US 2011 02881 63A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0288163 A1 Fraser et al. (43) Pub. Date: Nov. 24, 2011 (54) METHODS OF USING MACROCYCLC 60/622,005, filed on Oct. 27, 2004, provisional appli MODULATORS OF THE GHRELIN cation No. 60/642,271, filed on Jan. 7, 2005. RECEPTOR Publication Classification (76) Inventors: Graeme L. Fraser, Quebec (CA); (51) Int. Cl. Hamid R. Hoveyda, Quebec (CA); A 6LX 3L/395 (2006.01) Mark L. Peterson, Quebec (CA) A6IP 43/00 (2006.01) (21) Appl. No.: 13/173,929 (52) U.S. Cl. ........................................................ S14/450 (57) ABSTRACT (22) Filed: Jun. 30, 2011 The present invention provides novel conformationally-de fined macrocyclic compounds that have been demonstrated to Related U.S. Application Data be selective modulators of the ghrelin receptor (growth hor (60) Division of application No. 12/333,026, filed on Dec. mone secretagogue receptor, GHS-R1a and Subtypes, iso 11, 2008, which is a continuation of application No. forms and variants thereof). Methods of synthesizing the 1 1/149,512, filed on Jun. 10, 2005, now Pat. No. 7,491, novel compounds are also described herein. These com 695, which is a continuation-in-part of application No. pounds are useful as agonists of the ghrelin receptor and as 10/872,142, filed on Jun. 18, 2004, now Pat. No. 7,521, medicaments for treatment and prevention of a range of medi 420. cal conditions including, but not limited to, metabolic and/or endocrine disorders, gastrointestinal disorders, cardiovascu (60) Provisional application No. 60/479,223, filed on Jun. lar disorders, obesity and obesity-associated disorders, cen 18, 2003, provisional application No. 60/621,642. tral nervous system disorders, genetic disorders, hyperprolif filed on Oct. 26, 2004, provisional application No. erative disorders and inflammatory disorders. Patent Application Publication Nov. 24, 2011 Sheet 1 of 21 US 2011/02881 63 A1 dnou66u?33.30.Id== |EH[150]- Z -*-faa-i-O€gH-O?4)-------- E Patent Application Publication Nov. 24, 2011 Sheet 2 of 21 US 2011/02881 63 A1 z914 O Patent Application Publication Nov. 24, 2011 Sheet 3 of 21 US 2011/02881 63 A1 (º1)uaqyaife?ued Patent Application Publication Nov. 24, 2011 Sheet 4 of 21 US 2011/02881 63 A1 Target; h(GHS-R Radioligand; 125Ghrelin 50 i Compound 1 FIGURE 4 Log M of Compound 1 Target; h(GHS-R 1 Radioligand; 125Ghrelin 75 * . it: '... Compound 2 -9 -8 -7 -6 -5 LOg M of Compound 2 FIGURE 4B Patent Application Publication Nov. 24, 2011 Sheet 5 of 21 US 2011/02881 63 A1 Target; h(GHS-R Radioligand; 125Ghrelin 7 5 i s Compound 3 -9 -8 -7 -6 5 LOg M of Compound FIGURE 4C Target; hCGHS-R Radioligand: 125Ghrelin 1, g 75- \s 50- \ N isf 25. Y. Compound 4 O- K= n13t3nM = 6 is . -9 -8 -, -6 -5 LogM of Compound 4 r FIGURE 4D Patent Application Publication Nov. 24, 2011 Sheet 6 of 21 US 2011/02881 63 A1 Target; he hS-R Radioligand; 125Ghrelin f Ki = 0.34 nM Compound 25 LOg M of compound s FIGURE 4E Compound 1 -10 -9 -8 -7 -6 -5 -4 Log Concentration (M) FIGURE 5A Patent Application Publication Nov. 24, 2011 Sheet 7 of 21 US 2011/02881 63 A1 Compound 2 125 OO 7 5 50 0 - FIGURE 5B Log Concentration (M) Compound 3 100 75 25 i5. o 34 I -- O5O . -10 -9 -8 .7 -6 -5 -4 FIGURE 5C 09 Concentration (M) Patent Application Publication Nov. 24, 2011 Sheet 8 of 21 US 2011/02881 63 A1 Compound 4 OO ul vu g 5 O -1 25- s: EC50=16.6 nM Hill slope = 0.8 O- g -10 9 -8 -7 -6 -5 -4 FIGURE 5D Log Concentration (M) Compound 5 OO T 57 O5 l 2 5 EC50=298nM Hill slope = 1.5 FIGU RE st -9 -8 Log Concentration-7 (M)-6 -5 -4 Patent Application Publication Nov. 24, 2011 Sheet 9 of 21 US 2011/02881 63 A1 s O 6 O 120 180 24 300 360 FIGURE 6A Time (min) O 3O 6D 90 2 15 SO 20 2.O 270 300 33 3 SO 390 FIGURE 6B Tirne (min) Patent Application Publication Nov. 24, 2011 Sheet 10 of 21 US 2011/02881 63 A1 15 30 A5 SO 75 90 105 12O 135 5. FIGURE 6C Tirne min) SOO 1AOO 12DD 1OOO BOO GOO 4OO 2O) O 15 3D 45 SO 75 90 105 120 35 15 FIGURE 6D Time (nir) Patent Application