrelin have also been found 10 type 1a to different intracellular signaling systems. Role of in breast cancer cell lines, cardiomyocytes, and guinea pig adenosine. Neuroendocrinology 2004, 79, 13-25.) heart that are involved in mediating the antiproliferative, car The functional activity of a GPCR has been shown to often dioprotective and negative cardiac inotropic effects of these require the formation of dimers or other multimeric com peptides. Similarly, specific GHS binding sites besides GHS plexes with itself or other proteins. (Park, P. S.; Filipek, S.; R1a and GHS-R1b have been found in prostate cancer cells. 15 Wells, J. W.; Palczewski, K. Oligomerization of G protein Further, ghrelin and des-acyl ghrelin exert different effects on coupled receptors: past, present, and future. Biochemistry cell proliferation in prostate carcinoma cell lines. (Cassoni, 2004, 43, 15643-15656: Rios, C. D.; Jordan, B.A., Gomes, I.: P.: Ghé, C.; Marrocco, T.; et al. Expression of ghrelin and Devi, L. A. G-protein-coupled receptor dimerization: modu biological activity of specific receptors for ghrelin and des lation of receptor function. Pharmacol. Ther: 2001, 92.71-87; acyl ghrelin in human prostate neoplasms and related cell Devi, L. A. Heterodimerization of G-protein-coupled recep lines. Eur: J. Endocrinol. 2004, 150, 173-184) These various tors: pharmacology, signaling and trafficking. Trends Phar receptor Subtypes may then be implicated independently in macol. Sci. 2001, 22,532-537.) Likewise, the activity of the the wide array of biological activities displayed by the endog ghrelin receptor might also be at least partially governed by enous peptides and synthetic GHS. Indeed, recently, the exist Such complexes. For example, certain reports indicate that ence of receptor Subtypes was offered as an explanation for 25 interaction of GHS-R1a with GHRH(Cunha, S. R.; Mayo, K. the promotion of fat accumulation by ghrelin, despite its E. Ghrelin and growth hormone (GH) secreatagogues poten potent stimulation of the lipolytic hormone, growth hormone. tiate GH-releasing hormone (GHRH)-induced cyclic adenos (Thompson, N. M.; Gill, D. A. S.; Davies, R.; Loveridge, N.; ine 3',5'-monophosphate production in cells expressing trans Houston, P. A.; Robinson, I. C. A. F.; Wells, T. Ghrelin and fected GHRH and GH secretagogue receptors. des-octanoyl ghrelin promote adipogenesis directly in vivo 30 Endocrinology 2002, 143, 4570-4582: Malagón, M. M.: by a mechanism independent of the type 1a growth hormone Luque, R. M.; Ruiz-Guerrero, E.; Rodríguez-Pacheco, F.; secretagogue receptor. Endocrinology 2004, 145, 234-242.) García-Navarro, S.: Casanueva, F. F.; Gracia-Navarro, F: Further, this work Suggested that the ratio of ghrelin and Castafio, J. P. Intracellular signaling mechanisms mediating des-acyl ghrelin production could help regulate the balance ghrelin-stimulated growth hormone release in Somatotropes between adipogenesis and lipolysis in response to nutritional 35 Endocrinology 2003, 144, 5372-5380) or between receptor Status. subtypes (Chan, C. B. Cheng, C. H. K. Identification and The Successful creation of peptidic ghrelin analogues that functional characterization of two alternatively spliced separate the GH-modulating effects of ghrelin from the growth hormone secretagogue receptor transcripts from the effects on weight gain and appetite provides strong evidence pituitary of black Seabream Acanthopagrus Schlegeli. Mol. for the existence and physiological relevance of other recep 40 Cell. Endocrinol. 2004, 214, 81-95) may be involved in tor subtypes. (Halem, H. A.; Taylor, J. E.; Dong, J. Z.; Shen, modulating the function of the receptor. Y.; Datta, R.; Abizaid, A.; Diano, S.; Horvath, T., Zizzari, P.; The vast majority of reported approaches to exploiting the Bluet-Pajot, M.-T.: Epelbaum, J.: Culler, M. D. Novel ana ghrelin receptor for therapeutic purposes have focused on logs of ghrelin: physiological and clinical implications. Eur: modulating metabolic functions. Similarly, the vast majority J. Endocrinol. 2004, 151, S71-S75.) BIM-28163 functions as 45 of literature on GHS focuses on conditions that can be treated an antagonist at the GHS-R1a receptor and inhibits receptor via its GH promoting actions. Some embodiments of the activation by native ghrelin. However, this same molecule is invention described herein, in particular, take advantage of a full agonist with respect to stimulating weight gain and food selective activation of the ghrelin receptor to provide an intake. Additionally, the existence of a still uncharacterized avenue for the treatment of diseases characterized by GI receptor Subtype has been proposed based on binding studies 50 dysmotility. The improved GI motility observed with ghrelin in various tissues that showed differences between peptidic demonstrates that ghrelin agonists may be useful in correct and non-peptidic GHS. (Ong, H.; Menicoll, N.; Escher. F.; ing conditions associated with reduced or restricted motility Collu, R.; Deghenghi, R.: Locatelli, V. Ghigo, E.; Muccioli, (Murray, C. D. R.; Kamm, M. A.; Bloom, S. R.; Emmanuel, G.; Boghen, M.: Nilsson, M. Endocrinology 1998, 139, 432 A.V. Ghrelin for the gastroenterologist: history and potential. 435.) Differences between overall GHS-R expression and 55 Gastroenterology 2003, 125, 1492-1502; Fujino, K., Inui, A.; that of the GHS-R1a subtype in rat testis have been reported. Asakawa, A.; Kihara, N.; Fujimura, M.; Fujimiya, M. Ghrelin (Barreiro, M. L.; Suominen, J. S.; Gaytan, F.; Pinilla, L.; induces fasting motor activity of the gastrointestinal tract in Chopin, L. K. Casanueva, F. F.; Dieguez, C.; Aguilar, E.; conscious fed rats. J. Physiol. 2003, 550, 227-240: Edholm, Toppari, J., Tena-Sempere, M. Developmental, stage-spe T.: Levin, F.; Hellström, P. M.; Schmidt, P.T. Ghrelin stimu cific, and hormonally regulated expression of growth hor 60 lates motility in the Small intestine of rats through intrinsic mone secretagogue receptor messenger RNA in rat testis. cholinergic neurons. Regul. Pept. 2004, 121, 25-30.) Biol. Reproduction 2003, 68, 1631-1640) A GHS-R subtype Included among these conditions is post-operative ileus on cholinergic nerves is postulated as an explanation for the (POI, Luckey, A.; Livingston, E.; Taché, Y. Mechanisms and differential actions of ghrelin and a peptidic GHS on neural treatment of postoperative ileus. Arch. Surg. 2003, 138, 206 contractile response observed during binding studies at the 65 214; Baig, M. K. Wexner, S.D. Postoperative ileus: a review. motilin receptor. (Depoortere, I. Thijs, T.: Thielemans, L.; Dis. Colon Rectum 2004, 47,516-526). POI is defined as the Robberecht, P.; Peeters, T. L. Interaction of the growth hor impairment of GI motility that routinely occurs following US 7,491.695 B2 7 8 abdominal, intestinal, gynecological and pelvic Surgeries. In vagus nerve and via direct prokinetic action at the gastric the U.S. alone, 4.3 million surgeries annually induce POI, mucosa. Moreover, a recent clinical study indicates that intra accounting for an economic impact of over S1 billion. POI is venous administration of the natural ghrelin peptide is an considered a deleterious response to Surgical manipulation effective acute therapy in diabetic gastroparesis patients. A with a variable duration that generally persists for 72 hours. It 5 ghrelin agonist would therefore be highly effective in over is characterized by pain, abdominal distention or bloating, coming the fundamental motility barrierfaced by gastropare nausea and Vomiting, accumulation of gas and fluids in the sis patients and correcting this condition. As with POI, no bowel, and delayed passage of stool. Patients are neither able accepted or efficacious therapy for diabetic gastroparesis is to tolerate oral feeding nor to have bowel movements until gut available and most current therapies aim to provide only function returns. POI leads to numerous undesirable conse 10 symptomatic relief. Further, many of the therapeutics in quences, including increased patient morbidity, the costly development have a mechanism of action similar to earlier prolongation of hospital stays and, further, is a major cause of products that have failed in this indication. Surgical proce hospital readmission. In addition, opiate drugs given as anal dures may ameliorate the disease process, but offer no possi gesics after Surgery exacerbate this condition due to their bility of cure. well-recognized side effect of inhibiting bowel function. 15 Opioid-induced bowel dysfunction (OBD, Kurz, A.; Sessler, D. J. Opioid-Induced Bowel Dysfunction. Drugs Surgical manipulation of the stomach or intestine causes a 2003, 63, 649-671.) is the term applied to the confluence of disorganization of the gut-brain signaling pathways, impair symptoms involving the reduced GI motility that results from ing GI activity and triggering POI. Ghrelin acts locally in the treatment with opioid analgesics. Approximately 40-50% of stomach to stimulate and coordinate the firing of vagal affer patients taking opioids for pain control experience OBD. It is ent neurons and thereby modulate gut motility. Thus, ghrelin characterized by hard, dry stools, straining, incomplete accelerates gastric emptying in humans (Inui, A.; Asakawa, evacuation, bloating, abdominal distension and increased A.; Bowers, C.Y.; Mantovani, G.; Laviano, A.; Meguid, M. gastric reflux. In addition to the obvious short-term distress, M.; Fujimiya, M. Ghrelin, appetite, and gastric motility: the this condition leads to physical and psychological deteriora emerging role of the stomach as an endocrine organ. FASEB.J. 25 tion in patients undergoing long term opioid treatment. Fur 2004, 18, 439-456; Peeters, T. L. Central and peripheral ther, the dysfunction can be so severe as to become a dose mechanisms by which ghrelin regulates gut motility. J. limiting adverse effect that actually prevents adequate pain Physiol. Pharmacol. 2003, 54(Supp. 4), 95-103.) and is a control. As with POI, a ghrelin agonist can be expected to potent agent proven to treat POI in animal models (Trudel, L.; counteract the dysmotility resulting from opioid use. Tomasetto, C.; Rio, M. C.; Bouin, M.; Plourde, V.; Eberling, 30 Two less common conditions may also be helped through P: Poitras, P. Ghrelin/motilin-related peptide is a potent pro the GI motility stimulation effects of ghrelin and ghrelin kinetic to reverse gastric postoperative ileus in rats. Am. J. agonists. Short bowel syndrome is a condition that occurs Physiol. 2002, 282, G948-G952: Trudel, L.; Bouin, M.: after resection of a Substantial portion of small intestine and is Tomasetto, C.; Eberling, P.; St-Pierre, S.; Bannon, P.; characterized by malnutrition. Patients are observed to have L'Heureux, M. C.; Poitras, P. Two new peptides to improve 35 decreased ghrelin levels resulting from loss of the ghrelin post-operative gastric ileus in dog. Peptides 2003, 24, 531 producing neuroendocrine cells of the intestine. It is possible 534). Ghrelin agonists duplicate the effects of ghrelin, thus the short bowel feeds back on the release of the hormone. targeting directly the underlying cause of POI to accelerate (Krsek, M.; Rosicka, M.: Haluzik, M.: et al. Plasmaghrelin normalization of gut function and enable more rapid dis levels in patients with short bowel syndrome. Endocr: Res. charge from the hospital. Intravenous administration is often 40 2002, 28, 27-33.) Chronic intestinal pseudo-obstruction is a the preferred route of treatment for POI due to the impaired syndrome defined by the presence of chronic intestinal dila GI motility in these patients that impedes oral therapy. No tion and dysmotility in the absence of mechanical obstruction agent is currently approved by the U.S. FDA specifically for or inflammation. Both genetic and acquired causes are known the treatment of POI. to result in this disorder, which affects high numbers of indi Another major motility disorder is gastroparesis, a particu 45 viduals worldwide annually. (Hirano, I.; Pandolfino, J. lar problem for both type I and type II diabetics. (Camilleri, Chronic intestinal pseudo-obstruction. Dig. Dis. 2000, 18, M. Advances in diabetic gastroparesis. Rev. Gastroenterol. 83-92.) Disord. 2002, 2, 47-56; Tack et al. Gastroenterology 2004; Other conditions and disorders that could be addressed 126: A485; Moreaux, B.; VandenBerg, J.; Thielmans, L.; through stimulation of the ghrelin receptor are: emesis Such Meulemans, A.; Coulie, B. Activation of the GHS receptor 50 as caused by cancer chemotherapy, constipation Such as asso accelerates gastric emptying in the dog. Digestive Disease ciated with the hypomotility phase of irritable bowel syn Week, 15-20 May 2004, New Orleans, La., USA Abstract drome (IBS), delayed gastric emptying associated with wast M1009; Tack et al. Gastroenterology 2004, 126: A74) Gas ing conditions, gastroesophageal reflux disease (GERD), troparesis (“stomach paralysis) is a syndrome characterized gastric ulcers (Sibilia, V.; Rindi, G.; Pagani, F.; Rapetti, D.; by delayed gastric emptying in the absence of any mechanical 55 Locatelli, V., Torsello, A.; Campanini, N.; Degenghi, R.; obstruction. It is variably characterized by abdominal pain, Netti, C. Ghrelin protects against ethanol-induced gastric nausea, vomiting, weight loss, anorexia, early satiety, malnu ulcers in rats: studies on the mechanism of action. Endocri trition, dehydration, gastroesophageal reflux, cramping and nology 2003, 144, 353–359.) and Crohn's disease. bloating. This chronic condition can lead to frequent hospi Additionally, GI dysmotility is a significant problem in talization, increased disability and decreased quality of life. 60 other mammals as well. For example, the motility dysfunc Severe, symptomatic gastroparesis is common in individuals tion termed ileus or colic is the number one cause of mortality suffering from diabetes, affecting from 5-10% of diabetics for among horses. Further, ileus is one of the most common a total patient population of 1 million in the U.S. alone. complications of equine intestinal Surgery, in other words, Neuropathy is a frequent, debilitating complication of diabe post-operative ileus. This condition may also have a non tes. Visceral neuropathy results in GI dysfunction, especially 65 Surgical etiology. Some horses may be predisposed to ileus involving the stomach, leading to impaired gastric motility. based upon the anatomy and functioning of their digestive Ghrelin promotes gastric emptying both by stimulating the tract. Virtually any horse is susceptible to colic with only US 7,491.695 B2 9 10 minor differences based upon age, sex and breed. Addition Despite this immense body of work, cyclic compounds ally, ileus may affect other animals, for example canines. have rarely been found to act at the receptor. When they have, (Roussel, A.J., Jr.; Cohen, N. D.; Hooper, R. N.; Rakestraw, antagonist activity has been more prevalent. For example, the P. C. Risk factors associated with development of postopera 14-amino acid compound, vapreotide, an SRIH-14 agonist tive ileus in horses. J. Am Vet. Med Assoc. 2001, 219, 72-78; 5 Van Hoogmoed, L. M.: Nieto, J. E.; Snyder, J. R.: Harmon, F. and Somatostatin mimetic, was demonstrated to be a ghrelin A. Survey of prokinetic use in horses with gastrointestinal antagonist. (Deghenghi R, Papotti M. Ghigo E. et al. Soma injury. Vet. Surg. 2004, 33,279-285.) to statin octapeptides (lanreotide, octreotide, Vapreotide, and Importantly, for most of the above conditions, no specific, their analogs) share the growth hormone-releasing peptide approved therapeutics exist and most therapies simply 10 receptor in the human pituitary gland. Endocrine 2001, 14. address symptomatic relief. However, specific modulation of 29-33.) The binding and antagonist activities of analogues of the ghrelin receptor provides an opportunity to directly target the site of pathophysiological disturbance to better treat the cortistatin, a cyclic neuropeptide known to bind nonselec underlying condition and improve clinical outcome. Further, tively to Somatostatin receptors, to the growth hormone secre unlike other agents that interact at the GHS-R1a receptor, the 15 tagogue receptor have been reported (Intl. Pat. Appl. WO compounds of the invention are believed not to stimulate 03/004518). (Deghenghi R, Broglio F, Papotti M, et al. Tar concurrent GH secretion. This separation of the gastrointes geting the ghrelin receptor—Orally active GHS and cortista tinal and GH effects has not previously been reported for any modulators of this receptor. However, as already mentioned, tin analogs. Endocrine 2003, 22, 13-18) In particular, one of the existence of analogues that separate the appetite control these analogues, EP-01492 (cortistatin-8) has been advanced and GH modulatory effects associated with ghrelin has been into preclinical studies for the treatment of obesity as a ghre recently reported. lin antagonist. These compounds exhibit an ICso of 24-33 nM. WO 01/00830 reports on short gastrointestinal peptides In addition, these cyclic compounds and their derivatives, (SGIP) that secrete growth hormone and also promote GI 25 plus their use with metal binding agents have been described motility, but these were not shown to be due to action at the ghrelin receptor. U.S. Pat. No. 6,548,501 discloses specific for their ability to be useful for radiodiagnostic or radiothera compounds, but as GHS, useful for stimulation of GI motility. peutic use in the treatment of tumors and acromegaly. Moreover, other endogenous factors are known to stimulate Cyclic and linear analogues of growth hormone 177-191 secretion of GH, but do not promote GI motility. Indeed, 30 have been studied as treatments for obesity (WO 99/12969), many actually inhibit this physiological function. Specific receptoragonists such as the compounds of the present inven with one particular compound, AOD9604, having entered the tion have much better potential to be selective and effective clinic for this indication. A compound already studied that is therapeutic agents. most similar to the molecules of the present invention is the Work has continued at the development of potent and selec 35 GHS, G-7203 (ECso-0.43 nM), the cyclic peptide analogue tive GHS with a number of small molecule derivatives now of the growth hormone releasing peptide, GHRP-2 (Elias, K. being known as has been recently Summarized. (Carpino, P. A.: Ingle, G. S.; Burnier, J. P.; Hammonds, G.; McDowell, R. Exp. Opin. Ther. Patents 2002, 12, 1599-1618.) Specific GHS are described in the following U.S. Pat. Nos. and Intl. Pat. S.; Rawson, T. E.; Somers, T.C.; Stanley, M.S.: Cronin, M.J. Appl. Publs. WO 89/07110; WO 89/07111; WO92/07578; 40 In vitro characterization of four novel classes of growth hor WO 93/04081; WO 94/11012: WO94/13696; WO94/19367; mone-releasing peptide. Endocrinol. 