Publication Nov. 24, 2011 Sheet 11 of 21 US 2011/02881 63 A1 OB E 0.6 8 d: c S. D.4 E E S 2 0.2- () 'ra O O ; : s 30 100 300 g g Compound 25 3 p 8 Clgkg) 38 FIGURE 7A : D.8 Ovehicle 6 metoclopramide (10 mg/kg) g O. 4. SaGHRP-6 (20 pg/kg) ESCompound 298 (100 pg/kg) O. 2 , 3 d 3. O 3. S - FIGURE 7B ( ) Patent Application Publication Nov. 24, 2011 Sheet 12 of 21 US 2011/02881 63 A1 B Onaive S O is PO + Wehicle so POI + Compound 298 2d O ST 10 20 30 40 50 60 70 BO Georetic Center of G raisit Naive - 35 cm PO - S Cl Compound 298 - 16 cm FIGURE 8 Patent Application Publication Nov. 24, 2011 Sheet 13 of 21 US 2011/02881 63 A1 3DO Vehicle - 0.3 ml A. 225 150 75 300 225 150 75 3OO 225 150 75 300 ' ' Ghrelin. 5ug D 225 i 150 75 OOO 1100 1200 1300 400 1500 1600 Time (hours) FIG. 9 Patent Application Publication Nov. 24, 2011 Sheet 14 of 21 US 2011/02881 63 A1 Compound 298 Binding to h(GHS-R1a 100- y y N W o, 75 y \ 2 50 y is 25 Ki = 15 nM Dmax 96% N Hill slope = 0.87 FIGU RE io -10 Log Concentration-8 (M) -6 -4 100 v Compound 298 50 EC50 = 25 nM Oghrelin EC50 = 2nM FIGU RE 1. L 10 Compound (M)-8 -6 Patent Application Publication Nov. 24, 2011 Sheet 15 of 21 US 2011/02881 63 A1 Ghelin (5ug) O Grein (5ug) + Compound 298 (300 uglkg) Compound 298 Ghrein Time (hours) FIGURE 12 Patent Application Publication Nov. 24, 2011 Sheet 16 of 21 US 2011/02881 63 A1 - 100 Blank 2 10-9M , 10-8M 50 ... 107M t -6- . 105M FIGURE 13B to 3 is ; ; ; 150 DeSensitization h Concentration response Curve a Ghrein a GHRP-6 E 100 v Capromorelin s Compound 298 50 e Compound 430 is 0 r 5 O -11 -10 -9 8 -7 -6 -5 FIGURE 13C Pretreatment concentration (LOg M) Patent Application Publication Nov. 24, 2011 Sheet 17 of 21 US 2011/02881 63 A1 1.00 O,75 0.50 0.25 0.00 5 2 O 50 5 20 50 ghrelin GHRP-6 FIGURE 14A (ug/kg) (ug/kg) 1.00 0.75 50 10 30 100 300 Compound 298 FIGURE 14B : Patent Application Publication Nov. 24, 2011 Sheet 18 of 21 US 2011/02881 63 A1 8 D. a We E. P O - We icia ised PO - Compound 298 S Geo effic Ce per of G trays if Naive - 35 c in P of 6 cri Compound 298 - 16 cm FIGURE 15 Patent Application Publication Nov. 24, 2011 Sheet 19 of 21 US 2011/02881 63 A1 FIG. 1.6A morphine 3 mg/kg (s.c.) 57 O5 2 5 naive Vehicle O.3 1 TZP-101 mg/kg (i.v.) FIG. 16B morphine 3 mg/kg (S.C.) naive vehicle 0.3 1 TZP-101 mg/kg (i.V.) Patent Application Publication Nov. 24, 2011 Sheet 20 of 21 US 2011/02881 63 A1 C C O. E O U r G s g=u uepeu Ouluoo JueMOS JO % u pueuoo toeuoS Patent Application Publication Nov. 24, 2011 Sheet 21 of 21 US 2011/02881 63 A1 o O O E O U wn G) wa (0=u :OO }uenos) G-v= u uleeu Oluod uemos go 9% u pueu oo Loeuos US 2011/02881 63 A1 Nov. 24, 2011 METHODS OF USING MACROCYCLC and skeletal muscles. This receptor was identified and cloned MODULATORS OF THE GHRELIN prior to the isolation and characterization of the endogenous RECEPTOR peptide ligand and is distinct from other receptors involved in the regulation of growth hormone (GH) secretion, in particu RELATED APPLICATION INFORMATION lar, the growth hormone-releasing hormone (GHRH) recep 0001. This application is a continuation application of tOr. U.S. patent application Ser. No. 1 1/149.512, filed Jun. 10, 0004. A unique characteristic of both the rat and human 2005, which is a continuation-in-part under 35 U.S.C. S 120 of peptides is the presence of the n-octanoyl (Oct) moiety on U.S. patent application Ser. No. 10/872,142, filed Jun. 18. Ser. However, the des-acyl form predominates in circula 2004, currently pending, which claims the benefit under 35 tion, with approximately 90% of the hormone in this form. U.S.C. S119(e) of U.S. Provisional Patent Application Ser. This group is derived from a post-translational modification No. 60/479,223, filed Jun. 18, 2003. The continuation-in-part and appears relevant for bioactivity and possibly also for application also claims the benefit under 35 U.S.C. S 119(e) of transport into the CNS. (Banks, W.A.: Tschöp, M.: Robinson, U.S. Provisional Patent Application Ser. No.