1995, 136,5694-5699). WO95/11029; WO95/13069; WO95/14666; WO95/17422: However, simplification of this cyclic derivative led to still WO95/17423; WO95/34311; WO 96/02530; WO96/15148; potent, linear compounds, whereas, for compounds of the WO 96/22996; WO 96/22997; WO 96/24580; WO 96/24587; invention, linear analogues have been found to be devoid of WO 96/32943; WO 96/33189, WO 96/35713; WO 96/38471; 45 WO 97/00894; WO97/06803; WO 97/07117; WO97/09060; ghrelin receptor activity. WO 97/11697; WO97/15191, WO 97/15573; WO97/21730; The macrocyclic compounds of the invention possess ago WO 97/22004: WO97/22367; WO 97/22620; WO97/23508; nist activity. As previously mentioned, however, unlike other WO 97/24369; WO97/34604; WO 97/36873; WO97/38709; agonists of the hCHS-R1a receptor, the compounds of the WO 97/40023; WO 97/40071; WO 97/41878; WO 97/41879; 50 WO 97/43278; WO 97/44042: WO 97/46252: WO98/03473; invention unexpectedly have an insignificant stimulatory WO 98/10653; WO 98/18815; WO 98/22124; WO98/46569; effect on the release of growth hormone. Accordingly, the WO 98/51687; WO98/58947; WO 98/58948: WO98/58949; compounds of the present invention can exhibit selective WO 98/58950; WO99/08697; WO99/09991; WO99/36431; action in the GI tract or for metabolic disorders without side WO 99/39730; WO99/45029; WO99/58501; WO99/64456; 55 effects due to GH release. WO99/65486, WO 99/65488; WO 00/01726; WO00/10975; WO 01/47558; WO 01/92292; WO 01/96300; WO 01/97831; U.S. Pat. No. 3,239,345; U.S. Pat. No. 4,036,979; U.S. Pat. SUMMARY OF THE INVENTION No. 4411,890; U.S. Pat. No. 5,492,916; U.S. Pat. No. 5,494, 919; U.S. Pat. No. 5,559, 128: U.S. Pat. No. 5,663,171; U.S. 60 The present invention provides novel conformationally Pat. No. 5,721,250; U.S. Pat. No. 5,721,251; U.S. Pat. No. defined macrocyclic compounds. These compounds can 5,723,616; U.S. Pat. No. 5,726,319; U.S. Pat. No. 5,767,124; function as modulators, in particular agonists, of the ghrelin U.S. Pat. No. 5,798,337; U.S. Pat. No. 5,830,433: U.S. Pat. (growth hormone secretagogue) receptor (GHS-R1a). No. 5,919,777; U.S. Pat. No. 6,034,216; U.S. Pat. No. 6,548, 501; U.S. Pat. No. 6,559,150; U.S. Pat. No. 6,576,686; U.S. 65 According to aspects of the present invention, the present Pat. No. 6,686,359; and U.S. Pat. Appl. Nos. 2002/0168343; invention relates to compounds according to formula I, II 2003/100494; 2003/130284; 2003/186844. and/or III: US 7,491.695 B2 11 12 heteroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, oXo, amino, halogen, formyl, acyl, carboxy, carboxyalkyl, (I) R4 carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, (CH2) O mercapto, Sulfinyl, Sulfonyl and Sulfonamido, or, alterna O R III III Y tively, Rs is a fused cycloalkyl, a Substituted fused cycloalkyl, N Z1 R6 a fused heterocyclic, a substituted fused heterocyclic, a fused 7 aryl, a substituted fused aryl, a fused heteroaryl or a substi Y- NR '', Rs tuted fused heteroaryl ring when substituted for hydrogen (CH2) atoms on two adjacent atoms; Pn,X such) 10 X is O. NR or N(R): 1.xy wherein R is hydrogen, lower alkyl, substituted lower T-Z alkyl, Sulfonyl, Sulfonamido or amidino and Rio is hydrogen, lower alkyl, or substituted lower alkyl, or alternatively R and R together form a 3-, 4-, 5-, 6- or or an optical isomer, enantiomer, diastereomer, racemate or 15 7-membered ring, optionally comprising an O. S or stereochemical mixture thereof, additional Natom in the ring, wherein the ring is option wherein: ally substituted with Rs as defined above: R is hydrogen or the side chain of an amino acid, or Z is O or NR, alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem wherein R is hydrogen, lower alkyl, or substituted lower bered ring, optionally comprising an O. S or Natom in the alkyl, or alternatively R and R or R and R together ring, wherein the ring is optionally Substituted with Rs as form a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, defined below, or alternatively R and R together form a 3-, optionally comprising an O. S or additional Natom in 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S the ring, wherein the ring is optionally Substituted with or additional Natom in the ring, wherein the ring is optionally Rs as defined above; substituted with Rs as defined below: 25 Z2 is O or NR2, wherein R is hydrogen, lower alkyl, or R is hydrogen or the side chain of an amino acid, or substituted lower alkyl: alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem m, n and pare each independently 0, 1 or 2; bered ring, optionally comprising an O. S or Natom in the T is a bivalent radical of formula IV: ring, wherein the ring is optionally Substituted with Rs as defined below; or alternatively RandR together form a 3-, 30 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S wherein dande are each independently 0, 1, 2, 3, 4 or 5:Y or additional Natom in the ring, wherein the ring is optionally and Z are each optionally present: U is —CRR - or substituted with Rs as defined below: —C(=O)—and is bonded to X of formula I; W. Yand R is hydrogen or the side chain of an amino acid, or Zare each independently selected from the group con alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem 35 bered ring, optionally comprising an O or Satom in the ring, wherein the ring is optionally substituted with Rs as defined below, or alternatively, R and R7 or R and R togetherform SO. , —CRRs , —CH=CH- with the configu a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally ration Zor E. —C=C- and the ring structures below: comprising an O.S or additional Natom in the ring, wherein 40 the ring is optionally substituted with Rs as defined below: R is hydrogen or the side chain of an amino acid, or alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem bered ring, optionally comprising an O or Satom in the ring, 45 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: 50 Rs and R are each independently hydrogen or the side chain of an amino 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: 55 R is hydrogen, lower alkyl, substituted lower alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, or a Substituted heterocyclic group, or alternatively R and R, 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 60 atom in the ring, wherein the ring is optionally Substituted with Rs as described below: wherein G and G2 are each independently a covalent Rs is substituted for one or more hydrogenatoms on the 3-, bond or a bivalent radical selected from the group 4-, 5-, 6-, 7- or 8-membered ring structure and is indepen dently selected from the group consisting of alkyl, Substituted 65 alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl, Substituted aryl, US 7,491.695 B2 13 14 —CH=CH with the configuration Z or E, and tuted aryl, heteroaryl, substituted heteroaryl, —C=C-, with G being bonded closest to the group hydroxy, alkoxy, aryloxy, oxo, amino, formyl, acyl, U; wherein any carbonatom in the rings not otherwise carboxy, carboxyalkyl, carboxyaryl, amido, carbam defined, is optionally replaced by N, with the proviso oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, that the ring cannot contain more than four Natoms; sulfonyl, sulfonamido and, only if a double bond is K. K. K. K. and Ks are each independently O, present to the carbon atom to which it is bonded, NR or S, wherein R is as defined below: halogen; and R and R are each independently hydrogen, lower Rao and Ra are each independently hydrogen, lower alkyl, or substituted lower alkyl, or alternatively R. alkyl, substituted lower alkyl, R as defined above, and R together form a 3- to 12-membered cyclic 10 or alternatively Rao and R together form a 3- to ring optionally comprising one or more heteroatoms 12-membered cyclic ring optionally comprising one selected from the group consisting of O, S and N, or more heteroatoms selected from the group consist wherein the ring is optionally substituted with Rs as ing of O, S and N wherein the ring is optionally defined above; substituted with Rs as defined above, or alternatively R2s, Rao and R2 are each independently hydrogen, 15 one of Rao and R is hydroxy, alkoxy, aryloxy, amino, alkyl, substituted alkyl, cycloalkyl, substituted mercapto, carbamoyl, amidino, ureido or guanidino, cycloalkyl, heterocyclic, Substituted heterocyclic, while the other is hydrogen, lower alkyl or substituted aryl, substituted aryl, heteroaryl, substituted het lower alkyl, except when the carbon to which Rao and eroaryl, formyl, acyl, carboxyalkyl, carboxyaryl, Rare bonded is also bonded to another heteroatom; amido, amidino, Sulfonyl or Sulfonamido; with the proviso that T is not an amino acid residue, dipep R and Rs are each independently hydrogen, lower tide fragment, tripeptide fragment or higher order peptide alkyl, Substituted lower alkyl, R, wherein R is a fragment comprising standard amino acids; side chain of an amino acid Such as a standard or unusual amino acid, or alternatively Ra and Rs togetherform a 3- to 12-membered cyclic ring option 25 H O ally comprising one or more heteroatoms selected R51, from the group consisting of O, S and N, or alterna ( tively one of R or Rs is hydroxy, alkoxy, aryloxy, O N HN H amino, mercapto, carbamoyl, amidino, ureido or V 1. R50 R53 Rs guanidino while the other is hydrogen, lower alkyl or 30 T2 substituted lower alkyl, except when the carbon to ŠYx1'nz O which R- and Rs are bonded is also bonded to another heteroatom; R2, R1, Ras and Ras are each optionally present and, when present, are substituted for one or more hydro 35 or an optical isomer, enantiomer, diastereomer, racemate or gen atoms on the indicated ring and each is indepen stereochemical mixture thereof, dently selected from the group consisting of halogen, wherein: trifluoromethyl, alkyl, substituted alkyl, cycloalkyl, Rso is —(CH), CH, —CH(CH)(CH), CH, Substituted cycloalkyl, a heterocyclic group, a Substi —(CH2)CH(CH), —C(CH), —(CHRss), Rs, or tuted heterocyclic group, aryl, Substituted aryl, het 40 —CH(ORs)CH, whereinss is 1, 2 or 3; tt is 1 or 2: uu is 0. eroaryl, Substituted heteroaryl, hydroxy, alkoxy, ary 1 or 2; and VV is 0, 1, 2, 3 or 4: Rss is hydrogen or C-C alkyl; loxy, amino, formyl, acyl, carboxy, carboxyalkyl, Rs is amino, hydroxy, alkoxy, cycloalkyl or Substituted carboxyaryl, amido, carbamoyl, guanidino, ureido, cycloalkyl; and Rs, is hydrogen, alkyl, acyl, amino acyl, amidino, cyano, nitro, mercapto, Sulfinyl, Sulfonyl Sulfonyl, carboxyalkyl or carboxyaryl; and Sulfonamido; 45 Rs is hydrogen, C-C alkyl or C-C alkyl substituted R27 is optionally present and is substituted for one or with hydroxy or alkoxy; more hydrogen atoms on the indicated ring and each Rs is —(CHRss), Rs.9, wherein ww is 0, 1, 2 or 3: Rss is is independently selected from the group consisting of hydrogen, C-C alkyl, amino, hydroxy oralkoxy; Rso is aryl, alkyl, substituted alkyl, cycloalkyl, substituted substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl cycloalkyl, a heterocyclic group, a Substituted hetero 50 or substituted cycloalkyl: cyclic group, aryl, Substituted aryl, heteroaryl, Substi Rs is hydrogen or C-C alkyl; tuted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, X is O. NR or N(R)"; amino, formyl, acyl, carboxy, carboxyalkyl, car wherein R is hydrogen, lower alkyl, substituted lower boxyaryl, amido, carbamoyl, guanidino, ureido, ami alkyl, Sulfonyl, Sulfonamido or amidino and Rio is dino, mercapto, Sulfinyl, Sulfonyl and Sulfonamido; 55 hydrogen, lower alkyl, or substituted lower alkyl: R2s, R29. Rso Rs.2, Ras Rs. Rs6 and Rs 7 are each Zs is O or NR2, wherein R is hydrogen, lower alkyl, or optionally present and, when no double bond is substituted lower alkyl; and present to the carbonatom to which it is bonded in the T is a bivalent radical of formula V: ring, two groups are optionally present, and when present, is Substituted for one hydrogen present in the 60 ring, or when no double bond is present to the carbon wherein dande are independently 0, 1,2,3,4 or 5:Y, and atom to which it is bonded in the ring, is substituted Z, are each optionally present, U is —CRRe— or for one or both of the two hydrogen atoms present on —C(=O)—and is bonded to X of formula II, wherein the ring and each is independently selected from the Reo and R are each independently hydrogen, lower group consisting of alkyl, Substituted alkyl, 65 alkyl, or substituted lower alkyl, or alternatively R and cycloalkyl, Substituted cycloalkyl, a heterocyclic R together form a 3- to 12-membered cyclic ring group, a Substituted heterocyclic group, aryl, Substi optionally comprising one or more heteroatoms selected US 7,491.695 B2 15 16 from the group consisting of O, S and N, wherein the substituted heterocyclic, aryl, substituted aryl, het ring is optionally substituted with Rs as defined above: eroaryl, Substituted heteroaryl, hydroxy, alkoxy, WY, and Z are each independently selected from the aryloxy, oxo, amino, formyl, acyl, carboxy, car boxyalkyl, carboxyaryl, amido, carbamoyl, guani dino, ureido, amidino, mercapto, Sulfinyl, Sulfonyl, sulfonamido and, only ifa double bond is present to the carbon atom to which it is bonded, halogen; with the configuration Z or E. —C=C- and the ring Ro is optionally present, and when present is substi structures depicted below: tuted for one or more hydrogen atoms on the ring 10 and each is independently alkyl, Substituted alkyl,
cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl. Sub stituted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, oxo, amino, formyl, acyl, 15 carboxy, carboxyalkyl, carboxyaryl, amido, car bamoyl, guanidino, ureido, amidino, mercapto, sulfinyl, sulfonyl or sulfonamido: K is O or S; and ff is 1, 2, 3, 4 or 5: with the proviso that T is not an amino acid residue, dipeptide fragment, tripeptide fragment or higher order peptide fragment comprising standard amino wherein G and G are as defined above, and wherein acids; any carbon atom in the ring is optionally replaced by O N, with the proviso that the aromatic ring cannot 25 contain more than four Natoms and the cycloalkyl ring cannot contain more than two Natoms; R is hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Sub stituted heterocyclic group, aryl, Substituted aryl, heteroaryl, substituted heteroaryl, formyl, acyl, carboxyalkyl, carboxyaryl, amido, amidino, sulfo nyl or Sulfonamido; Re and Rea are each independently hydrogen, lower alkyl, Substituted lower alkyl or R, or alterna tively R and R together form a 3- to 12-mem bered cyclic ring optionally comprising one or or an optical isomer, enantiomer, diastereomer, racemate or more heteroatoms selected from the group consist stereochemical mixture thereof, wherein: ing of O, S and N: or alternatively one of R and Ro is hydrogen, C-C alkyl or alternatively Ro and R, R is hydroxy, alkoxy, aryloxy, amino, mercapto, 40 together form a 4-, 5-, 6-, 7- or 8-membered ring, optionally carbamoyl, amidino, ureido or guanidino, while the comprising an O, N or Satom in the ring, wherein the ring is other is hydrogen, lower alkyl or substituted lower optionally substituted with Rs as defined below: alkyl, except when the carbon to which Re and Rea R7 is hydrogen, —(CH2)CH, —CH(CH) are bonded is also bonded to another heteroatom; (CH2)CH, -(CH2)CH(CH), —(CH) R-7 or and R indicates the side chain of an amino acid 45 —CH(OR77)CH or, alternatively R7 and Rio together form Such as a standard or unusual amino acid; a 4-, 5-, 6-, 7- or 8-membered ring, optionally comprising an Res and Rs are each optionally present, and, when O, N or S atom in the ring, wherein the ring is optionally present are substituted for one or more hydrogen substituted with Rs, as defined below; whereinaa is 0,1,2,3, atoms on the ring and each is independently halo 4 or 5: bb is 1, 2 or 3; cc is 0, 1, 2 or 3; and dd is 0, 1, 2, 3 or gen, trifluoromethyl, alkyl, Substituted alkyl, 50 4. R-7 is aryl, Substituted aryl, heteroaryl, Substituted het cycloalkyl, Substituted cycloalkyl, a heterocyclic eroaryl, cycloalkyl or substituted cycloalkyl; R-7, is hydrogen, group, a Substituted heterocyclic group, aryl. Sub alkyl, acyl, amino acyl, Sulfonyl, carboxyalkyl or car stituted aryl, heteroaryl, substituted heteroaryl, boxyaryl; hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car R7 is C-C alkyl, or alternatively R7 and R together boxy, carboxyalkyl, carboxyaryl, amido, carbam 55 form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris oyl, guanidino, ureido, amidino, cyano, nitro, mer ing an O or Satom in the ring, wherein the ring is optionally capto, Sulfinyl, Sulfonyl or Sulfonamido; substituted with Rs, as defined below: Re and R, are each optionally present, and when no R is hydrogen, or alternatively R7 and R, togetherform double bond is present to the carbonatom to which a 3-, 4-, 5-, 6- or 7-membered ring, optionally comprising an it is bonded in the ring, two groups are optionally 60 O, S or N atom in the ring, wherein the ring is optionally present, and, when present, each is Substituted for substituted with Rs, as defined below: one hydrogen present in the ring, or when no Ra is hydrogen or C-C alkyl or alternatively Ra and R-7s double bond is present to the carbonatom to which together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally it is bonded in the ring, is substituted for one or both comprising an O, N or Satom in the ring, wherein the ring is of the two hydrogen atoms present on the ring and 65 optionally substituted with Rs, as defined below: each is independently alkyl, Substituted alkyl, R7s is —(CHR-7s)R 79 or alternatively R-7s and R together cycloalkyl, Substituted cycloalkyl, heterocyclic, form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris US 7,491.695 B2 17 18 ing an O, N or Satom in the ring, wherein the ring is option cal dosage units comprising an effective amount of one or ally substituted with Rs as defined below; wherein Rs is more compounds of the present invention packaged with hydrogen, C-C alkyl, amino, hydroxy or alkoxy, and Rio is optional instructions for the use thereof. selected from the group consisting of the following struc Aspects of the present invention further provide methods tures: of Stimulating gastrointestinal motility, modulating GHS R1a receptor activity in a mammal and/or treating a gas trointestinal disorder comprising administering to a subject in need thereof an effective amount of a modulator that modu lates a mammalian GHS-R1a receptor. In particular embodi 10 ments, interaction of the modulator and the GHS-R1a recep tor does not result in a significant amount of growth hormone release. In still other embodiments, the modulator is a com pound of formula I, II and/or III. El Additional aspects of the present invention provide meth 15 ods of diagnosing tumors and/or acromegaly, comprising administering compounds of the present invention and a .." and radiolabeled metal binding agent and detecting the binding of 4 | / 5 | / the composition to a biological target, and treating tumors and/or acromegaly comprising administering a therapeuti cally effective amount of a composition comprising a com pound of the present invention. Further aspects of the present invention relate to methods wherein E. E. E. E. and Es are each optionally present of making the compounds of formula I, II and/or III. and when present are each independently selected from Aspects of the present invention further relate to methods the group consisting of halogen, trifluoromethyl, alkyl, 25 of preventing and/or treating disorders described herein, in Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a particular, gastrointestinal disorders, including post-opera heterocyclic group, a Substituted heterocyclic group, tive ileus, gastroparesis, Such as diabetic and post-Surgical aryl, substituted aryl, heteroaryl, substituted heteroaryl, gastroparesis, opioid-induced bowel dysfunction, chronic hydroxy, alkoxy, aryloxy, cyano, Sulfinyl, Sulfonyl and intestinal pseudo-obstruction, short bowel syndrome, emesis Sulfonamido, and represent Substitution at one or more 30 Such as caused by cancer chemotherapy, constipation Such as available positions on the monocyclic or bicyclic aro associated with the hypomotility phase of irritable bowel matic ring, wherein said substitution is made with the syndrome (IBS), delayed gastric emptying associated with same or different selected group member, and J and J wasting conditions, gastroesophageal reflux disease are each independently O or S; (GERD), gastric ulcers, Crohn's disease, gastrointestinal dis Rs. Rs, and Rs are eachindependently substituted for one 35 orders characterized by dysmotility and other diseases and or more hydrogen atoms on the 3-, 4-, 5-, 6-, 7- or 8-mem disorders of the gastrointestinal tract. bered ring structure and are independently selected from the The present invention also relates to compounds of formula group consisting of alkyl, Substituted alkyl, cycloalkyl, Sub I, II and/or III used for the preparation of a medicament for stituted cycloalkyl, a heterocyclic group, a Substituted hetero prevention and/or treatment of the disorders described herein. cyclic group, aryl, Substituted aryl, heteroaryl, Substituted 40 The foregoing and other aspects of the present invention heteroaryl, hydroxy, alkoxy, aryloxy, Oxo, amino, halogen, are explained in greater detail in the specification set forth formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, car below. bamoyl, guanidino, ureido, amidino, mercapto, Sulfinyl, Sul fonyl and Sulfonamido, or, alternatively, Rs. Rs, and Rs are BRIEF DESCRIPTION OF THE DRAWINGS each independently a fused cycloalkyl, a Substituted fused 45 cycloalkyl, a fused heterocyclic, a substituted fused hetero FIG. 1 shows a scheme presenting a general synthetic cyclic, a fused aryl, a Substituted fused aryl, a fused heteroaryl strategy to provide conformationally-defined macrocycles of or a substituted fused heteroaryl ring when substituted for the present invention. hydrogen atoms on two adjacent atoms; FIG. 2 shows a general thioester strategy for making mac X is O. NR or N(R): 50 rocyclic compounds of the present invention. wherein R is hydrogen, lower alkyl, substituted lower FIG. 3 shows a general ring-closing metathesis (RCM) alkyl, Sulfonyl, Sulfonamido or amidino and Rio is strategy for macrocyclic compounds of the present invention. hydrogen, lower alkyl, or substituted lower alkyl: FIG. 4 (panels A through E) shows competitive binding Zo is O or NR, wherein R is hydrogen, lower alkyl, or 55 curves for binding of exemplary compounds of the present substituted lower alkyl; and invention to the hCHS-R1a receptor. T is the same as defined for T with the exception that U. FIG.5 (panels A through E) shows concentration-response is bonded to X of formula III. curves for activation of the hCGHS-R1a receptor by exemplary According to further aspects of the present invention, the compounds of the present invention. compound is a ghrelin receptor agonist or a GHS-R1a recep 60 FIG. 6 shows graphs depicting pharmacokinetic param toragonist. eters for exemplary compounds of the present invention, spe Further aspects of the present invention provide pharma cifically after oral administration of 8 mg/kg compound 298 ceutical compositions comprising: (a) a compound of the (panel A), after Subcutaneous injection of 2 mg/kg compound present invention; and (b) a pharmaceutically acceptable car 298 with cyclodextrin (panel B), after intravenous adminis rier, excipient or diluent. 65 tration of 2 mg/kg compound 25 with cyclodextrin (panel C) Additional aspects of the present invention provide kits and after intravenous administration of 2 mg/kg compound comprising one or more containers containing pharmaceuti 298 with cyclodextrin (panel D). US 7,491.695 B2 19 20 FIG. 7 (panels A and B) shows graphs presenting effects on of 1, 2 or 3 double or triple bonds, or a combination of the two. gastric emptying for exemplary compounds of the present Such alkyl groups may also be optionally Substituted as invention. described below. FIG. 8 shows a graph presenting effects on postoperative When a subscript is used with reference to an alkyl or other illeus for an exemplary compound of the present invention. 5 hydrocarbon group defined herein, the subscript refers to the FIG. 9 (panels A through D) shows graphs depicting the number of carbon atoms that the group may contain. For effect on pulsatile growth hormone release for an exemplary example, C-C alkyl indicates an alkyl group with 2, 3 or 4 compound of the present invention. carbon atoms. FIG. 10 shows a competitive binding curve for binding of The term “cycloalkyl refers to saturated or partially unsat an exemplary compound of the present invention to the 10 urated cyclic hydrocarbon groups having from 3 to 15 carbon atoms in the ring, in some instances 3 to 7, and to alkyl groups hCHS-R1a receptor. containing said cyclic hydrocarbon groups. Examples of FIG. 11 shows an activation curve demonstrating the ago cycloalkyl groups include, but are not limited to, cyclopropyl. nism of an exemplary compound of the present invention. cyclopropylmethyl, cyclopentyl, 2-(cyclohexyl)ethyl, cyclo FIG. 12 shows a graph depicting the lack of effect on 15 heptyl, and cyclohexenyl. Cycloalkyl as defined herein also ghrelin-induced growth hormone release for an exemplary includes groups with multiple carbon rings, each of which compound of the present invention. may be saturated or partially unsaturated, for example deca FIG. 13 shows graphs depicting receptor desentization linyl, 2.2.1-bicycloheptanyl or adamantanyl. All Such associated with binding of exemplary compounds of the cycloalkyl groups may also be optionally substituted as present invention to the hGHS-R1a receptor. described below. FIG. 14 (panels A and B) shows graphs presenting effects The term “aromatic” refers to an unsaturated cyclic hydro on gastric emptying for an exemplary compound of the carbon group having a conjugated pi electron system that present invention. contains 4n+2 electrons where n is an integer greater than or FIG. 15 shows a graph presenting effects on postoperative equal to 1. Aromatic molecules are typically stable and are illeus for an exemplary compound of the present invention. 25 depicted as a planar ring of atoms with resonance structures FIG. 16 shows graphs depicting reversal of morphine-de that consist of alternating double and single bonds, for layed gastric emptying (panel A) and morphine-delayed gas example benzene or naphthalene. trointestinal transit (panel B) for an exemplary compound of The term “aryl” refers to an aromatic group in a single or the present invention. fused carbocyclic ring system having from 6 to 15 ring atoms, FIG.17 (panels A and B) shows graphs depicting effects on 30 in some instances 6 to 10, and to alkyl groups containing said gastroparesis for exemplary compounds of the present inven aromatic groups. Examples of aryl groups include, but are not tion. limited to, phenyl, 1-naphthyl 2-naphthyl and benzyl. Aryl as defined herein also includes groups with multiple aryl rings DETAILED DESCRIPTION which may be fused, as in naphthyl and anthracenyl, or 35 unfused, as in biphenyl and terphenyl. Aryl also refers to The foregoing and other aspects of the present invention bicyclic or tricyclic carbon rings, where one of the rings is will now be described in more detail with respect to other aromatic and the others of which may be saturated, partially embodiments described herein. It should be appreciated that unsaturated or aromatic, for example, indanyl or tetrahy the invention can be embodied in different forms and should dronaphthyl (tetralinyl). All Such aryl groups may also be not be construed as limited to the embodiments set forth 40 optionally substituted as described below. herein. Rather, these embodiments are provided so that this The term "heterocycle' or "heterocyclic” refers to satu disclosure will be thorough and complete, and will fully rated or partially unsaturated monocyclic, bicyclic ortricyclic convey the scope of the invention to those skilled in the art. groups having from 3 to 15 atoms, in some instances 3 to 7. The terminology used in the description of the invention with at least one heteroatom in at least one of the rings, said 45 heteroatom being selected from O, S or N. Each ring of the herein is for the purpose of describing particular embodi heterocyclic group can contain one or two O atoms, one or ments only and is not intended to be limiting of the invention. two S atoms, one to four Natoms, provided that the total As used in the description of the invention and the appended number of heteroatoms in each ring is four or less and each claims, the singular forms “a”, “an and “the are intended to ring contains at least one carbon atom. The fused rings com include the plural forms as well, unless the context clearly 50 pleting the bicyclic or tricyclic heterocyclic groups may con indicates otherwise. Additionally, as used herein, the term tain only carbon atoms and may be saturated or partially “and/or includes any and all combinations of one or more of unsaturated. The N and Satoms may optionally be oxidized the associated listed items and may be abbreviated as “7”. and the Natoms may optionally be quaternized. Heterocyclic Unless otherwise defined, all technical and scientific terms also refers to alkyl groups containing said monocyclic, bicy used herein have the same meaning as commonly understood 55 clic or tricyclic heterocyclic groups. Examples of heterocy by one of ordinary skill in the art to which this invention clic rings include, but are not limited to, 2- or 3-piperidinyl, 2 belongs. or 3-piperazinyl, 2- or 3-morpholinyl. All such heterocyclic All publications, U.S. patent applications, U.S. patents and groups may also be optionally Substituted as described below other references cited herein are incorporated by reference in The term "heteroaryl refers to an aromatic group in a their entireties. 60 single or fused ring system having from 5 to 15 ring atoms, in The term “alkyl refers to straight or branched chain satu some instances 5 to 10, which have at least one heteroatom in rated or partially unsaturated hydrocarbon groups having at least one of the rings, said heteroatom being selected from from 1 to 20 carbon atoms, in some instances 1 to 8 carbon O. S or N. Each ring of the heteroaryl group can contain one atoms. The term “lower alkyl refers to alkyl groups contain or two O atoms, one or two S atoms, one to four Natoms, ing 1 to 6 carbonatoms. Examples of alkyl groups include, but 65 provided that the total number of heteroatoms in each ring is are not limited to, methyl, ethyl, isopropyl, tert-butyl, 3-hex four or less and each ring contains at least one carbon atom. enyl, and 2-butynyl. By “unsaturated' is meant the presence The fused rings completing the bicyclic or tricyclic groups US 7,491.695 B2 21 22 may contain only carbon atoms and may be saturated, par tuted alkyl, unsubstituted cycloalkyl, unsubstituted heterocy tially unsaturated or aromatic. In structures where the lone clic, unsubstituted aryl, unsubstituted heteroaryl, hydroxy, pair of electrons of a nitrogen atom is not involved in com alkoxy, aryloxy, acyl, amino, amido, carboxy, carboxyalkyl, pleting the aromatic pi electron system, the N atoms may carboxyaryl, mercapto, Sulfinyl, Sulfonyl, Sulfonamido, ami optionally be quaternized or oxidized to the N-oxide. Het dino, carbamoyl, guanidino orureido, and optionally contain eroaryl also refers to alkyl groups containing said cyclic ing one to three additional heteroatoms selected from O, S or groups. Examples of monocyclic heteroaryl groups include, N. but are not limited to pyrrolyl pyrazolyl pyrazolinyl, imida The term “carboxy' refers to the group —CO.H. Zolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiaz The term "carboxyalkyl” refers to the group —COR olyl, furanyl, thienyl, oxadiazolyl pyridyl, pyrazinyl, pyrim 10 wherein R is alkyl, cycloalkyl or heterocyclic. idinyl, pyridazinyl, and triazinyl. Examples of bicyclic The term “carboxyaryl” refers to the group —COR heteroaryl groups include, but are not limited to indolyl, wherein R, is aryl or heteroaryl. benzothiazolyl, benzoxazolyl, benzothienyl, quinolinyl, tet The term "cyano” refers to the group —CN. rahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, ben The term “formyl refers to the group —C(=O)H, also Zopyranyl, indolizinyl, benzofuranyl, isobenzofuranyl. 15 denoted CHO. chromonyl, coumarinyl, benzopyranyl, cinnolinyl, quinox The term “halo,” “halogen' or “halide” refers to fluoro, alinyl, indazolyl, purinyl, pyrrolopyridinyl, furopyridinyl, fluorine or fluoride, chloro, chlorine or chloride, bromo, bro thienopyridinyl, dihydroisoindolyl, and tetrahydroquinoli mine or bromide, and iodo, iodine or iodide, respectively. nyl. Examples of tricyclic heteroaryl groups include, but are The term “oxo” refers to the bivalent group —O, which is not limited to carbazolyl, benzindolyl, phenanthrollinyl, Substituted in place of two hydrogen atoms on the same acridinyl, phenanthridinyl, and Xanthenyl. All Such heteroaryl carbon to form a carbonyl group. groups may also be optionally substituted as described below. The term “mercapto” refers to the group—SR, wherein R, The term “hydroxy' refers to the group —OH. is hydrogen, alkyl, cycloalkyl, heterocyclic, aryl or het The term “alkoxy' refers to the group —OR, wherein R. eroaryl. is alkyl, cycloalkyl or heterocyclic. Examples include, but are 25 The term “nitro” refers to the group —NO. not limited to methoxy, ethoxy, tert-butoxy, cyclohexyloxy The term “trifluoromethyl refers to the group —CF. and tetrahydropyranyloxy. The term “sulfinyl" refers to the group S(=O)R. The term “aryloxy” refers to the group —OR, wherein R, wherein R is alkyl, cycloalkyl, heterocyclic, aryl or het is aryl or heteroaryl. Examples include, but are not limited to eroaryl. phenoxy, benzyloxy and 2-maphthyloxy. 30 The term "sulfonyl" refers to the group S(=O), R. The term “acyl refers to the group—C(=O)—R wherein wherein R is alkyl, cycloalkyl, heterocyclic, aryl or het R is alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl. eroaryl. Examples include, but are not limited to, acetyl, benzoyland The term "aminosulfonyl" refers to the group NR S furoyl. (=O), Rs wherein R2 is hydrogen, alkyl, cycloalkyl, het The term "amino acyl indicates an acyl group that is 35 erocyclic, aryl or heteroaryl; and R is alkyl, cycloalkyl, derived from an amino acid. heterocyclic, aryl or heteroaryl. The term “amino” refers to an —NRR group wherein R. The term “sulfonamido” refers to the group —S(=O)— and R are independently selected from the group consisting NRR wherein RandR are independently selected from the of hydrogen, alkyl, cycloalkyl, heterocyclic, aryl and het group consisting of hydrogen, alkyl, cycloalkyl, heterocyclic, eroaryl. Alternatively, R and R together form a heterocyclic 40 aryl or heteroaryl. Alternatively, R and R together form a ring of 3 to 8 members, optionally substituted with unsubsti heterocyclic ring of 3 to 8 members, optionally substituted tuted alkyl, unsubstituted cycloalkyl, unsubstituted heterocy with unsubstituted alkyl, unsubstituted cycloalkyl, unsubsti clic, unsubstituted aryl, unsubstituted heteroaryl, hydroxy, tuted heterocyclic, unsubstituted aryl, unsubstituted het alkoxy, aryloxy, acyl, amino, amido, carboxy, carboxyalkyl, eroaryl, hydroxy, alkoxy, aryloxy, acyl, amino, amido, car carboxyaryl, mercapto, Sulfinyl, Sulfonyl, Sulfonamido, ami 45 boxy, carboxyalkyl, carboxyaryl, mercapto, Sulfinyl, dino, carbamoyl, guanidino orureido, and optionally contain Sulfonyl, Sulfonamido, amidino, carbamoyl, guanidino or ing one to three additional heteroatoms selected from O, S or ureido, and optionally containing one to three additional het N. eroatoms selected from O, S or N. The term "amido” refers to the group C(=O)NRR, The term “carbamoyl refers to a group of the formula wherein RandR are independently selected from the group 50 —N(R)—C(=O)—OR, wherein R, is selected from hydro consisting of hydrogen, alkyl, cycloalkyl, heterocyclic, aryl gen, alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl; and R. and heteroaryl. Alternatively, R, and R together form a het is selected from alkyl, cycloalkyl, heterocylic, aryl or het erocyclic ring of 3 to 8 members, optionally substituted with eroaryl. unsubstituted alkyl, unsubstituted cycloalkyl, unsubstituted The term “guanidino” refers to a group of the formula heterocyclic, unsubstituted aryl, unsubstituted heteroaryl, 55 N(R) C(=NR) NR,R, wherein R,R,R, and Rare hydroxy, alkoxy, aryloxy, acyl, amino, amido, carboxy, car independently selected from hydrogen, alkyl, cycloalkyl, het boxyalkyl, carboxyaryl, mercapto, Sulfinyl, Sulfonyl, Sul erocyclic, arylorheteroaryl. Alternatively, RandR, together fonamido, amidino, carbamoyl, guanidino or ureido, and form a heterocyclic ring or 3 to 8 members, optionally sub optionally containing one to three additional heteroatoms stituted with unsubstituted alkyl, unsubstituted cycloalkyl, selected from O, S or N. 60 unsubstituted heterocyclic, unsubstituted aryl, unsubstituted The term "amidino” refers to the group C(=NR)NR,R, heteroaryl, hydroxy, alkoxy, aryloxy, acyl, amino, amido, wherein R, is selected from the group consisting of hydrogen, carboxy, carboxyalkyl, carboxyaryl, mercapto, Sulfinyl, Sul alkyl, cycloalkyl, heterocyclic, aryl and heteroaryl; and R, fonyl, Sulfonamido, amidino, carbamoyl, guanidino or ure and R, are independently selected from the group consisting ido, and optionally containing one to three additional heteroa of hydrogen, alkyl, cycloalkyl, heterocyclic, aryl and het 65 toms selected from O, S or N. eroaryl. Alternatively, R, and R, together form a heterocyclic The term “ureido” refers to a group of the formula ring of 3 to 8 members, optionally substituted with unsubsti —N(R)—C(=O)—NRR, wherein R, R and R, are US 7,491.695 B2 23 24 independently selected from hydrogen, alkyl, cycloalkyl, het A “stable compound or “stable structure” refers to a com erocyclic, aryl or heteroaryl. Alternatively, R, and R. pound that is sufficiently robust to survive isolation to a useful together form a heterocyclic ring of 3 to 8 members, option degree of purity and formulation into an efficacious therapeu ally substituted with unsubstituted alkyl, unsubstituted tic agent. cycloalkyl, unsubstituted heterocyclic, unsubstituted aryl, The term “amino acid refers to the common natural (ge unsubstituted heteroaryl, hydroxy, alkoxy, aryloxy, acyl, netically encoded) or synthetic amino acids and common amino, amido, carboxy, carboxyalkyl, carboxyaryl, mer derivatives thereof, known to those skilled in the art. When capto, Sulfinyl, Sulfonyl, Sulfonamido, amidino, carbamoyl, applied to amino acids, “standard” or “proteinogenic’ refers guanidino or ureido, and optionally containing one to three to the genetically encoded 20 amino acids in their natural 10 configuration. Similarly, when applied to amino acids, additional heteroatoms selected from O, S or N. "unnatural or “unusual refers to the wide selection of non The term “optionally substituted is intended to expressly natural, rare or synthetic amino acids such as those described indicate that the specified group is unsubstituted or Substi by Hunt, S. in Chemistry and Biochemistry of the Amino tuted by one or more suitable substituents, unless the optional Acids, Barrett, G. C., Ed., Chapman and Hall: New York, Substituents are expressly specified, in which case the term 15 1985. indicates that the group is unsubstituted or substituted with The term “residue” with reference to an amino acid or the specified Substituents. As defined above, various groups amino acid derivative refers to a group of the formula: may be unsubstituted or substituted (i.e., they are optionally Substituted) unless indicated otherwise herein (e.g., by indi cating that the specified group is unsubstituted). The term “substituted when used with the terms alkyl, cycloalkyl, heterocyclic, aryland heteroaryl refers to an alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl group having one N-teH X RAA or more of the hydrogen atoms of the group replaced by O substituents independently selected from unsubstituted alkyl, 25 unsubstituted cycloalkyl, unsubstituted heterocyclic, unsub wherein R is an amino acid side chain, and n=0, 1 or 2 in stituted aryl, unsubstituted heteroaryl, hydroxy, alkoxy, ary this instance. loxy, acyl, amino, amido, carboxy, carboxyalkyl, car The term “fragment” with respect to a dipeptide, tripeptide boxyaryl, halo, oxo, mercapto, Sulfinyl, Sulfonyl, 30 or higher order peptide derivative indicates a group that con Sulfonamido, amidino, carbamoyl, guanidino, ureido and tains two, three or more, respectively, amino acid residues. groups of the formulas —NRC(=O)R - NRC The term “amino acid side chain” refers to any side chain (=NR)R - OC(=O)NR,R,gigs OC(=O)R ii - OC from a standard or unnatural amino acid, and is denoted R. (=O)CR - NRSO.R., or - NRSONRR, For example, the side chain of alanine is methyl, the side wherein R. R. R. R. R. R. R. R. R. R. R. and 35 chain of valine is isopropyl and the side chain of tryptophanis R, are independently selected from hydrogen, unsubstituted 3-indolylmethyl. alkyl, unsubstituted cycloalkyl, unsubstituted heterocyclic, The term “agonist” refers to a compound that duplicates at unsubstituted aryl or unsubstituted heteroaryl; and wherein least Some of the effect of the endogenous ligand of a protein, R and R are independently selected from unsubstituted receptor, enzyme or the like. alkyl, unsubstituted cycloalkyl, unsubstituted heterocyclic, 40 The term “antagonist” refers to a compound that inhibits at unsubstituted aryl or unsubstituted heteroaryl. Alternatively, least Some of the effect of the endogenous ligand of a protein, R. and R. R., and R or R and R, together form a receptor, enzyme or the like. heterocyclic ring of 3 to 8 members, optionally substituted The term “growth hormone secretagogue' (GHS) refers to with unsubstituted alkyl, unsubstituted cycloalkyl, unsubsti any exogenously administered compound or agent that tuted heterocyclic, unsubstituted aryl, unsubstituted het 45 directly or indirectly stimulates or increases the endogenous eroaryl, hydroxy, alkoxy, aryloxy, acyl, amino, amido, car release of growth hormone, growth hormone-releasing hor boxy, carboxyalkyl, carboxyaryl, mercapto, Sulfinyl, mone, or Somatostatin in an animal, in particular, a human. A Sulfonyl, Sulfonamido, amidino, carbamoyl, guanidino or GHS may be peptidic or non-peptidic in nature, in some ureido, and optionally containing one to three additional het 50 instances, with an agent that can be administered orally. In eroatoms selected from O, S or N. In addition, the term Some instances, the agent can induce a pulsatile response. “substituted for aryl and heteroaryl groups includes as an The term “modulator” refers to a compound that imparts an option having one of the hydrogen atoms of the group effect on a biological or chemical process or mechanism. For replaced by cyano, nitro or trifluoromethyl. example, a modulator may increase, facilitate, upregulate, A substitution is made provided that any atoms normal 55 activate, inhibit, decrease, block, prevent, delay, desensitize, Valency is not exceeded and that the Substitution results in a deactivate, downregulate, or the like, a biological or chemical stable compound. Generally, when a substituted form of a process or mechanism. Accordingly, a modulator can be an group is present, such substituted group is preferably not “agonist' oran'antagonist.” Exemplary biological processes further substituted or, if substituted, the substituent comprises or mechanisms affected by a modulator include, but are not only a limited number of Substituted groups, in some 60 limited to, receptor binding and hormone release or secretion. instances 1, 2, 3 or 4. Such substituents. Exemplary chemical processes or mechanisms affected by a When any variable occurs more than one time in any con modulator include, but are not limited to, catalysis and stituent or in any formula herein, its definition on each occur hydrolysis. rence is independent of its definition at every other occur The term “variant' when applied to a receptor is meant to rence. Also, combinations of substituents and/or variables are 65 include dimers, trimers, tetramers, pentamers and other bio permissible only if Such combinations result in stable com logical complexes containing multiple components. These pounds. components can be the same or different. US 7,491.695 B2 25 26 The term "peptide' refers to a chemical compound com tives in Solid Phase Synthesis. Peptides, Polypeptides and prised of two or more amino acids covalently bonded Oligonucleotides; Epton, R., Ed., SPCC Ltd.: Birmingham, together. UK; p 205), polyacrylate (CLEARTM), polyacrylamide, poly The term "peptidomimetic' refers to a chemical compound urethane, PEGA polyethyleneglycol poly(N,N-dimethy designed to mimic a peptide, but which contains structural lacrylamide) co-polymer, Meldal, M. Tetrahedron Lett. 1992, differences through the addition or replacement of one of 33, 3077-3080), cellulose, etc. These materials can optionally more functional groups of the peptide in order to modulate its contain additional chemical agents to form cross-linked activity or other properties, such as Solubility, metabolic sta bonds to mechanically stabilize the structure, for example bility, oral bioavailability, lipophilicity, permeability, etc. polystyrene cross-linked with divinylbenezene (DVB, usu This can include replacement of the peptide bond, side chain 10 ally 0.1-5%, preferably 0.5-2%). This solid support can modifications, truncations, additions of functional groups, include as non-limiting examples aminomethyl polystyrene, etc. When the chemical structure is not derived from the hydroxymethyl polystyrene, benzhydrylamine polystyrene peptide, but mimics its activity, it is often referred to as a (BHA), methylbenzhydrylamine (MBHA) polystyrene, and “non-peptide peptidomimetic.” other polymeric backbones containing free chemical func The term "peptide bond refers to the amide —C(=O)— 15 tional groups, most typically, —NH2 or —OH, for further NH functionality with which individual amino acids are derivatization or reaction. The term is also meant to include typically covalently bonded to each other in a peptide. “Ultraresins” with a high proportion (“loading) of these The term “protecting group' refers to any chemical com functional groups such as those prepared from polyethylene pound that may be used to prevent a potentially reactive imines and cross-linking molecules (Barth, M.; Rademann, J. functional group. Such as an amine, a hydroxyl or a carboxyl, J. Comb. Chem. 2004, 6,340-349). At the conclusion of the on a molecule from undergoing a chemical reaction while synthesis, resins are typically discarded, although they have chemical change occurs elsewhere in the molecule. A number been shown to be able to be reused such as in Frechet, J. M. J.; of such protecting groups are known to those skilled in the art Haque, K. E. Tetrahedron Lett. 1975, 16, 3055. and examples can be found in “Protective Groups in Organic In general, the materials used as resins are insoluble poly Synthesis. Theodora W. Greene and Peter G. Wuts, editors, 25 mers, but certain polymers have differential solubility John Wiley & Sons, New York, 3' edition, 1999 ISBN depending on Solvent and can also be employed for Solid 0471160199. Examples of amino protecting groups include, phase chemistry. For example, polyethylene glycol can be but are not limited to, phthalimido, trichloroacetyl, benzy utilized in this manner since it is soluble in many organic loxycarbonyl, tert-butoxycarbonyl, and adamantyloxycarbo Solvents in which chemical reactions can be conducted, but it nyl. In some embodiments, amino protecting groups are car 30 is insoluble in others, such as diethyl ether. Hence, reactions bamate amino protecting groups, which are defined as an can be conducted homogeneously in Solution, then the prod amino protecting group that when bound to an amino group uct on the polymer precipitated through the addition of forms a carbamate. In other embodiments, amino carbamate diethyl ether and processed as a solid. This has been termed protecting groups are allyloxycarbonyl (Alloc), benzyloxy “liquid-phase' chemistry. carbonyl (Cbz), 9-fluorenylmethoxycarbonyl (Fmoc), tert 35 butoxycarbonyl (Boc) and O.C.-dimethyl-3,5-dimethoxyben The term “linker' when used in reference to solid phase Zyloxycarbonyl (Ddz). For a recent discussion of newer chemistry refers to a chemical group that is bonded covalently nitrogen protecting groups: Theodoridis, G. Tetrahedron to a solid Support and is attached between the Support and the 2000, 56, 2339-2358. Examples of hydroxyl protecting Substrate typically in order to permit the release (cleavage) of groups include, but are not limited to, acetyl, tert-butyldim 40 the substrate from the solid support. However, it can also be ethylsilyl (TBDMS), trityl (Trt), tert-butyl, and tetrahydropy used to impart stability to the bond to the solid support or ranyl (THP). Examples of carboxyl protecting groups merely as a spacer element. Many solid Supports are available include, but are not limited to methyl ester, tert-butyl ester, commercially with linkers already attached. benzyl ester, trimethylsilylethyl ester, and 2.2.2-trichloroet Abbreviations used for amino acids and designation of hyl ester. 45 peptides follow the rules of the IUPAC-IUB Commission of The term “solid phase chemistry” refers to the conduct of Biochemical Nomenclature in J. Biol. Chem. 1972, 247,977 chemical reactions where one component of the reaction is 983. This document has been updated: Biochem. J., 1984, covalently bonded to a polymeric material (Solid Support as 219, 345-373 Eur: J. Biochem., 1984, 138, 9-37; 1985, 152, defined below). Reaction methods for performing chemistry 1 Internat. J. Pept. Prot. Res., 1984, 24, following p 84, J. on Solid phase have become more widely known and estab 50 Biol. Chem., 1985, 260, 14-42; Pure Appl. Chem., 1984, 56, lished outside the traditional fields of peptide and oligonucle 595-624. Amino Acids and Peptides, 1985, 16,387-410; and otide chemistry. in Biochemical Nomenclature and Related Documents, 2nd The term "solid support,”99 “solid&g phase' or “resin refers to edition, Portland Press, 1992, pp. 39-67. Extensions to the a mechanically and chemically stable polymeric matrix uti rules were published in the JCBN/NC-IUB Newsletter 1985, lized to conduct solid phase chemistry. This is denoted by 55 1986, 1989; see Biochemical Nomenclature and Related “Resin,” “P-” or the following symbol: Documents, 2nd edition, Portland Press, 1992, pp 68-69. The term “effective amount’ or “effective' is intended to designate a dose that causes a relief of symptoms of a disease or disorder as noted through clinical testing and evaluation, 60 patient observation, and/or the like, and/or a dose that causes a detectable change in biological or chemical activity. The detectable changes may be detected and/or further quantified Examples of appropriate polymer materials include, but by one skilled in the art for the relevant mechanism or pro are not limited to, polystyrene, polyethylene, polyethylene cess. As is generally understood in the art, the dosage will glycol, polyethylene glycol grafted or covalently bonded to 65 vary depending on the administration routes, symptoms and polystyrene (also termed PEG-polystyrene, TentaGelTM, body weight of the patient but also depending upon the com Rapp, W.; Zhang, L.; Bayer, E. In Innovations and Persepc pound being administered. US 7,491.695 B2 27 28 Administration of two or more compounds in combina tion” means that the two compounds are administered closely -continued enough in time that the presence of one alters the biological effects of the other. The two compounds can be administered simultaneously (concurrently) or sequentially. Simultaneous 5 administration can be carried out by mixing the compounds prior to administration, or by administering the compounds at the same point in time but at different anatomic sites or using different routes of administration. The phrases “concurrent administration', 'administration in combination', 'simulta 10 neous administration” or “administered simultaneously as used herein, means that the compounds are administered at the same point in time or immediately following one another. In the latter case, the two compounds are administered at 15 times sufficiently close that the results observed are indistin guishable from those achieved when the compounds are administered at the same point in time. The term “pharmaceutically active metabolite' is intended to mean a pharmacologically active product produced through metabolism in the body of a specified compound. The term “solvate” is intended to mean a pharmaceutically acceptable solvate form of a specified compound that retains the biological effectiveness of such compound. Examples of Solvates, without limitation, include compounds of the inven 25 tion in combination with water, isopropanol, ethanol, metha nol, DMSO, ethyl acetate, acetic acid, or ethanolamine. 1. Compounds Novel macrocyclic compounds of the present invention 30 include macrocyclic compounds comprising a building block structure including a tether component that undergoes cyclization to form the macrocyclic compound. The building wherein (Z) is the site of a covalent bond of T to Z, and Z. block structure can comprise amino acids (standard and 35 unnatural), hydroxy acids, hydrazino acids, aza-amino acids, is as defined below for formula I, and wherein (X) is the site specialized moieties such as those that play a role in the of a covalent bond of T to X, and X is as defined below for introduction of peptide Surrogates and isosteres, and a tether formula I; L7 is —CH2— or —O—; U is —CRoRo or component as described herein. The tether component can be —C(=O)—, Roo is lower alkyl, Ro and Ro are each selected from the following: independently hydrogen, lower alkyl or substituted lower 40 alkyl; XX is 2 or 3; yy is 1 or 2: ZZ is 1 or 2; and aaa is 0 or 1. Macrocyclic compounds of the present invention further include those of formula I, formula II and/or formula III:
45 (I) R4 HOC (CH2) O O Y Y
50
(CH2)m PN. "S-ch) R RE X\ S T-Z. 55 or an optical isomer, enantiomer, diastereomer, racemate or stereochemical mixture thereof, wherein: 60 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-, 65 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 7,491.695 B2 29 30 R is hydrogen or the side chain of an amino acid, or m, n and pare each independently 0, 1 or 2; alternatively RandR togetherform a 4-, 5-, 6-, 7- or 8-mem T is a bivalent radical of formula IV: bered ring, optionally comprising an O. S or Natom in the ring, wherein the ring is optionally Substituted with Rs as defined below; or alternatively RandR together form a 3-, 5 wherein dande are each independently 0, 1, 2, 3, 4 or 5:Y 4-, 5-, 6- or 7-membered ring, optionally comprising an O.S and Zare each optionally present, U is —CRR - or or additional Natom in the ring, wherein the ring is optionally —C(=O)—and is bonded to X of formula I; W. Yand substituted with Rs as defined below: Zare each independently selected from the group con R is hydrogen or the side chain of an amino acid, or alternatively RandR togetherform a 3-, 4-, 5-, 6- or 7-mem 10 bered ring, optionally comprising an O or Satom in the ring, wherein the ring is optionally substituted with Rs as defined —CRRs , —CH=CH- with the configuration Z below, or alternatively, R and R7 or R and R togetherform or E. —C=C and the ring structures below: a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, optionally 15 comprising an O.S or additional Natom in the ring, wherein the ring is optionally substituted with R as defined below: R is hydrogen or the side chain of an amino acid, or 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 Ra and R, 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: Rs and R are each independently hydrogen or the side 25 chain of an amino 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: R is hydrogen, lower alkyl, substituted lower alkyl, 30 cycloalkyl, Substituted cycloalkyl, a heterocyclic group, or a substituted heterocyclic group, or alternatively R and R, 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 35 with Rs as described below: wherein G and G2 are each independently a covalent Rs is substituted for one or more hydrogenatoms on the 3-, bond or a bivalent radical selected from the group 4-, 5-, 6-, 7- or 8-membered-ring structure and is indepen dently selected from the group consisting of alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic 40 group, a Substituted heterocyclic group, aryl, Substituted aryl, heteroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, —CH=CH with the configuration Z or E, and oxo, amino, halogen, formyl, acyl, carboxy, carboxyalkyl, —C=C-, with G being bonded closest to the group carboxyaryl, amido, carbamoyl, guanidino, ureido, amidino, U; wherein any carbonatom in the rings not otherwise mercapto, Sulfinyl, Sulfonyl and Sulfonamido, or, alterna 45 defined, is optionally replaced by N, with the proviso tively, Rs is a fused cycloalkyl, a Substituted fused cycloalkyl, that the ring cannot contain more than four Natoms; a fused heterocyclic, a substituted fused heterocyclic, a fused K. K. K. K. and Ks are each independently O, aryl, a substituted fused aryl, a fused heteroaryl or a substi NR or S, wherein R is as defined below: tuted fused heteroaryl ring when substituted for hydrogen R and R are each independently hydrogen, lower atoms on two adjacent atoms; 50 alkyl, or substituted lower alkyl, or alternatively R. X is O. NR or N(R): and R together form a 3- to 12-membered cyclic wherein R is hydrogen, lower alkyl, substituted lower ring optionally comprising one or more heteroatoms alkyl, Sulfonyl, Sulfonamido or amidino and Rio is selected from the group consisting of O, S and N, hydrogen, lower alkyl, or substituted lower alkyl, or wherein the ring is optionally substituted with Rs as alternatively Ro and R together form a 3-, 4-, 5-, 6- or 55 defined above; 7-membered ring, optionally comprising an O. S or R. R. and Ra are each independently hydrogen, additional Natom in the ring, wherein the ring is option alkyl, substituted alkyl, cycloalkyl, substituted ally substituted with Rs as defined above: cycloalkyl, heterocyclic, Substituted heterocyclic, Z is O or NR, aryl, substituted aryl, heteroaryl, substituted het wherein R is hydrogen, lower alkyl, or substituted lower 60 eroaryl, formyl, acyl, carboxyalkyl, carboxyaryl, alkyl, or alternatively R and R or R and R together amido, amidino, Sulfonyl or Sulfonamido; form a 4-, 5-, 6-, 7- or 8-membered heterocyclic ring, Ra and Rs are each independently hydrogen, lower optionally comprising an O. S or additional Natom in alkyl, Substituted lower alkyl, R, wherein R is a the ring, wherein the ring is optionally Substituted with side chain of an amino acid Such as a standard or Rs as defined above: 65 unusual amino acid, or alternatively Ra and Rs Z is O or NR2, wherein R is hydrogen, lower alkyl, or togetherform a 3- to 12-membered cyclic ring option substituted lower alkyl: ally comprising one or more heteroatoms selected US 7,491.695 B2 31 32 from the group consisting of O, S and N, or alterna with the proviso that T is not an amino acid residue, dipep tively one of R or Rs is hydroxy, alkoxy, aryloxy, tide fragment, tripeptide fragment or higher order pep amino, mercapto, carbamoyl, amidino, ureido or tide fragment comprising standard amino acids; guanidino while the other is hydrogen, lower alkyl or substituted lower alkyl, except when the carbon to 5 which R- and Rs are bonded is also bonded to another heteroatom; R2, R1, Ras and Ras are each optionally present and, when present, are substituted for one or more hydro 10 gen atoms on the indicated ring and each is indepen dently selected from the group consisting of halogen, trifluoromethyl, alkyl, substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substi tuted heterocyclic group, aryl, Substituted aryl, het 15 or an optical isomer, enantiomer, diastereomer, racemate or eroaryl, Substituted heteroaryl, hydroxy, alkoxy, ary stereochemical mixture thereof, loxy, amino, formyl, acyl, carboxy, carboxyalkyl, wherein: carboxyaryl, amido, carbamoyl, guanidino, ureido, Rso is —(CH2)CH, —CH(CH)(CH2)CH, amidino, cyano, nitro, mercapto, Sulfinyl, Sulfonyl —(CH2)CH(CH), —C(CH), —(CHRss), Rs, or and Sulfonamido; —CH(ORs)CH, whereinss is 1, 2 or 3; tt is 1 or 2: uu is 0, 1 or 2; and VV is 0, 1, 2, 3 or 4: Rss is hydrogen or C-C alkyl; R27 is optionally present and is substituted for one or Rs is amino, hydroxy, alkoxy, cycloalkyl or substituted more hydrogen atoms on the indicated ring and each cycloalkyl, and Rs7 is hydrogen, alkyl, acyl, amino acyl, is independently selected from the group consisting of 25 Sulfonyl, carboxyalkyl or carboxyaryl; alkyl, substituted alkyl, cycloalkyl, substituted Rs is hydrogen, C-C alkyl or C-C alkyl Substituted cycloalkyl, a heterocyclic group, a Substituted hetero with hydroxy or alkoxy; cyclic group, aryl, Substituted aryl, heteroaryl, Substi Rs is —(CHRs).Rs, wherein ww is 0, 1, 2 or 3: Rss is tuted heteroaryl, hydroxy, alkoxy, aryloxy, OXo, hydrogen, C-C alkyl, amino, hydroxy oralkoxy; Rso is aryl, amino, formyl, acyl, carboxy, carboxyalkyl, car 30 substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl or substituted cycloalkyl: boxyaryl, amido, carbamoyl, guanidino, ureido, ami Rs is hydrogen or C-C alkyl: dino, mercapto, Sulfinyl, Sulfonyl and Sulfonamido; X is O. NR or N(R): R2s, R29. Rso Rs.2, Ras Rs. Rs6 and Rs 7 are each wherein R is hydrogen, lower alkyl, substituted lower optionally present and, when no double bond is 35 alkyl, Sulfonyl, sulfonamido or amidino and R is present to the carbonatom to which it is bonded in the hydrogen, lower alkyl, or substituted lower alkyl: ring, two groups are optionally present, and when Zs is O or NR, wherein R is hydrogen, lower alkyl, or present, is Substituted for one hydrogen present in the substituted lower alkyl; and ring, or when no double bond is present to the carbon T is a bivalent radical of formula V: atom to which it is bonded in the ring, is substituted 40 - U -(CH2) W. Y-Z-(CH2) - (V) for one or both of the two hydrogen atoms present on wherein dande are independently 0, 1, 2, 3, 4 or 5;Y, and the ring and each is independently selected from the Z, are each optionally present; U is —CRR— or group consisting of alkyl, Substituted alkyl, —C(=O)—and is bonded to X of formula II, wherein cycloalkyl, Substituted cycloalkyl, a heterocyclic 45 Reo and R are each independently hydrogen, lower group, a Substituted heterocyclic group, aryl, Substi alkyl, or substituted lower alkyl, or alternatively R and tuted aryl, heteroaryl, substituted heteroaryl, R together form a 3- to 12-membered cyclic ring hydroxy, alkoxy, aryloxy, Oxo, amino, formyl, acyl, optionally comprising one or more heteroatoms selected carboxy, carboxyalkyl, carboxyaryl, amido, carbam from the group consisting of O, S and N, wherein the oyl, guanidino, ureido, amidino, mercapto, Sulfinyl, 50 ring is optionally substituted with Rs as defined above: sulfonyl, sulfonamido and, only if a double bond is WY, and Z are each independently selected from the present to the carbon atom to which it is bonded, group consisting of —O— —NR— —S-, halogen; and SO , SO , C(=O)—O , C(=O)—, Rao and Ra are each independently hydrogen, lower 55 C(=O) NH , NH-C(=O) , SO, alkyl, substituted lower alkyl, R as defined above, NH-, -NH SO , —CRR , —CH=CH or alternatively Rao and R, together form a 3- to with the configuration Z or E. —C=C , and the ring 12-membered cyclic ring optionally comprising one structures depicted below: or more heteroatoms selected from the group consist 60 ing of O, S and N wherein the ring is optionally substituted with Rs as defined above, or alternatively 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 65 lower alkyl, except when the carbon to which Rao and Rare bonded is also bonded to another heteroatom; US 7,491.695 B2 34 bamoyl, guanidino, ureido, amidino, mercapto, -continued sulfinyl, sulfonyl or sulfonamido: CH2ff CH2ff K is O or S; and G G G2 ff is 1, 2, 3, 4 or 5: with the proviso that T is not an amino acid residue, dipeptide fragment, tripeptide fragment or higher order wherein G and G2 are as defined above, and wherein peptide fragment comprising standard amino acids; or any carbon atom in the ring is optionally replaced by N, with the proviso that the aromatic ring cannot contain more than four Natoms and the cycloalkyl 10 ring cannot contain more than two Natoms; R is hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Sub stituted heterocyclic group, aryl, Substituted aryl, heteroaryl, substituted heteroaryl, formyl, acyl, 15 carboxyalkyl, carboxyaryl, amido, amidino, Sulfo nyl or Sulfonamido; Re and Rea are each independently hydrogen, lower alkyl, Substituted lower alkyl or R, or alterna tively R and R together form a 3- to 12-mem bered cyclic ring optionally comprising one or or an optical isomer, enantiomer, diastereomer, racemate or more heteroatoms selected from the group consist stereochemical mixture thereof, wherein: ing of O, S and N: or alternatively one of R and Ro is hydrogen, C-C alkyl or alternatively Ro and R, together form a 4-, 5-, 6-, 7- or 8-membered ring, optionally Ra is hydroxy, alkoxy, aryloxy, amino, mercapto, 25 carbamoyl, amidino, ureido or guanidino, while the comprising an O, N or Satom in the ring, wherein the ring is other is hydrogen, lower alkyl or substituted lower optionally substituted with Rs, as defined below: alkyl, except when the carbon to which Re and Rea R7 is, hydrogen, —(CH2)CH, —CH(CH)(CH), are bonded is also bonded to another heteroatom; CH, -(CH2)CH(CH), —(CH) R, or—CH(OR7) and R indicates the side chain of a standard or 30 CH or, alternatively R7 and Rio together form a 4-, 5-, 6-, 7 unusual amino acid; or 8-membered ring, optionally comprising an O. NorSatom Res and Rs are each optionally present, and, when in the ring, wherein the ring is optionally substituted with Rs. present are substituted for one or more hydrogen as defined below; wherein aa is 0, 1, 2, 3, 4 or 5: bb is 1, 2 or atoms on the ring and each is independently halo 3; cc is 0, 1, 2 or 3; and dd is 0, 1, 2, 3 or 4: R-7 is aryl, gen, trifluoromethyl, alkyl, Substituted alkyl, 35 substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl cycloalkyl, Substituted cycloalkyl, a heterocyclic or substituted cycloalkyl, Rz, is hydrogen, alkyl, acyl, amino group, a Substituted heterocyclic group, aryl. Sub acyl, Sulfonyl, carboxyalkyl or carboxyaryl; stituted aryl, heteroaryl, substituted heteroaryl, R7 is C-C alkyl, or alternatively R7 and R together hydroxy, alkoxy, aryloxy, amino, formyl, acyl, car form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris boxy, carboxyalkyl, carboxyaryl, amido, carbam 40 ing an O or Satom in the ring, wherein the ring is optionally oyl, guanidino, ureido, amidino, cyano, nitro, mer substituted with Rs, as defined below: capto, Sulfinyl, Sulfonyl or Sulfonamido; R is hydrogen, or alternatively R7 and R, togetherform Re and R, are each optionally present, and when no a 3-, 4-, 5-, 6- or 7-membered ring, optionally comprising an double bond is present to the carbonatom to which O, S or N atom in the ring, wherein the ring is optionally it is bonded in the ring, two groups are optionally 45 substituted with Rs, as defined below: present, and, when present, each is Substituted for Ra is hydrogen or C-C alkyl or alternatively Ra and R-7s one hydrogen present in the ring, or when no together form a 3-, 4-, 5-, 6- or 7-membered ring, optionally double bond is present to the carbonatom to which comprising an O, N or Satom in the ring, wherein the ring is it is bonded in the ring, is substituted for one or both optionally substituted with Rs as defined below: of the two hydrogen atoms present on the ring and 50 R7s is —(CHR-7s)R 79 or alternatively R-7s and R together each is independently alkyl, Substituted alkyl, form a 3-, 4-, 5-, 6- or 7-membered ring, optionally compris cycloalkyl, Substituted cycloalkyl, heterocyclic, ing an O, N or Satom in the ring, wherein the ring is option substituted heterocyclic, aryl, substituted aryl, het ally substituted with Rs, as defined below; wherein Rs is eroaryl, Substituted heteroaryl, hydroxy, alkoxy, hydrogen, C-C alkyl, amino, hydroxy or alkoxy, and Rio is aryloxy, oxo, amino, formyl, acyl, carboxy, car 55 boxyalkyl, carboxyaryl, amido, carbamoyl, guani selected from the group consisting of the following struc dino, ureido, amidino, mercapto, Sulfinyl, Sulfonyl, tures: sulfonamido and, only ifa double bond is present to the carbon atom to which it is bonded, halogen; Ro is optionally present, and when present is substi 60 tuted for one or more hydrogen atoms on the ring and each is independently alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, aryl. Sub stituted aryl, heteroaryl, substituted heteroaryl, 65 hydroxy, alkoxy, aryloxy, oxo, amino, formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, car US 7,491.695 B2 35 36
-continued -continued
10 wherein E. E. E. E. and Es are each optionally present and when present are each independently selected from the group consisting of halogen, trifluoromethyl, alkyl, Substituted alkyl, cycloalkyl, Substituted cycloalkyl, a heterocyclic group, a Substituted heterocyclic group, 15 aryl, substituted aryl, heteroaryl, substituted heteroaryl, hydroxy, alkoxy, aryloxy, cyano, Sulfinyl, Sulfonyl and Sulfonamido, and represent Substitution at one or more available positions on the monocyclic or bicyclic aro matic ring, wherein said Substitution is made with the same or different selected group member, and J and J are each independently O or S; Rs. Rs, and Rs are each independently substituted for one or more hydrogen atoms on the 3-, 4-, 5-, 6-, 7- or 8-mem 25 bered ring structure and are independently selected from the group consisting of alkyl, Substituted alkyl, cycloalkyl, Sub stituted cycloalkyl, a heterocyclic group, a Substituted hetero cyclic group, aryl, Substituted aryl, heteroaryl, Substituted heteroaryl, hydroxy, alkoxy, aryloxy, Oxo, amino, halogen, 30 formyl, acyl, carboxy, carboxyalkyl, carboxyaryl, amido, car bamoyl, guanidino, ureido, amidino, mercapto, Sulfinyl, Sul fonyl and Sulfonamido, or, alternatively, Rs. Rs, and Rs are each independently a fused cycloalkyl, a Substituted fused 35 cycloalkyl, a fused heterocyclic, a substituted fused hetero cyclic, a fused aryl, a Substituted fused aryl, a fused heteroaryl or a substituted fused heteroaryl ring when substituted for hydrogen atoms on two adjacent atoms; X is O, NR or N(R)"; wherein R is hydrogen, lower alkyl, substituted lower alkyl, Sulfonyl, Sulfonamido or amidino and Rio is hydrogen, lower alkyl, or substituted lower alkyl: Zo is O or NR, wherein R is hydrogen, lower alkyl, or 45 substituted lower alkyl; and T is the same as defined for T with the exception that U. is bonded to X of formula III. In some embodiments of the present invention, the com 50 pound can have one of the following structures:
2. O 55 N HN
NH HN 60
65 US 7,491.695 B2 37 38
-continued -continued Y.2. O
OMe s S v s NH 10
15
C
NH 25
30
35 O N) {( s F 40
NH HN
S 45
50
2. O C
55
60
65 US 7,491.695 B2 39 40
-continued -continued 2. O 2. O CF
( 10 ( -O O S
15
C
2. O 2O 2. O
NH K HNy D-\NH ").HN
S 30
F 35 C 2 9 2 O S. F O S C s N HN N HN 40 \ \ O O NH HN NH HN
y-O 45 O
50 F 2. O 2. O O e S. C O - S. C N HN- 55 N HN \ )—o \ )- NH NH
( O 60 ( -O
65 US 7,491.695 B2 41 42
-continued -continued C O O NH HN
NH O ( 10 NH HN -Q 8 -
15
C 2 O 2 O 2O O ( S. C NH HN s N HN )- O \ )—o NH HN 8.NH HN 25 8 - 30 Cl, O C O 35 NH HN 2. O O O S. F NH HN N HN 40 O NH HN S- 45
or an optical isomer, enantiomer, diastereomer, racemate or stereochemical mixture thereof. 50 C The present invention includes isolated compounds. An isolated compound refers to a compound that, in some embodiements, comprises at least 10%, at least 25%, at least 2. O 50% or at least 70% of the compounds of a mixture. In some embodiments, the compound, pharmaceutically acceptable O e S C 55 salt thereof or pharmaceutical composition containing the N HN compound exhibits a statistically significant binding and/or antagonist activity when tested in biological assays at the \ )=o human ghrelin receptor. NH HN In the case of compounds, salts, or Solvates that are solids, ( 60 it is understood by those skilled in the art that the inventive compounds, salts, and Solvates may exist in different crystal -O or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas. 65 The compounds of formula I, II and/or III disclosed herein have asymmetric centers. The inventive compounds may exist as single Stereoisomers, racemates, and/or mixtures of US 7,491.695 B2 43 44 enantiomers and/or diastereomers. All Such single stereoiso with the defined substituent R. mers, racemates, and mixtures thereof are intended to be The use of the following symbol indicates a single bond or within the scope of the present invention. In particular an optional double bond: it embodiments, however, the inventive compounds are used in Embodiments of the present invention further provide optically pure form. The terms “S” and “R” configuration as 5 intermediate compounds formed through the synthetic meth used herein are as defined by the IUPAC 1974 Recommen ods described herein to provide the compounds of formula I, dations for Section E, Fundamentals of Stereochemistry II and/or III. The intermediate compounds may possess util (Pure Appl. Chem. 1976, 45, 13-30.) tity as a therapeutic agent for the range of indications Unless otherwise depicted to be a specific orientation, the described herein and/or a reagent for further synthesis meth present invention accounts for all Stereoisomeric forms. The 10 ods and reactions. compounds may be prepared as a single stereoisomer or a mixture of Stereoisomers. The non-racemic forms may be 2. Synthetic Methods obtained by either synthesis or resolution. The compounds The compounds of formula I, II and/or II can be synthe may, for example, be resolved into the component enanti sized using traditional solution synthesis techniques or solid omers by Standard techniques, for example formation of dias 15 phase chemistry methods. In either, the construction involves tereomeric pairs via salt formation. The compounds also may four phases: first, synthesis of the building blocks comprising be resolved by covalently bonding to a chiral moiety. The recognition elements for the biological target receptor, plus diastereomers can then be resolved by chromatographic sepa one tether moiety, primarily for control and definition of ration and/or crystallographic separation. In the case of a conformation. These building blocks are assembled together, chiral auxiliary moiety, it can then be removed. As an alter typically in a sequential fashion, in a second phase employing standard chemical transformations. The precursors from the native, the compounds can be resolved through the use of assembly are then cyclized in the third stage to provide the chiral chromatography. Enzymatic methods of resolution macrocyclic structures. Finally, the post-cyclization process could also be used in certain cases. ing fourth stage involving removal of protecting groups and As generally understood by those skilled in the art, an 25 optional purification provides the desired final compounds. “optically pure' compound is one that contains only a single Synthetic methods for this general type of macrocyclic struc enantiomer. As used herein, the term “optically active' is intended to mean a compound comprising at least a sufficient ture are described in Intl. Pat. Appls. WO 01/25257, WO excess of one enantiomer over the other such that the mixture 2004/111077, WO 2005/012331 and WO 2005/012332, rotates plane polarized light. Optically active compounds including purification procedures described in WO 2004/ have the ability to rotate the plane of polarized light. The 30 111077 and WO 2005/O12331. excess of one enantiomer over another is typically expressed In some embodiments of the present invention, the macro as enantiomeric excess (e.e.). In describing an optically active cyclic compounds of formula I, II and/or III may be synthe compound, the prefixes D and L or Rand S are used to denote sized using solid phase chemistry on a soluble or insoluble the absolute configuration of the molecule about its chiral polymer matrix as previously defined. For Solid phase chem center(s). The prefixes “d” and “1” or (+) and (-) are used to 35 istry, a preliminary stage involving the attachment of the first denote the optical rotation of the compound (i.e., the direction building block, also termed “loading.” to the resin must be in which a plane of polarized light is rotated by the optically performed. The resin utilized for the present invention pref active compound). The “1” or (-) prefix indicates that the erentially has attached to it a linker moiety, L. These linkers compound is levorotatory (i.e., rotates the plane of polarized are attached to an appropriate free chemical functionality, light to the left or counterclockwise) while the “d' or (+) 40 usually an alcohol oramine, although others are also possible, prefix means that the compound is dextrarotatory (i.e., rotates on the base resin through standard reaction methods known in the plane of polarized light to the right or clockwise). The sign the art, Such as any of the large number of reaction conditions of optical rotation, (-) and (+), is not related to the absolute developed for the formation of ester or amide bonds. Some configuration of the molecule, R and S. linker moieties for the present invention are designed to allow 45 for simultaneous cleavage from the resin with formation of A compound of the invention having the desired pharma the macrocycle in a process generally termed “cyclization cological properties will be optically active and, can be com release.” (van Maarseveen, J. H. Solid phase synthesis of prised of at least 90% (80% e.e.), at least 95% (90% e.e.), at heterocycles by cyclization/cleavage methodologies. Comb. least 97.5% (95% e.e.) or at least 99% (98% e.e.) of a single Chem. High Throughput Screen. 1998, 1, 185-214; Ian W. isomer. 50 James, Linkers for Solid phase organic synthesis. Tetrahedron Likewise, many geometric isomers of double bonds and 1999, 55, 4855-4946; Eggenweiler, H.-M. Linkers for solid the like can also be present in the compounds disclosed phase synthesis of Small molecules: coupling and cleavage herein, and all such stable isomers are included within the techniques. Drug Discovery Today 1998, 3,552-560; Backes, present invention unless otherwise specified. Also included in B.J.; Ellman, J. A. Solid support linker strategies. Curr: Opin. the invention are tautomers and rotamers of formula I, II 55 Chem. Biol. 1997, 1,86-93. Of particular utility in this regard and/or III. for compounds of the invention is the 3-thiopropionic acid The use of the following symbols at the right refers to linker. (Hojo, H.; Aimoto, S. Bull. Chem. Soc. Jpn. 1991, 64. Substitution of one or more hydrogen atoms of the indicated 111-117; Zhang, L.; Tam, J. J. Am. Chem. Soc. 1999, 121, ring 3311-3320.) 60 Such a process provides material of higher purity as only cyclic products are released from the solid Support and mini mal contamination with the linear precursor occurs as would S/S× (O, S, NH happen in solution phase. After sequential assembly of all the building blocks and tether into the linear precursor using C2 XMYR 65 known or standard reaction chemistry, base-mediated intramolecular attack on the carbonyl attached to this linker by an appropriate nucleophilic functionality that is part of the US 7,491.695 B2 45 46 tether building block results information of the amide orester KGaA, Darmstadt, Germany), PepTech (Burlington, Mass., bond that completes the cyclic structure as shown (Scheme 1). USA), Synthetech (Albany, Oreg., USA) or synthesized An analogous methodology adapted to Solution phase can through standard methodologies known to those in the art. also be applied as would likely be preferable for larger scale Ddz-amino acids were either obtained commercially from applications. 5 Orpegen (Heidelberg, Germany) or Advanced ChemTech (Louisville, Ky., USA) or synthesized using standard meth ods utilizing Ddz-OPh or Ddz-N. (Birr, C.: Lochinger, W.; Scheme 1. Cyclization-release Strategy Stahnke, G.; Lang, P. The O.C.-dimethyl-3,5-dimethoxyben Zyloxycarbonyl (DdZ) residue, an N-protecting group labile 10 toward weak acids and irradiation. Justus Liebigs Ann. Chem. 1972, 763, 162-172.) Bts-amino acids were synthesized by known methods. (Vedes, E., Lin, S.; Klapara, A.; Wang, J. Cyclization-release Linker J BB-BB-BB Base “Heteroarene-2-sulfonyl Chlorides (BtsCl. ThsC1): Reagents --- for Nitrogen Protection and >99% Racemization-Free Phe HY-Tether ) (Y = O, NH) 15 nylglycine Activation with SOCl. 'J. Am. Chem. Soc. 1996, O 118, 9796-9797. Also WO 01/25257, WO 2004/111077) N-Alkylamino acids, in particular N-methylamino acids, are commercially available from multiple vendors (Bachem, --- Novabiochem, Advanced ChemTech, ChemImpex). In addi tion, N-alkylamino acid derivatives were accessed via litera ture methods. (Hansen, D. W., Jr.; Pilipauskas, D. J. Org. Chem. 1985, 50,945-950.) - B. Tethers 25 Tethers were obtained from the methods previously Although this description accurately represents the path described in Intl. Pat. Appl. WO 01/25257, WO 2004/111077, way for one of the methods of the present invention, the WO 2005/012331 and U.S. Provisional Patent Application thioester Strategy, another method of the present invention, Ser. No. 60/622,055. Procedures for synthesis of tethers as that of ring-closing metathesis (RCM), proceeds through a described herein are presented in the Examples below. Exem modified route where the tether component is actually 30 plary tethers (T) include, but are not limited to, the following: assembled during the cyclization step. However, in the RCM methodology as well, assembly of the building blocks pro ceeds sequentially, followed by cyclization (and release from the resin if solid phase). An additional post-cyclization pro cessing step is required to remove particular byproducts of the 35 RCM reaction, but the remaining Subsequent processing is done in the same manner as for the thioester or analogous base-mediated cyclization strategy. Moreover, it will be understood that steps including the methods provided herein may be performed independently or 40 at least two steps may be combined. Additionally, steps including the methods provided herein, when performed independently or combined, may be performed at the same temperature or at different temperatures without departing from the teachings of the present invention. 45 Novel macrocyclic compounds of the present invention include those formed by a novel process including cyclization ofa building block structure to form a macrocyclic compound comprising a tether component described herein. Accord ingly, the present invention provides methods of manufactur 50 ing the compounds of the present invention comprising (a) assembling building block structures, (b) chemically trans forming the building block structures, (c) cyclizing the build ing block structures including a tether component, (d) remov ing protecting groups from the building block structures, and 55
(e) optionally purifiying the product obtained from step (d). In some embodiments, assembly of the building block structures may be sequential. In further embodiments, the synthesis ZZ (Z) methods are carried out using traditional Solution synthesis techniques or Solid phase chemistry techniques. 60 A. Amino Acids Amino acids, Boc- and Fmoc-protected amino acids and side chain protected derivatives, including those of N-methyl and unnatural amino acids, were obtained from commercial suppliers for example Advanced ChemTech (Louisville, Ky., 65 USA), Bachem (Bubendorf, Switzerland), ChemImpex (Wood Dale, Ill., USA), Novabiochem (subsidiary of Merck US 7,491.695 B2 47 48
-continued -continued
U (Z2) \-o A. ) 333. Resin wash 2 XDCM, 2 X (DCM-MeOH), 2 X DCM (all for 5 min) and 3:7 DIPEA-DCM (for 3 min); dried quickly used (X) (Z2) or immediately Yu 2 10 HOat (2eq), DMF (2.5 mL, degassed) 5%. AcOHDMF (2.5 mL, C S 50° C., 16 h degassed 50-70° C., 16 h Macrocycle Macrocycle (Zs) 15 For a representative macrocycle with tether T1, AA=Leu, AA-Leu, AA=Phe, the application of the HOAt method U O shown in Scheme 2 provided the cyclic peptidomimetic in 10% yield, while the acetic acid method was more effective, and gave 24% overall yield of the same macrocycle. This latter methodology was particularly effective for compounds containing His(Mts) residues. For example, with tether T8, AA=Phe, AAAcp, AA, =His(Mts), the macrocycle was and intermediates in the manufacture thereof, wherein (Z) is 25 obtained in 20% overall yield, although the majority of the the site of a covalent bond of T to Z, Zs or Zo and Z, Zs and product no longer had the Mts group on histidine (15:1 versus Zo are defined above for formulae I, II and III, respectively, still protected). and wherein (X) is the site of a covalent bond of T to X, X, or Synthesis of representative macrocyclic compounds of the X and X, X and X are defined above for formula I, II and III, present invention are shown in the Examples below. Table 1A respectively, L, is —CH2— or —O—; U, is —CRoRo 30 below presents a Summary of the synthesis of 224 represen or —C(=O)—: Roo is lower alkyl; Ro, and Ro are each tative compounds of the present invention. The reaction meth independently hydrogen, lower alkyl or substituted lower odology employed for the construction of the macrocyclic alkyl; XX is 2 or 3; yy is 1 or 2: ZZ is 1 or 2; and aaa is 0 or 1. molecule is indicated in Column 2 and relates to the particular C. Solid Phase Techniques scheme of the synthetic strategy, for example, use of the Specific solid phase techniques for the synthesis of the 35 thioester strategy as shown in FIG. 2 or the RCM approach as shown in FIG. 3. Column 3 indicates if any substituents are macrocyclic compounds of the invention have been described present on N. Columns 4-6 and 8 indicate the individual in WO 01/25257, WO 2004/111077, WO 2005/012331 and building blocks employed for each compound, amino acids, WO 2005/012332. Solution phase synthesis routes, including hydroxy acids or tether utilizing eitherstandard nomenclature methods amenable to larger scale manufacture, were 40 or referring to the building block designations presented else described in U.S. Provisional Patent Application Ser. Nos. where in this application. Column 7 indicates the method 60/622,055 and 60/642,271. used for attachment of the tether, either a Mitsunobu reaction In certain cases, however, the lability of protecting groups (previously described in WO 01/25257) or reductive amina precluded the use of the standard basic medium for cycliza tion (previously described in WO 2004/111077). The relevant tion in the thioester strategy discussed above. In these cases, 45 deprotection and coupling protocols as appropriate for the either of two acidic methods was employed to provide mac nature of the building block are performed utilizing standard rocyclization under acid conditions. One method utilized procedures and those described in WO 2004/111077 for the HOAc, while the other method employed HOAt (Scheme 2). assembly of the cyclization precursors. The building blocks For example, the acetic acid cyclization was used for com are listed in the opposite order from which they are added in pound 219. 50 order to correlate the building block number with standard After executing the deprotection of the Ddz or Boc group peptide nomenclature. Hence BB is added first, followed by on the tether, the resin was washed sequentially with DCM BB, then BB, finally the tether(T). In the case of the RCM, (2x), DCM-MeOH (1:1, 2x), DCM (2x), and DIPEA-DCM the tether is not formed completely until the cyclization step, (3:7, 1x). The resin was dried under vacuum for 10 min, then but the portion of the tether attached to BB is still added at added immediately to a solution of HOAc in degassed DMF 55 this stage of the sequence, unless it is already part of that (5% v/v). The reaction mixture was agitated at 50-70°C. O/N. building block. The final macrocycles are obtained after The resin was filtered, washed with THF, and the combined application of the appropriate deprotection sequences. If any filtrate and washes evaporated under reduced pressure (water reaction was required to be carried out post-cyclization, it is aspirator, then oil pump) to afford the macrocycle. listed in Column 9. All of the macrocycles presented in Table 60 1A were purified and met internal acceptance criteria. Yields (Column 10) are either isolated or as calculated based upon Scheme 2: CLND analysis. It should be noted that compounds 58 and 99 Alternative CY clization Methodologies were not cyclized and represent the linear analogues of com pounds 10 and 133, respectively. The lack of binding potency % FA% ES 65 He observed with these linear analogues illustrates the impor DCM, rt, 1 h tance of the macrocyclic structural feature for the desired activity.
US 7,491.695 B2 55
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention Tether Attachment Additional Yield Compound Method Tether Reaction** (%)*
1 Mitsunobu Reaction Boc-T9 None 10.1 2 Mitsunobu Reaction Boc-T9 None 13.8 3 Mitsunobu Reaction Boc-T9 None 10.3 4 Mitsunobu Reaction Boc-T9 None 4.6 5 Mitsunobu Reaction Boc-T9 None 8.6 6 Mitsunobu Reaction Ddz-T9 None 8.1 7 Mitsunobu Reaction Ddz-T9 None 8.8 8 Mitsunobu Reaction Boc-T8 None 20.9 9 Mitsunobu Reaction Boc-T9 None 9.7 10 Mitsunobu Reaction Boc-T9 None 9.9 11 Mitsunobu Reaction Boc-T8 None 9.9 12 Mitsunobu Reaction Boc-T8 None 2.9 13 Mitsunobu Reaction Boc-T8 None 5.8 14 Mitsunobu Reaction Boc-T8 None 27.5 15 Mitsunobu Reaction Boc-T9 None 19.5 16 Mitsunobu Reaction Boc-T9 None 23.9 17 Reductive Amination Boc-T9 None 24.8 Reaction 18 Mitsunobu Reaction Boc-T8 None 6.8 19 Mitsunobu Reaction Boc-T8 None 12.7 2O Mitsunobu Reaction Boc-T8 None 22.0 21 Mitsunobu Reaction Boc-T8 None 24.7 22 Mitsunobu Reaction Boc-T8 None 10.3 23 Mitsunobu Reaction Boc-T9 None 32.6 24 Mitsunobu Reaction Boc-T9 None 22.4 25 Mitsunobu Reaction Boc-T9 None 21.0 26 Mitsunobu Reaction Boc-T9 None 15.5 27 Mitsunobu Reaction Boc-T9 None 2O2 28 Mitsunobu Reaction Boc-T9 None 31.6 29 Mitsunobu Reaction Boc-T9 None 26.1 30 Mitsunobu Reaction Boc-T9 None 31.9 31 Mitsunobu Reaction Boc-T9 None 21.9 Reaction 32 Reductive Amination Boc-T9 None 6.7 33 Mitsunobu Reaction Boc-T9 None 7.5 34 Mitsunobu Reaction Boc-T9 None 4.2 35 Mitsunobu Reaction Boc-T33a None 9.4 36 Mitsunobu Reaction Boc-T33b None 3.0 37 Mitsunobu Reaction T41 + TBA None 24.6 38 Mitsunobu Reaction T42 + Tel Hydrogenation 44.2 39 Mitsunobu Reaction Boc-T8 None 21.4 40 Mitsunobu Reaction Boc-T8 None 8.6 41 Mitsunobu Reaction Boc-T9 None O6 42 Mitsunobu Reaction Boc-T9 None 1.7 43 Mitsunobu Reaction Boc-T9 None 0.4 44 Mitsunobu Reaction Boc-T1 None 7.8 45 Mitsunobu Reaction Ddz-T8 None 1.6 46 Mitsunobu Reaction Boc-T8 None 3.6 47 Mitsunobu Reaction Boc-T8 None 9.2 48 Mitsunobu Reaction Boc-T8 None 7.5 49 Reductive Amination Boc-T9 None 7.5 Reaction 50 Mitsunobu Reaction Ddz-T9 None O.1 51 Mitsunobu Reaction Boc-T9 None 6.6 52 Mitsunobu Reaction Boc-T9 None 8.7 53 Mitsunobu Reaction Ddz-T9 None 8.3 S4 Mitsunobu Reaction Ddz-T9 None 6.2 55 Mitsunobu Reaction Boc-T9 None 8.0 56 Mitsunobu Reaction Boc-T9 None 9.3 57 Mitsunobu Reaction Boc-T9 None 8.9 58 Mitsunobu Reaction Boc-T9 No cyclization 5.9 59 Mitsunobu Reaction Boc-T9 None 8.0 60 Mitsunobu Reaction Boc-T9 None 13.1 61 Mitsunobu Reaction Boc-T9 None 8.4 62 Mitsunobu Reaction Boc-T9 None 7.0 63 Mitsunobu Reaction Boc-T9 None 11.7 64 Mitsunobu Reaction Boc-T9 None 8.5 65 Mitsunobu Reaction Boc-T9 None 8.6 66 Mitsunobu Reaction Boc-T9 None 15.8 67 Mitsunobu Reaction Boc-T9 None 11.7 68 Mitsunobu Reaction Ddz-T9 None 7.9 69 Mitsunobu Reaction Ddz-T9 None 11.2 70 Mitsunobu Reaction Ddz-T9 None 1O.O 71 Mitsunobu Reaction Ddz-T9 None 9.9 US 7,491.695 B2 57 58
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention 72 Mitsunobu Reaction Boc-T9 None 5.2 73 Mitsunobu Reaction Boc-T9 None 6.8 74 Mitsunobu Reaction Boc-T9 None 6.O 75 Mitsunobu Reaction Boc-T9 None 9.5 76 Mitsunobu Reaction Boc-T9 None 15.1 77 Mitsunobu Reaction Boc-T12 None 12.6 78 Mitsunobu Reaction Boc-T27 None 6.8 79 Mitsunobu Reaction Boc-T9 None 1.9 8O Mitsunobu Reaction Boc-T9 None 1.3 81 Mitsunobu Reaction Boc-T1 None 5.3 82 Mitsunobu Reaction Boc-T3 None 3.9 83 Mitsunobu Reaction Boc-T16 None 1.8 84 Mitsunobu Reaction Boc-T4 None 2.6 85 Mitsunobu Reaction Boc-TS None 4.7 86 Mitsunobu Reaction Boc-T14 None 0.4 87 Mitsunobu Reaction Boc-T9 None 4.8 88 Mitsunobu Reaction Ddz-T9 None 18.8 89 Mitsunobu Reaction Ddz-T9 None 16.5 90 Mitsunobu Reaction Boc-T9 None 8.5 91 Mitsunobu Reaction Ddz-T9 None 6.8 92 Mitsunobu Reaction Ddz-T9 None 9.1 93 Mitsunobu Reaction Boc-T9 None 9.2 94 Mitsunobu Reaction Boc-T9 None 21.8 95 Mitsunobu Reaction Boc-T9 None 9.3 96 Mitsunobu Reaction Boc-T9 None 7.0 97 Mitsunobu Reaction Boc-T9 None 9.2 98 Mitsunobu Reaction Boc-T9 None 5.3 99 Mitsunobu Reaction Boc-T9 No cyclization 0.4 OO Mitsunobu Reaction Boc-T9 None 0.4 O1 Mitsunobu Reaction Boc-T9 None 9.0 O2 Mitsunobu Reaction Boc-T9 None 5.8 O3 Mitsunobu Reaction Boc-T9 None 2.9 O4 Mitsunobu Reaction Boc-T9 None 9.3 05 Mitsunobu Reaction Boc-T9 None 1.9 O6 Mitsunobu Reaction Boc-T3 None 6.3 O7 Mitsunobu Reaction Boc-TS None 4.2 O8 Mitsunobu Reaction Boc-T12 None 8.3 09 Mitsunobu Reaction Boc-T11 None O.1 10 Mitsunobu Reaction Boc-T9 None 2.9 11 Mitsunobu Reaction Boc-T9 None 3.0 12 Mitsunobu Reaction Boc-T9 None 3.2 13 Mitsunobu Reaction Boc-T9 None 6.9 14 Mitsunobu Reaction Boc-T16 None 2.9 15 Mitsunobu Reaction Boc-T6 None O.S 16 Mitsunobu Reaction Ddz-T8 None 1.8 17 Mitsunobu Reaction Boc-T9 None 9.7 18 Mitsunobu Reaction Boc-T9 None 19 Mitsunobu Reaction Boc-T9 None 2.2 2O Reductive Amination Boc-T9 None 7.5 Reaction 21 Mitsunobu Reaction Boc-T9 None 5.8 22 Mitsunobu Reaction Boc-T8 None 23 Mitsunobu Reaction Boc-T8 None 3.6 24 Mitsunobu Reaction Boc-T8 None 9.8 25 Mitsunobu Reaction Boc-T8 None 5.8 26 Mitsunobu Reaction Boc-T8 None 9.8 27 Mitsunobu Reaction Boc-T8 None 4.5 28 Mitsunobu Reaction Boc-T8 None 7.8 29 Mitsunobu Reaction Boc-T8 None 1.O 30 Mitsunobu Reaction Boc-T8 None 8.8 31 Mitsunobu Reaction Boc-T8 None S.O 32 Reductive Amination Boc-T8 None 7.0 Reaction 33 Reductive Amination Boc-T8 None 9.5 Reaction 34 Mitsunobu Reaction Boc-T8 None 2.0 35 Mitsunobu Reaction Boc-T8 None 4.0 36 Mitsunobu Reaction Boc-T8 None 3.3 37 Mitsunobu Reaction Boc-T8 None 9.0 38 Mitsunobu Reaction Boc-T8 None 3.8 39 Reductive Amination Boc-T8 None 8.4 Reaction 40 Reductive Amination Boc-T8 None 6.7 Reaction 41 Reductive Amination Boc-T8 None 5.7 Reaction US 7,491.695 B2 59 60
TABLE 1 A-continued Synthesis of Representative Compounds o the Present Invention 42 Reductive Amination Boc-T8 None 17.0 Reaction 43 Reductive Amination Boc-T8 None 16.1 Reaction 44 Reductive Amination Boc-T9 None 5.7 Reaction 45 Reductive Amination Boc-T9 None 4.9 Reaction 46 Reductive Amination Boc-T9 None 23.3 Reaction 47 Reductive Amination Boc-T9 None 14.4 Reaction 48 Reductive Amination Boc-T9 None 25.4 Reaction 49 Reductive Amination Boc-T9 None 11.4 Reaction 50 Reductive Amination Boc-T9 None 7.0 Reaction 51 Mitsunobu Reaction Ddz-T9 None 52 Reductive Amination Ddz-T9 None Reaction 53 Mitsunobu Reaction Boc-T8 None S4 Mitsunobu Reaction Boc-T8 None 55 Mitsunobu Reaction Boc-T8 None 56 Mitsunobu Reaction Boc-T8 None 57 Mitsunobu Reaction Boc-T8 None 58 Mitsunobu Reaction Boc-T8 None 59 Mitsunobu Reaction Boc-T8 None 60 Reductive Amination Boc-T8 None Reaction 61 Reductive Amination Boc-T8 None Reaction 62 Reductive Amination Boc-T8 None 6.7 Reaction 63 Mitsunobu Reaction Boc-T8 None O.O 64 Mitsunobu Reaction Boc-T8 None 2.5 65 Mitsunobu Reaction Boc-T8 None 3.0 66 Mitsunobu Reaction Boc-T8 None 1.1 67 Mitsunobu Reaction Boc-T8 None 5.3 68 Mitsunobu Reaction Boc-T8 None 4.2 69 Mitsunobu Reaction Boc-T9 None 7.0 70 Reductive Amination Boc-T9 None 4.5 Reaction 71 Reductive Amination Boc-T9 None 6.4 Reaction 72 Mitsunobu Reaction Boc-T9 None 2.0 73 Mitsunobu Reaction Boc-T9 None 6.8 74 Mitsunobu Reaction Boc-T9 None 3.9 75 Mitsunobu Reaction Boc-T9 None S.1 76 Mitsunobu Reaction Boc-T9 None 9.4 77 Mitsunobu Reaction Boc-T11 None 9.3 78 Mitsunobu Reaction Boc-T28 None 1.2 79 Mitsunobu Reaction Boc-T29 None 8.6 8O Mitsunobu Reaction Boc-T30 None O.O 81 Mitsunobu Reaction T41 + Te7 None 49.5 82 Mitsunobu Reaction T41 +TB7 Hydrogenation 47.7 83 Mitsunobu Reaction T42 +TB7 None 59.0 84 Mitsunobu Reaction T42 +TB7 Hydrogenation SO.6 85 Mitsunobu Reaction T41 + TB6 None 12.4 86 Mitsunobu Reaction T.42 + TB6 None 3.0 87 Mitsunobu Reaction T41 +TB3 None 30.9 88 Mitsunobu Reaction T42 + TB3 None 34.9 89 Mitsunobu Reaction T42 +TB3 Hydrogenation 24.0 90 Mitsunobu Reaction T41 + TBA Hydrogenation 32.5 91 Mitsunobu Reaction T42 + TBA None 32.2 92 Mitsunobu Reaction T.42 + TBA Hydrogenation 22.2 93 Mitsunobu Reaction T41 +TB1 None 47.7 94 Mitsunobu Reaction T41 + Tel Hydrogenation 23.7 95 Mitsunobu Reaction T42 +TB1 None 66.8 96 Mitsunobu Reaction None 13.0 97 Mitsunobu Reaction None 10.6 99 Reductive Amination None 16.0 Reaction 200 Mitsunobu Reaction None 14.7 2O1 Reductive Amination Reductive 32.4 Reaction amination US 7,491.695 B2 61 62
TABLE 1 A-continued Synthesis of Representative Compounds of the Present Invention reaction with formaldehyde 2O2 Reductive Amination Boc-T9 Acetylation 14.2 Reaction 2O3 Reductive Amination Boc-T8 Reductive 7.7 Reaction amination reaction with formaldehyde 204 Reductive Amination Boc-T8 Acetylation 11.5 Reaction 205 Mitsunobu Reaction Boc-T9 None 19.9 2O6 Mitsunobu Reaction Boc-T34 None 26.2 2O7 Mitsunobu Reaction Boc-T9 None 3. 208 Mitsunobu Reaction Boc-T8 None 16.7 209 Mitsunobu Reaction Boc-T9 None 8.6 210 Reductive Amination Boc-T8 None .1 Reaction 211 Reductive Amination Boc-T8 None 3. Reaction 212 Mitsunobu Reaction Boc-T9 None .2 213 Reductive Amination Boc-T9 None O Reaction 214 Mitsunobu Reaction T41 +TB3 Hydrogenation 4.9 215 216 Reductive Amination Boc-T9 None .6 Reaction 218 Mitsunobu Reaction Boc-T8 None O.1 219 Reductive Amination Boc-T8 None 9.0 Reaction 220 Mitsunobu Reaction Boc-T9 None S.O 221 Mitsunobu Reaction Boc-T9 None 4.9 222 Mitsunobu Reaction Boc-T9 None 1.7 223 Mitsunobu Reaction Boc-T9 None 20.4 224 Mitsunobu Reaction T41 + TB2 Hydrogenation 8.2 225 Reductive Amination Boc-T9 None O.O Reaction 226 Reductive Amination Boc-T9 None 3.5 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 Table 1B below presents a summary of the synthesis of 122 a ment of the tether. The building blocks are listed in the oppo representative compounds of the present invention, and Table site order from which they are added in order to correlate the 1C presents the synthesis of an additional 15 representative building block number with standard peptide nomenclature. compounds. For Table 1B, the reaction methodology employed for the construction of the macrocyclic molecule is Column 8 indicates if any additional reaction chemistry was indicated in Column 2 and relates to the particular scheme of as applied, such as to remove auxiliary protection or to reduce a the synthetic strategy. Columns 3-6 indicate the individual double bond (as was performed with many RCM intermediate building blocks employed for each compound, amino acids or products). All of the macrocycles in Tables 1B and 1C were tether utilizing either standard nomenclature or referring to purified and met the acceptance criteria. Yields (Column the building block designations presented elsewhere in this 9-10) are either isolated or as calculated based upon CLND application. Column 7 indicates the method used for attach analysis.
TABLE 1B Synthesis of Representative Compounds of the Present Invention Macrocyclic Compound Assembly Method BB BB, BBs Tether 298 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T33a 299 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T9 301 Thioester Strategy BtS-Tyr(But) Boc-Acp Boc-Phe(3-CI) Ddz-T8 303 Thioester Strategy Bts-Val Boc-(4O)Acp Boc-Phe Boc-T8 305 Thioester Strategy Bts-Ile Boc-(D)NMeAla Boc-(D)His(Mts) Boc-T9 306 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T11 307 RCM Strategy Fmoc-Cpg Fmoc-(D)NMeAla Fmoc-(D)Phe(4-F) T.42 + TB6 3O8 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T8 309 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T9 310 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)3-Thi Boc-T9 311 Thioester Strategy Boc-Cpg Boc-(D)NMeAla Boc-(D)Tyr(3-tBu) Boc-T9
US 7,491.695 B2 65 66
TABLE 1 B-continued Synthesis of Representative Compounds of the Present Invention 389 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T68 390 Thioester Strategy Bts-Ile Boc-Acp Boc-Phe(3-CI) Boc-T18 391 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(3,4,5-triF) Boc-T9 392 Thioester Strategy Bts-Ile Boc-Acp Boc-Phe(3-CI) Boc-T40 393 Thioester Strategy Bts-Ile Boc-Acp Boc-Phe(3-CI) Boc-T45 394 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T38 395 RCM Strategy Fmoc-Ile Fmoc-(4N)Acp Fmoc-Phe(3-CI) T41 + TB2 396 Thioester Strategy BtS-Acp Boc-(D)NMeAla Boc-Phe(3-CI) Boc-T8 397 Thioester Strategy BtS-Acp NMeAla Boc-Phe(3-CI) Boc-T8 398 RCM Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) T.42 + TB6 399 Thioester Strategy Bts-Ile Boc-Acp Boc-Phe(3-CI) Boc-T33b 400 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T66 4O1 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F) Boc-T8 4O2 Thioester Strategy Bts-Ile Boc-Acp Boc-Phe(3-CI) Boc-T8 403 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe Boc-T33a 40S Thioester Strategy Bts-Nva Boc-(D)NMeAla Boc-(D)Phe(4-F Boc-T33a 4O6 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-F Boc-T75a 407 Thioester Strategy Bts-Ile Boc-(D)NMeAla Boc-(D)Phe(4-F Boc-T33a 4.08 Thioester Strategy Bts-Ile Boc-(D)NMeAla Boc-(D)Phe(4-F Boc-T75a 409 Thioester Strategy Bts-Val Boc-(D)NMeAla Boc-(D)Phe(4-F Boc-T33a 410 RCM Strategy BtS-Nwa Boc-(D)NMeAla Boc-(D)Phe Boc-T75a 415 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T33a 417 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T69 430 Thioester Strategy Bts-Cpg Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T75a 431 Thioester Strategy Bts-Ile Boc-(D)NMeAla Boc-(D)Phe Boc-T33a 432 Thioester Strategy Bts-Ile Boc-(D)NMeAla Boc-(D)Phe(4-CI) Boc-T33a Additional Compound Tether Attachment Reaction** Amount (mg)* Yield (%)*
298 Mitsunobu Reaction None 29.7 2 299 Mitsunobu Reaction None 54.1 7 301 Mitsunobu Reaction None 36.5 O 303 Mitsunobu Reaction None 60 6 305 Reductive Amination None 110 31 Reaction 306 Mitsunobu Reaction None 51 8 307 Mitsunobu Reaction None 13.6 O 3O8 Mitsunobu Reaction None 43.8 4 309 Mitsunobu Reaction None 38.2 3 310 Mitsunobu Reaction None 33.3 1 311 Reductive Amination None 18.6 S.1 Reaction 312 Mitsunobu Reaction None 42.9 4 313 Mitsunobu Reaction None 38.2 3 314 Mitsunobu Reaction None 39.7 2 315 Mitsunobu Reaction None 35.3 1 316 Mitsunobu Reaction None 40.7 3 317 Mitsunobu Reaction None 37.6 2 318 Mitsunobu Reaction None 36.1 1 319 Mitsunobu Reaction None 37.5 1 32O Mitsunobu Reaction None 43.4 2 321 Mitsunobu Reaction None 34.5 1 322 Mitsunobu Reaction None 40.8 2 323 Mitsunobu Reaction None 27.3 8 324 Mitsunobu Reaction None 38.6 2 325 Mitsunobu Reaction None 33.7 O 326 Mitsunobu Reaction None 37.5 O 327 Mitsunobu Reaction None 35.2 1 328 Mitsunobu Reaction None 31.5 O 329 Mitsunobu Reaction None 26.9 8 330 Mitsunobu Reaction None 38.4 1 331 Mitsunobu Reaction None 37 1 332 Mitsunobu Reaction None 30.6 9 333 Mitsunobu Reaction None 49.6 8 334 Mitsunobu Reaction None 32 1 335 Reductive Amination None 62.2 8 Reaction 336 Mitsunobu Reaction None 37.7 2 337 Reductive Amination Hydrogenolysis 67.5 7 Reaction 338 Mitsunobu Reaction None 60 2O 339 Mitsunobu Reaction None 63 2O 340 Reductive Amination None 14.4 4 Reaction 341 Mitsunobu Reaction None 48 14 342 Mitsunobu Reaction None 37.7 10 US 7,491.695 B2 67 68
TABLE 1 B-continued Synthesis of Representative Compounds of the Present Invention
343 SO Ol Reac ion None 91.3 2 344 SO Ol Reac ion None 22.1 345 SO Ol Reac ion None 48 346 SO Ol Reac ion None S2.1 347 SO Ol Reac ion None 17.1 348 SO Ol Reac ion None 104.4 3 349 SO Ol Reac ion None 23.6 350 SO Ol Reac ion None 44 351 SO Ol Reac ion None 39.1 352 SO Ol Reac ion None 15.7 353 SO Ol Reac ion None 47.8 2 3S4 SO Ol Reac ion None 26.8 355 SO Ol Reac ion None 36.8 356 SO Ol Reac ion None 10 357 SO Ol Reac ion None 4.1.8 358 SO Ol Reac ion None 26.1 359 SO Ol Reac ion None 43.6 360 SO Ol Reac ion None 36.3 361 SO Ol Reac ion None 36.3 362 SO Ol Reac ion Hydrogenation 59.4 363 SO Ol Reac ion Hydrogenation 4.1.8 364 SO Ol Reac ion Hydrogenation 49.1 365 SO Ol Reac ion Hydrogenation 31.2 366 SO Ol Reac ion Hydrogenation 33.3 367 SO Ol Reac ion None 21.1 368 SO Ol Reac ion None 21.8 369 SO Ol Reac ion None 21.1 370 SO Ol Reac ion Hydrogenation 8.9 371 SO Ol Reac ion Hydrogenation 9.9 372 SO Ol Reac ion None 30.9 373 SO Ol Reac ion None 34.9 374 SO Ol Reac ion None 42.7 375 SO Ol Reac ion None 22.3 376 SO Ol Reac ion None 7.5 377 SO Ol Reac ion None 14.6 378 SO Ol Reac ion None 65.3 379 SO Ol Reac ion None 36.3 380 SO Ol Reac ion None 16.5 381 SO Ol Reac ion None 22.5 382 SO Ol Reac ion None 24.5 383 SO Ol Reac ion None 25.2 384 SO Ol Reac ion None 21.9 385 SO Ol Reac ion None 23.3 386 SO Ol Reac ion None 12 387 SO Ol Reac ion None 17.1 388 SO Ol Reac ion None 30 389 SO Ol Reac ion None 16.1 390 SO Ol Reac ion None 28.7 391 SO Ol Reac ion None 45.4 392 SO Ol Reac ion None 4.3 393 SO Ol Reac ion None 2.1 394 SO Ol Reac ion None 3.7 395 SO Ol Reac ion Hydrogenation O.2 396 SO Ol Reac ion None 2.3 397 SO Ol Reac ion None 1.4 398 SO Ol Reac ion Hydrogenation 3.8 399 SO Ol Reac ion None 5.7 400 SO Ol Reac ion None 28.3 4O1 SO Ol Reac ion None 31.5 4O2 SO Ol Reac ion None 29.1 403 SO Ol Reac ion None 103 40S SO Ol Reac ion None 38.8 4O6 SO Ol Reac ion None 45 407 SO Ol Reac ion None 138.5 4.08 SO Ol Reac ion None 146.2 409 SO Ol Reac ion None 125.7 410 SO Ol Reac ion None 36 415 SO Ol Reac ion None 127.5 417 SO Ol Reac ion None 45.6 430 SO Ol Reac ion None 50.7 431 SO Ol Reac ion None 57.9 432 SO Ol Reac ion None 141
*Overall Yield: based on theoretical resin loading, starting Ol ~500 mg resin **Additional reactions conducted post-cyclization to reach the esired product
US 7,491.695 B2 71 72
TABLE 2A-continued TABLE 2A-continued Characterization for Representative pounds of the Present Invention MW Calc MS (M + H)+ (g/mol) Found 550.7 551 550.7 551 10 550.7 551 564.8 565 564.8 565 564.8 565 564.8 565 524.7 525 15 538.7 539 S81.1 581 S64.7 565 560.7 S61 576.7 577 622.8 623 622.8 623 596.8 597 547.7 S48 547.7 S48 547.7 S48 553.7 554 552.7 553 25 484.6 485 512.7 513 1.O 2 510.7 511 536.7 537 523.7 524 30 523.7 524 460.6 461 474.6 475 474.6 475 488.7 489 488.7 489 35 523.7 524 508.7 509 506.7 507 520.7 521 456.6 457 458.6 459 470.6 471 40 472.7 473 458.6 459 472.6 473 SO4.7 505 518.7 519 520.7 521 45 520.7 521 532.7 533 534.7 535 SO4.7 505 506.7 507 518.7 519 50 S64.7 565 566.7 567 SO4.6 505 532.7 533 522.7 523 550.7 551 55 560.7 S61 588.7 589 460.6 461 582.7 583 549.7 550 S46.7 547 488.7 489 60 581.7 582 547.7 S48 536.7 537 552.7 553 506.7 507 S80.8 581 65 SO6.6 507 558.7 559
US 7,491.695 B2 75 76 2.5 min gradient to 70% ACN, 30% of a solution 50 mM of TABLE 2B-continued CHCOONH in H.O. 3. Isocratic plateau of 10 min at 70% ACN, 30% of a solution Analytical Characterization for Representative 50 mM of CHCOONH in H.O. Compounds of the Present Invention 4.5 min gradient to 40% ACN, 60% of a solution 50 mM of Molecular MW Calc MS (M + H)" CHCOONH in H.O. Compound Formula (g/mol) Found 5. Isocratic plateau of 10 min at 40% ACN, 60% of a solution 431 C31H44N4O4 536.7 537 50 mM of CHCOONH in H.O. 432 C31H43N4O4C 571.2 571 6. Flow: 0.5 mL/min 10 7. Column temperature: room temperature Notes 8. Sample temperature: room temperature 1. Molecular formulas and molecular weights are calculated automatically from the structure via ActivityBase software (IDBS, Guildford, Surrey, UK). 2. M+H obtained from LC-MS analysis using standard methods. Method Chiral C: Grad 55A-05 (column: Chiralcel OD-RH, 3. All analyses conducted on material after preparative purification by the 0.46 cmx15 cm): methods described above. 1.40 min isocratic 55%/45% of ACN/50 mM CHCOONH 15 in HO 2.5 min gradient to 70%/30% of ACN/50 mMCHCOONH TABLE 2C in HO Analytical Characterization for Representative 3. 10 min isocratic 70%/30% of ACN/50 mM CHCOONH Compounds of the Present Invention in HO 4.5 min gradient to 55%/44% of ACN/50 mMCHCOONH Molecular MW Calc MS (M + H)" in HO Compound Formula (g/mol) Found 5. 10 min isocratic 55%/45% of ACN/50 mM CHCOONH in HO 6. Flow: 0.5 mL/min 25 7. Column temperature: room temperature 439 C32B39N4OSC 595.1 595 8. Sample temperature: room temperature Method Chiral D: Grad Isol 00B 05 (column: Chiralcel OD 442 C26H38N4OS 486.6 487 RH, 0.46 cmx15 cm): 30 1.40 min isocratic 27%/73% of ACN/50 mM CHCOONH in HO 446 C24H3SN4OSF 478.6 479 2.5 min gradient to 70%/30% of ACN/50 mMCHCOONH in HO 3. 10 min isocratic 70%/30% of ACN/50 mM CHCOONH 449 C31H42NSO4C 584.1 S84 35 in HO Notes 4.5 min gradient to 27%/73% of ACN/50 mMCHCOONH 1. Molecular formulas and molecular weights are calculated automatically in HO from the structure via ActivityBase software (IDBS, Guildford, Surrey, UK). 5 2. M+H obtained from LC-MS analysis using standard methods. 10 min isocratic 27%/73% of ACN/50 mM CHCOONH 3. All analyses conducted on material after preparative purification by the in HO methods described above. 40 Flow: 0.5 mL/min . Column temperature: room temperature D. Chiral Purity Determination . Sample temperature: room temperature General methods for the HPLC determination of stereoi Someric purity were employed according to techniques . Biological Methods The compounds of the present invention were evaluated for known to those skilled in the art and further optimized for the 45 their ability to interact at the human ghrelin receptor utilizing compounds of the present invention. a competitive radioligand binding assay, fluorescence assay Method Chiral A: Grad35A-05 (column: Chiralcel AS-RH or Aequorin functional assay as described below. Such meth 0.46 cmx15 cm): ods can be conducted in a high throughput manner to permit 1. Isocratic plateau of 40 min at 35% ACN, 65% of a 50 mM the simultaneous evaluation of many compounds. solution of CHCOONH in H.O. 50 Specific assay methods for the human (GHS-R1a), Swine 2.5 min gradient to 70% ACN, 30% of a 50 mM solution of and rat GHS-receptors (U.S. Pat. No. 6.242,199, Intl. Pat. CHCOONH in H.O. Appl. Nos. WO 97/21730 and 97/22004), as well as the canine GHS-receptor (U.S. Pat. No. 6,645,726), and their use 3. Isocratic plateau of 10 min at 70% ACN, 30% of a 50 mM in generally identifying agonists and antagonists thereof are solution of CHCOONH in H.O. 55 4.5 min gradient to 35% ACN, 65% of a 50 mM solution of known. CHCOONH in H.O. Appropriate methods for determining the functional activ 5 . Isocratic plateau of 10 min at 35% ACN, 65% of a 50 mM ity of compounds of the present invention that interact at the solution of CHCOONH in H.O. human ghrelin receptor are also described below. 6. Flow: 0.5 mL/min 60 A. Competitive Radioligand Binding Assay (Ghrelin Recep 7. Column temperature: room temperature tor) 8. Sample temperature: room temperature The competitive binding assay at the human growth hor mone secretagogue receptor (hGHS-R1a) was carried out Method Chiral B: Grad40A-05 (column: Chiralcel OD-RH, analogously to assays described in the literature. (Bednarek 0.46 cmx15 cm): 65 M A et al. Structure-function studies on the new growth 1. Isocratic plateau of 40 min at 40% ACN, 60% of a solution hormone-releasing peptide ghrelin: minimal sequence of 50 mM of CHCOONH in H.O. ghrelin necessary for activation of growth hormone secreta US 7,491.695 B2 77 78 gogue receptor 1a; J. Med. Chem. 2000, 43, 4370-4376: of either binding buffer (total binding, N=5), 1 uM ghrelin Palucki, B. L. et al. Spiro (indoline-3,4'-piperidine) growth (non-specific binding, N=3) or the appropriate concentration hormone secretagogues as ghrelin mimetics: Bioorg. Med. of test compound (N=2 for each test concentration). The Chem. Lett. 2002, 11, 1955-1957.) reaction was initiated by addition of 40 uL of ''Il-ghrelin Materials (final conc. 0.0070-0.0085 nM) to each well. Plates were Membranes (GHS-R/HEK293) were prepared from HEK sealed with TopSeal-A, Vortexed gently and incubated at rt for 293 cells stably transfected with the human ghrelin receptor 30 min. The reaction was arrested by filtering samples (hGHS-R1a). These membranes were provided by Perki through Multiscreen Harvest plates (pre-soaked in 0.5% nElmer BioSignal (#RBHGHSM, lotil 1887) and utilized at a polyethyleneimine) using a Tomtec Harvester, washed 9 quantity of 0.71 ug/assay point. 10 times with 500 uL of cold 50 mM Tris-HCl (pH 7.4, 4° C.), 1. ''Il-Ghrelin (PerkinElmer, #NEX-388); final concentra and then plates were air-dried in a fumehood for 30 min. A tion: 0.0070-0.0085 nM bottom seal was applied to the plates prior to the addition of 2. Ghrelin (Bachem, #H-4864); final concentration: 1 uM 25uL of MicroScint-0 to each well. Plates were than sealed 3. Multiscreen Harvest plates-GF/C (Millipore, with TopSeal-A and counted for 30 sec per well on a Top #MAHFC1H60) 15 Count Microplate Scintillation and Luminescence Counter 4 . Deep-well polypropylene titer plate (Beckman Coulter, (PerkinElmer) using a count delay of 60 sec. Results were #267006) expressed as counts per minute (cpm). . TopSeal-A (PerkinElmer, #6005185) Data were analyzed by GraphPad Prism (GraphPad Soft . Bottom seal (Millipore, HMATAHOP00) ware, San Diego, Calif.) using a variable slope non-linear . MicroScint-0 (PerkinElmer, #6013.611) regression analysis. K. values were calculated using a K. . Binding Buffer: 25 mM Hepes (pH 7.4), 1 mM CaCl, 5 value of 0.01 nM for ''Il-ghrelin (previously determined mM MgCl, 2.5 mM EDTA, 0.4% BSA during membrane characterization). Assay Volumes D, values were calculated using the following formula: Competition experiments were performed in a 300 ul fil 25 tration assay format. 1. 220 uL of membranes diluted in binding buffer test concentration with maximal displacement 2. 40 uL of compound diluted in binding buffer non-specific binding Dmax = 1 - x 100 3. 40 uL of radioligand (III-Ghrelin) diluted in binding total binding- non-specific binding buffer 30 Final test concentrations (N=1) for compounds of the present invention: where total and non-specific binding represent the cpm obtained in the absence or presence of 1 LM ghrelin, respec 10, 1, 0.5,0.2, 0.1, 0.05, 0.02, 0.01, 0.005, 0.002, 0.001 uM. tively. Compound Handling 35 Compounds were provided frozen on dry ice at a stock Binding activity at the gherlin receptor for representative concentration of 10 mM diluted in 100% DMSO and Stored at compounds of the present invention is shown below in Table -80° C. until the day of testing. On the test day, compounds 3A through 3E. Compound structures for Tables 3A, 3B and were allowed to thaw at rt O/N and then diluted in assay buffer 3D are presented with the various groups as defined for the 40 general structure of formula I. For Tables 3B and 3D, in all according to the desired test concentrations. Under these con entries, m, n and pare 0; X, Z and Z are each NH. For Table ditions, the maximal final DMSO concentration in the assay 3B. R is H for all entries. The tethers (T) are illustrated with was 0.1%. the bonding to X and Z as indicated. The compounds them Assay Protocol selves are shown for Tables 3C and 3E. Competitive binding In deep-well plates, 220 uL of diluted cell membranes 45 curves for representative compounds 1, 2, 3, 4 and 25 are (final concentration: 0.71 ug/well) were combined with 40 uL shown in FIG. 4.
US 7,491.695 B2 87 88 (Wu)
s s s s s s | s s 84. X s 3888s X s N × N ×
12, , , , , , , ,
*HOO -i i r , , , , i.
H
-ULIO xpd
US 7,491.695 B2 91 92 (Wu)
s 1888.8888.888s N × N - N × N × N × N × N ×
an I k --I ......
*HOO H0 H0 H0 |------
-ULIO xpd —N99
US 7,491.695 B2 95 96
222 (, , , , , ,
IZU
H0 *HOO
I O 2 O ......
-ULIO xpd =N.TÁ =NÍZÁL
US 7,491.695 B2 99 100 (Wu)
s 1888s N × N × -N - 8888N × N ×
an I I O
V A 3. k . 22, 22.
-: : ±%
H0 H0 H0 H0 H0 H0 H0 H0 ------
-ULIO xpd =NI06
US 7,491.695 B2 107 108 (Wu) 8N × ZZ. ZZ. . . .N × .N ×
(, , , , , , ,
—N() G. G. G. G. G. G. G.
H0 H0 H0 H0 H0 H0
-ULIO xpd
US 7,491.695 B2 131 132 (Wu)
US 7,491.695 B2 133 134
TABLE 3B Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs R6 Tether KI (nM) 298 v. CH3 H CH3 Xo H Clar
F b X
299 CH3 H CH3 H OC C o1a-X
301 H H C OC. OH 1a-X
303 H H Caro1a-X
305 CH3 H CH3 H (22)? N A o1-X
306a CH3 H CH3 H N
v’s Xo F a o1a-X~ on
306b diastereomer 307 a- CH3 H CH3 Xo H \- /" F
3O8 - CH3 H CH3 Xo H OC. C O 1-X
309 CH3 H CH3 H OC F o1a-X
310 CH3 H CH3 s H OC A o1a-X
311 CH3 H CH3 H OC o1a-X OH
312 CH3 H CH3 F H OC 1-X
313 CH3 H CH3 F H OC o1a-X US 7,491.695 B2 135 136
TABLE 3B-continued
Binding Activi at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs Tether KI (nM) 314 a. CH3 H CH3 C (22)?
C
315 CH3 H CH3 XC (22)? C
316 Y CH3 H CH3 Xro (22)? F
317 a. CH3 H CH3 F (22)?
F
3.18 CH3 H CH3 F v’s F (22)? F F F 319 v. CH3 H CH3 Xo (22)? Br
320 • CH3 H CH3 Xo (22)? I
321 a. CH3 H CH3 (22)?
nNSa
322 CH3 H CH3 v’s F (22)? F F 323 v’. CH3 H CH3 Xo GH, (22)? Q CH
324 CH3 H CH3 H (22)? v’s NA
325 ~% H H (22)? US 7,491.695 B2 137 138
TABLE 3B-continued inding Activi at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R R R 7 Rs R6 KI (nM) 326 H Xro" (22)?
327a H F (22)?
F
3.27b. diastereomer 328 H Xor(H, (22)?
329 (22)?
330 (22)?
C
331a. (22)? " Xro? F
331b diastereomer 3.32a H F F (22)? F
332b diastereomer 333 H S l (22)?
335 CH3 H CH3 (22)? Y O Xo F "
336 CH3 H CH3 (22)? Xo F "
337 CH3 H CH3 H N (22)? Xo F " 338 H N-yé. CH3 H CH3 H (22)? US 7,491.695 B2 139 140
TABLE 3B-continued Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs R6 Tether KI (nM)
339 HN ~% CH3 H CH3 H OCC. (22)? F 1n-X
340 -S Y-ye CH3 H CH3 H OCC. (22)? F 1n-X
341 SA H H C Caro1-X
342 aN% H H C OCaro1a-X
343 OGH; H H C CC,X (22)? e O1a
344 Y ou, H H C OCaro1-X
345a O~&lik H H C OC.1-X
346 H H C OC.1a-X
347 H H 3.ro CC, (22)? o1a-X
348a. CH3 CH3 H H 3.no CC, (22)? o1a-X
34.8b diastereomer 353a. CH3 H CH3 H as (22)? R F
353b diastereomer 3S4 • CH3 H CH3 Xo H (22)? F o1a-X US 7,491.695 B2 141 142
TABLE 3B-continued Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R. R. R7 Rs R6 Tether KI (nM) 355 H F CH3 H CH3 Xo (22)? F O1N1 X
356 H F F CH3 H CH3 Xo F o1n X
357 CH3 H CH3 Xo H N.”S N OCo1a-X F O H 358a. H H C n122, R
358b diastereomer 359 C (22)? o1-X 360 XO"C X \- / (22)? 361 C CCs (22)?
362 C (22)? X
363 Xro" \ /" 364 Xror \- /" 365 Xor \ /" 366 XO" \ /" US 7,491.695 B2 143 144
TABLE 3B-continued Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs R6 Tether KI (nM)
367 • CH3 H CH3 Xo H Orior B
F b X 368a. ~% H H Xro" Clar B b-( X 368b diastereomer B
369 ~% H H XO" OCo1n X B
370 H H C X (22)? C
371 ~% H H C CO (22)? B
372 V 7 CH3 H CH3 Xo H CCX A. F F o1a1
373 CH3 H CH3 Xo H "ocal B V7 F O1a X
374 CH3 H CH3 H O B W, v’s F 2, OC,1-X
375 CH3 H CH3 H C v’s (22)? F o1n X
376 CH3 H CH3 Xo H Orror C v F O X
377 CH3 H CH3 H O O C
v’s Xo F rors
378 CH3 H CH3 H C v’s (22)? F o1n X
379 CH3 H CH3 H B v’s (22)? F o1n X US 7,491.695 B2 145 146
TABLE 3B-continued Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs R6 Tether KI (nM) 380 ~% C H H Xor h(22)? o1a-X 381 ~% C H H XO" o1a-X(22)?
1-X
383 ~% H H XO" F OC1n-X
384 ~% H H Xror "ocalo1-X
385 ~% H H Xor Nsa o1a-X(22)?
386 H H C
o1a-X 387 ~% C H H Xor c.(22)? 388 H H C o1-X
389a v. CH3 H CH3 Xo H (22)? F c. US 7,491.695 B2 147 148
TABLE 3B-continued
Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R R Rs R6 Tether KI (nM)
389b diastereomer 390 H C Xa-1a-N1-22
391 CH3 H CH3 F H (22)? F OC F 392 Or (22)?
393
394 CH3 H CH3 (22)? OCC.or 395 OC.(22)?
398 CH3 H CH3 X \- /"(22)
399a (22)? C O
399b diastereomer 400 CH3 H CH3 (22)? B
401 CH3 H CH3 US 7,491.695 B2 149 150
TABLE 3B-continued Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention
Com pound R2 R R R7 Rs R6 Tether KI (nM)
402a H H C Car B O1a- X
402b diastereomer B Binding activity determined using standard method, expressed as follows: A = 0.1-10 nM; B = 10-100 nM; C = 0.1-1.0 M
TABLE 3C 2O TABLE 3C-continued
Binding Activity at the Human Ghrelin Receptor for Representative Binding Activity at the Human Ghrelin Receptor for Representative Compounds of the Invention Compounds of the Invention 25 Com- Com pound Structure Ki (nM) pound Structure Ki (nM)
18 B 349 C B O O O O NH HN NH HN O 35 O "> HN "> HN O O o 40
45
50 334 C B 350 C C O O O y- O y NH HN 55 NH HN O O HN HN
60 a O & O o
65